Lipid monomers for therapeutic delivery of RNA

Abstract

Provided herein are compounds that contain one or more lipophilic moieties and can be used to prepare one or more lipophilic monomers.Also provided herein are lipophilic monomers that can be conjugated to one or more positions on at least one strand of an oligonucleotide, and oligonucleotides that include at least one of the lipophilic monomers.

Description

[Background technology]

[0001] Efficient delivery of therapeutic RNA beyond the liver is a fundamental obstacle preventing its clinical utility. Conjugate-mediated delivery is emerging as the clinically dominant delivery paradigm for siRNA. Lipids are the major class of conjugates widely used to improve siRNA delivery, as lipid conjugation increases plasma half-life and enhances tissue accumulation and cellular uptake of siRNA. siRNAs are highly hydrophilic and have poor pharmacological properties. Lipid conjugation modulates the hydrophobicity of siRNA, which governs its pharmacokinetic behavior and tissue biodistribution, by driving selective in situ incorporation into the endogenous lipoprotein pathway. While siRNAs have been conjugated to fatty acids, cholesterol, and tocopherol, there remains a need for the development of efficient and robust lipid delivery systems that can balance safety and efficacy for improved therapeutic margins. This application relates to siRNAs conjugated to various lipid classes for delivery of siRNA to tissues other than the liver. Summary of the Invention [Means for solving the problem]

[0002] In one embodiment, provided herein are compounds that contain one or more lipophilic moieties and can be used to prepare one or more lipophilic monomers. In one embodiment, provided herein are lipophilic monomers that can be conjugated, optionally via a linker or carrier, to one or more positions on at least one strand of an oligonucleotide.

[0003] In one embodiment, provided herein is a compound of formula (I):

[0004] [ka] (In the formula, X, Y, D 1 , D 2 , Z1 , Z 2 , Z 3 , R 1 , R 2 , R 3 and n is as defined herein or elsewhere), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0005] In one embodiment, provided herein is a compound of formula (III):

[0006] [ka] (In the formula, X, Y, D 1 , D 2 , Z 2 , R 2 and B are as defined herein or elsewhere), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0007] In one embodiment, provided herein is a compound of formula (IV):

[0008] [ka] wherein G', L', R, and m are as defined herein or elsewhere, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0009] In one embodiment, provided herein is an oligonucleotide comprising at least one lipophilic monomer of the following formula:

[0010] [ka] In the formula, X, Y, Z 1 , Z 2 , Z 3 , R 1 , R 2 , R 3 , B, G', L', R, n, and m are as defined herein or elsewhere.

[0011] Also provided herein are methods of reducing the expression of a target gene in a cell or a subject using the oligonucleotides provided herein. DETAILED DESCRIPTION OF THE INVENTION

[0012] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. All patents, patent applications, published applications and other publications are incorporated herein by reference in their entirety. In the event that there are multiple definitions for terms herein, those in this section prevail unless otherwise stated.

[0013] As used herein and in the appended claims, the indefinite articles "a" and "an" and the definite article "the" include plural as well as singular referents unless the context clearly dictates otherwise.

[0014] As used herein, the terms "comprising" and "including" may be used interchangeably. The terms "comprising" and "including" should be interpreted as specifying the presence of the stated features or components as referred to, but do not exclude the presence or addition of one or more features, components, or groups thereof. Similarly, the terms "comprising" and "including" are intended to include examples encompassed by the term "consisting of." Thus, the term "consisting of" may be used in place of the terms "comprising" and "including" to provide more specific embodiments.

[0015] As used herein, the term "or" should be interpreted as an inclusive "or," meaning any one or any combination. Thus, "A, B, or C" means any of the following: "A, B, C, A and B, A and C, B and C, A, B, and C." Exceptions to this definition will occur only where combinations of elements, features, steps, or acts are inherently mutually exclusive in some way.

[0016] As used herein, the term "and / or" when used in phrases such as "A and / or B" is intended to include both A and B, A or B, A alone, and B alone. Similarly, the term "and / or" when used in phrases such as "A, B, and / or C" is intended to include each of the following embodiments: A, B, and C, A, B, or C, A or C, A or B, B or C, A and C, A and B, B and C, A alone, B alone, and C alone.

[0017] As used herein, unless otherwise specified, the terms "polynucleotide" or "nucleic acid," used interchangeably herein, refer to a polymer of nucleotides of any length, including, for example, DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction. Polynucleotides can include modified nucleotides, such as methylated nucleotides and their analogs. Nucleic acids can be in either single-stranded or double-stranded form. As used herein, unless otherwise specified, "nucleic acid" also includes nucleic acid mimetics (e.g., locked nucleic acids (LNAs), peptide nucleic acids (PNAs), and morpholinos). As used herein, unless otherwise specified, the term "oligonucleotide" refers to a short, generally single-stranded, synthetic polynucleotide, generally, but not necessarily, less than about 200 nucleotides in length. The terms "oligonucleotide" and "polynucleotide" are not mutually exclusive. The above description of polynucleotides is equally fully applicable to oligonucleotides. Unless otherwise specified, the left-hand end of any single-stranded polynucleotide sequence disclosed herein is the 5'-end. The left-hand direction of a double-stranded polynucleotide sequence is referred to as the 5'-direction. The direction of 5'-to-3' addition of nascent RNA transcripts is referred to as the transcription direction. The region of the sequence on the DNA strand that has the same sequence as the RNA transcript at the 5'-to-5' end of the RNA transcript is referred to as the "upstream sequence," and the region of the sequence on the DNA strand that has the same sequence as the RNA transcript at the 3'-to-3' end of the RNA transcript is referred to as the "downstream sequence."

[0018] As used herein, the term "double-stranded RNA agent" or "dsRNA agent" refers to an agent containing an RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecule capable of inhibiting gene expression in a sequence-specific manner. The sense strand and / or antisense strand of a dsRNA agent may include another moiety (e.g., a lipid moiety). For example, a lipid moiety may be incorporated into the sense strand of the dsRNA agent. The sense strand and / or antisense strand of a dsRNA agent may be linked or conjugated directly or indirectly to another moiety (e.g., a lipid moiety). For example, the sense strand of a dsRNA agent may be linked or conjugated directly or indirectly to another moiety (e.g., a lipid moiety). In certain embodiments, a dsRNA agent is a double-stranded RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide comprising a sense strand and an antisense strand that form a double-stranded region. The double-stranded region may be the entire length of the sense strand, the antisense strand, or both. Alternatively, the double-stranded region may be shorter than the entire length of the sense strand, the antisense strand, or both. The double-stranded region can be the result of the antisense strand being completely complementary, partially complementary, or substantially complementary to the sense strand.In certain embodiments, the antisense strand of dsRNA agent is partially complementary to the target RNA transcript.In another specific embodiment, the antisense strand of dsRNA agent is substantially complementary to the target RNA transcript.In another specific embodiment, the antisense strand of dsRNA agent is completely complementary to the target RNA transcript.

[0019] The two strands forming a double-stranded region or duplex structure may be different portions of a larger RNA molecule, or they may be separate RNA molecules. When the two strands are part of a larger RNA molecule and are therefore linked by a continuous chain of nucleotides between the 3' end of one strand and the 5' end of the other strand, forming the double-stranded region or duplex structure, the linked RNA strands are called "hairpin loops." A hairpin loop can contain at least one unpaired nucleotide. In some embodiments, a hairpin loop can contain at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 20, at least 23, or more unpaired nucleotides or nucleotides not directed toward the target site of the dsRNA agent. In some embodiments, a hairpin loop can have 1 to 10 unpaired nucleotides. In some embodiments, a hairpin loop can have 1 to 8 unpaired nucleotides. In some embodiments, a hairpin loop can have 4 to 10 unpaired nucleotides. In some embodiments, a hairpin loop can have 4 to 8 unpaired nucleotides.

[0020] When the two substantially complementary strands of a dsRNA agent are composed of separate RNA molecules, these molecules do not need to be covalently linked, but can be. In certain embodiments where the two strands are covalently linked by means other than a continuous chain of nucleotides between the 3'-end of one strand and the 5'-end of each other strand forming a duplex structure, the connecting structure is called a "linker." The RNA strands of a dsRNA agent may have the same or different number of nucleotides.

[0021] In some embodiments, one or both strands of the dsRNA agent include an overhang. In other embodiments, the dsRNA agent is blunt ended.

[0022] In certain embodiments, the dsRNA agents described herein mediate messenger RNA (mRNA) degradation or mRNA translation inhibition in a sequence-specific manner. In certain embodiments, the dsRNA agents described herein inhibit gene expression via the RNA-induced silencing complex (RISC) pathway. In some embodiments, without being bound by theory, the dsRNA agents described herein function like small interfering RNAs (siRNAs). In certain embodiments, the dsRNA agents described herein are siRNAs.

[0023] As used herein, the term "complementary," when used to describe a first nucleotide sequence (e.g., the sense strand or target sequence of a dsRNA agent) in the context of a second nucleotide sequence (e.g., the antisense strand of a dsRNA agent or a single-stranded antisense oligonucleotide), means that an oligonucleotide or polynucleotide comprising the first nucleotide sequence can hybridize (form base-pair hydrogen bonds under mammalian physiological conditions (or similar in vitro conditions)) to form a duplex or double-helix structure with an oligonucleotide or polynucleotide comprising the second nucleotide sequence under specified conditions. Complementary sequences include Watson-Crick base pairs or non-Watson-Crick base pairs, and also include natural or modified nucleotides or nucleotide mimics, so long as at least the above hybridization requirements are met. Sequence identity or complementarity is independent of modification. For example, fA and mA are complementary to U (or T) and are identical to A for purposes of determining identity or complementarity.

[0024] As used herein, the term "fully complementary" in the context of two nucleotide sequences means that all (100%) of the bases in a contiguous sequence of a first nucleotide sequence hybridize to the same number of bases in a contiguous sequence of a second nucleotide sequence to form a duplex. If two nucleotide sequences are designed to form one or more single-stranded overhangs upon hybridization, such overhangs are not considered mismatches in determining complementarity. For example, a dsRNA agent comprising one oligonucleotide 21 nucleotides in length and another oligonucleotide 23 nucleotides in length, where the 23 nucleotide oligonucleotide comprises a 21 nucleotide sequence that is completely complementary to the 21 nucleotide oligonucleotide, is considered "fully complementary" for the purposes described herein. In certain embodiments, two nucleotide sequences are "fully complementary" when all (100%) of the bases in the first nucleotide sequence hybridize to all (100%) of the bases in the second nucleotide sequence to form a duplex. In one particular embodiment, the two nucleotide sequences hybridize under stringent conditions. In another particular embodiment, the two nucleotide sequences hybridize under very stringent conditions.

[0025] As used herein, the term "partially complementary," in the context of two nucleotide sequences, means that at least 65% but less than 80% of the bases in a contiguous sequence of a first nucleotide sequence hybridize to the same number of bases in a contiguous sequence of a second nucleotide sequence to form a duplex. In one particular embodiment, the two nucleotide sequences hybridize under stringent conditions. In another particular embodiment, the two nucleotide sequences hybridize under very stringent conditions.

[0026] As used herein, the term "substantially complementary" in the context of two nucleotide sequences means that at least 80% but less than 100% of the bases in the contiguous sequence of the first nucleotide sequence hybridize to the same number of bases in the contiguous sequence of the second nucleotide sequence to form a duplex.In some embodiments, two nucleotide sequences are substantially complementary when at least 85%, 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%, but less than 100% of the bases in the contiguous sequence of the first oligonucleotide hybridize to the same number of bases in the contiguous sequence of the second oligonucleotide to form a duplex.In one specific embodiment, the two nucleotide sequences hybridize under stringent conditions.In another specific embodiment, the two nucleotide sequences hybridize under very stringent conditions.

[0027] As used herein, the terms "about" and "approximately," when referring to a numerical value, include the recited numerical value and a variation within ±20%. For example, about 20% includes 16% to 24%, and values ​​therebetween, inclusive. In one embodiment, the terms "about" and "approximately," when referring to a numerical value, include the recited numerical value and a variation within ±15%. In another embodiment, the terms "about" and "approximately," when referring to a numerical value, include the recited numerical value and a variation within ±10%. In another embodiment, the terms "about" and "approximately," when referring to a numerical value, include the recited numerical value and a variation within ±5%.

[0028] As used herein, the term "stringent," when referring to hybridization, means that under "stringent conditions" or "stringent hybridization conditions," a first nucleotide sequence hybridizes to a second nucleotide sequence with minimal hybridization to other sequences. In certain embodiments, an antisense sequence hybridizes to its target sequence under stringent conditions with minimal targeting to other sequences. Stringent conditions are sequence-dependent (e.g., complementary to the length of the sequence) and vary under different environmental parameters (e.g., assay conditions, physiological environment). Examples of stringent hybridization conditions may include 400 mM NaCl, 40 mM PIPES, pH 6.4, 1 mM EDTA, at 50°C or 70°C for 12-16 hours, followed by washing. Those skilled in the art will understand that variations in hybridization stringency are essentially as described.

[0029] As used herein, the term "highly stringent," when referring to hybridization, means that a first nucleotide sequence is only observed to hybridize to a second nucleotide under "highly stringent conditions" or "highly stringent hybridization conditions." In certain embodiments, an antisense sequence is only observed to hybridize to its target sequence under highly stringent conditions. Also, highly stringent conditions may not allow hybridization between partially complementary sequences. Highly stringent conditions are sequence-dependent (e.g., complementary to the length of the sequence) and vary under different environmental parameters (e.g., assay conditions, physiological environment). Highly stringent conditions may include higher temperatures, lower ionic strength, and / or shorter reaction times compared to stringent conditions under the same environment. For example, highly stringent conditions can include hybridization temperatures of about 71° C., about 72° C., about 73° C., about 74° C., about 75° C., about 76° C., about 77° C., about 78° C., about 79° C., about 80° C., or higher. One of skill in the art will understand that variations in hybridization stringency are essentially described.

[0030] As used herein, "target sequence" refers to a contiguous portion of the nucleotide sequence of an RNA molecule formed during transcription of a gene, including mRNAs that are the product of RNA processing of a primary transcript (e.g., mRNAs resulting from alternative splicing). In one embodiment, the contiguous portion of the nucleotide sequence is at least long enough to serve as a substrate for RNAi-directed cleavage at or near that portion of the nucleotide sequence of an mRNA molecule formed during transcription of the gene. In certain embodiments, the target sequence is about 15-30 nucleotides in length. For example, the target sequence may be about 15-30 nucleotides, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20, 15-19, 15-18, 15-17, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 19-30, 19-29, 19-31, 19-32, 19-33, 19-34, 19-35, 19-36, 19-37, 19-38, 19-39, 19-40, 19-41, 19-42, 19-43, 19-44, 19-45, 19-46, 19-47, 19-48, 19-49, 19-50, 19-51, 19-52, 19-53, 19-54, 19-55, 19-56, 19-57, 19-58, 19-59, 19-60, 19-61, 19-62, 19-63, 19-64, 19-65, 19-66, 19-67, 19-68, 19-70, 19-71, 19-72, 19-73, 19-74, 19-75, 1 The target sequence may be 9-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, or 21-22 nucleotides in length. In certain embodiments, the target sequence is 19-25 nucleotides in length. In some embodiments, the target sequence is 21-23 nucleotides in length. Ranges and lengths intermediate to the above-listed ranges and lengths are also contemplated as part of this disclosure.

[0031] As used herein, the phrases "nucleotide sequence corresponding to any one of the antisense strand nucleotide sequences," "nucleotide sequence corresponding to the antisense strand nucleotide sequence," "nucleotide sequence corresponding to any one of the sense strand nucleotide sequences," "nucleotide sequence corresponding to any one of the sense strand nucleotide sequences," or "nucleotide sequence corresponding to any one of the nucleotide sequences" refer to an oligonucleotide comprising a chain of nucleotides containing the listed unmodified nucleotides, or one or more modified nucleotides, or one or more conjugated moieties (e.g., a moiety described herein, such as a lipid, or a modified nucleotide conjugated to a moiety described herein). Those skilled in the art will recognize that the listed unmodified nucleotides can be substituted with other moieties without substantially changing the base pairing properties of the oligonucleotide containing the nucleotide with such a substituted moiety. For example, a nucleotide containing inosine as a base can base pair with a nucleotide containing adenine, cytosine, or uracil. Thus, a nucleotide containing uracil, guanine, or adenine can be replaced, for example, with a nucleotide containing inosine. In another example, adenine and cytosine may be substituted with guanine and uracil, respectively, to form a GU wobble base pair with the target mRNA.

[0032] As used herein, the phrases "non-naturally occurring double-stranded ribonucleic acid (dsRNA) agent," "non-naturally occurring double-stranded ribonucleic acid agent," or "non-naturally occurring dsRNA agent" refer to a dsRNA agent that is not found in nature. A non-naturally occurring dsRNA may contain one or more modified nucleotides.

[0033] As used herein, the term "overhang," in the context of a 5' or 3' nucleotide overhang, refers to at least one unpaired nucleotide protruding from the duplex structure of a dsRNA agent. For example, a nucleotide overhang exists when the 3' end of one strand of a dsRNA agent extends beyond the 5' end of the other strand, or vice versa. In some embodiments, the overhang is present at the 3' end of the sense strand, the antisense strand, or both strands. In one embodiment, the 3'-overhang is present on the antisense strand. In another embodiment, the 3'-overhang is present on the sense strand. In some embodiments, the overhang is present at the 5' end of the sense strand, the antisense strand, or both strands. In one embodiment, the 5'-overhang is present on the antisense strand. In another embodiment, the 5'-overhang is present on the sense strand. The overhang may be due to one strand being longer than the other, or may be the result of two strands of the same length being staggered. In some embodiments, the overhang forms a mismatch with the target sequence. In other embodiments, the overhang is complementary to the targeted gene sequence. Each nucleotide in the overhang region of a dsRNA agent can independently be modified or unmodified (e.g., 2'-fluoro-modified nucleotides, 2'-O-methyl-modified nucleotides, deoxynucleotides, or a combination thereof).

[0034] In some embodiments, the 5'- or 3'-overhang of the sense strand or antisense strand of a dsRNA agent is phosphorylated. In certain embodiments, the 5'- or 3'-overhang of the sense strand and antisense strand of a dsRNA agent is phosphorylated. In some embodiments, the overhang region(s) contain two (or more) nucleotides with a phosphorothioate between the two (or more) nucleotides, and those two (or more) nucleotides can be the same or different.

[0035] In some embodiments, dsRNA agent contains only a single overhang, which can enhance the interference activity of dsRNA agent without affecting its overall stability.For example, the single-stranded overhang can be located at the 3'-end of the sense strand of dsRNA agent, or at the 3'-end of the antisense strand of dsRNA agent.The dsRNA agent can also have a blunt end located at the 5'-end of the antisense strand (or the 3'-end of the sense strand), or vice versa.In some embodiments, the antisense strand of dsRNA agent has a nucleotide overhang at the 3'-end, and the 5'-end is blunt.

[0036] In some embodiments, one strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' end of at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides. In certain embodiments, one strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' end of at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides, but not more than 5 nucleotides. In some embodiments, one strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' end of 1 nucleotide, 2 nucleotides, or 3 nucleotides. In certain embodiments, one strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' end of 1-2 nucleotides, 1-3 nucleotides, 1-4 nucleotides, or 1-5 nucleotides. In some embodiments, one strand of a dsRNA agent includes a 2-3 nucleotide, 2-4 nucleotide, or 2-5 nucleotide overhang at the 5' end, the 3' end, or both the 5' and 3' ends. In certain embodiments, one strand of a dsRNA agent includes a 3-4 nucleotide, or 4-5 nucleotide overhang at the 5' end, the 3' end, or both the 5' and 3' ends. The strand may be the antisense strand or the sense strand. The nucleotide overhang may comprise or consist of a nucleotide analog or a nucleoside analog.

[0037] In some embodiments, each strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' ends of at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides. In certain embodiments, each strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' ends of at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides, but not more than 5 nucleotides. In some embodiments, each strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' ends of 1 nucleotide, 2 nucleotides, or 3 nucleotides. In certain embodiments, each strand of a dsRNA agent includes an overhang at the 5' end, the 3' end, or both the 5' and 3' ends of 1-2 nucleotides, 1-3 nucleotides, 1-4 nucleotides, or 1-5 nucleotides. In some embodiments, each strand of a dsRNA agent includes a 2-3 nucleotide, 2-4 nucleotide, or 2-5 nucleotide overhang at the 5' end, the 3' end, or both the 5' and 3' ends. In certain embodiments, each strand of a dsRNA agent includes a 3-4 nucleotide, or 4-5 nucleotide overhang at the 5' end, the 3' end, or both the 5' and 3' ends. The nucleotide overhangs may comprise or consist of nucleotide analogs or nucleoside analogs.

[0038] As used herein, the term "blunt" or "blunt-ended" in the context of a dsRNA agent means that there are no unpaired nucleotides or nucleotide analogs at a given end of the dsRNA, i.e., there are no nucleotide overhangs. In some embodiments, one end of the dsRNA agent is blunt-ended. In other words, the 5'-end of one strand and the 3'-end of the other strand do not contain unpaired nucleotides or nucleotide analogs. In some embodiments, both ends of the dsRNA agent are blunt-ended. In other words, there are no nucleotide overhangs at either end of the dsRNA agent.

[0039] As used herein, the term "antisense strand" or "guide strand" in the context of a dsRNA agent refers to the strand that includes the region that is complementary to a target sequence.

[0040] As used herein, the term "sense strand" or "passenger strand" in the context of a dsRNA agent refers to the strand of a dsRNA agent that includes a region that is complementary to a region of the antisense strand.

[0041] As used herein, the term "modified" in the context of a nucleobase of a dsRNA agent refers to a nucleobase that is not naturally found in an RNA molecule. Naturally occurring RNA sequences include adenine (A) and guanine (G), as well as the pyrimidine bases cytosine (C) and uracil (U).

[0042] As used herein, the term "modified" in the context of a nucleotide of an RNA sequence, such as a dsRNA agent, refers to a nucleotide that is not naturally found in an RNA molecule.

[0043] As used herein, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency for use in animals, and more particularly for use in humans, or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopoeias.

[0044] In one embodiment, each component is "pharmaceutically acceptable" in the sense of being compatible with the other components of the pharmaceutical composition and suitable for use in contact with the tissues or organs of humans and animals without undue toxicity, irritation, allergic response, immunogenicity, or other problem or complication, commensurate with a reasonable benefit / risk ratio. See, e.g., Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 6th ed., Rowe et al., Eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed., Ash and Ash Eds., Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd ed., Gibson Ed., CRC Press LLC: Boca Raton, FL, 2009. In some embodiments, a pharmaceutically acceptable excipient is nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. In some embodiments, the pharmaceutically acceptable excipient is an aqueous pH buffered solution.

[0045] As used herein, unless otherwise specified, the term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable, relatively non-toxic acids, including inorganic acids and organic acids. In certain embodiments, suitable acids include, but are not limited to, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, citric acid, dihydrogen phosphate, ethenesulfonic acid, fumaric acid, galacturonic acid, gluconic acid, glucuronic acid, glutamic acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, isobutyric acid, isethionic acid, lactic acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, monohydrogencarbonic acid, monohydrogenphosphate, monohydrogensulfuric acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, phthalic acid, propionic acid, suberic acid, succinic acid, sulfuric acid, tartaric acid, toluenesulfonic acid, and the like (see, e.g., S.M. Berge et al., J. Pharm. Sci., 66:1-19 (1977); and Handbook of Pharmaceutical Salts: Properties, Selection and Use, P.H. Stahl and (See C.G. Wermuth, Eds., (2002), Wiley, Weinheim). In certain embodiments, suitable acids are strong acids (e.g., having a pKa of less than about 1), including, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid, pyridinesulfonic acid, or other substituted sulfonic acids. Also included are salts of other relatively non-toxic compounds with acidic properties, including amino acids such as aspartic acid, and other compounds such as aspirin, ibuprofen, and saccharin. Acid addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired acid, either neat or in a suitable solvent. As a solid, the salt can exist in crystalline or amorphous form, or a mixture thereof. The salt can also exist in polymorphic form.

[0046] As used herein, unless otherwise specified, the term "protecting group" refers to a chemical group that blocks a reactive functional group(s) in a compound (such as, but not limited to, carboxy, hydroxy, and amino moieties) from undesired reactions. Protecting groups known to those of skill in the art are provided herein and can be found, for example, in Protective Groups in Organic Synthesis, Greene, TW Buts, PGM, John Wiley & Sons, New York, NY, 5 th Edition, 2014, and can be added or removed using the procedures described herein. Examples of protected hydroxyl groups include silyl ethers, such as those obtained by reacting a hydroxyl group with a reagent, for example, but not limited to, t-butyldiphenylchlorosilane, t-butyldimethylchlorosilane, trimethylchlorosilane, triisopropylchlorosilane, and triethylchlorosilane; substituted methyl and ethyl ethers, for example, but not limited to, methoxymethyl ether, methylthiomethyl ether, benzyloxymethyl ether, t-butoxymethyl ether, 2-methoxyethoxymethyl ether, tetrahydropyranyl ether, 1-ethoxyethyl ether, allyl ether, benzyl ether, and 2-cyanoethyl ether; and esters, for example, but not limited to, benzoylformate, formate, acetate, trichloroacetate, and trifluoroacetate. Examples of protecting groups include, but are not limited to, amidines (e.g., MeN-CH=).

[0047] As used herein, unless otherwise specified, the term "reactive phosphorus group" refers to a chemical group or moiety useful for forming internucleoside linkages, including, for example, phosphodiester and phosphorothioate internucleoside linkages. Such reactive phosphorus groups are known in the art and contain a phosphorus atom in the PIII or PV valence state, including, but not limited to, phosphoramidites, H-phosphonates, phosphate triesters, and phosphorus-containing chiral auxiliaries. In one embodiment, solid-phase synthesis utilizes phosphoramidites (PIII chemistry) as the reactive phosphite. The intermediate phosphite compound is then oxidized to the PV state using known methods to, in some embodiments, generate a phosphodiester or phosphorothioate internucleotide linkage.

[0048] As used herein, and unless otherwise specified, the terms "internucleoside linkage" or "internucleoside linking group" are meant to include all types of internucleoside linking groups known in the art, including, but not limited to, phosphorus-containing internucleoside linking groups such as phosphodiester and phosphorothioate, non-phosphorus-containing internucleoside linking groups such as formacetyl and methylenyllumino, and neutral non-ionic internucleoside linking groups such as 3'-CH2-C(=O)-N(H)-5' or 3'-CH2-N(H)-C(=O)-5'.

[0049] As used herein, unless otherwise specified, the term "alkyl" refers to a saturated, straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms. In one embodiment, an alkyl group is, for example, an alkyl group having 1 to 40 carbon atoms (C1 to C6). 40 alkyl), 1 to 24 carbon atoms (C1 to C 24 alkyl), 4 to 20 carbon atoms (C4 to C 20 alkyl), 6 to 16 carbon atoms (C6 to C 16 alkyl), 6 to 9 carbon atoms (C6 to C9 alkyl), 1 to 15 carbon atoms (C1 to C 15 alkyl), 1 to 12 carbon atoms (C1 to C 12and 1-6 carbon atoms (C1-C6 alkyl), and are attached to the rest of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, pentadecyl, hexadecyl, (1s,3r,5R,7S)-1-heptyl-3-octyladamantane, and the like. Unless otherwise specified, alkyl groups are optionally substituted.

[0050] As used herein, unless otherwise specified, the term "alkenyl" refers to a straight or branched hydrocarbon chain radical, consisting solely of carbon and hydrogen atoms, containing one or more carbon-carbon double bonds. The term "alkenyl" also embraces radicals having "cis" and "trans" configurations, or alternatively, "E" and "Z" configurations, as understood by one of ordinary skill in the art. In one embodiment, an alkenyl group is an alkyl group having, for example, 2 to 40 carbon atoms (C2 to C6). 40 Alkenyl), 2 to 24 carbon atoms (C2 to C 24 Alkenyl, 4 to 20 carbon atoms (C4 to C 20 Alkenyl, 6 to 16 carbon atoms (C6 to C 16 alkenyl), 6 to 9 carbon atoms (C6 to C9 alkenyl), 2 to 15 carbon atoms (C2 to C 15 Alkenyl), 2 to 12 carbon atoms (C2 to C 12 Alkenyl groups have 2 to 8 carbon atoms (C-C alkenyl), 2 to 8 carbon atoms (C-C alkenyl), or 2 to 6 carbon atoms (C-C alkenyl) and are attached to the rest of the molecule by a single bond. Examples of alkenyl groups include, but are not limited to, ethenyl, prop-1-enyl, but-1-enyl, pent-1-enyl, penta-1,4-dienyl, (4Z,7Z,10Z,13Z)-nonadeca-4,7,10,13-tetraene, (4Z,7Z,10Z,13Z)-16,16-dimethylicosa-4,7,10,13-tetraene, and the like. Unless otherwise specified, alkenyl groups are optionally substituted.

[0051] As used herein, unless otherwise specified, the term "alkylene" or "alkylene chain" refers to a linear or branched polyvalent (e.g., divalent or trivalent) hydrocarbon chain that consists solely of carbon and hydrogen, is saturated, and connects the remainder of the molecule to a radical group (or groups). In one embodiment, alkylene refers to, for example, an alkylene having 1 to 24 carbon atoms (C1 to C6). 24 Alkylene, 1 to 15 carbon atoms (C1 to C 15 Alkylene, 1 to 12 carbon atoms (C1 to C 12 Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, n-butylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group(s) can be through one carbon or any two (or more) carbons within the chain. Unless otherwise specified, alkylene chains are optionally substituted.

[0052] As used herein, unless otherwise specified, the term "alkenylene" refers to a straight or branched polyvalent (e.g., divalent or trivalent) hydrocarbon chain, consisting solely of carbon and hydrogen and containing one or more carbon-carbon double bonds, connecting the remainder of the molecule to a radical group (or groups). In one embodiment, an alkenylene is, for example, an alkenylene having 2 to 24 carbon atoms (C2 to C6). 24 Alkenylene, 2 to 15 carbon atoms (C2 to C 15 Alkenylene, 2 to 12 carbon atoms (C2 to C 12Alkenylenes have 2 to 8 carbon atoms (C2-C8 alkenylene), 2 to 6 carbon atoms (C2-C6 alkenylene), or 2 to 4 carbon atoms (C2-C4 alkenylene). Examples of alkenylene include, but are not limited to, ethenylene, propenylene, n-butenylene, and the like. Alkenylene is attached to the rest of the molecule through a single or double bond and to the radical group through a single or double bond. The points of attachment of the alkenylene to the rest of the molecule and to the radical group(s) can be through one carbon or any two (or more) carbons within the chain. Unless otherwise specified, alkenylene is optionally substituted.

[0053] As used herein, unless otherwise specified, the term "cycloalkyl" refers to a saturated non-aromatic monocyclic or polycyclic hydrocarbon radical, consisting solely of carbon and hydrogen atoms. Cycloalkyl groups may include fused, bridged, or spiro ring systems. In one embodiment, cycloalkyl refers to a group having, for example, 3 to 15 ring carbon atoms (C3 to C6). 15 Cycloalkyl), 3 to 10 ring carbon atoms (C3 to C 10 Cycloalkyls have from 1 to 8 ring carbon atoms (C3-C8 cycloalkyl). A cycloalkyl is attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyl radicals include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyl radicals include, but are not limited to, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise specified, when a cycloalkyl radical is fused to an aromatic ring, the resulting fused ring is still considered a cycloalkyl. Unless otherwise specified, cycloalkyl groups are optionally substituted.

[0054] As used herein, unless otherwise specified, the term "cycloalkylene" refers to a polyvalent (e.g., divalent or trivalent) cycloalkyl group. Unless otherwise specified, a cycloalkylene group is optionally substituted.

[0055] As used herein, and unless otherwise specified, the term "heterocyclyl" refers to a non-aromatic radical monocyclic or polycyclic moiety containing one or more (e.g., 1, 1 or 2, 1 to 3, or 1 to 4) heteroatoms independently selected from nitrogen, oxygen, phosphorus, and sulfur. The heterocyclyl may be attached to the main structure at any heteroatom or carbon atom. Heterocyclyl groups can be monocyclic, bicyclic, tricyclic, tetracyclic, or other polycyclic ring systems, where polycyclic ring systems can be fused, bridged, or spiro ring systems. Heterocyclyl polycyclic ring systems can contain one or more heteroatoms in one or more rings. Heterocyclyl groups can be saturated or partially unsaturated. Saturated heterocycloalkyl groups can be referred to as "heterocycloalkyl." A partially unsaturated heterocycloalkyl group can be referred to as a "heterocycloalkenyl" if the heterocyclyl contains at least one double bond, or a "heterocycloalkynyl" if the heterocyclyl contains at least one triple bond. In one embodiment, the heterocyclyl has, for example, 3 to 18 ring atoms (3- to 18-membered heterocyclyl), 4 to 18 ring atoms (4- to 18-membered heterocyclyl), 5 to 18 ring atoms (3- to 18-membered heterocyclyl), 4 to 8 ring atoms (4- to 8-membered heterocyclyl), or 5 to 8 ring atoms (5- to 8-membered heterocyclyl). As used herein, numerical ranges such as "3 to 18" refer to each integer within the given range; for example, "3 to 18 ring atoms" means that the heterocyclyl group can consist of 3 ring atoms, 4 ring atoms, 5 ring atoms, 6 ring atoms, 7 ring atoms, 8 ring atoms, 9 ring atoms, 10 ring atoms, etc., up to 18 ring atoms. Examples of heterocyclyl groups include, but are not limited to, imidazolyl, imidazolidinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyrrolidinyl, furyl, tetrahydrofuryl, thiophenyl, pyridinyl, piperidinyl, quinolyl, and isoquinolyl. Unless otherwise specified, heterocyclyl groups are optionally substituted.

[0056] When groups described herein are said to be "substituted," they may be substituted with any suitable substituent(s). Illustrative examples of substituents include, but are not limited to, those found in the exemplary compounds and embodiments provided herein, as well as halogen atoms such as F, Cl, Br, or I; cyano, oxo (=O); hydroxyl (-OH); alkyl; alkenyl; alkynyl; cycloalkyl; aryl; -(C=O)OR'; -O(C=O)R'; -C(=O)R'; -OR'; -S(O) x R';-S-SR';-C(=O)SR';-SC(=O)R';-NR'R';-NR'C(=O)R';-C(=O)NR'R';-NR'C(=O)NR'R';-OC(=O)NR'R';-NR'C(=O)OR', -NR'S(O) x NR'R';-NR'S(O) x R', and -S(O) x NR'R', where R' at each occurrence is independently H, C1-C 15 alkyl, or cycloalkyl, where x is 0, 1, or 2. In some embodiments, the substituent is C1-C 12 In another embodiment, the substituent is an alkyl group. In another embodiment, the substituent is a cycloalkyl group. In another embodiment, the substituent is a halo group, such as fluoro. In another embodiment, the substituent is a hydroxyl group. In another embodiment, the substituent is an alkoxy group (-OR'). In another embodiment, the substituent is a carboxyl group. In another embodiment, the substituent is an amino group (-NR'R').

[0057] As used herein, unless otherwise specified, the term "optional" or "optionally" (e.g., optionally substituted) means that the subsequently described circumstance event may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted alkyl" means that the alkyl group may or may not be substituted, and that the description includes both substituted alkyl groups and alkyl groups that have no substitution.

[0058] The compounds provided herein contain one or more asymmetric centers and may therefore give rise to enantiomers, diastereomers, and other stereoisomeric forms, which may be defined in terms of absolute stereochemistry as (R)- or (S)-, or as (D)- or (L)-, for amino acids. Unless otherwise specified, the compounds provided herein are meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+)- and (−), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from appropriate optically pure precursors or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, unless otherwise specified, the compounds are intended to include both E and Z geometric isomers, as well as all tautomeric forms.

[0059] As used herein, unless otherwise specified, the term "isomer" refers to different compounds that have the same molecular formula. "Stereoisomers" are isomers that differ only in the arrangement of atoms in space. "Atropisomers" are stereoisomers resulting from hindrance of rotation about a single bond. "Enantiomers" are a pair of stereoisomers that are non-superimposable mirror images of each other. A mixture of a pair of enantiomers in any proportions may be known as a "racemic" mixture. "Diastereoisomers" are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

[0060] "Stereoisomer" can also include E and Z isomers, or mixtures thereof, and cis and trans isomers, or mixtures thereof. In certain embodiments, the compounds described herein are isolated as either the E or Z isomer. In other embodiments, the compounds described herein are a mixture of E and Z isomers.

[0061] It should be noted that if there is a discrepancy between a depicted structure and the name of that structure, the depicted structure takes precedence.

[0062] compound Unless otherwise specified, the descriptions provided herein apply to all formulas provided herein (e.g., formulas (I)-(VIII), including subformulas thereof), to the extent applicable.

[0063] In one embodiment, provided herein is a compound of formula (I):

[0064] [ka] where: X is O or S; Y is O or NR a and D 1 is H, a hydroxyl protecting group, or a reactive phosphorus group; D 2is H, a hydroxyl protecting group, or a reactive phosphorus group; Or, D 1 and D 2 together form a protecting group for both the oxygen and Y to which they are attached, Z 1 , Z 2 , and Z 3 are each independently -O-, -NR a -, * -OC(=O)-, * -NR a C(=O)-, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=S)-, * -NR a C(=S)-, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b -, * -NR a C(=S)NR b -, * -OC(=O)S-, * -NR a C(=O)S-, * -OS(O) x or * -NR a S(O) x - and * points in the direction towards the ring containing X, R 1 , R 2 , and R 3 are each independently H, C1-C6 alkyl optionally substituted with C1-C6 alkoxy, or -(GL) m -R, where R 1 , R 2 , and R 3 At least one of the following is -(GL) m -R, each instance of G is independently C1-C8 alkylene; Each instance of L is independently -O-, -NR a -, -C(=O)-, * -OC(=O)-, * -C(=O)O-, * -NR a C(=O)-, * -C(=O)NR b -, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=S)-, * -C(=S)O-, * -NR a C(=S)-, * -C(=S)NR b -, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b -, * -NR a C(=S)NR b -, * -OC(=O)S-, * -SC(=O)O-, * -NR a C(=O)S-, * -SC(=O)NR b -, * -OS(O) x -, * -S(O) x O-, * -NR a S(O) x -or * -S(O) x NR b - and * points in the direction towards the ring containing X, R a each instance of is independently H or C1-C6 alkyl; R beach instance of is independently H or C1-C6 alkyl; Each instance of R is independently C 12 ~C 32 Alkyl or C 12 ~C 32 alkenyl, or R 1 , R 2 , and R 3 Two or more of the following are -(GL) m -R, one or more R are independently selected from C 12 ~C 32 Alkyl, C 12 ~C 32 alkenyl, or optionally protected mannose; x is 1 or 2, n is 0 or 1, a compound wherein m is 0, 1, or 2; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0065] In one embodiment, X is O. In one embodiment, X is S.

[0066] In one embodiment, Y is O. In one embodiment, Y is NR a In one embodiment, Y is NH.

[0067] In one embodiment, Z 1 , Z 2 , and Z 3 (if present) are each independently -O-, -NR a -, * -OC(=O)-, * -NR a C(=O)-, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -or * -NR a C(=O)NR b In one embodiment, Z 1 , Z 2 , and Z 3(when present) are each independently -O-, -NH-, * -OC(=O)-, * -NHC(=O)-, -OC(=O)O-, * -NHC(=O)O-, * In one embodiment, Z is —OC(═O)NH— or —NHC(═O)NH—. 1 , Z 2 , and Z 3 (if present) are all -O-.

[0068] In one embodiment, n is 0. In one embodiment, the compound is a compound of formula (IA):

[0069] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0070] In one embodiment, the compound is a compound of formula (IA-1):

[0071] [ka] or a pharmaceutically acceptable salt thereof.

[0072] In one embodiment, the compound is a compound of formula (II-A):

[0073] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0074] In one embodiment, the compound is a compound of formula (II-A-1):

[0075] [ka] or a pharmaceutically acceptable salt thereof.

[0076] In one embodiment, n is 1. In one embodiment, the compound is a compound of formula (IB):

[0077] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0078] In one embodiment, the compound is a compound of formula (IB-1):

[0079] [ka] or a pharmaceutically acceptable salt thereof.

[0080] In one embodiment, the compound is a compound of formula (II-B):

[0081] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0082] In one embodiment, the compound is a compound of formula (II-B-1):

[0083] [ka] or a pharmaceutically acceptable salt thereof.

[0084] Unless otherwise specified, when the stereochemistry of a position in a formula provided herein is not specified, the compound of the formula may be any stereoisomer at that position, or a mixture thereof. For example, in one embodiment, the compound of Formula (IB-1) or Formula (II-B-1) is the α-isomer at position 1. In another embodiment, the compound is the β-isomer at position 1, and in yet another embodiment, the compound is a mixture of the α and β isomers.

[0085] In one embodiment, R 1 is H. In one embodiment, R1 is C1-C6 alkyl. In one embodiment, R 1 is methyl. In one embodiment, R 1 is ethyl. In one embodiment, R 1 is n-propyl. In one embodiment, R 1 is isopropyl. In one embodiment, R 1 is n-butyl. In one embodiment, R 1 is n-pentyl. In one embodiment, R 1 is n-hexyl. In one embodiment, alkyl is substituted with C1-C6 alkoxy. In one embodiment, alkoxy is methoxy. In one embodiment, alkoxy is ethoxy. In one embodiment, alkoxy is n-propoxy. In one embodiment, R 1 is 2-methoxyethyl (MOE).

[0086] In one embodiment, R 2 is H. In one embodiment, R 2 is C1-C6 alkyl. In one embodiment, R 2 is methyl. In one embodiment, R 2 is ethyl. In one embodiment, R 2 is n-propyl. In one embodiment, R 2 is isopropyl. In one embodiment, R 2 is n-butyl. In one embodiment, R 2 is n-pentyl. In one embodiment, R 2 is n-hexyl. In one embodiment, alkyl is substituted with C1-C6 alkoxy. In one embodiment, alkoxy is methoxy. In one embodiment, alkoxy is ethoxy. In one embodiment, alkoxy is n-propoxy. In one embodiment, R 2 is 2-methoxyethyl (MOE).

[0087] In one embodiment, R 3 is H. In one embodiment, R 3 is C1-C6 alkyl. In one embodiment, R3 is methyl. In one embodiment, R 3 is ethyl. In one embodiment, R 3 is n-propyl. In one embodiment, R 3 is isopropyl. In one embodiment, R 3 is n-butyl. In one embodiment, R 3 is n-pentyl. In one embodiment, R 3 is n-hexyl. In one embodiment, alkyl is substituted with C1-C6 alkoxy. In one embodiment, alkoxy is methoxy. In one embodiment, alkoxy is ethoxy. In one embodiment, alkoxy is n-propoxy. In one embodiment, R 3 is 2-methoxyethyl (MOE).

[0088] In one embodiment, R 1 is -(GL) m In one embodiment, R 2 is -(GL) m In one embodiment, R 3 is -(GL) m In one embodiment, R 1 and R 2 is -(GL) m In one embodiment, R 1 , R 2 , and R 3 is -(GL) m In one embodiment, R 1 and R 2 is -(GL) m and R 1 The R in the middle is C 12 ~C 32 Alkyl or C 12 ~C 32 alkenyl, and R 2 wherein R is an optionally protected mannose. In one embodiment, R 1 and R 3 is -(GL) m -R and R 1 The R in the middle is C 12 ~C32 Alkyl or C 12 ~C 32 alkenyl, and R 3 In one embodiment, R is an optionally protected mannose. 1 , R 2 , and R 3 is -(GL) m -R and R 1 The R in the middle is C 12 ~C 32 Alkyl or C 12 ~C 32 alkenyl, and R 2 and R 3 wherein R is an optionally protected mannose. In some embodiments, the optionally protected mannose is protected by an acetyl moiety.

[0089] In one embodiment where n is 0, R 1 is -(GL) m -R and R 2 is C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 2 is -(GL) m -R and R 1 is C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 1 and R 2 are each independently -(GL) m -R.

[0090] In one embodiment where n is 1, R 1 is -(GL) m -R and R 2 and R 3 are each independently C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 2 is -(GL) m -R and R 1 and R 3 are each independently C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 3is -(GL) m -R and R 1 and R 2 are each independently C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 1 and R 2 are each independently -(GL) m -R and R 3 is C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 1 and R 3 are each independently -(GL) m -R and R 2 is C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 2 and R 3 are each independently -(GL) m -R and R 1 is C1-C6 alkyl optionally substituted with C1-C6 alkoxy. 1 , R 2 , and R 3 are each independently -(GL) m -R.

[0091] In one embodiment, m is 0. In one embodiment, -(GL) m -R is -R.

[0092] In one embodiment, m is 1. In one embodiment, m is 2.

[0093] In one embodiment, each instance of G is independently C1-C8 alkylene. In one embodiment, each instance of G is independently C1-C4 alkylene. In one embodiment, alkylene is a C1 alkylene. In one embodiment, alkylene is a C2 alkylene. In one embodiment, alkylene is a C3 alkylene. In one embodiment, alkylene is a C4 alkylene. In one embodiment, alkylene is a C5 alkylene. In one embodiment, alkylene is a C6 alkylene. In one embodiment, alkylene is -CH2-. In one embodiment, alkylene is -CH2CH2-. In one embodiment, alkylene is -CH2CH2CH2-.

[0094] In one embodiment, each instance of L is independently -O-, -NH-, * -OC(=O)-, * -C(=O)O-, * -NHC(=O)-, * -C(=O)NH-, -OC(=O)O-, * -NHC(=O)O-, * -OC(=O)NH-, or * In one embodiment, each instance of L is independently: * -NHC(=O)-.

[0095] In one embodiment, -(GL) m -R is -(C2-C6 alkylene)-NH-C(=O)-R. In one embodiment, -(GL) m -R is -(C2-C3 alkylene)-NH-C(=O)-R. In one embodiment, -(GL) m -R is -CH2CH2-NH-C(=O)-R. In one embodiment, -(GL) m -R is -CH2CH2CH2-NH-C(=O)-R.

[0096] In one embodiment, R a is H. In one embodiment, R a is C1-C6 alkyl. In one embodiment, R a is methyl.

[0097] In one embodiment, R b is H. In one embodiment, R b is C1-C6 alkyl. In one embodiment, R b is methyl.

[0098] In one embodiment, provided herein is a compound of formula (III):

[0099] [ka] where: B is a modified or unmodified nucleobase; X is O or S; Y is O or NR a and D 1 is H, a hydroxyl protecting group, or a reactive phosphorus group; D 2 is H, a hydroxyl protecting group, or a reactive phosphorus group; Or, D 1 and D 2 are together protecting groups for both the oxygen and Y to which they are attached, Z 2 -O-, -NR a -, * -OC(=O)-, * -NR a C(=O)-, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=S)-, * -NR a C(=S)-, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b -, * -NR aC(=S)NR b -, * -OC(=O)S-, * -NR a C(=O)S-, * -OS(O) x -or * -NR a S(O) x - and * points in the direction towards the ring containing X, R 2 is -(GL) m -R, Each instance of G is independently C1 to C8 alkylene or C3 to C 10 is cycloalkylene, Each instance of L is independently -O-, -NR a -, -C(=O)-, * -OC(=O)-, * -C(=O)O-, * -NR a C(=O)-, * -C(=O)NR b -, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=S)-, * -C(=S)O-, * -NR a C(=S)-, * -C(=S)NR b -, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b -, * -NR a C(=S)NR b -, * -OC(=O)S-, * -SC(=O)O-, * -NR a C(=O)S-, * -SC(=O)NR b -,* -OS(O) x -, * -S(O) x O-, * -NR a S(O) x -or * -S(O) x NR b - and * points in the direction towards the ring containing X, R a each instance of is independently H or C1-C6 alkyl; R b each instance of is independently H or C1-C6 alkyl; R is C 12 ~C 32 Alkyl or C 12 ~C 32 is alkenyl, x is 1 or 2, m is 0, 1, or 2; However, X is O, Y is O, and Z 2 is -O-, (i) R is -L"-(CH2) 0-3 -R”, where L" is absent, -O-, -NR a -, -C(=O)-, * -OC(=O)-, * -C(=O)O-, * -NR a C(=O)-, * -C(=O)NR b -, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=S)-, * -C(=S)O-, * -NR a C(=S)-, * -C(=S)NR b -, -OC(=S)O-, * -NRa C(=S)O-, * -OC(=S)NR b -, * -NR a C(=S)NR b -, * -OC(=O)S-, * -SC(=O)O-, * -NR a C(=O)S-, * -SC(=O)NR b -, * -OS(O) x -, * -S(O) x O-, * -NR a S(O) x -or * -S(O) x NR b - and * points in the direction towards R, R" is a monocyclic, fused, bridged, or spiro ring moiety, wherein the ring moiety is optionally substituted with one or more C1-C6 alkyl, oxo, or phenyl, wherein the alkyl or phenyl is optionally substituted with one or more halogen, phenyl, or phenoxy, wherein the phenyl or phenoxy is optionally substituted with one or more halogen or C1-C6 alkoxy; (ii) R is C 12 ~C 32 is alkenyl, (iii) m is 1 or 2, and L is -O- or * -C(=O)NR b -or- (iv) m is 1 or 2, and at least one G is C to C 10 a compound which is a cycloalkylene; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0100] In one embodiment, X is O. In one embodiment, X is S.

[0101] In one embodiment, Y is O. In one embodiment, Y is NR aIn one embodiment, Y is NH.

[0102] In one embodiment, the compound is a compound of formula (III-A):

[0103] [ka] or a pharmaceutically acceptable salt thereof.

[0104] In one embodiment, the compound is a compound of formula (III-A-1):

[0105] [ka] or a pharmaceutically acceptable salt thereof.

[0106] In one embodiment, Z 2 is -O-, * -OC(=O)-, * -NR a C(=O)-, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=O)S-, or * -NR a C(═O)S—. In one embodiment, Z 2 is -OC(=O)NH-. In one embodiment, Z 2 is -NHC(=O)-. In one embodiment, Z 2 is -NHC(=O)NH-. In one embodiment, Z 2 is —NHC(═O)S—. In one embodiment, Z 2 is -O-.

[0107] In one embodiment, X is O, Y is O, and Z 2 is -O-, R is C 12 ~C32 In one embodiment, m is 0 and R is C 12 ~C 32 It is alkenyl.

[0108] In one embodiment, X is O, Y is O, and Z 2 is -O-, R is -L"-(CH2) 0-3 In one embodiment, m is 0 and R is substituted with -L"-(CH) 0~3 In one embodiment, m is 0 and R is substituted with fused, bridged, or spirocycloalkyl. In one embodiment, R is substituted with adamantyl. In one embodiment, the adamantyl is 1-adamantyl. In one embodiment, the adamantyl is 2-adamantyl. In one embodiment, R is

[0109] [ka] is replaced by .

[0110] In one embodiment, X is O, Y is O, and Z 2 is -O-, m is 1 or 2, and L is -O- or * -C(=O)NR b In one embodiment, m is 1 and L is * -C(=O)NR b In one embodiment, m is 1 and L is -O-. In one embodiment, X is O, Y is O, and Z is 2 is -O-, then R 2 is -(C2-C6 alkylene)-C(=O)NH-R. In one embodiment, R 2 is -(C2-C3 alkylene)-C(=O)NH-R. In one embodiment, R 2 is —CHCH—C(═O)NH—R. In one embodiment, R 2 is -CH2CH2CH2-C(=O)NH-R. In one embodiment, X is O, Y is O, and Z 2is -O-, then R 2 is -(C2-C6 alkylene)-OR. In one embodiment, R 2 is -(C-C alkylene)-OR. In one embodiment, R 2 is -CHCH-OR. In one embodiment, R 2 is -CH2CH2CH2-OR.

[0111] In one embodiment, X is O, Y is O, and Z 2 is —O—, m is 1 or 2, and at least one G is C 10 In one embodiment, m is 1 and G is C-C 10 It is a cycloalkylene.

[0112] In one embodiment, m is 0. In one embodiment, m is 1. In one embodiment, m is 2.

[0113] In one embodiment, each instance of G is independently C1-C8 alkylene. In one embodiment, each instance of G is independently C1-C4 alkylene. In one embodiment, alkylene is a C1 alkylene. In one embodiment, alkylene is a C2 alkylene. In one embodiment, alkylene is a C3 alkylene. In one embodiment, alkylene is a C4 alkylene. In one embodiment, alkylene is a C5 alkylene. In one embodiment, alkylene is a C6 alkylene. In one embodiment, alkylene is -CH2-. In one embodiment, alkylene is -CH2CH2-. In one embodiment, alkylene is -CH2CH2CH2-.

[0114] In one embodiment, each instance of G is independently C to C 10In one embodiment, the cycloalkylene is cyclopropylene. In one embodiment, the cycloalkylene is cyclobutylene. In one embodiment, the cycloalkylene is cyclopentylene. In one embodiment, the cycloalkylene is cyclohexylene. In one embodiment, the cycloalkylene is cycloheptylene. In one embodiment, the cycloalkylene is cyclooctylene.

[0115] In one embodiment, each instance of L is independently -O-, -NH-, * -OC(=O)-, * -C(=O)O-, * -NHC(=O)-, * -C(=O)NH-, -OC(=O)O-, * -NHC(=O)O-, * -OC(=O)NH-, or * In one embodiment, each instance of L is independently: * -C(=O)NH-.

[0116] In one embodiment, -(GL) m -R is -(C2-C6 alkylene)-C(=O)NH-R. In one embodiment, -(GL) m -R is -(C2-C3 alkylene)-C(=O)NH-R. In one embodiment, -(GL) m -R is -CH2CH2-C(=O)NH-R. In one embodiment, -(GL) m -R is -CH2CH2CH2-C(=O)NH-R.

[0117] In one embodiment, R a is H. In one embodiment, R a is C1-C6 alkyl. In one embodiment, R a is methyl.

[0118] In one embodiment, R b is H. In one embodiment, Rb is C1-C6 alkyl. In one embodiment, R b is methyl.

[0119] In one embodiment, B is adenine (A), guanine (G), thymine (T), cytosine (C), or uracil (U). In one embodiment, B is an unmodified (natural) nucleobase. In one embodiment, B is a modified (natural) nucleobase provided herein or known in the art.

[0120] In one embodiment, D 1 is a hydroxyl protecting group. 1 is 4,4'-dimethoxytrityl chloride (DMTr). In one embodiment, D 1 is 4-monomethoxytrityl (MMTr).

[0121] In one embodiment, D 2 is a reactive phosphorus group. In one embodiment, D 2 teeth,

[0122] [ka] is.

[0123] In one embodiment, D 1 is H and D 2 is H.

[0124] In one embodiment, provided herein is a compound of formula (IV):

[0125] [ka] where: each instance of G' is independently C1-C8 alkylene or C3-C8 cycloalkylene; Each instance of L' is independently * -NR c C(=O)-, *points towards the pyrrolidine-2,5-dione ring, R c Each example is independently H, C1-C 32 Alkyl or C2-C 32 is alkenyl, R is C4~C 32 Alkyl or C4-C 32 is alkenyl, a compound wherein m is 0, 1, or 2; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0126] In one embodiment, m is 0. In one embodiment, m is 1. In one embodiment, m is 2.

[0127] In one embodiment, each instance of G' is independently a C1-C8 alkylene. In one embodiment, each instance of G' is independently a C1-C4 alkylene. In one embodiment, an alkylene is a C1 alkylene. In one embodiment, an alkylene is a C2 alkylene. In one embodiment, an alkylene is a C3 alkylene. In one embodiment, an alkylene is a C4 alkylene. In one embodiment, an alkylene is a C5 alkylene. In one embodiment, an alkylene is a C6 alkylene. In one embodiment, an alkylene is -CH2-. In one embodiment, an alkylene is -CH2CH2-. In one embodiment, an alkylene is -CH2CH2CH2-.

[0128] In one embodiment, each instance of G' is independently a C3-C8 cycloalkylene. In one embodiment, each instance of G' is independently a C4-C6 cycloalkylene. In one embodiment, the cycloalkylene is cyclopropylene. In one embodiment, the cycloalkylene is cyclobutylene. In one embodiment, the cycloalkylene is cyclopentylene. In one embodiment, the cycloalkylene is cyclohexylene. In one embodiment, the cycloalkylene is cycloheptylene. In one embodiment, the cycloalkylene is cyclooctylene.

[0129] In one embodiment, -(G'-L') m -R is

[0130] [ka] In one embodiment, -(G'-L') m -R is

[0131] [ka] is.

[0132] The descriptions of R provided herein apply to all formulas provided herein, to the extent they are applicable.

[0133] In one embodiment, R is C-C 32 In one embodiment, R is C-C alkyl. 32 In one embodiment, R is C-C alkyl. 32 In one embodiment, R is C 10 ~C 32 In one embodiment, R is C 12 ~C 32 In one embodiment, R is C 12 ~C 24 In one embodiment, R is C 14 ~C 18 In one embodiment, R is a C alkyl. In one embodiment, R is a C alkyl. In one embodiment, R is a C alkyl. In one embodiment, R is a C alkyl. 10 In one embodiment, R is C 12 In one embodiment, R is C 14 In one embodiment, R is C 15 In one embodiment, R is C 16 In one embodiment, R is C 17 In one embodiment, R is C 18In one embodiment, the alkyl is a straight chain alkyl. In one embodiment, the alkyl is a branched alkyl.

[0134] In one embodiment, R is C-C 32 In one embodiment, R is C-C alkenyl. 32 In one embodiment, R is C-C alkenyl. 32 In one embodiment, R is C 10 ~C 32 In one embodiment, R is C 12 ~C 32 In one embodiment, R is C 12 ~C 24 In one embodiment, R is C 14 ~C 18 In one embodiment, R is C alkenyl. In one embodiment, R is C alkenyl. In one embodiment, R is C alkenyl. In one embodiment, R is C alkenyl. In one embodiment, R is C 10 In one embodiment, R is C 12 In one embodiment, R is C 14 In one embodiment, R is C 15 In one embodiment, R is C 16 In one embodiment, R is C 17 In one embodiment, R is C 18 In one embodiment, the alkenyl is a straight chain alkenyl. In one embodiment, the alkenyl is a branched alkenyl.

[0135] In one embodiment, R is unsubstituted. In one embodiment, R is -(CH) 13-17 It is CH3.

[0136] In one embodiment, R is -L"-(CH) 0~3 -R”, where L" is absent, -O-, -NR a -, -C(=O)-,* -OC(=O)-, * -C(=O)O-, * -NR a C(=O)-, * -C(=O)NR b -, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b -, * -NR a C(=O)NR b -, * -OC(=S)-, * -C(=S)O-, * -NR a C(=S)-, * -C(=S)NR b -, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b -, * -NR a C(=S)NR b -, * -OC(=O)S-, * -SC(=O)O-, * -NR a C(=O)S-, * -SC(=O)NR b -, * -OS(O) x -, * -S(O) x O-, * -NR a S(O) x -or * -S(O) x NR b - and * points in the direction towards R, R a each instance of is independently H or C1-C6 alkyl; R b each instance of is independently H or C1-C6 alkyl; x is 1 or 2, R'' is a monocyclic, fused, bridged, or spiro ring moiety, wherein the ring moiety is optionally substituted with one or more C1-C6 alkyl, oxo, or phenyl, wherein the alkyl or phenyl is optionally substituted with one or more halogen, phenyl, or phenoxy, wherein the phenyl or phenoxy is optionally substituted with one or more halogen or C1-C6 alkoxy.

[0137] In one embodiment, the ring moiety is C 10 In one embodiment, R" is a cycloalkyl. In one embodiment, R" is a monocyclic cycloalkyl. In one embodiment, R" is a fused cycloalkyl. In one embodiment, R" is a bridged cycloalkyl. In one embodiment, R" is a spirocycloalkyl. In one embodiment, cycloalkyl is a C3-C 12 In one embodiment, the cycloalkyl is cyclohexyl. In one embodiment, the cycloalkyl is bicyclo[2.2.1]heptyl. In one embodiment, the cycloalkyl is adamantyl. In one embodiment, the cycloalkyl is substituted with one or more C1-C6 alkyl. In one embodiment, the cycloalkyl is substituted with one or more methyl or isopropyl.

[0138] In one embodiment, the ring moiety is a 4-10 membered heterocyclyl containing 1-4 heteroatoms independently selected from O, N, and S.

[0139] In one embodiment, R is substituted with -L"-R", where L" is absent, -O-, or -C(=O)O-, and R" is a monocyclic, fused, bridged, or spirocycloalkyl, which is optionally substituted with one or more C1-C6 alkyl. In one embodiment, R is substituted with R". In one embodiment, R is substituted with -OR". In one embodiment, R is substituted with -C(=O)OR". In one embodiment, the substituent is located at the terminal position of R. In one embodiment, R is -(CH2) 14-18 -L”-R”.

[0140] In one embodiment, R″ is

[0141] [ka] In one embodiment, R″ is

[0142] [ka] In one embodiment, R″ is

[0143] [ka] is.

[0144] In one embodiment, R is

[0145] [ka] is replaced by .

[0146] In one embodiment, R is substituted at the terminal position.

[0147] In one embodiment, the compound is a compound of Table 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein B is a modified or unmodified nucleobase.

[0148] [Table 1-1]

[0149] [Table 1-2]

[0150] [Table 1-3]

[0151] [Table 1-4]

[0152] [Table 1-5]

[0153] It should be understood that for compounds in Table 1 having a nucleobase B (e.g., Compound 1), the corresponding compounds in which "B" is replaced by a particular nucleobase provided herein are also specifically provided herein. In one embodiment, specifically provided herein are the corresponding compounds in which "B" is replaced with uracil (U). In one embodiment, specifically provided herein are the corresponding compounds in which "B" is replaced with cytosine (C). In one embodiment, specifically provided herein are the corresponding compounds in which "B" is replaced with adenine (A). In one embodiment, specifically provided herein are the corresponding compounds in which "B" is replaced with guanine (G). In some embodiments, reference to "Table 1" in this application includes these nucleobase-modified compounds.

[0154] Oligonucleotides In one embodiment, provided herein is an oligonucleotide comprising at least one lipophilic monomer of the following formula:

[0155] [ka] In the formula, X, Y, Z 1 , Z 2 , Z 3 , R 1 , R 2 , R 3 , B, G', L', R, n, and m are as defined herein or elsewhere (e.g., as defined in formulas (I), (III), and (IV), respectively, including subformulas thereof).

[0156] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (V):

[0157] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0158] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VA):

[0159] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0160] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VA-1):

[0161] [ka] or a pharmaceutically acceptable salt thereof.

[0162] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-A):

[0163] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0164] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-A-1):

[0165] [ka] or a pharmaceutically acceptable salt thereof.

[0166] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VB):

[0167] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0168] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VB-1):

[0169] [ka] or a pharmaceutically acceptable salt thereof.

[0170] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-B):

[0171] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0172] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-B-1):

[0173] [ka] or a pharmaceutically acceptable salt thereof.

[0174] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VII):

[0175] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0176] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of (VII-A):

[0177] [ka] or a pharmaceutically acceptable salt thereof.

[0178] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of (VII-A-1),

[0179] [ka] or a pharmaceutically acceptable salt thereof.

[0180] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VIII):

[0181] [ka] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0182] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of Table 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, where B is a modified or unmodified nucleobase.

[0183] [Table 2-1]

[0184] [Table 2-2]

[0185] [Table 2-3]

[0186] [Table 2-4]

[0187] For compounds in Table 2 having a nucleobase B (e.g., Compound 1'), it is understood that corresponding compounds in which "B" is replaced by a specific nucleobase provided herein are also specifically provided herein. In one embodiment, specifically provided herein are corresponding compounds in which "B" is replaced with uracil (U). In one embodiment, specifically provided herein are corresponding compounds in which "B" is replaced with cytosine (C). In one embodiment, specifically provided herein are corresponding compounds in which "B" is replaced with adenine (A). In one embodiment, specifically provided herein are corresponding compounds in which "B" is replaced with guanine (G). In some embodiments, references to "Table 2" in this application include these nucleobase-modified compounds.

[0188] In one embodiment, the oligonucleotide is an antisense, antagomir, microRNA, siRNA, pre-microRNA, antimir, ribozyme, RNA activator, U1 adaptor, immune stimulator, or aptamer. In one embodiment, the oligonucleotide is double-stranded RNA (dsRNA). In one embodiment, the oligonucleotide is siRNA.

[0189] In one embodiment, the oligonucleotide is an antisense strand complementary to a target gene; and a sense strand that is complementary to the antisense strand.

[0190] In one embodiment, the lipophilic monomer is present in either the antisense strand or the sense strand. In one embodiment, the lipophilic monomer is present in the sense strand. In one embodiment, the lipophilic monomer is present in the antisense strand.

[0191] In one embodiment, the lipophilic monomer is conjugated to a terminal position (e.g., the 3'-end or the 5'-end) of the oligonucleotide. In one embodiment, the lipophilic monomer is conjugated to the 3'-end of the oligonucleotide. In one embodiment, the lipophilic monomer is conjugated to the 5'-end of the oligonucleotide. In one embodiment, the lipophilic monomer is conjugated to a terminal position (e.g., the 3'-end or the 5'-end) of the sense strand or the antisense strand. In one embodiment, the lipophilic monomer is conjugated to the 3'-end of the sense strand. In one embodiment, the lipophilic monomer is conjugated to the 5'-end of the sense strand. In one embodiment, the lipophilic monomer is conjugated to the 3'-end of the antisense strand. In one embodiment, the lipophilic monomer is conjugated to the 5'-end of the antisense strand.

[0192] In one embodiment, the lipophilic monomer is conjugated directly to the 3' or 5' end of the oligonucleotide. In one embodiment, the lipophilic monomer is conjugated to the 3' or 5' end of the oligonucleotide via a linker group.

[0193] In one embodiment, the lipophilic monomer of formula (V) (or any subformula thereof) is conjugated to a terminal position (e.g., the 3' or 5' end) of the oligonucleotide. In one embodiment, the oligonucleotide comprises the following structure:

[0194] [ka] During the ceremony,

[0195] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation.

[0196] [ka] comprises a linker of 1 to 3 nucleotides at its 3'-end (i.e., O of the lipophilic monomer of formula (V) is directly linked to a nucleotide linker that is linked to the remainder of the oligonucleotide). In one embodiment, the linker is dTdT. In one embodiment, Y is bonded to hydrogen.

[0197] In one embodiment, the oligonucleotide comprises the following structure:

[0198] [ka]

[0199] In one embodiment, the oligonucleotide comprises the following structure:

[0200] [ka]

[0201] In one embodiment, the oligonucleotide comprises the structure:

[0202] [ka] During the ceremony,

[0203] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation.

[0204] [ka] comprises a linker of 1 to 3 nucleotides at the 5'-end (i.e., Y of the lipophilic monomer of formula (V) is directly linked to a nucleotide linker that is linked to the remainder of the oligonucleotide). In one embodiment, the linker is dTdT. In one embodiment, O is bonded to hydrogen.

[0205] In one embodiment, the oligonucleotide comprises the following structure:

[0206] [ka]

[0207] In one embodiment, the oligonucleotide comprises the following structure:

[0208] [ka]

[0209] In one embodiment, the lipophilic monomer of formula (VII) (or any subformula thereof) is conjugated to a terminal position (e.g., the 3' or 5' end) of the oligonucleotide. In one embodiment, the oligonucleotide comprises the following structure:

[0210] [ka] During the ceremony,

[0211] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation.

[0212] In one embodiment, the oligonucleotide comprises the following structure:

[0213] [ka]

[0214] In one embodiment, the oligonucleotide comprises the structure:

[0215] [ka] During the ceremony,

[0216] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation.

[0217] In one embodiment, the oligonucleotide comprises the following structure:

[0218] [ka]

[0219] In one embodiment, the lipophilic monomer of formula (VIII) (or any subformula thereof) is conjugated to a terminal position (e.g., the 3' or 5' end) of the oligonucleotide. In one embodiment, the oligonucleotide comprises the following structure:

[0220] [ka] During the ceremony,

[0221] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation,

[0222] [ka] is the linker.

[0223] In one embodiment, the oligonucleotide comprises the following structure:

[0224] [ka] During the ceremony,

[0225] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation,

[0226] [ka] is the linker.

[0227] In one embodiment, the linker is

[0228] [ka] ("CNH"). In one embodiment, the linker is

[0229] [ka] ("C7NH"). The C6NH or the NH in the C7NH linker is connected to the lipid. In one embodiment, the linker is located at the 5' end of the oligonucleotide. In one embodiment, the linker is located at the 3' end of the oligonucleotide. In one embodiment, the linker is located at an internal position of the oligonucleotide. In one embodiment, the C6NH linker is located at the 5' end of the oligonucleotide. In one embodiment, the C6NH linker is located at the 3' end of the oligonucleotide. In one embodiment, the C6NH linker is located at an internal position of the oligonucleotide. In one embodiment, the C6NH linker is located at the 3' end of the oligonucleotide.

[0230] In one embodiment, the lipophilic monomer is located at an internal position of the oligonucleotide. In one embodiment, the lipophilic monomer is located at an internal position of the sense strand or the antisense strand. In one embodiment, the lipophilic monomer is located at an internal position of the sense strand. In one embodiment, the lipophilic monomer is located at an internal position of the antisense strand.

[0231] In one embodiment, the lipophilic monomer of formula (VII) (or any subformula thereof) is located at an internal position of the oligonucleotide. In one embodiment, the oligonucleotide comprises the structure:

[0232] [ka] wherein, independently,

[0233] [ka] comprises one or more nucleotides of the oligonucleotide in a 5'-3' orientation.

[0234] In one embodiment, the oligonucleotide comprises the following structure:

[0235] [ka]

[0236] In one embodiment, the lipophilic monomer of formula (VIII) (or any subformula thereof) is located at an internal position of the oligonucleotide. In one embodiment, the oligonucleotide comprises the structure:

[0237] [ka] During the ceremony,

[0238] [ka] each independently comprise one or more nucleotides of the oligonucleotide in the 5'-3' orientation.

[0239] In one embodiment, the sense strand and the antisense strand are each 15 to 30 nucleotides in length. In one embodiment, the sense strand and the antisense strand are each 15 to 25 nucleotides in length. In one embodiment, the sense strand and the antisense strand are each 19 to 25 nucleotides in length. In one embodiment, the sense strand and the antisense strand are each 21 to 23 nucleotides in length.

[0240] In one embodiment, the oligonucleotide further comprises a targeting ligand.

[0241] In one embodiment, provided herein is a method of delivering an oligonucleotide to a cell, the method comprising contacting the cell with an oligonucleotide provided herein. In some embodiments, the cell is a brain cell. In some embodiments, the cell is present in a subject, such as a mouse, a non-human primate, or a human. In one embodiment, administration results in an increase in the amount of oligonucleotide delivered to the cell compared to delivery of the same oligonucleotide but without the lipid-conjugate (lipophilic monomer) provided herein. In some embodiments, the increase in the amount of oligonucleotide delivered is determined by an increase in the therapeutic activity of the oligonucleotide.

[0242] In one embodiment, provided herein is a method of reducing expression of a target gene in a cell, the method comprising contacting the cell with an oligonucleotide provided herein. In one embodiment, provided herein is a method of reducing expression of a target gene in a subject, the method comprising administering to the subject an oligonucleotide provided herein.

[0243] In one embodiment, without being limited by any particular theory, oligonucleotides comprising at least one lipophilic monomer provided herein exhibit improved pharmacodynamic profiles over oligonucleotides that do not comprise a lipophilic monomer.

[0244] Chemical modifications to nucleotides In certain embodiments, the dsRNA (e.g., siRNA) agent described herein comprises one or more nucleotide modifications. Nucleotide modifications include, for example, terminal modifications, such as 5'-terminal modifications (e.g., phosphorylation, conjugation, reverse linkage) or 3'-terminal modifications (e.g., conjugation, DNA nucleotide, reverse linkage, etc.); base modifications, such as substitution with a stabilizing base, a destabilizing base, or a base that base-pairs with an expanded repertoire of partners, removal of a base (abasic nucleotide), or conjugated base; sugar modifications (e.g., at the 2' or 4' position) or sugar substitution; or backbone modifications, including modification or substitution of a phosphodiester bond. In some embodiments, the dsRNA agent comprises at least one modification selected from the group consisting of modified internucleoside linkages, modified nucleobases, modified sugars, and any combination thereof. Without limitation, such modifications can be present anywhere in the dsRNA agent (e.g., in the sense strand, the antisense strand, or both strands).

[0245] In some embodiments, a dsRNA agent includes one or more modified sugar modifications, such as one or more substituted sugar moieties. In some embodiments, a dsRNA agent described herein includes one of H, F, or OCH3(OMe) at the 2' position.

[0246] In some embodiments, the dsRNA agents described herein include one or more glycol nucleic acids (GNAs). Typically, GNAs are acyclic nucleic acid analogs whose repeating glycol units are linked by phosphodiester bonds, which differs from the ribose sugar-phosphodiester backbone composition of RNA.

[0247] In certain embodiments, the dsRNA agents described herein include one or more terminal modifications, such as 5'-terminal phosphorylation, conjugation, or inverted linkage. In some embodiments, the terminal modification includes a 5'-phosphate, such as a 5'-terminal phosphate on the antisense strand of the dsRNA agent.

[0248] In some embodiments, the dsRNA agent comprises a sense strand and / or an antisense strand having an inverted abasic nucleotide. In one embodiment, the sense strand contains an inverted abasic nucleotide at the 3' end. In another embodiment, the sense strand contains an inverted abasic nucleotide at the 5' end. In some embodiments, the sense strand contains an inverted abasic nucleotide at both the 5' and 3' ends.

[0249] In some embodiments, the dsRNA agent includes a phosphate or phosphate mimetic at the 5'-end of the antisense strand. In one embodiment, the phosphate mimetic is 5'-vinylphosphonate (VP).

[0250] In some embodiments, the dsRNA agent comprises one or more modified nucleotides. In certain embodiments, the modified nucleotides are selected from the group consisting of 2'O-methyl modified nucleotides, deoxynucleotides, 2'-fluoro modified nucleotides, 2'-O-methyl-uridine, inverted abasic nucleotides, nucleotides containing S-glycol nucleic acids (GNAs), non-locked nucleotides, 5'-vinylphosphonate-2'-O-methyl-uridine, and combinations thereof.

[0251] In some embodiments, the dsRNA agent comprises one or more modified internucleoside linkages (i.e., modified RNA backbones). Modified internucleoside linkages include, for example, phosphorothioates (e.g., phosphoromonothioates). Various salts, mixed salts, and free acid forms are also included. In some embodiments, the dsRNA agent described herein is in free acid form. In other embodiments, the dsRNA agent described herein is in salt form. In one embodiment, the dsRNA agent described herein is in sodium salt form. In certain embodiments, when the dsRNA agent described herein is in salt form, salt cations (e.g., sodium cations) are present in the agent as counterions to substantially all of the electronegative groups (e.g., phosphodiester and / or phosphorothioate groups) present in the agent. In some embodiments, the counterions are condensed counterions. In certain embodiments, the counterions are condensed sodium cations. In some embodiments, the condensed counterions are hydrated. In certain embodiments, the condensed counterions are hydrated sodium cations. A drug in which substantially all of the phosphodiester and / or phosphorothioate bonds have counterions comprises 5, 4, 3, 2, or 1 or less phosphodiester and / or phosphorothioate bonds without counterions.In other words, the electronegative potential of a dsRNA agent is neutralized or substantially neutralized by counterion condensation around the dsRNA.In some embodiments, when the dsRNA agent described herein is in sodium salt form, sodium ions exist around the agent as counterions for substantially all of the phosphodiester and / or phosphorothioate groups present in the agent.

[0252] The phosphate group of internucleoside phosphodiester bond can be modified by replacing one of oxygen atoms with different substituents.One result of this modification can be to improve the resistance of oligonucleotide to nucleotide degradation.Another result of this modification can be to improve the stability of hybridized single-stranded RNA (ssRNA) in dsRNA agent.Examples of modified phosphate group include phosphorothioate (for example, phosphoromonothioate).

[0253] In some embodiments, the dsRNA agent comprises an RNA mimic in which both the sugar and the internucleoside linkage, i.e., the backbone, of the nucleotide unit are replaced with alternative groups. In certain embodiments, the base units are maintained for hybridization with the appropriate target sequence.

[0254] The dsRNA agents described herein can contain one or more asymmetric centers and thus can give rise to enantiomers, diastereomers, and other stereoisomeric forms in terms of absolute stereochemistry, for example, as (R) or (S) for sugar anomers, or as (D) or (L) for amino acids. Included in the dsRNA agents provided herein are all such possible isomers, as well as their racemic and optically pure forms.

[0255] The portion linked to the nucleotide sequence In some embodiments, the dsRNA agents described herein are conjugated to one or more non-nucleotide groups. The non-nucleotide groups can, for example, enhance the targeting, delivery, or conjugation of the dsRNA agent. The non-nucleotide groups can be covalently attached to the 3'-end, 5'-end, and / or internally of either the sense strand and / or antisense strand of the dsRNA agent. The non-nucleotide groups can be covalently attached to the 3'-end, 5'-end, both the 3'-end and 5'-end, internally, both the 3'-end and internally, both the 5'-end and internally, or the 3'-end, 5'-end, and internally of the sense strand and / or antisense strand of the dsRNA agent. In some embodiments, the dsRNA agents described herein contain non-nucleotide groups attached to the 3'-end, 5'-end, both the 3'-end and 5'-end, internally, both the 3'-end and internally, both the 5'-end and internally, or the 3'-end, 5'-end, and internally of the sense strand. In certain embodiments, the dsRNA agents described herein contain a non-nucleotide group linked to the 5'-end of the sense strand. In certain embodiments, the dsRNA agents described herein contain a non-nucleotide group linked to the 3" end of the sense strand. In certain embodiments, the dsRNA agents described herein contain a non-nucleotide group linked within the sense strand. The non-nucleotide group may be linked to the dsRNA agent directly or indirectly via a linker / linking group.

[0256] In some embodiments, a linking group is conjugated to the dsRNA agent. The linking group facilitates covalent attachment of the drug to a targeting ligand or delivery polymer or delivery vehicle. The linking group can be attached to the 3' end, 5' end, and / or internally to the sense strand of the dsRNA agent. In some embodiments, the linking group is attached to the sense strand of the dsRNA agent. In some embodiments, the linking group is conjugated to the 5' end, 3' end, and / or internally to the sense strand of the dsRNA agent. In some embodiments, the linking group is conjugated to the 5' end of the sense strand of the dsRNA agent. In some embodiments, the linking group is conjugated to the 3' end of the sense strand. In some embodiments, the linking group is conjugated internally to the sense strand of the dsRNA agent.

[0257] Typically, a linker or linking group is a connection between two atoms that connects one chemical group (such as a dsRNA agent) or segment of interest to another chemical group (such as a targeting group or a delivery polymer) or segment of interest through one or more covalent bonds.Labile linkages include labile bonds.Linkages can optionally include a spacer that increases the distance between the two bonded atoms.The spacer can further add flexibility and / or length to this linkage.

[0258] The dsRNA agent nucleotide sequences listed in Table 3, whether modified or unmodified, may include 3' or 5' and / or internal targeting ligands and / or linking groups. Any of the dsRNA agent duplexes listed in Tables 1, 2, 3, 4, 5, or 9, whether modified or unmodified, may further include a targeting ligand and / or linking group, which may be attached to the 3' end, the 5' end, and / or internally to either the sense or antisense strand of the dsRNA agent duplex.

[0259] Delivery Vehicle In some embodiments, a delivery vehicle may be used to deliver a dsRNA agent to a cell or tissue.

[0260] composition In one aspect, provided herein is a composition (e.g., a pharmaceutical composition) comprising a dsRNA agent described herein. The composition may further comprise a pharmaceutically acceptable carrier.

[0261] It is understood that the foregoing detailed description and accompanying examples are merely illustrative and should not be construed as limitations on the scope of the subject matter. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications, including but not limited to, with respect to the chemical structures, substituents, derivatives, intermediates, synthesis, formulation, and / or methods of use provided herein, can be made without departing from the spirit and scope thereof. The U.S. patents and publications mentioned herein are incorporated by reference. [Example]

[0262] Further, certain embodiments of the present invention are illustrated by the following non-limiting examples.

[0263] Experimental Procedure for the Synthesis of Oligonucleotide Lipid Conjugates Synthesis was performed by a solid-phase phosphoramidite approach following standard RNA synthesis protocols. All oligonucleotides were prepared on a MerMade 12 synthesizer at a 10 μmole scale using either generic or custom supports. A typical synthesis cycle involves four steps: 1. detritylation, 2. coupling, 3. capping, and 4. oxidation / sulfurization.

[0264] Monomer for RNA phosphoramidites with standard protecting groups, 5'-O-dimethoxytrityl-N 6 -Benzoyl-2'-O-(tert-butyldimethylsilyl)-adenosine 3'-O-(N,N'-diisopropylcyanoethylphosphoramidite), 5'-O-dimethoxytrityl-N 4 -acetyl-2'-O-(tert-butyldimethylsilyl-cytidine-3'-O-(N,N'-diisopropyl-2-cyanoethylphosphoramidite), 5'-O-dimethoxytrityl-N 2-Isobutyryl-2'-O-(tert-butyldimethylsilyl)guanosine 3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite) and 5'-O-dimethoxytrityl-2'-O-(tert-butyldimethylsilyl)uridine 3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite) were used in the synthesis.

[0265] The 2'-modified monomers are 2'-fluorophosphoramidite and 2'-OMe phosphoramidite, 5'-O-dimethoxytrityl-N 4 -acetyl-2'-fluoro-cytidine 3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite), 5'-O-dimethoxytrityl-2'-fluoro-uridine 3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite), 5'-O-dimethoxytrityl-N 6 -Benzoyl-2'-fluoro-adenosine 3'-O-(N,N'-diisopropyl-2-cyanoethylphosphoramidite), 5'-O-dimethoxytrityl-N 2 -Isobutyryl-2'-fluoro-guanosine 3'-O-(N,N'-diisopropyl-2-cyanoethylphosphoramidite), 5'-O-dimethoxytrityl-N 4 -acetyl-2'-OMe-cytidine 3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite), 5'-O-dimethoxytrityl-2'-OMe-uridine-3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite), 5'-O-dimethoxytrityl-N 6 -benzoyl-2'-OMe-adenosine 3'-O-(N,N'-diisopropyl-2-cyanoethyl phosphoramidite), and 5'-O-dimethoxytrityl-N 2 -isobutryl-2'-OMe-guanosine 3'-O-(N,N'-diisopropyl-2-cyanoethylphosphoramidite).

[0266] All phosphoramidites were used at 0.1 M in acetonitrile, except for guanosine, which was used at 0.1 M in 20% DMF / MeCN. Detritylation was performed using 3% dichloroacetic acid in dichloromethane for 3 min. Coupling was performed using a 0.45 M solution of 5-(ethylthio)tetrazole in MeCN, with a coupling time of 12 min. Oxidation of internucleotide phosphites to phosphates was performed using standard 0.02 M iodine in MeCN / pyridine / water (70:20:10). Phosphorothioates were introduced by oxidizing the phosphites to phosphorothioates using a 0.1 M pyridine solution of hydrogenated xanthan. Capping was performed using a THF / acetic anhydride / pyridine (80:10:10) and 1-methylimidazole / acetonitrile solution (20:80 v / v). The complete synthesis cycle took 30 min.

[0267] Lipid Bioconjugation Lipids were incorporated into the sense strand via on-column and / or post-column synthesis. For on-column synthesis, lipids at terminal (3' and 5') and internal positions were synthesized on a solid support. For post-column synthesis, lipids were introduced into the solution phase along with their corresponding N-hydroxysuccinimide esters and the attached amine linker of the nucleotide.

[0268] On-column synthesis For example, on-column introduction of compound 17 onto the 3'-end of the sense strand was achieved using a 3'-amino modifier such as 2-dimethoxytrityloxymethyl-6-fluorenylmethoxycarbonylamino-hexane-1-succinoyl)-long-chain alkylamino-CPGS1-1 (Scheme 1). After deprotection of the Fmoc group with 20% piperidine in DMF, the CPG was thoroughly washed with DMF, MeCN, and diethyl ether and dried. A solution of 80 μmol of compound 17 in 2 mL of 1,4-dioxane was added to the above CPG, followed by 30 μL of DIPEA and shaking for 12 hours. The CPG was washed with DCM, MeCN, and EtO. A solution of 1 mL of CAP A and CAP B was added and shaken for 30 minutes. S1-2 was washed with DCM, MeCN, and EtO, dried, and used for oligo assembly.

[0269] [ka]

[0270] Introduction of compound 17 to the 5'-end of the sense strand was achieved using a 5'-amino modifier such as 6-(4-monomethoxytritylamino)hexyl-(2-cyanoethyl)-(N,N'-diisopropyl)-phosphoramidite. After constructing the sense strand on CPG according to standard procedures, in the penultimate step, the 5'-end of the sense strand was coupled to 6-(4-monomethoxytritylamino)hexyl-(2-cyanoethyl)-(N,N'-diisopropyl)-phosphoramidite. The 4-monomethoxytrityl group was then deblocked using 3% DCA / DCM for 30 min. The CPG was then washed with DCM and ACN and coupled with compound 17 for 12 h (Scheme 2).

[0271] [ka]

[0272] Adamantyl lipids at internal positions were introduced during sense strand assembly using the corresponding phosphoramidite monomers. For example, Scheme 3 shows this using the uridine-based phosphoramidite monomer 25.

[0273] [ka]

[0274] Cleavage, purification, and desalting After completion of the synthesis, the oligonucleotides were cleaved from the support using an ammonia solution at 55°C for 7 hours, with simultaneous deprotection of the nucleobase and phosphate groups. The CPG was filtered and washed with ethanol / acetonitrile / water (3:1:1 v / v). After reducing the volume, the oligonucleotides were purified by reverse-phase or ion-exchange chromatography. The reverse-phase buffer was 0.1 M sodium acetate in 90 / 10% water (Buffer A) and acetonitrile (Buffer B). The ion-exchange buffer was 20 mM sodium phosphate (pH 11) in 90 / 10% water (Buffer A) and 20 mM sodium phosphate (pH 11), 1.8 M sodium bromide in 90 / 10% water (Buffer B). Fractions containing the full-length oligonucleotide were pooled, desalted, and the compounds were analyzed by liquid chromatography-mass spectrometry (LC-MS).

[0275] Post-column synthesis Lipids were bioconjugated at terminal positions (3', 5' ends) and internal positions of the sense strand in solution phase after purification and desalting of the corresponding oligonucleotides.

[0276] For example, incorporation at internal positions was achieved using phosphoramidite monomers derivatized with an amine linker at the 2' position of each nucleotide. Scheme 4 illustrates this using the uridine-based phosphoramidite monomer S4-1. After incorporating S4-1 at the internal position and completing the assembly, cleavage, purification, and desalting processes, the free oligonucleotide bearing a pendant amine was coupled with the corresponding lipid-NHS ester in solution.

[0277] [ka]

[0278] Bioconjugation at terminal / internal positions was carried out using a 0.15 mM solution of deprotected and desalted oligonucleotide in 0.1 M NaHCO3 (pH 8.4) and a 1.5 mM solution of the corresponding lipid-NHS ester in DMF at 60 °C. To a solution of the oligonucleotide (0.25 mL), a DMF solution of the lipid-NHS ester (0.6 mL) was added, and the resulting mixture was heated at 63 °C. Progress was monitored by RP-HPLC (C-8 column; A: 50 mM TEAA, B: MeCN; gradient 5-100% B, 30 °C). The reaction typically reached >90% completion in less than 1 h. The reaction mixture was diluted with water and purified by RP-HPLC (C-8 Xbridge Waters column; A: 50 mM NaOAc, B: MeCN; gradient 5-100% B, 60 °C). The isolated yield of lipid-conjugated oligonucleotide was 60-70%.

[0279] double strand formation For duplex formation, equimolar amounts of sense and antisense strands were mixed and kept at room temperature for 30 minutes. The integrity of the duplex was confirmed by denaturing and non-denaturing HPLC analysis.

[0280] Example 1

[0281] [ka]

[0282] Preparation of 1-2 To a DMF solution (720 mL) of 1-1 (72.0 g, 295.0 mmol) was added TIPDSCl2 (111.0 g, 354.0 mmol) at 0 °C under an inert atmosphere of nitrogen. Imidazole (50.1 g, 737.5 mmol) was then added, and the resulting mixture was stirred at room temperature for 14 h. The mixture was diluted with EtOAc (2.5 L), washed with HO, saturated aqueous sodium bicarbonate, and brine, and dried over Na2SO4. The crude residue was purified by flash column chromatography (SiO2, 0-10% MeOH in DCM) to give 1-2 (115.0 g, 236 mmol, 80% yield) as a white solid. MS: m / z 487 [M+H] + .

[0283] Preparation of 1-3 A mixture of 1-2 (45 g, 92.5 mmol), Na2CO3 (39 g, 370 mmol), and tetrabutylammonium bromide (1.2 g, 3.7 mmol) was dissolved in a biphasic solution of DCM / HO (1.0 / 2.0 L). BzCl (16 mL, 138.7 mmol) was then added to the mixture with vigorous stirring. Vigorous stirring was continued at 30 °C until 2 disappeared on TLC. The mixture was then transferred to a separatory funnel. The organic layer was collected, and the aqueous layer was extracted with DCM. The combined organic extracts were dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel column chromatography (PE / EtOAc 3 / 1). This afforded 1-3 (29.0 g, 53.0% yield) as a white solid. MS: m / z 591 [M+H] + . 1 H NMR (DMSO-d6): δ8.00(d,J=7.4Hz,2H),7.86(d,J=8.0Hz,1H),7.79(t,J=7.4Hz,1H),7.60(t,J=7.8Hz,2H),5.82(d,J=8.0H) z,1H),5.66(d,J=4.4Hz,1H,replaced with D2O),5.57(s,1H),4.21-4.15(m,3H),4.03(m,1H),3.96-3.92(m,1H),1.09-0.99(m,28H).

[0284] Preparation of 1-4 To a solution of 1-3 (10.5 g, 17.8 mmol) and methyl methacrylate (30.5 g, 356 mmol) in t-BuOH (50 mL) was added CsCO (2.9 g, 8.9 mmol). The reaction mixture was stirred at 30 °C for 4 h and then diluted with EtOAc. The mixture was washed twice with H0, dried over NaSO, and concentrated. The evaporated residue was purified by column chromatography (SiO, PE / EtOAc 6 / 1) to give 1-4 (8.7 g, 72% yield) as a white solid. MS: m / z 677 [M+H] + .

[0285] Preparation of 1-5 To a solution of 1-4 (8.7 g, 12.8 mmol) in MeOH (87 mL) was added K2CO3 (3.5 g, 25.6 mmol). The reaction mixture was stirred at 30 °C for 2 h. It was then diluted with EtOAc / water (1 / 1), and the organic layer was washed with water, dried over Na2SO4, and concentrated. The residue was purified by flash preparative HPLC to give (C 18 Column, mobile phase CHCN and HO (0.5% NHHCO), gradient 50% to 100% MeCN), afforded 1-5 (6.1 g, 82%) as a white solid. MS: m / z 573 [M+H] + .

[0286] Preparation of 1-6 To a solution of 1-5 (6.1 g, 10.6 mmol) in THF (60 mL) was added TBAF (12.7 mL, 12.7 mmol). The reaction mixture was stirred at room temperature for 1 h, then concentrated, and the residue was purified by column chromatography (SiO, 0-5% MeOH in EtOAc). This afforded 1-6 (3.2 g, 90% yield) as a white solid. MS: m / z 331 [M+H] + ; 1H NMR (DMSO-d): δ 11.32 (s, 1H, exchanged with DO), 7.91 (d, J = 8.0 Hz, 1H), 5.81 (d, J = 5.0 Hz, 1H), 5.65 (d, J = 8.0 Hz, 1H), 5.13 (s, 1H, exchanged with DO), 5.03 (d, J = 5.0 Hz, 1H, exchanged with DO), 4.10 (m, 1H), 3.92 (m, 1H), 3.84-3.70 (m, 3H), 3.65-3.53 (m, 3H), 2.56 (m, 2H).

[0287] Preparation of 1-7 To a solution of 1-6 (3.2 g, 9.6 mmol) in THF / HO (30 / 15 mL) was added KOH (1.6 g, 28.8 mmol). The reaction mixture was stirred at 20 °C for 1 h. The solution was then heated to 100 °C for 1 h. + Neutralization to pH 7 with exchange resin, filtration, and concentration gave 1-7 as a white solid. MS: m / z 317 [M+H] + .

[0288] Preparation of 1-8 To a solution of 1-7 (3.0 g, 9.6 mmol) in DMF (30 mL) were added EDCI (1.8 g, 9.6 mmol), HOBT (1.3 g, 9.6 mmol), hexadecan-1-amine (2.3 g, 9.6 mmol), and DIPEA (2.5 g, 19.2 mmol). The reaction mixture was stirred at 20 °C for 24 h and diluted with EtOAc. The mixture was washed with HO and 5% citric acid, dried over NaSO, and concentrated. The residue was purified by precipitation with EtOAc to give 1-8 (4.7 g, 78% yield over two steps) as a white solid. MS: m / z 540 [M+H] + . 1 H NMR (DMSO-d): δ 11.32 (s, 1H, exchanged with DO), 7.91 (d, J = 8.0 Hz, 1H), 7.88 (t, J = 5.2 Hz, 1H, exchanged with DO), 5.81 (d, J = 5.3 Hz, 1H), 5.65-5.62 (dd, J = 8.0, 1.8 Hz, 1H), 5.16 (s, 2H, exchanged with DO), 4.17 (m, 1H), 3.94-3.54 (m, 6H), 3.00 (m, 2H), 2.34 (m, 2H), 1.37-1.17 (m, 28H), 0.87-0.82 (m, 3H).

[0289] Preparation of 1-9 To a solution of 1-8 (3.2 g, 5.9 mmol) in pyridine (30 mL) was added DMTrCl (2.4 g, 7.1 mmol) under N2 atmosphere. The reaction mixture was stirred at room temperature for 3 h. The reaction mixture was then diluted with EtOAc, washed with HO, and dried over Na2SO4. The evaporated residue was purified by silica gel column chromatography (PE / EtOAc 2 / 1 to 1 / 2) to give 1-9 (3.3 g, 62% yield) as a white solid. MS: m / z 840 [M−H] - ; 1 H-NMR (DMSO-d): δ 11.38 (s, 1H, exchanged with DO), 7.87 (t, J = 5.2 Hz, 1H, exchanged with DO), 7.70 (d, J = 8.0 Hz, 1H), 7.39-7.21 (m, 9H), 6.91-6.89 (m, 4H), 5.77 (d, J = 3.4 Hz, 1H), 5.29 (d, J = 8.0 Hz, 1H) ),5.26(d,J=6.3Hz,1H,replaced with D2O),4.24(m,1H),4.03-3.92(m,2H),3.79(m,2H),3.74(s,6 H),3.30-3.20(m,2H),3.01(m,2H),2.37(m,2H),1.38-1.17(m,28H),0.87-0.81(m,3H).

[0290] Preparation of 1 To a solution of 1-9 (2.9 g, 3.4 mmol) in DCM (30 mL) were added DCI (418 mg, 2.9 mmol) and CN(CH2)2OP[N(iPr)2]2 (1.2 g, 4.1 mmol) under a N2 atmosphere. The reaction mixture was stirred at 30 °C for 2 h. The solution was then diluted with DCM and washed with H2O. The combined organic layers were washed with brine, dried over Na2SO4, and concentrated. The residue was purified by flash preparative HPLC (C 18 Purification by column chromatography (mobile phase H2O containing 0.5% NH4HCO3, CH3CN; gradient 0-100% MeCN) gave compound 1 (2.6 g, 71%) as a white solid. LCMS: m / z 1040 [MH] - ; 1H-NMR(DMSO-d6): δ11.38(s,1H),7.79-7.72(m,2H),7.40-7.21(m,9H),6.91-6.86 (m,4H),5.80(m,1H),5.30-5.23(dd,J=19.6,8.0Hz,1H),4.44-4.32(m,1H),4.13- 4.05(m,2H),3.86-3.46(m,12H),3.36-3.25(m,2H),2.98(m,2H),2.78(m,1H),2.6 1(m,1H),2.33(m,2H),1.35-1.17(m,28H),1.15-0.95(m,12H),0.86-0.83(m,3H); 31 P-NMR (DMSO-d6): δ149.22,148.69.

[0291] Example 2.

[0292] [ka]

[0293] Preparation of 2-1 To a solution of 1-2 (9.7 g, 19.9 mmol) in anhydrous DCM (100 mL) was added CDI (3.5 g, 19.9 mmol), and the reaction mixture was stirred at room temperature for 1 hour. The mixture was used in the next step without purification. ESI-LCMS: m / z 581 [M+H] + .

[0294] Preparation of 2-2 To a solution of 2-1 (11.5 g, 19.9 mmol) in anhydrous DCM (100 mL) was added hexadecan-1-amine (4.8 g, 19.9 mmol) in anhydrous DCM (100 mL) dropwise. The reaction mixture was stirred at 30 °C for 16 h, then quenched with HO and diluted with DCM. The organic phase was washed with HO and 5% citric acid and dried over NaSO. The evaporated residue was purified by silica gel column chromatography (0-20% acetone in 1 / 1 EtOAc / DCM) to give 2-2 (11.1 g, 70% yield for two steps) as a white solid. MS: m / z 754 [M+H] + ; 1H NMR (DMSO-d6): δ11.41(s,1H,exchanged with D2O),7.69(d,J=8.0Hz,1H),7.33(t,J=5.5Hz,1H,exchanged with D2O),5.66(d,J=1.6Hz,1H),5.59(d,J=8.0Hz,1H),5.33-5. 31(m,1H),4.51-4.47(m,1H),4.09-4.05(m,1H),3.95-3.82(m,2H),3.05 -2.85(m,2H),1.37-1.22(m,28H),1.05-0.96(m,24H),0.87-0.82(m,3H).

[0295] Preparation of 2-3 To a solution of 2-2 (4.7 g, 6.2 mmol) in THF (50 mL) was added 1 M TBAF in THF (7.4 mL, 7.4 mmol). The reaction mixture was stirred at room temperature for 1 h. EtOAc was added, and the mixture was washed with HO and dried over NaSO. The residue was purified by precipitation from EtOAc to give 2-3 (3.1 g, 92% yield) as a white solid. MS: m / z 512 [M+H] + ; 1 H NMR (DMSO-d): δ 11.33 (s, 1H, exchanged with DO), 7.90 (d, J = 8.0 Hz, 1H), 7.27 (t, J = 5.5 Hz, 1H, exchanged with DO), 5.97 (d, J = 6.0 Hz, 1H), 5.66 (d, J = 8.0 Hz, 1H), 5.46 (d, J = 4.9 Hz, 1H, exchanged with DO), 5.19 (m, 1H, exchanged with DO), 5.01 (t, J = 5.7 Hz, 1H), 4.19 (m, 1H), 3.88 (m, 1H), 3.62-3.57 (m, 2H), 2.91 (m, 2H), 1.37-1.17 (m, 28H), 0.87-0.82 (m, 3H).

[0296] Preparation of 2-4 To a solution of 2-3 (3.1 g, 6.0 mmol) in pyridine (30 mL) was added DMTrCl (2.2 g, 6.6 mmol) under a N atmosphere. The reaction mixture was stirred at room temperature for 3 h, then diluted with EtOAc, washed with HO, and dried over NaSO. The residue was purified by silica gel column chromatography (PE / EtOAc 2 / 1 to 1 / 2) to give 2-4 (3.1 g, 60% yield) as a yellow solid. MS: m / z 812M-H - ; 1 H-NMR (DMSO-d6): δ11.39 (s, 1H, exchanged with D2O), 7.71 (d, J=8.0Hz, 1H), 7.39-7.21 (m ,10H),6.91-6.89(m,4H),5.91(d,J=4.8Hz,1H),5.52(d,J=5.6Hz,1H,replaced with D2O) ,5.37(d,J=8.0Hz,1H),5.14(t,J=5.1Hz,1H),4.35(m,1H),3.96(m,1H),3.74( s,6H),3.32-3.19(m,2H),2.95(m,2H),1.38-1.15(m,28H),0.86-0.83(m,3H).

[0297] Preparation of 2 To a solution of 2-4 (1.9 g, 2.3 mmol) in DCM (20 mL) was added DCI (234 mg, 1.9 mmol) and CN(CH)OP[N(iPr)] (843 mg, 2.8 mmol) under a N atmosphere. The reaction mixture was stirred at 30 °C for 2 h. The solution was then diluted with DCM and washed with HO. The organic layer was washed with brine, dried over NaSO, and evaporated. The residue was purified by silica gel column chromatography (PE / EtOAc 2 / 1-1 / 2) to give compound 2 (2.1 g, 86% yield) as a white solid. MS: m / z 1012 [M−H] - ; 1H-NMR (DMSO-d6): δ11.25(s,1H),7.67(d,J=8.0Hz,1H),7.43-7.21(m,10H),6.90-6.87(m,4H),5.97(m,1H),5.42-5.32(m,2H),4.60(m,1H) ,4.20-4.14(m,2H),3.81-3.34(m,12H),2.98(m,2H),2.73(m,1H),2. 58(m,1H),1.43-1.19(m,28H),1.15-1.00(m,12H),0.86-0.83(m,3H); 31 P-NMR (DMSO-d6): δ149.64,149.60.

[0298] Example 3

[0299] [ka]

[0300] Preparation of 3-3 A suspension of 3-1 (31.0 g, 137.1 mmol), DMTrCl (51.0 g, 150.8 mmol), and DMAP (0.2 g, catalytic amount) in a mixture of pyridine (110 mL) and DMF (78 mL) was stirred at room temperature for 15 h and then concentrated. The residue was partitioned between dichloromethane and water. The organic layer was washed with aqueous sodium bicarbonate and brine and dried over MgSO. The evaporated residue was co-evaporated with toluene to give compound 3-2 as a viscous oil. CClCN (140.0 g) and TEA (3 mL) were added, and the solution was heated at 90 °C for 16 h. The black reaction mixture was concentrated, and the residual oil was purified by silica gel column chromatography (DCM:MeOH 100 / 1 to 20 / 1) to give compound 3-3 as a foam (89.1 g, 97% yield). MS: m / z 670 [M−H] - .

[0301] Preparation of 3-4 To a solution of 3-3 (89.1 g, 132.8 mmol) in ethanol (360 mL) was added 6N NaOH solution (180 mL), and the reaction was refluxed at 70 °C for 16 h, then cooled and concentrated. The residue was partitioned between dichloromethane and saturated ammonium chloride. The aqueous phase was washed with dichloromethane, dried over combined magnesium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (DCM containing 1% TEA / MeOH 30 / 1 to 10 / 1) to give compound 3-4 (42.0 g, 77.1 mmol, 56.2% yield over three steps) as a white foam. MS: 544 m / z [M−H] - . 1 H NMR(DMSO-d6)δ7.62(dd,J=8.3,4.8Hz,1H),7.43-7.19(m,9H),6.97-6.83(m,4H),5.79-5.61(m,2H),5. 39(dd,J=8.0,2.7Hz,1H),3.98(d,J=7.6Hz,2H),3.74(s,6H),3.42-3.11(m,4H),2.60(q,J=7.2Hz,2H).

[0302] Preparation of 3-5 To a solution of 3-4 (4.2 g, 7.7 mmol) in dry DMF (42 mL) was added palmitic acid (2.0 g, 7.7 mmol), EDCI (1.5 g, 7.7 mmol), HOBT (1.1 g, 7.7 mmol), and DIPEA (2.0 g, 15.4 mmol). The reaction mixture was stirred at room temperature for 15 h, then diluted with water and extracted with EtOAc. The organic phase was dried over magnesium sulfate, filtered, and evaporated. The residue was purified by silica gel column chromatography (PE / EtOAc, 20% to 100%) to give 3-5 (4.8 g, 6.1 mmol, 79% yield, 98% purity) as a solid. MS: m / z 782 [M−H] - . 1H NMR (DMSO-d6): δ11.31(d,J=2.1Hz,1H),7.82(d,J=8.6Hz,1H),7.63(d,J=8.1Hz,1H),7.50- 7.16(m,9H),6.90(d,J=8.4Hz,4H),5.88(d,J=8.1Hz,1H),5.69(d,J=4.7Hz,1H),5.40(dd,J =8.1,2.0Hz,1H),4.65(q,J=7.9Hz,1H),4.01(d,J=5.8Hz,1H),3.74(s,6H),3.22(m,1H),2. 14(td,J=7.1,4.0Hz,2H),1.49-1.40(m,2H),1.22(d,J=7.6Hz,26H),0.85(t,J=6.6Hz,3H).

[0303] Preparation of 3 To a suspension of 3-5 (2.7 g, 3.4 mmol) in dry DCM (30 mL) was added CN(CH)OP[N(iPr)] (1.3 g, 4.4 mmol) and DCI (341 mg, 2.9 mmol) under a N atmosphere. The mixture was stirred at room temperature for 2.5 h. The solution was then washed with water and brine and dried over NaSO. The combined residue was purified by silica gel column chromatography (PE / EtOAc 5 / 1 to 1 / 3) to give compound 3 (2.4 g, 2.4 mmol, 70% yield) as a white solid. MS: m / z 982 [M−H] - . 1H NMR (DMSO-d6): δ11.41(d,J=4.4Hz,1H),7.99(t,J=9.5Hz,1H),7.66(dd,J=8.2,1.5Hz,1H),7.50-7.13(m,9H) ,6.89(dd,J=9.0,2.4Hz,4H),5.98-5.91(m,1H),5.42(dd,J=8.3,2.6Hz,1H),4.88(td,J=8.5,6.0Hz,1H),4.5 2-4.09(m,2H),3.74(s,7H),3.63-3.44(m,2H),3.28(d,J=3.4Hz,2H),2.75(t,J=6.0Hz,1H),2.66(t,J=6.4Hz ,1H),2.13(ddd,J=17.1,14.2,7.0Hz,2H),1.22(d,J=9.3Hz,27H),1.15-0.91(m,12H),0.85(t,J=6.6Hz,3H). 31 P NMR (DMSO-d6): δ148.88,147.48.

[0304] Example 4.

[0305] [ka]

[0306] Preparation of 4-1 A DMF solution (20 mL) of 3-4 (2.1 g, 3.9 mmol) and hexadecyl isocyanate (1.1 g, 3.9 mmol) was stirred at 30 °C for 2 h. The resulting mixture was diluted with EtOAc, washed with H2O, and dried over Na2SO4. The concentrated residue was purified by silica gel column chromatography (DCM / MeOH 100 / 1 to 10 / 1) to give 4-1 (2.7 g, 3.2 mmol, 82% yield) as a yellow solid. ESI-LCMS: m / z 813 [M+H] + . 1H-NMR (DMSO-d6): δ 11.30 (s, 1H, DO), 7.63 (d, J = 8Hz, 1H), 7.41-7.21 (m, 9H), 6.90 (d, J = 8Hz, 4H), 6.27 (t, J = 5Hz, 1H, exchanged with DO), 5.98 (d, J = 8Hz, 1H, exchanged with DO), 5.86 (d, J = 5Hz, 1H), 5.06 (m, 9H). 78(d,J=8Hz,1H,replaced with D2O),5.41-5.39(m,1H),4.45(dd,J=8Hz,1H),4.09(m,1H),3.98(s, 1H), 3.74 (s, 6H), 3.26-3.15 (m, 2H), 2.98-2.88 (m, 2H), 1.37-1.15 (m, 29H), 0.85 (m, 3H).

[0307] 4. Preparation of HCl To a solution of 4-1 (2.1 g, 2.6 mmol) in DCM (20 mL) was added DCI (313 mg, 2.2 mmol) and CN(CH2)2OP[N(iPr)2] (932 mg, 3.1 mmol) under a N2 atmosphere. The mixture was stirred at 30 °C for 2 h, then diluted with DCM, washed with HO, and dried over Na2SO4. The residue was purified by silica gel column chromatography (PE / EtOAc 2 / 1 to 1 / 2) to give compound 4 (2.3 g, 86% yield) as a white solid. MS: m / z 1013 [M+H] + . 1 H-NMR (DMSO-d6): δ11.41(s,1H),7.65(t,J=8Hz,1H),7.40-7.21(m,9H),6.89(d,J=8Hz,4 H),6.26(dt,J=37.9,5.5Hz,1H),5.92(d,J=9Hz,1H),5.87-5.84(m,1H),5.42(d,J=8Hz,1 H),4.80-4.61(m,1H),4.44-4.13(m,2H),3.90-3.47(m,10H),3.31-3.16(m,2H),3.05-2. 89(m,2H),2.77-2.68(m,2H),1.34-1.19(m,29H),1.15-0.99(m,12H),0.86-0.83(m,3H); 31 P-NMR (DMSO-d6): δ148.94,147.04.

[0308] Example 5.

[0309] [ka]

[0310] Preparation of 5-2 To a solution of (3R,4S,5R)-5-(hydroxymethyl)tetrahydrofuran-2,3,4-triol (100 g, 666 mmol) in pyridine (1000 mL) was added tetraisopropyl 1,3-dichlorodisiloxane at −35° C. under Ar. The reaction mixture was warmed and stirred at room temperature for 16 hours. Acetic anhydride (330 mL) was then added, and the reaction mixture was stirred at room temperature for an additional 6 hours. The reaction mixture was quenched with water (300 mL) at 0° C. and concentrated. The aqueous residue was diluted with EtOAc (1000 mL) and washed with water (2 × 500 mL), 2 N hydrochloric acid (2 × 200 mL), saturated aqueous NaHCO (2 × 400 mL), water (2 × 500 mL), and saturated brine (2 × 400 mL). The organic layer was dried (anhydrous NaSO), filtered, and concentrated under reduced pressure. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (50:1 to 20:1) to give 160 g (50%) of 5-2 as a yellow liquid. MS: m / z 499.30 [M+Na] + .

[0311] Preparation of 5-3 To a solution of 5-2 (160 g, 336 mmol) and p-thiocresol (50 g, 403 mmol, 1.2 equiv.) in anhydrous dichloromethane (1600 mL) was added dropwise a 1 M solution of tin tetrachloride in dichloromethane (74 mL) under an argon atmosphere at 0 °C. The resulting solution was stirred at room temperature for 3 h. The reaction was quenched with trimethylamine (50 mL) and concentrated. The crude residue was purified on a silica gel column using petroleum ether / ethyl acetate (100:1 to 80:1) to give 100 g (55%) of 5-3 as a pale yellow oil. MS: m / z 563.20 [M+Na] + . 1H NMR(CDCl3):δ7.45-7.39(m,2H),7.13(d,J=7.9Hz,2H),5.38(dd,J=5.2,1.5Hz,1H),5.20(d,J=1.5H z,1H),4.06-3.98(m,2H),3.92-3.82(m,2H),2.32(d,J=5.1Hz,3H),2.09(s,3H),1.12-0.95(m,28H).

[0312] Preparation of 5-4 To a solution of 5-3 (80 g, 148 mmol) and 3-azido-1-propanol (37.4 g, 370.2 mmol, 2.5 equiv.) in dichloromethane (800 mL) under Ar, N-iodosuccinimide (36.7 g, 162.9 mmol, 1.1 equiv.) and silver trifluoromethanesulfonate (3.8 g, 14.8 mmol, 0.1 equiv.) were added at -20 °C. The resulting solution was stirred at -20 °C for 30 min, quenched with trimethylamine (30 mL), and concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (50:1 to 20:1) to give 52 g (68% yield) of 5-4 as a yellow oil. MS: 540.40 m / z [M+Na] + . 1 H NMR(CDCl3):δ5.22(d,J=4.8Hz,1H),4.86(s,1H),4.52(dd,J=7.1,4.8Hz,1H),4.05-3.92(m,2H),3.87-3 .70(m,2H),3.49-3.40(m,1H),3.36(t,J=6.7Hz,2H),2.11(s,3H),1.87-1.76(m,2H),1.12-0.96(m,28H).

[0313] Preparation of 5-5 To a solution of 5-4 (34 g, 65.8 mmol) in methanol (340 mL) was added a 5 M solution of NaOMe in MeOH (19.7 mL) under an Ar atmosphere at 0 °C. The resulting solution was stirred at 0 °C for 1 h, then quenched with glacial acetic acid and concentrated. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (40:1 to 10:1) to give 24.5 g (78% yield) of 5-5 as a yellow oil. MS: m / z 498.25 [M+Na] +.

[0314] Preparation of 5-6 To a solution of 5-5 (25 g, 52.6 mmol) in tert-butanol (250 mL) under Ar, acrylonitrile (56.4 g, 1052.6 mmol, 20 equiv.) and cesium carbonate (17.2 g, 52.6 mmol, 1.0 equiv.) were added and stirred at room temperature for 2 h. The resulting mixture was filtered, and the filtrate was concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (30:1 to 8:1) to give 20 g (74% yield) of 5-6 as a yellow oil. MS: 546.40 m / z [M+NH4 + ] + . 1 H NMR(CDCl3):δ4.86(s,1H),4.50(dd,J=8.2,4.0Hz,1H),4.24-4.13(m,1H),4.04-3.88(m,3H),3.84-3.73( m,3H),3.52-3.43(m,1H),3.38(t,J=6.8Hz,2H),2.73-2.56(m,2H),1.89-1.77(m,2H),1.16-1.01(m,28H).

[0315] Preparation of 5-7 To a solution of 5-6 (10 g, 18.9 mmol) in 100 mL of 7 M NH3 in methanol was added Raney Ni (50%, 5 g). The mixture was hydrogenated at 50 °C under 5 atm hydrogen pressure for 24 h. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated to give 7.6 g (77% yield) of 5-7 as a yellow solid, which was used in the next step without further purification. MS m / z [M+H] + (ESI):507.30.

[0316] Preparation of 5-8 To a solution of 5-7 (5.6 g, 11.1 mmol) and palmitic acid (6.2 g, 24.4 mmol, 2.2 equiv.) in dichloromethane (56 mL) was added 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.3 g, 27.6 mmol, 2.5 equiv.), N,N-diisopropylethylamine (7.2 g, 55.3 mmol, 5.0 equiv.), and 1-hydroxybenzotriazole (3.73 g, 27.65 mmol, 2.5 equiv.) at 0 °C under Ar. The reaction mixture was stirred at room temperature for 12 h. The reaction mixture was diluted with dichloromethane (400 mL) and washed with water. The organic layer was dried, filtered, and concentrated. The residue was applied to a silica gel column and eluted with dichloromethane / methanol (100:1 to 80:1) to give 6.0 g (55%) of 5-8 as a white solid. MS: m / z 983.90 [M+H] + . 1 H NMR(CDCl3): δ6.24(d,J=24.1Hz,2H),4.83(s,1H),4.45(dd,J=8.3,4.3Hz,1H),3.98 -3.94(m,3H),3.92-3.85(m,1H),3.78-3.69(m,2H),3.67(d,J=4.3Hz,1H),3.54-3.4 1(m,3H),3.28(dd,J=13.0,6.1Hz,1H),3.21-3.07(m,1H),2.20-2.09(m,4H),1.85-1 .69(m,4H),1.65-1.57(m,4H),1.26(s,48H),1.09-1.02(m,28H),0.90-0.85(m,6H).

[0317] Preparation of 5-9 To a solution of triethylamine trihydrofluoride (2.95 g, 18.3 mmol) in anhydrous THF (60 mL) under Ar, triethylamine (3.69 g, 36.6 mmol) was added and stirred at room temperature for 10 min. A solution of 5-8 (6 g, 6.1 mmol) in anhydrous THF (60 mL) was added, and the reaction mixture was stirred at room temperature for 1 h. It was then diluted with dichloromethane (300 mL) and washed with water and saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The residue was dissolved in 10 mL of dichloromethane and slowly poured into 200 mL of acetonitrile with stirring. The solid was collected by filtration. 3 g (66% yield) of 5-9 was obtained as a white solid. MS: m / z 742.00 [M+H] + .

[0318] Preparation of 5-10 To a solution of 5-9 (2.8 g, 3.78 mmol) in pyridine (60 mL) was added 4,4'-dimethoxytrityl chloride (1.54 g, 1.53 mmol, 1.2 equiv.) under argon at 0 °C and stirred at 25 °C for 4 h. The reaction mixture was quenched with methanol (10 mL), diluted with dichloromethane (200 mL), and washed with saturated aqueous sodium bicarbonate and saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The residue was purified on a silica gel column using dichloromethane / methanol (100:1 to 60:1) to give 3.1 g (79% yield) of 5-10 as a light yellow oil. MS: m / z 1041.75 [M−H] - . 1HNMR(CDCl3):δ7.55-7.41(m,1H),7.37(d,J=1.0Hz,1H),7.34(d,J=1.0Hz,1H),7.32-7.27(m,3H),7.20-7.14(m,3 H),6.87-6.76(m,4H),6.14-5.84(m,2H),5.03-4.93(m,1H),4.34-4.17(m,1H),4.16-4.01(m,2H),3.86-3.81(m,1 H),3.80(s,4H),3.78(s,2H),3.77-3.74(m,1H),3.72-3.61(m,2H),3.58-3.43(m,2H),3.41-3.26(m,3H),3.23-3. 06(m,1H),2.75(s,1H),2.19-2.04(m,4H),1.95-1.70(m,4H),1.69-1.54(m,4H),1.26(s,48H),0.93-0.81(m,6H).

[0319] 5. Preparation To a solution of 5-10 (3.1 g, 3 mmol) in dichloromethane (31 mL) was added 3-(bis(diisopropylamino)phosphinooxy)propanenitrile (1.1 g, 3.6 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (386 mg, 3.3 mmol, 1.1 equiv.) under Ar and stirred at room temperature for 40 min. The resulting solution was diluted with dichloromethane (500 mL) and washed with saturated aqueous sodium bicarbonate and saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The crude product was purified by preparative flash chromatography (C18 column; mobile phase, water and tetrahydrofuran, gradient 30% to 100% THF) to give 2.17 g (60% yield) of compound 5 as a white solid. MS: m / z 1265.95 [M+Na] + . 1HNMR (DMSO-d6): δ7.72-7.65(m,2H),7.47-7.41(m,2H),7.33-7.25(m,6H),7.23-7.16(m,1H),6.87(dd,J=8.6,4.7Hz,4H) ,4.98(d,J=5.8Hz,1H),4.35-4.13(m,1H),4.08-4.01(m,1H),3.77-3.66(m,9H),3.63-3.40(m,6H),3.30-3.17(m,1H),3. 16-3.09(m,2H),3.09-3.00(m,2H),2.94(dd,J=10.3,5.4Hz,1H),2.75(t,J=6.0Hz,1H),2.57-2.53(m,1H),2.06-1.98(m, 4H),1.69-1.55(m,4H),1.52-1.42(m,4H),1.23(s,48H),1.12-1.04(m,9H),0.91(d,J=6.8Hz,3H),0.85(t,J=6.6Hz,6H). 31 PNMR (DMSO-d6): δ148.86,148.57.

[0320] Example 6

[0321] [ka]

[0322] Preparation of 6-2 To a solution of 6-1 (45 g, 94.1 mmol) and 3-azidopropan-1-ol (23.8 g, 235 mmol, 2.5 equiv.) in dichloromethane (450 mL) was added NIS (23.3 g, 103.6 mmol, 1.1 equiv.) and silver trifluoromethanesulfonate (2.4 g, 9.4 mmol, 0.1 equiv.) at -20 °C under Ar. The resulting solution was stirred at -20 °C for 1 h, then quenched with 20 mL of trimethylamine and concentrated. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (100:1 to 10:1) to give 25 g (58% yield) of 6-2 as a yellow oil. MS: m / z 473.30 [M+NH4] + .

[0323] Preparation of 6-3 To a solution of 6-2 (25 g, 54.9 mmol) in MeOH (250 mL) was added a 5 M solution of NaOMe in methanol (13.2 mL) at 0 °C under Ar. The resulting mixture was stirred at 0 °C for 0.5 h, then quenched with glacial acetic acid and concentrated. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (100:1 to 8:1) to give 20 g (88% yield) of compound 6-3 as a yellow oil. MS: m / z 431.30 [M+Na] + .

[0324] Preparation of 6-4 To a solution of 6-3 (20 g, 48.42 mmol) in DMF (200 mL) was added sodium hydride (60% dispersion in oil, 9.68 g, 242 mmol, 5.0 equiv.) at 0 °C and stirred for 30 min at 0 °C. 1-Bromo-2-methoxyethane (13.3 g, 96.9 mmol, 2.0 equiv.) was added, and the resulting solution was stirred at room temperature for 16 h. The reaction mixture was quenched with 200 mL of saturated ammonium chloride solution and then extracted with ethyl acetate (2 × 200 mL). The combined organic layers were washed with water and brine, dried (anhydrous NaSO), filtered, and concentrated. The crude product was applied to a silica gel column and eluted with dichloromethane / methanol (100:1 to 80:1) to give 16 g (70% yield) of 6-4 as a yellow oil. MS: m / z 489.30 [M+Na] + . 1 HNMR(DMSO-d6)δ7.40-7.22(m,10H),4.94(d,J=1.1Hz,1H),4.63-4.42(m,4H),4.10-4.01(m,1H),4.00 -3.92(m,1H),3.90-3.84(m,1H),3.74-3.51(m,4H),3.49-3.35(m,4H),3.25(s,5H),1.74-1.63(m,2H).

[0325] Preparation of 6-5 To a solution of 6-4 (16 g, 34 mmol) in 320 mL of a 10:1 mixture of tetrahydrofuran and water was added triphenylphosphine (17.8 g, 67.9 mmol, 2.0 equiv.). The reaction mixture was stirred at room temperature for 12 h and then concentrated in vacuo. The residue was purified by flash preparative HPLC using a C18 column and a mobile phase of water (containing 0.05% NH4HCO3) and MeCN (gradient 20% to 100% MeCN) to give 13 g (86% yield) of 6-5 as a yellow oil. MS: m / z: 446.25 [M+H] + .

[0326] Preparation of 6-6 To a solution of 6-5 (13 g, 29.2 mmol) and palmitic acid (11.2 g, 43.8 mmol, 1.5 equiv.) in dichloromethane (130 mL) under Ar at 0 °C, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (8.4 g, 43.8 mmol, 1.5 equiv.), N,N-diisopropylethylamine (11.3 g, 87.6 mmol, 3 equiv.), and 1-hydroxybenzotriazole (5.91 g, 43.8 mmol, 1.5 equiv.) were added. The reaction mixture was stirred at room temperature for 12 h, then diluted with dichloromethane (200 mL) and washed with water. The organic layer was dried (anhydrous NaSO), filtered, and concentrated under reduced pressure. The residue was purified on a silica gel column using dichloromethane / methanol (100:1 to 60:1) to give 11 g (55% yield) of 6-6 as a yellow oil. MS m / z 706.45 = [M+Na] + .

[0327] Preparation of 6-7 To a solution of 6-6 (11 g, 16.10 mmol) in MeOH / THF (1:1, 110 mL), Pd / C (10%, 1.1 g) and acetic acid (5 mL) were added and stirred under a H atmosphere for 8 h. The mixture was filtered, and the filtrate was concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (20:1 to 4:1) to give 7.5 g (92% yield) of 6-7 as a white solid. MS: m / z: 526.40 [M+Na] + .

[0328] Preparation of 6-8 To a solution of 6-7 (7.5 g, 14.9 mmol) in pyridine (75 mL) was added 4,4'-dimethoxytrityl chloride (6.04 g, 17.9 mmol, 1.2 equiv.) under argon at 0 °C. The resulting solution was stirred at room temperature for 12 h. The reaction mixture was quenched with MeOH (10 mL), diluted with dichloromethane (10 mL), and washed with saturated aqueous sodium bicarbonate and saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The residue was applied to a silica gel column and eluted with dichloromethane / methanol (100:1 to 40:1) to give 8 g (66% yield) of 6-8 as a pale yellow oil. MS: m / z 804.45 [M−H] - . 1 HNMR(DMSO-d6)δ7.74-7.63(m,1H),7.47-7.40(m,2H),7.34-7.15(m,6H),6.92-6.83(m,4H),4.90(d,J=1.1Hz,1H),4.07-3.87(m,3H),3.79 -3.58(m,9H),3.53-3.35(m,3H),3.26(s,3H),3.17-2.88(m,4H),1.99 (d,J=3.1Hz,3H),1.64-1.36(m,4H),1.23(s,24H),0.95-0.80(m,3H).

[0329] 6. Preparation of HCl To a solution of 6-8 (8 g, 9.9 mmol) in dichloromethane (80 mL) under Ar, 3-((bis(diisopropylamino)phosphaneyl)oxy)propanenitrile (4.2 g, 13.9 mmol, 1.4 equiv.) and 4,5-dicyanoimidazole (1.4 g, 11.9 mmol, 1.2 equiv.) were added. The reaction mixture was stirred at room temperature for 1 h, then diluted with dichloromethane (80 mL) and washed with saturated aqueous sodium bicarbonate and saturated brine. The organic phase was dried (anhydrous Na2SO4), filtered, and concentrated. The residue was purified by flash preparative HPLC (C18 column; mobile phase, water (containing 0.05% NH4HCO3) and THF; gradient 20% to 100% THF) to give 5.07 g (49% yield) of compound 6 as a colorless oil. MS: m / z 1006.85 [M+H]+ . 1 H NMR(DMSO-d6)δ7.66(t,J=5.6Hz,1H),7.51-7.40(m,2H),7.35-7.13(m,7H),6.95-6.81(m,4H),5.01-4.90(m,1H),4 .38-4.11(m,1H),4.03(d,J=6.4Hz,1H),3.81-3.63(m,11H),3.57-3.38(m,6H),3.25(d,J=3.0Hz,3H),3.20-3.13(m, 1H),3.01(d,J=3.2Hz,2H),2.97-2.87(m,1H),2.79-2.70(m,1H),2.60-2.52(m,1H),2.04-1.94(m,2H),1.58(d,J=6. 1Hz,2H), 1.46(d,J=7.5Hz,2H),1.22(d,J=2.2Hz,24H),1.13-1.01(m,9H),0.90(d,J=6.7Hz,3H),0.87-0.81(m,3H). 31 P NMR(DMSO-d6)δ148.92,148.41.

[0330] Example 7

[0331] [ka]

[0332] Preparation of 7-1 To a solution of 6-1 (6 g, 12.5 mmol) in dichloromethane (180 mL) was added MeOH (4 g, 125.3 mmol, 10 equiv.), NIS (3.1 g, 13.8 mmol, 1.1 equiv.), and trimethylsilyl trifluoromethanesulfonate (557 mg, 2.5 mmol, 0.2 equiv.) at -20 °C under N2. The resulting solution was stirred at -20 °C for 1 h and then quenched with aqueous sodium thiosulfate (300 mL). The mixture was extracted with dichloromethane (3 × 600 mL), and the combined organics were dried (anhydrous Na2SO4), filtered, and concentrated. The residue was purified on a silica gel column using ethyl acetate / petroleum ether (1:50 to 1:1) to give 4 g (72% yield) of 7-1 as a yellow oil.

[0333] Preparation of 7-2 To a solution of 7-1 (6 g, 15.5 mmol) in MeOH (100 mL) was added NaOMe (1 g, 18.6 mmol, 1.2 equiv.) under Ar at 0 °C and stirred at 0 °C for 0.5 h. The resulting solution was diluted with dichloromethane (300 mL) and washed with water. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The residue was applied to a silica gel column and eluted with ethyl acetate / petroleum ether (1:10 to 5:1) to give 4.5 g (84%) of 7-2 as a yellow oil. MS: m / z 386.17 [M+H]+.

[0334] Preparation of 7-3 To a solution of 7-2 (4.5 g, 13.1 mmol) in DMF (90 mL) was added sodium hydride (60% in mineral oil, 2.6 g) at 0 °C, and the resulting mixture was stirred for 15 min. 1-Iodohexadecane (9.21 g, 26.1 mmol, 2.0 equiv) was added, and the mixture was stirred at room temperature for 2 h. The reaction mixture was quenched with water (200 mL) and extracted with dichloromethane (3 × 500 mL). The combined organic layers were dried (anhydrous NaSO), filtered, and concentrated. The residue was purified on a silica gel column using ethyl acetate / petroleum ether (1:50 to 1:1) to give 5.5 g (74% yield) of 7-3 as a yellow oil. MS: m / z 568.41 [M+Na] + . 1 H NMR(DMSO-d6)δ7.47-7.20(m,10H),4.83(d,J=1.1Hz,1H),4.60-4.39(m,4H),4.10-4.01(m,1H),3.96-3.90(m,1H) ,3.80-3.73(m,1H),3.60-3.36(m,4H),3.22(s,3H),1.60-1.37(m,2H),1.23(d,J=2.3Hz,27H),0.95-0.77(m,3H).

[0335] Preparation of 7-4 To a solution of 7-3 (5.5 g, 9.7 mmol) in MeOH (550 mL) was added 10% Pd / C (2.25 g), and the resulting mixture was stirred under H for 16 h. The solid was filtered off, and the filtrate was concentrated to give 3 g (80% yield) of 7-4 as a white solid, which was used in the next step without further purification. MS: m / z 388.32 [M+H] + .

[0336] Preparation of 7-5 To a solution of 7-4 (3.2 g, 8.2 mmol) in pyridine (64 mL) under N was added 4,4'-(chloro(phenyl)methylene)bis(methoxybenzene) (2.93 g, 8.6 mmol, 1.1 equiv.) at 0 °C. The resulting mixture was stirred at room temperature for 2 h, then diluted with dichloromethane (200 mL) and washed with water. The organic layer was dried (anhydrous NaSO), filtered, and concentrated. The resulting residue was purified on a silica gel column using ethyl acetate / petroleum ether (1:50 to 1:1) to give 4.1 g (72%) of 7-5 as a yellow oil. MS: m / z 690.45 [M+H] + . 1 H NMR(DMSO-d6)δ7.47-7.38(m,2H),7.33-7.20(m,7H),6.92-6.82(m,4H),4.80(d,J=1.0Hz,1H),4.07-3.86(m,2H),3.74(s,6H),3.63-3.43(m, 3H),3.27(s,3H),3.10(dd,J=10.0,2.6Hz,1H),2.96(dd,J=9.9,5.5Hz,1H),1.50(d,J=6.9Hz,2H),1.25(d,J=8.1Hz,26H),0.87-0.84(m,3H).

[0337] 7. Preparation of Calcium Carbonate To a solution of 7-5 (3 g, 4.3 mmol) in dichloromethane (30 mL), 3-((bis(diisopropylamino)phosphaneyl)oxy)propanenitrile (1.7 g, 5.6 mmol, 1.3 equiv.) and 4,5-dicyanoimidazole (564 mg, 4.8 mmol, 1.1 equiv.) were added, and the resulting solution was stirred at room temperature for 1 h. It was then diluted with dichloromethane and washed with aqueous sodium bicarbonate. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The residue was purified on a silica gel column using ethyl acetate / petroleum ether (1:50 to 1:10) to give 1.9 g (48%) of compound 7 as a light yellow oil. MS: m / z 891.60 [M+H] + . 1 H NMR(DMSO-d6)δ7.49-7.37(m,2H),7.37-7.14(m,7H),6.88(dd,J=8.9,2.9Hz,4H),4.8 7(d,J=2.3Hz,1H),4.36-4.09(m,1H),4.04(d,J=6.3Hz,1H),3.74(d,J=1.4Hz,9H),3. 62-3.44(m,5H),3.25-3.11(m,1H),2.96(dd,J=9.9,5.3Hz,1H),2.80-2.66(m,1H),2. 56(d,J=5.9Hz,1H),1.50(s,2H),1.24(s,27H),1.13-1.03(m,8H),0.95-0.82(m,7H). 31 P NMR(DMSO-d6)δ148.87,148.5.

[0338] Example 8

[0339] [ka]

[0340] Preparation of 8-1 To a solution of 6-1 (72 g, 150.6 mmol) and 1-hexadecanol (44 g, 180.58 mmol, 1.2 equiv.) in dichloromethane (720 mL) under an inert atmosphere of argon, N-iodosuccinimide (37 g, 151.1 mmol, 1.1 equiv.) and silver trifluoromethanesulfonate (4 g, 15.06 mmol, 0.1 equiv.) were added at -20 °C. The resulting solution was stirred at -20 °C for 30 min. The reaction mixture was quenched with 15 mL of trimethylamine and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (50:1 to 20:1) to give 70 g (79%) of 8-1 as a pale yellow oil. MS: m / z 619.45 [M+Na] + . 1 H NMR(CDCl3)δ7.46-7.18(m,10H),5.25(d,J=4.4Hz,1H),5.00(s,1H),4.61-4.5 8(m,2H),4.46(d,J=11.4Hz,1H),4.29-4.22(m,1H),4.17(dd,J=7.5,4.5Hz,1H ),3.76-3.61(m,2H),3.54(dd,J=10.5,6.0Hz,1H),3.38(dt,J=9.4,6.6Hz,1H) ,2.15(s,3H),1.51(q,J=6.7Hz,2H),1.29(d,J=3.8Hz,26H),0.95-0.88(m,4H).

[0341] Preparation of 8-2 A solution of 8-1 (70 g, 117.4 mmol, 1.0 equiv) was dissolved in 700 mL of methanol under an inert atmosphere of argon, and the mixture was added to 35 mL of a 5 M solution of sodium methanolate in methanol at 0 °C. The resulting solution was stirred at 0 °C for 30 min. The resulting mixture was quenched with 40 mL of 3 M hydrochloric acid and concentrated under reduced pressure. The residue was applied and purified on a silica gel column using petroleum ether / ethyl acetate (80:1 to 50:1) to give 51 g (78%) of 8-2 as a pale yellow oil. MS: m / z 577.45 [M+Na] + .

[0342] Preparation of 8-3 To a solution of 8-2 (51 g, 92.1 mmol) in 510 mL of DMF, sodium hydride (11 g, 460 mmol, 5.0 equiv.) was added at 0 °C. The resulting solution was stirred at 0 °C for 30 min. 1-Iodohexadecane (47 g, 138 mmol, 1.5 equiv.) was then added at 0 °C and stirred at room temperature for 16 h. The reaction mixture was quenched with 200 mL of saturated ammonium chloride solution, diluted with 3000 mL of ethyl acetate, and washed with 2 × 1800 mL of water, 1 × 1800 mL of saturated aqueous sodium thiosulfate, saturated aqueous sodium bicarbonate, and saturated aqueous sodium chloride. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (100:1 to 80:1) to give 49 g (76%) of 8-3 as a yellow solid. MS: m / z 796.85 [M+NH4] + .

[0343] Preparation of 8-4 To a solution of 8-3 (49 g, 61.8 mmol) in 392 mL of tetrahydrofuran and 98 mL of methanol under a hydrogen atmosphere, 10% Pd / C (49 g) and acetic acid (50 mL) were added at room temperature and stirred for 12 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 32 g (86% yield) of 8-4 as a white solid, which was used in the next step without further purification. MS: m / z 630.65 [M+NH4] + .

[0344] Preparation of 8-5 To a solution of 8-4 (30 g, 48.9 mmol) in 300 mL of pyridine under an inert atmosphere of argon, 4,4'-dimethoxytrityl chloride (20 g, 58.8 mmol, 1.2 equiv.) was added and stirred at room temperature for 3 h. The reaction mixture was quenched with 150 mL of methanol, diluted with 2500 mL of dichloromethane, and washed with 2 × 2000 mL of saturated aqueous sodium bicarbonate and 2 × 2000 mL of saturated aqueous sodium chloride. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (60:1 to 30:1) containing 0.05% TEA. 39.1 g (88% yield) of 8-5 was obtained as a pale yellow oil. MS: m / z 899.60 [M−H] - . 1 H NMR(CDCl3)δ7.59-7.47(m,2H),7.41(d,J=8.7Hz,3H),7.34-7.26(m,3H),6.85(dd,J=8.8,6.4Hz,4H),5 .02(dd,J=7.2,1.4Hz,1H),4.24-4.14(m,1H),4.14-4.06(m,1H),3.82(d,J=4.7Hz,6H),3.78-3.63(m,3H) ),3.60-3.51(m,1H),3.46-3.35(m,1H),3.30(dd,J=9.9,3.8Hz,1H),3.17(dd,J=9.9,5.6Hz,1H),2.60(d ,J=8.3Hz,1H),1.70-1.60(m,2H),1.52(d,J=13.7Hz,2H),1.28(d,J=8.8Hz,52H),0.91(t,J=6.7Hz,6H).

[0345] 8. Preparation of HCl To a solution of 8-5 (35 g, 38.9 mmol) in 350 mL of dichloromethane was added 3-(bis(diisopropylamino)phosphinooxy)propanenitrile (14 g, 46.7 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (5 g, 42.8 mmol, 1.1 equiv.) under an inert atmosphere of argon at room temperature. The resulting solution was stirred at room temperature for 1 h, diluted with 500 mL of dichloromethane, and washed with 3 × 2000 mL of saturated aqueous sodium bicarbonate and 3 × 2000 mL of saturated aqueous sodium chloride. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with hexane / ethyl acetate (80:1 to 30:1) containing 0.05% TEA to give 21 g (51% yield) of compound 8 as a colorless oil. MS: m / z 1101.80 [M+H] + . 1 H NMR(CDCl3)δ7.52(dd,J=7.9,4.0Hz,2H),7.40(dd,J=8.5,6.0Hz,4H),7.31(s,1H),7.28-7.18(m,2H),6.83(dd ,J=8.5,6.3Hz,4H),5.03(d,J=5.3Hz,1H),4.44-4.18(m,2H),3.81(d,J=3.6Hz,9H),3.62-3.57(m,5H),3.45-3 .27(m,2H),3.10(dd,J=9.9,5.0Hz,1H),2.63(q,J=6.7,6.1Hz,1H),2.36(t,J=6.6Hz,1H),1.62-1.60(m,1H),1 .50(d,J=6.4Hz,3H),1.26(d,J=14.8Hz,52H),1.19-1.13(m,8H),0.99(d,J=6.7Hz,3H),0.91(t,J=6.6Hz,7H). 31 P NMR(CDCl3)δ149.84,149.38.

[0346] Example 9.

[0347] [ka]

[0348] Preparation of 9-1 To a solution of (3R,4R,5R)-5-(acetoxymethyl)tetrahydrofuran-2,3,4-triyl triacetate (100 g, 314.2 mmol) in 1000 mL of acetone was added iodine at 0°C under an inert atmosphere of argon. The resulting solution was stirred at room temperature for 3.5 hours. The reaction mixture was diluted with 2000 mL of ethyl acetate and washed with 2 x 500 mL of saturated aqueous sodium bicarbonate and 2 x 500 mL of saturated aqueous sodium chloride, respectively. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was dissolved in 700 mL of methanol and cooled to 0°C. 54 mL of 5 M sodium methanolate in methanol was then added and stirred at 0°C for 30 minutes under an inert atmosphere of argon. The reaction mixture was quenched with 90 mL of 3 N hydrochloric acid and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with dichloromethane / methanol (120:1 to 40:1) to give 37 g (62%) of 9-1 as a white solid. MS: m / z 189.05 [M−H] - .

[0349] Preparation of 9-2 To a solution of 9-1 (22 g, 115.8 mmol) in anhydrous DMF (220 mL) under an inert atmosphere of argon, sodium hydride (8.34 g, 347.4 mmol, 3.0 equiv.) was slowly added at 0 °C, followed by the dropwise addition of benzyl bromide (49.5 g, 289.5 mmol, 2.5 equiv.) at 0 °C. The resulting solution was stirred at room temperature for 3 h. The reaction mixture was quenched with 20 mL of saturated ammonium chloride solution, diluted with 500 mL of ethyl acetate, and washed with 2 × 150 mL of water and 2 × 150 mL of saturated aqueous sodium chloride solution. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (80:1 to 20:1) to give 35.2 g (82% yield) of 9-2 as a yellow oil. MS: m / z 393.25 [M+Na] + . 1H NMR (DMSO-d6): δ7.30-7.05(m,10H),5.62(d,J=3.7Hz,1H),4.67-4.47(m,2H),4.38(dd,J=12.1,6.1H z,3H),3.94-3.85(m,1H),3.70-3.49(m,2H),3.38(dd,J=11.2,5.1Hz,1H),1.34(s,3H),1.18(s,3H).

[0350] Preparation of 9-3 A solution of 9-2 (30 g, 81.1 mmol) in 150 mL of trifluoroacetic acid and 150 mL of water was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in 300 mL of pyridine and cooled to 0 °C. 60 mL of acetic anhydride was then added and stirred at room temperature for 3 h. The reaction mixture was quenched with 60 mL of water and concentrated under reduced pressure. The residue was diluted with 1000 mL of ethyl acetate and washed with 2 × 500 mL of water, 1 × 200 mL of 2N hydrochloric acid, 1 × 400 mL of saturated aqueous sodium bicarbonate, 2 × 500 mL of water, and 2 × 400 mL of saturated aqueous sodium chloride. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (20:1 to 8:1) to give 21.5 g (64%) of 9-3 as a pale yellow oil. MS: m / z 437.20 [M+Na] + .

[0351] Preparation of 6-1 To a solution of 9-3 (21.5 g, 51.9 mmol) and 9-4 (7.7 g, 62.3 mmol, 1.2 equiv.) in dichloromethane (215 mL) was added 10 mL of 1 M stannous chloride pentahydrate in dichloromethane at 0 °C under an inert atmosphere of argon and stirred at room temperature for 1 h. The reaction mixture was quenched with 50 mL of trimethylamine and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (60:1 to 20:1) to give 16 g (64%) of 6-1 as a pale yellow oil. MS: m / z 496.20 [M+Na] + .

[0352] Preparation of 9-5 To a dichloromethane solution (50 mL) of 6-1 (5 g, 10.5 mmol) and 1-heptadecanol (3.2 g, 12.6 mmol, 1.2 equiv.) under an inert atmosphere of argon, N-iodosuccinimide (2.6 g, 11.5 mmol, 1.1 equiv.) and silver trifluoromethanesulfonate (270 mg, 1.1 mmol, 0.1 equiv.) were added at -20 °C. The resulting solution was stirred at -20 °C for 30 min. The resulting mixture was quenched with 1 mL of trimethylamine and concentrated under reduced pressure. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (50:1 to 20:1) to afford 4.3 g (67% yield) of 9-5 as a light yellow oil. MS: m / z 633.45 [M+Na] + .

[0353] Preparation of 9-6 To a solution of 9-5 (4.30 g, 7.1 mmol) in methanol (43 mL) was added 1.4 mL of 5 M sodium methanolate in methanol at 0 °C under Ar. The resulting solution was stirred at 0 °C for 30 min, and then the reaction was quenched with hydrochloric acid (3 M, 2 mL) and concentrated. The crude product was applied onto a silica gel column and eluted with petroleum ether / EtOAc (80 / 1 to 50 / 1) to give 3.2 g (80%) of 9-6 as a pale yellow oil. MS: m / z 591.45 [M+Na] + . 1 H NMR(DMSO-d6):δ7.36-7.24(m,10H),5.03(s,1H),4.78(s,1H),4.63(d,J=11.9Hz, 1H),4.56-4.46(m,2H),4.43(d,J=11.9Hz,1H),4.11-4.04(m,1H),3.98(d,J=4.4Hz ,1H),3.83(dd,J=7.3,4.4Hz,1H),3.58-3.49(m,2H),3.42(dd,J=10.6,6.4Hz,1H) ,3.31-3.22(m,1H),1.45-1.34(m,2H),1.22(d,J=12.3Hz,28H),0.88-0.82(m,3H).

[0354] Preparation of 9-7 To a solution of 9-6 (4 g, 7.04 mmol) in 40 mL of DMF was added sodium hydride (850 mg, 35.2 mmol, 5.0 equiv.) at 0 °C. The resulting solution was stirred at 0 °C for 30 min. 1-Iodohexadecane (3.7 g, 10.56 mmol, 1.5 equiv.) was then added at 0 °C and stirred at room temperature for 16 h. The reaction mixture was quenched with 10 mL of saturated ammonium chloride solution, diluted with 300 mL of ethyl acetate, and washed with 2 × 150 mL of water, 1 × 150 mL of saturated aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate solutions, and 1 × 150 mL of saturated aqueous sodium chloride solution. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (100:1 to 80:1) to give 4.1 g (74%) of 9-7 as a yellow solid. MS: m / z 810.85 [M+NH4 + ] + . 1 H NMR(CDCl3):δ7.39-7.29(m,10H),5.02(d,J=1.3Hz,1H),4.68-4.53(m,4H),4.36-4.28(m,1H),4.04(dd,J=6.9,4.7Hz,1 H),3.80-3.50(m,6H),3.44-3.34(m,1H),1.70-1.58(m,2H),1.53(t,J=6.8Hz,2H),1.29(s,55H),0.92(t,J=6.8Hz,6H).

[0355] Preparation of 9-8 To a solution of 9-7 (4.1 g, 5.2 mmol) in 32 mL of tetrahydrofuran and 8 mL of methanol was added 10% Pd / C (410 mg) and acetic acid (4 mL) at room temperature under a hydrogen atmosphere. The reaction mixture was stirred for 12 h, then filtered, and the filtrate was concentrated under reduced pressure to give 2.3 g (73%) of 9-8 as a white solid, which was used in the next step without further purification. MS: m / z 630.65 [M+NH4] + .

[0356] Preparation of 9-9 To a solution of 9-8 (2.1 g, 3.4 mmol) in 21 mL of pyridine under an inert atmosphere of argon, 4,4'-dimethoxytrityl chloride (1.58 g, 4.1 mmol, 1.2 equiv.) was added at room temperature. The resulting solution was stirred at 25 °C for 3 h. The reaction mixture was quenched with 10 mL of methanol, diluted with 200 mL of dichloromethane, and washed with 2 × 150 mL of saturated aqueous sodium bicarbonate and 2 × 150 mL of saturated aqueous sodium chloride, respectively. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (60:1 to 30:1) containing 0.05% TEA to give 2.3 g (73% yield) of 9-9 as a pale yellow oil. MS: m / z 913.60 [M−H] - . 1 H NMR(CD2Cl2):δ7.56-7.49(m,2H),7.43-7.37(m,4H),7.37-7.21(m,3H),6.91-6.82(m,4H),5.0 1(d,J=1.4Hz,1H),4.19-4.10(m,1H),4.07-3.96(m,1H),3.82(s,6H),3.78-3.63(m,3H),3.63-3 .51(m,1H),3.46-3.36(m,1H),3.26(dd,J=9.9,3.7Hz,1H),3.13(dd,J=9.9,5.7Hz,1H),2.55(d, J=8.4Hz,1H),1.70-1.61(m,2H),1.56-1.49(m,2H),1.30(d,J=10.9Hz,55H),0.95-0.90(m,6H).

[0357] 9. Preparation of Calcium Carbonate To a solution of 9-9 (2.3 g, 2.5 mmol) in dichloromethane (23 mL) under argon, 3-(bis(diisopropylamino)phosphinooxy)propanenitrile (910 mg, 3.0 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (330 mg, 2.8 mmol, 1.1 equiv.) were added at room temperature and stirred for 1 h. The resulting solution was diluted with 500 mL of dichloromethane and washed with 3 × 300 mL of saturated aqueous sodium bicarbonate and 3 × 300 mL of saturated aqueous sodium chloride. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified on a silica gel column using hexane / ethyl acetate (80:1 to 30:1) containing 0.05% TEA to give 2.04 g (73% yield) of compound 9 as a pale yellow oil. MS: m / z 1115.85 [M+H] + . 1 H NMR(CD2Cl2):δ7.44-7.36(m,2H),7.33-7.24(m,4H),7.23-7.16(m,2H),7.15-7.08(m,1H),6.81-6.54(m, 4H),4.90-4.85(m,1H),4.29-4.05(m,2H),3.89-3.71(m,1H),3.69(d,J=3.1Hz,6H),3.67-3.61(m,2H),3. 59-3.38(m,5H),3.35-3.13(m,2H),3.00-2.88(m,1H),2.59-2.45(m,1H),2.35-2.22(m,1H),1.54-1.46(m ,2H),1.45-1.35(m,2H),1.26-1.14(m,54H),1.08-1.01(m,9H),0.89(d,J=6.8Hz,3H),0.84-0.76(m,6H). 31 P NMR (CD2Cl2): δ149.67,149.39.

[0358] Example 10.

[0359] [ka]

[0360] Preparation of 10-1 To a solution of 9-6 (5 g, 8.8 mmol) in 50 mL of N,N-dimethylformamide, sodium hydride (60% dispersion in oil, 1.7 g, 43.9 mmol, 5.0 equiv.) was added at 0 °C and stirred for 30 min at 0 °C. 1-Bromo-2-methoxyethane (2.4 g, 17.4 mmol, 2.0 equiv.) was then added, and the resulting solution was stirred at room temperature for 16 h. The reaction mixture was quenched with 100 mL of saturated ammonium chloride solution and then extracted with 2 × 200 mL of ethyl acetate. The combined organic layers were washed with 2 × 100 mL of water and 100 mL of brine and dried over anhydrous sodium sulfate. The crude evaporation residue was purified on a silica gel column using dichloromethane / methanol (100 / 1 to 80 / 1) to give 5 g (90% yield) of compound 10-1 as a colorless oil. MS: m / z 649.45 [M+Na] + . 1 H NMR (DMSO-d6): δ7.39-7.23(m,10H),4.92(d,J=1.1Hz,1H),4.63-4.43(m,4H),4.0 4(td,J=6.4,3.5Hz,1H),3.93(dd,J=7.0,4.6Hz,1H),3.85(dd,J=4.6,1.1Hz,1H), 3.72-3.61(m,2H),3.58-3.49(m,2H),3.48-3.40(m,3H),3.30(dt,J=9.5,6.4Hz,1 H),3.25(s,3H),1.41(t,J=6.5Hz,2H),1.21(d,J=10.0Hz,28H),0.89-0.80(m,3H).

[0361] Preparation of 10-2 To a solution of 10-1 (5 g, 7.9 mmol) in methanol (50 mL) and tetrahydrofuran (50 mL) under a hydrogen atmosphere, 10% Pd / C (500 mg) and acetic acid (5 mL, 87.4 mmol) were added. The resulting mixture was stirred at room temperature for 8 h, then filtered, and the filtrate was concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (20 / 1 to 4 / 1) to give 10-2 (2.7 g, 76% yield) as a white solid. MS: m / z 469.30 [M+Na] + . 1H NMR (DMSO-d6): δ4.81(d,J=1.9Hz,1H),3.92(dd,J=6.1,4.7Hz,1H),3.76-3.67(m,2H),3.66-3.56(m,3H),3.50-3.47(m,1H),3.47 -3.45(m,2H),3.44(dd,J=6.7,1.4Hz,1H),3.36-3.27(m,3H),3.26(s,3H),1.45(d,J=6.6Hz,2H),1.24(s,28H),0.91-0.78(m,3H).

[0362] Preparation of 10-3 To a solution of 10-2 (2.7 g, 6.0 mmol) in pyridine (27 mL) was added 4,4'-dimethoxytrityl chloride (2.4 g, 7.2 mmol, 1.2 equiv.) at 0 °C under an inert atmosphere of argon. The resulting solution was stirred at 25 °C for 6 h. The reaction mixture was quenched with 10 mL of methanol, diluted with 100 mL of dichloromethane, and washed with 2 × 100 mL of saturated sodium bicarbonate solution and 2 × 100 mL of saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (40 / 1 to 10 / 1). 3.5 g (77% yield) of 10-3 was obtained as a colorless oil. MS: m / z 747.45 [M−H] - . 1 H NMR (DMSO-d6): δ7.49-7.39(m,2H),7.37-7.14(m,7H),6.94-6.80(m,4H),4.89(d,J=1.1Hz,1H), 4.07-3.88(m,2H),3.73(s,7H),3.69(d,J=4.2Hz,1H),3.68-3.65(m,1H),3.64-3.56(m,2H),3.4 6(ddd,J=5.6,4.2,1.5Hz,2H),3.33(dt,J=9.5,6.5Hz,1H),3.26(s,3H),3.10(dd,J=10.0,2.6Hz ,1H),2.97(dd,J=9.8,5.6Hz,1H),1.48-1.34(m,2H),1.19(d,J=20.9Hz,28H),0.90-0.78(m,3H).

[0363] 10. Preparation of To a solution of 10-3 (2.5 g, 3.3 mmol) in 24 mL of dichloromethane, 3-((bis(diisopropylamino)phosphaneyl)oxy)propanenitrile (1.13 g, 3.96 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (427 mg, 3.63 mmol, 1.1 equiv.) were added at room temperature. The solution was stirred at 25 °C for 45 min. The resulting mixture was diluted with 100 mL of dichloromethane and washed with 2 × 100 mL of saturated sodium bicarbonate solution and 2 × 100 mL of saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The crude product was purified on a silica gel column using hexane / ethyl acetate (20 / 1 to 6 / 1). The product-containing fractions were concentrated to give 2.0 g (63% yield) of compound 10 as a pale yellow oil. MS: m / z 949.60 [M+H] + . 1 H NMR (DMSO-d6): δ7.44(tdd,J=5.0,4.3,3.4,2.0Hz,2H),7.35-7.24(m,6H),7.24-7.17(m,1H), 6.92-6.80(m,4H),5.03-4.92(m,1H),4.35-4.00(m,2H),3.81-3.60(m,11H),3.60-3.34(m,6H) ,3.26(d,J=3.9Hz,3H),3.15(dd,J=10.0,2.4Hz,1H),2.96(dt,J=10.5,5.4Hz,1H),2.79-2.71 (m,1H),2.56(td,J=5.8,2.1Hz,1H),1.52-1.34(m,2H),1.27-1.01(m,37H),0.92-0.82(m,6H). 31 P NMR (DMSO-d6): δ148.67,148.33.

[0364] Example 11

[0365] [ka]

[0366] Preparation of 11-1 To a solution of (4aR,7R,8R,8aS)-6-methoxy-2-phenylhexahydropyrano[3,2-d][1,3]dioxine-7,8-diol (48 g, 170 mmol) in dichloromethane (480 mL) was added tetrabutylammonium hydrogen sulfate (11.5 g, 34.0 mmol, 0.2 equiv.), 1 M NaOH (850 mL), and bromoacetonitrile (61.3 g, 510.7 mmol, 3.0 equiv.) at 0 °C. The resulting mixture was stirred at room temperature for 2 h, then diluted with dichloromethane (400 mL) and washed with water and saturated brine. The organic layer was dried over Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (80 / 1 to 20 / 1) to give 30.1 g (50% yield) of 11-1 as a white solid. MS: m / z 361.20 [M+H] + . 1 H NMR(CDCl3):δ7.53-7.47(m,2H),7.43-7.38(m,3H),5.58(s,1H),4.95(d,J=3.8Hz,1H),4.61-4.45(m,4H) ),4.33(dd,J=9.7,4.1Hz,1H),4.04(t,J=9.1Hz,1H),3.92-3.73(m,2H),3.72-3.61(m,2H),3.49(s,3H).

[0367] Preparation of 11-2 To a solution of 11-1 (20 g, 55.6 mmol) in THF (200 mL) was added LiAlH (4.2 g, 111.1 mmol, 2.0 equiv.) under Ar at 0 °C. The resulting mixture was stirred at 0 °C for 1 h and then quenched with EtOAc. The solid was filtered off. The filtrate was diluted with dichloromethane and washed with water and saturated brine. The organic layer was dried over NaSO, filtered, and concentrated. The residue was purified on a silica gel column using dichloromethane / methanol (50 / 1 to 10 / 1) to give 16.2 g (79% yield) of 11-2 as a white solid. MS: m / z 369.20 [M+H] + .

[0368] Preparation of 11-3 To a solution of 11-2 (10 g, 27.1 mmol) in dichloromethane (200 mL) under Ar at 0 °C, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (11.4 g, 59.6 mmol, 2.2 equiv.), 1H-benzo[d][1,2,3]triazol-1-ol (18.3 g, 135.5 mmol, 5.0 equiv.), N,N-diisopropylethylamine (7.7 g, 59.6 mmol, 2.2 equiv.), and palmitic acid (20.8 g, 81.3 mmol, 3.0 equiv.) were added. The resulting solution was stirred at room temperature for 16 h, diluted with dichloromethane, and washed with water and saturated brine. The organic layer was dried over NaSO, filtered, and concentrated. The residue was purified on a silica gel column using dichloromethane / methanol (80 / 1 to 20 / 1) to give 8.2 g (36% yield) of 11-3 as a white solid. MS: m / z 845.75 [M+H] + .

[0369] Preparation of 11-4 To a solution of 11-3 (8.2 g, 9.70 mmol) in tetrahydrofuran (164 mL) was added 10% Pd / C (800 mg) at room temperature under H2. The mixture was stirred at 30 °C for 12 h, then filtered and concentrated. The crude product was diluted with dichloromethane (200 mL), filtered and concentrated. 11-4 (4.5 g, 61% yield) was obtained as a colorless oil. MS: m / z 757.70 [M+H] + .

[0370] Preparation of 11-5 To a solution of 11-4 (4.5 g, 5.94 mmol) in CHCl (180 mL) was added 4,4'-dimethoxytrityl chloride (2.2 g, 6.53 mmol, 1.10 equiv), EtN (1.7 mL, 12 mmol), and DMAP (146 mg, 1.2 mmol) at room temperature under Ar. The resulting solution was stirred at 50 °C for 4 h and quenched with methanol. The mixture was then diluted with dichloromethane and washed with aqueous NaHCO and saturated brine. The organic layer was dried over anhydrous NaSO, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (60 / 1 to 30 / 1) to give 3.2 g (50%) of 11-5 as a pale yellow oil. MS: m / z 1081.85 [M+Na] + . 1 H NMR(DMSO-d6)δ7.79(t,J=5.7Hz,1H),7.68(t,J=5.6Hz,1H),7.41(d,J=7.7 Hz,2H),7.35-7.21(m,7H),6.88(d,J=8.6Hz,4H),5.18(d,J=6.0Hz,1H),4. 85(d,J=3.3Hz,1H),3.74(s,6H),3.70-3.48(m,5H),3.40(s,3H),3.31-2.9 7(m,9H),2.09-1.98(m,4H),1.46(s,4H),1.23(s,48H),0.90-0.79(m,6H).

[0371] Preparation of 11 To a solution of 11-5 (3 g, 2.83 mmol) in dichloromethane (30 mL) was added 2-cyanoethyl 3-((bis(diisopropylamino)phosphaneyl)oxy)propanenitrile (1.2 g, 3.96 mmol, 1.4 equiv.) and 4,5-dicyanoimidazole (401 mg, 3.39 mmol, 1.2 equiv.) at room temperature under Ar. The reaction mixture was stirred for 1 h, then diluted with dichloromethane and washed with saturated aqueous NaHCO and saturated brine. The organic layer was dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by flash preparative HPLC under the following conditions: column, C18; mobile phase, water (containing 0.05% NHHCO), and tetrahydrofuran (20% tetrahydrofuran up to 100% in 10 min and held at 100% for 5 min). The aqueous layer was separated and extracted with dichloromethane. The organic layers were combined, dried over anhydrous NaSO, filtered, and concentrated to give 1.9413 g (53%) of compound 11 as a white solid. MS: m / z 1259.90 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.89-7.77(m,1H),7.71-7.60(m,1H),7.41(d,J=7.7Hz, 2H),7.34-7.17(m,7H),6.93-6.81(m,4H),4.92(t,J=3.1Hz,1H),3.81-3.34(m,1 9H),3.32-3.11(m,8H),3.01(t,J=9.3Hz,1H),2.68(d,J=11.9Hz,1H),2.16-1.99 (m,4H),1.56-1.40(m,4H),1.23(s,48H),1.09-0.91(m,10H),0.89-0.76(m,8H). 31 P NMR(DMSO-d6)δ149.17,148.15.

[0372] Example 12

[0373] [ka]

[0374] Preparation of 12-1 To a solution of diacetone-D-glucose (80 g, 307.35 mmol) in tetrahydrofuran (800 mL) at 0 °C under N2, sodium hydride (60% dispersion in mineral oil) (18.4 g, 461.53 mmol, 1.5 equiv.) was added. The resulting solution was stirred for 20 min, and iodomethane (65.4 g, 461.53 mmol, 1.5 equiv.) was added at 0 °C and stirred overnight at room temperature. The reaction was quenched with saturated NH4Cl solution at 0 °C and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (30 / 1 to 20 / 1) to give 67.2 g (80% yield) of 12-1 as a yellow oil. MS: m / z 275.14 [M+H] + .

[0375] Preparation of 12-2 A solution of 12-1 (70 g, 255.474 mmol) in 4 M HCl (700 mL) was stirred at room temperature for 1 h and then concentrated. The residue was dissolved in pyridine (550 mL) under N2 at 0 °C, and acetic anhydride (270 mL) was added and stirred at room temperature overnight. The reaction mixture was concentrated and purified on a silica gel column using petroleum ether / ethyl acetate (30 / 1 to 5 / 1) to give 69 g (75% yield) of 12-2 as a yellow solid. MS: m / z 363.15 [M+H] + .

[0376] Preparation of 12-3 To a solution of 12-2 (50 g, 138.12 mmol) in dichloromethane (750 mL) was added 1-hexadecanol (50.1 g, 207.18 mmol, 1.5 equiv.) at 0 °C under N2. The resulting solution was stirred for 20 min, and boron trifluoride etherate (58.8 g, 414.36 mmol, 3 equiv.) was added dropwise and stirred at room temperature for 3 h. The reaction was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layers were washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (30 / 1 to 5 / 1) to give 37.5 g (50% yield) of 12-3 as a yellow solid. MS: m / z 562.50 [M+NH4]+ . 1 H NMR(CDCl3)δ5.13-4.89(m,2H),4.39(d,J=7.9Hz,1H),4.26-4.05(m,3H),3.85(m ,1H),3.65-3.31(m,6H),2.11-2.06(m,8H),1.25(d,J=2.0Hz,28H),0.87(m,3H).

[0377] Preparation of 12-4 A solution of 12-3 (35 g, 64.33 mmol) in methanol (350 mL) was added with sodium methoxide (12.8 g, 321.69 mmol, 5 equiv.) under N2 at 0 °C and stirred at room temperature for 2 h. The pH of the reaction mixture was adjusted to 7 with acetic acid and then concentrated. The residue was purified on a silica gel column using dichloromethane / methanol (100 / 1 to 20 / 1) to give 16 g (60% yield) of 12-4 as a white solid. MS m / z [M+NH4] + (ESI):436.40.

[0378] Preparation of 12-5 To a solution of 12-4 (18 g, 43.06 mmol, 1.0 equiv) in chloroform (180 mL) under N2 at 0 °C, (dimethoxymethyl)benzene (7.85 g, 51.67 mmol, 1.2 equiv) and copper(II) trifluoromethanesulfonate (779 mg, 2.15 mmol, 0.05 equiv) were added and stirred at room temperature for 4 h. The reaction mixture was concentrated and purified on a silica gel column using petroleum ether / ethyl acetate (30 / 1 to 5 / 1) to give 14.1 g (65% yield) of 12-5 as a white solid. MS: m / z 507.50 [M+NH4] + .

[0379] Preparation of 12-6 To a solution of 12-5 (13 g, 25.69 mmol) in tetrahydrofuran (200 mL) at 0 °C under N was added sodium hydride (60% dispersion in mineral oil) (1.5 g, 38.53 mmol, 1.5 equiv.). The resulting solution was stirred at 0 °C for 30 min, 1-iodohexadecane (18 g, 51.38 mmol, 2.0 equiv.) was added, and the reaction was stirred at room temperature for 2 h. The reaction was quenched with saturated NH4Cl solution and extracted with ethyl acetate. The combined organic layers were washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (30 / 1 to 5 / 1) to give 10.7 g (57% yield) of 12-6 as a white solid. MS: m / z 731.60 [M+H] + .

[0380] Preparation of 12-7 To a solution of 12-6 (11 g, 15.06 mmol, 1.0 equiv) in tetrahydrofuran (100 mL) and dichloromethane (100 mL) was added 10% palladium on activated carbon (11 g) at room temperature. After flushing the reaction mixture with H2 for five cycles, the resulting solution was stirred at room temperature for 4 h, then filtered and concentrated. The residue was purified on a silica gel column using ethyl acetate / petroleum ether (1 / 5 to 1 / 3) to give 7.7 g (80% yield) of 12-7 as a white solid. MS: m / z 660.60 [M+NH4] + .

[0381] Preparation of 12-8 To a solution of 12-7 (7.7 g, 11.99 mmol, 1 equiv.) in dichloromethane (100 mL) under N2 at 0 °C, triethylamine (1.8 g, 17.99 mmol, 1.5 equiv.), 4-dimethylaminopyridine (146.3 mg, 1.19 mmol, 0.1 equiv.), and 1-[chloro(4-methoxyphenyl)benzyl]-4-methoxybenzene (4.8 g, 14.39 mmol, 1.2 equiv.) were added. The resulting mixture was stirred at room temperature for 4 h, then quenched with methanol and diluted with dichloromethane. The organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (30 / 1 to 5 / 1, containing 0.5% triethylamine) to give 12-8 in 94% purity. To further increase the purity, the product was recrystallized by dissolving in dichloromethane (1 mL / g), adding dropwise to acetonitrile (20 mL / g), and then filtering to give 6 g (58% yield) of 12-8 as a white solid with 97% purity. MS: m / z 943.55 [M−H] - . 1 H NMR(CDCl3)δ7.50-7.42(m,1H),7.37-7.27(m,6H),7.23-7.17(m,2H),6.89- 6.78(m,4H),4.33-4.30(m,1H),3.90-3.85(m,2H),3.81-3.75(d,J=6.5Hz,6 H),3.66(d,J=3.1Hz,3H),3.64-3.58(m,1H),3.52(m,2H),3.42-3.30(m,2H) ,3.20-3.07(m,2H),1.68-1.53(m,4H),1.28(s,50H),0.90(t,J=6.7Hz,6H).

[0382] Preparation of 12 To a solution of 12-8 (6 g, 6.15 mmol) in dichloromethane (60 mL), N2bis(diisopropylamino)(2-cyanoethoxy)phosphine (2.2 g, 7.38 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (800 mg, 6.77 mmol, 1.1 equiv.) were added at room temperature and stirred for 1 h. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous NaHCO3 and saturated brine. The organic layer was dried over anhydrous Na2OS4, filtered, and concentrated. The residue was purified by flash preparative HPLC using the following conditions: C-18 column, mobile phase, water (containing 0.04% NH4HCO3), and THF, gradient 30–100% THF. The fraction was diluted with dichloromethane, and the organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (30 / 1 to 5 / 1) to give 2.8291 g (39% yield) of compound 12 as a colorless oil. MS: m / z 1145.90 [M+H] + . 1 H-NMR(CDCl3)δ7.46-7.37(m,2H),7.36-7.32(m,4H),7.26-7.19(m,3H),6.82-6.77(m,4H) ,4.34-4.32(m,1H),4.10-3.95(m,1H),3.90-3.80(m,1H),3.77(s,6H),3.62-3.12(m,15H). 2.54-2.23 (m, 2H), 1.80-1.70 (mm, 4), 1.69-1.25 (m, 52H), 1.07-10.2 (m, 10H), 0.90-0.81 (m, 8H). 31 P-NMR(CDCl3)δ150.60,149.83.

[0383] Example 13

[0384] [ka]

[0385] Preparation of 13-1 (3aR,5S,6S,6aR)-5-((R)-2,2-dimethyl-1,3-dioxolan-4-yl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol (80 g, 307.6 mmol) in DMF (800 mL) was added with sodium hydride (60%, 24.6 g, 615.4 mmol, 2.0 equiv.) at 0 °C under N2 and stirred for 15 min. 1-Iodohexadecane (162 g, 461.4 mmol, 1.5 equiv.) was added to the reaction mixture at room temperature and stirred for 2 h. The reaction mixture was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layer was washed with water and saturated brine, then the organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using ethyl acetate / petroleum ether (1 / 70 to 1 / 20) to give 110 g (75% yield) of 13-1 as a white solid. MS: m / z 485.30 [M+H] + .

[0386] Preparation of 13-2 To a solution of 13-1 (110 g, 227.2 mmol) in tetrahydrofuran (1100 mL) was added 6 M HCl (378 mL, 2.27 mol, 10 equiv.) at 0 °C. The resulting solution was stirred at 40 °C for 12 h and then quenched with pyridine (201 mL, 2.5 mol, 11 equiv.). The resulting solution was concentrated and coevaporated with dry pyridine to give 110 g (crude) of 13-2 as a yellow oil, which was used in the next step without further purification. MS: m / z 422.35 [M+NH4] + .

[0387] Preparation of 13-3 To a solution of 13-2 (110 g, crude) in pyridine (1100 mL) was added acetic anhydride (550 mL) at 0 °C under N2 and stirred for 16 h. The reaction mixture was then quenched with water and concentrated. It was then diluted with ethyl acetate and washed with water and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (20 / 1 to 8 / 1) to give 100 g (70% yield for two steps) of 13-3 as a yellow oil. MS: m / z 590.45 [M+NH4] + .

[0388] Preparation of 13-4 To a solution of 13-3 (100 g, 104.8 mmol) and 1-hexadecanol (55.3 g, 157.2 mmol, 1.5 equiv.) in dichloromethane (215 mL) was added 1 M tin(IV) chloride in dichloromethane (314 mL) under Ar at 0 °C and stirred at room temperature for 1 h. The reaction mixture was quenched with trimethylamine and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (60 / 1 to 20 / 1) to give 51 g (64% yield) of compound 13-4 as a pale yellow oil. MS: m / z 772.65 [M+NH4] + . 1 H NMR(CDCl3)δ5.08-4.98(m,2H),4.78(dd,J=10.0,3.7Hz,1H),4.22(dd,J=12.3,4.9Hz,1H) ,4.08(dd,J=12.2,2.3Hz,1H),3.93-3.91(m,J=10.3,5.0,2.3Hz,1H),3.79(t,J=9.6Hz,1H) ,3.72-3.62(m,2H),3.56-3.52(m,1H),3.44(dt,J=10.0,6.6Hz,1H),2.11(d,J=7.5Hz,9H), 1.61(t,J=6.9Hz,2H),1.48(d,J=6.5Hz,2H),1.27(d,J=5.2Hz,52H),0.90(t,J=6.7Hz,6H).

[0389] Preparation of 13-5 To a solution of 13-4 (50 g, 66.26 mmol) in methanol (500 mL) was added 5 M sodium methanolate in methanol (20 mL) at 0 °C under Ar and stirred at room temperature for 2 h. The resulting mixture was quenched with 3 M hydrochloric acid (20 mL) and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (50 / 1 to 1 / 1) to give 25 g (80% yield) of 13-5 as a pale yellow oil. MS: m / z 646.60 [M+NH4] + .

[0390] Preparation of 13-6 To a solution of 13-5 (25 g, 39.80 mmol) in chloroform (250 mL) at 0 °C under N2, benzaldehyde dimethyl acetal (9 g, 59.71 mmol, 1.5 equiv.) and copper(II) trifluoromethanesulfonate (2.8 g, 7.96 mmol, 0.2 equiv.) were added and stirred at room temperature for 16 h. The reaction mixture was quenched with trimethylamine and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (20 / 1 to 10 / 1) to give 17 g (65%) of 13-6 as a yellow oil. MS: m / z 717.65 [M+H] + . 1 H NMR(CDCl3)δ7.52-7.50(m,2H),7.43-7.35(m,3H),5.57(s,1H),4.91(d,J =2.7Hz,1H),4.29(dd,J=10.0,4.6Hz,1H),3.93-3.81(m,2H),3.79-3.70(m ,3H),3.68-3.62(m,2H),3.59-3.46(m,2H),2.36(s,1H),2.32-2.09(m,1H ),1.64(dt,J=15.4,7.4Hz,4H),1.27(d,J=8.9Hz,52H),0.93-0.88(m,6H).

[0391] Preparation of 13-7 To a solution of 13-6 (17 g, 23.74 mmol) in DMF (170 mL) was added sodium hydride (60%, 1.9 g, 47.48 mmol, 2.0 equiv.) at 0 °C under N2 and stirred for 15 min. Iodomethane (5 g, 35.61 mmol, 1.5 equiv.) was added to the reaction mixture and stirred at room temperature for 2 h. The reaction mixture was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layers were washed with water and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (20 / 1 to 8 / 1) to give 12 g (77% yield) of 13-7 as a yellow oil. MS: m / z 731.55 [M+H] + .

[0392] Preparation of 13-8 To a solution of 13-7 (12 g, 23.28 mmol) in tetrahydrofuran (120 mL) at room temperature was added 10% Pd / C (12 g) and flushed with H for five cycles. The resulting solution was stirred at room temperature for 16 h, then filtered and concentrated to give 9 g (80% yield) of 13-8 as a white solid. MS: m / z 660.65 [M+NH] + .

[0393] Preparation of 13-9 To a solution of 13-8 (9 g, 13.99 mmol) in CHCl (500 mL) at 0 °C under Ar, 4,4'-dimethoxytrityl chloride (5.6 g, 16.8 mmol, 1.2 equiv), EtN (3.9 mL, 28 mmol), and DMAP (340 mg, 2.8 mmol) were added and stirred at room temperature for 4 h at 50 °C. The reaction mixture was quenched with methanol, diluted with dichloromethane, and washed with saturated NaHCO and saturated brine. The organic layer was dried over anhydrous NaSO, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (40 / 1 to 2 / 1) to give 6 g (75% yield) of 13-9 as a colorless oil. MS: m / z 943.65 [M−H] - . 1H NMR(CDCl3)δ7.49-7.43(m,2H),7.40-7.32(m,4H),7.32-7.27(m,2H),7.24-7.17(m,1H),6.88-6.77(m,4H),4.97(d,J=3.6Hz,1H),3.92-3 .62(m,10H),3.57-3.46(m,6H),3.38-3.20(m,3H),2.47(d,J=1.9Hz,1H),1.68-1.60(m,4H),1.26(d,J=2.6Hz,52H),0.90(d,J=6.4Hz,6H).

[0394] Preparation of 13 To a solution of 13-9 (6 g, 6.35 mmol) in dichloromethane (60 mL) under Ar at room temperature, 3-(bis(diisopropylamino)phosphinooxy)propanenitrile (2.3 g, 7.62 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (824 mg, 6.98 mmol, 1.1 equiv.) were added and stirred for 1 h. The resulting solution was diluted with dichloromethane and washed with saturated aqueous NaHCO3 and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by preparative flash chromatography using the following conditions: C-18 column, mobile phase, water and tetrahydrofuran, gradient 30% to 100% THF. The product-containing fractions were diluted with an equal volume of dichloromethane, and the aqueous layer was separated and extracted with dichloromethane. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (10 / 1 to 2 / 1) to give 2.0 g (30% yield) of compound 13 as a colorless oil. MS: m / z 1145.90 [M+H] + . 1H NMR(CDCl3)δ7.46(d,J=7.6Hz,2H),7.37-7.33(m,4H),7.29(s,1H),7.25-7.15(m,2H),6.80(dd,J =8.9,3.8Hz,4H),5.01(d,J=3.6Hz,1H),3.92(t,J=9.1Hz,2H),3.78(s,6H),3.76-3.53(m,6H),3. 50(s,4H),3.39-3.18(m,4H),3.08(q,J=10.1,9.0Hz,1H),2.58-2.20(m,2H),1.76(q,J=7.5Hz,2H ),1.42(d,J=6.7Hz,4H),1.33-1.17(m,50H),1.03(dd,J=8.9,6.6Hz,10H),0.87(q,J=5.9Hz,8H). 31 P NMR (DMSO-d6): δ150.69,149.57.

[0395] Example 14.

[0396] [ka]

[0397] Preparation of 14-1 To a solution of (4aR,7R,8R,8aS)-6-methoxy-2-phenylhexahydropyrano[3,2-d][1,3]dioxine-7,8-diol (5 g, 17.73 mmol) in DMF (50 mL) was added sodium hydride (60%, 3 g, 70.92 mmol, 4.0 equiv.) at 0 °C under N2 and stirred for 15 min. 1-Iodohexadecane (12 g, 35.46 mmol, 2.0 equiv.) was added to the reaction mixture and stirred at room temperature for 2 h. The reaction mixture was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layers were washed with water and saturated brine, then dried over anhydrous Na2SO4, filtered, and concentrated. The resulting residue was purified on a silica gel column using ethyl acetate / petroleum ether (1 / 50 to 1 / 1) to give 5 g (50% yield) of 14-1 as a white solid. MS: m / z 731.60 [M+H] + .

[0398] Preparation of 14-2 A solution of 14-1 (5 g, 6.84 mmol, 1.0 equiv.) in tetrahydrofuran (50 mL) was added to 10% palladium-on-activated carbon (w t / w t =100%, 5 g) was added and flushed with H for 5 cycles. The resulting solution was stirred at room temperature for 16 h, then filtered and concentrated to give 3.5 g (80% yield) of 14-2 as a white solid. MS: m / z 643.6 [M+H] + .

[0399] Preparation of 14-3 To a solution of 14-2 (3.5 g, 5.45 mmol, 1 equiv.) in pyridine (40 mL) under N2 at 0 °C, 4,4'-(chloro(phenyl)methylene)bis(methoxybenzene) (2.3 g, 7.08 mmol, 1.3 equiv.) was added and stirred at room temperature for 2 h. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous NaHCO3 and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using ethyl acetate / petroleum ether (1 / 50 to 2 / 1) to give 4 g (78% yield) of 14-3 as a white solid. MS: m / z 943.70 [M−H] - . 1 H NMR(CDCl3)δ7.39-7.32(m,2H),7.29-7.22(m,3H),7.22-7.11(m,3H),7.10-7.04(m, 1H),6.73(dd,J=9.0,2.6Hz,4H),4.70(t,J=3.7Hz,1H),3.86-3.71(m,1H),3.69(d,J= 4.1Hz,6H),3.63-3.37(m,6H),3.33(d,J=3.3Hz,3H),3.30-3.17(m,3H),2.36(dd,J=6 .8,1.8Hz,1H),1.49(q,J=7.5,6.4Hz,4H),1.15(d,J=2.9Hz,52H),0.86-0.71(m,6H).

[0400] Preparation of 14 To a solution of 14-3 (4 g, 4.23 mmol, 1.0 equiv.) in dichloromethane (40 mL) under Ar at 0 °C, 3-(bis(diisopropylamino)phosphinooxy)propanenitrile (1.5 g, 5.07 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (549 mg, 4.65 mmol, 1.1 equiv.) were added and stirred at room temperature for 1 h. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous NaHCO and saturated brine. The organic layer was dried over anhydrous NaSO, filtered, and concentrated. The residue was purified by preparative flash chromatography using the following conditions: column, C18 silica gel; mobile phase, water, and tetrahydrofuran (30% tetrahydrofuran up to 100% in 15 min and hold at 100% for 5 min); detector, UV 254 nm. The residue was diluted with an equal volume of dichloromethane. The organic layer was separated, and the aqueous layer was extracted with dichloromethane. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (10 / 1 to 2 / 1) to give 2.2169 g (50% yield) of compound 14 as a colorless oil. MS: m / z 1145.85 [M+H] + . 1 H NMR(CDCl3)δ7.52-7.46(m,2H),7.40-7.36(m,4H),7.32-7.29(m,1H),7.27(d,J=4.8Hz,1H ),7.26-7.19(m,1H),6.88-6.80(m,4H),4.89(d,J=3.6Hz,1H),3.91-3.87(m,1H),3.92-3.8 7(d,J=1.5Hz,7H),3.73-3.50(m,10H),3.43-3.09(m,5H),2.58-2.20(m,2H),1.63(d,J=6. 7Hz, 2H), 1.52 (d, J=6.5Hz, 2H), 1.28-1.26 (m, 52H), 1.10-1.01 (m, 10H), 0.95-0.83 (m, 8H). 31 P NMR (DMSO-d6): δ150.61,149.21.

[0401] Example 15.

[0402] [ka]

[0403] Preparation of 15-1 To a solution of 5-7 (2.5 g, 5.1 mmol), docosanoic acid (4.3 g, 12.7 mmol, 2.5 equiv.), and N,N-diisopropylethylamine (3.9 g, 30.4 mmol, 6 equiv.) in N,N-dimethylformamide (25 mL) was added O-benzotriazole-N,N,N',N'-tetramethyl-uronium-hexafluorophosphate (5.8 g, 15.2 mmol, 3 equiv.) and 1-hydroxybenzotriazole (1.7 g, 12.7 mmol, 2.5 equiv.) at 0 °C. The resulting solution was stirred at room temperature for 12 h and then diluted with dichloromethane. The organic layer was washed with water and saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using dichloromethane / methanol (100 / 1 to 40 / 1) to give 3 g (52% yield) of 15-1 as a white solid. MS: m / z 1192.05 [M+Na+HO] + .

[0404] Preparation of 15-2 To a solution of triethylamine trihydrofluoride (3.6 g, 22.6 mmol, 10 equiv.) in tetrahydrofuran (52 mL) at room temperature were added triethylamine (4.6 g, 45.1 mmol, 20 equiv.) and 15-1 (2.6 g, 2.3 mmol, 1 equiv.). The solution was stirred at room temperature for 12 h, then precipitated with methanol / water (10:1) and filtered. The solid was washed with acetonitrile to give 1.6 g (78% yield) of 15-2 as a white solid. MS: m / z 909.82 [M+H] + .

[0405] Preparation of 15-3 To a solution of 15-2 (1 g, 1.1 mmol) in dichloromethane (100 mL) at 0 °C under N2, 4,4'-(chloro(phenyl)methylene)bis(methoxybenzene) (447 mg, 1.3 mmol, 1.2 equiv), trimethylamine (222 mg, 2.2 mmol, 2 equiv), and 4-dimethylaminopyridine (13 mg, 0.1 mmol, 0.1 equiv) were added and stirred at 50 °C for 1 h. The reaction mixture was precipitated with acetonitrile and filtered. The residue was purified on a silica gel column using dichloromethane / methanol (100 / 1 to 20 / 1) to give 692 mg (52% yield) of 15-3 as a white solid. MS: m / z 1209.85 [M−H] - .

[0406] Preparation of 15 To a solution of 15-3 (2.8 g, 2.3 mmol) in dichloromethane (56 mL), 3-((bis(diisopropylamino)phosphaneyl)oxy)propanenitrile (0.9 g, 3.0 mmol, 1.3 equiv.) and 4,5-4,5-dicyanoimidazole (300 mg, 2.5 mmol, 1.1 equiv.) were added and stirred at room temperature for 45 min. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous NaHCO3 and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by C18 flash chromatography (mobile phase, THF (0.05% ammonium bicarbonate) in water, 20% to 100% gradient over 25 min). The residue was diluted with an equal volume of dichloromethane. The organic layer was separated, and the aqueous layer was extracted with dichloromethane. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using n-hexane / ethyl acetate (100 / 1 to 1 / 1) to give 2.09 g (61% yield) of compound 15 as a white solid. MS: m / z 1412.05 [M+H] + . 1H-NMR(CDCl3): δ7.49-7.47(m,2H),7.38-7.34(m,4H),7.26-7.21(m,4H),6.81(t,J=6Hz,4H),5.66-5 .49(m,1H),5.00(s,1H),4.41-4.19(m,1H),4.09(s,1H),3.78(s,9H),3.71-3.66(m,1H),3.69-3.34( m,8H),3.19-3.03(m,2H),2.62(t,J=6Hz,1H),2.35(t,J=6Hz,1H),2.17-1.90(m,4H),1.82-1.68(m,4 H),1.63-1.45(m,4H),1.25(s,72H),1.14(t,J=7.5Hz,9H),0.98(d,J=6Hz,3H),0.88(t,J=7.5Hz,6H); 31 P-NMR (CD2Cl2): δ149.81,149.51.

[0407] Example 16.

[0408] [ka]

[0409] Preparation of 16-1 To a solution of methyl tert-butoxycarbonylglycinate (22 g, 116.40 mmol) in 220 mL of N,N-dimethylformamide was added sodium hydride (5 g, 127.5 mmol, 1.1 equiv.) at 0 °C. The resulting solution was stirred at 0 °C for 30 min. 1-Iodohexadecane (49 g, 139.58 mmol, 1.2 equiv.) was then added at 0 °C and stirred at room temperature for 16 h. The reaction mixture was quenched with 50 mL of saturated ammonium chloride solution, diluted with 1500 mL of ethyl acetate, and washed with 2 × 300 mL of water, 1 × 300 mL of saturated aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate solutions, and 1 × 300 mL of saturated brine. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (70 / 1 to 20 / 1) to give 22 g (52% yield) of 16-1 as a yellow solid. MS: m / z 436.35 [M+Na] + .

[0410] Preparation of 16-2 To a solution of 16-1 (22 g, 53.14 mmol) in 110 mL of dichloromethane was added 110 mL of trifluoroacetic acid at 0 °C. The resulting solution was stirred at 0 °C for 1 h. The solvent was removed under reduced pressure, and the crude product was purified on a silica gel column using hexane / ethyl acetate (70 / 1 to 1 / 2) to give 13 g (77% yield) of 16-2 as a white solid. MS: m / z 314.30 [M+H] + .

[0411] Preparation of 16-3 To a solution of 16-2 (13 g, 42.17 mmol) and N,N-diisopropylethylamine (16 g, 122.40 mmol, 3.0 equiv.) in 130 mL of dichloromethane under an inert atmosphere of argon, palmitic acid (13 g, 48.86 mmol, 1.2 equiv.) was added at 0 °C. The mixture was stirred for 10 min, and then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (12 g, 61.15 mmol, 1.5 equiv.) and 1-hydroxybenzotriazole (9 g, 60.79 mmol, 1.5 equiv.) were added at 0 °C. The resulting solution was stirred at room temperature for 16 h, then diluted with 1000 mL of dichloromethane and washed with 2 × 300 mL of water and 2 × 300 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified on a silica gel column using hexane / ethyl acetate (70 / 1 to 2 / 1) to give 13 g (58% yield) of compound 16-3 as a white solid. MS: m / z 552.60 [M+H] + . 1 H NMR(CDCl3)δ4.05(d,J=8.5Hz,2H),3.76(d,J=18.8Hz,3H),3.40-3.29(m,2H),2 .40-2.34(m,2H),1.71-1.53(m,4H),1.35-1.25(m,50H),0.89(d,J=7.0Hz,6H).

[0412] Preparation of 16-4 To a solution of 16-3 (13 g, 23.91 mmol) in tetrahydrofuran (130 mL) was added 5 M lithium hydroxide (24 mL, 119.55 mmol, 5.0 equiv.) at 0 °C under an inert atmosphere of argon. The resulting solution was stirred at room temperature for 3 h. The mixture was diluted with 1000 mL of dichloromethane, adjusted to pH 6 with 2 M hydrochloric acid, and then washed with 2 × 300 mL of water and 2 × 300 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was applied to a silica gel column using hexane / ethyl acetate (70 / 1 to 1 / 2) to give 11 g (83% yield) of 16-4 as a white solid. MS: m / z 538.55 [M+H] + .

[0413] Preparation of 16-5 To a solution of 16-4 (11 g, 19.89 mmol) and N,N-diisopropylethylamine (10 g, 79.6 mmol, 4.0 equiv.) in 100 mL of dichloromethane was added tert-butylmethylglycinate (4 g, 23.6 mmol, 1.2 equiv.) at 0 °C under an inert atmosphere of argon. The mixture was stirred for 10 min, and then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (6 g, 29.7 mmol, 1.5 equiv.) and 1-hydroxybenzotriazole (4 g, 29.5 mmol, 1.5 equiv.) were added at 0 °C. The resulting solution was stirred at room temperature for 12 h, then diluted with 1000 mL of dichloromethane and washed with 2 × 300 mL of water and 2 × 300 mL of saturated brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was applied onto a silica gel column and purified with hexane / ethyl acetate (70 / 1 to 2 / 1) to give 8 g (57% yield) of 16-5 as a white solid. MS: m / z 665.65 [M+H] + .

[0414] Preparation of 16-6 To a solution of 16-5 (8 g, 7.04 mmol) in 50 mL of dichloromethane was added 25 mL of trifluoroacetic acid at 0 °C. The resulting solution was stirred at 0 °C for 2 h. The solvent was removed under reduced pressure, and the crude product was purified on a silica gel column using hexane / ethyl acetate (70 / 1 to 1 / 2) to give 5 g (70% yield) of 16-6 as a white solid. MS: m / z 609.55 [M+H] + .

[0415] Preparation of 16 To a solution of 16-6 (4 g, 6.25 mmol, 1.0 equiv.) in 38 mL of DMF under an inert atmosphere of argon, 1-hydroxypyrrolidine-2,5-dione (2 g, 18.72 mmol, 3.0 equiv.) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (2 g, 9.36 mmol, 1.5 equiv.) were added at 0° C. The reaction mixture was stirred at 25° C. for 16 hours and then cooled to 0° C. The resulting solution was filtered, and the filter cake was washed with cold DMF and cold acetonitrile. The solid was dissolved in dichloromethane and then concentrated to give 1.9 g (43% yield) of compound 16 as a white solid. MS: m / z 706.70 [M+H] + . 1 H NMR(CDCl3)δ4.56(d,J=35.8Hz,2H),4.15(d,J=42.3Hz,2H),3.36(t,J=7.6Hz,2H),3.14(s,2H),3.04(s,1H) ),2.85(d,J=9.7Hz,4H),2.36(t,J=7.6Hz,2H),1.64(d,J=6.8Hz,6H),1.26(s,48H),0.88(t,J=6.7Hz,6H).

[0416] Example 17.

[0417] [ka]

[0418] Preparation of 17-1 To a solution of tetradecane-1,14-diol (54 g, 234.388 mmol) in 540 mL of cyclohexane, hydrogen bromide (20.9 g, 257.827 mmol, 1.1 equiv.) was added at room temperature, and the solution was stirred at 80 °C for 16 h. The reaction was quenched by the addition of saturated sodium bicarbonate solution (500 mL), and the resulting solution was extracted with 3 × 800 mL of dichloromethane. The combined organic layers were washed with 2 × 500 mL of water and 500 mL of brine. The organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (100 / 1 to 4 / 1) to give 42 g (61% yield) of 17-1 as a white solid.

[0419] Preparation of 17-2 To a solution of 17-1 (22 g, 75.0 mmol) in 440 mL of dimethylformamide, sodium hydride (3.6 g, 150.02 mmol, 2.0 equiv.) was added at 0 °C. The solution was stirred at 0 °C for 30 min, and then (bromomethyl)benzene (19.2 g, 112.5 mmol, 1.5 equiv.) was added at 0 °C. The solution was stirred at room temperature for 18 h. The reaction was quenched by the addition of saturated ammonium chloride solution (500 mL), and the resulting solution was extracted with 3 × 500 mL of dichloromethane. The combined organic layers were washed with 2 × 300 mL of water and 300 mL of saturated brine. The organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 4 / 1) to give 21.0 g (73% yield) of 17-2 as a yellow oil.

[0420] Preparation of 17-3 To a solution of 17-2 (50 g, 130.4 mmol) in 400 mL of acetonitrile was added triphenylphosphane (68.4 g, 260.8 mmol, 2.0 equiv.) at room temperature, and the resulting mixture was stirred at reflux temperature for 16 h. The product was precipitated with n-hexane and filtered. The yield of 17-3 as a white solid was 64.0 g (76%). MS: m / z 565 [M+H] + .

[0421] Preparation of 17-4 To a solution of oxalyl chloride (572 mg, 4.51 mmol, 1.5 equiv.) in dry tetrahydrofuran (5 mL) under an inert atmosphere, a solution of dimethyl sulfoxide (704 mg, 9.02 mmol, 3.0 equiv.) in tetrahydrofuran (3 mL) was added dropwise with stirring at −80° C. After 30 min, ((3r,5r,7r)-adamantan-1-yl)methanol (500.0 mg, 3.007 mmol, 1.0 equiv.) dissolved in tetrahydrofuran (3 mL) was added dropwise with stirring at −80° C. After 2 h, triethylamine (1.9 mL, 13.5 mmol, 4.5 equiv.) was added, and stirring was continued at −80° C. for 30 min. The mixture was warmed to room temperature and diluted with dichloromethane (20 mL). The organic layer was washed successively with 2 × 20 mL of saturated ammonium chloride solution and 20 mL of brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 50 / 1) to give 450 mg (88% yield) of 17-4 as a white solid.

[0422] Preparation of 17-5 To a solution of 17-3 (50.0 g, 77.43 mmol) in 500 mL of tetrahydrofuran, sodium hydride (7.4 g, 309.7 mmol, 4.0 equiv.) was added at 0 °C, and the resulting mixture was stirred at 0 °C for 30 min. Then, 17-4 (10.2 g, 61.9 mmol, 0.8 equiv.) was added at 0 °C, and the mixture was stirred at room temperature for 16 h. The reaction was quenched by the addition of saturated ammonium chloride solution (1000 mL) and extracted with 3 × 1500 mL of dichloromethane. The combined organic layers were washed with 2 × 1000 mL of water and 1000 mL of brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (100 / 1 to 3 / 1) to give 31.0 g (88% yield) of 17-5 as a yellow oil. 1H-NMR(CDCl3): δ7.30-7.12(m,5H),5.11-4.89(m,2H),4.41(s,2H),3.37(t,J=6.6Hz,2H),2.09(q,J=6.8Hz, 2H),1.85(dd,J=6.1,3.1Hz,3H),1.71-1.56(m,11H),1.50(dt,J=13.5,6.8Hz,2H),1.19(d,J=12.7Hz,21H).

[0423] Preparation of 17-6 To a solution of 17-5 (12.5 g, 27.7 mmol) in 250 mL of ethyl acetate, palladium on carbon (25 g) was added, and the resulting mixture was stirred at 65 °C under 20 atm of hydrogen for 8 h. The mixture was then filtered, the filtrate was concentrated, and the residue was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 3 / 1). 17-6 (5.25 g, 52% yield) was obtained as a white solid. 1 H-NMR(300MHz,DMSO-d6)δ4.21-4.13(m,1H),3.24(td,J=6.4,5.0Hz,2H),1.79(s,3H),1 .51(dt,J=15.1,10.8Hz,7H),1.30(d,J=2.9Hz,9H),1.12(s,24H),0.89(d,J=7.3Hz,2H).

[0424] Preparation of 17-7 To a solution of 17-6 (3.5 g, 9.7 mmol) in 70 mL of acetone, Jones reagent (11.2 mL) was added at 0 °C, and the resulting mixture was stirred at 40 °C for 24 h. The mixture was then filtered, and the filtrate was precipitated with acetonitrile to give 2.7 g (74% yield) of 17-7 as a white solid. 1 H-NMR (DMSO-d6): δ11.95(s,1H),2.18(t,J=7.3Hz,2H),1.91(s,3H),1.62(q,J=12.3H z,6H),1.52-1.33(m,8H),1.24(s,18H),1.18(d,J=9.2Hz,4H),1.01(d,J=7.2Hz,2H).

[0425] Preparation of 17 To a solution of 17-7 (3 g, 8 mmol) in DMF (60 mL) was added 1-hydroxypyrrolidine-2,5-dione (1.8 g, 15.9 mmol, 2.0 equiv), 4-dimethylaminopyridine (194 mg, 1.6 mmol, 0.2 equiv), N,N-diisopropylethylamine (3.1 g, 23.9 mmol, 3.0 equiv), and 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (2.3 g, 11.94 mmol, 1.5 equiv) at 0 °C. The resulting solution was stirred at room temperature for 12 h and then extracted with 3 × 500 mL of dichloromethane. The combined organic layer was washed with 2 × 500 mL of water and 500 mL of saturated brine. The organic layer was dried (anhydrous NaSO), filtered, and concentrated. The crude residue was applied to a silica gel column and eluted with n-hexane / ethyl acetate (100 / 1 to 3 / 1) to give 2.1 g (53.3% yield) of compound 17 as a white solid. MS: m / z 491 [M+NH4] + . 1 H-NMR:(CDCl3):δ2.83(s,4H),2.62-2.57(t,J=7.5Hz,2H),1.92(s,3H),1.79-1.59( m,8H),1.45-1.35(m,8H),1.33-1.24(m,16H),1.23-1.18(m,4H),1.04-0.96(m,2H).

[0426] Example 18.

[0427] [ka]

[0428] Preparation of 18-1 To a solution of methyl trans-4-aminocyclohexanecarboxylate hydrochloride (5 g, 25.9 mmol, 1.2 equiv.) and N,N-diisopropylethylamine (6.68 g, 51.8 mmol, 2.4 equiv.) in 90 mL of dichloromethane under an inert atmosphere of argon, heptadecanoic acid (5.8 g, 21.6 mmol, 1.0 equiv.) was added at 0 °C, and the mixture was stirred for 10 minutes. N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (4.97 g, 25.9 mmol, 1.2 equiv.) and 1-hydroxybenzotriazole (3.5 g, 25.9 mmol, 1.2 equiv.) were then added at 0 °C. The resulting solution was stirred at room temperature for 12 hours and then diluted with 200 mL of dichloromethane. The organic layer was washed with 2 × 100 mL of water and 2 × 100 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (70 / 1 to 2 / 1) to give 7.1 g (79% yield) of 18-1 as a white solid. MS: m / z 410.50 [M+H] + . 1 H NMR(CDCl3)δ5.35(s,1H),3.92-3.78(m,1H),3.59(s,3H),2.41(s,1H),2.11-2.0 3(m,2H),1.86-1.69(m,2H),1.69-1.39(m,8H),1.15(s,26H),0.81-0.74(m,3H).

[0429] Preparation of 18-2 To a solution of 18-1 (7.1 g, 17.4 mmol) in 71 mL of tetrahydrofuran under an inert atmosphere of argon, lithium hydroxide (5 M, 43.4 mL, 87 mmol, 5.0 equiv.) was added at 0 °C. The resulting solution was stirred at room temperature for 16 h and then neutralized to pH 6 with hydrochloric acid (2 M). The mixture was diluted with 200 mL of dichloromethane and then washed with 2 × 100 mL of water and 2 × 100 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The crude residue was purified on a silica gel column using hexane / ethyl acetate (70 / 1 to 1 / 2) to give 5 g (72% yield) of 18-2 as a white solid. MS: m / z 396.40 [M+H] + .1 H NMR(CDCl3):δ5.29(s,1H),3.85(s,1H),2.48(s,1H),2.05(t,J=7.6Hz,2H),1.82(d, J=7.3Hz,2H),1.66(d,J=10.2Hz,4H),1.43(s,4H),1.15(s,26H),0.81-0.74(m,3H).

[0430] Preparation of 18 To a solution of 18-2 (5 g, 12.6 mmol), N-hydroxysuccinimide (2.9 g, 25.2 mmol, 2.0 equiv.), and 4-dimethylaminopyridine (310 mg, 2.5 mmol, 0.2 equiv.) in DMF (50 mL) was added N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.63 g, 18.9 mmol, 1.5 equiv.) at 0 °C under an inert atmosphere of nitrogen. The resulting solution was stirred at room temperature for 16 h and then diluted with 100 mL of dichloromethane. The organic layer was washed with 2 × 100 mL of water and 2 × 100 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (70 / 1 to 1 / 1). The product-containing fractions were concentrated and the residue was crystallized from dichloromethane / diethyl ether (30 / 1) to give 2.41 g (38% yield) of compound 18 as a white solid. MS: m / z 493.20 [M+H] + . 1 H NMR(CDCl3:)δ5.44(d,J=8.1Hz,1H),4.00-3.90(m,1H),2.98-2.91(m,1H),2.87(d,J=2.4Hz,4H ),2.20-2.07(m,4H),1.91-1.78(m,4H),1.68-1.52(m,4H),1.27(s,26H),0.90(t,J=6.8Hz,3H).

[0431] Example 19.

[0432] [ka]

[0433] Preparation of 19-1 To a solution of (-)-borneol (2 g, 12.7 mmol) in dichloromethane (30 mL) under N2, 16-(benzyloxy)-16-oxohexadecanoic acid (7.3 g, 19.44 mmol, 1.5 equiv.), 4-dimethylaminopyridine (0.48 g, 3.89 mmol, 0.3 equiv.), and dicyclohexylcarbodiimide (13.3 g, 64.83 mmol, 5.0 equiv.) were added and stirred overnight. The reaction mixture was then diluted with water and extracted with dichloromethane. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 0 to 50 / 1) to give 6.5 g (97% yield) of 19-1 as a yellow solid. MS: m / z 535.20 [M+Na] + .

[0434] Preparation of 19-2 To a solution of 19-1 (6.7 g, 13.06 mmol) in tetrahydrofuran (67 mL), 10% palladium on activated carbon (1.4 g) was added and flushed with five cycles of H2. The resulting mixture was stirred for 16 h, then filtered, the solid was washed with tetrahydrofuran, and the combined filtrate was concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (50 / 1 to 10 / 1) to give 4 g of 19-2 (75% yield) as a white solid. MS: m / z 445.40 [M+Na] + .

[0435] Preparation of 19 To a solution of 19-2 (5.5 g, 13.0 mmol) in N,N-dimethylformamide (55 mL), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (2.42 g, 15.6 mmol, 1.2 equiv.), N-hydroxysuccinimide (1.80 g, 16.6 mmol, 1.2 equiv.), and N,N-diisopropylethylamine (5.04 g, 39.1 mmol, 3 equiv.) were added and stirred for 16 h. The mixture was diluted with ethyl acetate. The organic layer was washed with water and brine and dried over anhydrous Na2SO4. The evaporated residue was purified on a silica gel column using hexane / ethyl acetate (100 / 0 to 15 / 1) to give 2.1 g (40% yield) of compound 19 as an off-white solid. MS: m / z 542.20 [M+Na] + . 1 H-NMR (DMSO-d6): δ4.81(t,J=5.2Hz,1H),2.81(s,4H),2.65(t,J=7.2Hz,2H),2.72-2.3 1(m,3H),1.88-1.84(m,1H),1.72-1.51(m,6H),1.34-1.14(m,23H),0.92-0.78(m,9H).

[0436] Example 20.

[0437] [ka]

[0438] Preparation of 20-2 To a solution of (1S,2R,4S)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-ol (16.8 g, 109.06 mmol, 2.0 equiv.) in N,N-dimethylformamide (250 mL) was added sodium hydride (2.6 g, 109.1 mmol, 2.0 equiv.) at 0 °C and stirred for 30 min. 20-1 (25 g, 54.5 mmol, 1.0 equiv.) was then added at 0 °C, and the solution was stirred at 40 °C for 16 h. The reaction was quenched with saturated NH4Cl solution and extracted with dichloromethane. The combined organic layer was washed with water and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 10 / 1) to give 4.5 g (17% yield) of 20-2 as a yellow oil.

[0439] Preparation of 20-3 To a solution of 20-2 (2.3 g, 4.74 mmol) in THF (46 mL), palladium on carbon (4.6 g) was added, and the mixture was stirred under H2 (20 atm pressure) at 65 °C for 8 h. The resulting mixture was filtered, and the filtrate was concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 6 / 1) to give 1.8 g (96% yield) of 20-3 as a white solid. 1 H NMR(CDCl3):δ3.63(t,J=6.6Hz,2H),3.53(ddd,J=9.5,3.4,1.9Hz,1H),3.46-3.27(m,2H),2.16-1.91( m,2H),1.63-1.47(m,7H),1.27(d,J=8.2Hz,25H),0.99(dd,J=13.0,3.4Hz,1H),0.85(d,J=7.5Hz,9H).

[0440] Preparation of 20-4 To a solution of 20-3 (3.46 g, 8.8 mmol) in acetone (121 mL), Jones reagent (10.5 mL) was added at 0 °C and stirred at 40 °C for 12 h. The reaction mixture was then filtered, concentrated, and purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 6 / 1) to give 2.87 g (80% yield) of 20-4 as a white solid. 1H NMR (DMSO-d6): δ12.29-11.36(m,1H),3.56-3.13(m,3H),2.15(t,J=7.4Hz,2H),2.10-1.90(m,2H),1.68-1.54(m,2H),1.4 7(h,J=6.6Hz,4H),1.24(d,J=11.0Hz,22H),1.11(ddt,J=17.3,11.8,4.9Hz,2H),0.97-0.88(m,1H),0.79(d,J=2.5Hz,9H).

[0441] Preparation of 20 To a solution of 20-4 (3.5 g, 8.6 mmol) in N,N-dimethylformamide (70 mL) at 0 °C, 1-hydroxypyrrolidine-2,5-dione (1.9 g, 17.1 mmol, 2.0 equiv.), 4-dimethylaminopyridine (0.21 g, 1.7 mmol, 0.2 equiv.), N,N-diisopropylethylamine (3.3 g, 25.7 mmol, 3.0 equiv.), and 3-(((ethylamine)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (2.4 g, 12.8 mmol, 1.5 equiv.) were added and stirred at room temperature for 16 h. The resulting mixture was extracted with dichloromethane. The organic layer was washed with water and brine, dried (anhydrous NaSO), filtered, and concentrated. The residue was purified on a silica gel column using n-hexane / ethyl acetate (100 / 1 to 6 / 1) to give 2.0 g (44% yield) of compound 20 as an off-white solid. MS: m / z 528.4 [M+Na] + . 1 H NMR(CDCl3):δ3.58-3.48(m,1H),3.46-3.26(m,2H),2.84(d,J=2.3Hz,4H),2.66-2.54(m,2H),2.16-1.91(m,2H),1.79-1.50(m ,7H),1.39(d,J=8.8Hz,2H),1.27(d,J=7.5Hz,20H),1.20-1.12(m,1H),1.02-0.95(m,1H),0.86(s,3H),0.83(d,J=1.1Hz,6H).

[0442] Example 21.

[0443] [ka]

[0444] Preparation of 22-1 A solution of 16-(benzyloxy)-16-oxohexadecanoic acid (4.8 g, 12.8 mmol) in sulfuryl dichloride (40 mL) was stirred at 70 °C for 1 h. It was then cooled to room temperature and concentrated. The crude product was added to a solution of (1R,2S,5R)-2-isopropyl-5-methylcyclohexan-1-ol (2 g, 12.79 mmol, 1.0 equiv.) in tetrahydrofuran (10 mL) at room temperature and stirred for 2 h. The pH was adjusted to 7 with 1 M NaOH solution and extracted with ethyl acetate. The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column (petroleum ether / ethyl acetate, 100 / 0 to 80 / 1) to give 5 g (78% yield) of 22-1 as a colorless oil. MS: m / z 537.50 [M+Na] + .

[0445] Preparation of 22-2 To a solution of 22-1 (5 g, 7.41 mmol) in tetrahydrofuran (50 mL) under N was added 10% palladium on activated carbon (1 g). The flask was flushed with H for five cycles and stirred under H for 16 h. The reaction mixture was then filtered and concentrated, and the residue was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 0 to 40 / 1) to give 3.5 g (74% yield) of compound 22-2 as a white solid. MS: m / z 447.40 [M+Na] + .

[0446] Preparation of 22 To a solution of 22-2 (5.5 g, 11.656 mmol) in N,N-dimethylformamide (55 mL), 1-hydroxypyrrolidine-2,5-dione (2 g, 17.48 mmol, 1.5 equiv.), 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (3.3 g, 17.48 mmol, 1.5 equiv.), and N,N-diisopropylethylamine (4.5 g, 34.96 mmol, 3 equiv.) were added and stirred for 16 h. The reaction mixture was extracted with ethyl acetate, and the organic layer was washed with water and brine. The organic layer was then dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (100 / 0 to 15 / 1) to give 2.7 g of compound 22 as a white solid. MS: m / z 544.40 [M+Na] + . 1 H NMR(DMSO-d6)δ4.61-4.54(m,1H),2.81(s,4H),2.65(t,J=7.2Hz,2H),2.25(t,J=7.2Hz,2H),1. 88-1.80(m,2H),1.66-1.49(m,6H)1.44-1.24(m,22H),1.11-0.83(m,9H),0.71(d,J=6.9Hz,3H).

[0447] Example 22.

[0448] [ka]

[0449] Preparation of 23-1 To a solution of 1,16-hexadecanediol (60 g, 232.56 mmol) in cyclohexane (600 mL) was added hydrogen bromide (43.2 mL, 255.8 mmol, 1.1 equiv). The mixture was warmed to reflux and stirred for 48 h. The resulting mixture was cooled to room temperature, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (60 / 1 to 30 / 1) to give 35 g (47% yield) of 23-1 as a white solid.

[0450] Preparation of 23-2 To a solution of 23-1 (35 g, 109.4 mmol) in anhydrous N,N-dimethylformamide (700 mL) was added sodium hydride (60% dispersion in oil, 8.75 g, 218.8 mmol, 2.0 equiv.) at 0 °C under Ar and stirred for 30 min. Benzyl bromide (28.06 g, 164.1 mmol, 1.5 equiv.) was added to the reaction mixture at 0 °C and stirred at room temperature for 16 h. The reaction mixture was quenched with saturated NH4Cl, diluted with ethyl acetate, and washed with water and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (500 / 1 to 200 / 1) to give 30 g (67% yield) of 23-2 as a white solid. 1 H NMR (DMSO-d6): δ7.42-7.23(m,5H),4.48(d,J=39.7Hz,2H),3.58-3.36(m,4H),1.78(p,J=6.8Hz,2H),1.52(p,J=6.7Hz,2H),1.23(s,24H).

[0451] Preparation of 20-1 To a mixture of 23-2 (30 g, 93.75 mmol, 1.0 equiv) in acetone (600 mL), sodium iodide (28.125 g, 187.5 mmol, 2.0 equiv) was added and refluxed with vigorous stirring for 2 h. The reaction mixture was cooled, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 60 / 1) to give 30 g (89.5% yield) of compound 20-1 as a white solid. 1 H NMR(CDCl3):δ7.35-7.21(m,5H),4.50(d,J=2.1Hz,2H),3.46(t,J=6.6Hz,2H),3 .18(t,J=7.0Hz,2H),1.82(p,J=7.1Hz,2H),1.61(p,J=6.8Hz,2H),1.26(s,24H).

[0452] Preparation of 23-3 To a solution of L(-)-menthol (20.44 g, 131.0 mmol, 2.0 equiv) in anhydrous N,N-dimethylformamide (409 mL) under Ar at 0 °C, sodium hydride (60% dispersion in oil, 5.24 g, 131.0 mmol, 2.0 equiv) was added and stirred for 30 min. 20-1 (30 g, 65.50 mmol, 1.0 equiv) was then added to the reaction mixture at 0 °C and stirred at room temperature for 16 h. The reaction mixture was quenched with saturated NH4Cl, diluted with ethyl acetate, and washed with water and brine. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (200 / 1 to 100 / 1) to give 9.8 g (30% yield) of 23-3 as a colorless oil. 1 H NMR(CDCl3):δ7.39-7.22(m,5H),4.50(s,2H),3.60(dt,J=9.1,6.4Hz,1H),3.46(t,J=6.6Hz,2H),3.38(t,J=6.7Hz,1H),3.25(dt,J=9.2,6.9Hz,1H ),2.98(td,J=10.5,4.1Hz,1H),2.23(dqd,J=14.1,7.2,2.7Hz,1H),2.14 -2.00(m,1H),1.69-1.51(m,6H),1.35-1.23(m,27H),0.94-0.75(m,10H).

[0453] Preparation of 23-4 To a solution of 23-3 (7.2 g, 14.81 mmol, 1.0 equiv) in ethyl acetate (144 mL) under H2, palladium on activated carbon (10%, 14.4 g) and acetic acid (0.89 g, 14.8 mmol, 1.0 equiv) were added and stirred at 65 °C for 8 h. The solution was filtered and concentrated, and the residue was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 50 / 1) to give 5.2 g (88.6% yield) of 23-4 as a white solid. 1H NMR(CDCl3)δ3.62(dt,J=15.5,6.5Hz,3H),3.25(dt,J=9.2,6.9Hz,1H),2.99(td,J=10.6,4.1Hz,1H),2. 21(pt,J=9.6,4.8Hz,1H),2.14-2.04(m,1H),1.70-1.47(m,6H),1.38-1.14(m,27H),1.04-0.74(m,12H).

[0454] Preparation of 23-5 To a solution of 23-4 (5.2 g, 13.13 mmol, 1.0 equiv) in acetone (182 mL) under Ar at 0 °C, Jones reagent (2.67 M, 15.74 mL, 42.02 mmol, 3.2 equiv) was added and stirred at 40 °C for 12 h. The reaction mixture was filtered and concentrated, and the residue was diluted with dichloromethane and then washed with water and brine. The organic layer was dried over anhydrous NaSO, filtered, and concentrated. The residue was purified on a silica gel column using petroleum ether / ethyl acetate (40 / 1 to 8 / 1) to give 3.5 g (65% yield) of 23-5 as an aqueous solid. MS: m / z 409.30 [M−H] - . 1 H NMR(CDCl3)δ3.62-2.82(m,3H),2.27(t,J=7.5Hz,2H),2.21-1.96(m,2H),1.65-1.39(m,6H),1.21(d,J=13.8Hz,25H),0.94-0.66(m,11H).

[0455] Preparation of 23 To a solution of 23-5 (3.5 g, 8.54 mmol, 1.0 equiv.) and N-hydroxysuccinimide (1.96 g, 17.07 mmol, 2.0 equiv.) in N,N-dimethylformamide (70 mL) was added 4-dimethylaminopyridine (0.21 g, 1.71 mmol, 0.2 equiv.) and N,N-diisopropylethylamine (3.3 g, 25.61 mmol, 3.0 equiv.) under Ar at 0 °C and stirred for 30 min. Then, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (2.46 g, 12.81 mmol, 1.5 equiv.) was added and stirred at room temperature for 12 h. The reaction mixture was diluted with ethyl acetate and washed with water and brine. The organic layer was dried over anhydrous NaSO, filtered, and concentrated. The residue was purified on a silica gel column using hexane / ethyl acetate (50 / 1 to 6 / 1) to give 2.0 g (46.9% yield) of compound 23 as a white solid. MS: m / z 508.45 [M+H] + . 1 H NMR(CDCl3)δ3.60(dt,J=9.1,6.3Hz,1H),3.24(dt,J=9.1,6.8Hz,1H),2.98(td,J=10.5,4.1Hz,1H),2.83(d,J=2.5Hz,4H),2.60(t,J=7.5Hz,2H),2. 22(pd,J=6.9,2.7Hz,1H),2.14-2.02(m,1H),1.74(p,J=7.4Hz,2H),1.67- 1.47(m,4H),1.26(d,J=5.7Hz,24H),1.04-0.83(m,8H),0.82-0.73(m,4H).

[0456] Example 23.

[0457] [ka]

[0458] Preparation of 24-1 To a solution of 1,1'-carbonyldiimidazole (4.8 g, 19.6 mmol, 1.8 equiv.) in 50 mL of dichloromethane was added hexadecane-1-thiol (6.38 g, 24.69 mmol, 1.5 equiv.) at room temperature under an inert atmosphere of nitrogen. The resulting solution was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure. The crude product was dissolved in dichloromethane (10 mL) and then added dropwise to acetonitrile (100 mL). The solid was collected by filtration. The cake was dissolved in 20 mL of N,N-dimethylformamide and 20 mL of tetrahydrofuran. 2'-amino-D-uridine (4 g, 16.5 mmol, 1.0 equiv.) and 1-hydroxybenzotriazole (3.33 g, 24.7 mmol, 1.5 equiv.) were then added at room temperature. The resulting solution was stirred at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure. The crude product was purified by preparative flash chromatography using the following conditions: C18 column, mobile phase: water and tetrahydrofuran (30% to 100% THF gradient). The product-containing fractions were concentrated to give 6.9 g (80% yield) of 24-1 as a white solid. MS: m / z 528.15 [M+H] + . 1 H NMR(DMSO-d6)δ11.30(d,J=2.3Hz,1H),8.05(d,J=8.4Hz,1H),7.87(d,J=8.1Hz ,1H),5.90(d,J=8.2Hz,1H),5.75-5.59(m,2H),5.16(t,J=5.0Hz,1H),4.55-4.4 0(m,1H),4.08(t,J=5.7Hz,1H),3.90(q,J=2.8Hz,1H),3.66-3.49(m,2H),2.84 -2.63(m,2H),1.45(d,J=7.0Hz,2H),1.23(d,J=2.8Hz,26H),0.92-0.79(m,3H).

[0459] Preparation of 24-2 To a solution of 24-1 (4 g, 7.59 mmol) in 40 mL of pyridine was added 4,4'-dimethoxytrityl chloride (3.1 g, 9.1 mmol, 1.2 equiv.) at 0 °C under an inert atmosphere of argon. The resulting solution was stirred at 25 °C for 3 h. The residue was diluted with 300 mL of dichloromethane and quenched with saturated aqueous sodium bicarbonate solution. The mixture was washed with 2 × 150 mL of saturated aqueous sodium bicarbonate solution and 2 × 150 mL of saturated brine. The organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by preparative flash chromatography using the following conditions: C18 column, mobile phase, water and THF, gradient 30% to 100% THF. The product-containing fractions were combined and diluted with an equal volume of dichloromethane. The organic layer was separated. The aqueous layer was extracted with 3 × 200 mL of dichloromethane. The combined organic extracts were dried (anhydrous NaSO), filtered, and concentrated to give 3.9 g (62% yield) of 24-2 as a white solid. MS: m / z 852.45 [M+Na] + . 1 H NMR (DMSO-d6): δ11.36(s,1H),8.18(d,J=8.3Hz,1H),7.65(d,J=8.1Hz,1H),7.45-7.17( m,9H),6.96-6.84(m,4H),5.89(d,J=7.6Hz,1H),5.68(d,J=4.9Hz,1H),5.41(d,J=8.1Hz ,1H),4.64(d,J=6.4Hz,1H),4.19(d,J=7.1Hz,1H),4.00(d,J=3.5Hz,1H),3.75(s,6H),3 .31-3.15(m,2H),2.84-2.70(m,2H),1.58-1.40(m,2H),1.24(s,26H),0.91-0.75(m,3H).

[0460] Preparation of 24 To a solution of 24-2 (3.1 g, 3.74 mmol) in dichloromethane (31 mL) was added 3-(bis(diisopropylamino)phosphinooxy)propanenitrile (1.35 g, 4.48 mmol, 1.2 equiv.) and 4,5-dicyanoimidazole (485 mg, 4.1 mmol, 1.1 equiv.) under an inert atmosphere of argon, and the reaction mixture was stirred at room temperature for 2 h. The resulting solution was diluted with 500 mL of dichloromethane and washed with 300 mL of saturated aqueous sodium bicarbonate solution and 300 mL of saturated brine, respectively. The organic layer was dried (anhydrous Na2SO4), filtered, and concentrated. The crude product was purified by preparative flash chromatography using the following conditions: C18 column, mobile phase, water and THF, gradient 30% to 100% THF. The product-containing fractions were combined and diluted with an equal volume of dichloromethane. The organic layer was separated. The aqueous phase was extracted with 3 x 200 mL of dichloromethane. The combined organic extracts were dried (anhydrous NaSO), filtered, and concentrated. The crude residue was purified on a silica gel column using hexane / ethyl acetate (5 / 1 to 2 / 1) to give 2.624 g (69% yield) of compound 24 as a white solid. MS m / z 1030.55 = [M+H] + . 1 H NMR(DMSO-d6)δ11.43(s,1H),8.44(dd,J=33.8,8.3Hz,1H),7.69(dd,J=8.2,5.0Hz,1H),7.40(dd,J=7.7,2.5Hz,2H),7.35-7.21 (m,7H),6.89(dd,J=9.0,3.0Hz,4H),5.94(t,J=7.0Hz,1H),5.45(d,J=7.9Hz,1H),4.90-4.70(m,1H),4.47-4.28(m,1H),4.26-4 .11(m,1H),3.93-3.77(m,1H),3.74(s,6H),3.72-3.61(m,1H),3.60-3.43(m,2H),3.30-3.18(m,2H),2.92-2.79(m,1H),2.78-2 .68(m,2H),2.63(t,J=6.0Hz,1H),1.58-1.46(m,2H),1.23(s,26H),1.15-1.05(m,9H),0.96(d,J=6.7Hz,3H),0.88-0.82(m,3H). 31P NMR(DMSO-d6)δ149.13,147.57.

[0461] Example 24.

[0462] [ka]

[0463] Preparation of 25-1 To a solution of 17-6 (14 g, 38.60 mmol), triethylamine (11.7 g, 115.8 mmol, 3.0 equiv.), and dimethylaminopyridine (2.4 g, 19.30 mmol, 0.5 equiv.) in 140 mL of dichloromethane was added 4-methylbenzenesulfonyl chloride (8.1 g, 42.5 mmol, 1.1 equiv.) at °C. The resulting solution was stirred at room temperature for 2 h. The reaction was quenched by the addition of 500 mL of saturated sodium bicarbonate solution, and the resulting mixture was extracted with 3 × 800 mL of dichloromethane. The combined organic layers were washed with 2 × 500 mL of water and 500 mL of brine, dried over anhydrous NaSO, filtered, and concentrated. A mixture of the crude product and sodium iodide (28.9 g, 193.0 mmol, 5.0 equiv.) in 150 mL of acetone was refluxed with vigorous stirring for 2 h, then cooled to room temperature and extracted with 3 × 800 mL of dichloromethane. The combined organic layers were washed with 2 × 500 mL of water and 500 mL of saturated brine, dried over anhydrous NaSO, filtered, and concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 10 / 1) to give 14.2 g (78% yield) of 25-1 as a colorless oil. 1 H-NMR (CDCl3): δ3.09(t,J=7.0Hz,2H),1.83(t,J=3.2Hz,3H),1.71(q,J=7.1Hz,2H),1.63-1.40(m,6H),1.38-1.05(m,30H),0.93(d,J=7.2Hz,2H).

[0464] Preparation of 25-3 To a solution of 25-2 (3.1 g, 7.4 mmol) in 70 mL of tetrahydrofuran was added sodium hydride (0.71 g, 29.6 mmol, 4.0 equiv.) at 0 °C, and the resulting mixture was stirred at 0 °C for 30 min. 25-1 (13.9 g, 29.6 mmol, 4.0 equiv.) was then added at 0 °C, and the reaction mixture was stirred at 65 °C for 6 h. The reaction was quenched by the addition of saturated ammonium chloride solution (500 mL) and extracted with dichloromethane (3 × 800 mL). The combined organic layers were washed with water and brine, dried over anhydrous NaSO, filtered, and concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 1 / 1) to give 2.40 g (42% yield) of 25-3 as a white solid. MS m / z 791.50 = [M + Na] + .

[0465] Preparation of 25-4 To a solution of 25-3 (3.7 g, 4.811 mmol, 1.0 equiv) in methanol (37 mL) and tetrahydrofuran (37 mL), palladium hydroxide (3.7 g) was added and stirred under a H atmosphere for 2 h. The mixture was filtered, and the filtrate was concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 3 / 1) to give 2.1 g (74% yield) of 25-4 as a white solid. 1 H-NMR (DMSO-d6): δ11.32(s,1H),7.94(d,J=8.1Hz,1H),5.84(d,J=5.1Hz,1H),5.63(d,J=8.1Hz, 1H),5.12(t,J=5.0Hz,1H),5.02(d,J=5.8Hz,1H),4.12-4.02(m,1H),3.90-3.80(m,2H),3.69-3. 51(m,3H),3.44(dddd,J=11.9,6.9,5.0,1.7Hz,1H),3.17(d,J=5.2Hz,1H),1.90(p,J=2.9,2.3Hz ,3H),1.70-1.53(m,6H),1.53-1.38(m,8H),1.22(d,J=1.8Hz,23H),1.00(dt,J=12.8,4.2Hz,2H).

[0466] Preparation of 25-5 To a solution of 25-4 (1.0 g, 1.698 mmol) in pyridine (10 mL) was added dimethoxytrityl chloride (748 mg, 2.21 mmol, 1.3 equiv.) at 0 °C, followed by stirring at room temperature for 12 h. The resulting solution was extracted with 3 × 100 mL of dichloromethane. The organic layer was washed with 2 × 100 mL of water and 100 mL of saturated brine, dried over anhydrous NaSO, filtered, and concentrated. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (100 / 1 to 1 / 2) to give 1.3 g (85% yield) of 25-5 as a white solid. 1 H-NMR (DMSO-d6): δ11.38(d,J=1.9Hz,1H),7.73(d,J=8.1Hz,1H),7.47-7.10(m,9H),6.95-6.78( m,4H),5.80(d,J=3.7Hz,1H),5.28(dd,J=8.1,1.8Hz,1H),5.11(d,J=6.5Hz,1H),4.22-3.84(m,4 H),3.74(s,6H),3.57(qt,J=9.2,6.3Hz,2H),3.26(td,J=12.3,10.7,3.5Hz,2H),1.88(q,J=3.1H z,3H),1.70-1.45(m,8H),1.41(d,J=2.8Hz,6H),1.20(d,J=12.8Hz,23H),1.00(d,J=7.2Hz,2H).

[0467] Preparation of 25 To a solution of 25-5 (3.3 g, 3.7 mmol) in dichloromethane (33 mL) were added 2-cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite (1.9 g, 6.3 mmol, 1.7 equiv.) and 4,5-dicyanoimidazole (0.5 g, 4.1 mmol, 1.1 equiv.). The resulting solution was stirred at room temperature for 2 h and then diluted with 3 × 500 mL of dichloromethane. The combined organic extracts were washed with 2 × 500 mL of saturated brine, dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified on a silica gel column using n-hexane / ethyl acetate (100 / 1 to 1 / 1) to give 2.38 g (57% yield) of compound 25 as a white solid. MS: m / z 1091.55 [M+H] + . 1H-NMR(DMSO-d6): δ11.39(s,1H),7.86-7.73(m,1H),7.45-7.17(m,9H),6.95 -6.81(m,4H),5.80(t,J=3.1Hz,1H),5.24(t,J=8.9Hz,1H),4.47-4.26(m,1H) ,4.18-3.97(m,2H),3.83-3.43(m,12H),3.23-3.15(m,2H),2.82-2.56(m,2H) ,1.88(s,3H),1.71-1.35(m,14H),1.29-1.04(m,33H),0.98(d,J=6.8Hz,5H). 31 PNMR (DMSO-d6): δ149.13,148.53.

[0468] Example 25. The following compounds were prepared in a similar manner to the compounds provided in the previous examples using the corresponding starting materials.

[0469] [Table 3-1]

[0470] [Table 3-2]

[0471] [Table 3-3]

[0472] Example 26. Knockdown efficiency of MAPT siRNA in human iPSC-neurons. MAPT siRNA agents conjugated with specific lipophilic monomers provided herein were synthesized and the sequences are listed in Table 3.

[0473] Table 3: Modified sense and antisense strand sequences of MAPT siRNA conjugated with different lipids (5' to 3') Modifications: mN = 2'OMe; fN = 2'F; ps = phosphorothioate; VP = vinylphosphonate; invAb = reverse abasic Chol4 = cholesterol-triethylene glycol

[0474] [Table 4-1]

[0475] [Table 4-2]

[0476] [Table 4-3]

[0477] [Table 4-4]

[0478] [Table 4-5]

[0479] The in vitro knockdown efficiency of the lipid-siRNA conjugates was evaluated in human iPSC-derived cortical neurons according to the procedure described below.

[0480] Differentiation of human iPSCs into cortical neurons A human iPSC line (Sigma#iPSC0028) was derived by OSKM retroviral reprogramming of epithelial cells from a 24-year-old Caucasian female donor. iPSCs were first differentiated into cortical neural stem cells (NSCs) according to a dual SMAD inhibitor protocol (Shi et al., 2012, 2012, Nat. Proc. 7(10):1836-46) with some modifications. Briefly, iPSCs were plated at 500,000 cells / cm on Matrigel (Corning 354230)-coated wells in mTeSR medium (StemCell Technologies 5850) supplemented with 10 μM ROCK inhibitor (Sigma-Aldrich Y0503). 2and cultured at 37°C and 5% O. The following day (day -1), the medium was replaced with mTeSR. From day 0 to day 12, cell medium was replaced daily with cortical neural induction medium containing 10 μM SB43142 (Tocris 1614) and 1 μM dorsomorphin (Tocris 3093), and cultured in Neural Maintenance Medium (1:1 DMEM:F12 supplemented with Glutamax (ThermoFisher Scientific 10565108), Neurobasal (ThermoFisher Scientific 21103049), 2.5 μg / mL insulin (Sigma-Aldrich I9278-5ML), 50 μM 2-mercaptoethanol (ThermoFisher Scientific, 31350010), 0.5% non-essential amino acids (ThermoFisher Scientific 11140035), 0.5% GlutaMAX supplement (ThermoFisher Scientific 11140035), and 0.5% GlutaMAX supplement (ThermoFisher Scientific 11140035). The media was supplemented with 0.5 mM sodium pyruvate (ThermoFisher Scientific 10565018), 0.5 mM sodium pyruvate (ThermoFisher Scientific 11360070), 1% penicillin-streptomycin (Sigma P4333), 0.5% N2 supplement (ThermoFisher Scientific 17502048), and 1% B27 supplement (ThermoFisher Scientific 17504044). On day 12, neuroepithelial sheets were gently dissociated into large aggregates of 300-500 cells using a needle and lifter, and the aggregates were collected in a 15 mL falcon by centrifugation at 160 g for 2 min. The media was supplemented with 10 μg / mL laminin (Sigma-Aldrich The cell pellet was gently resuspended in neural induction medium in a total volume of 2 mL per well of a 6-well plate for passage 1 / 2 or 1 / 3 onto laminin-coated wells. The medium was replaced with neural maintenance medium supplemented with 20 ng / mL FGF2 (Stemcell Technologies 2634) on days 13 and 15. On day 17, neural rosettes were detached at passage 1 / 3 using dispase (ThermoFisher Scientific 17105041) and plated onto laminin-coated wells.One or two additional dispase steps were performed for an additional week. Approximately on days 25-30, neural stem cells were dissociated into single-cell suspensions with Accutase (ThermoFisher Scientific A1110501) and cryopreserved in freshly prepared neural freezing medium containing neural maintenance medium supplemented with 10% (v / v) DMSO and 20 ng / mL FGF2. Frozen vials of neural stem cells were stored in liquid nitrogen until use.

[0481] To generate iPSC-neurons, thaw a frozen vial of neural stem cells (NSCs) and plate them at 70,000 cells / cm on laminin-coated wells in neural maintenance medium supplemented with 10 μM ROCK inhibitor and 20 ng / mL FGF2. 2 The cells were seeded at 100°C for 2 days. The medium was replaced daily with neural maintenance medium for the next 2 days. Approximately 4 days after thawing, cells were dissociated with Accutase and seeded at 28,000 cells per well onto 96-well plates pre-coated with poly-L-ornithine and laminin in neural maintenance medium supplemented with 10 μM ROCK inhibitor. The day after reseeding, the culture medium was replaced with neural differentiation medium consisting of neural maintenance medium supplemented with 20 ng / mL BDNF (R&D Systems 212-BD-050 / CF), 20 ng / mL GDNF (R&D Systems 212-GD-050 / CF), 500 μM DB-cAMP (Sigma D0627), and 20 mM ascorbic acid (Sigma A4403). Cultures were differentiated in neural differentiation medium with 50% medium changes twice weekly. Two to three weeks after differentiation from neural stem cells (NSCs), neurons were treated with siRNA for 7 or 14 days for RNA analysis by reverse transcription and real-time PCR and protein analysis by MSD immunoassay.

[0482] MSD Immunoassay Human iPSC neurons differentiated on 96-well plates were lysed in 100 μL of ice-cold RIPA buffer (Sigma) supplemented with cOmplete™ Protease Inhibitor Cocktail (Roche) and PhosSTOP™ (Roche) for 30 minutes at 4°C with slow orbital shaking. Plates were centrifuged at 1,000 × g for 5 minutes, and cell lysates were collected and diluted at different times (20, 50, or 100) for protein measurement using MSD immunoassays according to standardized procedures. Briefly, MSD plates were coated overnight at 4°C with 30 μL / well of coating antibody diluted in PBS at 1 μg / mL. The next day, plates were inverted onto absorbent tissue to dry and then incubated with 150 μL per well of blocking buffer 0.1% casein for 2 hours at room temperature with orbital shaking at 300 rpm. After blocking, the plates were incubated overnight with cell lysates diluted in RIPA buffer supplemented with cOmplete™ Protease Inhibitor Cocktail (Roche) at 4°C with slow orbital shaking. The plates were washed five times using a Titertek Aquamax 4000 and dried upside down on absorbent tissue. Then, 25 μL of secondary or detection antibody diluted in 0.1% casein was added to the MSD plate and incubated for 2 hours at room temperature. After antibody incubation, the plates were washed five times and developed with 150 μL / well of 2x MSD read buffer (Meso Scale Discovery) diluted in milliQ H2O. The plates were immediately read using the MSD instrument. For tau protein measurement, hTau43 antibody (Janssen) was used to coat the 96-well MSD plate, and SULFO-TAG™-labeled hTau60 antibody (Janssen) was used for detection. For the measurement of histone H3 protein, recombinant rabbit monoclonal antibody 17H2L9 (ThermoFisher) was used for plate coating, mouse mAb 14221 BF (Cell Signaling Technology) was used as primary detection, and SULFP-TAG-labeled anti-mouse antibody (Meso Scale Discovery) was used as secondary detection.

[0483] Tables 4-6 summarize the efficiency of MAPT siRNA conjugated with different lipids in knocking down MAPT mRNA in human iPSC neurons after 7 days of incubation. Three concentrations of siRNA conjugates were tested: 1 μM, 200 nM, and 40 nM. Data are shown as residual MAPT mRNA relative to the control. Mean ± SD, n=3.

[0484] [Table 5]

[0485] [Table 6]

[0486] [Table 7]

[0487] Example 27. Stability study The in vitro stability of the siRNA conjugates was evaluated in various matrices, including mouse brain homogenate, human liver lysosomes, and rat liver tritosomes, using the methods described below. The analytical results are shown in Tables 7 and 8.

[0488] Tables 7 and 8 summarize LC-MS measurements of the antisense and sense strand stability of siRNA conjugates in mouse brain homogenates (Tables 7 and 10), human liver lysosomes (Tables 8 and 11), and rat liver tritosomes (Table 12) at 37° C. after 24 hours of incubation with shaking at 450 rpm. To assess the stringency of the assay conditions, the reference compound MSC-2-V was added to the assay.

[0489] [Table 8]

[0490] [Table 9]

[0491] [Table 10]

[0492] [Table 11]

[0493] [Table 12]

[0494] [Table 13]

[0495] [Table 14]

[0496] Example 28: Evaluation of in vivo knockdown efficiency of siRNA conjugates in a mouse model The in vivo knockdown efficiency of lipid-siRNA conjugates was evaluated in a mouse model using intracerebroventricular (ICV) injection according to the procedure described below.

[0497] MAPT mRNA was assessed in seven brain regions (cortex, hippocampus, brainstem, cerebellum, striatum, midbrain, and cervical spinal cord) from hTAU KI mice 7 days after a single ICV injection of M28_Var1, M28_Var39, M28_Var40, M28_Var41, M28_Var42, M28_Var50 at 15 nmol. Results are shown in Table 9. Data are presented as residual MAPT mRNA (%). Mean ± SD, n=6 mice per treatment group.

[0498] Mice and intracerebroventricular injection Male and female PS19 mice (C57BL6; Prnp-MAPT * P301S or hTauKI (C57BL6; hMAPT: knockin) mice (2-3 months old) were randomly assigned to different treatment groups. Mice were anesthetized with isoflurane (induction: 4-5%; maintenance: 1.8-2.5%). They were then stereotactically injected into both ventricles at coordinates: AP: -0.62 mm, ML: + / - 1.05 mm, DV: 2.2 mm, using an electric drill and a microinjection robot (Neurostar, Germany, Sterodrive Software v 2019). Each injection was performed with a volume of 5 µL over 5 min. After injection, the needle was withdrawn in three steps (1.1 mm, held for 60 s, held for 5 min; 2. 0.5 mm further in 30 s, wait for 5 min; 3. Withdrawal from the brain at a very slow speed) to avoid backflow of the compound along the needle tract. After each step, the amount of backflow was checked. On selected days after injection, animals are sacrificed and different brain regions, including the cortex, hippocampus, brainstem, cerebellum, striatum, midbrain, and cervical spinal cord, are dissected, snap frozen, and stored at -80°C until further analysis.

[0499] RNA extraction from tissues The tissue was collected in a 2 mL tube containing 1.4 mm ceramic spheres (Lysing Matrix D (MP Biomedicals 6913-500)) and stored in a -80°C freezer until analysis. Place the tissue on ice under laminar flow and immediately add 750 μL of Trizol (ThermoFisher 15596026 / 15596018) per tube. Disrupt and homogenize the tissue using a FastPrep-24™ 5G Grinder at a speed of 5 m / s for three 30-second cycles. Chill the sample on ice for 2 minutes between cycles. After homogenization, briefly spin the tube and add 20% volume of chloroform (150 μL if 750 μL of Trizol was used). Vortex for 15 seconds and centrifuge at 14,000 × g for 15 minutes at 4°C. The upper aqueous phase is transferred to a deep 96-well plate, one volume of 70% ethanol is added, and the mixture is mixed thoroughly. The next step is performed using the RNeasy96 kit (Qiagen) according to the manufacturer's protocol. The RNeasy96 plate is placed on a square-well block holder, and the sample (Trizol / chloroform extraction) is applied to the wells of the RNeasy96 plate. The plate is sealed with an AirPore cover to prevent contamination. Centrifuge at 5600 x g for 3 minutes at room temperature, and discard the solution. Successive wash steps, including one 800 µL RW1 buffer, two 800 µL RPE buffer applied to the RNeasy96 plate, and wash buffer, are performed by centrifugation at 5600 x g for 3 minutes each. After the final wash, the RNeasy96 plate is centrifuged at 5600 x g for 3 minutes to remove residual lipids. RNA is eluted using 60 µL of RNase-free water by centrifugation at 5600 x g for 3 minutes at room temperature. RNA concentration is measured by Nanodrop8000 (ThermoFisher). Eluted RNA is stored at -80°C until analysis.

[0500] RT-qPCR RNA was used as a template for reverse transcription using High-Capacity cDNA Reverse Transcription Kits (Applied Biosystems) according to the manufacturer's protocol. Briefly, the final reaction mixture of 20 μL was incubated at 25°C for 10 minutes, followed by reverse transcription at 37°C for 2 hours and enzyme inactivation at 85°C for 5 minutes. For the qPCR reaction, the reverse-transcribed cDNA was diluted 10-fold and mixed with 2X PowerUp™ SYBR™ Green Mater Mix (ThermoFisher A25743) and 500 nM qPCR primers to a final reaction volume of 10 μL. qPCR was performed using a QuantStudio™ 12K instrument (Applied Biosystems™) following standard thermal cycling protocols. MAPT mRNA was detected using multiple primers, and reference primers targeting the mouse housekeeping genes AP3D1 and PAK1IP1 were included for gene expression normalization.

[0501] The embodiments described above are intended to be illustrative only; those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific compounds, materials, and procedures. All such equivalents are considered to be within the scope of this invention and included in the following claims.

Claims

1. A compound of formula (I), 【Chemistry 1】 Here, X is either O or S, Y is O or NR ａ And, D １ is H, a hydroxyl protecting group, or a reactive phosphorus group. D ２ is H, a hydroxyl protecting group, or a reactive phosphorus group. Or, D １ and D ２ Together, they form protecting groups for both the oxygen and Y to which they are bonded. Z １ 、Z ２ 、and Z ３ are each independently -O-, -NR ａ -, ＊ -OC(=O)-, ＊ -NR ａ C(=O)-, -OC(=O)O-, ＊ -NR ａ C(=O)O-, ＊ -OC(=O)NR ｂ - ＊ -NR ａ C(=O)NR ｂ - ＊ -OC(=S)-, ＊ -NR ａ C(=S)-, -OC(=S)O-, ＊ -NR ａ C(=S)O-, ＊ -OC(=S)NR ｂ - ＊ -NR ａ C(=S)NR ｂ - ＊ -OC(=O)S-, ＊ -NR ａ C(=O)S-, ＊ -OS(O) ｘ or ＊ -NR ａ S(O) ｘ -, and ＊ refers to the direction towards the ring containing X R １ , R ２ , and R ３ These are H and C, respectively, independently. １ ~C ６ C is optionally substituted with an alkoxy. １ ~C ６ Alkyl, or -(G-L) ｍ -R, however, R １ , R ２ , and R ３ At least one of them is -(G-L) ｍ -R is, Each example in G is independent of C １ ~C ８ It is alkylene, Each example of L is, independently, -O-, -NR ａ -, -C(=O)-, ＊ -OC(=O)-, ＊ -C(=O)O-, ＊ -NR ａ C(=O)-, ＊ -C(=O)NR ｂ -, -OC(=O)O-, ＊ -NR ａ C(=O)O-, ＊ -OC(=O)NR ｂ -, ＊ -NR ａ C(=O)NR ｂ -, ＊ -OC(=S)-, ＊ -C(=S)O-, ＊ -NR ａ C(=S)-, ＊ -C(=S)NR ｂ -, -OC(=S)O-, ＊ -NR ａ C(=S)O-, ＊ -OC(=S)NR ｂ -, ＊ -NR ａ C(=S)NR ｂ -, ＊ -OC(=O)S-, ＊ -SC(=O)O-, ＊ -NR ａ C(=O)S-, ＊ -SC(=O)NR ｂ -, ＊ -OS(O) ｘ -, ＊ -S(O) ｘ O-, ＊ -NR ａ S(O) ｘ - or ＊ -S(O) ｘ NR ｂ - and ＊ refers to the direction towards the ring containing X. R ａ Each example of １ to C ６ is alkyl and R ｂ Each of these examples is independently H or C １ ~C ６ It is alkyl, Each example in R is independent of C １２ ~C ３２ Alkyl or C １２ ~C ３２ It is alkenyl, x is either 1 or 2, n is either 0 or 1, A compound in which m is 0, 1, or 2. or its stereoisomer, or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein X is O or S.

3. The compound according to claim 1, which is a compound of formula (I-A), 【Chemistry 2】 or its stereoisomer, or a pharmaceutically acceptable salt thereof.

4. The compound according to claim 1, which is a compound of formula (I-B), 【Transformation 6】 or its stereoisomer, or a pharmaceutically acceptable salt thereof.

5. A compound of formula (III), 【Chemistry 10】 Here, B is a modified or unmodified nucleic acid base, X is either O or S, Y is O or NR ａ And, D １ is H, a hydroxyl protecting group, or a reactive phosphorus group. D ２ is H, a hydroxyl protecting group, or a reactive phosphorus group. Or, D １ and D ２ Together, they form protecting groups for both the oxygen and Y to which they are bonded. Z ２ is -O-, -NR ａ - ＊ -OC (=O)- ＊ -NR ａ C(=O)-, -OC(=O)O-, ＊ -NR ａ C(=O)O-, ＊ -OC(=O)NR ｂ - ＊ -NR ａ C(=O)NR ｂ - ＊ -OC(=S)- ＊ -NR ａ C(=S)-, -OC(=S)O-, ＊ -NR ａ C(=S)O-, ＊ -OC(=S)NR ｂ - ＊ -NR ａ C(=S)NR ｂ - ＊ -OC(=O)S-, ＊ -NR ａ C(=O)S- ＊ -OS(O) ｘ -, or ＊ -NR ａ S(O) ｘ - and ＊ This indicates the direction toward the ring containing X, R ２ is -(G-L) ｍ -R is, Each example in G is independent of C １ ~C ８ Alkylene or C ３ ~C １０ It is a cycloalkylene, Each example of L is independently -O-, -NR ａ -, -C(=O)-, ＊ -OC (=O)- ＊ -C(=O)O- ＊ -NR ａ C(=O)- ＊ -C(=O)NR ｂ -, -OC(=O)O-, ＊ -NR ａ C(=O)O-, ＊ -OC(=O)NR ｂ - ＊ -NR ａ C(=O)NR ｂ - ＊ -OC(=S)- ＊ -C(=S)O- ＊ -NR ａ C(=S)- ＊ -C(=S)NR ｂ -, -OC(=S)O-, ＊ -NR ａ C(=S)O-, ＊ -OC(=S)NR ｂ - ＊ -NR ａ C(=S)NR ｂ - ＊ -OC(=O)S-, ＊ -SC(=O)O-, ＊ -NR ａ C(=O)S- ＊ -SC(=O)NR ｂ - ＊ -OS(O) ｘ - ＊ -S(O) ｘ O-, ＊ -NR ａ S(O) ｘ -, or ＊ -S(O) ｘ NR ｂ - and ＊ This refers to the direction toward the ring containing X, R ａ Each of these examples is independently H or C １ ~C ６ It is alkyl, R ｂ Each of these examples is independently H or C １ ~C ６ It is alkyl, R is C １２ ~C ３２ Alkyl or C １２ ~C ３２ It is alkenyl, x is either 1 or 2, m is 0, 1, or 2, However, X is O, Y is O, Z ２ If it is -O-, (i) R is -L"-(CH ２ ) ０－３ It is replaced with -R, and here, L represents non-existence, -O-, -NR ａ -, -C(=O)-, ＊ -OC (=O)- ＊ -C(=O)O- ＊ -NR ａ C(=O)- ＊ -C(=O)NR ｂ -, -OC(=O)O-, ＊ -NR ａ C(=O)O-, ＊ -OC(=O)NR ｂ - ＊ -NR ａ C(=O)NR ｂ - ＊ -OC(=S)- ＊ -C(=S)O- ＊ -NR ａ C(=S)- ＊ -C(=S)NR ｂ -, -OC(=S)O-, ＊ -NR ａ C(=S)O-, ＊ -OC(=S)NR ｂ - ＊ -NR ａ C(=S)NR ｂ - ＊ -OC(=O)S-, ＊ -SC(=O)O-, ＊ -NR ａ C(=O)S- ＊ -SC(=O)NR ｂ - ＊ -OS(O) ｘ - ＊ -S(O) ｘ O-, ＊ -NR ａ S(O) ｘ -, or ＊ -S(O) ｘ NR ｂ - and ＊ This points in the direction toward R. R'' is a monocyclic, condensed, crosslinked, or spirocyclic portion, and the ring portion is one or more C １ ~C ６ It is optionally substituted with alkyl, oxo, or phenyl, and the alkyl or phenyl is optionally substituted with one or more halogens, phenyl, or phenoxy, and the phenyl or phenoxy is optionally substituted with one or more halogens or C １ ~C ６ It is optionally replaced with an alkoxy, (ii) R is C １２ ~C ３２ It is alkenyl, (iii)m is 1 or 2, and L is -O- or ＊ -C(=O)NR ｂ - and or (iv)m is 1 or 2, and at least one G is C ３ ~C １０ A cycloalkylene compound, or its stereoisomer, or a pharmaceutically acceptable salt thereof.

6. Z ２ The compound according to claim 5, wherein is -OC(=O)NH-, -NHC(=O)-, -NHC(=O)NH-, or -NHC(=O)S-.

7. X is O, Y is O, Z ２ If R is -O-, ２ is, -(C ２ ~C ６ Alkylene)-C(=O)NH-R or-(C ２ ~C ６ The compound according to claim 5, wherein it is alkylene)-O-R.

8. A compound of formula (IV), 【Chemistry 15】 Here, Each example in G' is independent of C １ ~C ８ Alkylene or C ３ ~C ８ It is a cycloalkylene, Each example in L' is independent of the others. ＊ -NR ｃ C (= O) - ＊ This indicates the direction toward the pyrrolidine-2,5-dione ring. R ｃ Each example is independently H, C １ ~C ３２ Alkyl, or C ２ ~C ３２ It is alkenyl, R is C ４ ~C ３２ Alkyl or C ４ ~C ３２ It is alkenyl, A compound in which m is 0, 1, or 2. or its stereoisomer, or a pharmaceutically acceptable salt thereof.

9. R is -L"-(CH ２ ) ０－３ It is replaced with -R, and here, L represents non-existence, -O-, -NR ａ -, -C(=O)-, ＊ -OC (=O)- ＊ -C(=O)O- ＊ -NR ａ C(=O)- ＊ -C(=O)NR ｂ -, -OC(=O)O-, ＊ -NR ａ C(=O)O-, ＊ -OC(=O)NR ｂ - ＊ -NR ａ C(=O)NR ｂ - ＊ -OC(=S)- ＊ -C(=S)O- ＊ -NR ａ C(=S)- ＊ -C(=S)NR ｂ -, -OC(=S)O-, ＊ -NR ａ C(=S)O-, ＊ -OC(=S)NR ｂ - ＊ -NR ａ C(=S)NR ｂ - ＊ -OC(=O)S-, ＊ -SC(=O)O-, ＊ -NR ａ C(=O)S- ＊ -SC(=O)NR ｂ - ＊ -OS(O) ｘ - ＊ -S(O) ｘ O-, ＊ -NR ａ S(O) ｘ -, or ＊ -S(O) ｘ NR ｂ - and ＊ This points in the direction toward R. R ａ Each of these examples is independently H or C １ ~C ６ It is alkyl, R ｂ Each of these examples is independently H or C １ ~C ６ It is alkyl, x is either 1 or 2, R'' is a monocyclic, condensed, crosslinked, or spirocyclic portion, and the ring portion is one or more C １ ~C ６ It is optionally substituted with alkyl, oxo, or phenyl, and the alkyl or phenyl is optionally substituted with one or more halogens, phenyl, or phenoxy, and the phenyl or phenoxy is optionally substituted with one or more halogens or C １ ~C ６ The compound according to claim 1, which is optionally substituted with an alkoxy.

10. An oligonucleotide comprising at least one lipophilic monomer of the following formula, 【Chemistry 19】 In equations (V) and (VII), X is either O or S, Y is O or NR a, D1 is H, a hydroxyl protecting group, or a reactive phosphorus group. D2 is H, a hydroxyl protecting group, or a reactive phosphorus group. Alternatively, D1 and D2 together form protecting groups for both the oxygen and Y to which they are bonded. Z1, Z2, and Z3 are independently -O-, -NR a-, * -OC(=O)-, * -NR a C(=O)-, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b-, * -NR a C(=O)NR b-, * -OC(=S)-, * -NR a C(=S)-, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b-, * -NR a C(=S)NR b-, * -OC(=O)S-, * -NR a C(=O)S-, * -OS(O) x or * -NR a S(O) x -, where * indicates the direction toward the ring containing X, R1, R2, and R3 are each independently H, a C1-C6 alkyl optionally substituted with a C1-C6 alkoxy, or -(G-L)m-R, wherein at least one of R1, R2, and R3 is -(G-L)m-R. Each example in G is independently a C1-C8 alkylene. Each example of L is independent of the following: -O-, -NR a-, -C(=O)-, * -OC(=O)-, * -C(=O)O-, * -NR a C(=O)-, * -C(=O)NR b-, -OC(=O)O-, * -NR a C(=O)O-, * -OC(=O)NR b-, * -NR a C(=O)NR b-, * -OC(=S)-, * -C(=S)O-, * -NR a C(=S)-, * -C(=S)NR b-, -OC(=S)O-, * -NR a C(=S)O-, * -OC(=S)NR b-, * -NR a C(=S)NR b-, *-OC(=O)S-, *-SC(=O)O-, *-NR a C(=O)S-, *-SC(=O)NR b-, *-OS(O) x-, *-S(O) x O-, *-NR a S(O) x-, or *-S(O) x NR b-, where * indicates the direction toward the ring containing X, Each example of Ra is independently H or C1-C6 alkyl. Each example of R b is independently H or C1-C6 alkyl, Each example of R is independently a C12-C32 alkyl or C12-C32 alkenyl. x is either 1 or 2, n is either 0 or 1, m is 0, 1, or 2; In equation (VIII), Each example in G' is independently a C1-C8 alkylene or a C3-C8 cycloalkylene. Each example of L' is independently * -NR c C(=O)-, where * indicates the direction toward the pyrrolidine-2,5-dione ring. Each example of R c is independently H, C1-C32 alkyl, or C2-C32 alkenyl. R is a C4-C32 alkyl or C4-C32 alkenyl, An oligonucleotide in which m is 0, 1, or 2.

11. The oligonucleotide according to claim 10, wherein the oligonucleotide is an antisense, antagonist, microRNA, siRNA, premicroRNA, antimyr, ribozyme, RNA activator, U1 adapter, immunostimulator, or aptamer.

12. The oligonucleotide according to claim 10, further comprising a targeted ligand.

13. An oligonucleotide according to claim 10 for delivery to cells, wherein the oligonucleotide comprises at least one lipophilic monomer of formula (V).

14. An oligonucleotide according to claim 10, used in a method for reducing the expression of a target gene in cells, wherein the method comprises contacting the cells with the oligonucleotide, the oligonucleotide comprising at least one lipophilic monomer of formula (V).

15. An oligonucleotide according to claim 10 for delivery to cells, wherein the oligonucleotide comprises at least one lipophilic monomer of formula (VII).

16. An oligonucleotide according to claim 10 for delivery to cells, wherein the oligonucleotide comprises at least one lipophilic monomer of formula (VIII).

17. An oligonucleotide according to claim 10, used in a method for reducing the expression of a target gene in cells, wherein the method comprises contacting the cells with the oligonucleotide, the oligonucleotide comprising at least one lipophilic monomer of formula (VII).

18. An oligonucleotide according to claim 10, used in a method for reducing the expression of a target gene in cells, wherein the method comprises contacting the cells with the oligonucleotide, the oligonucleotide comprising at least one lipophilic monomer of formula (VIII).

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