Compounds and oligonucleotides derived from N-acetylgalactosamine (GalNAc)

Abstract

To provide compounds and oligonucleotides derived from N-acetylgalactosamine (GalNAc). [Solution] Compounds derived from N-acetylgalactosamine (GalNAc), modified oligonucleotides, methods for modulating protein function, and methods for treating diseases, disorders, and symptoms in subjects are provided herein. The present invention relates to compounds, for example any of those described herein, modified oligonucleotides, methods for modulating protein function, and methods for targeting diseases, disorders, and symptoms in subjects. These methods may include the compounds and modified oligonucleotides disclosed herein.

Description

[Technical Field]

[0001] Related applications This application claims priority to U.S. Provisional Application No. 63 / 251,580 filed on 1 October 2021, U.S. Provisional Application No. 63 / 141,300 filed on 25 January 2021, U.S. Provisional Application No. 63 / 131,317 filed on 29 December 2020, and U.S. Provisional Application No. 63 / 089,936 filed on 9 October 2020, as well as U.S. Provisional Application No. 63 / 146,276 filed on 5 February 2021, and U.S. Provisional Application No. 63 / 113,801 filed on 13 November 2020. Each of these U.S. Provisional Applications is incorporated herein by reference. [Background technology]

[0002] background When using compounds for therapeutic, preventive, or diagnostic purposes, it is often desirable that these compounds be delivered to specific locations (e.g., desired cells) to enhance therapeutic or preventive effects, or to be advantageous for diagnostic purposes. This is frequently the case when attempting to deliver therapeutic compounds in vivo. Furthermore, the efficient delivery of compounds to specific locations can limit or potentially eliminate unintended consequences (e.g., off-target effects) that may be caused by the administration of the compound. One strategy to facilitate the in vivo delivery of compounds, such as therapeutic, preventive, or diagnostic compounds, to desired locations is to bind or attach the compound to a targeting ligand.

[0003] One class of compounds that can be targeted using targeting ligands are oligomeric compounds. Oligomers containing nucleotide sequences that are at least partially complementary to a target nucleic acid have been shown to alter the function and activity of their target both in vitro and in vivo. When delivered to cells containing a target nucleic acid (e.g., mRNA), oligomeric compounds have been shown to modulate the expression of this target, resulting in altered transcription or translation of this target nucleic acid. In certain examples, these oligomeric compounds may reduce the expression of their genes by inhibiting the nucleic acid target and / or inducing the degradation of this target nucleic acid.

[0004] When the target nucleic acid is mRNA, one mechanism by which an oligomeric compound that inhibits expression can regulate the expression of this mRNA target is via RNA interference. RNA interference is a biological process in which RNA or RNA-like molecules (e.g., chemically modified RNA molecules) can silence gene expression through degradation. Furthermore, RNA-like molecules, which may also include single-stranded RNA and modified nucleotides and may have one or more non-phosphodiester bonds, can also alter the expression of target nucleic acids, such as target mRNA.

[0005] Another class of compounds that can be targeted using targeting ligands are small molecule compounds. Small molecule compounds (e.g., organic compounds with a molecular weight of approximately 1000 daltons or less) have been shown, when localized to their target, typically alter the function and / or activity of that target, resulting in the modulation or improvement of disease and / or disease symptoms, or typically, their usefulness as diagnostic markers. By delivering the compound more efficiently to a specific site, undesirable consequences that may be caused by the administration of this compound (e.g., off-target effects) can be limited or potentially eliminated. This can be done, and it can provide improved localization of diagnostic compounds. [Overview of the project] [Means for solving the problem]

[0006] Brief summary of the invention The present invention relates to compounds (for example, any of those described herein), modified oligonucleotides, methods for modulating protein function, and methods for targeting diseases, disorders, and symptoms in subjects. These methods may include the compounds and modified oligonucleotides disclosed herein.

[0007] It will be understood that the embodiments of the present invention discussed below with respect to preferred variable options may be employed individually or in combination with one or more embodiments or preferred variable options of the present invention, as each combination is expressly enumerated herein.

[0008] In one aspect, compounds of formulas (Ia), (Ib), (Ic), and (Id), as well as their salts, solvates, and hydrates: [ka] The following is provided, in formulas (Ia), (Ib), (Ic), and (Id): [ka] is ring A, where each ring A is independently a substituted carbocyclyl or a substituted heterocyclyl as needed; [ka] is ring B, where each ring B is independently an optionally substituted aryl or optionally substituted heteroaryl; Each n is independently 0, 1, 2, 3, or 4; Each Y is independently O, CH2, S, S(=O), S(=O)2, NH, substituted amino, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Z independently has an optionally substituted alkyl group, optionally substituted alke A nyl group, optionally substituted alkynyl groups, or ethylene glycol; Each R1 is independently an alkyl-O-phosphoramidite, an alkylphosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite; R2 and R5 are, independently, optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; R3 and R6 are independently, as needed, alkyl-O-GalNAc, as needed, alkenyl-O-GalNAc, as needed, alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; and Each R4 is independently alkyl-O-GalNAc as needed, alkenyl-O-GalNAc as needed, alkynyl-O-GalNAc as needed, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc.

[0009] In certain embodiments, this compound is a compound of formula (Ia).

[0010] In certain embodiments, this compound is a compound of formula (Ib).

[0011] In certain embodiments, this compound is a compound of formula (Ic).

[0012] In certain embodiments, this compound is a compound of formula (Id).

[0013] In certain embodiments, R2 and R3 are the same.

[0014] In certain embodiments, ring A is a substituted carbocyclyl as needed. In certain embodiments, ring A is a substituted heterocyclyl as needed.

[0015] In certain embodiments, ring B is an optionally substituted aryl. In certain embodiments, ring B is an optionally substituted heteroaryl.

[0016] In another context, compounds of formulas (II-a), (II-b), and (II-c), as well as their salts, solvates, and hydrates: [ka] The following are provided herein, in formulas (II-a), (II-b), and (II-c): Each n is independently 0, 1, 2, 3, or 4; Each Y is independently O, CH2, S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Z is independently an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, or ethylene glycol; Each R1 is independently an alkyl-O-phosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite; R5, R6, R7, and R8 are each independently of optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; and R9 is H, adenine, guanine, thymine, cytosine, uracil, inosine (I), or a nucleic acid base variant.

[0017] In certain embodiments, this compound is the compound of formula (II-a).

[0018] In certain embodiments, this compound is the compound of formula (II-b).

[0019] In certain embodiments, this compound is a compound of formula (II-c).

[0020] In certain embodiments, three of R5, R6, R7, and R8 are each independently optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, and optionally substituted The substituted R5 is alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc, and the remaining one R5, R6, R7, or R8 is independently H, alkyl, alkenyl, alkynyl, halogen, substituted amine, thiol, or amide. In certain embodiments, two of R5, R6, R7, and R8 are each the same. In certain embodiments, three of R5, R6, R7, and R8 are each the same, and the remaining R5, R6, R7, or R8 is independently H, alkyl, alkenyl, alkynyl, halogen, substituted amine, thiol, or amide.

[0021] In certain embodiments, R9 is H. In certain embodiments, R9 is adenine, guanine, thymine, cytosine, or uracil.

[0022] In another context, compounds of formulas (III-a) and (III-b), as well as their salts, solvates, and hydrates: [ka] The following are provided herein, in formulas (III-a) and (III-b): Each R1 is independently an alkyl-O-phosphoramidite, an alkylphosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite; Each R2 is independently an optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; Each R3 independently has the following options: optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O -It is alkyl-GalNAc; and Each R4 is independently optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc; alkyl-S(=O)₂-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)₂-NH-alkyl-GalNAc, alkyl-PP(=O)(-O⁻)-NH-alkyl-GalNAc, alkyl-O-P(=O)(-O⁻)-O-alkyl-GalNAc, alkyl-O-P(-O⁻)(=S)-O-alkyl-GalNAc, or alkyl-O-P(-S⁻)(=S)-O-alkyl-GalNAc.

[0023] In a specific embodiment, this compound is a compound of formula (III-a).

[0024] In a specific embodiment, this compound is a compound of formula (III-b).

[0025] In another aspect, a compound of the formula: ##STR1## ##STR2## , and salts, solvates, and hydrates thereof are provided herein, wherein: each n is independently an integer of 0 to 10, inclusive of 0 and 10; each m is independently an integer of 0 to 10, inclusive of 0 and 10; and the base is adenosine, uracil, thymine, cytosine, inosine, or guanosine.

[0026] In another aspect, provided herein are compounds of formulas (IV-a), (IV-b), (IV-c), and (IV-d): ##STR3## Provided herein is the compound, as well as salts, solvates and hydrates thereof, In formulas (IV-a), (IV-b), (IV-c), and (IV-d): n1 is independently an integer from 1 to 10, inclusive of 1 and 10; n2 is independently an integer from 1 to 10, inclusive of 1 and 10; n3 is independently an integer from 1 to 10, inclusive of 1 and 10; n4 is independently an integer from 1 to 10, inclusive of 1 and 10; each X is independently H, alkyl-GalNAc, or PEG-GalNAc; and each Y is independently H, alkyl-GalNAc, or PEG-GalNAc.

[0027] In one aspect, modified oligonucleotides comprising any of the compounds disclosed herein are provided herein. In certain embodiments, the modified oligonucleotide comprises an siRNA, miRNA, ADAR mobilizing molecule, ADAR targeting molecule, guide RNA, or antisense nucleic acid. In some embodiments, the modified oligonucleotide comprises any of the compounds disclosed herein, wherein (i) the heterocyclyl comprises a heteroatom that is oxygen, (ii) the heterocyclyl comprises at least one heteroatom that is sulfur, or (iii) the heterocyclyl does not contain a nitrogen heteroatom. In some embodiments, the modified oligonucleotide comprises any of the compounds disclosed herein, wherein ring A is not cyclopentyl, pyrrolidinyl, piperidinyl, or morpholinyl. In some embodiments, the modified oligonucleotide comprises any of the compounds disclosed herein, wherein (i) the heteroaryl does not contain a nitrogen heteroatom, (ii) the heteroaryl comprises a heteroatom that is oxygen, or (iii) the heteroaryl comprises a heteroatom that is sulfur. In some embodiments, the modified oligonucleotide comprises any of the compounds disclosed herein, wherein ring A is not phenyl, pyridinyl, 1,3-pyrimidinyl, 1,4-pyrimidinyl, 1-quinolinyl, or 9H-prinyl.

[0028] In another context, compounds of formula (V), as well as their salts, solvates, and hydrates: [ka] The following is provided herein, in formula (V): R 1 These are H, adenine, guanine, thymine, cytosine, or uracil; R 2 H, protecting group (PG), [ka] and; L1 is alkyl or alkyl-C(=O)-NH-alkyl; L 2 is alkyl or alkyl-C(=O)-NH-alkyl; R 3 is H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 is H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 is -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 6 is -C=(O)-NH-(CH2CH2O) n -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; j is an integer from 1 to 10, inclusive of 1 and 10; k is an integer from 1 to 10, inclusive of 1 and 10; m is an integer from 1 to 10, inclusive of 1 and 10; and n is an integer from 1 to 10, inclusive of 1 and 10.

[0029] In another aspect, provided herein is a compound of formula (V-a), and salts, solvates, and hydrates thereof:

Chemical Formula

[0030] In another aspect, provided herein is a compound of formula (V-b), and a salt, solvate, and hydrate thereof:

Chemical Formula

[0031] In a particular embodiment, R 3 , R 4 , R 5 , and R 6 They are the same. In a particular embodiment, R 3 , R 5 , and R 6 They are the same. In a particular embodiment, R 3 or R 4 H is H.

[0032] In a particular embodiment, L 1 and L 2 They are the same.

[0033] In a particular embodiment, L 1 and L 2 Each is independently an alkyl group; R 3 H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k-GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; and R 6 -C=(O)-NH-(CH2CH2O) n It is either -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc.

[0034] In a particular embodiment, L 1 and L 2 Each of them is independently alkyl-C(=O)-NH-alkyl; R 3 H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; and R 6 -C=(O)-NH-(CH2CH2O) n It is either -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc.

[0035] In a particular embodiment, R 4 H is H. In a particular embodiment, L 1 and L 2 Each of them is independently an alkyl group; R 3 -C=(O)-NH-(CH2CH2O) jis -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 is H; R 5 is -C(=O)-NH-(CH2CH2O) m -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; and R 6 is -C(=O)-NH-(CH2CH2O) n -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc .

[0036] In certain embodiments, L 1 and L 2 are each independently alkyl-C(=O)-NH-alkyl; R 3 is -C(=O)-NH-(CH2CH2O) j -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 is H; R 5 is -C(=O)-NH-(CH2CH2O) m -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; and R 6 is -C(=O)-NH-(CH2CH2O) n -GalNAc or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc.

[0037] In certain embodiments, R 3 is -C(=O)-NH-(CH2CH2O) j -GalNAc; R 4 is H; R 5 is -C(=O)-NH-(CH2CH2O) m -GalNAc; and R 6 is -C(=O)-NH-(CH2CH2O) n -GalNAc.

[0038] In a particular embodiment, R 3 This is -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H is R 5 is -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; and R 6 It is -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc.

[0039] In certain embodiments, j, m, and n are each independently integers from 4 to 10, including 4 and 10. In certain embodiments, j, m, and n are each the same integers from 4 to 10, including 4 and 10.

[0040] In a particular embodiment, R 1 is H. In a particular embodiment, R 1 is adenine, guanine, thymine, cytosine, or uracil. In certain embodiments, R 1 is adenine. In certain embodiments, R 1 is guanine. In certain embodiments, R 1 is thymine. In a particular embodiment, R 1 is cytosine. In certain embodiments, R 1 This is Uracil.

[0041] In a particular embodiment, R 2 is H. In a particular embodiment, R 2 is a protecting group (PG). In certain embodiments, PG is an oxygen protecting group. In certain embodiments, R 2 teeth, [ka] That is the case.

[0042] In certain embodiments, any of the compounds disclosed herein, or their salts, solvates, or hydrates, may be used as reagents in chemical reactions.

[0043] In another context, compounds of formula (VI), as well as their salts, solvates, and hydrates: [ka] The following is provided herein, in formula (VI): R 1 These are H, adenine, guanine, thymine, cytosine, or uracil; R 2 This is an oligonucleotide sequence; L 1 is alkyl, or alkyl-C(=O)-NH-alkyl; L 2 is alkyl, or alkyl-C(=O)-NH-alkyl; R 3 H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 6 -C=(O)-NH-(CH2CH2O) n -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; j is an integer from 1 to 10, including 1 and 10; k is an integer from 1 to 10, including 1 and 10; m is an integer from 1 to 10, including 1 and 10; and n is an integer from 1 to 10, including 1 and 10.

[0044] In another context, modified oligonucleotides comprising compounds of formula VI are provided herein. In certain embodiments, these modified oligonucleotides include siRNA, miRNA, ADAR mobilization molecules, ADAR targeting molecules, guide RNA, or antisense nucleic acids.

[0045] In one aspect, the part of formula (VII), or its salt, solvate, or hydrate: [ka] Oligonucleotides comprising the following are provided herein, in formula (VII): R 1 These are H, adenine, guanine, thymine, cytosine, or uracil; R 2 is a combination; L 1 is alkyl, or alkyl-C(=O)-NH-alkyl; L 2 is alkyl, or alkyl-C(=O)-NH-alkyl; R 3 H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R6 -C=(O)-NH-(CH2CH2O) n -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; j is an integer from 1 to 10, including 1 and 10; k is an integer from 1 to 10, including 1 and 10; m is an integer from 1 to 10, including 1 and 10; and n is an integer from 1 to 10, including 1 and 10.

[0046] In certain embodiments, this oligonucleotide comprises siRNA, miRNA, ADAR mobilization molecule, ADAR targeting molecule, guide RNA, or antisense nucleic acid.

[0047] In one aspect, compositions containing any of the compounds disclosed herein and a pharmaceutically acceptable carrier are provided herein.

[0048] In another context, a method for modulating protein function in a subject is provided herein, which involves administering one of the compounds disclosed herein to the subject.

[0049] In another context, methods for treating or improving a disease, disorder, or symptoms thereof in a subject are provided herein, and these methods include administering one of the compounds disclosed herein to the subject.

[0050] In another context, compounds of formulas (Ia), (Ib), (Ic), and (Id), as well as their salts, solvates, and hydrates: [ka] The following are provided herein, in formulas (Ia), (Ib), (Ic), and (Id): [ka] is ring A, where each ring A is independently a substituted carbocyclyl or a substituted heterocyclyl as needed; [ka] is a ring B, where each B is independently a heteroaryl substituted as needed; Each n is independently 0, 1, 2, 3, or 4; Each Y is independently O, CH2, S, S(=O), S(=O)2, NH, substituted amino, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Z is independently an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, or ethylene glycol; Each R1 is independently an alkyl-O-phosphoramidite, an alkylphosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite; R2 and R5 are, independently, optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; R3 and R6 are independently, as needed, alkyl-O-GalNAc, as needed, alkenyl-O-GalNAc, as needed, alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; and Each R4 is independently alkyl-O-GalNAc as needed, alkenyl-O-GalNAc as needed, alkynyl-O-GalNAc as needed, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc.

[0051] In another context, equations (IV-a) and (IV-d): [ka] Compounds of the same, as well as their salts, solvates, and hydrates, are provided herein. In equations (IV-a) and (IV-d): n1 is an integer from 1 to 10, including 1 and 10 independently; n2 is an integer from 1 to 10, including 1 and 10 independently; n3 is an integer from 1 to 10, including 1 and 10 independently; n4 are independent integers from 1 to 10, including 1 and 10; Each X is independently H, alkyl-GalNAc, or PEG-GalNAc; and Each Y is independently H, alkyl-GalNAc, or PEG-GalNAc.

[0052] In another context, compounds of formula (VIII), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (VIII): R 1 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 H, protecting group (PG), [ka] and; Z 1 teeth, [ka] and; Z 2 teeth, [ka] and; L 1 is alkyl, or alkyl-C(=O)-NH-alkyl; L 2 is alkyl, or alkyl-C(=O)-NH-alkyl; R 3 H, -C=(O)-NH-(CH2CH2O) j-GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 6 -C=(O)-NH-(CH2CH2O) n -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; j is an integer from 1 to 10, including 1 and 10; k is an integer from 1 to 10, including 1 and 10; m is an integer from 1 to 10, including 1 and 10; and n is an integer from 1 to 10, including 1 and 10.

[0053] In another context, compounds of formula (VIII-a), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (VIII-a): R 1 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, or nucleic acid base isocouple; R 2 H, OH, O-protecting group (PG), [ka] , linker, azide, carboxylic acid, amine, or phosphate; X1 is O, NH, CH2, or CH2O as needed; Z 1 teeth, [ka] and; Z 2 teeth, [ka] and; L 1 is alkyl, or alkyl-C(=O)-NH-alkyl; L 2 is alkyl, or alkyl-C(=O)-NH-alkyl; R 3 H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 6 -C=(O)-NH-(CH2CH2O) n -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; j is an integer from 1 to 10, including 1 and 10; k is an integer from 1 to 10, including 1 and 10; m is an integer from 1 to 10, including 1 and 10; and n is an integer from 1 to 10, including 1 and 10.

[0054] In another context, compounds of formula (VIII-b), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (VIII-b): R 1 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, or nucleic acid base isocouple; R 2 H, OH, O-protecting group (PG), [ka] , linker, azide, carboxylic acid, amine, or phosphate; X1 is O, NH, CH2, or CH2O as needed; X2 is O, NH, CH2, or CH2O as needed; Z 1 teeth, [ka] and; Z 2 teeth, [ka] and; L 1 is alkyl, or alkyl-C(=O)-NH-alkyl; L 2is alkyl, or alkyl-C(=O)-NH-alkyl; R 3 H, -C=(O)-NH-(CH2CH2O) j -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 4 H, -C=(O)-NH-(CH2CH2O) k -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 5 -C=(O)-NH-(CH2CH2O) m -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; R 6 -C=(O)-NH-(CH2CH2O) n -GalNAc, or -C(=O)-NH-alkyl-NH-C(=O)-alkyl-O-GalNAc; j is an integer from 1 to 10, including 1 and 10; k is an integer from 1 to 10, including 1 and 10; m is an integer from 1 to 10, including 1 and 10; and n is an integer from 1 to 10, including 1 and 10.

[0055] In another context, compounds of formula (IX), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (IX): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2is H, a protecting group (PG), or [ka] and; Y1 is O; Y2 is O; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0056] In some embodiments, this compound is a compound of formula (IX-a), or a salt, solvate, or hydrate thereof: [ka] And in equation (IX-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 H, OH, O-protecting group (PG), [ka] , linker, azide, carboxylic acid, or amine; Y1 is O, NH, CH2, or CH2O as needed; Y2 is O, NH, CH2, or CH2O as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0057] In another context, compounds of formula (X), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (X): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is H, a protecting group (PG), or [ka] and; Y1 is O; Y2 is O; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0058] In some embodiments, this compound is a compound of formula (Xa), or a salt, solvate, or hydrate thereof: [ka] And in equation (Xa): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 H, OH, O-protecting group (PG), [ka]

[0059] , linker, azide, carboxylic acid, or amine; Y1 is O, NH, CH2, or CH2O as needed; Y2 is O, NH, CH2, or CH2O as needed; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0060] In some embodiments, R 9 is uracil. In some embodiments, R 9 is a uracil containing a protecting group. In certain embodiments, R 9 is uracil containing a benzoyl protecting group. In some embodiments, R 9 is cytosine. In some embodiments, R 9 is a cytosine containing a protecting group. In certain embodiments, R9 R is a cytosine containing an acyl protecting group. In certain embodiments, R 9 R is a cytosine containing a benzoyl protecting group. In some embodiments, R 9 is adenine. In some embodiments, R 9 is an adenine containing a protecting group. In certain embodiments, R 9 is an adenine containing a benzoyl protecting group. In some embodiments, R 9 is guanine. In some embodiments, R 9 is a guanine containing a protecting group. In some embodiments, R 9 It contains a dimethylacetate protecting group. It is guanine.

[0061] In some embodiments, R 2 is an O-protecting group (PG). In certain embodiments, R 2 teeth, [ka] That is the case.

[0062] In some embodiments, Y1 is O. In some embodiments, Y2 is O. In certain embodiments, both Y1 and Y2 are O. In some embodiments, Y3 is CO. In some embodiments, Y4 is CO. In certain embodiments, both Y3 and Y4 are CO. In certain embodiments, Y1 and Y2 are O, and Y3 and Y4 are CO.

[0063] In some embodiments, n3 is 1. In some embodiments, n3 is 3. In some embodiments, n3 is 5. In some embodiments, n3 is 7. In some embodiments, n4 is 1. In some embodiments, n4 is 3. In some embodiments, n4 is 5. In some embodiments, n4 is 7. In some embodiments, n5 is 1. In some embodiments, n5 is 3. In some embodiments, n5 is 5. In some embodiments, n5 is 7. In certain embodiments, two of n3, n4, and n5 are 1. In certain embodiments, all three of n3, n4, and n5 are 1.

[0064] In some embodiments, this compound is of formula (IX-b): [ka] The compound is such that, in formula (IX-b), R9, R2, Y1, Y2, Y3, Y4, n1, and n2 are as defined herein.

[0065] In some embodiments, this compound is of formula (Xb): [ka] The compound is such that, in formula (Xb), R9, R2, Y1, Y2, n1, and n2 are as defined herein.

[0066] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, And this compound has the formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0067] In another context, this disclosure relates to a compound of formula (XXIX), or a salt, solvate, or hydrate thereof: [ka] Provides, in formula (XXIX): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 H, OH, O-protecting group (PG), [ka] , linker, azide, carboxylic acid, or amine; Y1 is O, NH, CH2, or CH2O as needed; Y2 is O, NH, CH2, or CH2O as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0068] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0069] In another context, compounds of formula (XI), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XI): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 This is an oligonucleotide sequence; Y1 is O; Y2 is O; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0070] In another context, compounds of formula (XI-a), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XI-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is an oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0071] In another context, compounds of formula (XII), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XII): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 This is an oligonucleotide sequence; Y1 is O; Y2 is O; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0072] In another context, compounds of formula (XII-a), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XII-a): R 9H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is an oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0073] In another context, oligonucleotides comprising the part of formula (XIII), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XIII): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is a combination; Y1 is O; Y2 is O; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0074] In another context, oligonucleotides comprising the part of formula (XIII-a), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XIII-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This involves binding to the oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0075] In another context, oligonucleotides comprising the part of formula (XIV), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XIV): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is a combination; Y1 is O; Y2 is O; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0076] In another context, oligonucleotides comprising the part of formula (XIV-a), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XIV-a): R 9H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This involves binding to the oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0077] In another context, compounds of formula (XV), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XV): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 This is an oligonucleotide sequence; Y1 is O; Y2 is O; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0078] In another context, compounds of formula (XV-a), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XV-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is an oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0079] In another context, compounds of formula (XVI), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XVI): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is an oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0080] In another context, compounds of formula (XVI-a), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XVI-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is an oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0081] In some embodiments, R 9 is uracil. In some embodiments, R 9 is cytosine. In some embodiments, R 9 is adenine. In some embodiments, R 9 It is guanine.

[0082] In some embodiments, Y1 is O. In some embodiments, Y2 is O. In certain embodiments, both Y1 and Y2 are O. In some embodiments, Y3 is CO. In some embodiments, Y4 is CO. In certain embodiments, both Y3 and Y4 are CO. In certain embodiments, Y1 and Y2 are O, and Y3 and Y4 are CO.

[0083] In some embodiments, n3 is 1. In some embodiments, n3 is 3. In some embodiments, n3 is 5. In some embodiments, n3 is 7. In some embodiments, n4 is 1. In some embodiments, n4 is 3. In some embodiments, n4 is 5. In some embodiments, n4 is 7. In some embodiments, n5 is 1. In some embodiments, n5 is 3. In some embodiments, n5 is 5. In some embodiments, n5 is 7. In certain embodiments, two of n3, n4, and n5 are 1. In certain embodiments, all three of n3, n4, and n5 are 1.

[0084] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, And this compound has the formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0085] In another context, compounds of formula (XXX), as well as their salts, solvates, and hydrates, are provided: [ka] In equation (XXX): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is an oligonucleotide sequence; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0086] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0087] In another context, oligonucleotides comprising the part of formula (XVII), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XVII): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R is an oligonucleotide; R' is either O or S; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O) , CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0088] In another context, oligonucleotides comprising the part of formula (XVII-a), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XVII-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is a binding to the oligonucleotide; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0089] In another context, oligonucleotides comprising the part of formula (XVIII), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XVIII): R 9is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is a combination; Y1 is O; Y2 is O; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; n2 is 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0090] In another context, oligonucleotides comprising the part of formula (XVIII-a), as well as its salts, solvates, and hydrates, are provided: [ka] In equation (XVIII-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; L is a bond, phosphodiester bond, phosphorothioate bond, triazole, tetrazole, amide, reverse amide, carbamate, carbonate, urea, alkyl, or heteroalkyl; R 2 This is a binding to the oligonucleotide; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2.

[0091] In some embodiments, R 9 is H, adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each containing a protecting group (PG). In some embodiments, R 9 R is a modified nucleic acid base. In some embodiments, R 9 These are alkyl groups, aryl groups, or heteroaryl groups, which are substituted as needed.

[0092] In another context, compounds of formula (XIX), or salts, solvates, or hydrates thereof are provided: [ka] In equation (XIX): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is H, a protecting group (PG), or [ka] and; X 1 teeth, [ka] [ka] and X2 is H, alkyl, or a protecting group (PG).

[0093] In another context, compounds of formula (XX), or their salts, solvates, or hydrates are provided: [ka] In equation (XX): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is H, a protecting group (PG), or [ka] and; X 2 is H, halogen, OH, O-alkyl, O-heteroalkyl, or O-protecting group (PG); X 3 is O, CH2, CH2O, or NH as needed; X 4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; and Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0094] In another context, compounds of formula (XXI), or their salts, solvates, or hydrates are provided: [ka] In equation (XXI): R 9is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is H, a protecting group (PG), or [ka] and; X 2 is H, halogen, OH, O-alkyl, O-heteroalkyl, or protecting group (PG); X 3 is O, CH2, CH2O, or NH as needed; X 4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; and Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0095] In another context, compounds of formula (XXII), or their salts, solvates, or hydrates are provided: [ka] In equation (XXII): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is H, a protecting group (PG), or [ka] and; X 2is H, halogen, OH, O-alkyl, O-heteroalkyl, or O-protecting group (PG); X 3 is O, CH2, CH2O, or NH as needed; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0096] In another context, compounds of formula (XXIII), or their salts, solvates, or hydrates are provided: [ka] In equation (XXIII): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 2 is H, a protecting group (PG), or [ka] and; X 2 is H, halogen, OH, O-alkyl, O-heteroalkyl, or O-protecting group (PG); X 3 is O, CH2, CH2O, or NH as needed; X 4 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; and Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0097] In some embodiments, this compound: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or its salt, solvate, or hydrate.

[0098] In another context, compounds of formula (XXIV), or their salts, solvates, or hydrates are provided: [ka] In equation (XXIV): R9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; X 2 is H, alkyl, or a protecting group (PG); X 5 is H, a protecting group (PG), or [ka] and X 6 teeth, [ka] That is the case.

[0099] In another context, compounds of formula (XXV), or their salts, solvates, or hydrates are provided: [ka] In equation (XXV): R 9 is adenine, guanine, thymine, cytosine, or uracil, or adenine, guanine, thymine, cytosine, or uracil, each comprising a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl; X 2 is H, OH, O-alkyl, alkyl, heteroalkyl, halogen, or O-protecting group (PG); X 5 is an H, OH, O-protecting group (PG), or [ka] and; X 7 is O; X8 is CO, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; and Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, or 6.

[0100] In some embodiments, this compound: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, this compound is: [ka] or a salt, solvate, or hydrate thereof. In some embodiments, any of the compounds provided herein may be bound to an oligonucleotide. In some embodiments, any of the compounds provided herein may be deprotected (e.g., the acetyl group on the oxygen atom in GalNAc may be removed) and then bound to an oligonucleotide. In some embodiments, any of the compounds provided herein, or an oligonucleotide containing any of the compounds provided herein, may be provided in a composition containing a pharmaceutically acceptable carrier. In some embodiments, any of the compounds, oligonucleotides, or compositions provided herein may be administered to a subject in a manner for modulating protein function in the subject. In some embodiments, any of the compounds, oligonucleotides, or compositions provided herein may be administered to a subject in a manner for treating or improving a disease, disorder, or symptom thereof in the subject. In some embodiments, the disease, disorder, or symptom thereof is a disease, disorder, or symptom thereof of the liver.

[0101] In another aspect, the Disclosure provides methods for producing any of the compounds provided herein, including one or more compounds and chemical transformations described herein, including the examples herein, for example, Examples 1 to 23.

[0102] In another context, the present disclosure relates to a compound of formula (XXVI), or a salt, solvate, or hydrate thereof: [ka] In equation (XXVI): Each n is independently 1, 2, 3, 4, or 5; Each m is independently 0, 1, 2, 3, 4, 5, or 6; Each o is independently 0, 1, 2, 3, 4, 5, or 6; Each Y1 independently has O, CH(R a ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Y2 independently has O, CH(R b ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Het1, Het2, and Het3 are each independently a heteroaryl or heterocycline as needed; Each R1 can independently be alkyl-O-phosphoramidite, optionally substituted alkenylphosphoramidite, optionally substituted alkynylphosphoramidite, OH, NH2, or NHR a N3, C(=O)OH, C(=O)X, CN, SH, SSH, SO2X, C(=O)NHNH2, NHNH2, C(=S)NHNH2, C(=S)NH2, NHOH, C(=O)CH2X, malonyl, alkyl, alkenyl, dienyl, alkynyl, heteroalkyl, -OP(=S)X, -C(=O)H, -C(=O)R a -N=C=O, -N=C=NR a -N=C=S, CHX, -OP(=O)OH, phosphane, alkoxyphosphane, -C(R a )2. A therapeutic agent for regulating Michael acceptor, protein, or hepatocytes, or for treating liver disease, where X is a leaving group; R5, R6, and R7 are each independent of each other. [ka] and; R9 is a heterocyclyl that is substituted as needed; Each R a These are independently H, alkyl, halo, OR c , or SR c and; Each R b These are independently H, alkyl, halo, OR c , or SR c and Each R c A compound, its salt, solvate, or hydrate, in which is independently H or alkyl; Furthermore, compositions containing such compounds, as well as their salts, solvates, and hydrates. The present invention provides methods for treatment including these, and methods for production including these.

[0103] In one aspect of the present invention, each R1 is independently an alkyl-O-phosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite.

[0104] In one aspect of the present invention, each R1 is independently OH, NH2, NHR a N3, C(=O)OH, C(=O)X, CN, SH, SSH, SO2X, C(=O)NHNH2, NHNH2, C(=S)NHNH2, C(=S)NH2, NHOH, C(=O)CH2X, malonyl, alkyl, alkenyl, dienyl, alkynyl, heteroalkyl, -OP(=S)X, -C(=O)H, -C(=O)R a -N=C=O, -N=C=NR a -N=C=S, CHX, -OP(=O)OH, phosphane, alkoxyphosphane, or -C(R a )2, where X is a leaving group.

[0105] In one aspect of the present invention, each R1 is independently a therapeutic agent for modulating Michael acceptors, proteins, or hepatocytes, or for treating liver diseases.

[0106] In another context, a compound of formula (XXVI-a), or a salt, solvate, or hydrate thereof: [ka] In equation (XXVI-a): Each n is independently 1, 2, 3, 4, or 5; Each m is independently 0, 1, 2, 3, 4, 5, or 6; Each o is independently 0, 1, 2, 3, 4, 5, or 6; Each of L1, L2, and L3 is independent of either not existing, C(=O), or C(=O)NH; Each Y1 independently has O, CH(R a), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Y2 independently has O, CH(R b ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Het1, Het2, and Het3 are each independently a heteroaryl or heterocycline as needed; Each R1 independently consists of an alkyl-O-phosphoramidite, optionally substituted alkenylphosphoramidite, optionally substituted alkynylphosphoramidite, and O H, NH2, NHR a N3, CH2N3, C(=O)OH, C(=O)X, CN, SH, SSH, SO2X, C(=O)NHNH2, NHNH2, C(=S)NHNH2, C(=S)NH2, NHOH, C(=O)CH2X, malonyl, alkyl, alkenyl, dienyl, alkynyl, heteroalkyl, -OP(=S)X, -C(=O)H, -C(=O)R a -N=C=O, -N=C=NR a -N=C=S, CHX, -OP(=O)OH, phosphate, phosphane, alkoxyphosphane, -C(R a )2, a therapeutic agent for modulating Michael acceptor, protein, or hepatocytes, or for treating liver disease, where X is a leaving group; or R 1This includes oligonucleotides linked by bonds, phosphodiester bonds, phosphorothioate bonds, triazoles, tetrazoles, amides, reverse amides, carbamates, carbonates, ureas, alkyls, or heteroalkyls; R5, R6, and R7 are each independent of each other. [ka] and; R9 is a heterocyclyl that is substituted as needed; Each R a These are independently H, alkyl, halo, OR c , or SR c and; Each R b These are independently H, alkyl, halo, OR c , or SR c and Each R c A compound, its salt, solvate, or hydrate, in which is independently H or alkyl; The present invention also provides compositions containing such compounds, as well as salts, solvates, and hydrates thereof, methods of treatment containing these, and methods of preparation containing these.

[0107] In some embodiments, R9 is a nitrogen-containing heterocycline, optionally substituted. In some embodiments, R9 is a pyrimidinyl, optionally substituted. In some embodiments, R9 is 4H-1λ 2 ,3λ 2 -Pyrimidine-2,4-dione. In certain embodiments, R9 is [ka] That is the case.

[0108] In some embodiments, each of Het1, Het2, and Het3 is independently a nitrogen-containing heterocyclyl or nitrogen-containing heteroaryl, as optionally substituted. In some embodiments, each of Het1, Het2, and Het3 is independently a 1,2,3-triazolyl, as optionally substituted. In some embodiments, each of Het1, Het2, and Het3 is independently a 1λ, as optionally substituted. 2 ,2,3-triazole-4-yl. Specific embodiment In this case, each of Het1, Het2, and Het3 operates independently. [ka] That is the case.

[0109] In some embodiments, two of R5, R6, and R7 are the same. In some embodiments, R5 and R6 are the same. In some embodiments, three of R5, R6, and R7 are the same. In some embodiments, o is the same in each of R5, R6, and R7. In a particular embodiment, o is the same in each of R5 and R6. In a particular embodiment, all three of R5, R6, and R7 are [ka] In a particular embodiment, all three R5, R6, and R7 are, [ka] That is the case.

[0110] In some embodiments of the present invention, each R1 is independently an alkyl-O-phosphoramidite, optionally substituted alkenylphosphoramidite, or optionally substituted alkynylphosphoramidite. In some embodiments of the present invention, each R1 is independently OH, NH2, NHR aN3, C(=O)OH, C(=O)X, CN, SH, SSH, SO2X, C(=O)NHNH2, NHNH2, C(=S)NHNH2, C(=S)NH2, NHOH, C(=O)CH2X, malonyl, alkyl, alkenyl, dienyl, alkynyl, heteroalkyl, -OP(=S)X, -C(=O)H, -C(=O)R a -N=C=O, -N=C=NR a -N=C=S, CHX, -OP(=O)OH, phosphane, alkoxyphosphane, or -C(R a )2, where X is a leaving group. In some embodiments of the present invention, each R1 is independently a therapeutic agent for modulating Michael acceptors, proteins, or hepatocytes, or for treating liver diseases. In certain embodiments, R1 is [ka] In certain embodiments, R1 is a phosphate. In certain embodiments, R1 is CH2N3.

[0111] In another context, the compound of formula (XXVII), or its salt, solvate, or water Japanese food is offered: [ka] In equation (XXVII), Each n is independently 1, 2, 3, 4, or 5; Each m is independently 0, 1, 2, 3, 4, 5, or 6; Each o is independently 0, 1, 2, 3, 4, 5, or 6; Each Y1 independently has O, CH(R a ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Y2 independently has O, CH(R b ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Het1, Het2, and Het3 are each independently a heteroaryl or heterocycline as needed; R5, R6, and R7 are each independent of each other. [ka] and; R9 is a heterocyclyl that is substituted as needed; and R 2 H, protecting group (PG), [ka] and; Each R a These are independently H, alkyl, halo, OR c , or SR c and; Each R b These are independently H, alkyl, halo, OR c , or SR c and Each R c These are independently either H or alkyl.

[0112] In some embodiments, R 2 is H. In some embodiments, R 2 R is a protecting group (PG). In certain embodiments, R 2 teeth, [ka] That is the case.

[0113] In some embodiments, R9 is a nitrogen-containing heterocycline, optionally substituted. In some embodiments, R9 is a dihydropyrimidinyl, optionally substituted tetrahydropyrimidinyl, or optionally substituted hexahydropyrimidinyl. In some embodiments, R9 is 4H-1λ 2 ,3λ 2 -Pyrimidine-2,4-dione. In certain embodiments, R9 is [ka] That is the case.

[0114] In some embodiments, each of Het1, Het2, and Het3 is independently a nitrogen-containing heterocyclyl or nitrogen-containing heteroaryl, as optionally substituted. In some embodiments, each of Het1, Het2, and Het3 is independently a 1,2,3-triazolyl, as optionally substituted. In some embodiments, each of Het1, Het2, and Het3 is independently a 1λ, as optionally substituted. 2 It is 2,3-triazole-4-yl. In certain embodiments, each of Het1, Het2, and Het3 is independently [ka] That is the case.

[0115] In some embodiments, two of R5, R6, and R7 are the same. In some embodiments, R5 and R6 are the same. In some embodiments, three of R5, R6, and R7 are the same. In some embodiments, o is the same in each of R5, R6, and R7. In a particular embodiment, o is the same in each of R5 and R6.

[0116] In some embodiments, this compound is of formula (XXVII-a): [ka] It is a compound of [the compound].

[0117] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0118] In another context, this disclosure relates to a portion of formula (XXVIII), or its salts, solvates, or hydrates: [ka] We provide a modified oligonucleotide containing, in formula (XXVIII): Each n is independently 0, 1, 2, 3, 4, or 5; Each m is independently 0, 1, 2, 3, 4, 5, or 6; Each o is independently 0, 1, 2, 3, 4, 5, or 6; Each Y1 independently has O, CH(R a ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Y2 independently has O, CH(R b ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Het1, Het2, and Het3 are each independently a heteroaryl or heterocycline as needed; R5, R6, and R7 are each independent of each other. [ka] and R9 is a heterocyclyl that is substituted as needed; L is a bond, O, CH2, S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, alkyl, alkenyl, dienyl, alkynyl, heteroalkyl, phosphate, or thiol-Michael adduct; Each R a These are independently H, alkyl, halo, OR c, or SR c and; Each R b These are independently H, alkyl, halo, OR c , or SR c and Each R c These are independently either H or alkyl.

[0119] In another context, this disclosure relates to a portion of formula (XXIX), or its salts, solvates, or hydrates: [ka] We provide an oligonucleotide containing, in formula (XXIX): Each n is independently 0, 1, 2, 3, 4, or 5; Each m is independently 0, 1, 2, 3, 4, 5, or 6; Each o is independently 0, 1, 2, 3, 4, 5, or 6; Each Y1 independently has O, CH(R a ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Each Y2 independently has O, CH(R b ), S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)(OH)-O-, P(=O)(SH)-O, P(=S)(SH)-O, -OP(=O)(OH)-O-, -OP(=O)(SH)-O-, -OP(=S)(SH)-O-, -OP(=O)(OH)-, -OP(=O)(SH)-, -OP(=S)(SH)-; Het1, Het2, and Het3 are each independently a heteroaryl or heterocycline as needed; R5, R6, and R7 are each independent of each other. [ka] and; R9 is a heterocyclyl that is substituted as needed; L is a bond, O, CH2, S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, alkyl, alkenyl, dienyl, alkynyl, heteroalkyl, phosphate, or thiol-Michael adduct; Each R a These are independently H, alkyl, halo, OR c , or SR c and; Each R b These are independently H, alkyl, halo, OR c , or SR c and Each R c These are independently either H or alkyl.

[0120] In some embodiments, this oligonucleotide includes siRNA, miRNA, ADAR mobilization molecules, ADAR targeting molecules, guide RNA, or antisense nucleic acids. In some embodiments, any of the compounds or oligonucleotides provided herein, or their salts, solvates, or hydrates, may be used as reagents in chemical reactions.

[0121] Compositions containing any of the compounds provided herein and a pharmaceutically acceptable carrier are also provided herein.

[0122] Methods for producing any of the compounds disclosed herein are further provided herein, and these methods include one or more of the compounds and chemical transformations described herein, including the examples herein, for example, Examples 1 to 23.

[0123] In some embodiments, this compound is a compound of formula (XXXI), or a salt, solvate, or hydrate thereof: [ka] And in equation (XXXI): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 H, O-protecting group (PG), [ka] , linker, azide, carboxylic acid, or amine; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, C(O)NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y5 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y6 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0124] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0125] In some embodiments, this compound is a compound of formula (XXXII), or a salt, solvate, or hydrate thereof: [ka] And in equation (XXXII): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 is an H, O-protecting group (PG), or [ka] , linker, azide, carboxylic acid, or amine; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y5 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y6 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y7 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0126] In some embodiments, this compound is given by formula: [ka] It is a compound of [the compound].

[0127] In some embodiments, this compound is a compound of formula (XXXII-a), or a salt, solvate, or hydrate thereof: [ka] And in equation (XXXII-a): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 is an H, O-protecting group (PG), or [ka] , linker, azide, carboxylic acid, or amine; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y5 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y6 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y7 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, and n5 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0128] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof. In some embodiments, this compound is of formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0129] In some embodiments, this compound is a compound of formula (XXXIII), or a salt, solvate, or hydrate thereof: [ka] And in equation (XXXIII): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 is an H, O-protecting group (PG), or [ka] , linker, azide, carboxylic acid, or amine; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y4 is CO, C(O)NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y5 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y6 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; n2 is 0, 1, 2, 3, 4, 5, or 6; Each of n1, n3, n4, n5, n6, n7, and n8 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0130] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0131] In some embodiments, this compound is a compound of formula (XXXIV), or a salt, solvate, or hydrate thereof: [ka] And in equation (XXXIV): R 9 H, adenine, guanine, thymine, cytosine, or uracil, or each containing a protecting group (PG), a modified nucleic acid base, optionally substituted alkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted carbocykrill, optionally substituted heterocyclyl, or nucleic acid base isocouples; R 2 H, O-protecting group (PG), [ka]

[0132] , linker, azide, carboxylic acid, or amine; Y1 is O, CH2, CH2O, or NH as needed; Y2 is O, CH2, CH2O, or NH as needed; Y3 is CO, C(O)NH, C(O)-NH-CH2-C(O)-NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or It is CH2; Y4 is CO, C(O)NH, SO2, P(O)O, CH2-OC(O), CH2-NH-C(O), CH2-NH-SO2, or CH2; Y5 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Y6 is CO, C(O)NH, OC(O)NH, or OCH2C(O)NH; Each of n1, n2, and n3 is independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0133] In some embodiments, this compound is given by formula: [ka] It is a compound of, or a salt, solvate, or hydrate thereof.

[0134] In one aspect, the compounds herein have one or more GalNAc moieties, and these GalNAc moieties: [ka] It is a compound of any of the formulas illustrated herein.

[0135] In one aspect, the compounds herein have one or more acetylated GalNAc moieties, and these acetylated GalNAc moieties: [ka] It is a compound of any of the formulas illustrated herein.

[0136] In one aspect, the compounds herein have one or more protected (PG)GalNAc moieties, and these protected GalNAc moieties: [ka] It is a compound of any of the formulas illustrated herein.

[0137] In one aspect, the oligonucleotide-containing compounds described herein have one or more GalNAc moieties, and these GalNAc moieties: [ka] It is a compound of any of the formulas illustrated herein.

[0138] In one aspect, the oligonucleotide-containing compounds in this specification have one or more acetylated GalNAc moieties, and this acetylated GalNAc moiety: [ka] It is a compound of any of the formulas illustrated herein.

[0139] In one aspect, the oligonucleotide-containing compounds described herein have one or more protected (PG)GalNAc moieties, and these protected GalNAc moieties: [ka] It is a compound of any of the formulas illustrated herein.

[0140] Compounds having all free hydroxyl groups on the GalNAc moiety can be obtained by using chemical synthesis techniques known in the art (e.g., deprotection; removal of protecting groups (PG)) to selectively deacetylate the acetyl moiety on the oxygen atom of the GalNAc structure while simultaneously leaving the acetyl group bonded to the nitrogen atom of the GalNAc structure intact. [Brief explanation of the drawing]

[0141] [Figure 1] Figure 1 shows the 1H NMR spectrum of product 2 from Example 3.

[0142] [Figure 2] Figure 2 shows the 1H NMR spectrum of product 3 from Example 3.

[0143] [Figure 3]Figure 3 shows the 1H NMR spectrum of product 4 from Example 3.

[0144] [Figure 4] Figure 4 shows the 1H NMR spectrum of product 6 from Example 3.

[0145] [Figure 5] Figure 5 shows the 1H NMR spectrum of product 7 from Example 3.

[0146] [Figure 6] Figure 6 shows the 1H NMR spectrum of product 8 from Example 3.

[0147] [Figure 7] Figure 7 shows the 1H NMR spectrum of product 10 from Example 3.

[0148] [Figure 8] Figure 8 shows the tandem mass spectrometry of product 10 from Example 3.

[0149] [Figure 9] Figure 9 shows the tandem mass spectrometry of product 11 from Example 3.

[0150] [Figure 10] Figure 10 shows the 1H NMR spectrum of product 11 from Example 3.

[0151] [Figure 11] Figure 11 shows the 13C NMR spectrum of product 11 from Example 3.

[0152] [Figure 12] Figure 12 shows the 1H NMR spectrum of product 2 from Example 4.

[0153] [Figure 13] Figure 13 shows the 1H NMR spectrum of product 3 from Example 4.

[0154] [Figure 14] Figure 14 shows the 1H NMR spectrum of product 4 from Example 4.

[0155] [Figure 15] Figure 15 shows the 1H NMR spectrum of product 5 from Example 4.

[0156] [Figure 16] Figure 16 shows the 1H NMR spectrum of product 6 from Example 4.

[0157] [Figure 17] Figure 17 shows the 1H NMR spectrum of product 7 from Example 4.

[0158] [Figure 18] Figure 18 shows the 1H NMR spectrum of product 8 from Example 4.

[0159] [Figure 19] Figure 19 shows the 1H NMR spectrum of product 10 from Example 4.

[0160] [Figure 20] Figure 20 shows the tandem mass spectrometry of product 10 from Example 4.

[0161] [Figure 21] Figure 21 shows the 1H NMR spectrum of compound 6 from Example 7.

[0162] [Figure 22] Figure 22 shows the 31P NMR spectrum of compound 5 from Example 7.

[0163] [Figure 23] Figure 23 shows the 1H NMR spectrum of compound 2 from Example 7.

[0164] [Figure 24] Figure 24 shows the 1H NMR spectrum of compound 3 from Example 7.

[0165] [Figure 25] Figure 25 shows the 1H NMR spectrum of GalNAc-[PEG]4-NH2 trifluoroacetate from Example 7.

[0166] [Figure 26] Figure 26 shows the 1H NMR spectrum of compound 10 from Example 7.

[0167] [Figure 27] Figure 27 shows the 1H NMR spectrum of compound 4 from Example 7.

[0168] [Figure 28] Figure 28 shows the 1H NMR spectrum of compound 2 from Example 8.

[0169] [Figure 29] Figure 29 shows the 1H NMR spectrum of compound 3 from Example 8.

[0170] [Figure 30] Figure 30 shows the 1H NMR spectrum of compound 4 from Example 8.

[0171] [Figure 31] Figure 31 shows the 1H NMR spectrum of compound 5 from Example 8.

[0172] [Figure 32] Figure 32 shows the 1H NMR spectrum of compound 6 from Example 8.

[0173] [Figure 33] Figure 33 shows the 1H NMR spectrum of compound 8 from Example 8.

[0174] [Figure 34] Figure 34 shows the 1H NMR spectrum of compound 9 from Example 8.

[0175] [Figure 35] Figure 35 shows the 1H NMR spectrum of compound 12 from Example 8.

[0176] [Figure 36] Figure 36 shows the 1H NMR spectrum of compound 10 from Example 8.

[0177] [Figure 37] Figure 37 shows the 1H NMR spectrum of compound HA-103 from Example 8.

[0178] [Figure 38] Figure 38 shows the 31P NMR spectrum of compound HA-103 from Example 8.

[0179] [Figure 39] Figure 39 shows the 1H NMR spectrum of compound 2 from Example 9.

[0180] [Figure 40] Figure 40 shows the 1H NMR spectrum of compound 3 from Example 9.

[0181] [Figure 41] Figure 41 shows the 1H NMR spectrum of compound 4 from Example 9.

[0182] [Figure 42] Figure 42 shows the 1H NMR spectrum of compound 5 from Example 9.

[0183] [Figure 43] Figure 43 shows the 1H NMR spectrum of compound 6 from Example 9.

[0184] [Figure 44] Figure 44 shows the 1H NMR spectrum of compound 7 from Example 9.

[0185] [Figure 45] Figure 45 shows the 1H NMR spectrum of compound 9 from Example 9.

[0186] [Figure 46] Figure 46 shows the 1H NMR spectrum of compound 10 from Example 9.

[0187] [Figure 47] Figure 47 shows the 31P NMR spectrum of compound 10 from Example 9.

[0188] [Figure 48] Figure 48 shows the 1H NMR spectrum of compound 11 from Example 10.

[0189] [Figure 49] Figure 49 shows the 1H NMR spectrum of compound 10 from Example 10.

[0190] [Figure 50] Figure 50 shows the 1H NMR spectrum of compound H4-(HA-111) from Example 10.

[0191] [Figure 51] Figure 51 shows the 31P NMR spectrum of compound H4-(HA-111) from Example 10.

[0192] [Figure 52] Figure 52 shows the 1H NMR spectrum of compound 12 from Example 11.

[0193] [Figure 53] Figure 53 shows the 1H NMR spectrum of compound H6-(HA-112) from Example 11.

[0194] [Figure 54] Figure 54 shows the 31P NMR spectrum of compound H6-(HA-112) from Example 11.

[0195] [Figure 55] Figure 55 shows the 1H NMR spectrum of compound 2 from Example 12.

[0196] [Figure 56] Figure 56 shows the 1H NMR spectrum of compound 3 from Example 12.

[0197] [Figure 57] Figure 57 shows the 1H NMR spectrum of compound 4 from Example 12.

[0198] [Figure 58] Figure 58 shows the 1H NMR spectrum of compound 5 from Example 12.

[0199] [Figure 59] Figure 59 shows the 1H NMR spectrum of compound 6 from Example 12.

[0200] [Figure 60] Figure 60 shows the 1H NMR spectrum of compound H6-(HA-113) from Example 12.

[0201] [Figure 61] Figure 61 shows the 31P NMR spectrum of compound H6-(HA-113) from Example 12.

[0202] [Figure 62] Figure 62 shows the 1H NMR spectrum of compound 2 from Example 13.

[0203] [Figure 63] Figure 63 shows the 1H NMR spectrum of compound 3 from Example 13.

[0204] [Figure 64] Figure 64 shows the 1H NMR spectrum of compound 4 from Example 13.

[0205] [Figure 65] Figure 65 shows the 1H NMR spectrum of compound 5 from Example 13.

[0206] [Figure 66] Figure 66 shows the 1H NMR spectrum of compound 7 from Example 13.

[0207] [Figure 67] Figure 67 shows the 1H NMR spectrum of compound HA-114 from Example 13.

[0208] [Figure 68] Figure 68 shows the 31P NMR spectrum of compound HA-114 from Example 13.

[0209] [Figure 69] Figure 69 shows the 1H NMR spectrum of compound 3 from Example 14.

[0210] [Figure 70] Figure 70 shows the 1H NMR spectrum of compound 5 from Example 14.

[0211] [Figure 71] Figure 71 shows the 1H NMR spectrum of compound H8-(HA-115) from Example 14.

[0212] [Figure 72] Figure 72 shows the 31P NMR spectrum of compound H8-(HA-115) from Example 14.

[0213] [Figure 73] Figure 73 shows the 1H NMR spectrum of compound 9 from Example 15.

[0214] [Figure 74] Figure 74 shows the 1H NMR spectrum of compound 10 from Example 15.

[0215] [Figure 75] Figure 75 shows the 1H NMR spectrum of compound 12 from Example 15.

[0216] [Figure 76] Figure 76 shows the 1H NMR spectrum of compound 13 from Example 15.

[0217] [Figure 77] Figure 77 shows the 1H NMR spectrum of compound 14 from Example 15.

[0218] [Figure 78] Figure 78 shows the 1H NMR spectrum of compound H9-(HA-116) from Example 15.

[0219] [Figure 79] Figure 79 shows the 31P NMR spectrum of compound H9-(HA-116) from Example 15.

[0220] [Figure 80] Figure 80 shows the 1H NMR spectrum of compound 2 from Example 16.

[0221] [Figure 81] Figure 81 shows the 1H NMR spectrum of compound 6 from Example 16.

[0222] [Figure 82] Figure 82 shows the 1H NMR spectrum of compound 8 from Example 16.

[0223] [Figure 83]Figure 83 shows the 1H NMR spectrum of compound 12 from Example 16.

[0224] [Figure 84] Figure 84 shows the 1H NMR spectrum of compound 14 from Example 16.

[0225] [Figure 85] Figure 85 shows the 1H NMR spectrum of compound 15(H9-(HA-118)) from Example 16.

[0226] [Figure 86] Figure 86 shows the 3P NMR spectrum of compound 15 (H9-(HA-118)) from Example 16.

[0227] [Figure 87] Figure 87 shows the 1H NMR spectrum of compound 2 from Example 17.

[0228] [Figure 88] Figure 88 shows the 1H NMR spectrum of compound 5 from Example 17.

[0229] [Figure 89] Figure 89 shows the 1H NMR spectrum of compound 8 from Example 17.

[0230] [Figure 90] Figure 90 shows the 1H NMR spectrum of compound 9 from Example 17.

[0231] [Figure 91] Figure 91 shows the 1H NMR spectrum of compound 13 from Example 17.

[0232] [Figure 92] Figure 92 shows the 1H NMR spectrum of compound 14 from Example 17.

[0233] [Figure 93] Figure 93 shows the 1H NMR spectrum of compound 15 (Hd-(HA-121)) from Example 17.

[0234] [Figure 94] Figure 94 shows the 3P NMR spectrum of compound 15 (Hd-(HA-121)) from Example 17. [Modes for carrying out the invention]

[0235] Detailed explanation definition For the purpose of making the present invention easier to understand, certain terms are defined here first for convenience.

[0236] As used herein, the term “to treat” a disorder includes improving, mitigating, and / or managing the disorder and / or any condition that may cause the disorder. The terms “to treat” and “treatment” mean any method of alleviating or mitigating a disease and / or its associated symptoms. As used herein, “to treat” includes blocking, inhibiting, attenuating, protecting from, modulating, reversing its effects, and, for example, reducing the occurrence of the adverse effects of the disorder. As used herein, “inhibit” includes preventing, reducing, and halting the progression of the disorder.

[0237] The terms “isolated,” “purified,” or “biologically pure” refer to a material that is substantially or essentially free from components that would normally be associated with it if found in its state of origin. Purity and homogeneity are typically determined using analytical scientific techniques such as polyacrylamide gel electrophoresis or high-performance liquid chromatography (HPLC). In particular, in certain embodiments, the compound or oligonucleotide is at least 85% pure, more preferably at least 90% pure, more preferably at least 95% pure, and most preferably at least 99% pure.

[0238] The term “administer” or “to administer” encompasses the route by which a compound or oligonucleotide is introduced into a subject in order to perform its intended function. Examples of possible routes of administration include injection (subcutaneous, intravenous, parenteral, intraperitoneal, intrasacral), topical, oral, inhalation, rectal, and percutaneous.

[0239] The term "effective dose" encompasses the amount that is effective in achieving the desired outcome at the required dosage and duration. The effective dose of a compound or oligonucleotide may vary depending on factors such as the subject's disease state, age, and weight, as well as the compound or oligonucleotide's ability to elicit the desired response in the subject. The dosage regimen may be adjusted to provide the optimal therapeutic response. The effective dose is also the amount at which the therapeutically beneficial effects outweigh any toxic or adverse effects (e.g., side effects) of the inhibitor compound or oligonucleotide.

[0240] The terms “systemic administration,” “administered systemically,” “administered peripherally,” and “administered peripherally” mean, as used herein, that a compound, oligonucleotide, drug, or other substance enters the patient’s entire body and is therefore subjected to processes such as metabolism. This means administering these substances.

[0241] The term "therapeutic dose" refers to the amount of compound or oligonucleotide administered that is sufficient to prevent, to some extent, the onset of one or more symptoms of the condition or disorder being treated, or to alleviate them.

[0242] The therapeutically effective dose (i.e., effective dosage) of the compound or oligonucleotide may range from about 0.005 μg / kg to about 200 mg / kg, preferably from about 0.01 mg / kg to about 200 mg / kg, and more preferably from about 0.015 mg / kg to about 30 mg / kg body weight. In other embodiments, the therapeutically effective dose may range from about 1.0 pM to about 10 μM. Those skilled in the art will understand that certain factors may influence the dosage required to effectively treat a subject, including, but not limited to, the severity of the disease or disorder, previous treatments, the subject's general health and / or age, and other pre-existing conditions. Furthermore, treatment of a subject with a therapeutically effective dose of the compound or oligonucleotide may consist of a single treatment or, preferably, a series of treatments. In one example, a subject is treated once daily with a compound or oligonucleotide in the range of approximately 0.005 μg / kg to approximately 200 mg / kg body weight for approximately 1 to 10 weeks, preferably 2 to 8 weeks, more preferably about 3 to 7 weeks, and even more preferably about 4, 5, or 6 weeks. In another example, a subject may be treated daily for several years in the context of a chronic condition or disease. It is also understood that the effective dosage of the compound or oligonucleotide used for treatment may be increased or decreased over the course of a particular treatment.

[0243] The term "chiral" refers to a molecule that has the property of not being able to be superimposed on its mirror image partner, while the term "achiral" refers to a molecule that can be superimposed on its mirror image partner.

[0244] The term "diastereomer" refers to stereoisomers that have two or more asymmetric centers, and in which these molecules are not mirror images of each other.

[0245] The term "enantiomer" refers to two stereoisomers of a compound that are mirror images of each other but cannot be superimposed. An equimolar mixture of two enantiomers is called a "racemic mixture" or "racemate."

[0246] The terms "isomer" or "stereoisomer" refer to compounds that have the same chemical structure but differ in the arrangement of atoms or groups in space.

[0247] The term "prodrug" encompasses compounds that have a moiety that can be metabolized in vivo. Generally, prodrugs are metabolized in vivo to an active drug by an esterase or other mechanism. Examples of prodrugs and their uses are well known in the art (see, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19). Prodrugs can be prepared in situ during the final isolation and purification of the compound, or by reacting a purified compound in free acid form or as a hydroxyl group separately with a suitable esterifying agent. The hydroxyl group can be converted to an ester by treatment with a carboxylic acid. Examples of prodrug moieties include substituted and unsubstituted, branched or unbranched, lower alkyl ester moieties (e.g., propionic acid esters), lower alkenyl esters, di-lower alkyl-amino lower alkyl esters (e.g., dimethylaminoethyl esters), acrylamino lower alkyl esters (e.g., acetyloxymethyl esters), and acyloxy lower alkyl esters (e.g., pivaloyloxymethyl esters). Examples include aryl esters (phenyl esters), aryl-lower alkyl esters (e.g., benzyl esters), substituted (e.g., with methyl, halo, or methoxy substituents) aryl and aryl-lower alkyl esters, amides, lower alkylamides, di-lower alkylamides, and hydroxyamides. Preferred prodrug moieties are propionic acid esters and acyl esters. Prodrugs that are converted to an active form in vivo by other mechanisms are also included. In some aspects, the compounds of this disclosure are prodrugs of any of the formulas herein.

[0248] The term "subject" refers to animals such as mammals (including, but not limited to, primates (e.g., humans), cattle, sheep, goats, horses, dogs, cats, rabbits, rats, and mice). In certain embodiments, the subject is a human.

[0249] The terms “a,” “an,” and “the,” when used in this application, including in the claims, mean “one or more.” Therefore, for example, a reference to “a sample” includes multiple samples unless the context clearly indicates otherwise (e.g., multiple samples).

[0250] Throughout this specification and the claims, the words “comprise,” “comprises,” and “comprising” are used in a non-exclusive sense unless the context requires otherwise.

[0251] As used herein, the term “about” means, when referring to a value, to include variations from the specified amount, such as ±20% in some embodiments, ±10% in some embodiments, ±5% in some embodiments, ±1% in some embodiments, ±0.5% in some embodiments, and ±0.1% in some embodiments. Such variations are appropriate for carrying out the disclosed method or employing the disclosed composition.

[0252] As used herein, the term “alkyl” means a linear or branched hydrocarbon group containing 1 to 20 carbon atoms. Examples of specified numbers of carbon atoms within this range include C1-C12 alkyls (containing 1 to 12 carbon atoms) and C1-C4 alkyls (containing 1 to 4 carbon atoms). The term “lower alkyl” means a C1-C6 alkyl chain. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, tert-butyl, and n-pentyl. Alkyl groups may be substituted with one or more substituents as needed.

[0253] The term "haloalkyl" refers to an alkyl group substituted with one or more halo substituents. Examples of haloalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, bromomethyl, chloromethyl, and 2,2,2-trifluoroethyl.

[0254] The term "alkenyl" refers to an unsaturated hydrocarbon chain, which may be linear or branched, containing 2 to 12 carbon atoms and at least one carbon-carbon double bond. The alkenyl group may be substituted with one or more substituents as needed.

[0255] The term "aryl alkenyl" refers to a linear or branched chain containing 2 to 12 carbon atoms and at least one carbon-carbon double bond, and whose alkenyl unit is sp 2 This refers to an unsaturated hydrocarbon chain in which one or more of the hybridized carbon atoms are bonded to the aryl moiety. The alkenyl group may be substituted with one or more substituents as needed.

[0256] The term "alkynyl" refers to an unsaturated hydrocarbon chain, which may be linear or branched, containing 2 to 12 carbon atoms and at least one carbon-carbon triple bond. The alkynyl group may be substituted with one or more substituents as needed.

[0257] The term "arylalkynyl" refers to an unsaturated hydrocarbon chain, which may be linear or branched, containing 2 to 12 carbon atoms and at least one carbon-carbon triple bond, with one or more of the sp hybrid carbons of the alkynyl unit bonded to the aryl portion. The alkynyl group may be substituted with one or more substituents as needed.

[0258] sp 2 Alternatively, the sp carbon may, if necessary, be a bonding site for its alkenyl and alkynyl groups.

[0259] The term "alkoxy" refers to an -O-alkyl substituent.

[0260] As used herein, the terms “halogen,” “hal,” or “halo” mean -F, -Cl, -Br, or -I.

[0261] The term "alkylthio" refers to an -S-alkyl substituent.

[0262] The term "alkoxyalkyl" refers to an alkyl-O-alkyl substituent.

[0263] The term "haloalkoxy" refers to an -O-alkyl group substituted with one or more halo substituents. Examples of haloalkoxy groups include trifluoromethoxy and 2,2,2-trifluoroethoxy.

[0264] The term "haloalkoxyalkyl" refers to -alkyl-O-alkyl', where alkyl' is substituted by one or more halo substituents.

[0265] The term "haloalkylaminocarbonyl" refers to -C(O)-amino-alkyl, where the alkyl group is substituted by one or more halo substituents.

[0266] The term "haloalkylthio" refers to an -S-alkyl group substituted with one or more halo substituents. Examples of haloalkylthio groups include trifluoromethylthio and 2,2,2-trifluoroethylthio.

[0267] The term "haloalkylcarbonyl" refers to a -C(O)-alkyl group substituted with one or more halo substituents. An example of a haloalkylcarbonyl group is trifluoroacetyl.

[0268] The term "cycloalkyl" refers to a 3- to 8-membered monocyclic or 7- to 14-membered bicyclic ring system of hydrocarbons having at least one saturated ring or at least one non-aromatic ring (which may have some degree of unsaturation). Cycloalkyl groups may be substituted with one or more substituents as needed. In one embodiment, zero, one, two, three, or four atoms of each ring of the cycloalkyl group may be substituted with substituents. Typical examples of cycloalkyl groups include cyclopropyl, cyclopentyl, cyclohexyl, cyclobutyl, cycloheptyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, and cyclohexadienyl.

[0269] The term "cycloalkoxy" refers to an -O-cycloalkyl substituent.

[0270] The term "cycloalkoxyalkyl" refers to an alkyl-O-cycloalkyl substituent.

[0271] The term "cycloalkylalkoxy" refers to an -O-alkyl-cycloalkyl substituent.

[0272] The term "cycloalkylaminocarbonyl" refers to a -C(O)-NH-cycloalkyl substituent.

[0273] The term "aryl" refers to a monocyclic, bicyclic, or tricyclic aromatic ring system of hydrocarbons. An aryl group may be substituted with one or more substituents as needed. In one embodiment, zero, one, two, three, four, five, or six atoms in each ring of the aryl group may be substituted by substituents. Examples of aryl groups include phenyl, naphthyl, anthracenyl, fluorenyl, indenyl, and azlenyl.

[0274] The term "aryloxy" refers to an -O-aryl substituent.

[0275] The term "arylalkoxy" refers to an -O-alkyl-aryl substituent.

[0276] The term "arylalkylthio" refers to an -S-alkyl-aryl substituent.

[0277] The term "arylthioalkyl" refers to an alkyl-S-aryl substituent.

[0278] The term "arylalkylaminocarbonyl" refers to a -C(O)-amino-alkyl-aryl substituent.

[0279] The term "arylalkylsulfonyl" refers to the -S(O)2-alkyl-aryl substituent.

[0280] The term "arylalkylsulfinyl" refers to the -S(O)-alkyl-aryl substituent.

[0281] The term "aryloxyalkyl" refers to an alkyl-O-aryl substituent.

[0282] The term "alkylaryl" refers to an aryl-alkyl substituent.

[0283] The term "arylalkyl" refers to an alkyl-aryl substituent.

[0284] The term "heteroaryl" refers to an aromatic 5- to 8-membered monocyclic, 8- to 12-membered bicyclic, or 11- to 14-membered tricyclic ring system having 1 to 4 ring heteroatoms in the case of a monocyclic, 1 to 6 ring heteroatoms in the case of a bicyclic, or 1 to 9 ring heteroatoms in the case of a tricyclic, where these heteroatoms are selected from O, N, or S, and the remaining ring atoms are carbon (with appropriate hydrogen atoms unless otherwise specified). Heteroaryl groups may be substituted as needed with one or more substituents. In one embodiment, 0, 1, 2, 3, or 4 atoms in each ring of the heteroaryl group may be substituted by substituents. Examples of heteroaryl groups include pyridyl and furani. Examples include thienyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, thiazolyl, isoxazolyl, quinolinyl, pyrazolyl, isothiazolyl, pyridadinyl, pyrimidinyl, pyrazinyl, triazinyl, isoquinolinyl, and indazolyl.

[0285] The term "heteroarylalkyl" refers to an alkyl-heteroaryl substituent.

[0286] The term "heteroaryloxy" refers to an -O-heteroaryl substituent.

[0287] The term "heteroarylalkoxy" refers to an -O-alkyl-heteroaryl substituent.

[0288] The term "heteroaryloxyalkyl" refers to an alkyl-O-heteroaryl substituent.

[0289] The term "nitrogen-containing heteroaryl" refers to a heteroaryl group that has 1 to 4 ring nitrogen heteroatoms in the case of a monocyclic structure, 1 to 6 ring nitrogen heteroatoms in the case of a bicyclic structure, or 1 to 9 ring nitrogen heteroatoms in the case of a tricyclic structure.

[0290] The term "heterocycloalkyl" refers to a non-aromatic 3- to 8-membered monocyclic, 7- to 12-membered bicyclic, or 10- to 14-membered tricyclic ring system containing 1 to 3 heteroatoms in the case of a monocyclic, 1 to 6 heteroatoms in the case of a bicyclic, or 1 to 9 heteroatoms in the case of a tricyclic, where these heteroatoms are selected from O, N, S, B, P, or Si, and the non-aromatic ring system is fully saturated. Heterocycloalkyl groups may be substituted with one or more substituents as needed. In one embodiment, 0, 1, 2, 3, or 4 atoms on each ring of the heterocycloalkyl group may be substituted with substituents. Representative heterocycloalkyl groups include piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, 1,3-dioxolane, tetrahydrofuranyl, tetrahydrothienyl, and thirenyl.

[0291] The term "heterocycloalkylalkyl" refers to an alkyl-heterocycloalkyl substituent.

[0292] The term "alkylamino" refers to an amino substituent further substituted with one or two alkyl groups. The term "aminoalkyl" refers to an alkyl group further substituted with one or more amino groups. The term "hydroxyalkyl" or "hydroxylalkyl" refers to an alkyl group further substituted with one or more hydroxyl groups. The alkyl or aryl portion of alkylamino, aminoalkyl, mercaptoalkyl, hydroxyalkyl, mercaptoalkoxy, sulfonylalkyl, sulfonylaryl, alkylcarbonyl, and alkylcarbonylalkyl may be optionally substituted with one or more substituents.

[0293] The term "nucleic acid base" refers to nitrogen-containing biological compounds that form nucleosides. These include purine bases and pyrimidine bases. The five nucleic acid bases adenine (A), cytosine (C), guanine (G), thymine (T), and uracil (U) are referred to as primary or canonical nucleic acid bases. When nucleic acid bases are listed in a formula definition, this refers to the covalently bonded portion of the formula.

[0294] The term "modified nucleic acid base" refers to a derivative of a nucleic acid base. Examples of modified nucleic acid bases include, Examples include, but are not limited to, santin, hypoxanthine, 7-methylguanine, 5,6-dihydrouracil, 5-methylcytosine, 5-hydroxymethylcytosine, purine, 2,6-diaminopurine, and 6,8-diaminopurine. When modified nucleic acid bases are listed in the definition of a formula, this refers to the portion that is covalently bonded to the formula described.

[0295] Acids and bases useful in the methods described herein are known in the art. Acid catalysts are any acidic chemicals, which may be inorganic (e.g., hydrochloric acid, sulfuric acid, nitric acid, aluminum trichloride) or organic (e.g., camphor sulfonic acid, p-toluenesulfonic acid, acetic acid, ytterbium trifluoride). Acids are useful either catalytically or stoichiometrically to facilitate chemical reactions. Bases are any basic chemicals, which may be inorganic (e.g., sodium bicarbonate, potassium hydroxide) or organic (e.g., triethylamine, pyridine). Bases are useful either catalytically or stoichiometrically to facilitate chemical reactions.

[0296] An alkylating agent is any reagent capable of alkylating the functional group in question (e.g., the oxygen atom of an alcohol, the nitrogen atom of an amino group). Alkylating agents are known in the art, including in the references cited herein, and include alkyl halides (e.g., methyl iodide, benzyl bromide, or chlorides), alkyl sulfates (e.g., methyl sulfate), or combinations of other alkyl groups known in the art with leaving groups. A leaving group is any stable species that can detach from a molecule during a reaction (e.g., elimination reaction, substitution reaction), and is known in the art, including in references cited herein, and includes halides (e.g., I-, Cl-, Br-, F-), hydroxyl and alkoxy (e.g., -OMe, -Ot-Bu), acyloxyanions (e.g., -OAc, -OC(O)CF3), sulfonates (e.g., mesyl, tosyl), acetamides (e.g., -NHC(O)Me), carbamates (e.g., N(Me)C(O)Ot-Bu), phosphonates (e.g., -OP(O)(OEt)2), water, or alcohols (under protic conditions).

[0297] In certain embodiments, substituents on any group (e.g., alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, heterocycloalkyl, etc.) may be present on any atom of that group, and any group that may be substituted here (e.g., alkyl, alkenyl, alkynyl, aryl, aralkyl, heteroaryl, heteroaralkyl, cycloalkyl, heterocycloalkyl, etc.) may be optionally substituted with one or more substituents (which may be the same or different) each replacing a hydrogen atom. Examples of suitable substituents include alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, Examples include, but are not limited to, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, alkylcarbonyl, or arylamino-substituted aryl; arylalkylamino, aralkylaminocarbonyl, amide, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, imino, carboxamide, carbamide, carbamyl, thioureido, thiocyanato, sulfamide, sulfonylalkyl, sulfonylaryl, mercaptoalkoxy, N-hydroxyamidinyl, or N'-aryl, N''-hydroxyamidinyl. Specific implementations In terms of form, substituents on any group include alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, thiocarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, alkylcarbonyl, or arylamino-substituted aryl; arylalkylamino, aralkylaminocarbonyl, or amide. In certain embodiments, substituents on any group include alkyl, halogen, haloalkyl, cyano, nitro, alkoxy, hydroxyl, hydroxylalkyl, carboxyl, formyl, alkylcarbonyl, alkoxycarbonyl, alkylcarbonyloxy, thio, mercapto, mercaptoalkyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, or alkylamino.

[0298] The term “protecting group” or “protecting moiety” refers to a substituent commonly used to block or protect a particular functional group during the reaction of other functional groups on a compound, its derivatives, or its conjugates, and includes nitrogen protecting groups when bonded to a nitrogen atom, or oxygen protecting groups when bonded to an oxygen atom. Nitrogen and oxygen protecting groups are well known in the art and are described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3rd edition, John Wiley & Sons, 1999, which is incorporated herein by reference.

[0299] In certain embodiments, substituents present on the nitrogen atom are nitrogen protecting groups (also called amino protecting groups). Examples of nitrogen protecting groups include -OH and -OR. aa , -N(R cc )2, -C(=O)R aa -C(=O)N(R cc )2, -CO2R aa , -SO2R aa -C(=NR cc )R aa -C(=NR cc )OR aa -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc -SOR aa -C(=S)N(R cc )2, -C(=O)SR cc -C(=S)SR cc , C 1~10 Alkyl (e.g., aralkyl, heteroaralkyl), C 2~10 Alkenil, C 2~10 Alkinyl, C 3~10 Carbocyclyl, 3-membered to 14-membered heterocyclyl, C 6~14 Examples include, but are not limited to, aryl and 5-membered and 14-membered heteroaryl groups, where alkyl, alkenyl, alkynyl, carbocyryl, heterocyclyl, aralkyl, aryl, and heteroaryl can independently have 0, 1, 2, 3, 4, or 5 R dd It is substituted with R aa , R bb , and R cc Each of these is independently alkyl, cycloalkyl, aryl, or heteroaryl, and each of these is 1 to 3 independent R dd It can be substituted as needed, and each R ddThese are independently alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, halogen, haloalkyl, cyano, nitro, alkoxy, aryloxy, hydroxyl, hydroxylalkyl, oxo (i.e., carbonyl), carboxyl, formyl, alkylcarbonyl, alkylcarbonylalkyl, alkoxycarbonyl, alkylcarbonyloxy, aryloxycarbonyl, heteroaryloxy, heteroaryloxycarbonyl, thio, mercapto, mercaptoalkyl, arylsulfonyl, amino, aminoalkyl, dialkylamino, alkylcarbonylamino, alkylaminocarbonyl, alkoxycarbonylamino, alkylamino, arylamino, diarylamino, alkylcarbonyl, or arylamino-substituted aryl; arylalkylamino, aralkylamino The nitrogen protecting groups are rubonyl, amide, alkylaminosulfonyl, arylaminosulfonyl, dialkylaminosulfonyl, alkylsulfonylamino, arylsulfonylamino, imino, carbamide, carbamyl, thioureido, thiocyanate, sulfamide, sulfonylalkyl, sulfonylaryl, or mercaptoalkoxy. The nitrogen protecting groups are well known in the art and are incorporated herein by reference as Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3 rd Details can be found in edition, John Wiley & Sons, 1999.

[0300] Amide nitrogen protecting group (e.g., -C(=O)R) aaExamples of such derivatives include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolineamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetamide, (N'-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)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, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0301] Carbamate nitrogen protecting group (e.g., -C(=O)OR) aaExamples include methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluorenylmethyl 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 carbamate. Silylethyl (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-Bumeoc) , 2-(2'- and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamide)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl carbamate Ludithio, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, carbamate [2-(1,3-Dithianyl)methyl (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Pp, oc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzoisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, carbamate Phenyl(o-nitrophenyl)methyl, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamide)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamide) 1,1-dimethylpropynyl carbamate, 2-pyridylmethyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isobornyl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3 Examples include, but are not limited to, 5-dimethoxyphenyl)ethyl, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzyl carbamate.

[0302] Sulfonamide nitrogen protecting group (e.g., -S(=O)2R) aaExamples 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), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6- Examples include, but are not limited to, 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.

[0303] Other nitrogen 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, N-1,1,4,4-tetramethyldisilyl azacyclopentane adduct (STABASE), and 5-substituted 1,3-dimethyl-1,3,5-triazacy Chlohexane-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-pyroolin-3-yl)amine, quaternary ammonium salt, N-benzylamine, N-di(4-methoxy) Phenylmethylamine, N-5-dibenzosperylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine Np-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-hydroxyphenyl)phenylmethyleneamine, N-chlorohexylideneamine, N-(5,5-di Examples include, but are not limited to, methyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylboronic acid derivatives, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkylphosphoramide, dibenzylphosphoramide, diphenylphosphoramide, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

[0304] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also called a hydroxyl protecting group). Examples of oxygen protecting groups include -R aa , -N(R bb )2, -C(=O)SR aa -C(=O)R aa , -CO2Raa -C(=O)N(R bb )2, -C(=NR bb )R aa -C(=NR bb )OR aa -C(=NR bb )N(R bb )2, -S(=O)R aa , -SO2R aa , -Si(R aa )3, -P(R cc )2, -P(R cc )3, -P(=O)2R aa -P(=O)(R aa )2, -P(=O)(OR cc )2, -P(=O)2N(R bb )2, and -P(=O)(NR bb )2 is one example, but it is not limited to these, and here R aa , R bb , and R cc The oxygen protecting group is as defined herein. The oxygen protecting group is well known in the art and is incorporated herein by reference as Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3 rd Details can be found in edition, John Wiley & Sons, 1999.

[0305] Examples of oxygen protecting groups include methyl, methoxymethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, and 2-(trimethylsilyl)ethoxymethyl (SEMOR). ), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-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(Lebrinoyloxyphenyl)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, Benzoisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyl t-hexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoyl formate, acetate, chloroacetate, dichloroacetate, trichloroacetate Trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (rebrinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), t-butyl carbonate (BOC), alkyl methyl carbonate, 9-fluorenylmethyl Tyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphoniol)ethyl carbonate (Peoc), alkyl Sobutyl 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-nitrobenz Calcium carbonate, alkyl-S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl Tyl carbonate, dithiomethyl carbonate, 2-iodobenzoate, 4-azidobutyle 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 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-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkyl=N,N, N',N'-tetramethylphosphorodiamidate, alkyl=N-phenylcarbamate , borate, dimethylphosphine oil, alkyl=2,4-dinitrophenyl sulf Examples include, but are not limited to, ethanolates, sulfates, methanesulfonates (mesylates), benzylsulfonates, and tosylates (Ts).

[0306] In certain embodiments, substituents on the sulfur atom are sulfur protecting groups (also called thiol protecting groups). Examples of sulfur protecting groups include -R aa , -N(R bb )2, -C(=O)SR aa -C(=O)R aa , -CO2R aa -C(=O)N(R bb )2, -C(=NR bb )R aa -C(=NR bb )OR aa -C(=NR bb )N(R bb )2, -S(=O)R aa , -SO2R aa , -Si(R aa )3, -P(R cc )2, -P(R cc)3, -P(=O)2R aa -P(=O)(R aa )2, -P(=O)(OR cc )2, -P(=O)2N(R bb )2, and -P(=O)(NR bb )2 is one example, but it is not limited to these, and here R aa , R bb , and R cc The sulfur protecting group is as defined herein. The sulfur protecting group is well known in the art and is incorporated herein by reference as Protecting Groups in Organic Synthesis, TWGreene and PGMWuts, 3 rd Details can be found in edition, John Wiley & Sons, 1999.

[0307] "Michael acceptor" refers to α,β-unsaturated electrophiles, such as α,β-unsaturated carbonyl derivatives or α,β-unsaturated nitriles, but not limited to these.

[0308] Within the context of the Michael Acceptor definition, the following should be understood: “Electrophile” means a compound capable of accepting an electron pair; “α,β-unsaturated electrophile” means a class of compounds that include, but are not limited to, α,β-unsaturated carbonyl derivatives, α,β-unsaturated nitriles, α,β-unsaturated sulfones, or other vinyl derivatives substituted with strong electron-withdrawing groups such as, but not limited to, nitro groups; “α,β-unsaturated carbonyl derivative” means a class of compounds that include, but are not limited to, α,β-unsaturated ketones, quinones, or derivatives thereof, α,β-unsaturated aldehydes, α,β-unsaturated carboxylic acid derivatives such as, but are not limited to, esters, amides, substituted amides, or maleimides or derivatives thereof. Representative examples of Michael Acceptors include, but are not limited to, acrylonitrile, acrylamide, methyl acrylate, ethyl acrylate, methyl methacrylate, 2-ethylhexyl acrylate, crotonaldehyde, methyl vinyl ketone, and acrolein.

[0309] Examples of "Michael acceptors or electrophiles that can react with nucleic acids" include species that interact unfavorably with nucleic acids, such as species that may be involved in potentially genotoxic reactions with nucleic acid nucleophiles.

[0310] The reaction of a Michael acceptor with an organic thiol to form a thiol-Michael adduct is a well-documented reaction (Chem.Commun. 2005, 669-671 and the studies cited herein). This reaction has been shown to proceed in water and organic solvents. Both acidic and basic catalysts have been used to facilitate the formation of the thiol-Michael adduct and to minimize side reactions.

[0311] In one aspect, the compounds described herein are oligomeric compounds. As used herein, “oligomeric compound” means a nucleotide sequence containing about 10 to 50 nucleotides or nucleotide base pairs. In some embodiments, the oligomeric compound is paired with the coding sequence of an expressed target nucleic acid or target gene within a cell. They have nucleic acid base sequences that are at least partially complementary. In some embodiments, when these oligomeric compounds are delivered to cells expressing a gene, they can inhibit the expression of the underlying gene. Gene expression can be inhibited in vitro or in vivo. Examples of "oligomeric compounds" include, but are not limited to, oligonucleotides, single-stranded oligonucleotides, single-stranded antisense oligonucleotides, short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short hairpin RNA (shRNA), ribozymes, and interfering RNA molecules.

[0312] As used herein, “nucleic acids” (e.g., polynucleotides, oligonucleotides, polymers of nucleotides) may consist of, but are not limited to, ribonucleic acids (e.g., composed of ribonucleosides), deoxyribonucleic acids (e.g., composed of deoxyribonucleosides), modified nucleic acids (e.g., composed of modified nucleic acid bases, sugars, and / or phosphate groups), or combinations thereof. In some embodiments, nucleic acids include ribonucleic acid (RNA). In some embodiments, nucleic acids include deoxyribonucleic acid (DNA). In some embodiments, nucleic acids include modifications (e.g., modified nucleic acid bases, modified sugars, or modified phosphates).

[0313] Nucleic acids may be single-stranded or double-stranded. In some embodiments, nucleic acids are single-stranded (e.g., ssRNA, ssDNA, or ssRNA / DNA hybrids (e.g., single-stranded nucleic acids composed of both modified or unmodified ribonucleosides and modified or unmodified deoxyribonucleosides)). In some embodiments, nucleic acids are double-stranded (e.g., composed of two single-stranded nucleic acids).

[0314] In some embodiments, nucleic acids are at least 2 (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 12 The nucleotide length is 2, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, or longer. In some embodiments, the nucleic acid is at least 5 nucleotides long. In some embodiments, the nucleic acid is at least 10 nucleotides long. In some embodiments, the nucleic acid is at least 20 nucleotides long. In some embodiments, the nucleic acid is at least 30 nucleotides long. In some embodiments, the nucleic acid is at least 40 nucleotides long. In some embodiments, the nucleic acid is at least 50 nucleotides long. In some embodiments, the nucleic acid is at least 60 nucleotides long. In some embodiments, the nucleic acid is at least 70 nucleotides long. In some embodiments, the nucleic acid is at least 80 nucleotides long. In some embodiments, the nucleic acid is at least 90 nucleotides long. In some embodiments, the nucleic acid is at least 100 nucleotides long. In some embodiments, the nucleic acid is at least 150 nucleotides long.

[0315] In some embodiments, nucleic acids are 150 (for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40) ,41,42,43,44,45,46,47,48,49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85 , 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122 , 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150) are equal to or shorter than the nucleotide length. In some embodiments, the nucleic acid is equal to or shorter than 150 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 100 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 90 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 80 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 70 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 60 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 50 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 40 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 30 nucleotides. In some embodiments, the nucleic acid is equal to or shorter than 20 nucleotides.In some embodiments, the nucleic acid is equal to or shorter than 10 nucleotides in length. In some embodiments, the nucleic acid is equal to or shorter than 5 nucleotides in length.

[0316] In some embodiments, the nucleic acid is approximately 5 to 150 nucleotides long. In some embodiments, the nucleic acid is approximately 10 to 100 nucleotides long. In some embodiments, the nucleic acid is approximately 20 to 90 nucleotides long. In some embodiments, the nucleic acid is approximately 30 to 80 nucleotides long. In some embodiments, the nucleic acid is approximately 40 to 70 nucleotides long. In some embodiments, the nucleic acid is approximately 50 to 60 nucleotides long.

[0317] In some embodiments, the nucleic acid is a therapeutic nucleic acid. The therapeutic nucleic acid may include, for example, small interfering RNA (siRNA), microRNA (miRNA), ADAR mobilization molecules, ADAR targeting moieties, guide RNA, antisense nucleic acid, or a combination thereof.

[0318] The terms “microRNA” and “miRNA,” where interchangeable in this specification, refer to short (e.g., approximately 20 to 24 nucleotides in length) non-coding ribonucleic acid (RNA) molecules that are involved in the post-translational regulation of gene expression in multicellular organisms by affecting both mRNA stability and translation. miRNAs are transcribed by RNA polymerase II as a protein-coding or non-coding portion of a capped and polyadenylated primary transcript (pri-miRNA). This primary transcript is then cleaved by the Drosha ribonuclease III enzyme to produce a stem-loop precursor miRNA (pre-miRNA) approximately 70 nucleotides in length. This is further processed in the RNAi pathway. As part of this pathway, this pre-miRNA is cleaved by cytoplasmic Dicer ribonuclease to mature miRNA and antisense miRNA star (miRNA). * ) produces a product. This mature miRNA is incorporated into the RNA-induced silencing complex (RISC), which recognizes a target mRNA via incomplete base pairing with the miRNA (i.e., partial complementarity), and most commonly results in transcriptional inhibition or destabilization of this target mRNA. This mechanism is most frequently observed through the binding of the miRNA to the 3' untranslated region (UTR) of this target mRNA, which can reduce gene expression either by inhibiting translation (e.g., by blocking the approach of ribosomes for translation) or by directly causing the degradation of the transcript. The term (i.e., miRNA) may be used herein for any form of the subject miRNA (e.g., precursor, primary, and / or mature miRNA).

[0319] The terms “small interfering RNA” and “siRNA” refer, where interchangeable in this specification, to RNA molecules that exist as non-coding double-stranded RNA (dsRNA) molecules of approximately 20 to 24 nucleotides in length (roughly similar to miRNA) and are useful in RNA interference (RNAi). siRNAs are often found to have a phosphorylated 5' end and a hydroxylated 3' end, the 3' end typically having a 2-nucleotide overhang beyond the 5' end of the antiparallel strand (e.g., the complementary strand of the dsRNA molecule). siRNAs are most frequently found to interfere with the expression of specific genes and promote (e.g., facilitate, induce, initiate) mRNA degradation, thereby preventing (e.g., inhibiting, silencing, interfering with) translation, through binding to target sequences (e.g., target gene sequences) to which they are complementary. Following integration into and separation from the RISC complex, the siRNAs cleave these target mRNAs by base-pairing them (e.g., fully complementary), thereby preventing these target mRNAs from being used as translational templates. The miRNA-loaded RISC complex, which is also part of the RNAi pathway as discussed above herein, scans cytoplasmic mRNA for potential complementarity (e.g., partial complementarity).

[0320] The term “ADAR recruiting molecule,” as used herein, means a nucleic acid configured to increase the concentration of an adenosine deaminase (ADAR) enzyme acting on ribonucleic acid at a site surrounding the nucleic acid. In some embodiments, the increased concentration is compared to the concentration at a given site where the ADAR recruiting molecule is not present. In some embodiments, the ADAR recruiting molecule comprises a double strand of double-stranded RNA.

[0321] The term “ADAR-targeting moiety,” as used herein, means a nucleic acid configured to orient an ADAR molecule to a desired location (e.g., a location). As used herein, the term “orienting” means increasing the concentration of ADAR at a desired location compared to the concentration in the absence of the ADAR-targeting moiety. In some embodiments, the ADAR-targeting moiety may be configured to control the desired location by altering the sequence and / or properties of the nucleic acid (e.g., by modification of nucleic acid bases, sugars, phosphates, or other components). In some embodiments, the ADAR-targeting moiety includes an ADAR mobilizing molecule and a single-stranded guide nucleic acid. In some embodiments, the ADAR-targeting moiety includes a double-stranded RNA and a single-stranded guide nucleic acid.

[0322] The term “single-stranded guide nucleic acid” means, as may be used herein, a single-stranded nucleic acid comprising a specific sequence that is at least partially complementary to the target sequence. In some embodiments, this target sequence is located at, adjacent to, or near a desired location for regulating ADAR concentration. In some embodiments, the level of complementarity is sufficient to facilitate the binding (e.g., annealing) of the single-stranded guide nucleic acid to the target sequence.

[0323] The term “antisense molecule,” as used herein, means an oligonucleotide (e.g., a polymer of nucleotides) that synthesizes or contains a sequence of nucleotides complementary to a target nucleic acid sequence. For example, with respect to RNA (e.g., mRNA, miRNA), the strand may be read as 5'-AAGGUCCU-3', while the antisense molecule is read as 3'-UUCCAGGA-5'. In the case of antisense molecules targeting RNA, they can regulate expression in various ways. For example, the strand may target mRNA (thus blocking translation and promoting the degradation of this mRNA transcript), or in other ways, these strands may target miRNA (thus inhibiting the targeting of the blocked miRNA by mRNA, and inhibiting the promotion or recovery of translation from this mRNA, and promoting the degradation of this blocked miRNA).

[0324] In some embodiments, nucleic acids bind to the GalNAc moiety. GalNAc (N-acetylgalactosamine) is an amino sugar derivative of galactose. In some embodiments, the GalNAc moiety is structured [ka] Includes. In some embodiments, the GalNAc portion is structure [ka] The GalNAc moiety is a portion that has affinity for receptors in various tissues and cells. In this way, the GalNAc moiety can facilitate the targeting of cargo (e.g., nucleic acids) to such tissues and receptors. In some embodiments, the GalNAc moiety is useful for directing nucleic acids. In some embodiments, the GalNAc moiety directs nucleic acids to a certain location. In some embodiments, the GalNAc moiety targets a tissue. In some embodiments, this tissue is the liver. In some embodiments, the GalNAc moiety targets a cell receptor. In some embodiments, the cell receptor is an asialoglycoprotein receptor. In some embodiments, it is an asialoglycoprotein receptor on hepatocytes.

[0325] In some embodiments, the nucleic acid binds to one or more GalNAc moieties. In some embodiments, the nucleic acid binds to at least two GalNAc moieties (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more). In some embodiments, the nucleic acid binds to at least three GalNAc moieties. In some embodiments, the nucleic acid binds to at least five GalNAc moieties. In some embodiments, the nucleic acid binds to at least one to about ten GalNAc moieties. In some embodiments, the nucleic acid binds to at least one to about eight GalNAc moieties. In some embodiments, the nucleic acid binds to at least one to about six GalNAc moieties. In some embodiments, the nucleic acid binds to at least one to about four GalNAc moieties. In some embodiments, the nucleic acid binds to at least one to about two GalNAc moieties. In some embodiments, the nucleic acid binds to at least one to ten GalNAc moieties. Binding. In some embodiments, the nucleic acid binds to at least 1 to 8 GalNAc moieties. In some embodiments, the nucleic acid binds to at least 1 to 6 GalNAc moieties. In some embodiments, the nucleic acid binds to at least 1 to 4 GalNAc moieties. In some embodiments, the nucleic acid binds to at least 1 to 2 GalNAc moieties.

[0326] In some embodiments, the nucleic acid binds to the GalNAc moiety via either its 5' end and / or 3' end (e.g., connects, adheres, associates). In some embodiments, the nucleic acid binds to the GalNAc moiety via its 5' end. In some embodiments, the nucleic acid binds to the GalNAc moiety via its 3' end. In some embodiments, the nucleic acid binds to the GalNAc moiety via both its 5' and 3' ends. In some embodiments, the nucleic acid binds to the GalNAc moiety via its inner portion.

[0327] In several respects, this disclosure relates to methods for preparing compositions comprising nucleic acids and GalNAc, as disclosed herein.

[0328] In another aspect, the present invention provides conjugates of the formulas of the compounds herein. In one embodiment, these conjugates include a portion of the formula of the present invention covalently bonded to a portion that is a protein, nucleic acid, small molecule, large molecule, therapeutic agent, diagnostic agent, imaging agent, or targeting agent.

[0329] The conjugates disclosed herein may be manufactured using any available method. Where relating to a part of a compound formula having a drug part (e.g., a protein, nucleic acid, small molecule, large molecule, therapeutic agent, diagnostic agent, imaging agent, or targeting agent), these parts may be linked directly or indirectly (e.g., via a linker part; i.e., this linker is covalently bonded to each of the parts of the compound formula and the drug part; in some formulas herein, "-L-"). For example, the compound formula and the drug may be related to each other directly (e.g., by one or more covalent bonds) or by one or more linkers.

[0330] "Therapeutic agents" may be therapeutic agents for regulating hepatocytes or for treating liver diseases, such as viral hepatitis, hepatic fibrosis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), rare diseases, metabolic diseases, obesity, cardiovascular diseases, hemochromatosis, thalassemia, hepatic trauma, alcoholic liver disease, or hereditary liver disorders, and may be oligonucleotides, small molecules, or peptides, such as ursodiol, meticlothiazide, pioglitazone, metadoxine, cystadane, ondansetron, silymarin, lamivudine, adefovir, tenofovir disoproxil, tenofovir alafenamide, terbivudine, entecavir, and cholic acid.

[0331] Any suitable linker may be used in accordance with the present invention. The linker may be used to form amide bonds, ester bonds, disulfide bonds, etc. The linker may contain carbon atoms or heteroatoms (e.g., nitrogen, oxygen, sulfur, etc.). Typically, the linker may be 1 to 50 atoms long, 1 to 40 atoms long, 1 to 25 atoms long, 1 to 20 atoms long, 1 to 15 atoms long, 1 to 10 atoms long, or 1 to 5 atoms long. The linker may be substituted with various substituents, including but not limited to hydrogen atoms, alkyl, alkenyl, alkynyl, amino, alkylamino, dialkylamino, trialkylamino, hydroxyl, alkoxy, halogen, aryl, heterocyclic, aromatic heterocyclic, cyano, amide, carbamoyl, carboxylic acid, ester, thioether, alkylthioether, thiol, and ureido groups. To ensure this understanding, each of these groups may be further substituted. This linker may be an aliphatic linker or a heteroaliphatic linker. For example, this linker may be a polyalkyl linker. This linker may be a polyether linker. This linker may be a polyethylene linker, for example, PEG. This linker may be a short peptide chain, for example, 1 to 10 amino acid lengths, for example, 1, 2, 3, 4, or 5 amino acid lengths, nucleic acids, alkyl chains, etc.

[0332] The compounds and oligonucleotides of this disclosure can be prepared by means known in the art of organic synthesis. Methods for optimizing reaction conditions and, if necessary, minimizing competing by-products are known in the art. Reaction optimization and scale-up can be advantageously performed using high-speed parallel synthesis facilities and computer-controlled microreactors (e.g., Design and Optimization in Organic Synthesis, 2). nd Edition,Carlson R,Ed,2005;Elsevier Science Ltd.;Jaehnisch,K et al. Angew. Chem. Int. Ed. Engl. 2004 43:406; and references within these). Further reaction schemes and protocols may be determined by those skilled in the art using commercially available structure-searchable database software, such as SciFinder® (CAS Division of the American Chemical Society) and CrossFire Beilstein® (Elsevier MDL), or by appropriate keyword searches using internet search engines such as Google® or keyword databases such as the U.S. Patent and Trademark Office text database.

[0333] As can be understood by those skilled in the art, the methods for synthesizing the compounds and oligonucleotides of the formulas herein, including the schemes and examples herein, are obvious to those skilled in the art. Accordingly, various synthesis steps may be carried out in alternative sequences or orders to obtain the desired compounds and oligonucleotides. Furthermore, the solvents, temperatures, reaction times, etc., described herein are for illustrative purposes only, and those skilled in the art will recognize that variations in reaction conditions may produce the desired compounds and oligonucleotides of this disclosure.

[0334] The compounds and oligonucleotides herein may also include bonds (e.g., carbon-carbon bonds) where the rotation of the bond is restricted around this particular bond, for example, by the presence of a ring or double bond. Thus, all cis / trans and E / Z isomers are expressly included in this disclosure. The compounds and oligonucleotides herein may also be represented in multiple tautomers, and in such examples, this disclosure expressly encompasses all tautomers of the compounds and oligonucleotides described herein, even if only a single tautomer is represented. All such isomeric forms of the compounds and oligonucleotides herein are expressly included in this disclosure. All crystalline forms and polymorphisms of the compounds and oligonucleotides described herein are expressly included in this disclosure. Extracts and fractions containing the compounds and oligonucleotides of this disclosure are also practiced. The term “isomer” is intended to encompass diastereoisomers, enantiomers, positional isomers, structural isomers, rotational isomers, and tautomers, etc. With respect to compounds and oligonucleotides containing one or more stereogenic centers, such as chiral compounds, the methods of this disclosure may be carried out using enantiomer-enriched compounds, racemates, or mixtures of diastereomers. All isomers of the compounds described herein are expressly incorporated herein.

[0335] Preferred enantiomer-enriched compounds have an enantiomer excess of 50% or more, and more preferably, the compound has an enantiomer excess of 60%, 70%, 80%, 90%, 95%, 98%, or 99% or more. In the application method, only one enantiomer or diastereomer of the chiral compound of the present disclosure is administered to cells or subjects.

[0336] Treatment method A method is provided for treating a subject who is suffering from or susceptible to a disability or disease, the method comprising administering to the subject an effective amount of a compound, oligonucleotide, or pharmaceutical composition of any of the formulas herein (e.g., formulas I to XXXIV).

[0337] In another context, a method is provided for treating a subject who is suffering from or susceptible to a disorder or disease, and the subject is identified as needing modification of protein function, and the method comprises the step of administering to the subject in need of this treatment an effective amount of a compound, oligonucleotide, or pharmaceutical composition of any of the formulas herein (e.g., formulas I to XXXIV), as a result the subject is treated for the disorder.

[0338] In one aspect, a method is provided for modulating protein function in a subject, the method comprising contacting the subject with a compound or oligonucleotide of any of the formulas herein (e.g., formulas I to XXXIV) in a sufficient amount and under conditions to modulate protein function.

[0339] In one embodiment, this regulation is inhibition.

[0340] In some embodiments, a method is provided for treating liver cells in a subject, the method comprising administering to a subject requiring treatment of liver cells an effective amount of any compound, oligonucleotide, or pharmaceutical composition of any of the formulas herein (e.g., formulas I to XXXIV) in an amount and under conditions sufficient to target the liver cells.

[0341] In certain embodiments, methods are provided for treating a disease, disorder, or symptoms thereof, wherein the disorder is cancer, proliferative disorders, neurodegenerative diseases, autoimmune or inflammatory disorders, infections, metabolic disorders, hematological disorders, or cardiovascular diseases.

[0342] In certain embodiments, the disorder or disease is cancer or a proliferative disorder. In certain embodiments, this cancer or proliferative disorder includes carcinoma, leukemia, blastoma, lymphoma, myeloma, or melanoma, or a combination thereof. In certain embodiments, the disorder or disease is multiple myeloma, melanoma, breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, hepatocellular carcinoma, kidney cancer, leukemia, T-cell lymphoma, bone cancer, gliablastoma, neuroblastoma, oral squamous cell carcinoma, urothelial carcinoma, lung cancer, cervical cancer, colon cancer, head and neck squamous cell carcinoma, Burkitt lymphoma, esophageal cancer, Hodgkin lymphoma, bladder cancer, or stomach cancer, or a combination thereof.

[0343] In certain embodiments, the disorder or disease is rheumatoid arthritis, spondyloarthritis, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, graft-versus-host disease, transplant rejection, fibrous disease, Crohn's disease, type 1 diabetes, eczema, psoriasis, sepsis, airway hyperresponsiveness, ulcerative colitis, or a combination thereof.

[0344] In certain embodiments, the injury or disease is epilepsy, attention deficit disorder, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal muscular atrophy, essential tremor, central nervous system trauma, multiple sclerosis, Charcot-Marie-Tooth (MCT), This could be peripheral neuropathy, cerebral ischemia, or a combination of both.

[0345] In certain embodiments, the disorder or disease is an infection caused by a virus, fungus, or bacteria, or a combination thereof.

[0346] In certain embodiments, the disorder or disease is metabolic syndrome, diabetes mellitus, obesity, hypertension, heart failure, cystic growth in autosomal dominant polycystic kidney disease (ADPKD), or a combination thereof.

[0347] In certain embodiments, the disorder or disease is cardiovascular stress, pressure overload, chronic ischemia, infarction-reperfusion injury, hypertension, atherosclerosis, peripheral artery disease, heart failure, hypertrophy, angina, arrhythmia, hypercholesterolemia, atherosclerosis, or seizure, or a combination thereof.

[0348] In certain embodiments, the disorder or disease is a liver disease.

[0349] In certain embodiments, the subject is a mammal, preferably a primate or a human.

[0350] In another embodiment, the method described above is provided, wherein the effective amount of any compound or oligonucleotide of any of the formulas (e.g., formulas I to XXXIV) described herein is as described above.

[0351] In another embodiment, the method described above is provided, wherein a compound or oligonucleotide of any of the formulas herein (e.g., formulas I to XXXIV) is administered intravenously, intramuscularly, subcutaneously, intraventricularly, orally, or topically.

[0352] In other embodiments, the methods described above are provided, in which any compound or oligonucleotide of any of the formulas herein (e.g., formulas I to XXXIV) is administered alone or in combination with one or more other therapeutic agents. In further embodiments, these additional therapeutic agents are anticancer agents, antifungal agents, cardiovascular agents, anti-inflammatory agents, chemotherapeutic agents, anti-angiogenic agents, cytotoxic agents, antiproliferative agents, metabolic agents, ophthalmologic agents, central nervous system (CNS) agents, urinary tract agents, or gastrointestinal agents.

[0353] Another purpose of this disclosure is the use of the compounds or oligonucleotides described herein (e.g., compounds or oligonucleotides of formulas I to XXXIV) in the manufacture of pharmaceuticals for use in the treatment of disorders or diseases. Another purpose of this disclosure is the use of the compounds or oligonucleotides described herein (e.g., compounds or oligonucleotides of formulas I to XXXIV) for use in the treatment of disorders or diseases. Another purpose of this disclosure is the use of the compounds or oligonucleotides described herein (e.g., compounds or oligonucleotides of formulas I to XXXIV) in the manufacture of pesticide compositions for use in the treatment or prevention of disorders or diseases in agricultural or agrarian environments.

[0354] Pharmaceutical composition In one aspect, a pharmaceutical composition is provided that contains a compound or oligonucleotide of any of the formulas herein (e.g., formulas I to XXXIV) and a pharmaceutically acceptable carrier.

[0355] The compounds, oligonucleotides, or compositions described herein are one or They may be administered in combination with more additional therapeutic agents (e.g., therapeutic and / or prophylactically active agents). These compounds, oligonucleotides, or compositions may be administered in combination with additional therapeutic agents that improve their activity (e.g., activity to treat a disease in a subject that requires treatment of the disease, to prevent a disease in a subject that requires prevention of the disease, and / or activity to reduce the risk of a subject developing a disease in a subject that requires reduction of the risk of developing the disease (e.g., efficacy and / or potency)), improve bioavailability, improve safety, reduce drug resistance, reduce and / or modify metabolism, inhibit elimination, and / or modify distribution in a subject or cell. It is also understood that the treatments employed may achieve the desired effect with respect to the same disorder, and / or may achieve different effects. In certain embodiments, the pharmaceutical compositions described herein contain the compounds or oligonucleotides described herein and additional therapeutic agents to exhibit synergistic effects not present in pharmaceutical compositions that contain either the compound or oligonucleotide and the additional therapeutic agent, but not both.

[0356] Compounds, oligonucleotides, or combinations may be administered simultaneously with, before, or after, one or more additional therapeutic agents, which may be useful, for example, as combination therapy. Therapeutic agents contain therapeutically active agents. Therapeutic agents also contain prophylactically active agents. Therapeutic agents include organic small molecules such as drug compounds (e.g., compounds approved for use in human and veterinary medicine by the U.S. Food and Drug Administration as given in the Code of Federal Regulations (CFR)), peptides, proteins, carbohydrates, monosaccharides, oligosaccharides, polysaccharides, nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides or proteins, protein-bound small molecules, glycoproteins, steroids, nucleic acids, DNA, RNA, nucleotides, nucleosides, oligonucleotides, antisense oligonucleotides, lipids, hormones, vitamins, and cells. In certain embodiments, additional therapeutic agents are therapeutic agents useful for treating and / or preventing diseases (e.g., cancer, proliferative disorders, neurodegenerative diseases, autoimmune or inflammatory disorders, infections, metabolic disorders, hematological disorders, cardiovascular diseases). Each additional therapeutic agent may be administered in a dose and / or time schedule determined for that therapeutic agent. These additional therapeutic agents may also be administered together with each other and / or with the compounds or compositions described herein, in a single dose or separately in different doses. Specific combinations to be adopted in a regimen should take into consideration the compatibility of the compounds described herein with the additional therapeutic agent(s), and / or the therapeutic and / or preventive effects to be achieved. In general, the additional therapeutic agent(s) in a combination are expected to be used at levels not exceeding those used individually. In some embodiments, the levels used in combination are lower than those used individually.

[0357] Additional therapeutic agents include, but are not limited to, antiproliferative agents, anticancer agents, anti-angiogenic agents, anti-inflammatory agents, and immunosuppressants. In certain embodiments, the additional therapeutic agent is immunotherapy. In certain embodiments, the additional therapeutic agent is an antiproliferative agent. In certain embodiments, the additional therapeutic agent is an anticancer agent. In certain embodiments, anticancer agents include epigenetic or transcription modulators (e.g., DNA methyltransferase inhibitors, histone deacetylase inhibitors (HDAC inhibitors), lysine methyltransferase inhibitors), antimitotic agents (e.g., taxanes and vinca alkaloids), cell signaling pathway inhibitors (e.g., tyrosine protein kinase inhibitors), protein stability modulators (e.g., proteasome inhibitors), Hsp90 inhibitors, glucocorticoids, all-trans retinoic acids, antiestrogens (e.g., tamoxifen, raloxifene, and megestrol), LHRH agonists (e.g., goscrclin and leuprolide), antiandrogens (e.g., flutamide and bicalutamide), and photodynamic therapies (e.g., vertporfin). oporfin (BPD-MA), phthalocyanines, photosensitizer Pc4, and demethoxy-hypocrelin A (2BA-2-DMHA), nitrogen mustards (e.g., cyclophosphamide, ifosfamide, trophosfamide, chlorambucil, estramustine, and melphalan), nitrosoureas (e.g., carmustine (BCNU) and lomustine (CCNU)), alkyl sulfonates (e.g., busulfan and treosulfan), triazenes (e.g., dacarbazine, temozolomide), platinum-containing compounds (e.g., cisplatin, carboplatin, oxaliplatin) (e.g., vincristine, vinblastine, vindesine, and vinorelbine), taxoids (e.g., paclitaxel or paclitaxel equivalents, e.g., nanoparticle albumin-conjugated paclitaxel (Abraxane), docosahexanoic acid-conjugated paclitaxel (DHA-paclitaxel, taxoprexin), polyglutamate-conjugated paclitaxel (PG-paclitaxel, paclitaxel polyglutex) (Poliglumex), CT-2103, XYOTAX), tumor-activating prodrug (TAP) ANG1005 (Angiopep-2 bound to 3 molecules of paclitaxel), paclitaxel-EC-1 (paclitaxel bound to erbB2-recognizing peptide EC-1), and glucose-bound paclitaxel, e.g., 2'-paclitaxel=methyl=2-glucopyranosyl=succinate; docetaxel, taxol), epipodophilin (epipodo phyllins) (e.g., etoposide, etoposide phosphate, teniposide, topotecan, 9-aminocamptothecin, camptoirinotecan, irinotecan, cristol, mitomycin C) C)), antimetabolites, DHFR inhibitors (e.g., methotrexate, dichloromethotrexate, trimethrexate, edatrexate), IMP dehydrogenase inhibitors (e.g., mycophenolic acid, tiazofurin, ribavirin, and EICAR), ribonucleotide reductase inhibitors (e.g., hydroxyurea and deferoxamine), uracil analogs (e.g., 5-fluorouracil (5-FU), phloxuridine, doxifluridine, ratitrexed, tegafur-uracil, capecitabine), cytosine analogs (e.g., cytarabine (ara C), cytosine arabinoside, and fludarabine), purine analogs (e.g., mercaptopurine and thioguanine), vitamin D3 analogs (e.g., EB 1089, CB) 1093, and KH 1060), isoprenement inhibitors (e.g., lovastatin), dopaminergic neurotoxins (e.g., 1-methyl-4-phenylpyridinium ion), cell cycle inhibitors (e.g., staurosporine), actinomycin (e.g., actinomycin D, dactinomycin), bleomycin (e.g., bleomycin A2, bleomycin B2, peplomycin), anthracyclines (e.g., daunorubicin, doxorubicin, pegylated liposomal doxorubicin, idarubicin, epirubicin, pirarubicin, zorubicin, mitoxantrone), MDR inhibitors (e.g., verapamil), Ca2+ ATPase inhibitors (e.g., thapsigardin), thalidomide, lenalidomide, pomalidomide, tyrosine kinase inhibitors (e.g., axitinib (AG013736), bosutinib (SKI-606), cediranib (RECENTINTM, AZD2171), dasatinib (SPRYCEL®, BMS-354825), erlotinib (TARCEVA®), gefitinib (Iressa®), imatinib (Gleevec®, CGP57148B, STI-571), lapatinib (TYKERB®, TYKERB®) VERB (registered trademark), Restaurtinib (CEP-701), Neratinib (HKI-272), Nilotinib (TASIGNA (registered trademark)), Semaxanib (semaxinib, SU5416), Sunitinib (SUTENT (registered trademark), SU11248), Toceranib (PALLADIA (registered trademark)), Vandetanib (ZACTIMA (registered trademark), ZD6474), Vatalanib (PTK787, PTK / ZK), Trastuzumab (Herceptin (registered trademark) Trademarks)), bevacizumab (AVASTIN®), rituximab (Rituxan®), cetuximab (Erbitux®), panitumumab (VECTIBIX®), ranibizumab (Lucentis®), nilotinib (TASIGNA®), sorafenib (NEXAVAR®), everolimus (AFINITOR®), alemtuzumab (CAMPATH®), gemtuzumab ozogamicin (Mylotarg®), temsirolimus (TORISEL®), ENMD-2076, PCI-32765, AC220, dovitinib lactate (TKI258, CHIR-258), BIBW 2992 (TOVOK™), SGX523, PF-04217903, PF-02341066, PF-299804, BMS-777607, ABT-869, MP470, BIBF 1120 (VARGATEF®), AP24534, JNJ-26483327, MGCD265, DCC-2036, BMS-690154, CEP-11981, tivozanib (AV-951), OSI-930, MM-121, XL-184, XL-647, and / or XL228), proteasome inhibitors (e.g., bortezomib (Velkk) (e.g., ixazomib (NINLARO)), mTOR inhibitors (e.g., rapamycin, temsirolimus (CCI-779), everolimus (RAD-001), ridafololimus, AP23573 (Ariad), AZD8055 (AstraZeneca), BEZ235 (Novartis), BGT226 (Norvartis), XL765 (Sanofi)Examples include, but are not limited to, Aventis, PF-4691502 (Pfizer), GDC0980 (Genentech), SF1126 (Semafoe), and OSI-027 (OSI), oblimersen, gemcitabine, carminomycin, leucovorin, pemetrexed, cyclophosphamide, dacarbazine, procarbazine, prednisolone, dexamethasone, campathecin, plicamycin, asparaginase, aminopterin, metopterin, porphyromycin, melphalan, leurosidine, leurosine, chlorambucil, trabectedin, procarbazine, discodermoldide, carminomycin, aminopterin, and hexamethylmelamine.

[0358] In certain embodiments, the additional therapeutic agent is immunotherapy. In certain embodiments, this immunotherapy is useful in the treatment of cancer. Exemplary immunotherapies include, but are not limited to, T-cell therapy, interferon, cytokines (e.g., tumor necrosis factor, interferon-α, interferon-γ), vaccines, hematopoietic growth factors, monoclonal serum therapy, immunoenhancing and / or immunomodulatory agents (e.g., IL-1, 2, 4, 6, or 12), immune cell growth factors (e.g., GM-CSF), and antibodies. In certain embodiments, the immunotherapy is T-cell therapy. In certain embodiments, this T-cell therapy is chimeric antigen receptor T cells (CAR-T). In certain embodiments, the immunotherapy is an antibody. In certain embodiments, this antibody is an anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4 antibody, anti-TIM3 antibody, anti-OX40 antibody, anti-GITR antibody, anti-LAG-3 antibody, anti-CD137 antibody, anti-CD27 antibody, anti-CD28 antibody, anti-CD28H antibody, anti-CD30 antibody, anti-CD39 antibody, anti-CD40 antibody, anti-CD47 antibody, anti-CD48 antibody, anti-CD70 antibody, anti-CD73 antibody, anti-CD96 antibody, anti-CD160 antibody, anti-CD200 antibody, anti-CD244 antibody, anti-ICOS antibody, anti-TNFRSF25 antibody , anti-TMIGD2 antibody, anti-DNAM1 antibody, anti-BTLA antibody, anti-LIGHT antibody, anti-TIGIT antibody, anti-VISTA antibody, anti-HVEM antibody, anti-Siglec antibody, anti-GAL1 antibody, anti-GAL3 antibody, anti-GAL9 antibody, anti-BTNL2 (butyrophilin (butrophylins)) antibody, anti-B7-H3 antibody, anti-B7-H4 antibody, anti-B7-H5 antibody, anti-B7-H6 antibody, anti-KIR antibody, anti-LIR antibody, anti-ILT antibody, anti-MICA antibody, anti-MICB antibody, anti-NKG2D antibody, anti-NKG2A antibody, anti-T GFβ antibody, anti-TGFβR antibody, anti-CXCR4 antibody, anti-CXCL12 antibody, anti-CCL2 antibody, anti-IL-10 antibody, anti-IL-13 antibody, anti-IL-23 antibody, anti-hosphemyline antibody, anti-neuropilin antibody, anti-GalCer antibody, anti-HER2 antibody, anti-VEGFA antibody, anti-VEGFR antibody, anti-EGFR antibody, またはanti-Tie2 antibodyである.In certain embodiments, this antibody is used to target pembrolizumab, nivolumab, pidilizumab, ipilimumab, tremelimumab, durvalumab, atezolizumab, avelumab, PF-06801591, utomilumab, PDR001, PBF-509, MGB453, LAG525, AMP-224, INCSHR1210, INCAGN1876, INCAGN1949, samalizumab, PF-05082566, urelumab, lirilumab, lulizumab, BMS-936559, BMS-936561, BMS-986004, BMS-986012, BMS-986016, BMS-986178, IMP321, IPH2101, IPH2201, varilumab, ulocuplumab, monalizumab, MEDI0562, MEDI0680, MEDI1873, MEDI6383, MEDI646 9, MEDI9447, AMG228, AMG820, CC-90002, CDX-1127, CGEN15001T, CGEN15022, CGEN15029, CGEN15049, CGEN15027, CGEN15052, CGEN15092, CX-072, CX-2009, CP-870893, Lucatumumab, Dacetuzumab, Chi These include Lob 7 / 4, RG6058, RG7686, RG7876, RG7888, TRX518, MK-4166, MGA271, IMC-CS4, emactuzumab, trastuzumab, pertuzumab, obinutuzumab, cabiralizumab, margetuximab, enoblituzumab, mogamulizumab, panitumumab, carlumab, bevacizumab, rituximab, or cetuximab.

[0359] In certain embodiments, the compounds, oligonucleotides, or pharmaceutical compositions described herein may be administered in combination with anticancer therapies, including but not limited to surgery, radiotherapy, and transplantation (e.g., stem cell transplantation, bone marrow transplantation).

[0360] In certain embodiments, additional therapeutic agents are selected from the group consisting of Akt inhibitors, alkylating agents, androgen receptor antagonists, anti-estrogens, Bcl-2 inhibitors, BRAF kinase inhibitors, BTK inhibitors, CAR-T cells, anti-CD38 antibodies, CDK inhibitors, anti-CTLA-4 antibodies, ERK / MAPK inhibitors, farnesyltransferase inhibitors, IL-6 inhibitors, immunomodulators, oncological immunotherapy agents, JAK2 / FLT3 inhibitors, kinesin spindle protein inhibitors, MEK inhibitors, anti-PD-1 antibodies, anti-PD-L1 antibodies, PI3K inhibitors, proteasome inhibitors, radioactive materials (sensitizers), radioisotopes (sensitizers), synthetic retinoids (AM80), taxanes, tyrosine kinase inhibitors, VDR agonists, VEGF inhibitors, oncolytic viruses, and combinations thereof. In certain embodiments, additional therapeutic agents include all-trans retinoic acid (ATRA), arsenic trioxide, berberine, bevacizumab, bortezomib, cabazitaxel, carfilzomib, cisplatin, clarithromycin, cyclophosphamide, cytarabine, darazalex, dexamethasone, docetaxel, elotuzumab, enzalutamide, epirubicin, fluorouracil (5-FU), gefitinib, gemcitabine hydrochloride, ibrutinib, idelalisib, indatuximab, ixazomib, ravtansine, ipilimumab, lenalidomide, lonafarnib, methotrexate, and nab-P. The drug is selected from the group consisting of clitaxel, nivolumab, paclitaxel, pacritinib, pomalidomide, sorafenib, temozolomide, thalidomide, vemurafenib, and vincristine.

[0361] In one embodiment, a kit is provided comprising, in unit dosage form, an effective amount of any compound or oligonucleotide of any of the formulas herein (e.g., formulas I to XXXIV), along with instructions for administering the compound or oligonucleotide to a subject suffering from or susceptible to any of the following diseases or disorders: cancer, proliferative disorders, neurodegenerative diseases, autoimmune or inflammatory disorders, infections, metabolic disorders, hematological disorders, and cardiovascular diseases. In another embodiment, the disease, disorder, or symptom thereof is carcinoma, leukemia, blastoma, lymphoma, myeloma, or melanoma. In other embodiments, this disease, disorder, or symptom is multiple myeloma, melanoma, breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, hepatocellular carcinoma, kidney cancer, leukemia, T-cell lymphoma, bone cancer, gliablastoma, neuroblastoma, oral squamous cell carcinoma, urothelial carcinoma, lung cancer, cervical cancer, colon cancer, head and neck squamous cell carcinoma, Burkitt lymphoma, esophageal cancer, Hodgkin lymphoma, bladder cancer, or gastric cancer. In other embodiments, this disease, disorder, or symptom is rheumatoid arthritis, spondyloarthritis, psoriatic arthritis, multiple sclerosis, systemic lupus erythematosus, inflammatory bowel disease, graft-versus-host disease, transplant rejection, fibrous disease, Crohn's disease, type 1 diabetes, eczema, psoriasis, sepsis, airway hyperresponsiveness, or ulcerative colitis. In other embodiments, this disease, disorder, or symptom is epilepsy, attention deficit disorder, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinal muscular atrophy, essential tremor, central nervous system trauma, multiple sclerosis, Charcot-Marie-Tooth (MCT), peripheral neuropathy, or cerebral ischemia. In other embodiments, this disease, disorder, or symptom is an infection caused by a virus, fungus, or bacteria. In other embodiments, this disease, disorder, or symptom is metabolic syndrome, diabetes mellitus, obesity, hypertension, heart failure, or cystic growth in autosomal dominant polycystic kidney disease (ADPKD).In other embodiments, this disease, disorder, or symptom thereof is cardiovascular stress, pressure overload, chronic ischemia, infarction-reperfusion injury, hypertension, atherosclerosis, peripheral artery disease, heart failure, hypertrophy, angina, arrhythmia, hypercholesterolemia, atherosclerosis, or seizure.

[0362] The terms “pharmaceutically acceptable salt” or “pharmaceutically acceptable carrier” encompass salts of active compounds prepared using relatively non-toxic acids or bases, depending on the specific substituents found in the compounds described herein. If the compounds of this disclosure contain relatively acidic functional groups, base addition salts may be obtained by contacting such a compound in its neutral form with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic aminos, or magnesium, or similar salts. If the compounds of this disclosure contain relatively basic functional groups, acid addition salts may be obtained by contacting such a compound in its neutral form with a sufficient amount of the desired acid, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monocarbonate, phosphoric acid, monohydrogen-phosphoric acid, dihydrogen-phosphoric acid, sulfuric acid, monohydrogen-sulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, and methanesulfonic acid. Salts of amino acids such as alginic acid, and salts of organic acids such as glucuronic acid or galacturonic acid are also included (see, for example, Berge et al., Journal of Pharmaceutical Science 66:1-19 (1977)). The compound contains both basic and acidic functional groups, thereby allowing these compounds to be converted into either a base addition salt or an acid addition salt. Other pharmaceutically acceptable carriers known to those skilled in the art are suitable for this disclosure.

[0363] Neutral forms of compounds and oligonucleotides can be regenerated by contacting their salts with a base or acid and then isolating the parent compound or oligonucleotide in a conventional manner. The parent forms of compounds or oligonucleotides differ from their various salt forms in certain physical properties, such as solubility in polar solvents, but in other respects, these salts are equivalent to the parent forms of the compounds for the purposes of this disclosure.

[0364] In addition to salt forms, this disclosure provides compounds in prodrug forms. The prodrugs of the compounds described herein are compounds that readily undergo chemical changes under physiological conditions to provide the compounds of this disclosure. Furthermore, the prodrugs can be converted to the compounds of this disclosure by chemical or biochemical methods in an ex vivo environment. For example, a prodrug can be slowly converted to the compounds of this disclosure when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0365] Certain compounds in this disclosure may exist in non-solvated forms and in solvated forms, including hydrated forms. Generally, solvated forms are equivalent to non-solvated forms and are intended to be included within the scope of this disclosure. Certain compounds in this disclosure may exist in multiple crystalline or amorphous forms. Generally, all physical forms are equivalent with respect to the uses envisioned by this disclosure and are intended to be within the scope of this disclosure.

[0366] This disclosure also provides a pharmaceutical composition containing an effective amount of the compounds described herein and a pharmaceutically acceptable carrier. In one embodiment, any compound or oligonucleotide of any of the formulas described herein (e.g., formulas I to XXXIV) is administered to a subject using a pharmaceutically acceptable formulation, for example, a sustained-release of the compound, or a pharmaceutically acceptable formulation provided for at least 12 hours, 24 hours, 36 hours, 48 ​​hours, 1 week, 2 weeks, 3 weeks, or 4 weeks after the administration of the pharmaceutically acceptable formulation to the subject.

[0367] The actual dosage levels and administration timelines of the active ingredients in the pharmaceutical compositions of this disclosure may be modified to obtain an effective amount of the active ingredient to achieve the desired therapeutic response with respect to a particular patient, composition, and route of administration, without being toxic (or unacceptably toxic) to that patient.

[0368] In use, at least one compound or oligonucleotide according to the Disclosure is administered in a pharmaceutically effective amount to a subject in need, in a pharmaceutically effective amount, by intravenous, intramuscular, subcutaneous, or intracerebroventricular injection in a pharmaceutically effective carrier, or by oral administration or topical application. According to the Disclosure, the compounds or oligonucleotides of the Disclosure may be administered alone or together with a second different therapeutic agent. "Together" means together, substantially simultaneously, or sequentially. In one embodiment, the compounds or oligonucleotides of the Disclosure are administered acutely. Thus, the compounds or oligonucleotides of the Disclosure may be administered over a short period of treatment, for example, over about one day to about one week. In another embodiment, the compounds or oligonucleotides of the Disclosure may be administered over a longer period, for example, over about one week to several months, depending on the condition to be treated, to improve a chronic disorder.

[0369] "Pharmacologically effective dose" means, as used herein, a dose of the compound or oligonucleotide disclosed that is sufficiently high to significantly and positively modify the condition to be treated, but within reasonable medical judgment, sufficiently low to avoid serious side effects (at a reasonable benefit / risk ratio). This refers to the amount of creotide. The pharmaceutically effective dose of the compounds or oligonucleotides of this disclosure will vary depending on the specific objective to be achieved, the age and physical condition of the patient being treated, the severity of the underlying disease, the duration of treatment, the nature of the concurrent treatments, and the specific compounds employed. For example, a therapeutically effective dose of the compounds or oligonucleotides of this disclosure administered to children or neonates may be atypically reduced according to reasonable medical judgment. Therefore, the effective dose of the compounds or oligonucleotides of this disclosure is the minimum amount that provides the desired effect.

[0370] The determined factual advantage of this disclosure is that the compound or oligonucleotide can be administered in a conventional manner, for example, by intravenous, intramuscular, subcutaneous, oral, or intraventricular injection routes, or by topical application in a cream or gel. Depending on the route of administration, the active ingredient, including the compound or oligonucleotide of this disclosure, may require coating within a material to protect the compound or oligonucleotide from the action of enzymes, acids, and other natural conditions that may inactivate it. For the purpose of administering the compound or oligonucleotide of this disclosure in a manner other than parenteral administration, the compound or oligonucleotide may be coated with a material or administered together with a material to prevent inactivation.

[0371] The compounds or oligonucleotides may be administered parenterally or intraperitoneally. Dispersions may also be prepared, for example, in glycerol, liquid polyethylene glycol, and mixtures thereof, as well as in oil.

[0372] Some examples of substances that can act as pharmaceutical carriers include sugars, e.g., lactose, glucose, and sucrose; starches, e.g., corn starch and potato starch; cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; tragacanth powder; malt; gelatin; talc; stearic acid; magnesium stearate; calcium sulfate; vegetable oils, e.g., peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa (theobroma) oil; polyols, e.g., propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; agar; alginic acid; pyrogen-free water; isotonic saline; and phosphate buffer solution; skim milk powder; and other non-toxic, suitable substances used in pharmaceutical formulations, e.g., vitamin C, estrogen, and echinacea. Wetting agents and lubricants, such as sodium lauryl sulfate, as well as colorants, flavoring and deodorizing agents, lubricants, additives, tableting agents, stabilizers, antioxidants, and preservatives may also be present. Solubilizers, such as creamaphores and β-cyclodextrin, may also be used in the pharmaceutical compositions herein.

[0373] Pharmaceutical compositions containing the active compounds or oligonucleotides (or their prodrugs) of this disclosure may be prepared by conventional mixing, dissolution, granulation, sugar coating, gelation, emulsification, encapsulation, capture, or lyophilization processes. These compositions may be formulated in a conventional manner using one or more physiologically acceptable carriers, diluents, additives, or auxiliaries that facilitate the processing of the active compounds into pharmaceutically usable preparations.

[0374] The pharmaceutical compositions of the subject matter of this disclosure may take a form suitable for virtually any method of administration, including, for example, topical, ocular, oral, buccal, systemic, nasal, injection, transdermal, rectal, or vaginal, or a form suitable for administration by inhalation or inhalation.

[0375] For topical administration, the active compound(s) or prodrug(s) may be formulated as a solution, gel, ointment, cream, or suspension.

[0376] Systemic formulations include formulations designed for administration by injection, such as subcutaneous, intravenous, intramuscular, intra-shelter, or intraperitoneal injection, and formulations designed for transdermal, transmucosal, oral, or pulmonary administration.

[0377] Useful injectable preparations include sterile suspensions, solutions, or emulsions of active compounds or oligonucleotides in aqueous or oily vehicles. The compositions may also contain formulation agents, such as suspending agents, stabilizers, and / or dispersants. Preparations for injection may be present in unit dosage forms (e.g., ampoules or multi-dose containers) and may contain added preservatives.

[0378] Alternatively, the injectable formulation may be provided in powder form for reconstitution before use with a suitable vehicle, including but not limited to sterile pyrogen-free water, buffer, and glucose solution. For this purpose, the active compound(s) or oligonucleotide(s) may be dried by techniques known in the art, such as lyophilization, and reconstituted before use.

[0379] For transmucosal administration, an appropriate permeabilizing agent is used in the formulation to address the barrier to which the drug must pass. Such permeabilizing agents are well known in the art.

[0380] For oral administration, the pharmaceutical composition may take the form of lozenges, tablets, or capsules, prepared by conventional means using pharmaceutically acceptable additives, such as binders (e.g., pregelatinized corn starch, polyvinylpyrrolidone, or hydroxypropyl methylcellulose); fillers (e.g., lactose, microcrystalline cellulose, or calcium hydrogen phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrants (e.g., potato starch or sodium starch glycolate); or wetting agents (e.g., sodium lauryl sulfate). These tablets may be coated, for example, with sugar or enteric coatings, by methods well known in the art.

[0381] Liquid preparations for oral administration may take the form of, for example, elixirs, solutions, syrups, or suspensions, or they may be provided as dry products to be prepared with water or other suitable vehicles before use. Such liquid preparations may be prepared by conventional means using pharmaceutically acceptable additives, such as suspending agents (e.g., sorbitol syrup, cellulose derivatives, or hydrogenated edible fats); emulsifiers (e.g., lectins or acacia); non-aqueous vehicles (e.g., almond oil, oily esters, ethyl alcohol, or fractionated edible oils); and preservatives (e.g., methyl p-hydroxybenzoate or propyl p-hydroxybenzoate, or sorbic acid). These preparations may also contain buffer salts, preservatives, flavoring agents, colorants, and sweeteners, as may be specified.

[0382] Preparations for oral administration can be appropriately formulated to provide a controlled release of the active compound or prodrug, as is well known.

[0383] For gus administration, the composition may take the form of tablets or lozenges, formulated in a conventional manner.

[0384] For rectal and vaginal administration routes, the active compound(s) or oligonucleotide(s) may be formulated as a liquid (for retained enemas), suppository, or ointment containing a conventional suppository base, such as cocoa butter or other glycerides.

[0385] For nasal administration, or administration by inhalation or inhalation, active compounds(s), oligonucleotides(s), or prodrugs(s) may be conveniently delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, fluorocarbon, carbon dioxide, or other suitable gas. In the case of pressurized aerosols, the dosage unit may be determined by providing a valve for delivering a measured amount. Capsules and cartridges for use in inhalers or ventilators (e.g., capsules and cartridges made of gelatin) may be formulated to contain a powder mixture of the compound or oligonucleotide with a suitable powder base such as lactose or starch.

[0386] Specific examples of aqueous suspension formulations suitable for nasal administration using commercially available nasal spray devices include the following components: active compound or prodrug (0.5-20 mg / ml); benzalkonium chloride (0.1-0.2 mg / ml); polysorbate 80 (TWEEN® 80; 0.5-5 mg / ml); carboxymethylcellulose sodium or microcrystalline cellulose (1-15 mg / ml); phenylethanol (1-4 mg / ml); and dextrose (20-50 mg / ml). The pH of the final suspension can be adjusted to a range of approximately pH 5 to pH 7, with approximately pH 5.5 being typical.

[0387] For ocular administration, active compounds, oligonucleotides, or prodrugs may be formulated as appropriate solutions, emulsions, and suspensions for ocular administration. Various vehicles suitable for administering compounds to the eye are known in the art. Specific, non-limiting examples are described in U.S. Patents 6,261,547; 6,197,934; 6,056,950; 5,800,807; 5,776,445; 5,698,219; 5,521,222; 5,403,841; 5,077,033; 4,882,150; and 4,738,851, each of which is incorporated herein by reference in whole.

[0388] For long-term delivery, active compounds(s), oligonucleotides(s), or prodrugs(s) may be formulated as depot preparations for administration by implantation or intramuscular injection. Active ingredients may be formulated using suitable polymer or hydrophobic materials (e.g., emulsions in acceptable oils) or ion-exchange resins, or as low-solubility derivatives, such as low-solubility salts. Alternatively, transdermal delivery systems may be used, manufactured as adhesive discs or patches that slowly release active compounds(s) or oligonucleotides(s) for transdermal absorption. For this purpose, penetration enhancers may be used to facilitate the transdermal penetration of active compounds(s) or oligonucleotides(s). Appropriate transdermal patches are described, for example, in U.S. Patent Nos. 5,407,713; 5,352,456; 5,332,213; 5,336,168; 5,290,561; 5,254,346; 5,164,189; 5,163,899; 5,088,977; 5,087,240; 5,008,110; and 4,921,475, each of which is incorporated herein by reference in whole.

[0389] Alternatively, other pharmaceutical delivery systems may be employed. Liposomes and emulsions are well-known examples of delivery vehicles that can be used to deliver active compounds, oligonucleotides, or prodrugs. Certain organic solvents, such as dimethyl sulfoxide (DMSO), may also be employed.

[0390] The pharmaceutical composition may be provided in a pack or dispenser device capable of containing one or more unit dosage forms, if desired, containing active compounds or oligonucleotides. The pack may, for example, be made of metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration.

[0391] The active compounds(s), oligonucleotides(s), or prodrugs(s) of this disclosure, or compositions thereof, are generally used in amounts effective to achieve the intended result, for example, in amounts effective to treat or prevent the specific disease being treated. The compounds(s) and oligonucleotides(s) may be administered therapeutically to achieve a therapeutic benefit or prophylactically to achieve a preventive benefit. A therapeutic benefit means the elimination or improvement of the underlying disorder being treated and / or the elimination or improvement of one or more symptoms associated with the underlying disorder, thereby causing the patient to report an improvement in sensation or condition, even though the patient may still have the underlying disorder. A therapeutic benefit also includes stopping or slowing the progression of the disease, whether or not improvement is achieved.

[0392] For prophylactic purposes, a compound or oligonucleotide may be administered to patients at risk of developing one of the previously described diseases. Patients at risk of developing a disease may be those who possess characteristics that place them in a designated group of patients at risk, as defined by an appropriate healthcare professional or group. Patients at risk may also be those who are typically or routinely in a background where the development of the underlying disease is likely to occur. In other words, patients at risk are those who are typically or routinely exposed to, or could actually be exposed to, the conditions that cause the disease or illness over a limited period of time. Alternatively, prophylactic administration may be given to patients diagnosed with an underlying disorder to avoid the development of symptoms.

[0393] The amount of compound administered depends on various factors, such as the specific indication being treated, the mode of administration, whether the desired benefit is preventive or therapeutic, the severity of the indication being treated, the patient's age and weight, and the bioavailability of the specific active compound. Determining the effective dosage is well within the capabilities of those skilled in the art.

[0394] The effective dosage can initially be estimated from in vitro assays. For example, the initial dosage for use in animals can be determined by the IC of a specific compound measured by in vitro assays, such as in vitro fungal MIC or MFC and other in vitro assays. 50 Or higher concentrations of the active compound in the circulating blood or serum may be achieved. Calculating the dosage to achieve such circulating blood or serum concentrations, taking into account the bioavailability of a particular compound, is well within the capabilities of those skilled in the art. For guidance, see "General Principles," Goodman and Gilman's *The Pharmaceutical Basis of Therapeutics*, Chapter 1, pp. 1-112, 13th ed., McGraw-Hill, and the references cited therein. These are incorporated herein by reference.

[0395] Initial dosages can also be estimated from in vivo data, such as animal models. Animal models useful for testing the efficacy of compounds to treat or prevent the various diseases described above are well known in the field.

[0396] The dosage typically ranges from approximately 0.0001 or 0.001 or 0.01 mg / kg / day to approximately 100 mg / kg / day, but among other factors, the compound itself is particularly important. The dose may be higher or lower, depending on the activity of the oligonucleotide, its bioavailability, the mode of administration, and the various factors discussed above. The dose and interval may be individually adjusted to provide sufficient plasma levels of the compound(s) or oligonucleotide(s) to maintain the therapeutic or prophylactic effect. In the case of topical administration or selective uptake, for example, local topical administration, the effective local concentration of the active compound(s) or oligonucleotide(s) may not be related to the plasma concentration. Those skilled in the art can optimize the effective local dose without excessive experimentation.

[0397] Compounds and oligonucleotides may be administered once daily, a few times or several times a day, or even multiple times a day, depending in particular on the indication being treated and the judgment of the prescribing physician.

[0398] Preferably, the compound(s) and oligonucleotide(s) provide a therapeutic or preventive benefit without causing substantial toxicity. The toxicity of the compound(s) and oligonucleotide(s) can be determined using standard pharmaceutical procedures. The dose-to-toxicity ratio is the therapeutic index. Compound(s) and oligonucleotide(s) exhibiting a high therapeutic index are preferred.

[0399] Any enumeration of chemical groups in any definition of a variable substance as described herein includes the definition of that variable substance as any single group or as a combination of the enumerated groups. Any descriptions of embodiments relating to a variable substance as described herein include that embodiment as any single embodiment or in combination with any other embodiment or part thereof. Any descriptions of embodiments as described herein include that embodiment as any single embodiment or in combination with any other embodiment or part thereof. [Examples]

[0400] For the purpose of enabling a more complete understanding of the invention as described herein, the following examples are provided. The examples described herein are given to illustrate the compounds, pharmaceutical compositions, and methods provided herein and should not be construed as limiting their scope in any way.

[0401] General experimental procedure The definition of a variable in a structure within a scheme as described herein is equivalent to that in the corresponding position in an equation illustrated herein.

[0402] Common abbreviations: ACN Acetonitrile br wide d double line DCM Dichloromethane dd double line double line DBA Dibenzylideneacetone DFAA (Difluoroacetic Acid Anhydrous) DIPEA Diisopropylethylamine DMF Dimethylformamide DMSO (Dimethyl Sulfoxide) dppf 1,1'-ferrocendiyl-bis(diphenylphosphine) HCl ethyl acetate h time HRMS high resolution mass spectrometry HPLC (High-Performance Liquid Chromatography) LCMS (Liquid Chromatography and Mass Spectrometry) MS mass spectrometry MW microwave m multiplet MeOH methanol min mL (milliliter) m / z mass-to-charge ratio NMP N-methyl-2-pyrrolidone NMR nuclear magnetic resonance ppm Parts Permillion rt or RT Room temperature s single line t triple line TFAA (Trifluoroacetic Anhydride) TLC (Thin-Layer Chromatography)

[0403] Example 1 Exemplary compounds falling within the scope of this disclosure could be synthesized according to the following scheme: [ka]

[0404] [ka]

[0405] 2-(3-phenoxypropyl)malonic acid (1) Dimethyl malonate (10 g, 75.68 mmol, 1.9 equivalents) was carefully added dropwise to a stirred solution of NaH (1.91 g, 47.8 mmol, 1.2 equivalents) in THF (50 mL) at room temperature. Upon addition, the reaction mixture became a concentrated suspension. THF (10 mL) was added to the reaction mixture and it was stirred for 1 hour. Benzyl 3-bromopropyl ether (9.126 g, 39.8 mmol, 1 equivalent) was added to the reaction mixture and the reaction mixture was heated to 85°C. After stirring for 5 hours, the reaction mixture was cooled to room temperature, diluted with SiO2 (100 mL), and washed with water (100 mL). The aqueous phase was extracted with SiO2 (100 mL). The combined organic layers were dried over Na2SO4. The solution was filtered and concentrated. The residue was purified by flash chromatography using 0-50% toluene in hexane to obtain 10.5 g (94%) as a clear oil. The product was confirmed by LC-MS and NMR. HPLC: rt = 4.996 min.

[0406] 2-(3-phenoxypropyl)malonic acid (2) LiOH was added to a stirred solution of compound 1 (10.5 g, 37.5 mmol, 1 equivalent) in THF / H2O (1:1, 100 mL). The reaction mixture was stirred overnight and then acidified to pH=2 with 1N HCl. The mixture was extracted with RINKAN (100 mL x 2), dried over Na2SO4, and concentrated to obtain 8.9 g of crude product 2 (93%) as a white solid, which was used in the next step without purification. HPLC: rt=3.592 min.

[0407] Example 2 Exemplary compounds falling within the scope of this disclosure could be synthesized according to the following scheme: [ka]

[0408] [ka]

[0409] 2-(3-((((benzyloxy)carbonyl)amino)propyl)dimethyl(2) malonate To a stirred solution of NaH (1.1 g, 27.6 mmol, 1.5 equivalents) in DMF (15 mL), dimethyl malonate (3.64 g, 27.6 mmol, 1.5 equivalents) in DMF (5 mL) was carefully added dropwise at room temperature. Upon addition, the reaction mixture became a concentrated suspension. DMF (5 mL) was added to this reaction mixture, and it was stirred at room temperature for 1 hour. Starting material 1 (5.0 g, 18.4 mmol, 1 equivalent) in DMF (5 mL) was added to this reaction mixture, and the reaction mixture was heated to 75°C. After stirring at 75°C for 5 hours, the reaction mixture was cooled to room temperature. The reaction mixture was diluted with SiO2 (100 mL) and washed with water (100 mL). The aqueous phase was extracted with SiO2 (100 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using 0-40% toluene in hexane to obtain 3.6 g of product 2 (60%) as a clear oil. The product was confirmed by LC-MS and NMR. HPLC: rt = 4.518.

[0410] 2-(3-(((benzyloxy)carbonyl)amino)propyl)malonatic acid (3) To a stirred solution of compound 2 (3.6 g, 11.13 mmol, 1 equivalent) in THF / H2O (1:1, 50 mL), LiOH (0.8 g, 33.4 mmol, 3 equivalents) was added. The reaction mixture was stirred overnight and then acidified to pH=2 with 1N HCl. The mixture was extracted with RINKAN (60 mL x 2), dried over Na2SO4, and concentrated to obtain 3.3 g of crude product 3 (100%) as a clear oil, which was used in the next step without purification. HPLC: rt=3.40 min.

[0411] Example 3 [ka] Alcohol 1 (5.0 g, 10.27 mmol, 1 equivalent), Na2CO3 (7.619 g, 71.888 mmol, 7 equivalents), and tetrabutylammonium bromide (0.132 g, 0.411 mmol, 0.04 equivalents) were dissolved in a two-phase mixture of CH2Cl2 (125 mL) and H2O (250 mL). Benzoyl chloride (1.550 mL, 13.351 mmol, 1.3 equivalents) was added to this reaction, and the reaction mixture was vigorously stirred overnight. The reaction mixture was extracted with DCM. The organic phase was dried over anhydrous Na2SO4, filtered, and evaporated under reduced pressure. The residue was dissolved in 1,2-dichloroethane (30 mL) and heated at 60°C for 20 minutes. This solution was stirred overnight at room temperature, the solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica gel using a gradient of 0–40% HCl in hexane to obtain 3.4 g (51%) of product (2) as a solid. LCMS:ESI+ m / z=591.1[M+H] + .

[0412] [ka] Dipropargylamine (2 g, 21.48 mmol, 1 equivalent) and K2CO3 (14.84 g, 107.4 mmol, 5 equivalents) were suspended in CH3CN (50 mL) and bromoacetyl chloride (1.788 mL, 21.48 mmol, 1 equivalent) was added at 0°C. The reaction mixture was stirred at 0°C for 1 hour, then at room temperature for 1 hour. The reaction mixture was diluted with DCM and washed with water. The organic phase was dried over Na2SO4, concentrated, and purified by column chromatography on silica gel using a gradient of 0-30% siRNA in hexane to obtain 2.35 g of product 3 (51%) as an oil. LCMS:ESI+ m / z=215.9[M+H] + .

[0413] [ka] To a stirred solution of alcohol 2 (2.4 g, 4.062 mmol, 1 equivalent) in DMF (8 mL), bromoacetamide 3 (1.304 g, 6.093 mmol, 1.5 equivalents) was added at 0°C, followed by the addition of NaH (0.211 g, 60% in mineral oil, 2.068 mmol, 1.3 equivalents). The reaction mixture was stirred at 0°C for 45 minutes. The mixture was then quenched by the addition of H2O. The reaction mixture was diluted with siRNA and washed with water. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using a gradient of 0-30% siRNA in hexane to obtain 2.17 g (74%) of product 4 as a white solid. LCMS:ESI+ m / z=725.2[M+H] + .

[0414] [ka] To a stirred solution of bis-alkyne 4 (2.91 g, 4.02 mmol, 1 equivalent) in THF (30 mL), Et3N·3HF (3.276 mL, 20.098 mmol, 5 equivalents) was added. The reaction mixture was stirred overnight at room temperature and then quenched by the addition of MeOH. The reaction mixture was diluted with RINKAN and washed with saturated aqueous NaHCO3, then brine. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using a gradient of 0–5% MeOH in RINKAN to obtain 1.85 g of alcohol 5 (95%) as a white solid. LCMS:ESI+ m / z=482.0[M+H] + .

[0415] [ka] To a stirred solution of alcohol 5 (1.85 g, 3.842 mmol, 1 equivalent) in pyridine (30 mL), DMTrCl (1.693 mg, 4.995 mmol, 1.3 equivalents) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched by the addition of MeOH, and the mixture was concentrated under reduced pressure. The resulting residue was partitioned between toluene and aqueous saturated NaHCO3. The organic layer was dried over Na2SO4, filtered, and The solution was concentrated. The resulting residue was purified by column chromatography on silica gel using a gradient of 0-50% siRNA in hexane to obtain 2.48 g of DMT ether 6 (82%) as a white solid. LCMS:ESI+ m / z=806.0[M+Na] + .

[0416] [ka] To a stirred solution of DMT ether 6 (780 mg, 0.995 mmol, 1 equivalent) in DMF (4 mL), NaH (52 mg, 60% in mineral oil, 1.294 mmol, 1.3 equivalents) was added at -10°C, followed by the addition of 2-bromo-N-(propa-2-in-1-yl)acetamide (263 mg, 1.493 mmol, 1.5 equivalents) in DMF (1 mL). The reaction mixture was stirred at -10°C for 20 minutes and then quenched by the addition of H₂O. The reaction mixture was partitioned between siRNA and water. The aqueous phase was extracted with siRNA, and the combined organic layers were dried over Na₂SO₄, filtered, and concentrated. The resulting residue was purified by column chromatography on silica gel using a gradient of siRNA from 0 to 60% siRNA in hexane to obtain 380 mg (43%) of product 7 as a white solid. LCMS:ESI+ m / z=901.7[M+Na] + .

[0417] [ka] To a solution of DMT ether 7 (0.38 g, 0.432 mmol, 1.00 equivalent) in DCM (5 mL), TFA (0.076 mL, 0.994 mmol, 2.30 equivalents) was added at room temperature, followed by the addition of Et3SiH (0.069 mL, 0.432 mmol, 1 equivalent). The reaction mixture was stirred at room temperature for 2 hours and then quenched with saturated aqueous NaHCO3. The product was extracted with DCM, and the organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography using a gradient of 0-10% MeOH in DCM to obtain 179 mg of alcohol 8 (71%). LCMS:ESI+ m / z=577.0[M+H] + .

[0418] [ka] To a solution of alcohol 8 (179 mg, 0.31 mmol, 1 equivalent) and azide 9 (0.562 g, 1.025 mmol, 3.3 equivalents) in THF (8 mL), a solution of CuSO4·5H2O (0.233 g, 0.931 mmol, 3 equivalents) in water (2 mL) was added at 0°C, followed by the addition of a solution of sodium ascorbate (0.215 g, 1.087 mmol, 3.5 equivalents) in water (2 mL). After stirring at 0°C for 5 minutes, the reaction mixture was warmed to room temperature and then stirred at room temperature for 45 minutes. The mixture was diluted with DCM and then washed with NaHCO3. The aqueous phase was extracted with DCM, and the organic layers were combined, dried over Na2SO4, filtered, and concentrated. The obtained residue was purified by column chromatography on silica gel using a gradient of 0-20% MeOH in DCM to obtain 398 mg of triazole 10 (58%) as a solid. MS: m / z = 2243.5 [M + Na] + .

[0419] [ka] A stirring solution of triazole 10 (100 mg, 0.045 mmol, 1 equivalent) and diisopropylethylamine (0.051 mL, 0.135 mmol, 6.5 equivalents) in anhydrous DCM (2 mL) was prepared under argon conditions by adding 2-cyanoethyl=N,N-diisopropylchlorophosphate. Sphoroamidite (0.030 mL, 0.293 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 30 minutes and then quenched with saturated NaHCO3 aqueous solution. The product was extracted by DCM, and the combined organic phase was washed with brine, dried over Na2SO4, filtered, and then concentrated. The resulting residue was purified by flash chromatography using a gradient of 0-5% MeOH in DCM in the presence of a base to obtain 71 mg of phosphoroamidite 11 (65%). MS: m / z = 2443.6 [M + H] + .

[0420] Example 4 [ka]

[0421] [ka] SEMCl (3.3 mL, 18.5 mmol, 1.2 equivalents) was added dropwise at 0°C to a solution of alcohol 1 (7.5 g, 15.4 mmol, 1 equivalent) and DBU (3.5 mL, 23 mmol, 1.5 equivalents) in DMF (50 mL). The reaction mixture was stirred overnight at room temperature, then diluted with ethyl acetate and hexane, and poured into cold water (500 mL). The layers were separated, and the organic layer was washed sequentially with water (2 × 300 mL) and then brine (200 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel (50 g) using a gradient of ethyl acetate in hexane from 0 to 25% to obtain compound 2 (7.5 g, 79%). LCMS:ESI+ m / z=639.4[M+Na] + .

[0422] [ka] Alcohol 2 (2.7 g, 4.38 mmol, 1 equivalent) was added to DMF (29 mL) and cooled to 0°C. t-butyl bromoacetate (1.9 mL, 13 mmol, 3 equivalents) was added, followed by sodium hydride (219 mg, 5.5 mmol, 1.25 equivalents). The reaction mixture was stirred at 0°C for 1 hour, and the remaining base was quenched at 0°C by adding methanol (1.5 mL). The product was extracted with ethyl acetate, and the organic layer was washed with water and then brine. The organic phase was dried over sodium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel (50 g) using a gradient of ethyl acetate in hexane from 0 to 20% to obtain ester 3 (3.13 g, 98%) as an oil. LCMS:ESI+ m / z=753.4[M+Na] + .

[0423] [ka] TBAF (1.0 M, 12.7 mL, 3.9 mmol, 3 equivalents in THF) was added dropwise at 0°C to a solution of disiloxane 3 (3.1 g, 4.24 mmol, 1 equivalent) in THF (42 mL, 0.1 M). After 2 hours at 0°C, the reaction mixture was poured into a saturated solution of sodium bicarbonate (200 mL) and extracted with DCM (2 × 150 mL). The combined organic extracts were washed with water (200 mL) and then brine (150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel (25 g) using a gradient of ethyl acetate in hexane from 0 to 70% to obtain diol 4 (1.82 g, 88%) as an oil. LCMS:ESI+ m / z=511.0[M+Na] + . [ka]

[0424] TBSCl (666 mg, 4.4 mmol, 1.2 equivalents) as a solid, diol (4 ) (1.8 g, 3.7 mmol, 1 equivalent) and imidazole (501 mg, 7.4 mmol, 2 equivalents) were added in one batch at 0°C to a solution in DMF (18 mL). The reaction mixture was stirred at room temperature for 2.5 hours, diluted with ethyl acetate, and then poured into stirred cold water (200 mL). The organic layer was washed with water (2 × 200 mL) and then with brine (150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel (50 g) using 0-25% ethyl acetate in hexane to obtain alcohol 5 (1.94 g, 87%) as an oil. LCMS:ESI+ m / z=625.2[M+Na] + .

[0425] [ka] Alcohol 5 (1.94 g, 3.14 mmol, 1 equivalent) was added to anhydrous DMF (21 mL) and cooled to 0°C. Propargyl bromide (80% w / w in toluene, 0.7 mL, 6.29 mmol, 2 equivalents) was added, followed by the addition of NaH (157 mg, 3.93 mmol, 1.25 equivalents) as a solid in one batch. After 90 minutes at 0°C, a further amount of propargyl bromide (80% w / w in toluene, 0.35 mL, 3.1 mmol, 1 equivalent) was added, followed by the addition of NaH (63 mg, 1.57 mmol, 0.5 equivalents). After another hour, the reaction mixture was diluted with ethyl acetate and washed with water (2 × 200 mL) and then brine (150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel (25 g) using a gradient of ethyl acetate in hexane (0-15%) to obtain alkyne (6) (1.38 g, yield 68%) as an oil. LCMS:ESI+ m / z=663.3[M+Na] + .

[0426] [ka] TFA (2.5 mL) was added dropwise at 0°C to a solution of t-butyl ester (6) (0.98 g, 1.53 mmol, 1 equivalent) in THF (10 mL) and H2O (2.5 mL). The reaction mixture was stirred at room temperature for 1.5 hours and then concentrated. The resulting residue was dissolved in dichloromethane (10 mL), cooled to 0°C, and then TFA (5 mL) was added. After 1.5 hours, the reaction mixture was concentrated under reduced pressure, and the resulting residue was diluted with DCM (2 mL) and toluene (2 mL) and then concentrated under reduced pressure. The resulting residue was dissolved in DMF (5 mL), cooled to 0°C, and then DIPEA (1.3 mL, 7.65 mmol, 5 equivalents) and dipropargylamine (0.8 mL, 7.65 mmol, 5 equivalents) were added. Equivalents of α7 and HATU (1.098 g, 2.89 mmol, 1.9 equivalents) were added sequentially. The reaction mixture was stirred at room temperature for 75 minutes, and then diluted with ethyl acetate and 1N HCl (50 mL). The mixture was stirred vigorously for 5 minutes, and then the organic phase was separated. The aqueous layer was further extracted with ethyl acetate. The combined organic extract was washed with water (2 × 50 mL), then with brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash column chromatography on silica gel (25 g) using a gradient of ethyl acetate in hexane from 0 to 95% to obtain amide 7 (532 mg, 84%) as a viscous solid. LCMS:ESI+ m / z=416.1[M+H] + .

[0427] [ka] A solution of sodium ascorbate (90 mg, 0.455 mmol, 3.5 equivalents) in water (1.5 mL) and a solution of copper sulfate pentahydrate (97 mg, 0.39 mmol, 3 equivalents) in water (1.5 mL) were sequentially added dropwise to a solution of alkyne 7 and azide 8 in THF at 0°C. After 5 minutes, the reaction mixture was stirred at room temperature for 3 hours, then diluted with DCM (50 mL) and washed with aqueous saturated sodium bicarbonate (50 mL). The aqueous layer was extracted with DCM (3 × 25 mL), the combined organic extracts were dried over anhydrous sodium sulfate and then filtered through Celite. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by flash column chromatography on silica gel (5 g) using a methanol gradient of 0–75% in ethyl acetate to obtain tris-triazole 9 (120 mg, 45%) as a sticky solid. LCMS:ESI+ m / z=1031.0[(M+2H) / 2] + .

[0428] [ka] Compound 10 was synthesized from Compound 9 following the same procedure as in Example 3. LCMS:ESI+ m / z=1089.4[(M-NH(iPr)2+H2O+2H) / 2] + .

[0429] Example 5 Synthesis of the asymmetric tri-antennary 3'-triazole GalNAc-linker: [ka]

[0430] [ka]

[0431] Synthesis of asymmetric three-branched 5'-GalNAc-triazole linker: [ka]

[0432] [ka]

[0433] Synthesis of asymmetric three-branched 3'-GalNAc-triazole and amide linker: [ka]

[0434] Exemplary compounds falling within the scope of this disclosure could be synthesized according to the following scheme: [ka]

[0435] Synthesis of the asymmetric three-branched 3'-triazole GalNAc-linker: [ka]

[0436] [ka]

[0437] Synthesis of asymmetric three-branched 3'-GalNAc-amide linker: [ka]

[0438] [ka]

[0439] [ka]

[0440] Exemplary compounds falling within the scope of this disclosure could be synthesized according to the following scheme: [ka]

[0441] Example 6 [ka]

[0442] Intermediate 2 (INT-2). [ka] To a solution of INT-1 (8.5 g, 12.94 mmol, 1 equivalent) in siRNA (200 mL) and TFA (3 mL), Pd / C (1.377 g, 10 wt%, 1.294 mmol, 0.1 equivalent) was added under argon. This round-bottom flask was flushed with H2 gas, and the suspension was stirred overnight under H2 (1 atm). The catalyst was filtered off through Celite, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography using a gradient of 0-20% MeOH in DCM to obtain INT-2 (5.9 g, 74%).

[0443] Intermediate 3 (INT-3). [ka] To a solution of INT-2 (3.5 g, 5.649 mmol, 1 equivalent) and 3-azidopropionic acid (0.975 g, 8.474 mmol, 1.5 equivalents) in DCM (50 mL), HATU (3.222 g, 8.474 mmol, 1.5 equivalents) and then DIPEA (9.84 mL, 56.492 mmol, 10 equivalents) were added dropwise at 0°C. The reaction mixture was stirred at 0°C for 5 minutes, then at room temperature for 1 hour. The reaction was quenched by the addition of water, and the product was partitioned between DCM and H2O. The aqueous phase was extracted with DCM, and the combined organic layers were dried over Na2SO4, filtered, and concentrated. The resulting residue was purified by silica gel column chromatography using a gradient of 0-10% MeOH in siRNA to obtain INT-3 (3.27 g, 93%).

[0444] Intermediate 5 (INT-5). [ka] To a stirred solution of INT-4 (1 g, 1.832 mmol, 1 equivalent) in anhydrous DMF (15 ml) at 0°C, 60 w% NaH (220 mg, 5.495 mmol, 3 equivalents) was added. This reaction mixture was stirred at 0°C for 30 minutes, then propargyl bromide (0.68 mL, 4.579 mmol, 2.5 equivalents) was added dropwise, and the mixture was stirred at room temperature for 12 hours. This reaction mixture was quenched with water (20 mL), extracted with ethyl acetate (3 × 50 mL), dried to (Na₂SO₄), and then concentrated. The resulting residue was purified by silica gel column chromatography using a gradient of ethyl acetate in hexane from 0 to 50% as the eluate to obtain INT-5 (919 mg, 80%) as a solid. LCMS ESI+: m / z = 683[M+Na]+.

[0445] Intermediate 6 (INT-6). [ka] INT-5 (400 mg, 0.606 mmol, 1.00 equivalent) in a 10 mL dose (DCM) To the solution, TFA (0.1 mL, 1.33 mmol, 2.20 equivalents) and Et3SiH (0.1 mL, 0.667 mmol, 1.1 equivalents) were added sequentially at room temperature. The reaction mixture was stirred at room temperature for 5 hours and then diluted with DCM (50 mL). The organic phase was washed with NaHCO3 (50 mL) and then brine (50 mL), dried to (Na2SO4), filtered, and concentrated. The resulting residue was purified by silica gel column chromatography using a gradient of 0–100% siRNA in hexane to obtain INT-6 (200 mg, 50%) as a solid. LCMS ESI+: m / z = 359[M+H]+.

[0446] Intermediate 7 (INT-7). [ka] To a stirred mixture of INT-6 (8 mg, 0.022 mmol, 1 equivalent) and INT-3 (62 mg, 0.101 mmol, 4.5) in THF (1 mL), a solution of CuSO4·5H2O (17 mg, 0.067 mmol, 3 equivalents) in water (0.25 mL) was added at 0°C, followed by the addition of a solution of sodium ascorbate (15 mg, 0.078 mmol, 3.5 equivalents) in water (0.25 mL). This mixture was stirred at room temperature for 1 hour, diluted with saturated aqueous NaHCO3 (10 mL), and then extracted with DCM (2 × 20 mL). The organic phases were combined, washed with brine (20 mL), dried, filtered, and concentrated. The resulting crude residue was purified by silica gel column chromatography using a gradient of 0–30% MeOH in DCM to obtain INT-7 (20 mg 40%) as a solid. LCMS ESI+: m / z = 2239[M+Na]+.

[0447] Example 6. [ka] To a solution of INT-7 (70 mg, 0.032 mmol, 1 equivalent) in DCM (10 ml), add diisopropylethylamine (0.013 mL, 0.076 mmol, 2.4 equivalents) at 0°C, then add cyanoethyl=N,N-diisopropylchlorophosphoramidite ( 0.008 mL, 0.038 mmol, 1.2 equivalents) were added dropwise in sequence. The reaction mixture was stirred at room temperature for 2 hours, followed by further diisopropylethylamine (0.013 mL, 0.076 mmol, 2.4 equivalents), and then cyanoethyl=N,N-di Sopropyl chlorophosphoramidite (0.008 mL, 0.038 mmol, 1.2 equivalents) was added dropwise. The mixture was stirred at room temperature for 24 hours, diluted with saturated NaHCO3 aqueous solution (20 mL), and the product was extracted with DCM (2 × 50 mL). The organic phase was dried (Na2SO4), filtered, and concentrated. The resulting residue was purified by silica gel column chromatography using a gradient of 0-30% MeOH in DCM to obtain Example 6 (10 mg, 26%) as a solid. LCMS ESI+: m / z = 2439[M+Na]+. 1 H and 31 The 1P NMR spectrum matches the structure of the desired product.

[0448] Example 7 [ka]

[0449] [ka]

[0450] [ka] To a stirred, cooled (0°C) solution of Compound 1 (10.08 g, 18.462 mmol, 1 equivalent) in anhydrous DMF (100 ml), NaH 60 w% (2.58 mg, 64.615 mmol, 3 equivalents) was added. The reaction mixture was stirred at 0°C for 30 minutes. Propargyl bromide (8.236 mL, 55.38 mmol, 3 equivalents) was added dropwise, and stirring was continued at 0°C for 2 hours. The reaction mixture was quenched with water (200 mL), extracted with ethyl acetate (200 mL), washed with 2 × 100 mL of water and 100 mL of brine, dried (Na2SO4), concentrated, and the residue was purified by silica gel column chromatography using 0–70% ethyl acetate / hexane as the eluent. The pure fractions were combined and concentrated to obtain tri-propargyl 2 (12.1 g, 99%) as a white solid. The product was identified by NMR and LC-MS (m / z 684 M+Na).

[0451] [ka] DMT ether 2 (4.1g, 6.212 mmol, 1.00 equivalent) in DCM (30m To the solution in (L), TFA (1.09 mL, 14.288 mmol, 2.30 equivalents) was added. The color of the solution changed to red. Et3SiH (1.09 mL, 6.833 mmol, 1.1 equivalents) was added at room temperature. This reaction mixture was stirred at room temperature for 5 hours. LC-MS showed complete deprotection. This reaction mixture was diluted with aq. saturated NaHCO3 (100 ml), extracted with DCM (2 × 200 ml), and the organic phase was washed with NaHCO3 (50 ml) and brine (100 ml), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography using a gradient of 0-100% siRNA in hexane as the eluent to obtain the desired product 3 as a solid (1.35 g, 61%). LC-MS ESI+ m / z 359[M+H] + .

[0452] [ka] A mixture of commercially available amino alcohol 6 (50.0 g, 258.75 mmol, 1 equivalent) in 250 mL of toluene (250 mL) and 250 mL of H2O was prepared by adding NaHCO3 (30.4 g, 362.25 mmol, 1.4 equivalents), followed by the addition of CbzCl (36.8 mL, 258.75 mmol, 1 equivalent) at room temperature. The mixture was stirred overnight at room temperature. Workup: The organic layer was separated, the aqueous phase was extracted with toluene (80 mL x 2), and then dried over MgSO4. The residue was purified by flash chromatography using 30%-50% toluene / hept to obtain compound 7 (72 g, 85% yield) as a syrup.

[0453] [ka] To a suspension of commercially available GalNAc acetate (10 g, 25.68 mmol, 1 equivalent) in 1,2-dichloroethane (50 mL), TMSOTf (6.5 mL, 35.96 mmol, 1.4 equivalents) was added over 3 minutes at room temperature. The mixture was then heated to 50°C and stirred at 50°C for 5 hours. Compound 7 (11.8 g, 35.957 mmol, 1.4 equivalents) and molecular sieves of 4A (10 g) were added, followed by the addition of TMSOTf (1.2 ml, 6.42 mmol, 0.25 equivalents). The mixture was stirred at room temperature for 2 days. The mixture was filtered, the filtrate was washed with brine (100 mL) and Sat NaHCO3 (50 mL), concentrated, and the residue was purified by flash column chromatography.

[0454] [ka] To a solution of GalNAc-PEG4-NHCbz 8 (8.5 g, 12.9 mmol, 1 equivalent) in SiO2 (200 ml) and TFA (3 ml), Pd / C (1.38 g, 10 wt%, 1.3 mmol, 0.1 equivalent) was added. This suspension was stirred overnight under H2 (1 atm). The reaction mixture was then filtered through Celite and concentrated. The residue was purified by silica gel column chromatography using a gradient of 0-20% MeOH in DCM to obtain GalNAc-PEG4-NH2.TFA 9 (5.90 g, 7 4% was obtained as an adhesive solid.

[0455] [ka] To a solution of GalNAc-PEG4-NH2.TFA 9 (6.0 g, 9.6 mmol, 1 equivalent) and 3-azidoacetic acid (1.6 g, 14.5 mmol, 1.5 equivalents) in DCM (100 ml), HATU (5.5 g, 14.5 mmol, 1.5 equivalents) was added at 0°C, followed by the addition of DIEA (16.9 ml, 96.8 mmol, 10 equivalents). The reaction mixture was stirred at 0°C for 5 minutes, then warmed to room temperature and stirred for 4 hours. The reaction was quenched by the addition of H2O. The crude reaction mixture was partitioned between DCM and 1N HCl. The aqueous phase was extracted with DCM. The combined organic layers were dried over Na2SO4, filtered, and then concentrated. The residue was purified by silica gel column chromatography using a gradient of 0-10% MeOH in HCl to obtain the desired product 10 (4.95 g, 83%) as an oil. LCMS ESI+ m / z=620[M+H] + .

[0456] [ka] To a stirred, cooled 0°C solution of alkyne 3 (585 mg, 1.634 mmol, 1 equivalent) and azide 10 (3.136 g, 5.066 mmol, 3.1 equivalents) in THF (40 mL), CuSO4·5H2O (0.408 g, 1.634 mmol, 1 equivalent) in water (10 mL) was added, followed by the addition of sodium ascorbate (0.485 g, 2.451 mmol, 1.5 equivalents) in water (10 mL). The mixture was stirred at room temperature for 1 hour. LC-MS showed product formation and complete disappearance of the alkyne. The reaction mixture was diluted with aq. saturated NaHCO3 (100 mL), extracted with DCM 2 × 200 mL, and washed with brine solution (100 mL). The combined extracts were dried over Na2SO4 and concentrated. The resulting crude product was purified by column chromatography using 0-30% MeOH / DCM as the eluate. The pure fractions were combined and concentrated. The resulting solid was co-evaporated with toluene and dried under high vacuum to obtain tris-triazole 4 (2.75 g 75%) as a yellow solid. LC-MS (m / z 1109 M+ / 2) and NMR correspond to the product.

[0457] [ka] Alcohol 4 (2.7g, 1.218 mmol, 1 equivalent) and diisopropyl ethyl To a stirred solution of amine (1.37 mL, 7.9 mmol, 6.5 equivalents) in DCM (20 mL), N,N-diisopropyl chlorophosphoramidite (0.8 mL, 3.65 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 15 minutes. LC-MS and HPLC indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (100 mL), extracted with DCM (2 × 100 mL), washed with brine (100 mL), dried over N2SO4, and the crude product was evaporated and loaded onto a pre-equalized (2% Et3N-DCM) Biotage silica gel column (50 g, 20 μm). The crude product was purified by flash chromatography using 0-10% MeOH / DCM containing 2% Et3N as an additive. The pure fractions were combined, concentrated, and dried under high vacuum to obtain phosphoramidite 5 (2.75 g 93%) as a white solid. Purity was 91% by HPLC, P31-NMR, mass (m / z 2439 M+Na), H1-NMR, and LC-MS, and HPLC correspond to the product.

[0458] Example 8 [ka]

[0459] [ka]

[0460] [ka]

[0461] [ka]

[0462] Alcohol 1 (10.0 g, 20.54 mmol, 1 equivalent), Na2CO3 (15.239 g, 143.776 mmol, 7 equivalents), and tetrabutylammonium bromide (0.265 g, 0.822 mmol, 0.04 equivalents) were stirred in a two-phase mixture of CH2Cl2 (200 mL) and H2O (400 mL). Benzoyl chloride (3.099 mL, 26.71 mmol, 1.3 equivalents) was added to the reaction mixture, and the reaction mixture was vigorously stirred for 6 hours. The reaction mixture was diluted with CH2Cl2. The aqueous phase was separated and extracted with DCM. The combined organic layers were dried over anhydrous Na2SO4, filtered, and evaporated under reduced pressure. The residue was dissolved in 1,2-dichloroethane (50 mL) and heated at 60°C for 20 minutes. The solution was then stirred overnight at room temperature. The solvent was removed under reduced pressure, and the residue was purified by column chromatography using silica gel with 0-30% RINKAN in hexane to obtain 7.3 g (60%) of product 2 as a white solid. Compound 3 is the main byproduct.

[0463] [ka] To a suspension of dipropargylamine (5 g, 53.688 mmol, 1 equivalent) and K2CO3 (37.1 g, 268.44 mmol, 5 equivalents) in CH3CN (100 mL), bromoacetyl chloride (4.471 mL, 53.688 mmol, 1 equivalent) was added at 0°C. The reaction mixture was stirred at 0°C for 1 hour. LC-MS indicated that the reaction was complete. The reaction mixture was diluted with DCM and washed with water. The organic phase was dried over Na2SO4, concentrated, and purified by CC (0% → 30% toluene-hexane). The mixture was prepared to obtain 6.7 g of product 4 (58%) as a yellow oily substance.

[0464] [ka] To a stirred solution of alcohol 2 (7.1 g, 12.017 mmol, 1 equivalent) in DMF (25 mL) at 0°C, amide 4 (3.86 g, 18.026 mmol, 1.5 equivalents) was added, followed by the addition of NaH (0.625 g, 60% of mineral oil, 15.622 mmol, 1.3 equivalents) in two portions. The reaction mixture was stirred at 0°C for 45 minutes, and then quenched by the addition of H2O. The reaction mixture was diluted with siRNA and washed with water. The aqueous phase was extracted with siRNA. The combined organic layers were dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using 0-40% siRNA in hexane to obtain 7.1 g (81%) of product 5 as a white solid.

[0465] [ka] To a stirred solution of bis-alkyne 5 (7.2 g, 9.945 mmol, 1 equivalent) in THF (100 mL), Et3N·3HF (6.484 mL, 39.781 mmol, 4 equivalents) was added. The reaction mixture was stirred overnight at room temperature and then quenched by the addition of MeOH. The reaction mixture was diluted with RINKAN and washed with NaHCO3 and brine. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using 0-5% MeOH in RINKAN to obtain 4.5 g of product 6 (94%) as a white solid.

[0466] [ka] Alcohol 6 was dried by azeotropic distillation with toluene (1 × 50 mL). To a stirred solution of this alcohol 6 (6.4 g, 13.293 mmol, 1 equivalent) in anhydrous pyridine (50 mL), DMTrCl (5.855 g, 17.281 mmol, 1.3 equivalents) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched by adding MeOH. The crude reaction mixture was concentrated under reduced pressure. The residue was partitioned between HCl and NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified twice with 0-50% HCl in hexane to obtain 9.8 g of product 7 (88%) as a white solid.

[0467] [ka] To a stirred solution of DMTr ether 7 (6.0 g, 7.655 mmol, 1 equivalent) in DMF (40 mL), NaH (459 mg, 60% in mineral oil, 11.482 mmol, 1.5 equivalents) was added at -10°C, followed immediately by the addition of 2-bromo-N-(propa-2-in-1-yl)acetamide (2.695 g, 15.309 mmol, 2 equivalents; Synthonix) in DMF (5 mL). The reaction mixture was stirred at -10°C for 30 minutes. The reaction was then quenched by the addition of H2O. The reaction mixture was diluted with ethyl acetate and washed with water. The aqueous phase was extracted with ethyl acetate (twice). The combined organic layers were dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using 0-60% ethyl acetate in hexane to obtain 3.56 g (53%) of product 8 as a white solid.

[0468] [ka] To a solution of DMT ether 8 (3.56 g, 4.05 mmol, 1.00 equivalent) in DCM (50 mL), TFA (0.93 mL, 12.15 mmol, 3.0 equivalent) was added at room temperature. The reaction mixture was stirred at room temperature for 90 minutes. Further TFA (0.31 mL, 4.05 mmol, 1.0 equivalent) was added to the reaction mixture, and the mixture was stirred at room temperature for 30 minutes. The reaction was quenched by the careful addition of aq. NaHCO3. The reaction mixture was partitioned between saturated NaHCO3 and DCM. The aqueous phase was extracted with DCM (once). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0-10% MeOH in DCM to obtain 1.89 g of product 9 (81%) as a white solid.

[0469] [ka] A suspension of commercially available GalNAc acetate (64 g, 164.20 mmol, 1.2 equivalents) in 1,2-dichloroethane (300 mL) was to which TMSOTf (29.7 mL, 164.21 mmol, 1.2 equivalents, d: 1.228) was added over 5 minutes at room temperature. The mixture was then heated to 50°C and stirred at 50°C for 5 hours. The mixture was cooled to room temperature, then 4A MS (60 g) was added, followed by the addition of commercially available HO-PEG-6-N3 (30.0 g, 136.84 mmol, 1.0 equivalent). The mixture was stirred at room temperature for 10 minutes. Later, TMSOTf (7.4 mL, 41.05 mmol, 0.3 equivalents) was added. The resulting mixture was stirred at room temperature for 2 days. Workup: The reaction mixture was filtered and washed with DCM (50 mL). The filtrate was poured into brine (100 mL) + Sat NaHCO3 (150 mL). The organic layer was separated, and the aqueous phase was extracted with DCM (50 mL). The combined organic layers were washed with brine (100 mL) + Sat NaHCO3 (100 mL) and concentrated. The residue was purified by flash chromatography using 50% alkyl / Hept, then alkyl, then 1%-6% MeOH / alkyl. 37.2 g (60%) of azide 12 was obtained as a pale yellow syrup.

[0470] [ka] To a solution of alcohol 9 (2.0 g, 3.469 mmol, 1 equivalent) and azide 12 (6.089 g, 11.1 mmol, 3.2 equivalents) in THF (30 mL), a solution of CuSO4·5H2O (0.260 g, 1.041 mmol, 0.3 equivalents) in water (7.5 mL) and a solution of sodium ascorbate (0.309 g, 1.561 mmol, 0.45 equivalents) in water (7.5 mL) were added at 0°C. After stirring at 0°C for 5 minutes, the reaction mixture was warmed to room temperature. The reaction mixture was stirred at room temperature for 3 hours. LC-MS indicated that the reaction was complete. The reaction mixture was diluted with DCM and washed with NaHCO3. The aqueous phase was extracted with DCM (4 times). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified with 0-20% MeOH in DCM to obtain 6.46 g of triazole 10 (84%) as a yellow solid.

[0471] [ka] A stirred solution of triazole 10 (6.1 g, 2.745 mmol, 1 equivalent) and diisopropylethylamine (3.108 mL, 17.843 mmol, 6.5 equivalents) in anhydrous DCM (100 mL) is prepared by adding 2-cyanoethyl=N,N-diisopropylchlorophosphorole Midite (1.837 mL, 8.235 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 45 minutes. LC-MS and HPLC indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 solution and partitioned between DCM and saturated NaHCO3. The DCM phase was collected. The aqueous phase was extracted with DCM (once). The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was collected in pre-equilibrated (2% Et3N-DCM) biotage silica gel. The sample was loaded onto a ram (50 g, 20 μm) and purified by flash chromatography using 0-5-10% MeOH / DCM containing 2% Et3N as an additive. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 5.51 g of phosphoramidite 11 (83%) (HPLC purity 97%).

[0472] Example 9 [ka]

[0473] [ka] SEMCl (3.3 mL, 18.5 mmol, 1.2 equivalents) was added dropwise at 0°C to a solution of alcohol 1 (7.5 g, 15.4 mmol, 1 equivalent) and DBU (3.5 mL, 23 mmol, 1.5 equivalents) in DMF (50 mL). The reaction mixture was placed in an ice bath and slowly warmed to room temperature overnight. After 16 hours, the reaction mixture was diluted with ethyl acetate:hexane (5:1, 350 mL) and poured into cold stirred water (500 mL). The organic layer was washed with water (2 x 300 mL) and saturated sodium chloride aqueous solution (200 mL). The aqueous layer was extracted once separately with ethyl acetate-hexane (5:1, 250 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting colorless oily substance (10 g) was purified by flash column chromatography using silica gel (50 g, gradient: ethyl acetate in hexane, 0 → 15% [3 CV], 15% [3 CV], 15 → 20% [4 CV], 20% [2 CV], 20 → 25% [3 CV], 25% [4 CV]) to obtain alcohol 2 as a colorless oily substance (7.5 g, 79%). TLC (25% ethyl acetate in hexane). LCMS: ESI+ m / z 639.45 [M+Na] + .

[0474] [ka] Alcohol 2 (7.5 g, 12.2 mmol, 1 equivalent) was dried azeotropically by concentration at 35°C under reduced pressure from anhydrous toluene (20 mL). The resulting oily substance was dissolved in DMF (50 mL) and cooled to 0°C. 4.5 mL, 26 mol, 2.5 equivalents of t-butyl bromoacetate were added, followed by sodium hydride (60% dispersion in mineral oil, 608 mg, 15.2 mmol, 1.25 equivalents). This reaction mixture was maintained at 0°C. After 70 minutes, a further amount of sodium hydride (230 mg, 5.8 mmol, 0.5 equivalents) was added as a solid all at once. After a total reaction time of 2.5 hours, this reaction mixture was quenched with methanol (2 mL) and immediately poured into a stirring mixture of ethyl acetate-hexane (4:1, 250 mL) and a 25% saturated sodium chloride aqueous solution (500 mL). The organic layer was washed with 10% saturated sodium chloride aqueous solution (2 × 250 mL), water (100 mL), and saturated sodium chloride aqueous solution (100 mL). The aqueous layer was extracted once with ethyl acetate-hexane (4:1, 250 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting colorless residue (12.2 g) was purified by flash column chromatography on silica gel (eluate: ethyl acetate in hexane, gradient: 0% [2 CV], 0 → 5% [2 CV], 5% [1 CV], 5 → 10% [5 CV], 10% [1 CV], 10 → 15% [3 CV, less elutes first, then the major component], 15% [1 CV], 15 → 20% [1 CV], 20% [5 CV]) to obtain ester 3 (7.8 g, 88%) as a colorless oil. TLC (20% toluene / hexane). LCMS:ESI+ m / z 753.41[M+Na] + .

[0475] [ka] TBAF (1.0 M, 30 mmol, 2.8 equivalents in THF) was added dropwise at 0°C to a solution of disiloxane 3 (7.8 g, 10.7 mmol, 1 equivalent) in THF (100 mL, 0.1 M). After 6 hours at 0°C, the reaction mixture was poured into a saturated aqueous solution of sodium bicarbonate (250 mL) and extracted with DCM (3 × 150 mL). The combined organic extracts were washed with water (250 mL) and saturated aqueous sodium chloride solution (150 mL). The aqueous layer was separately extracted once with DCM (150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting colorless oily substance (10 g) was purified by flash column chromatography using silica gel (50 g, gradient: ethyl acetate in hexane, 0 → 60% [7 CV], 60 → 75% [2 CV], 75% [2 CV], 75 → 85% [2 CV], 85% [4 CV]) to obtain diol 4 (4.2 g, 81%) as a sticky white foam. LCMS: ESI+ m / z 511.09 [M + Na] + .

[0476] [ka] Diol 4 (4.2 g, 8.6 mmol, 1 equivalent) was dried azeotropically by concentration at 35°C from anhydrous DCM (10 mL) and toluene (20 mL). TBSCl (1.56 g, 10.4 mmol, 1.2 equivalents) was added as a solid to the solution of diol 4 and imidazole (1.17 g, 17.2 mmol, 2 equivalents) in DMF (40 mL) at 0°C. The reaction mixture was removed from the ice bath and stirred at room temperature. After 20 hours, the reaction mixture was diluted with ethyl acetate-hexane (5:1, 250 mL) and then poured into stirred cold water (400 mL). The organic layer was washed with water (3 × 300 mL) and saturated sodium chloride aqueous solution (250 mL). The aqueous layer was extracted once separately with ethyl acetate-hexane (5:1, 150 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained crude residue (6.6 g) was purified by flash column chromatography on silica gel (50 g, 60 μM, gradient: ethyl acetate in hexane, 0 → 10% [3 CV], 10% [6 CV], 10 → 25% [5 CV], 25% [4 CV]) to obtain alcohol 5. The mixed fraction (1.1 g) was purified by flash column chromatography on silica gel (25 g, 60 μM, gradient: ethyl acetate in hexane, 0 → 5% [3 CV], 5% [5 CV], 5 → 10% [8 CV], 10% [5 CV]) to obtain further alcohol 5. This was combined with the previously pure fraction to obtain alcohol 5 (4.43 g, yield 85.5%) as a colorless, viscous oil. TLC (25% siRNA / hexane). LCMS: ESI+ m / z 625.25 [M+Na] + .

[0477] [ka] Alcohol 5 (4.43 g, 7.18 mmol, 1 equivalent) was dried by azeotropy after concentration under reduced pressure from anhydrous toluene (10 mL). The resulting residue was dissolved in anhydrous DMF (35 mL, 0.2 M) and cooled to 0°C. Propargyl bromide (80% in toluene, 2.4 mL, 22 mmol, 3 equivalents) was added, followed by the single addition of NaH (431 mg, 10.8 mmol, 1.5 equivalents) as a solid. After 1 hour, further propargyl bromide (80% w / w in toluene, 1.6 mL, 11 mmol, 1.5 equivalents) was added, followed by the single addition of NaH (144 mg, 3.6 mmol, 0.5 equivalents) as a solid. After 1 hour (total reaction time 2 hours), the reaction mixture was diluted with ethyl acetate-hexane (5:1, 350 mL) and washed with 10% saturated brine (3 × 350 mL) and saturated sodium chloride aqueous solution (250 mL). The aqueous layer was extracted once separately with ethyl acetate-hexane (5:1, 150 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting brown oily substance (5.2 g) was purified by flash column chromatography on silica gel (50 g, 60 μm, eluate: ethyl acetate in hexane, 0% [2 CV], 0 → 7% [5 CV], 7% [4 CV], 7 → 12% [3 CV], 12% [2 CV], 12 → 15% [2 CV], 15% [3 CV]) to obtain alkyne 6 (3.85 g, 83.7%) as a pale yellow oily substance. LCMS:ESI+ m / z 663.36[M+Na] + .

[0478] [ka] TFA (2.5 mL) was added dropwise at 0°C to a solution of alkyne 6 (0.98 g, 1.53 mmol, 1 equivalent) in THF (10 mL) and H2O (2.5 mL). The reaction mixture was removed from the ice bath and stirred at room temperature. After 1.5 hours, the reaction mixture was concentrated under reduced pressure at 37°C. The resulting residue was dissolved in dichloromethane (10 mL), cooled to 0°C, and then TFA (5 mL) was added. After 1.5 hours, the reaction mixture was concentrated under reduced pressure at 37°C. The resulting residue was diluted with DCM (2 mL) and toluene (2 mL) and then concentrated under reduced pressure. The resulting brown oily substance was dissolved in DMF (5 mL), cooled to 0°C, and then DIPEA (1.3 mL, 7.65 mmol, 5 equivalents), dipropargylamine (0.8 mL, 7.65 mmol, 5 equivalents), and HATU (1.098 g, 2.89 mmol, 1.9 equivalents) were added in sequence. The reaction mixture was removed from the ice bath and stirred at room temperature. After 75 minutes, the reaction mixture was diluted with ethyl acetate-hexane (9:1, 50 mL) and 1 N HCl (50 mL), and stirred vigorously for 5 minutes. The aqueous layer was extracted with ethyl acetate-hexane (9:1, 3 × 50 mL). The combined organic extracts were washed with water (2 × 50 mL) and saturated sodium chloride aqueous solution (50 mL). The aqueous layer was separately extracted with ethyl acetate-hexane (4:1, 50 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting yellow solid (1.62 g) was suspended in dichloromethane (approximately 10 mL) and filtered through plastic frit. The filter cake was washed with DCM (2 × 10 mL), and the combined filtrate was concentrated under reduced pressure. The resulting residue (1 g) was purified by flash column chromatography on silica gel (25 g, gradient: ethyl acetate in hexane, 0% [2 CV] → 60% [5 CV], 60% [2 CV], 60 → 90% [3 CV], 90% [4 CV], 90 → 95% [3 CV], 95% [4 CV]) to obtain amide 7 (532 mg, 84%) as an off-white foam. LCMS: ESI+ m / z 438.04[M+Na] + .

[0479] [ka] The stock solution and the water and THF for the reaction were sparged with argon for 15 minutes via a needle from a balloon. Tris-alkyne (780 mg, 1.88 mmol, 1 equivalent) and azide (3.2 g, 5.9 mmol, 3.2 equivalents) were placed under an argon atmosphere, dissolved in THF (18 mL), and sparged with argon for 15 minutes. Aqueous solutions of sodium ascorbate and copper sulfate (twice the mass and volume required for the reaction) were prepared in scintillation vials and sparged with argon for 15 minutes. A solution of sodium ascorbate (151 mg, 0.76 mmol, 0.4 equivalents) in water (4.5 mL) was added, followed by a solution of copper sulfate pentahydrate (94 mg, 0.38 mmol, 0.2 equivalents) in water (4.5 mL), which was added dropwise to the THF solution of alkyne and azide over 5 minutes at 0°C. After 5 minutes, the reaction mixture was removed from the ice bath and allowed to rise to room temperature. After 2.5 hours, the reaction mixture was concentrated under reduced pressure (50 torr) at 35°C to remove THF. The reaction mixture was diluted with DCM (100 mL) and washed with 50% saturated sodium bicarbonate aqueous solution (100 mL), water (100 mL), and saturated sodium chloride aqueous solution (100 mL). The aqueous layer was separately extracted with dichloromethane (2 × 50 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting off-white foamy substance (total 2.8 g) was re-purified by flash column chromatography on silica gel (25 g, high-capacity 20 μm cartridge, gradient: methanol in dichloromethane, 0% [2 CV], 0 → 10% [4 CV], 10 → 15% [8 CV], 15% [13 CV], 15 → 20% [5 CV], 20% [10 CV]) to obtain click-reacted product 8 (1.86 g, 96% HPLC purity, 48% yield) and a lower purity batch (494 mg, 90% purity, 12% yield). LCMS: ESI+ m / z 2082.8 [M+Na] + .

[0480] [ka] Alcohol 8 (1.86 g, 0.903 mmol, 1 equivalent) was dried azeotropically by concentration in a rotavap at 35°C under reduced pressure, with argon backfilling the flask through the Schlenk port, from anhydrous DCM (10 mL) and toluene (15 mL). The DCM for the reaction was sparged in bulk with argon for 15 minutes. Alcohol 8 was dissolved in anhydrous DCM (30 mL), then DIPEA (1.1 mL, 6.3 mmol, 7 equivalents) was added, followed by 2-cyanoethyl=N,N-diisopropylchlorophosphorus. Roamidite (0.60 mL, 2.7 mmol, 3 equivalents) was added dropwise. HPLC analysis at 10 minutes showed complete conversion. After 25 minutes, the reaction mixture was quenched by direct addition of 50 mL of 50% saturated sodium bicarbonate aqueous solution. The aqueous layer was extracted with DCM (2 × 50 mL). The combined organic extract was washed with 50 mL of 10% saturated sodium bicarbonate aqueous solution, with 20 mL of saturated sodium chloride solution added to aid in emulsion formation and disruption, and 50 mL of saturated sodium chloride aqueous solution. The aqueous layer was extracted separately with DCM (2 × 50 mL). The combined organic extract was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained white foamy substance (3.2 g) was purified by flash column chromatography on silica gel (25 g high-capacity 20 μm column, 3 CV, pre-equilibriumized with 2% triethylamine in dichloromethane, gradient: constant 2% triethylamine, methanol in dichloromethane, 0% [2 CV], 0 → 4% [8 CV], 4 → 5% [5 CV], 5 → 7% [5 CV], 7 → 10% [5 CV], 10% [2 CV]) to obtain amidite. The fractions were analyzed for purity by HPLC: f14 (96%), f15 (94.5%), f16 (94%), f17 (91%), f18 (88%), f19 (86%), f22 (34%). Combining fractions f14-17, amidite 9 (2.00g, HPLC purity 93% → 1.86g, yield 91%) was obtained as a white foamy substance (1H and 31P NMR were clear - DMSO-d6).

[0481] [ka] Step 1: SEMCl (0.87 mL, 4.9 mmol, 1.2 equivalents) was added dropwise at 0°C to a solution of disiloxane (2.00 g, 4.11 mmol, 1 equivalent) and DBU (0.92 mL, 6.2 mmol, 1.5 equivalents) in anhydrous DMF (21 mL, 0.2 M). The flask was placed in an ice bath and slowly heated. After 17 hours, the reaction mixture was poured into a mixture of stirred ethyl acetate-hexane (5:1, 200 mL) and cold water (200 mL). The organic layer was washed with water (200 mL) and saturated sodium chloride aqueous solution (150 mL). The aqueous layer was then separated into ethyl acetate-hexane (5:1, 200 mL) The organic extracts were extracted once using ) . The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.

[0482] Step 2: The obtained residue was dried by azeotropy after concentration from anhydrous toluene (20 mL). The obtained residue was dissolved in DMF (21 mL), cooled to 0°C, and then t-butyl bromoacetate (1.52 mL, 10.3 mmol, 2.5 equivalents) and sodium hydride (60% dispersion in mineral oil, 329 mg, 8.2 mmol, 2 equivalents) were added. The reaction mixture was placed in an ice bath and slowly heated. After 2 hours, the reaction mixture was quenched with methanol (0.5 mL) and immediately poured into a stirring mixture of ethyl acetate-hexane (4:1, 150 mL) and a 10% saturated sodium chloride aqueous solution (200 mL). The organic layer was washed with a 10% saturated sodium chloride aqueous solution (2 × 100 mL) and a saturated sodium chloride aqueous solution (100 mL). The aqueous layer was extracted once separately with ethyl acetate-hexane (4:1, 100 mL). The combined organic extracts were dried with anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure.

[0483] Step 3: The obtained residue was dissolved in THF (25 mL), cooled to 0°C, then TBAF (1.0 M, 10 mL, 10 mmol, 2.5 equivalents in THF) was added, and the ice bath was removed. After 30 minutes, the reaction mixture was poured into a stirred saturated sodium bicarbonate aqueous solution (100 mL) and DCM (100 mL). The aqueous layer was extracted with DCM (2 × 100 mL). The final organic layer was set aside for later extraction. The combined first two organic extracts were washed with water (100 mL) and saturated sodium chloride aqueous solution (100 mL). The aqueous layer was separately extracted with the reserved DCM layer. The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure.

[0484] Step 4: The obtained residue was dried by azeotropy after concentration from anhydrous toluene (20 mL). The obtained residue was dissolved in DMF (15 mL) and cooled to 0°C, after which imidazole (559 mg, 8.2 mmol, 2 equivalents) and TBSCl (619 mg, 4.1 mmol, 1 equivalent) were added. The ice bath was removed and the reaction mixture was stirred at room temperature. After 1.5 hours, further imidazole (303 mg, 4.5 mmol, 1 equivalent) and TBSCl (325 mg, 2.2 mmol, 0.5 equivalents) were added. After another 30 minutes, this reaction mixture was poured into stirred ethyl acetate-hexane (5:1, 150 mL) and cold water (150 mL). The organic layer was washed with water (2 × 150 mL) and saturated sodium chloride aqueous solution (150 mL). The aqueous layer was extracted once separately with ethyl acetate-hexane (5:1, 100 mL). The combined organic extracts were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue (5 g) was purified by flash column chromatography using silica gel (50 g, 60 μm, ethyl acetate in hexane, 0% [2 CV], 0 → 8% [5 CV], 8 → 15% [8 CV], 15% [7 CV], 15 → 30% [5 CV], 30% [5 CV]) to obtain alcohol (2.25 g, 90%) as a colorless oil.

[0485] Example 10 [ka]

[0486] [ka] To a suspension of commercially available GalNAc acetate (11.5 g, 29.579 mmol, 1 equivalent) in 1,2-dichloroethane (70 mL), TMSOTf (6.4 mL, 35.495 mmol, 1.2 equivalents, d: 1.228) was added over 5 minutes at room temperature, and then this The mixture was heated to 50°C and stirred at 50°C for 5 hours. The mixture was allowed to cool to room temperature, 20 g of 4A MS was added, followed by 10.0 g of HO-PEG 6 N3 (32.537 mmol, 1.1 equivalents). After stirring at room temperature for 10 minutes, TMSOTf (1.8 mL, 9.762 mmol, 0.3 equivalents) was added. The resulting mixture was stirred at room temperature for 2 days. Workup: The reaction mixture was filtered and washed with DCM (50 mL). The filtrate was poured into brine (100 mL) + Sat NaHCO3 (100 mL). The organic layer was separated, and the aqueous phase was extracted with DCM (50 mL). The combined organic layers were washed with brine (100 mL) + Sat NaHCO3 (50 mL) and concentrated. The residue was purified by flash chromatography using 50% acetone / hept, then acetone, and then 1% to 6% MeOH / acetone to obtain azide 11 (10.4 70%) as a pale yellow syrup.

[0487] [ka] To a solution of compound 9 (1.4 g, 2.428 mmol, 1 equivalent) and azide 11 (5.1 g, 8.013 mmol, 3.3 equivalents) in anhydrous THF (28 mL), 243 mg (0.971 mmol, 0.4 equivalents) of CuSO4.5H2O (7 mL, 0.4 equivalents) was added at 0°C, followed by the addition of L-Na-ascorbate (1.457 mmol, 0.6 equivalents) in 7 mL of water, and then the cooling bath was removed. The resulting mixture (pale yellow) was stirred at room temperature for 5 hours. Workup: NH4Cl-NH3.H2O (preparation: 5 g of NH4Cl, 5 mL of NH3.H2O (29%) in 50 mL of water) (20 mL) was added, and the aqueous phase was extracted using DCM (50 mL x 4). The combined organic layers were washed twice with brine (100 mL) + NH4Cl-NH3.H2O (10 mL), then washed with brine (80 mL) + NH3.H2O (1 mL) until the aqueous phase no longer showed blue color, and then washed with brine (80 mL). The residue was concentrated and purified by flash chromatography with 50% siRNA / Hept, then siRNA, and then 1%-4% MeOH / DCM to obtain product 10 as a white foam. 4.9 g, yield 81%.

[0488] [ka] A stirred solution of alcohol 10 (2.93 g, 1.179 mmol, 1 equivalent) and diisopropylethylamine (1.23 mL, 7.072 mmol, 6 equivalents) in DCM (30 mL) is mixed with 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite (0. 78 mL (3.536 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour, and the reaction mixture was quenched with saturated NaHCO3 aqueous solution (50 mL) and DCM was used. Extraction was performed with (100) ml of brine, washed with brine (100 mL), dried over N2SO4, and evaporated. The crude product was loaded onto a Biotage silica gel column (50 g 20 μm) (pre-equilibrated twice with 1% Et3N-DCM) and purified by flash chromatography using 0-10% MeOH / DCM with 1% Et3N as an additive. The pure fractions were combined, concentrated, and dried under high vacuum to obtain phosphoramidite 12 (2.8 g, 88%) as a beige solid. Purity was 92% by HPLC, and P31-NMR, mass (m / z 2708 M+Na), and H1-NMR corresponded to the product.

[0489] Example 11 [ka]

[0490] [ka] A stirred solution of alkyne 7 (0.65 g, 1.566 mmol, 1 equivalent) and azide 11 (3.18 g, 5.012 mmol, 3.2 equivalents) in THF (30 mL) is mixed with water (5 mL). CuSO4·5H2O (0.19 g, 0.788 mmol) in L) and sodium ascorbate (0.23 g, 1.198 mmol) in water (5 mL) were added, and the mixture was stirred at room temperature for 3 hours. LC-MS showed product formation and complete disappearance of the alkyne. This reaction mixture was diluted with aq. saturated NaHCO3 (50 mL), extracted with 2 × 50 mL of DCM, and washed with brine solution (50 mL). The combined extracts were dried over Na2SO4 and concentrated. The resulting crude product was purified by column chromatography using 0-30% MeOH / DCM as eluate. The pure fractions were combined and concentrated, and the resulting solid was co-evaporated with toluene and dried under high vacuum to obtain tris-triazole (2.8 g 76%) as a white solid. LC-MS and NMR correspond to the product. [ka]

[0491] A stirred solution of alcohol 12 (1.033 g, 0.444 mmol, 1 equivalent) and diisopropylethylamine (0.46 mL, 2.666 mmol, 6 equivalents) in DCM (10 mL) is mixed with 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite (0 0.29 mL (1.33 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (50 mL). The crude product was extracted with DCM (100 mL), washed with brine (100 mL), dried over Na2SO4, and evaporated. The crude product was loaded onto a Biotage silica gel column (25 g 20 μm) (pre-equilibrated with 1% Et3N-DCM) and purified by flash chromatography using 0-10% MeOH / DCM containing 1% Et3N as an additive. The pure fractions were combined, concentrated, and dried under high vacuum to obtain phosphoramidite 13 (1.01 g, 90%) as a beige solid. Purity was 91% by HPLC, and P31-NMR, mass (m / z 2546 M-1+ Na), and H1-NMR corresponded to the product.

[0492] Example 12 [ka]

[0493] [ka] To a stirred solution of alcohol 1 (0.85 g, 1.974 mmol, 1 equivalent) in anhydrous pyridine (10 mL), DMTrCl (0.87 g, 2.566 mmol, 1.3 equivalents) was added. The reaction mixture was stirred overnight at room temperature. The reaction was quenched by adding MeOH. The crude reaction mixture was concentrated under reduced pressure. The residue was partitioned between SiO2 and NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by biotage column (25 g, 20 μM) using 0-60% SiO2 in hexane (at 10 CV) to obtain 0.96 g of product 2 (68%), a slightly yellow solid. It was obtained as such.

[0494] [ka] Compound 2 (0.8 g, 1.115 mmol, 1 equivalent) was stirred in anhydrous CH3CN (10 mL) and DMAP (272 mg, 2.229 mmol, 2 equivalents), triethylamine (0.311 mL, 2.229 mmol, 2 equivalents), and TPSCl (675 mg, 2.229 mmol, 2 equivalents) were added. The reaction mixture was stirred at room temperature for 2 hours. Concentrated ammonium hydroxide (50% v / v in H2O) (5 mL) was added to the reaction mixture. The reaction mixture was stirred overnight. The crude reaction mixture was concentrated under reduced pressure. The residue was partitioned between DCM and sat.NH4Cl. The aqueous layer was extracted with DCM (once). The organic layer was collected, dried over Na2SO4, filtered, and concentrated. The residue was purified using a biotage column (25g, 20uM) with 0-8% MeOH in DCM (Diamond-Compound Cells) in a 10CV column to obtain 661mg of product 3 (83%) as an off-white solid.

[0495] [ka] To a solution of amine 3 (660 mg, 0.921 mmol, 1 equivalent) in anhydrous DMF (5 mL), acetic anhydride (0.113 mL, 1.197 mmol, 1.3 equivalents) was added. The reaction mixture was stirred at room temperature for 60 hours. The reaction mixture was diluted with DCM and washed once with NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel chromatography (25 g, 20 μm) using 5 CV of 0-10% MeOH / DCM. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 550 mg of product 3 (81%) as a white solid.

[0496] [ka] To a solution of DMT ether 4 (0.61 g, 0.804 mmol, 1.00 equivalent) in DCM (5 mL), TFA (0.185 mL, 2.4 mmol, 3.0 equivalent) was added at room temperature. The reaction mixture was stirred at room temperature for 60 minutes. A further 0.1 mL of TFA was added to the reaction mixture, and the mixture was stirred for 30 minutes. This reaction was then completed by adding aq. NaHCO3. The reaction mixture was carefully quenched. The reaction mixture was partitioned between saturated NaHCO3 and DCM. The aqueous phase was extracted with DCM (once). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0-10% MeOH in DCM to obtain 235 mg of product 5 (64%) as a white solid.

[0497] [ka] To a solution of compound 5 (0.22 g, 0.482 mmol, 1 equivalent) and azide 12 (0.846 g, 1.542 mmol, 3.2 equivalents) in THF (4 mL), a solution of CuSO4·5H2O (0.036 g, 0.145 mmol, 0.3 equivalents) in water (1 mL) was added at 0°C, followed by a solution of sodium ascorbate (0.043 g, 0.217 mmol, 0.45 equivalents) in water (1 mL). After stirring at 0°C for 5 minutes, the reaction mixture was warmed to room temperature. After stirring the reaction mixture at room temperature for 2 hours, LC-MS showed the main product, including some double-clicked products (m / z: 777). Further azide 2 (0.132 g, 0.241 mmol, 0.5 equivalents) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 2 hours. LC-MS indicated completion of the reaction. The reaction product was diluted with DCM and washed with NaHCO3. The aqueous phase was extracted with DCM (twice). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified with 0-20% MeOH in DCM (25 g, 20 μM, 15 CV) to obtain 0.85 g of triazole 6 (84%) as a white solid.

[0498] [ka] A stirred solution of triazole 6 (0.78 g, 0.371 mmol, 1 equivalent) and diisopropylethylamine (0.42 mL, 2.412 mmol, 6.5 equivalents) in anhydrous DCM (15 mL) was prepared by adding 2-cyanoethyl=N,N-diisopropylchlorophosphorumid 0.248 mL (1.113 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 40 minutes. LC-MS and HPLC indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 solution and partitioned between DCM and saturated NaHCO3. The DCM phase was collected. The aqueous phase was extracted with DCM (once). The combined organic phases were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was loaded onto a pre-equilibrated (1% Et3N-DCM) biotage silica gel column (25 g, 20 μm) and 0-100% Et3N was added as an additive. The phosphoramidite was purified by flash chromatography using 5% MeOH / DCM (at 15 CV). The pure fractions were combined, concentrated, and dried under high vacuum to obtain 0.60 g of phosphoramidite 7 (98% purity according to HPLC) as an off-white solid. MS: m / z = 2323.7 [M + Na] + The P31-NMR and H1-NMR spectra correspond to the product.

[0499] Example 13 [ka]

[0500] [ka] Compound 1 (2.0 g, 2.275 mmol, 1 equivalent) was dissolved in 7N NH3MeOH (20 mL). The reaction mixture was stirred at room temperature for 2 hours. LC-MS indicated that the reaction was complete. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (50 g, 20 μm) using 0-10% MeOH / DCM at 15 CV. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 1.82 g of Product 2 as a white solid. Compound 2 (1.82 g, 2.349 mmol, 1 equivalent) was stirred in anhydrous CH3CN (20 mL) and DMAP (574 mg, 4.698 mmol, 2 equivalents), triethylamine (0.655 mL, 4.698 mmol, 2 equivalents), and 2,4,6-triisopropylbenzenesulfonyl chloride (1.423 g, 0.468 mmol, 2 equivalents) were added. The reaction mixture was stirred at room temperature for 45 minutes. Concentrated ammonium hydroxide (50% v / v in H2O) (5 mL) was added to the reaction mixture. The reaction mixture was stirred overnight. The crude reaction mixture was concentrated under reduced pressure. The residue was partitioned between DCM and sat.NH4Cl. The aqueous layer was extracted with DCM (once). The organic layer was collected, dried over Na2SO4, filtered, and concentrated. The residue was purified using a biotage column (50g, 20uM) with 0-10% MeOH in DCM at 15CV, yielding 1.75g ​​of product 3 (99%) as a white solid.

[0501] [ka] To a solution of compound 3 (1.75 g, 2.261 mmol, 1 equivalent) in anhydrous DMF (10 mL), acetic anhydride (0.278 mL, 2.94 mmol, 1.3 equivalents) was added. The reaction mixture was stirred at room temperature for 60 hours. LC-MS showed that a small amount of starting material (SM) remained. The reaction mixture was diluted with DCM and washed once with NaHCO3. The organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel chromatography (50 g, 20 μm) using 0-10% MeOH / siRNA at 10 CV. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 1.38 g of product 4 (75%) as a white solid.

[0502] [ka] To a solution of DMT ether 4 (1.4 g, 1.716 mmol, 1.00 equivalent) in DCM (15 mL), TFA (0.394 mL, 5.148 mmol, 3.0 equivalent) was added at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. A further 0.2 mL of TFA was added to the reaction mixture, and the mixture was stirred for another 30 minutes. The reaction was carefully quenched by the addition of aq. NaHCO3. The reaction mixture was partitioned between saturated NaHCO3 and DCM. The aqueous phase was extracted with DCM (once). The organic layers were combined. The white insoluble solid in the organic layers was collected by filtration. LC-MS showed that this insoluble solid was product 5. The solid was dried under reduced pressure to obtain 586 mg of product 5 (67%) as a white solid.

[0503] [ka] To a suspension of compound 12 (0.41 g, 0.798 mmol, 1 equivalent) and azide 6 (1.445 g, 2.635 mmol, 3.3 equivalents) in THF (12 mL), a solution of CuSO4·5H2O (0.060 g, 0.240 mmol, 0.3 equivalents) in water (3 mL) was added at room temperature, followed by the addition of a solution of sodium ascorbate (0.071 g, 0.359 mmol, 0.45 equivalents) in water (3 mL). The reaction mixture was stirred at room temperature for 7 hours. The reaction mixture was then diluted with DCM and washed with NaHCO3. The aqueous phase was extracted with DCM (once). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified with 0-20% MeOH (50 g, 20 uM, 15 CV) in DCM to obtain 1.24 g of triazole 7 (71%) as a white solid.

[0504] [ka] A stirred solution of triazole 7 (1.63 g, 0.755 mmol, 1 equivalent) and diisopropylethylamine (0.855 mL, 4.9 mmol, 6.5 equivalents) in anhydrous DCM (20 mL) is prepared by adding 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite. (0.505 mL, 2.265 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 40 minutes. LC-MS and HPLC indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 solution and partitioned between DCM and saturated NaHCO3. The DCM phase was collected. The aqueous phase was extracted with DCM (once). The combined organic phase was washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was loaded onto a pre-equilibrated (1% Et3N-DCM) biotage silica gel column (50 g, 20 μm) and purified by flash chromatography using 0-15% MeOH / DCM (at 15 CV) with 1% Et3N as an additive. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 1.3 g of phosphoramidite 8 (98% purity according to HPLC) as an off-white solid. MS:m / z=2381.5[M+Na] + The P31-NMR and H1-NMR spectra correspond to the product.

[0505] Example 14 [ka]

[0506] [ka] To a stirred, cooled (0°C) solution of Compound 1 (10.08 g, 18.462 mmol, 1 equivalent) in anhydrous DMF (100 ml), NaH 60 w% (2.58 mg, 64.615 mmol, 3 equivalents) was added. The reaction mixture was stirred at 0°C for 30 minutes. Propargyl bromide (8.236 mL, 55.38 mmol, 3 equivalents) was added dropwise, and stirring was continued at 0°C for 2 hours. The reaction mixture was quenched with water (200 mL), extracted with ethyl acetate (200 mL), washed with water (2 × 100 mL) and brine (100 mL), dried (Na₂SO₄), concentrated, and the residue was purified by silica gel column chromatography using 0–70% ethyl acetate / hexane as the eluate. The pure fractions were combined and concentrated to obtain the tri-propargyl product as a white solid (11 g, 90%). The product was identified by NMR and LC-MS (m / z 684 M+Na).

[0507] [ka] TFA (2.4 mL, 31.384 mmol, 5 equivalents) was added to a stirred, cooled 0°C solution of DMT ether (10 g, 14.265 mmol, 1.00 equivalent) in DCM (50 mL). The reaction mixture was stirred at room temperature for 3 hours. LC-MS showed complete deprotection. The reaction mixture was quenched with NaHCO3 (50 mL), extracted with DCM (2 × 200 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0–100 toluene / hexane as eluate. The pure fractions were combined and concentrated to obtain alcohol (4.5 g, 83%) as a white solid. The product was confirmed by NMR and LC-MS. M / z 359 (M+1).

[0508] [ka] To a stirred solution of Tris-alkyne 3 (1.07 g, 2.98 mmol, 1 equivalent) and azide 11 (6.08 g, 9.56 mmol, 3.2 equivalents) in THF (20 mL), CuSO4·5H2O (0.37 g, 1.49 mmol, 0.5 equivalents) in water (2 mL) was added, followed by sodium ascorbate (0.44 g, 2.24 mmol, 0.75 equivalents) in water (2 mL), and the mixture was stirred at room temperature for 3 hours. LC-MS showed complete disappearance of the alkyne. This reaction mixture was diluted with aq. saturated NaHCO3 (50 mL) and extracted with DCM (2 × 50 mL) and 10% DCM:MeOH (50 mL). The combined organic layers were washed with 100 mL of brine solution, dried over Na2SO4, and concentrated. The crude product was purified by column chromatography using 0-40% MeOH / DCM as the eluent. The pure fractions were combined and concentrated, and the resulting solid was co-evaporated with toluene and dried under high vacuum to obtain the tris-triazole product (5 g; 72%) as a white solid. LC-MS and NMR correspond to the product.

[0509] [ka] A stirred solution of alcohol 5 (0.6 g, 0.724 mmol, 1 equivalent) and diisopropylethylamine (0.199 g, 4.346 mmol, 6 equivalents) in DCM (20 mL) is mixed with 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite (0.18 (g, 0.77 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 30 minutes. LC-MS indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (20 mL), extracted with DCM (2 × 30 mL), washed with brine (20 mL), dried over N2SO4, and evaporated. The crude product was loaded onto a pre-equilibriumized (1% Et3N-MeOH:1% DCM (1:10)) Biotage silica gel column (50 g 20 μm) and purified by flash chromatography using 0–10% MeOH / DCM with 1% Et3N as an additive. The pure fraction was collected and concentrated to obtain 0.5 g (77%) of product as a white solid with >95% HPLC purity. Mass and NMR correspond to the product.

[0510] Example 15 [ka]

[0511] [ka] Benzoyl derivative 8 (7.3 g, 8.314 mmol, 1 equivalent) was dissolved in 7N NH3-MeOH (70 mL). The reaction mixture was stirred at room temperature for 2 hours. LC-MS indicated completion of the reaction. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel chromatography (100 g, 60 μm) using 0-10% MeOH / DCM at 15 CV. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 6.2 g. The product (96%) was obtained as a white solid. LC-MS m / z=775 M+Na and NMR correspond to the product.

[0512] [ka] To a stirred solution of uridine derivative 9 (6.15 g, 7.946 mmol, 1 equivalent) in anhydrous CH3CN (60 mL), DMAP (1.94 g, 15.891 mmol, 2 equivalents), triethylamine (2.215 mL, 15.891 mmol, 2 equivalents), and 2,4,6-triisopropylbenzenesulfonyl chloride (4.813 g, 15.891 mmol, 2 equivalents) were added. The reaction mixture was stirred at room temperature for 1 hour. LC-MS showed consumption of the starting materials. Concentrated ammonium hydroxide (50% v / v in H2O) (15 mL) was added to the reaction mixture, and the mixture was stirred overnight. The crude reaction mixture was concentrated under reduced pressure. The residue was partitioned between DCM and sat.NH4Cl. The aqueous layer was extracted with DCM (1 × 200 mL). The organic layer was collected, dried over Na2SO4, filtered, and concentrated. The residue was purified using a Biotage column (50 g, 20 μM) with 0-10% MeOH in DCM at 15 CV, yielding 5.2 g of product (85%) as a white solid. The NMR and LCMS m / z 774 (M+1) values ​​correspond to the product.

[0513] [ka] To a solution of amine 10 (5.12 g, 6.624 mmol, 1 equivalent) in anhydrous DMF (40 mL), acetic anhydride (0.75 mL, 7.948 mmol, 1.2 equivalents) was added. The reaction mixture was stirred at room temperature for 60 hours. The reaction mixture was diluted with DCM (300 mL) and washed with NaHCO3 (100 mL). The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (50 g, 20 μm) using 0-10% MeOH / siRNA at 10 CV. The pure fractions were combined, concentrated, and dried under high vacuum to obtain 4.4 g of acetate (82%) as a white solid. NMR and LCMS m / z 816 (M+1) correspond to the product.

[0514] [ka] To a solution of DMT ether 12 (4.35 g, 5.337 mmol, 1.00 equivalent) in DCM (40 mL), TFA (1.63 mL, 21.35 mmol, 4.0 equivalent) was added at room temperature, and the mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with DCM (100 mL) and carefully quenched by adding aq. NaHCO3 (100 mL). The product precipitated as a white solid. The solid was collected by filtration, washed with DCM, and dried under high vacuum to obtain 2.25 g of the product (82%) as a white solid. NMR and LCMS m / z=514 (M+Na) correspond to the product.

[0515] [ka] To a suspension of compound tri-alkyne 13 (2.2 g, 4.288 mmol, 1 equivalent) and azide 11 (9 g, 14.152 mmol, 3.3 equivalents) in THF (50 mL), a solution of CuSO4·5H2O (535 mg, 2.144 mmol, 0.5 equivalents) in water (10 mL) was added at room temperature, followed by the addition of a solution of sodium ascorbate (198 mg, 3.216 mmol, 0.75 equivalents) in water (10 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was then diluted with DCM and washed with NaHCO3 (100 mL). The aqueous phase was extracted with DCM (2 × 500 mL). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified with 0-20% MeOH (100g, 20uM, 15CV) in DCM to obtain 8.31g of triazole (80%) as a white solid. The NMR and mass m / z = 2445 (M + Na) correspond to the product.

[0516] [ka] A stirred solution of alcohol 14 (6.00 g, 2.474 mmol, 1 equivalent) and diisopropylethylamine (2.58 mL, 14.858 mmol, 6 equivalents) in DCM (50 mL) is mixed with 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite (1 0.65 mL (7.429 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (100 mL), extracted with DCM (500 mL), washed with brine (100 mL), dried over N2SO4, and evaporated. The crude product was loaded onto a Biotage silica gel column (100 g 20 μm) (pre-equilibrated twice with 1% Et3N-DCM) and purified by flash chromatography using 0-10% MeOH / DCM containing 1% Et3N as an additive. The pure fractions were combined, concentrated, and dried under high vacuum to obtain phosphoramidite 15 (6.9 g). NMR showed the triethylamine salt. This compound was dissolved in 150 ml of DCM, washed with 2 × 50 mL of saturated bicarbonate aqueous solution, and the organic layer was dried over Na₂SO₄, evaporated, and dried under high vacuum to obtain a (5.9 g 91%) product as a white solid. HPLC showed a purity of 95%, and the P31-NMR, mass (m / z 2644 M+Na), and H1-NMR corresponded to the product.

[0517] Example 16 [ka]

[0518] [ka] To a solution of starting material 1 (5.0 g, 10.27 mmol, 1 equivalent) in anhydrous pyridine (40 mL), chlorotrimethylsilane (3.129 mL, 24.655 mmol, 2.4 equivalents) was added dropwise at room temperature. This mixture was stirred for 2 hours, and then benzoyl chloride ( 2.862 mL (24.655 mmol, 2.4 equivalents) was added dropwise to the reaction mixture. The reaction mixture was stirred overnight at room temperature. 10 mL of H₂O was added to the reaction mixture, and the mixture was stirred for 8 hours. The solvent was removed by rotavap. The crude mixture was partitioned between H₂O and SiO₂. The aqueous phase was extracted with SiO₂ (2 × 100 mL). The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated. The residue was purified with 0–30% SiO₂ in hexane to obtain 5.2 g of Product 2 (85%) as a white solid.

[0519] [ka] Dipropargylamine (5 g, 53.688 mmol, 1 equivalent) and K2CO3 (37.1 g, 268.44 mmol, 5 equivalents) were suspended in CH3CN (50 mL), to which bromoacetyl chloride (4.471 mL, 53.688 mmol, 1 equivalent) was added at 0°C. The reaction mixture was stirred at 0°C for 1 hour. LC-MS indicated that the reaction was complete. The reaction mixture was diluted with DCM and washed with water. The organic phase was dried over Na2SO4, concentrated, and purified by column chromatography (0% → 30% toluene-hexane) to obtain 9 g of Product 3 (58%) as a yellow oil.

[0520] [ka] To a stirred solution of alcohol 2 (7.1 g, 12.017 mmol, 1 equivalent) in DMF (25 mL), amide 3 (3.86 g, 18.026 mmol, 1.5 equivalents) was added at 0°C, followed by the addition of NaH (0.625 g, 60% of mineral oil, 15.622 mmol, 1.3 equivalents) in two separate additions. The reaction mixture was stirred at 0°C for 45 minutes, then quenched by adding ice water. The precipitated solid was filtered, redissolved in DCM, dried over Na2SO4, and concentrated. This crude product was used directly in the next step.

[0521] [ka] To a stirred solution of bis-alkyne 4 (7.2 g, 9.945 mmol, 1 equivalent) in THF (100 mL), TBAF (5.19 g, 19.917 mmol, 2 equivalents) was added. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with SiO2 and washed with NaHCO3 and brine. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was flash-chlorous acid using 0-5% MeOH in SiO2. The product was purified by matrixing to obtain 4.5 g of product 5 (79% in two steps) as a white solid.

[0522] [ka] To a stirred solution of diol 5 (11 g, 18 mmol, 1.00 equivalent) in DCM (100 mL), TEA (3.7 g, 36.9 mmol, 2 equivalents) and DMAP (0.27 g, 2.2 mmol, 0.1 equivalent) were added, followed by the addition of TBDMS-Cl (3.3 g, 22.1 mmol, 1.2 equivalents). This reaction mixture was stirred at room temperature for 12 hours. LC-MS showed a complete reaction. Water (50 mL) was added to the reaction mixture, and the DCM layer was separated, dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0-60% hexane / EA as the eluent, and the pure fractions were combined and concentrated to obtain the product as a white solid (11.5 g; 84%). The product was confirmed by LC-MS.

[0523] [ka] To a stirred mixture of alcohol 6 (11.5 g, 19.3 mmol, 1 equivalent) and bromo compound 7 (4.4 g, 23.19 mmol, 1.3 equivalents) in THF, NaH (0.55 g, 23.19 mmol, 1.2 equivalents) was added at -10°C. This reactant (RM) was stirred at -10°C for 1 hour. After completion, the reaction was quenched with water, extracted with EA, dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude product was purified by Biotage column chromatography (0-60% ethyl acetate / hexane) to obtain compound 8 as a beige solid (7 g, 52%). LC-MS and 1 The 1H-NMR spectrum corresponds to the product.

[0524] [ka] 50 ml of an aqueous solution of NH4OH was added to a solution of compound 8 (6.8 g, 9.8 mmol, 1 equivalent) in THF (50 mL), and the mixture was stirred at room temperature for 3 hours. After the reaction was complete, water was added, and the mixture was extracted with DCM, dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting crude product was purified by Biotage column chromatography (1:10 DCM / MeOH) to obtain compound 3 as a beige solid (4 g, 70%).

[0525] [ka] To a stirred solution of uridine derivative 9 (4.5 g, 7.143 mmol, 1 equivalent) in anhydrous CH3CN (50 mL), DMAP (462 mg, 3.784 mmol, 2 equivalents), triethylamine (0.527 mL, 3.784 mmol, 2 equivalents), and TiPSCl (1.146 g, 2.3784 mmol, 2 equivalents) were added. The reaction mixture was stirred at room temperature for 2 hours. Concentrated ammonium hydroxide (50% v / v in H2O) (10 mL) was added to the reaction mixture. The reaction mixture was stirred overnight. The crude reaction mixture was concentrated under reduced pressure. The residue was partitioned between DCM and sat.NH4Cl. The aqueous layer was extracted with DCM (1 × 100 mL). The organic layer was collected, dried over Na2SO4, filtered, and concentrated. The residue was purified using a Biotage column (25g, 20uM) with 0-5% MeOH in DCM at 10CV, yielding 3.4g of cytosine (91%) as an off-white solid.

[0526] [ka] Compound 9 (4 g, 6.83 mmol, 1 equivalent) and benzoic anhydride (3.09 g, 13.67 mmol, 2 equivalents) were dissolved in 50 mL of dry DMF, and the reaction mixture was stirred at 25°C for 24 hours. Then, Py (2 ml) was added, and the reaction mixture was stirred at room temperature for a further 12 hours. After the completion of this reaction, excess DMF was removed under reduced pressure, and the crude product was purified using silica gel column chromatography (0-100% ethyl acetate / hexane) to obtain compound 12 as a beige solid (2.8 g 77%).

[0527] [ka] To a stirred solution of TBS-ether 12 (1.4 g, 2.031 mmol, 1 equivalent) in THF (30 mL), TBAF (1 g, 4.063 mmol, 2 equivalents) was added. The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was diluted with HCl and washed with NaHCO3 and brine. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified by flash chromatography using 0-10% MeOH in HCl to obtain 0.8 g of product 13 (70%) as a white solid.

[0528] [ka] To a stirred solution of alkyne 13 (0.8 g, 1.39 mmol, 1 equivalent) and azide 11 (2.83 g, 4.45 mmol, 3.2 equivalents) in THF (40 mL), CuSO4·5H2O (0.17 g, 0.64 mmol, 0.5 equivalents) in water (5 mL) was added, followed by sodium ascorbate (0.2 g, 1.04 mmol, 0.75 equivalents) in water (5 mL). This mixture was stirred at room temperature for 3 hours. LC-MS showed complete product formation and disappearance of the alkyne. This reaction mixture was diluted with aq. saturated NaHCO3 (50 mL) and extracted with DCM (2 × 50 mL). The organic layers were combined, washed with brine solution (100 mL), dried over Na2SO4, concentrated, and the resulting crude product was purified by column chromatography using 0–30% MeOH / DCM as the eluate. The pure fractions were combined and concentrated, and the resulting solid was co-evaporated with toluene and dried under high vacuum to obtain tris-triazole (2.7 g 78%) as a white solid. LC-MS and NMR correspond to this product.

[0529] [ka] A stirred solution of alcohol 14 (1.8g, 0.724 mmol, 1 equivalent) and diisopropylethylamine (0.56g, 4.346 mmol, 6 equivalents) in DCM (20mL) is mixed with 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite (0.51 (g, 2.173 mmol, 3 equivalents) was added dropwise. The reaction mixture was stirred at room temperature for 30 minutes. LC-MS indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (20 mL), extracted with DCM (2 × 30 mL), washed with brine (20 mL), dried over N2SO4, and evaporated. The crude product was loaded onto a pre-equilibriumized (1% Et3N-MeOH:1% DCM (1:10) Biotage silica gel column (50 g 20 μm) and purified by flash chromatography using 0-10% MeOH / DCM containing 1% Et3N as an additive. The pure fraction was collected and concentrated to obtain 1.5 g of Amidite 15 (80%) with >95% HPLC purity. NMR and mass correspond to this product.

[0530] Example 17 [ka]

[0531] [ka] 6-Chloroprin riboside (5 g, 17.483 mmol) and 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane compound (6.6 g, 20.978 mmol) were dissolved in anhydrous DMF (50 mL). TEA (7.25 mL, 52.447 mmol) was added to this reaction mixture, followed by DMAP (0.2 g, 1.75 mmol). The reaction mixture was stirred at room temperature for 6 hours. The reaction mixture was then poured into water (100 mL), and the resulting suspension was extracted with dichloromethane (3 × 50 mL). The combined organic extracts were washed with brine (1 × 20 mL), dried over magnesium sulfate, and concentrated under reduced pressure. The resulting crude product was purified by Biotage column chromatography (0-60% hexane / ethyl acetate) to obtain compound 2 (8 g, 86%) as a white solid.

[0532] [ka] To a stirred mixture of compound 2 (5 g, 9.452 mmol) and bromo compound 3 (2.42 g, 11.342 mmol) in DMF (50 mL), NaH (0.272 g, 11.342 mmol) was added at -10°C. The reactant was stirred at -10°C for 1 hour. After the reaction was complete, ice-cold water was added to the reactant. The precipitated solid was filtered, redissolved in DCM, dried over Na2SO4, filtered, and concentrated. The crude product was used in the next step without further purification.

[0533] [ka] To a stirred solution of di-silyl ether 4 (4 g, 6.042 mmol) in THF (50 mL), TBAF (3.13 g, 12.085 mmol) was added. The reaction mixture was stirred at room temperature for 3 hours. LC-MS showed complete deprotection. Water (50 mL) was added to the reaction mixture, extracted with DCM (2 × 50 mL), dried over Na₂SO₄, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0–10% DCM / MeOH as eluent, and the pure fractions were combined and concentrated to obtain diol 5 (2 g, 80%) as a white solid.

[0534] [ka] To a stirred solution of diol 5 (2 g, 4.773 mmol) in DCM (20 mL), TEA (0.96, 9.54 mmol) and DMAP (0.1 g) were added, followed by TBDMSCl (0.86 g, 5.728 mmol). The reaction mixture was stirred at room temperature for 12 hours. LC-MS indicated that the reaction was complete. The reaction mixture was quenched with water (50 mL), extracted with DCM (2 × 40 mL), dried over Na₂SO₄, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0–60% hexane / EA as the eluate, and the pure fractions were combined and concentrated to obtain product 6 (1.8 g, 75%) as a white solid.

[0535] [ka] To a stirred mixture of alcohol 6 (1.8 g, 3.371 mmol) and bromo compound 7 (0.71 g, 4.045 mmol) in DMF (20 mL), NaH (0.097 g, 4.045 mmol) was added at -10°C. The reaction mixture was stirred at -10°C for 1 hour. After completion, the reaction was quenched with water, extracted with EA, washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The resulting crude product was purified by Biotage column chromatography (hexane / ethyl acetate at 0-60°C) to obtain product 8 (1.3 g, 60%) as a white solid.

[0536] [ka] A mixture of 8 (0.1 g, 0.159 mmol), aq.NH3 (2 mL), and dioxane (5 mL) in a sealed tube was heated at 70°C for 12 hours. After the reaction was complete, excess dioxane and water were removed with rotavap. The resulting crude product was then mixed with 0-100 vinegar. The solution was purified by Biotage column chromatography using ethyl acid to obtain (0.075 g, 70%) as a beige solid.

[0537] [ka] BzCl (0.023 mL, 0.328 mmol) was added to a stirred solution of amine 9 (0.1 g, 0.164 mmol) in Py (5 mL). The reactants were stirred at 0°C for 2 hours. After the reaction was complete, 2N HCl was added, and the mixture was extracted with DCM. The organic layer was separated, dried over Na2SO4, filtered, and concentrated with rotavap. The resulting mixture of 10 and 10b was used directly in the next step.

[0538] [ka] To a stirred solution of crude 10 and 10b (0.15 g) in THF (5 mL), NH4OH (2 mL) was added. This reactant was stirred at 0°C for 3 hours. After the reaction was complete, 2N HCl was added to the reactant, and it was extracted with DCM. The organic layer was separated, dried over Na2SO4, filtered, and concentrated with rotavap. The resulting crude mixture was used in the next step without any purification.

[0539] [ka] A stirred solution of crude compound 12 (0.12 g) in THF (10 mL) is mixed with TBAF (0.1 g) was added. The reaction mixture was stirred at room temperature for 3 hours. LC-MS showed complete deprotection. Water (30 mL) was added to the reaction mixture, and then the reaction mixture was extracted with DCM (2 × 30 mL), dried over Na₂SO₄, filtered, and concentrated. The residue was purified by column chromatography on silica gel using 0-10% DCM / MeOH as the eluent, and the pure fractions were combined and concentrated to obtain (0.06 g 43%) as a white solid. The product was confirmed by LC-MS and NMR.

[0540] [ka] To a stirred solution of alkyne 13 (1.35 g, 2.25 mmol) and azide 11 (3.93 g, 7.21 mmol) in THF (50 mL), CuSO4·5H2O (0.28 g, 1.12 mmol) and sodium ascorbate (0.32 g, 1.62 mmol) in water (10 + 10 mL) were added, and the mixture was stirred at room temperature for 3 hours. LC-MS showed product formation and complete disappearance of the alkyne. This reaction mixture was diluted with aq. saturated NaHCO3 (100 mL), extracted with 2 × 100 mL of DCM, and the organic layer was washed with brine solution (100 mL). The combined extracts were dried over Na2SO4 and concentrated, and the resulting crude product was purified by column chromatography using 0–50% MeOH / DCM as the eluate. The pure fractions were combined and concentrated, and the resulting solid was co-evaporated with toluene and dried under high vacuum to obtain tris-triazole (4 g, 82%) as a white solid.

[0541] [ka] A stirred solution of alcohol 14 (0.68 g, 0.688 mmol) and diisopropylethylamine (0.51 g, 4.008 mmol) in DCM (10 mL) is mixed with 2-cyanoethyl=N,N-diisopropylchlorophosphoramidite (0.47 g, 2.004 mmol). mmol) was added dropwise. The reaction mixture was stirred at room temperature for 15 minutes. LC-MS indicated completion of the reaction. The reaction mixture was quenched with saturated NaHCO3 aqueous solution (20 mL), extracted with DCM (2 × 30 mL), washed with brine (20 mL), dried over Na2SO4, and evaporated. The crude product was then evaporated in a pre-equilibrated solution of (1% Et3N-MeOH:1% DCM (1:20) biotage silica gel. The product was loaded onto a ram (50 g 20 μm) and purified by flash chromatography using 0-30% MeOH / DCM with 1% Et3N as an additive. The pure fraction was collected and concentrated to obtain Amidite 15 (1.6 g 74%) as an off-white solid with >95% HPLC purity. The mass and NMR correspond to this product.

[0542] Example 18 [ka]

[0543] Example 19 [ka]

[0544] Example 20 [ka]

[0545] Example 21 Exemplary compounds falling within the scope of this disclosure could be synthesized according to the following scheme: [ka]

[0546] Example 22 [ka]

[0547] Example 23 [ka]

[0548] Exemplary compounds falling within the scope of this disclosure could be synthesized according to the following scheme: [ka]

[0549] References The contents of all references cited throughout this application (including references to

[0550] Equal portions Those skilled in the art will recognize, or confirm by conventional experimentation, many equivalents of the particular embodiments of the invention disclosed herein. Such equivalents are intended to be covered by the following claims.

[0551] Embodiment Further embodiments of the present invention include the following:

[0552] 1. Compounds of formula (Ia), (Ib), (Ic), or (Id), or their salts, solvates, or hydrates: [ka] And in equations (Ia), (Ib), (Ic), and (Id): [ka] is ring A, where each ring A is independently a substituted carbocyclyl or a substituted heterocyclyl as needed; [ka] is ring B, where each ring B is independently an optionally substituted aryl or optionally substituted heteroaryl; Each n is independently 0, 1, 2, 3, or 4; Each Y is independently O, CH2, S, S(=O), S(=O)2, NH, substituted amino, NHC(=O), C(=O)NH, P(=O)2-O-, P(=O)(=S)-O, P(=S)2-O, -OP(=O)2-O-, -OP(=O)(=S)-O-, -OP(=S)2-O-, -OP(=O)2-, -OP(=O)(=S)-, -OP(=S)2-; Each Z is independently an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, or ethylene glycol; Each R1 is independently an alkyl-O-phosphoramidite, an alkylphosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite; R2 and R5 are, independently, optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; R3 and R6 are each independently substituted with alkyl-O-GalNAc as needed. , optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; and Each R4 is independently alkyl-O-GalNAc as needed, alkenyl-O-GalNAc as needed, alkynyl-O-GalNAc as needed, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc. A compound, or its salt, solvate, or hydrate.

[0553] 2. The compound of Embodiment 1 having formula (Ia), or a salt, solvate, or hydrate thereof.

[0554] 3. The compound of Embodiment 1 or 2, wherein R2 and R3 are the same.

[0555] 4. The compound of Embodiment 1 having formula (Ic), or a salt, solvate, or hydrate thereof.

[0556] 5. Any compound from Embodiments 1 to 4, or a salt, solvate, or hydrate thereof, wherein the A ring is a substituted carbocykyl as needed.

[0557] 6. Any compound from Embodiments 1 to 4, or a salt, solvate, or hydrate thereof, wherein ring A is a heterocyclyl which may be substituted as needed.

[0558] 7. The compound of Embodiment 1 having formula (Ib), or a salt, solvate, or hydrate thereof.

[0559] 8. The compound of Embodiment 1 or 7, wherein R2 and R3 are the same.

[0560] 9. The compound of Embodiment 1 having formula (Id), or a salt, solvate, or hydrate thereof.

[0561] 10. A compound of any of Embodiments 1 or 7-9, or a salt, solvate, or hydrate thereof, wherein ring B is optionally substituted with an aryl compound.

[0562] 11. A compound of any of Embodiments 1 or 7-9, or a salt, solvate, or hydrate thereof, wherein ring B is a heteroaryl compound as needed.

[0563] 12. Compounds of formula (II-a), (II-b), or (II-c), or their salts, solvates, or hydrates: [ka] And in equations (II-a), (II-b), and (II-c): Each n is independently 0, 1, 2, 3, or 4; Each Y is independently O, CH2, S, S(=O), S(=O)2, NH, substituted N group, NHC(=O), C(=O)NH, P(=O)2-O-, P(=O)(=S)-O, P(=S)2-O, -OP(=O)2-O-, -OP(=O)(=S)-O-, -OP(=S)2-O-, -OP(=O)2-, -OP(=O)(=S)-, -OP(=S)2-; Each Z is independently an optionally substituted alkyl group, an optionally substituted alkenyl group, an optionally substituted alkynyl group, or ethylene glycol; Each R1 is independently an alkyl-O-phosphoramidite, an optionally substituted alkenylphosphoramidite, or an optionally substituted alkynylphosphoramidite; R5, R6, R7, and R8 are each independently of optionally substituted alkyl-O-GalNAc, polyethylene glycol-O-GalNAc, optionally substituted alkenyl-O-GalNAc, optionally substituted alkynyl-O-GalNAc, alkyl-S(=O)-alkyl-GalNAc, alkyl-S(=O)2-alkyl-GalNAc, alkyl-S(=O)-NH-alkyl, alkyl-S(=O)2-NH-alkyl-GalNAc, alkyl-PP(=O)(-O-)-NH-alkyl-GalNAc, alkyl-OP(=O)(-O-)-O-alkyl-GalNAc, alkyl-OP(-O-)(=S)-O-alkyl-GalNAc, or alkyl-OP(-S-)(=S)-O-alkyl-GalNAc; and R9 is H, adenine, guanine, thymine, cytosine, uracil, inosine (I), or a nucleic acid base variant. A compound, or its salt, solvate, or hydrate.

[0564] 13. The compound of Embodiment 12 having formula (II-a), or a salt, solvate, or hydrate thereof.

[0565] 14. The compound of Embodiment 12 having formula (II-b), or a salt, solvate, or hydrate thereof.

[0566] 15. The compound of Embo...

Claims

[Claim 1] The invention described in the specification.

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