Reagents for the modular enantiodivergent synthesis of CP bonds and their uses

Abstract

This disclosure describes chiral P(V)-based reagents and their use for the modular, scalable, and stereospecific synthesis of chiral phosphines, phosphine oxides, and specific oligonucleotides.

Description

[Technical Field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 962,066, filed January 16, 2020. [Background technology]

[0002] Controlling the stereochemistry of P centers is a common challenge encountered in a variety of fields. See Corbridge, DEC, Phosphorus: Chemistry, Biochemistry and Technology, 6th ed.; Taylor & Francis, 2013. A recent example comes from the emergence of antisense oligonucleotide (ASO) therapy, where precise control of the P stereocenter in phosphorothioate linkages can affect efficacy and physical properties. See Bohr, HG et al., "Electronic Structures of LNA Phosphorothioate Oligonucleotides," Mol. Ther. Nucleic Acids 2017, 8, 428-441; see also Iwamoto, N., "Control of Phosphorothioate Stereochemistry Substantially Increases the Efficacy of Antisense Oligonucleotides," Nat. Biotechnol. 2017, 35(9), 845-851. U.S. Patent Application Publication No. 2019 / 0322694 described a method for precisely controlling the stereochemical outcome of such systems (phosphorus-sulfur incorporation, or Ψ for short) using P(V)-based reagents built on the inexpensive limonene scaffold. The advantage of such a method is that, unlike classical phosphoramidite P(III)-based reagents, Ψ-enabled ASO synthesis is redox-economical and involves air- and moisture-tolerant building blocks.

[0003] Another P-based chiral linkage that is gaining interest is found in methyl-phosphonate oligonucleotides, or MPOs. See Pritchard, C.E. et al., "Methylphosphonate mapping of phosphate contacts critical for RNA recognition by the human immunodeficiency virus tat and rev proteins," Nucleic Acids Res. 1994, 22(13), 2592-2600; see also Koch, M. et al., "Role of a ribosomal RNA phosphate oxygen during the EF-G-triggered GTP hydrolysis," Proc. Natl. Acad. Sci. 2015, 112(20), E2561; Hamma, T. et al., "Interactions of Hairpin Oligo-2'-O-Methylribonucleotides Containing Methylphosphonate Linkages with HIV TAR RNA," Antisense and Nucleic Acid Drug Dev. 2003, 13(1), 19-30. Such structures are thought to have superior stability due to their greater resistance to phosphodiesterase degradation. See Agrawal, S. et al., "Oligodeoxynucleoside Methylphosphonates: Synthesis and Enzymic Degradation," Tetrahedron Lett. 1987, 28(31), 3539-3542; Reynolds, M.A. et al., "Synthesis and Thermodynamics of Oligonucleotides Containing Chirally Pure RP Methylphosphonate Linkages," Nucleic Acids Res. 1996, 24(22), 4584-4591. Due to the absence of bridging oxygen atoms, the backbone is charge-neutral and incapable of H-bonding.This can have either a stabilizing or destabilizing effect on the formation of DNA-RNA duplexes. Similar to phosphorothioate-based ASOs, MPO introduces chirality at the phosphorus, resulting in S. p and R p The isomers of α-ASO have different binding affinities and biological activities. Indeed, one of the first ASO analogs utilized MPO linkage. Currently, the preparation of MPO-based building blocks is coupled to P(III)-based methods, which require laborious separation of diastereomers. Miller, PS et al., Nonionic nucleic acid analogs. Synthesis and characterization of dideoxyribonucleoside methylphosphonates. Biochemistry 1979, 18(23), 5134-5143; Engels, J. et al., Eine neue Synthese von Nukleosidmethylphosphonaten. Angew. Chem. Int. Ed. Eng. 1982, 21(S12), 2010-2015; Lebedev, AV et al., "A new DMAP-catalyzed phosphonamidite coupling reaction for synthesis of oligonucleotide methylphosphonate derivatives" Tetrahedron Lett. 1990, 31(6), 851-854; Helinski, J. et al., “N,N-diisopropyl-OP-nitrophenyl-P-methylphosphonoamidite: Novel Difunctional PIII Reagent in Oligonucleoside Methylphosphonate Synthesis Containing See "4-Nitrophenoxy Group" Tetrahedron Lett. 1991, 32(37), 4981-4984. These limitations have certainly prevented widespread exploration of such linkages for modern therapeutic applications.

[0004] In addition, stereogenic P-based systems that are chiral at phosphorus represent some of the most important ligands known. See Vineyard, B.D. et al., Asymmetric Hydrogenation: Rhodium Chiral Bisphosphine Catalyst, J. Am. Chem. Soc. 1977, 99(18), 5946-5952; see also Knowles, W.S., "Asymmetric Hydrogenations (Nobel Lecture 2001)," Adv. Synth. Catal. 2003, 345(1-2), 3-13. For example, DIPAMP was the first chiral phosphine utilized in asymmetric hydrogenation and played a central role in the legendary L-DOPA process. Since then, numerous useful ligand scaffolds have emerged, including DuanPhos and BIBOP, which allow the introduction of new stereogenic carbon centers with nearly complete control. Despite their great utility, a facile, modular synthesis of P-chiral phosphines remains an unsolved challenge.

[0005] Thus, there is a need for efficient and straightforward methods to control P-based stereochemistry in applications including oligonucleotide chemistry and asymmetric synthesis. Summary of the Invention

[0006] In one aspect, the present disclosure provides a compound of formula (I): [ka] [In the formula: R 1 , R 4 , R 5 , R 7 , and n is as defined below] or a salt thereof.

[0007] In one aspect, the present disclosure provides a compound of formula (Ia): [ka] The present invention relates to a compound of the formula:

[0008] In one aspect, the disclosure provides a method for preparing a thiophosphinic acid, comprising reacting a carbanion reagent with a compound of formula (Ia): [ka] [In the formula: R 1 is defined below] or a salt thereof, an enantiomer thereof, or a combination thereof to form a thiophosphinic acid.

[0009] In one embodiment, the thiophosphinic acid formed in the above method has the formula (II): [ka] [In the formula: R 1 and R 2 is defined below] It is expressed as:

[0010] In one embodiment, the method further comprises reacting the thiophosphinic acid with an alkylating reagent to form a thiophosphinic ester.

[0011] In one embodiment, the thiophosphinate ester formed in the above process has the formula (III): [ka] [In the formula: R 1 , R 2 and R' is as defined below. It is expressed as:

[0012] In one embodiment, the method further comprises reacting the thiophosphinate ester with an alkoxide agent to form an organophosphinate.

[0013] In one embodiment, the organophosphinate formed in the above method has the formula (IV): [ka] [In the formula: R 1 , R 2 and R' is as defined below. It is expressed as:

[0014] In one embodiment, the method further comprises reacting the organophosphinate with a carbanion reagent to form a phosphine oxide.

[0015] In one embodiment, the phosphine oxide formed in the above process has the formula (V): [ka] [In the formula: R 1 , R 2 , and R 3 is defined below] It is expressed as:

[0016] In one embodiment, the method further comprises reacting the phosphine oxide with a reducing agent to form a phosphine.

[0017] In one embodiment, the phosphine formed in the above process has the formula (VI): [ka] [In the formula: R 1 , R 2 , and R 3 is defined below] It is expressed as:

[0018] In one aspect, the present disclosure relates to a method for preparing a nucleoside-loaded organophosphorus compound, comprising reacting a compound of formula (Ia) or an enantiomer thereof with a nucleoside.

[0019] In one embodiment, the nucleoside-loaded organophosphorus compound formed in the above method has the formula (VII) or (VIIa): [ka] [In the formula: R 1 , R', or Nu 1 is defined below] or a diastereomer thereof.

[0020] In one embodiment, the method further comprises reacting the nucleoside-loaded organophosphorus compound with a second nucleoside to form a dinucleotide.

[0021] In one embodiment, the dinucleotide formed in the above method has formula (VIII) or (VIIIa): [ka] [In the formula: R 1 , Nu 1 , or Nu 2 is defined below] It is expressed as:

[0022] In one embodiment, the method comprises: a) reacting the dinucleotide with a compound of formula (Ia) to form a loaded dinucleotide; b) reacting the loaded dinucleotide formed in step (a) with a third nucleoside to form a trinucleotide; c) repeating steps (a) and (b) one or more times to form an oligonucleotide having a desired number of nucleotides. It further includes:

[0023] In one embodiment, the method comprises: a) Reconstituting a dinucleotide with formula C: [ka] or an enantiomer thereof to form a loaded chimeric dinucleotide; and b) reacting the loaded chimeric dinucleotide formed in step (a) with a third nucleoside to form a chimeric trinucleotide having phosphonate and phosphorothioate linkages; It further includes:

[0024] In one aspect, the present disclosure provides a compound of formula (IIIa): [ka] [In the formula: R 1 and R 2 is defined below] or an enantiomer thereof.

[0025] In one embodiment, the present disclosure provides a compound of formula (IVa): [ka] [In the formula: R 1 and R 2 is defined below] or an enantiomer thereof.

[0026] In one embodiment, the present disclosure provides a compound of formula (VII): [ka] [In the formula: R 1 , R', and Nu 1 is defined below] or an enantiomer thereof.

[0027] In one aspect, the present disclosure provides a compound of formula (Ia): [ka] [In the formula: R 1 is defined below] or an enantiomer thereof, comprising: The carbanion reagent is reacted with a compound of Formula A: [ka] or an enantiomer thereof to form a compound of formula (Ia). DETAILED DESCRIPTION OF THE INVENTION

[0028] Disclosed Reagents The disclosed reagents and compounds are P(V)-based compounds for the modular enantiodivergent synthesis of CP bonds.

[0029] a. Phosphonodithioate reagents In one embodiment, the phosphonodithioate reagent has the formula (I): [ka] [In formula: R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl, where R 1 optionally, one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; R 4 is hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; R 5 is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; R 7 is a substituent that replaces hydrogen, and C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, or C 6-10 aryl; and n is 0, 1, 2, 3, 4, 5, 6, 7, or 8] or a salt thereof.

[0030] In one embodiment, the phosphonodithioate reagent has the formula (Ia): [ka] [In the formula: R 1 is as defined above] or a salt thereof, an enantiomer thereof, or a combination thereof.

[0031] In one embodiment, R 1 teeth, [ka] selected from the group consisting of

[0032] where [ka] is R 1 and the phosphorus of formula (I).

[0033] In one embodiment, R 1 is -CH3. In another embodiment, R 1 is phenyl.

[0034] In one embodiment, the phosphonodithioate reagent is any one or more of the compounds listed in Table 1. Table 1 [Table 1] [Table 2] [Table 3]

[0035] In one embodiment, the phosphonodithioate reagents of the disclosure have an optical purity or enantiomeric excess (as measured by standard methods in the art) of at least 50% (i.e., at least 75% of one enantiomer and up to 25% of the other enantiomer), at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0036] b. Thiophosphinate Reagents In another aspect, the present disclosure provides a compound of formula (IIIa): [ka] [In formula: R 1 and R 2 are respectively, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R 1 and R 2 each optionally containing one or more of the same or different R a may be substituted with a group; R ais deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] or an enantiomer thereof.

[0037] In one embodiment, the thiophosphinate reagent is any one or more of the compounds listed in Table 2. [Table 4] [Table 5]

[0038] In one embodiment, the thiophosphinate ester reagents of the present disclosure have an optical purity or enantiomeric excess (as measured by standard methods in the art) of at least 50% (i.e., at least 75% of one enantiomer and up to 25% of the other enantiomer), at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0039] c. Organic phosphinate reagents In another aspect, the present disclosure provides a compound of formula (IVa): [ka] [In formula: R 1 and R 2 are respectively, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R 1 and R 2 each optionally containing one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] or an enantiomer thereof.

[0040] In one embodiment, the organophosphinate reagent is any one or more of the compounds listed in Table 3. Table 3 [Table 6]

[0041] In one embodiment, the organophosphinate reagents of the present disclosure have an optical purity or enantiomeric excess (as measured by standard methods in the art) of at least 50% (i.e., at least 75% of one enantiomer and up to 25% of the other enantiomer), at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0042] d. Nucleoside-loaded organophosphorus reagents In one embodiment, the present disclosure provides a compound of formula (VII) or (VIIa): [ka] [In formula: R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; R' is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; Nu 1 is a nucleoside, Here, Nu 1 may optionally contain, independently of one another, one or more of the same or different modifications or one or more of the same or different protecting groups. The present invention provides a nucleoside-loaded organophosphorus reagent represented by the formula:

[0043] In one embodiment, R 1 and R' are each -CH3.

[0044] In one embodiment, Nu 1 teeth, [ka] A nucleoside selected from the group consisting of: T is [ka] and; A Bz teeth [ka] and; G iBu teeth [ka] and C Bz teeth [ka] is a nucleoside.

[0045] In one embodiment, the nucleoside-loaded organophosphorus reagent is any one or more of the compounds listed in Table 4. Table 4 [Table 7]

[0046] In one embodiment, the nucleoside-loaded organophosphorus reagents of the present disclosure have a diastereomeric ratio ("dr") of at least 5:1, at least 10:1, at least 15:1, at least 20:1, at least 25:1, at least 30:1, at least 35:1, at least 40:1, at least 45:1, at least 50:1, at least 55:1, at least 60:1, at least 65:1, at least 70:1, at least 75:1, at least 80:1, at least 85:1, at least 90:1, at least 95:1, or at least 99:1 (as measured by standard methods in the art).

[0047] In one embodiment, the nucleoside is a ribonucleoside. In another embodiment, the nucleoside is a deoxyribonucleoside.

[0048] Nucleosides can be naturally occurring nucleosides or non-naturally occurring nucleoside analogs. As used herein, "nucleoside analogs" refer to variants of natural nucleosides, such as DNA or RNA nucleosides, modified by modifying sugar and / or base moieties. In the context of oligonucleotides, analogs can be essentially "silent" or "equivalent" to natural nucleosides, i.e., they do not exert a functional effect during the function of the oligonucleotide. Such "equivalent" analogs can still be useful, for example, if they are easier to produce, cheaper, more stable under storage or manufacturing conditions, or represent tags or labels. However, in some embodiments, analogs will have a functional effect during the function of the oligonucleotide, for example, by increasing the binding affinity to the target, and / or increasing resistance to intracellular nucleases, and / or increasing the ease of transport into cells.

[0049] Nucleosides useful herein can also contain modified sugars. Modifications of the 2'-sugar include fluoro, O-alkyl, O-alkylamino, O-alkylalkoxy, protected O-alkylamino, O-alkylaminoalkyl, O-alkylimidazole, and groups of the formula (O-alkyl) m(wherein m is 1 to about 10). Preferred among these polyethers are linear or cyclic polyethylene glycols (PEGs) and PEG-containing groups, such as crown ethers and the polyethers disclosed in Ouchi et al., Drug Design and Discovery 1992, 9, 93; Ravasio et al., J. Org. Chem. 1991, 56, 4329; and Delgardo et al., Critical Reviews in Therapeutic Drug Carrier Systems 1992, 9, 249 (each of which is incorporated herein by reference in its entirety). Additional nucleosides embodying sugar modifications are disclosed in Cook, Anti-Cancer Drug Design, 1991, 6, 585-607 and U.S. Publication No. 2016 / 237427 (incorporated herein by reference in its entirety). Fluoro, O-alkyl, O-alkylamino, O-alkylimidazole, O-alkylaminoalkyl, and alkylamino substitutions are described in U.S. Pat. No. 6,166,197, entitled "Oligomeric Compounds Having 2'- and 5'-Substituted Pyrimidine Nucleotides," which is incorporated herein by reference in its entirety.

[0050] Further useful nucleosides with 2'-sugar modifications include 2'-SR and 2'-NR groups, where each R is independently hydrogen, a protecting group, or a substituted or unsubstituted alkyl, alkenyl, or alkynyl. 2'-SR nucleosides are disclosed in U.S. Patent No. 5,670,633, issued September 23, 1997, which is incorporated herein by reference in its entirety.

[0051] Useful nucleosides also include nucleosides that are derivatized with selenium (Se). Examples of Se-derivatized nucleosides include nucleosides in which the 2' and / or 5' O-atoms of the sugar are replaced with Se. Other examples include nucleosides in which the oxygen in the furanose ring is replaced with Se, nucleobases, and non-bridging phosphates. Such nucleic acids are described, for example, in Pallan et al., Nat. Protoc., 2(3): 647-51 (2007) and Nat. Protoc., 2(3); 640-646 (2007) (incorporated herein in their entirety).

[0052] Other examples of suitable nucleosides include those described in Schinazi et al., Nucleosides and Nucleotides, 17: 635-647 (1998); Biochem., 35(18): 5741-5746 (1996); J. Org. Chem., 79(8): 3465-3472 (2014), which are incorporated herein by reference in their entireties.

[0053] Further useful nucleoside analogs include, but are not limited to, locked nucleic acids (LNA); 2'-O-alkyl-RNA; 2'-amino-DNA; 2'-fluoro-DNA; arabinonucleic acid (ANA); 2'-fluoro-ANA, hexitol nucleic acid (HNA), intercalating nucleic acid (INA), constrained ethyl nucleosides (cEt), 2'-O-methyl nucleic acid (2'-OMe), 2'-O-methoxyethyl nucleic acid (2'-MOE), or any combination thereof.

[0054] "Hexitol nucleic acid" or "HNA" is composed of phosphorylated 2,3-dideoxy-D-arabino-hexitol units in which the nucleobase is located at the 2-[S]-position.

[0055] "cEt" or "constrained ethyl" means a bicyclic nucleoside having a sugar moiety containing a bridge connecting the 4'-carbon and the 2'-carbon, where the bridge is represented by the formula: 4'-CH(CH3)-0-2'.

[0056] "2'-O-Methoxyethyl" (also referred to as 2'-MOE, and 2'-O(CH2)2-OCH3, and MOE) refers to an O-methoxy-ethyl modification at the 2' position of the furanosyl ring. A 2'-O-methoxyethyl modified sugar is a modified sugar.

[0057] "2'-F" refers to a modification at the 2' position of the furanosyl sugar ring to include a fluoro group.

[0058] As used herein, "2'-OMe" or "2'-OCH3" or "2'-O-methyl" each refer to a modification at the 2' position of the furanosyl sugar ring to include an -OCH3 group.

[0059] Examples of suitable nucleotide analogues are provided by or referenced in WO2007 / 031091, which is incorporated by reference in its entirety.

[0060] Incorporation of affinity-enhancing nucleotide analogs, such as LNA or 2'-substituted sugars, into oligomers can reduce the size of oligomers that specifically bind, even lowering the upper limit to the size of oligomers before nonspecific or aberrant binding occurs.

[0061] In some embodiments, the nucleoside is a nucleoside analog containing a bicyclic sugar. Non-limiting exemplary bicyclic sugars include cEt, 2',4'-constrained 2'-O-methoxyethyl (cMOE), LNA, α-LNA, β-LNA, 2'-O,4'-C-ethylene-bridged nucleic acid (ENA), amino-LNA, oxy-LNA, or thio-LNA.

[0062] The term "LNA" refers to a bicyclic nucleoside analog known as a "locked nucleic acid." It can refer to an LNA monomer, or, when used in the context of "LNA oligonucleotides," LNA refers to an oligonucleotide containing one or more such bicyclic nucleotide analogs. LNA nucleosides are characterized by the presence of a linker group (e.g., a bridge) between C2' and C4' of the ribose sugar ring. The bridge includes, but is not limited to, a biradical selected from -CH2-O-, -CH2-S-, -CH2-NH-, -CH2-N(CH3)-, -CH2-CH2-O-, -CH2-CH(CH3)-, -CH2-CH2-S-, -CH2-CH2-NH-, -CH2-CH2-CH2-, -CH2-CH2-CH2-O-, -CH2-CH2-CH(CH3)-, -CH=CH-CH2-, -CH2-O-CH2-O-, -CH2-NH-O-, -CH2-N(CH3)-O-, -CH2-O-CH2-, -CH(CH3)-O-, and -CH(CH2-O-CH3)-O-, and / or -CH2-CH2- and -CH=CH-. For all chiral centers, the asymmetric groups can be found in either the R or S orientation.

[0063] In some embodiments, the biradical is —C(R a R b )-OC(R c R d )-O-, where R a , R b , R c , and R d is hydrogen, halogen, C 1-6 Alkyl, substituted C 1-6 Alkyl, C 2-6 Alkenyl, substituted C 2-6 Alkenyl, C 2-6 Alkynyl or substituted C 2-6 Alkynyl, C 1-6 Alkoxyl, substituted C 1-6 Alkoxyl, acyl, substituted acyl, C 1-6 Aminoalkyl or substituted C 1-6 and independently selected from the group consisting of aminoalkyl, for example hydrogen.

[0064] Suitable bicyclic nucleosides are disclosed in WO2007 / 134181, WO2008 / 154401, WO2008 / 150729, WO2009 / 067647 (alpha-L-bicyclic nucleic acid analogs) and WO2009006478A, all of which are incorporated herein by reference in their entirety.

[0065] Additional bicyclic nucleoside analogs and their use in antisense oligonucleotides are disclosed in WO2011 / 115818, WO2011 / 085102, WO2011 / 017521, WO2009 / 100320, WO2010 / 036698, WO2009 / 124295, and WO2009 / 006478A, each of which is incorporated herein by reference in its entirety.

[0066] The term "thio-LNA" includes locked nucleosides in which Y in the general formula III below is selected from S or -CH2-S-. Thio-LNA can be in both the beta-D and alpha-L-configuration.

[0067] The term "amino-LNA" refers to the amino group in the general formula III below, where Y is -N(H)-, -N(R)-, -CH-N(H)-, and -CH-N(R)- (where R is hydrogen and C 1-4 Amino-LNA can be in both beta-D and alpha-L-configuration.

[0068] The term "oxy-LNA" includes locked nucleosides in which Y represents -O- in the general formula III below. Oxy-LNA can be in both the beta-D and alpha-L-configuration.

[0069] The term "ENA" includes locked nucleosides having the following general formula: Y is -CH2-O- (where the oxygen atom of -CH2-O- is attached in the 2' position relative to the base B): R e is hydrogen or methyl.

[0070] In some exemplary embodiments, the LNA is selected from beta-D-oxy-LNA, alpha-L-oxy-LNA, beta-D-amino-LNA, and beta-D-thio-LNA.

[0071] In some embodiments, the nucleoside analog may be, for example: a 2'-O-alkyl-RNA unit, a 2'-amino-DNA unit, a 2'-fluoro-DNA unit, an LNA unit, an arabinonucleic acid (ANA) unit, a 2'-fluoro-ANA unit, an HNA unit, an INA (intercalating nucleic acid - Christensen, 2002. Nucl. Acids. Res. 2002 30:4918-4925, incorporated herein by reference) unit, and a 2'-MOE unit.

[0072] The term "nucleobase" refers to the base portion of a nucleotide and encompasses both naturally occurring and non-naturally occurring variants. Thus, "nucleobase" encompasses not only the known purine and pyrimidine-type heterocycles, but also heterocyclic analogs and tautomers thereof.

[0073] Exemplary nucleobases include, but are not limited to, adenine, guanine, cytosine, thymidine, uracil, xanthine, hypoxanthine, 5-methylcytosine, isocytosine, pseudoisocytosine, 5-bromouracil, 5-propynyluracil, 6-aminopurine, 2-aminopurine, inosine, diaminopurine, and 2-chloro-6-aminopurine.

[0074] In some embodiments, nucleosides include naturally occurring nucleobases such as adenine, guanine, cytosine, uridine, thymine, 5-methylcytosine, etc. In other embodiments, nucleosides include other natural nucleobases as well as modified nucleobases such as xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 5-halo uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, oxa, amino, thiol, thioalkyl, hydroxyl and other 8-substituted adenines and guanines, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine, etc. Further, naturally occurring and non-naturally occurring nucleobases include those described in U.S. Pat. No. 3,687,808 (Merigan et al.); Sanghvi, in Antisense Research and Application, Chapter 15, edited by S.T. Crooke and B. Lebleu, CRC Press, 1993; Englisch et al., Angewandte Chemie, International Edition, 1991, 30, 613-722 (especially pages 622 and 623); the Concise Encyclopedia of Polymer Science and Engineering, edited by J.I. Kroschwitz, John Wiley & Sons, 1990, pp. 858-859; Zhang et al., Nature, 2017, 551, 644-647 (hydrophobic bases); Feldman and Romesberg, Acc. Chem., Res., 2018, 51, 394-403; and Cook, Anti-Cancer Drug Design, 1991, 6, 585-607, each of which is incorporated herein by reference in its entirety.

[0075] Other examples of modified nucleosides and nucleobases described herein include, but are not limited to, the following modifications: 2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine; 2-methylthio-N6-methyladenosine; 2-methylthio-N6-threonylcarbamoyladenosine; N6-glycinylcarbamoyladenosine; N6-isopentenyladenosine; N6-methyladenosine; N6-threonylcarbamoyladenosine; 1,2'-O-dimethyladenosine; 1-methyladenosine; 2'-O-methyladenosine; 2'-O-ribosyladenosine (phosphate); 2-methyladenosine; 2-methylthio-N6-isopentenyladenosine; 2-methylthio-N6-hydroxynorvalyl. Carbamoyl adenosine;2'-O-methyl adenosine;2'-O-ribosyladenosine (phosphate);Isopentenyladenosine;N6-(cis-hydroxyisopentenyl)adenosine;N6,2'-O-dimethyladenosine;N6,2'-O-dimethyladenosine;N6,N6,2'-O-trimethyladenosine;N6,N6-dimethyladenosine;N6-acetyladenosine;N6-hydroxynorvalylcarbamoyl adenosine;N6-methyl-N6-threonylcarbamoyl adenosine;2-methylthio-N 6-Isopentenyladenosine;7-Deaza-adenosine;N1-Methyl-adenosine;N6,N6-(Dimethyl)adenine;N6-cis-Hydroxy-isopentenyl-adenosine;α-Thio-adenosine;2-(Amino)adenine;2-(Aminopropyl)adenine;2-(Methylthio)-N6-(isopentenyl)adenine;2-(Alkyl)adenines;2-(Aminoalkyl)adenines;2-(Aminopropyl)adenine;2-(Halo)adenine;2-(Propyl)adenine;2'-Amino-2'-deoxyadenosine Triphosphate;2'-Azido-2'-deoxy-adenosine triphosphate;2'-Deoxy-2'-α-aminoadenosine triphosphate;2'-Deoxy-2'-α-azidoadenosine triphosphate;6-(Alkyl)adenine;6-(Methyl)adenine;7-(Deaza)adenine;8-(Alkynyl)adenine;8-(Alkenyl)adenine;8-(Alkyl)adenine;8-(Alkynyl)adenine8-(Amino)adenine;8-(Halo)adenine;8-(Hydroxyl)adenine;8-(Thioalkyl)adenine;8-(Thiolate)adenine;8-Azido-adenosine;Azaadenine;Deazaadenine;N6-(Methyl)adenine;N6-(Isopentyl)adenine;7-Deaza-8-aza-adenosine;7-Methyladenine;1-Deazaadenosine triphosphate;2'-Fluoro-N6-Bz-deoxyadenosine triphosphate;2'-Methoxy-2-amino-adenosine triphosphate;2'O-Methyl-N6-Bz-deoxyadenosine triphosphate;2'-α-Ethynyladenosine triphosphate;2-Aminoadenine;2-Aminoadenosine triphosphate Triphosphate;2'-a-Trifluoromethyladenosine triphosphate;2-Azidoadenosine triphosphate;2'-b-Ethynyladenosine triphosphate;2-Bromoadenosine triphosphate;2'-b-Trifluoromethyladenosine triphosphate;2-Chloroadenosine triphosphate;2'-Deoxy-2',2'-difluoroadenosine triphosphate;2'-Deoxy-2'-a-mercaptoadenosine triphosphate;2'-Deoxy-2'-a-thiomethoxyadenosine triphosphate;2'-Deoxy-2'-b-aminoadenosine triphosphate;2'-Deoxy-2'-b-azidoadenosine triphosphate;2'-Deoxy-2'-b-bromoadenosine Triphosphate;2'-Deoxy-2'-b-chloroadenosine triphosphate;2'-Deoxy-2'-b-fluoroadenosine triphosphate;2'-Deoxy-2'-b-iodoadenosine triphosphate;2'-Deoxy-2'-b-mercaptoadenosine triphosphate;2'-Deoxy-2'-b-thiomethoxyadenosine triphosphate;2-Fluoroadenosine triphosphate;2-Iodoadenosine triphosphate;2-Mercaptoadenosine triphosphate;2-Methoxy-adenine;2-Methylthio-adenine;2-Trifluoromethyladenosine triphosphate;3-Deaza-3-bromoadenosine triphosphate;3-Deaza-3-chloroadenosine triphosphate;3-Deaza-3-fluoroadenosine triphosphate;3-Deaza-3-iodoadenosine triphosphate;3-Deazaadenosine triphosphate;4'-Azidoadenosine triphosphate;4'-Carbocyclic adenosine triphosphate;4'-Ethynyladenosine triphosphate;5'-Homoadenosine triphosphate;8-Aza-adenosine triphosphate;8-Bromoadenosine triphosphate;8-Trifluoromethyladenosine triphosphate;9-Deazaadenosine Triphosphate;2-aminopurine;7-deaza-2,6-diaminopurine;7-deaza-8-aza-2,6-diaminopurine;7-deaza-8-aza-2-aminopurine;2,6-diaminopurine;7-deaza-8-aza-adenine, 7-deaza-2-aminopurine;2-thiocytidine;3-methylcytidine;5-formylcytidine;5-hydroxymethylcytidine;5-methylcytidine;N4-acetylcytidine;2'-O-methylcytidine;5,2'-O-dimethylcytidine;5-formyl-2'-O-methylcytidine;lysidine;N4,2'-O-dimethylcytidine;N4-acetyl-2'-O-methylcytidine;N4-methylcytidine;N4,N4-dimethyl-2'-OMe-cytidine TP; 4-methylcytidine; 5-aza-cytidine; pseudoisocytidine; pyrrolo-cytidine; α-thio-cytidine; 2-(thio)cytosine; 2'-amino-2'-deoxy-cytidine triphosphate; 2'-azido-2'-deoxy-cytidine triphosphate; 2'-deoxy-2'-α-aminocytidine triphosphate; 2'-deoxy-2'-α-azidocytidine Triphosphate;3(deaza)-5-(aza)cytosine;3(methyl)cytosine;3-(alkyl)cytosine;3-(deaza)-5-(aza)cytosine;3-(methyl)cytidine;4,2'-O-dimethylcytidine;5-(halo)cytosine;5-(methyl)cytosine;5-(propynyl)cytosine;5-(trifluoromethyl)cytosine;5-(alkyl)cytosine;5-(alkynyl)cytosine;5-(halo)cytosine;5-(propynyl)cytosine;5-(trifluoromethyl)cytosine;5-bromo-cytidine;5-iodo-cytidine;

[0076] 5-Propynylcytosine;6-(Azo)cytosine;6-Aza-cytidine;Azacytosine;Deazacytosine;N4-(Acetyl)cytosine;1-Methyl-1-deaza-pseudoisocytidine;1-Methyl-pseudoisocytidine;2-Methoxy-5-methyl-cytidine;2-Methoxy-cytidine;2-Thio-5-methyl-cytidine;4-Methoxy-1-methyl-pseudoisocytidine;4-Methoxy-pseudoisocytidine;4-Thio-1-methyl-1-deaza-pseudoisocytidine;4-Thio-1-methyl-pseudoisocytidine;4-Thio-pseudoisocytidine;5-Aza-zebularine;5-Methyl-zebularine;Pyrrolo-pseudoisocytidine;Zebularine;(E)-5-(2-Bromo-vinyl)cytidine Triphosphate;2,2'-Anhydro-cytidine triphosphate hydrochloride;2'-Fluoro-N4-Bz-cytidine triphosphate;2'-Fluoro-N4-acetyl-cytidine triphosphate;2'-O-Methyl-N4-acetyl-cytidine triphosphate;2'O-Methyl-N4-Bz-cytidine triphosphate;2'-a-Ethynylcytidine triphosphate;2'-a-Trifluoromethylcytidine triphosphate;2'-b-Ethynylcytidine triphosphate;2'-b-Trifluoromethylcytidine triphosphate;2'-Deoxy-2',2'-difluorocytidine triphosphate;2'-Deoxy-2'-a-mercaptocytidine triphosphate;2'-Deoxy-2'-a-thiomethoxycytidine Triphosphate;2'-Deoxy-2'-b-aminocytidine triphosphate;2'-Deoxy-2'-b-azidocytidine triphosphate;2'-Deoxy-2'-b-bromocytidine triphosphate;2'-Deoxy-2'-b-chlorocytidine triphosphate;2'-Deoxy-2'-b-fluorocytidine triphosphate;2'-Deoxy-2'-b-iodocytidine triphosphate;2'-Deoxy-2'-b-mercaptocytidine triphosphate;2'-Deoxy-2'-b-thiomethoxycytidine triphosphate;2'-O-Methyl-5-(1-propynyl)cytidine triphosphate;3'-Ethynylcytidine triphosphate;4'-Azidocytidine triphosphate;4'-Carbocyclic cytidine triphosphate;4'-Ethynylcytidine triphosphate;5-(1-Propynyl)-aracytidine triphosphate;5-(2-Chloro-phenyl)-2-thiocytidine triphosphate;5-(4-Amino-phenyl)-2-thiocytidine triphosphate;5-Aminoallyl-cytidine triphosphate;5-Cyanocytidine triphosphate;5-Ethynyl-aracytidine triphosphate;5-Ethynylcytidine triphosphate;5'-Homocytidine triphosphate;5-Methoxycytidine triphosphate;5-Trifluoromethyl-cytidine triphosphate;N4-Amino-cytidine triphosphate;N4-Benzoyl-cytidine Triphosphate;Pseudoisocytidine;7-Methylguanosine;N2,2'-O-Dimethylguanosine;N2-Methylguanosine;Wyosine;1,2'-O-Dimethylguanosine;1-Methylguanosine;2'-O-Methylguanosine;2'-O-Ribosylguanosine(phosphate);2'-O-Methylguanosine;2'-O-Ribosylguanosine(phosphate);7-Aminomethyl-7-deazaguanosine;7-Cyano-7-deazaguanosine;Archae Osine;Methylwosine;N2,7-Dimethylguanosine;N2,N2,2'-O-Trimethylguanosine;N2,N2,7-Trimethylguanosine;N2,N2-Dimethylguanosine;N2,7,2'-O-Trimethylguanosine;6-Thio-guanosine;7-Deaza-guanosine;8-Oxo-guanosine;N1-Methyl-guanosine;α-Thio-guanosine;2(Propyl)guanine;2-(Alkyl)guanine;2'-Amino-2'-deoxy-guanosine Triphosphate;2'-Azido-2'-deoxy-guanosine triphosphate;2'-Deoxy-2'-α-aminoguanosine triphosphate;2'-Deoxy-2'-α-azidoguanosine triphosphate;6-(Alkyl)guanine;;6-Methyl-guanosine;7-(Alkyl)guanine;7-(Deaza)guanine;7-(Methyl)guanine;8-(Alkenyl)guanine;8-(Alkyl)guanine;8-(Alkynyl)guanine;8-(Amino)guanine;8-(Halo)guanine8-(Hydroxyl)guanine;8-(Thioalkyl)guanine;8-(Thiol)guanine;Azaguanine;Deazaguanine;N-(Methyl)guanine;1-Methyl-6-thio-guanosine;6-Methoxy-guanosine;6-Thio-7-deaza-8-aza-guanosine;6-Thio-7-deaza-guanosine;6-Thio-7-methyl-guanosine;7-Deaza-8-aza-guanosine;7-Methyl-8-oxo-guanosine;N2,N2-Dimethyl-6-thio-guanosine;N2-Methyl-6-thio-guanosine;1-Me-Guanosine Triphosphate;2'-Fluoro-N2-isobutyl-guanosine Triphosphate;2'-O-Methyl-N2-isobutyl-guanosine Triphosphate;2'-α-Ethynylguanosine Triphosphate;2'-a-Trifluoromethylguanosine triphosphate;2'-b-Ethynylguanosine triphosphate;2'-b-Trifluoromethylguanosine triphosphate;2'-Deoxy-2',2'-difluoroguanosine triphosphate;2'-Deoxy-2'-a-mercaptoguanosine triphosphate;2'-Deoxy-2'-a-thiomethoxyguanosine triphosphate;2'-Deoxy-2'-b-aminoguanosine triphosphate;2'-Deoxy-2'-b-azidoguanosine triphosphate;2'-Deoxy-2'-b-bromoguanosine triphosphate;2'-Deoxy-2'-b-chloroguanosine triphosphate;

[0077] 2'-Deoxy-2'-b-fluoroguanosine triphosphate;2'-Deoxy-2'-b-iodoguanosine triphosphate;2'-Deoxy-2'-b-mercaptoguanosine triphosphate;2'-Deoxy-2'-b-thiomethoxyguanosine triphosphate;4'-Azidoguanosine triphosphate;4'-Carbocyclic guanosine triphosphate;4'-Ethynylguanosine triphosphate;5'-Homoguanosine triphosphate;8-Bromoguanosine triphosphate;9-Deazaguanosine triphosphate;N2-Isobutyl-guanosine Triphosphate;1-Methylinosine;Inosine;1,2'-O-Dimethylinosine;7-Methylinosine;2'-O-Methylinosine;Epoxyqueuosine;Galactosylqueuosine;Mannosylqueuosine;Queuosine;Araiaminothymidine;Azathymidine;Deazathymidine;Deoxythymidine;2-Thiouridine;3-Methyluridine;5-Carboxymethyluridine;5-Hydroxyuridine;5-Methyluridine;5-Taurinomethyl-2-thiouridine 5-Taurinomethyluridine;Dihydrouridine;Pseudouridine;1-Methyl-3-(3-amino-5-carboxypropyl)pseudouridine;1-Methylpseudouridine;1-Ethylpseudouridine;2'-O-Methyluridine;2'-O-Methylpseudouridine;2'-O-Methyluridine;2'-O-Methyluridine;2-Thio-2'-O-methyluridine;3-(3-amino-3-carboxypropyl)uridine;3,2'-O-Dimethyluridine;3-Methylpseudouridine Triphosphate;4-Thiouridine;5-(Carboxyhydroxymethyl)uridine;5-(Carboxyhydroxymethyl)uridine methyl ester;5,2'-O-Dimethyluridine;5,6-Dihydro-uridine;5-Aminomethyl-2-thiouridine;5-Carbamoylmethyl-2'-O-methyluridine;5-Carbamoylmethyluridine;5-Carboxyhydroxymethyluridine;5-Carboxyhydroxymethyluridine methyl ester;5-Carboxymethylaminomethyl-2'-O-methyluridine;5-Carboxymethylaminomethyl-2-thiouridine;5-Carboxymethylaminomethyluridine;5-Carbamoylmethyluridine triphosphate;5-Methoxycarbonylmethyl-2'-O-methyluridine;5-Methoxycarbonylmethyl-2-thiouridine;5-Methoxycarbonylmethyluridine;5-Methoxyuridine;5-Methyl-2-thiouridine;5-Methylaminomethyl-2-selenouridine;5-Methylaminomethyl-2-thiouridine;5-Methylaminomethyluridine;5-Methyldihydrouridine;5-Oxyacetic acid-uridine triphosphate;5-Oxyacetic acid methyl ester-uridine triphosphate;N1-Methylpseudouracil;N1-Ethylpseudouracil;Uridine 5-oxyacetic acid;Uridine 5-oxyacetic acid methyl ester;3-(3-Amino-3-carboxypropyl)-uridine triphosphate;5-(Isopentenylaminomethyl)-2-thiouridine triphosphate;5-(Isopentenylaminomethyl)-2'-O-methyluridine Triphosphate;5-(Isopentenylaminomethyl)uridine triphosphate;5-Propynyluracil;α-Thio-uridine;1-(Aminoalkylaminocarbonylethylenyl)-2-(thio)pseudouracil;1-(Aminoalkylaminocarbonylethylenyl)-2,4-(dithio)pseudouracil;1-(Aminoalkylaminocarbonylethylenyl)-4-(thio)pseudouracil;1-(Aminoalkylaminocarbonylethylenyl)pseudouracil;1-(Aminocarbonylethylenyl)-2-(thio)pseudouracil 1-(aminocarbonylethylenyl)-2,4-(dithio)pseudouracil;1-(aminocarbonylethylenyl)-4-(thio)pseudouracil;1-(aminocarbonylethylenyl)pseudouracil;1-Substituted-2-(thio)pseudouracil;1-Substituted-2,4-(dithio)pseudouracil;1-Substituted-4-(thio)pseudouracil;1-Substituted pseudouracil;1-(aminoalkylamino-carbonylethylenyl)-2-(thio)-pseudouracil;1-Methyl-3-(3-amino-3-carboxypropyl)pseudouridine triphosphate;1-Methyl-3-(3-amino-3-carboxypropyl)pseudouridine triphosphate;1-Methyl-pseudouridine triphosphate1-Ethyl-pseudouridine triphosphate;2-(thio)pseudouracil;2'-Deoxyuridine;2'-Fluorouridine;2-(thio)uracil;2,4-(dithio)pseudouracil;2'-Methyl, 2'-amino, 2'-azido, 2'-fluoroguanosine;2'-Amino-2'-deoxyuridine triphosphate;2'-Azido-2'-deoxyuridine triphosphate;2'-Azidodeoxyuridine triphosphate;2'-Deoxyuridine;2'-Fluorouridine;2'-Deoxy-2'-α-aminouridine triphosphate;2'-Deoxy-2'-α-azidouridine Triphosphate;2-Methylpseudouridine;3-(3-amino-3-carboxypropyl)uracil;4-(Thio)pseudouracil;4-Thiouracil;5-(1,3-Diazol-1-alkyl)uracil;5-(2-Aminopropyl)uracil;5-(Aminoalkyl)uracil;5-(Dimethylaminoalkyl)uracil;5-(Guanidinium alkyl)uracil;5-(Methoxycarbonyl)uracil 5-(Carbonylmethyl)-2-(thio)uracil;5-(Methoxycarbonyl-methyl)uracil;5-(Methyl)-2-(thio)uracil;5-(Methyl)-2,4-(dithio)uracil;5-(Methyl)-4-(thio)uracil;5-(Methylaminomethyl)-2-(thio)uracil;5-(Methylaminomethyl)-2,4-(dithio)uracil;5-(Methylaminomethyl)-4-(thio)uracil 5-(Propynyl)uracil;5-(Trifluoromethyl)uracil;5-(2-Aminopropyl)uracil;5-(Alkyl)-2-(thio)pseudouracil;5-(Alkyl)-2,4-(dithio)pseudouracil;5-(Alkyl)-4-(thio)pseudouracil;5-(Alkyl)pseudouracil;5-(Alkyl)uracil;5-(Alkynyl)uracil;5-(Allylamino)uracil uracil;5-(cyanoalkyl)uracil;5-(dialkylaminoalkyl)uracil;5-(dimethylaminoalkyl)uracil;5-(guanidiniumalkyl)uracil;5-(halo)uracil;5-(1,3-diazol-1-alkyl)uracil;5-(methoxy)uracil;5-(methoxycarbonylmethyl)-2-(thio)uracil;5-(methoxycarbonylmethyl)uracil;

[0078] 5-(Methyl)-2-(thio)uracil;5-(Methyl)-2,4-(dithio)uracil;5-(Methyl)-4-(thio)uracil;5-(Methyl)-2-(thio)pseudouracil;5-(Methyl)-2,4-(dithio)pseudouracil;5-(Methyl)-4-(thio)pseudouracil;5-(Methyl)pseudouracil;5-(Methylaminomethyl)-2-(thio)uracil;5-(Methylaminomethyl)-2,4-(dithio)uracil )Uracil;5-(Methylaminomethyl)-4-(thio)uracil;5-(Propynyl)uracil;5-(Trifluoromethyl)uracil;5-Aminoallyl-uridine;5-Bromo-uridine;5-Iodo-uridine;5-Uracil;6-(Azo)uracil;6-Aza-uridine;Araiamino-uracil;Azauracil;Deazauracil;N3(Methyl)uracil;Pseudouridine triphosphate-1-2-ethanoic acid;Pseudouracil;4-Thio-pseudouridine Triphosphate;1-Carboxymethyl-pseudouridine;1-Methyl-1-deazapseudouridine;1-Propynyluridine;1-Taurinomethyl-1-methyluridine;1-Taurinomethyl-4-thiouridine;1-Taurinomethyl-pseudouridine;2-Methoxy-4-thiopseudouridine;2-Thio-1-methyl-1-deazapseudouridine;2-Thio-1-methylpseudouridine;2-Thio-5-azauridine;2-Thio-dihydropseudouridine;2-Thio-dihydrouridine;2-Thiopseudouridine;4-Methoxy-2-thiopseudouridine;4-Methoxypseudouridine;4-Thio-1-methylpseudouridine;4-Thiopseudouridine;5-Azauridine;Dihydropseudouridine;(±)1-(2-Hydroxypropyl)pseudouridine Triphosphate;(2R)-1-(2-hydroxypropyl)pseudouridine triphosphate;(2S)-1-(2-hydroxypropyl)pseudouridine triphosphate;(E)-5-(2-bromo-vinyl)aruridine triphosphate;(E)-5-(2-bromo-vinyl)uridine triphosphate;(Z)-5-(2-bromo-vinyl)aruridine triphosphate;(Z)-5-(2-bromo-vinyl)uridine triphosphate;1-(2,2,2-Trifluoroethyl)pseudouridine triphosphate;1-(2,2,3,3,3-Pentafluoropropyl)pseudouridine triphosphate;1-(2,2-Diethoxyethyl)pseudouridine triphosphate;1-(2,4,6-Trimethylbenzyl)pseudouridine triphosphate;1-(2,4,6-Trimethylbenzyl)pseudouridine triphosphate;1-(2,4,6-Trimethylphenyl)pseudouridine triphosphate;1-(2-Amino-2-carboxyethyl)pseudouridine triphosphate;1-(2-Aminoethyl)pseudouridine triphosphate;1-(2-Hydroxyethyl)pseudouridine triphosphate;1-(2-Methoxyethyl)pseudouridine triphosphate;1-(3,4-Bis-trifluoromethoxybenzyl)pseudouridine Triphosphate;1-(3,4-Dimethoxybenzyl)pseudouridine triphosphate;1-(3-Amino-3-carboxypropyl)pseudouridine triphosphate;1-(3-Aminopropyl)pseudouridine triphosphate;1-(3-Cyclopropyl-2-ynyl)pseudouridine triphosphate;1-(4-Amino-4-carboxybutyl)pseudouridine triphosphate;1-(4-Aminobenzyl)pseudouridine triphosphate;1-(4-Amino-butyl)pseudouridine triphosphate;1-(4-Amino-phenyl)pseudouridine triphosphate;1-(4-Azidobenzyl)pseudouridine triphosphate;1-(4-Bromobenzyl)pseudouridine triphosphate;1-(4-Chlorobenzyl)pseudouridine triphosphate;1-(4-Fluorobenzyl)pseudouridine Triphosphate;1-(4-iodobenzyl)pseudouridine triphosphate;1-(4-methanesulfonylbenzyl)pseudouridine triphosphate;1-(4-methoxybenzyl)pseudouridine triphosphate;1-(4-methoxybenzyl)pseudouridine triphosphate;1-(4-methoxy-phenyl)pseudouridine triphosphate;1-(4-methylbenzyl)pseudouridine triphosphate;1-(4-Nitrobenzyl)pseudouridine triphosphate;1-(4-Nitrophenyl)pseudouridine triphosphate;1-(4-Thiomethoxybenzyl)pseudouridine triphosphate;1-(4-Trifluoromethoxybenzyl)pseudouridine triphosphate;1-(4-Trifluoromethylbenzyl)pseudouridine triphosphate;1-(5-Aminopentyl)pseudouridine triphosphate;1-(6-Aminohexyl)pseudouridine triphosphate;1,6-Dimethyl-pseudouridine triphosphate;1-[3-(2-{2-[2-(2-aminoethoxy)ethoxy]-ethoxy}ethoxy)propionyl]pseudouridine triphosphate;1-{3-[2-(2-aminoethoxy)ethoxy]propionyl}pseudouridine Triphosphate;1-Acetylpseudouridine triphosphate;1-Alkyl-6-(1-propynyl)pseudouridine triphosphate;1-Alkyl-6-(2-propynyl)pseudouridine triphosphate;1-Alkyl-6-allylpseudouridine triphosphate;1-Alkyl-6-ethynylpseudouridine triphosphate;1-Alkyl-6-homoallylpseudouridine triphosphate

[0079] 1-Alkyl-6-vinylpseudouridine triphosphate;1-Allylpseudouridine triphosphate;1-Aminomethylpseudouridine triphosphate;1-Benzylpseudouridine triphosphate;1-Benzyl-pseudouridine triphosphate;1-Biotinyl-PEG2-pseudouridine triphosphate;1-Biotinylpseudouridine triphosphate;1-Butyl-pseudouridine triphosphate;1-Cyanomethylpseudouridine triphosphate;1-Cyclobutylmethyl-pseudouridine triphosphate;1-Cyclobutyl-pseudouridine triphosphate;1-Cycloheptylmethyl-pseudouridine triphosphate;1-Cyclohexylmethyl-pseudouridine Triphosphate;1-Cyclohexyl-pseudouridine triphosphate;1-Cyclooctylmethyl-pseudouridine triphosphate;1-Cyclooctyl-pseudouridine triphosphate;1-Cyclopentylmethyl-pseudouridine triphosphate;1-Cyclopentyl-pseudouridine triphosphate;1-Cyclopropylmethyl-pseudouridine triphosphate;1-Cyclopropyl-pseudouridine triphosphate;1-Hexyl-pseudouridine triphosphate;1-Homoallylpseudouridine triphosphate;1-Hydroxymethylpseudouridine triphosphate;1-Isopropyl-pseudouridine triphosphate;1-Me-2-thio-pseudouridine triphosphate;1-Me-4-thio-pseudouridine triphosphate;1-Me-alpha-thio-pseudouridine Triphosphate;1-Methanesulfonylmethylpseudouridine triphosphate;1-Methoxymethylpseudouridine triphosphate;1-Methyl-6-(2,2,2-trifluoroethyl)pseudouridine triphosphate;1-Methyl-6-(4-morpholino)-pseudouridine triphosphate;1-Methyl-6-(4-thiomorpholino)-pseudouridine triphosphate;1-Methyl-6-(substituted phenyl)pseudouridine triphosphate;1-Methyl-6-amino-pseudouridine triphosphate;1-Methyl-6-azido-pseudouridine triphosphate;1-Methyl-6-bromo-pseudouridine triphosphate;1-Methyl-6-butyl-pseudouridine triphosphate;1-Methyl-6-chloro-pseudouridine triphosphate;1-Methyl-6-cyano-pseudouridine triphosphate;1-Methyl-6-dimethylamino-pseudouridine triphosphate;1-Methyl-6-ethoxy-pseudouridine triphosphate;1-Methyl-6-ethylcarboxylate-pseudouridine triphosphate;1-Methyl-6-ethyl-pseudouridine triphosphate;1-Methyl-6-fluoro-pseudouridine triphosphate;1-Methyl-6-formyl-pseudouridine Triphosphate;1-Methyl-6-hydroxyamino-pseudouridine triphosphate;1-Methyl-6-hydroxy-pseudouridine triphosphate;1-Methyl-6-iodo-pseudouridine triphosphate;1-Methyl-6-isopropyl-pseudouridine triphosphate;1-Methyl-6-methoxy-pseudouridine triphosphate;1-Methyl-6-methylamino-pseudouridine triphosphate;1-Methyl-6-phenyl-pseudouridine triphosphate;1-Methyl-6-propyl-pseudouridine triphosphate;1-Methyl-6-tert-butyl-pseudouridine triphosphate;1-Methyl-6-trifluoromethoxy-pseudouridine triphosphate;1-Methyl-6-trifluoromethyl-pseudouridine triphosphate;1-Morpholinomethyl-pseudouridine triphosphate;1-Pentyl-pseudouridine Triphosphate;1-Phenyl-pseudouridine triphosphate;1-Pivaloylpseudouridine triphosphate;1-Propargylpseudouridine triphosphate;1-Propyl-pseudouridine triphosphate;1-Propynyl-pseudouridine;1-p-Tolyl-pseudouridine triphosphate;1-tert-Butyl-pseudouridine triphosphate;1-Thiomethoxymethylpseudouridine triphosphate;1-Thiomorpholinomethylpseudouridine triphosphate;1-Trifluoroacetylpseudouridine triphosphate;1-Trifluoromethylpseudouridine triphosphate;1-Vinylpseudouridine triphosphate;2,2'-Anhydro-uridine triphosphate;2'-Bromodeoxyuridine triphosphate;2'-F-5-Methyl-2'-deoxy-uridine triphosphate;2'-Methoxy-5-methyl-uridine triphosphate;2'-Methoxypseudouridine triphosphate;2'-α-Ethynyluridine triphosphate;2'-α-Trifluoromethyluridine triphosphate;2'-β-Ethynyluridine triphosphate;2'-β-Trifluoromethyluridine Triphosphate;2'-Deoxy-2',2'-difluorouridine triphosphate;2'-Deoxy-2'-a-mercaptouridine triphosphate;2'-Deoxy-2'-a-thiomethoxyuridine triphosphate;2'-Deoxy-2'-b-aminouridine triphosphate;2'-Deoxy-2'-b-azidouridine triphosphate;2'-Deoxy-2'-b-bromouridine triphosphate;2'-Deoxy-2'-b-chlorouridine triphosphate;

[0080] 2'-Deoxy-2'-b-fluorouridine triphosphate;2'-Deoxy-2'-b-iodouridine triphosphate;2'-Deoxy-2'-b-mercaptouridine triphosphate;2'-Deoxy-2'-b-thiomethoxyuridine triphosphate;2-Methoxy-4-thiouridine;2-Methoxyuridine;2'-O-Methyl-5-(1-propynyl)uridine triphosphate;3-Alkyl-pseudouridine triphosphate;4'-Azidouridine triphosphate;4'-Carbocyclic uridine triphosphate;4'-Ethynyluridine triphosphate;5-(1-Propynyl)-aruridine triphosphate;5-(2-Ruranyl)uridine triphosphate;5-Cyanouridine triphosphate;5-Dimethylaminouridine Triphosphate;5'-Homo-uridine triphosphate;5-Iodo-2'-fluoro-deoxyuridine triphosphate;5-Phenylethynyluridine triphosphate;5-Tritiomethyl-6-deuteriouridine triphosphate;5-Trifluoromethyl-uridine triphosphate;5-Vinylaruridine triphosphate;6-(2,2,2-Trifluoroethyl)pseudouridine triphosphate;6-(4-Morpholino)pseudouridine triphosphate;6-(4-Thiomorpholino)pseudouridine triphosphate;6-(Substituted phenyl)pseudouridine triphosphate;6-Aminopseudouridine triphosphate;6-Azidopseudouridine triphosphate;6-Bromopseudouridine triphosphate;6-Butylpseudouridine triphosphate;6-Chloropseudouridine Triphosphate;6-Cyanopseudouridine triphosphate;6-Dimethylaminopseudouridine triphosphate;6-Ethoxypseudouridine triphosphate;6-Ethylcarboxylate-pseudouridine triphosphate;6-Ethylpseudouridine triphosphate;6-Fluoropseudouridine triphosphate;6-Formylpseudouridine triphosphate;6-Hydroxyamino-pseudouridine triphosphate;6-Hydroxypseudouridine triphosphate;6-Iodopseudouridine triphosphate;6-Isopropylpseudouridine triphosphate;6-Methoxypseudouridine triphosphate;6-Methylaminopseudouridine triphosphate;6-Methylpseudouridine triphosphate;6-Phenylpseudouridine triphosphate;6-Propylpseudouridine triphosphate;6-tert-Butylpseudouridine triphosphate;6-Trifluoromethoxypseudouridine triphosphate;6-Trifluoromethylpseudouridine triphosphate;alpha-Thiopseudouridine triphosphate;Pseudouridine 1-(4-methylbenzenesulfonic acid)triphosphate;Pseudouridine 1-(4-methylbenzoic acid)triphosphate;Pseudouridine triphosphate 1-[3-(2-ethoxy)]propionic acid;Pseudouridine triphosphate 1-[3-{2-(2-[2-(2-ethoxy)ethoxy]ethoxy)ethoxy}]propionic acid;Pseudouridine triphosphate 1-[3-{2-(2-[2-{2(2-ethoxy)-ethoxy}-ethoxy]ethoxy)ethoxy}]propionic acid;Pseudouridine triphosphate 1-[3-{2-(2-[2-{2(2-ethoxy)-ethoxy}-ethoxy]ethoxy)ethoxy}]propionic acid;

[0081] Pseudouridine triphosphate 1-[3-{2-(2-ethoxy)-ethoxy}]propionic acid;Pseudouridine triphosphate 1-methylphosphonic acid;Pseudouridine triphosphate 1-methylphosphonic acid diethyl ester;Pseudouridine triphosphate-N1-3-propionic acid;Pseudouridine triphosphate-N1-4-butanoic acid;Pseudouridine triphosphate-N1-5-pentanoic acid;Pseudouridine triphosphate-N1-6-hexanoic acid;Pseudouridine triphosphate-N1-7-heptanoic acid;Pseudouridine triphosphate-N1-methyl-p-benzoic acid Triphosphate-N1-p-benzoic acid; Wybutosin; Hydroxywybutosin; Isowyosin; Peroxywybutosin; Undermodified hydroxywybutosin; 4-Demethylwybutosin; 2,6-(Diamino)purine; 1-(Aza)-2-(thio)-3-(aza)-phenoxazin-1-yl; 1,3-(diaza)-2-(oxo)-phenthiazin-1-yl; 1,3-(di Aza)-2-(oxo)-phenoxazin-1-yl; 1,3,5-(triaza)-2,6-(dioxa)naphthalene; 2(amino)purine; 2,4,5-(trimethyl)phenyl; 2'-Methyl, 2'-amino, 2'-azido, 2'-fluorocytidine; 2'-Methyl, 2'-amino, 2'-azido, 2'-fluoroadenine; 2'-Methyl, 2'-amino, 2'-azido, 2'-fluoroadenine Orolysine;2'-Amino-2'-deoxyribose;2-Amino-6-chloropurine;2-Azainosinyl;2'-Azido-2'-deoxyribose;2'-Fluoro-2'-deoxyribose;2'-Fluoro-modified bases;2'-O-methylribose;2-Oxo-7-aminopyridopyrimidin-3-yl;2-Oxo-pyridopyrimidin-3-yl;2-Pyridinone;3-Nitropyrrole;3-(Methyl)-7-(propynyl)isocarbostyrilyl;3-(Methyl)isocarbostyrilyl;4-(Fluoro)-6-(methyl)benzimidazole;4-(Methyl)benzimidazole;4-(Methyl)indolyl;4,6-(Dimethyl)indolyl;5-Nitroindole;5-Substituted pyrimidines;5-(Methyl)isocarbostyrilyl;5-Nitroindole6-(Aza)pyrimidine;6-(Azo)thymine;6-(Methyl)-7-(aza)indolyl;6-Chloropurine;6-Phenyl-pyrrolo-pyrimidin-2-one-3-yl;7-(Aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl;7-(Aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-(Aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(Aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(Aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl 7-(Guanidinium alkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-(Guanidinium alkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl;7-(Guanidinium alkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-(Guanidinium alkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-(Guanidinium alkylhydroxy)-1,3-(diaza)-2-(oxo-phenoxazin-1 -yl;7-(Guanidinium alkyl-hydroxy)-1,3-(diaza)-2-(oxo)-phenthiazin-1-yl;7-(Guanidinium alkyl-hydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl;7-(Propynyl)isocarbostyrilyl;7-(Propynyl)isocarbostyrilyl, 7-deazainosinyl;7-Substituted-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl;7-Substituted-1,3-(diaza)-2-(oxo) -Phenoxazin-1-yl;9-(methyl)-imidizopyridinyl;Aminoindolyl;Anthracenyl;Bis-ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl;Bis-ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl;Difluorotolyl;Hypoxanthine;Imidizopyridinyl;Inosinyl;Isocarbostyrilyl;Isoguanisine;N2-Substituted Purines;N6-Methyl-2-aminopurine;N6-Substituted Purines;N-Alkylated derivatives;Naphthalenyl;Nitrobenzimidazolyl;Nitroimidazolyl;Nitroindazolyl;Nitropyrazolyl;Nubularine;O6-Substituted purines;O-Alkylated derivatives;ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl;ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl;Oxoformycin Triphosphate;para-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl;para-substituted-6-phenyl-pyrrolo-pyrimidin-2-one-3-yl;Pentacenyl;Phenanthracenyl;Phenyl;Pyrenyl;Pyridopyrimidin-3-yl;2-Oxo-7-amino-pyridopyrimidin-3-yl;Pyrrolo-pyrimidin-2-one-3-yl;Pyrrolopyrimidinyl;Pyrrolopyridinyl;Stilbenzyl;Substituted 1,2,4-triazoles;Tetracenyl;Tubertidine;Xanthine;Xanthosine-5'-triphosphate;2-Thio-Zebularine;5-Aza-2-thiozebularine;7-Deaza-2-aminopurine;Pyridin-4-one ribonucleosides;2-Amino-riboside-triphosphate;Formycin A triphosphate; formycin B triphosphate; pyrrolosine triphosphate; 2'-hydroxyl-araadenosine triphosphate; 2'-hydroxyl-aracytidine triphosphate; 2'-hydroxyl-arauidine triphosphate; 2'-hydroxyl-araguanosine triphosphate; 5-(2-carbomethoxyvinyl)uridine triphosphate; and N6-(19-amino-pentaoxanonadecyl)adenosine triphosphate.

[0082] In some embodiments, the nucleobase or modified nucleobase of the nucleoside comprises a protecting group. Suitable protecting groups are described above. Those skilled in the art will understand that the selection of a protecting group depends on the nature of the nucleobase or modified nucleobase. For example, amines can be protected by Ac, iBu, Bn, or Bz.

[0083] Method for producing P-chiral phosphines The present disclosure provides a method for producing P-chiral phosphines. The synthetic method is shown in Scheme 1 below. Scheme 1 [ka]

[0084] Scheme 1 illustrates the modular phosphine synthesis enabled by the TLO-Psi reagent (1), which is based on an assembly process: loading step (R 1 ), coupling step (to introduce R 2 a substitution step (to introduce a leaving group followed by R 3 (introduction of P) and reduction steps. Net conformational retention of P is observed in all steps of this sequence, as confirmed by X-ray crystallography.

[0085] For clarity, compounds derived from either (-)- or (+)-TLO-Psi are designated with the suffixes a and b [a for (-) and b for (+)]. However, when only one reagent was available, simply changing the order of addition in this modular method could lead to either enantiomer of the coupling or displacement product (and vice versa).

[0086] In the process for producing chiral phosphines, it has been unexpectedly found that the P(V) reagent derived from trans-limonene oxide ("TLO-Psi") has superior reactivity to the P(V) reagent derived from cis-limonene oxide ("CLO-Psi"). Scheme 2 shows that the TLO-Psi-derived reagent 8a smoothly reacts with carbanion reagents to produce the thiophosphinic acid 9a and then the thiophosphinic ester 11a, whereas the CLO-Psi-derived reagent 3 does not. Scheme 2 [ka]

[0087] In one aspect, the present disclosure provides a method for producing a thiophosphinic acid, comprising: The carbanion reagent is a compound of formula (Ia): [ka] [In the ceremony R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl, where R 1 optionally, one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] with a compound of the formula:

[0088] In one embodiment, the thiophosphinic acid formed in the above method has the formula (II): [ka] [In the ceremony R 1 and R 2 are each independently C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl, where R 1 and R 2 each optionally containing one or more of the same or different R amay be substituted with a group; and R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl or C-linked heteroaryl] It is shown as follows.

[0089] In one embodiment, the carbanion reagent is an organolithium reagent R 2 Li, where R 2 is as defined above.

[0090] In one embodiment, the thiophosphinic acid formed in the above process has the formula (IIa): [ka] [In the formula, R 1 and R 2 is as defined above] or an enantiomer thereof.

[0091] In one embodiment, the process for producing thiophosphinic acids is carried out in an organic solvent, hi one embodiment, the organic solvent includes, but is not limited to, ether, THF, hexane, and dioxane.

[0092] In one embodiment, the method for preparing a thiophosphinic acid is carried out at room temperature for about 2 to about 12 hours, about 2 to about 10 hours, about 2 to about 8 hours, or about 2 to about 6 hours.

[0093] In one embodiment, the method further comprises reacting a thiophosphinic acid with an alkylating reagent to form a thiophosphinic ester.

[0094] In one embodiment, the thiophosphinate ester has the formula (III): [ka] [In the ceremony R 1 and R 2 are each independently C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R 1 and R 2 each optionally containing one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; and R' is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 alkynyl] It is expressed as:

[0095] In one embodiment, the thiophosphinate ester has the formula (IIIa): [ka] [In the formula: R 1 and R 2 is as defined above] or an enantiomer thereof.

[0096] In one embodiment, the thiophosphinate ester is [ka] is selected from the group consisting of:

[0097] In one embodiment, alkylating reagents include, but are not limited to, methyl iodide, ethyl iodide, and propyl iodide.

[0098] In one embodiment, the method for preparing a thiophosphinic ester is carried out in an organic solvent in the presence of a base at room temperature. In one embodiment, the organic solvent includes, but is not limited to, ether, THF, hexane, and dioxane. In one embodiment, the organic solvent is THF. In one embodiment, the base includes, but is not limited to, triethylamine, DIPEA, pyridine, 2,6-lutidine, and imidazole.

[0099] In one embodiment, the method further comprises reacting the thiophosphinate ester with an alkoxide agent to form an organophosphinate.

[0100] In one embodiment, the organic phosphinate has the formula (IV): [ka] [In formula: R 1 and R 2 are each independently C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R 1 and R 2 each optionally containing one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10aryl, or C-linked heteroaryl; and R' is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 alkynyl] It is shown as follows.

[0101] In one embodiment, the organic phosphinate has the formula (IVa): [ka] or an enantiomer thereof.

[0102] In one embodiment, the organic phosphinate is [ka] is selected from the group consisting of:

[0103] In one embodiment, the method for producing phosphine oxide is carried out in an organic solvent. In one embodiment, the organic solvent includes, but is not limited to, alcohol, ether, THF, hexane, and dioxane. In one embodiment, the method is carried out in an alcohol. In one embodiment, the method is carried out at room temperature for about 4 hours.

[0104] In one embodiment, the method further comprises reacting the organic phosphinate with a carbanion reagent to form a phosphine oxide.

[0105] In one embodiment, the phosphine oxide has the formula (V): [ka] [In the ceremony R 1 , R 2 , and R 3 are each independently C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R 1 , R 2 , and R 3 each optionally containing one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] It is shown as follows.

[0106] In one embodiment, the phosphine oxide has the formula (Va): [ka] or an enantiomer thereof.

[0107] In one embodiment, the carbanion reagent is R 3 MgBr is a Grignard reagent, where R 3 is as defined above.

[0108] In one embodiment, the Grignard reagent is [ka] is selected from the group consisting of:

[0109] In one embodiment, the phosphine oxide is [ka] is selected from the group consisting of:

[0110] Enantiomerically pure [R 1 R 2When preparing thiophosphinic acids represented by [P(O)SH], a search for the optimal leaving group during the final substitution reaction is conducted. Table 5 below shows that chlorophosphine sulfides (entry 1) are inactive toward nucleophilic attack, whereas chlorophosphine oxides (entry 2) react readily, but with a low enantiomeric ratio ("er"). In contrast, thiophosphinic esters (alkylation of the free SH) function as efficient leaving groups. Surprisingly, using methoxy as the leaving group increases the er ratio of the substitution reaction to 98:2 while maintaining high reactivity. [Table 8]

[0111] In one embodiment, the process for producing phosphine oxide is carried out in an organic solvent. In one embodiment, the organic solvent includes, but is not limited to, ether, THF, hexane, and dioxane. In one embodiment, the process is carried out in THF. In one embodiment, the process is carried out at -78°C for about 2 to about 12 hours, about 2 to about 10 hours, about 2 to about 8 hours, about 2 to about 6 hours, or about 2 to about 4 hours.

[0112] In one embodiment, the method further comprises reacting the phosphine oxide with a reducing agent to form a phosphine.

[0113] In one embodiment, the phosphine is of formula (VI): [ka] [In the ceremony R 1 , R 2 , and R 3 independently, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R1 , R 2 , and R 3 each optionally containing one or more of the same or different R a may be substituted with a group; and R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] It is a compound represented by the formula:

[0114] In one embodiment, the phosphine has the formula (VIa): [ka] or an enantiomer thereof.

[0115] In one embodiment, reducing agents include, but are not limited to, HSiCl3 (with or without Et3N), HSiCl3 with PPh3 as a sacrificial, polymethylhydrosiloxane (PMHS) and Ti(OiPr)4, and MeOTf / Meerwain salt, and NaBH4.

[0116] In one embodiment, the method for producing phosphines is carried out in an organic solvent. In one embodiment, the organic solvent includes, but is not limited to, dichloromethane, ether, THF, hexane, and dioxane. In one embodiment, the method is carried out in dichloromethane. In one embodiment, the method is carried out at 50° C. for about 2 to about 4 hours.

[0117] The above reaction conditions are exemplary and not intended to be limiting. Those skilled in the art will understand that reaction conditions such as reaction time and temperature, solvent identity and amount, etc., can be varied according to methods known in the art.

[0118] Methods for producing oligonucleotides The present disclosure provides methods for producing oligonucleotides, the synthesis of which is depicted in Scheme 3 below.

[0119] Scheme 3 shows an efficient workflow for generating dinucleotides with methyl phosphonate linkages. After the methyl is introduced into the TLO-Psi reagent, compounds 12a or 12b are reacted with a nucleoside to form (R p )-12 or (S p )-12, resulting in organophosphorus compounds loaded with nucleosides. Each compound is then coupled with a second nucleoside to yield dinucleotides with high stereospecificity. Scheme 3 [ka]

[0120] In the development of the above dinucleotides, it was unexpectedly found that the P(V) reagent derived from trans-limonene oxide ("TLO-Psi") has superior reactivity to the P(V) reagent derived from cis-limonene oxide ("CLO-Psi"). Scheme 4 shows that compound 12a readily reacts with 1-adamantyl methanol (Ad-CH2-OH) to give the (S p )-6, while compound 5 shows no such reactivity. Scheme 4 [ka]

[0121] In one aspect, the disclosure provides a method for preparing a nucleoside-loaded organophosphorus compound, comprising: coupling a nucleoside to a compound of formula (Ia): [ka] [In formula: R 1is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl, where R 1 optionally, one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] or a salt thereof, an enantiomer thereof, or a combination thereof to form a nucleoside-loaded organophosphorus compound.

[0122] In one embodiment, the nucleoside-loaded organophosphorus compound has the formula (VII) or (VIIa): [ka] [In the ceremony R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; R' is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; Nu 1 is a nucleoside, Here, Nu 1 may optionally contain, independently of one another, one or more of the same or different modifications or one or more of the same or different protecting groups. or a diastereomer thereof.

[0123] In one embodiment, R 1 and each of R' is -CH3.

[0124] In one embodiment, Nu 1 teeth, [ka] is a nucleoside selected from the group consisting of: where: T is [ka] and; A Bz teeth [ka] and; G iBu teeth [ka] and C Bz teeth [ka] is.

[0125] In one embodiment, the method for preparing the nucleoside-loaded organophosphorus compound is carried out in an organic solvent, including but not limited to ether, THF, hexane, and dioxane.

[0126] In one embodiment, the method for preparing a nucleoside-loaded organophosphorus compound is carried out at room temperature for about 2 to about 12 hours, about 2 to about 10 hours, about 2 to about 8 hours, or about 2 to about 6 hours.

[0127] In one embodiment, the method for preparing a nucleoside-loaded organophosphorus compound is carried out in the presence of a base, including but not limited to DBU, triethylamine, DIPEA, pyridine, 2,6-lutidine, and imidazole.

[0128] In one embodiment, the method further comprises reacting the nucleoside-loaded organophosphorus compound with a second nucleoside to form a dinucleotide.

[0129] In one embodiment, the dinucleotide has the formula (VIII) or (VIIIa): [ka] [In formula: R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; Nu 1 and Nu 2 each of which is a nucleoside; Here, Nu 1 and Nu 2 may optionally contain, independently of each other, one or more of the same or different modifications or one or more of the same or different protecting groups. It is a compound represented by the formula:

[0130] In one embodiment, R 1 is -CH3;Nu 1 and Nu 2 are each independently [ka] is a nucleoside selected from the group consisting of: where T is [ka] and; A Bz teeth [ka] and; G iBu teeth [ka] and C Bz teeth [ka] is.

[0131] In one embodiment, the method for producing the dinucleotide is carried out in an organic solvent. In one embodiment, the organic solvent includes, but is not limited to, ether, THF, hexane, and dioxane.

[0132] In one embodiment, the method for producing the dinucleotide is carried out at room temperature for about 2 to about 16 hours, about 2 to about 14 hours, about 2 to about 12 hours, or about 2 to about 10 hours.

[0133] In one embodiment, the method for producing the dinucleotide is carried out in the presence of a base, which in one embodiment includes, but is not limited to, DBU, triethylamine, DIPEA, pyridine, 2,6-lutidine, and imidazole.

[0134] In one embodiment, the method comprises: a) reacting the dinucleotide with a compound of formula (Ia) to form a loaded dinucleotide; b) reacting the loaded dinucleotide formed in step (a) with a third nucleoside to form a trinucleotide; c) repeating steps (a) and (b) one or more times to form an oligonucleotide having a desired number of nucleotides. It further includes:

[0135] In one embodiment, the reaction conditions for step a) of the method for producing an oligonucleotide are the same as the reaction conditions for the method for producing a nucleoside-loaded organophosphorus compound. In one embodiment, the reaction conditions for step b) of the method for producing an oligonucleotide are the same as the reaction conditions for the method for producing a dinucleotide.

[0136] In one embodiment, the dinucleotide has the formula (VIII) or (VIIIa): [ka] [In formula: R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, or C 2-6 is alkynyl; Nu 1 and Nu 2 each of which is a nucleoside; Here, Nu 1 and Nu 2 may optionally, independently of one another, include one or more of the same or different modifications or one or more of the same or different protecting groups. or an enantiomer thereof.

[0137] In one embodiment, the method of forming a dinucleotide comprises: a) Reconstituting a dinucleotide with formula C: [ka] or an enantiomer thereof to form a loaded chimeric dinucleotide; and b) reacting the loaded chimeric dinucleotide formed in step (a) with a third nucleoside to form a chimeric trinucleotide bearing phosphonate and phosphorothioate linkages. Further includes:

[0138] In one embodiment, step a) of the method of forming the chimeric trinucleotide is carried out in an organic solvent, including but not limited to ACN, ether, THF, hexane, and dioxane.

[0139] In one embodiment, the method for preparing a nucleoside-loaded organophosphorus compound is carried out at room temperature for about 2 to about 12 hours, about 2 to about 10 hours, about 2 to about 8 hours, or about 2 to about 6 hours.

[0140] In one embodiment, the method for preparing a nucleoside-loaded organophosphorus compound is carried out in the presence of a base, including but not limited to DBU, triethylamine, DIPEA, pyridine, 2,6-lutidine, and imidazole.

[0141] The above reaction conditions are exemplary and not intended to be limiting. Those skilled in the art will understand that reaction conditions such as reaction time and temperature, solvent identity and amount, etc., can be varied according to methods known in the art.

[0142] Methods for preparing compounds of formula (Ia) In one aspect, the present disclosure provides a compound of formula (Ia): [ka] [In formula: R 1 is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl; where R 1 optionally, one or more of the same or different R a may be substituted with a group; R a is deuterium, CD3, C 1-6 Alkyl, C 2-6 Alkenyl, C2-6 Alkynyl, halogen, CF3, C 1-6 Alkoxy, C 3-10 Cycloalkyl, C 6-10 aryl, or C-linked heteroaryl] or an enantiomer thereof, comprising: The carbanion reagent is reacted with a compound of Formula A: [ka] or an enantiomer thereof to form a compound of formula (Ia).

[0143] In one embodiment, the carbanion reagent is a Grignard reagent. In one embodiment, the Grignard reagent is R 1 MgBr, where R 1 is as defined above.

[0144] In one embodiment, the Grignard reagent is [ka] is selected from the group consisting of:

[0145] In one embodiment, the method for preparing a compound of formula (Ia) is carried out at room temperature for about 2 to about 12 hours, about 2 to about 10 hours, about 2 to about 8 hours, or about 2 to about 6 hours.

[0146] In one embodiment, the process for preparing the compound of formula (Ia) is carried out in an organic solvent. In one embodiment, the organic solvent includes, but is not limited to, ether, THF, hexane, and dioxane.

[0147] The above reaction conditions are exemplary and not intended to be limiting. Those skilled in the art will understand that reaction conditions such as reaction time and temperature, solvent identity and amount, etc., can be varied according to methods known in the art.

[0148] definition Unless otherwise stated, the following terms used in this application, including the specification and claims, have the definitions set forth below. It should be noted that as used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Conventional methods of mass spectroscopy, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are utilized unless otherwise indicated. In this application, the use of "or" or "and" means "and / or" unless otherwise indicated. Furthermore, the use of the word "comprising," as well as other forms such as "comprises" and "includes," is not limiting.

[0149] Units, prefixes, and symbols are written in the format recognized by the International System of Units (SI). Numerical ranges are inclusive of the numbers defining the range. When a range of numerical values ​​is disclosed, it is to be understood that each intervening integer and fractional value between the stated upper and lower limits of the range is also specifically disclosed, along with each subgroup therebetween. The upper and lower limits of any range can be independently included or excluded, and each range is also encompassed within the scope of the invention when either, neither, or both limits are included. When a numerical value is explicitly stated, it is to be understood that numerical values ​​that are approximately the same amount or value as the stated numerical value are also within the scope of the invention. When a combination is disclosed, each subcombination of the elements of that combination is also specifically disclosed and is within the scope of the invention. Conversely, when different elements or groups of elements are individually disclosed, the combination is also disclosed. Where any element of the invention is disclosed as having multiple alternatives, examples of the invention in which each alternative is excluded alone or in any combination with the other alternatives are also disclosed herein; multiple elements of the invention can include such exclusions, and all combinations of elements including such exclusions are disclosed herein.

[0150] The present disclosure is intended to encompass all isotopes of atoms present in the compounds of the present invention. Isotopes include those atoms having the same atomic number but different mass numbers. By way of general example and without limitation, isotopes of hydrogen include deuterium and tritium. Isotopes of hydrogen include 1 H (hydrogen), 2 H (deuterium) and 3 H (tritium). They are also commonly designated D for deuterium and T for tritium. In this application, CD3 refers to a methyl group in which all hydrogen atoms are deuterium. The carbon isotopes are 13 C and 14 C. Isotopically labeled compounds of the present invention may generally be prepared by techniques known to those skilled in the art, or by methods analogous to those described herein, substituting an appropriately isotopically labeled reagent for the unlabeled reagent otherwise utilized.

[0151] As used herein, the term "stereoisomer" refers to all of the different possible isomeric and conformational forms that a compound (e.g., a compound of any formula described herein) may possess, and in particular to all possible stereochemically and conformationally isomeric forms, all diastereomers, enantiomers, and / or conformers of the basic molecular structure. Some compounds of the present disclosure may exist in different tautomeric forms, all of which are encompassed within the scope of the present disclosure.

[0152] In this disclosure, the term "enantiomer" refers to each individual optically active form of a compound of the present disclosure.

[0153] In this disclosure, the term "diastereomers" refers to stereoisomers that are not mirror images of each other and are not superimposable with respect to one another.

[0154] In this disclosure, the term "nucleic acid" encompasses poly- or oligo-ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from nucleobases and / or modified nucleobase N- or C-glycosides; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate bridges and / or modified phosphorus atom bridges. The term encompasses nucleic acids containing any combination of nucleobases, modified nucleobases, sugars, modified sugars, phosphonate bridges, or modified phosphorus atom bridges. Examples include, but are not limited to, nucleic acids containing ribose moieties, nucleic acids containing deoxyribose moieties, nucleic acids containing ribose and deoxyribose moieties, and nucleic acids containing ribose and modified ribose moieties. The prefix "poly-" refers to a nucleic acid containing from about 1 to about 10,000 nucleotide monomer units, and the prefix "oligo-" refers to a nucleic acid containing from about 1 to about 200 nucleotide monomer units. The term "nucleic acid" may also encompass CDNs.

[0155] In this disclosure, the terms "nucleobase" and "nucleoside base moiety," used interchangeably, refer to the portion of a nucleic acid that participates in hydrogen bonding to link one nucleic acid strand to a complementary strand in a sequence-specific manner. The most common naturally occurring nucleobases are adenine (A), guanine (G), uracil (U), cytosine (C), and thymine (T).

[0156] In the present disclosure, nucleobases may be represented by the abbreviations A, G, U, C, T, and Hyp. The abbreviation A refers to adenine; G refers to guanine; U refers to uracil; C refers to cytosine; T refers to thymine; and Hyp refers to hypoxanthine.

[0157] In this disclosure, the terms "modified nucleobase" and "modified nucleoside base moiety," used interchangeably, refer to moieties that can replace nucleobases. Modified nucleobases mimic the spatial arrangement, electronic properties, or some other physicochemical properties of nucleobases and retain the hydrogen-bonding properties that bind one nucleic acid strand to another in a sequence-specific manner. Modified nucleobases can generally pair with naturally occurring bases (e.g., uracil, thymine, adenine, cytosine, guanine) without substantially affecting the melting behavior, intracellular enzyme recognition, or activity of oligonucleotide duplexes. The terms "modified nucleobase" and "modified nucleoside base moiety," used interchangeably, further encompass heterocyclic compounds that can function as nucleoside bases, including certain "universal bases" that are not nucleoside bases in the most classical sense but function as nucleoside bases. Of particular mention as a universal base is 3-nitropyrrole.

[0158] In this disclosure, the term "nucleoside" refers to a glycosylamine compound in which a nucleobase (a nitrogenous base such as adenine, guanine, thymine, uracil, 5-methyluracil, etc.) or modified nucleobase is covalently linked to a five-carbon sugar (ribose or deoxyribose) or modified sugar.

[0159] In the present disclosure, the term "sugar" refers to closed and / or open monosaccharides. Sugars include, but are not limited to, ribose, deoxyribose, pentofuranose, pentopyranose, morpholino, carbocyclic analogs, hexopyranose moieties, and bicyclic sugars such as those found in locked nucleic acids. Examples of locked nucleic acids include, but are not limited to, the nucleic acids disclosed in WO2016 / 079181.

[0160] In this disclosure, the term "modified sugar" refers to a moiety that can replace a sugar. The modified sugar mimics the spatial arrangement, electronic properties, or some other physicochemical property of the sugar.

[0161] In this disclosure, the term "nucleotide" refers to a moiety in which a nucleobase or modified nucleobase is covalently linked to a sugar or modified sugar, and the sugar or modified sugar is covalently linked to a modified phosphorus atom moiety, such as a phosphate or thiophosphate group.

[0162] In this disclosure, the term "peptide" refers to a chain of amino acid monomers linked by peptide bonds. Generally, peptides will have about 50 amino acids or less. The term "peptide" encompasses both naturally occurring and non-naturally occurring amino acids. Peptides can be linear or cyclic.

[0163] In this disclosure, the term "protein" includes one or more polypeptides arranged in a biologically functional manner. Examples of biologically functional proteins include, but are not limited to, enzymes, antibodies, cytokines, hormones, transmembrane proteins, etc.

[0164] In this disclosure, the term "moiety" refers to a specific segment or functional group of a molecule. A chemical moiety is often a recognized chemical entity that is embedded in or attached to a molecule.

[0165] In this disclosure, the terms "solid support" or "resin," used interchangeably herein, refer to any support that allows for the large-scale synthesis of nucleic acids and / or peptides and can be reused when necessary. As used herein, the terms refer to a polymer that is insoluble in the media used in the reaction steps performed to synthesize nucleic acids and / or peptides and that has been derivatized to contain reactive groups.

[0166] In this disclosure, the term "linking moiety" refers to any moiety that may be optionally positioned between the terminal nucleoside and the solid support, or between the terminal nucleoside and another nucleoside, nucleotide, or nucleic acid.

[0167] In this disclosure, the term "purified," when used in reference to nucleic acids, refers to something that is separated from at least one contaminant. As used herein, a "contaminant" is any substance that renders another substance, apart from the other substance, unsuitable, impure, or inferior. Thus, a purified oligonucleotide exists in a form or setting that is different from that in which it existed prior to being subjected to the purification method.

[0168] In this disclosure, the term "alkyl" as used by itself or as part of another group refers to an alkyl group having 1 to 12 carbon atoms (i.e., C 1-12 alkyl), or an unsubstituted straight or branched chain aliphatic hydrocarbon containing the specified number of carbon atoms, e.g., C alkyl such as methyl, C alkyl such as ethyl, C alkyl such as propyl or isopropyl, etc. In one embodiment, an alkyl group is C 1-10 In another embodiment, the alkyl group is C 1-6 In another embodiment, the alkyl group is C 1-4 A non-limiting example of C is alkyl. 1-10 Alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, iso-butyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, and decyl. 1-6 Alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, isobutyl, and hexyl. 1-4 Alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, and isobutyl.

[0169] In the present disclosure, the term "optionally substituted alkyl" refers to one or more R a is an alkyl group, as defined above, having a group.

[0170] In this disclosure, the term "cycloalkyl" as used by itself or as part of another group refers to a cycloalkyl group having from 3 to 12 carbon atoms (i.e., C 3-12 Cycloalkyl) refers to an unsubstituted saturated and partially unsaturated cyclic aliphatic hydrocarbon, e.g., containing one or two double bonds, containing one to three rings, having the specified number of carbon atoms. In one embodiment, a cycloalkyl group has two rings. In one embodiment, a cycloalkyl group has one ring. In another embodiment, a cycloalkyl group is saturated. In another embodiment, a cycloalkyl group is unsaturated. In another embodiment, a cycloalkyl group is C 3-8 In another embodiment, the cycloalkyl group is C 3-7 In another embodiment, the cycloalkyl group is C 5-7 In another embodiment, the cycloalkyl group is C 3-6 A cycloalkyl group. The term "cycloalkyl" includes groups in which the ring CH2- is replaced with -C(=O)-. Non-limiting exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decalin, adamantyl, cyclohexenyl, cyclopentenyl, cyclohexenyl, and cyclopentanone.

[0171] In the present disclosure, the term "optionally substituted cycloalkyl" refers to one or more R a is a cycloalkyl group as defined above having the group.

[0172] The term "optionally substituted cycloalkyl" includes cycloalkyl groups fused to an optionally substituted aryl, such as phenyl, or fused to an optionally substituted heteroaryl, such as pyridyl. An optionally substituted cycloalkyl having a fused optionally substituted aryl or fused optionally substituted heteroaryl group may be attached to the remainder of the molecule at any available carbon atom on the cycloalkyl ring. In one embodiment, the optionally substituted cycloalkyl group is a 5-, 6-, or 7-membered cycloalkyl group fused to a phenyl group, where the phenyl may be optionally substituted with one, two, or three substituents.

[0173] In this disclosure, the term "alkenyl," as used by itself or as part of another group, refers to an alkyl containing one, two, or three carbon-carbon double bonds. In one embodiment, an alkenyl group is C 2-6 In another embodiment, the alkenyl group is a C2-4 alkenyl group. Non-limiting exemplary alkenyl groups include ethenyl, propenyl, isopropenyl, butenyl, sec-butenyl, pentenyl, and hexenyl.

[0174] In this disclosure, the term "optionally substituted alkenyl" as used herein by itself or as part of another group refers to an alkenyl that is unsubstituted or has one or more R a It refers to an alkenyl which is either substituted or unsubstituted.

[0175] In this disclosure, the term "alkynyl," as used by itself or as part of another group, refers to an alkyl containing one to three carbon-carbon triple bonds. In one embodiment, the alkynyl has one carbon-carbon triple bond. In one embodiment, the alkynyl group is C 2-6In another embodiment, the alkynyl group is a C2-4 alkynyl group. Non-limiting exemplary alkynyl groups include ethynyl, propynyl, butynyl, 2-butynyl, pentynyl, and hexynyl.

[0176] In this disclosure, the term "optionally substituted alkynyl" as used herein by itself or as part of another group refers to an alkynyl group that is unsubstituted or has one or more R a It refers to an alkynyl which is either substituted or unsubstituted.

[0177] In this disclosure, the term "aryl," as used by itself or as part of another group, refers to a group containing an unsubstituted aromatic ring system. In one embodiment, an aryl group is a monocyclic, bicyclic, or polycyclic aromatic ring having from 5 to 14 carbon atoms, i.e., C 5-14 Aryl, C 6-12 Aryl, C 6-10 Aryl, or C 6-8 It has an aryl. Non-limiting exemplary aryl groups include phenyl (abbreviated as "Ph"), naphthyl, phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylenyl, and fluorenyl groups. In one embodiment, the aryl group is phenyl or naphthyl. In one embodiment, the aryl group includes an organometallic group in which the aromatic ring is bonded to a metal atom. Non-limiting exemplary aryl groups include ferrocene and cobaltocene. The group or point of attachment is on the aromatic ring.

[0178] In this disclosure, the term "optionally substituted aryl" as used herein by itself or as part of another group refers to an aryl that is unsubstituted or substituted with one or more R a It refers to an aryl which is either substituted or unsubstituted by a substituent.

[0179] In one embodiment, the optionally substituted aryl is an optionally substituted phenyl. In one embodiment, the optionally substituted phenyl has four substituents. In another embodiment, the optionally substituted phenyl has three substituents. In another embodiment, the optionally substituted phenyl has two substituents. In another embodiment, the optionally substituted phenyl has one substituent. Non-limiting exemplary substituted aryl groups include 2-methylphenyl, 2-methoxyphenyl, 2-fluorophenyl, 2-chlorophenyl, 2-bromophenyl, 3-methylphenyl, 3-methoxyphenyl, 3-fluorophenyl, 3-chlorophenyl, 4-methylphenyl, 4-ethylphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl, 2,6-difluorophenyl, 2,6-dichlorophenyl, 2-methyl, 3-methoxyphenyl, 2-ethyl, 3-methoxyphenyl, 3,4-dimethoxyphenyl, 3,5-difluorophenyl 3,5-dimethylphenyl, 3,5-dimethoxy, 4-methylphenyl, 2-fluoro-3-chlorophenyl, and 3-chloro-4-fluorophenyl. The term "optionally substituted aryl" encompasses phenyl groups fused to optionally substituted cycloalkyls and phenyl groups fused to optionally substituted heterocyclocycles. The optionally substituted cycloalkyl-fused and optionally substituted heterocycle-fused, optionally substituted aryl are bonded to the rest of the molecule at any available carbon atom on the aryl ring. [ka] Examples include:

[0180] In this disclosure, the term "aryloxy" as used by itself or as part of another group refers to an optionally substituted aryl attached to a terminal oxygen atom. A non-limiting exemplary aryloxy group is PhO-.

[0181] As used herein, the terms "heterocycle," "heterocyclyl," or "heterocyclic group" refer to a stable 3-, 4-, 5-, 6-, or 7-membered monocyclic or bicyclic, or 7-, 8-, 9-, 10-, 11-, 12-, 13-, or 14-membered polycyclic heterocyclic ring that is saturated, partially unsaturated, or fully unsaturated and contains carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, and S, and are intended to include any polycyclic group in which any of the above heterocyclic rings are fused to a benzene ring. The nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., N→O and S(O)). p (where p is 0, 1, or 2). The nitrogen atom can be substituted or unsubstituted (i.e., N or NR, where R, if specified, is H or another substituent). A heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. The heterocyclic rings described herein can be substituted on a carbon or nitrogen atom if the resulting compound is stable. A nitrogen in a heterocycle can optionally be quaternized. When the total number of S and O atoms in the heterocycle exceeds 1, then it is preferred that these heteroatoms are not adjacent to one another. It is preferred that the total number of S and O atoms in the heterocycle does not exceed 1. When the term "heterocycle" is used, it is intended to include heteroaryl.

[0182] Examples of heterocycles include, but are not limited to, acridinyl, azetidinyl, azotinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrazolyl, and 2H,6H-dithiazinyl. Hydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, imidazolopyridinyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isothiazolopyridinyl, isoxazolyl, isoxazolopyridinyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolopyridinyl, oxazolidinyl, perimidinyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, phteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, Pyrazolinyl, pyrazolopyridinyl, pyrazolyl, pyridazinyl, pyridooxazolyl, pyridoimidazolyl, pyridothiazolyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2-pyrrolidonyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrazolyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,Examples include 3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thiazolopyridinyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl. Also included are fused ring and spiro compounds containing, for example, the above heterocycles.

[0183] As used herein, the term "bicyclic heterocycle" or "bicyclic heterocyclic group" refers to a stable 9- or 10-membered heterocyclic ring system containing two fused rings and consisting of carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, and S. Of the two fused rings, one ring is a 5- or 6-membered monocyclic aromatic ring, including a 5-membered heteroaryl ring, a 6-membered heteroaryl ring, or a benzo ring, each fused to a second ring. The second ring is a saturated, partially unsaturated, or unsaturated 5- or 6-membered monocyclic ring, including a 5-membered heterocycle, a 6-membered heterocycle, or a carbocycle (provided that when the second ring is a carbocycle, the first ring is a ring other than benzo).

[0184] Bicyclic heterocyclic group can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure.Bicyclic heterocyclic group described herein can be substituted on carbon or nitrogen atom as long as the resulting compound is stable.If the total number of S and O atoms in the heterocycle exceeds 1, then it is preferred that these heteroatoms are not adjacent to each other.It is preferred that the total number of S and O atoms in the heterocycle is 1 or less.

[0185] Examples of bicyclic heterocyclic groups include, but are not limited to, quinolinyl, isoquinolinyl, phthalazinyl, quinazolinyl, indolyl, isoindolyl, indolinyl, 1H-indazolyl, benzimidazolyl, 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydroquinolinyl, 2,3-dihydrobenzofuranyl, chromanyl, 1,2,3,4-tetrahydroquinoxalinyl, and 1,2,3,4-tetrahydroquinazolinyl.

[0186] Bridged rings are also included in the definition of heterocycle. A bridged ring occurs when one or more, preferably one to three, atoms (i.e., C, O, N, or S) link two non-adjacent carbon or nitrogen atoms. Examples of bridged rings include, but are not limited to, one carbon atom, two carbon atoms, one nitrogen atom, two nitrogen atoms, and a carbon-nitrogen group. It is noted that a bridge always converts a monocyclic ring into a tricyclic ring. When a ring is bridged, the substituents recited for the ring may also be present on the bridge.

[0187] The term "heterocyclylalkyl" refers to a heterocyclyl or substituted heterocyclyl bonded to an alkyl group attached to the core of a compound.

[0188] As used herein, the term "aromatic heterocyclic group" or "heteroaryl" refers to stable monocyclic and polycyclic aromatic hydrocarbons containing at least one heteroatom ring member, such as sulfur, oxygen, or nitrogen. Heteroaryl groups include, but are not limited to, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, furyl, quinolyl, isoquinolyl, thienyl, imidazolyl, thiazolyl, indolyl, pyrroyl, oxazolyl, benzofuryl, benzothienyl, benzthiazolyl, isoxazolyl, pyrazolyl, triazolyl, tetrazolyl, indazolyl, 1,2,4-thiadiazolyl, isothiazolyl, purinyl, carbazolyl, benzimidazolyl, indolinyl, benzodioxolanyl, and benzodioxane. Heteroaryl groups may be substituted or unsubstituted. The nitrogen atoms may be substituted or unsubstituted (i.e., N or NR, where R, as defined, is H or another substituent). The nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., N→O and S(O)). p where p is 0, 1, or 2).

[0189] In one embodiment, the term "heteroaryl" or "heteroaromatic" refers to unsubstituted monocyclic and bicyclic aromatic ring systems having 5 to 14 ring atoms, i.e., 5- to 14-membered heteroaryls, in which at least one carbon atom in a ring is replaced with a heteroatom independently selected from the group consisting of oxygen, nitrogen, and sulfur. In one embodiment, heteroaryls contain 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of oxygen, nitrogen, and sulfur. In one embodiment, heteroaryls have 3 heteroatoms. In another embodiment, heteroaryls have 2 heteroatoms. In another embodiment, heteroaryls have 1 heteroatom. In another embodiment, heteroaryls are 5- to 10-membered heteroaryls. In another embodiment, heteroaryls are 5- or 6-membered heteroaryls. In another embodiment, heteroaryls have 5 ring atoms, e.g., thienyl, a 5-membered heteroaryl having 4 carbon atoms and 1 sulfur atom. In another embodiment, the heteroaryl has six ring atoms, e.g., pyridyl, a six-membered heteroaryl having five carbon atoms and one nitrogen atom. Non-limiting exemplary heteroaryl groups include thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl, furyl, benzofuryl, pyranyl, isobenzofuranyl, benzoxazolyl, chromenyl, xanthenyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, isoindolyl, 3H-indolyl, and indolyl. , indazolyl, purinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, cinnolinyl, quinazolinyl, phtheridinyl, 4aH-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, thiazolyl, isothiazolyl, phenothiazolyl, isoxazolyl, furazanyl, and phenoxazinyl.In one embodiment, heteroaryl is thienyl (e.g., thien-2-yl and thien-3-yl), furyl (e.g., 2-furyl and 3-furyl), pyrrolyl (e.g., 1H-pyrrol-2-yl and 1H-pyrrol-3-yl), imidazolyl (e.g., 2H-imidazol-2-yl and 2H-imidazol-4-yl), pyrazolyl (e.g., 1H-pyrazol-3-yl, 1H-pyrazol-4-yl, and 1H-pyrazol-5-yl), pyridyl (e.g., pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl), pyrimidinyl (e.g., pyrimidine- -2-yl, pyrimidin-4-yl, and pyrimidin-5-yl), thiazolyl (e.g., thiazol-2-yl, thiazol-4-yl, and thiazol-5-yl), isothiazolyl (e.g., isothiazol-3-yl, isothiazol-4-yl, and isothiazol-5-yl), oxazolyl (e.g., oxazol-2-yl, oxazol-4-yl, and oxazol-5-yl), isoxazolyl (e.g., isoxazol-3-yl, isoxazol-4-yl, and isoxazol-5-yl), or indazolyl (e.g., 1H-indazol-3-yl). The term "heteroaryl" also includes possible N-oxides. A non-limiting example of an N-oxide is pyridyl N-oxide.

[0190] In one embodiment, the heteroaryl is a 5- or 6-membered heteroaryl. In one embodiment, the heteroaryl is a 5-membered heteroaryl, i.e., the heteroaryl is a monocyclic aromatic ring system having 5 ring atoms in which at least one carbon atom of the ring is replaced with a heteroatom independently selected from nitrogen, oxygen, and sulfur. Non-limiting exemplary 5-membered heteroaryl groups include thienyl, furyl, pyrrolyl, oxazolyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, and isoxazolyl.

[0191] In another embodiment, the heteroaryl is a 6-membered heteroaryl, e.g., the heteroaryl is a monocyclic aromatic ring system having 6 ring atoms in which at least one carbon atom of the ring is replaced with a nitrogen atom. Non-limiting exemplary 6-membered heteroaryl groups include pyridyl, pyrazinyl, pyrimidinyl, and pyridazinyl.

[0192] In one embodiment, the group or point of attachment of the heteroaryl is on the aromatic ring and can be either a carbon atom or a heteroatom.

[0193] In this disclosure, the term "C-linked heteroaryl" or "carbon-linked heteroaryl," as used by itself or as part of another group, refers to a heteroaryl attached to the remainder of the molecule at any available carbon atom on the heteroaryl ring.

[0194] In this disclosure, the term "optionally substituted heteroaryl," used by itself or as part of another group, refers to heteroaryl that is either unsubstituted or substituted with 1 to 4 substituents, such as 1 or 2 substituents independently selected from the group consisting of halo, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido, sulfonamido, alkylcarbonyl, arylcarbonyl, alkylsulfonyl, arylsulfonyl, carboxy, carboxyalkyl, optionally substituted alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclo, (alkoxy)alkyl, (amino)alkyl, (carboxamido)alkyl, mercaptoalkyl, and (heterocyclo)alkyl. In one embodiment, the optionally substituted heteroaryl has one substituent. Any available carbon or nitrogen atom can be substituted. Non-limiting exemplary substituted heteroaryl groups include, but are not limited to: [ka] Includes:

[0195] The term "optionally substituted heteroaryl" encompasses a heteroaryl group fused to an optionally substituted cycloalkyl or fused to an optionally substituted heterocyclo group. An optionally substituted heteroaryl fused to an optionally substituted cycloalkyl or fused to an optionally substituted heterocyclo group may be attached to the rest of the molecule at any available carbon atom on the heteroaryl ring. Non-limiting examples include: [ka] Examples include:

[0196] In this disclosure, the term "halogen" is intended to include fluorine, chlorine, bromine and iodine.

[0197] In the present disclosure, the term "internucleoside linkage" refers to a naturally occurring or modified linkage between two adjacent nucleosides in an oligonucleotide or CDN. Naturally occurring RNA and DNA contain phosphorodiester internucleoside linkages. An example of a modified internucleoside linkage is a phosphorothioate linkage.

[0198] In the present disclosure, the term "chimeric nucleic acid" or "chimeric sequence" refers to a nucleic acid having different internucleoside linkages. In one embodiment, the chimeric nucleic acid has a combination of phosphorothioate and phosphonate linkages.

[0199] In the present disclosure, the term "heterochiral nucleic acid" refers to a nucleic acid containing internucleoside linkages containing phosphorus atoms in different stereochemical configurations. Similarly, the term "homochiral nucleic acid" refers to a nucleic acid containing internucleoside linkages containing phosphorus atoms in the same stereochemical configuration.

[0200] In the present disclosure, the term "protecting group" refers to a group that protects functional groups such as alcohols, amines, carbonyls, carboxylic acids, phosphates, terminal alkynes, and the like, from undesired chemical reactions. In some embodiments, the functional group is a nucleophilic group. Examples of alcohol protecting groups include, but are not limited to, acetyl (Ac), benzoyl (Bz), benzyl (Bn), β-methoxyethoxymethyl ether (MEM), dimethoxytrityl (DMT), methoxymethyl ether (MOM), methoxytrityl (MMT), p-methoxybenzyl ether (PMB), trimethylsilyl (TMS), tert-butyldimethylsilyl (TBS), tert-butyldiphenylsilyl ether (TBDPS), tri-isopropylsilyloxymethyl (TOM), trityl (triphenylmethyl, Tr), pivaloyl (Piv), and the like. In one embodiment, the protecting group is 4,4'-dimethoxytrityl. Examples of amine protecting groups include, but are not limited to, carbobenzyloxy (Cbz), isobutyryl (iBu), p-methoxybenzyl carbonyl (MOZ), tert-butylcarbonyl (Boc), acetyl (Ac), benzoyl (Bz), benzyl (Bn), p-methoxybenzyl (PMB), p-methoxyphenyl (PMP), tosyl (Ts), and the like. Examples of carbonyl protecting groups include, but are not limited to, acetals and ketals, acylals, dithianes, and the like. Examples of carboxylic protecting groups include, but are not limited to, methyl esters, benzyl esters, tert-butyl esters, silyl esters, orthoesters, oxazolines, and the like. Examples of phosphate protecting groups include, but are not limited to, 2-cyanoethyl, methyl, and the like. Examples of terminal alkyne protecting groups include, but are not limited to, propargyl and silyl groups. In one embodiment, a protecting group is used to protect the 5'-hydroxy group of a nucleoside used in the methods of the present disclosure. In one embodiment, the protecting group is DMT. In another embodiment, a protecting group is used to protect the nucleobase of a nucleoside used in the methods of the present disclosure. In some embodiments, the protecting group is an amine protecting group. In one embodiment, the protecting group is Ac. In another embodiment, the protecting group is Bz.In yet another embodiment, the protecting group is iBu.

[0201] [Example] Useful embodiments of the compounds / reagents and methods of the present disclosure are provided in the following examples, which should be understood as being given solely to illustrate the invention.

[0202] General Experiment Tetrahydrofuran (THF), N,N-dimethylformamide (DMF), dichloromethane (DCM), acetonitrile (MeCN), and methanol (MeOH) were obtained by passing previously degassed solvents through activated alumina columns. DBU was purchased from Chem-Impex. All 5'-DMTr-protected nucleosides with conventional nucleobase protection (T = none, A = benzoyl, G = isobutyryl, C = benzoyl) were purchased from Chem-Impex. All reagents were purchased of the highest commercially available quality and used without purification unless otherwise stated. Yields were based on chromatographic and spectroscopic ( 1 The term refers to homogeneous material (H NMR). Reactions were monitored via thin-layer chromatography (TLC), GC / MS, GC / FID, or LC / MS. TLC was performed using E. Merck 0.25 mm silica plates (60F-254) with short-wave UV light as the visualization agent and phosphomolybdic acid, p-anisaldehyde, or KMnO4 as the developer, and heat. NMR spectra were recorded on Bruker DRX-600, DRX-500, and AMX-400 instruments. Residual deuterated solvents (CHCl3, CHCl2, DMSO, MeOH, acetone) were removed by NMR. 1 H NMR showed 7.26, 5.32, 2.50, 3.31 and 2.05 ppm, respectively. 13C NMR was calibrated using 77.16, 53.84, 39.52, 49.00, and 29.84 ppm, respectively. The following abbreviations were used to describe the multiplicities: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, and br = broad. Column chromatography was performed using E. Merck silica gel (60, particle size 0.043–0.063 mm), and preparative TLC (pTLC) was performed on Merck silica plates (60F-254). High-resolution mass spectra (HRMS) were recorded on an Agilent LC / MSD TOF mass spectrometer using electrospray ionization time-of-flight reflectron experiments. Melting points were recorded on a Fisher-Johns 12-144 melting point apparatus and were uncorrected. Enantiomeric ratios were determined using a Waters UPC2 SFC or Agilent Technologies 1220 Infinity II LC HPLC equipped with a photodiode array detector. Optical rotation data were recorded on an Anton Paar 100 Modular Circular Polarimeter.

[0203] Example 1 Synthesis of PI reagent [ka] a. Synthesis of SI-1 and SI-2 Compounds SI-1 and SI-2 (both enantiomers) were synthesized according to published procedures, see Knouse et al., Unlocking P(V): Reagents for chiral phosphorothioate synthesis, Science 2018, 361, 1234. [ka]

[0204] [ka] [ka] [ka] The synthesis of SI-1 follows a similar procedure to the previously reported synthesis of the Ψ-reagent, which can be illustrated with a detailed schematic guide. See Knouse et al., Unlocking P(V): Reagents for chiral phosphorothioate synthesis, Science 2018, 361, 1234. To a solution of SI-1 (1.00 g, 1.68 mmol, 1.0 equiv.) and trans-(+)-limonene oxide SI-2 (0.383 g, 2.52 mmol, 1.5 equiv.) in dichloromethane (5.0 mL) was added trifluoroacetic acid (0.19 mL, 2.52 mmol, 1.5 equiv.). The reaction was allowed to warm to 35 °C and stir for 1 h. The reaction mixture was cooled to ambient temperature, and hexane (10 mL) was added in batches to form a biphasic mixture. The stream was washed with water (5 mL), saturated NaHCO3 (10 mL), and KH2PO4 (10% aqueous solution, 3 mL). The organic phase was filtered through a MgSO4 pad and concentrated to approximately 3 mL. Methanol (5 mL) was added and the batch was concentrated to approximately 3 mL; this operation was repeated twice. The mixture was cooled to 5-10°C and stirred for 5 minutes. The resulting slurry was filtered, and the reactor and cake were washed with cold methanol (1 mL). The filter cake was dried under vacuum and [ka] was obtained as a crystalline white solid (0.56 g, 75% yield, >20:1 dr, >98:2 er). Physical state: white crystalline solid; melting point = 104-106℃; [α] 25 D =-125°(c=1.00 in CHCl3);R f =0.50 (hexane:toluene = 1:1) 1H NMR (600MHz, CDCl3) δ 4.96(s,1H), 4.70(d,J=2.1Hz,1H), 2.90(dd,J=13.4, 2.9Hz,1H), 2.42(s,1H), 2.10(ddp,J=13.7, 6.4, 2.2Hz,2H ), 2.02(ddd,J=12.5, 4.3, 2.7Hz,1H), 1.77-1.66(m,2H), 1.65(s,3H), 1.64(s,3H), 1.52(td,J=13.3, 4.2Hz,1H) 13 C NMR (151MHz, CDCl3) δ 148.86, 147.19, 144.69, 144.02, 142.28, 138.75, 137.05, 111.83, 104.30, 93.6 4, 93.60, 58.65, 58.64, 38.81, 34.21, 34.13, 28.39, 28.32, 24.89, 22.18, 18.74 19 F NMR (376MHz, CDCl3) δ -128.54(dd,J=20.7, 4.6Hz,2F), -146.87(ddd,J=25.5, 14.7, 6.0Hz,1F), -159.16--160.51(m,2F) 31 P NMR(162MHz, CDCl3) δ 102.08 HRMS(ESI-TOF):C 16 H 17 Calculated value [M+H] for F5OPS3 + : 447.0094, Measurement value: 447.0093

[0205] [ka] [ka] The synthesis of 1b was carried out by the same procedure as described for 1a. All characterizing data are 25 D = +124° (c = 1.00 in CHCl3).

[0206] Example 2 Synthesis of adamantylmethanol-loaded phosphonothioates [ka] A flame-dried culture tube under argon was charged with 1-adamantanemethanol (16.6 mg, 0.1 mmol, 1.0 equiv.), 12a (52.5 mg, 0.2 mmol, 2.0 equiv.), and MeCN (1 mL). DBU (0.05 mL, 3.0 mmol, 3.0 equiv.) was added, and the resulting solution was stirred at room temperature for 4 h. 2 mL of 1x PBS solution was added, and the mixture was extracted with DCM (3x3 mL). The organics were combined, washed with brine (1 mL), dried over Na2SO4, filtered, and concentrated. The resulting crude oil was dissolved in THF (1 mL). Et3N (2 equiv.) and BnBr (2 equiv.) were added, and the resulting mixture was stirred for 12 h. The reaction was quenched with 1 mL of water and extracted with EtOAc (3x3 mL). The organics were combined, dried over Na2SO4, filtered, and concentrated. Purification by silica gel chromatography (hexane / EtOAc=5:2) afforded 24.0 mg (69%) of the title compound SI-3. Physical state: colorless oil; R f =0.40 (hexane / EtOAc = 5:2) 1 H NMR (400MHz, CDCl3) δ 7.41-7.22(m,5H), 4.14-3.97(m,2H), 3.65(dd,J=9.6, 6.5Hz,1H), 3.46(d d,J=9.7, 6.8Hz,1H), 1.98(s,3H), 1.77-1.57(m,9H), 1.50(d,J=2.9Hz,6H) 13 C NMR (151MHz, CDCl3) δ 138.13, 138.11, 129.05, 128.82, 127.64, 74.81, 74.76, 39.02, 37.03, 34.68, 34.65, 33.68, 33.63, 28.10, 20.35, 19.62 31 P NMR(162MHz, CDCl3) δ 53.64 HRMS(ESI-TOF):C 19 H28 Calculated value for O2PS [M+H] + :351.1548, Measurement value:351.1546

[0207] Example 3 Synthesis of Grignard and organolithium reagents a. Synthesis of Grignard reagents All Grignard reagents were prepared from aryl bromides via Mg insertion in the presence of LiCl or purchased from commercial sources.

[0208] 1. General procedure for Mg insertion: LiCl (0.530 g, 12.5 mmol, 1.25 equiv.) was flame-dried under vacuum. After cooling, the flask was placed under an Ar atmosphere, and Mg turnings (0.608 g, 25 mmol, 2.5 equiv.), iodine (2-3 grains), and THF (2 mL) were added. In a separate flask under argon, a solution of aryl bromide (10 mmol, 1.0 equiv.) in THF (8 mL) was prepared. A few drops of the aryl bromide solution in THF were added to the flask containing the Mg and gently heated with a heat gun until the solution changed color from brown to colorless. The remaining aryl bromide solution was then slowly added. After complete addition, the mixture was placed in an oil bath (55 °C) and heated for 1-2 h. The Grignard was titrated against iodine / LiCl prior to use according to the published knochel procedure.

[0209] The following Grignard reagents were synthesized using the above procedure: [ka]

[0210] The following Grignard reagents were purchased directly from Sigma-Aldrich: [ka]

[0211] b. Synthesis of Grignard Reagents Unless otherwise noted, all organolithium reagents were either freshly prepared via direct deprotonation of terminal alkynes by lithium-halogen exchange of aryl bromides or purchased from commercial sources. FcLi was prepared according to published procedures. See Han, ZS et al., Efficient Asymmetric Synthesis of P-Chiral Phosphine Oxides via Properly Designed and Activated Benzoxazaphosphinine-2-oxide Agents. J. Am. Chem. Soc. 2013, 135, 2474-2477.

[0212] 1. General procedure for lithium-halogen exchange: A flame-dried round-bottom flask under an argon atmosphere was charged with aryl bromide (11 mmol, 1.1 equiv.) and THF (40 mL). The mixture was cooled to -78 °C, and nBuLi solution (10 mmol, 1.0 equiv.) was added dropwise. The resulting mixture was stirred at -78 °C for 30 min and used as is.

[0213] The following organolithium reagents were synthesized using the methods described above: [ka]

[0214] Direct deprotonation of terminal alkynes follows the same procedure as lithium-halogen exchange. The following organolithium reagents have been synthesized by direct deprotonation: [ka]

[0215] 2. Synthesis of FcLi: [ka] A flame-dried round-bottom flask under an argon atmosphere was charged with ferrocene (186 mg, 1 mmol, 1.0 equiv.), tBuOK (0.15 mL, 1.0 M, 0.15 mmol, 0.15 equiv.), and THF (8 mL). The reaction was cooled to -78 °C, and tBuLi (1.18 mL, 1.7 M, 2 mmol, 2.0 equiv.) was added dropwise. The resulting mixture was allowed to stir at -78 °C for 1 h, held at 0 °C for 30 min, and then used as is.

[0216] The following organolithium reagents were purchased from Sigma-Aldrich: [Table 9]

[0217] Example 4 Synthesis of P-chiral phosphines a. Loading [ka] 1. General operations A A flame-dried 250 mL round-bottom flask was charged with 1 (4.46 g, 10 mmol, 1 equiv.). The flask was evacuated and backfilled with argon, after which anhydrous THF (100 mL, 0.1 M) was introduced via syringe. Grignard reagent (20 mmol, 2 equiv.) was then added dropwise, and the resulting solution was 31 The mixture was allowed to stir at room temperature until P NMR indicated complete consumption of the starting material. The reaction was quenched by the slow addition of saturated aqueous NH4Cl (20 mL) and diluted with water (80 mL) and EtOAc (150 mL). The two layers were separated, and the aqueous layer was washed twice with EtOAc (2 x 80 mL). The organic layers were combined, washed with saturated aqueous NaHCO3 (50 mL), brine (50 mL), and dried over anhydrous Na2SO4. The mixture was then filtered, concentrated in vacuo, and purified by silica gel chromatography to give the desired product.

[0218] 2. Compound 8a: [ka] 1a and Grignard reagent SI-10 were treated according to general procedure A on a 10 mmol scale with a reaction time of 2 h. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 1.93 g (60%, >20:1 dr) of the title compound 8a. On a 50 mmol scale, 9.20 g (57%, >20:1 dr) of 8a was obtained after purification. Physical state: white solid; melting point = 108-110℃; [α] 25 D =-27.3°(c=1.00 in CHCl3);R f =0.35 (hexane / toluene = 1:2) 1 H NMR (400MHz, CDCl3) δ 7.87(ddd,J=15.2, 8.1, 1.5Hz,2H), 7.56-7.40(m,3H), 4.98(s,1H), 4.89(s,1H), 3.86(dd,J= 13.2, 3.1Hz,1H), 2.50(s,1H), 2.26-2.09(m,3H), 2.08-1.84(m,2H), 1.83(s,3H), 1.73(s,3H) 13 C NMR (126MHz, CDCl3) δ 145.45, 138.93, 138.02, 132.21, 132.19, 130.30, 130.20, 128.70, 128.58, 111.94, 92.62, 92.59, 57.25, 57.24, 39.00, 34.91, 34.82, 29.05, 28.98, 25.36, 22.83, 19.33 31 P NMR(162MHz, CDCl3) δ 97.21 HRMS(ESI-TOF):C 16 H 22 As OPS2, the calculated value [M+H] + :325.0850, Measurement value:325.0849

[0219] 3. Compound 8b: [ka] 1b was treated with Grignard reagent SI-10 according to general procedure A on a 10 mmol scale with a reaction time of 2 h. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 2.11 g (65%, >20:1 dr) of the title compound 8b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = +31.6° (c = 1.00 in CHCl3).

[0220] 4. Compound 12a: [ka] 1a was treated with Grignard reagent SI-11 on a 10 mmol scale for a reaction time of 2 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene / EtOAc = 1:1:0 to 1:2:0 to 1:2:0.05) gave 1.73 g (66%, >20:1 dr) of the title compound 12a. Physical state: white solid; melting point = 102-105℃; [α] 25 D =+136.4°(c=0.50 in CHCl3);R f =0.20 (hexane / toluene = 1:2) 1 H NMR (400MHz, CDCl3) δ 5.01(s,1H), 4.89(s,1H), 3.74(dd,J=13.3, 3.1Hz,1H), 2.48(s,1H), 2.23(d,J=13.8Hz,1H), 2.20(s,3H), 2.1 7(d,J=0.8Hz,3H), 2.11(d,J=11.2Hz,1H), 2.06-2.00(m,1H), 1.80(d,J=8.1Hz,3H), 1.76(s,3H), 1.68(s,3H) 13 C NMR (151MHz, CDCl3) δ 145.25, 112.07, 92.26, 92.23, 56.99, 56.97, 38.93, 35.16, 35.08, 31.27, 30.74, 28.99, 28.93, 25.21, 22.78, 18.85 31 P NMR(162MHz, CDCl3) δ 106.19 HRMS(ESI-TOF):C 11 H 20 As OPS2, the calculated value [M+H] + :263.0693, Measurement value:263.0697

[0221] 5. Compound 12b: [ka] 1b and Grignard reagent SI-11 were treated according to general procedure A on a 5 mmol scale with a reaction time of 2 h. Purification by silica gel column chromatography (hexane / toluene / EtOAc = 1:1:0 to 1:2:0 to 1:2:0.05) afforded 923 mg (70%, >20:1 dr) of the title compound 12b. On a 25 mmol scale, 3.41 g (52%, >20:1 dr) of 12b was obtained after purification. All physical and spectroscopic properties indicated the optical rotation was [α]. 25 D = -130.2° (c = 0.50 in CHCl3).

[0222] 6. Compound 13a: [ka] 1a and Grignard reagent SI-5 were treated according to general procedure A on a 0.5 mmol scale for a reaction time of 12 h. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 60 mg (33%, >20:1 dr) of the title compound 13a. Physical state: colorless gel; [α] 25 D =-25.0°(c=0.70 in CHCl3);R f =0.41 (Hexane / EtOAc = 92:8) 1H NMR (400MHz, CDCl3) δ 7.51(dd,J=15.2, 1.8Hz,1H), 7.23-7.09(m,2H), 4.85(d,J=1.4Hz,1H), 4.78(s,1H), 3.55(ddd,J=13.3, 3.2, 1.6Hz, 1H), 2.70(s,3H), 2.44(t,J=5.3Hz,1H), 2.33(s,3H), 2.23-2.06(m,3H), 2.02-1.81(m,3H), 1.79(s,3H), 1.66(s,3H) 13 C NMR (151MHz, CDCl3) δ 145.29, 137.41, 136.73, 136.68, 136.63, 135.04, 134.95, 132.36, 132.34, 132.29, 132.19, 129.44, 129.38, 111.88, 92.48, 9 2.44, 77.37, 77.16, 77.08, 76.95, 56.19, 56.17, 38.89, 34.78, 34.71, 28.96, 28.89, 25.35, 22.78, 21.56, 21.53, 21.13, 19.69 31 P NMR(162MHz, CDCl3) δ 94.99 HRMS(ESI-TOF):C 18 H 26 As OPS2, the calculated value [M+H] + :353.1163, Measurement:353.1168

[0223] 7. Compound 13b: [ka] 1b was treated with Grignard reagent SI-5 on a 0.5 mmol scale for 12 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 58 mg (32%, >20:1 dr) of the title compound 13b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = +31° (c = 0.60 in CHCl3).

[0224] 8. Compound 14a: [ka] 1a was treated with Grignard reagent SI-6 on a 1 mmol scale for 12 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) afforded 292 mg (77%, >20:1 dr) of the title compound 14a. Physical state: pale yellow solid; melting point = 114-116℃; [α] 25 D =-121.8°(c=1.00 in CHCl3); R f =0.24 (Hexane / Et2O=9:1) 1 H NMR (500MHz, CDCl3) δ 8.84(dd,J=8.4, 0.9Hz,1H), 8.03-7.83(m,3H), 7.65(ddd,J=8.5, 6.9, 1.4Hz,1H), 7.56( ddd,J=8.1, 6.9, 1.2Hz,1H), 7.47(ddd,J=8.1, 7.3, 3.6Hz,1H), 4.80(d,J=1.4Hz,1H), 4. 73(s,1H), 3.57(ddd,J=13.3, 3.2, 1.6Hz,1H), 2.44(d,J=6.0Hz,1H), 2.22(ddd,J=12.4, 4.4, 2.8Hz,1H), 2.16-2.08(m,2H), 2.02-1.78(m,3H), 1.86(s,3H), 1.62(d,J=1.3Hz,3H) 13C NMR (151MHz, CDCl3) δ 163.86, 163.74, 159.73, 159.72, 145.52, 145.34, 145.15, 136.41, 136.32, 134.58, 133. 59, 131.03, 130.99, 129.45, 129.43, 129.17, 129.08, 128.80, 127.79, 114.09, 114.07, 1 14.03, 97.12, 87.93, 87.76, 87.69, 86.90, 86.86, 85.92, 85.89, 75.87, 75.84, 72.60, 65.22, 65.18, 63.37, 55.55, 42.11, 40.79, 40.77, 26.16, 18.50, 12.25, 11.30, -4.52, -4.65 31 P NMR(162MHz, CDCl3) δ 92.77 HRMS(ESI-TOF):C 20 H 24 As OPS2, the calculated value [M+H] + :375.1006, Measurement:375.1006

[0225] 9. Compound 14b: [ka] 1b was treated with Grignard reagent SI-6 on a 0.5 mmol scale for 12 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 152 mg (80%, >20:1 dr) of the title compound 14b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = +125.1° (c = 0.35 in DCM).

[0226] 10. Compound 15a: [ka] 1a and Grignard reagent SI-4 were treated according to general procedure A on a 0.5 mmol scale for a reaction time of 12 h. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 74 mg (37%, >20:1 dr) of the title compound 15a. Physical state: pale yellow gel [α] 25 D =-83.8°(c=0.22 in DCM) R f =0.31 (Hexane / Et2O=9:1) 1 H NMR (400MHz, CDCl3) δ 7.44(dd,J=7.6, 1.4Hz,1H), 7.40-7.27(m,8H), 4.95(d,J=1.4Hz,1H), 4.89-4.82(m,1H), 3.74(ddd,J=1 3.3, 3.2, 1.6Hz,1H), 2.50(s,1H), 2.25-2.10(m,3H), 1.97-1.77(m,3H), 1.75(s,3H), 1.74-1.71(m,3H) 13 C NMR (151MHz, CDCl3) δ 145.07, 143.68, 140.20, 131.89, 131.05, 129.01, 128.55, 128.41, 128.25, 127.68, 112. 31, 93.62, 93.59, 56.99, 56.97, 38.84, 34.76, 34.68, 28.76, 28.69, 25.31, 22.78, 19.60 31 P NMR(162MHz, CDCl3) δ 77.83 HRMS(ESI-TOF):C 22 H 26 As OPS2, the calculated value [M+H] + : 401.1163, Measurement: 401.1157

[0227] 11. Compound 15b: [ka] 1b was treated with Grignard reagent SI-4 on a 4 mmol scale for 2 h reaction time according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 882 mg (55%, >20:1 dr) of the title compound 15b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = +95.7° (c = 1.00 in CHCl3).

[0228] 12. Compound 16a: [ka] 1a and Grignard reagent SI-7 were treated according to general procedure A on a 0.5 mmol scale with a reaction time of 12 h. Purification by silica gel column chromatography (hexane / toluene = 2:1) gave 126 mg (68%, >20:1 dr) of the title compound 16a. Physical state: pale yellow gel [α] 25 D =-48.8°(c=0.50 in CHCl3) R f =0.2 (hexane / toluene = 2:1) 1 H NMR (400MHz, CDCl3) δ 8.41(d,J=17.9Hz,1H), 7.95-7.81(m,4H), 7.63-7.53(m,2H), 4.99(d,J=1.4Hz,1H), 4.92(s,1H), 3.95(dd,J=13.3, 3.1 Hz,1H), 2.51(s,1H), 2.26-2.15(m,3H), 2.09(td,J=13.3, 12.6, 4.1Hz,1H), 1.99-1.88(m,2H), 1.87(s,3H), 1.73(s,3H) 13C NMR (151MHz, CDCl3) δ 144.93, 135.15, 134.39, 134.31, 134.29, 131.82, 131.71, 131.46, 131.38, 128.80, 128.20, 128.10, 127.93, 127.29, 126.61, 124.81, 124.72, 111.35, 92.14, 92.11, 56.73, 56.72, 38.47, 34.38, 34.31, 28.49, 28.43, 24.82, 22.26, 18.84 31 P NMR(162MHz, CDCl3) δ 97.21 HRMS(ESI-TOF):C 20 H 24 As OPS2, the calculated value [M+H] + :375.1006, Measurement value:375.0999

[0229] 13. Compound 16b: [ka] 1b was treated with Grignard reagent SI-7 on a 0.5 mmol scale for 12 h reaction time according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1) gave 137 mg (72%, >20:1 dr) of the title compound 16b. All physical and spectroscopic properties were consistent with the optical rotation of [α] 25 D = +40.1° (c = 0.21 in DCM).

[0230] 14. Compound 17a: [ka] 1a was treated with Grignard reagent SI-13 on a 2 mmol scale for 12 h reaction time according to general procedure A. Purification by silica gel column chromatography (hexane / toluene / EtOAc = 2:1:0 to 1:1:0 to 1:1:0.05) gave 420 mg (59%, >20:1 dr) of the title compound 17a. Physical state: colorless oil [α] 25 D =-38.2° (c=0.50 in DCM) R f =0.22(ヘキサン / Et2O=9:1) 1 H NMR (400MHz, CDCl3) δ 7.98(ddd,J=17.3,7.7,1.7Hz,1H),7.52-7.42(m,1H),7.01(tdd,J=7.6,3. 4. 1.0Hz, 1H), 6.95 (ddd, J=8.2, 7.1, 0.9Hz, 1H), 4.95 (q, J=1.4Hz, 1H), 4.8 8(s,1H), 3.98(ddd,J=13.5, 3.1, 0.7Hz,1H), 3.92(s,3H), 2.48(s,1H), 2.2 5-2.07(m,3H), 2.00-1.84(m,3H), 1.82(s,3H), 1.73(dt,J=1.4, 0.7Hz,3H) 13 C NMR (151MHz, CDCl3) δ 159.92, 159.89, 145.68, 134.24, 134.22, 134.07, 134.01, 125.12, 124.37, 120.49, 120.39, 111.99, 111 .94, 111.88, 92.25, 92.22, 56.65, 56.64, 55.93, 39.05, 34.93, 34.85, 28.82, 28.76, 25.26, 22.78, 19.49 31 P NMR (162MHz, CDCl3) δ 93.72 HRMS (ESI-TOF):C 17 H 24 O2PS2として, calculation value [M+H] + :355.0955, measured value: 355.0951

[0231] 15. Compound 17b:

change

[0232] 16. Compound 18a: [ka] 1a was treated with Grignard reagent SI-15 on a 1 mmol scale for 0.5 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:1) gave 146 mg (54%, >20:1 dr) of the title compound 18a. Physical state: colorless oil [α] 25 D =+12.5°(c=0.28 in CHCl3) R f =0.41 (Hexane / Et2O=9:1) 1 H NMR (500MHz, CDCl3) δ 6.38(ddd,J=29.8, 17.7, 11.4Hz,1H), 6.20(ddd,J=29.4, 17.7, 1.3Hz,1H), 5.98( ddd,J=57.5, 11.4, 1.3Hz,1H), 4.99(d,J=1.5Hz,1H), 4.88(s,1H), 3.74(dd,J=13. 3, 3.2Hz,1H), 2.48(s,1H), 2.22(dd,J=13.8, 1.8Hz,1H), 2.16-2.09(m,1H), 2.06( dd,J=7.3, 4.7Hz,1H), 1.92-1.77(m,3H), 1.75(dt,J=1.5, 0.8Hz,3H), 1.72(s,3H) 13C NMR (151MHz, CDCl3) δ 163.86, 163.74, 159.73, 159.72, 145.52, 145.34, 145.15, 136.41, 136.32, 134.58, 133. 59, 131.03, 130.99, 129.45, 129.43, 129.17, 129.08, 128.80, 127.79, 114.09, 114.07, 1 14.03, 97.12, 87.93, 87.76, 87.69, 86.90, 86.86, 85.92, 85.89, 75.87, 75.84, 72.60, 65.22, 65.18, 63.37, 55.55, 42.11, 40.79, 40.77, 26.16, 18.50, 12.25, 11.30, -4.52, -4.65 31 P NMR(162MHz, CDCl3) δ 93.96 HRMS(ESI-TOF):C 12 H 20 As OPS2, the calculated value [M+H] + :275.0693, Measurement:275.0695

[0233] 17. Compound 18b: [ka] 1b was treated with Grignard reagent SI-15 on a 0.5 mmol scale for 0.5 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:1) gave 85 mg (63%, >20:1 dr) of the title compound 18b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = -7.96° (c = 0.21 in DCM).

[0234] 18. Compound 19a: [ka] 1a was treated with Grignard reagent SI-14 on a 1 mmol scale for 1 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 232 mg (67%, >20:1 dr) of the title compound 19a. Physical state: pale yellow solid Melting point = 107-110℃ [α] 25 D =+115.3°(c=0.22 in DCM) R f =0.24 (Hexane / Et2O=9:1) 1 H NMR (500MHz, CDCl3) δ 7.57-7.49(m,2H), 7.48-7.40(m,1H), 7.40-7.33(m,2H), 5.02(d,J=1.5Hz, 1H), 4.95(s,1H), 4.14(dt,J=13.2, 2.7Hz,1H), 2.51(t,J=6.0Hz,1H), 2.29( ddd,J=13.8, 3.4, 1.7Hz,1H), 2.15(d,J=15.0Hz,1H), 2.10(ddd,J=12.5, 4.4 , 2.9Hz,1H), 2.03-1.86(m,2H), 1.85-1.79(m,1H), 1.78(s,3H), 1.67(s,3H) 13 C NMR (151MHz, CDCl3) δ 145.31, 132.66, 132.65, 130.72, 128.62, 120.06, 120.03, 112.15, 99.29, 99.03, 93.04, 9 3.01, 87.23, 85.86, 57.13, 57.10, 39.01, 35.14, 35.06, 29.03, 28.96, 25.49, 22.86, 19.79 31 P NMR(162MHz, CDCl3) δ 60.28 HRMS(ESI-TOF):C 18 H 22 As OPS2, the calculated value [M+H] + :349.0850, Measurement value:349.0841

[0235] 19. Compound 19b: [ka] 1b was treated with the Grignard reagent SI-14 on a 0.5 mmol scale for 1 h according to general procedure A. Purification by silica gel column chromatography (hexane / toluene = 2:1 to 1:2) gave 118 mg (68%, >20:1 dr) of the title compound 19b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = -104.0° (c = 0.20 in DCM).

[0236] b. Coupling 1. General Procedure B (One-pot MeI Quench): [ka] An organolithium reagent (4.0 mmol, 2.0 equiv., see Example 3, "Synthesis of Grignard and Organolithium Reagents") in 14 mL of THF was cooled to -78°C if not already at that temperature. A solution of the starting material (2.0 mmol, 1.0 equiv.) in THF (6 mL) was prepared in a flame-dried flask under argon and then added dropwise to the flask containing the organolithium reagent. The resulting mixture was stirred for 3 hours while maintaining the temperature at -78°C. A small aliquot of 31 After P NMR analysis showed that the starting material was completely consumed, the reaction was quenched with excess methyl iodide (8.0 mmol, 4.0 equiv.), allowed to warm to room temperature, and stirred for another hour. To the resulting mixture was added saturated aqueous NH4Cl (20 mL) and EtOAc (40 mL). The layers were separated, and the aqueous layer was washed with EtOAc (2 x 20 mL). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography to give the desired product.

[0237] 2. General operation C (step type): [ka] An organolithium reagent (4.0 mmol, 2.0 equiv., see Example 3, "Synthesis of Grignard and Organolithium Reagents") in 14 mL of THF was cooled to -78°C if not already at that temperature. A solution of the starting material (2.0 mmol, 1.0 equiv.) in THF (6 mL) was prepared in a flame-dried flask under argon and then added dropwise to the flask containing the organolithium reagent. The resulting mixture was stirred for 3 hours while maintaining the temperature at -78°C. A small aliquot of 31 After P NMR analysis showed complete consumption of the starting material, the reaction was carefully quenched by slow addition of saturated aqueous NH4Cl (20 mL) and then diluted with water (10 mL) and DCM (40 mL). The layers were separated, and the aqueous layer was washed with DCM (2 x 20 mL). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was dissolved in THF (10 mL), and to it was added Et3N (4.0 mmol, 2.0 equiv.) and MeI (4.0 mmol, 2.0 equiv.). After stirring for 1 h, the reaction was diluted with water (20 mL) and extracted with EtOAc (3 x 40 mL). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography to give the desired product.

[0238] 3. Compound 11a 8a and the organolithium reagent SI-16 were treated on a 4 mmol scale according to general procedure B. Purification by silica gel column chromatography (hexane / EtOAc = 2:1 to 1:2) gave 916 mg (82%, >98:2 er) of the title compound 11a. On a 20 mmol scale, 4.70 g (84%, >98:2 er) of 11a was obtained after purification. Physical state: pale yellow solid Melting point = 89-92℃ [α] 25 D =-13.1°(c=1.00 in CHCl3) R f =0.42 (Hexane / EtOAc = 1:2) 1 H NMR (400MHz, CDCl3) δ 7.97(dddd,J=14.2, 7.6, 1.8, 0.4Hz,1H), 7.94-7.86(m,2H), 7.55-7.40(m,4H), 7.08(tdd,J =7.5, 2.5, 0.9Hz,1H), 6.89(ddd,J=8.3, 6.1, 0.9Hz,1H), 3.71(s,3H), 2.27(d,J=12.3Hz,3H) 13 C NMR (126MHz, CDCl3) δ 160.45, 160.41, 134.73, 134.55, 134.53, 134.01, 133.95, 133.85, 131.89, 131.86, 131.53, 1 31.44, 128.36, 128.25, 121.66, 120.99, 120.89, 120.82, 111.66, 111.60, 55.57, 10.43, 10.41 31 P NMR(162MHz, CDCl3) δ 41.34 HRMS(ESI-TOF):C 14 H 16 Calculated value for O2PS [M+H] + :279.0609, Measurement:279.0612

[0239] 4. Compound 11b [ka] 8b was treated with the organolithium reagent SI-16 on a 5 mmol scale according to General Procedure B. Purification by silica gel column chromatography (hexane / EtOAc = 2:1 to 1:2) gave 1.07 g (77%, >98:2 er) of the title compound 11b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D Identical to 11a except that c = +15.6 (c = 1.00 in CHCl3).

[0240] 5. Compound 20a [ka] 8a and the organolithium reagent SI-17 were treated on a 0.5 mmol scale according to general procedure B. Purification by silica gel column chromatography (hexane / EtOAc = 3:1 to 1:1) afforded 72.0 mg (52%, >98:2 er) of the title compound 20a. Physical state: White amorphous solid [α] 25 D =+15.5°(c=0.40 in CHCl3) R f =0.45 (hexane / EtOAc = 1:2) 1 H NMR (400MHz, CDCl3) δ 7.85-7.76(m,2H), 7.65(dd,J=14.8, 8.3Hz,1H), 7.57-7.50(m,1H), 7.50-7.43(m,2H) ), 7.07(ddd,J=5.5, 1.6, 0.8Hz,2H), 2.49(s,1H), 2.35(s,4H), 2.27(d,J=11.9Hz,3H) 13 C NMR (126MHz, CDCl3) δ 143.15, 143.13, 142.59, 142.51, 134.02, 133.17, 133.14, 133.12, 133.04, 133.02, 132.22, 132.20, 131.68, 131.60, 128.81, 128.71, 127.83, 126.98, 126.46, 126.35, 21.65, 21.62, 21.50, 10.83, 10.81 31 P NMR(162MHz, CDCl3) δ 46.50 HRMS(ESI-TOF):C 15 H 18 Calculated value for OPS [M+H] + :277.0816, Measurement value:277.0818

[0241] 6. Compound 20b [ka] 8b was treated with the organolithium reagent SI-17 on a 1 mmol scale according to general procedure B. Purification by silica gel column chromatography (hexane / EtOAc = 3:1 to 1:1) gave 176 mg (64%, >98:2 er) of the title compound 20b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = -15.1° (c = 0.40 in CHCl3).

[0242] 7. Compound 21a [ka] 8a was treated with the organolithium reagent SI-18 on a 0.2 mmol scale according to general procedure C. Purification by pTLC (hexane / EtOAc = 1:1) gave 47 mg (81%, 61:39 er) of the title compound 21a. Physical state: colorless oil [α] 25 D =-4.9°(c=0.45 in CHCl3) R f =0.45 (hexane / EtOAc = 1:2) 1 H NMR (400MHz, CDCl3) δ 7.82-7.73(m,2H), 7.53-7.39(m,3H), 6.89(dd,J=4.3, 0.6Hz,2H), 2.44(s,6H), 2.39(d,J=11.8Hz,3H), 2.29(s,3H) 13 C NMR (151MHz, CDCl3) δ 143.55, 143.48, 142.30, 142.28, 137.36, 136.66, 131.89, 131.87, 131.39, 131.31, 130 .51, 130.44, 128.92, 128.83, 125.85, 125.16, 23.76, 23.74, 21.19, 21.18, 11.02, 11.00 31 P NMR(162MHz, CDCl3) δ 47.36 HRMS(ESI-TOF):C16 H 20 Calculated value for OPS [M+H] + :291.0972, Measurement value:291.0975

[0243] 8. Compound 21b [ka] 8b was treated with the organolithium reagent SI-18 on a 0.2 mmol scale according to general procedure C. Purification by pTLC (hexane / EtOAc = 1:1) gave 48 mg (83%, 37:63 er) of the title compound 21b. All physical and spectroscopic properties indicated the optical rotation of [α] 25 D = +3.5° (c = 1.00 in CHCl3).

[0244] 9. Compound 22a [ka] 8a and the organolithium reagent SI-27 were treated on a 2 mmol scale according to General Procedure B. Purification by silica gel column chromatography (hexane / EtOAc = 2:1 to 1:2) afforded 364 mg (80%, >98:2 er) of the title compound 22a. Physical state: white solid Melting point = 80-82℃ [α] 25 D =+142.5°(c=1.00 in CHCl3) R f =0.56 (hexane: EtOAc = 1:2) 1 H NMR (400MHz, CDCl3) δ 7.92-7.85(m,2H), 7.56-7.44(m,3H), 2.12(d,J=10.5Hz,3H), 1.17(d,J=16.9Hz,9H) 13C NMR (151MHz, CDCl3) δ 133.23, 133.17, 132.06, 132.04, 130.64, 130.03, 128.47, 128.39, 36.67, 36.20, 24.80, 9.61, 9.59 31 P NMR(162MHz, CDCl3) δ 68.92 HRMS(ESI-TOF):C 11 H 18 Calculated value for OPS [M+H] + :229.0806, Measurement value:229.0818

[0245] 10. Compound 22b [ka] 8b was treated with organolithium reagent SI-27 on a 2 mmol scale according to general procedure B. Purification by silica gel column chromatography (hexane / EtOAc = 2:1 to 1:2) gave 352 mg (77%, >98:2 er) of the title compound 22b. All physical and spectroscopic properties were consistent with the optical rotation of [α]. 25 D = -135.2° (c = 1.00 in CHCl3).

[0246] 11. Compound 23a [ka] 8a and the organolithium reagent SI-19 were treated on a 0.2 mmol scale according to general procedure C. Purification by silica gel column chromatography (hexane / EtOAc = 1:2 to pure EtOAc) gave 15.1 mg (30%, >98:2 er) of the title compound 23a. Physical state: light yellow oil [α] 25 D =-21.6°(c=0.38 in CHCl3) R f =0.34 (Hexane / EtOAc = 1:4) 1H NMR (600MHz, CDCl3) δ 8.81(d,J=4.7Hz,1H), 8.16(ddt,J=7.7, 5.5, 1.1Hz,1H), 8.12-8.02(m,2H), 7.83(tdd,J=7.7, 4.8, 1.7Hz, 1H), 7.57-7.52(m,1H), 7.50-7.45(m,2H), 7.40(dddd,J=7.6, 4.8, 2.7, 1.3Hz,1H), 2.23(d,J=12.2Hz,3H) 13 C NMR (151MHz, CDCl3) δ 156.56, 155.64, 150.74, 150.60, 136.49, 136.42, 132.63, 132.61, 132.18, 132.14, 13 2.07, 131.48, 128.73, 128.66, 128.65, 128.15, 128.00, 125.92, 125.90, 10.21, 10.19 31 P NMR(162MHz, CDCl3) δ 40.29 HRMS(ESI-TOF):C 12 H 13 Calculated value for NOPS [M+H] + :250.0455, Measurement value:250.0459

[0247] 12. Compound 23b [ka] 8b was treated with the organolithium reagent SI-19 on a 0.2 mmol scale according to general procedure C. Purification by silica gel column chromatography (hexane / EtOAc = 1:2 to pure EtOAc) gave 15.9 mg (32%, 98:2 er) of the title compound 23b. All physical and spectroscopic properties indicated the optical rotation was [α]. 25 D = +24.1° (c = 0.80 in CHCl3).

[0248] 13. Compound 24a [ka] 8a was treated with the organolithium reagent SI-23 on a 0.05 mmol scale according to general procedure C. Purification by pTLC (hexane / EtOAc = 1:1) gave 8.6 mg (63%, >98:2 er) of the title compound 24a. Physical state: White amorphous solid [α] 25 D =-10.5°(c=0.43 in CHCl3) R f =0.50 (hexane / EtOAc = 1:2) 1 H NMR (500MHz, CDCl3) δ 8.01(dd,J=15.1, 8.4Hz,2H), 7.66-7.50(m,5H), 7.46(t,J=7.5Hz,1H), 7.38(t,J=8.1Hz,2H), 2.40(d,J=14.1Hz,3H) 13 C NMR (126MHz, CDCl3) δ 133.07, 133.04, 132.93, 132.76, 132.74, 131.90, 131.12, 131.03, 128.98, 1 28.86, 128.75, 119.75, 119.71, 104.97, 104.71, 83.08, 81.64, 11.56, 11.53 31 P NMR(162MHz, CDCl3) δ 18.40 HRMS(ESI-TOF):C 15 H 14 Calculated value for OPS [M+H] + :273.0503, Measurement value:273.0508

[0249] 14. Compound 24b [ka] 8b and the organolithium reagent SI-23 were treated according to general procedure C on a 0.05 mmol scale. Purification by pTLC (hexane / EtOAc = 1:1) gave 8.3 mg (61%, >98:2 er) of the title compound 24b. All physical and spectroscopic properties indicated an optical rotation of [α]25 D = +11.9° (c = 0.42 in CHCl3).

[0250] 15. Compound 25a [ka] 8a was treated with organolithium reagent SI-20 on a 1 mmol scale according to general procedure C. Purification by silica gel column chromatography (hexane / EtOAc = 3:1 to 2:1) afforded 94 mg (32%, 96:4 er) of the title compound 25a. Physical state: White amorphous solid [α] 25 D =-39.2°(c=0.24 in CHCl3) R f =0.44 (Hexane / EtOAc = 1:1) 1 H NMR (400MHz, CDCl3) δ 8.78-8.72(m,1H), 8.08-7.98(m,2H), 7.94-7.84(m,3H), 7.60-7.44(m,6H), 2.34(d,J=12.1Hz,3H) 13 C NMR (151MHz, CDCl3) δ 134.17, 134.10, 133.94, 133.92, 133.67, 133.47, 133.40, 133.26, 133.20, 132.96, 132.53, 132.51, 131.83, 131.7 6, 129.02, 129.01, 128.97, 128.88, 128.82, 128.13, 127.51, 127.12, 127.09, 126.72, 124.58, 124.48, 11.19, 11.17 31 P NMR(162MHz, CDCl3) δ 46.99 HRMS(ESI-TOF):C 17 H 16 Calculated value for OPS [M+H] + :299.0659, Measurement value:299.0662

[0251] 16. Compound 25b [ka] 8b was treated with organolithium reagent SI-20 on a 1 mmol scale according to general procedure C. Purification by silica gel column chromatography (hexane / EtOAc = 3:1 to 2:1) gave 68 mg (23%, 97:3 er) of the title compound 25b. All physical and spectroscopic properties indicated the optical rotation was [α]. 25 D = +37.5° (c = 1.00 in CHCl3).

[0252] c. Introducing a leaving group 1. General operations D [ka] A flame-dried 50 mL round-bottom flask was charged with the starting material (1.0 mmol, 1.0 equiv.). The flask was evacuated and backfilled with argon, and then anhydrous methanol (8 mL) was introduced via syringe, followed by NaOMe (2.0 mL, 0.5 M solution in MeOH, 1.0 mmol, 1.0 equiv.). The resulting mixture was stirred at room temperature for 4 hours, after which saturated aqueous NH4Cl (10 mL) and EtOAc (20 mL) were added. The layers were separated, and the aqueous layer was washed with EtOAc (2 x 10 mL). The organic layers were combined, washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated. (Note: If a white precipitate is observed after concentration, filter again.) The crude product was purified by silica gel chromatography to give the desired product.

[0253] 2. Compound SI-28a [ka] A 1 mmol scale preparation of 11a was carried out according to General Procedure D. Purification by silica gel column chromatography (hexane / EtOAc = 1:1 to 1:4) gave 186 mg (70%, 98:2 er) of the title compound SI-28a. A 16 mmol scale preparation gave 3.24 g (77%, 97:3 er) of SI-28a after purification. Physical state: colorless oil [α] 25 D =-16.5°(c=0.84 in CHCl3) R f =0.25 (hexane / EtOAc = 1:3) 1 H NMR (400MHz, CDCl3) δ 7.98(ddd,J=13.3, 7.5, 1.8Hz,1H), 7.88-7.80(m,2H), 7.50(dddd,J=9.6, 6.8, 2.1, 1.1Hz,2H), 7.46-7.38 (m,2H), 7.06(tdd,J=7.5, 2.6, 0.9Hz,1H), 6.87(dd,J=6.1, 2.2Hz,1H), 3.75(d,J=11.4Hz,3H), 3.71(s,3H) 13 C NMR (151MHz, CDCl3) δ 161.11, 161.08, 134.90, 134.86, 134.59, 134.57, 132.54, 131.93, 131.90, 131.87, 131.86, 1 31.59, 128.18, 128.10, 120.79, 120.71, 119.41, 118.51, 111.39, 111.34, 55.62, 51.53, 51.49 31 P NMR(162MHz, CDCl3) δ 31.78 HRMS(ESI-TOF):C 14 H 16 Calculated value for O3P [M+H] + :263.0837, Measurement value:263.0842

[0254] 3. Compound SI-28b [ka] The organolithium reagent 11b was treated on a 0.9 mmol scale according to general procedure D. Purification by silica gel column chromatography (hexane / EtOAc = 1:1 to 1:4) gave 151 mg (64%, 98:2 er) of the title compound SI-28b. All physical and spectroscopic properties indicated an optical rotation of [α]. 25 D = +19.2° (c = 0.91 in CHCl3), it is identical to SI-28a.

[0255] 4. Compound SI-29a [ka] The organolithium reagent 25a was treated on a 0.30 mmol scale according to general procedure D. Purification by silica gel column chromatography (hexane: / EtOAc = 2:1 to 1:1) afforded 68.4 mg (80%, 92:8 er) of the title compound SI-29a. Physical state: White amorphous solid [α] 25 D =-18.6°(c=0.65 in CHCl3) R f =0.33 (hexane / EtOAc = 1:1) 1 H NMR (600MHz, CDCl3) δ 8.55-8.48(m,1H), 8.21(ddd,J=14.8, 7.1, 1.3Hz,1H), 8.04(d,J=8.1Hz,1H), 7.91-7.85(m,1H), 7.85-7.77(m ,2H), 7.56(ddd,J=8.3, 7.1, 2.8Hz,1H), 7.53-7.47(m,3H), 7.42(td,J=7.5, 3.1Hz,2H), 3.82(d,J=11.1Hz,3H) 13C NMR (151MHz, CDCl3) δ 134.31, 134.26, 133.84, 133.80, 133.78, 133.77, 133.07, 132.99, 132.40, 132.30, 132.28, 131.56, 131.4 9, 129.03, 129.02, 128.73, 128.64, 127.64, 127.21, 126.47, 126.43, 126.33, 124.80, 124.70, 51.75, 51.71 31 P NMR(162MHz, CDCl3) δ 34.77 HRMS(ESI-TOF):C 17 H 16 Calculated value for O2P [M+H] + :283.0888, Measurement value:283.0890

[0256] 5. Compound SI-29b [ka] The organolithium reagent 25b was treated on a 0.22 mmol scale according to General Procedure D. Purification by silica gel column chromatography (hexane / EtOAc = 2:1 to 1:1) gave 50.6 mg (81%, 92:8 er) of the title compound SI-29b. All physical and spectroscopic properties indicated an optical rotation of [α]. 25 D = +14.3° (c = 0.59 in CHCl3), it is identical to SI-29a.

[0257] 6. Compound SI-30a [ka] The organolithium reagent 20a was treated on a 0.66 mmol scale according to general procedure D. Purification by silica gel column chromatography (hexane:EtOAc 2:1 to 1:1) afforded 97 mg (56%, 97:3 er) of the title compound SI-30a. Physical state: colorless oil [α] 25D =+16.1°(c=0.80 in CHCl3) R f =0.30 (hexane / EtOAc = 1:2) 1 H NMR (400MHz, CDCl3) δ 7.83-7.68(m,3H), 7.55-7.48(m,1H), 7.48-7.38(m,2H), 7.10(dt,J=7.9, 2.2H z,1H), 7.03(d,J=4.9Hz,1H), 3.74(d,J=11.1Hz,3H), 2.36(s,3H), 2.34(s,3H) 13 C NMR (126MHz, CDCl3) δ 143.07, 143.05, 142.03, 141.94, 133.66, 133.59, 132.53, 132.50, 132.43, 132.09, 132.07, 131. 72, 131.64, 131.43, 128.63, 128.53, 126.47, 126.36, 125.26, 51.30, 51.25, 21.55, 21.26, 21.22 31 P NMR(162MHz, CDCl3) δ 34.34 HRMS(ESI-TOF):C 15 H 18 Calculated value for O2P [M+H] + :261.1044, Measurement value:261.1050

[0258] 7. Compound SI-30b [ka] The organolithium reagent 20b was treated on a 0.59 mmol scale according to general procedure D. Purification by silica gel column chromatography (hexane:EtOAc 2:1 to 1:1) gave 80 mg (52%, 98:2 er) of the title compound SI-30b. All physical and spectroscopic properties indicated an optical rotation of [α]. 25 D = -16.9° (c = 0.80 in CHCl3).

[0259] d. Substitution 1. General operations E: [ka] A 13x100 mm culture tube, flame-dried under argon, was charged with starting material (0.05 mmol, 1.0 equiv.) in THF (0.5 mL). The Grignard reagent (0.10 mmol, 2.0 equiv., unless otherwise noted) was added dropwise, and the reaction was stirred at room temperature for 12 h. Saturated aqueous NH4Cl (1 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was washed with EtOAc (2x2 mL). The organic layers were combined, washed with brine (1 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by pTLC or silica gel chromatography to give the desired product.

[0260] 2. General operations F: [ka] A flame-dried 13x100 mm culture tube under argon was charged with starting material (0.05 mmol, 1.0 equiv) in THF (0.5 mL) and cooled to -78 °C. An organolithium reagent (0.10 mmol, 2.0 equiv, unless otherwise noted) was added dropwise, and the reaction was stirred at -78 °C for 3 h. Saturated aqueous NH4Cl (1 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was washed with EtOAc (2x2 mL). The organic layers were combined, washed with brine (1 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by pTLC or silica gel chromatography to give the desired product.

[0261] 3. General operations: [ka] Organolithium reagents (0.10 mmol, 2.0 equiv. unless otherwise noted) in 0.3 mL of THF were prepared in a flame-dried 13x100 mm culture tube under argon (see "Synthesis of Grignard and Organolithium Reagents"). Starting material (0.05 mmol, 1.0 equiv.) in THF (0.2 mL) was added dropwise, and the reaction was stirred at -78 °C for 3 h. Saturated aqueous NH4Cl (1 mL) and EtOAc (2 mL) were added. The layers were separated, and the aqueous layer was washed with EtOAc (2x2 mL). The organic layers were combined, washed with brine (1 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by pTLC or silica gel chromatography to give the desired product.

[0262] 4. Compound 26a [ka] SI-28a was treated with the Grignard reagent SI-11 on a 0.05 mmol scale according to general procedure E. Purification by pTLC (5% MeOH in EtOAc) gave 10.9 mg (89%, 98:2 er) of the title compound 26a. A 12 mmol scale afforded 2.54 g (84%, 97:3 er) of 26a after purification. Physical state: white solid Melting point = 81-83℃ [α] 25 D =+30.4°(c=0.55 in CHCl3) R f =0.43(5%MeOH in EtOAc) 1 H NMR (400MHz, CDCl3) δ 7.95(ddd,J=13.1, 7.5, 1.8Hz,1H), 7.79-7.68(m,2H), 7.55-7.37(m,4H), 7.10( t,J=7.5Hz,1H), 6.88(dd,J=8.3, 5.2Hz,1H), 3.72(s,3H), 2.08(d,J=14.0Hz,3H) 13C NMR (151MHz, CDCl3) δ 160.04, 160.01, 135.34, 134.67, 134.06, 134.05, 134.03, 131.42, 131.40, 130.40, 1 30.34, 128.38, 128.30, 121.80, 121.22, 121.15, 111.03, 110.99, 55.42, 16.53, 16.03 31 P NMR (162MHz, CDCl3) δ 29.41 HRMS(ESI-TOF):C 14 H 16 Calculated value for O2P [M+H] + :247.0888, Measurement value:247.0889

[0263] 5. Compound 26b [ka] SI-28b was treated with the Grignard reagent SI-11 on a 0.05 mmol scale according to general procedure E. Purification by pTLC (5% MeOH in EtOAc) gave 10.0 mg (81%, 98:2 er) of the title compound 26b. All physical and spectroscopic properties indicated an optical rotation of [α] 25 D = -28.5° (c = 0.64 in CHCl3).

[0264] 6. Compound 27a [ka] SI-28a was treated with the Grignard reagent SI-12 on a 0.05 mmol scale according to general procedure E. Purification by pTLC (5% MeOH in EtOAc) gave 11.4 mg (87%, 97:3 er) of the title compound 27a.

[0265] 7. Compound 27b [ka] SI-28b was treated with the Grignard reagent SI-12 on a 0.03 mmol scale according to general procedure E. Purification by pTLC (5% MeOH in EtOAc) gave 6.6 mg (85%, 97:3 er) of the title compound 27b. All physical and spectroscopic properties indicated an optical rotation of [α] 25 D = -18.1° (c = 0.33 in CHCl3).

[0266] 8. Compound 28a [ka] SI-28a was treated with the organolithium reagent SI-25 on a 0.05 mmol scale according to general procedure F. Purification by pTLC (hexane / EtOAc = 1:3) gave 10.5 mg (77%, 97:3 er) of the title compound 28a. Physical state: White amorphous solid [α] 25 D =+13.8°(c=0.50 in CHCl3) R f =0.5(EtOH) 1 H NMR (400MHz, CDCl3) δ 8.08(ddd,J=12.4, 7.6, 1.8Hz,1H), 7.88(ddd,J=11.2, 7.9, 1.7Hz,2H), 7.52-7.36(m,4H), 7.09(t,J=7.7Hz,1 H), 6.87(dd,J=8.3, 5.2Hz,1H), 3.83(s,3H), 2.87(dq,J=14.1, 7.1Hz,1H), 1.15(ddd,J=38.7, 17.0, 7.2Hz,6H) 13C NMR (151MHz, CDCl3) δ 159.43, 159.39, 134.88, 134.85, 133.70, 133.65, 133.63, 133.07, 131.26, 131.24, 131.22, 131.20, 128.23 , 128.16, 121.44, 121.37, 121.12, 120.50, 110.64, 110.59, 55.30, 26.66, 26.17, 15.45, 15.44, 15.09, 15.07 31 P NMR(162MHz, CDCl3) δ 38.08 HRMS(ESI-TOF):C 16 H 20 Calculated value for O2P [M+H] + :275.1201, Measurement:275.1203

[0267] 9. Compound 28b [ka] SI-28b was treated with the organolithium reagent SI-25 on a 0.03 mmol scale according to general procedure F. Purification by pTLC (hexane / EtOAc = 1:3) gave 5.6 mg (68%, >98:2 er) of the title compound 28b. All physical and spectroscopic properties indicated an optical rotation of [α] 25 D = +12.6° (c = 0.28 in CHCl3).

[0268] 10. Compound 29a [ka] SI-28a was treated with the organolithium reagent SI-26 on a 0.05 mmol scale according to general procedure F. Purification by pTLC (hexane / EtOAc = 1:3) gave 13.6 mg (94%, 97:3 er) of the title compound 29a. Physical state: colorless oil [α] 25 D =+5.3°(c=0.43 in CHCl3) R f =0.54(EtOH) 1 H NMR (600MHz, acetone-d6) δ 8.01-7.94(m,1H), 7.88-7.80(m,2H), 7.57-7.41(m,4H), 7.14-7.09(m,1H), 7.09-7.04(m,1H), 3.85( s,3H), 2.51-2.40(m,1H), 2.38-2.26(m,1H), 1.61-1.48(m,1H), 1.47-1.35(m,3H), 0.90-0.81(m,3H) 13 C NMR (151 MHz, acetone-d6) δ 160.67, 160.64, 136.89, 136.23, 134.97, 134.94, 134.51, 134.50, 131.82, 131.81, 131.51, 131.45, 128.96, 128.89, 123.05, 122.43, 121.67, 121.60, 111.99, 111.94, 55.72, 29.77, 29.28, 24.64, 24.61, 24.58, 24.54, 13.95 31 P NMR (162 MHz, acetone-d6) δ 29.39 HRMS(ESI-TOF):C 17 H 22 Calculated value for O2P [M+H] + :289.1357, Measurement:289.1362

[0269] 11. Compound 29b [ka]

[0270] SI-28b was treated with the organolithium reagent SI-26 on a 0.03 mmol scale according to general procedure F. Purification by pTLC (hexane / EtOAc = 1:3) gave 7.4 mg (86%, 97:3 er) of the title compound 29b. All physical and spectroscopic properties indicated the optical rotation was [α] 25 D= -7.1° (c = 0.37 in CHCl3).

[0271] 12. Compound 30a [ka] SI-28a was treated with the organolithium reagent SI-27 on a 0.05 mmol scale according to general procedure F. Purification by pTLC (hexane / EtOAc = 1:3) gave 12.0 mg (83%, 90:10 er) of the title compound 30a. Physical state: White amorphous solid [α] 25 D =+10.7°(c=0.27 in CHCl3) R f =0.53(EtOH) 1 H NMR (400MHz, CDCl3) δ 8.16(ddd,J=12.1, 7.6, 1.8Hz,1H), 7.99-7.88(m,2H), 7.54-7.36(m,4H), 7.11(tt, J=7.5, 1.2Hz,1H), 6.91(dd,J=8.3, 5.1Hz,1H), 3.75(s,3H), 1.26(d,J=15.4Hz,9H) 13 C NMR (151MHz, CDCl3) δ 159.73, 159.70, 136.27, 136.24, 133.70, 133.69, 133.27, 132.65, 132.18, 132.12, 131.18, 131.16, 127.97, 127.89, 121.27, 121.20, 120.56, 119.97, 110.97, 110.93, 54.77, 35.10, 34.63, 26.11, 26.10 31 P NMR(162MHz, CDCl3) δ 43.50 HRMS(ESI-TOF):C 17 H 22 Calculated value for O2P [M+H] + :289.1357, Measurement:289.1361

[0272] 13. Compound 30b [ka] SI-28b was treated with the organolithium reagent SI-27 on a 0.03 mmol scale according to general procedure F. Purification by pTLC (hexane / EtOAc = 1:3) gave 6.8 mg (79%, 87:13 er) of the title compound 30b. All physical and spectroscopic properties indicated the optical rotation was [α] 25 D = -12.5° (c = 0.33 in CHCl3).

[0273] 14. Compound 31a [ka] SI-28a was treated with Grignard reagent SI-8 (4.0 equiv.) on a 0.05 mmol scale according to general procedure E. Purification by pTLC (hexane / EtOAc = 1:2) gave 14.5 mg (90%, 96:4 er) of the title compound 31a. Physical state: colorless gel [α] 25 D =-6.7°(c=0.73 in CHCl3) R f =0.35 (hexane / EtOAc = 1:3) 1 H NMR (400MHz, CDCl3) δ 7.75(ddd,J=13.4, 7.6, 1.8Hz,1H), 7.68(ddd,J=12.5, 8.2, 1.3Hz,2H), 7.59(dd,J=12.3, 8.0Hz,2H), 7.55-7.45(m,2H), 7.44-7.37(m,2H), 7.23(dd,J=8.3, 2.7Hz,2H), 7.10-7.02(m,1H), 6.90(dd,J=8.3, 5.2Hz,1H), 3.55(s,3H), 2.38(s,3H) 13C NMR (126MHz, CDCl3) δ 161.01(d,J=3.2Hz), 141.96(d,J=2.6Hz), 135.08(d,J=7.1Hz), 134.28(d,J=1.8 Hz), 133.67(d,J=107.6Hz), 132.01(d,J=10.6Hz), 131.88(d,J=10.2Hz), 131.44( d,J=2.7Hz), 129.85(d,J=109.8Hz), 128.99(d,J=12.9Hz), 128.16(d,J=12.4Hz) , 121.02(d,J=11.6Hz), 120.62(d,J=103.8Hz), 111.50(d,J=6.5Hz), 55.37, 21.71 31 P NMR(162MHz, CDCl3) δ 27.45 HRMS(ESI-TOF):C 20 H 20 Calculated value for O2P [M+H] + :323.1201, Measurement value:323.1200

[0274] 15. Compound 31b [ka] SI-28b was treated with Grignard reagent SI-8 (4.0 equiv.) on a 0.05 mmol scale according to general procedure E. Purification by pTLC (hexane / EtOAc = 1:2) gave 13.6 mg (84%, 97:3 er) of the title compound 31b. All physical and spectroscopic properties indicated an optical rotation of [α] 25 D = +6.5° (c = 0.68 in CHCl3).

[0275] 16. Compound 32a [ka] SI-28a was treated with the Grignard reagent SI-9 on a 0.05 mmol scale according to general procedure E. Purification by pTLC (hexane / EtOAc = 1:3) gave 15.8 mg (93%, 97:3 er) of the title compound 32a. Physical state: White amorphous solid [α] 25 D =-9.5°(c=0.78 in CHCl3) R f =0.33 (hexane / EtOAc = 1:3) 1 H NMR (400MHz, CDCl3) δ 7.76-7.64(m,3H), 7.56-7.46(m,2H), 7.45-7.38(m,2H), 7.36-7.28(m,2H), 7.22(ddt,J=12 .1, 7.5, 1.2Hz,1H), 7.10-7.00(m,2H), 6.91(dd,J=8.2, 5.3Hz,1H), 3.78(s,3H), 3.57(s,3H) 13 C NMR (151 MHz, CDCl3) δ 161.14, 161.12, 159.49, 159.39, 135.10, 135.05, 134.93, 134.44, 134.43, 134.23, 133.61, 132.89, 131.91, 131.85, 131.61, 131.59, 129.46, 129.36, 128.29, 128.20, 124.28, 124.22, 121.08, 121.00, 120.69, 120.01, 117.84, 117.82, 116.77, 116.69, 111.62, 111.57, 55.52, 55.44 31 P NMR(162MHz, CDCl3) δ 27.38 HRMS(ESI-TOF):C 20 H 20 Calculated value for O3P [M+H] + :339.1150, Measurement:339.1150

[0276] 17. Compound 32b [ka] SI-28b was treated with the Grignard reagent SI-9 on a 0.05 mmol scale according to general procedure E. Purification by pTLC (hexane / EtOAc = 1:3) gave 15.8 mg (93%, 97:3 er) of the title compound 32b. All physical and spectroscopic properties indicated the optical rotation was [α] 25 D = +8.1° (c = 0.78 in CHCl3).

[0277] 18. Compound 33a [ka] SI-28a was treated with Grignard reagent SI-7 (10 equiv.) on a 0.05 mmol scale according to general procedure E. Purification by silica gel column chromatography (hexane / EtOAc = 1:2 to pure EtOAc) afforded 16.7 mg (93%, 95:5 er) of the title compound 33a. Physical state: colorless gel [α] 25 D =-7.6°(c=0.50 in CHCl3) R f =0.37 (Hexane / EtOAc = 1:4) 1 H NMR (600MHz, CDCl3) δ 8.37-8.31(m,1H), 7.90-7.83(m,3H), 7.76(dddd,J=23.9, 12.5, 8.0, 1.6Hz,3H), 7.67(ddd,J=10.2, 8.4, 1.5H z,1H), 7.60-7.49(m,4H), 7.44(ddd,J=8.9, 7.0, 3.0Hz,2H), 7.12-7.05(m,1H), 6.96-6.90(m,1H), 3.55(s,3H) 13C NMR (151MHz, CDCl3) δ 161.14, 161.12, 135.19, 135.14, 134.75, 134.73, 134.50, 134.49, 133.79 , 133.73, 133.02, 132.66, 132.57, 131.97, 131.90, 131.66, 131.64, 130.81 , 130.09, 129.09, 128.36, 128.28, 128.03, 127.88, 127.86, 127.78, 127.20, 127.13, 126.76, 121.17, 121.09, 120.71, 120.02, 111.60, 111.56, 55.44 31 P NMR(162MHz, CDCl3) δ 27.55 HRMS(ESI-TOF):C 23 H 20 Calculated value for O2P [M+H] + :359.1201, Measurement:359.1205

[0278] 19. Compound 33b [ka] SI-28b was treated with Grignard reagent SI-7 (10 equiv.) on a 0.05 mmol scale according to general procedure E. Purification by silica gel column chromatography (hexane / EtOAc = 1:2 to pure EtOAc) gave 16.5 mg (92%, 80:20 er) of the title compound 33b. All physical and spectroscopic properties indicated the optical rotation was [α]. 25 D = +8.1° (c = 0.83 in CHCl3).

[0279] 20. Compound 34a [ka] SI-28a was treated with the organolithium reagent SI-20 on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:3) gave 11.7 mg (65%, 98:2 er) of the title compound 34a. Physical state: white crystalline solid Melting point = 167-169℃ [α] 25 D =+21.3(c=0.52 in CHCl3) R f =0.36 (hexane / EtOAc = 1:3) 1 H NMR (400MHz, CDCl3) δ 8.57(d,J=8.5Hz,1H), 7.98(d,J=8.0Hz,1H), 7.87(d,J=8.2Hz,1H), 7.83-7.70(m,3H), 7.5 7-7.36(m,8H), 7.08(tdd,J=7.5, 2.1, 0.9Hz,1H), 6.91(dd,J=7.9, 5.3Hz,1H), 3.46(s,3H) 13 C NMR (151MHz, CDCl3) δ 161.17, 161.15, 134.99, 134.94, 134.36, 134.35, 134.05, 133.94, 133.88, 133 .82, 133.34, 133.18, 133.10, 132.84, 132.82, 132.25, 132.18, 131.58, 131.56 , 129.60, 128.89, 128.80, 128.79, 128.31, 128.23, 127.62, 127.58, 127.17, 126.32, 124.39, 124.29, 121.42, 121.29, 121.22, 120.74, 111.75, 111.71, 55.45 31 P NMR(162MHz, CDCl3) δ 30.93 HRMS(ESI-TOF):C 23 H 20 Calculated value for O2P [M+H] + :359.1201, Measurement:359.1197

[0280] 21. Compound 34b [ka] SI-28b was treated with the organolithium reagent SI-20 on a 0.03 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:3) gave 5.7 mg (53%, 97:3 er) of the title compound 34b. All physical and spectroscopic properties indicated the optical rotation was [α] 25 D = -22.5° (c = 0.29 in CHCl3).

[0281] 22. Compound 35a [ka] SI-28a was treated with the organolithium reagent SI-24 on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:3) gave 19.3 mg (93%, 97:3 er) of the title compound 35a. Physical state: Orange gel [α] 25 D =-38.2°(c=0.10 in CHCl3) R f =0.29 (Hexane / EtOAc = 1:3) 1 H NMR (400MHz, CDCl3) δ 7.95(ddd,J=13.4, 7.5, 1.8Hz,1H), 7.71-7.62(m,2H), 7.54-7.45(m,1H), 7.44(td,J=7.2, 1.6Hz,1H), 7.41-7.34(m,2H), 7. 10(tdd,J=7.5, 1.8, 0.9Hz,1H), 6.87(dd,J=8.3, 5.3Hz,1H), 4.59(s,1H), 4.47(s,1H), 4.44(s,2H), 4.12(s,5H), 3.51(s,3H) 13C NMR (126MHz, CDCl3) δ 160.44, 160.41, 136.23, 135.35, 134.48, 134.43, 133.84, 133.83, 131.00, 130.98, 130.90, 127.90, 127.80, 123.26, 122. 42, 120.92, 120.83, 111.64, 111.59, 73.41, 72.97, 72.87, 72.45, 72.29, 72.19, 71.42, 71.33, 71.17, 71.08, 69.69, 55.35 31 P NMR(162MHz, CDCl3) δ 27.37 HRMS(ESI-TOF):C 23 H 22 Calculated value for FeO2P [M+H] + : 415.0754, Measurement: 415.0747

[0282] 23. Compound 35b [ka] SI-28b was treated with the organolithium reagent SI-24 on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane: EtOAc = 1:3) gave 18.3 mg (88%, 97:3 er) of the title compound 35b. All physical and spectroscopic properties indicated the optical rotation was [α] 25 D = +39.0 (c = 0.10 in CHCl3).

[0283] 24. Compound 36a [ka] SI-28a was treated with the organolithium reagent SI-21 on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:3) gave 18.3 mg (83%, 93:7 er) of the title compound 36a. Physical state: white solid Melting point = 187-189℃ [α] 25 D =+49.6° (c=0.74 in CHCl3) R f =0.25(ヘキサン / EtOAc=1:3) 1 H NMR (400MHz, CDCl3) δ 7.70-7.58(m,3H), 7.55-7.45(m,2H), 7.41-7.30(m,3H), 7.30-7.24(m,2H ), 7.19(dddd,J=7.6,4.2,1.3,0.5Hz,1H),7.03(t,J=8.3Hz,1H),6.89(tdd ,J=7.5,2.0,0.9Hz,1H), 6.76(dd,J=8.0,5.2Hz,1H), 6.28(dd,J=8.4,0.8 Hz,1H), 6.19(dd,J=8.4,0.8Hz,1H), 3.52(s,3H), 3.50(s,3H), 3.43(s,3H) 13 C NMR (126MHz, CDCl3) δ 160.58, 160.55, 157.95, 157.61, 139.20, 139.13, 134.76, 134.70, 134.35, 133.66, 133.56, 133.50, 133.27, 133.26, 132.70, 132.56, 132.48, 132.11, 132.03, 131.01, 1 30.99, 130.70, 130.68, 129.22, 127.51, 127.41, 126.39, 126.28, 122.33, 121.51, 120.60, 120.50, 117.83, 117.80, 111.02, 110.97, 103.04, 102.92, 55.37, 55.28, 55.12 31 P NMR (162MHz, CDCl3) δ 26.90 HRMS (ESI-TOF):C 27 H 26 O4Pとして、calculated value[M+H] + :445.1569, measured value: 445.1565

[0284] 25. Compound 36b

change

[0285] 26. Compound 37a [ka] SI-29a was treated with the organolithium reagent SI-22 (4 equiv.) on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:2) afforded 11.4 mg (56%, 94:6 er) of the title compound 37a. Physical state: yellow amorphous solid [α] 25 D =-23.5°(c=0.38 in CHCl3) R f =0.40 (hexane / EtOAc = 1:2) 1 H NMR (400MHz, acetone-d6) δ 8.65(d,J=8.6Hz,1H), 8.12(d,J=8.2Hz,1H), 8.00(d,J=8.3Hz,1H), 7.79-7.70(m,3H), 7.67(tt,J=7.6, 1.6Hz,1H), 7.64- 7.47(m,7H), 7.45-7.36(m,2H), 3.91(ddd,J=10.1, 8.8, 7.7Hz,1H), 3.80(ddd,J=10.5, 8.4, 7.7Hz,1H), 3.65-3.48(m,2H) 13C NMR (151 MHz, acetone-d6) δ 164.58, 164.56, 135.47, 134.90, 134.84, 134.76, 134.71, 134.64, 134.61, 134.56, 134.15, 134.11, 134.10, 134.02, 133.34, 133.32, 133.11, 133.04, 132.45, 132.4 3, 132.38, 132.36, 131.66, 131.27, 131.21, 130.97, 130.83, 130.75, 129.63, 129.63, 129.15, 129.07, 128.59, 128.56, 127.45, 127.16, 125.25, 125.15, 68.28, 55.74 31 P NMR (162 MHz, acetone-d6) δ 32.05 HRMS(ESI-TOF):C 25 H 21 Calculated value for NO2P [M+H] + :398.1310, Measurement value:398.1306

[0286] 27. Compound 37b [ka] SI-29b was treated with the organolithium reagent SI-22 (4 equiv.) on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:2) gave 8.0 mg (40%, er between 89:11 and 94:6, poor chiral HPLC separation, see below) of the title compound 37b. All physical and spectroscopic properties were consistent with the optical rotation [α] 25 D = +22.8° (c = 0.40 in CHCl3).

[0287] 28. Compound 38a [ka] SI-30a was treated with the organolithium reagent SI-24 on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:3) gave 16.6 mg (84%, 98:2 er) of the title compound 38a. Physical state: Orange amorphous solid [α] 25 D =+88.0°(c=0.10 in CHCl3) R f =0.35 (hexane / EtOAc = 1:3) 1 H NMR (400MHz, CDCl3) δ 7.76-7.67(m,2H), 7.51(dtd,J=16.2, 8.6, 8.1, 6.2Hz,3H), 7.11(dd,J=14.0, 7.8Hz,1H), 6.99(s,1H), 6.9 2(d,J=7.9Hz,1H), 4.72(s,1H), 4.54(s,1H), 4.40(s,1H), 4.21(s,5H), 3.93(s,1H), 2.30(d,J=4.1Hz,6H) 13 C NMR (151MHz, CDCl3) δ 142.33, 142.27, 142.10, 142.08, 134.78, 134.09, 133.58, 133.49, 132.56, 132.49, 131.45, 131.43, 131.32, 131.26, 130.16, 129.45 , 128.33, 128.25, 125.79, 125.71, 74.30, 73.52, 72.84, 72.75, 72.35, 72.28, 71.97, 71.90, 71.33, 71.26, 69.81, 21.47, 21.39, 21.36 31 P NMR(162MHz, CDCl3) δ 31.77 HRMS(ESI-TOF):C 24 H 24 Calculated value for FeOP [M+H] + :413.0961, Measurement value:413.0954

[0288] 29. Compound 38b [ka] SI-30b was treated with the organolithium reagent SI-24 on a 0.05 mmol scale according to general procedure G. Purification by pTLC (hexane / EtOAc = 1:3) gave 15.8 mg (76%, er between 97:3 and 98:2, poor chiral HPLC separation, see below) of the title compound 38b. All physical and spectroscopic properties indicated an optical rotation of [α] 25 D = -79.0° (c = 0.10 in CHCl3).

[0289] e. Stereospecific reduction of phosphine oxides [ka] The reduction of the chiral phosphine was achieved according to a reported procedure (see Rajendran, KV et al., Simple unprecedented conversion of phosphine oxides and sulfides to phosphine boranes using sodium borohydride. Chem. Comm. 2012, 48, 817-819). A solution of 26a (0.5 mmol, 1.0 equiv.) in DCM (1 mL) was added dropwise at room temperature under an argon atmosphere to a stirred solution of Meerwein's salt (0.5 mmol, 1.0 equiv.) in DCM (1 mL). The reaction mixture was gently refluxed for 2 h, at which point the reaction mixture was heated to 100°C. 31 P NMR showed that the conversion of the phosphine oxide to the alkoxyphosphonium salt was complete. After cooling to room temperature, sodium borohydride (3 mmol, 6.0 equiv.) dissolved in diglyme (1 mL) was added dropwise to the reaction mixture. The mixture was gently refluxed for 2 h. 31When P NMR showed complete conversion of the salt to the phosphineborane, the reaction mixture was washed with water (5 mL), and the organic layer was isolated and dried over anhydrous MgSO. The drying agent was removed by filtration, and the solvent was removed under vacuum to give a colorless oil, which was purified by silica gel chromatography (hexane / EtOAc = 4:1) to give 39a (60%, 96:4 er) as a white solid. All spectroscopic data were consistent with literature data. The enantiomeric ratio was determined on a Chiralpak AD-H column using 2% isopropanol in hexane as the eluent at a flow rate of 1 mL / min.

[0290] Example 5 Synthesis of 3'-O-protected nucleosides 1. General Procedure for the Synthesis of 3'-O-Protected Nucleosides (General Procedure G) [ka] The synthesis of 3'-O-TBS-protected nucleosides was adapted from literature procedures. A solution of 5'-O-DMTr-protected nucleoside (10 mmol, 1.0 equiv.) and imidazole (3.14 g, 46.2 mmol, 2.0 equiv.) in DMF (30 mL) was cooled to 0 °C, and TBSCl (20 mmol, 2.0 equiv.) was added. The reaction was stirred at room temperature for 3 days, then diluted with EtOAc (80 mL) and washed with water (100 mL x 3). The organic layer was dried over MgSO4, and the solvent was removed under vacuum. The residue was dissolved in 100 mL of DCM and 40 mL of MeOH and then cooled to 0 °C. A solution of p-toluenesulfonic acid (20 mmol, 2.0 equiv.) in MeOH (10 mL) was added, and the reaction mixture was stirred at 0 °C for 30 min. Pyridine (1 mL) was added to neutralize excess acid. The crude reaction mixture was evaporated to dryness, diluted in EtOAc (100 mL), and washed with saturated NaHCO (50 mL) and brine (50 mL). The organic layer was dried over NaSO, filtered, and concentrated in vacuo. The crude product was purified by silica gel chromatography (gradient from pure DCM to 5% MeOH in DCM) to give the desired product.

[0291] 2. Compound SI-31 [ka] General procedure G was followed on a 10 mmol scale using 5'-O-(4,4'-dimethoxytrityl)thymidine. SI-31 (2.8 g, 79%) was isolated as a white foam with spectroscopic properties identical to those reported in the literature. See Huang, H.-S. et al., A Practical Method for Regioselective 5'-O-tert-Butyldimethylsilyl Deprotection of Persilylated Nucleosides by Methanolic Phosphomolybdic Acid. Synlett 2018, 29, 2437-2443.

[0292] 3. Compound SI-32 [ka] N6-Benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxyadenosine was used on a 10 mmol scale according to General Procedure G. SI-32 (3.8 g, 81%) was isolated as a white foam with spectroscopic properties identical to those reported in the literature. See Molina, AG et al., Acetylated and Methylated β-Cyclodextrins as Viable Soluble Supports for the Synthesis of Short 2'-Oligodeoxyribonucleotides in Solution. Molecules 2012, 17, 12102-12120.

[0293] 4. Compound SI-33 [ka] N4-Benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxycytidine was treated on a 10 mmol scale according to general procedure G. SI-33 (3.7 g, 83%) was isolated as a white foam with spectroscopic properties identical to those reported in the literature. See Huang, H.-S. et al., Synlett 2018, 29, 2437-2443.

[0294] 5. Compound SI-34 [ka] N2-Isobutyryl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxyguanosine was treated on a 10 mmol scale according to general procedure G. SI-34 (3.3 g, 72%) was isolated as a white foam with spectroscopic properties identical to those reported in the literature. See Huang, H.-S. et al., Synlett 2018, 29, 2437-2443.

[0295] Example 6 MPO synthesis a. Loading 1. General operations H [ka] A flame-dried 50 mL round-bottom flask under argon was charged with the nucleoside (1.0 mmol, 1.0 equiv.), 12 (524.6 mg, 2.0 mmol, 2.0 equiv.), and THF (10 mL). DBU (3.0 mmol, 3.0 equiv.) was added dropwise, and the resulting solution was stirred at room temperature for 12 h. The reaction was then quenched with 20 mL of 1x PBS solution and extracted with DCM (3x30 mL). The organic layers were combined, washed with saturated aqueous NaHCO3 (10 mL), brine (10 mL), dried over Na2SO4, filtered, and concentrated. The residue was redissolved in THF (5 mL). Et3N (2 equiv.) and MeI (2 equiv.) were added, and the heterogeneous mixture was stirred vigorously for 1 h. The reaction was quenched with 10 mL of water and extracted with EtOAc (3x20 mL). The organic layers were combined, dried over Na2SO4, filtered and concentrated. The residue was purified by silica gel chromatography to give the desired product.

[0296] Nucleobase Nomenclature: In this disclosure, the following naming conventions will be adopted for nucleobases unless otherwise specified: [ka]

[0297] 2. Compound (R p )-40 [ka] 12b was treated with 5'-O-(4,4'-dimethoxytrityl)thymidine on a 0.5 mmol scale according to general procedure H. Purification by silica gel column chromatography (2% to 5% MeOH in DCM) gave 275 mg (85%, >20:1 dr) of the title compound (R p )-40 was obtained. Physical state: white foam R f =0.3(EtOH) 1H NMR (600MHz, アセトン-d6) δ 10.01(s,1H)、7.58(d,J=1.3Hz,1H)、7.53-7.47(m,2H)、7.41-7.31(m,6H)、7.30 -7.23(m,1H), 6.95-6.89(m,4H), 6.38(dd,J=8.3, 6.0Hz,1H), 5.32(ddt,J=10.9 ,5.3,2.5Hz,1H),4.23(q,J=3.2Hz,1H),3.79(s,6H),3.49-3.40(m,2H),2.63-2 .57(m,2H), 2.18(d,J=13.1Hz,3H), 1.82(d,J=15.7Hz,3H), 1.50(d,J=1.2Hz,3H) 13 C NMR (151MHz, Atotron-d6) δ 164.22, 159.94, 159.93, 151.35, 145.81, 138.60, 136.58, 136.44, 136.24, 13 1.13, 130.15, 129.86, 129.14, 129.13, 128.90, 128.88, 128.39, 127.95, 127.5 1, 126.23, 114.18, 113.71, 111.44, 87.82, 85.26, 85.21, 85.19, 76.47, 76.43, 64.33, 55.67, 55.57, 39.94, 39.92, 21.51, 19.11, 18.39, 12.51, 12.49, 12.28 31 P NMR (162MHz, Atom-d6) δ 56.16 HRMS (ESI-TOF):C 33 H 37 N2O8PSNaとして, calculated value [M+Na] + :675.1906, measured value: 675.1914

[0298] 3. Compound (S) p )-40

change

[0299] 4. Compound (R p )-41 [ka] 12b was treated with N-benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxyadenosine on a 1 mmol scale according to general procedure H. Purification by silica gel column chromatography (2% to 5% MeOH in DCM) gave 628 mg (82%, 20:1 dr) of the title compound (R p )-41 was obtained. Physical state: white foam R f =0.23 (DCM / MeOH=98:2) 1 H NMR (600MHz, acetone-d6) δ 9.97(s,1H), 8.55(s,1H), 8.41(s,1H), 8.12(d,J=7.6Hz,2H), 7.64(t,J=7.4Hz,1H), 7.56(dd,J=8.3, 6. 9Hz,2H), 7.51-7.44(m,2H), 7.38-7.30(m,4H), 7.28(dd,J=8.3, 6.6Hz,2H), 7.25-7.17(m,1H), 6.90-6.8 0(m,4H), 6.60(dd,J=7.7, 6.2Hz,1H), 5.48(ddt,J=8.8, 5.6, 2.7Hz,1H), 4.37(td,J=5.3, 2.5Hz,1H), 3. 77(d,J=1.9Hz,6H), 3.51-3.28(m,3H), 2.92-2.85(m,1H), 2.27(d,J=13.0Hz,3H), 1.85(d,J=15.7Hz,3H) 13C NMR (151 MHz, acetone-d6) δ 165.93, 159.80, 159.78, 153.02, 152.69, 151.42, 146.06, 143.84, 136.77, 136.75, 135.18, 133.34, 131.11, 131.04, 129.88, 129.54, 129.26, 129.16, 129.10, 128.73, 127.75, 126.46, 126.24, 114.04, 87.40, 85.78, 85.73, 76.98, 76.93, 64.29, 55.65, 38.56, 21.53, 19.21, 18.49, 12.65, 12.63 31 P NMR (162 MHz, acetone-d6) δ 55.94 HRMS(ESI-TOF):C 40 H 41 Calculated for N5O7PS [M+H] + :766.2464, Measurement value:766.2474

[0300] 5. Compound (S p )-41 [ka] 12a was treated with N-benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxyadenosine on a 1 mmol scale according to general procedure H. Purification by silica gel column chromatography (2% to 5% MeOH in DCM) afforded 690 mg (90%, >20:1 dr) of the title compound (S p )-41 was obtained. Physical state: white foam R f =0.38 (EtOAc / acetone = 2:1) 1H NMR (600MHz, アセトン-d6) δ 10.06(s,1H), 8.54(s,1H), 8.41(s,1H), 8.11(d,J=7.7Hz,2H), 7.65-7.58(m,1H), 7.53(t,J=7.9Hz,2H), 7.47(dd ,J=8.4,1.3Hz,2H),7.34(d,J=8.6Hz,4H),7.25(t,J=7.7Hz,2H),7.21-7.16(m,1H),6.82(t,J=9.1Hz,4H),6.59(d d,J=7.8,6.2Hz,1H),5.49(ddd,J=12.3,5.5,2.5Hz,1H),4.52(td,J=5.0,2.5Hz,1H),3.75(d,J=2.6Hz,6H),3.51- 3.41(m,2H), 3.41-3.33(m,1H), 2.77(ddd,J=14.1,6.2,2.7Hz,1H), 2.33(d,J=13.0Hz,3H), 1.85(d,J=15.7Hz,3H) 13 C NMR (151MHz, Atotron-d6) δ 165.90, 159.57, 159.55, 152.86, 152.55, 151.28, 145.98, 143.66, 136.67, 135.01, 133.17, 130.99, 130.93, 129.36, 129.15, 128.99, 128.55, 127.53, 126.26, 113.87, 87.18, 86.17, 86.14, 85.63, 76.94, 76.89, 64.25, 55.50, 38.10, 38.07, 19.21, 18.49, 12.35, 12.33 31 P NMR (162MHz, Atom-d6) δ 56.03 HRMS (ESI-TOF):C 40 H 41 N5O7PSとして, calculated value [M+H] + :766.2464, measured value:766.2480

[0301] 6. Compound (R) p )-42

change

[0302] 7. Compound (S p )-42 [ka] 12a was treated with N-isobutyryl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxyguanosine on a 1 mmol scale according to general procedure H. Purification by silica gel column chromatography (2% to 5% MeOH in DCM) gave 441 mg (59%, >20:1 dr) of the title compound (S p )-42 was obtained. Physical state: white foam R f =0.25 (EtOAc / acetone = 2:1) 1 H NMR (600MHz, acetone-d6) δ 12.04(s,1H), 10.84(s,1H), 7.92(s,1H), 7.41-7.37(m,2H), 7.30-7.15(m,7H), 6.79(dd,J=9.0, 2.5H z,4H), 6.40(dd,J=8.0, 5.8Hz,1H), 5.59(ddt,J=10.8, 5.6, 2.7Hz,1H), 4.62-4.55(m,1H), 3.75(d,J= 2.1Hz,6H), 3.43(dd,J=10.4, 5.0Hz,1H), 3.32(dd,J=10.4, 3.8Hz,1H), 3.06(ddd,J=13.9, 8.0, 5.9Hz ,1H), 2.82-2.72(m,2H), 2.38(d,J=13.4Hz,3H), 1.98(d,J=15.6Hz,3H), 1.15(dd,J=34.6, 6.8Hz,6H) 13C NMR (151 MHz, acetone-d6) δ 180.81, 159.55, 155.79, 149.27, 149.24, 145.80, 137.82, 136.54, 130.88, 130.86, 128.95, 128.54, 127.56, 122.03, 113.83, 87.07, 85.87, 85.85, 84.46, 76.90, 76.85, 64.11, 55.47, 39.14, 39.11, 36.44, 19.35, 19.31, 19.19, 18.60, 12.46, 12.43 31 P NMR (162 MHz, acetone-d6) δ 58.75 HRMS(ESI-TOF):C 37 H 43 Calculated for N5O8PS [M+H] + :748.2570, Measurement value:748.2573

[0303] 8. Compound (R p )-43 [ka] 12b was treated with N4-benzoyl-5'-O-(4,4'-dimethoxytrityl)-2'-deoxycytidine on a 1 mmol scale according to general procedure H. Purification by silica gel column chromatography (2% to 5% MeOH in DCM) gave 652 mg (88%, >20:1 dr) of the title compound (R p )-43 was obtained. Physical state: white foam R f =0.21 (DCM / MeOH=98:2) 1H NMR (600MHz, アセトン-d6) δ 9.79(s,1H), 8.25(d,J=7.4Hz,1H), 8.17(ddt,J=7.5,6.2,2.6Hz,2H), 7.68-7.65(m,1H), 7.61-7.55(m,2 H), 7.51(dt,J=8.8,2.2Hz,2H), 7.42-7.34(m,5H), 7.31-7.17(m,3H), 6.96-6.92(m,4H), 6.29(t,J=6.4Hz ,1H), 5.27(ddt,J=10.4,7.2,3.8Hz,1H), 4.38(q,J=3.8Hz,1H), 3.82(s,6H), 3.54(d,J=4.0Hz,2H), 2.88( ddd,J=14.4,6.3,3.5Hz,1H), 2.50(dt,J=12.3,5.9Hz,1H), 2.22(d,J=13.0Hz,3H), 1.84(d,J=15.7Hz,3H) 13 C NMR (151MHz, Atotron-d6) δ 159.94, 145.68, 145.16, 138.63, 136.51, 136.45, 133.75, 131.18, 131.16, 129.89, 129.61, 129.24, 129.21, 129.17, 128.90 , 127.96, 126.25, 114.20, 87.89, 87.76, 85.94, 85.89, 75.80, 75.76, 63.74, 55.67, 41.31, 21.54, 19.21, 18.48, 12.53, 12.51 31 P NMR (162MHz, Atom-d6) δ 56.15 HRMS (ESI-TOF):C 39 H 41 N3O8PSとして, calculated value [M+H] + :742.2352, measured value:742.2360

[0304] 9. Compound (S) p )-43

change

[0305] b. Coupling 1. General operations I [ka] The coupling step was performed according to a modified version of the reported procedure. A culture tube was charged with LiCl (12.7 mg, 0.3 mmol, 10 equiv.), flame-dried under vacuum, and allowed to cool to room temperature. The loaded compound (0.06 mmol, 2.0 equiv.) and 3'-TBS-nucleoside (0.03 mmol, 1.0 equiv.) were added to the culture tube, which was then sealed and backfilled with Ar three times. DMF (0.5 mL) was added, and the reaction was stirred for approximately 15 min until all LiCl was dissolved. DBU (91.3 mg, 0.09 mL, 0.6 mmol, 20 equiv.) was then introduced via syringe, and the resulting mixture was stirred overnight for 16 h. The reaction was quenched by adding 1x PBS solution (10 mL) and extracted with DCM (3x10 mL). The organic layers were combined, washed with water (2x10 mL) and brine (10 mL), dried over MgSO4, filtered, and concentrated. If necessary, the remaining DMF can be removed azeotropically with toluene. The residue was purified by pTLC to give the desired product.

[0306] 2. Compound (S p )-44 [ka] (S p )-40 and SI-31 were treated on a 0.03 mmol scale according to General Procedure I. Purification by pTLC (EtOAc / DCM:acetone=5:5:2) afforded 23.9 mg (83%, >20:1 dr) of the title compound (S p )-44 was obtained. Physical state: White amorphous solid Rf =0.32 (EtOAc / DCM / acetone = 5:5:2) 1 H NMR (600MHz, acetone-d6) δ 10.12(s,1H), 10.06(s,1H), 7.58(q,J=1.2Hz,1H), 7.54(q,J=1.3Hz,1H), 7.51-7.47(m,2H), 7.39-7.31(m,6H), 7.28-7.23(m,1H), 6.93 -6.88(m,4H), 6.37(t,J=7.1Hz,1H), 6.30(dd,J=7.7, 6.1Hz,1H), 5.34(dtd,J=8.3, 4.2, 2.6Hz,1H), 4.52(dt,J=6.1, 3.2Hz,1H), 4.27(q ,J=3.0Hz,1H), 4.26-4.22(m,1H), 4.09(ddd,J=11.4, 7.0, 3.5Hz,1H), 4.00(dt,J=5.2, 3.3Hz,1H), 3.79(s,6H), 3.48-3.39(m,2H), 2.61 -2.55(m,2H), 2.31-2.16(m,2H), 1.81(d,J=1.3Hz,3H), 1.59(d,J=17.7Hz,3H), 1.45(d,J=1.2Hz,3H), 0.91(s,9H), 0.12(d,J=2.0Hz,6H) 13 C NMR (151 MHz, acetone-d6) δ 164.26, 164.19, 159.80, 159.78, 151.29, 151.26, 145.73, 136.55, 136.45, 136.33, 136.17, 131.05, 131.04, 129.05, 128.79, 127.83, 114.07, 111.30, 111.11, 87.74, 86 .32, 86.27, 85.74, 85.49, 85.46, 85.07, 76.99, 76.95, 73.06, 65.66, 65.62, 64.22, 55.57, 40.73, 39.57, 39.54, 26.13, 18.49, 12.57, 12.18, 12.13, 11.24, -4.56, -4.65 31 P NMR (162 MHz, acetone-d6) δ 33.33 HRMS(ESI-TOF):C 48 H 61 N4O 13Calculated value as PSi [M+Na] + :983.3640, Measurement value:983.3629

[0307] 3. Compound (S p )-45 [ka] (S p )-40 and SI-32 were treated on a 0.03 mmol scale according to General Procedure I. Purification by pTLC (EtOAc / DCM / acetone = 5:5:2) afforded 29.3 mg (91%, 20:1 dr) of the title compound (S p )-45 was obtained. Physical state: White amorphous solid R f =0.32 (EtOAc / DC / acetone = 5:5:2) 1 H NMR (600MHz, acetone-d6) δ 10.25(s,1H), 10.06(s,1H), 8.64(s,1H), 8.46(s,1H), 8.10(d,J=7.7Hz,2H), 7.64-7.60(m,1H), 7.57-7.51(m,3H), 7.47- 7.44(m,2H), 7.36-7.28(m,6H), 7.24-7.19(m,1H), 6.88(dd,J=9.0, 1.2Hz,4H), 6.56(t,J=6.5Hz,1H), 6.33(dd,J=8.0, 6. 1Hz,1H), 5.26(ddt,J=8.2, 5.5, 2.7Hz,1H), 4.88(dt,J=6.0, 4.0Hz,1H), 4.26-4.10(m,4H), 3.76(s,6H), 3.41-3.31(m,2H) ), 3.06(dt,J=13.0, 6.2Hz,1H), 2.56-2.49(m,3H), 1.50(d,J=17.7Hz,3H), 1.42(d,J=1.2Hz,3H), 0.95(s,9H), 0.18(s,6H) 13C NMR (151 MHz, acetone-d6) δ 165.88, 164.25, 159.75, 159.73, 152.85, 152.63, 151.33, 151.23, 145.74, 143.57, 136.45, 136.31, 136.22, 135.01, 133.18, 131.03, 131.01, 129.37, 129.21, 129.01, 128.77, 127.77, 128.77 6.04, 114.05, 111.26, 87.69, 86.64, 86.60, 85.43, 85.40, 85.36, 85.09, 76.91, 76.87, 73.07, 65.14, 65.10, 64.14, 55.55, 40.44, 39.57, 39.55, 26.18, 18.55, 12.11, 11.16, -4.51, -4.61 31 P NMR (162 MHz, acetone-d6) δ 32.99 HRMS(ESI-TOF):C 55 H 64 N7O 12 Calculated value as PSi [M+H] + :1074.4198, Measurement value:1074.4181

[0308] 4. Compound (S p )-46 [ka] (S p )-40 and SI-33 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (15:1 EtOAc / acetone) gave 25.5 mg (81%, >20:1 dr) of the title compound (S p )-46 was obtained. Physical state: White amorphous solid R f =0.50 (EtOAc / acetone = 10:1) 1H NMR(600MHz、アセトン-d6) δ 10.09(s,1H)、9.95(s,1H)、8.22(d,J=7.6Hz,1H)、8.17-8.13(m,2H)、7.67-7.62(m,1H)、7.58(d,J=1.3Hz,1H)、7.55(t,J=7.8Hz,2H)、7.50-7.46(m,2H)、7.39-7.29(m,7H)、7.26-7.21(m,1H)、6.89(d,J=9.0Hz,4H)、6.37(t,J=7.0Hz,1H)、6.23(t,J=6.3Hz,1H)、5.35(dtd,J=8.1、4.3、2.5Hz,1H)、4.54(dt,J=6.1、4.0Hz,1H)、4.35-4.28(m,2H)、4.17(ddd,J=11.4、6.8、3.5Hz,1H)、4.12(q,J=4.1Hz,1H)、3.77(s,6H)、3.45(ddd,J=31.5、10.6、3.3Hz,2H)、2.60(dd,J=7.1、4.4Hz,2H)、2.46(ddd,J=13.6、6.3、4.1Hz,1H)、2.24(dt,J=13.5、6.3Hz,1H)、1.61(d,J=17.7Hz,3H)、1.45(d,J=1.3Hz,3H)、0.91(s,9H)、0.12(d,J=1.6Hz,6H) 13 C NMR(151MHz、アセトン-d6) δ 164.19、163.70、159.77、159.76、151.27、145.73、145.27、136.46、136.32、136.25、134.62、133.59、131.04、129.44、129.16、129.04、128.78、127.81、114.06、114.05、111.24、97.15、87.85、87.72、86.91、86.87、85.48、85.45、85.20、77.05、77.01、72.59、65.27、65.23、64.24、55.55、42.10、39.57、39.55、26.14、18.49、12.22、12.15、11.28、-4.53、-4.68 31 P NMR(162MHz、アセトン-d6) δ 33.43 HRMS(ESI-TOF):C 54 H64 N5O 13 Calculated value as PSi [M+H] + :1050.4086, Measurement value:1050.4083

[0309] 5. Compound (S p )-47 [ka] (S p )-40 and SI-34 were treated according to General Procedure I on a 0.03 mmol scale. Purification by pTLC (10:1 EtOAc / acetone) afforded 25.0 mg (80%, >20:1 dr) of the title compound (S p )-47 was obtained. Physical state: White amorphous solid R f =0.37 (EtOAc / acetone = 10:1) 1H NMR(600MHz、ァ-d6) δ 12.06(s,1H), 11.00(s,1H), 10.06(s,1H), 7.97(s,1H), 7.58(d,J=1.3Hz,1H) 7.48(dd,J=8.4; 1.2Hz,2H) 7.38-7.29(m,6H) 7.26-7.20(m,1H) 6.91-6.86( m,4H) 6.38(dd,J=7.7;6.5Hz,1H) 6.33(dd,J=8.9;5.7Hz,1H) 5.36(ddt,J=7 .8、5.8、3.1Hz,1H)、4.63(dt,J=5.2、1.8Hz,1H)、4.45(ddd,J=11.0、7.0、4.9Hz ,1H)、4.37-4.29(m,2H)、4.18(td,J=5.4、5.0、1.6Hz,1H)、3.77(d,J=1.7Hz,6H). )、3.45(qd,J=10.6、3.4Hz,2H)、2.99-2.89(m,1H)、2.84-2.80(m,1H)、2.63-2 58(m,2H)、2.27(ddd,J=13.3、5.8、1.9Hz,1H)、1.62(d,J=17.6Hz,3H)、1.45(d, J=1.1Hz,3H) 1.16(dd,J=16.4;6.9Hz,6H) 0.92(s,9H) 0.13(d,J=3.7Hz,6H) 13 C NMR(151MHz、ァ-d6) δ 181.00, 164.13, 159.80, 159.78, 155.81, 151.25, 149.07, 148.92, 145.67, 139.55, 136.40 136.30, 136.22, 131.02, 131.00, 129.04, 128.78, 127.85, 123.14, 114.05, 111.28, 87.74, 7.07, 87.02, 86.90, 85.48, 85.45, 85.25, 77.51, 77.48, 73.92, 66.07, 66.03, 64.26, 55.56 39.92, 39.47, 39.44, 36.41, 26.17, 19.40, 19.23, 18.56, 12.14, 11.71, 10.78, -4.54, -4.61 31 P NMR(162MHz。。ツ-d6) δ 34.75 HRMS(ESI-TOF):C52 H 66 N7O 13 Calculated value as PSi [M+Na] + :1078.4123, Measurement value:1078.4097

[0310] 6. Compound (S p )-48 [ka] (S p )-41 and SI-31 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (EtOAc / acetone = 7:1) afforded 24.2 mg (75%, >20:1 dr) of the title compound (S p )-48 was obtained. Physical state: White amorphous solid R f =0.32 (EtOAc / acetone = 7:1) 1H NMR(600MHz、アセトン-d6) δ 10.28(s,1H)、10.08(s,1H)、8.55(s,1H)、8.41(s,1H)、8.12(d,J=7.6Hz,2H)、7.63(t,J=7.4Hz,1H)、7.59(d,J=1.3Hz,1H)、7.54(t,J=7.8Hz,2H)、7.45(dd,J=8.5、1.3Hz,2H)、7.35-7.30(m,4H)、7.26(t,J=7.6Hz,2H)、7.22-7.16(m,1H)、6.86-6.81(m,4H)、6.58(dd,J=7.7、6.2Hz,1H)、6.34(dd,J=7.7、6.1Hz,1H)、5.47(td,J=5.7、2.9Hz,1H)、4.56(dt,J=6.1、3.2Hz,1H)、4.42(td,J=5.1、2.6Hz,1H)、4.32(ddd,J=11.4、7.7、5.2Hz,1H)、4.19(ddd,J=11.4、7.1、3.5Hz,1H)、4.05(dt,J=5.5、3.2Hz,1H)、3.76(d,J=2.6Hz,6H)、3.51-3.41(m,2H)、3.37(ddd,J=13.9、7.8、6.1Hz,1H)、2.81(ddd,J=14.1、6.2、2.8Hz,1H)、2.34-2.21(m,2H)、1.83(d,J=1.2Hz,3H)、1.63(d,J=17.7Hz,3H)、0.91(s,9H)、0.13(s,6H) 13 C NMR(151MHz、アセトン-d6) δ 164.56、163.02、158.28、158.26、151.54、151.22、150.01、144.58、142.37、135.30、135.26、133.68、131.86、129.65、129.58、128.03、127.87、127.66、127.26、126.27、124.90、112.56、109.82、85.93、85.05、85.01、84.61、84.58、84.42、84.27、75.95、75.91、71.72、64.36、64.32、62.93、54.18、39.46、36.94、36.91、24.81、17.16、11.27、10.92、9.97、-5.88、-5.96 31P NMR (162 MHz, acetone-d6) δ 33.03 HRMS(ESI-TOF):C 55 H 64 N7O 12 Calculated value as PSi [M+H] + :1074.4198, Measurement value:1074.4185

[0311] 7. Compound (S p )-49 [ka] (S p )-41 and SI-32 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (10% MeOH in EtOAc) gave 27.5 mg (77%, >20:1 dr) of the title compound (S p )-49 was obtained. Physical state: White amorphous solid R f =0.28(10%MeOH in EtOAc) 1H NMR(600MHz、アセトン-d6) δ 9.99(d,J=12.5Hz,2H)、8.66(s,1H)、8.52(d,J=7.4Hz,2H)、8.42(s,1H)、8.13(t,J=6.2Hz,4H)、7.65(dt,J=15.2、7.4Hz,2H)、7.56(dt,J=15.6、7.6Hz,4H)、7.47-7.42(m,2H)、7.34-7.30(m,4H)、7.26(dd,J=8.5、7.0Hz,2H)、7.21-7.16(m,1H)、6.85-6.80(m,4H)、6.61(t,J=6.5Hz,1H)、6.55(dd,J=7.7、6.2Hz,1H)、5.40(ddt,J=8.4、5.8、2.7Hz,1H)、4.93(dt,J=6.0、4.0Hz,1H)、4.35(td,J=5.1、2.5Hz,1H)、4.31(dd,J=7.2、4.4Hz,2H)、4.20(q,J=4.2Hz,1H)、3.76(d,J=2.0Hz,6H)、3.46-3.36(m,2H)、3.31(dt,J=14.2、7.6Hz,1H)、3.09(dt,J=12.9、6.2Hz,1H)、2.76(ddd,J=14.0、6.2、2.8Hz,1H)、2.58(ddd,J=13.4、6.5、4.3Hz,1H)、1.55(d,J=17.7Hz,3H)、0.97(s,9H)、0.21(s,6H) 13 C NMR(151MHz、アセトン-d6) δ 165.92、165.85、159.60、159.58、152.91、152.82、152.65、152.51、151.20、145.95、143.83、143.55、136.66、136.65、135.07、134.99、133.22、133.19、130.98、130.92、129.41、129.39、129.20、129.17、128.99、128.58、127.56、126.48、126.29、126.19、113.89、87.21、86.72、86.68、85.88、85.85、85.64、85.42、77.27、77.24、73.09、65.14、65.10、64.24、55.51、40.64、38.14、26.20、18.57、12.11、11.16、-4.50、-4.58 31 P NMR (162 MHz, acetone-d6) δ 32.60 HRMS(ESI-TOF):C 62 H 67 N 10 O 11 Calculated value as PSi [M+H] + :1187.4576, Measurement value:1187.4561

[0312] 8. Compound (S p )-50 [ka] (S p )-41 and SI-33 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (EtOAc / acetone = 7:1) afforded 26.8 mg (77%, >20:1 dr) of the title compound (S p )-50 was obtained. Physical state: White amorphous solid R f =0.50 (EtOAc / acetone = 5:1) 1H NMR(600MHz、アセトン-d6) δ 10.06(s,2H)、8.55(s,1H)、8.46(s,1H)、8.31(d,J=7.5Hz,1H)、8.16-8.10(m,4H)、7.67-7.60(m,2H)、7.55(q,J=8.6、7.7Hz,4H)、7.47(d,J=7.3Hz,2H)、7.41(s,1H)、7.35-7.31(m,4H)、7.26(t,J=7.7Hz,2H)、7.22-7.17(m,1H)、6.86-6.80(m,4H)、6.62(dd,J=7.6、6.2Hz,1H)、6.28(t,J=6.3Hz,1H)、5.50(ddt,J=8.5、5.8、2.6Hz,1H)、4.60(dt,J=6.2、4.0Hz,1H)、4.46(dt,J=8.0、3.9Hz,1H)、4.40(ddd,J=11.8、7.3、4.8Hz,1H)、4.31(ddt,J=10.3、6.9、3.3Hz,1H)、4.18(q,J=4.0Hz,1H)、3.76(dd,J=2.4、1.5Hz,6H)、3.53-3.44(m,2H)、3.41(dt,J=13.7、6.7Hz,1H)、2.85(ddd,J=14.0、6.2、2.8Hz,1H)、2.50(ddt,J=11.8、5.7、2.8Hz,1H)、2.29(dt,J=13.2、6.3Hz,1H)、1.67(d,J=17.7Hz,3H)、0.93(s,9H)、0.15(s,6H) 13 C NMR(151MHz、アセトン-d6) δ 166.11、163.70、159.60、159.58、152.81、152.56、151.17、145.92、145.34、143.93、136.64、136.61、134.96、134.56、133.60、133.27、130.98、130.91、129.44、129.39、129.18、129.14、128.99、128.58、127.59、126.14、113.89、97.10、87.94、87.24、86.98、86.94、85.97、85.94、85.70、77.37、77.33、72.57、65.30、65.26、64.29、55.51、42.21、38.24、38.22、26.15、18.49、12.31、11.36、-4.52、-4.65 31 P NMR (162 MHz, acetone-d6) δ 33.17 HRMS(ESI-TOF):C 61 H 67 N8O 12 Calculated value as PSi [M+H] + :1163.4464, Measurement value:1163.4448

[0313] 9. Compound (S p )-51 [ka] (S p )-41 and SI-34 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (10% MeOH in EtOAc) gave 20.0 mg (60%, >20:1 dr) of the title compound (S p )-51 was obtained. Physical state: White amorphous solid R f =0.25 (EtOAc:acetone = 5:1) 1H NMR(600MHz、アセトン-d6) δ 12.08(s,1H)、11.07(s,1H)、10.00(s,1H)、8.54(s,1H)、8.43(s,1H)、8.13(d,J=7.6Hz,2H)、8.01(s,1H)、7.67-7.61(m,1H)、7.55(t,J=7.7Hz,2H)、7.47-7.40(m,2H)、7.34-7.27(m,4H)、7.23(dd,J=8.3、6.8Hz,2H)、7.20-7.15(m,1H)、6.80(t,J=8.7Hz,4H)、6.62(dd,J=7.6、6.2Hz,1H)、6.36(dd,J=8.8、5.7Hz,1H)、5.50(ddt,J=8.4、5.8、2.7Hz,1H)、4.71-4.64(m,1H)、4.53(ddd,J=11.6、7.1、4.7Hz,1H)、4.47(td,J=5.1、2.5Hz,1H)、4.40(dt,J=11.1、6.1Hz,1H)、4.22(t,J=4.5Hz,1H)、3.74(d,J=2.7Hz,6H)、3.50-3.38(m,3H)、2.99(ddd,J=13.7、8.8、5.2Hz,1H)、2.85-2.82(m,2H)、2.31(ddd,J=13.2、5.8、1.9Hz,1H)、1.65(d,J=17.6Hz,3H)、1.15(dd,J=24.8、6.8Hz,6H)、0.93(s,9H)、0.15(d,J=1.4Hz,6H) 13C NMR (151 MHz, acetone-d6) δ 181.01, 165.88, 159.61, 159.58, 155.87, 152.84, 152.51, 151.32, 149.11, 148.97, 145.85, 143.87, 139.57, 136.61, 136.57, 135.03, 133.23, 130.95, 130.88, 129.41, 129.17, 128.99, 128.56, 127.61, 126. 31, 123.15, 113.86, 87.20, 87.14, 87.10, 86.92, 86.00, 85.97, 85.71, 77.72, 77.68, 73.91, 66.12, 66.08, 64.35, 55.51, 40.06, 38.10, 38.07, 36.41, 26.18, 19.39, 19.23, 18.56, 11.84, 10.91, -4.50, -4.57 31 P NMR (162 MHz, acetone-d6) δ 34.58 HRMS(ESI-TOF):C 59 H 69 N 10 O 12 Calculated value as PSi [M+H] + :1169.4682, Measurement value:1169.4675

[0314] 10. Compound (S p )-52 [ka] (S p )-43 and SI-31 were treated on a 0.03 mmol scale according to General Procedure I. Purification by pTLC (EtOAc / DCM:acetone=5:5:3) afforded 14.2 mg (46%, >20:1 dl) of the title compound (S p )-52 was obtained. Physical state: White amorphous solid R f =0.33 (EtOAc / DCM:acetone=5:5:3) 1H NMR(600MHz、アセトン-d6) δ 9.98(s,1H)、9.90(s,1H)、8.24(d,J=7.5Hz,1H)、8.18-8.12(m,2H)、7.68-7.63(m,1H)、7.58-7.53(m,3H)、7.50-7.47(m,2H)、7.39-7.32(m,6H)、7.28-7.19(m,2H)、6.95-6.88(m,4H)、6.31(dd,J=7.7、6.1Hz,1H)、6.23(t,J=6.2Hz,1H)、5.27(ddt,J=7.7、6.2、3.9Hz,1H)、4.54(dt,J=6.1、3.1Hz,1H)、4.37(q,J=3.8Hz,1H)、4.26(ddd,J=11.4、7.7、5.2Hz,1H)、4.10(ddd,J=11.4、7.0、3.6Hz,1H)、4.02(dt,J=5.2、3.3Hz,1H)、3.80(d,J=1.5Hz,6H)、3.55-3.46(m,2H)、2.82-2.77(m,1H)、2.52(dt,J=14.2、6.3Hz,1H)、2.31-2.19(m,2H)、1.82(d,J=1.2Hz,3H)、1.61(d,J=17.7Hz,3H)、0.91(s,9H)、0.12(d,J=1.8Hz,6H) 13 C NMR(151MHz、アセトン-d6) δ 164.20、163.77、159.77、151.25、145.56、145.05、136.53、136.40、136.34、134.60、133.62、131.05、131.02、129.47、129.13、129.09、128.80、127.81、114.09、111.10、97.08、87.79、87.60、86.33、86.29、86.00、85.97、85.76、75.89、75.85、73.08、65.57、65.53、63.41、55.55、40.85、40.83、40.75、26.14、18.50、12.60、12.20、11.25、-4.55、-4.64 31 P NMR(162MHz、アセトン-d6) δ 33.25 HRMS(ESI-TOF):C 54 H 64 N5O 13Calculated value as PSi [M+Na] + :1072.3905, Measurement value:1072.3926

[0315] 11. Compound (S p )-53 [ka] (S p )-43 and SI-32 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (10:10:7 EtOAc / DCM / acetone) afforded 27.6 mg (79%, >20:1 dr) of the title compound (S p )-53 was obtained. Physical state: White amorphous solid R f =0.34 (EtOAc / DCM / acetone = 5:5:3) 1 H NMR (600MHz, acetone-d6) δ 10.03(s,2H), 8.64(s,1H), 8.47(s,1H), 8.22(d,J=7.5Hz,1H), 8.13(ddd,J=8.7, 4.0, 2.6Hz,4H), 7.65-7.59(m,2H), 7.56-7.50(m,4H), 7.46-7.41(m,2H), 7.34-7.28(m,6H), 7.27-7.15(m,2H), 6.90-6.85(m,4H), 6.56(t,J=6.5Hz,1H), 6.18(t,J=6.1Hz,1H), 5.21(ddt,J=8. 0, 6.3, 4.2Hz,1H), 4.90(dt,J=5.9, 4.0Hz,1H), 4.28-4.17(m,3H), 4.14(q,J=4.3Hz,1H), 3.77(d,J=1.7Hz,6H), 3.42(dd,J=3.6, 1.2Hz,2 H), 3.06(dt,J=13.5, 6.2Hz,1H), 2.73(ddd,J=14.1, 6.3, 4.4Hz,1H), 2.57-2.46(m,2H), 1.55(d,J=17.8Hz,3H), 0.95(s,9H), 0.17(s,6H) 13C NMR (151 MHz, acetone-d6) δ 165.90, 163.82, 159.69, 152.86, 152.61, 151.24, 145.50, 145.05, 143.58, 136.40, 136.29, 135.06, 134.59, 133.55, 133.16, 130.99, 130.95, 129.39, 129.22, 129.19, 129.08, 129.04, 128.76, 1 27.72, 126.18, 114.06, 97.01, 87.71, 87.53, 86.63, 86.59, 85.77, 85.74, 85.34, 75.55, 75.51, 73.10, 65.04, 65.00, 63.17, 55.53, 40.73, 40.71, 40.45, 26.19, 18.56, 12.15, 11.21, -4.51, -4.60 31 P NMR (162 MHz, acetone-d6) δ 33.13 HRMS(ESI-TOF):C 61 H 67 N8O 12 Calculated value as PSi [M+H] + :1163.4464, Measurement value:1163.4436

[0316] 12. Compound (S p )-54 [ka] (S p )-43 and SI-33 were treated on a 0.03 mmol scale according to General Procedure I. Purification by pTLC (DCM / acetone = 2:1) afforded 22.4 mg (66%, >20:1 dr) of the title compound (S p )-54 was obtained. Physical state: White amorphous solid R f =0.31 (DCM / acetone = 2:1) 1H NMR(600MHz、ァ-d6) δ 10.02(s,2H) 8.24(dd,J=24.8;7.5Hz,2H)8.15(ddd,J=8.6;2.4;1.3Hz,4H)7. 65-7.61(m,2H) 7.53(dd,J=8.2; 7.4Hz,4H) 7.47(dd,J=8.5; 1.3Hz,2H) 7.38-7. 31(m,7H)、7.25-7.20(m,2H)、6.89(dd,J=9.0、1.9Hz,4H)、6.24(td,J=6.3、1.8Hz, 2H)、5.29(ddt,J=8.0、6.3、4.2Hz,1H)、4.57(dt,J=6.1、4.0Hz,1H)、4.39(q,J=3.8 Hz,1H)、4.34(ddd,J=11.8、7.3、4.8Hz,1H)、4.20(ddd,J=11.5、6.8、3.6Hz,1H)、4 .14(q,J=4.0Hz,1H)、3.78(d,J=1.0Hz,6H)、3.51(t,J=3.4Hz,2H)、2.81(ddd,J=14 .1、6.3、4.3Hz,1H)、2.57(dt,J=14.2、6.1Hz,1H)、2.47(ddd,J=13.5、6.3、4.1Hz,1 H)、2.27(dt,J=13.6、6.3Hz,1H)、1.66(d,J=17.7Hz,3H)、0.91(s,9H)、0.12(s,6H) 13 C NMR(151MHz、ァ-d6) δ 163.84, 163.72, 159.72, 159.70, 145.50, 145.33, 145.14, 136.40, 136.30, 134.56, 3.58, 131.01, 130.98, 129.44, 129.41, 129.16, 129.06, 128.78, 127.78, 114.07, 6, 97.10, 87.91, 87.74, 87.68, 86.89, 86.84, 85.91, 85.87, 75.86, 75.82, 72.58, 0, 65.16, 63.36, 55.54, 42.09, 40.78, 40.75, 26.14, 18.48, 12.23, 11.29, -4.54, -4.67 31 P NMR(162MHz。。ツ-d6) δ 33.53 HRMS(ESI-TOF):C60 H 67 NO 13 Calculated value as PSi [M+H] + :1139.4351, Measurement value:1139.4324

[0317] 13. Compound (S p )-55 [ka] (S p )-43 and SI-34 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (10% MeOH in EtOAc) gave 20.3 mg (59%, >20:1 dr) of the title compound (S p )-55 was obtained. Physical state: White amorphous solid R f =0.33(10%MeOH in EtOAc) 1H NMR(600MHz、アセトン-d6) δ 12.08(s,1H)、11.07(s,1H)、8.26(d,J=7.5Hz,1H)、8.20-8.14(m,2H)、7.98(s,1H)、7.70-7.66(m,1H)、7.59(t,J=7.8Hz,2H)、7.52-7.47(m,2H)、7.40-7.32(m,6H)、7.28-7.23(m,1H)、6.92(dd,J=9.0、2.8Hz,4H)、6.36(dd,J=9.0、5.7Hz,1H)、6.27(t,J=6.3Hz,1H)、5.30(tt,J=7.0、3.6Hz,1H)、4.67(d,J=5.2Hz,1H)、4.51-4.43(m,2H)、4.40(dt,J=11.2、6.2Hz,1H)、4.21(t,J=5.0Hz,1H)、3.79(d,J=0.8Hz,6H)、3.53(qd,J=10.7、3.8Hz,2H)、3.00(ddd,J=13.7、9.0、5.2Hz,1H)、2.91-2.84(m,2H)、2.54(dt,J=21.2、6.9Hz,2H)、2.35-2.28(m,2H)、1.64(d,J=17.6Hz,3H)、1.20(d,J=6.8Hz,3H)、1.16(d,J=6.9Hz,3H)、0.94(s,9H)、0.15(d,J=5.4Hz,6H) 13 C NMR(151MHz、アセトン-d6) δ 180.96、159.78、155.87、149.12、148.93、145.52、139.61、136.39、136.29、133.67、131.04、130.99、129.51、129.40、129.09、128.79、127.84、126.50、123.23、114.08、108.73、87.79、87.15、87.10、87.02、86.09、76.39、73.95、66.21、63.49、62.77、55.54、40.81、39.99、36.44、26.17、19.41、19.25、18.57、11.68、10.75、-4.53、-4.60 31 P NMR(162MHz、アセトン-d6) δ 34.67 HRMS(ESI-TOF):C 58 H 69N8O 13 Calculated value as PSi [M+H] + :1145.4569, Measurement value:1145.4559

[0318] 14. Compound (S p )-56 [ka] (S p )-42 and SI-31 were treated on a 0.03 mmol scale according to general procedure I. Purification by pTLC (EtOAc / DCM / acetone = 5:5:3) afforded 13.3 mg (42%, >20:1 dr) of the title compound (S p )-56 was obtained. Physical state: White amorphous solid R f =0.19 (EtOAc / DCM / acetone = 5:5:3) 1H NMR(600MHz、ァ-d6) δ 12.03(s,1H) 10.51(s,1H) 10.17(s,1H) 7.92(s,1H) 7.59(d,J=1.3Hz,1H) 6. 7.44-7.41(m,2H), 7.32-7.29(m,4H), 7.28-7.25(m,2H), 7.23-7.18(m,1H). .86-6.80(m,4H)、6.34(t,J=6.8Hz,1H)、6.25(dd,J=7.9、5.8Hz,1H)、5.50(ddt ,J=8.4、5.6、2.8Hz,1H)、4.58(dt,J=6.3、3.8Hz,1H)、4.33(td,J=4.7、2.7Hz,1 H) 4.24-4.15(m,2H) 3.98(qd,J=3.6;1.1Hz,1H) 3.77(s,6H) 3.38(ddd,J=4 7.0、10.3、4.7Hz,2H)、3.03(ddd,J=13.9、8.0、5.9Hz,1H)、2.89(p,J=6.9Hz,1H). )、2.70(ddd,J=13.9、5.9、2.9Hz,1H)、2.37-2.22(m,2H)、1.92(d,J=1.2Hz,3H) 1.66(d,J=17.7Hz,3H) 1.23(dd,J=6.9;0.8Hz,6H) 0.91(s,9H) 0.12(s,6H) 13 C NMR(151MHz、ァ-d6) δ 179.49, 163.52, 158.30, 158.29, 154.41, 149.87, 147.98, 147.86, 144.50, 136.52, 135.6 3, 135.25, 135.18, 129.66, 129.60, 127.68, 127.25, 126.29, 120.77, 112.54, 109.65, 7, 84.70, 84.66, 84.35, 84.32, 84.16, 82.89, 75.87, 75.83, 71.18, 63.65, 63.60, 4.15, 39.60, 37.65, 35.19, 24.76, 18.02, 17.90, 17.13, 11.46, 11.14, 10.18, -5.91, -6.04 31 P NMR(162MHz。ツ-d6) δ 33.50 HRMS(ESI-TOF):C 52 H66 N7O 13 Calculated value as PSi [M+H] + :1056.4304, Measurement value:1056.4304

[0319] 15. Compound (S p )-57 [ka] (S p )-42 and SI-32 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (1:1:1 EtOAc / DCM / acetone) afforded 25.0 mg (71%, >20:1 dr) of the title compound (S p )-57 was obtained. Physical state: White amorphous solid R f =0.26 (EtOAc / DCM / acetone=1:1:1) 1 H NMR (600 MHz, acetone-d6) δ 11.97(s,1H), 10.73(s,1H), 10.03(s,1H), 8.69(s,1H), 8.51(s,1H), 8.07 (d,J=7.6Hz,2H), 7.90(s,1H), 7.63(t,J=7.4Hz,1H), 7.55-7.49(m,2H), 7 .41-7.36(m,2H), 7.28-7.20(m,6H), 7.19-7.14(m,1H), 6.79(dd,J=9.0, 2 .9Hz,4H), 6.57(t,J=6.4Hz,1H), 6.16(dd,J=7.5, 6.0Hz,1H), 5.35(ddt,J= 8.8, 6.1, 3.2Hz, 1H), 4.95 (q, J=4.9Hz, 1H), 4.28-4.09 (m, 4H), 3.74 (d, J= 1.7Hz,6H), 3.36-3.25(m,2H), 3.12(dt,J=12.8, 6.1Hz,1H), 2.96(ddd,J= 13.6, 7.6, 6.0Hz,1H), 2.84-2.81(m,1H), 2.64-2.53(m,2H), 1.53(d,J=17 .7Hz,3H), 1.19(dd,J=19.1, 6.9Hz,6H), 0.95(s,9H), 0.18(d,J=3.5Hz,6H) 13 C NMR (151 MHz, acetone-d6) δ 180.76, 166.01, 159.59, 155.79, 152.91, 152.68, 151.19, 149.27, 149.09, 145.87, 143.90, 138.06, 136.61, 136.56, 134.78, 133.31, 130.95, 130.91, 129.41, 129.16, 129.00, 128.57, 127.57, 126. 20, 122.22, 113.86, 87.20, 86.55, 86.51, 85.51, 85.47, 84.38, 76.87, 76.83, 72.83, 64.96, 64.92, 64.20, 55.49, 40.42, 38.68, 38.65, 36.53, 26.18, 19.38, 19.23, 18.55, 12.21, 11.26, -4.49, -4.61 31 P NMR (162 MHz, acetone-d6) δ 33.01 HRMS(ESI-TOF):C 59 H 69 N 10 O 12 Calculated value as PSi [M+H] + :1169.4682, Measurement value:1169.4667

[0320] 16. Compound (S p )-58 [ka] (S p )-42 and SI-33 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (5:1 EtOAc / acetone) afforded 17.4 mg (51%, >20:1 dr) of the title compound (S p )-58 was obtained. Physical state: White amorphous solid R f =0.51 (EtOAc / acetone = 5:1) 1H NMR(600MHz、アセトン-d6) δ 12.04(s,1H)、10.68(s,1H)、10.00(s,1H)、8.28(d,J=7.5Hz,1H)、8.10-8.03(m,2H)、7.94(s,1H)、7.66-7.60(m,1H)、7.53(dd,J=8.3、7.3Hz,2H)、7.46-7.38(m,3H)、7.30-7.26(m,4H)、7.26-7.21(m,2H)、7.20-7.15(m,1H)、6.84-6.76(m,4H)、6.30-6.24(m,2H)、5.46(dp、J=8.6、3.0Hz,1H)、4.58(dt,J=6.2、4.3Hz,1H)、4.39-4.34(m,1H)、4.34-4.24(m,2H)、4.14(dt,J=4.7、3.6Hz,1H)、3.75(d,J=0.8Hz,6H)、3.46-3.33(m,2H)、3.08(dt,J=13.7、6.7Hz,1H)、2.73(ddd,J=13.8、6.1、3.2Hz,1H)、2.52(ddd,J=13.5、6.5、4.6Hz,1H)、2.30(dt,J=13.6、6.1Hz,1H)、1.67(d,J=17.6Hz,3H)、1.38(s,1H)、1.22(dd,J=6.9、0.8Hz,6H)、0.91(s,9H)、0.12(d,J=0.9Hz,6H) 13 C NMR(151MHz、アセトン-d6) δ 180.72、163.78、159.61、159.60、155.87、149.33、149.10、145.85、145.44、138.09、136.60、136.53、134.50、133.64、130.98、130.92、129.46、129.09、129.01、128.58、127.60、122.17、113.87、97.20、87.91、87.24、86.77、86.72、85.76、85.73、84.39、77.25、77.22、72.27、65.00、64.96、64.37、55.49、42.23、38.89、38.86、36.54、26.13、19.35、19.31、18.49、12.29、11.35、-4.54、-4.69 31P NMR (162 MHz, acetone-d6) δ 33.52 HRMS(ESI-TOF):C 58 H 69 N8O 13 Calculated value as PSi [M+H] + :1145.4569, Measurement value:1145.4556

[0321] 17. Compound (S p )-59 [ka] (S p )-42 and SI-34 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (EtOAc / acetone / MeOH=5:5:1) afforded 13.4 mg (39%, >20:1 dl) of the title compound (S p )-59 was obtained. Physical state: White amorphous solid R f =0.21 (EtOAc / acetone / MeOH=5:5:1) 1H NMR(600MHz、ァ-d6) δ 12.10(s,2H) 10.82(s,1H) 10.72(s,1H) 8.06(s,1H) 7.92(s,1H) 47-7.42(m,2H), 7.34-7.25(m,6H), 7.24-7.19(m,1H), 6.87-6.81(m,4H). )、6.34(t,J=6.6Hz,1H)、6.22(dd,J=8.3、5.7Hz,1H)、5.37(ddt,J=8.1); 5.3、2.5Hz,1H)、4.75(dt,J=6.1、3.3Hz,1H)、4.37-4.23(m,3H)、4.15(q, J=3.9Hz,1H) 3.78(s,6H) 3.44-3.33(m,2H) 3.05-2.87(m,3H) 2.82(m,1H) 2.68(ddd,J=14.0; 5.7; 2.5Hz,1H) 2.44(ddd,J=13.4; 6.2) 3.7Hz,1H) 1.59(d,J=17.8Hz,3H) 1.25(dd,J=6.8;1.7Hz,6H) 1.17(d ,J=6.8Hz,3H)、1.10(d,J=6.9Hz,3H)、0.95(s,9H)、0.18(d,J=2.9Hz,6H) 13 C NMR(151MHz、ァ-d6) δ 180.92, 180.78, 159.67, 156.16, 155.78, 149.55, 149.46, 149.23, 149.09, 145.84, 138.86, .65, 136.60, 136.50, 131.00, 130.95, 129.01, 128.62, 127.67, 122.57, 122.10, 113.91, 87.31 86.89, 86.85, 85.72, 85.66, 85.63, 84.16, 77.35, 77.31, 73.15, 65.31, 65.27, 64.46, 55.51, .40, 38.94, 38.92, 36.64, 36.43, 26.16, 19.37, 19.26, 19.15, 18.54, 12.08, 11.13, -4.51, -4.65 31 P NMR(162MHz。シ-d6) δ 34.00 HRMS(ESI-TOF):C 56 H 71 N 10O 13 Calculated value as PSi [M+H] + :1151.4787, Measurement value:1151.4779

[0322] 18. Compound (R p )-48 [ka] (R p )-41 and SI-31 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (EtOAc / acetone = 7:1) gave 22.1 mg (69%, >20:1 dr) of the title compound (R p )-48 was obtained. Physical state: White amorphous solid R f =0.31 (EtOAc / DCM / acetone = 5:5:2) 1H NMR(600MHz、アセトン-d6) δ 10.01(s,1H)、9.95(s,1H)、8.57(s,1H)、8.38(s,1H)、8.14(d,J=7.7Hz,2H)、7.69-7.64(m,1H)、7.63(q,J=1.2Hz,1H)、7.58(t,J=7.6Hz,2H)、7.49-7.44(m,2H)、7.36-7.32(m,3H)、7.32- 7.12(m,4H)、6.89-6.80(m,4H)、6.60(dd,J=7.6、6.2Hz,1H)、6.29(dd,J=7.7、6.1Hz,1H)、5.44(ddt,J=8.5、5.8、2.9Hz,1H)、4.60(dt,J=6.2、3.2Hz,1H)、4.41(td,J=4.9、2.6Hz,1H)、4.37-4.28(m,1H)、4.26-4.19(m,1H)、4.09(q,J=4.0Hz,1H)、3.79(s,6H)、3.49-3.44(m,2H)、3.38-3.32(m,1H)、2.82-2.78(m,1H)、2.37-2.30(m,1H)、2.25(ddd,J=13.4、6.2、3.3Hz,1H)、1.81(d,J=1.2Hz,3H)、1.61(d,J=17.6Hz,3H)、0.92(s,9H)、0.14(d,J=2.7Hz,6H) 13 C NMR(151MHz、アセトン-d6) δ 165.94、164.32、159.78、159.76、153.02、152.76、151.41、151.35、146.06、143.61、136.76、136.69、135.21、133.35、132.16、131.12、131.05、129.90、129.78、129.56、129.30、129.18、129.13、128.74、127.74、126.38、126.26、114.05、113.97、111.27、87.39、86.40、86.35、86.14、86.10、85.94、85.61、77.24、77.20、73.24、68.46、65.79、65.75、64.49、55.65、40.81、39.81、38.61、31.32、26.26、24.64、23.78、18.64、14.46、12.63、12.30、11.45、11.35、-4.41、-4.52 31 P NMR (162 MHz, acetone-d6) δ 32.39 HRMS(ESI-TOF):C 55 H 64 N7O 12 Calculated value as PSi [M+H] + :1074.4198, Measurement value:1074.4189

[0323] 19. Compound (R p )-46 [ka] (R p )-40 and SI-33 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (15:1 EtOAc / acetone) afforded 22.0 mg (70%, >20:1 dr) of the title compound (R p )-46 was obtained. Physical state: White amorphous solid R f =0.63 (EtOAc / acetone = 7:1) 1H NMR (600MHz, アセトン-d6) δ 10.08(s,1H), 8.30(d,J=7.5Hz,1H), 8.16-8.12(m,2H), 7.66-7.62(m, 1H), 7.58(d,J=1.3Hz,1H), 7.57-7.52(m,2H), 7.49-7.45(m,2H), 7.41- 7.30(m,7H), 7.26-7.21(m,1H), 6.89(dd,J=8.9,1.7Hz,3H), 6.37(dd,J=8.1,6.1Hz,1H), 6.22(t,J=6.3Hz,1H), 5.35(ddt,J=8.4,5.6,2.7Hz, 1H), 4.57(dt,J=6.2, 4.1Hz,1H), 4.38-4.25(m,3H), 4.18(q,J=4.2Hz, 1H), 3.77(s,6H), 3.43(qd,J=10.6, 3.4Hz,2H), 2.65-2.54(m,2H), 2.48 (ddd,J=13.5,6.3,4.2Hz,1H),2.29(dt,J=13.6,6.3Hz,1H),1.55(d,J=17.7Hz,3H),1.43(d,J=1.2Hz,3H),0.92(s,9H),0.13(d,J=1.5Hz,6H) 13 C NMR (151MHz, Atotron-d6) δ 164.17, 163.70, 159.78, 159.76, 151.28, 145.70, 145.42, 136.44, 136.32, 136.21, 133.57, 131.03, 131.00, 129.42, 129.17, 129.03, 128.77, 127.81, 114.05, 111.29, 8 8.05, 87.71, 86.86, 86.82, 85.39, 85.35, 85.06, 77.07, 77.04, 72.72, 65.43, 65.39, 64.24, 55.55, 42.01, 39.61, 39.58, 26.14, 18.51, 12.18, 12.14, 11.23, -4.51, -4.66 31 P NMR (162MHz, Atom-d6) δ 32.91 HRMS (ESI-TOF):C 54 H 64 N5O 13 PSiとして, calculation value [M+H] +:1050.4086, Measurement value:1050.4081

[0324] 20. Compound (R p )-57 [ka] (R p )-42 and SI-32 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (EtOAc / acetone = 9:2) gave 22.1 mg (63%, >20:1 dr) of the title compound (R p )-57 was obtained. Physical state: White amorphous solid R f =0.29 (EtOAc / acetone = 5:1) 1 H NMR (600 MHz, acetone-d6) δ 11.96 (s, 1H), 10.82 (s, 1H), 10.01 (s, 1H), 8.67 (s, 1H), 8.55 (s, 1H), 8.12 (d, J = 7.6 Hz, 2H), 7.90 (s, 1H), 7.63 (t, J = 7.4 Hz, 1H), 7.53 (t, J = 7.8 Hz, 2H), 7.38-7.33 (m,2H), 7.26-7.19(m,6H), 7.18-7.13(m,1H), 6.81-6.76(m,4H), 6.57(t,J=6.5 Hz,1H), 6.17(t,J=6.3Hz,1H), 5.56(dq,J=9.1, 5.1Hz,1H), 4.95(dt,J=6.2, 4.5H z,1H), 4.38(ddd,J=11.0, 7.0, 3.9Hz,1H), 4.31(dt,J=11.2, 6.2Hz,1H), 4.15(d q,J=16.6, 4.1Hz,2H), 3.74(d,J=0.9Hz,6H), 3.29(d,J=4.3Hz,2H), 3.16(dt,J=1 2.9, 6.2Hz,1H), 2.93-2.89(m,1H), 2.83-2.79(m,1H), 2.59-2.50(m,2H), 1.43( d,J=17.7Hz,3H), 1.14(dd,J=8.5, 6.8Hz,6H), 0.93(s,9H), 0.16(d,J=2.5Hz,6H) 13C NMR (151 MHz, acetone-d6) δ 180.76, 166.10, 159.57, 159.56, 156.00, 152.71, 151.02, 148.97, 145.81, 144.13, 138.61, 136.65, 136.55, 134.69, 133.36, 130.97, 130.92, 130.85, 129.43, 129.21, 128.97, 128.54, 128.52, 127.56, 1 25.99, 122.25, 113.84, 87.06, 86.62, 86.58, 85.48, 84.93, 84.89, 84.33, 75.86, 75.82, 73.04, 65.84, 65.79, 63.63, 55.48, 40.15, 38.51, 36.57, 26.15, 19.39, 19.17, 18.53, 12.07, 11.11, -4.49, -4.64 31 P NMR (162 MHz, acetone-d6) δ 33.13 HRMS(ESI-TOF):C 59 H 69 N 10 O 12 Calculated value as PSi [M+H] + :1169.4682, Measurement value:1169.4670

[0325] 21. Compound (R p )-55 [ka] (R p )-43 and SI-34 were treated according to general procedure I on a 0.03 mmol scale. Purification by pTLC (10% MeOH in EtOAc) gave 16.1 mg (48%, >20:1 dr) of the title compound (R p )-55 was obtained. Physical state: White amorphous solid R f =0.30(10%MeOH in EtOAc) 1H NMR(600MHz、アセトン-d6) δ 12.02(s,1H)、10.86(s,1H)、8.20-8.16(m,2H)、8.14(d,J=7.6Hz,1H)、7.93(s,1H)、7.69-7.63(m,1H)、7.56(t,J=7.8Hz,2H)、7.46-7.41(m,2H)、7.35-7.28(m,5H)、7.22(ddt,J=7.8、6.8、1.3Hz,1H)、6.94-6.87(m,4H)、6.33(dd,J=8.5、6.0Hz,1H)、6.15(t,J=6.3Hz,1H)、5.25-5.18(m,1H)、4.79-4.72(m,2H)、4.36-4.29(m,1H)、4.29-4.18(m,3H)、3.79(d,J=3.0Hz,6H)、3.41(m,2H)、3.00(ddd,J=13.6、8.6、5.4Hz,1H)、2.80-2.74(m,3H)、2.44(dt,J=13.8、6.3Hz,1H)、2.34(ddd,J=13.4、6.1、2.0Hz,1H)、1.62(d,J=17.7Hz,3H)、1.49-1.36(m,1H)、1.18(d,J=6.9Hz,3H)、1.08(d,J=6.8Hz,3H)、0.97-0.86(m,9H)、0.18(d,J=6.5Hz,6H) 13 C NMR(151MHz、アセトン-d6) δ 181.04、168.14、159.91、159.88、156.08、149.03、148.95、145.62、139.61、136.55、136.33、133.74、133.61、132.17、131.17、131.04、129.90、129.78、129.62、129.18、129.14、128.90、127.90、126.26、123.35、114.21、88.08、87.91、87.20、87.15、87.08、86.05、86.01、76.62、74.18、68.46、65.91、65.87、63.73、55.68、40.98、40.07、39.81、36.46、32.76、31.32、27.91、26.32、24.64、23.78、19.60、19.35、18.73、14.46、11.96、11.45、11.01、-4.40、-4.51 31 P NMR (162 MHz, acetone-d6) δ 33.24 HRMS(ESI-TOF):C 58 H 69 N8O 13 Calculated value as PSi [M+H] + :1145.4569, Measurement value:1145.4553

[0326] 22. Compound (R p )-44 [ka] (R p )-40 and SI-31 were treated on a 1.5 mmol scale according to General Procedure I. Purification by silica gel column chromatography (DCM / MeOH=95:5) gave 1.1 g (75%, >20:1 dr) of the title compound (R p )-44 was obtained. Physical state: White amorphous solid R f =0.38 (EtOAc / DCM:acetone=5:5:2) 1H NMR (600MHz, アセトン-d6) δ 10.02(s,1H), 9.96(s,1H), 7.63(q,J=1.2Hz,1H), 7.58(q,J=1.2Hz,1H ), 7.52-7.47(m,2H), 7.40-7.32(m,6H), 7.30-7.24(m,1H), 6.95-6.90( m,4H), 6.39(dd,J=8.3,5.9Hz,1H), 6.30(dd,J=7.6,6.1Hz,1H), 5.33(ddt,J=8.2,5.4,2.6Hz,1H), 4.60(dt,J=6.2,3.2Hz,1H), 4.33-4.27(m,3 H), 4.24(ddd,J=11.4,6.7,4.7Hz,1H),4.11-4.05(m,1H),3.81(s,6H),3.47-3.43(m,2H),2.65-2.53(m,2H),2.33(ddd,J=13.6,7.6,6.1Hz,1 H), 2.25(ddd,J=13.4,6.2,3.3Hz,1H), 1.83(d,J=1.2Hz,3H), 1.55(d,J=17.6Hz,3H), 1.45(d,J=1.2Hz,3H), 0.93(s,9H), 0.15(d,J=1.5Hz,6H) 13 C NMR (151MHz, Atotron-d6) δ 164.32, 164.20, 159.95, 159.93, 151.39, 151.35, 145.80, 136.70, 136.59, 136. 43, 136.25, 131.16, 129.18, 128.92, 127.97, 114.20, 111.44, 111.23, 87.88, 86 .37, 86.33, 85.95, 85.55, 85.51, 85.15, 77.24, 77.20, 73.26, 65.71, 65.67, 64.40, 55.68, 40.83, 39.65, 39.63, 26.26, 18.64, 12.61, 12.24, 11.29, -4.41, -4.52 31 P NMR (162MHz, Atom-d6) δ 32.53 HRMS (ESI-TOF):C 48 H 61 N4O 13 PSiとして, calculation value [M+Na] + :983.3640, measured value: 983.3632

[0327] Example 7 Ψ activation of MPO dimers 1. Obtaining SI-35 for TBS deprotection [ka] Dinucleotide (S p A mixture of SI-44 (290 mg, 0.3 mmol, 1.0 equiv.) and TBAF (1.0 M in THF, 0.75 mL, 0.75 mmol, 2.5 equiv.) in THF (5 mL) was stirred at room temperature for 12 h, after which the mixture was concentrated to dryness under reduced pressure and co-evaporated with toluene (2 × 3 mL). Purification by silica gel column chromatography (DCM / MeOH = 9:1) afforded 223 mg (75%, >20:1 dr) of the title compound SI-35. Physical state: White amorphous solid R f =0.43 (DCM / MeOH=85:15) 1H NMR(600MHz、アセトン-d6) δ 10.05(d,J=15.0Hz,1H)、9.98(d,J=14.4Hz,1H)、7.57(dq,J=13.1、1.2Hz,2H)、7.51-7.46(m,2H)、7.39-7.33(m,4H)、7.33-7.31(m,2H)、7.28-7.23(m,1H)、6.94-6.88(m,4H)、6.36(t,J=7.1Hz,1H)、6.31(dd,J=7.7、6.2Hz,1H)、5.32(dtt,J=10.4、4.2、2.0Hz,1H)、4.59(s,1H)、4.44-4.40(m,1H)、4.27(q,J=3.2Hz,1H)、4.26-4.18(m,1H)、4.08(ddd,J=11.1、7.3、3.3Hz,1H)、4.01(dt,J=6.2、3.3Hz,1H)、3.79(s,6H)、3.46(dd,J=10.6、3.5Hz,1H)、3.42(dd,J=10.6、3.3Hz,1H)、2、2.60-2.55(m,2H)、2.25(ddd,J=13.6、6.2、3.4Hz,1H)、2.19(dddd,J=13.9、7.6、6.2、1.2Hz,1H)、1.82(d,J=1.2Hz,3H)、1.58(d,J=17.7Hz,3H)、1.46(d,J=1.2Hz,3H) 13 C NMR(151MHz、アセトン-d6) δ 164.34、164.26、164.19、159.92、159.90、151.39、151.32、145.87、136.61、136.55、136.49、136.32、131.18、131.15、129.17、128.91、127.95、114.19、111.40、111.22、87.84、86.18、86.14、85.68、85.57、85.53、85.23、76.91、76.87、71.89、71.78、66.26、66.22、64.33、55.68、40.52、40.47、39.67、39.64、12.66、12.25、12.19、11.24 31 P NMR(162MHz、アセトン-d6) δ 33.29 HRMS(ESI-TOF):C 42 H 47 N4O13 P is calculated as [M+Na] + :869.2775, Measurement value:869.2766

[0328] 2. Acquisition of 60 by loading with (+)-CLO-Ψ [ka] SI-35 (170 mg, 0.2 mmol, 1.0 equiv.) and (+)-CLO-Ψ (120 mg, 0.26 mmol, 1.3 equiv.) were dissolved in anhydrous acetonitrile (2 mL, 0.1 M) in a flame-dried round-bottom flask. DBU (39 μL, 0.26 mmol, 1.3 equiv.) was added dropwise to the reaction mixture with stirring. After 30 min, the reaction mixture was quenched by the addition of 1×PBS solution (10 mL) and extracted with DCM (10 mL×3). The combined organic layers were washed with water (10 mL×2) and brine (10 mL), dried over MgSO4, filtered, and concentrated. The remaining crude product was purified by silica gel column chromatography (DCM:MeOH=98:2) to give 168 mg (77%, >20:1 dr) of the title compound 60. Physical state: pale yellow amorphous solid R f =0.48 (DCM:MeOH=95:5) 1H NMR(600MHz、アセトン-d6) δ 7.60(t,J=1.2Hz,1H)、7.55(q,J=1.3Hz,1H)、7.54-7.48(m,2H)、7.41-7.33(m,4H)、7.31-7.22(m,2H)、7.21-7.12(m,1H)、6.96-6.90(m,4H)、6.39(t,J=7.1Hz,1H)、6.29(dd,J=7.9、6.4Hz,1H)、5.34(dddt,J=19.3、11.2、5.6、2.7Hz,2H)、5.02(t,J=1.5Hz,1H)、4.98-4.94(m,1H)、4.54(dt,J=12.8、3.3Hz,1H)、4.36-4.27(m,3H)、4.18(ddd,J=9.7、7.1、2.6Hz,1H)、4.02-3.97(m,1H)、3.81(s,6H)、3.52-3.42(m,2H)、3.29-3.23(m,1H)、2.67(s,1H)、2.60(dd,J=7.2、4.3Hz,2H)、2.51-2.44(m,1H)、2.39-2.34(m,1H)、2.16-2.09(m,1H)、2.03-1.86(m,3H)、1.84(d,J=1.2Hz,3H)、1.82-1.79(m,3H)、1.71(s,3H)、1.63(d,J=17.7Hz,3H)、1.47(d,J=1.2Hz,3H) 13C NMR (151 MHz, acetone-d6) δ 164.16, 159.91, 159.89, 151.31, 151.26, 146.54, 145.83, 138.60, 136.58, 136.56, 136.45, 136.26, 131.16, 131.14, 129.86, 129.18, 129.14, 128.92, 127.96, 126.22, 114.20, 112.24, 111.52, 111.40, 87.87, 87.18, 86.01, 85.57, 85.54, 85.16, 84.33, 84.30, 84.26, 79.38, 79 .33, 77.13, 77.09, 67.11, 65.53, 65.49, 64.41, 64.38, 59.48, 55.70, 54.92, 49.38, 46.32, 44.03, 39.89, 39.72, 39.69, 38.45, 38.41, 34.62, 34.56, 32.77, 28.36, 28.26, 27.36, 27.32, 24.71, 24.52, 24.06, 22.92, 22.15, 21.51, 20.48, 20.23, 13.94, 12.64, 12.28, 12.22, 11.33 31 P NMR (162 MHz, acetone-d6) δ 101.46, 33.22 HRMS(ESI-TOF):C 52 H 62 N4O 14 Calculated value for P2S2Na [M+Na] + :1115.3077, Measurement value:1115.3088

[0329] Example 8 P-Chiral phosphines: Reversed order loading and coupling experiments [ka] Following general procedure C, compound 11b was prepared from 17a in 35% yield and >98:2 er on a 0.05 mmol scale. All physical and spectroscopic data are consistent with 11b synthesized from 8b. Stereochemical confirmation was performed by chiral HPLC analysis (Chiralpak AD-H, isopropanol / hexane = 15:85, 0.8 mL / min, 280 nm) of the reaction product and authentic samples of 11b derived from 8b.

[0330] Example 9 MPO: Reversed order loading and coupling experiments 1. Loading [ka] A flame-dried 50 mL round-bottom flask under argon was charged with the nucleoside (1.0 mmol, 1.0 equiv.), 12a (524.6 mg, 2.0 mmol, 2.0 equiv.), and THF (10 mL). DBU (3.0 mmol, 3.0 equiv.) was added dropwise, and the resulting solution was stirred at room temperature for 12 h. The reaction was then quenched with 20 mL of 1x PBS solution and extracted with DCM (3x30 mL). The organic layers were combined, washed with saturated aqueous NaHCO (10 mL), brine (10 mL), dried over NaSO, filtered, and concentrated. The residue was redissolved in THF (5 mL). EtN (2 equiv.) and MeI (2 equiv.) were added, and the heterogeneous mixture was stirred vigorously for 1 h. The reaction was quenched with 10 mL of water and extracted with EtOAc (3x20 mL). The organic layers were combined, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography to give the desired product SI-36. Physical state: White amorphous solid R f =0.3(EtOH) 1H NMR (600MHz, acetone-d6) δ 10.00(s,1H), 7.54(q,J=1.3Hz,1H), 6.39-6.31(m,1H), 4.66(dt,J=6.4, 3.3Hz,1H), 4.36-4.17(m ,2H), 4.15-4.07(m,1H), 2.40-2.22(m,5H), 1.90-1.79(m,6H), 0.94(s,9H), 0.17(d,J=3.3Hz,6H) 13 C NMR (151 MHz, acetone-d6) δ 164.29, 164.21, 151.37, 136.64, 136.55, 132.16, 129.77, 111.18, 86.37, 86.32, 85.89, 85.72, 73.66, 73.35, 68.45, 65.08, 64.85, 64.81, 40.86, 40.73, 39.80, 31.31, 27.29, 26.26, 24.64, 23.77, 18.83, 18.63, 18.11, 14.46, 12.68, 12.58, 12.29, 12.27, 11.45, -4.43, -4.45, -4.55

[0331] 2. Nucleoside 3'-OH Coupling: [ka] Following general procedure I, 5'-DMTr-dT-3'-OH was coupled with SI-36 to give the desired dT-dT dimer product. 31 P NMR δ 32.57 was reported (R p )dT-dT 31 This corresponds to a P shift (see above). Thus, the stereochemistry at phosphorus of the reaction product is R p It was identified as being

Claims

1. Formula (I): 【Chemistry 1】 [In the formula: R １ is C ６－１０ aryl, where R １ represents one or more of the same or different R ａ may be substituted with a group; R ａ is deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl; R ４ is hydrogen, C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ is alkynyl; R ５ is C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ is alkynyl; R ７ is a substituent that replaces hydrogen, and C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ cycloalkyl, or C ６－１０ aryl; and n is 0, 1, 2, 3, 4, 5, 6, 7, or 8. or a salt thereof.

2. R １ The compound of claim 1 , wherein is phenyl.

3. Formula (II): 【Chemistry 2】 [In the formula: R １ is C ６－１０ Aryl, R ２ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, where R １ and R ２ Each of the groups may be one or more of the same or different R ａ may be substituted with a group; R ａ But deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl. A method for producing a thiophosphinic acid represented by the formula: Organolithium Reagent R ２ Li (where R ２ has the same meaning as above) with a carbanion reagent of formula (Ia): 【Transformation 3】 [In the formula: R １ has the same meaning as above] , a salt thereof, an enantiomer thereof, or a combination thereof to form a thiophosphinic acid of formula (II).

4. The thiophosphinic acid has the formula (IIa): 【Chemistry 4】 4. The method according to claim 3, wherein the compound is a compound represented by the formula:

5. The thiophosphinic acid of formula (II) is reacted with an alkylating reagent selected from methyl iodide, ethyl iodide or propyl iodide to give a compound of formula (III): 【Transformation 5】 [In the formula: R １ is C ６－１０ Aryl, R ２ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, where R １ and R ２ Each of the groups may be one or more of the same or different R ａ may be substituted with a group; R ａ is deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl; and R' is C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ alkynyl] 4. The method of claim 3, further comprising forming a thiophosphinic acid ester of

6. The thiophosphinate ester has the formula (IIIa): 【Transformation 6】 6. The method according to claim 5, wherein the compound is a compound represented by the formula:

7. Thiophosphinic acid esters are 【Transformation 7】 7. The method of claim 6, wherein the compound is selected from the group consisting of:

8. The thiophosphinic ester of formula (III) is reacted with an alkoxide agent, which is an ionic salt consisting of a sodium cation (Na + ) and an alkoxide ( − OR′), to produce a thiophosphinic ester of formula (IV): 【Transformation 8】 [In the formula: R １ is C ６－１０ Aryl, R ２ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, where R １ and R ２ Each of the groups may be one or more of the same or different R ａ may be substituted with a group; R ａ is deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl; and R' is C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ alkynyl] 6. The method of claim 5, further comprising forming an organic phosphinate of the formula:

9. The organic phosphinate is represented by formula (IVa): 【Chemistry 9】 9. The method of claim 8, wherein the compound is a compound represented by the formula:

10. The organic phosphinate is 【Chemistry 10】 10. The method of claim 9, selected from the group consisting of:

11. The organic phosphinate of formula (IV) is reacted with a Grignard reagent R ３ MgBr to form a compound of formula (V): 【Chemistry 11】 [In the formula: R １ is C ６－１０ Aryl, R ２ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, wherein R ３ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, where R １ , R ２ , and R ３ Each of the groups may be one or more of the same or different R ａ may be substituted with a group; R ａ is deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl.

9. The method of claim 8, further comprising forming a phosphine oxide having the formula:

12. The phosphine oxide is represented by formula (Va): 【Chemistry 12】 12. The method of claim 11, wherein the compound is:

13. The phosphine oxide is 【Chemistry 13】 13. The method of claim 12, selected from the group consisting of:

14. 12. The method of claim 11, further comprising reacting the phosphine oxide of formula (V) with a reducing agent to form a phosphine.

15. The phosphine is represented by formula (VI): 【Chemistry 14】 [In the formula: R １ is C ６－１０ Aryl, R ２ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, wherein R ３ is C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl, where R １ , R ２ , and R ３ Each of the groups may be one or more of the same or different R ａ may be substituted with a group; and R ａ is deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl. The method according to claim 14, wherein the compound is

16. The phosphine is represented by formula (VIa): 【Chemistry 15】 16. The method of claim 15, wherein the compound is:

17. Formula (VII) or (VIIa): 【Chemistry 16】 1. A method for preparing an organophosphorus compound loaded with a nucleoside of formula (Ia): 【Chemistry 17】 or an enantiomer thereof, １ wherein: R １ But C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ is alkynyl, R' is C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ is alkynyl, NỮ １ が [Chemistry 18] wherein T is 【Chemistry 19】 and A Ｂｚ but 【Chemistry 20】 and G ｉＢｕ but 【Chemistry 21】 and C Ｂｚ but 【Chemistry 22】 and 【Chemistry 23】 is the bond between each atom.

18. The organophosphorus compound loaded with a nucleoside of formula (VII) or (VIIa) is reacted with a second nucleoside, Nu ２ to react with a compound of formula (VIII) or (VIIIa): 【Chemistry 24】 [In the formula: R １ is C １－６ Alkyl, C ２－６ alkenyl, or C ２－６ is alkynyl; Nu １ and Nu ２ are each independently 【Chemistry 25】 wherein T is 【Chemistry 26】 and A Ｂｚ but 【Chemistry 27】 and G ｉＢｕ but 【Chemistry 28】 and C Ｂｚ but 【Chemistry 29】 and 【Transformation 30】 are the bonds between each atom.

18. The method of claim 17, further comprising forming a dinucleotide represented by

19. a) reacting a dinucleotide of formula (VIII) or (VIIIa) with a compound of formula (Ia) to form a loaded dinucleotide; b) reacting the loaded dinucleotide formed in step (a) with a third nucleoside to form a trinucleotide; c) repeating steps (a) and (b) one or more times to form an oligonucleotide having a desired number of nucleotides.

20. The method of claim 18, further comprising:

20. a) reacting a dinucleotide of formula (VIII) or (VIIIa) with a dinucleotide of formula C: 【Chemistry 31】 or an enantiomer thereof to form a loaded chimeric dinucleotide; and b) reacting the loaded chimeric dinucleotide formed in step (a) with a third nucleoside to form a chimeric trinucleotide bearing phosphonate and phosphorothioate linkages.

20. The method of claim 18, further comprising:

21. Formula (Ia): 【Chemistry 32】 [In the formula: R １ is C ６－１０ aryl, where R １ is one or more of the same or different R ａ may be substituted with a group; R ａ is deuterium, CD ３ , C １－６ Alkyl, C ２－６ Alkenyl, C ２－６ Alkynyl, halogen, CF ３ , C １－６ Alkoxy, C ３－１０ Cycloalkyl, C ６－１０ aryl, or C-linked heteroaryl. or an enantiomer thereof, comprising: Grignard reagent R １ MgBr (where R １ has the same meaning as above) with a carbanion reagent of formula A: 【Transformation 33】 or an enantiomer thereof to form a compound of formula (Ia).

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