Oligonucleotide composition and method thereof

Abstract

Above all, the present disclosure provides an oligonucleotide composition and a method thereof. In some embodiments, the oligonucleotide comprises various chemical modifications. In some embodiments, the present disclosure provides a technique useful for preparing the oligonucleotide.

Description

Background Technology

[0001] Cross-reference regarding related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 620,768 filed January 12, 2024, the entirety of which is incorporated herein by reference.

[0003] Oligonucleotides are useful for many purposes. Various techniques for manufacturing oligonucleotides have been reported.

[0004] Various chemical modifications have been reported to enhance the properties and activity of oligonucleotides. For example, -P(O)(O - A 5'-terminal -P(O)(OH)2 group, which may exist in a salt form such as )2, has been incorporated into the 5'-terminus of various oligonucleotides in some cases along with another chemical moiety to enhance the properties and / or activity of the oligonucleotides, such as RNAi agents. In some embodiments, the oligonucleotide comprises the following group attached to a carbon atom at the 5'-terminus, wherein the phosphonate is bonded to a triazole ring:

[0005] .

[0006] In some embodiments, it exists in the form of a salt as an anion:

[0007] .

[0008] Various techniques for incorporating phosphonate groups have been reported. Typically, during oligonucleotide synthesis, -P(O)(OH)2 groups are incorporated into protected groups (e.g., -P(O)(OMe)2, -P(O)(OEt)2, etc.) that are later deprotected. Among other things, the present disclosure includes the recognition that it may be difficult to deprotect such protected groups under typical oligonucleotide synthesis conditions. In some embodiments, additional steps, harsher conditions, and / or longer reaction times are required. In some embodiments, such protected groups result in lower oligonucleotide yield and / or purity. In some embodiments, such protected phosphate groups increase operational complexity. In some embodiments, such protected phosphate groups reduce manufacturing efficiency. In some embodiments, such protected phosphate groups increase manufacturing costs. In some embodiments, the present disclosure is provided to address these challenges.

[0009] In some embodiments, the present disclosure provides a technique for incorporating -P(O)(OH)2 into oligonucleotides with higher yield, higher purity, higher efficiency, lower operational complexity, milder reaction conditions, shorter manufacturing time and / or lower cost, etc., compared to using, for example, -P(O)(OMe)2 or -P(O)(OEt)2. In some embodiments, the present disclosure provides a protected -P(O)(OH)2 group that can be deprotected under oligonucleotide synthesis conditions during cleavage and / or deprotection, which is utilized, for example, for oligonucleotides lacking the -P(O)(OH)2 group.

[0010] In some embodiments, the present disclosure is -P(O)(OR PG It provides a technology including )2, wherein each R PGis a group that is unstable under oligonucleotide synthesis conditions during cleavage and / or deprotection, utilized, for example, for oligonucleotides lacking a -P(O)(OH)2 group. In some embodiments, each R PG is a group that can be removed during cleavage and / or deprotection, utilized under oligonucleotide synthesis conditions, for example, for oligonucleotides lacking the -P(O)(OH)2 group. In some embodiments, -P(O)(OR PG )2 is a protected -P(O)(OH)2 group that is converted to -P(O)(OH)2, which may exist in salt form during cleavage and / or deprotection, utilized, for example, for oligonucleotide-free oligonucleotides under oligonucleotide synthesis conditions. In some embodiments, the P atom is bonded to a carbon atom.

[0011] For example, in some implementations, R PG is 1) an alpha carbon atom bonded to the oxygen to which it is attached (where the alpha carbon atom is bonded to hydrogen), and 2) a beta carbon atom (where the beta carbon atom is an electron-withdrawing group like R (e.g., compared to ethyl). PG R that can facilitate the removal of PG11 Includes (coupled to the device). In some embodiments, R PG11 is an electronic absorber. In some embodiments, R PG11 R as described herein s is. In some implementations, R PG11 is -CN. In some implementations, R PG11 -L s -R s11 and, here, each L s and R s11 ...is as independently described herein. In some embodiments, R PG is -CH2-R s and, where -CH2- is independently and optionally substituted, and R sis as described herein. In some embodiments, -CH2- is optionally a single substitution. In some embodiments, R PG R as described herein P1 is. In some implementations, R PG R as described herein P2 is. In some implementations, R PG is -O-CH2CH2CN.

[0012] In some implementations, R PG is a labile group. In some embodiments, R PG -CH2-O(CO)R PG12 and, here R PG12 is a trisubstituted methyl and -CH2- is optionally substituted. In some embodiments, R PG -CH2-O(CO)R PG12 and, here R PG12 is -C(R s11 )3 and here, each R s11 ...is as independently described herein. In some embodiments, R PG is R PG12 and, here R PG12 is a trisubstituted methyl. In some embodiments, R PG12 is -C(R s11 )3 and here, each R s11 ...is as independently described herein. In some embodiments, trisubstituted methyl or -C(R s11 The carbon of )3 is bonded to three carbon atoms. In some embodiments, R PG12 is an optionally substituted t-butyl. In some embodiments, R PG12 is t-butyl. In some embodiments, R PG R as described herein P1 or R P2 is. In some implementations, R PG is -O-CH2-OC(O)-R P and, here R Pis an optionally substituted tert-butyl. In some embodiments, R PG is -O-CH2-OC(O)-tBu.

[0013] In some implementations, -P(O)(OR PG )2 is -P(O)(R as described herein (e.g., in Formula I or Formula I'). P1 )(R P2 It has the structure of ), where R 1 and R P2 Each is as described independently in the present invention.

[0014] In some embodiments, the present disclosure is as -P(O)(OR PG An oligonucleotide comprising -P(O)(OH)2 is provided. In some embodiments, such oligonucleotides are useful for preparing or delivering oligonucleotides comprising -P(O)(OH)2 which may exist in various salt forms. In some embodiments, -P(O)(OR as described herein PG Oligonucleotides containing )2 are administered to a subject to prevent or treat a pathological condition, disorder, or disease. In some embodiments, -P(O)(OR as described herein PG Oligonucleotides containing -P(O)(OH)2 are utilized to prepare oligonucleotides containing -P(O)(OH)2, which may exist in various salt forms. For example, in some embodiments, the present disclosure is Provides a compound that is an oligonucleotide containing, wherein R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is optionally substituted tert-butyl. In some embodiments, the present disclosure provides a compound that is an oligonucleotide comprising a moiety of formula OI or a salt thereof:

[0015] ,

[0016] OI

[0017] Here:

[0018] R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl;

[0019] BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase;

[0020] R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a , -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s is;

[0021] Each L 2s is independently L and;

[0022] Each R 2a is independently R' and;

[0023] Each L is independently C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-;

[0024] -Cy- is an optionally substituted divalent 3-20 member ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur;

[0025] Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' are taken together with an intervening atom(s) to form an optionally substituted 3- to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the intervening atom(s); and

[0026] Each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 It is an optionally substituted group selected from a 5-14 member heteroaryl ring having 1-5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3-15 member heterocyclic ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur.

[0027] In some embodiments, the present disclosure is -P(O)(OR PG Provides a nucleoside phosphoramidite containing )2. In some embodiments, -P(O)(OR PG)2 is bonded to the sugar of the nucleoside (e.g., the 5' carbon). In some embodiments, it is bonded to another moiety bonded to the sugar of the nucleoside (e.g., the 5' carbon), e.g., an optionally substituted triazole ring (e.g., as in Formula I or Formula I'). In some embodiments, the phosphoramidite comprises a chiral auxiliary.

[0028] In some embodiments, the provided technology provides a compound of formula I' or a salt thereof:

[0029] ,

[0030] I'

[0031] Here:

[0032] R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl;

[0033] SU is a party (e.g., , here R 2s is as described herein) and;

[0034] BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase;

[0035] R 1 , R 2 and R 3 Each is independently R' or R 1 , R 2 and R 3 Two or three of them are taken together with the atoms interposed between them Forming;

[0036] Ring A is an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms;

[0037] t is 0~5 and;

[0038] Each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R s11 , -L s -OR s11 , -L s -SR s11 , -L s -N(R s11 )2, -OL s -OR s11 , -OL s -SR s11 , -OL s -N(R s11 )2, -C(R s11 )3 or -L s -Si(R s11 )3 and;

[0039] Each R s11 is independently R' and;

[0040] Each L s is independently L and;

[0041] Each L is independently covalently bonded, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-;

[0042] -Cy- is an optionally substituted divalent 3-20 member ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur;

[0043] Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' are taken together with an intervening atom(s) to form an optionally substituted 3- to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the intervening atom(s); and

[0044] Each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 It is an optionally substituted group selected from a 5-14 member heteroaryl ring having 1-5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3-15 member heterocyclic ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur.

[0045] In some embodiments, the provided technology provides a compound of formula I or a salt thereof:

[0046] ,

[0047] I

[0048] Here:

[0049] R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl;

[0050] BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase;

[0051] R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a , -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s is;

[0052] Each L 2s is independently L and;

[0053] Each R 2a is independently R' and;

[0054] Each L is independently covalently bonded, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-;

[0055] -Cy- is an optionally substituted divalent 3-20 member ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur;

[0056] R 1 , R 2 and R 3 Each is independently R' or R 1 , R 2 and R 3 Two or three of them are taken together with the atoms interposed between them Forming;

[0057] Ring A is an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms;

[0058] t is 0~5 and;

[0059] Each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R s11 , -L s -OR s11 , -L s -SR s11 , -L s -N(R s11 )2, -OL s -OR s11 , -OL s -SR s11 , -OL s -N(R s11 )2, -C(R s11 )3 or -L s -Si(R s11 )3 and;

[0060] Each R s11 is independently R' and;

[0061] Each L s is independently L and;

[0062] Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' are taken together with an intervening atom(s) to form an optionally substituted 3- to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the intervening atom(s); and

[0063] Each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 It is an optionally substituted group selected from a 5-14 member heteroaryl ring having 1-5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3-15 member heterocyclic ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur.

[0064] In some embodiments, the compound of Formula I or a salt thereof is useful as a nucleoside phosphoramidite for oligonucleotide synthesis. In some embodiments, the compound of Formula I' or a salt thereof is useful as a nucleoside phosphoramidite for oligonucleotide synthesis.

[0065] In some embodiments, the present disclosure is -P(O)(OR PG Provides a nucleoside containing )2. In some embodiments, -P(O)(OR PG )2 is bonded to the sugar of the nucleoside (e.g., the 5' carbon). In some embodiments, it is bonded to another moiety bonded to the sugar of the nucleoside (e.g., the 5' carbon), e.g., to an optionally substituted triazole ring (e.g., as in Formula I or Formula I'). In some embodiments, the present disclosure provides a compound of Formula II' or a salt thereof:

[0066] ,

[0067] II'

[0068] Here:

[0069] R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl;

[0070] SU is a party (e.g., , here R 2s is as described herein) and; and

[0071] BA is an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase.

[0072] In some embodiments, the present disclosure provides a compound of Formula II or a salt thereof:

[0073] ,

[0074] II

[0075] Here:

[0076] R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl;

[0077] BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase;

[0078] R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a, -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s is;

[0079] Each L 2s is independently L and;

[0080] Each R 2a is independently R' and;

[0081] Each L is independently C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-;

[0082] -Cy- is an optionally substituted divalent 3-20 member ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur;

[0083] Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' are taken together with an intervening atom(s) to form an optionally substituted 3- to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the intervening atom(s); and

[0084] Each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 It is an optionally substituted group selected from a 5- to 14-membered heteroaryl ring having 1- to 5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3- to 15-membered heterocyclic ring having 1- to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, the present disclosure provides techniques for preparing oligonucleotides and compositions thereof. In some embodiments, the oligonucleotide comprises -P(O)(OH)2 which may exist in various salt forms. In some embodiments, the oligonucleotide comprises -P(O)(OR as described herein PG Includes )2. In some embodiments, the present disclosure provides a technique for manufacturing phosphoramidite. In some embodiments, the present disclosure provides a technique for manufacturing nucleosides. Specific details for implementing the invention

[0085] The technology of the present disclosure can be more easily understood by referring to the following detailed description of specific embodiments.

[0086] definition

[0087] As used herein, the following definitions will apply unless otherwise indicated. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. Additionally, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999 and "March's Advanced Organic Chemistry", 5th edition, Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001.

[0088] As used in this disclosure, unless otherwise evident from the context, (i) the terms “a” or “an” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and / or”; (iii) the terms “comprising,” “comprising,” “comprising” (whether or not used with “not limited thereto”) and “comprising” (whether or not used with “not limited thereto”) may be understood to include an itemized component or step presented either by itself or together with one or more additional components or steps; (iv) the term “another” may be understood to mean at least one additional / second one or more; (v) the terms “about” and “approximately” may be understood to allow for standard variations understood by those skilled in the art; and (vi) both end values ​​are included where a range is provided.

[0089] Unless otherwise specified, the description of oligonucleotides and their elements (e.g., base sequence, sugar modification, internucleotide linkage, linkage stereochemistry, pattern thereof, etc.) is from 5' to 3'. As understood by those skilled in the art, in some embodiments, oligonucleotides may be provided and / or used in salt form, particularly in pharmaceutically acceptable salt forms, e.g., as sodium salts. As also recognized by those skilled in the art, in some embodiments, individual oligonucleotides in a composition may be considered to have the same composition and / or structure, even though specific such oligonucleotides may exist in different salt form(s) at a given moment within such composition (e.g., liquid composition) (and may be soluble and the oligonucleotide chains may exist in anionic form (e.g., when in a liquid composition)). For example, those skilled in the art will recognize that at a given pH, the linkages between individual nucleotides along an oligonucleotide chain may exist in the acid (H) form, or in a plurality of possible salt forms (e.g., sodium salts, or salts of different cations (depending on which ions may be present in the formulation or composition)), and that the acid form thereof (e.g., all cations (if any) are H + If (replaced by) is of the same composition and / or structure, these individual oligonucleotides can appropriately be considered to be of the same composition and / or structure.

[0090] Aliphatic: As used herein, “aliphatic” means a straight (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units (but is not aromatic), or a substituted or unsubstituted monocyclic, dicyclic, or polycyclic hydrocarbon ring that is fully saturated or contains one or more unsaturated units (but is not aromatic), or a combination thereof. In some embodiments, the aliphatic group comprises 1 to 50 aliphatic carbon atoms. In some embodiments, the aliphatic group comprises 1 to 20 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 10 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 9 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 8 aliphatic carbon atoms. In other embodiments, the aliphatic group comprises 1 to 7 aliphatic carbon atoms. In another embodiment, the aliphatic group comprises 1 to 6 aliphatic carbon atoms. In yet another embodiment, the aliphatic group comprises 1 to 5 aliphatic carbon atoms, and in yet another embodiment, the aliphatic group comprises 1, 2, 3, or 4 aliphatic carbon atoms. Suitable aliphatic groups include linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof, e.g., (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0091] Alkenyl: As used herein, the term “alkenyl” refers to an aliphatic group having one or more double bonds as defined herein.

[0092] Alkyl: As used herein, the term "alkyl" is given its ordinary meaning in the art and may include straight-chain alkyl groups, branched-chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, and saturated aliphatic groups including cycloalkyl-substituted alkyl groups. In some embodiments, the alkyl has 1 to 100 carbon atoms. In specific embodiments, the straight-chain or branched-chain alkyl has about 1 to 20 carbon atoms in its backbone (e.g., C1-C in the case of a straight chain). 20 , in the case of branched chains, C2-C 20 ), and alternatively, have about 1 to 10 carbon atoms. In some embodiments, the cycloalkyl ring has about 3 to 10 carbon atoms in its ring structure, and such ring is monocyclic, bicyclic, or polycyclic, and alternatively, the ring structure has about 5, 6, or 7 carbons. In some embodiments, the alkyl group may be a lower alkyl group, wherein the lower alkyl group comprises 1 to 4 carbon atoms (e.g., C1-C4 in the case of a straight-chain lower alkyl).

[0093] Alkynyl: As used herein, the term “alkynyl” refers to an aliphatic group having one or more triple bonds as defined herein.

[0094] Aryl: The term “aryl” as used herein, used alone or as part of a macromoiety as in “araclel,” “aralkoxy,” or “aryloxyalkyl,” refers to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, wherein at least one ring in the system is aromatic. In some embodiments, the aryl group is a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 14 ring members, wherein at least one ring in the system is aromatic and each ring in the system comprises 3 to 7 ring members. In some embodiments, each monocyclic ring unit is aromatic. In some embodiments, the aryl group is a biaryl group. The term “aryl” may be used interchangeably with the term “aryl ring.” In specific embodiments of the present disclosure, “aryl” refers to an aromatic ring system including, but not limited to, phenyl, biphenyl, naphthyl, binaphthyl, anthracyl, etc., which may have one or more substituents. Additionally, as used herein, the scope of the term “aryl” includes a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, etc.

[0095] Chiral Control: As used herein, "chiral control" refers to the control of the stereochemical designation of a chiral linker in a chiral nucleotide-to-chiral linkage within an oligonucleotide. As used herein, a chiral nucleotide-to-chiral linkage is a linkage between nucleotides in which the linker is chiral. In some embodiments, control is achieved through a chiral element absent in the sugar and base moiety of the oligonucleotide, for example, in some embodiments, control is achieved through the use of one or more chiral aids during the preparation of the oligonucleotide, said chiral aids are often parts of a chiral phosphoramidite used during the preparation of the oligonucleotide. In contrast to chiral control, those skilled in the art will understand that if conventional oligonucleotide synthesis without the use of chiral aids is used to form chiral nucleotide-to-chiral linkages, such conventional oligonucleotide synthesis cannot control the stereochemistry in the chiral nucleotide-to-chiral linkages. In some embodiments, the stereochemical designation of each chiral linker in each chiral nucleotide-to-chiral linkage within the oligonucleotide is controlled.

[0096] Chiral-controlled oligonucleotide composition: As used herein, the terms “chiral-controlled oligonucleotide composition,” “chiral-controlled nucleic acid composition,” and similar terms mean a composition comprising a plurality of oligonucleotides (or nucleic acids) sharing a common base sequence, wherein the plurality of oligonucleotides (or nucleic acids) share the same linkage stereochemistry in one or more chiral nucleotide-links (chiral-controlled or stereodefined nucleotide-links, the chiral linkage factor is Rp or Sp in the composition (“stereographically defined”), and is not a random mixture of Rp and Sp such as non-chiral-controlled nucleotide-links). In some embodiments, the chiral-controlled oligonucleotide composition comprises a plurality of oligonucleotides (or nucleic acids) sharing the following: 1) a common base sequence, 2) a common pattern of backbone linkage, and 3) a common pattern of backbone phosphate modification, wherein the plurality of oligonucleotides (or nucleic acids) share the same linkage phosphate stereochemistry in one or more chiral nucleotide-linkages (chiral-controlled or stereodefined nucleotide-linkages, the chiral linkage phosphate is Rp or Sp in the composition ("stereographically defined"), and is not a random mixture of Rp and Sp such as in non-chiral-controlled nucleotide-linkages). The levels of the plurality of oligonucleotides (or nucleic acids) in the chiral-controlled oligonucleotide composition are predetermined / controlled or abundant compared to random levels in the chiral-uncontrolled oligonucleotide composition (e.g., through the preparation of chiral-controlled oligonucleotides to stereoselectively form one or more chiral nucleotide-linkages).In some embodiments, about 1% to 100% of all oligonucleotides in the chiral-controlled oligonucleotide composition (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) are multiple oligonucleotides. In some embodiments, about 1% to 100% of all oligonucleotides in a chiral-controlled oligonucleotide composition sharing a common base sequence, a common pattern of backbone linkage, and a common pattern of backbone phosphorus modification (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) are multiple oligonucleotides.In some embodiments, the level is about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 10%, 50% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5% to 90% of all oligonucleotides in the composition sharing a common base sequence (e.g., a plurality of oligonucleotides or oligonucleotide types), or all oligonucleotides in the composition sharing a common base sequence, a common pattern of backbone linkage, and a common pattern of backbone modification, or all oligonucleotides in the composition sharing a common base sequence, a common base modification pattern, a common sugar modification pattern, a common internucleotide linkage type pattern, and / or a common internucleotide linkage modification pattern) of all oligonucleotides in the composition sharing a common base sequence, a common base modification pattern, a common sugar modification pattern, a common internucleotide linkage type pattern, and / or a common internucleotide linkage modification pattern. It is about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%). In some embodiments, a plurality of oligonucleotides share the same stereochemistry in the linkage between about 1 to 50 (e.g., about 1 to 10, 1 to 20, 5 to 10, 5 to 20, 10 to 15, 10 to 20, 10 to 25, 10 to 30, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, or at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) chiral nucleotides.In some embodiments, a plurality of oligonucleotides are about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, or at least They share the same stereochemistry in the linkages between chiral nucleotides of 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%). In some embodiments, where applicable, multiple oligonucleotides (or nucleic acids) share the same pattern of sugar and / or nucleobase modifications. In some embodiments, multiple oligonucleotides (or nucleic acids) are various forms of the same oligonucleotide (e.g., acids and / or various salts of the same oligonucleotide). In some embodiments, multiple oligonucleotides (or nucleic acids) have the same composition.In some embodiments, the level of a plurality of oligonucleotides (or nucleic acids) is about 1% to 100% (e.g., about 5% to 100%, 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 95% to 100%, 50% to 90%, or about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or at least 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%). In some embodiments, the linkage between each chiral nucleotide is a chiral-controlled linkage between nucleotides, and the composition is a completely chiral-controlled oligonucleotide composition. In some embodiments, a plurality of oligonucleotides (or nucleic acids) are structurally identical. In some embodiments, the chiral-controlled internucleotide linkage has a diastereometric purity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%, and generally at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereometric purity of at least 95%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereometric purity of at least 96%. In some embodiments, the chiral-controlled internucleotide linkage has a diastereometric purity of at least 97%. In some embodiments, the chiral-controlled internucleotide linkage has at least 98% distereometric purity.In some embodiments, the chiral-controlled internucleotide linkage has at least 99% diastereform purity. In some embodiments, the percentage of the level is (DS). nc That is all, DS is a partial stereopurity as described in the present invention (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or more), and nc is the number of chiral linkages as described in the present invention (e.g., 1–50, 1–40, 1–30, 1–25, 1–20, 5–50, 5–40, 5–30, 5–25, 5–20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, It is 25 or higher). In some implementations, the percentage of the level is (DS) nc That is all, DS is a partial stereopurity as described in the present invention (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% or more), and nc is the number of chiral-controlled nucleotide linkages as described in the present invention (e.g., 1–50, 1–40, 1–30, 1–25, 1–20, 5–50, 5–40, 5–30, 5–25, 5–20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, It is 24, 25 or higher). In some implementations, the percentage of the level is (DS) nc It is greater than or equal to, and DS is 95%–100%. For example, if DS is 99% and nc is 10, the percentage is 90% or greater ((99%) 10 0.90 = 90%). In some embodiments, the levels of multiple oligonucleotides in the composition are expressed as the product of the diastereferences of each chiral linkage. In some embodiments, the levels of multiple oligonucleotides in the composition are expressed as the product of the diastereferences of each chiral-controlled internucleotide linkage in the oligonucleotides. In some embodiments, the diastereference of the internucleotide linkage connecting two nucleosides in the oligonucleotide (or nucleic acid) is expressed as the diastereference of the internucleotide linkage of the dimer connecting the same two nucleosides, and such dimers are prepared using comparable conditions, in some cases the same synthesis cycle conditions (e.g., in the case of the linkage between Nx and Ny in oligonucleotide …NxNy…, the dimer is NxNy). In some embodiments, not all chiral internucleotide linkages are chiral-controlled internucleotide linkages, and the composition is a diastereference-controlled oligonucleotide composition. In some embodiments, non-chiral controlled internucleotide linkages have a partial stereopurity of about 80%, 75%, 70%, 65%, 60%, less than 55%, or about 50%, as is generally observed in stereorandom oligonucleotide compositions (e.g., from conventional oligonucleotide synthesis, e.g., the phosphoramidite method, as understood by those skilled in the art). In some embodiments, multiple oligonucleotides (or nucleic acids) are of the same type. In some embodiments, the chiral controlled oligonucleotide composition comprises individual oligonucleotide or nucleic acid types at nonrandom or controlled levels. For example, in some embodiments, the chiral controlled oligonucleotide composition comprises one or fewer oligonucleotide types. In some embodiments, the chiral controlled oligonucleotide composition comprises more than one oligonucleotide type. In some embodiments, the chiral controlled oligonucleotide composition comprises multiple oligonucleotide types.In some embodiments, the chiral-controlled oligonucleotide composition is a composition of oligonucleotides of a predetermined oligonucleotide type, and the composition comprises a plurality of oligonucleotides of the said oligonucleotide type at non-random or controlled levels.

[0097] Comparable: As applied herein, the term “comparable” is used to describe two sets (or more) of conditions or situations that are sufficiently similar to one another to enable a comparison of the obtained results or observed phenomena. In some embodiments, the sets of comparable conditions or situations feature a plurality of substantially identical features and one or a few altered features. Those skilled in the art will understand that sets of conditions are similar to one another if they feature a number and type of substantially identical features sufficient to warrant a reasonable conclusion that the difference in the obtained results or observed phenomena under different sets of conditions or situations is caused by or indicates a variation in the altered features.

[0098] Cyclic: The terms “cyclic,” “carbocycle,” “carbocyclile,” “carbon-cyclic radical,” and “carbon-cyclic ring” are used interchangeably and, as used herein, unless otherwise specified, refer to a saturated or partially unsaturated but non-aromatic cyclic aliphatic monocyclic, dicyclic, or polycyclic ring system having 3 to 30 ring members as described herein. The cyclocyclic group comprises, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, adamantyl, and cyclooctadienyl. In some embodiments, the cyclocyclic group has 3 to 6 carbons. In some embodiments, the cyclocyclic group is saturated and is cycloalkyl. Additionally, the term “aliphatic” may include an aliphatic ring fused to one or more aromatic or non-aromatic rings, such as decahydronaphthyl or tetrahydronaphthyl. In some embodiments, the aliphatic group is dicyclic. In some embodiments, the aliphatic group is tricyclic. In some embodiments, the aliphatic group is polycyclic. In some embodiments, the “aliphatic” is a C3-C6 monocyclic hydrocarbon, or a C8-C6 hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single attachment site to the rest of the molecule. 10 A cyclic or polycyclic hydrocarbon, or a C9-C that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single attachment site to the rest of the molecule 16 It refers to polycyclic hydrocarbons.

[0099] Heteroaliphatic: As used herein, the term “heteroaliphatic” is given its ordinary meaning in the art and refers to an aliphatic group as described herein in which one or more carbon atoms are independently replaced by one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). In some embodiments, one or more units selected from C, CH, CH2, and CH3 are independently replaced by one or more heteroatoms (including their oxidized and / or substituted forms). In some embodiments, the heteroaliphatic group is heteroalkyl. In some embodiments, the heteroaliphatic group is heteroalkenyl.

[0100] Heteroalkyl: As used herein, the term “heteroalkyl” is given its ordinary meaning in the art and refers to an alkyl group as described herein in which one or more carbon atoms are independently replaced by one or more heteroatoms (e.g., oxygen, nitrogen, sulfur, silicon, phosphorus, etc.). Examples of heteroalkyl groups include, but are not limited to, alkoxy, poly(ethylene glycol)-, alkyl-substituted amino, tetrahydrofuranyl, piperidinyl, morpholinyl, etc.

[0101] Heteroaryl: As used herein, the terms “heteroaryl” and “heteroar-”, used alone or as part of a larger moiety (e.g., “heteroaralkyl” or “heteroaralkoxy”), refer to a monocyclic, bicyclic, or polycyclic ring system having a total of 5 to 30 ring members, wherein at least one ring in the system is aromatic and at least one aromatic ring atom is a heteroatom. In some embodiments, the heteroaryl group is a group having 5 to 10 ring atoms (i.e., monocyclic, bicyclic, or polycyclic), and in some embodiments, 5, 6, 9, or 10 ring atoms. In some embodiments, each monocyclic ring unit is aromatic. In some embodiments, the heteroaryl group has 6, 10, or 14 π electrons shared in a cyclic arrangement; and has 1 to 5 heteroatoms in addition to carbon atoms. The heteroaryl group comprises, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, furinyl, naphthiridinyl, and pteridinyl. In some embodiments, the heteroaryl is a heterobiaryl group, e.g., bipyridyl. As used herein, the terms “heteroaryl” and “heteroar-” also comprise a group in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, wherein the radical or attachment site is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolidinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazine-3(4H)-one. The heteroaryl group may be monocyclic, bicyclic, or polycyclic.The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which includes an optionally substituted ring. The term "heteroaralkyl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions are independently and optionally substituted.

[0102] Heteroatom: As used herein, the term "heteroatom" means an atom that is not carbon or hydrogen. In some embodiments, the heteroatom is boron, oxygen, sulfur, nitrogen, phosphorus, or silicon (including oxidized forms of nitrogen, sulfur, phosphorus, or silicon; charged forms of nitrogen (e.g., quaternized forms, forms such as imium groups, etc.), phosphorus, sulfur, oxygen; etc.). In some embodiments, the heteroatom is silicon, phosphorus, oxygen, sulfur, or nitrogen. In some embodiments, the heteroatom is silicon, oxygen, sulfur, or nitrogen. In some embodiments, the heteroatom is oxygen, sulfur, or nitrogen.

[0103] Heterocyclic: As used herein, the terms “heterocyclic,” “heterocyclic radical,” and “heterocyclic ring” (as used herein) are used interchangeably and refer to a monocyclic, dicyclic, or polycyclic ring moiety (e.g., 3 to 30 members) that is saturated or partially unsaturated and has one or more heterocyclic ring atoms. In some embodiments, the heterocyclic group is a stable 5 to 7-membered monocyclic or 7 to 10-membered dicyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more, preferably 1 to 4, heterocyclic atoms in addition to the carbon atom, as defined above. When used in relation to the ring atoms of the heterocyclic, the term “nitrogen” includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, and nitrogen, nitrogen is N (as in 3,4-dihydro-2H-pyrrolil), NH (as in pyrrolidinyl), or+ It may be NR (as in N-substituted pyrrolidinyl). The heterocyclic ring may be attached to its pendant group at any heteroatom or carbon atom that forms a stable structure, and any of the ring atoms may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazefinyl, oxazefinyl, thiazefinyl, morpholinyl, and quinuclidinyl. The terms “heterocyclic,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclic group,” “heterocyclic moiety,” and “heterocyclic radical” are used interchangeably herein and also comprise a heterocyclyl ring fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl. The heterocyclyl group may be monocyclic, bicyclic, or polycyclic. The term “heterocyclylalkyl” refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions are independently and optionally substituted.

[0104] Internucleotide linkage: As used herein, the phrase “internucleotide linkage” generally refers to a linkage connecting nucleoside units of an oligonucleotide or nucleic acid. In some embodiments, the internucleotide linkage is a phosphodiester linkage, as is widely found in naturally occurring DNA and RNA molecules (a natural phosphate linkage (-OP(=O)(OH)O-)) which may exist in salt form as understood by those skilled in the art). In some embodiments, the internucleotide linkage is a modified internucleotide linkage (not a natural phosphate linkage). In some embodiments, the internucleotide linkage is a “modified internucleotide linkage” in which at least one oxygen atom or -OH of the phosphodiester linkage is replaced by another organic or inorganic moiety. In some embodiments, such organic or inorganic moiety is =S, =Se, =NR', -SR', -SeR', -N(R')2, B(R') 3,-S-, -Se-, and -N(R')-, wherein each R' is independently defined and described in this disclosure. In some embodiments, the nucleotide linkage is a phosphotriester linkage, a phosphorothioate linkage (or a phosphorothioate diester linkage, -OP(=O)(SH)O-, which may exist in salt form as recognized by those skilled in the art), or a phosphorothioate tryster linkage. In some embodiments, the modified nucleotide linkage is a phosphorothioate linkage. In some embodiments, the nucleotide linkage is, for example, one of a PNA (peptide nucleic acid) or PMO (phosphorodiamidate morpholino oligomer) linkage. In some embodiments, the modified nucleotide linkage is a non-negatively charged nucleotide linkage. In some embodiments, the modified internucleotide linkage is a neutral internucleotide linkage (e.g., n001 in the specific oligonucleotides provided). Those skilled in the art understand that the internucleotide linkage may exist as an anion or a cation at a given pH due to the presence of an acid or base moiety in the linkage.

[0105] Linking Phosphor: As defined herein, the phrase “linking phosphate” is used to indicate that the specific phosphate atom referred to is a phosphate atom present in a nucleotide-nucleotide linkage, and this phosphate atom corresponds to the phosphate atom in phosphodiester-nucleotide linkages occurring in naturally occurring DNA and RNA. In some embodiments, the linking phosphate atom is present in a modified nucleotide-nucleotide linkage, wherein each oxygen atom of the phosphodiester linkage is optionally and independently replaced by an organic or inorganic moiety. In some embodiments, the linking phosphate atom is chiral (e.g., as in phosphorothioate-nucleotide linkages). In some embodiments, the linking phosphate atom is achiral (e.g., as in natural phosphate linkages).

[0106] Modified nucleobase: The terms "modified nucleobase," "modified base," etc. refer to a chemical moiety that is chemically distinct from a nucleobase but can perform at least one function of a nucleobase. In some embodiments, the modified nucleobase is a nucleobase that includes a modification. In some embodiments, the modified nucleobase is capable of at least one function of a nucleobase and can form a moiety in a polymer capable of forming base pairs with, for example, a nucleic acid containing at least a complementary base sequence. In some embodiments, the modified nucleobase is a substituted A, T, C, G, or U, or a substituted tautomer of A, T, C, G, or U. In some embodiments, in the context of an oligonucleotide, the modified nucleobase refers to a nucleobase that is not A, T, C, G, or U.

[0107] Modified nucleoside: The term “modified nucleoside” refers to a moiety that is derived from or chemically similar to a natural nucleoside but includes chemical modifications that distinguish it from a natural nucleoside. Non-limiting examples of modified nucleosides include modifications in bases and / or sugars. Non-limiting examples of modified nucleosides include having a 2’ modification in a sugar. Non-limiting examples of modified nucleosides also include baseless nucleosides (lacking a nucleobase). In some embodiments, the modified nucleoside may perform at least one function of a nucleoside and may form a moiety in a polymer capable of forming base pairs with a nucleic acid containing, for example, at least a complementary base sequence.

[0108] Modified nucleotide: The term “modified nucleotide” comprises any chemical moiety that is structurally different from a natural nucleotide but can perform at least one function of a natural nucleotide. In some embodiments, the modified nucleotide comprises modifications in the sugar, base, and / or nucleotide linkage. In some embodiments, the modified nucleotide comprises a modified sugar, a modified nucleobase, and / or a modified nucleotide linkage. In some embodiments, the modified nucleotide can perform at least one function of a nucleotide and can form a subunit in a polymer capable of forming base pairs with a nucleic acid comprising, for example, at least a complementary base sequence.

[0109] Modified Sugar: The term "modified sugar" refers to a moiety that can replace a sugar. A modified sugar mimics the spatial arrangement, electronic properties, or some other physicochemical properties of a sugar. In some embodiments, as described in this disclosure, the modified sugar is substituted ribose or deoxyribose. In some embodiments, the modified sugar includes a 2'-modification. Examples of useful 2'-modifications are widely used in the art and are described herein. In some embodiments, the 2'-modification is 2'-F. In some embodiments, the 2'-modification is 2'-OR, where R is optionally substituted C 1-10 It is aliphatic. In some embodiments, the 2'-modification is 2'-OMe. In some embodiments, the 2'-modification is 2'-MOE. In some embodiments, the modified sugar is a dicyclic sugar (e.g., a sugar used in LNA, BNA, etc.). In some embodiments, with respect to the oligonucleotide, the modified sugar is a sugar other than ribose or deoxyribose, which are commonly found in natural RNA or DNA.

[0110] Nucleic Acid: As used herein, the term “nucleic acid” includes any nucleotide and polymers thereof. As used herein, the term “polynucleotide” refers to a polymeric form of nucleotides of any length, such as ribonucleotides (RNA) or deoxyribonucleotides (DNA) or combinations thereof. These terms refer to the primary structure of molecules and thus include double-stranded and single-stranded DNA, and double-stranded and single-stranded RNA. These terms include, as equivalents, analogs of RNA or DNA comprising modified nucleotides and / or modified polynucleotides, such as but not limited to methylated, protected, and / or capped nucleotides or polynucleotides. The terms include poly- or oligo-ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from N-glycosides or C-glycosides of nucleobases and / or modified nucleobases; nucleic acids derived from sugars and / or modified sugars; and includes nucleic acids derived from phosphate crosslinks and / or modified internucleotide linkages. The term includes nucleic acids containing any combination of a nucleobase, a modified nucleobase, a sugar, a modified sugar, a phosphate crosslink, or a modified internucleotide linkage. Examples include, but are not limited to, nucleic acids containing a ribose moiety, nucleic acids containing a deoxyribose moiety, nucleic acids containing both ribose and deoxyribose moiety, and nucleic acids containing ribose and a modified ribose moiety. Unless otherwise specified, the prefix poly- refers to nucleic acids containing 2 to about 10,000 nucleotide monomer units, and the prefix oligo- refers to nucleic acids containing 2 to about 200 nucleotide monomer units.

[0111] Nucleobase: The term "nucleobase" refers to a portion of nucleic acid involved in hydrogen bonding that binds one nucleic acid strand to another complementary strand in a sequence-specific manner. The most common naturally occurring nucleosides are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, the naturally occurring nucleobase is a modified adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the naturally occurring nucleobase is methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the nucleobase comprises a heteroaryl ring in which the ring atom is nitrogen, and when in a nucleoside, the nitrogen is bound to the sugar moiety. In some embodiments, the nucleobase comprises a heterocyclic ring in which the ring atom is nitrogen, and when in a nucleoside, the nitrogen is bound to the sugar moiety. In some embodiments, the nucleobase is a “modified nucleobase” that is a nucleobase other than adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T). In some embodiments, the modified nucleobase is a substituted A, T, C, G, or U. In some embodiments, the modified nucleobase is a substituted tautomer of A, T, C, G, or U. In some embodiments, the modified nucleobase is a methylated adenine, guanine, uracil, cytosine, or thymine. In some embodiments, the modified nucleobase mimics the spatial arrangement, electronic properties, or some other physicochemical properties of the nucleobase and possesses the property of hydrogen bonding that binds one nucleic acid strand to another in a sequence-specific manner. In some embodiments, the modified nucleobase may pair with all five naturally occurring bases (uracil, thymine, adenine, cytosine, or guanine) without substantially affecting melting behavior, recognition by intracellular enzymes, or the activity of the oligonucleotide duplex. As used herein, the term “nucleobase” also includes structural analogs used in place of natural or naturally occurring nucleotides, such as the modified nucleobase and nucleobase analogs.In some embodiments, the nucleobase is an optionally substituted A, T, C, G, or U, or an optionally substituted tautomer of A, T, C, G, or U. In some embodiments, "nucleobase" refers to a nucleobase unit in an oligonucleotide or nucleic acid (e.g., A, T, C, G, or U as in an oligonucleotide or nucleic acid).

[0112] Nucleoside: The term "nucleoside" refers to a moiety in which a nucleobase or a modified nucleobase is covalently bonded to a sugar or a modified sugar. In some embodiments, the nucleoside is a natural nucleoside, e.g., adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, or deoxycytidine. In some embodiments, the nucleoside is a modified nucleoside, e.g., a substituted natural nucleoside (selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine). In some embodiments, the nucleoside is a substituted tautomer of a natural nucleoside selected from adenosine, deoxyadenosine, guanosine, deoxyguanosine, thymidine, uridine, cytidine, and deoxycytidine. In some embodiments, "nucleoside" refers to a nucleoside unit in an oligonucleotide or nucleic acid.

[0113] Nucleotide: As used herein, the term "nucleotide" refers to a monomeric unit of a polynucleotide composed of a nucleobase, a sugar, and one or more internucleotide links (e.g., phosphate links in natural DNA and RNA). It should be understood that naturally occurring bases [guanine (G), adenine (A), cytosine (C), thymine (T), and uracil (U)] are derivatives of purines or pyrimidines, but include naturally occurring or non-naturally occurring base analogs. It should be understood that naturally occurring sugars are pentose (pentose sugars) deoxyribose (which forms DNA) or ribose (which forms RNA), but include naturally occurring or non-naturally occurring sugar analogs. Nucleotides are linked through internucleotide links to form nucleic acids or polynucleotides. Many internucleotide links (such as, but not limited to, phosphates, phosphorothioates, boranophosphates, etc.) are known in the art. Artificial nucleic acids include PNA (peptide nucleic acid), phosphotriesters, phosphorothionates, H-phosphonates, phosphoamidates, boranophosphates, methylphosphonates, phosphonoacetates, thiophosphonoacetates, and other variants of the phosphate backbone of natural nucleic acids, such as those described herein. In some embodiments, natural nucleotides comprise naturally occurring bases, sugars, and linkages between nucleotides. As used herein, the term “nucleotide” also includes structural analogs used in place of natural or naturally occurring nucleotides, such as modified nucleotides and nucleotide analogs. In some embodiments, “nucleotide” refers to a nucleotide unit in an oligonucleotide or nucleic acid.

[0114] Oligonucleotide: The term "oligonucleotide" refers to a polymer or oligomer of nucleotides and may include any combination of natural and non-natural nucleobases, sugars, and linkages between nucleotides.

[0115] Oligonucleotides may be single-stranded or double-stranded. Single-stranded oligonucleotides may have a double-stranded region (formed by two parts of the single-stranded oligonucleotide), and double-stranded oligonucleotides comprising two oligonucleotide chains may have a single-stranded region, for example, in a region where the two oligonucleotide chains are not complementary to each other. Exemplary oligonucleotides include, but are not limited to, structural genes, genes containing control and termination regions, self-replicating systems such as viral or plasmid DNA, single-stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNA, microRNA mimics, supermir, aptamer, antimir, antagomir, U1 adapter, triplex-forming oligonucleotides, G-quadruplex oligonucleotides, RNA activators, immunostimulatory oligonucleotides, and decoy oligonucleotides.

[0116] The oligonucleotides of the present disclosure may have various lengths. In certain embodiments, the oligonucleotides may be in the range of about 2 to about 200 nucleosides in length. In various related embodiments, single-stranded, double-stranded, or triple-stranded oligonucleotides may be in the range of about 4 to about 10 nucleosides, about 10 to about 50 nucleosides, about 20 to about 50 nucleosides, about 15 to about 30 nucleosides, or about 20 to about 30 nucleosides in length. In some embodiments, the oligonucleotides are about 9 to about 39 nucleosides in length. In some embodiments, the oligonucleotides are about 25 to about 70 nucleosides in length. In some embodiments, the oligonucleotides are about 26 to about 70 nucleosides in length. In some embodiments, the oligonucleotides are about 27 to about 70 nucleosides in length. In some embodiments, the oligonucleotide is about 28 to about 70 nucleosides long. In some embodiments, the oligonucleotide is about 29 to about 70 nucleosides long. In some embodiments, the oligonucleotide is about 30 to about 70 nucleosides long. In some embodiments, the oligonucleotide is about 31 to about 70 nucleosides long. In some embodiments, the oligonucleotide is about 32 to about 70 nucleosides long. In some embodiments, the oligonucleotide is about 25 to about 60 nucleosides long. In some embodiments, the oligonucleotide is about 25 to about 50 nucleosides long. In some embodiments, the oligonucleotide is about 25 to about 40 nucleosides long. In some embodiments, the oligonucleotide is about 30 to about 40 nucleosides long.In some embodiments, the oligonucleotide is at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleosides long. In some embodiments, the oligonucleotide is at least 4 nucleosides long. In some embodiments, the oligonucleotide is at least 5 nucleosides long. In some embodiments, the oligonucleotide is at least 6 nucleosides long. In some embodiments, the oligonucleotide is at least 7 nucleosides long. In some embodiments, the oligonucleotide is at least 8 nucleosides long. In some embodiments, the oligonucleotide is at least 9 nucleosides long. In some embodiments, the oligonucleotide is at least 10 nucleosides long. In some embodiments, the oligonucleotide is at least 11 nucleosides long. In some embodiments, the oligonucleotide is at least 12 nucleosides long. In some embodiments, the oligonucleotide is at least 15 nucleosides long. In some embodiments, the oligonucleotide is at least 15 nucleosides long. In some embodiments, the oligonucleotide is at least 16 nucleosides long. In some embodiments, the oligonucleotide is at least 17 nucleosides long. In some embodiments, the oligonucleotide is at least 18 nucleosides long. In some embodiments, the oligonucleotide is at least 19 nucleosides long. In some embodiments, the oligonucleotide is at least 20 nucleosides long. In some embodiments, the oligonucleotide is at least 25 nucleosides long. In some embodiments, the oligonucleotide is at least 26 nucleosides long. In some embodiments, the oligonucleotide is at least 27 nucleosides long. In some embodiments, the oligonucleotide is at least 28 nucleosides long. In some embodiments, the oligonucleotide is at least 29 nucleosides long.In some embodiments, the oligonucleotide is at least 30 nucleosides long. In some embodiments, the oligonucleotide is at least 31 nucleosides long. In some embodiments, the oligonucleotide is at least 32 nucleosides long. In some embodiments, the oligonucleotide is at least 33 nucleosides long. In some embodiments, the oligonucleotide is at least 34 nucleosides long. In some embodiments, the oligonucleotide is at least 35 nucleosides long. In some embodiments, the oligonucleotide is at least 36 nucleosides long. In some embodiments, the oligonucleotide is at least 37 nucleosides long. In some embodiments, the oligonucleotide is at least 38 nucleosides long. In some embodiments, the oligonucleotide is at least 39 nucleosides long. In some embodiments, the oligonucleotide is at least 40 nucleosides long. In some embodiments, the oligonucleotide is 25 nucleosides long. In some embodiments, the oligonucleotide is 26 nucleosides long. In some embodiments, the oligonucleotide is 27 nucleosides long. In some embodiments, the oligonucleotide is 28 nucleosides long. In some embodiments, the oligonucleotide is 29 nucleosides long. In some embodiments, the oligonucleotide is 30 nucleosides long. In some embodiments, the oligonucleotide is 31 nucleosides long. In some embodiments, the oligonucleotide is 32 nucleosides long. In some embodiments, the oligonucleotide is 33 nucleosides long. In some embodiments, the oligonucleotide is 34 nucleosides long. In some embodiments, the oligonucleotide is 35 nucleosides long. In some embodiments, the oligonucleotide is 36 nucleosides long. In some embodiments, the oligonucleotide is 37 nucleosides long. In some embodiments, the oligonucleotide is 38 nucleosides long.In some embodiments, the oligonucleotide is 39 nucleosides long. In some embodiments, the oligonucleotide is 40 nucleosides long. In some embodiments, each nucleoside counted in the oligonucleotide length independently comprises a nucleobase comprising a ring having at least one nitrogen ring atom. In some embodiments, each nucleoside counted in the oligonucleotide length independently comprises A, T, C, G, or U, or optionally substituted A, T, C, G, or U, or optionally substituted tautomers of A, T, C, G, or U.

[0117] Optionally Substituted: As described herein, compounds of the present disclosure may comprise optionally substituted moiety, substituted moiety and / or unsubstituted moiety. Generally, the term “substituted” means that one or more hydrogens of a designated moiety are independently replaced by substituents. Unless otherwise specified, the “optionally substituted” group may independently have substituents at each substitutable position of the group, and if two or more positions within any given structure may be substituted by two or more substituents, the substituents may be the same or different at all positions. In some embodiments, the optionally substituted group is not substituted. In some embodiments, the optionally substituted group is substituted. Various substituents are listed below.

[0118] Monovalent substituents are independently halogens; -(CH2) 0-4 R°; -(CH2) 0-4 OR°; -O(CH2) 0-4 R°, -O-(CH2) 0-4 C(O)OR°; -(CH2) 0-4 CH(OR°)2; -(CH2) 0-4 Ph(can be substituted with R°); -(CH2) 0-4 O(CH2) 0-1Ph(can be replaced with R°); -CH=CHPh(can be replaced with R°); -(CH2) 0-4 O(CH2) 0-1 -pyridyl(can be substituted with R°); -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R°)2; -(CH2) 0-4 N(R°)C(O)R°; -N(R°)C(S)R°; -(CH2) 0-4 N(R°)C(O)N(R°)2; -N(R°)C(S)N(R°)2; -(CH2) 0-4 N(R°)C(O)OR°; -N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)N(R°)2; -N(R°)N(R°)C(O)OR°; -(CH2) 0-4 C(O)R°; -C(S)R°; -(CH2) 0-4 C(O)OR°; -(CH2) 0-4 C(O)SR°; -(CH2) 0-4 C(O)OSi(R°)3; -(CH2) 0-4 OC(O)R°; -OC(O)(CH2) 0-4 SR°, -SC(S)SR°; -(CH2) 0-4 SC(O)R°; -(CH2) 0-4 C(O)N(R°)2; -C(S)N(R°)2; -C(S)SR°; -SC(S)SR°, -(CH2) 0-4 OC(O)N(R°)2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O)R°; -C(NOR°)R°; -(CH2) 0-4 SSR°; -(CH2) 0-4 S(O)2R°; -(CH2) 0-4 S(O)2OR°; -(CH2) 0-4 OS(O)2R°; -S(O)2N(R°)2; -(CH2) 0-4S(O)R°; -N(R°)S(O)2N(R°)2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NH)N(R°)2; -Si(R°)3; -OSi(R°)3; -P(R°)2; -P(OR°)2; -OP(R°)2; -OP(OR°)2; -N(R°)P(R°)2; -B(R°)2; -OB(R°)2; -P(O)(R°)2; -OP(O)(R°)2; -N(R°)P(O)(R°)2; -(C 1-4 Straight-chain or branched alkylene)ON(R°)2; or -(C 1-4 (Straight-chain or branched alkylene)C(O)ON(R°)2; where each R° can be independently substituted as defined below and independently hydrogen, C 1-10 (e.g., C 1-6 , C 1-4 C having 1 to 5 heteroatoms independently selected from aliphatic, nitrogen, oxygen, sulfur, silicon, and phosphorus. 1-10 (e.g., C 1-6 , C 1-4 et al.) Heterolipids, C 6-10 (e.g., C6, C 10 5-10 (e.g., 5-9, 5-6, 5, 6, 9, 10, etc.) aryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, -CH2-(C 6-10 (e.g., C6, C 10 etc.) Aryl), -O(CH2) 0-1 (C 6-10 (e.g., C6, C 10 -CH2- (5-10 (e.g., 5-9, 5-6, 5, 6, 9, 10, etc.) heteroaryl having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur), -O(CH2) having 1-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur 0-1(a 5–10 (e.g., 5–9, 5–6, 5, 6, 9, 10, etc.) cyclic heteroaryl, a 3–10 (e.g., 3–6, 5–6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) cyclic, monocyclic, dicyclic, or polycyclic, saturated or partially unsaturated ring having 0–5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, or, notwithstanding the definition above, taking together two independent entities of R° and the atom(s) interposed between them, a 3–10 (e.g., 3–6, 5–6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) cyclic monocyclic ring having 0–5 heteroatoms independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus in addition to the interposed atom(s), It forms a dicyclic or polycyclic, saturated, partially unsaturated, or aromatic ring (in the case of an aromatic ring, 5 to 10 (e.g., 5 to 9, 5 to 6, 5, 6, 9, 10, etc.) ring), which can be substituted as defined below.

[0119] The monovalent substituent of R° (or a ring formed by taking two independent instances of R° and the atom interposed between them) is independently a halogen, -(CH2) 0-2 R ● , -(HalloR ● ), -(CH2) 0-2 OH, -(CH2) 0-2 OR ● , -(CH2) 0-2 CH(OR ● )2; -O(HalloR ● ), -CN, -N3, -(CH2) 0-2 C(O)R ● , -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● , -(CH2) 0-2 SR ● , -(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR ● , -(CH2)0-2 NR ● 2, -NO2, -SiR ● 3, -OSiR ● 3, -C(O)SR ● , -(C 1-4 Straight-chain or branched alkylene)C(O)OR ● , or -SSR ● and here, each R ● is unsubstituted or, when preceded by "halo", is substituted with only one or more halogens, and independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 pH is selected from a 3- to 6 (e.g., 3-5, 5-6, etc.) saturated, partially unsaturated, or aromatic ring (in the case of an aromatic ring, 5 or 6 members) having 0- to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. The divalent substituent on the saturated carbon atom of R° is independently =O or =S.

[0120] The divalent substituents are independently as follows: =O, =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2-3 O- or -S(C(R * 2)) 2-3 S-, here R * Each independent entity of can be substituted with hydrogen, C as defined below. 1-6 It is selected from aliphatic, or unsubstituted 3–6 (e.g., 3–5, 5–6, etc.) saturated, partially unsaturated, or aromatic rings (in the case of aromatic rings, 5 or 6 members) having 0–4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. The divalent substituent bonded to the vinyl-substitutable carbon of the "optionally substituted" group is independently -O(CR *2) 2-3 O- and, where R * Each independent entity of can be substituted with hydrogen, C as defined below. 1-6 It is selected from aliphatic, or unsubstituted 3 to 6 (e.g., 3 to 5, 5 to 6, etc.) saturated, partially unsaturated, or aromatic rings (in the case of aromatic rings, 5 or 6) having 0 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0121] R * The substituents on the aliphatic phase of are independently halogens, -R ● , -(HalloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2 or -NO2, where each R ● It is unsubstituted, or if preceded by "halo," it is substituted with only one or more halogens, and independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 It is a 3- to 6 (e.g., 3-5, 5-6, etc.) saturated, partially unsaturated, or aromatic ring (in the case of an aromatic ring, a 5- or 6-membered ring) having 0- to 4 heteroatoms independently selected from pH, or nitrogen, oxygen, and sulfur.

[0122] Substitutable substituents on the nitrogen phase are independently -R † , -NR † 2, -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH2C(O)R † , -S(O)2R † , -S(O)2NR † 2, -C(S)NR † 2, -C(NH)NR † 2 or -N(R † )S(O)2R† and; here, each R † C, which can be independently substituted for hydrogen as defined below. 1-6 Aliphatic, unsubstituted -OPh, or unsubstituted 3 to 6 (e.g., 3 to 5, 5 to 6, etc.) saturated, partially unsaturated, or aromatic rings (in the case of aromatic rings, 5 or 6), having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or notwithstanding the definition above, two independent R † The presence of [the element], along with the interposed atoms, forms an unsubstituted 3 to 12 (e.g., 3 to 10, 3 to 6, 5 to 10, 5 to 6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) saturated, partially unsaturated, or aryl monocyclic or dicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0123] R † The substituents on the aliphatic phase of are independently halogens, -R ● , -(HalloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2 or -NO2, where each R ● It is unsubstituted, or if preceded by "halo," it is substituted with only one or more halogens, and independently C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 It is a 3- to 6 (e.g., 3-5, 5-6, etc.) saturated, partially unsaturated, or aromatic ring (in the case of an aromatic ring, a 5- or 6-membered ring) having 0- to 4 heteroatoms independently selected from pH, or nitrogen, oxygen, and sulfur.

[0124] Partially unsaturated: As used herein, the term “partially unsaturated” refers to a ring moiety comprising at least one double or triple bond. The term “partially unsaturated” is intended to include a ring having multiple unsaturated sites, but is not intended to include an aryl or heteroaryl moiety as defined herein.

[0125] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in a unit dose amount appropriate for administration in a therapeutic regimen that represents a statistically significant probability of achieving a desired therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated to be administered in solid or liquid form, including those suitable for: oral administration, e.g., drench (aqueous or non-aqueous solution or suspension), tablets (e.g., buccal, sublingual, and intended for systemic absorption), bolus, powder, granule, paste for application to the tongue; parenteral administration, e.g., sterile solution or suspension, or sustained-release formulation, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection; topical application, e.g., cream, ointment, or controlled-release patch or spray applied to the skin, lungs, or oral cavity; vaginal or rectal administration, e.g., pessary, cream, or foam; sublingual; Eyes; skin; or nasal cavity, lungs, and other mucosal surfaces.

[0126] Pharmaceutically acceptable: As used herein, the phrase “pharmaceutically acceptable” refers to the applicable compounds, substances, compositions, and / or dosage forms suitable for use in contact with human and animal tissues corresponding to a reasonable benefit / risk ratio, without excessive toxicity, irritation, allergic reaction, or other problems or complications, within the scope of sound medical judgment.

[0127] Pharmaceutically Acceptable Carriers: As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable substance, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, that is involved in carrying or transporting a target compound from one organ or part of the body to another organ or part of the body. Each carrier must be “acceptable” in terms of compatibility with other components of the formulation and not harmful to the patient. Some examples of substances that may be used as pharmaceutically acceptable carriers include: sugars, e.g., lactose, glucose, sucrose; starches, e.g., corn starch and potato starch; cellulose, and derivatives thereof, e.g., sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanths; malt; gelatin; talc; excipients, e.g., cocoa butter and suppository wax; oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; Glycols, e.g., propylene glycol; polyols, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol; esters, e.g., ethyl oleate and ethyl laurate; agar; buffers, e.g., magnesium hydroxide and aluminum hydroxide; alginic acid; water free of pyrogens; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solution; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic compatible materials used in pharmaceutical formulations.

[0128] Pharmaceutically Acceptable Salt: As used herein, the term “pharmaceutically acceptable salt” refers to a salt of such compound suitable for use in a pharmaceutical context, that is, a salt suitable for use in contact with human and subanimal tissues corresponding to a reasonable benefit / risk ratio, within the scope of sound medical judgment, without excessive toxicity, irritation, allergic reaction, etc. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts of amino groups formed by inorganic acids, e.g., hydrochloric acid, hydrobromide, phosphoric acid, sulfuric acid, and perchloric acid, or by organic acids, e.g., acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, e.g., ion exchange.In some embodiments, pharmaceutically acceptable salts are adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, maleate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, palmoate, Pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. are included, but not limited thereto. In some embodiments, the provided compound comprises one or more acidic groups, e.g., oligonucleotides, and the pharmaceutically acceptable salt is an alkali, alkaline earth metal, or ammonium salt (e.g., an ammonium salt of N(R)3, where each R is independently defined and described in this disclosure). Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. In some embodiments, the pharmaceutically acceptable salt is a sodium salt. In some embodiments, the pharmaceutically acceptable salt is a potassium salt. In some embodiments, the pharmaceutically acceptable salt is a calcium salt. In some embodiments, the pharmaceutically acceptable salt comprises, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and aryl sulfonates.In some embodiments, the provided compound comprises one or more acid groups, and, for example, the oligonucleotide may comprise two or more acid groups (e.g., in natural phosphate linkages and / or modified internucleotide linkages). In some embodiments, the pharmaceutically acceptable salt of such compound, or generally the salt, comprises two or more cations, which may be the same or different. In some embodiments, in the pharmaceutically acceptable salt (or generally the salt), all ionizable hydrogens (e.g., in an aqueous solution where the pKa is about 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2 or less; in some embodiments, about 7 or less; in some embodiments, about 6 or less; in some embodiments, about 5 or less; in some embodiments, about 4 or less; in some embodiments, about 3 or less) are replaced by cations in the acid groups. In some embodiments, each phosphorothioate and phosphate group exists independently in its salt form (e.g., -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively, in the case of a sodium salt). In some embodiments, the link between each phosphorothioate and phosphate nucleotide exists independently in its salt form (e.g., -OP(O)(SNa)-O- and -OP(O)(ONa)-O-, respectively, in the case of a sodium salt). In some embodiments, the pharmaceutically acceptable salt is the sodium salt of the oligonucleotide. In some embodiments, the pharmaceutically acceptable salt is the sodium salt of the oligonucleotide, wherein each acidic phosphate and modified phosphate group (e.g., phosphorothioate, phosphate, etc.) exists in salt form where present (all sodium salts).

[0129] Determined: Determined (or pre-determined) means intentionally selected, not random, or controlled, as opposed to, for example, occurring randomly, random, or achieved without control. Those skilled in the art will understand from reading this specification that the present disclosure provides a technique that enables the selection of specific chemical and / or stereochemical features to be incorporated into an oligonucleotide composition, and further enables the controlled preparation of an oligonucleotide composition having such chemical and / or stereochemical features. Such provided compositions are "determined" as described herein. A composition that may contain a specific oligonucleotide is not a "determined" composition because it was created by chance through an uncontrolled process to intentionally generate a specific chemical and / or stereochemical feature. In some embodiments, the predetermined composition is a composition that can be intentionally reproduced (e.g., through the repetition of a controlled process). In some embodiments, a predetermined level of a plurality of oligonucleotides in the composition means that the absolute amount and / or relative amount (ratio, percentage, etc.) of the plurality of oligonucleotides in the composition is controlled. In some embodiments, a predetermined level of a plurality of oligonucleotides in the composition is achieved through chiral-controlled oligonucleotide preparation.

[0130] Protecting Groups: As used herein, the term “protecting group” includes those well known in the art and described in detail in *Protecting Groups in Organic Synthesis*, TW Greene and PGM Wuts, 3rd edition, John Wiley & Sons, 1999, the whole of which is incorporated herein by reference. It also includes protecting groups specifically applied to nucleoside and nucleotide chemistry as described in *Current Protocols in Nucleic Acid Chemistry*, edited by Serge L. Beaucage et al., June 2012, the whole of Chapter 2 of which is incorporated herein by reference. Suitable amino-protecting groups include methyl carbamate, ethyl carbamante, 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), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), and 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-DicyclohexylCarboxamido)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 dithiocarbamate, Benzyl carbamate (Cbz), p-Methoxybenzyl carbamate (Moz), p-Nitobenzyl 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, [2-(1,3-Dithianyl)]methyl carbamate (Dmoc), 4-Methylthiophenyl carbamate (Mtpc), 2,4-Dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl derivative, N'-p-toluenesulfonylaminocarbonyl derivative, N'-phenylaminothiocarbonyl derivative, t-amyl Carbamate, S-Benzyl Thiocarbamate, p-Cyanobenzyl Carbamate, Cyclobutyl Carbamate, Cyclohexyl Carbamate, Cyclopentyl Carbamate, Cyclopropylmethyl Carbamate, p-Decyloxybenzyl Carbamate, 2,2-Dimethoxycarbonylvinyl Carbamate, o-(N,N-DimethylCarboxamido)Benzyl Carbamate, 1,1-Dimethyl-3-(N,N-DimethylCarboxamido)propyl Carbamate, 1,1-Dimethylpropynyl Carbamate, Di(2-Pyridyl)methyl Carbamate, 2-Furanylmethyl Carbamate, 2-Iodoethyl Carbamate, Isovorinyl Carbamate, Isobutyl Carbamate, Isonicotinyl Carbamate,p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-Pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrosinnaamide, N-acetylmethionine derivative, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisiliazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazcyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazcyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrroline-3-yl)amine, quaternary ammonium salt, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine,N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, Np-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)methyl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivative, N-diphenylboric acid derivative, N-[phenyl(pentacarbonylchrome- or tungsten)carbonyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidate, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys), p-Toluenesulfonamide (Ts), Benzenesulfonamide, 2,3,6-Trimethyl-4-Methoxybenzenesulfonamide (Mtr), 2,4,6-Trimethoxybenzenesulfonamide (Mtb), 2,6-Dimethyl-4-Methoxybenzenesulfonamide (Pme), 2,3,5,6-Tetramethyl-4-Methoxybenzenesulfonamide (Mte), 4-Methoxybenzenesulfonamide (Mbs), 2,4,6-Trimethylbenzenesulfonamide (Mts), 2,6-Dimethoxy-4-Methylbenzenesulfonamide (iMds), 2,2,5,7,8-Pentamethylchroman-6-Sulfamide (Pmc), Methanesulfonamide (Ms), β-Trimethylsilylethanesulfonamide (SES), 9-Anthracenesulfonamide, 4-(4',Includes 8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0131] Suitablely reported carboxylic acids further include, but are not limited to, silyl-, alkyl-, alkenyl-, aryl-, and arylalkyl-protected carboxylic acids. Examples of suitable silyl groups include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, etc. Examples of suitable alkyl groups include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, tetrahydropyran-2-yl. Examples of suitable alkenyl groups include allyl. Examples of suitable aryl groups include optionally substituted phenyl, biphenyl, or naphthyl. Examples of suitable arylalkyl groups include optionally substituted benzyls (e.g., p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl), and 2- and 4-picolyls.

[0132] Suitable hydroxyl 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, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranil (THP), 3-bromotetrahydropyranil, tetrahydrothiopyranil, 1-methoxycyclohexyl, 4-methoxytetrahydropyranil (MTHP), 4-Methoxytetrahydrothiopyranil, 4-Methoxytetrahydrothiopyranil S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxane-2-yl, tetrahydrofuranil, tetrahydrothiopyranil, 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, 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-oxydo, diphenylmethyl, p,p'-dinitrobenzylhydryl, 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromopenacyloxyphenyl)diphenylmethyl, 4,4',4''-Tris(4,5-Dichlorophthalimidophenyl)methyl, 4,4',4''-Tris(levulinoyloxyphenyl)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)xantenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxid, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethyltexylsilyl, 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-Chlorophenoacetate, 3-Phenylpropionate, 4-Oxopentanoate (Rebulinate), 4,4-(Ethylenedithio)pentanoate (Rebulinoyldithioacetal), Pivaloate, Adamantoate, Crotonate, 4-Methoxycrotonate, Benzoate, p-Phenylbenzoate, 2,4,6-Trimethylbenzoate (Methitoate), Alkyl methyl carbonate, 9-Fluorenylmethyl carbonate (Fmoc), Alkyl ethyl carbonate, Alkyl 2,2,2-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 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-(methoxycarbonyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, It includes dimethylphosphinothioryl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). To protect 1,2- or 1,3-diols, the protecting group is methylene acetal, ethylidene acetal, 1-t-butylethylidene ketal, 1-phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene ortho ester, 1-methoxyethylidene ortho ester, 1-ethoxyethylidene ortho ester, Includes 1,2-dimethoxyethylidene ortho esters, α-methoxybenzylidene ortho esters, 1-(N,N-dimethylamino)ethylidene derivatives, α-(N,N'-dimethylamino)benzylidene derivatives, 2-oxacyclopentylidene ortho esters, di-t-butylsilylene groups (DTBS), 1,3-(1,1,3,3-tetraisopropyldisilosanilidene) derivatives (TIPDS), tetra-t-butoxydisiloxane-1,3-diylidene derivatives (TBDS), cyclic carbonates, cyclic boronates, ethyl boronates, and phenyl boronates.

[0133] In some embodiments, the hydroxyl protecting group is acetyl, t-butyl, t-butoxymethyl, methoxymethyl, tetrahydropyranyl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 2-trimethylsilylethyl, p-chlorophenyl, 2,4-dinitrophenyl, benzyl, benzoyl, p-phenylbenzoyl, 2,6-dichlorobenzyl, diphenylmethyl, p-nitrobenzyl, triphenylmethyl(trityl), 4,4'-dimethoxytrityl, trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triphenylsilyl, triisopropylsilyl, benzoyl formate, chloroacetyl, trichloroacetyl, trifluoroacetyl, pivaloyl, 9-fluorenylmethyl carbonate, mesylate, tosylate, tritriplate, trityl, Monomethoxytrityl (MMTr), 4,4'-Dimethoxytrityl (DMTr) and 4,4',4''-Trimethoxytrityl (TMTr), 2-Cyanoethyl (CE or Cne), 2-(Trimethylsilyl)ethyl (TSE), 2-(2-Nitrophenyl)ethyl, 2-(4-Cyanophenyl)ethyl, 2-(4-Nitrophenyl)ethyl (NPE), 2-(4-Nitrophenylsulfonyl)ethyl, 3,5-Dichlorophenyl, 2,4-Dimethylphenyl, 2-Nitrophenyl, 4-Nitrophenyl, 2,4,6-Trimethylphenyl, 2-(2-Nitrophenyl)ethyl, Butylthiocarbonyl, 4,4',4''-Tris(Benzoyloxy)trityl, Diphenylcarbamoyl, Levulinyl, 2-(Dibromomethyl)benzoyl (Dbmb), It is 2-(isopropylthiomethoxymethyl)benzoyl (Ptmt), 9-phenylxanthen-9-yl (Pixyl), or 9-(p-methoxyphenyl)xanthen-9-yl (MOX). In some embodiments, each hydroxyl protecting group is independently selected from acetyl, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, and 4,4'-dimethoxycitriteyl. In some embodiments, the hydroxyl protecting group is selected from the group consisting of trityl, monomethoxycitriteyl, and 4,4'-dimethoxycitriteyl groups. In some embodiments, the phosphate linkage protecting group is a group attached to the phosphate linkage (e.g., internucleotide linkage) throughout the oligonucleotide synthesis. In some embodiments, the protecting group is attached to the sulfur atom of the phosphorothioate group.In some embodiments, the protecting group is attached to the oxygen atom of the phosphorothioate link between nucleotides. In some embodiments, the protecting group is attached to the oxygen atom of the phosphate link between nucleotides. In some embodiments, the protecting group is 2-cyanoethyl (CE or Cne), 2-trimethylsilylethyl, 2-nitroethyl, 2-sulfonylethyl, methyl, benzyl, o-nitrobenzyl, 2-(p-nitrophenyl)ethyl (NPE or Npe), 2-phenylethyl, 3-(N-tert-butylcarboxamido)-1-propyl, 4-oxopentyl, 4-methylthio-1-butyl, 2-cyano-1,1-dimethylethyl, 4-N-methylaminobutyl, 3-(2-pyridyl)-1-propyl, 2-[N-methyl-N-(2-pyridyl)]aminoethyl, 2-(N-formyl,N-methyl)aminoethyl, or 4-[N-methyl-N-(2,2,2-trifluoroacetyl)amino]butyl.

[0134] Subject: As used herein, the term “subject” or “test subject” refers to any organism to which a compound (e.g., oligonucleotide) or composition is administered in accordance with this disclosure, for example, for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals, e.g., mice, rats, rabbits, non-human primates, and humans; insects; worms; etc.) and plants. In some embodiments, the subject is a human. In some embodiments, the subject may have or be susceptible to a disease, disorder, and / or pathological condition.

[0135] Substantially: As used herein, the term “substantially” refers to a qualitative condition indicating the full or nearly full range or degree of a characteristic or property of interest. A base sequence that is substantially identical or complementary to a second sequence is not completely identical or complementary to the second sequence, but is mostly identical or nearly identical or complementary to the second sequence. In some embodiments, an oligonucleotide having a sequence substantially complementary to another oligonucleotide or nucleic acid forms a duplex with the oligonucleotide or nucleic acid in a manner similar to that of an oligonucleotide having a completely complementary sequence. Furthermore, a person skilled in the art of biology and / or chemistry will understand that biological and chemical phenomena rarely complete or / or never proceed toward completeness, or achieve or avoid absolute results. Accordingly, the term “substantially” is used herein to capture the potential lack of completeness inherent in many biological and / or chemical phenomena.

[0136] Sugar: The term “sugar” refers to a monosaccharide or polysaccharide in a closed and / or open form. In some embodiments, the sugar is a monosaccharide. In some embodiments, the sugar is a polysaccharide. The sugar includes, but is not limited to, ribose, deoxyribose, pentopuranose, pentopyranoose, and hexopyranoose moiety. As used herein, the term “sugar” also includes structural analogs used in place of conventional sugar molecules, such as glycols (polymers thereof form the backbone of nucleic acid analogs), glycol nucleic acids (“GNA”), etc. As used herein, the term “sugar” also includes structural analogs used in place of natural or naturally occurring nucleotides, such as modified sugars and nucleotide sugars. In some embodiments, the sugar is an RNA or DNA sugar (ribose or deoxyribose). In some embodiments, the sugar is a modified ribose or deoxyribose sugar, such as a 2’-modified, 5’-modified, etc. As described herein, in some embodiments, the modified sugar, when used in an oligonucleotide and / or nucleic acid, may provide one or more desired properties, activities, etc. In some embodiments, the sugar is optionally substituted ribose or deoxyribose. In some embodiments, "sugar" refers to a sugar unit in an oligonucleotide or nucleic acid.

[0137] Predisposed: An individual "predisposed" to a disease, disorder, and / or condition is an individual whose risk of developing that disease, disorder, and / or condition is higher than that of members of the general public. In some embodiments, an individual predisposed to a disease, disorder, and / or condition has a predisposition to have that disease, disorder, and / or condition. In some embodiments, an individual predisposed to a disease, disorder, and / or condition may not have ever been diagnosed with that disease, disorder, and / or condition. In some embodiments, an individual predisposed to a disease, disorder, and / or condition may exhibit symptoms of that disease, disorder, and / or condition. In some embodiments, an individual predisposed to a disease, disorder, and / or condition may not exhibit symptoms of that disease, disorder, and / or condition. In some embodiments, an individual predisposed to a disease, disorder, and / or condition will develop the disease, disorder, and / or condition. In some embodiments, an object susceptible to disease, disability, and / or pathological condition will not develop disease, disability, and / or pathological condition.

[0138] Therapeutic: As used herein, the term “therapeutic” generally refers to any preparation that, when administered to a subject, produces a desired effect (e.g., a desired biological, clinical, or pharmacological effect). In some embodiments, a preparation is considered a therapeutic if it produces a statistically significant effect over an appropriate population. In some embodiments, the appropriate population is a group of subjects who have or are susceptible to a disease, disorder, or pathology. In some embodiments, the appropriate population is a group of model organisms. In some embodiments, the appropriate population may be defined by one or more criteria such as age group, sex, genetic background, pre-existing clinical condition, or prior exposure to a therapy. In some embodiments, the therapeutic is a substance that, when administered to a subject in an effective amount, alleviates, improves, mitigates, suppresses, prevents, delays, reduces, or decreases the severity of one or more symptoms or features of a disease, disorder, and / or pathology in the subject. In some embodiments, the “therapeutic agent” is a drug that has been approved by or is required to be approved by a government agency before it can be sold for human administration. In some embodiments, the “therapeutic agent” is a drug that requires a medical prescription for administration to humans. In some embodiments, the therapeutic agent is a provided compound, e.g., a provided oligonucleotide.

[0139] Therapeutic Effective Dose: As used herein, the term “therapeutic effective dose” means an amount of a substance (e.g., therapeutic agent, composition, and / or formulation) that, when administered as part of a therapeutic regimen, elicits a desired biological response. In some embodiments, the therapeutic effective dose of a substance is an amount sufficient to treat, diagnose, prevent, and / or delay the onset of a disease, disorder, and / or condition when administered to a subject suffering from or susceptible to such disease, disorder, and / or condition. As will be known to those skilled in the art, the effective dose of a substance may vary depending on factors such as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective dose of a compound in a formulation for treating a disease, disorder, and / or condition is an amount that alleviates, improves, reduces, inhibits, prevents, delays the onset, reduces severity, and / or reduces the incidence of one or more symptoms or features of the disease, disorder, and / or condition. In some embodiments, the therapeutic effective dose is administered as a single dose; in some embodiments, multiple unit doses are required to deliver the therapeutic effective dose.

[0140] Treatment: As used herein, the terms “treat,” “treatment,” or “treating” refer to any method used to partially or completely alleviate, improve, reduce, inhibit, prevent, delay onset, reduce severity, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to subjects who do not exhibit signs of the disease, disorder, and / or condition. In some embodiments, treatment may be administered to subjects who exhibit only early signs of the disease, disorder, and / or condition, for example, to reduce the risk of developing pathology associated with the disease, disorder, and / or condition.

[0141] Unsaturated: As used herein, the term "unsaturated" means that the moiety has one or more unsaturated units.

[0142] As will be known to those skilled in the art, the methods and compositions described herein with respect to the provided compounds (e.g., oligonucleotides) are generally also applicable to pharmaceutically acceptable salts of such compounds.

[0143] Description of a specific implementation example

[0144] Specific implementation examples are listed below.

[0145] Above all, the present disclosure includes the recognition that traditional techniques for incorporating -P(O)(OH)2 groups into compounds, such as the use of protecting groups like -P(O)(OMe)2, -P(O)(OEt)2, etc., often involve deprotection conditions that can reduce manufacturing efficiency and / or increase manufacturing costs. For example, when incorporating -P(O)(OH)2 into an oligonucleotide using -P(O)(OMe)2, -P(O)(OEt)2, etc., additional steps and / or harsher reaction conditions may be required for the removal of the methyl or ethyl group, which can result in reduced manufacturing efficiency, reduced oligonucleotide yield and / or purity and / or increased costs compared to the production of a reference oligonucleotide without -P(O)(OH)2 (e.g., the same oligonucleotide except for the absence of -P(O)(OH)2).

[0146] In some embodiments, the present disclosure provides a technique for incorporating -P(O)(OH)2 into a compound, e.g., an oligonucleotide, which may provide higher yield, higher purity, higher efficiency, lower operational complexity, milder reaction conditions, shorter manufacturing time and / or lower cost compared to using, e.g., -P(O)(OMe)2 or -P(O)(OEt)2. In some embodiments, the present disclosure provides a protected -P(O)(OH)2 group that can be deprotected under oligonucleotide synthesis conditions during cleavage and / or deprotection, which is utilized, e.g., for oligonucleotides lacking the -P(O)(OH)2 group. In some embodiments, the provided technique is useful for the preparation of stereorandom oligonucleotide compositions. In some embodiments, the provided technique is useful for stereoselective oligonucleotide synthesis. In some embodiments, stereoselective oligonucleotide synthesis produces each of a single stereochemical arrangement, which is one or more or the entire chiral linkage within the oligonucleotide, stereoselectively and independently of the others. In some embodiments, the provided technique is useful for producing chiral-controlled oligonucleotide compositions. In some embodiments, a chiral aid is utilized during stereoselective oligonucleotide synthesis. Specific useful stereoselective oligonucleotide synthesis techniques, including chiral aids, cycles, conditions, etc., are described in US 10167309, US 11643657, US 11718638, US 11608355, and US 20230089442, the full text of each of which is independently incorporated herein by reference.

[0147] For example, in some embodiments, the provided technology is used to prepare a compound containing an oligonucleotide as described herein -P(O)(OR PG Utilizes )2. In some implementations, -P(O)(OR PG)2 is converted to -P(O)(OH)2 during manufacturing or after administration to the subject.

[0148] In some embodiments, the present disclosure is -P(O)(OR PG It provides a technology including )2, wherein each R PG -P(O)(OH)2 is a group that is unstable under oligonucleotide synthesis conditions, for example, during cleavage, deprotection, and / or removal of chiral auxiliaries, which is utilized for reference oligonucleotides lacking -P(O)(OH)2. In some embodiments, each R PG is a group that can be removed under oligonucleotide synthesis conditions for a reference oligonucleotide that is free of the -P(O)(OH)2 group. In some embodiments, -P(O)(OR PG )2 is a protected -P(O)(OH)2 group that is converted to -P(O)(OH)2, which may exist in salt form under oligonucleotide synthesis conditions, for example. In some embodiments, as exemplified herein, the P atom is bonded to a carbon atom.

[0149] In some implementations, R PG is 1) an alpha carbon atom bonded to the oxygen to which it is attached (where the alpha carbon atom is bonded to hydrogen) and 2) a beta carbon atom (where the beta carbon atom is an electron-withdrawing group like R (e.g., compared to ethyl). PG R that can promote the removal of PG11 Includes (combined with). In some embodiments, R PG11 is an electronic absorber (e.g., described in US 20230089442). In some embodiments, R PG11 R as described herein s is. In some implementations, R PG11 is -CN. In some implementations, R PG11 -L s -R s11 and, here, each L s and R s11...is as independently described herein. In some embodiments, R PG is -CH2-R s and, where -CH2- is independently and optionally substituted, and R s is as described herein. In some embodiments, -CH2- is optionally a single substitution. In some embodiments, R PG R as described herein P1 is. In some implementations, R PG R as described herein P2 is. In some implementations, R PG is -O-CH2CH2CN.

[0150] In some implementations, R PG -CH2-O(CO)R PG12 and, here R PG12 is a trisubstituted methyl and -CH2- is optionally substituted. In some embodiments, R PG -CH2-O(CO)R PG12 and, here R PG12 is -C(R s11 )3 and here, each R s11 ...is as independently described herein. In some embodiments, R PG is R PG12 and, here R PG12 is a trisubstituted methyl. In some embodiments, R PG12 is -C(R s11 )3 and here, each R s11 ...is as independently described herein. In some embodiments, trisubstituted methyl or -C(R s11 The carbon of )3 is bonded to three carbon atoms. In some embodiments, R PG12 is an optionally substituted t-butyl. In some embodiments, R PG12 is t-butyl. In some embodiments, R PG R as described herein P1 or R P2 is. In some implementations, R PGis -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl. In some embodiments, R PG is -O-CH2-OC(O)-tBu.

[0151] In some implementations, -P(O)(OR PG )2 is -P(O)(R as described herein (e.g., in Formula I or Formula I'). P1 )(R P2 It has the structure of ), where R 1 and R P2 Each is as described independently in the present invention.

[0152] In some implementations, R PG is unstable under basic conditions. In some embodiments, R PG is unstable under acidic conditions. In some embodiments, -P(O)(OR PG )2 is converted to -P(O)(OH)2 under acidic conditions. In some embodiments, -P(O)(OR PG )2 is converted to -P(O)(OH)2 under basic conditions. In some embodiments, conditions are utilized in oligonucleotide synthesis for a reference oligonucleotide that does not have a -P(O)(OH)2 group but is otherwise identical. In some embodiments, conditions are cleavage and / or deprotection conditions in oligonucleotide synthesis. In some embodiments, conditions are chiral support removal, cleavage and / or deprotection conditions in oligonucleotide synthesis.

[0153] As recognized by those skilled in the art, acidic and / or basic groups, e.g., -P(O)(OH)2, may exist in various forms, including various salt forms. In some embodiments, acidic and / or basic groups, e.g., -P(O)(OH)2, exist in salt forms. In some embodiments, acidic and / or basic groups, e.g., -P(O)(OH)2, exist in pharmaceutically acceptable salt forms.

[0154] In some embodiments, as described below, the present disclosure provides, above all, techniques useful for synthesizing oligonucleotides, such as compounds (e.g., nucleosides, phosphoramidites, oligonucleotides, reaction reagents, etc.), methods, etc.

[0155] Phosphoramidite

[0156] Various phosphoramidites, such as those described in US 10167309, US 11643657, US 11718638, US 11608355, US 20230089442, etc., may be utilized for the production of oligonucleotides. In some embodiments, the present disclosure relates to -P(O)(OR) as described herein. PG The present invention provides a phosphoramidite comprising )2. In some embodiments, the phosphoramidite is -P(O)(OR )2 coupled to the sugar moiety of the nucleoside. PG Includes )2. In some embodiments, the phosphoramidite is attached to a moiety attached to a sugar moiety of the nucleoside, e.g., to an optionally substituted triazole ring -P(O)(OR PG Includes )2. In some embodiments, the sugar is a natural sugar. In some embodiments, the sugar is a modified sugar. In some embodiments, the nucleobase of the phosphoramidite is an optionally substituted natural nucleobase. In some embodiments, the nucleobase of the phosphoramidite is an optionally protected A, T, C, G, U, or 5mC (e.g., for oligonucleotide synthesis). In some embodiments, the phosphoramidite is baseless.

[0157] In some embodiments, the present disclosure is (R PG O)2P(O)-L 5E -SU(-BA)-OP(OR 1 )N(R 2 )(R 3 Provides a compound having the structure of ) or a salt thereof, wherein L 5Eis L as described herein, SU is a sugar as described herein, and each other variable is independently as described herein. In some embodiments, these compounds are utilized as phosphoramidites for oligonucleotide synthesis. In some embodiments, L 5E is an optionally substituted triazole ring.

[0158] In some embodiments, the provided technology provides a compound of formula P' or a salt thereof:

[0159] ,

[0160] P'

[0161] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized as phosphoramidites for oligonucleotide synthesis.

[0162] In some embodiments, the provided technology provides a compound of formula P or a salt thereof:

[0163] ,

[0164] P

[0165] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized as phosphoramidites for oligonucleotide synthesis.

[0166] In some embodiments, the provided technology provides a compound of formula I' or a salt thereof:

[0167] ,

[0168] I'

[0169] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized as phosphoramidites for oligonucleotide synthesis.

[0170] In some embodiments, the provided technology provides a compound of formula I or a salt thereof:

[0171] ,

[0172] I

[0173] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized as phosphoramidites for oligonucleotide synthesis.

[0174] Specific implementation examples of specific variables used in various chemical formulas are described below.

[0175] R P1

[0176] In some implementations, R P1 is -O-CH2CH2CN. In some implementations, R P1 -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl. In some embodiments, R P1 It is -O-CH2-OC(O)-tBu.

[0177] R P2

[0178] In some implementations, R P2 is -O-CH2CH2CN. In some implementations, R P2 -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl. In some embodiments, R P2 It is -O-CH2-OC(O)-tBu.

[0179] In some implementations, R P1 and R P2 is identical. In some implementations, R P1 and R P2 is different. In some implementations, R P1 and R P2 Each is -O-CH2CH2CN. In some implementations, R P1 and R P2Each independently -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl. In some embodiments, R P1 and R P2 Each is -O-CH2-OC(O)-tBu.

[0180] R P

[0181] In some implementations, R P is substituted tert-butyl. In some embodiments, R P is tert-butyl.

[0182] BA

[0183] In some embodiments, BA is hydrogen.

[0184] In some embodiments, BA is an optionally substituted 3 to 20 (e.g., 3 to 15, 3 to 10, 3 to 6, 5 to 10, 5 to 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, BA is an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) ring having 0 to 5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, BA is an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) ring having 0 to 5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from nitrogen and oxygen. In some embodiments, BA is an optionally substituted ring having at least one nitrogen atom. In some embodiments, BA is an optionally substituted 5 to 15 ring. In some embodiments, BA is an optionally substituted 5 to 10 ring. In some embodiments, BA is an optionally substituted 5- to 9-membered ring. In some embodiments, BA is an optionally substituted 9-membered ring. In some embodiments, BA is an optionally substituted 10-membered ring. In some embodiments, it is an optionally substituted 5-membered ring. In some embodiments, BA is an optionally substituted 6-membered ring.In some embodiments, BA is an optionally substituted monocyclic ring. In some embodiments, BA is an optionally substituted bicyclic ring. In some embodiments, BA is bonded to C1 of the sugar at the nitrogen atom.

[0185] In some embodiments, BA is an optionally substituted group, where the group is , , , , or It is either or its tautomer. In some embodiments, BA is an optionally protected nucleobase. In some embodiments, BA is a protected nucleobase. In some embodiments, BA is an optionally protected nucleobase selected from A, T, C, G, U, and 5mU. In some embodiments, the protection is suitable for oligonucleotide synthesis.

[0186] In some embodiments, BA is an optionally substituted or protected nucleobase. Various nucleobases are as described herein and may be used as BA.

[0187] SU

[0188] In some embodiments, SU is a sugar as described herein. In some embodiments, SU is And, where nucleobases, BA, etc. are bound to C1. In some embodiments, SU is And, where the nucleobase, BA, etc., is bound to C1 as described herein. In some embodiments, SU is optionally substituted And, here, nucleobases, BA, etc. are bound to C1 as described herein.

[0189] R 2s

[0190] In some implementations, R s2 is -H. In some implementations, R 2s is -F.

[0191] In some implementations, R 2sis -Cl. In some implementations, R 2s is -Br. In some implementations, R 2s is -I. In some implementations, R 2s is -CN. In some implementations, R 2s is -N3. In some implementations, R 2s is -NO. In some implementations, R 2s is -NO2.

[0192] In some implementations, R 2s is -L 2s -R 2a is. In some implementations, R s2 is R 2a is. In some implementations, R s2 is R. In some implementations, R 2s is -OL 2s' -R 2a and, here L 2s' is a covalent bond, or C 1-10 C having 1 to 4 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 It is a divalent, optionally substituted group selected from heteroaliphatic groups, where L 2s' One or more methylene units of are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)-, or -C(O)O-. In some embodiments, L 2s' is a covalent bond. In some embodiments, L 2s' is an optionally substituted 2-valent C 1-10 It is an aliphatic group. In some embodiments, L 2s' is 2 is C 1-10 It is an aliphatic group. In some embodiments, L 2s' is an optionally substituted divalent linear C 1-10It is an aliphatic group. In some embodiments, L 2s' is linear C 1-10 It is an aliphatic group. In some embodiments, L 2s' is optionally substituted C 1-10 It is an alkylene group. In some embodiments, L 2s' is C 1-10 It is an alkylene group. In some embodiments, L 2s' is an optionally substituted -(CH2) 1-10 -is. In some implementations, L 2s' -(CH2) 1-10 -is. In some implementations, L 2s' is an optionally substituted divalent C having 1 to 4 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 It is a heteroaliphatic group. In some embodiments, one or more methylene units are independently replaced as described herein. For example, in some embodiments, R 2s is -O(CH2) 0-10 R 2a and, where each -CH2- is optionally substituted. In some embodiments, R 2a is optionally substituted C 1-10 It is aliphatic. In some embodiments, R 2a is optionally substituted C 1-10 It is an alkyl. In some embodiments, R 2a is an optionally substituted linear C 1-10 It is an alkyl. In some embodiments, R 2a is C 1-10 It is an alkyl. In some embodiments, R 2a is C 1-10 It is a linear alkyl. In some embodiments, R 2s is -O(CH2) 0-15 It is CH3. In some implementations, R 2s is -O(CH2) 15 It is CH3.

[0193] In some implementations, R 2s is -L 2s -OR 2ais. In some implementations, R 2s is -OR 2a is. In some implementations, R 2s is -OMe.

[0194] In some implementations, R 2s is -L 2s -SR 2a is. In some implementations, R 2s is -SR 2a am.

[0195] In some implementations, R 2s is -L 2s -N(R 2a )2. In some implementations, R 2s is -N(R 2a )2 is.

[0196] In some implementations, R 2s is -OL 2s -OR 2a is. In some implementations, R 2s is -OL 2s -SR 2a is. In some implementations, R 2s is or -OL 2s -N(R 2a )2. In some embodiments, L 2s is an optionally substituted 2-valent C 2-10 It is an aliphatic group. In some embodiments, L 2s is 2 is C 2-10 It is an aliphatic group. In some embodiments, L 2s is linear C 2-10 It is an aliphatic group. In some embodiments, L 2s is linear C 2-10 It is an alkylene group. In some embodiments, R 2s is -OCH2CH2OCH3.

[0197] In some implementations, R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s and, here L sis L as described herein. For example, in some embodiments, L is (C2)-O-(unsubstituted methylene)-. In some embodiments, L is (C2)-O-(substituted methylene)-. In some embodiments, L is (C2)-O-(substituted methylene)-, where the methylene group is substituted with methyl. In some embodiments, L is (C2)-O-(substituted methylene)-, where the methylene group is substituted with ethyl. In some embodiments, the carbon atom of the methylene group is R is. In some embodiments, the carbon atom of the methylene group is S am.

[0198] In some embodiments, C2 and C4 are connected via -O-CH2-, where -CH2- is optionally substituted. In some embodiments, C2 and C4 are connected via -O-CH2-. In some embodiments, C2 and C4 are connected via -O-CH(CH3)-. In some embodiments, C2 and C4 are connected via -O-CH(CH2OMe)-. In some embodiments, L 2s The chiral carbon arrangement of R is. In some embodiments, L 2s My chiral carbon arrangement is S is. In some embodiments, -O- is bonded to C2. In some embodiments, the sugar is an LNA sugar.

[0199] L 2s

[0200] In some embodiments, L 2s is a covalent bond. In some embodiments, L 2s is an optionally substituted 2-valent C 1-10It is an aliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)-, or -C(O)O-. In some embodiments, L 2s is an optionally substituted 2-valent C 1-5 It is an aliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)-, or -C(O)O-. In some embodiments, L 2s is an optionally substituted 2-valent C 1-5 It is an aliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-. In some embodiments, L 2s is an optionally substituted 2-valent C 1-10 It is an aliphatic group. In some embodiments, L 2s is an optionally substituted 2-valent C 2-10 It is an aliphatic group. In some embodiments, L 2s is 2 is C 2-10 It is an aliphatic group. In some embodiments, L 2s is linear C 2-10 It is an aliphatic group. In some embodiments, L 2s is linear C 1-10 It is an alkylene group. In some embodiments, L2s is linear C 2-10 It is an alkylene group. In some embodiments, L 2s is an optionally substituted -O-CH2-.

[0201] R 2a

[0202] As defined herein, R 2a is R' as described herein. In some embodiments, R 2a is R as described herein. In some embodiments, R 2a is -H. In some implementations, R 2a is C 1-6 It is aliphatic. In some embodiments, R 2a is C 1-6 It is an alkyl. In some embodiments, R 2a is methyl.

[0203] In some embodiments, two or more R 2a is R', and together with the interposed atom(s), it forms an optionally substituted 3- to 20-membered ring having 0-5 heteroatoms independently selected from oxygen, nitrogen, and sulfur in addition to the interposed atom(s). In some embodiments, two R's attached to the same atom. 2a is R', and together with the atoms to which they are attached, they form an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20, 3 to 20, 5 to 9, etc.) ring having 0 to 5 heteroatoms independently selected from oxygen, nitrogen, and sulfur in addition to the interposed atoms. In some embodiments, the formed ring is as described herein.

[0204] L

[0205] Specific implementation examples for L are described below. Various variables, e.g., L 2s , L sThe back can be L, and an example of an implementation for L can also be applied to variables that can be L.

[0206] In some embodiments, L is a covalent bond. In some embodiments, L is a divalent optionally substituted C 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 e.g.) an aliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)-, or -C(O)O-. In some embodiments, L is a divalent optionally substituted C 1-6 It is an aliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)-, or -C(O)O-. In some embodiments, L is a divalent optionally substituted C 1-5It is an aliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)-, or -C(O)O-. In some embodiments, each methylene unit is replaced. In some embodiments, at least one methylene unit is not replaced. In some embodiments, L comprises at least one chain carbon atom. In some embodiments, L is -O-. In some embodiments, L is -S-. In some embodiments, L is -N(R')-. In some implementations, L is -C(O)-.

[0207] In some embodiments, no methylene unit is replaced. In some embodiments, L is a divalent optionally substituted C 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 et al.) It is an aliphatic group. In some embodiments, L is an optionally substituted C 1-6 It is an aliphatic group. In some embodiments, L is a divalent optionally substituted C 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 et al.) are alkylenes. In some embodiments, L is optionally substituted C 1-6 It is an alkylene. In some embodiments, L is linear. In some embodiments, L is branched. In some embodiments, L is substituted. In some embodiments, L is not substituted.

[0208] In some embodiments, L is a divalent, optionally substituted C having 1 to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Heteroaliphatic (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 (etc.) group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-. In some embodiments, L is a divalent, optionally substituted C having 1 to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-6 It is a heteroaliphatic group, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-. In some embodiments, L is linear. In some embodiments, L is branched. In some embodiments, L is substituted. In some embodiments, L is unsubstituted.

[0209] In some embodiments, the methylene unit is replaced with -Cy- as described herein.

[0210] -Cy-

[0211] As used herein, -Cy- is an optionally substituted divalent 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) circular ring having 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, -Cy- is an optionally substituted divalent 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) ring having 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, -Cy- is monocyclic. In some embodiments, -Cy- is dicyclic. In some embodiments, -Cy- is polycyclic. In some embodiments, each monocyclic ring unit is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 5, 1 to 4, 1 to 2, etc.) ring having 0 to 5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 to 5, 1 to 4, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, and 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 6, 7, 8, 9, 10, 3 to 8, 3 to 7, 3 to 6, 5 to 8, etc.) carbon atoms. In some embodiments, the monocyclic ring unit is an optionally substituted saturated ring. In some embodiments, the monocyclic ring unit is an optionally substituted partial unsaturated ring.In some embodiments, the monocyclic ring unit is an optionally substituted aromatic ring. In some embodiments, the monocyclic ring unit is an optionally substituted phenyl ring. In some embodiments, the monocyclic ring unit is an optionally substituted pentagonal heteroaromatic having 1 to 4 (e.g., 1, 2, 3, 4, 1 to 3, 1 to 2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the monocyclic ring unit is an optionally substituted hexagonal heteroaromatic having 1 to 4 (e.g., 1, 2, 3, 4, 1 to 3, 1 to 2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur. Each monocyclic ring unit is independently optionally substituted. In some embodiments, the monocyclic ring unit is unsaturated. In some embodiments, the monocyclic ring unit is saturated. In some embodiments, -Cy- is an optionally substituted saturated ring. In some embodiments, -Cy- is an optionally substituted partial unsaturated ring. In some embodiments, -Cy- is an optionally substituted aromatic ring.

[0212] In some embodiments, -Cy- is an optionally substituted divalent 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) monocyclic alicyclic ring. In some embodiments, -Cy- is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) monocyclic cycloalkyl ring. In some embodiments, -Cy- is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) monocyclic heteroaliphatic ring having 1 to 5 heteroatoms. In some embodiments, -Cy- is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, or 10) monocyclic heteroalkyl ring having 1 to 5 heteroatoms. In some embodiments, -Cy- is an optionally substituted divalent 5 to 15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) monocyclic or polycyclic alicyclic group. In some embodiments, -Cy- is an optionally substituted divalent 5 to 15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15) monocyclic or polycyclic cycloalkyl group. In some embodiments, -Cy- is an optionally substituted 5 to 15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15)-membered cyclic or polycyclic heteroaliphatic ring having 1 to 5 heteroatoms. In some embodiments, -Cy- is an optionally substituted 5 to 15 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15)-membered cyclic or polycyclic heterocyclyl ring having 1 to 5 heteroatoms. In some embodiments, the alicyclic, cycloalkyl, heteroaliphatic, or heteroalkyl ring is ternary. In some embodiments, it is quaternary. In some embodiments, it is quinary. In some embodiments, it is six-membered. In some embodiments, this is 7 won. In some embodiments, this is 8 won. In some embodiments, this is 9 won. In some embodiments, this is 10 won.In some embodiments, this is 11 members. In some embodiments, this is 12 members. In some embodiments, -Cy- is optionally substituted phenylene. In some embodiments, -Cy- is an optionally substituted divalent 10-membered aryl ring. In some embodiments, -Cy- is an optionally substituted 5-membered heteroaryl ring having 1 to 4 heteroatoms. In some embodiments, -Cy- is an optionally substituted 6-membered heteroaryl ring having 1 to 4 heteroatoms. In some embodiments, -Cy- is an optionally substituted 9-membered heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, -Cy- is an optionally substituted 10-membered heteroaryl ring having 1 to 5 heteroatoms. In some embodiments, the heteroaliphatic, heterocyclil, or heteroaryl ring contains one or fewer heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur.

[0213] In some embodiments, -Cy- is an optionally substituted 4- to 7-membered ring having 0 to 3 heteroatoms. In some embodiments, -Cy- is an optionally substituted 6-membered aryl ring. In some embodiments, the aryl ring is substituted. In some embodiments, it is substituted with one or more halogens. In some embodiments, it is substituted with one or more -F. In some embodiments, it is not substituted. In some embodiments, it is optionally substituted is. In some implementations, this is. In some embodiments, this is optionally substituted is. In some implementations, this is. In some embodiments, this is optionally substituted is. In some implementations, this is. In some embodiments, -Cy- is an optionally substituted pentagonal heteroaryl ring having 1 to 3 heteroatoms. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. In some embodiments, -Cy- is optionally substituted is. In some implementations, -Cy- is am.

[0214] R 1

[0215] In some implementations, R 1 is R' as described herein and is not -H. In some embodiments, R 1 is R as described herein. In some embodiments, R 1 is optionally substituted C 1-6 It is aliphatic. In some embodiments, R 1 is an optionally substituted methyl. In some embodiments, R 1 is an optionally substituted ethyl. In some embodiments, R 1 is an optionally substituted ethyl, wherein the methylene unit bonded to the oxygen is optionally single-substituted, and the methyl is substituted with an electron-withdrawing group, e.g., -CN. Specific electron-withdrawing groups are described in US 10167309, US 11643657, US 11718638, US 11608355, or US 20230089442, each of which is incorporated herein by reference. In some embodiments, R 1 is -OCH2CH2CN.

[0216] R 2

[0217] In some implementations, R 2 is R' as described herein and is not -H. In some embodiments, R 2 is R as described herein. In some embodiments, R 2is not -H. In some implementations, R 2 is optionally substituted C 1-10 It is aliphatic. In some embodiments, R 2 is optionally substituted C 1-10 It is an alkyl. In some embodiments, R 2 C 1-6 It is aliphatic. In some embodiments, R 2 C 1-6 It is an alkyl. In some embodiments, R 2 It is isopropyl.

[0218] R 3

[0219] In some implementations, R 3 is R' as described herein and is not -H. In some embodiments, R 3 is R as described herein. In some embodiments, R 3 is not -H. In some implementations, R 3 is optionally substituted C 1-10 It is aliphatic. In some embodiments, R 3 is optionally substituted C 1-10 It is an alkyl. In some embodiments, R 2 C 1-6 It is aliphatic. In some embodiments, R 2 C 1-6 It is an alkyl. In some embodiments, R 3 It is isopropyl.

[0220] In some implementations, R 2 and R 3 They are identical. In some implementations, they are different.

[0221] In some implementations, -P(OR 1 )N(R 2 )(R 3 ) is -P(OCH2CH2CN)N[CH(CH3)2]2.

[0222] In some implementations, R 1 , R 2 and R 3Take two or three of them together with the atoms interposed between them It forms, wherein each variable is independently as described herein. In some embodiments, R 1 and R 2 is taken together with an atom interposed therein to form a ring as described herein. In some embodiments, R 2 and R 3 It is taken together with an atom interposed therebetween to form a ring as described herein. In some embodiments, R 1 , R 2 and R 3 It is taken together with an atom interposed therein to form a ring as described herein.

[0223] Ring A

[0224] As defined herein, ring A is an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 3, 4, 5, 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) ring having 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 16, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur in addition to the interposed atoms. In some embodiments, ring A is an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) circular ring having 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, ring A does not have additional heteroatoms. In some embodiments, the ring has 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional heteroatoms.

[0225] In some embodiments, ring A is not substituted (as will be known to those skilled in the art, R s (excluding ). In some embodiments, ring A is substituted (as will be known to those skilled in the art, R s (Does not include ).

[0226] In some embodiments, ring A is monocyclic. In some embodiments, ring A is dicyclic. In some embodiments, ring A is polycyclic. In some embodiments, each monocyclic ring unit is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 4, 5, 1 to 5, 1 to 4, 1 to 2, etc.) ring having 0 to 5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 to 5, 1 to 4, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, and 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 3 to 8, 3 to 7, 3 to 6, 4 to 6, 5 to 6, 5 to 8, etc.) carbon atoms. In some embodiments, each monocyclic ring unit is an optionally substituted 4 to 10 (e.g., 4, 5, 6, 7, 8, 9, 10, 5 to 7, 5 to 6)-membered ring having 0 to 5 (e.g., 1, 2, 3, 4, 5, 1 to 5, 1 to 4, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, and 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 to 5, 1 to 4, etc.) carbon atoms. In some embodiments, each monocyclic ring unit is an optionally substituted 3 to 10-membered ring. In some embodiments, each monocyclic ring unit is an optionally substituted 3 to 7-membered ring. In some embodiments, each monocyclic ring unit is independently and optionally substituted with 4 to 6 members. In some embodiments, each monocyclic ring unit is independently and optionally substituted with 5 to 6 members. In some embodiments, each monocyclic ring unit is independently and optionally substituted with 5 members. In some embodiments, the monocyclic ring unit is an optionally substituted saturated ring. In some embodiments, the monocyclic ring unit is an optionally substituted partial unsaturated ring. In some embodiments, the monocyclic ring unit is an optionally substituted aromatic ring. In some embodiments, the monocyclic ring unit is an optionally substituted phenyl ring.In some embodiments, the single ring unit is an optionally substituted pentagonal heteroaromatic having 1 to 4 heteroatoms (e.g., 1, 2, 3, 4, 1 to 3, 1 to 2, etc.) independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the single ring unit is an optionally substituted hexagonal heteroaromatic having 1 to 4 heteroatoms (e.g., 1, 2, 3, 4, 1 to 3, 1 to 2, etc.) independently selected from nitrogen, oxygen, and sulfur. Each single ring unit is independently optionally substituted. In some embodiments, the single ring unit is unsaturated. In some embodiments, the single ring unit is saturated.

[0227] In some implementations, R 2 and R 3 The monocyclic ring unit containing this bonded nitrogen atom is an optionally substituted saturated 4–10 (e.g., 4–8, 4–6, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0–2 heteroatoms independently selected from nitrogen, oxygen, and sulfur in addition to the nitrogen atom. In some embodiments, R 2 and R 3 The single ring unit containing this bonded nitrogen atom is an optionally substituted saturated 4 to 10 (e.g., 4 to 8, 4 to 6, 4, 5, 6, 7, 8, 9, 10, etc.) ring that does not have a heteroatom independently selected from nitrogen, oxygen, and sulfur in addition to the nitrogen atom. In some embodiments, it is 4-membered. In some embodiments, it is 5-membered. In some embodiments, it is 6-membered. In some embodiments, it is 7-membered. In some embodiments, it is 8-membered.

[0228] In some embodiments, ring A is an optionally substituted saturated ring. In some embodiments, ring A is an optionally substituted partially unsaturated ring. In some embodiments, ring A is an optionally substituted aromatic ring.

[0229] In some embodiments, ring A is an optionally substituted monocyclic 3 to 10 (e.g., 3 to 6, 5 to 10, 5 to 6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur in addition to the interposed atom(s). In some embodiments, ring A is an optionally substituted dicyclic or polycyclic ring of 6 to 20 (e.g., 6 to 15, 6 to 10, 8 to 20, 8 to 15, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur in addition to the interposed atom(s). In some embodiments, ring A is 3 to 9, 3 to 8, 3 to 7, 3 to 6, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10 won. In some embodiments, ring A is 3 to 9 won. In some embodiments, ring A is 3 to 7 won. In some embodiments, ring A is 4 to 10 won. In some embodiments, ring A is 4 to 7 won. In some embodiments, ring A is 5 to 10 won. In some embodiments, ring A is 5 to 7 won. In some embodiments, ring A is 3 won. In some embodiments, ring A is 4 won. In some embodiments, ring A is 5 won. In some embodiments, ring A is 6 units. In some embodiments, ring A is 7 units. In some embodiments, ring A is 8 units. In some embodiments, ring A is 9 units. In some embodiments, ring A is 10 units. In some embodiments, ring A is 11 units. In some embodiments, ring A is 12 units. In some embodiments, ring A is monocyclic. In some embodiments, ring A is bicyclic. In some embodiments, ring A is optionally substituted bicyclic rings of 7 to 12 units. In some embodiments, ring A is optionally substituted bicyclic rings of 8 to 10 units.In some embodiments, ring A is an optionally substituted cyclic hepta-ring. In some embodiments, ring A is an optionally substituted cyclic octa-ring. In some embodiments, ring A is an optionally substituted cyclic ninta-ring. In some embodiments, ring A is an optionally substituted cyclic tenta-ring. In some embodiments, ring A is polycyclic. In some embodiments, ring A does not have additional heteroatoms. In some embodiments, ring A has 0 to 6 additional heteroatoms, e.g., 0, 1 to 6, 1 to 5, 1 to 3, or 1, 2, 3, 4, 5, or 6. In some embodiments, ring A includes one or more aromatic rings. In some embodiments, ring A is dicyclic or polycyclic, and each monocyclic ring unit is independently 3 to 10 (e.g., 3 to 9, 3 to 8, 3 to 7, 3 to 6, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10) atoms, saturated, partially unsaturated, or aromatic, and has 0 to 5 (e.g., 0, 1 to 5, 1 to 3, 1, 2, 3, 4, or 5) heteroatoms. In some embodiments, the monocyclic ring unit is saturated. In some embodiments, the monocyclic ring unit is partially unsaturated. In some embodiments, the monocyclic ring unit is aromatic. In some embodiments, the monocyclic ring unit is heteroaromatic. A person skilled in the art recognizes that, for example, the intervening atom(s) of a group taken together to form a ring are generally atoms in the shortest path connecting these groups where multiple paths exist.

[0230] In some embodiments, the single-ring unit containing phosphorus, nitrogen, and oxygen atoms is saturated. In some embodiments, the single-ring unit containing phosphorus, nitrogen, and oxygen atoms is partially saturated. In some embodiments, it is quaternary. In some embodiments, it is quintary. In some embodiments, it is sixtary. In some embodiments, it is seventary. In some embodiments, it has one or more additional heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, phosphorus, nitrogen, and oxygen atoms are the only heteroatoms of the corresponding single-ring unit.

[0231] In some implementations, R 2 and R 3 The monocyclic ring unit containing this bonded nitrogen atom is an optionally substituted partially unsaturated 4–10 (e.g., 4–8, 4–6, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0–2 heteroatoms independently selected from nitrogen, oxygen, and sulfur in addition to the nitrogen atom. In some embodiments, R 2 and R 3 The monocyclic ring unit containing this bonded nitrogen atom is an optionally substituted partially unsaturated 4 to 10 (e.g., 4 to 8, 4 to 6, 4, 5, 6, 7, 8, 9, 10, etc.) member ring that does not have a heteroatom independently selected from nitrogen, oxygen, and sulfur in addition to the nitrogen atom. In some embodiments, it is 4. In some embodiments, it is 5. In some embodiments, it is 6. In some embodiments, it is 7. In some embodiments, it is 8.

[0232] In some embodiments, ring A is optionally substituted Includes, where each L a is independently a covalent bond or divalent C 1-5 It is an aliphatic group, and each L aOne or more methylene units of are optionally and independently replaced with -O-, -S-, or -NH. In some embodiments, ring A is optionally substituted and, here, each L a is independently a covalent bond or divalent C 1-5 It is an aliphatic group, and each L a One or more methylene units of are optionally and independently replaced with -O-, -S-, or -NH. In some embodiments, ring A is optionally substituted Includes, where each L a is independently a covalent bond or divalent C 1-5 It is an aliphatic group, and each L a One or more methylene units of are optionally and independently replaced with -O-, -S-, or -NH. In some embodiments, ring A is optionally substituted and, here, each L a is independently a covalent bond or divalent C 1-5 It is an aliphatic group, and each L a One or more methylene units of are optionally and independently replaced with -O-, -S-, or -NH. In some embodiments, L a is a covalent bond. In some embodiments, L a is not a covalent bond. In some embodiments, L a is -CH2-. In some implementations, L a is -(CH2)2-. In some embodiments, L a is -(CH2)3-. In some embodiments, L a is -(CH2)4-. In some embodiments, L a is -(CH2)5-. In some embodiments, the methylene unit is replaced with -O-. In some embodiments, the methylene unit is replaced with -S-. In some embodiments, the methylene unit is replaced with -NH-.

[0233] In some embodiments, ring A is optionally substituted is. In some embodiments, ring A is optionally substituted is. In some embodiments, ring A is optionally substituted am.

[0234] In some implementations, R s The existence of -P(OR 1 )N(R 2 )(R 3 It is bonded to the carbon atom bonded to the oxygen of ). In some embodiments, -P(OR 1 )N(R 2 )(R 3 )Is and, here R s1 and R s2 Each is independently R as described herein s is. In some implementations, -P(OR 1 )N(R 2 )(R 3 )silver is. In some implementations, -P(OR 1 )N(R 2 )(R 3 )silver is. In some implementations, -P(OR 1 )N(R 2 )(R 3 )silver is. In some implementations, -P(OR 1 )N(R 2 )(R 3 )silver is. In some implementations, -P(OR 1 )N(R 2 )(R 3 )silver am.

[0235] In some implementations, R s1 and R s2 One of them is -H. In some implementations, R s1 and R s2 is not independently -H. In some implementations, R s1 is R as described herein. In some embodiments, R s1is not hydrogen. In some embodiments, R s1 is hydrogen. In some embodiments, R s1 is hydrogen and R s2 is not hydrogen. In some embodiments, R s1 is optionally substituted C 1-6 It is aliphatic. In some embodiments, R s1 is methyl. In some embodiments, R s1 is an optionally substituted phenyl. In some embodiments, R s1 is phenyl. In some embodiments, R s2 is R as described herein. In some embodiments, R s2 is not hydrogen. In some embodiments, R s2 is hydrogen. In some embodiments, R s2 is optionally substituted C 1-6 It is aliphatic. In some embodiments, R s2 is methyl. In some embodiments, R s2 is an optionally substituted phenyl. In some embodiments, R s2 is phenyl.

[0236] In some implementations, -P(OR 1 )N(R 2 )(R 3 )Is and, each variable is as independently described herein. In some embodiments, Is a has a structure that is a chiral auxiliary chain as described, for example, in US 10167309, US 11643657, US 11718638, US 11608355 or US 20230089442, each of which is incorporated herein by reference. For example, in some embodiments, it is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this is. In some implementations, this am.

[0237] t

[0238] In some embodiments, t is 0. In some embodiments, t is 1 to 5. In some embodiments, t is 1. In some embodiments, t is 2. In some embodiments, t is 3. In some embodiments, t is 4. In some embodiments, t is 5.

[0239] R s

[0240] In some implementations, R s is -F. In some implementations, R s is -Cl. In some implementations, R s is -Br. In some cases, R s is -I. In some cases, R s is -CN. In some implementations, R s is -N3. In some implementations, R s is -NO. In some implementations, R s is -NO2.

[0241] In some implementations, R s is -L s -R s11 and, here L s and R s11 Each is as independently described herein. In some embodiments, R s R as described hereins11 am.

[0242] In some implementations, R s is R' as described herein. For example, in some embodiments, R' is -C(O)R, where R is as described herein. In some embodiments, R' is -CO2R, where R is as described herein. In some embodiments, R' is -S(O)2R, where R is as described herein. In some embodiments, R is C 1-6 It is aliphatic. In some embodiments, R is C 1-6 It is an alkyl. In some embodiments, R is optionally a substituted phenyl. In some embodiments, R s is -S(O)2R, where R is an optionally substituted C 1-6 It is aliphatic. In some embodiments, R s is -S(O)2Me. In some implementations, R s is -S(O)2t-Bu. In some implementations, R s is -S(O)2R, where R is an optionally substituted phenyl. In some embodiments, R s is -S(O)2Ph.

[0243] In some implementations, R s is R as described herein. For example, in some embodiments, R is not -H. In some embodiments, R is optionally a substituted C 1-6 It is aliphatic. In some embodiments, R is optionally substituted C 1-6 It is an alkyl. In some embodiments, R is methyl. In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl.

[0244] In some implementations, R s is -L s -OR s11 and, here L s and R s11 Each is as independently described herein. In some embodiments, R sis -OR s11 and, here R s11 ... is as described herein. In some embodiments, R s -CH2OR s11 and, here R s11 It is as described in the present invention.

[0245] In some implementations, R s is -L s -SR s11 and, here L s and R s11 Each is as independently described herein. In some embodiments, R s is -SR s11 and, here R s11 ... is as described herein. In some embodiments, R s -CH2SR s11 and, here R s11 It is as described in the present invention.

[0246] In some implementations, R s is -L s -N(R s11 )2 and, here L s and R s11 Each is as independently described herein. In some embodiments, R s is -N(R s11 )2 and, here L s and R s11 Each is as independently described herein. In some embodiments, R s is -CH2N(R s11 )2 and, here L s and R s11 Each is as described independently in the present invention.

[0247] In some implementations, R s is -C(R s11 )3 and here, each R s11 ...is as independently described herein. In some embodiments, R s is -CH(R s11 )2 and here, each Rs11 ...is as independently described herein. In some embodiments, two or more R s11 and the carbon atoms to which they are attached are taken together to form an optionally substituted 3 to 20 (e.g., 3 to 15, 3 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0 to 10 heteroatoms (e.g., 0, 1 to 10, 1 to 5, 1 to 4, 1 to 6, 5 to 10, 5 to 6, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, R s is optionally substituted am.

[0248] In some implementations, R s is -L s -Si(R s11 )3 and here, each R s11 ...is as independently described herein. In some embodiments, each R s11 is independently R' as described herein and is not -H. In some embodiments, each R s11 is independently R as described herein and is not -H. In some embodiments, at least one R s11 is optionally substituted C 1-10 It is aliphatic. In some embodiments, at least one R s11 is optionally substituted C 1-10 It is an alkyl. In some embodiments, at least one R s11 is methyl. In some embodiments, at least one R s11 is an optionally substituted phenyl. In some embodiments, at least one R s11 is phenyl. In some embodiments, each R s11 C independently 1-10 It is an optionally substituted group selected from aliphatic and 6- to 10-membered aryls. In some embodiments, each Rs11 C independently 1-10 It is an optionally substituted group selected from alkyl and 6- to 10-membered aryls. In some embodiments, each R s11 C independently 1-6 It is an optionally substituted group selected from alkyl and 6- to 10-phenyl. In some embodiments, R s11 It is -SiPh2Me.

[0249] R s11

[0250] As used herein, R s11 is R' as described herein. For example, in some embodiments, R' is R as described herein. In some embodiments, R' is -C(O)R, where R is as described herein. In some embodiments, R' is -CO2R, where R is as described herein. In some embodiments, R' is -S(O)2R, where R is as described herein. R s11 is -H. In some implementations, R s11 It is not -H. In some embodiments, two or more Rs on the same atom s11 The atom(s) interposed therebetween are taken together to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s). Specific embodiments of the ring are described herein.

[0251] L s

[0252] As used herein, L s is L as described herein. For example, in some embodiments, L is a covalent bond. In some embodiments, L is optionally a substituted -CH2-. In some embodiments, L is -CH2-. In some embodiments, L is a singly substituted -CH2-.

[0253] R’

[0254] A specific implementation example for R' is described below. Various variables, for example, R 2a , R 1 , R 2 , R 3 , R s , R s11 The etc. can be R', and an example of an implementation for R' can also be applied to these variables that can be R'.

[0255] In some embodiments, R' is R as described herein. In some embodiments, R' is -H. In some embodiments, R' is not -H.

[0256] In some embodiments, R' is -C(O)R, where R is as described herein. In some embodiments, R' is -CO2R, where R is as described herein. In some embodiments, R' is -S(O)2R, where R is as described herein. In some embodiments, as described herein, R is not -H.

[0257] In some embodiments, two or more R's and an atom(s) interposed therebetween are taken together to form an optionally substituted 3 to 20 (e.g., 3 to 15, 3 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0 to 10 heteroatoms (e.g., 0, 1 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s). In some embodiments, two or more R's on the same atom and the atoms to which they are attached are taken together to form an optionally substituted 3 to 20 (e.g., 3 to 15, 3 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0 to 10 heteroatoms (e.g., 0, 1 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1 to 3, 4, 5, 6, 7, 8, 9, 10, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms. In some embodiments, two R's on the same atom and the atoms to which they are attached are taken together to form an optionally substituted 3 to 20 (e.g., 3 to 15, 3 to 10, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) ring having 0 to 10 heteroatoms (e.g., 0, 1 to 10, 1 to 5, 1 to 4, 1 to 3, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms.

[0258] In some embodiments, each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's on the same atom and the atoms to which they are attached are taken together to form an optionally substituted 3 to 20-membered ring having 0 to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s).

[0259] ring

[0260] The compounds of the present disclosure may contain various rings. In some embodiments, a variable, which may be, for example, R or R', may be a ring as described herein. In some embodiments, two variables, for example, two R' groups, may be taken together with interposed atom(s) to form a ring as described herein. In some embodiments, the ring is monovalent. In some embodiments, the ring is divalent. In some embodiments, the ring structure may be polyvalent. The ring is optionally substituted. In some embodiments, the ring is unsubstituted. In some embodiments, the ring is substituted.

[0261] Specific embodiments and features of the ring are described below as examples.

[0262] In some embodiments, the ring is an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) circular ring having 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the ring is an optionally substituted 3 to 20 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 10, 0 to 5, 1 to 5, etc.) circular ring having 0 to 10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3 to 15, 3 to 14, 3 to 10, 5 to 10, 3 to 7, 3 to 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, the ring is a monocyclic ring. In some embodiments, the ring is a dicyclic ring. In some embodiments, the ring is a polycyclic ring.

[0263] In some embodiments, each monocyclic ring unit is an optionally substituted 3 to 10 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1 to 5, 1 to 4, 1 to 2, etc.) ring having 0 to 5 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 to 5, 1 to 4, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, the monocyclic ring unit is an optionally substituted saturated ring. In some embodiments, the monocyclic ring unit is an optionally substituted partially unsaturated ring. In some embodiments, the monocyclic ring unit is an optionally substituted aromatic ring. In some embodiments, the monocyclic ring unit is an optionally substituted phenyl ring. In some embodiments, the monocyclic ring unit is an optionally substituted pentagonal heteroaromatic having 1 to 4 (e.g., 1, 2, 3, 4, 1 to 3, 1 to 2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the monocyclic ring unit is an optionally substituted hexagonal heteroaromatic having 1 to 4 (e.g., 1, 2, 3, 4, 1 to 3, 1 to 2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur. Each monocyclic ring unit is independently optionally substituted. In some embodiments, the monocyclic ring unit is unsaturated. In some embodiments, the monocyclic ring unit is saturated.

[0264] In some embodiments, the ring is an optionally substituted saturated ring. In some embodiments, the ring is an optionally substituted partially unsaturated ring. In some embodiments, the ring is an optionally substituted aromatic ring.

[0265] In some embodiments, the ring is an optionally substituted 3 to 10-membered monocyclic, bicyclic, or polycyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur in addition to the interposed atom(s). In some embodiments, the ring is 3 to 9, 3 to 8, 3 to 7, 3 to 6, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9, or 10 members. In some embodiments, the ring is 3 to 9 members. In some embodiments, the ring is 3 to 7 members. In some embodiments, the ring is 4 to 10 members. In some embodiments, the ring is 4 to 7 members. In some embodiments, the ring is 5 to 10 members. In some embodiments, the ring is 5 to 7 units. In some embodiments, the ring is 3 units. In some embodiments, the ring is 4 units. In some embodiments, the ring is 5 units. In some embodiments, the ring is 6 units. In some embodiments, the ring is 7 units. In some embodiments, the ring is 8 units. In some embodiments, the ring is 9 units. In some embodiments, the ring is 10 units. In some embodiments, the ring is a monoring. In some embodiments, the ring is a biring. In some embodiments, the ring is a polyring. In some embodiments, the ring does not have heteroatoms. In some embodiments, the ring has 1 to 6, e.g., 1 to 5, 1 to 3, or 1, 2, 3, 4, 5, or 6 heteroatoms. In some embodiments, a ring formed by taking two or more groups and interposed atoms(s) together does not have heteroatoms other than the interposed atoms(s). In some embodiments, a ring formed by taking two or more groups and interposed atoms(s) together has 1 to 6 heteroatoms in addition to the interposed atoms(s), e.g., 1 to 5, 1 to 3, or 1, 2, 3, 4, 5, or 6 heteroatoms. In some embodiments, the ring is saturated. In some embodiments, the ring is partially unsaturated. In some embodiments, the ring comprises one or more aromatic rings.In some embodiments, the ring is dicyclic or polycyclic, and each monocyclic ring unit is independently 3 to 10 (e.g., 3 to 9, 3 to 8, 3 to 7, 3 to 6, 4 to 10, 4 to 9, 4 to 8, 4 to 7, 4 to 6, 5 to 10, 5 to 9, 5 to 8, 5 to 7, 5 to 6, or 3, 4, 5, 6, 7, 8, 9 or 10) saturates, partially unsaturated, or aromatic, and has 0 to 5 (e.g., 0, 1 to 5, 1 to 3, 1, 2, 3, 4 or 5) heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the monocyclic ring unit is saturated. In some embodiments, the monocyclic ring unit is partially unsaturated. In some embodiments, the monocyclic ring unit is aromatic. In some embodiments, the monocyclic ring unit is heteroaromatic. Those skilled in the art recognize that, for example, the intervening atom(s) of the groups taken together to form a ring are generally atoms in the shortest path connecting these groups where multiple paths exist.

[0266] In some embodiments, the ring has one or more heteroatoms. In some embodiments, the ring includes a nitrogen atom. In some embodiments, the ring includes an oxygen atom. In some embodiments, the ring includes a sulfur atom.

[0267] R

[0268] Various variables of the present disclosure may independently be R. Specific embodiments of R are described below by way of example. Those skilled in the art reading the present disclosure recognize that embodiments described for a variable that may be R and falls within the definition of R may be embodiments of R. Those skilled in the art reading the present disclosure further recognize that embodiments of R, such as those described for R, those described for a variable that may be R and falls within the definition of R, etc., may be embodiments of a variable that may be R.

[0269] In some embodiments, R is -H. In some embodiments, R is not -H. In some embodiments, R is optionally a substituted C 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 et al.) are aliphatic. In some embodiments, R is optionally substituted C 1-8 It is aliphatic. In some embodiments, R is optionally substituted C 1-6 It is aliphatic. In some embodiments, R is optionally substituted C 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 (etc.) is an alkyl. In some embodiments, R is optionally a substituted C 1-8 It is an alkyl. In some embodiments, R is optionally a substituted C 1-6 It is aliphatic. In some embodiments, R is optionally substituted C 1-6 It is an alkyl. In some embodiments, R is an optionally substituted -CH2-C 3-6 It is a cycloaliphatic group. In some embodiments, R is an optionally substituted -CH2-C 3-6 It is a cycloaliphatic group. In some embodiments, R is an optionally substituted -CH2-C 3-6 It is cycloalkyl. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R is isopropyl. In some embodiments, R is -CF3. In some embodiments, R is -CH2CF3. In some embodiments, R is butyl. In some embodiments, R is t-butyl.

[0270] In some embodiments, R is an optionally substituted 3–10 (e.g., 3–9, 3–8, 3–7, 3–6, 5–7, 4, 5, 6, 7, 8, 9, 10, etc.) cycloaliphatic group. In some embodiments, R is an optionally substituted C 3-10It is cycloalkyl. In some embodiments, R is optionally substituted cyclopropyl. In some embodiments, R is optionally substituted cyclobutyl. In some embodiments, R is optionally substituted cyclopentyl. In some embodiments, R is optionally substituted cyclohexyl.

[0271] In some embodiments, R is an optionally substituted C having 1 to 5 heteroatoms (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 (etc.) is heteroaliphatic. In some embodiments, R is an optionally substituted C having 1 to 5 heteroatoms (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-6 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 e.g.) is heteroaliphatic. In some embodiments, R is an optionally substituted C having 1 to 5 heteroatoms (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) independently selected from oxygen, nitrogen, and sulfur. 1-10 (e.g., C 1-6 , C 2-8 , C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 e.g.) is heteroaliphatic. In some embodiments, R is an optionally substituted C having 1 to 5 heteroatoms (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) independently selected from oxygen, nitrogen, and sulfur. 1-6 (e.g., C 1-6 , C 2-8, C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 e.g.) It is heteroaliphatic. In some embodiments, R is an optionally substituted C1-6 heteroaliphatic having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted C1-6 heteroaliphatic having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, or a C1-6 heteroaliphatic having 1 heteroatom independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is a heteroalkyl.

[0272] In some embodiments, R is an optionally substituted 3 to 15 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 3 to 10, 3 to 7, 4 to 10, 4 to 6, etc.) heterocyclic ring having 1 to 5 (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) heterocyclic rings independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, R is an optionally substituted 3 to 15 (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 3 to 10, 3 to 7, 4 to 10, 4 to 6, etc.) heterocyclic ring having 1 to 5 (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, R is an optionally substituted 3 to 10 (e.g., 3 to 9, 3 to 8, 3 to 7, 3 to 6, 5 to 7, 4, 5, 6, 7, 8, 9, 10, etc.) heterocyclil having 1 to 4 (e.g., 1, 2, 3, 4, 1 to 2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted ternary heterocyclil having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted ternary heterocyclil having 1 heteroatom independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted quaternary heterocyclil having 1 to 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted pentagonal heterocyclil having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted hexagonal heterocyclil having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.In some embodiments, R is an optionally substituted hepta-heterocyclil having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted octa-heterocyclil having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted ko-heterocyclil having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted ten-heterocyclil having 1 to 3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, one carbon atom is present in the heterocyclil ring. In some embodiments, the heterocyclyl ring has two or more carbon atoms (e.g., 2 to 14, 2 to 9, 2 to 6, 2 to 4, 3 to 10, 3 to 5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, etc.).

[0273] In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl. In some embodiments, R is optionally substituted naphthyl. In some embodiments, R is optionally substituted 1-naphthyl. In some embodiments, R is optionally substituted 2-naphthyl. In some embodiments, R is naphthyl.

[0274] In some embodiments, R is an optionally substituted 5 to 14 (e.g., 5, 6, 9, 10, 14, etc.) heteroaryl having 1 to 6 (e.g., 1 to 5, 1 to 4, 1, 2, 3, 4, 5, 6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, R is an optionally substituted 5 to 14 (e.g., 5, 6, 9, 10, 14, etc.) heteroaryl having 1 to 6 (e.g., 1 to 5, 1 to 4, 1, 2, 3, 4, 5, 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, R is an optionally substituted 5 to 10 (e.g., 5, 6, 9, 10, etc.) heteroaryl having 1 to 6 (e.g., 1 to 5, 1 to 4, 1, 2, 3, 4, 5, 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. In some embodiments, R is an optionally substituted 5 to 10 heteroaryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the heteroaryl ring is monocyclic and 5-membered. In some embodiments, the heteroaryl ring is monocyclic and 6-membered. In some embodiments, the heteroaryl ring is bicyclic and 9-membered. In some embodiments, the heteroaryl ring is bicyclic and 10-membered. In some embodiments, the heteroaryl ring is tricyclic and 14-membered. In some embodiments, R is an optionally substituted pentagonal heteroaryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted pentagonal heteroaryl having 1 heteroatom selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted hexaagonal heteroaryl having 1 to 4 heteroatoms independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted hexaagonal heteroaryl having 1 heteroatom selected from nitrogen, oxygen, and sulfur.In some embodiments, R is an optionally substituted cyclic 8-10-membered aromatic ring having 1 to 6 heteroatoms of nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted cyclic 9-membered aromatic ring having 1 to 6 heteroatoms of nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted cyclic 10-membered aromatic ring having 1 to 6 heteroatoms of nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted cyclic 9-membered aromatic ring having 1 heteroatom selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an optionally substituted cyclic 10-membered aromatic ring having 1 heteroatom of nitrogen, oxygen, and sulfur. In some embodiments, at least one heteroatom is nitrogen. In some embodiments, at least one heteroatom is oxygen. In some embodiments, at least one heteroatom is sulfur. In some embodiments, each heteroatom is identical. In some embodiments, at least one heteroatom is different from the other heteroatoms.

[0275] As described herein, various groups may be optionally substituted. Substituents are routinely used in chemistry, including in the development of various therapeutic agents. Various substituents may be utilized according to the present disclosure. In some embodiments, the optionally substituted group is not substituted. In some embodiments, the optionally substituted group is substituted. The substituents preferably result in the formation of a compound for desired properties, activity, use, etc., as described herein. In some embodiments, the compound is stable for therapeutic use as described herein. The term "stable" refers, as used herein, to a compound that is substantially unchanged when subjected to conditions that allow for the production, detection, and, in certain embodiments, recovery, purification, and use for one or more purposes disclosed herein. In some embodiments, the substituent is a hydrocarbon group. In some embodiments, the substituent comprises a heteroatom. In some embodiments, the substituent comprises a plurality of heteroatoms. In some embodiments, each atom of the substituent is independently selected from hydrogen, carbon, halogen, nitrogen, oxygen, sulfur, phosphorus, and silicon. In some embodiments, each atom of the substituent is independently selected from hydrogen, carbon, halogen, nitrogen, oxygen, and sulfur. In some embodiments, each atom of the substituent is independently selected from hydrogen, carbon, fluorine, chlorine, bromine, iodine, nitrogen, oxygen, and sulfur.In some embodiments, the total number of carbon and non-halogen heteroatom(s) of the substituent is about 1 or about 1 or less; in some embodiments, it is about 2 or less; in some embodiments, it is about 3 or less; in some embodiments, it is about 4 or less; in some embodiments, it is about 5 or less; in some embodiments, it is about 6 or less; in some embodiments, it is about 7 or less; in some embodiments, it is about 8 or less; in some embodiments, it is about 9 or less; in some embodiments, it is about 10 or less; in some embodiments, it is about 11 or less; in some embodiments, it is about 12 or less; in some embodiments, it is about 13 or less; in some embodiments, it is about 14 or less; in some embodiments, it is about 15 or less; in some embodiments, it is about 20 or less. In some embodiments, the total number of carbon and non-halogen heteroatom(s) of each substituent is independently about 20 or fewer. In some embodiments, the total number of carbon and non-halogen heteroatom(s) of each substituent is independently about 15 or fewer. In some embodiments, the total number of carbon and non-halogen heteroatom(s) of each substituent is independently about 10 or fewer. In some embodiments, the total number of carbon and non-halogen heteroatom(s) of each substituent is independently about 6 or fewer.

[0276] nucleoside

[0277] Various nucleosides, e.g., those described in US 10167309, US 11643657, US 11718638, US 11608355, US 20230089442, etc., may be utilized for the production of oligonucleotides. In some embodiments, the present disclosure is as -P(O)(OR) as described herein. PG The present invention provides a nucleoside comprising )2. In some embodiments, the nucleoside is -P(O)(OR )2 coupled to the sugar moiety of the nucleoside. PGIncludes )2. In some embodiments, the phosphoramidite is attached to a moiety attached to a sugar moiety of the nucleoside, e.g., to an optionally substituted triazole ring -P(O)(OR PG Includes )2. In some embodiments, the sugar is a natural sugar. In some embodiments, the sugar is a modified sugar. In some embodiments, the nucleobase is an optionally substituted natural nucleobase. In some embodiments, the nucleobase is an optionally protected A, T, C, G, U, or 5mC (e.g., for oligonucleotide synthesis). In some embodiments, the nucleoside is baseless.

[0278] In some embodiments, the provided technique uses the formula (R PG )2P(O)-L 5E Compounds of -SU(-BA)-OH are provided, wherein each other variable is independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite.

[0279] In some embodiments, the provided technology provides a compound of the formula N or a salt thereof:

[0280] ,

[0281] N

[0282] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite.

[0283] In some embodiments, the provided technology provides a compound of the formula N' or a salt thereof:

[0284] ,

[0285] N'

[0286] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite.

[0287] In some embodiments, the provided technology provides a compound of Formula II or a salt thereof:

[0288] ,

[0289] II

[0290] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite.

[0291] In some embodiments, the provided technology provides a compound of formula II' or a salt thereof:

[0292] ,

[0293] II'

[0294] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite.

[0295] In some embodiments, the provided nucleoside may react with a phosphorus-containing preparation, for example, a compound of formula V or a salt thereof, to provide phosphoramidite.

[0296] In some embodiments, the provided technology provides a compound of Formula III or a salt thereof:

[0297] ,

[0298] III

[0299] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite. In some embodiments, these compounds are utilized, for example, to produce compounds of Formula II or Formula II' or salts thereof.

[0300] In some embodiments, the provided technology provides a compound of formula III' or a salt thereof:

[0301] ,

[0302] III'

[0303] Each variable herein is as independently described herein. In some embodiments, these compounds are utilized, for example, as nucleosides for the production of phosphoramidite. In some embodiments, these compounds are utilized, for example, to produce compounds of Formula II or Formula II' or salts thereof.

[0304] nucleus base

[0305] Various nucleobases may be utilized in phosphoramidites, nucleosides, oligonucleotides, etc., in accordance with the present disclosure. In some embodiments, the nucleobases are natural nucleobases, most commonly occurring being A, T, C, G, and U. In some embodiments, the nucleobases are modified nucleobases in that they are not A, T, C, G, or U. In some embodiments, the nucleobases are optionally substituted A, T, C, G, or U, or substituted tautomers of A, T, C, G, or U. In some embodiments, the nucleobases are optionally substituted A, T, C, G, or U, e.g., 5mC, 5-hydroxymethyl C, etc. In some embodiments, the nucleobases are alkyl-substituted A, T, C, G, or U. In some embodiments, the nucleobases are A. In some embodiments, the nucleobases are T. In some embodiments, the nucleobases are C. In some embodiments, the nucleobases are G. In some embodiments, the nucleobase is U. In some embodiments, the nucleobase is 5mC. In some embodiments, the nucleobase is a substituted A, T, C, G, or U. In some embodiments, the nucleobase is a substituted tautomer of A, T, C, G, or U. In some embodiments, the substitution protects a specific functional group within the nucleobase to minimize undesirable reactions during oligonucleotide synthesis. Suitable techniques for protecting nucleosides in oligonucleotide synthesis are widely known in the art and can be utilized in accordance with the present disclosure.

[0306] In some embodiments, the nucleobase is a natural nucleobase or a modified nucleobase derived from a natural nucleobase. Examples include uracil, thymine, adenine, cytosine, and guanine, each amino group selectively protected by an acyl protecting group, 2-fluorouracil, 2-fluorocytosine, 5-bromouracil, 5-iodouracil, 2,6-diaminopurine, azacytosine, pyrimidine analogs, such as pseudoisocytosine and pseudouracil, and other modified nucleosides, such as 8-substituted purine, xanthine, or hypoxanthine (the latter two are natural degradation products). Specific examples of modified nucleobases are disclosed in Chiu and Rana, RNA, 2003, 9, 1034–1048; Limbach et al., Nucleic Acids Research, 1994, 22, 2183–2196; and Revankar and Rao, Comprehensive Natural Products Chemistry, Vol. 7, 313. In some embodiments, the modified nucleobases are substituted uracil, thymine, adenine, cytosine, or guanine. In some embodiments, the modified nucleobases are functional substitutes of uracil, thymine, adenine, cytosine, or guanine, for example, in terms of hydrogen bonding and / or base pairing. In some embodiments, the nucleobases are optionally substituted uracil, thymine, adenine, cytosine, 5-methylcytosine, or guanine. In some embodiments, the nucleobase is uracil, thymine, adenine, cytosine, 5-methylcytosine, or guanine.

[0307] In some embodiments, the modified base is optionally substituted adenine, cytosine, guanine, thymine, or uracil, or a tautomer thereof. In some embodiments, the modified nucleobase is a modified adenine, cytosine, guanine, thymine, or uracil modified by one or more of the following modifications:

[0308] The nucleobase is modified with one or more optionally substituted groups independently selected from acyl, halogen, amino, azide, alkyl, alkenyl, alkynyl, aryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocyclyl, heteroaryl, carboxyl, hydroxyl, biotin, avidin, streptavidin, substituted silyl, and combinations thereof;

[0309] One or more atoms of the nucleobase are independently replaced by different atoms selected from carbon, nitrogen, and sulfur;

[0310] One or more double bonds of the nucleobase are independently hydrogenated; or

[0311] One or more aryl or heteroaryl rings are independently inserted into the nucleus.

[0312] In some embodiments, the modified nucleus is, for example, the modified nucleus reported in WO2017 / 210647. In some embodiments, the modified nucleus is an extended-size nucleus to which one or more aryl and / or heteroaryl rings, such as a phenyl ring, have been added. Specific examples of modified nucleuses including nucleus substitution are in the Glen Research catalog (Glen Research, Sterling, Virginia); Krueger AT et al., Acc. Chem. Res., 2007, 40, 141-150; Kool, ET, Acc. Chem. Res., 2002, 35, 936-943; Benner SA, et al., Nat. Rev. Genet., 2005, 6, 553-543; Romesberg, FE, et al., Curr. Opin. Chem. Biol., 2003, 7, 723-733; Or described in Hirao, I., Curr. Opin. Chem. Biol., 2006, 10, 622-627. In some embodiments, the expanded-size nucleobase is, for example, the expanded-size nucleobase described in WO2017 / 210647. In some embodiments, the modified nucleobase is a moiety such as a chorine- or porphyrin-derived ring. Specific porphyrin-derived base substitutions have been described, for example, in Morales-Rojas, H and Kool, ET, Org. Lett., 2002, 4, 4377-4380. In some embodiments, the porphyrin-derived ring is, for example, the porphyrin-derived ring described in WO2017 / 219647. In some embodiments, the modified nucleobase is, for example, the modified nucleobase described in WO2017 / 219647. In some embodiments, the modified nucleobase is fluorescent. Examples of such fluorescent modified nucleobases include phenanthrene, pyrene, stilbene, isoxanthin, isoxanthopterin, terphenyl, terthiophene, benzotherthiophene, coumarin, lumazine, tethered stilbene, benzo-uracil, naphtho-uracil, etc., and, for example, those described in WO2017 / 210647.In some embodiments, the nucleus base or modified nucleus base is selected from C5-propin T, C5-propin C, C5-thiazole, phenoxazine, 2-thio-thymine, 5-triazolylphenyl-thymine, diaminopurine, and N2-aminopropylguanine.

[0313] In some embodiments, the modified nucleobase is selected from 5-substituted pyrimidine, 6-azapyrimidine, alkyl or alkynyl-substituted pyrimidine, alkyl-substituted purine, and N-2, N-6 and O-6-substituted purine. In certain embodiments, the modified nucleobase is 2-aminopropyladenine, 5-hydroxymethylcytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-N-methylguanine, 6-N-methyladenine, 2-propyladenine, 2-thiouracil, 2-thiothimine and 2-thiocytosine, 5-propynyl(-C≡C-CH3)uracil, 5-propynylcytosine, 6-azouracil, 6-azocytosine, 6-azothimine, 5-ribosyluracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl, 8-aza and other 8-substituted purines, 5-halo, particularly 5-bromo, It is selected from 5-trifluoromethyl, 5-halouracil, and 5-halocytosine, 7-methylguanine, 7-methyladenine, 2-F-adenine, 2-aminoadenine, 7-deazaguanine, 7-deazadenine, 3-deazaguanine, 3-deazadenine, 6-N-benzoyladenine, 2-N-isobutyrylguanine, 4-N-benzoylcytosine, 4-N-benzoyluracil, 5-methyl 4-N-benzoylcytosine, 5-methyl 4-N-benzoyluracil, universal bases, hydrophobic bases, indiscriminate bases, size-extended bases, and fluorinated bases. In some embodiments, the modified nucleobase is a tricyclic pyrimidine, e.g., 1,3-diazafenoxazine-2-one, 1,3-diazafenothiazine-2-one, or 9-(2-aminoethoxy)-1,3-diazafenoxazine-2-one (G-clamp). In some embodiments, the modified nucleobase is one in which a purine or pyrimidine base is replaced by another heterocyclic ring, e.g., 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine, or 2-pyridone. In some embodiments, the modified nucleobase is US 3687808, The Concise Encyclopedia Of Polymer Science And Engineering, Kroschwitz, JI, Ed.Includes those disclosed in John Wiley & Sons, 1990, 858-859; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613; Sanghvi, YS, Chapter 15, Antisense Research and Applications, Crooke, ST and Lebleu, B., Eds., CRC Press, 1993, 273-288; or Chapters 6 and 15, Antisense Drug Technology, Crooke ST, Ed., CRC Press, 2008, 163-166 and 442-443.

[0314] In some embodiments, the modified nucleobase and the method thereof are US 20030158403, US 3687808, US 4845205, US 5130302, US 5134066, US 5 175273, US 5367066, US 5432272, US 5434257, US 5457187, US 5459255, US 5484908, US 5502177, US 5525711, US 5552540, US 5587469, US 5594 121, US 5596091, US 5614617, US 5645985, US 5681941, US 5750692, US It is listed in 5763588, US 5830653, or US 6005096.

[0315] In some embodiments, the modified nucleobase is substituted. In some embodiments, the modified nucleobase is substituted to contain, for example, a heteroatom, an alkyl group, or a linking moiety (a fluorescent moiety, a biotin or avidin moiety, or a linking moiety to another protein or peptide). In some embodiments, the modified nucleobase is a "universal base" that is not a nucleobase in the most classical sense but functions similarly to a nucleobase. One example of a universal base is 3-nitropyrrole.

[0316] In some embodiments, the nucleoside that may be utilized in the provided technology is a modified nucleobase and / or a modified sugar, e.g., 4-acetylcytidine; 5-(carboxyhydroxymethyl)uridine; 2'-O-methylcytidine; 5-carboxymethylaminomethyl-2-thiouridine; 5-carboxymethylaminomethyluridine; dihydrouridine; 2'-O-methylpseudouridine; beta,D-galactosylqueosin; 2'-O-methylguanosine; N 6 -Isopentenyladenosine; 1-Methyladenosine; 1-Methylpseudouridine; 1-Methylguanosine; 1-Methylinosine; 2,2-Dimethylguanosine; 2-Methyladenosine; 2-Methylguanosine; N 7 -Methylguanosine; 3-Methyl-cytidine; 5-Methylcytidine; 5-Hydroxymethylcytidine; 5-Formylcytosine; 5-Carboxylcytosine; N 6 -Methyladenosine; 7-Methylguanosine; 5-Methylaminoethyluridine; 5-Methoxyaminomethyl-2-Thiouridine; Beta,D-Mannosylqueosin; 5-Methoxycarbonylmethyluridine; 5-Methoxyuridine; 2-Methylthio-N 6 - Isopentenyladenosine; N-((9-beta,D-ribofuranosyl-2-methylthiopurine-6-yl)carbamoyl)threonine; N-((9-beta,D-ribofuranosylpurine-6-yl)-N-methylcarbamoyl)threonine; uridine-5-oxyacetate methyl ester; uridine-5-oxyacetate(v); pseudouridine; queosin; 2-thiocitidine; 5-methyl-2-thiouridine; 2-thiouridine; 4-thiouridine; 5-methyluridine; 2'-O-methyl-5-methyluridine; and 2'-O-methyluridine are included.

[0317] In some embodiments, the nucleobase, for example, the modified nucleobase, comprises one or more biomolecular binding moietyes, such as, for example, an antibody, an antibody fragment, biotin, avidin, streptavidin, a receptor ligand, or a chelating moiety. In other embodiments, the nucleobase is 5-bromouracil, 5-iodouracil, or 2,6-diaminopurine. In some embodiments, the nucleobase comprises a substitution by a fluorescent or biomolecular binding moiety. In some embodiments, the substituent is a fluorescent moiety. In some embodiments, the substituent is biotin or avidin.

[0318] Specific examples of nucleobases and related methods are US 3687808, 4845205, US 513030, US 5134066, US 5175273, US 5367066, US 5432272, US 5457187, US 5457191, US 5459255, US 5484908, US 5502177, US 5525711, US 5552540, US 5587469, US 5594121, US 5596091, US 5614617, US 5681941, US 5750692, US 6015886, US 6147200, US 6166197, US It is listed in 6222025, US 6235887, US 6380368, US 6528640, US 6639062, US 6617438, US 7045610, US 7427672, or US 7495088.

[0319] In some embodiments, the oligonucleotide comprises a nucleobase, a sugar, a nucleoside, and / or internucleotide linkages as described in any of the following: Gryaznov, S; Chen, J.-KJ Am. Chem. Soc. 1994, 116, 3143; Hendrix et al. 1997 Chem. Eur. J. 3: 110; Hyrup et al. 1996 Bioorg. Med. Chem. 4: 5; Jepsen et al. 2004 Oligo. 14: 130-146; Jones et al. J. Org. Chem. 1993, 58, 2983; Koizumi et al. 2003 Nuc. Acids Res. 12: 3267-3273; Koshkin et al. 1998 Tetrahedron 54: 3607-3630; Kumar et al. 1998 Bioo. Med. Chem. Let. 8: 2219-2222; Lauritsen et al. 2002 Chem. Comm. 5: 530-531; Lauritsen et al. 2003 Bioo. Med. Chem. Lett. 13: 253-256; Mesmaeker et al. Angew. Chem., Int. Ed. Engl. 1994, 33, 226; Morita et al. 2001 Nucl. Acids Res. Supp. 1: 241-242; Morita et al. 2002 Bioo. Med. Chem. Lett. 12: 73-76; Morita et al. 2003 Bioo. Med. Chem. Lett. 2211-2226; Nielsen et al. 1997 Chem. Soc. Rev. 73; Nielsen et al. 1997 J. Chem. Soc. Perkins Transl. 1: 3423-3433; Obika et al. 1997 Tetrahedron Lett. 38 (50): 8735-8; Obika et al. 1998 Tetrahedron Lett. 39: 5401-5404; Pallan et al. 2012 Chem. Comm. 48: 8195-8197; Petersen et al. 2003 TRENDS Biotech.21:74-81; Rajwanshi et al. 1999 Chem. Commun. 1395-1396; Schultz et al. 1996 Nucleic Acids Res. 24:2966; Seth et al. 2009 J. Med. Chem. 52:10-13; Seth et al. 2010 J. Med. Chem. 53: 8309-8318; Seth et al. 2010 J. Org. Chem. 75: 1569-1581; Seth et al. 2012 Bio. Med. Chem. Lett. 22: 296-299; Seth et al. 2012 Mol. Ther-Nuc. Acids. 1, e47; Seth, Punit P; Siwkowski, Andrew; Allerson, Charles R; Vasquez, Guillermo; Lee, Sam; Prakash, Thazha P; Kinberger, Garth; Migawa, Michael T; Gaus, Hans; Bhat, Balkrishen; et al. From Nucleic Acids Symposium Series (2008), 52(1), 553-554; Singh et al. 1998 Chem. Comm. 1247-1248; Singh et al. 1998 J. Org. Chem. 63: 10035-39; Singh et al. 1998 J. Org. Chem. 63:6078-6079; Sorensen 2003 Chem. Comm. 2130-2131; Ts'o et al. NY Acad. Sci. 1988, 507, 220; Van Aerschot et al. 1995 Angew. Chem. Int. Ed. Engl. 34: 1338; Vasseur et al. J. Am. Chem. Soc. 1992, 114, 4006; WO 2007090071; or WO 2016 / 079181.

[0320] In some embodiments, the oligonucleotide comprises a modified nucleobase, nucleoside, or nucleotide described in any one of the following: Feldman et al. 2017 J. Am. Chem. Soc. 139: 11427-11433, Feldman et al. 2017 Proc. Natl. Acad. Sci. USA 114: E6478-E6479, Hwang et al. 2009 Nucl. Acids Res. 37: 4757-4763, Hwang et al. 2008 J. Am. Chem. Soc. 130: 14872-14882, Lavergne et al. 2012 Chem. Eur. J. 18: 1231-1239, Lavergne et al. 2013 J. Am. Chem. Soc. 135: 5408-5419, Ledbetter et al. 2018 J. Am. Chem. Soc. 140: 758-765, Malyshev et al. 2009 J. Am. Chem. Soc. 131: 14620-14621, Seo et al. 2009 Chem. Bio. Chem. 10: 2394-2400 (e.g., d3FB, d2Py analogs, d2Py, d3MPy, d4MPy, d5MPy, d34DMPy, d35DMPy, d45DMPy, d5FM, d5PrM, d5SICS, dFEMO, dMMO2, dNaM, dNM01, dTPT3, nucleotides comprising 2'-azido, 2'-chloro, 2'-amino or arabinose sugars, isocarbostyryl-, naphthyl-, and azaindol-nucleotides, and their modifications and derivatives and functionalized versions, e.g., sugars comprising 2'-modifications and / or other modifications, and dMMO2 derivatives having meta-chlorine, -bromine, -iodine, -methyl or -propynyl substituents).

[0321] In some embodiments, the nucleobase comprises at least one optionally substituted ring comprising a heteroatom ring atom. In some embodiments, the nucleobase comprises at least one optionally substituted ring comprising a nitrogen ring atom. In some embodiments, such a ring is aromatic. In some embodiments, the nucleobase is bonded to the sugar through the heteroatom. In some embodiments, the nucleobase is bonded to the sugar through the nitrogen atom. In some embodiments, the nucleobase is bonded to the sugar through the ring nitrogen atom.

[0322] In some embodiments, the oligonucleotide comprises a nucleobase or a modified nucleobase as described in US 10167309, US 11643657, US 11718638, US 11608355 and US 20230089442.

[0323] In some embodiments, the nucleus is an optionally substituted purine base residue. In some embodiments, the nucleus is a protected purine base residue. In some embodiments, the nucleus is an optionally substituted adenine residue. In some embodiments, the nucleus is a protected adenine residue. In some embodiments, the nucleus is an optionally substituted guanine residue. In some embodiments, the nucleus is a protected guanine residue. In some embodiments, the nucleus is an optionally substituted cytosine residue. In some embodiments, the nucleus is a protected cytosine residue. In some embodiments, the nucleus is an optionally substituted thymine residue. In some embodiments, the nucleus is a protected thymine residue. In some embodiments, the nucleus is an optionally substituted uracil residue. In some embodiments, the nucleus is a protected uracil residue. In some embodiments, the nucleus is an optionally substituted 5-methylcytosine residue. In some embodiments, the nucleobase is a protected 5-methylcytosine residue.

[0324] In some embodiments, the provided oligonucleotides are, for example, US 5552540, US 6222025, US 6528640, US 4845205, US 5681941, US 5750692, US 6015886, US 5614617, US 6147200, US 5457187, US 6639062, US 7427672, US 5459255, US 5484908, US 7045610, US 3687808, US 5502177, US 5525711 6235887, US 5175273, US 6617438, US 5594121, US 6380368, US It includes modified nucleobases described in 5367066, US 5587469, US 6166197, US 5432272, US 7495088, US 5134066, or US 5596091. In some embodiments, the nucleobases are described in WO 2020 / 154344, WO 2020 / 154343, WO 2020 / 154342, WO 2020 / 165077, WO 2020 / 201406, WO 2020 / 216637, or WO 2020 / 252376 and may be utilized in accordance with the present disclosure.

[0325] In some embodiments, the nucleobase is a protected base residue, such as that used in the manufacture of oligonucleotides. In some embodiments, the nucleobase is US 2011 / 0294124, US 2015 / 0211006, US 2015 / 0197540, WO 2015 / 107425, WO 2017 / 192679, WO 2018 / 022473, WO 2018 / 098264, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO It is a base residue exemplified in 2019 / 032612 or WO 2020 / 191252.

[0326] sugar

[0327] Various sugars containing modified sugars may be utilized in accordance with the present disclosure, for example, in phosphoramidites, nucleosides, oligonucleotides, etc.

[0328] The most common naturally occurring nucleosides contain a ribose sugar (e.g., in RNA) or a deoxyribose sugar (e.g., in DNA) linked to the nucleobases adenosine (A), cytosine (C), guanine (G), thymine (T), or uracil (U). In some embodiments, for example, in phosphoramidite, nucleoside, oligonucleotide, etc., the sugar is It is a natural DNA sugar (in DNA nucleic acid or oligonucleotide) having the structure, wherein the nucleobase is attached to the 1' position and the 3' and 5' positions are connected to the internucleotide linkage (as will be known to those skilled in the art, if it is at the 5'-terminus of the oligonucleotide, the 5' position may be connected to the 5'-terminal group, and if it is at the 3'-terminus of the oligonucleotide, the 3' position may be connected to the 3'-terminal group (e.g., -OH). In some embodiments, the sugar is a natural RNA sugar (in RNA nucleic acid or oligonucleotide, It has a structure, wherein a nucleobase is attached to the 1' position, and the 3' and 5' positions are connected to the nucleotide linkage (as will be known to those skilled in the art, at the 5'-terminus of the oligonucleotide, the 5' position may be connected to a 5'-terminal group (e.g., -OH), and at the 3'-terminus of the oligonucleotide, the 3' position may be connected to a 3'-terminal group (e.g., -OH). In some embodiments, the sugar is a modified sugar in that it is not a natural DNA sugar or a natural RNA sugar. In particular, the modified sugar may provide improved stability. In some embodiments, the modified sugar may be used to alter and / or optimize one or more hybridization properties. In some embodiments, the modified sugar may be used to alter and / or optimize target nucleic acid recognition. In some embodiments, the modified sugar may be used to optimize Tm. In some embodiments, the modified sugar may be used to improve oligonucleotide activity.

[0329] Sugars can be attached to nucleotide linkages at various positions. For example, nucleotide linkages can be attached to the 2', 3', 4', or 5' positions of the sugar. In some embodiments, as is most common in natural nucleic acids, unless otherwise specified, nucleotide linkages are attached to one sugar at the 5' position and to another sugar at the 3' position.

[0330] In some embodiments, the sugar is an optionally substituted natural DNA or RNA sugar. In some embodiments, the sugar is an optionally substituted is. In some embodiments, the 2' position is optionally substituted. In some embodiments, the sugar is is. In some embodiments, sugar is or It has the structure of, where R 1s , R 2s , R 3s , R 4s , and R 5sEach is independently -H, a suitable substituent, or a suitable sugar modification (e.g., US 9394333, US 9744183, US 9605019, US 9982257, US 20170037399, US 20180216108, US 20180216107, US 9598458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 022473, WO 2018 / 223056, WO 2018 / 223073, WO Those listed in 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 032612, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO 2019 / 032612, WO 2020 / 191252, WO 2021 / 071858 or WO 2022 / 099159, each of these substituents, variations thereof, R 1s , R 2s , R 3s , R 4s , and R 5s Description of, and modified sugars are independently incorporated herein by reference). In some embodiments, R 1s , R 2s , R 3s , R 4s and R 5s Each independently R s and here, each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R', -L s -OR', -L s -SR', -L s -N(R')2, -OL s -OR', -OL s -SR' or -OL s-N(R')2, where each R' is independently as described herein, and each L s is independently covalently bonded or optionally substituted divalent C 1-6 Aliphatic or heteroaliphatic having 1 to 4 heteroatoms; or two R s Take together and bridge -L s - forms. In some embodiments, R' is optionally substituted C 1-10 It is aliphatic. In some embodiments, sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, R 5s is optionally substituted C 1-6 It is aliphatic. In some embodiments, R 5s is optionally substituted C 1-6 It is an alkyl. In some embodiments, R 5s is an optionally substituted methyl. In some embodiments, R 5s is methyl. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the 2'-modified sugar is It has the structure of, and R 2s is a 2'-modification. In some embodiments, the sugar is It has the structure of, and R 2s is -H, halogen, or -OR (R is an optionally substituted C 1-6 (It is aliphatic). In some embodiments, R 2s is -H. In some implementations, R 2s is -F. In some implementations, R 2s is -OMe. In some embodiments, the modified nucleoside is mA, mT, mC, m5mC, mG, mU, etc., where R 2s is -OMe. In some implementations, R 2s is -OCH2CH2OMe. In some implementations, R 2s is -OCH2CH2OH. In some embodiments, the 2'-F modified sugar is It has the structure of. In some embodiments, the 2'-OMe variant per It has the structure of. In some embodiments, per 2'-MOE variant It has the structure of

[0331] In some embodiments, sugar It has the structure of, where R 2s and R 4s is drunk together -L s - forms, and here L s is a covalent bond or an optionally substituted divalent C 1-6 It is aliphatic or heteroaliphatic having 1 to 4 heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, or sulfur). In some embodiments, L s is an optionally substituted C2-O-CH2-C4. In some embodiments, L s is C2-O-CH2-C4. In some embodiments, L s is C2-O-( R It is )-CH(CH2CH3)-C4. In some embodiments, L s is C2-O-( S )-CH(CH2CH3)-C4.

[0332] In some embodiments, sugar It has the structure of, wherein each variable is independently as described herein. In some embodiments, the sugar is It has the structure of, wherein each variable is independently as described herein. In some embodiments, R 5s is -H. In some embodiments, the sugar is It has the structure of, wherein each variable is independently as described herein. In some embodiments, R 3s is -OH. In some embodiments, R 3s is -H. In some embodiments, the sugar is is. In some embodiments, sugar is am.

[0333] In some embodiments, the sugar is optionally substituted and, here, X s is -S-, -Se-, or optionally substituted -CH2-. In some embodiments, the 2' position is optionally substituted. In some embodiments, the sugar is is. In some embodiments, sugar is or It has the structure of, and R 1s , R 2s , R 3s , R 4s , and R 5sEach is independently -H, a suitable substituent, or a suitable sugar modification (e.g., US 9394333, US 9744183, US 9605019, US 9982257, US 20170037399, US 20180216108, US 20180216107, US 9598458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 022473, WO 2018 / 223056, WO 2018 / 223073, WO Those listed in 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 032612, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO 2019 / 032612, WO 2020 / 191252, WO 2021 / 071858 and / or WO 2022 / 099159, each of these substituents, R 1s , R 2s , R 3s , R 4s , and R 5s Description of, and modified sugars are independently incorporated herein by reference). In some embodiments, R 1s , R 2s , R 3s , R 4s and R 5s Each independently R s and here, each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R', -L s -OR', -L s -SR', -L s -N(R')2, -OL s -OR', -OL s -SR' or -OL s-N(R')2, where each R' is independently as described herein, and each L s is independently covalently bonded or optionally substituted divalent C 1-6 Aliphatic or heteroaliphatic having 1 to 4 heteroatoms; or two R s Take together and bridge -L s - forms. In some embodiments, R' is optionally substituted C 1-10 It is aliphatic. In some embodiments, sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, R 5s is optionally substituted C 1-6 It is aliphatic. In some embodiments, R 5s is optionally substituted C 1-6 It is an alkyl. In some embodiments, R 5s is an optionally substituted methyl. In some embodiments, R 5s is methyl. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the sugar is It has the structure of. In some embodiments, the 2'-modified sugar is It has the structure of, and R 2s is a 2'-modification. In some embodiments, the sugar is It has the structure of, and R 2s is -H, halogen, or -OR (R is an optionally substituted C 1-6 (It is aliphatic). In some embodiments, R 2s is -H. In some implementations, R 2s is -F. In some implementations, R 2s is -OMe. In some implementations, R 2s is -OCH2CH2OMe. In some implementations, R 2s is -OCH2CH2OH. In some embodiments, the modified sugar is It has the structure of. In some embodiments, the modified sugar is It has the structure of. In some embodiments, the modified sugar is It has the structure of. In some embodiments, the modified sugar is It has the structure of. In some embodiments, X s is -S-. In some implementations, X s is an optionally substituted -CH2-. In some implementations, X s is -CH2-. In some embodiments, the modified sugar is It has the structure of. In some embodiments, the modified sugar is It has the structure of

[0334] In some embodiments, sugar , , or It has the structure of, where each R 2s -H, -F, -OH, or -OR independently ak and R ak is optionally substituted C 1-6 It is aliphatic, and each other variable is independently as described herein. In some embodiments, R 1s , R 3s , R 4s and R 5s Each is independently -H. In some implementations, R 1s , R 3s and R 4s Each, and R5s One of them is independently -H, and the rest are R 5s C independently 1-6 It is aliphatic. In some embodiments, R 5s The existence of is C 1-6 Aliphatic, e.g., methyl. In some embodiments, R 2s is -H. In some implementations, R 2s is -F. In some implementations, R 2s is --OR ak is. In some implementations, R 2s is -OMe. In some implementations, R 2s is -OCH2CH2CH3. In some implementations, at least one R 2s The existence of is -H. In some embodiments, at least one R 2s The existence of is not -H. In some implementations, X s is -O-. In some implementations, X s is -S-. In some implementations, X s is an optionally substituted -CH2-. In some implementations, X s is -CH2-.

[0335] In some embodiments, sugar It has the structure of, where R 2s and R 4s is drunk together -L s - forms, and here L s is a covalent bond or an optionally substituted divalent C 1-6 It is aliphatic or heteroaliphatic having 1 to 4 heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, or sulfur). In some embodiments, L s is an optionally substituted C2-O-CH2-C4. In some embodiments, L s is C2-O-CH2-C4. In some embodiments, L s is C2-O-( R It is )-CH(CH2CH3)-C4. In some embodiments, Ls is C2-O-( S It is )-CH(CH2CH3)-C4. In some implementations, X s is -S-. In some implementations, X s is an optionally substituted -CH2-. In some implementations, X s is -CH2-. In some implementations, X s is -Se-.

[0336] In some embodiments, sugar It has the structure of, wherein each variable is independently as described herein. In some embodiments, the sugar is It has the structure of, wherein each variable is independently as described herein. In some embodiments, R 5s is -H. In some embodiments, the sugar is It has the structure of, wherein each variable is independently as described herein. In some embodiments, R 3s is -OH. In some embodiments, R 3s is -H. In some implementations, X s is -S-. In some implementations, X s is an optionally substituted -CH2-. In some implementations, X s is -CH2-.

[0337] In some embodiments, a nucleoside containing a modified sugar is Or it has a structure in the form of its salt, BA s is -H or an optionally substituted or protected nucleobase (e.g., BA), and R 2s is as described herein. In some embodiments, R 2s is an -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, BA s is -H. In some implementations, BA s is an optionally substituted or protected nucleobase. In some embodiments, BA sis BA. In some implementations, R 2s is -F. In some embodiments, the nucleoside containing the modified sugar is or has a structure in the form of a salt thereof, and each variable is independently as described herein. In some embodiments, R 2s is -H, -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, R 2s is -H. In some implementations, R 2s is -F. In some embodiments, the nucleoside containing the modified sugar is It has the structure, and each variable is as described herein. In some embodiments, the nucleoside containing the modified sugar is or has a structure in the form of a salt thereof, and each variable is independently as described herein. In some embodiments, R 2s is -H, -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, R 2s is -H. In some implementations, R 2s is -F. In some embodiments, the nucleoside containing the modified sugar is Or having a structure in the form of its salt, where R 2s' is R s and R s , R 2s and BA s Each is as independently described herein. In some embodiments, R 2s and R 2s' Each is independently -H, -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, R 2s is -H. In some implementations, R 2s is -OH. In some embodiments, R 2s is a halogen. In some embodiments, R 2s is -F. In some implementations, R 2sis an optionally substituted C1-C6 alkoxy. In some embodiments, R 2s' is -H. In some implementations, R 2s' is -OH. In some embodiments, R 2s' is a halogen. In some embodiments, R 2s' is -F. In some implementations, R 2s' is an optionally substituted C1-C6 alkoxy. In some embodiments, BA s is -H. In some implementations, BA s is an optionally substituted or protected nucleobase. In some embodiments, BA s is BA. In some embodiments, a nucleobase such as BA is selectively substituted or protected for oligonucleotide synthesis. Such specific nucleosides comprising sugars and nucleosides and their uses are described in WO 2020 / 154342. In some embodiments, the oligonucleotide comprises an arabinoside, a 2'-deoxy-2'-fluoro-arabinoside, a 2'-OR arabinoside, an adeoxycitidine, a DNA-abasic, an RNA-abasic, or a 2'-OR abasic, wherein R is not a hydrogen (e.g., a selectively substituted C 1-6Aliphatic). In some embodiments, 2'-OR is 2'-OMe. In some embodiments, 2'-OR is 2'-MOE. In some embodiments, the oligonucleotide comprises 2'-O-methyl-arabinocytidine (amC). In some embodiments, the oligonucleotide comprises this nucleoside. In some embodiments, the monomer comprises this nucleoside. In some embodiments, the phosphoramidite comprises this nucleoside (in some embodiments, one linkage site (e.g., -CH2- linkage site) is optionally bonded to a substituted -OH, e.g., (-ODMTr), and one linkage site (e.g., ring linkage site) is bonded to an O that is also bonded to the P of the phosphoramidite). In some embodiments, a 5' direction adjacent nucleoside (e.g., N1), an opposite nucleoside (N0), and a 3' direction adjacent nucleoside (e.g., N -1 One or more of ) or each is independently such a nucleoside. In some embodiments, 5'-N1N0N -1 -3' is amCCA. In some embodiments, the sugar is It has the structure of, where each variable is as described herein and C1' is bound to a nucleobase. In some embodiments, the sugar is arabinose. In some embodiments, the sugar is It has the structure of, where C1' is bound to a nucleus.

[0338] In some embodiments, the sugar is optionally substituted and, where, the nucleobase is bound at position 1'. In some embodiments, the sugar is And, here, the nucleobase is bound at position 1'.

[0339] In some embodiments, the sugar is optionally substituted and, here, position a is bound to the nucleobas, and X s is -O-, -S-, -Se-, or optionally substituted -CH2-. In some embodiments, the sugar is is. In some embodiments, the sugar is optionally substituted and, here, position a is bound to the nucleobas, and X s is -O-, -S-, -Se-, or optionally substituted -CH2-. In some embodiments, the sugar is is. In some implementations, X s is -O-. In some implementations, X s is -S-. In some implementations, X s is -Se-. In some implementations, X s is an optionally substituted -CH2-. In some implementations, X s is -CH2-. In some implementations, n is 0. In some implementations, n is 1. In some implementations, n is 2. In some implementations, n is 3.

[0340] In some embodiments, the modified sugar comprises an optionally substituted six-membered ring having 0 to 1 oxygen atom. In some embodiments, the modified sugar comprises an optionally substituted six-membered ring having an oxygen atom. For example, in some embodiments, the modified sugar is optionally substituted It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is optionally substituted It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is optionally substituted It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is optionally substituted It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is optionally substituted It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure of, where, position a is bound to a nucleus base.

[0341] In some embodiments, a nucleoside containing a modified sugar is or has a structure in the form of its salt, R 6s and R 7s Each independently R s Igo, BA s is -H or an optionally substituted or protected nucleobase (e.g., BA), and R s is as independently described herein. In some embodiments, R 6s is -H, -OH, or a halogen, and R 7s is -H, -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, BA s is -H. In some implementations, BA s is an optionally substituted or protected nucleobase. In some embodiments, BA s is BA. In some embodiments, a nucleoside containing a modified sugar is or has a structure in the form of its salt, R 8s and R 9s Each independently R s and R s and BA s Each is as independently described herein. In some embodiments, R 8s is -H or a halogen, and R 9s is -H, -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, the nucleoside containing the modified sugar is or has a structure in the form of its salt, R 10s and R 11s Each independently R s and R s and BA s Each is as independently described herein. In some embodiments, R 10s is -H or a halogen, and R 11s is -H, -OH, a halogen, or an optionally substituted C1-C6 alkoxy. In some embodiments, the nucleoside containing the modified sugar is Or it has a structure in the form of its salt, BA s ...is as described herein. In some embodiments, a nucleoside containing a modified sugar is Or it has a structure in the form of its salt, BA s ...as described herein. Those skilled in the art understand that in some embodiments, nitrogen may be directly bonded to the linked phosphorus. In some embodiments, the halogen is -F. In some embodiments, BA s is -H. In some implementations, BA s is an optionally substituted or protected nucleobase. In some embodiments, BA sis BA. In some embodiments, a nucleobase such as BA is selectively substituted or protected for oligonucleotide synthesis. In some embodiments, the oligonucleotide comprises an alpha-homo-DNA, beta-homo-DNA moiety. In some embodiments, the oligonucleotide comprises an alpha- or beta-homo-DNA sugar. In some embodiments, the oligonucleotide comprises an alpha-homo-DNA sugar. In some embodiments, the oligonucleotide comprises a beta-homo-DNA sugar. These specific nucleosides comprising sugars and nucleobases and their uses are described in WO 2020 / 154343. In some embodiments, the oligonucleotide comprises these nucleosides. In some embodiments, the monomer comprises these nucleosides. In some embodiments, the phosphoramidite comprises such nucleosides (in some embodiments, one linkage site (e.g., -CH2- linkage site) is optionally bonded to a substituted -OH, e.g., -ODMTr, and one linkage site (e.g., ring linkage site) is bonded to P of the phosphoramidite (e.g., if the linking ring atom is N) or to O which is also bonded to P of the phosphoramidite (e.g., if the linking ring atom is C). In some embodiments, a 5' direction adjacent nucleoside (e.g., N1), an opposite nucleoside (N0), and a 3' direction adjacent nucleoside (e.g., N -1 One or more of ) or each is independently such a nucleoside.

[0342] In some embodiments, the modified sugar is It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure of, where position a is bound to a nucleobas. In some embodiments, the modified sugar is It has the structure, where position a is bound to a nucleobase, position b is bound to an internucleoside linkage, and R'' is -H or optionally substituted C 1-6 It is aliphatic. In some embodiments, the modified sugar is It has the structure, where position a is bound to a nucleobase, position b is bound to an internucleotide linkage, and R'' is -H or C 1-6 It is aliphatic. In some embodiments, the modified sugar is It has the structure, where position a is bound to a nucleobase, position b is bound to an internucleotide linkage, and R'' is -H or C 1-6 It is aliphatic. In some embodiments, R" is methyl.

[0343] In some embodiments, a nucleoside containing a modified sugar is or has a structure in the form of a salt thereof, and each variable is as described herein. In some embodiments, a nucleoside containing a modified sugar is or has a structure in the form of a salt thereof, and each variable is as described herein. In some embodiments, a nucleoside containing a modified sugar is or has a structure in the form of a salt thereof, and each variable is as described herein. In some embodiments, a nucleoside containing a modified sugar is or has a structure in the form of its salt, R 12s is R s and R s and BA s Each is as independently described herein. In some embodiments, R 12s is -H, -OH, halogen, optionally substituted C 1-6 Alkyl, optionally substituted C 1-6 Heteroalkyl or optionally substituted C 1-6 It is an alkoxy. In some embodiments, the halogen is -F. In some embodiments, the nucleoside containing the modified sugar is or has a structure in the form of a salt thereof, and each variable is as described herein. In some embodiments, a nucleotide containing a modified sugar is or has a structure in the form of its salt, R 13s is R s and R s and BA s Each is as independently described herein. In some embodiments, R 13s is -H or optionally substituted C1-C6 alkyl. In some embodiments, the nucleoside containing the modified sugar is or has a structure in the form of a salt thereof, and each variable is as described herein. In some embodiments, a nucleotide containing a modified sugar is Or it has a structure in the form of a salt thereof, and each variable is as described herein. In some embodiments, the link is an amide link. In some embodiments, BA s is -H. In some implementations, BA s is an optionally substituted or protected nucleobase. In some embodiments, BA s is BA. In some embodiments, a nucleobase such as BA is optionally substituted or protected for oligonucleotide synthesis. These specific nucleosides and nucleotides containing sugars and nucleobases and their uses are described in WO 2020 / 154344. In some embodiments, the oligonucleotide comprises these nucleosides. In some embodiments, the oligonucleotide comprises these nucleosides (in some embodiments, one linkage site (e.g., -CH2- linkage site) is coupled to an optionally substituted -OH, e.g., (-ODMTr), and one linkage site (e.g., ring linkage site) is coupled to an O that is also coupled to the P of the phosphoramidite). In some embodiments, a 5' direction adjacent nucleoside (e.g., N1), an opposite nucleoside (N0), and a 3' direction adjacent nucleoside (e.g., N -1One or more of ) or each is independently such a nucleoside.

[0344] In some embodiments, the sugar is an acyclic sugar, e.g., a UNA sugar. In some embodiments, the sugar is optionally substituted is. In some embodiments, the 2' position is optionally substituted. In some embodiments, the sugar is is. In some embodiments, sugar is It has the structure of. In some embodiments, R 2s It is -OH. In some embodiments, the sugar is and, where, "*" indicates a carbon atom bonded to a nucleobase. In some embodiments, the sugar is and, where, "*" represents a carbon atom bonded to a nucleobase. In some embodiments, the carbon atom bonded to the nitrogen atom of the nucleobase is R It has an arrangement (e.g., sm18). In some embodiments, the oligonucleotide comprises the sugars described herein.

[0345] In some embodiments, the sugar is optionally substituted and, here, position a is bound to the nucleobas, and X s is -O-, -S-, -Se-, or optionally substituted -CH2-. In some embodiments, the sugar is is. In some implementations, X s is -O-. In some implementations, X s is -S-. In some implementations, X s is -Se-. In some implementations, X s is an optionally substituted -CH2-. In some implementations, X s is -CH2-. In some implementations, n is 0. In some implementations, n is 1. In some implementations, n is 2. In some implementations, n is 3.

[0346] In some embodiments, the sugar is not connected through the 5' and 3' positions. Those skilled in the art understand that for such sugars, 5' may refer to the side / direction toward the 5'-terminus of the oligonucleotide, and 3' may refer to the side / direction toward the 3'-terminus of the oligonucleotide.

[0347] In some implementations, R 1s , R 2s , R 3s , R 4s and R 5s Each independently R s and, here R s independently -H, halogen, -CN, -N3, -NO, -NO2, -L s -R', -L s -Si(R')3, -L s -OR', -L s -SR', -L s -N(R')2, -OL s -R', -OL s -Si(R)3, -OL s -OR', -OL s -SR' or -OL s -N(R')2 and; here L s As described herein, L B And, each other variable is as independently described herein. In some embodiments, R 1s and R 2s Each independently R s is. In some implementations, R s is -H. In some implementations, R s is not -H. In some implementations, L s is a covalent bond. In some embodiments, R 2s and R 4s Each is independently -H, -F, -OR, -N(R)2. In some implementations, R 2s is -H, -F, -OR, -N(R)2. In some implementations, R 4s is -H. In some implementations, R2s and R 4s is 2'-OL s - forms, and here L s is optionally substituted C 1-6 It is an alkylene. In some embodiments, L s is an optionally substituted -CH2-. In some embodiments, L s is an optionally substituted -CH2-.

[0348] In some embodiments, R is hydrogen. In some embodiments, R is not hydrogen. In some embodiments, R is C 1-10 C having 1 to 10 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 It is an optionally substituted group selected from a 5-14 member heteroaryl ring having 1-10 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3-15 member heterocyclic ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur.

[0349] In some embodiments, R is optionally substituted C 1-10 It is aliphatic. In some embodiments, R is optionally substituted C 1-6 It is aliphatic. In some embodiments, R is optionally substituted C 1-6It is an alkyl. In some embodiments, R is optionally substituted hexyl, pentyl, butyl, propyl, ethyl, or methyl. In some embodiments, R is optionally substituted hexyl. In some embodiments, R is optionally substituted pentyl. In some embodiments, R is optionally substituted butyl. In some embodiments, R is optionally substituted propyl. In some embodiments, R is optionally substituted ethyl. In some embodiments, R is optionally substituted methyl. In some embodiments, R is hexyl. In some embodiments, R is pentyl. In some embodiments, R is butyl. In some embodiments, R is propyl. In some embodiments, R is ethyl. In some embodiments, R is methyl. In some embodiments, R is isopropyl. In some embodiments, R is n-propyl. In some embodiments, R is tert-butyl. In some embodiments, R is sec-butyl. In some embodiments, R is n-butyl. In some embodiments, R is -(CH2)2OCH3.

[0350] In some embodiments, R is optionally substituted phenyl. In some embodiments, R is phenyl.

[0351] In some implementations, R 2s is a 2'-variant as described in the present disclosure, and R 4s is -H. In some implementations, R 2s is -OR, and R is not hydrogen. In some embodiments, R 2s is -F. In some implementations, R 2s is -OMe. In some implementations, R 2s is -OCH2CH2CH3. In some implementations, R 2s is selected from -H, -F, and -OR, and R is optionally substituted C 1-6 It is an alkyl. In some embodiments, R 2s is selected from -H, -F, and -OMe.

[0352] In some embodiments, the sugar is a cyclic sugar, e.g., R2s and R 4s Together, it is a sugar that forms an optionally substituted ring as described in the present disclosure. In some embodiments, the sugar is selected from LNA sugars, BNA sugars, cEt sugars, etc. In some embodiments, the crosslink is between the 2' and 4'-carbon atoms (R, taken together with the atom interposed therebetween to form an optionally substituted ring as described herein). 2s and R 4s (corresponding to). In some embodiments, the crosslink is 2'-L a -L b -4' and here L a is -O-, -S- or N(R), and L b The optionally substituted C1-42 is an aliphatic chain, e.g., methylene.

[0353] In some embodiments, the sugar is 2'-OMe, 2'-MOE, 2'-F, LNA (locked nucleic acid) sugar, ENA (ethylene cross-linked nucleic acid) sugar, BNA(NMe) (methylamino cross-linked nucleic acid) sugar, 2'-F ANA (2'-F arabinose), alpha-DNA (alpha-D-ribose), 2' / 5' ODN (e.g., 2' / 5' linked oligonucleotide), Inv (inverted sugar, e.g., inverted deoxyribose), AmR (amino-ribose), ThioR (thio-ribose), HNA (hexos nucleic acid), CeNA (cyclohexene nucleic acid), or MOR (morpholino) sugar.

[0354] After reading the present disclosure, those skilled in the art will understand that various types of sugar variations are known and may be used in accordance with the present disclosure. In some embodiments, the sugar variation is a 2'-variant (e.g., R 2s ) is. In some embodiments, the 2'-modification is 2'-F. In some embodiments, the 2'-modification is 2'-OR, and R is not hydrogen. In some embodiments, the 2'-modification is 2'-OR, and R is optionally a substituted C 1-6It is aliphatic. In some embodiments, the 2'-variant is 2'-OR, and R is optionally a substituted C 1-6 It is an alkyl. In some embodiments, the 2'-modification is 2'-OMe. In some embodiments, the 2'-modification is 2'-MOE. In some embodiments, the 2'-modification is -OL, which links the 2'-carbon of the sugar moiety to another carbon of the sugar moiety. b - or -L b -L b - is. In some embodiments, the 2'-modification is a 2'-OL connecting the 2'-carbon of the sugar moiety to the 4'-carbon of the sugar moiety. b -4' or 2'-L b -L b It is -4'. In some embodiments, the 2'-modification is S-cEt. In some embodiments, the modified sugar is an LNA sugar. In some embodiments, -L b - is -C(R)2-. In some embodiments, the 2'-variant is (C2-OC(R)2-C4), and each R is as independently described in this disclosure. In some embodiments, the 2'-variant is the variant per LNA (C2-O-CH2-C4). In some embodiments, the 2'-variant is (C2-O-CHR-C4), and R is as described in this disclosure. In some embodiments, the 2'-variant is (C2-O-( R )-CHR-C4), where R is as described in the present disclosure and is not hydrogen. In some embodiments, the 2'-modification is (C2-O-( S )-CHR-C4), where R is as described in the present disclosure and is not hydrogen. In some embodiments, R is optionally substituted C 1-6 It is aliphatic. In some embodiments, R is optionally substituted C 1-6 It is an alkyl. In some embodiments, R is an unsubstituted C 1-6 It is alkyl. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, the 2'-modification is (C2-O-CHR-C4), where R is optionally a substituted C 1-6It is aliphatic. In some embodiments, the 2'-variant is (C2-O-CHR-C4), where R is an optionally substituted C 1-6 It is alkyl. In some embodiments, the 2'-modification is (C2-O-CHR-C4), where R is methyl. In some embodiments, the 2'-modification is (C2-O-CHR-C4), where R is ethyl. In some embodiments, the 2'-modification is (C2-O-( R )-CHR-C4), where R is an optionally substituted C 1-6 It is aliphatic. In some embodiments, the 2'-modification is (C2-O-( R )-CHR-C4), where R is an optionally substituted C 1-6 It is an alkyl. In some embodiments, the 2'-modification is (C2-O-( R )-CHR-C4), where R is methyl. In some embodiments, the 2'-modification is (C2-O-( R )-CHR-C4), where R is ethyl. In some embodiments, the 2'-modification is (C2-O-( S )-CHR-C4), where R is an optionally substituted C 1-6 It is aliphatic. In some embodiments, the 2'-modification is (C2-O-( S )-CHR-C4), where R is an optionally substituted C 1-6 It is an alkyl. In some embodiments, the 2'-modification is (C2-O-( S )-CHR-C4), where R is methyl. In some embodiments, the 2'-modification is (C2-O-( S )-CHR-C4), where R is ethyl. In some embodiments, the 2'-modification is C2-O-( R )-CH(CH2CH3)-C4. In some embodiments, the 2'-variant is C2-O-( SIt is )-CH(CH2CH3)-C4. In some embodiments, the sugar is a natural DNA sugar. In some embodiments, the sugar is a natural RNA sugar. In some embodiments, the sugar is an optionally substituted natural DNA sugar. In some embodiments, the sugar is an optionally substituted natural DNA sugar at 2'. In some embodiments, the sugar is a 2'-substituted (2'-modified) natural DNA sugar. In some embodiments, the sugar is a 2'-modified (2'-modified) natural DNA sugar.

[0355] In some embodiments, the sugar is optionally substituted ribose or deoxyribose. In some embodiments, the sugar is optionally modified ribose or deoxyribose, and one or more hydroxyl groups of the ribose or deoxyribose moiety are optionally and independently replaced with a halogen, R', -N(R')2, -OR', or -SR', each R' as described herein. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with a halogen, R', -N(R')2, -OR', or -SR', each R' is independently described herein. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with a halogen. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with one or more -F groups. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with -OR', and each R' is independently described in the present disclosure. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with -OR', and each R' is independently optionally substituted C1-C6 aliphatic. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with -OR', and each R' is independently optionally substituted C1-C6 alkyl. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with -OMe. In some embodiments, the sugar is optionally substituted deoxyribose, and the 2' position of the deoxyribose is optionally substituted with -O-methoxyethyl.

[0356] In some embodiments, the provided oligonucleotide comprises one or more modified sugars. In some embodiments, the provided oligonucleotide comprises one or more modified sugars and one or more natural sugars.

[0357] Examples of cyclic sugars include alpha-L-methyleneoxy(4'-CH2-O-2') LNA, beta-D-methyleneoxy(4'-CH2-O-2') LNA, ethyleneoxy(4'-(CH2)2-O-2') LNA, aminooxy(4'-CH2-ON(R)-2') LNA, and oxyamino(4'-CH2-N(R)-O-2') LNA sugars. In some embodiments, the cyclic sugar, e.g., LNA or BNA sugar, is a sugar having at least one crosslink between two sugar carbons. In some embodiments, the cyclic sugar of the nucleoside may have a stereochemical arrangement of alpha-L-ribofuranose or beta-D-ribofuranose.

[0358] In some embodiments, the cyclic sugar may be further defined by an isomer arrangement. For example, a sugar containing a 4'-(CH2)-O-2' crosslink may exist as an alpha-L arrangement or a beta-D arrangement. In some embodiments, the 4'-2' crosslinkers are -L-4'-(CH2)-O-2', bD-4'-CH2-O-2', 4'-(CH2)2-O-2', 4'-CH2-ON(R')-2', 4'-CH2-N(R')-O-2', 4'-CH(R')-O-2', 4'-CH(CH3)-O-2', 4'-CH2-S-2', 4'-CH2-N(R')-2', 4'-CH2-CH(R')-2', 4'-CH2-CH(CH3)-2', and 4'-(CH2)3-2', respectively, where each R' is as described in the present disclosure. In some embodiments, R' is -H, a protecting group, or optionally a substituted C1-C 12 It is an alkyl. In some embodiments, R' is -H or optionally a substituted C1-C 12 It is an alkyl.

[0359] In some embodiments, the cyclic sugar is alpha-L-methyleneoxy (4'-CH2-O-2') BNA, beta-D-methyleneoxy (4'-CH2-O-2') BNA, ethyleneoxy (4'-(CH2)2-O-2') BNA, aminooxy (4'-CH2-ON(R)-2') BNA, oxyamino (4'-CH2-N(R)-O-2') BNA, methyl(methyleneoxy) (4'-CH(CH3)-O-2') BNA (also called bound ethyl or cEt), methylene-thio (4'-CH2-S-2') BNA, methylene-amino (4'-CH2-N(R)-2') BNA, methyl carboncyclic (4'-CH2-CH(CH3)-2') BNA, propylene carboncyclic (4'-(CH2)3-2') BNA, or vinyl It is BNA's party.

[0360] In some embodiments, the sugar variant is a variant described in US 9006198. In some embodiments, the modified sugar is described in US 9006198. In some embodiments, the present invention relates to US 9394333, US 9744183, US 9605019, US 9982257, US 20170037399, US 20180216108, US 20180216107, US 9598458, WO 2017 / 062862, WO 2018 / 067973, WO 2017 / 160741, WO 2017 / 192679, WO 2017 / 210647, WO 2018 / 098264, WO 2018 / 022473, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO Variants described in 2018 / 237194, WO 2019 / 032607, WO 2019 / 032612, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO 2019 / 032612, WO 2020 / 191252, WO 2021 / 071858, and / or WO 2022 / 099159, each sugar variation and modified sugar are independently incorporated herein by reference.

[0361] 일부 구현예에서, 변형된 당은 US 5658873, US 5118800, US 5393878, US 5514785, US 5627053, US 7034133;7084125, US 7399845, US 5319080, US 5591722, US 5597909, US 5466786, US 6268490, US 6525191, US 5519134, US 5576427, US 6794499, US 6998484, US 7053207, US 4981957, US 5359044, US 6770748, US 7427672, US 5446137, US 6670461, US 7569686, US 7741457, US 8022193, US 8030467, US 8278425, US 5610300, US 5646265, US 8278426, US 5567811, US 5700920, US 8278283, US 5639873, US 5670633, US 8314227, US 2008 / 0039618, US 2009 / 0012281, WO 2021 / 030778, WO 2020 / 154344, WO 2020 / 154343, WO 2020 / 154342, WO 2020 / 165077, WO 2020 / 201406, WO 2020 / 216637, 또는 WO 2020 / 252376에 기재된 것이다.

[0362] In some embodiments, the sugar modification is 2'-OMe, 2'-MOE, 2'-LNA, 2'-F, 5'-vinyl, or S-cEt. In some embodiments, the modified sugar is the sugar of FRNA, FANA, or morpholino. In some embodiments, the oligonucleotide comprises a nucleic acid analog, e.g., GNA, LNA, PNA, TNA, F-HNA (F-THP or 3'-fluorotetrahydropyran), MNA (mannitol nucleic acid, e.g., Leumann 2002 Bioorg. Med. Chem. 10: 841-854), ANA (anitol nucleic acid), or morpholino, or a part thereof. In some embodiments, the sugar is the same as the sugar in flexible nucleic acid or serinol nucleic acid. In some embodiments, the sugar modification replaces the natural sugar with another cyclic or acyclic moiety. Examples of such moiety are widely known in the art (e.g., those used in morpholino, glycol nucleic acids, etc.) and may be used in accordance with the present disclosure. As understood by those skilled in the art, when used with a modified sugar, in some embodiments the internucleotide linkage may be modified, for example, as in morpholino, PNA, etc. In some embodiments, the sugar is an (R)-GNA sugar. In some embodiments, the sugar is an (S)-GNA sugar. In some embodiments, the nucleoside having a GNA sugar is N -1 , is utilized as N0 and / or N1. In some embodiments, N0 is a nucleoside having a GNA sugar. In some embodiments, the sugar is a bicyclic sugar. In some embodiments, the sugar is an LNA sugar. In some embodiments, the sugar is an acyclic sugar. In some embodiments, the sugar is an UNA sugar. In some embodiments, the nucleoside having an UNA sugar is N -1 , is utilized as N0 and / or N1. In some embodiments, N0 is a nucleoside having an UNA sugar. In some embodiments, the nucleoside is baseless. In some embodiments, the baseless sugar is N -1, is utilized as N0 and / or N1. In some embodiments, N0 is a nucleoside having a baseless sugar.

[0363] In some embodiments, the sugar is a 6'-modified bicyclic sugar having (R) or (S)-chirality at position 6 (e.g., as described in US 7399845). In some embodiments, the sugar is a 5'-modified bicyclic sugar having (R) or (S)-chirality at position 5 (e.g., as described in US 20070287831).

[0364] In some embodiments, the modified sugar is -F; -CF3, -CN, -N3, -NO, -NO2, -OR', -SR' or -N(R')2 (wherein each R' is independently described in the present disclosure); -O-(C1-C 10 alkyl), -S-(C1-C 10 alkyl), -NH-(C1-C 10 alkyl) or -N(C1-C 10 Alkyl)2; -O-(C2-C 10 alkenyl), -S-(C2-C 10 alkenyl), -NH-(C2-C 10 alkenyl) or -N(C2-C 10 Alkenyl)2; -O-(C2-C 10 alkynyl), -S-(C2-C 10 alkynyl), -NH-(C2-C 10 alkynyl) or -N(C2-C 10 alkynyl)2; or -O--(C1-C 10 Alkylene)-O--(C1-C 10 alkyl), -O-(C1-C 10 Alkylene)-NH-(C1-C 10 alkyl) or -O-(C1-C 10 Alkylene)-NH(C1-C 10 Alkyl)2, -NH-(C1-C 10 Alkylene)-O-(C1-C 10 alkyl) or -N(C1-C 10 Alkyl)-(C1-C 10 Alkylene)-O-(C1-C 10It includes one or more substituents independently selected from alkyl)(wherein each alkyl, alkylene, alkenyl, and alkynyl is independently and optionally substituted) (typically one substituent, and often in the axial position) at the 2' position. In some embodiments, the substituent is -O(CH2) n OCH3, -O(CH2) n The modified sugar is NH2, MOE, DMAOE, or DMAEOE, where n is 1 to about 10. In some embodiments, the modified sugar is that described in WO 2001 / 088198; and Martin et al., Helv. Chim. Acta, 1995, 78, 486-504. In some embodiments, the modified sugar comprises one or more groups selected from substituted silyl groups, RNA cleavers, reporter groups, fluorescent labels, intercalators, groups for improving the pharmacokinetic properties of nucleic acids, groups for improving the pharmacodynamic properties of nucleic acids, or other substituents having similar properties. In some embodiments, the modification is made at one or more of the 2', 3', 4', or 5' positions, including the 3' position of the sugar on the 3'-terminal nucleoside or the 5' position of the 5'-terminal nucleoside.

[0365] In some embodiments, the 2'-OH of ribose is -H, -F; -CF3, -CN, -N3, -NO, -NO2, -OR', -SR' or -N(R')2 (wherein each R' is independently described in this disclosure); -O-(C1-C 10 alkyl), -S-(C1-C 10 alkyl), -NH-(C1-C 10 alkyl) or -N(C1-C 10 Alkyl)2; -O-(C2-C 10 alkenyl), -S-(C2-C 10 alkenyl), -NH-(C2-C 10 alkenyl) or -N(C2-C 10 Alkenyl)2; -O-(C2-C 10 alkynyl), -S-(C2-C 10alkynyl), -NH-(C2-C 10 alkynyl) or -N(C2-C 10 alkynyl)2; or -O-(C1-C 10 Alkylene)-O--(C1-C 10 alkyl), -O-(C1-C 10 Alkylene)-NH-(C1-C 10 alkyl) or -O-(C1-C 10 Alkylene)-NH(C1-C 10 Alkyl)2, -NH-(C1-C 10 Alkylene)-O-(C1-C 10 alkyl) or -N(C1-C 10 Alkyl)-(C1-C 10 Alkylene)-O-(C1-C 10 It is replaced with a group selected from alkyl (wherein each alkyl, alkylene, alkenyl, and alkynyl is independently and optionally substituted). In some embodiments, 2'-OH is replaced with -H (deoxyribose). In some embodiments, 2'-OH is replaced with -F. In some embodiments, 2'-OH is replaced with -OR'. In some embodiments, 2'-OH is replaced with -OMe. In some embodiments, 2'-OH is replaced with -OCH2CH2OMe.

[0366] In some embodiments, the variant is a 2'-variant. Commonly used 2'-variants include, but are not limited to, 2'-OR, where R is not hydrogen and is as described in this disclosure. In some embodiments, the variant is 2'-OR, and R is optionally a substituted C 1-6 It is aliphatic. In some embodiments, the variant is 2'-OR, and R is optionally a substituted C 1-6It is alkyl. In some embodiments, the modification is 2'-OMe. In some embodiments, the modification is 2'-MOE. In some embodiments, the 2'-modification is S-cEt. In some embodiments, the modified sugar is an LNA sugar. In some embodiments, the 2'-modification is -F. In some embodiments, the 2'-modification is FANA. In some embodiments, the 2'-modification is FRNA. In some embodiments, the sugar modification is a 5'-modification, e.g., 5'-Me. In some embodiments, the sugar modification changes the size of the sugar ring. In some embodiments, the sugar modification is the sugar moiety in FHNA.

[0367] In some embodiments, the sugar modification replaces the sugar moiety with another cyclic or acyclic moiety. Examples of such moiety are widely known in the art and include, but are not limited to, those used in morpholino (which optionally has a phosphorodiamidate linkage), glycol nucleic acid, etc.

[0368] The modified sugar comprises a cyclobutyl or cyclopentyl moiety instead of a pentofuranosyl sugar. Representative examples of such modified sugars include those described in US 4,981,957, US 5,118,800, US 5,319,080, or US 5,359,044. In some embodiments, the oxygen atom in the ribose ring is replaced with nitrogen, sulfur, selenium, or carbon. In some embodiments, -O- is replaced with -N(R')-, -S-, -Se-, or -C(R')2-. In some embodiments, the modified sugar is a modified ribose in which the oxygen atom in the ribose ring is replaced with nitrogen, and the nitrogen is optionally substituted with an alkyl group (e.g., methyl, ethyl, isopropyl, etc.).

[0369] An example of a modified sugar is glycerol, which is part of glycerol nucleic acid (GNA), as described, for example, in Zhang, R et al., J. Am. Chem. Soc., 2008, 130, 5846-5847; Zhang L, et al., J. Am. Chem. Soc., 2005, 127, 4174-4175 and Tsai CH et al., PNAS, 2007, 14598-14603.

[0370] Flexible nucleic acids (FNA) are based on mixed acetal aminals of formylglycerol, as described, for example, in Joyce GF et al., PNAS, 1987, 84, 4398-4402 and Heuberger BD and Switzer C, J. Am. Chem. Soc., 2008, 130, 412-413.

[0371] In some embodiments, one or more hydroxyl groups in the sugar are optionally and independently replaced with a halogen, R' -N(R')2, -OR', or -SR', and each R' is independently described in the present disclosure.

[0372] In some embodiments, the modified nucleoside is the modified nucleoside described in WO 2018 / 022473, WO 2018 / 098264, WO 2018 / 223056, WO 2018 / 223073, WO 2018 / 223081, WO 2018 / 237194, WO 2019 / 032607, WO 2019 / 055951, WO 2019 / 075357, WO 2019 / 200185, WO 2019 / 217784, WO 2019 / 032612, WO 2020 / 191252, WO 2021 / 071858, and / or WO 2022 / 099159, each of these modified nucleosides is independently incorporated herein by reference.

[0373] In some embodiments, the sugar modification is 5'-vinyl (R or S), 5'-methyl (R or S), 2'-SH, 2'-F, 2'-OCH3, 2'-OCH2CH3, 2'-OCH2CH2F, or 2'-O(CH2) 20 CH3. In some embodiments, the substituent at the 2' position, e.g., the 2'-modified, is allyl, amino, azido, thio, -O-allyl, -O-C1-C 10 Alkyl, -OCF3, -OCH2F, -O(CH2)2SCH3, -O(CH2)2ON(R')2, -OCH2C(O)N(R')2, and -OCH2C(O)N(R')(CH2)2N(R')2, wherein each allyl, amino, and alkyl group is optionally substituted, and each R' is independently as described in the present disclosure. In some embodiments, each R' is independently -H or an optionally substituted C1-C 10 It is an alkyl.

[0374] In some embodiments, the bicyclic sugar is crosslinked between two sugar carbons, for example, between the 4' and 2' ribosyl ring carbon atoms, for example, -L b -L b Includes -, -L- etc. In some embodiments, the crosslink is 4'-(CH2)-O-2' (e.g., per LNA), 4'-(CH2)-S-2', 4'-(CH2)2-O-2' (e.g., per ENA), 4'-CH(R')-O-2' (e.g., 4'-CH(CH3)-O-2', 4'-CH(CH2OCH3)-O-2' and examples of US 7399845, etc.), 4'-CH(R')2-O-2' (e.g., 4'-C(CH3)(CH3)-O-2' and examples of WO 2009006478, etc.), 4'-CH2-N(OR')-2' (e.g., 4'-CH2-N(OCH3)-2', examples of WO 2008150729, etc.), 4'-CH2-ON(R')-2' (e.g., 4'-CH2-ON(CH3)-2', example in US 20040171570, etc.), 4'-CH2-N(R')-O-2'[e.g., where R is -H, C1-C 12It is an alkyl or protecting group (e.g., see US 7427672), 4'-C(R')2-C(H)(R')-2' (e.g., 4'-CH2-C(H)(CH3)-2', example in Chattopadhyaya et al., J. Org. Chem., 2009, 74, 118-134, etc.) or 4'-C(R')2-C(=C(R')2)-2' (e.g., 4'-CH2-C(=CH2)-2', example in WO 2008154401, etc.).

[0375] In some embodiments, the sugar is tetrahydropyran or THP sugar. In some embodiments, the modified nucleoside is tetrahydropyran nucleoside or THP nucleoside, which is a nucleoside having a hexavalent tetrahydropyran sugar substituted with a pentopranosyl residue of a common natural nucleoside. The THP sugar and / or nucleoside includes those used in hexitol nucleic acid (HNA), anitol nucleic acid (ANA), mannitol nucleic acid (MNA) (e.g., Leumann, Bioorg. Med. Chem., 2002, 10, 841-854) or fluoro HNA (F-HNA).

[0376] In some embodiments, the sugar comprises a ring having more than 5 atoms and / or more than 1 heteroatom, e.g., morpholino sugars, e.g., Braasch et al., Biochemistry, 2002, 41, 4503-4510; US 5698685; US 5166315; US 5185444; US 5034506; etc.

[0377] In some embodiments, the nucleoside has a hexavalent cyclohexenyl group instead of the pentopranosyl residue of the naturally occurring nucleoside. Examples of cyclohexenyl nucleosides and their preparation and use are, for example, WO 2010036696; Robeyns et al., J. Am. Chem. Soc., 2008, 130(6), 1979–1984; Horvath et al., Tetrahedron Letters, 2007, 48, 3621–3623; Nauwelaerts et al., J. Am. Chem. Soc., 2007, 129(30), 9340–9348; Gu et al., Nucleosides, Nucleotides & Nucleic Acids, 2005, 24(5–7), 993–998; Nauwelaerts et al., Nucleic Acids Research, 2005, 33(8), 2452-2463; Robeyns et al., Acta Crystallographica, Section F: Structural Biology and Crystallization Communications, 2005, F61(6), 585-586; Gu et al., Tetrahedron, 2004, 60(9), 2111-2123; Gu et al., Oligonucleotides, 2003, 13(6), 479-489; Wang et al., J. Org. Chem., 2003, 68, 4499-4505; Verbeure et al., Nucleic Acids Research, 2001, 29(24), 4941-4947; Wang et al., J. Org. Chem., 2001, 66, 8478-82; Wang et al., Nucleosides, Nucleotides & Nucleic Acids, 2001, 20(4-7), 785-788; Wang et al., J. Am. Chem., 2000, 122, 8595-8602; WO 2006047842; WO 2001049687; etc.

[0378] Many monocyclic, bicyclic, and tricyclic ring systems are suitable as sugar substitutes (modified sugars) and can be used in accordance with the present disclosure. See, for example, Leumann, Christian J. Bioorg. & Med. Chem., 2002, 10, 841-854. These ring systems may have their properties and / or activity further enhanced through various additional substitutions.

[0379] In some embodiments, the 2'-modified sugar is a furanosyl sugar modified at the 2' position. In some embodiments, the 2'-modification is a halogen, -R' (R' is not -H), -OR' (R' is not -H), -SR', -N(R')2, optionally substituted -CH2-CH=CH2, optionally substituted alkenyl, or optionally substituted alkynyl. In some embodiments, the 2'-modification is -O[(CH2) n O] m CH3, -O(CH2) n NH2, -O(CH2) n CH3, -O(CH2) n F, -O(CH2) n ONH2, -OCH2C(=O)N(H)CH3 and -O(CH2) n ON[(CH2) n Selected from CH3]2, where each n and m is independently 1 to about 10. In some embodiments, the 2'-variant is optionally a substituted C1-C 12alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted alkaryl, optionally substituted aralkyl, optionally substituted -O-alkaryl, optionally substituted -O-aralkyl, -SH, -SCH3, -OCN, -Cl, -Br, -CN, -F, -CF3, -OCF3, -SOCH3, -SO2CH3, -ONO2, -NO2, -N3, -NH2, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkaryl, optionally substituted aminoalkylamino, optionally substituted polyalkylamino, substituted silyl, reporter group, intercalator, group for improving pharmacokinetic properties, group for improving pharmacodynamic properties, and other substituents. In some embodiments, the 2'-modification is a 2'-MOE modification (e.g., Baker et al., J. Biol. Chem., 1997, 272, 11944-12000). In some cases, the 2'-MOE modification has been reported to have improved binding affinity compared to the unmodified sugar and some other modified nucleosides, such as 2'-O-methyl, 2'-O-propyl, and 2'-O-aminopropyl. Oligonucleotides with 2'-MOE modifications have also been reported to be capable of inhibiting gene expression with promising characteristics for in vivo use (see, e.g., Martin, Helv. Chim. Acta, 1995, 78, 486-504; Altmann et al., Chimia, 1996, 50, 168-176; Altmann et al., Biochem. Soc. Trans., 1996, 24, 630-637; and Altmann et al., Nucleosides Nucleotides, 1997, 16, 917-926; et al.).

[0380] In some embodiments, the 2'-modified or 2'-substituted sugar or nucleoside is a sugar or nucleoside comprising a substituent other than -H (which is generally not considered a substituent) or -OH at the 2' position of the sugar. In some embodiments, the 2'-modified sugar is a bicyclic sugar comprising a crosslinker connecting two carbon atoms of the sugar ring (one of which is the 2' carbon). In some embodiments, the 2'-modified is non-crosslinked, e.g., allyl, amino, azido, thio, optionally substituted -O-allyl, optionally substituted -O-C1-C 10 Alkyl, -OCF3, -O(CH2)2OCH3, 2'-O(CH2)2SCH3, -O(CH2)2ON(R m )(R n ) or -OCH2C(=O)N(R m )(R n ) and, where each R m and R n is independently -H or optionally substituted C1-C 10 It is an alkyl.

[0381] Certain modified sugars, their manufacture and use are described in US 4981957, US 5118800, US 5319080, US 5359044, US 5393878, US 5446137, US 5466786, US 5514785, US 5519134, US 5567811, US 5576427, US 5591722, US 5597909, US 5610300, US 5627053, US 5639873, US 5646265, US 5670633, US 5700920, US 5792847, US 6600032, and WO 2005121371.

[0382] In some embodiments, the sugar is a sugar of N-methanocarba, LNA, cMOE BNA, cEt BNA, α-L-LNA or related analogs, HNA, Me-ANA, MOE-ANA, Ara-FHNA, FHNA, R-6'-Me-FHNA, S-6'-Me-FHNA, ENA, or c-ANA. In some embodiments, the modified internucleotide linkage is a C3-amide (e.g., a sugar having an amide modification attached to C3', Mutisya et al. 2014 Nucleic Acids Res. June 1, 2014; 42(10): 6542-6551), formacetal, thioformacetal, MMI [e.g., methylene(methylimino), Peoc'h et al. 2006 Nucleosides and Nucleotides 16 (7-9)], PMO (phosphorodiamidate-linked morpholino) linkage (which links two sugars), or PNA (peptide nucleic acid) linkage. In some embodiments, examples of the internucleotide linkage and / or sugar are Allerson et al. 2005 J. Med. Chem. 48: 901-4; BMCL 2011 21: 1122; BMCL 2011 21: 588; BMCL 2012 22: 296; Chattopadhyaya et al. 2007 J. Am. Chem. Soc. 129:8362; Chem. Bio. Chem. 2013 14:58; Curr. Prot. Nucl. Acids Chem. 2011 1.24.1; Egli et al. 2011 J. Am. Chem. Soc. 133: 16642; Hendrix et al. 1997 Chem. Eur. J. 3: 110; Hyrup et al. 1996 Bioorg. Med. Chem. 4: 5; Imanishi 1997 Tet. Lett. 38:8735; J. Am. Chem. Soc. 1994, 116, 3143; J. Med. Chem. 2009 52: 10; J. Org. Chem. 2010 75: 1589; Jepsen et al. 2004 Oligo. 14: 130-146; Jones et al. J. Org. Chem.1993, 58, 2983; Jung et al. 2014 ACIEE 53: 9893; Kodama et al. 2014 AGDS; Koizumi 2003 BMC 11: 2211; Koizumi et al. 2003 Nuc. Acids Res. 12: 3267-3273; Koshkin et al. 1998 Tetrahedron 54: 3607-3630; Kumar et al. 1998 Bioo. Med. Chem. Let. 8: 2219-2222; Lauritsen et al. 2002 Chem. Comm. 5: 530-531; Lauritsen et al. 2003 Bioo. Med. Chem. Lett. 13: 253-256; Lima et al. 2012 Cell 150: 883-894; Mesmaeker et al. Angew. Chem., Int. Ed. Engl. 1994, 33, 226; Migawa et al. 2013 Org. Lett. 15: 4316; Mol. Ther. Nucl. Acids 2012 1: e47; Morita et al. 2001 Nucl. Acids Res. Supp. 1: 241-242; Morita et al. 2002 Bioo. Med. Chem. Lett. 12: 73-76; Morita et al. 2003 Bioo. Med. Chem. Lett. 2211-2226; Murray et al. 2012 Nucl. Acids Res. 40: 6135; Nielsen et al. 1997 Chem. Soc. Rev. 73; Nielsen et al. 1997 J. Chem. Soc. Perkins Transl. 1: 3423-3433; Obika et al. 1997 Tetrahedron Lett. 38 (50): 8735-8; Obika et al. 1998 Tetrahedron Lett. 39: 5401-5404; Obika et al. 2008 J. Am. Chem. Soc. 130: 4886; Obika et al. 2011 Org. Lett. 13:6050; Oestergaard et al. 2014 JOC 79: 8877; Pallan et al. 2012 Biochem.51: 7; Pallan et al. 2012 Chem. Comm. 48: 8195-8197; Petersen et al. 2003 TRENDS Biotech. 21: 74-81; Prakash et al. 2010 J. Med. Chem. 53: 1636; Prakash et al. 2015 Nucl. Acids Res. 43: 2993-3011; Prakash et al. 2016 Bioorg. Med. Chem. Lett. 26: 2817-2820; Rajwanshi et al. 1999 Chem. Commun. 1395-1396; Schultz et al. 1996 Nucleic Acids Res. 24: 2966; Seth et al. 2008 Nucl. Acid Sym. Ser. 52: 553; Seth et al. 2009 J. Med. Chem. 52: 10-13; Seth et al. 2010 J. Am. Chem. Soc. 132: 14942; Seth et al. 2010 J. Med. Chem. 53: 8309-8318; Seth et al. 2010 J. Org. Chem. 75: 1569-1581; Seth et al. 2011 BMCL 21: 4690; Seth et al. 2012 Bioo. Med. Chem. Lett. 22: 296-299; Seth et al. 2012 Mol. Ther-Nuc. Acids. 1, e47; Seth et al., Nucleic Acids Symposium Series (2008), 52(1), 553-554; Singh et al. 1998 Chem. Comm. 1247-1248; Singh et al. 1998 J. Org. Chem. 63: 10035-39; Singh et al. 1998 J. Org. Chem. 63: 6078-6079; Sorensen 2003 Chem. Comm. 2130-2131; Starrup et al. 2010 Nucl. Acids Res. 38: 7100; Swayze et al. 2007 Nucl. Acids Res. 35: 687; Ts'o et al. Ann. NY Acad. Sci.It is described in 1988, 507, 220; Van Aerschot et al. 1995 Angew. Chem. Int. Ed. Engl. 34: 1338; Vasseur et al. J. Am. Chem. Soc. 1992, 114, 4006; WO 2007090071; WO 2016079181; US ​​6326199; US 6066500; or US 6440739.

[0383] Various additional sugars useful for preparing oligonucleotides or analogs thereof are known in the art and can be utilized in accordance with the present disclosure.

[0384] Links between nucleotides

[0385] Above all, the present disclosure provides various internucleotide linkages, which include various modified internucleotide linkages that can be utilized with other structural elements, e.g., various sugars as described herein, to provide oligonucleotides and compositions thereof.

[0386] Various internucleotide linkages may be used in oligonucleotides according to the present disclosure. In some embodiments, the oligonucleotide comprises one or more types of internucleotide linkages. In some embodiments, the oligonucleotide comprises two or more types of internucleotide linkages. In some embodiments, the oligonucleotide comprises at least three types of internucleotide linkages. In some embodiments, the linkage contains an atom in which the oxygen atom is not bonded to the backbone sugar or is bonded to an oxygen atom that is not part of the backbone sugar (“PO linkage”, e.g., natural phosphate linkage). In some embodiments, the linkage contains an atom in which the sulfur atom is not bonded to the backbone sugar or is bonded to a sulfur atom that is not part of the backbone sugar (“PS linkage”, e.g., phosphorothioate internucleotide linkage). In some embodiments, the linkage contains an atom in which the nitrogen atom is not bonded to the backbone sugar or is bonded to a nitrogen atom that is not part of the backbone sugar (“PN linkage”, e.g., n001). In some embodiments, the oligonucleotide comprises one or more PS linkages. In some embodiments, the oligonucleotide comprises one or more PO links. In some embodiments, the oligonucleotide comprises one or more PN links. In some embodiments, the oligonucleotide comprises one or more PS links and one or more PO links. In some embodiments, the oligonucleotide comprises one or more PS links and one or more PN links. In some embodiments, the oligonucleotide comprises one or more PS links, one or more PN links, and one or more PO links. In some embodiments, the PS link is a phosphorothioate link. In some embodiments, each PS link is independently a phosphorothioate link. In some embodiments, the PO link is a natural phosphate link. In some embodiments, each PO link is independently a natural phosphate link. In some embodiments, the PN link is a phosphoryl guanidine link.In some embodiments, each PN link is independently a phosphoryl guanidine link.

[0387] In some embodiments, the provided oligonucleotide comprises both one or more modified internucleotide linkages and one or more natural phosphate linkages. As is widely known to those skilled in the art, natural phosphate linkages are widely found in natural DNA and RNA molecules; they have the structure -OP(O)(OH)O-, link sugars in nucleosides of DNA and RNA, and may exist in various salt forms; for example, at physiological pH (about 7.4), natural phosphate linkages exist mainly in salt forms, and the anion is -OP(O)(OH) - )O-. Modified internucleotide linkages or non-natural phosphate linkages are internucleotide linkages that are not natural phosphate linkages or their salt forms. Modified internucleotide linkages may also exist in salt forms depending on the structure. For example, as understood by those skilled in the art, phosphorothioate internucleotide linkages having the structure -OP(O)(SH)O- may exist in various salt forms (e.g., at physiological pH (about 7.4), the anion is -OP(O)(S - )O-Im).

[0388] In some embodiments, the oligonucleotide comprises a modified nucleotide linkage, e.g., a phosphorothioate, a phosphorodithioate, a methylphosphonate, a phosphoroamidate, a thiophosphate, a 3'-thiophosphate, or a 5'-thiophosphate. In some embodiments, the modified nucleotide linkage is a PN linkage. In some embodiments, the modified nucleotide linkage is a PS linkage. In some embodiments, the modified nucleotide linkage is a PO linkage (e.g., one other than a natural phosphate linkage). In some embodiments, each modified nucleotide linkage is independently a PN nucleotide linkage or a PS nucleotide linkage. In some embodiments, the oligonucleotide comprises one or more PN nucleotide linkages, one or more PS nucleotide linkages, and one or more PO nucleotide linkages. In some embodiments, one or more PN nucleotide connections are independently phosphoryl guanidine nucleotide connections. In some embodiments, one or more PN nucleotide connections are independently n001. In some embodiments, one or more PS nucleotide connections are independently phosphorothioate nucleotide connections. In some embodiments, each PS nucleotide connection is independently a phosphorothioate nucleotide connection. In some embodiments, one or more PO nucleotide connections are independently natural phosphate connections. In some embodiments, each PO nucleotide connection is independently a natural phosphate connection.

[0389] In some embodiments, the modified internucleotide linkage is a chiral internucleotide linkage containing a chiral linking factor. In some embodiments, the chiral internucleotide linkage is a phosphorothioate linkage. In some embodiments, the chiral internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, the chiral internucleotide linkage is a neutral internucleotide linkage. In some embodiments, the chiral internucleotide linkage is chirally controlled with respect to the chiral linking factor. In some embodiments, the chiral internucleotide linkage is stereochemically pure with respect to the chiral linking factor. In some embodiments, the chiral internucleotide linkage is not chirally controlled. In some embodiments, the backbone chiral center pattern is a chirally controlled internucleotide linkage ( R p or S It includes or consists of the position of p) and the position of the achiral nucleotide linkage (e.g., natural phosphate linkage).

[0390] In some embodiments, the internucleotide linkage includes a P-modification, and the P-modification is a modification in the linkage nucleolus. In some embodiments, the modified internucleotide linkage does not include a nucleolus but is a moiety that serves to link two sugars or two moietys each containing a nucleobase independently, as in, for example, peptide nucleic acid (PNA).

[0391] In some embodiments, internucleotide linkages are described in US 10167309, US 11643657, US 11718638, US 11608355, US 20230089442, US 20220306573 or US 20230203087, each of which is incorporated herein by reference.

[0392] In some embodiments, the modified internucleotide linkage (e.g., a non-negatively charged internucleotide linkage) comprises an optionally substituted triazolyl. In some embodiments, the modified internucleotide linkage (e.g., a non-negatively charged internucleotide linkage) comprises an optionally substituted alkynyl. In some embodiments, the modified internucleotide linkage comprises a triazole or alkyne moiety. In some embodiments, the triazole moiety (e.g., a triazolyl group) is optionally substituted. In some embodiments, the triazole moiety (e.g., a triazolyl group) is substituted. In some embodiments, the triazole moiety is not substituted. In some embodiments, the modified internucleotide linkage comprises an optionally substituted cyclic guanidine moiety. In some embodiments, the modified internucleotide linkage is It has the structure of and is selectively chiral controlled, where R 1 is -L-R', where each L and R' are independently as described herein. In some embodiments, each R 1 is independently R'. In some embodiments, each R' is independently R. In some embodiments, two R's 1 It is R and together forms a ring as described herein. In some embodiments, two Rs on two different nitrogen atoms 1 is R and together forms a ring as described herein. In some embodiments, R 1 C independently and optionally substituted as described herein 1-6 It is aliphatic. In some embodiments, R 1 It is methyl. In some embodiments, two R' on the same nitrogen atom are R and together form a ring as described herein. In some embodiments, the modified internucleotide linkage is It has the structure of and is optionally chiral controlled. In some embodiments, Is is. In some embodiments, the modified internucleotide linkage includes an optionally substituted cyclic guanidine moiety. , , or It has the structure, where W is O or S. In some embodiments, W is O. In some embodiments, W is S. In some embodiments, the linkage between non-negatively charged nucleotides is stereochemically controlled.

[0393] In some embodiments, the non-negatively charged nucleotide linkage or neutral nucleotide linkage is a nucleotide linkage comprising a triazole moiety. In some embodiments, the non-negatively charged nucleotide linkage or non-negatively charged nucleotide linkage comprises an optionally substituted triazolyl group. In some embodiments, the nucleotide linkage comprising a triazole moiety (e.g., an optionally substituted triazolyl group) has the following structure: . In some embodiments, the internucleotide linkage comprising a triazole moiety has the following structure: In some embodiments, internucleotide linkages comprising a triazole moiety are It has the chemical formula, where W is O or S. In some embodiments, the internucleotide linkage comprising an alkyne moiety (e.g., an optionally substituted alkynyl group) is It has the chemical formula, where W is O or S. In some embodiments, internucleotide linkages, e.g., non-negatively charged internucleotide linkages, neutral internucleotide linkages, comprise a cyclic guanidine moiety. In some embodiments, internucleotide linkages comprising a cyclic guanidine moiety have the following structure: . In some embodiments, negatively charged internucleotide connections, or neutral internucleotide connections, are , , , or It is a structure selected from or includes the same, where W is O or S.

[0394] In some embodiments, the linkage between nucleotides is a Tmg group ( Includes ). In some embodiments, the internucleotide linkage includes a Tmg group, and It has the structure of ("Tmg nucleotide linkage"). In some embodiments, neutral nucleotide linkages include nucleotide linkages of PNA and PMO, and Tmg nucleotide linkages.

[0395] In some embodiments, the negatively charged internucleotide linkage comprises an optionally substituted 3 to 20-membered heterocyclyl or heteroaryl group having 1 to 10 heteroatoms. In some embodiments, the negatively charged internucleotide linkage comprises an optionally substituted 3 to 20-membered heterocyclyl or heteroaryl group having 1 to 10 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, such heterocyclyl or heteroaryl group is of a 5-membered ring. In some embodiments, such heterocyclyl or heteroaryl group is of a 6-membered ring.

[0396] In some embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5- to 20-membered heteroaryl group having 1 to 10 heteroatoms. In some embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5- to 20-membered heteroaryl group having 1 to 10 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5- to 6-membered heteroaryl group having 1 to 4 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted 5-membered heteroaryl group having 1 to 4 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, the heteroaryl group is directly bonded to the linkage phosphorus. In some embodiments, the non-negatively charged internucleotide linkage comprises an optionally substituted triazolyl group. In some embodiments, the non-negatively charged internucleotide linkage comprises an unsubstituted triazolyl group, e.g. Includes. In some embodiments, the non-negatively charged internucleotide linkage is a substituted triazolyl group, e.g. Includes

[0397] In some embodiments, the negatively charged internucleotide linkage comprises an optionally substituted 5- to 20-membered heterocyclyl group having 1 to 10 heteroatoms. In some embodiments, the negatively charged internucleotide linkage comprises an optionally substituted 5- to 20-membered heterocyclyl group having 1 to 10 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, the negatively charged internucleotide linkage comprises an optionally substituted 5- to 6-membered heterocyclyl group having 1 to 4 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, the negatively charged internucleotide linkage comprises an optionally substituted 5-membered heterocyclyl group having 1 to 4 heteroatoms, wherein at least one heteroatom is nitrogen. In some embodiments, at least two heteroatoms are nitrogen. In some embodiments, the heterocyclyl group is directly bound to the linking phosphate. In some embodiments, the heterocyclyl group is bound to the linking phosphate via the linker, for example, if the heterocyclyl group is part of a guanidine moiety directly bound to the linking phosphate via =N-. In some embodiments, the non-negatively charged internucleotide linkage is optionally substituted It includes a group. In some embodiments, the non-negatively charged internucleotide linkage is substituted It includes a group. In some embodiments, the non-negatively charged internucleotide linkage is Includes the group, and each R 1 is independently -LR. In some implementations, each R 1 is independently and optionally substituted C 1-6 It is an alkyl. In some embodiments, each R 1 is independently methyl.

[0398] In some embodiments, the modified internucleotide linkage, e.g., the non-negatively charged internucleotide linkage, comprises a triazole or alkyne moiety, each of which is optionally substituted. In some embodiments, the modified internucleotide linkage comprises a triazole moiety. In some embodiments, the modified internucleotide linkage comprises an unsubstituted triazole moiety. In some embodiments, the modified internucleotide linkage comprises a substituted triazole moiety. In some embodiments, the modified internucleotide linkage comprises an alkyl moiety. In some embodiments, the modified internucleotide linkage comprises an optionally substituted alkynyl group. In some embodiments, the modified internucleotide linkage comprises an unsubstituted alkynyl group. In some embodiments, the modified internucleotide linkage comprises a substituted alkynyl group. In some embodiments, the alkynyl group is directly bound to the linkage phosphate.

[0399] In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, the non-negatively charged internucleotide linkage is It has the structure of. In some embodiments, W is O. In some embodiments, W is S. In some embodiments, the neutral nucleotide linkage is the aforementioned negatively uncharged nucleotide linkage.

[0400] In some embodiments, the oligonucleotide comprises neutral nucleotide-links and chiral-controlled nucleotide-links. In some embodiments, the oligonucleotide comprises neutral nucleotide-links and chiral-controlled nucleotide-links that are not neutral nucleotide-links. In some embodiments, the oligonucleotide comprises neutral nucleotide-links and chiral-controlled phosphorothioate nucleotide-links. In some embodiments, the present disclosure provides an oligonucleotide comprising one or more non-negatively charged nucleotide-links and one or more phosphorothioate nucleotide-links, wherein each phosphorothioate nucleotide-link in the oligonucleotide is independently a chiral-controlled nucleotide-link. In some embodiments, the present disclosure provides an oligonucleotide comprising one or more neutral nucleotide-links and one or more phosphorothioate nucleotide-links, wherein each phosphorothioate nucleotide-link in the oligonucleotide is independently a chiral-controlled nucleotide-link. In some embodiments, the oligonucleotide comprises at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more chiral-controlled phosphorothioate nucleotide-links. In some embodiments, the non-negatively charged nucleotide-links are chiral-controlled. In some embodiments, the non-negatively charged nucleotide-links are not chiral-controlled. In some embodiments, the neutral nucleotide-links are chiral-controlled. In some embodiments, the neutral nucleotide-links are not chiral-controlled. In some embodiments, the oligonucleotide comprises a link between one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) chiral-controlled and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) achiral-controlled chiral nucleotides.In some embodiments, the oligonucleotide comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) chiral-controlled and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) achiral-controlled negatively uncharged nucleotide linkages (in some embodiments, each of which is independently n001). In some embodiments, the neutral nucleotide linkages are chiral-controlled. In some embodiments, the neutral nucleotide linkages are not chiral-controlled. In some embodiments, the oligonucleotide comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) chiral-controlled and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) anachiral-controlled neutral nucleotide linkages (in some embodiments, each of these is independently n001).

[0401] In some embodiments, the internucleotide linkage has the structure -OP(=W)[-N(R')2]-O-, where W is O or S, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -OP(=W)(-NHR')-O-, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -OP(=W)(-NHSO2R)-O-, and each variable is as independently described herein. In some embodiments, R is methyl. In some embodiments, the internucleotide linkage is -OP(=O)(-NHSO2CH3)-O-. In some embodiments, the internucleotide linkage has the structure -OP(=W)[-N=C(-R')2]-O-, and each variable is as independently described herein. In some embodiments, the nucleotide linkage has the structure -OP(=W)[-N=C[N(R')2]2]-O-, and each variable is as independently described herein. In some embodiments, the nucleotide linkage has the structure -OP(=W)(-N=C(R')2)-O-, and each variable is as independently described herein. In some embodiments, the nucleotide linkage has the structure -OP(=W)(-N(R')2)-O-, and each variable is as independently described herein. In some embodiments, W is O. In some embodiments, W is S. In some embodiments, this nucleotide linkage is a non-negatively charged nucleotide linkage. In some embodiments, this nucleotide linkage is a neutral nucleotide linkage.

[0402] In some embodiments, the internucleotide linkage has the structure -P(=W)[-N(R')2]-O-, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)(-NHR')-O-, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)(-NHSO2R)-O-, and each variable is as independently described herein. In some embodiments, R is methyl. In some embodiments, the internucleotide linkage is -P(=O)(-NHSO2CH3)-O-. In some embodiments, the internucleotide linkage is -P(=W)[-N=C(-L L It has the structure -R')2]-O-, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)[-N=C[N(R')2]2]-O-, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)(-N=C(R')2)-O-, and each variable is as independently described herein. In some embodiments, the internucleotide linkage has the structure -P(=W)(-N(R')2)-O-, and each variable is as independently described herein. In some embodiments, W is O. In some embodiments, W is S. In some embodiments, this internucleotide linkage is a non-negatively charged internucleotide linkage. In some embodiments, this internucleotide linkage is a neutral internucleotide linkage. In some embodiments, the P of this internucleotide linkage is bound to the N of the sugar.

[0403] In some embodiments, the linkage is between phosphoryl guanidine nucleotides. In some embodiments, the linkage is between thiophosphoryl guanidine nucleotides.

[0404] Oligonucleotide

[0405] In some embodiments, the provided technology is useful for preparing or delivering oligonucleotides comprising -P(O)(OH)2 which may exist in various salt forms. In some embodiments, the present disclosure is as described herein -P(O)(OR PG Provides an oligonucleotide comprising )2. In some embodiments, -P(O)(OR as described herein PG Oligonucleotides containing )2 are administered to a subject to prevent or treat a pathological condition, disorder, or disease. In some embodiments, -P(O)(OR as described herein PG Oligonucleotides containing -P(O)(OH)2 are utilized to prepare oligonucleotides containing -P(O)(OH)2, which may exist in various salt forms. For example, in some embodiments, the present disclosure is Provides a compound that is an oligonucleotide containing, wherein R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is optionally substituted tert-butyl. In some embodiments, the present disclosure provides a compound that is an oligonucleotide comprising a moiety of formula O or a salt thereof:

[0406] ,

[0407] O

[0408] Each variable herein is as independently described herein. In some embodiments, the present disclosure provides a compound that is an oligonucleotide comprising a moiety of the formula OI or a salt thereof:

[0409] ,

[0410] OI

[0411] Each variable herein is as independently described herein. In some embodiments, the present disclosure provides a compound that is an oligonucleotide comprising a moiety of the formula O-I' or a salt thereof:

[0412] ,

[0413] OI'

[0414] Here, each variable is independently as described herein.

[0415] As recognized by those skilled in the art, oligonucleotides may comprise various sugars, nucleobases, and nucleotide-linkages as described herein. In some embodiments, the oligonucleotide comprises PS linkages. In some embodiments, the oligonucleotide comprises PN linkages. In some embodiments, the oligonucleotide comprises PO linkages. In some embodiments, the oligonucleotide comprises PS linkages and PN linkages. In some embodiments, the oligonucleotide comprises PS linkages and PO linkages. In some embodiments, the oligonucleotide comprises PO linkages, PS linkages, and PN linkages. In some embodiments, the number of PS nucleotide-linkages within the oligonucleotide is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more. In some embodiments, the number of PN nucleotide linkages of the oligonucleotide is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more. In some embodiments, the number of PO nucleotide linkages of the oligonucleotide is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more. In some embodiments, each PS linkage is a linkage between phosphorothioate nucleotides. In some embodiments, each PN linkage is independently a linkage between non-negatively charged nucleotides. In some embodiments, each PN link is independently a link between neutral nucleotides. In some embodiments, each PN link is independently a link between phosphoryl guanidine nucleotides. In some embodiments, each PN link is independently n001.

[0416] In some embodiments, the linkage between each PS nucleotide is independently chiral controlled. In some embodiments, the linkage between each PN nucleotide is independently chiral controlled. In some embodiments, each chiral linkage is independently chiral controlled.

[0417] In some embodiments, the oligonucleotides are about 10 to 200 (e.g., about 10 to 20, 10 to 30, 10 to 40, 10 to 50, 10 to 60, 10 to 70, 10 to 80, 10 to 90, 10 to 100, 10 to 120, 10 to 150, 20 to 30, 20 to 40, 20 to 50, 20 to 60, 20 to 70, 20 to 80, 20 to 90, 20 to 100, 20 to 120, 20 to 150, 20 to 200, 25 to 30, 25 to 40, 25 to 50, 25 to 60, 25 to 70, 25 to 80, 25 to 90, 25 to 100, It has a nucleotide length of 25~120, 25~150, 25~200, 30~40, 30~50, 30~60, 30~70, 30~80, 30~90, 30~100, 30~120, 30~150, 30~200, 10, 20, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 45, 50, 60, etc.). In some embodiments, the nucleotide sequence of the oligonucleotide is about 10~60 nucleotides long. In some embodiments, the nucleotide sequence is about 15~50 nucleotides long. In some embodiments, the base sequence is about 15 to about 35 nucleotides long. In some embodiments, the oligonucleotide is about 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 or more nucleotides long. In some embodiments, each nucleobase calculated by length is an independently optionally substituted or protected A, T, C, G, U, or a tautomer thereof.

[0418] In some embodiments, the oligonucleotide is provided in the form of a salt. In some embodiments, the oligonucleotide is provided as a salt comprising negatively charged internucleotide links (e.g., phosphorothioate internucleotide links, natural phosphate links, etc.) present in its salt form. In some embodiments, the oligonucleotide is provided as a pharmaceutically acceptable salt. In some embodiments, the oligonucleotide is provided as a metal salt. In some embodiments, the oligonucleotide is provided as a sodium salt. In some embodiments, the oligonucleotide is provided as an ammonium salt. In some embodiments, the oligonucleotide is provided as a metal salt, e.g., a sodium salt, and each negatively charged internucleotide link is independently in the form of a salt (e.g., in the sodium salt, -OP(O)(SNa)-O- for phosphorothioate internucleotide links, -OP(O)(ONa)-O- for natural phosphate links, etc.).

[0419] In some embodiments, the oligonucleotide comprises one or more chiral-controlled nucleotide linkages and is chiral-controlled. In some embodiments, the provided oligonucleotide is stereochemically pure. In some embodiments, the provided oligonucleotide or its composition is substantially free of other stereoisomers. In some embodiments, the present disclosure provides a chiral-controlled oligonucleotide composition.

[0420] As described herein, the oligonucleotides of the present disclosure may be provided in high purity (e.g., 50% to 100%). In some embodiments, the oligonucleotides of the present disclosure have high stereochemical purity (e.g., 50% to 100%). In some embodiments, the oligonucleotides in the provided composition have high stereochemical purity (e.g., a high proportion (e.g., 50% to 100%) of stereoisomers compared to other stereoisomers of the same oligonucleotide). In some embodiments, the percentage is at least or about 50%. In some embodiments, the percentage is at least or about 60%. In some embodiments, the percentage is at least or about 70%. In some embodiments, the percentage is at least or about 75%. In some embodiments, the percentage is at least or about 80%. In some embodiments, the percentage is at least or about 85%. In some embodiments, the percentage is at least or about 90%. In some implementations, the percentage is at least or about 95%.

[0421] In some embodiments, the oligonucleotides of the present disclosure are stereochemically pure at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% in the linkage of the chiral nucleotide linkages. In some embodiments, the oligonucleotides of the present disclosure are stereoselectively prepared and are substantially free of stereoisomers. In some embodiments, the provided composition (e.g., comprising a plurality of oligonucleotides sharing the same base sequence of the same chiral linkage stereochemical pattern) R p and / or S It includes one or more of p, and each chiral linkage factor independently R p or SIn p), at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of all oligonucleotides in the composition sharing the same base sequence as the plurality of oligonucleotides share the same chiral linkage stereochemical pattern or are multiple oligonucleotides. In some embodiments, in the provided composition comprising multiple oligonucleotides sharing the same base sequence of the same chiral linkage stereochemical pattern, at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of all oligonucleotides in the composition sharing the same composition as the plurality of oligonucleotides share the same chiral linkage stereochemical pattern or are multiple oligonucleotides. In some embodiments, the excess of the diastereomer of each chiral is independently about or at least about 90%. In some embodiments, the diastereomer excess of each chiral phosphate is independently about or at least about 95%. In some embodiments, the diastereomer excess of each chiral phosphate is independently about or at least about 97%. In some embodiments, the diastereomer excess of each chiral phosphate is independently about or at least about 98%. In some embodiments, the diastereomer purity is about or at least about (DS). nc And, where DS is about 90% to 100%, and nc is the number of chiral linked phosphate groups. In some embodiments, DS is about 90% or more. In some embodiments, DS is about 95% or more. In some embodiments, DS is about 96% or more. In some embodiments, DS is about 97% or more. In some embodiments, DS is about 98% or more. In some embodiments, DS is about 99% or more. In some embodiments, DS is about 99% or more. In some embodiments, the diastereomer purity is expressed as the product of the diasterepurities of each chiral linked phosphate group.

[0422] Oligonucleotide composition

[0423] In particular, the present invention provides various oligonucleotide compositions. In some embodiments, the present invention provides an oligonucleotide composition of the oligonucleotides described herein. In some embodiments, the oligonucleotide composition comprises a plurality of oligonucleotides described in the present invention. In some embodiments, the oligonucleotide composition is chirally controlled. In some embodiments, the oligonucleotide composition is not chirally controlled (stereo-random).

[0424] The linking factor of natural phosphate linkages is chiral. The linking factor of many modified nucleotide linkages, e.g., phosphorothioate nucleotide linkages, is chiral. In some embodiments, during the preparation of an oligonucleotide composition (e.g., in traditional phosphoramidite oligonucleotide synthesis), the arrangement of chiral linking factors is not intentionally designed or controlled, resulting in a non-chiral controlled (stereo-random) oligonucleotide composition (substantially racemic) which is a complex and random mixture of various stereoisomers (diastereo-isomers), and for an oligonucleotide having n chiral nucleotide linkages (linking factors are chiral), there are generally 2^n stereoisomers (e.g., when n is 10, 2^10 = 1,032; when n is 20, 2^20 = 1,048,576). These stereoisomers have the same composition but differ in the stereochemical pattern of the linking phosphate.

[0425] In some embodiments, stereorandom oligonucleotide compositions have properties and / or activities sufficient for a specific purpose and / or application. In some embodiments, the preparation of stereorandom oligonucleotide compositions may be cheaper and / or easier and simpler than that of chiral-controlled oligonucleotide compositions. However, stereoisomers within stereorandom compositions may have different properties, activities, and / or toxicity, which may result in inconsistent therapeutic effects and / or unintended side effects caused by stereorandom compositions, particularly compared to specific chiral-controlled oligonucleotide compositions of the same composition.

[0426] In some embodiments, the present invention includes techniques for designing and manufacturing chiral-controlled oligonucleotide compositions. In some embodiments, the chiral-controlled oligonucleotide composition comprises a plurality of oligonucleotides at a controlled / predetermined level (not random as in stereorandom compositions), and the oligonucleotides share the same linkage stereochemistry in one or more chiral nucleotide-linkages (chiral-controlled nucleotide-linkages). In some embodiments, the oligonucleotides share the same backbone chiral center pattern (stereochemistry of the linkage). In some embodiments, the backbone chiral center pattern is as described in the present invention. In some embodiments, the plurality of oligonucleotides are structurally identical.

[0427] In some embodiments, the present invention provides a chiral-controlled oligonucleotide composition comprising a plurality of oligonucleotides, wherein the oligonucleotides are:

[0428] 1) Common configuration and

[0429] 2) sharing the same stereochemistry in one or more chiral nucleotide linkages (chiral-controlled nucleotide linkages), (e.g., 1–50, 1–40, 1–30, 1–25, 1–20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more), and

[0430] The composition is rich in multiple oligonucleotides compared to a substantially racemic formulation of oligonucleotides of common composition.

[0431] In some embodiments, the present invention provides a chiral-controlled oligonucleotide composition comprising a plurality of oligonucleotides, wherein the oligonucleotides are:

[0432] 1) Common configuration and

[0433] 2) sharing the same stereochemistry in one or more chiral nucleotide linkages (chiral-controlled nucleotide linkages), (e.g., 1–50, 1–40, 1–30, 1–25, 1–20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more), and

[0434] The stereochemical purity of the linkage of each chiral-controlled internucleotide linkage is independently 80% to 100% (e.g., 85% to 100%, 90% to 100%, about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5%).

[0435] In some embodiments, at least one linkage between chiral nucleotides is chiral controlled. In some embodiments, at least two linkages between nucleotides are independently chiral controlled. In some embodiments, the number of chiral-controlled linkages between nucleotides is at least 3. In some embodiments, the number is at least 4. In some embodiments, the number is at least 5. In some embodiments, the number is at least 6. In some embodiments, the number is at least 7. In some embodiments, the number is at least 8. In some embodiments, the number is at least 9. In some embodiments, the number is at least 10. In some embodiments, the number is at least 11. In some embodiments, the number is at least 12. In some embodiments, the number is at least 13. In some embodiments, the number is at least 14. In some embodiments, the number is at least 15. In some embodiments, the number is at least 20. In some embodiments, the number is at least 25. In some embodiments, the number is at least 30.

[0436] In some embodiments, at least 5% to 100% of all chiral linkages (e.g., about 10% to 100%, 20% to 100%, 30% to 100%, 40% to 100%, 50% to 80%, 50% to 85%, 50% to 90%, 50% to 95%, 60% to 80%, 60% to 85%, 60% to 90%, 60% to 95%, 60% to 100%, 65% to 80%, 65% to 85%, 65% to 90%, 65% to 95%, 65% to 100%, 70% to 80%, 70% to 85%, 70% to 90%, 70% to 95%, 70% to 100%, 75%~80%, 75%~85%, 75%~90%, 75%~95%, 75%~100%, 80%~85%, 80%~90%, 80%~95%, 80%~100%, 85%~90%, 85%~95%, 85%~100%, 90%~95%, 90%~100%, 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%, etc.) are chiral controlled. In some embodiments, at least 5% to 100% of the links between all phosphorothioate nucleotide...

Claims

Claim 1 As a compound, the compound is a compound of formula I' or a salt thereof: ,I'here:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl; SU is a sugar; BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase; and R 1 , R 2 and R 3 Each is independently R' or R 1 , R 2 and R 3 Two or three of them are taken together with the atoms interposed between them It forms; ring A is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms; t is 0-5; and each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R s11 , -L s -OR s11 , -L s -SR s11 , -L s -N(R s11 )2, -OL s -OR s11 , -OL s -SR s11 , -OL s -N(R s11 )2, -C(R s11 )3 or -L s -Si(R s11 )3 and; each R s11 is independently R' and each L s is independently L; each L is independently covalently bonded, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-; -Cy- is an optionally substituted divalent 3-20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' is taken together with the interposed atom(s) to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s); and each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 A compound having a 5- to 14-membered heteroaryl ring having 1- to 5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3- to 15-membered heterocyclic ring having 1- to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, selected as an optionally substituted group. Claim 2 As a compound, the compound is a compound of formula I or a salt thereof: ,I here:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl; BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase; and R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a , -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s and; each L 2s is independently L; each R 2a is independently R'; each L is independently a covalent bond, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-; -Cy- is an optionally substituted divalent 3- to 20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and R 1 , R 2 and R 3 Each is independently R' or R 1 , R 2 and R 3 Two or three of them are taken together with the atoms interposed between them It forms; ring A is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms; t is 0-5; and each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R s11 , -L s -OR s11 , -L s -SR s11 , -L s -N(R s11 )2, -OL s -OR s11 , -OL s -SR s11 , -OL s -N(R s11 )2, -C(R s11 )3 or -L s -Si(R s11 )3 and; each R s11 is independently R' and each L s is independently L; each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' are taken together with an intervening atom(s) to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the intervening atom(s); and each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 A compound having a 5- to 14-membered heteroaryl ring having 1- to 5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3- to 15-membered heterocyclic ring having 1- to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, selected as an optionally substituted group. Claim 3 In any one of paragraphs 1 to 2, R 1 A compound that is -CH2CH2CN. Claim 4 In any one of paragraphs 1 to 3, R 2 is optionally substituted C 1-6 A compound that is aliphatic (e.g., isopropyl). Claim 5 In any one of paragraphs 1 through 4, R 3 is optionally substituted C 1-6 A compound that is aliphatic (e.g., isopropyl). Claim 6 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 ) is a compound that is -P(OCH2CH2CN)N[CH(CH3)2]2. Claim 7 In paragraph 1, R 1 , R 2 and R 3 Two or three of them are taken together with the atoms interposed between them A compound that forms. Claim 8 In paragraph 7, the compound is a polycyclic ring in which ring A is an optionally substituted ring. Claim 9 In paragraph 7, the compound is a ring A being an optionally substituted cyclic 7- to 12-membered ring. Claim 10 In Paragraph 7, ring A is -P(OR 1 )N(R 2 )(R 3 A compound comprising an optionally substituted 5- or 6-membered ring containing phosphorus, oxygen, and nitrogen atoms. Claim 11 In Clause 10, -P(OR 1 )N(R 2 )(R 3 A compound in which the ring containing phosphorus, oxygen, and nitrogen atoms does not have other ring heteroatoms. Claim 12 A compound according to claim 7, wherein each monocyclic ring unit of ring A is an independently and optionally substituted 3- to 7-membered ring. Claim 13 In paragraph 7, ring A is optionally substituted or including this, where each L a is independently a covalent bond or divalent C 1-5 It is an aliphatic group, and each L a A compound in which one or more methylene units are optionally and independently replaced with -O-, -S-, or -NH. Claim 14 In paragraph 7, ring A is optionally substituted A compound that is or contains the same. Claim 15 In paragraph 7, ring A is optionally substituted A compound that is or contains the same. Claim 16 In paragraph 7, ring A is optionally substituted A compound that is or contains the same. Claim 17 A compound in any one of paragraphs 7 to 16, wherein t is 1 to 5. Claim 18 In any one of paragraphs 7 through 16, R s The existence of -P(OR 1 )N(R 2 )(R 3 A compound that is bonded to a carbon atom bonded to the oxygen of ). Claim 19 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )Is and, here R s1 and R s2 Each independently R s Phosphorus, compound. Claim 20 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver Phosphorus, compound. Claim 21 In any one of paragraphs 7 through 20, L s The presence of is a compound that is selectively substituted -CH2-. Claim 22 In any one of paragraphs 7 through 20, L s A compound in which the presence of is a covalent bond. Claim 23 In any one of paragraphs 7 through 22, R s The existence of -L s -Si(R s11 )3-person, compound. Claim 24 In Paragraph 23, each R s11 It is a compound that is not hydrogen. Claim 25 In Paragraph 23, each R s11 C independently 1-6 A compound selectively substituted from aliphatic and phenyl compounds. Claim 26 In Paragraph 23, -Si(R s11 )3 is a compound that is -Si(CH3)(Ph)2. Claim 27 In any one of paragraphs 7 through 22, R s -L s -R s11 Phosphorus, compound. Claim 28 In Paragraph 27, R s11 is -S(O)2R, where R is an optionally substituted C 1-6 Aliphatic, compound. Claim 29 In Paragraph 27, R s11 A compound of -S(O)2R, where R is t-Bu. Claim 30 In Paragraph 27, R s11 is -S(O)2R, where R is methyl, a compound. Claim 31 In Paragraph 27, R s11 A compound that is -S(O)2R. Claim 32 In any one of paragraphs 7 through 20, R s The existence of -C(R s11 )3-person, compound. Claim 33 In any one of paragraphs 7 through 20, R s The existence of -CH(R s11 )2-person, compound. Claim 34 In paragraph 33, 2 Rs s11 A compound that takes together with the carbon atoms to which they are attached to form an optionally substituted 3 to 20-membered ring having 0 to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. Claim 35 In paragraph 20, R s2 is optionally substituted C 1-6 Aliphatic, or R s2 is a compound that is an optionally substituted phenyl. Claim 36 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver Phosphorus, compound. Claim 37 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver or Phosphorus, compound. Claim 38 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver Phosphorus, compound. Claim 39 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver or Phosphorus, compound. Claim 40 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver , or Phosphorus, compound. Claim 41 In paragraph 1, -P(OR 1 )N(R 2 )(R 3 )silver , , , , or Phosphorus, compound. Claim 42 As a compound, the above compound is It is an oligonucleotide containing, where R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is a compound that is optionally substituted tert-butyl. Claim 43 As a compound, the compound is an oligonucleotide comprising a moiety of the formula OI or a salt thereof: ,OI Here:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl; BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase; and R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a , -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s and; each L 2s is independently L; each R 2a is independently R' and each L is independently C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-; -Cy- is an optionally substituted divalent 3-20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' is taken together with the interposed atom(s) to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s); and each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 A compound having a 5- to 14-membered heteroaryl ring having 1- to 5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3- to 15-membered heterocyclic ring having 1- to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, selected as an optionally substituted group. Claim 44 As a compound, the compound is a compound of formula II' or a salt thereof: ,II'here:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P A compound in which is an optionally substituted tert-butyl; SU is a sugar; and BA is an optionally substituted 3- to 20-membered ring having 0 to 10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase. Claim 45 As a compound, the compound is a compound of Formula II or a salt thereof: ,IIhere:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl; BA is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is an optionally substituted or protected nucleobase; and R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a , -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s and; each L 2s is independently L; each R 2a is independently R' and each L is independently C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-; -Cy- is an optionally substituted divalent 3-20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' is taken together with the interposed atom(s) to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s); and each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 A compound having a 5- to 14-membered heteroaryl ring having 1- to 5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3- to 15-membered heterocyclic ring having 1- to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, selected as an optionally substituted group. Claim 46 In any one of paragraphs 1 through 45, R P1 and R P2 is a compound that is -O-CH2CH2CN. Claim 47 In any one of paragraphs 1 through 46, R P1 and R P2 is a compound that is -O-CH2-OC(O)-tBu. Claim 48 As a compound, the compound is a compound of formula III or formula III' or a salt thereof: ,III ,III' where: BA is a selectively substituted 3–20-membered ring having 0–10 heteroatoms independently selected from hydrogen, or silicon, nitrogen, oxygen, phosphorus, and sulfur; or BA is a selectively substituted or protected nucleobase; SU is a sugar; and R 2s -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L 2s -R 2a , -L 2s -OR 2a , -L 2s -SR 2a , -L 2s -N(R 2a )2, -OL 2s -OR 2a , -OL 2s -SR 2a or -OL 2s -N(R 2a )2 and, or R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s and; each L 2s is independently L; each R 2a is independently R' and each L is independently C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-; -Cy- is an optionally substituted divalent 3-20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' is taken together with the interposed atom(s) to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s); and each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 A compound having a 5- to 14-membered heteroaryl ring having 1- to 5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3- to 15-membered heterocyclic ring having 1- to 5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, selected as an optionally substituted group. Claim 49 A compound according to any one of claims 1 to 48, wherein BA is an optionally substituted 3- to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. Claim 50 In any one of claims 1 to 49, BA is a compound bonded to C1 at a nitrogen atom. Claim 51 In any one of claims 1 to 50, BA is an optionally substituted group, wherein the group , , , , or or a compound that is a tautomeric isomer thereof. Claim 52 A compound according to any one of claims 1 to 51, wherein BA is an optionally protected nucleobase. Claim 53 In paragraph 52, the above-mentioned nucleobase is a compound selected from A, T, C, G, U, and 5mU. Claim 54 In any one of paragraphs 1 through 53, R 2s is a compound that is -H. Claim 55 In any one of paragraphs 1 through 53, R 2s is a compound that is -F. Claim 56 In any one of paragraphs 1 through 53, R 2s is -OR 2a Phosphorus, compound. Claim 57 In Paragraph 56, R 2a is a methyl compound. Claim 58 In any one of paragraphs 1 through 53, L 2s is a compound that is not a covalent bond. Claim 59 In any one of paragraphs 1 through 53, L 2s is optionally substituted C 1-10 Alkylene, compound. Claim 60 In any one of paragraphs 1 through 53, R 2s is -L 2s -R 2a Phosphorus, compound. Claim 61 In Paragraph 60, L 2s is -OL 2s' -R 2a and, here L 2s' is an optionally substituted -(CH2) 1-10 - Phosphorus, compound. Claim 62 In any one of paragraphs 1 through 53, R 2s is -OL 2s -OR 2a Phosphorus, compound. Claim 63 In Paragraph 62, R 2a is a compound that is -OMe. Claim 64 In Paragraph 62, R 2s is a compound that is -OCH2CH2OMe. Claim 65 In any one of paragraphs 1 through 53, R 2s is L connecting C2 with C1, C2, C3, C4, or C5 2s Phosphorus, compound. Claim 66 A compound according to any one of claims 1 to 53 and 65, wherein L is (C2)-O-(unsubstituted methylene)-phosphorus. Claim 67 A compound according to any one of claims 1 to 53 and 65, wherein L is (C2)-O-(substituted methylene)-phosphorus. Claim 68 In paragraph 67, the carbon atom of the methylene group is R Phosphorus, compound. Claim 69 In paragraph 67, the carbon atom of the methylene group is S Phosphorus, compound. Claim 70 As a compound, the compound is a compound of formula IV or a salt thereof: ,IV Here:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is a compound that is optionally substituted tert-butyl. Claim 71 In Paragraph 70, R P1 and R P2 is a compound that is -O-CH2CH2CN. Claim 72 In Paragraph 70, R P1 and R P2 is a compound that is -O-CH2-OC(O)-tBu. Claim 73 A compound according to any one of claims 1 to 72, wherein the purity of the compound is about 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or higher. Claim 74 A method for preparing an oligonucleotide comprising the step of converting to -P(O)(OH)2 (e.g., its salt form). Claim 75 A method for producing an oligonucleotide comprising the step of coupling a compound of any one of claims 1 to 41, 45 to 47 and 49 to 69, which is a compound of formula I or formula I' or a salt thereof, to an oligonucleotide. Claim 76 A method for preparing an oligonucleotide comprising the step of reacting a compound of any one of claims 1 to 41, 45 to 47 and 49 to 69, which is a compound of formula I or formula I' or a salt thereof, with a compound containing an -OH group. Claim 77 In paragraph 76, the method wherein the -OH group is the 5'-OH of an oligonucleotide. Claim 78 A method according to any one of claims 75 to 77, wherein the preparation of the oligonucleotide comprises utilizing a solid support. Claim 79 A method comprising the step of preparing a compound of any one of claims 42, 46 to 47 and 49 to 69 in any one of claims 75 to 78, wherein the compound is a compound of the formula OI or a salt thereof. Claim 80 In any one of paragraphs 75 through 78, A method comprising the step of converting to -P(O)(OH)2 (e.g., in the salt form thereof). Claim 81 In Paragraph 80, R P1 and R P2 A method in which -OCH2CH2CN is used, and the conversion is performed in the presence of a base. Claim 82 In Paragraph 80, R P1 and R P2 is a method that is -OCH2OC(O)tBu. Claim 83 A method according to any one of claims 75 to 82, wherein the conversion occurs during cleavage and deprotection of removing a base protecting group and / or a chiral supporting group and / or cleaving an oligonucleotide from a solid support. Claim 84 A method according to any one of claims 75 to 83, wherein the oligonucleotide produced is a compound according to any one of claims 42, 46 to 47 and 49 to 69, wherein the compound is a compound of the formula OI or a salt thereof. Claim 85 A method for preparing a compound of any one of claims 1 to 41, 45 to 47, and 49 to 69, which is a compound of formula I or formula I' or a salt thereof, comprising the step of reacting a compound of any one of claims 45 to 47 and 49 to 69, which is a compound of formula II or formula II' or a salt thereof, with a compound of formula V or a salt thereof: ,V Here:LG is the departure phase and;R 1 , R 2 and R 3 Each is independently R' or R 1 , R 2 and R 3 Two or three of them are taken together with the atoms interposed between them It forms; ring A is an optionally substituted 3- to 20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atoms; t is 0-5; and each R s is independently -F, -Cl, -Br, -I, -CN, -N3, -NO, -NO2, -L s -R s11 , -L s -OR s11 , -L s -SR s11 , -L s -N(R s11 )2, -OL s -OR s11 , -OL s -SR s11 , -OL s -N(R s11 )2, -C(R s11 )3 or -L s -Si(R s11 )3 and; each R s11 is independently R' and each L s is independently L; each L is independently C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic and silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 A divalent optionally substituted group selected from heteroaliphatic groups, wherein one or more methylene units of L are optionally and independently replaced with -Cy-, -O-, -S-, -N(R')-, -C(O)-, -C(S)-, -C(NR')-, -C(O)N(R')-, -N(R')C(O)N(R')-, -N(R')C(O)O-, -S(O)-, -S(O)2-, -S(O)2N(R')-, -SC(O)- or -C(O)O-; -Cy- is an optionally substituted divalent 3-20-membered ring having 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R' is taken together with the interposed atom(s) to form an optionally substituted 3 to 20-membered ring having 0 to 10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur in addition to the interposed atom(s); and each R is independently hydrogen, or C 1-10 C having 1 to 5 heteroatoms independently selected from aliphatic, silicon, nitrogen, oxygen, phosphorus, and sulfur 1-10 Heterolipid, C 6-14 A method comprising a 5-14 member heteroaryl ring having 1-5 heteroatoms independently selected from aryl, silicon, nitrogen, oxygen, phosphorus, and sulfur, and a 3-15 member heterocyclic ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, with an optionally substituted group selected from the same. Claim 86 In paragraph 85, LG is -Cl, method. Claim 87 A method according to any one of paragraphs 85 to 86, wherein the reaction is performed in the presence of a base. Claim 88 In any one of paragraphs 85 to 87, R of formula V 1 , R 2 and R 3 The method as described in any one of paragraphs 3 through 41. Claim 89 A method comprising the method of any one of paragraphs 85 through 88 in any one of paragraphs 75 through 84. Claim 90 A method for preparing a compound of any one of claims 45 to 47 and 49 to 69, which is a compound of formula II or formula II' or a salt thereof, comprising the step of reacting a compound of any one of claims 48 to 69, which is a compound of formula III or formula III' or a salt thereof, with a compound of any one of claims 70 to 73. Claim 91 In paragraph 90, the above reaction is performed in the presence of a base. Claim 92 In paragraph 91, the method wherein the base is DIEA. Claim 93 A method according to any one of claims 90 to 92, wherein the reaction is carried out in the presence of a copper salt. Claim 94 A method according to any one of claims 90 to 92, wherein the reaction is carried out in the presence of CuI. Claim 95 A method comprising the method of any one of paragraphs 75 through 89, wherein the method of any one of paragraphs 90 through 94. Claim 96 A method for preparing a compound of any one of claims 70 to 73, comprising the step of oxidizing a compound of formula VI or a salt thereof: ,VI Here:R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl, method. Claim 97 In paragraph 96, the oxidizing agent is mCPBA, method. Claim 98 In paragraph 96 or 97, R P1 and R P2 is a method that is -O-CH2CH2CN. Claim 99 A method comprising the method of any one of paragraphs 75 to 95, and any one of paragraphs 96 to 98. Claim 100 As a method for preparing a compound of formula VI or a salt thereof: The method comprises the step of reacting VI≡-MgBr with a compound of formula VII or a salt thereof: ,VIIHere:LG is the departure phase; and R P1 and R P2 Each is -O-CH2CH2CN or -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl, method. Claim 101 In Clause 100, LG is -Cl, method. Claim 102 In any one of paragraphs 100 to 101, R P1 and R P2 Each is a method that is -O-CH2CH2CN. Claim 103 A method comprising the method of any one of paragraphs 100 to 102 in any one of paragraphs 75 to 99. Claim 104 As a method for preparing a compound of formula VII or a salt thereof: ,VII A method comprising the step of reacting HO-CH2CH2-CN with PCl3 in the presence of a base, wherein LG is -Cl. Claim 105 In paragraph 104, the above base is TEA, method. Claim 106 A method comprising the method of any one of paragraphs 75 to 103, wherein any one of paragraphs 104 to 105. Claim 107 A method for preparing a compound according to any one of claims 70 to 73, wherein R P1 and R P2 Each is -O-CH2-OC(O)-R P and, here R P is an optionally substituted tert-butyl, and a compound of formula IX or a salt thereof: , comprising the step of reacting IX with a compound having the structure of chemical formula X or a salt thereof:LG-CH2-OC(O)-R P , X here:R P is an optionally substituted tert-butyl; LG is a leaving group; and R P3 and R P4 Each independently -OR SP and, here R SP is optionally substituted C 1-6 Aliphatic, method. Claim 108 In Paragraph 107, R P3 and R P4 Each is a -OMe method. Claim 109 A method in which LG is -I in any one of paragraphs 107 to 108. Claim 110 A method comprising the method of any one of paragraphs 107 to 109 in any one of paragraphs 75 to 95. Claim 111 As a method for preparing a compound of chemical formula IX or a salt thereof: ,IX≡-MgBr comprises the step of reacting with a compound of formula XI or a salt thereof: ,XI Here: LG is the departure phase and R P3 and R P4 Each independently -OR SP and, here, each R SP is independently and optionally substituted C 1-6 Aliphatic, method. Claim 112 In Paragraph 111, R P3 and R P4 Each is a -OMe method. Claim 113 A method in which LG is -Cl in any one of paragraphs 111 to 112. Claim 114 A method comprising the method of any one of paragraphs 75 through 95 and paragraphs 107 through 110, wherein the method of any one of paragraphs 111 through 113. Claim 115 A method for preparing a compound of any one of claims 48 to 69 which is a compound of formula III or formula III' or a salt thereof, wherein the compound is of formula XII or formula XII' or a salt thereof: ,XII ,XII'where LG is a leaving group, a method comprising the step of reacting a compound or a salt thereof with an azide. Claim 116 In Clause 115, LG is -I, method. Claim 117 A method according to any one of claims 115 to 116, wherein the azide is sodium azide. Claim 118 A method comprising the method of any one of paragraphs 115 through 117 in any one of paragraphs 75 through 114. Claim 119 As a method for preparing a compound of formula XII or XII' or a salt thereof: ,XII ,XII'where LG is -I, and a compound of formula XIII or formula XIII' or a salt thereof: ,XIII A method comprising the step of reacting ,XIII' with I2. Claim 120 A method according to any one of claims 115 to 119, wherein the reaction is performed in the presence of phosphine. Claim 121 A method according to any one of claims 115 to 119, wherein the reaction is carried out in the presence of PPh3. Claim 122 A method according to any one of claims 119 to 121, wherein the reaction is performed in the presence of a base. Claim 123 A method according to any one of claims 119 to 121, wherein the reaction is carried out in the presence of a heteroaryl base, wherein the heteroaryl ring comprises -N= or -N(R)-. Claim 124 A method according to any one of claims 119 to 121, wherein the reaction is carried out in the presence of a heteroaryl base, wherein the heteroaryl ring comprises -N= or -NH-. Claim 125 A method according to any one of claims 119 to 121, wherein the reaction is carried out in the presence of imidazole. Claim 126 A method comprising the method of any one of paragraphs 119 through 125 in any one of paragraphs 75 through 118. Claim 127 A compound or method of any one of Embodiments 1 to 205.

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