Methods and reagents for synthesizing nucleoside analogs, and their applications.

Abstract

The present invention relates to a method for synthesizing nucleoside analogs. More specifically, the present invention relates to a method for synthesizing nucleoside analogs in which the C4' position is substituted.

Description

field

[0001] This invention relates to a method for synthesizing nucleoside analogs. More specifically, this invention relates to a method for synthesizing nucleoside analogs in which the C4' position is substituted. Background

[0002] Nucleosides play crucial roles in a variety of cellular processes, from cellular signaling to metabolism (1). Nucleosides consist of a nucleic acid base (typically adenine, guanine, cytosine, thymine, and uracil) and a sugar moiety (typically ribose or 2'-deoxyribose). Nucleosides can be further modified with a 5'-phosphate or phosphate-like group, and RNA oligomers contain nucleotides linked via a phosphate or phosphate-like bond from the 5' to the 3' position. Nucleosides can be modified in several ways, including modification to the ribose moiety, the base moiety, or the phosphate moiety, which produces compounds called “nucleoside analogs” (NAs).

[0003] Na has a long and rich history in the field of medicinal chemistry and is also considered a tool compound in chemical biology. Naturally occurring nucleosides are involved in numerous biological processes, making them a unique and valuable starting point for drug design. Synthetic na has been designed to mimic its natural counterparts (2-18). Single na has been used primarily as a treatment for parasitic, bacterial, and fungal infections, and as a potent and effective anticancer agent. In addition to this "small molecule" form, na can be incorporated into oligomeric structures that can regulate gene expression, thus avoiding the complexities associated with protein inhibition. Such oligomeric structures can include small interfering RNA (siRNA), microRNA (miRNA), inhibitory antisense oligonucleotide (ASO), small activating RNA (saRNA), and messenger RNA (mRNA).

[0004] NA is used in cancer treatment (2, 6) and also occupies the largest class of small molecule antiviral agents (3, 4). Its mechanism involves NA acting as a toxic antimetabolite that inhibits nucleic acid synthesis (4). Alternatively, nucleotide analogs produced by phosphorylation reactions in vivo can inhibit enzymes involved in cancer cell proliferation and viral replication (e.g., DNA / RNA polymerase, ribonucleotide reductase, nucleoside phosphorylation enzymes) (2, 4). NA also shows promise as an epigenetic modulator, and both decitabine and azacitidine inhibit DNA methyltransferase and are approved as cancer treatments (4).

[0005] Decades of advancements in organic and medicinal chemistry have led to the creation of numerous useful nucleoside analogs; however, several challenges remain in the synthesis of further nucleoside analogs. Nucleoside analogs are often synthesized from naturally occurring carbohydrates, which limits the substitution patterns and furanose configurations (e.g., 19-29). Addition of nucleic acid bases to activated ribose derivatives often fails or proceeds with low diastereoselectivity for nucleosides modified at the C2' or C4' position, limiting efficient methods for producing C4'-modified nucleosides, including thionucleosides. The synthesis of nucleosides and nucleoside analogs is described by Meanwell et al., Science, 369, 725-730 (2020) and WO 2021 / 191830. Summary

[0006] This invention relates to a method and intermediates for synthesizing nucleoside analogs in which the C4' position is substituted.

[0007] In one embodiment, a halohydrin ketone compound having the following structure: JPEG2026511608000002.jpg3350 (wherein X may be a halogen; NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; and R1 and R2 may each independently be H, alkyl, ester, or protecting group, or both may be cyclic), and the halohydrin ketone compound is reacted with a soft nucleophile to obtain a nucleoside analog in which the C4' position of the following structure is substituted: A method is provided for synthesizing a nucleoside analog with a C4' position substituted by generating a formula in which NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; Nu may be a soft nucleophile; R1 may be H, alkyl, ester, protecting group, or covalently bonded to Nu; and R2 may also be H, alkyl, ester, protecting group, or covalently bonded to Nu).

[0008] In another embodiment, R1 and R2 may each be independently -OH, -OC(CH3)2O-, -(CH2)3-, -CH2SCH2-, or -CH2OCH2-.

[0009] In another embodiment, the halohydrin ketone compound is JPEG2026511608000004.jpg70105 may be any one of the following (wherein X may be a halogen; NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; Y may be CH2, O, S, or NR; wherein R may be an alkyl, aryl, acyl, or carboxylate; and Z may be an alcohol protecting group). In some embodiments, R may be tert-butyloxycarbonyl, benzyloxycarbonyl, or propargyl.

[0010] In another embodiment, the halohydrin ketone compound is It may be any of JPEG2026511608000005.jpg46127 (wherein NB may be optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, and X may be halogen).

[0011] In another embodiment, the halohydrin ketone compound is It may be any of JPEG2026511608000006.jpg41120 (wherein NB may be optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, X may be halogen, Y may be CH2, O, S, NR, wherein R may be alkyl, aryl, acyl, or carboxylate).

[0012] In another embodiment, the halohydrin ketone compound is It may be JPEG2026511608000007.jpg32125.

[0013] In another embodiment, a method for synthesizing a nucleoside analog substituted at the C4' position is a halohydrin ketone compound having the following structure: JPEG2026511608000008.jpg3454 (wherein X may be halogen; NB may be optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; and PG may be a protecting group), and reacting the halohydrin ketone compound with a soft nucleophile to produce a nucleoside analog substituted at the C4' position having the following structure: JPEG2026511608000009.jpg4059 (wherein NB may be optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; Nu may be a soft nucleophile; and PG may be a protecting group that can optionally form a covalent bond with Nu).

[0014] In another embodiment, the protecting group of the alcohol may be an acetonide, a silyl protecting group, an alkyl protecting group, or an aryl protecting group.

[0015] In another embodiment, the protecting group of the alcohol may be tert-butyl(dimethyl)silyl (TBS), triisopropylsilyl (TIPS), trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), triethylsilyl (TES), 2-(trimethylsilyl)ethoxymethyl ether (SEM), tetrahydropyranyl (THP) ether, acetate (Ac), pivalate (Piv), methoxymethyl, or p-methoxybenzyl (PMB).

[0016] In another embodiment, the soft nucleophile may be MeCN, N3 - , CN - , or MeNO2.

[0017] In another embodiment, the halogen may be F.

[0018] In another embodiment, NB is phenyl, biphenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, dihydrobenzopyranyl, 1,4-benzodioxanyl, furan, thiophene, pyrrole, oxazole, thiazole, imidazole, pyrazole, isoxazole, isothiazole, 1,2,3-oxadiazole, triazole, 1,3,4-thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, 2,6-dichloropyrimidinepyrazine, 1,3,5-triazine, imidazole, benzimidazole, benzoxazole, benzothiazo It may also be ethanol, indidine, indole, isoindole, benzofuran, benzothiophene, 1H-indazole, purine, 4H-quinolidine, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, pteridine, uracil, thymine, deazadenine, phthalimide, adenine, 5,6-dihydrouracil, 5-methylcytosine, 5-hydroxymethylcytosine, 5,5,5-trifluoromethylthymine, 5-fluorouracil, 2-thiouracil, 4-methylbenzimidazole, hypoxanthine, 7-deazaguanine, or 7-deazaadenine.

[0019] In another embodiment, the nucleoside analog may be a monomer in LOK nucleic acid (LNA), a small molecule therapeutic agent, or an oligonucleotide therapeutic agent.

[0020] In another embodiment, the nucleoside analog may contain a nitrile moiety and may be reduced to an aldehyde.

[0021] In another embodiment, the nucleoside analog with the C4' position substituted is JPEG2026511608000010.jpg85128 is also acceptable.

[0022] In another embodiment, the following nucleoside analogs are found with the C4' position substituted: The file JPEG2026511608000011.jpg83125 is provided.

[0023] The summary of this invention does not necessarily describe all of its features. Detailed description

[0024] This disclosure provides, in part, a method for synthesizing nucleoside analogs, such as nucleoside analogs (NA) with a substituted C4' position.

[0025] In some embodiments, the disclosure provides a method for synthesizing C4'-substituted NA by reducing a halohydrin ketone compound with a soft nucleophile.

[0026] In some embodiments, the present disclosure relates to a halohydrin ketone compound having the following structure: JPEG2026511608000012.jpg3759 (wherein X may be a halogen; NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; and R1 and R2 may each independently be H, alkyl, ester, or protecting group, or both may be cyclic) are reacted, and the halohydrin ketone compound is reacted with a soft nucleophile to obtain a nucleoside analog in which the C4' position of the following structure is substituted: The present invention provides a method for synthesizing C4'-substituted NA by generating a formula in which NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; Nu may be a soft nucleophile; R1 may be H, alkyl, ester, protecting group, or covalently bonded to Nu; and R2 may also be H, alkyl, ester, protecting group, or covalently bonded to Nu).

[0027] A nucleophile is an atom or molecule that contains an electron pair available to bond with the nucleus of another molecule to form a covalent bond. Examples include neutral molecules with lone pairs of electrons or deprotonated molecules. pKa can relate to both nucleophilic and leaving group capabilities. The pKa values ​​of many common species are listed below, for example (from http: / / ccc.chem.pitt.edu / wipf / MechOMs / evans_pKa_table.pdf). JPEG2026511608000014.jpg187130JPEG2026511608000015.jpg95130JPEG2026511608000016.jpg193130

[0028] A "soft nucleophile" refers to a nucleophile with a low charge density, highly polar orbitals, and the ability to overlap with the nuclear orbitals of other molecules to form covalent bonds. In this specification, a "soft nucleophile" is a nucleophile capable of causing a reversible addition reaction to a ketone or oxocarbenium ion. Therefore, a suitable soft nucleophile must have sufficient nucleophilicity to cause an addition reaction to a ketone or oxocarbenium ion, and must also have a leaving group sufficient to make the addition reaction reversible. In some embodiments, the soft nucleophiles used herein may have a pKa / pKaH of about 0 to about 20, or any value in between, such as 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, or 20. "About" means a variation (plus or minus) from a value or range of 5% or less, such as 0.5%, 1%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, and 5.0%. As can be understood, the pKa of functional groups such as alkynes or carbonyl compounds can be modified by the addition of Lewis acids. Examples of such functional groups include, but are not limited to, water, alcohols, hydrogen sulfide, thiols, fluorides, hypochlorites, cyanides, azides, thiocyanides, bisulfites, phosphates, nitrites, ammonia, amines, boric acid, boronic acid, boric acid, hydrogen peroxide, carboxylic acids, oximes, hydroxamic acids, hydroperoxides, sulfinic acids, imides, sulfonamides, guanidine, hydrazides, hydrazines, amidines, imidazoles, 1,2,3-triazoles, pyridones, indenes, cyclopentadienes, 1,3-dicarbonyls, malononitriles, 3-oxonitriles, sulfoniums, phosphoniums, stabilized phosphonates, and nitroalkanes. As can be understood, suitable soft nucleophiles can cause reversible addition reactions to ketones or oxocarbenium ions even if their pKa is outside the range of 0-20. Examples of soft nucleophiles include MeCN and N3. - 5CN- This includes, but is not limited to, MeNO2, etc. On the other hand, “hard nucleophiles” are highly reactive atoms or molecules such as Grignard reagents and alkyllithiums, and are particularly excluded. In this specification, “Nu” refers to a soft nucleophile that can attack the halohydrin ketone compound before cyclization or can bind to the oxonium ion produced by the cyclization of the halohydrin ketone compound.

[0029] A "halohydrin ketone compound" refers to a compound containing a ketone group and a functional group in which a halogen and a hydroxyl group are bonded to adjacent groups. Therefore, in some embodiments, the halohydrin ketone compound has the following general structure: The formula may include JPEG2026511608000017.jpg3960. In the formula, X may be a halogen, NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, and R1 and R2 may each independently be H, alkyl, ester, or protecting group.

[0030] In some embodiments, R1 and R2 may each be independently OH, -OC(CH3)2O-, -(CH2)3-, -CH2SCH2-, or -CH2OCH2-.

[0031] In some embodiments, the halohydrin ketone compound has the following general structure for use as an intermediate in the synthesis of nucleosides or their analogues: JPEG2026511608000018.jpg79113 may have the following characteristics: X may be a halogen, NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, Y may be CH2, O, S, or NR, R may be an alkyl, aryl, acyl, or carboxylate (e.g., tert-butyloxycarbonyl, benzyloxycarbonyl, or propargyl), and Z may be an alcohol protecting group including, but not limited to, an acetonide, a sylyl protecting group, an alkyl protecting group, or an aryl protecting group (including cyclic or acyclic).

[0032] In some embodiments, the halohydrin ketone compound has the following general structure: The formula may include JPEG2026511608000019.jpg47136, where NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, and X may be a halogen.

[0033] In some embodiments, the halohydrin ketone compound has the following general structure: JPEG2026511608000020.jpg41122 may have the formula, where NB is optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, X is a halogen, Y may be CH2, O, S, or NR, where R may be alkyl, aryl, acyl, or carboxylate (for example, R may be tert-butyloxycarbonyl, benzyloxycarbonyl, or propargyl).

[0034] A "nucleoside" refers to a glycosylamine having a nitrogenous base ("nucleic acid base" or "NB") or a sugar ring (e.g., ribose or deoxyribose), where the anomeric carbon is bonded via a glycosidic bond to the N9 of a purine (e.g., adenine or guanine) or the N1 of a pyrimidine (e.g., cytosine, thymine, or uracil). Nucleosides include both L-nucleoside isomers and D-nucleoside isomers. Examples of nucleosides include cytidine, uridine, adenosine, guanosine, thymidine, and inosine.

[0035] Nucleoside analogs (NAs) are compounds that are structurally similar to naturally occurring nucleosides. NAs may include, but are not limited to, compounds in which the C1', C2', C3', C4', and / or C5' positions of the sugar ring are modified, based on conventional numbering as shown below. JPEG2026511608000021.jpg4961

[0036] In some embodiments, NA may exist as a free triol or may be phosphorylated at the C3' and / or C5' positions. In some embodiments, NA may include, but is not limited to, compounds having saturated or unsaturated carbocyclic rings. In some embodiments, NA may contain nitrogen in the sugar ring and / or an NR group, for example as a substitution for naturally occurring oxygen, and / or R may be alkyl, aryl, alkynyl, or benzyl, but is not limited to these. In some embodiments, the nucleoside analogs disclosed herein may be modified to function as phosphoramide or phosphonamide compounds, e.g., "ProTide". This comprises a 5'-nucleoside monophosphate with two hydroxyl groups masked by an amino acid ester, and an aryloxy component that is enzymatically metabolized to produce free 5'-monophosphate, which, upon delivery to cells, is further converted to the active 5'-triphosphate form of the nucleoside analog. In some embodiments, NA containing sulfur in the sugar ring is specifically excluded, for example as a substitution for naturally occurring oxygen.

[0037] The base of "NB" or the nucleic acid base or NA may be an aryl, arylalkyl, heteroaryl, or heteroarylalkyl bonded to a carbon or nitrogen atom from the C1 position. NB may also be modified, for example, one or more of the following: 5,6-dihydrouracil, 5-methylcytosine, 5-hydroxymethylcytosine, 5,5,5-trifluoromethylthymine, 5-fluorouracil, 2-thiouracil, 4-methylbenzimidazole, hypoxanthine, 7-deazaguanine, 7-deazaadenine, indole, imidazole, triazole, pyrrole, and pyrazole. As understood, the enantiomer of the aldol product (halohydrin) can be produced using a D-proline catalyst, which may also be used to prepare the enantiomer NA.

[0038] "Alkyl" refers to a linear or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, generally unsaturated, and containing, for example, 1 to 10 carbon atoms, or any value in between, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, which are bonded to the rest of the molecule by single bonds. In some embodiments, alkyl may refer to a linear or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, generally unsaturated, and containing 1 to 6 carbon atoms, or any value in between, e.g., 1, 2, 3, 4, 5, or 6 carbon atoms, which are bonded to the rest of the molecule by single bonds. Unless otherwise specified herein, alkyl groups may be optionally substituted with one or more substituents described herein. Unless otherwise specified herein, this substitution may occur on any carbon of the alkyl group, as understood.

[0039] "Aryl" means a monocyclic or bicyclic aromatic ring containing only carbon atoms, including, for example, 5- to 14-membered rings, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14-membered rings. Examples of aryl groups include phenyl, biphenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, dihydrobenzopyranyl, and 1,4-benzodioxanyl. Unless otherwise specified herein, the term "aryl" means an aryl group optionally substituted with one or more substituents as described herein.

[0040] A "heteroaryl" refers to a single or condensed aromatic ring group containing one or more heteroatoms within the ring, such as N, O, S, and 5 to 14 members, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 members. Examples of heteroaryl groups include furan, thiophene, pyrrole, oxazole, thiazole, imidazole, pyrazole, isoxazole, isothiazole, 1,2,3-oxadiazole, triazole (e.g., 1,2,3-triazole or 1,2,4-triazole), 1,3,4-thiadiazole, tetrazole, pyrazole, pyridine, pyridazine, pyrimidine, 2,6-dichloropyrimidine, pyrazine, 1,3,5-triazine, imidazole, benzimidazole, benzoxazole, benzothiazole, indidine, indole, isoindole, benzofuran, benzothiophene, 1H-indazole, purine, 4H-quinolidine, quinoline, isoquinoline, cinolin, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, pteridine, uracil, thymine, deazadenine, phthalimide, and adenine. Unless otherwise specified herein, the term “heteroaryl” means a heteroaryl group optionally substituted with one or more substituents as described herein.

[0041] "Arylalkyl" refers to the compound of formula -R a R b It means the base of, and in the formula, R a C as described in this specification 1-10is an alkyl group, and R b is one or more aryl groups as described herein. The arylalkyl group may be optionally substituted as described herein. Examples of arylalkyl groups include, but are not limited to, benzyl group, phenethyl group, phenylpropyl group, (4-methylphenyl)methyl group, (4-methylphenyl)ethyl group, (2-methylphenyl)methyl group, (2,4,6-trimethylphenyl), (4-fluorophenyl)methyl group, and (4-trifluoromethylphenyl)methyl group, etc.

[0042] "Heteroarylalkyl" refers to a group of the formula -R a R c wherein R a is a C 1-10 alkyl group as described herein, and R c is one or more heteroaryl moieties as described herein. The heteroarylalkyl group may be optionally substituted as described herein. Examples of heteroarylalkyl groups include, but are not limited to, furanylmethyl, thiophenylmethyl, pyridylmethyl, imidazolylmethyl, and uridinylmethyl, etc.

[0043] "Acyl" refers to a group of the formula -C(O)R a wherein R a is a C 1-10 alkyl group or C 1-6 alkyl group as described herein. The alkyl group may be optionally substituted as described herein.

[0044] "Ester" means a carbon atom (A-COOA') double-bonded to a first oxygen atom and single-bonded to a second oxygen atom, wherein A can be any carbon-bonded group. Suitable ester groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and pentyl, etc.

[0045] "Cylindrical" refers to a group in which one or more atomic sequences are bonded together to form a ring. The atoms in a cyclic group may be carbon, oxygen, nitrogen, silicon, or sulfur. The size of the ring varies, for example, containing three or eight atoms, or any number of atoms in between, such as three, four, five, six, seven, or eight. Suitable cyclic groups include, but are not limited to, 1,3-dioxolane, 1,3-dioxanone, oxazoline, and tetrahydrofuran.

[0046] Halogens include bromine, chlorine, fluorine, and iodine, and are represented by "X" in the chemical structures disclosed herein. In some embodiments, halogens may include chlorine or fluorine. In some embodiments, the halogen may be fluorine. In some embodiments, the halogen may be chlorine. Therefore, "halo" refers to bromine, chlorine, fluorine, and iodine, etc. Halides are halogen atoms that have a negative charge.

[0047] As used herein, “protecting group” means a reversibly formed derivative of a functional group present in a molecule, which temporarily binds to and reduces the reactivity of the protected functional group so that the molecule does not react under synthetic conditions subjected to one or more subsequent steps. Protecting groups are known in the art and are described herein. As will be understood, those skilled in the art can readily determine a protecting group suitable for a particular synthesis.

[0048] In some embodiments, "suitable alcohol protecting groups" or "alcohol protecting groups" include, but are not limited to, acetonides, silyl protecting groups, alkyl protecting groups, or aryl protecting groups (including cyclic and acyclic), such as silyl ethers, tert-butyl(dimethyl)silyl (TBS), triisopropylsilyl (TIPS), trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), triethylsilyl (TES), 2-(trimethylsilyl)ethoxymethyl ether (SEM), etc.; acetate (Ac), pivalate (Piv) or other ester or carbonate protecting groups, etc.; benzyl, aryl, methoxymethyl or p-methoxybenzyl (PMB) or other ether protecting groups, etc.; tetrahydropyranyl (THP) ether or other related protecting groups, etc.

[0049] "Covalent bonding to Nu" means the conjugation of an "R" group as described herein, such as R, R1, R2, R3, or R4, with a soft nucleophile as described herein. Examples of such conjugation include, but are not limited to, attacking a C3' or C5' hydroxyl group with a soft nucleophile to form a covalent bond, such as attacking a nitrilium group to form an oxazoline.

[0050] "Optional" or "optionally" means that the events or situations described below may or may not occur, and that the description includes both cases where the event or situation occurs once or more and cases where it does not occur. For example, "optionally substituted alkyl group" means that the alkyl group may or may not be substituted, that the description includes both substituted alkyl groups and unsubstituted alkyl groups, and that the alkyl group may be substituted once or more. Examples of optional substituted alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, and tert-butyl. Examples of suitable optional substituents include H, F, Cl, CH3, OH, OCH3, CF3, CHF2, CH2F, CN, halo, and C 1-10Examples include, but are not limited to, alkoxy groups. Similarly, "optionally substituted aryl or heteroaryl groups" means that the aryl or heteroaryl group may be substituted or unsubstituted, that the description includes both substituted and unsubstituted aryl or heteroaryl groups, and that the aryl or heteroaryl group may be substituted one or more times. Examples of suitable optional substituents include H, F, Cl, CH3, OH, OCH3, CF3, CHF2, CH2F, CN, halo, and C 1-10 Examples include, but are not limited to, alkoxy groups.

[0051] In some embodiments, the methods disclosed herein allow for the rapid acquisition of intermediates in the synthesis of nucleosides or their analogues, for example, in amounts greater than about 10 g to about 400 g, or any value in between, such as 10 g, 15 g, 20 g, 25 g, 50 g, 75 g, 100 g, 125 g, 150 g, 200 g, 250 g, 300 g, 350 g, or 400 g, with good enantioselectivity and / or yield. Accordingly, the methods disclosed herein can be used for process-scale production of NA with modified C4' position.

[0052] In some embodiments, this disclosure includes a method for synthesizing a C4'-substituted nucleoside analog according to, for example, Scheme 1: In formula JPEG2026511608000022.jpg37116, X may be a halogen; NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; R1 and R2 may each independently be H, alkyl, ester, or protecting group, or together form a cyclic group; the nucleophile and Nu may be soft nucleophiles as described herein; R3 may be covalently bonded to H, alkyl, ester, protecting group, or Nu; and R4 may also be covalently bonded to H, alkyl, ester, protecting group, or Nu.

[0053] In some embodiments, this disclosure includes a method for synthesizing a C4'-substituted nucleoside analog according to, for example, Scheme 2: In formula JPEG2026511608000023.jpg34127, X may be a halogen; NB may be an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl; Nu may be a soft nucleophile as described herein; and PG may be a protecting group that can form a covalent bond with Nu in the product. In some embodiments, the protecting group may be acetonide (CH3)2C, siloxane-Si(CH3)2, O-Si(CH3)2-, benzyl group, p-methoxybenzyl group, TBS, or other silyl protecting group.

[0054] In some embodiments, the methods disclosed herein allow direct access to sequences of C4'-modified NAs, including but not limited to LOK nucleic acids (LNAs).

[0055] In some embodiments, an aldehyde product can be obtained by reducing a nucleoside analog in which the C4' position containing a nitrile moiety is substituted.

[0056] In some embodiments, C4'-substituted nucleoside analogs according to this disclosure include: JPEG2026511608000024.jpg86131

[0057] In some embodiments, the methods disclosed herein may be useful for drug design.

[0058] In some embodiments, the nucleoside analogs disclosed herein may be used as small molecule therapeutic agents or as monomers in oligonucleotide therapeutic agents.

[0059] In some embodiments, the methods disclosed herein may be useful for constructing diversity libraries. For example, the methods described herein can be used to construct a larger collection of nucleic acids (NAs) with altered C4' positions (e.g., a focused screening library).

[0060] As used herein, the singular forms “a,” “and,” and “the” include multiple references unless the context explicitly states otherwise. For example, “one compound” refers to one or more such compounds. Throughout this application, the terms “compound” or “compounds (plural)” refer to the compounds discussed herein, and also include their precursors and derivatives. The compounds of the present invention may contain one or more chiral centers and thus may exist as racemates and racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers. Additional chiral centers may exist due to the properties of various substituents on the molecule. Each of these chiral centers independently produces two optical isomers. Unless otherwise specified, all potential optical isomers and diastereomers that may exist in mixtures and as pure or partially purified compounds are included in the scope of the present invention. The formulas, structures, or names of compounds described herein without specifying a particular configuration mean that they encompass all existing isomers and mixtures of any proportion thereof, as stated above. Where a stereochemistry is specified, the present invention means that the particular isomer is included in its pure form or as part of a mixture of other isomers in any proportion. A single enantiomer, i.e., an optically active compound, can be obtained by asymmetric synthesis or racemic resolution. Racemic resolution can be achieved by conventional methods, for example, crystallization in the presence of a resolving agent, chromatography using a chiral HPLC column, or by derivatizing the racemic mixture with a resolving reagent to produce a diastereomer, separating the diastereomer by chromatography, and then removing the resolving agent to obtain the original compound in an enantiomer-enriched form. These steps can be repeated as needed to increase the enantiomer purity of the compound. Where a compound described herein contains an olefinic double bond or other geometrically asymmetric center, unless otherwise specified, the compound shall include cis, trans, Z-, and E- configurations. Similarly, all tautomers shall also be included.

[0061] The raw materials can be obtained from commercially available suppliers, prepared from commercially available organic compounds, and / or prepared using known synthetic methods.

[0062] The present invention will be further explained by the following examples. [Examples]

[0063] Materials and methods

[0064] Nuclear magnetic resonance (NMR) spectra were recorded using deuterated chloroform (CDCl3), deuterated methanol (CD3OD), deuterated acetone ((CD3)2CO), deuterated acetonitrile (CD3CN), or deuterated dimethyl sulfoxide (DMSO-d6) as solvents. The signal position (δ) is shown in ppm from tetramethylsilane (δ 0) and measured relative to the signal of the solvent. 1 H NMR:CDCl3:δ 7.26; CD3OD:δ 3.31; (CD3)2CO:δ 2.05; CD3CN: δ 1.96; DMSO-d6:δ 2.50; 13 ¹³C NMR: CDCl3: δ 77.16; CD3OD: δ 49.00; (CD3)2CO: δ 29.84; CD3CN: δ 1.32; DMSO-d6: 39.5). The coupling constants (J values) are shown in Hertz (Hz) and displayed to the nearest 0.1 Hz. 1 The 1H NMR spectral data are summarized in a table in the following order: multiplicity (s: singlet, d: doublet, t: triplet, q: quartet, sept: septet, m: multiplet, br: broad), coupling constant, and proton number. NMR spectra were recorded using a Bruker Avance 600, Bruker 400 (400 MHz), or Bruker 500 (500 MHz) equipped with a QNP or TCI cryoprobe (600 MHz). The diastereomer ratio (dr) is crude. 1 Based on 1H NMR analysis. 1 The attribution of H is, 1 H- 1 This analysis was based on H-COSY and nOe spectra. 13The assignment of C is based on the analysis of the HSQC spectrum.

[0065] Examples JPEG2026511608000025.jpg26141

[0066] InCl3 (35.0 mg, 158 μmol, 1 equivalent) was added to a MeCN solution (1.58 mL, 0.1 mol reaction concentration) of the stirred starting ketone (50.0 mg, 158 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 18 hours, after which a saturated aqueous solution of sodium bicarbonate (2 mL) was added to stop the reaction. The mixture was extracted with ethyl acetate (10 x 5 mL), the combined organic extract was dried (MgSO4), and concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain a pure product (25.0 mg, 84.1 μmol, 53%).

[0067] 1 H NMR (500 MHz, MeCN-d6) δ 9.13 (1H, br s), 7.35 (1H, q, J = 1.0 Hz), 5.65 (1H, d, J = 8.2 Hz), 4.71 (1H, d, J = 5.9 Hz), 4.31 (1H, m), 3.80 (1H, dd, J = 11.8, 5.2 Hz), 3.71 (1H, d, J = 7.8 Hz), 3.60 (1H, dd, J = 11.8, 6.3 Hz), 3.36 (1H, app t, J = 6.1 Hz), 2.01 (3H, s), 1.87 (3H, d, J = 1.0 Hz). JPEG2026511608000026.jpg46150

[0068] InCl3 (3.5 mg, 15.8 μmol, 1 equivalent) was added to a 0.32 mL allyl nitrile solution (0.05 mol reaction concentration) of the stirred starting ketone (5.00 mg, 15.8 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 18 hours, after which a saturated aqueous solution of sodium bicarbonate (1 mL) was added to stop the reaction. The mixture was extracted with ethyl acetate (5 x 2 mL), the combined organic extract was dried (MgSO4), and concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain the pure product.

[0069] 1 H NMR (500 MHz, MeCN-d6) δ 9.05 (1H, br s), 7.35 (1H, s), 6.89 (1H, dq, J = 15.8, 6.9 Hz), 6.08 (1H, dq, J = 15.8, 1.6 Hz), 5.57 (1H, d, J = 8.4 Hz), 4.75 (1H, d, J = 5.8), 4,32 (1H, m), 3.83 (1H, dd, J = 11.9, 3.0 Hz), 3.75 (1H, br d, J = 6.3 Hz), 3.63 (1H, m), 3.36 (1H, br t, J = 5.1 Hz), 1.94 (3H, dd, J = 6.9, 1.6 Hz), 1.88 (3H, d, J = 0.9 Hz). JPEG2026511608000027.jpg48151

[0070] To a benzonitrile solution (0.32 mL, 0.05 mol reaction concentration) of the stirred starting ketone (5.00 mg, 15.8 μmol, 1 equivalent), InCl3 (3.5 mg, 15.8 μmol, 1 equivalent) was added. After stirring the reaction mixture at room temperature for 18 hours, a saturated aqueous solution of sodium bicarbonate (1 mL) was added to stop the reaction. The mixture was extracted with ethyl acetate (5 x 2 mL), the combined organic extract was dried (MgSO4), and concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain the pure product.

[0071] 1 H NMR (500 MHz, MeCN-d3) δ 8.06 (2H, d, J = 7.6 Hz), 7.65 (1H, t, J = 7.6 Hz), 7.55 (2H, t, J = 7.6 Hz), 7.38 (1H, s), 5.66 (1H, d, J = 8.4 Hz), 4.96 (1H, d, J = 5.7 Hz), 4.29 (1H, m), 3.93 (1H, m), 3.73 (1H, br d, J = 11.7 Hz), 1.87 (3H, s). JPEG2026511608000028.jpg26138

[0072] NaCN (46.49 mg, 949 μmol, 3 equivalents) was added to a methanol solution (6.3 mL, reaction concentration of 0.05 mol) of the stirred starting ketone (100.0 mg, 316 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 18 hours and then concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain a pure product (70.0 mg, 0.22 mmol, 68%).

[0073] 1 H NMR (500 MHz, MeOH-d4) δ 7.32 (1H, s), 5.90 (1H, br s), 4.61 (1H, br d, J = 5.7 Hz), 4.38 (1H, d, J = 5.7 Hz), 4.27 (2H, abq, J = 10.1 Hz), 1.90 (3H, s), 1.64 (3H, s), 1.58 (3H, s). JPEG2026511608000029.jpg26140

[0074] The stirring starting ketone (1.00 g mg, 3.16 mmol, 1 equivalent) was dissolved in an aqueous solution (63.2 mL, reaction concentration of 0.05 mol) and NaCN (170 mg, 3.48 mmol, 1.1 equivalents) was added. The reaction mixture was stirred at room temperature for 2 hours, and then ferrous sulfate (483 mg, 1.74 mmol, 0.55 equivalents) was added to stop the reaction. The mixture was filtered, and the filtrate was extracted with ethyl acetate (10 x 50 mL). The combined organic extracts were dried (magnesium sulfate) and concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain a pure product (492 mg, 1.52 mmol, 48%).

[0075] 1 H NMR (500 MHz, MeOH-d4) δ 7.32 (1H, s), 5.90 (1H, br s), 4.61 (1H, br d, J = 5.7 Hz), 4.38 (1H, d, J = 5.7 Hz), 4.27 (2H, abq, J = 10.1 Hz), 1.90 (3H, s), 1.64 (3H, s), 1.58 (3H, s). JPEG2026511608000030.jpg27123

[0076] NaCN (2.8 mg, 58 μmol, 3 equivalents) was added to an aqueous solution (0.19 mL, 0.1 molar reaction concentration) of the stirred starting ketone (5.0 mg, 19 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 2 hours and then concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain a pure product (4.6 mg, 17 μmol, 89%).

[0077] 1H NMR (500 MHz, MeCN-d3) δ 7.75 (1H, d, J = 2.4 Hz), 7.60 (1H, d, J = 1.6 Hz), 7.32 (1H, dd, J = 2.4, 1.6 Hz), 6.02 (1H, br s), 4.70 (1H, dd, J = 4.9, 3.6 Hz), 4.44 (1H, d, J = 4.9 Hz), 4.28 (1H, d, J = 10.2 Hz), 4.06 (1H, d, J = 3.6 Hz), 4.03 (1H, d, J = 1.02 Hz), 1.57 (3H, s), 1.54 (3H, s). JPEG2026511608000031.jpg27122

[0078] NaCN (2.8 mg, 58 μmol, 3 equivalents) was added to a water / methanol solution (1:1, 0.19 mL, reaction concentration of 0.1 molars) of the stirred starting ketone (5.0 mg, 19 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 4 hours and then concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain a pure product (4.6 mg, 17 μmol, 89%).

[0079] 1 H NMR (500 MHz, MeCN-d3) δ 7.75 (1H, d, J = 2.4 Hz), 7.60 (1H, d, J = 1.6 Hz), 7.32 (1H, dd, J = 2.4, 1.6 Hz), 6.02 (1H, br s), 4.70 (1H, dd, J = 4.9, 3.6 Hz), 4.44 (1H, d, J = 4.9 Hz), 4.28 (1H, d, J = 10.2 Hz), 4.06 (1H, d, J = 3.6 Hz), 4.03 (1H, d, J = 1.02 Hz), 1.57 (3H, s), 1.54 (3H, s). JPEG2026511608000032.jpg26140

[0080] NaCN (2.4 mg, 50 μmol, 3 equivalents) was added to an aqueous solution (0.17 mL, 0.1 molar reaction concentration) of the stirred starting ketone (5.0 mg, 17 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 2 hours and then concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain a pure product (4.5 mg, 15 μmol, 88%).

[0081] 1 H NMR (500 MHz, MeCN-d3) δ 9.10 (1H, br s), 7.36 (1H, d, J = 8.2 Hz), 5.83 (1H, br s), 5.66 (1H, d, J = 8.2 Hz), 4.54 (1H, br d, J = 5.5 Hz), 4.31 (1H, d, J = 10.2 Hz), 4.12 (1H, d, J = 10.2 Hz), 4.06 (1H, d, J = 5.5 Hz), 4.00 (1H, br s) 1.55 (3H, s), 1.53 (3H, s). JPEG2026511608000033.jpg30121

[0082] Diisobutylaluminum hydride (1.63 mL, 0.91 mol / 1.48 mmol / 4 equivalents in dichloromethane) was added dropwise to a stirred solution of nitrile (120 mg, 0.371 mmol) in dichloromethane (3.71 mL) at -78°C. The reaction mixture was stirred at -78°C for 1 hour, after which a saturated aqueous solution of potassium sodium tartrate (3 mL) was added to quench the reaction. The mixture was stirred at room temperature for 1 hour and then extracted with ethyl acetate (10 mL x 3). The crude product was purified by silica chromatography and eluted with hexane / (ethyl acetate solution of 1% isopropyl alcohol) (1:1 -> 1:2). The product, along with the recovered starting material (108 mg, 0.334 mmol / 90%), was obtained as a colorless oil (12.1 mg, 3.71 mmol / 10%).

[0083] 11H NMR (600 MHz, D3CCN): TIFF2026511608000034.tif429.11 (s, 1H, NH), 7.38 (d, J = 1.4 Hz, 1H), 5.74 (s, 1H), 5.61 (s, 1H), 4.48 (s, 1H), 4.42 (d, J = 11.0 Hz, 1H), 4.01 (d, J = 11.0 Hz, 1H), 3.93 (s, 1H), 1.88 (d, J = 1.3 Hz, 3H), 1.51 (s, 3H), 1.43 (s, 3H); 13 ¹³C NMR (126 MHz, D3CCN): HRMS (ESI + ) calcd for [C 14 H 18 N2O7+H] + 327.1187 m / z found: 327.1187 m / z ( TIFF2026511608000036.tif420.8 ppm). JPEG2026511608000037.jpg25136

[0084] TMS azide (9.1 mg, 79 μmol, 5 equivalents) was added to a CH2Cl2 solution (0.16 mL, 0.1 mol reaction concentration) of the stirred starting ketone (5.0 mg, 16 μmol, 1 equivalent). The reaction mixture was stirred at room temperature for 18 hours and then concentrated under vacuum. The crude product was purified by silica chromatography (ethyl acetate / hexane) to obtain the pure product (3.75 mg, 12 μmol, 75%).

[0085] 1H NMR (500 MHz, MeCN-d3) δ 7.35 (1H, q, J = 1.0 Hz), 6.22 (1H, d, J = 8.0 Hz), 4.67 (1H, dd, J = 8.0, 4.1 Hz), 4.14 (1H, d, J = 4.1 Hz), 4.11 (1H, d, J = 12.8 Hz), 4.01 (1H, d, J = 12.8 Hz), 1.91 (3H, d, J = 1.0 Hz), 1.50 (3H, s), 1.47 (3H, s). JPEG2026511608000038.jpg25142

[0086] Under an argon atmosphere, tert-butoxy potassium (1.06 g, 9.48 mmol, 3 equivalents) and cesium carbonate (2.06 g, 6.32 mmol, 2 equivalents) were added to anhydrous nitromethane (32 mL) and heated at 65°C for 10 minutes. A suspension was then formed, and ketone (1.00 g, 3.16 mmol, 1 equivalent) was added to the reaction mixture all at once. The mixture was stirred at 65°C for a further 2.5 hours. After cooling, the reaction was stopped with saturated NH4Cl, diluted with ethyl acetate, and washed with water and saturated brine. The organic phase was dried over NaSO4, filtered, and the solvent was removed under reduced pressure. The pure product (0.621 g, 1.74 mmol, 55%) was obtained by flash chromatography (ethyl acetate / hexane) of the residue.

[0087] 1 H NMR (CD3CN, 500 MHz) δ 7.19 (1H, s), 5.92 (1H, d, J = 1.5 Hz), 5.31 (1H, dd, J = 12.4, 1.6 Hz), 5.01 (1H, d, J = 12.4 Hz), 4.44 (1H, t, J = 4.6 Hz), 4.35 - 4.22 (2H, m), 4.06 - 3.98 (2H, m), 2.26 - 2.20 (1H, m), 1.83 (3H, d, J = 1.4 Hz), 1.56 (3H, s), 1.46 (3H, s).

[0088] References 1. G. M. Blackburn, Gait, M. J., Loakes, D., Williams, D. M., Ed., Nucleic Acids in Chemistry and Biology, (Royal Society of Chemistry, Cambridge, UK, 2006), pp. 503. 2. C. M. Galmarini, J. R. Mackey, C. Dumontet. Nucleoside Analogues and Nucleobases in Cancer Treatment. Lancet Oncol. 3, 415 - 424 (2002). 3. E. De Clercq. Highlights in Antiviral Drug Research: Antivirals at the Horizon. Med. Res. Rev. 33, 1215 - 1248 (2013). 4. L. P. Jordheim, D. Durantel, F. Zoulim, C. Dumontet. Advances in the Development of Nucleoside and Nucleotide Analogues for Cancer and Viral Diseases. Nat. Rev. Drug Discov. 12, 447 - 464 (2013). 5. D. M. Huryn, M. Okabe. AIDS - Driven Nucleoside Chemistry. Chem. Rev. 92, 1745 - 1768 (1992). 6. J. Shelton et al. Metabolism, Biochemical Actions, and Chemical Synthesis of Anticancer Nucleosides, Nucleotides, and Base Analogs. Chem. Rev. 116, 14379 - 14455 (2016). 7. B. Ewald, D. Sampath, W. Plunkett. Nucleoside Analogs: Molecular Mechanisms Signaling Cell Death. Oncogene 27, 6522-6537 (2008). 8. K. L. Seley-Radtke, M. K. Yates. The Evolution of Nucleoside Analogue Antivirals: A Review for Chemists and Non-Chemists. Part 1: Early Structural Modifications to the Nucleoside Scaffold. Antiviral Res. 154, 66-86 (2018). 9. M. K. Yates, K. L. Seley-Radtke. The Evolution of Antiviral Nucleoside Analogues: A Review for Chemists and Non-Chemists. Part II: Complex Modifications to the Nucleoside Scaffold. Antiviral Res. 162, 5-21 (2019). 10.H. Ma et al. Characterization of the Metabolic Activation of Hepatitis C Virus Nucleoside Inhibitor Beta-D-2'-Deoxy-2'-Fluoro-2'-C-Methylcytidine (PSI-6130) and Identification of a Novel Active 5'-Triphosphate Species. J. Biol. Chem. 282, 29812-29820 (2007). 11.E. P. Gillis, K. J. Eastman, M. D. Hill, D. J. Donnelly, N. A. Meanwell. Applications of Fluorine in Medicinal Chemistry. J. Med. Chem. 58, 8315-8359 (2015). 12.J. Deval, M. H. Powdrill, C. M. D'Abramo, L. Cellai, M. Gotte. Pyrophosphorolytic Excision of Nonobligate Chain Terminators by Hepatitis C Virus NS5B Polymerase. Antimicrob. Agents Chemother. 51, 2920-2928 (2007). 13.H. Ohrui. 2'-Deoxy-4'-C-Ethynyl-2-Fluoroadenosine, a Nucleoside Reverse Transcriptase Inhibitor, is Highly Potent Against All Human Immunodeficiency Viruses Type 1 and Has Low Toxicity. Chem. Rec. 6, 133-143 (2006). 14.J. T. Witkowski, R. K. Robins, R. W. Sidwell, L. N. Simon. Design, Synthesis, and Broad Spectrum Antiviral Activity of 1-Beta-D-Ribofuranosyl-1,2,4-Triazole-3-Carboxamide and Related Nucleosides. J. Med. Chem. 15, 1150-1154 (1972). 15.J. Zeidler, D. Baraniak, T. Ostrowski. Bioactive Nucleoside Analogues Possessing Selected Five-Membered Azaheterocyclic Bases. Eur. J. Med. Chem. 97, 409-418 (2015). 16.G. Ni et al. Review of α-Nucleosides: From Discovery, Synthesis to Properties and Potential Applications. RSC Advances 9, 14302-14320 (2019). 17.G. Gumina, G. Y. Song, C. K. Chu. L-Nucleosides as Chemotherapeutic Agents. FEMS Microbiol. Lett. 202, 9-15 (2001). 18.H. Cui et al. Synthesis and Evaluation of alpha-Thymidine Analogues as Novel Antimalarials. J. Med. Chem. 55, 10948-10957 (2012). 19.Chemical Synthesis of Nucleoside Analogues. P. Merino, Ed., (John Wiley & Sons, Inc., 2013), pp. 895. 20.M. Brodszki et al. Synthesis of the Hepatitis B Nucleoside Analogue Lagociclovir Valactate. Org. Process. Res. Dev. 15, 1027-1032 (2011). 21.M. McLaughlin et al. Enantioselective Synthesis of 4′-Ethynyl-2-fluoro-2′-deoxyadenosine (EFdA) via Enzymatic Desymmetrization. Org. Lett. 19, 926-929 (2017). 22.W. T. Markiewicz, M. Wiewiorowski. A New Type of Silyl Protecting Groups in Nucleoside Chemistry. Nucleic Acids Res. 5, s185-s190 (1978). 23.K. R. Campos et al. The Importance of Synthetic Chemistry in the Pharmaceutical Industry. Science 363, eaat0805 (2019). 24.M. Peifer, R. Berger, V. W. Shurtleff, J. C. Conrad, D. W. MacMillan. A General and Enantioselective Approach to Pentoses: a Rapid Synthesis of PSI-6130, the Nucleoside Core of Sofosbuvir. J. Am. Chem. Soc. 136, 5900-5903 (2014). 25.D. Chapdelaine et al. A stereoselective approach to nucleosides and 4'-thioanalogues from acyclic precursors. J. Am. Chem. Soc. 131, 17242-17245 (2009). 26. R. Britton, B. Kang. alpha-Haloaldehydes: Versatile Building Blocks for Natural Product Synthesis. Nat. Prod. Rep. 30, 227-236 (2013). 27. W. Ren et al. Revealing the mechanism for covalent inhibition of glycoside hydrolases by carbasugars at an atomic level. Nat. Commun. 9, 3243 (2018). 28. A. Quintard, J. Rodriguez. Bicatalyzed Three-Component Stereoselective Decarboxylative Fluoro-Aldolization for the Construction of Elongated Fluorohydrins. ACS Catalysis 7, 5513-5517 (2017). 29.TC Britton, ME LeTourneau. (1995). Process for Anomerizing Nucleosides. US 5,420,266. Eli Lilly and Company. 30.Meanwell et al, Science, 2020, 369, 725-730

[0089] All cited references are incorporated herein by reference.

[0090] The present invention has been described in relation to one or more embodiments. However, it will be apparent to those skilled in the art that many modifications and alterations are possible without departing from the scope of the invention as defined in the claims. Accordingly, although various embodiments of the invention are disclosed herein, many adaptations and alterations are possible within the scope of the invention according to the common general knowledge of those skilled in the art. Such alterations include replacing known equivalents for any aspect of the invention with those known in order to achieve the same results in substantially the same way. Numerical ranges include numerical values ​​that define the range. In this specification, the word “comprising” is used as an open-ended term and is substantially synonymous with the phrase “including, but not limited to,” and the word “comprises” has a corresponding meaning. However, it should be understood that wherever the words “contains” or “includes,” or variations of the same etymology are used herein, variations or modifications to the words “constituted” or “composed of” (excluding elements, processes, or components not specified), or variations or modifications to the words “essentially composed of” or “essentially composed of” (limiting to specific materials or described processes, along with those that do not substantially affect the basic and novel features of the claimed invention), are also assumed. “Approximately” means a variation (plus or minus) from a value or range of 5% or less, for example, 0.5%, 1%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, and 5.0%. While the elements of the invention described may be shown in specific embodiments, it should be understood that they can be combined in any way and in any number to create additional embodiments. The various examples and preferred embodiments described should not be construed as limiting the invention to only the expressly described embodiments. This specification should be understood to support and encompass embodiments that combine the expressly described embodiments with any number of disclosed elements and / or preferred elements.Furthermore, any permutations and combinations of all elements described herein should be deemed to be disclosed in the specification unless otherwise stated in the context. References made herein should not be construed as an admission that such references are prior art of the present invention. All publications are incorporated herein by reference in such a manner that it is specifically and individually indicated that each publication is incorporated herein by reference and as if it were fully described herein. The present invention includes substantially the embodiments and variations described herein, and all embodiments referring to the embodiments described herein.

Claims

1. A method for synthesizing a nucleoside analog in which the C4' position is substituted, i) Halohydrin ketone compounds with the following structure: (In the formula, X is a halogen, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, and also, R 1 and R 2 To provide a group in which each is independently H, alkyl, ester, or protecting group, or together forms a cyclic group, and ii) The halohydrin ketone compound is reacted with a soft nucleophile to obtain a nucleoside analog in which the C4' position of the following structure is substituted: (In the formula, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, Nu is a soft nucleophile, R 1 is covalently bonded to H, alkyl, ester, protecting group, or Nu, and also, R 2 The formation of a covalent bond with H, alkyl, ester, protecting group, or Nu. Methods that include...

2. R 1 and R 2 are independently —OH, —OC(CH 3 ) 2 O—, —(CH 2 ) 3 —, —CH 2 SCH 2 —, or —CH 2 OCH 2 —, the method of claim 1

3. The aforementioned halohydrin ketone compound, It is one of the following: During the ceremony, X is a halogen, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, If Y exists, CH 2 The formula is O, S, NR, where R is alkyl, aryl, acyl, or carboxylate, and also, If Z is present, it is an alcohol protecting group. The method according to claim 1.

4. The method of claim 3, wherein the protecting group of the alcohol is an acetonide, a silyl protecting group, an alkyl protecting group, or an aryl protecting group.

5. The method of claim 3, wherein the protecting group of the alcohol is tert-butyl(dimethyl)silyl (TBS), triisopropylsilyl (TIPS), trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), triethylsilyl (TES), 2-(trimethylsilyl)ethoxymethyl ether (SEM), tetrahydropyranyl (THP) ether, acetate (Ac), pivalate (Piv), methoxymethyl, or p-methoxybenzyl (PMB).

6. The aforementioned halohydrin ketone compound, It is one of the following: During the ceremony, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, and also, The method of claim 1, wherein X is a halogen.

7. The aforementioned halohydrin ketone compound, It is one of the following: During the ceremony, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, X is a halogen, and also Y is CH 2 The method of claim 1, wherein O, S, and NR are the components, and R is an alkyl, aryl, acyl, or carboxylate.

8. The aforementioned halohydrin ketone compound, The method according to claim 1.

9. The method according to claim 1, i) Halohydrin ketone compounds with the following structure: (In the formula, X is a halogen, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, and also, To provide a product in which PG is a protecting group, and ii) The halohydrin ketone compound is reacted with a soft nucleophile to obtain a nucleoside analog in which the C4' position of the following structure is substituted: (In the formula, NB is an optionally substituted aryl, arylalkyl, heteroaryl, or heteroarylalkyl, Nu is a soft nucleophile, and also To generate a protecting group in which PG can arbitrarily form a covalent bond with Nu. Methods that include...

10. The method of claim 9, wherein the protecting group is an alcohol protecting group.

11. The method of claim 10, wherein the protecting group of the alcohol is an acetonide, a silyl protecting group, an alkyl protecting group, or an aryl protecting group.

12. The method of claim 10, wherein the protecting group of the alcohol is tert-butyl(dimethyl)silyl (TBS), triisopropylsilyl (TIPS), trimethylsilyl (TMS), tert-butyldiphenylsilyl (TBDPS), triethylsilyl (TES), 2-(trimethylsilyl)ethoxymethyl ether (SEM), tetrahydropyranyl (THP) ether, acetate (Ac), pivalate (Piv), methoxymethyl, or p-methoxybenzyl (PMB), and is not limited to acetonides, silyl protecting groups, alkyl protecting groups, or aryl protecting groups (including cyclic or acyclic).

13. The soft nucleophile is MeCN, N 3 - 5CN - , or MeNO 2 The method according to any one of claims 1 to 12.

14. The method according to any one of claims 1 to 13, wherein the halogen is F.

15. NB includes phenyl, biphenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, dihydrobenzopyranyl, 1,4-benzodioxanyl, furan, thiophene, pyrrole, oxazole, thiazole, imidazole, pyrazole, isoxazole, isothiazole, 1,2,3-oxadiazole, triazole, 1,3,4-thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, 2,6-dichloropyrimidinepyrazine, 1,3,5-triazine, imidazole, benzimidazole, benzoxazole, benzothiazole, indidine, in The method according to any one of claims 1 to 14, wherein the material is dole, isoindole, benzofuran, benzothiophene, 1H-indazole, purine, 4H-quinolidine, quinoline, isoquinoline, cinnoline, phthalazine, quinazoline, quinoxaline, 1,8-naphthyridine, pteridine, uracil, thymine, deazadenine, phthalimide, adenine, 5,6-dihydrouracil, 5-methylcytosine, 5-hydroxymethylcytosine, 5,5,5-trifluoromethylthymine, 5-fluorouracil, 2-thiouracil, 4-methylbenzimidazole, hypoxanthine, 7-deazaguanine, or 7-deazaadenine.

16. The method according to any one of claims 1 to 15, wherein the nucleoside analog is a monomer in a loc nucleic acid (LNA), a small molecule therapeutic agent, or an oligonucleotide therapeutic agent.

17. The method according to claim 1, wherein the nucleoside analog comprises a nitrile moiety and can be reduced to an aldehyde.

18. The nucleoside analog in which the C4' position is substituted is The method according to claim 1. Claim 19: A nucleoside analog in which the C4' position is substituted.

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