Lipid monomers for therapeutic delivery of RNA

Abstract

Provided herein are compounds comprising one or more lipophilic moieties and capable of being used to prepare one or more lipophilic monomers. Also provided herein are lipophilic monomers capable of being conjugated to one or more positions on at least one strand of an oligonucleotide, and oligonucleotides comprising at least one of the lipophilic monomers.

Description

Background Art

[0001] The effectiveness of delivery of therapeutic RNA outside the liver is a fundamental obstacle to its clinical application. Conjugate-mediated delivery is becoming the main clinical delivery paradigm for siRNA. Lipids are the main types of conjugates widely used to improve siRNA delivery, because lipid conjugation increases the plasma half-life and enhances the tissue accumulation and cellular uptake of siRNA. siRNA is highly hydrophilic and has poor pharmacological properties. Lipid conjugation regulates the hydrophobicity of siRNA, which controls pharmacokinetic behavior and tissue biodistribution by driving selectivity and in situ incorporation into endogenous lipoprotein pathways. Although siRNA has been conjugated to fatty acids, cholesterol, and tocopherol, there is still a continuous demand for the development of effective and robust lipid delivery systems that can balance safety and efficacy to achieve a better therapeutic range. The present application relates to siRNA conjugated to a variety of lipid types for delivering siRNA to other tissues outside the liver. Summary of the invention

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

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

[0004]

[0005] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein X, Y, D 1 , D 2 , Z 1 , Z 2 , Z 3 , R 1 , R 2 , R 3 and n are as defined herein or elsewhere.

[0006] In one embodiment, provided herein is a compound of formula (III):

[0007]

[0008] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein X, Y, D 1 , D 2 , Z 2 , R 2 and B are as defined herein or elsewhere.

[0009] In one embodiment, provided herein is a compound of formula (IV):

[0010]

[0011] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein G', L', R and m are as defined herein or elsewhere.

[0012] In one embodiment, provided herein is an oligonucleotide comprising at least one lipophilic monomer of the formula:

[0013]

[0014] Where 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.

[0015] Also provided herein are methods of using the oligonucleotides provided herein to reduce the expression of a target gene in a cell or a subject. DETAILED DESCRIPTION

[0016] definition

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art. All patents, applications, published applications and other publications are incorporated by reference in their entirety. In the event that there are multiple definitions for a term in this article, the term in that section shall prevail unless otherwise stated.

[0018] As used herein and in the specification and 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.

[0019] As used herein, the terms "comprising" and "including" can be used interchangeably. The terms "comprising" and "including" should be interpreted as specifying the presence of the features or components mentioned, but not excluding the presence or addition of one or more features, components, or combinations thereof. In addition, the terms "comprising" and "including" are intended to include examples covered by the term "consisting of". Therefore, the term "consisting of" can be used instead of the terms "comprising" and "including" to provide more specific embodiments.

[0020] 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". An exception to this definition will only occur if a combination of elements, functions, steps, or actions are inherently mutually exclusive in some way.

[0021] As used herein, the phrase "and / or" as used in phrases such as "A and / or B" is intended herein to include both A and B; A or B; A (alone); and B (alone). Similarly, the phrase "and / or" as used in phrases such as "A, B, and / or C" is intended to encompass 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).

[0022] As used herein, and unless otherwise indicated, the term "polynucleotide" or "nucleic acid" as used interchangeably herein refers to a polymer of nucleotides of any length and includes, 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 synthesis reaction. Polynucleotides can include modified nucleotides, such as methylated nucleotides and their analogs. Nucleotides can be in single-stranded or double-stranded form. As used herein, and unless otherwise indicated, "nucleic acid" also includes nucleic acid mimics, such as locked nucleic acids (LNA), peptide nucleic acids (PNA) and morpholinos. As used herein, and unless otherwise indicated, the term "oligonucleotide" refers to a short synthetic polynucleotide, whose length is generally but not necessarily less than about 200 nucleotides. The terms "oligonucleotide" and "polynucleotide" are not mutually exclusive. The above description of polynucleotides is equally and fully applicable to oligonucleotides. Unless otherwise specified, the left end of any single-stranded polynucleotide sequence disclosed herein is the 5' end; the left direction of a double-stranded polynucleotide sequence is referred to as the 5' direction. The 5' to 3' direction of addition of a nascent RNA transcript is referred to as the transcription direction; a sequence region on a DNA chain having the same sequence as an RNA transcript, which is at the 5' to 5' end of the RNA transcript, is referred to as an "upstream sequence"; a sequence region on a DNA chain having the same sequence as an RNA transcript, which is at the 3' to 3' end of the RNA transcript, is referred to as a "downstream sequence".

[0023] As used herein, the term "double-stranded RNA agent" or "dsRNA agent" means an agent containing RNA or RNA-like (e.g., chemically modified RNA) oligonucleotide molecules that can inhibit gene expression in a sequence-specific manner. The sense strand and / or antisense strand of a dsRNA agent may contain another portion (e.g., a lipid portion). For example, a lipid portion may be incorporated into the sense strand of a dsRNA agent. The sense strand and / or antisense strand of a dsRNA agent may be directly or indirectly connected or conjugated to another portion (e.g., a lipid portion). For example, the sense strand of a dsRNA agent may be directly or indirectly connected or conjugated to another portion (e.g., a lipid portion). In a specific embodiment, 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 forming 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 less 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 fully complementary, partially complementary, or substantially complementary to the sense strand. In a specific embodiment, the antisense strand of the dsRNA agent is partially complementary to the target RNA transcript. In another specific embodiment, the antisense strand of the dsRNA agent is substantially complementary to the target RNA transcript. In another specific embodiment, the antisense strand of the dsRNA agent is fully complementary to the target RNA transcript.

[0024] The two chains forming the double-stranded region or duplex structure can be different parts of a larger RNA molecule, or they can be separate RNA molecules. When the two chains are part of a larger RNA molecule, the continuous nucleotide chain between the 3' end of one chain and the 5' end of the corresponding other chain is connected to form a double-stranded region or duplex structure, and the connected RNA chain is referred to as a "hairpin loop". The hairpin loop can include at least one unpaired nucleotide. In some embodiments, the hairpin loop can include 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 that are not directed to the target site of the dsRNA agent. In some embodiments, the hairpin loop can be 1 to 10 unpaired nucleotides. In some embodiments, the hairpin loop can be 1 to 8 unpaired nucleotides. In some embodiments, the hairpin loop can be 4 to 10 unpaired nucleotides. In some embodiments, the hairpin loop can be 4 to 8 unpaired nucleotides.

[0025] When the two substantially complementary strands of a dsRNA agent consist of separate RNA molecules, these molecules need not, but may be covalently linked. In certain embodiments, where the two strands are covalently linked by a continuous chain of nucleotides between the 3' end of one strand and the 5' end of the corresponding other strand that forms a duplex structure, the linking structure is referred to as a "linker". The RNA strands of a dsRNA agent may have the same or different numbers of nucleotides.

[0026] In some embodiments, one or both strands of the dsRNA agent comprise an overhang. In other embodiments, the dsRNA agent is blunt-ended.

[0027] In a specific embodiment, the dsRNA agents described herein mediate the inhibition of messenger RNA (mRNA) degradation or mRNA translation in a sequence-specific manner. In a specific embodiment, the dsRNA agents described herein inhibit gene expression through the RNA-induced silencing complex (RISC) approach. In some embodiments, without being bound by theory, the functions of the dsRNA agents described herein are similar to short interfering RNA (siRNA). In a specific embodiment, the dsRNA agents described herein are siRNA.

[0028] As used herein, the term "complementary" when used to describe the relationship between a first nucleotide sequence (e.g., the sense strand of a dsRNA agent or a target mRNA) and a second nucleotide sequence (e.g., the antisense strand of a dsRNA agent or a single-stranded antisense oligonucleotide) means the ability of an oligonucleotide or polynucleotide comprising the first nucleotide sequence to hybridize (form base pair hydrogen bonds under mammalian physiological conditions (or similar conditions in vitro)) and form a duplex or double helix structure under certain conditions with an oligonucleotide or polynucleotide comprising the second nucleotide sequence. Complementary sequences include Watson Crick base pairs or non-Watson Crick base pairs, and include natural or modified nucleotides or nucleotide mimetics at least to the extent that the above hybridization requirements are met. Sequence identity or complementarity is independent of modification. For example, for the purpose of determining identity or complementarity, fA and mA are complementary to U (or T) and are identical to A.

[0029] As used herein, the term "fully complementary" in the context of two nucleotide sequences means that all (100%) bases in the contiguous sequence of the first nucleotide sequence will hybridize with the same number of bases in the contiguous sequence of the second nucleotide sequence to form a duplex. When two nucleotide sequences are designed to form one or more single-stranded overhangs after hybridization, such overhangs should not be considered mismatches in terms of the determination of complementarity. For example, for the purposes described herein, a dsRNA agent comprising an oligonucleotide of 21 nucleotides in length and another oligonucleotide of 23 nucleotides in length is considered "fully complementary", wherein the 23-nucleotide oligonucleotide comprises a sequence of 21 nucleotides that is fully complementary to the 21-nucleotide oligonucleotide. In a specific embodiment, the two nucleotide sequences are "fully complementary" when all (100%) bases in the first nucleotide sequence hybridize with all (100%) bases in the second nucleotide sequence to form a duplex. In a specific embodiment, the two nucleotide sequences hybridize under stringent conditions. In another specific embodiment, the two nucleotide sequences hybridize under very stringent conditions.

[0030] As used herein, the term "partial complementarity" in the context of two nucleotide sequences means that at least 65% but less than 80% of the bases in the contiguous sequence of the first nucleotide sequence will hybridize with the same number of bases in the contiguous sequence of the second nucleotide sequence to form a duplex. In a specific embodiment, the two nucleotide sequences hybridize under stringent conditions. In another specific embodiment, the two nucleotide sequences hybridize under very stringent conditions.

[0031] 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 continuous sequence of the first nucleotide sequence will hybridize with the same number of bases in the continuous sequence of the second nucleotide sequence to form a duplex. In some embodiments, 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 continuous sequence of the first oligonucleotide will hybridize with the same number of bases in the continuous sequence of the second oligonucleotide to form a duplex, the two nucleotide sequences are substantially complementary. In a specific embodiment, the two nucleotide sequences hybridize under stringent conditions. In another specific embodiment, the two nucleotide sequences hybridize under very stringent conditions.

[0032] As used herein, when referring to a numerical value, the terms "about" and "approximately" encompass the numerical value and a variation within + / - 20%. For example, about 20% would encompass values ​​between 16% and 24%, including 20%. In one embodiment, when referring to a numerical value, the terms "about" and "approximately" encompass the numerical value and a variation within + / - 15%. In another embodiment, when referring to a numerical value, the terms "about" and "approximately" encompass the numerical value and a variation within + / - 10%. In another embodiment, when referring to a numerical value, the terms "about" and "approximately" encompass the numerical value and a variation within + / - 5%.

[0033] As used herein, when referring to hybridization, the term "stringent" means that under "stringent conditions" or "stringent hybridization conditions", a first nucleotide sequence will hybridize to a second nucleotide with minimal hybridization to other sequences. In a specific embodiment, the antisense sequence will hybridize to its target sequence under stringent conditions with minimal targeting to other sequences. Stringent conditions depend on the sequence (e.g., sequence length, complementarity) 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, 50°C or 70°C for 12-16 hours, followed by washing. Those skilled in the art will appreciate that variations in hybridization stringency are inherently described.

[0034] As used herein, when it comes to hybridization, the term "very stringent" means that under "very stringent conditions" or "very stringent hybridization conditions", only the first nucleotide sequence will be observed to hybridize with the second nucleotide. In a specific embodiment, it will be observed that the antisense sequence hybridizes with its target sequence only under very stringent conditions. In addition, very stringent conditions may not allow hybridization between partially complementary sequences. Very stringent conditions depend on the sequence (e.g., sequence length, complementarity) and vary under different environmental parameters (e.g., assay conditions, physiological environment). Compared with stringent conditions under the same circumstances, very stringent conditions can include higher temperatures, lower ionic strengths and / or shorter reaction times. For example, very 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. It will be understood by those skilled in the art that the variation of hybridization stringency is inherently described.

[0035] As used herein, "target sequence" refers to a continuous portion of the nucleotide sequence of an RNA molecule formed during gene transcription, including mRNA (e.g., mRNA produced by alternative splicing) as an RNA processing product of a primary transcription product. In one embodiment, the continuous portion of the nucleotide sequence is at least long enough to serve as a substrate for RNAi-guided cleavage at or near the nucleotide sequence portion of the mRNA molecule formed during gene transcription. In a specific embodiment, the length of the target sequence is about 15-30 nucleotides. For example, the target sequence can 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, 19-30, 19-31, 19-32, 19-33, 19-34, 19-35, 19-36, 19-37, 19-38, 19-39, 19-40, 19-41, 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 In some 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 ranges and lengths are also considered part of the present disclosure.

[0036] As used herein, the phrases "a nucleotide sequence corresponding to any antisense strand nucleotide sequence", "a nucleotide sequence corresponding to an antisense strand nucleotide sequence", "a nucleotide sequence corresponding to any sense strand nucleotide sequence", "a nucleotide sequence corresponding to a sense strand nucleotide sequence", or "a nucleotide sequence corresponding to any nucleotide sequence" refer to an oligonucleotide comprising a nucleotide chain comprising the unmodified nucleotide, or one or more modified nucleotides, or one or more conjugated moieties (e.g., moieties described herein, such as lipids, or modified nucleotides conjugated to moieties described herein). It is known to those skilled in the art that the unmodified nucleotides may be replaced by other moieties without substantially changing the base pairing properties of the oligonucleotide comprising the nucleotide with such replacement moieties. For example, a nucleotide comprising inosine as its base may base pair with a nucleotide containing adenine, cytosine, or uracil. Thus, a nucleotide containing uracil, guanine, or adenine may be replaced by a nucleotide containing, for example, inosine. In another example, adenine and cytosine may be replaced by guanine and uracil, respectively, to form a GU wobble base pair with a target mRNA.

[0037] As used herein, the phrase "non-naturally occurring double-stranded ribonucleic acid (dsRNA) agent," "non-naturally occurring double-stranded ribonucleic acid agent," or "non-naturally occurring dsRNA agent" refers to a dsRNA agent that is not found in nature. Non-naturally occurring dsRNAs may contain one or more modified nucleotides.

[0038] As used herein, the term "overhang" in the context of a 5' or 3' nucleotide overhang refers to at least one unpaired nucleotide that protrudes from the duplex structure of a dsRNA agent. For example, when the 3' end of one strand of a dsRNA agent extends beyond the 5' end of the other strand, or vice versa, there is a nucleotide overhang. 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 in the antisense strand. In another embodiment, the 3' overhang is present in 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 in the antisense strand. In another embodiment, the 5' overhang is present in the sense strand. The overhang may be due to one strand being longer than the other, or being 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. The nucleotides in the overhang region of the dsRNA agent can each independently be a modified or unmodified nucleotide (eg, a 2'-fluoro-modified nucleotide, a 2'-O-methyl modified nucleotide, a deoxynucleotide, or a combination thereof).

[0039] In some embodiments, the 5' or 3' overhang of the sense strand or antisense strand of the dsRNA agent is phosphorylated. In certain embodiments, the 5' or 3' overhang of the sense strand and antisense strand of the dsRNA agent is phosphorylated. In some embodiments, the overhang region contains two (or more) nucleotides, with a phosphorothioate between the two (or more) nucleotides, and the two (or more) nucleotides can be the same or different.

[0040] In some embodiments, the dsRNA agent contains only a single overhang, which can enhance the interference activity of the 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 the dsRNA agent, or alternatively, at the 3' end of the antisense strand of the dsRNA agent. The dsRNA agent can also have a flat 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 the dsRNA agent has a nucleotide overhang at the 3' end, and the 5' end is a flat end.

[0041] In some embodiments, one strand of the dsRNA agent comprises at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides of the 5' end, 3' end, or both the 5' end and the 3' end overhang. In certain embodiments, one strand of the dsRNA agent comprises at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides but no more than 5 nucleotides of the 5' end, 3' end, or both the 5' end and the 3' end overhang. In some embodiments, one strand of the dsRNA agent comprises 1 nucleotide, 2 nucleotides, or 3 nucleotides of the 5' end, 3' end, or both the 5' end and the 3' end overhang. In certain embodiments, one strand of the dsRNA agent comprises 1 to 2 nucleotides, 1 to 3 nucleotides, 1 to 4 nucleotides, or 1 to 5 nucleotides of the 5' end, 3' end, or both the 5' end and the 3' end overhang. In some embodiments, one strand of the dsRNA agent comprises 2 to 3 nucleotides, 2 to 4 nucleotides, or 2 to 5 nucleotides of the 5' end, 3' end, or both the 5' end and the 3' end overhang. In certain embodiments, one strand of the dsRNA agent comprises a 5' end, a 3' end, or both a 5' end and a 3' end overhang of 3 to 4 nucleotides or 4 to 5 nucleotides. The strand may be an antisense strand or a sense strand. The nucleotide overhang may comprise or consist of a nucleotide analog or a nucleoside analog.

[0042] In some embodiments, each strand of the dsRNA agent comprises at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides of 5' end, 3' end, or both 5' end and 3' end overhangs. In certain embodiments, each strand of the dsRNA agent comprises at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides of 5' end, 3' end, or both 5' end and 3' end overhangs of at least 1 nucleotide, at least 2 nucleotides, or at least 3 nucleotides but no more than 5 nucleotides. In some embodiments, each strand of the dsRNA agent comprises 1 nucleotide, 2 nucleotides, or 3 nucleotides of 5' end, 3' end, or both 5' end and 3' end overhangs. In certain embodiments, each strand of the dsRNA agent comprises 1 to 2 nucleotides, 1 to 3 nucleotides, 1 to 4 nucleotides, or 1 to 5 nucleotides of 5' end, 3' end, or both 5' end and 3' end overhangs. In some embodiments, each strand of the dsRNA agent comprises 2 to 3 nucleotides, 2 to 4 nucleotides, or 2 to 5 nucleotides at the 5' end, 3' end, or both the 5' end and the 3' end overhang. In certain embodiments, each strand of the dsRNA agent comprises 3 to 4 nucleotides or 4 to 5 nucleotides at the 5' end, 3' end, or both the 5' end and the 3' end overhang. The nucleotide overhangs may comprise or consist of nucleotide analogs or nucleoside analogs.

[0043] As used herein, in the context of dsRNA agents, the term "blunt end" or "blunt-ended" refers to the absence of unpaired nucleotides or nucleotide analogs at a given end of the dsRNA, i.e., 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 include 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.

[0044] As used herein, in the context of a dsRNA agent, the term "antisense strand" or "guide strand" refers to the strand that contains the region that is complementary to the target sequence.

[0045] As used herein, in the context of a dsRNA agent, the term "sense strand" or "passenger strand" refers to the strand of the dsRNA agent that includes a region that is complementary to a region of the antisense strand.

[0046] As used herein, in the context of nucleobases of dsRNA agents, the term "modified" refers to nucleobases that do not naturally occur in RNA molecules. Naturally occurring RNA sequences include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U).

[0047] As used herein, in the context of nucleotides of an RNA sequence, such as a dsRNA agent, the term "modified" refers to nucleotides that are not naturally present in RNA molecules.

[0048] As used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the US Pharmacopeia, the European Pharmacopeia, or other generally recognized pharmacopoeia for use in animals, and more particularly in humans.

[0049] In one embodiment, each component is "pharmaceutically acceptable" in the sense of being compatible with the other ingredients of the pharmaceutical composition and suitable for use in contact with tissues or organs of humans and animals without excessive toxicity, irritation, allergic response, immunogenicity, or other problems or complications 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, a pharmaceutically acceptable excipient is a pH buffered aqueous solution.

[0050] As used herein, and unless otherwise specified, the term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable, relatively non-toxic acids, including inorganic 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, dihydrogenphosphoric acid, vinylsulfonic 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, monohydrogenphosphoric acid, 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., SM Berge et al., J. Pharm. Sci., 66: 1-19 (1977); and Handbook of Pharmaceutical Salts: Properties, Selection and Use, PH Stahl and CG Wermuth, eds., (2002), Wiley, Weinheim). In certain embodiments, suitable acid is a strong acid (e.g., pKa is 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 nontoxic compounds with acidic characteristics, including amino acids, such as aspartic acid, etc., and other compounds, such as aspirin, ibuprofen, saccharin, etc. Acid addition salts can be obtained by contacting the neutral form of the compound with a sufficient amount of the desired acid (pure 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.

[0051] As used herein, and unless otherwise indicated, the term "protecting group" refers to a chemical group that blocks reactive functional groups in a compound (such as, but not limited to, carboxyl, hydroxyl, and amino moieties) from undesired reactions. Protecting groups known to those skilled in the art are provided herein, such as those described in Protective Groups in Organic Synthesis, Greene, TW; Wuts, PGM, John Wiley & Sons, New York, NY, (5th edition, 2014), which can be added or removed using the procedures described therein. Examples of protected hydroxyl groups include, but are not limited to, silyl ethers, such as those obtained by reacting a hydroxyl group with a reagent such as, but not limited to, tert-butyldiphenylchlorosilane, tert-butyldimethylchlorosilane, trimethylchlorosilane, triisopropylchlorosilane, triethylchlorosilane; substituted methyl and ethyl ethers such as, but not limited to, methoxymethyl ether, methylthiomethyl ether, benzyloxymethyl ether, tert-butoxymethyl ether, 2-methoxyethoxymethyl ether, tetrahydropyranyl ether, 1-ethoxyethyl ether, allyl ether, benzyl ether, 2-cyanoethyl ether; esters such as, but not limited to, benzoylformate, formates, acetates, trichloroacetates, and trifluoroacetates. Examples of protecting groups include, but are not limited to, amidines (e.g., Me2N-CH=).

[0052] As used herein, and unless otherwise indicated, the term "reactive phosphorus group" refers to a chemical group or moiety that can be used to form a bond between nucleosides, including, for example, phosphodiester and thiophosphate nucleoside bonds. Such reactive phosphorus groups are known in the art and contain phosphorus atoms of P III or PV valence states, including but not limited to phosphoramidites, H-phosphonates, phosphotriesters, and phosphorus-containing chiral auxiliaries. In one embodiment, solid phase synthesis utilizes phosphoramidites (P III chemistry) as reactive phosphites. Subsequently, known methods are used to oxidize intermediate phosphite compounds to PV states, to produce phosphodiester or thiophosphate nucleoside bonds in some embodiments.

[0053] As used herein, and unless otherwise indicated, the term "internucleoside bond" or "internucleoside linking group" is intended to include all forms 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 acetyl and methyleneimino, and neutral non-ionic internucleoside linking groups such as 3'-CH2-C(═O)-N(H)-5' or 3'-CH2-N(H)-C(═O)-5'.

[0054] As used herein, and unless otherwise specified, the term "alkyl" refers to a saturated straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms. In one embodiment, an alkyl group has, for example, one to forty carbon atoms (C1-C 40 Alkyl), one to twenty-four carbon atoms (C1-C 24 Alkyl), four to twenty carbon atoms (C4-C 20 Alkyl), six to sixteen carbon atoms (C6-C 16 alkyl), six to nine carbon atoms (C6-C9 alkyl), one to fifteen carbon atoms (C1-C 15 Alkyl), one to twelve carbon atoms (C1-C 12 The term "alkyl" refers to a group of carbon atoms (C1-C8 alkyl), one to eight carbon atoms (C1-C8 alkyl), or one to six carbon atoms (C1-C6 alkyl) and is 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 (tert-butyl), 3-methylhexyl, 2-methylhexyl, pentadecyl, hexadecyl, (1s,3r,5R,7S)-1-heptyl-3-octyl adamantane, and the like. Unless otherwise specified, alkyl groups are optionally substituted.

[0055] As used herein, and unless otherwise indicated, the term "alkenyl" refers to a straight or branched hydrocarbon chain group consisting only of carbon and hydrogen atoms, containing one or more carbon-carbon double bonds. As understood by those of ordinary skill in the art, the term "alkenyl" also includes groups with "cis" and "trans" configurations, or alternatively "E" and "Z" configurations. In one embodiment, an alkenyl group has, for example, two to forty carbon atoms (C2-C 40 alkenyl), two to twenty-four carbon atoms (C2-C 24 alkenyl), four to twenty carbon atoms (C4-C 20 alkenyl), six to sixteen carbon atoms (C6-C 16 alkenyl), six to nine carbon atoms (C6-C9 alkenyl), two to fifteen carbon atoms (C2-C 15 alkenyl), two to twelve carbon atoms (C2-C 12 The term "alkenyl" refers to a group of 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 is attached to the rest of the molecule by a single bond. Examples of alkenyl groups include, but are not limited to, vinyl, prop-1-enyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, (4Z,7Z,10Z,13Z)-nonadeca-4,7,10,13-tetraene, (4Z,7Z,10Z,13Z)-16,16-dimethyleicosah-4,7,10,13-tetraene, and the like. Unless otherwise specified, alkenyl groups are optionally substituted.

[0056] As used herein, and unless otherwise specified, the term "alkylene" or "alkylene chain" refers to a saturated straight or branched multivalent (e.g., divalent or trivalent) hydrocarbon chain consisting solely of carbon and hydrogen that connects the remainder of the molecule to a free radical (group). In one embodiment, an alkylene group has, for example, one to twenty-four carbon atoms (C1-C 24 Alkylene), one to fifteen carbon atoms (C1-C 15 Alkylene), one to twelve carbon atoms (C1-C 12 The alkylene group may be a 1-4 carbon atom (C1-C8 alkylene), a 1-6 carbon atom (C1-C6 alkylene), a 2-4 carbon atom (C2-C4 alkylene), a 1-2 carbon atom (C1-C2 alkylene). 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 by a single bond and to the free radical by a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the free radical may be through one carbon or any two (or more) carbons within the chain. Unless otherwise specified, an alkylene chain is optionally substituted.

[0057] As used herein, and unless otherwise indicated, the term "alkenylene" refers to a straight or branched multivalent (e.g., divalent or trivalent) hydrocarbon chain consisting solely of carbon and hydrogen that connects the remainder of the molecule to a free radical (group), containing one or more carbon-carbon double bonds. In one embodiment, an alkenylene group has, for example, two to twenty-four carbon atoms (C2-C 24 alkenyl), two to fifteen carbon atoms (C2-C 15 alkenyl), two to twelve carbon atoms (C2-C 12 Alkenylene is a 1- to 2-carbon atom (C-C alkenylene), 2 to 8 carbon atoms (C-C alkenylene), 2 to 6 carbon atoms (C-C alkenylene), or 2 to 4 carbon atoms (C-C alkenylene). Examples of alkenylene include, but are not limited to, vinylene, propenylene, n-butenylene, and the like. Alkenylene is attached to the rest of the molecule by a single or double bond and to the free radical by a single or double bond. The point of attachment of an alkenylene to the rest of the molecule and to the free radical can be through one carbon or any two (or more) carbons within the chain. Unless otherwise indicated, alkenylene is optionally substituted.

[0058] As used herein, and unless otherwise indicated, the term "cycloalkyl" refers to a non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms, and is saturated. Cycloalkyl groups may include fused rings, bridged rings, or spiro ring systems. In one embodiment, the cycloalkyl group has, for example, 3 to 15 ring carbon atoms (C3-C 15 cycloalkyl), 3 to 10 ring carbon atoms (C3-C 10Cycloalkyl) or 3 to 8 ring carbon atoms (C3-C8 cycloalkyl). The cycloalkyl is attached to the rest of the molecule by a single bond. Examples of monocyclic cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyls include, but are not limited to, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, and the like. Unless otherwise stated, when a cycloalkyl is fused to an aromatic ring, the resulting fused ring is still considered to be a cycloalkyl. Unless otherwise stated, a cycloalkyl group is optionally substituted.

[0059] As used herein, and unless otherwise specified, the term "cycloalkylene" is a multivalent (eg, divalent or trivalent) cycloalkyl group. Unless otherwise specified, a cycloalkylene group is optionally substituted.

[0060] As used herein, and unless otherwise indicated, the term "heterocyclyl" refers to a non-aromatic monocyclic or polycyclic moiety containing one or more (e.g., one, one or two, one to three or one to four) heteroatoms independently selected from nitrogen, oxygen, phosphorus and sulfur. The heterocyclyl group can be attached to the main structure at any heteroatom or carbon atom. The heterocyclyl group can be a monocyclic, bicyclic, tricyclic, tetracyclic or other polycyclic ring system, wherein the polycyclic ring system can be a fused ring, a bridged ring or a spiro ring system. The heterocyclyl polycyclic ring system can include one or more heteroatoms in one or more rings. The heterocyclyl group can be saturated or partially unsaturated. Saturated heterocycloalkyl groups can be referred to as "heterocycloalkyl". If the heterocyclyl contains at least one double bond, the partially unsaturated heterocycloalkyl group can be referred to as "heterocycloalkenyl", or if the heterocyclyl contains at least one triple bond, it is referred to as "heterocycloalkynyl". In one embodiment, the heterocyclyl group 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). Whenever it appears in this article, a numerical range such as "3 to 18" refers to each integer in the given range; for example, "3 to 18 ring atoms" means that the heterocyclyl group can be composed 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 and including 18 ring atoms. Examples of heterocyclic groups include, but are not limited to, imidazolyl, imidazolidinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyrrolidinyl, furanyl, tetrahydrofuranyl, thiophenyl, pyridyl, piperidyl, quinolyl, and isoquinolyl. Unless otherwise indicated, heterocyclic groups are optionally substituted.

[0061] When groups described herein are referred to as "substituted", they may be substituted with any suitable substituent or substituents. 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, CI, 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', wherein: R' is independently H, C1-C 15 alkyl or cycloalkyl, and x is 0, 1 or 2. In some embodiments, the substituent is C1-C 12 In other embodiments, the substituent is a cycloalkyl group. In other embodiments, the substituent is a halo group, such as fluorine. In other embodiments, the substituent is a hydroxyl group. In other embodiments, the substituent is an alkoxy group (-OR'). In other embodiments, the substituent is a carboxyl group. In other embodiments, the substituent is an amino group (-NR'R').

[0062] As used herein, and unless otherwise indicated, the term "optional" or "optionally" (e.g., optionally substituted) means that the subsequently described event or circumstances may or may not occur, and that the description includes instances where the event or circumstances occurs and instances where the event or circumstances does not occur. For example, "optionally substituted alkyl" means that the alkyl may or may not be substituted, and that the description includes both substituted alkyls and alkyls having no substituents.

[0063] Compounds provided herein may contain one or more asymmetric centers, and thus may produce enantiomers, diastereomers, and other stereoisomeric forms, which may be defined as (R)- or (S)-, or (D)- or (L)- of amino acids according to absolute stereochemistry. Unless otherwise indicated, compounds provided herein are intended to include all of these 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 may be resolved using conventional techniques (e.g., chromatography and fractional crystallization). Conventional techniques for preparing / separating individual enantiomers include chiral synthesis of suitable optically pure precursors or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high pressure liquid chromatography (HPLC). When compounds described herein contain olefinic double bonds or other geometrically asymmetric centers, and unless otherwise indicated, the compounds are intended to include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0064] As used herein, unless otherwise specified, the term "isomer" refers to different compounds with the same molecular formula. "Stereoisomers" are isomers that differ only in the way their atoms are arranged in space. "Atropisomers" are stereoisomers that are hindered from rotating 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 ratio may be referred to as a "racemic" mixture. "Diastereomers" are stereoisomers that have at least two asymmetric atoms but are non-mirror images of each other.

[0065] "Stereoisomers" may also include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof. In certain embodiments, the compounds described herein are isolated as E or Z isomers. In other embodiments, the compounds described herein are mixtures of E and Z isomers.

[0066] It should be noted that if there is a discrepancy between the depicted structure and the name of that structure, the depicted structure will be given more weight.

[0067] Compound

[0068] Unless otherwise stated, the descriptions provided herein apply to all formulae provided herein (eg, formulae (I) to (VIII), including subformulae thereof), to the extent they are applicable.

[0069] In one embodiment, provided herein is a compound of formula (I):

[0070]

[0071] in:

[0072] X is O or S;

[0073] Y is O or NR a ;

[0074] D 1 is H, a hydroxyl protecting group or a reactive phosphorus group;

[0075] D 2 is H, a hydroxyl protecting group or a reactive phosphorus group;

[0076] or D 1 and D 2 Together they form protecting groups for both the oxygen to which they are attached and for Y;

[0077] Z 1 , Z 2 and Z 3 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 * refers to the direction toward the ring containing X;

[0078] R 1 , R 2 and R 3 Each is independently H, C1-C6 alkyl optionally substituted by C1-C6 alkoxy, or -(GL) m -R, condition is R 1 , R 2 and R 3 At least one of -(GL) m -R;

[0079] Each instance of G is independently C1-C8 alkylene;

[0080] 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 -; where * refers to the direction toward the ring containing X;

[0081] R a Each instance of is independently H or C1-C6 alkyl;

[0082] R b Each instance of is independently H or C1-C6 alkyl; and

[0083] Each instance of R is independently C 12 -C 32 Alkyl or C 12 -C 32 Alkenyl, or when R 1 , R 2 and R 3 More than one of -(GL) m -R, then one or more R is independently C 12 -C 32 Alkyl, C 12 -C 32alkenyl or optionally protected mannose;

[0084] x is 1 or 2;

[0085] n is 0 or 1;

[0086] m is 0, 1 or 2;

[0087] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0088] In one embodiment, X is O. In one embodiment, X is S.

[0089] In one embodiment, Y is O. In one embodiment, Y is NR a In one embodiment, Y is NH.

[0090] 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 (if present) are each independently -O-, -NH-, *-OC(=O)-, *-NHC(=O)-, -OC(=O)O-, *-NHC(=O)O-, *-OC(=O)NH-, or -NHC(=O)NH-. In one embodiment, Z 1 , Z 2 and Z 3 (if present) are -O-.

[0091] In one embodiment, n is 0. In one embodiment, the compound is of formula (IA):

[0092]

[0093] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0094] In one embodiment, the compound is of formula (IA-1):

[0095]

[0096] or a pharmaceutically acceptable salt thereof.

[0097] In one embodiment, the compound is of formula (II-A):

[0098]

[0099] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0100] In one embodiment, the compound is of formula (II-A-1):

[0101]

[0102] or a pharmaceutically acceptable salt thereof.

[0103] In one embodiment, n is 1. In one embodiment, the compound is of formula (IB):

[0104]

[0105] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0106] In one embodiment, the compound is of formula (IB-1):

[0107]

[0108] or a pharmaceutically acceptable salt thereof.

[0109] In one embodiment, the compound is of formula (II-B):

[0110]

[0111] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0112] In one embodiment, the compound is of formula (II-B-1):

[0113]

[0114] or a pharmaceutically acceptable salt thereof.

[0115] Unless otherwise indicated, when the stereochemistry of a position in a formula provided herein is not specified, the compound of the formula can be a 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 an α-isomer at the 1-position; in another embodiment, the compound is a β-isomer at the 1-position; and in yet another embodiment, the compound is a mixture of α-isomers and β-isomers.

[0116] In one embodiment, R 1 is H. In one embodiment, R 1 is a C1-C6 alkyl group. 1 In one embodiment, R 1 In one embodiment, R 1 In one embodiment, R 1 In one embodiment, R 1 In one embodiment, R 1 In one embodiment, R 1 In one embodiment, the alkyl group is substituted with a C1-C6 alkoxy group. In one embodiment, the alkoxy group is a methoxy group. In one embodiment, the alkoxy group is an ethoxy group. In one embodiment, the alkoxy group is a n-propoxy group. In one embodiment, R 1 It is 2-methoxyethyl (MOE).

[0117] In one embodiment, R 2 is H. In one embodiment, R 2 is a C1-C6 alkyl group. 2 In one embodiment, R 2 In one embodiment, R 2 In one embodiment, R 2 In one embodiment, R 2 In one embodiment, R 2 In one embodiment, R 2 In one embodiment, the alkyl group is substituted with a C1-C6 alkoxy group. In one embodiment, the alkoxy group is a methoxy group. In one embodiment, the alkoxy group is an ethoxy group. In one embodiment, the alkoxy group is a n-propoxy group. In one embodiment, R 2 It is 2-methoxyethyl (MOE).

[0118] In one embodiment, R 3is H. In one embodiment, R 3 is a C1-C6 alkyl group. 3 In one embodiment, R 3 In one embodiment, R 3 In one embodiment, R 3 In one embodiment, R 3 In one embodiment, R 3 In one embodiment, R 3 In one embodiment, the alkyl group is substituted with a C1-C6 alkoxy group. In one embodiment, the alkoxy group is a methoxy group. In one embodiment, the alkoxy group is an ethoxy group. In one embodiment, the alkoxy group is a n-propoxy group. In one embodiment, R 3 It is 2-methoxyethyl (MOE).

[0119] In one embodiment, R 1 -(GL) m -R. In one embodiment, R 2 -(GL) m -R. In one embodiment, R 3 -(GL) m -R. In one embodiment, R 1 and R 2 -(GL) m -R. In one embodiment, R 1 , R 2 and R 3 -(GL) m -R. In one embodiment, R 1 and R 2 -(GL) m -R, where R 1 R in is C 12 -C 32 Alkyl or C 12 -C 32 Alkenyl and R 2 In one embodiment, R is an optionally protected mannose. 1 and R 3 -(GL) m -R, where R 1 R in is C 12 -C 32 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 -(GL) m -R, where R 1 R in is C 12 -C 32 Alkyl or C 12 -C 32 Alkenyl and R 2 and R 3 R in which is optionally protected mannose. In some embodiments, the optionally protected mannose is protected by an acetyl moiety.

[0120] In one embodiment wherein n is 0, R 1 -(GL) m -R, and R 2 is a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 2 -(GL) m -R, and R 1 is a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 1 and R 2 Each independently is -(GL) m -R.

[0121] In one embodiment wherein n is 1, R 1 -(GL) m -R, and R 2 and R 3 Each is independently a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 2 -(GL) m -R, and R 1 and R 3 Each is independently a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 3 -(GL) m -R, and R 1 and R 2 Each is independently a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 1 and R 2 Each independently is -(GL) m -R, and R 3 is a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 1 and R 3 Each independently is -(GL)m -R, and R 2 is a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 2 and R 3 Each independently is -(GL) m -R, and R 1 is a C1-C6 alkyl group optionally substituted by a C1-C6 alkoxy group. 1 , R 2 and R 3 Each independently is -(GL) m -R.

[0122] In one embodiment, m is 0. In one embodiment, -(GL) m -R is -R.

[0123] In one embodiment, m is 1. In one embodiment, m is 2.

[0124] 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, the alkylene is C1 alkylene. In one embodiment, the alkylene is C2 alkylene. In one embodiment, the alkylene is C3 alkylene. In one embodiment, the alkylene is C4 alkylene. In one embodiment, the alkylene is C5 alkylene. In one embodiment, the alkylene is C6 alkylene. In one embodiment, the alkylene is -CH2-. In one embodiment, the alkylene is -CH2CH2-. In one embodiment, the alkylene is -CH2CH2CH2-.

[0125] 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 *-NHC(=O)NH-. In one embodiment, each instance of L is independently *-NHC(=O)-.

[0126] 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.

[0127] In one embodiment, R a is H. In one embodiment, R a is a C1-C6 alkyl group. a It is methyl.

[0128] In one embodiment, R b is H. In one embodiment, R b is a C1-C6 alkyl group. b It is methyl.

[0129] In one embodiment, provided herein is a compound of formula (III):

[0130]

[0131] in:

[0132] B is a modified or unmodified nucleobase;

[0133] X is O or S;

[0134] Y is O or NR a ;

[0135] D 1 is H, a hydroxyl protecting group or a reactive phosphorus group;

[0136] D 2 is H, a hydroxyl protecting group or a reactive phosphorus group;

[0137] or D 1 and D 2 Protecting groups for both the oxygen and Y to which they are attached;

[0138] 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 -; where * refers to the direction toward the ring containing X;

[0139] R 2 -(GL) m -R;

[0140] Each instance of G is independently C1-C8 alkylene or C3-C 10 Cycloalkylene;

[0141] 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 -; where * refers to the direction toward the ring containing X;

[0142] R a Each instance of is independently H or C1-C6 alkyl;

[0143] R b Each instance of is independently H or C1-C6 alkyl; and

[0144] R is C 12 -C 32 Alkyl or C 12 -C 32 alkenyl;

[0145] x is 1 or 2; and

[0146] m is 0, 1 or 2;

[0147] The condition is when X is O, Y is O and Z 2 When it is -O-,

[0148] (i) R is -L"-(CH2) 0-3 -R" where:

[0149] L" is absent, -O-, -NR a -, -C(=O)-,*-OC(=O)-,*-

[0150] C(=O)O-, *-NR a C(=O)-, *-C(=O)NR b -、-OC(=O)O-、*-

[0151] NR a C(=O)O-、*-OC(=O)NR b -、*-NR a C(=O)NR b -、*-OC(=S)-、*-

[0152] C(=S)O-, *-NR a C(=S)-, *-C(=S)NR b -、-OC(=S)O-、*-

[0153] NR a C(=S)O-、*-OC(=S)NR b -、*-NR a C(=S)NR b -、*-OC(=O)S-、*-

[0154] SC(=O)O-, *-NR a C(=O)S-、*-SC(=O)NR b -、*-OS(O) x -、*-

[0155] S(O) x O-, *-NR a S(O) x -or*-S(O) x NRb -; where * refers to the direction towards R;

[0156] and

[0157] R" is a monocyclic, fused, bridged or spirocyclic 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;

[0158] (ii) R is C 12 -C 32 alkenyl;

[0159] (iii) m is 1 or 2; and L is -O- or *-C(=O)NR b -;or

[0160] (iv) m is 1 or 2, and at least one G is C3-C 10 Cycloalkylene;

[0161] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0162] In one embodiment, X is O. In one embodiment, X is S.

[0163] In one embodiment, Y is O. In one embodiment, Y is NR a In one embodiment, Y is NH.

[0164] In one embodiment, the compound is of formula (III-A):

[0165]

[0166] or a pharmaceutically acceptable salt thereof.

[0167] In one embodiment, the compound is of formula (III-A-1):

[0168]

[0169] or a pharmaceutically acceptable salt thereof.

[0170] In one embodiment, Z 2 -O-, *-OC(=O)-, *-NR a C(=O)-, -OC(=O)O-, *-NR a C(=O)O-、*-OC(=O)NR b -、*-NR a C(=O)NRb -, *-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 For -O-.

[0171] In one embodiment, when X is O, Y is O and Z 2 When it is -O-, R is C 12 -C 32 In one embodiment, m is 0 and R is C 12 -C 32 Alkenyl.

[0172] In one embodiment, when X is O, Y is O and Z 2 When it is -O-, R is replaced by -L"-(CH2) 0-3 -R" is substituted. In one embodiment, m is 0 and R is -L"-(CH2) 0-3 -R". In one embodiment, m is 0, and R is substituted with a fused, bridged, or spirocyclic alkyl group. In one embodiment, R is substituted with adamantyl. In one embodiment, adamantyl is 1-adamantyl. In one embodiment, adamantyl is 2-adamantyl. In one embodiment, R is replace.

[0173] In one embodiment, when X is O, Y is O and Z 2 When it 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, when X is O, Y is O, and Z 2 When it is -O-, 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 -CH2CH2-C(=O)NH-R. In one embodiment, R 2In one embodiment, when X is O, Y is O, and Z is 2 When it is -O-, R 2 is -(C2-C6 alkylene)-OR. In one embodiment, R 2 is -(C2-C3 alkylene)-OR. In one embodiment, R 2 In one embodiment, R 2 It is -CH2CH2CH2-OR.

[0174] In one embodiment, when X is O, Y is O and Z 2 When -O-, m is 1 or 2, and at least one G is C3-C 10 In one embodiment, m is 1 and G is C3-C 10 Cycloalkylene.

[0175] In one embodiment, m is 0. In one embodiment, m is 1. In one embodiment, m is 2.

[0176] 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, the alkylene is C1 alkylene. In one embodiment, the alkylene is C2 alkylene. In one embodiment, the alkylene is C3 alkylene. In one embodiment, the alkylene is C4 alkylene. In one embodiment, the alkylene is C5 alkylene. In one embodiment, the alkylene is C6 alkylene. In one embodiment, the alkylene is -CH2-. In one embodiment, the alkylene is -CH2CH2-. In one embodiment, the alkylene is -CH2CH2CH2-.

[0177] In one embodiment, each instance of G is independently C3-C 10 Cycloalkylene. In one embodiment, each instance of G is independently C3-C8 cycloalkylene. In one embodiment, cycloalkylene is cyclopropylene. In one embodiment, cycloalkylene is cyclobutylene. In one embodiment, cycloalkylene is cyclopentylene. In one embodiment, cycloalkylene is cyclohexylene. In one embodiment, cycloalkylene is cycloheptylene. In one embodiment, cycloalkylene is cyclooctylene.

[0178] 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 *-NHC(=O)NH-. In one embodiment, each instance of L is independently *-C(=O)NH-.

[0179] 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.

[0180] In one embodiment, R a is H. In one embodiment, R a is a C1-C6 alkyl group. a It is methyl.

[0181] In one embodiment, R b is H. In one embodiment, R b is a C1-C6 alkyl group. b It is methyl.

[0182] 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, as provided herein or known in the art.

[0183] In one embodiment, D 1 is a hydroxyl protecting group. 1 is 4,4'-dimethoxytrityl chloride (DMTr). In one embodiment, D 1 It is 4-monomethoxytrityl (MMTr).

[0184] In one embodiment, D 2 is a reactive phosphorus group. 2 for

[0185] In one embodiment, D 1 H and D 2 For H.

[0186] In one embodiment, provided herein is a compound of formula (IV):

[0187]

[0188] in:

[0189] Each instance of G' is independently C1-C8 alkylene or C3-C8 cycloalkylene;

[0190] Each instance of L' is independently *-NR c C(=O)-; wherein * refers to the direction toward the pyrrolidine-2,5-dione ring;

[0191] R c Each instance of is independently H, C1-C 32 Alkyl or C2-C 32 alkenyl;

[0192] R is C4-C 32 Alkyl or C4-C 32 alkenyl; and

[0193] m is 0, 1 or 2;

[0194] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0195] In one embodiment, m is 0. In one embodiment, m is 1. In one embodiment, m is 2.

[0196] 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, the alkylene is C1 alkylene. In one embodiment, the alkylene is C2 alkylene. In one embodiment, the alkylene is C3 alkylene. In one embodiment, the alkylene is C4 alkylene. In one embodiment, the alkylene is C5 alkylene. In one embodiment, the alkylene is C6 alkylene. In one embodiment, the alkylene is -CH2-. In one embodiment, the alkylene is -CH2CH2-. In one embodiment, the alkylene is -CH2CH2CH2-.

[0197] In one embodiment, each instance of G' is independently C3-C8 cycloalkylene. In one embodiment, each instance of G' is independently 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.

[0198] In one embodiment, -(G'-L') m -R is In one embodiment, -(G'-L') m -R is

[0199] The descriptions provided herein for R apply to all formulae provided herein to the extent they are applicable.

[0200] In one embodiment, R is C4-C 32 In one embodiment, R is C6-C 32 In one embodiment, R is C8-C 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 C4 alkyl group. In one embodiment, R is a C6 alkyl group. In one embodiment, R is a C8 alkyl group. In one embodiment, R is a 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 alkyl group is a straight chain alkyl group. In one embodiment, the alkyl group is a branched chain alkyl group.

[0201] In one embodiment, R is C4-C 32 In one embodiment, R is C6-C32 In one embodiment, R is C8-C 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 C4 alkenyl. In one embodiment, R is C6 alkenyl. In one embodiment, R is C8 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 group is a straight chain alkenyl group. In one embodiment, the alkenyl group is a branched chain alkenyl group.

[0202] In one embodiment, R is unsubstituted. In one embodiment, R is -(CH2) 13-17 CH3.

[0203] In one embodiment, R is -L"-(CH2) 0-3 -R" where:

[0204] 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 -、*-NRa 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 * refers to the direction towards R;

[0205] R a Each instance of is independently H or C1-C6 alkyl;

[0206] R b Each instance of is independently H or C1-C6 alkyl;

[0207] x is 1 or 2; and

[0208] R" is a monocyclic, fused, bridged or spirocyclic 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.

[0209] In one embodiment, the ring portion is C3-C 10 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, the cycloalkyl is C3-C 12 Cycloalkyl. 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.

[0210] In one embodiment, the ring moiety is a 4- to 10-membered heterocyclyl containing 1 to 4 heteroatoms independently selected from O, N and S.

[0211] In one embodiment, R is substituted with -L"-R", wherein L" is absent, -O- or -C(=O)O-, and R" is a monocyclic, fused, bridged or spirocyclic alkyl group optionally substituted with one or more C1-C6 alkyl groups. 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 at the terminal position of R. In one embodiment, R is -(CH2) 14-18 -L"-R".

[0212] In one embodiment, R" is In one embodiment, R" is In one embodiment, R" is

[0213] In one embodiment, R is replace.

[0214] In one embodiment, R is substituted at the terminal position.

[0215] In one embodiment, the compound is a compound in Table 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof (wherein B is a modified or unmodified nucleobase).

[0216] Table 1 .

[0217]

[0218]

[0219]

[0220]

[0221]

[0222] For compounds with nucleobase B in Table 1 (e.g., Compound 1), it should be understood that the corresponding compounds wherein "B" is replaced by a specific nucleobase provided herein are also specifically provided herein. In one embodiment, the corresponding compounds wherein "B" is replaced by uracil (U) are particularly provided herein. In one embodiment, the corresponding compounds wherein "B" is replaced by cytosine (C) are particularly provided herein. In one embodiment, the corresponding compounds wherein "B" is replaced by adenine (A) are particularly provided herein. In one embodiment, the corresponding compounds wherein "B" is replaced by guanine (G) are particularly provided herein. In some embodiments, "Table 1" mentioned in the present application includes compounds modified with these nucleobases.

[0223] Oligonucleotides

[0224] In one embodiment, provided herein is an oligonucleotide comprising at least one lipophilic monomer of the formula:

[0225]

[0226] Where 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 (eg, as defined in formula (I), (III), (IV), respectively, including subformulae thereof).

[0227] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (V):

[0228]

[0229] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0230] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VA):

[0231]

[0232] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0233] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VA-1):

[0234]

[0235] or a pharmaceutically acceptable salt thereof.

[0236] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-A):

[0237]

[0238] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0239] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-A-1):

[0240]

[0241] or a pharmaceutically acceptable salt thereof.

[0242] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VB):

[0243]

[0244] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0245] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VB-1):

[0246]

[0247] or a pharmaceutically acceptable salt thereof.

[0248] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-B):

[0249]

[0250] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0251] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VI-B-1):

[0252]

[0253] or a pharmaceutically acceptable salt thereof.

[0254] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VII):

[0255]

[0256] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0257] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of (VII-A):

[0258]

[0259] or a pharmaceutically acceptable salt thereof.

[0260] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of (VII-A-1):

[0261]

[0262] or a pharmaceutically acceptable salt thereof.

[0263] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer of formula (VIII):

[0264]

[0265] or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0266] In one embodiment, the oligonucleotide comprises at least one lipophilic monomer in Table 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof (wherein B is a modified or unmodified nucleobase).

[0267] Table 2 .

[0268]

[0269]

[0270]

[0271]

[0272] For compounds with nucleobase B in Table 2 (e.g., compound 1'), it should be understood that the corresponding compounds in which "B" is replaced by a specific nucleobase provided herein are also specifically provided herein. In one embodiment, the corresponding compounds in which "B" is replaced by uracil (U) are particularly provided herein. In one embodiment, the corresponding compounds in which "B" is replaced by cytosine (C) are particularly provided herein. In one embodiment, the corresponding compounds in which "B" is replaced by adenine (A) are particularly provided herein. In one embodiment, the corresponding compounds in which "B" is replaced by guanine (G) are particularly provided herein. In some embodiments, "Table 2" mentioned in the present application includes compounds modified with these nucleobases.

[0273] In one embodiment, the oligonucleotide is an antisense, antagomir, microRNA, siRNA, pre-microRNA, antimir, ribozyme, RNA activator, U1 adaptor, immunostimulator or aptamer. In one embodiment, the oligonucleotide is a double-stranded RNA (dsRNA). In one embodiment, the oligonucleotide is a siRNA.

[0274] In one embodiment, the oligonucleotide is a siRNA comprising:

[0275] The antisense strand is complementary to the target gene;

[0276] A sense strand is complementary to the antisense strand.

[0277] In one embodiment, the lipophilic monomer is present in 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.

[0278] In one embodiment, the lipophilic monomer is conjugated to the terminal position (e.g., 3' end or 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 the terminal position (e.g., 3' end or 5' end) of the sense strand or 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.

[0279] In one embodiment, the lipophilic monomer is directly conjugated to the 3' end or the 5' end of the oligonucleotide. In one embodiment, the lipophilic monomer is conjugated to the 3' end or the 5' end of the oligonucleotide via a linking group.

[0280] In one embodiment, the lipophilic monomer of formula (V) (or any subformula thereof) is conjugated to the terminal position (e.g., 3' or 5' end) of the oligonucleotide. In one embodiment, the oligonucleotide comprises the following structure:

[0281]

[0282] in One or more nucleotides of an oligonucleotide comprising a 5'-3' direction. In one embodiment, A linker consisting of 1 to 3 nucleotides is included at the 3' end (ie, the O of the lipophilic monomer of formula (V) is directly linked to a nucleotide linker, which is linked to the remainder of the oligonucleotide). In one embodiment, the linker is dTdT. In one embodiment, Y is linked to hydrogen.

[0283] In one embodiment, the oligonucleotide comprises the structure:

[0284]

[0285] In one embodiment, the oligonucleotide comprises the structure:

[0286]

[0287] In one embodiment, the oligonucleotide comprises the structure:

[0288]

[0289] in One or more nucleotides of an oligonucleotide comprising a 5'-3' direction. In one embodiment, A linker consisting of 1 to 3 nucleotides is included at the 5' end (ie, Y of the lipophilic monomer of formula (V) is directly linked to a nucleotide linker, which is linked to the remainder of the oligonucleotide). In one embodiment, the linker is dTdT. In one embodiment, O is linked to hydrogen.

[0290] In one embodiment, the oligonucleotide comprises the structure:

[0291]

[0292] In one embodiment, the oligonucleotide comprises the structure:

[0293]

[0294] In one embodiment, the lipophilic monomer of formula (VII) (or any subformula thereof) is conjugated to the terminal position (e.g., 3' end or 5' end) of the oligonucleotide. In one embodiment, the oligonucleotide comprises the following structure:

[0295]

[0296] in One or more nucleotides of an oligonucleotide comprising in the 5'-3' direction.

[0297] In one embodiment, the oligonucleotide comprises the structure:

[0298]

[0299] In one embodiment, the oligonucleotide comprises the structure:

[0300]

[0301] in One or more nucleotides of an oligonucleotide comprising in the 5'-3' direction.

[0302] In one embodiment, the oligonucleotide comprises the structure:

[0303]

[0304] In one embodiment, the lipophilic monomer of formula (VIII) (or any subformula thereof) is conjugated to the terminal position (e.g., 3' end or 5' end) of the oligonucleotide. In one embodiment, the oligonucleotide comprises the following structure:

[0305]

[0306] in comprising one or more nucleotides of an oligonucleotide in the 5'-3' direction, and For the connector.

[0307] In one embodiment, the oligonucleotide comprises the structure:

[0308]

[0309] in comprising one or more nucleotides of an oligonucleotide in the 5'-3' direction, and For the connector.

[0310] In one embodiment, the linker is ("C6NH"). In one embodiment, the linker is ("C7NH"). The NH in a C6NH or C7NH linker is attached to a 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 C7NH linker is located at the 3' end of the oligonucleotide.

[0311] 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.

[0312] 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 following structure:

[0313]

[0314] Each of these One or more nucleotides independently comprising an oligonucleotide in the 5'-3' direction.

[0315] In one embodiment, the oligonucleotide comprises the structure:

[0316]

[0317] 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 following structure:

[0318]

[0319] Each of these One or more nucleotides independently comprising an oligonucleotide in the 5'-3' direction.

[0320] 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.

[0321] In one embodiment, the oligonucleotide further comprises a targeting ligand.

[0322] In one embodiment, there is provided herein a method for delivering an oligonucleotide to a cell, the method comprising contacting an oligonucleotide provided herein with a cell. In some embodiments, the cell is a brain cell. In some embodiments, the cell is located in a subject, such as a mouse, a non-human primate or a human. In one embodiment, compared with delivering the same oligonucleotide but not containing a lipid conjugate (lipophilic monomer) provided herein, the amount of the oligonucleotide delivered to the cell is increased by administration. In some embodiments, the amount of the increase of the oligonucleotide delivered is determined by the increase of the therapeutic activity of the oligonucleotide.

[0323] In one embodiment, the present invention provides a method for reducing the expression of a target gene in a cell, the method comprising contacting the cell with an oligonucleotide provided herein. In one embodiment, the present invention provides a method for reducing the expression of a target gene in a subject, the method comprising administering an oligonucleotide provided herein to the subject.

[0324] In one embodiment, without being limited to a particular theory, the oligonucleotides provided herein comprising at least one lipophilic monomer exhibit improved pharmacodynamic characteristics compared to corresponding oligonucleotides that do not comprise a lipophilic monomer.

[0325] Chemical modification of nucleotides

[0326] In specific embodiments, the dsRNA (e.g., siRNA) agents described herein include one or more nucleotide modifications. Nucleotide modifications include, for example, terminal modifications, such as 5' terminal modifications (e.g., phosphorylation, conjugation, reverse connection) or 3' terminal modifications (e.g., conjugation, DNA nucleotides, reverse connection, etc.); base modifications, such as replacement of bases with stable bases, destabilizing bases, or bases paired with amplified partner pools, removal of bases (absaccharide nucleotides), or conjugation of bases; sugar modifications (e.g., at the 2' or 4' position) or sugar replacement; or backbone modifications, including modification or replacement of phosphodiester bonds. In some embodiments, the dsRNA agent includes at least one modification selected from the group consisting of modified internucleoside bonds, modified nucleobases, modified sugars, and any combination thereof. Without limitation, such modifications may be present at any position in the dsRNA agent (e.g., in the sense strand, antisense strand, or both strands).

[0327] In some embodiments, the dsRNA agent comprises one or more modified sugar modifications, such as one or more substituted sugar moieties. In some embodiments, the dsRNA agents described herein include one of the following at the 2' position: H; F; or OCH3(OMe).

[0328] In some embodiments, the dsRNA agents described herein include one or more glycol nucleic acids (GNAs). Typically, GNAs are acyclic nucleic acid analogs in which the repeating glycol units are linked by phosphodiester bonds, unlike the ribose-phosphodiester backbone of RNA.

[0329] In certain embodiments, the dsRNA agents described herein include one or more terminal modifications, such as 5' terminal phosphorylation, conjugation, or reverse ligation. In some embodiments, the terminal modification includes a 5'-phosphate, such as a 5' terminal phosphate on the antisense strand of the dsRNA agent.

[0330] In some embodiments, the dsRNA agent includes a sense strand and / or an antisense strand having reverse abasic nucleotides. In one embodiment, the sense strand contains reverse abasic nucleotides at the 3' end. In another embodiment, the sense strand contains reverse abasic nucleotides at the 5' end. In some embodiments, the sense strand contains reverse abasic nucleotides at the 5' and 3' ends.

[0331] In some embodiments, the dsRNA agent comprises a phosphate or a phosphate mimetic at the 5' end of the antisense strand. In one embodiment, the phosphate mimetic is 5'-vinylphosphonate (VP).

[0332] In some embodiments, the dsRNA agent comprises one or more modified nucleotides. In specific 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, reverse abasic nucleotides, nucleotides comprising S-glycol nucleic acid (GNA), unlocked nucleic acids, 5'-vinylphosphonate-2'-O-methyl-uridine and combinations thereof.

[0333] In some embodiments, the dsRNA agent comprises one or more modified internucleoside bonds (i.e., modified RNA backbones). The modified internucleoside bonds include, for example, phosphorothioates (e.g., monothiophosphates). 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, the cation of the salt (e.g., sodium cation) is present in the agent in the form of counterions of substantially all electronegative groups (e.g., phosphodiester and / or phosphorothioate groups) present in the agent. In some embodiments, the counterion is a condensed counterion. In a specific embodiment, the counterion is a condensed sodium cation. In some embodiments, the condensed counterion is hydrated. In a specific embodiment, the condensed counterion is a hydrated sodium cation. A reagent in which substantially all phosphodiester and / or phosphorothioate bonds have counterions includes no more than 5, 4, 3, 2, or 1 phosphodiester and / or phosphorothioate bonds without counterions. In other words, the electronegativity of the dsRNA reagent is neutralized or substantially neutralized by the condensation of counterions surrounding the dsRNA. In some embodiments, when the dsRNA reagent described herein is in the form of a sodium salt, sodium ions are present around the reagent in the form of counterions for substantially all phosphodiester and / or phosphorothioate groups present in the reagent.

[0334] The phosphate group of the phosphodiester bond between nucleosides can be modified by replacing one of the oxygens with different substituents. One result of this modification can increase the resistance of the oligonucleotide to nucleic acid degradation. Another result of this modification can improve the stability of the single-stranded RNA (ssRNA) hybridized in the dsRNA reagent. The example of the modified phosphate group includes thiophosphate (e.g., monothiophosphate).

[0335] In some embodiments, the dsRNA agent comprises an RNA mimetic in which both the sugar and the internucleoside linkage (ie, the backbone) of the nucleotide unit are replaced by alternative groups. In specific embodiments, the base unit is maintained to hybridize with an appropriate target sequence.

[0336] The dsRNA agents described herein may contain one or more asymmetric centers, thus giving rise to enantiomers, diastereomers, and other stereoisomeric configurations, which may be defined by absolute stereochemistry as (R) or (S), such as for sugar anomers, or as (D) or (L), such as for amino acids. The dsRNA agents provided herein include all such possible isomers, as well as their racemic and optically pure forms.

[0337] The part connected to the nucleotide sequence

[0338] In some embodiments, dsRNA agents as described herein are conjugated with one or more non-nucleotide groups.Non-nucleotide groups can, for example, enhance the targeting, delivery or attachment of dsRNA agents.Non-nucleotide groups can be covalently connected with 3' ends, 5' ends and / or the inside of the sense strand or antisense strand of dsRNA agents.Non-nucleotide groups can be covalently connected with 3' ends, 5' ends, 3' ends and 5' ends, inside, 3' ends and inside, 5' ends and inside of the sense strand and / or antisense strand of dsRNA agents, or at 3' ends, 5' ends and inside.In some embodiments, dsRNA agents as described herein contain non-nucleotide groups covalently connected with 3' ends, 5' ends, 3' ends and 5' ends, inside, 3' ends and inside, 5' ends and inside of the sense strand.In certain embodiments, dsRNA agents as described herein contain non-nucleotide groups connected with the 5' ends of the sense strand. In certain embodiments, the dsRNA agents described herein contain a non-nucleotide group connected to the 3' end of the sense strand. In certain embodiments, the dsRNA agents described herein contain a non-nucleotide group connected to the interior of the sense strand. The non-nucleotide group can be directly or indirectly connected to the dsRNA agent through a linker / connecting group.

[0339] In some embodiments, the linking group is conjugated to the dsRNA agent. The linking group facilitates the covalent connection of the agent to the targeting ligand or the delivery polymer or the delivery vehicle. The linking group can be connected to the 3' end, 5' end and / or the interior of the sense strand of the dsRNA agent. In some embodiments, the linking group is connected to the sense strand of the dsRNA agent. In some embodiments, the linking group is conjugated to the 5' end, 3' end and / or the interior of 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 of the dsRNA agent. In some embodiments, the linking group is conjugated to the interior of the sense strand of the dsRNA agent.

[0340] Typically, a linker or linking group is a connection between two atoms that connects a chemical group (such as a dsRNA agent) or a segment of interest to another chemical group (such as a targeting group or a delivery polymer) or a segment of interest through one or more covalent bonds. Unstable connections contain unstable bonds. The connection may optionally include a spacer that increases the distance between the two joined atoms. The spacer may further increase the flexibility and / or length of the connection.

[0341] Any dsRNA agent nucleotide sequence listed in Table 3 (whether modified or unmodified) can contain a 3'-end, 5'-end and / or internal targeting ligand and / or linking group. Any dsRNA agent duplex listed in Table 1, Table 2, Table 3, Table 4, Table 5 or Table 9 (whether modified or unmodified) can also contain a targeting ligand and / or linking group, and the targeting ligand or linking group can be attached to the 3'-end, 5'-end and / or the interior of the sense strand or antisense strand of the dsRNA agent duplex.

[0342] Delivery vehicle

[0343] In some embodiments, a dsRNA agent described herein can be delivered to a cell or tissue using a delivery vehicle.

[0344] Composition

[0345] In one aspect, provided herein are compositions (eg, pharmaceutical compositions) comprising a dsRNA agent described herein. The composition may also comprise a pharmaceutically acceptable carrier.

[0346] It should be understood that the above detailed description and the attached examples are illustrative only and should not be considered as limiting the scope of the subject matter. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Without departing from the spirit and scope of the present invention, such changes and modifications may be made, including but not limited to those changes and modifications related to the chemical structures, substituents, derivatives, intermediates, synthesized, formulations and / or methods of use provided herein. The United States Patents and publications cited herein are incorporated herein by reference.

[0347] Example

[0348] Certain embodiments of the invention are illustrated by the following non-limiting examples.

[0349] Experimental procedures for the synthesis of oligonucleotide lipid conjugates

[0350] Synthesis was performed by the phosphoramidite method on solid phase following standard RNA synthesis protocols. All oligonucleotides were prepared on a MerMade 12 synthesizer at a 10 μmole scale using generic or custom vectors. A typical synthesis cycle included the following four steps: 1. detritylation; 2. coupling; 3. capping; and 4. oxidation / sulfurization.

[0351] Monomers of RNA phosphoramidites with standard protecting groups 5'-O-dimethoxytrityl-N 6 -benzoyl-2'-O-(tert-butyldimethylsilyl)-adenosine 3'-O-(N,N'-diisopropyl-cyanoethylphosphoramidite), 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-cyanoethylphosphoramidite) and 5'-O-dimethoxytrityl-2'-O-(tert-butyldimethylsilyl)uridine 3'-O-(N,N'-diisopropyl-2-cyanoethylphosphoramidite) were used for the synthesis.

[0352] The 2'-modified monomers consisted of 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-cyanoethylphosphoramidite) and 5'-O-dimethoxytrityl-N 2-isobutyryl-2'-OMe-guanosine 3'-O-(N,N'-diisopropyl-2-cyanoethylphosphoramidite).

[0353] All phosphoramidites were used at a concentration of 0.1 M in acetonitrile, except guanosine, which was used at a concentration of 0.1 M in 20% DMF / MeCN. Detritylation was performed for 3 minutes with a 3% dichloroacetic acid solution in dichloromethane. Coupling was performed with a 0.45M solution of 5-(ethylthio)tetrazolium in MeCN, and the coupling time was 12 minutes. Internucleotide phosphites were oxidized to phosphates using a standard 0.02M iodine solution of MeCN / pyridine / water (70:20:10). Phosphothioates were introduced by oxidizing phosphites to thiophosphates using a 0.1M solution of hydrogenated xanthin in pyridine. End-capping was performed with THF / acetic anhydride / pyridine (80:10:10) and 1-methylimidazole / acetonitrile (20:80 v / v) solutions. The complete synthesis cycle was 30 minutes.

[0354] Lipid bioconjugation

[0355] Lipids are incorporated into the sense strand by on-column and / or post-column synthesis. For on-column synthesis, lipids at the terminal (3' and 5') and internal positions are synthesized on a solid support. For post-column synthesis, lipids are introduced into the solution phase along with their corresponding N-hydroxysuccinimide esters and the additional amine linker of the nucleotide.

[0356] On-column synthesis

[0357] For example, the on-column introduction of compound 17 is performed at the 3' end of the sense strand using a 3'-amino modifier (such as 2-dimethoxytrityloxymethyl-6-fluorenylmethoxycarbonylamino-hexane-1-succinyl)-long chain alkylamino-CPG S1-1) (Scheme 1). After deprotection of the Fmoc group with a 20% piperidine solution in DMF, the CPG is washed thoroughly with DMF, MeCN, and ether and dried. A solution of 80 μmol of compound 17 in 2 ml of 1,4-dioxane is added to the above CPG, followed by addition of 30 μl of DIPEA and shaking for 12 hours. The CPG is washed with DCM, MeCN, and Et2O. 1 ml of a solution of CAP A and CAP B is added and shaken for 30 minutes. S1-2 is washed with DCM, MeCN, Et2O, dried, and used for oligomer assembly.

[0358] Solution 1

[0359]

[0360] Compound 17 was introduced at the 5' end of the sense strand using a 5'-amino modifier such as 6-(4-monomethoxytritylamino)hexyl-(2-cyanoethyl)-(N,N'-diisopropyl)-phosphoramidite. After assembling the sense strand on CPG according to standard procedures, in the penultimate step, the 5' end of the sense strand was coupled with 6-(4-monomethoxytritylamino)hexyl-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite, followed by deblocking of the 4-monomethoxytrityl group with 3% DCA / DCM for 30 minutes. CPG was subsequently washed with DCM, ACN and coupled with compound 17 for 12 hours (Scheme 2).

[0361] Solution 2

[0362]

[0363] During the sense strand assembly process, an adamantyl lipid is introduced at the internal position together with the corresponding phosphoramidite monomer. For example, Scheme 3 illustrates this with the uridine-based phosphoramidite monomer 25.

[0364] Solution 3

[0365]

[0366] Lysis, purification and desalting

[0367] After the synthesis is completed, the oligonucleotide is cracked from the carrier, and ammonia solution is used at 55 ℃ by core base and phosphate group deprotection 7 hours.CPG is filtered and washed with ethanol / acetonitrile / water (3:1:1v / v).After reducing the volume, oligonucleotide is purified by reverse phase or ion exchange chromatography.The buffer for reverse phase is 0.1M sodium acetate (buffer A) and acetonitrile (buffer B) in 90% / 10% water / acetonitrile, and the buffer for ion exchange is 20mM sodium phosphate (pH 11) (buffer A) in 90% / 10% water / acetonitrile and 20mM sodium phosphate (pH 11), 1.8M sodium bromide (buffer B) in 90% / 10% water / acetonitrile.The fraction containing full-length oligonucleotide is merged, desalted, and analyzed by liquid chromatography-mass spectrometry (LC-MS) compound.

[0368] Post-column synthesis

[0369] After purification and desalting of the corresponding oligonucleotides, lipids were bioconjugated at the terminal positions (3', 5' ends) and internal positions of the sense strand in solution phase.

[0370] For example, incorporation at internal positions is performed using phosphoramidite monomers derivatized with an amine linker at the 2' position of the corresponding nucleotide. Scheme 4 illustrates this using uridine-based phosphoramidite monomer S4-1. After incorporation of S4-1 at internal positions and completion of the assembly, cleavage, purification, and desalting processes, free oligonucleotides with side chain amines are coupled to the corresponding lipid-NHS esters in solution phase.

[0371] Solution 4

[0372]

[0373] Bioconjugation at the terminal / internal position was performed with 0.15 mM deprotected and desalted oligonucleotide solution in 0.1 M NaHCO3 solution (pH 8.4) and 1.5 mM corresponding lipid-NHS ester solution in DMF at 60°C. A DMF solution (0.6 mL) of lipid-NHS ester was added to the oligonucleotide solution (0.25 mL), 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 was typically >90% complete in less than one hour. The reaction mixture was diluted with water and purified by RP-HPLC (C-8Xbridge Waters column; A: 50 mM NaOAc, B: MeCN; 5%-100% B gradient, 60°C). The isolated yield of lipid-conjugated oligonucleotides was 60%-70%.

[0374] Duplex formation

[0375] To form duplexes, equimolar amounts of sense and antisense strands were mixed and kept at room temperature for 30 minutes. The integrity of the duplexes was confirmed by denaturing and native HPLC analysis.

[0376] Example 1 .

[0377]

[0378] Preparation of 1-2

[0379] At 0 ° C, under an inert atmosphere of nitrogen, TIPDSCl2 (111.0 g, 354.0 mmol) was added to a suspension of 1-1 (72.0 g, 295.0 mmol) in DMF (720 mL). Imidazole (50.1 g, 737.5 mmol) was then added, and the resulting mixture was stirred at room temperature for 14 hours. The mixture was diluted with EtOAc (2.5 L), washed with H2O, saturated aqueous sodium bicarbonate solution and brine, and dried over Na2SO4. The evaporated residue was purified by column chromatography (SiO2, 0-10% MeOH / DCM) to give 1-2 (115.0 g, 236 mmol, 80% yield) as a white solid. MS: m / z 487 [M+H] + .

[0380] Preparation of 1-3

[0381] 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 two-phase solution (1.0 / 2.0 L) of DCM / H2O, and then BzCl (16 mL, 138.7 mmol) was added to the mixture under 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 phase was collected, and the aqueous phase was extracted with DCM. The combined organic extracts were dried over Na2SO4 and concentrated. The evaporated residue was purified by silica gel column chromatography (PE / EtOAc 3 / 1). 1-3 (29.0 g, 53.0% yield) was obtained as a white solid. MS: m / z 591 [M+H] + . 1 H NMR (DMSO-d6): δ8.00 (d, J=7.4 Hz, 2H), 7.86 (d, J=8.0 Hz, 1H), 7.79 (t, J=7.4 Hz, 1H), 7.60 (t, J=7.8 Hz, 2H), 5.82 (d, J=8.0 Hz, 1H), 5.66 (d, J=4.4 Hz, 1H, exchanged 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).

[0382] Preparation of 1-4

[0383] To a solution of 1-3 (10.5 g, 17.8 mmol), methyl methacrylate (30.5 g, 356 mmol) in t-BuOH (50 mL) was added Cs2CO3 (2.9 g, 8.9 mmol). The reaction mixture was stirred at 30 ° C for 4 hours and then diluted with EtOAc. The mixture was washed twice with H2O, dried over Na2SO4 and concentrated. The evaporated residue was purified by column chromatography (SiO2, PE / EtOAc 6 / 1) to give 1-4 (8.7 g, 72% yield) as a white solid. MS: m / z 677 [M+H] + .

[0384] Preparation of 1-5

[0385] 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 hours. It was then diluted with EtOAc / water (1 / 1), the organic layer was washed with water, dried over Na2SO4 and concentrated. The residue was purified by flash preparative HPLC (C 18 Purification was performed by column, mobile phase CH3CN and H2O (0.5% NH4HCO3, gradient 50% to 100% MeCN) to give 1-5 (6.1 g, 82%) as a white solid. MS: m / z 573 [M+H] + .

[0386] Preparation of 1-6

[0387] 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 hour, then concentrated, and the residue was purified by column chromatography (SiO2, 0-5% MeOH / EtOAc). This gave 1-6 (3.2 g, 90% yield) as a white solid. MS: m / z 331 [M+H] + ; 1 H NMR (DMSO-d6): δ11.32 (s, 1H, exchanged with D2O), 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 D2O), 5.03 (d, J=5.0 Hz, 1H, exchanged with D2O), 4.10 (m, 1H), 3.92 (m, 1H), 3.84-3.70 (m, 3H), 3.65-3.53 (m, 3H), 2.56 (m, 2H).

[0388] Preparation of 1-7

[0389] To a solution of 1-6 (3.2 g, 9.6 mmol) in THF / H2O (30 / 15 mL) was added KOH (1.6 g, 28.8 mmol). The reaction mixture was stirred at 20°C for 1 hour. The solution was washed with H + The exchange resin was neutralized to pH 7, filtered and concentrated to give 1-7 as a white solid. MS: m / z 317 [M+H] + .

[0390] Preparation of 1-8

[0391] To a solution of 1-7 (3.0 g, 9.6 mmol) in DMF (30 mL) was added EDCI (1.8 g, 9.6 mmol), HOBT (1.3 g, 9.6 mmol), hexadecane-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 hours and then diluted with EtOAc. The mixture was washed with H2O and 5% citric acid, dried over Na2SO4 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-d6): δ11.32 (s, 1H, exchanged with D2O), 7.91 (d, J=8.0 Hz, 1H), 7.88 (t, J=5.2 Hz, 1H, exchanged with D2O), 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 D2O), 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).

[0392] Preparation of 1-9

[0393] Under N2 atmosphere, DMTrCl (2.4 g, 7.1 mmoll) was added to a solution of 1-8 (3.2 g, 5.9 mmol) in pyridine (30 mL). The reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was then diluted with EtOAc, washed with H2O 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 [MH] - ; 1 H-NMR (DMSO-d6): δ11.38 (s, 1H, exchanged with D2O), 7.87 (t, J = 5.2 Hz, 1H, exchanged with D2O), 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,exchanged with D2O),4.24(m,1H),4.03-3.92(m,2H),3.79(m,2H),3.74(s,6H),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).

[0394] Preparation of 1

[0395] Under N2 atmosphere, DCI (418 mg, 2.9 mmol) and CN(CH2)2OP[N(iPr)2]2 (1.2 g, 4.1 mmol) were added to a solution of 1-9 (2.9 g, 3.4 mmol) in DCM (30 mL) under N2 atmosphere. The reaction mixture was stirred at 30°C for 2 hours. The solution was then diluted with DCM and washed with H2O. The organic layer was washed with brine, dried over Na2SO4 and concentrated. The residue was purified by flash preparative HPLC (C 18 Column; mobile phase H2O with 0.5% NH4HCO3, CH3CN; gradient 0-100% MeCN) to obtain compound 1 (2.6 g, 71% yield) 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.

[0396] Example 2 .

[0397]

[0398] Preparation of 2-1

[0399] 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] + .

[0400] Preparation of 2-2

[0401] To a solution of 2-1 (11.5 g, 19.9 mmol) in anhydrous DCM (100 mL) was added a solution of hexadecane-1-amine (4.8 g, 19.9 mmol) in anhydrous DCM (100 mL). The reaction mixture was stirred at 30 ° C for 16 hours, then quenched with H2O and diluted with DCM. The organic phase was washed with H2O and 5% citric acid and dried over Na2SO4. The evaporated residue was purified by silica gel column chromatography (0-20% acetone / EtOAc / DCM 1 / 1) to give 2-2 (11.1 g, 70% yield) 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=8.0 Hz, 1H), 7.33 (t, J=5.5 Hz, 1H, exchanged with D2O), 5.66 (d, J=1.6 Hz, 1H), 5.59 (d, J=8.0 Hz, 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).

[0402] Preparation of 2-3

[0403] 1M TBAF in THF (7.4 mL, 7.4 mmol) was added dropwise to a solution of 2-2 (4.7 g, 6.2 mmol) in THF (50 mL). The reaction mixture was stirred at room temperature for 1 hour. EtOAc was then added, and the mixture was washed with H2O and dried over Na2SO4. 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-d6): δ11.33 (s, 1H, exchanged with D2O), 7.90 (d, J=8.0 Hz, 1H), 7.27 (t, J=5.5 Hz, 1H, exchanged with D2O), 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 D2O), 5.19 (m, 1H, exchanged with D2O), 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).

[0404] Preparation of 2-4

[0405] Under N2 atmosphere, DMTrCl (2.2 g, 6.6 mmoll) was added to a solution of 2-3 (3.1 g, 6.0 mmol) in pyridine (30 mL). The reaction mixture was stirred at room temperature for 3 hours, then diluted with EtOAc, washed with H2O and dried over Na2SO4. The evaporated 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 812 [MH] - ; 1 H-NMR (DMSO-d6): δ11.39 (s, 1H, exchanged with D2O), 7.71 (d, J=8.0 Hz, 1H), 7.39-7.21 (m, 10H), 6.91-6.89 (m, 4H), 5.91 (d, J=4.8 Hz, 1H), 5.52 (d, J=5.6 Hz, 1H, exchanged with D2O), 5.37 (d, J=8.0 Hz, 1H), 5.14 (t, J=5.1 Hz, 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).

[0406] 2. Preparation

[0407] Under N2 atmosphere, DCI (234 mg, 1.9 mmol) and CN(CH2)2OP[N(iPr)2] (843 mg, 2.8 mmol) were added to a solution of 2-4 (1.9 g, 2.3 mmol) in DCM (20 mL) under N2 atmosphere. The reaction mixture was stirred at 30 °C for 2 hours. The solution was then diluted with DCM and washed with H2O. The organic layer was washed with brine, dried over Na2SO4 and evaporated. The residue was purified by silica gel column chromatography (PE / EtOAc 2 / 1 to 1 / 2) to give compound 2 (2.1 g, 86% yield) as a white solid. MS: m / z 1012 [MH] - ; 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.

[0408] Example 3 .

[0409]

[0410] Preparation of 3-3

[0411] 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 hours and then concentrated. The residue was distributed between dichloromethane and water. The organic layer was washed with sodium bicarbonate aqueous solution and brine and dried over MgSO4. The evaporated residue was co-evaporated with toluene to obtain compound 3-2 as a viscous oil. CCl3CN (140.0 g) and TEA (3 mL) were added, and the solution was heated at 90 ° C for 16 hours. 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 obtain compound 3-3 (89.1 g, 97% yield) in a foamy state. MS: m / z 670 [MH] - .

[0412] Preparation of 3-4

[0413] 6N NaOH solution (180 mL) was added to a solution of 3-3 (89.1 g, 132.8 mmol) in ethanol (360 mL), and the reactant was refluxed at 70 ° C for 16 hours, then cooled and concentrated. The residue was distributed between dichloromethane and saturated ammonium chloride. The aqueous phase was washed with dichloromethane, and the combined organic extracts were dried over magnesium sulfate, filtered and evaporated. The residue was purified by silica gel column chromatography (DCM / MeOH 30 / 1 to 10 / 1 containing 1% TEA) to give compound 3-4 (42.0 g, 77.1 mmol, 56.2% yield over 3 steps) as a white foam. MS: 544m / z[MH] - . 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).

[0414] Preparation of 3-5

[0415] 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) were added to a solution of 3-4 (4.2 g, 7.7 mmol) in anhydrous DMF (42 mL). The reaction mixture was stirred at room temperature for 15 hours, 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 [MH] - . 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).

[0416] 3. Preparation

[0417] Under N2 atmosphere, CN(CH2)2OP[N(iPr)2](1.3g, 4.4mmol) and DCI(341mg, 2.9mmol) were added to a suspension of 3-5(2.7g, 3.4mmol) in anhydrous DCM(30mL). The mixture was stirred at room temperature for 2.5 hours. The solution was then washed with water, brine and dried over Na2SO4. The concentrated residue was purified by silica gel column chromatography (PE / EtOAc 5 / 1 to 1 / 3) to give compound 3(2.4g, 2.4mmol, 70% yield) as a white solid. MS:m / z 982[MH] - . 1 H 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.

[0418] Example 4 .

[0419]

[0420] Preparation of 4-1

[0421] A solution of 3-4 (2.1 g, 3.9 mmol) and hexadecyl isocyanate (1.1 g, 3.9 mmol) in DMF (20 mL) was stirred at 30 ° C for 2 hours. 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] + . 1 H-NMR (DMSO-d6): δ11.30 (s, 1H, exchanged with D2O), 7.63 (d, J = 8 Hz, 1H), 7.41-7.21 (m, 9H), 6.90 (d, J = 8 Hz, 4H), 6.27 (t, J = 5 Hz, 1H, exchanged with D2O), 5.98 (d, J = 8 Hz, 1H, exchanged with D2O), 5.86 (d, J = 5 Hz, 1H), 5.78 (d, J = 8 Hz, 1H, exchanged with D2O), 5.41-5.39 (m, 1H), 4.45 (dd, J = 8 Hz, 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).

[0422] 4. Preparation

[0423] Under N2 atmosphere, DCI (313 mg, 2.2 mmol) and CN(CH2)2OP[N(iPr)2] (932 mg, 3.1 mmol) were added to a solution of 4-1 (2.1 g, 2.6 mmol) in DCM (20 mL) under N2 atmosphere. The mixture was stirred at 30 °C for 2 hours, then diluted with DCM, washed with H2O and dried over Na2SO4. The evaporated 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,4H),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,1H),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.

[0424] Example 5 .

[0425]

[0426] Preparation of 5-2

[0427] 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 allowed to warm 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 another 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), 2N hydrochloric acid (2×200 mL), saturated aqueous NaHCO3 (2×400 mL), water (2×500 mL) and saturated brine (2×400 mL). The organic layer was dried (anhydrous Na2SO4), filtered and concentrated under reduced pressure. The crude product was purified on a silica gel column using petroleum ether / ethyl acetate (50:1-20:1) to obtain 160 g (50%) of 5-2 as a yellow liquid. MS: m / z 499.30 [M+Na] + .

[0428] Preparation of 5-3

[0429] At 0 ° C, under an argon atmosphere, a solution of 5-2 (160 g, 336 mmol) and p-toluene thiophenol (50 g, 403 mmol, 1.2 equivalents) in anhydrous dichloromethane (1600 mL) was added dropwise with a solution of tin tetrachloride in dichloromethane (1M, 74 mL). The resulting solution was stirred at room temperature for 3 hours. The reactants were quenched with trimethylamine (50 mL) and concentrated. The crude residue was purified on a silica gel column with petroleum ether / ethyl acetate (100: 1-80: 1) to give 100 g (55%) of 5-3 as a light yellow oil. MS: m / z 563.20 [M+Na] + . 1 H 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).

[0430] Preparation of 5-4

[0431] At -20 ° C, under Ar, 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) was added N-iodosuccinimide (36.7 g, 162.9 mmol, 1.1 equiv) and silver trifluoromethanesulfonate (3.8 g, 14.8 mmol, 0.1 equiv). The resulting solution was stirred at -20 ° C for 30 minutes, quenched with trimethylamine (30 mL) and concentrated. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (50: 1-20: 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).

[0432] Preparation of 5-5

[0433] At 0 ° C, under Ar atmosphere, to a solution of 5-4 (34 g, 65.8 mmol) in methanol (340 mL) was added a solution of NaOMe in MeOH (5 M, 19.7 mL). The resulting solution was stirred at 0 ° C for 1 hour, then quenched with glacial acetic acid and concentrated. The crude product was loaded onto a silica gel column and eluted with petroleum ether / ethyl acetate (40: 1-10: 1) to give 24.5 g (78% yield) of 5-5 as a yellow oil. MS: m / z 498.25 [M+Na] + .

[0434] Preparation of 5-6

[0435] Under Ar, acrylonitrile (56.4 g, 1052.6 mmol, 20 equivalents) and cesium carbonate (17.2 g, 52.6 mmoL, 1.0 equivalents) were added to a solution of 5-5 (25 g, 52.6 mmol) in tert-butyl alcohol (250 mL) and stirred at room temperature for 2 hours. The resulting mixture was filtered and the filtrate was concentrated. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (30: 1-8: 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).

[0436] Preparation of 5-7

[0437] To 5-6 (10 g, 18.9 mmol) in 100 mL 7M NH3 in methanol was added Raney nickel (50%, 5 g). The mixture was hydrogenated at 50 ° C under 5 atm hydrogen pressure for 24 hours. 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. It was used in the next step without further purification. MS m / z[M+H] + (ESI):507.30.

[0438] Preparation of 5-8

[0439] At 0 ° C, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (5.3g, 27.6mmol, 2.5 equivalents), N, N- diisopropylethylamine (7.2g, 55.3mmoL, 5.0 equivalents) and 1-hydroxybenzotriazole (3.73g, 27.65mmol, 2.5 equivalents) were added to a solution of 5-7 (5.6g, 11.1mmol) and palmitic acid (6.2g, 24.4mmol, 2.2 equivalents) in dichloromethane (56mL) under Ar. The reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was diluted with dichloromethane (400mL) 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: 1-80: 1) to obtain 6.0g (55%) of 5-8 as a white solid. MS: m / z 983.90[M+H] + . 1 HNMR(CDCl3): δ6.24(d,J=24.1Hz,2H),4.83(s,1H),4.45(dd,J=8.3,4.3Hz,1H),3.9 8-3.94(m,3H),3.92-3.85(m,1H),3.78-3.69(m,2H),3.67(d,J=4.3Hz,1H),3.54-3. 41(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).

[0440] Preparation of 5-9

[0441] Under Ar, triethylamine (3.69g, 36.6mmol) was added to a solution of triethylamine trihydrofluoride (2.95g, 18.3mmol) in anhydrous THF (60mL) and stirred at room temperature for 10 minutes. A solution of 5-8 (6g, 6.1mmol) in anhydrous THF (60mL) was added and the reaction mixture was stirred at room temperature for 1 hour. It was then diluted with dichloromethane (300mL), washed with water and saturated brine. The organic layer was dried (Na2SO4), filtered and concentrated. The residue was dissolved with 10mL dichloromethane and slowly poured into 200mL acetonitrile under stirring. Solid was collected by filtration. 5-9 of 3g (66% yield) as a white solid was obtained. MS: m / z 742.00[M+H] + .

[0442] Preparation of 5-10

[0443] At 0 ° C, 4,4'-dimethoxytrityl chloride (1.54 g, 1.53 mmoL, 1.2 equivalents) was added to a solution of 5-9 (2.8 g, 3.78 mmol) in pyridine (60 mL) under Ar and stirred at 25 ° C for 4 hours. The reaction mixture was quenched with methanol (10 mL), diluted with dichloromethane (200 mL), washed with saturated sodium bicarbonate aqueous solution and saturated brine. The organic layer was dried (Na2SO4), filtered and concentrated. The residue was purified on a silica gel column with dichloromethane / methanol (100: 1-60: 1) to give 3.1 g (79% yield) of 5-10 as a light yellow oil. MS: m / z 1041.75 [MH] - . 1 H NMR(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).

[0444] 5. Preparation

[0445] Under Ar, 3-(bis(diisopropylamino)phosphinoyl)propionitrile (1.1 g, 3.6 mmoL, 1.2 equiv.), 4,5-dicyanoimidazole (386 mg, 3.3 mmol, 1.1 equiv.) were added to a solution of 5-10 (3.1 g, 3 mmol) in dichloromethane (31 mL), and stirred at room temperature for 40 minutes. The resulting solution was diluted with dichloromethane (500 mL), washed with saturated aqueous sodium bicarbonate solution and saturated brine. The organic layer was dried (Na2SO4), filtered and concentrated. The crude product was purified by Prep-Flash (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] + . 1 H NMR(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 P NMR(DMSO-d6): δ148.86,148.57.

[0446] Example 6 .

[0447]

[0448] Preparation of 6-2

[0449] At -20 ° C, NIS (23.3 g, 103.6 mmol, 1.1 equiv) and silver trifluoromethanesulfonate (2.4 g, 9.4 mmol, 0.1 equiv) were added 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) under Ar. The resulting solution was stirred at -20 ° C for 1 hour, 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: 1-10: 1) to give 25 g (58% yield) of 6-2 as a yellow oil. MS: m / z 473.30 [M+NH4] + .

[0450] Preparation of 6-3

[0451] At 0 ° C, under Ar, to a solution of 6-2 (25 g, 54.9 mmol) in MeOH (250 mL) was added a methanol solution of NaOMe (5 M, 13.2 mL). The resulting mixture was stirred at 0 ° C for 0.5 hours, 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 6-3 as a yellow oil. MS: m / z 431.30 [M+Na] + .

[0452] Preparation of 6-4

[0453] At 0 ° C, sodium hydride (60% dispersion in oil, 9.68 g, 242 mmol, 5.0 equivalents) was added to a solution of 6-3 (20 g, 48.42 mmol) in DMF (200 mL) and stirred at 0 ° C for 30 minutes. 1-Bromo-2-methoxyethane (13.3 g, 96.9 mmol, 2.0 equivalents) was added and the resulting solution was stirred at room temperature for 16 hours. 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 Na2SO4), filtered and concentrated. The crude product was applied to a silica gel column and eluted with dichloromethane / methanol (100: 1-80: 1) to give 16 g (70% yield) of 6-4 as a yellow oil. MS: m / z 489.30 [M+Na] + . 1H NMR(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).

[0454] Preparation of 6-5

[0455] Triphenylphosphine (17.8 g, 67.9 mmol, 2.0 equiv.) was added to a solution of 6-4 (16 g, 34 mmol) in a mixture of 320 mL of tetrahydrofuran and water (10: 1). The reaction mixture was stirred at room temperature for 12 hours and then concentrated. The residue was purified by flash preparative HPLC (C18 column; mobile phase, 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] + .

[0456] Preparation of 6-6

[0457] 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 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. The reaction mixture was stirred at room temperature for 12 hours, then diluted with dichloromethane (200 mL) and washed with water. The organic layer was dried (anhydrous Na2SO4), filtered and concentrated under reduced pressure. The residue was purified on a silica gel column with dichloromethane / methanol (100: 1-60: 1) to give 11 g (55% yield) of 6-6 as a yellow oil. MS m / z706.45[M+Na] + .

[0458] Preparation of 6-7

[0459] Pd / C (10%, 1.1 g) and acetic acid (5 mL) were added to a solution of 6-6 (11 g, 16.10 mmol) in MeOH / THF (1: 1, 110 mL) and stirred for 8 hours under H2 atmosphere. The mixture was filtered and the filtrate was concentrated. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (20: 1-4: 1) to give 7.5 g (92% yield) of 6-7 as a white solid. MS: m / z): 526.40 [M+Na] + .

[0460] Preparation of 6-8

[0461] At 0 ° C, 4,4'-dimethoxytrityl chloride (6.04 g, 17.9 mmol, 1.2 equiv.) was added to a solution of 6-7 (7.5 g, 14.9 mmol) in pyridine (75 mL) under Ar. The resulting solution was stirred at room temperature for 12 hours. The reaction mixture was quenched with MeOH (10 mL), diluted with dichloromethane (10 mL), washed with saturated aqueous sodium bicarbonate solution 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: 1-40: 1) to give 8 g (66% yield) of 6-8 as a light yellow oil. MS: m / z 804.45 [MH] - . 1 H NMR(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).

[0462] Preparation of 6

[0463] To a solution of 6-8 (8 g, 9.9 mmol) in dichloromethane (80 mL) was added 3-((bis(diisopropylamino)phosphinoyl)oxy)propionitrile (4.2 g, 13.9 mmol, 1.4 equiv) and 4,5-dicyanoimidazole (1.4 g, 11.9 mmol, 1.2 equiv) under Ar. The reaction mixture was stirred at room temperature for 1 hour, then diluted with dichloromethane (80 mL), washed with saturated aqueous sodium bicarbonate solution 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 and THF containing 0.05% NH4HCO3; 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.

[0464] Example 7 .

[0465]

[0466] Preparation of 7-1

[0467] At -20 ° C, under N2, MeOH (4g, 125.3mmol, 10 equivalents), NIS (3.1g, 13.8mmol, 1.1 equivalents) and trimethylsilyl trifluoromethanesulfonate (557mg, 2.5mmol, 0.2 equivalents) were added to a solution of 6-1 (6g, 12.5mmol) in dichloromethane (180mL). The resulting solution was stirred at -20 ° C for 1 hour and then quenched with aqueous sodium thiosulfate solution (300mL). The mixture was extracted with dichloromethane (3×600mL), and the combined organic extracts were dried (anhydrous Na2SO4), filtered and concentrated. The residue was purified on a silica gel column with ethyl acetate / petroleum ether (1:50 to 1:1) to obtain 4g (72% yield) of 7-1 as a yellow oil.

[0468] Preparation of 7-2

[0469] At 0 ° C, NaOMe (1 g, 18.6 mmol, 1.2 equiv) was added to a solution of 7-1 (6 g, 15.5 mmol) in MeOH (100 mL) under Ar and stirred at 0 ° C for 0.5 hours. 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: 10-5: 1) to give 4.5 g (84%) of 7-2 as a yellow oil. MS: m / z 386.17 [M+H] +.

[0470] Preparation of 7-3

[0471] At 0 ° C, sodium hydride (60% in mineral oil, 2.6 g) was added to a solution of 7-2 (4.5 g, 13.1 mmol) in DMF (90 mL), and the resulting mixture was stirred for 15 minutes. 1-Iodohexadecane (9.21 g, 26.1 mmol, 2.0 equiv) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with water (200 mL) and extracted with dichloromethane (3×500 mL). The combined organic fractions were dried (anhydrous Na2SO4), filtered and concentrated. The residue was purified on a silica gel column with ethyl acetate / petroleum ether (1:50-1:1) to give 5.5 g (74% yield) of 7-3 as a yellow oil. MS: m / z 568.41 [M+Na] + . 1H 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).

[0472] Preparation of 7-4

[0473] 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 H2 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. It was used in the next step without further purification. MS: m / z 388.32 [M+H] + .

[0474] Preparation of 7-5

[0475] To a solution of 7-4 (3.2 g, 8.2 mmol) in pyridine (64 mL) was added 4,4'-(chloro(phenyl)methylene)bis(methoxybenzene) (2.93 g, 8.6 mmol, 1.1 equiv) at 0 °C under N2. The resulting solution was stirred at room temperature for 2 hours, then diluted with dichloromethane (200 mL) and washed with water. The organic layer was dried (anhydrous Na2SO4), filtered and concentrated. The residue was purified on a silica gel column with 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).

[0476] Preparation of 7

[0477] 3-((bis(diisopropylamino)phosphinoyl)oxy)propionitrile (1.7 g, 5.6 mmol, 1.3 equiv) and 4,5-dicyanoimidazole (564 mg, 4.8 mmol, 1.1 equiv) were added to a solution of 7-5 (3 g, 4.3 mmol) in dichloromethane (30 mL), and the resulting solution was stirred at room temperature for 1 hour. 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 with 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.

[0478] Example 8 .

[0479]

[0480] Preparation of 8-1

[0481] At -20 ° C, under an inert atmosphere of argon, 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) was added N-iodosuccinimide (37 g, 151.1 mmol, 1.1 equiv) and silver trifluoromethanesulfonate (4 g, 15.06 mmol, 0.1 equiv). The resulting solution was stirred at -20 ° C for 30 minutes. 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: 1-20: 1) to give 70 g (79%) of 8-1 as a light 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).

[0482] Preparation of 8-2

[0483] At 0 ° C, under an inert atmosphere of argon, 35 mL of 5M methanol solution of sodium methoxide was added to a solution of 8-1 (70 g, 117.4 mmol, 1.0 equivalent) and a mixture dissolved in 700 mL of methanol. The resulting solution was stirred at 0 ° C for 30 minutes. The resulting mixture was quenched with 40 mL of 3M hydrochloric acid and concentrated under reduced pressure. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (80: 1-50: 1) to give 51 g (78%) of 8-2 as a light yellow oil. MS: m / z 577.45 [M+Na] + .

[0484] Preparation of 8-3

[0485] At 0 ° C, sodium hydride (11 g, 460 mmol, 5.0 equivalents) was added to a solution of 8-2 (51 g, 92.1 mmol) in 510 mL of DMF. The resulting solution was stirred at 0 ° C for 30 minutes. Then 1-iodohexadecane (47 g, 138 mmol, 1.5 equivalents) was added at 0 ° C and stirred at room temperature for 16 hours. The reaction mixture was quenched with 200 mL of saturated ammonium chloride solution, diluted with 3000 mL of ethyl acetate, washed with 2 × 1800 mL of water, 1 × 1800 mL of saturated sodium thiosulfate aqueous solution, saturated sodium bicarbonate aqueous solution and saturated sodium chloride aqueous solution. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (100: 1-80: 1) to give 49 g (76%) of 8-3 as a yellow solid. MS: m / z 796.85 [M + NH4] + .

[0486] Preparation of 8-4

[0487] At room temperature, 10% Pd / C (49 g) and acetic acid (50 mL) were added 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 and stirred for 12 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure. 32 g (86% yield) of 8-4 was obtained as a white solid. It was used in the next step without further purification. MS: m / z 630.65 [M+NH4] + .

[0488] Preparation of 8-5

[0489] Under an inert atmosphere of argon, 4,4'-dimethoxytrityl chloride (20g, 58.8mmol, 1.2 equivalents) was added to a solution of 8-4 (30g, 48.9mmol) in 300mL of pyridine and stirred at room temperature for 3 hours. The reaction mixture was quenched with 150mL of methanol, diluted with 2500mL of dichloromethane, washed with 2×2000mL of saturated aqueous sodium bicarbonate solution and 2×2000mL of saturated aqueous sodium chloride solution. 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 containing 0.05% TEA (60: 1-30: 1). 8-5 was obtained as a light yellow oil of 39.1g (88% yield). MS: m / z 899.60 [MH] - . 1H 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).

[0490] 8. Preparation

[0491] At room temperature, 3-(bis(diisopropylamino)phosphinoyl)propionitrile (14 g, 46.7 mmol, 1.2 equiv.), 4,5-dicyanoimidazole (5 g, 42.8 mmol, 1.1 equiv.) were added to a solution of 8-5 (35 g, 38.9 mmol) in 350 mL of dichloromethane under an inert atmosphere of argon. The resulting solution was stirred for 1 hour, then diluted with 500 mL of dichloromethane, washed with 3×2000 mL of saturated aqueous sodium bicarbonate solution and 3×2000 mL of saturated aqueous sodium chloride solution. 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 containing 0.05% TEA (80:1-30:1) to give 21 g (51% yield) of compound 8 as a colorless oil. MS: m / z 1101.80 [M+H] + . 1H 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.

[0492] Example 9 .

[0493]

[0494] Preparation of 9-1

[0495] At 0 ° C, iodine was added to a solution of (3R, 4R, 5R) -5- (acetoxymethyl) tetrahydrofuran -2,3,4- triyl triacetate (100g, 314.2mmol) in 1000mL acetone under an inert atmosphere of argon. The resulting solution was stirred at room temperature for 3.5 hours. The reaction mixture was diluted with 2000mL ethyl acetate and washed with 2×500mL saturated sodium thiosulfate aqueous solution and 2×500mL saturated sodium bicarbonate aqueous solution, respectively. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in 700mL methanol and cooled to 0 ° C. Then 54mL of 5M sodium methoxide methanol solution was added and stirred at 0 ° C for 30 minutes under an inert atmosphere of argon. The reaction mixture was quenched with 90mL 3N hydrochloric acid and concentrated under reduced pressure. The crude product was applied to a silica gel column and eluted with dichloromethane / methanol (120:1-40:1) to give 37 g (62%) of 9-1 as a white solid. MS: m / z 189.05 [MH] - .

[0496] Preparation of 9-2

[0497] At 0 ° C, sodium hydride (8.34 g, 347.4 mmol, 3.0 equivalents) was slowly added to a solution of 9-1 (22 g, 115.8 mmol) in anhydrous DMF (220 mL) under an inert atmosphere of argon, followed by dropwise addition of benzyl bromide (49.5 g, 289.5 mmoL, 2.5 equivalents) at 0 ° C. The resulting solution was stirred at room temperature for 3 hours. 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 sodium chloride aqueous solution. The crude product was loaded onto a silica gel column and eluted with petroleum ether / ethyl acetate (80: 1-20: 1) to give 35.2 g (82% yield) of 9-2 as a yellow oil. MS: m / z 393.25 [M+Na] + . 1 H 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).

[0498] Preparation of 9-3

[0499] A solution of 9-2 (30g, 81.1mmol) in 150mL trifluoroacetic acid and 150mL water was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in 300mL pyridine and cooled to 0°C. Then 60mL acetic anhydride was added and stirred at room temperature for 3 hours. The reaction mixture was quenched with 60mL water and concentrated under reduced pressure. The residue was diluted with 1000mL ethyl acetate and washed with 2×500mL water, 1×200mL 2N hydrochloric acid, 1×400mL saturated sodium bicarbonate aqueous solution, 2×500mL water and 2×400mL saturated sodium chloride aqueous solution. 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 (20:1-8:1) to give 21.5g (64%) of 9-3 as a light yellow oil. MS: m / z 437.20 [M+Na] + .

[0500] Preparation of 6-1

[0501] At 0 ° C, 10 mL of 1M tin chloride pentahydrate in dichloromethane was added to a solution of 9-3 (21.5 g, 51.9 mmol) and (7.7 g, 62.3 mmoL, 1.2 equivalents) in dichloromethane (215 mL) under an inert atmosphere of argon, and stirred at room temperature for 1 hour. 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: 1-20: 1) to give 16 g (64%) of 6-1 as a light yellow oil. MS: m / z 496.20 [M+Na] + .

[0502] Preparation of 9-5

[0503] At -20 ° C, under an inert atmosphere of argon, to a solution of 6-1 (5 g, 10.5 mmol) and 1-heptadecanol (3.2 g, 12.6 mmol, 1.2 equiv) in dichloromethane (50 mL) was added N-iodosuccinimide (2.6 g, 11.5 mmol, 1.1 equiv) and silver trifluoromethanesulfonate (270 mg, 1.1 mmol, 0.1 equiv). The resulting solution was stirred at -20 ° C for 30 minutes. The reaction mixture was quenched with 1 mL of trimethylamine and concentrated under reduced pressure. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (50: 1-20: 1) to give 4.3 g (67% yield) of 9-5 as a light yellow oil. MS: m / z 633.45 [M+Na] + .

[0504] Preparation of 9-6

[0505] At 0 ° C, 1.4 mL of 5M sodium methoxide in methanol was added to a solution of 9-5 (4.30 g, 7.1 mmol) in methanol (43 mL) under Ar. The resulting solution was stirred at 0 ° C for 30 minutes, then the reactant was quenched with hydrochloric acid (3 M, 2 mL) and concentrated. The crude product was applied to 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 light yellow oil. MS: m / z 591.45 [M+Na] + . 1H 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).

[0506] Preparation of 9-7

[0507] At 0 ° C, sodium hydride (850 mg, 35.2 mmol, 5.0 equivalents) was added to a solution of 9-6 (4 g, 7.04 mmol) in 40 mL of DMF. The resulting solution was stirred at 0 ° C for 30 minutes. Then 1-iodohexadecane (3.7 g, 10.56 mmol, 1.5 equivalents) was added at 0 ° C and stirred at room temperature for 16 hours. The reaction mixture was quenched with 10 mL of saturated ammonium chloride solution, diluted with 300 mL of ethyl acetate, washed with 2 × 150 mL of water, 1 × 150 mL of saturated sodium thiosulfate aqueous solution and saturated sodium bicarbonate aqueous solution and 1 × 150 mL of saturated sodium chloride aqueous solution. The crude product was applied to a silica gel column and eluted with petroleum ether / ethyl acetate (100: 1-80: 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).

[0508] Preparation of 9-8

[0509] At room temperature, 10% Pd / C (410 mg) and acetic acid (4 mL) were added to a solution of 9-7 (4.1 g, 5.2 mmol) in 32 mL of tetrahydrofuran and 8 mL of methanol under a hydrogen atmosphere. The reaction mixture was stirred for 12 hours, then filtered, and the filtrate was concentrated under reduced pressure to give 2.3 g (73%) of 9-8 as a white solid. It was used in the next step without further purification. MS: m / z 630.65 [M+NH4] + .

[0510] Preparation of 9-9

[0511] At room temperature, 4,4'-dimethoxytrityl chloride (1.58 g, 4.1 mmol, 1.2 equiv.) was added to a solution of 9-8 (2.1 g, 3.4 mmola) in 21 mL of pyridine under an inert atmosphere of argon. The resulting solution was stirred at 25 °C for 3 hours. The reaction mixture was quenched with 10 mL of methanol, diluted with 200 mL of dichloromethane, washed with 2×150 mL of saturated aqueous sodium bicarbonate solution and 2×150 mL of saturated aqueous sodium chloride solution. 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 containing 0.05% TEA (60:1-30:1) to give 2.3 g (73% yield) of 9-9 as a light yellow oil. MS: m / z 913.60 [MH] - . 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).

[0512] Preparation of 9

[0513] At room temperature, 3-(bis(diisopropylamino)phosphinoyl)propionitrile (910 mg, 3.0 mmol, 1.2 equiv) and 4,5-dicyanoimidazole (330 mg, 2.8 mmol, 1.1 equiv) were added to a solution of 9-9 (2.3 g, 2.5 mmol) in dichloromethane (23 mL) under argon and stirred at room temperature for 1 hour. The resulting solution was diluted with 500 mL of dichloromethane, washed with 3×300 mL of saturated aqueous sodium bicarbonate solution and 3×300 mL of saturated aqueous sodium chloride solution. 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 with hexane / ethyl acetate containing 0.05% TEA (80:1-30:1) to give 2.04 g (73% yield) of compound 9 as a light 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.

[0514] Example 10 .

[0515]

[0516] Preparation of 10-1

[0517] At 0°C, sodium hydride (60% dispersion in oil, 1.7 g, 43.9 mmoL, 5.0 equiv) was added to a solution of 9-6 (5 g, 8.8 mmol) in 50 mL of N,N-dimethylformamide and stirred at 0°C for 30 minutes. 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 hours. 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 saturated brine and dried over anhydrous sodium sulfate. The evaporated crude residue was purified on a silica gel column with dichloromethane / methanol (100 / 1 to 80 / 1) to give 5 g (90% yield) of 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).

[0518] Preparation of 10-2

[0519] Under hydrogen atmosphere, 10% Pd / C (500 mg) and acetic acid (5 mL, 87.4 mmol) were added to a solution of 10-1 (5 g, 7.9 mmol) in methanol (50 mL) and tetrahydrofuran (50 mL). The resulting mixture was stirred at room temperature for 8 hours, then filtered and the filtrate was concentrated. The crude product was purified on a silica gel column with 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).

[0520] Preparation of 10-3

[0521] At 0 ° C, 4,4'-dimethoxytrityl chloride (2.4 g, 7.2 mmoL, 1.2 equivalents) was added to a solution of 10-2 (2.7 g, 6.0 mmol) in pyridine (27 mL) under an inert atmosphere of argon. The resulting solution was stirred at 25 ° C for 6 hours. The reaction mixture was quenched with 10 mL of methanol, diluted with 100 mL of dichloromethane, washed with 2 × 100 mL of saturated sodium bicarbonate solution and 2 × 100 mL of saturated brine. The organic layer was dried (Na2SO4), filtered and concentrated. The crude product was purified on a silica gel column with 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 / z747.45 [MH] - . 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).

[0522] 10. Preparation

[0523] To a solution of 10-3 (2.5 g, 3.3 mmol) in 24 mL of dichloromethane was added 3-((bis(diisopropylamino)phosphinoyl)oxy)propionitrile (1.13 g, 3.96 mmoL, 1.2 equiv.), 4,5-dicyanoimidazole (427 mg, 3.63 mmoL, 1.1 equiv.) at room temperature. The solution was stirred at 25 °C for 45 minutes. The resulting mixture was diluted with 100 mL of dichloromethane, washed with 2×100 mL of saturated sodium bicarbonate solution and 2×100 mL of saturated brine. The organic layer was dried (Na2SO4), filtered and concentrated. The crude product was purified on a silica gel column with hexane / ethyl acetate (20 / 1 to 6 / 1). The fractions containing the product were concentrated to give 2.0 g (63% yield) of compound 10 as a light 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.

[0524] Embodiment 11 .

[0525]

[0526] Preparation of 11-1

[0527] At 0 ° C, tetrabutylammonium hydrogen sulfate (11.5 g, 34.0 mmol, 0.2 equiv.), 1M NaOH (850 mL) and bromoacetonitrile (61.3 g, 510.7 mmol, 3.0 equiv.) were added to a solution of (4aR, 7R, 8R, 8aS)-6-methoxy-2-phenylhexahydropyrano[3,2-d][1,3]dioxin-7,8-diol (48 g, 170 mmol) in dichloromethane (480 mL). The resulting mixture was stirred at room temperature for 2 hours, then diluted with dichloromethane (400 mL), 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 with 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, 4.1 Hz, 1H), 4.04 (t, J = 9.1 Hz, 1H), 3.92-3.73 (m, 2H), 3.72-3.61 (m, 2H), 3.49 (s, 3H).

[0528] Preparation of 11-2

[0529] At 0°C, LiAlH4 (4.2 g, 111.1 mmol, 2.0 equiv.) was added to a solution of 11-1 (20 g, 55.6 mmol) in THF (200 mL) under Ar. The resulting mixture was stirred at 0°C for 1 hour and then quenched with EtOAc. The solid was filtered out. The filtrate was diluted with dichloromethane, 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 with 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] + .

[0530] Preparation of 11-3

[0531] To a solution of 11-2 (10 g, 27.1 mmol) in dichloromethane (200 mL) at 0°C under Ar was added 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). The mixture was stirred at room temperature for 16 h, then diluted with dichloromethane, 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 with 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] + .

[0532] Preparation of 11-4

[0533] 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 hours, 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] + .

[0534] Preparation of 11-5

[0535] To a solution of 11-4 (4.5 g, 5.94 mmol) in CH2Cl2 (180 mL) was added 4,4'-dimethoxytrityl chloride (2.2 g, 6.53 mmol, 1.10 equiv), Et3N (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 hours and quenched with methanol. The mixture was then diluted with dichloromethane, washed with saturated aqueous NaHCO3 solution and saturated 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 (60:1 to 30:1) to give 3.2 g (50%) of 11-5 as a light yellow oil. MS: m / z 1081.85 [M+Na] + . 1H 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).

[0536] Preparation of 11

[0537] At room temperature, 2-cyanoethyl-3-((bis(diisopropylamino)phosphinoyl)oxy)propionitrile (1.2 g, 3.96 mmol, 1.4 equiv) and 4,5-dicyanoimidazole (401 mg, 3.39 mmol, 1.2 equiv) were added to a solution of 11-5 (3 g, 2.83 mmol) in dichloromethane (30 mL) under Ar. The reaction mixture was stirred for 1 hour, then diluted with dichloromethane, washed with saturated NaHCO3 aqueous solution and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on flash preparative HPLC using the following conditions (column, C18; mobile phase, water (containing 0.05% NH4HCO3) and tetrahydrofuran (20% tetrahydrofuran to 100% in 10 minutes, and 100% tetrahydrofuran maintained for 5 minutes). The aqueous layer was separated and extracted with dichloromethane. The combined organic layers were combined and dried over anhydrous Na2SO4, 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.

[0538] Example 12.

[0539]

[0540] Preparation of 12-1

[0541] At 0 ° C, sodium hydride (60% dispersion in mineral oil) (18.4 g, 461.53 mmol, 1.5 equivalents) was added to a solution of diacetone-D-glucose (80 g, 307.35 mmol) in tetrahydrofuran (800 mL) under N2. The resulting solution was stirred for 20 minutes, iodomethane (65.4 g, 461.53 mmol, 1.5 equivalents) was added at 0 ° C, and stirred at room temperature overnight. The reactants were quenched with saturated NH4Cl solution at 0 ° C 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 with 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] + .

[0542] Preparation of 12-2

[0543] A solution of 12-1 (70 g, 255.474 mmol) in 4M HCl (700 mL) was stirred at room temperature for 1 hour and then concentrated. The residue was dissolved in pyridine (550 mL) at 0 ° C under N2, 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 with petroleum ether / ethyl acetate (30 / 1 to 5 / 1) to obtain 69 g (75% yield) of 12-2 as a yellow solid. MS: m / z 363.15 [M+H] + .

[0544] Preparation of 12-3

[0545] At 0 ° C, 1-hexadecanol (50.1 g, 207.18 mmol, 1.5 equivalents) was added to a solution of 12-2 (50 g, 138.12 mmol) in dichloromethane (750 mL) under N2. The resulting solution was stirred for 20 minutes, and boron trifluoride etherate (58.8 g, 414.36 mmol, 3 equivalents) was added dropwise and stirred at room temperature for 3 hours. The reactant was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layer was washed with saturated brine, dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (30 / 1 to 5 / 1) to obtain 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).

[0546] Preparation of 12-4

[0547] At 0 ° C, sodium methoxide (12.8 g, 321.69 mmol, 5 equivalents) was added to a solution of 12-3 (35 g, 64.33 mmol) in methanol (350 mL) under N2 and stirred at room temperature for 2 hours. 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 with 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.

[0548] Preparation of 12-5

[0549] At 0 ° C, to a solution of 12-4 (18 g, 43.06 mmol, 1.0 equiv) in chloroform (180 mL) under N2, (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 hours. The reaction mixture was concentrated and purified on a silica gel column with 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] + .

[0550] Preparation of 12-6

[0551] At 0 ° C, sodium hydride (60% dispersion in mineral oil) (1.5 g, 38.53 mmol, 1.5 equiv.) was added to a solution of 12-5 (13 g, 25.69 mmol) in tetrahydrofuran (200 mL) under N2. The resulting solution was stirred at 0 ° C for 30 minutes, 1-iodohexadecane (18 g, 51.38 mmol, 2.0 equiv.) was added and the reactants were stirred at room temperature for 2 hours. The reactants were 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 with 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] + .

[0552] Preparation of 12-7

[0553] At room temperature, 10% palladium / activated carbon (11 g) was added to a solution of 12-6 (11 g, 15.06 mmol, 1.0 equivalent) in tetrahydrofuran (100 mL) and dichloromethane (100 mL). After five cycles of flushing the reaction mixture with H2, the resulting solution was stirred at room temperature for 4 hours, then filtered and concentrated. The residue was purified on a silica gel column with 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] + .

[0554] Preparation of 12-8

[0555] To a solution of 12-7 (7.7 g, 11.99 mmol, 1 eq.) in dichloromethane (100 mL) at 0 °C under N2 was added triethylamine (1.8 g, 17.99 mmol, 1.5 eq.), 4-dimethylaminopyridine (146.3 mg, 1.19 mmol, 0.1 eq.) and 1-[chloro(4-methoxyphenyl)benzyl]-4-methoxybenzene (4.8 g, 14.39 mmol, 1.2 eq.). The resulting solution 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 with petroleum ether / ethyl acetate (30 / 1 to 5 / 1, containing 0.5% triethylamine) to give 12-8 with a purity of 94%. To further improve the purity, it was recrystallized by dissolving it in dichloromethane (1 mL / g), and added dropwise to acetonitrile (20 mL / g), followed by filtration to obtain 6 g (58% yield) of 12-8 with a purity of 97% as a white solid. MS: m / z 943.55 [MH] - . 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).

[0556] Preparation of 12

[0557] At room temperature, bis(diisopropylamino)(2-cyanoethoxy)phosphine (2.2g, 7.38mmol, 1.2 equivalents) and 4,5-dicyanoimidazole (800mg, 6.77mmol, 1.1 equivalents) were added to a solution of 12-8 (6g, 6.15mmol) in dichloromethane (60mL) under N2 and stirred for 1 hour. The reaction mixture was diluted with dichloromethane and washed with saturated NaHCO3 aqueous solution and saturated brine. The organic layer was dried over anhydrous Na2OS4, filtered and concentrated. The residue was purified on a fast 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 with 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.

[0558] Embodiment 13 .

[0559]

[0560] Preparation of 13-1

[0561] At 0 ° C, sodium hydride (60%, 24.6 g, 615.4 mmol, 2.0 equiv) was added to a solution of (3aR, 5S, 6S, 6aR) -5- ((R) -2,2- dimethyl -1,3- dioxol -4- yl) -2,2- dimethyltetrahydrofuran [2,3-d] [1,3] dioxol -6- alcohol (80 g, 307.6 mmol) in DMF (800 mL) under N2 and stirred for 15 minutes. 1-Iodohexadecane (162 g, 461.4 mmol, 1.5 equiv) was added to the reaction mixture and stirred for 2 hours at room temperature. The reaction mixture was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layers were washed with water and saturated brine, and then the organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on a silica gel column with 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] + .

[0562] Preparation of 13-2

[0563] To a solution of 13-1 (110 g, 227.2 mmol) in tetrahydrofuran (1100 mL) at 0 ° C., 6 M HCl (378 mL, 2.27 mol, 10 equiv.) was added. The resulting solution was stirred at 40 ° C. for 12 hours and then quenched with pyridine (201 mL, 2.5 mol, 11 equiv.). The resulting solution was concentrated and co-evaporated with dry pyridine to give 110 g (crude product) of 13-2 as a yellow oil. It was used in the next step without further purification. MS: m / z 422.35 [M+NH4] + .

[0564] Preparation of 13-3

[0565] At 0 ° C, acetic anhydride (550 mL) was added to a solution of 13-2 (110 g, crude product) in pyridine (1100 mL) under N2 and stirred for 16 hours. 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 with petroleum ether / ethyl acetate (20 / 1 to 8 / 1) to obtain 100 g (70% yield, over two steps) of 13-3 as a yellow oil. MS: m / z 590.45 [M+NH4] + .

[0566] Preparation of 13-4

[0567] At 0 ° C, 13-3 (100 g, 104.8 mmol) and 1-hexadecanol (55.3 g, 157.2 mmol, 1.5 equivalents) in dichloromethane (215 mL) solution was added under Ar 1M tin (IV) chloride in dichloromethane (314 mL) solution and stirred at room temperature for 1 hour. The reaction mixture was quenched with trimethylamine and concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (60: 1 to 20: 1) to give 51 g (64% yield) of 13-4 as a light 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).

[0568] Preparation of 13-5

[0569] At 0 ° C, at 0 ° C, under Ar, to a solution of 13-4 (50 g, 66.26 mmol) in methanol (500 mL) was added 5M sodium methoxide in methanol (20 mL) and stirred at room temperature for 2 hours. The resulting mixture was quenched with 3M hydrochloric acid (20 mL) and concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (50: 1 to 1: 1) to give 25 g (80% yield) of 13-5 as a light yellow oil. MS: m / z 646.60 [M+NH4] + .

[0570] Preparation of 13-6

[0571] At 0 ° C, benzaldehyde dimethyl acetal (9 g, 59.71 mmol, 1.5 equivalents) and copper (II) trifluoromethanesulfonate (2.8 g, 7.96 mmol, 0.2 equivalents) were added to a solution of 13-5 (25 g, 39.80 mmol) in chloroform (250 mL) under N2 and stirred at room temperature for 16 hours. The reaction mixture was quenched with trimethylamine and concentrated. The residue was purified on a silica gel column with 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).

[0572] Preparation of 13-7

[0573] At 0 ° C, sodium hydride (60%, 1.9 g, 47.48 mmol, 2.0 equivalents) was added to a solution of 13-6 (17 g, 23.74 mmol) in DMF (170 mL) under N2 and stirred for 15 minutes. Iodomethane (5 g, 35.61 mmol, 1.5 equivalents) was added to the reaction mixture and stirred at room temperature for 2 hours. 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 with petroleum ether / ethyl acetate (20 / 1 to 8 / 1) to obtain 12 g (77% yield) of 13-7 as a yellow oil. MS: m / z 731.55 [M+H] + .

[0574] Preparation of 13-8

[0575] 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 H2 for five cycles. The resulting solution was stirred at room temperature for 16 hours, then filtered and concentrated to give 9 g (80% yield) of 13-8 as a white solid. MS: m / z 660.65 [M+NH4] + .

[0576] Preparation of 13-9

[0577] At 0 ° C, 4,4'-dimethoxytrityl chloride (5.6 g, 16.8 mmoL, 1.2 equiv.), Et3N (3.9 mL, 28 mmol) and DMAP (340 mg, 2.8 mmol) were added to a solution of 13-8 (9 g, 13.99 mmol) in CH2Cl2 (500 mL) under Ar and stirred at 50 ° C and room temperature for 4 hours. The reaction mixture was quenched with methanol, diluted with dichloromethane, washed with saturated NaHCO3 and saturated 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 (40 / 1 to 2 / 1) to give 6 g (75% yield) of 13-9 as a colorless oil. MS: m / z 943.65 [MH] - . 1 H 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),3.92-3.6 2(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).

[0578] Preparation of 13

[0579] At room temperature, 3-(bis(diisopropylamino)phosphinoyl)propionitrile (2.3 g, 7.62 mmol, 1.2 equiv) and 4,5-dicyanoimidazole (824 mg, 6.98 mmol, 1.1 equiv) were added to a solution of 13-9 (6 g, 6.35 mmol) in dichloromethane (60 mL) under Ar and stirred for 1 hour. The resulting solution was diluted with dichloromethane, washed with saturated NaHCO3 aqueous solution and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on Prep-Flash using the following conditions (C-18 column, mobile phase, water and tetrahydrofuran, gradient 30% to 100% THF). The fraction containing the product was diluted with an equal volume of dichloromethane, and the aqueous layer was separated and extracted with dichloromethane. The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on a silica gel column with 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] + . 1 H 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.

[0580] Embodiment 14 .

[0581]

[0582] Preparation of 14-1

[0583] At 0 ° C, sodium hydride (60%, 3g, 70.92mmol, 4.0 equivalents) was added to a solution of (4aR, 7R, 8R, 8aS)-6-methoxy-2-phenylhexahydropyran and [3,2-d] [1,3] dioxin-7,8-diol (5g, 17.73mmol) in DMF (50mL) under N2 and stirred for 15 minutes. 1-Iodohexadecane (12g, 35.46mmol, 2.0 equivalents) was added to the reaction mixture and stirred at room temperature for 2 hours. 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 residue was purified on a silica gel column with ethyl acetate / petroleum ether (1 / 50 to 1 / 1) to obtain 5g (50% yield) of 14-1 as a white solid. MS: m / z 731.60[M+H] + .

[0584] Preparation of 14-2

[0585] To a solution of 14-1 (5 g, 6.84 mmol, 1.0 eq.) in tetrahydrofuran (50 mL) was added 10% palladium / activated carbon (wt 200 mL) at room temperature. t / w t =100%, 5 g) and flushed with H2 for five cycles. The resulting solution was stirred at room temperature for 16 hours, 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] + .

[0586] Preparation of 14-3

[0587] At 0 ° C, 4,4'-(chloro(phenyl)methylene)bis(methoxybenzene) (2.3 g, 7.08 mmol, 1.3 equiv) was added to a solution of 14-2 (3.5 g, 5.45 mmol, 1 equiv) in pyridine (40 mL) under N2 and stirred at room temperature for 2 hours. The reaction mixture was diluted with dichloromethane and washed with saturated NaHCO3 aqueous solution and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on a silica gel column with 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 [MH] - . 1H 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).

[0588] Preparation of 14

[0589] To a solution of 14-3 (4 g, 4.23 mmol, 1.0 equiv) in dichloromethane (40 mL) was added 3-(bis(diisopropylamino)phosphinoyl)propionitrile (1.5 g, 5.07 mmol, 1.2 equiv) and 4,5-dicyanoimidazole (549 mg, 4.65 mmol, 1.1 equiv) at 0°C under Ar and stirred at room temperature for 1 hour. The reaction mixture was diluted with dichloromethane, washed with saturated aqueous NaHCO3 and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on Prep-Flash using the following conditions: column, C18 silica gel; mobile phase, water and tetrahydrofuran (30% tetrahydrofuran to 100% in 15 minutes and maintained at 100% for 5 minutes); detector, UV 254 nm. The fractions were 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 with 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] + . 1H 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.

[0590] Example 15.

[0591]

[0592] Preparation of 15-1

[0593] At 0 ° C, 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) were added 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). The solution was stirred at room temperature for 12 hours and then diluted with dichloromethane. The organic layer was washed with water and saturated brine, then dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on a silica gel column with 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+H2O] + .

[0594] Preparation of 15-2

[0595] Triethylamine (4.6 g, 45.1 mmoL, 20 equiv) and 15-1 (2.6 g, 2.3 mmoL, 1 equiv) were added to a solution of triethylamine trihydrofluoride (3.6 g, 22.6 mmoL, 10 equiv) in tetrahydrofuran (52 mL) at room temperature. The solution was stirred at room temperature for 12 hours, 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] + .

[0596] Preparation of 15-3

[0597] At 0 ° C, 4,4'-(chloro(phenyl)methylene)bis(methoxybenzene) (447 mg, 1.3 mmoL, 1.2 equiv.), triethylamine (222 mg, 2.2 mmoL, 2 equiv.) and 4-dimethylaminopyridine (13 mg, 0.1 mmoL, 0.1 equiv.) were added to a solution of 15-2 (1 g, 1.1 mmol) in dichloromethane (100 mL) under N2 and stirred at 50 ° C for 1 hour. The reaction mixture was precipitated by acetonitrile and filtered. The residue was purified on a silica gel column with 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 [MH] - .

[0598] Preparation of 15

[0599] 3-((bis(diisopropylamino)phosphinoyl)oxy)propionitrile (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 to a solution of 15-3 (2.8 g, 2.3 mmol) in dichloromethane (56 mL), and stirred at room temperature for 45 minutes. The reaction mixture was diluted with dichloromethane and washed with saturated aqueous NaHCO3 solution and saturated brine. The organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by C18 flash chromatography (mobile phase, THF aqueous solution (0.05% ammonium bicarbonate), 20% to 100% gradient in 25 minutes). The fractions were diluted with an equal volume of dichloromethane. The organic phase 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 obtain 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.

[0600] Example 16 .

[0601]

[0602] Preparation of 16-1

[0603] At 0 ° C, sodium hydride (5g, 127.5mmol, 1.1 equivalents) was added to a solution of (tert-butoxycarbonyl) glycine methyl ester (22g, 116.40mmol) in 220mL N,N-dimethylformamide. The resulting solution was stirred at 0 ° C for 30 minutes. Then 1-iodohexadecane (49g, 139.58mmol, 1.2 equivalents) was added at 0 ° C and stirred at room temperature for 16 hours. The reaction mixture was quenched with 50mL saturated ammonium chloride solution, diluted with 1500mL ethyl acetate, washed with 2×300mL water, 1×300mL saturated sodium thiosulfate aqueous solution and saturated sodium bicarbonate aqueous solution and 1×300mL saturated brine. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (70 / 1 to 20 / 1) to give 22g (52% yield) of 16-1 as a yellow solid. MS: m / z 436.35 [M+Na] + .

[0604] Preparation of 16-2

[0605] 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 hour. The solvent was removed under reduced pressure, and the crude product was purified on a silica gel column with 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] + .

[0606] Preparation of 16-3

[0607] Palmitic acid (13 g, 48.86 mmol, 1.2 equiv) was added 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 at 0°C under an inert atmosphere of argon. The mixture was stirred for 10 minutes, 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 hours, then diluted with 1000 mL of dichloromethane and washed with 2×300 mL of water and 2×300 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified on a silica gel column with hexane / ethyl acetate (70 / 1 to 2 / 1) to give 13 g (58% yield) of 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).

[0608] Preparation of 16-4

[0609] At 0 ° C, 5M lithium hydroxide (24 mL, 119.55 mmol, 5.0 equivalents) was added to a solution of 16-3 (13 g, 23.91 mmol) in 130 mL of tetrahydrofuran under an inert atmosphere of argon. The resulting solution was stirred at room temperature for 3 hours. The mixture was diluted with 1000 mL of dichloromethane, and the pH was adjusted to 6 with 2M hydrochloric acid, and then washed with 2×300 mL of water and 2×300 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was applied to a silica gel column and purified with 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] + .

[0610] Preparation of 16-5

[0611] At 0 ° C, methylglycine tert-butyl ester (4 g, 23.6 mmol, 1.2 equiv) was added 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 under an inert atmosphere of argon. The mixture was stirred for 10 minutes, 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 hours, then diluted with 1000 mL of dichloromethane and washed with 2×300 mL of water and 2×300 mL of saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was applied to 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] + .

[0612] Preparation of 16-6

[0613] 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 hours. The solvent was removed under reduced pressure, and the crude product was purified on a silica gel column with 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] + .

[0614] Preparation of 16

[0615] At 0 ° C, 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 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. 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).

[0616] Embodiment 17 .

[0617]

[0618] Preparation of 17-1

[0619] At room temperature, hydrogen bromide (20.9 g, 257.827 mmol, 1.1 equivalents) was added to a solution of tetradecane-1,14-diol (54 g, 234.388 mmol) in 540 mL of cyclohexane, and the solution was stirred at 80 ° C for 16 hours. The reactant was quenched by adding a saturated sodium bicarbonate solution (500 mL), and the resulting solution was extracted with 3 × 800 mL of dichloromethane. The combined organic layer was washed with 2 × 500 mL of water and 500 mL of brine. 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 (100 / 1 to 4 / 1) to give 42 g (61% yield) of 17-1 as a white solid.

[0620] Preparation of 17-2

[0621] At 0 ° C, sodium hydride (3.6 g, 150.02 mmol, 2.0 equivalents) was added to a solution of 17-1 (22 g, 75.0 mmol) in 440 mL of dimethylformamide, and the solution was stirred at 0 ° C for 30 minutes, and then (bromomethyl) benzene (19.2 g, 112.5 mmol, 1.5 equivalents) was added at 0 ° C. The solution was then stirred at room temperature for 18 hours. The reactant was quenched by adding a saturated ammonium chloride solution (500 mL), and the resulting solution was extracted with 3 × 500 mL of dichloromethane. The combined organic layer was washed with 2 × 300 mL of water and 300 mL of saturated brine. 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 with petroleum ether / ethyl acetate (100 / 1 to 4 / 1) to give 21.0 g (73% yield) of 17-2 as a yellow oil.

[0622] Preparation of 17-3

[0623] Triphenylphosphine (68.4 g, 260.8 mmol, 2.0 equiv.) was added to a solution of 17-2 (50 g, 130.4 mmol) in 400 mL of acetonitrile at room temperature, and the resulting mixture was stirred at reflux temperature for 16 hours. 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] + .

[0624] Preparation of 17-4

[0625] Under an inert atmosphere, to a solution of oxalyl chloride (572 mg, 4.51 mmol, 1.5 equiv) in anhydrous tetrahydrofuran (5 mL) was added dropwise a solution of dimethyl sulfoxide (704 mg, 9.02 mmol, 3.0 equiv) in tetrahydrofuran (3 mL) with stirring at -80 °C. After 30 minutes, ((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 hours, triethylamine (1.9 mL, 13.5 mmol, 4.5 equiv) was added and stirring was continued at -80 °C for 30 minutes. The mixture was warmed to room temperature and diluted with dichloromethane (20 mL). The organic layer was washed with 2×20 mL saturated ammonium chloride solution and 20 mL brine in sequence. 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.

[0626] Preparation of 17-5

[0627] At 0 ° C, sodium hydride (7.4 g, 309.7 mmol, 4.0 equivalents) was added to a solution of 17-3 (50.0 g, 77.43 mmol) in 500 mL of tetrahydrofuran and the resulting mixture was stirred at 0 ° C for 30 minutes. Then 17-4 (10.2 g, 61.9 mmol, 0.8 equivalents) was added at 0 ° C, and the mixture was stirred at room temperature for 16 hours. The reactant was quenched by adding a saturated ammonium chloride solution (1000 mL) and extracted with 3 × 1500 mL of dichloromethane. The combined organic layer was 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 obtain 31.0 g (88% yield) of 17-5 as a yellow oil. 1 H-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).

[0628] Preparation of 17-6

[0629] Palladium / carbon (25 g) was added to a solution of 17-5 (12.5 g, 27.7 mmol) in 250 mL of ethyl acetate, and the resulting mixture was stirred for 8 hours at 20 atm pressure and 65 ° C under hydrogen. The mixture was then filtered, the filtrate was concentrated, and the residue was purified on a silica gel column with 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).

[0630] Preparation of 17-7

[0631] To a solution of 17-6 (3.5 g, 9.7 mmol) in 70 mL of acetone at 0° C., Jones reagent (11.2 mL) was added, and the resulting mixture was stirred at 40° C. for 24 hours. 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).

[0632] Preparation of 17

[0633] To a solution of 17-7 (3 g, 8 mmol) in DMF (60 mL) at 0°C were 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). The resulting mixture was stirred at room temperature for 12 hours and then extracted with 3×500 mL of dichloromethane. The combined organic layers were washed with 2×500 mL of water and 500 mL of saturated brine. The organic layer was dried (Na2SO4), 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).

[0634] Embodiment 18 .

[0635]

[0636] Preparation of 18-1

[0637] At 0 ° C, under an argon inert atmosphere, heptadecanoic acid (5.8 g, 21.6 mmol, 1.0 equivalent) was added to a solution of trans-4-aminocyclohexanecarboxylic acid methyl ester hydrochloride (5 g, 25.9 mmol, 1.2 equivalents) and N, N-diisopropylethylamine (6.68 g, 51.8 mmol, 2.4 equivalents) in 90 mL of dichloromethane, and the mixture was stirred for 10 minutes. Then N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (4.97 g, 25.9 mmol, 1.2 equivalents) and 1-hydroxybenzotriazole (3.5 g, 25.9 mmol, 1.2 equivalents) were 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 with 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).

[0638] Preparation of 18-2

[0639] At 0 ° C, lithium hydroxide (5M, 43.4 mL, 87 mmol, 5.0 equivalents) was added to a solution of 18-1 (7.1 g, 17.4 mmol) in 71 mL of tetrahydrofuran under an inert atmosphere of argon. The resulting solution was stirred at room temperature for 16 hours and then neutralized to pH 6 with hydrochloric acid (2M). 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 with 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] + . 1HNMR(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).

[0640] Preparation of 18

[0641] At 0 ° C, to a solution of 18-2 (5 g, 12.6 mmol) and 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) under an inert atmosphere of nitrogen, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (3.63 g, 18.9 mmol, 1.5 equiv) was added. The resulting solution was stirred at room temperature for 16 hours 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 with hexane / ethyl acetate (70 / 1 to 1 / 1). The fractions containing the product 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).

[0642] Embodiment 19 .

[0643]

[0644] Preparation of 19-1

[0645] 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 to a solution of (-)-borneol (2 g, 12.7 mmol) in dichloromethane (30 mL) 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 with 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] + .

[0646] Preparation of 19-2

[0647] To a solution of 19-1 (6.7 g, 13.06 mmol) in tetrahydrofuran (67 mL) was added 10% palladium / activated carbon (1.4 g) and flushed with H2 for five cycles. The resulting mixture was stirred for 16 hours, then filtered, the solid was washed with tetrahydrofuran, and the combined filtrate was concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (50 / 1 to 10 / 1) to give 4 g (75% yield) of 19-2 as a white solid. MS: m / z 445.40 [M+Na] + .

[0648] Preparation of 19

[0649] 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 to a solution of 19-2 (5.5 g, 13.0 mmol) in N,N-dimethylformamide (55 mL) and stirred for 16 hours. 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 with 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] + . 1H-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).

[0650] Embodiment 20 .

[0651]

[0652] Preparation of 20-2

[0653] At 0 ° C, sodium hydride (2.6 g, 109.1 mmol, 2.0 equiv) was added 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) and stirred for 30 minutes, then 20-1 (25 g, 54.5 mmol, 1.0 equiv) was added at 0 ° C and the solution was stirred at 40 ° C for 16 hours. The reactant was quenched with saturated NH4Cl and extracted with dichloromethane. The combined organic layers were 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.

[0654] Preparation of 20-3

[0655] To a solution of 20-2 (2.3 g, 4.74 mmol) in THF (46 mL) was added palladium on carbon (4.6 g), and the mixture was stirred at 65°C for 8 hours under H2 (20 atm pressure). The resulting mixture was filtered, and the filtrate was concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 6 / 1) to give 1.8 g (96% yield) of 20-3 as a white solid. 1 HNMR(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).

[0656] Preparation of 20-4

[0657] To a solution of 20-3 (3.46 g, 8.8 mmol) in acetone (121 mL) was added Jones reagent (10.5 mL) at 0° C. and stirred at 40° C. for 12 hours. The reaction mixture was then filtered, concentrated and purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 6 / 1) to give 2.87 g (80% yield) of 20-4 as a white solid. 1 HNMR(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).

[0658] Preparation of 20

[0659] 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 hours. The resulting mixture was extracted with dichloromethane. The organic layer was washed with water and brine, dried (anhydrous Na2SO4), 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] + . 1HNMR(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).

[0660] Example 21.

[0661]

[0662] Preparation of 22-1

[0663] A solution of 16-(benzyloxy)-16-oxohexadecanoic acid (4.8 g, 12.8 mmol) in sulfuryl chloride (40 mL) was stirred at 70 ° C for one hour. It was then cooled to room temperature and concentrated. At room temperature, 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 equivalent) in tetrahydrofuran (10 mL) and stirred for 2 hours. The pH was adjusted to 7 with 1M 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 obtain 5 g (78% yield) of 22-1 as a colorless oil. MS: m / z 537.50 [M+Na] + .

[0664] Preparation of 22-2

[0665] To a solution of 22-1 (5 g, 7.41 mmol) in tetrahydrofuran (50 mL) was added 10% palladium / activated carbon (1 g) under N2. The flask was flushed with H2 for five cycles and stirred under H2 for 16 hours. The reaction mixture was then filtered, concentrated, and the residue was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 0 to 40 / 1) to give 3.5 g (74% yield) of 22-2 as a white solid. MS: m / z 447.40 [M+Na] + .

[0666] Preparation of 22

[0667] 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 to a solution of 22-2 (5.5 g, 11.656 mmol) in N,N-dimethylformamide (55 mL) and stirred for 16 hours. The reaction mixture was then diluted with ethyl acetate and the organic layer was washed with water and brine. The resulting solution was then dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified on a silica gel column with 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).

[0668] Embodiment 22 .

[0669]

[0670] Preparation of 23-1

[0671] 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 equivalents). The mixture was warmed to reflux and stirred for 48 hours. The resulting mixture was cooled to room temperature, filtered and concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (60 / 1 to 30 / 1) to give 35 g (47% yield) of 23-1 as a white solid.

[0672] Preparation of 23-2

[0673] At 0 ° C, sodium hydride (60% dispersion in oil, 8.75g, 218.8mmol, 2.0 equivalents) was added to a solution of 23-1 (35g, 109.4mmol) in anhydrous N, N-dimethylformamide (700mL) under Ar and stirred for 30 minutes. At 0 ° C, benzyl bromide (28.06g, 164.1mmol, 1.5 equivalents) was added to the reaction mixture and stirred at room temperature for 16 hours. 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 with petroleum ether / ethyl acetate (500 / 1 to 200 / 1) to obtain 30g (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).

[0674] Preparation of 20-1

[0675] Sodium iodide (28.125 g, 187.5 mmol, 2.0 equivalents) was added to a mixture of 23-2 (30 g, 93.75 mmol, 1.0 equivalents) in acetone (600 mL), and refluxed for 2 hours under vigorous stirring. The reaction mixture was cooled, filtered and concentrated. The residue was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 60 / 1) to obtain 30 g (89.5% yield) of 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).

[0676] Preparation of 23-3

[0677] Sodium hydride (60% dispersion in oil, 5.24 g, 131.0 mmol, 2.0 equiv) was added to a solution of L(-)-menthol (20.44 g, 131.0 mmol, 2.0 equiv) in anhydrous N,N-dimethylformamide (409 mL) at 0°C under Ar and stirred for 30 minutes. 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 hours. 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 with petroleum ether / ethyl acetate (200 / 1–100 / 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).

[0678] Preparation of 23-4

[0679] To a solution of 23-3 (7.2 g, 14.81 mmol, 1.0 equiv) in ethyl acetate (144 mL) was added palladium / activated carbon (10%, 14.4 g) and acetic acid (0.89 g, 14.8 mmol, 1.0 equiv) under H2 and stirred at 65°C for 8 hours. The solution was filtered, concentrated, and the residue was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 50 / 1) to give 5.2 g (88.6% yield) of 23-4 as a white solid. 1 H 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).

[0680] Preparation of 23-5

[0681] To a solution of 23-4 (5.2 g, 13.13 mmol, 1.0 equiv) in acetone (182 mL) at 0 °C under Ar was added Jones reagent (2.67 M, 15.74 mL, 42.02 mmol, 3.2 equiv) and stirred at 40 °C for 12 hours. The reaction mixture was filtered and concentrated, and the residue was diluted with dichloromethane, then 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 (40 / 1 to 8 / 1) to give 3.5 g (65% yield) of 23-5 as an aqueous solid. MS: m / z 409.30 [MH] - . 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).

[0682] Preparation of 23

[0683] At 0 ° C, 4-dimethylaminopyridine (0.21 g, 1.71 mmol, 0.2 equiv) and N,N-diisopropylethylamine (3.3 g, 25.61 mmol, 3.0 equiv) were added 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) under Ar and stirred for 30 minutes. 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 hours. The reaction mixture was 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 with 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] + . 1H 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).

[0684] Example 23.

[0685]

[0686] Preparation of 24-1

[0687] At room temperature, hexadecane-1-thiol (6.38 g, 24.69 mmol, 1.5 equivalents) was added to a solution of 1,1'-carbonyldiimidazole (4.8 g, 19.6 mmol, 1.8 equivalents) in 50 mL of dichloromethane under an inert atmosphere of argon. 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 filter 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 equivalents) and 1-hydroxybenzotriazole (3.33 g, 24.7 mmol, 1.5 equivalents) 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 Prep-Flash using the following conditions (C18-column, mobile phase, water and tetrahydrofuran (30% to 100% THF gradient)). The fractions containing the product were concentrated to give 6.9 g (80% yield) of 24-1 as a white solid. MS: m / z 528.15 [M+H] + . 1H 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).

[0688] Preparation of 24-2

[0689] 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 hours. The reactant 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 layer was dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by Prep-Flash using the following conditions (C18-column, mobile phase, water and THF, gradient 30% to 100% THF). The fractions containing the product 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 Na2SO4), filtered and concentrated to give 3.9 g (62% yield) of 24-2 as a white solid. MS: m / z 852.45 [M+Na] + . 1H 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).

[0690] Preparation of 24

[0691] Under an argon inert atmosphere, 3-(bis(diisopropylamino)phosphinoyl)propionitrile (1.35 g, 4.48 mmol, 1.2 equiv.), 4,5-dicyanoimidazole (485 mg, 4.1 mmol, 1.1 equiv.) were added to a solution of 24-2 (3.1 g, 3.74 mmol) in dichloromethane (31 mL), and the reaction mixture was stirred at room temperature for 2 hours. The resulting solution was diluted with 500 mL of dichloromethane, washed with 300 mL of saturated aqueous sodium bicarbonate solution and 300 mL of saturated brine. The organic layer was dried (anhydrous Na2SO4), filtered and concentrated. The crude product was purified by Prep-Flash using the following conditions (C18-column, mobile phase, water and THF, gradient 30% to 100% THF). The fractions containing the product were combined and diluted with an equal volume of dichloromethane. The organic layer was separated. The aqueous phase was extracted with 3×200 mL of dichloromethane. The combined organic extracts were dried (anhydrous Na2SO4), filtered and concentrated. The crude residue was purified on a silica gel column with 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] + . 1H 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)。 31 P NMR(DMSO-d6)δ149.13,147.57。

[0692] Embodiment 24 .

[0693]

[0694] Preparation of 25-1

[0695] At ℃, 4-methylbenzenesulfonyl chloride (8.1 g, 42.5 mmol, 1.1 equiv) was added 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. The resulting solution was stirred at room temperature for 2 hours. The reactants were quenched by adding 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 Na2SO4, 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 for 2 hours under vigorous stirring, 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 Na 2 SO 4 , filtered and concentrated. The crude product was purified on a silica gel column with 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 = 7.0 Hz, 2H), 1.83 (t, J = 3.2 Hz, 3H), 1.71 (q, J = 7.1 Hz, 2H), 1.63-1.40 (m, 6H), 1.38-1.05 (m, 30H), 0.93 (d, J = 7.2 Hz, 2H).

[0696] Preparation of 25-3

[0697] At 0 ° C, sodium hydride (0.71 g, 29.6 mmol, 4.0 equiv.) was added to a solution of 25-2 (3.1 g, 7.4 mmol) in 70 mL of tetrahydrofuran and the resulting mixture was stirred at 0 ° C for 30 minutes. Then 25-1 (13.9 g, 29.6 mmol, 4.0 equiv.) was added at 0 ° C and the reaction mixture was stirred at 65 ° C for 6 hours. The reactants were quenched by adding 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 Na2SO4, filtered and concentrated. The crude product was purified on a silica gel column with 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] + .

[0698] Preparation of 25-4

[0699] To 25-3 (3.7 g, 4.811 mmol, 1.0 equivalent) in a solution of methanol (37 mL) and tetrahydrofuran (37 mL) was added palladium hydroxide (3.7 g) and stirred for 2 hours under H2 atmosphere. The mixture was filtered and the filtrate was concentrated. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 3 / 1) to obtain 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).

[0700] Preparation of 25-5

[0701] 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 hours. The resulting solution was extracted with 3×100 mL of dichloromethane. The combined extracts were washed with 2×100 mL of water and 100 mL of saturated brine, dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified on a silica gel column with petroleum ether / ethyl acetate (100 / 1 to 1 / 2) to give 1.3 g (85% yield) of 25-5 as a white solid. 1H-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).

[0702] Preparation of 25

[0703] 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) were added to a solution of 25-5 (3.3 g, 3.7 mmol) in dichloromethane (33 mL). The resulting solution was stirred at room temperature for 2 hours and then extracted 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 with 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] + . 1 H-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.

[0704] Embodiment 25 .

[0705] The following compounds were prepared in a similar manner to the compounds provided in the preceding examples using the corresponding starting materials.

[0706]

[0707]

[0708]

[0709]

[0710] Example 26. Knockdown efficiency of MAPT siRNA in human iPSC neurons .

[0711] MAPT siRNA agents conjugated to certain lipophilic monomers provided herein were synthesized and the sequences are listed in Table 3.

[0712] Table 3: Modified sense and antisense strand sequences (5' to 3') of MAPT siRNA conjugated to different lipids

[0713] Modifications: mN = 2'OMe; fN = 2'F; ps = phosphorothioate; VP = vinylphosphonate; invAb = inverse desorption Base

[0714] Chol4 = cholesterol-triethylene glycol

[0715]

[0716]

[0717]

[0718]

[0719]

[0720]

[0721]

[0722] The in vitro knockdown efficiency of lipid-siRNA conjugates was evaluated in human iPSC-derived cortical neurons following the procedure described below.

[0723] Differentiation of human iPSCs into cortical neurons

[0724] The human iPSC line (Sigma # iPSC0028) was obtained by reprogramming epithelial cells from a 24-year-old white female donor via OSKM retroviral reprogramming. iPSCs were first differentiated into cortical neural stem cells (NSCs) according to a modified dual SMAD inhibitor protocol (Shi et al., 2012, 2012, Nat. Proc. 7 (10): 1836-46). In brief, iPSCs were plated at 500,000 cells / cm 2The cells were plated on wells coated with Matrigel (Corning 354230) in mTeSR medium (StemCell Technologies 5850) supplemented with 10 μM ROCK inhibitor (Sigma-Aldrich Y0503) and cultured at 37° C. and 5% O 2 . The medium was replaced with mTeSR the next day (Day -1). From day 0 to day 12, the cell culture medium was replaced daily with cortical neural induction medium consisting of 10 μM SB43142 (Tocris 1614) and 1 μM Dorsomorphin (Tocris 3093) supplemented with neural maintenance medium (1:1 DMEM:F12 Glutamax (ThermoFisher Scientific 10565108), Neurobasal (ThermoFisherScientific21103049), 2.5 μg / mL insulin (Sigma-Aldrich I9278-5ML), 50 μM 2-mercaptoethanol (ThermoFisher Scientific, 31350010), 0.5% non-essential amino acids (ThermoFisher Scientific11140035), 0.5% GlutaMAX supplement (ThermoFisher Scientific, 10565018), 0.5 mM sodium pyruvate (ThermoFisher Scientific, 10565019). Scientific 11360070), 1% penicillin-streptomycin (Sigma P4333), 0.5% N2 supplement (ThermoFisher Scientific 17502048), 1% B27 supplement (ThermoFisher Scientific17504044). On day 12, the neuroepithelial sheets were gently dissociated into large clumps of 300 to 500 cells using a needle and lifter, and the clumps were collected in a 15 mL falcon by centrifugation at 160 g for 2 minutes. The cell pellet was gently resuspended in neural induction medium and passaged 1 / 2 or 1 / 3 into wells coated with 10 μg / mL laminin (Sigma-Aldrich L2020), with a total volume of 2 mL neural induction medium per well in a 6-well plate. On days 13 and 15, the medium was changed to medium supplemented with 20 ng / mL FGF2 (Stemcell Technologies On day 17, neural rosettes were dissociated using dispase (ThermoFisher Scientific 17105041), passaged 1 / 3, and seeded into laminin-coated wells.1 or 2 additional dispersion steps were performed for one week. At approximately day 25-30, neural stem cells were dissociated into single cell suspensions using 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. Cryovials of neural stem cells were stored in liquid nitrogen until use.

[0725] To generate iPSC neurons, cryopreserved vials of neural stem cells (NSCs) were thawed and plated at 70,000 cells / cm 2 Seed in laminin coated wells, the wells are equipped with neural maintenance medium supplemented with 10 μM ROCK inhibitor and 20ng / mL FGF2. In the next two days, the medium is replaced with neural maintenance medium every day. About the 4th day after thawing, the cells are dissociated with Accutase and placed in a 96-well plate pre-coated with poly-L-ornithine and laminin at 28,000 cells / well in neural maintenance medium supplemented with 10 μM ROCK inhibitor. The next day after re-plating, the medium is replaced with neural differentiation medium, which is composed of neural maintenance medium supplemented with 20ng / mL BDNF (R&D Systems 212-BD-050 / CF), 20ng / mL GDNF (R&D Systems 212-GD-050 / CF), 500μM DB-cAMP (Sigma D0627) and 20mM ascorbic acid (Sigma A4403). Cultures were differentiated in neural differentiation medium with 50% medium changes twice a week. After two to three weeks of differentiation from neural stem cells (NSCs), neurons were treated with siRNA for 7 or 14 days and analyzed for RNA by reverse transcription and real-time PCR and for protein by MSD immunoassay.

[0726] MSD immunoassay

[0727] Human iPSC neurons differentiated on a 96-well plate were lysed in 100 μL of ice-cold RIPA buffer (Sigma) supplemented with cOmpleteTM protease inhibitor cocktail (Roche) and PhosSTOPTM (Roche) and gently orbitally shaken for 30 minutes at 4°C. The plate was centrifuged at 1,000 × g for 5 minutes, and the cell lysate was collected and diluted at different (20, 50 or 100) times, and then protein measurements were performed using the MSD immunoassay according to standard procedures. Briefly, the MSD plate was coated with 30 μL of coating antibody diluted in PBS at 1 μg / mL per well and placed at 4°C overnight. The next day, the plate was inverted on absorbent paper to dry, and then incubated with 150 μL of 0.1% casein blocking buffer per well for 2 hours at room temperature while orbitally shaking at 300 rpm. After blocking, the plates were incubated overnight at 4 ° C with cell lysates diluted in RIPA buffer supplemented with cOmpleteTM protease inhibitor cocktail (Roche). The plates were washed 5x times using Titertek Aquamax 4000 and inverted on absorbent paper to dry. Then, 25 μL / secondary antibody 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 5x times and developed with 150 μL / well 2x MSD reading buffer (Meso Scale Discovery) diluted in milliQ H2O. The plates were read immediately using an MSD instrument. For the measurement of Tau protein, hTau43 antibody (Janssen) was used for 96-well MSD plate coating, and SULFO-TAGTM 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 14221BF (Cell Signaling Technology) was used as primary detection, and SULFP-TAG-labeled anti-mouse antibody (Meso Scale Discovery) was used as secondary detection antibody.

[0728] Tables 4 to 6 summarize the efficiency of MAPT siRNA conjugated to different lipids in knocking down MAPT mRNA in human iPSC neurons after 7 days of incubation. Three concentrations of siRNA conjugates of 1 μM, 200 nM and 40 nM were tested. Data are expressed as the remaining MAPT mRNA relative to the control. Mean ± SD, n = 3.

[0729] Table 4. MAPT mRNA knockdown efficiency of MAPT siRNA M28 lipid conjugates in human iPSC neurons .

[0730]

[0731] Table 5. MAPT mRNA knockdown efficiency of MAPT siRNA M25 lipid conjugates in human iPSC neurons .

[0732]

[0733]

[0734] Table 6. MAPT mRNA knockdown efficiency of MAPT siRNA M25 lipid conjugate (lipid walking) in human iPSC neurons Rate .

[0735]

[0736] Example 27. Stability study

[0737] The in vitro stability of siRNA conjugates in various matrices including mouse brain homogenate, human liver lysosomes, and rat liver trisomes was assessed using the following method. The results of the analysis are provided in Tables 7 and 8.

[0738] Tables 7 and 8 summarize the LC-MS measurements of antisense and sense strand stability of siRNA conjugates in mouse brain homogenates (Tables 7 and 10) and human liver lysosomes (Tables 8 and 11) and rat liver trisomes (Table 12) after incubation at 37°C with shaking at 450 rpm for 24 hours. The reference compound MSC-2-V was added to the assay to benchmark the stringency of the assay conditions.

[0739] Table 7: LC-MS measurement of MAPT siRNA stability in mouse brain homogenate

[0740]

[0741] Table 8: LC-MS measurement of MAPT siRNA stability in human liver trisomy

[0742]

[0743] Table 9: Lipid-M28Var conjugates in hTAU KI mice 7 days after ICV injection (15 nmol siRNA conjugate) In vivo knockdown efficiency. Data are presented as residual MAPT mRNA (%) levels; n = 6 mice per treatment group .

[0744]

[0745]

[0746] Table 10: LC-MS measurement of MAPT siRNA stability in mouse brain homogenate .

[0747]

[0748] Table 11: LC-MS measurement of MAPT siRNA stability in human liver trisomy .

[0749]

[0750] Table 12: LC-MS measurement of MAPT siRNA stability in rat liver trisomy .

[0751]

[0752] Example 28: Evaluation of in vivo knockdown efficiency of siRNA conjugates in mouse models

[0753] The in vivo knockdown efficiency of lipid-siRNA conjugates was evaluated by intracerebroventricular (ICV) injection in a mouse model according to the following procedure.

[0754] After 7 days of single ICV injection of M28_Var1, M28_Var39, M28_Var40, M28_Var41, M28_Var42, M28_Var50 at 15 nmol, MAPT mRNA in seven brain regions (cortex, hippocampus, brainstem, cerebellum, striatum, midbrain and cervical spinal cord) from hTAU KI mice was evaluated. The results are in Table 9. Data are expressed as residual MAPT mRNA (%). Mean ± SD, n = 6 mice / treatment group.

[0755] Mice and intracerebroventricular injection

[0756] Male and female PS19 mice (C57BL6; Prnp-MAPT*P301S) or hTau KI (C57BL6; hMAPT: knock-in) mice aged 2 to 3 months were randomly assigned to different treatment groups. Mice were anesthetized with isoflurane (induction: 4%-5%; maintenance: 1.8%-2.5%). They were then stereotaxically injected into the bilateral ventricles using an electric drill and a microinjection robot (Neurostar, Germany, SterodriveSofeware v2019) with coordinates: AP: -0.62mm, ML: + / -1.05mm, and DV: 2.2mm. The volume of each injection was 5 μL for 5 minutes, and the needle was removed in three steps after injection (1. 1mm was removed within 60 seconds and maintained for 5 minutes; 2. 0.5mm was removed within 30 seconds and waited for 5 minutes; 3. It was removed from the brain at a very slow speed) to avoid the compound from flowing back along the needle track. After each step, the amount of reflux was checked. On selected days after injection, animals were sacrificed and different regions of the brain, including the cortex, hippocampus, brainstem, cerebellum, striatum, midbrain and cervical spinal cord, were dissected and then snap-frozen and stored at -80°C until further analysis.

[0757] RNA extraction from tissue

[0758] The tissues were collected in 2 mL tubes containing 1.4 mm ceramic beads Lysing Matrix D (MP Biomedicals 6913-500) and stored in a -80°C freezer until analysis. The tissues were placed on ice under laminar flow and 750 μL Trizol (ThermoFisher 15596026 / 15596018) was immediately added to each tube. FastPrep-24 TM 5G grinder, 3 cycles of 30 seconds at a speed of 5m / s, the tissue is broken and homogenized. Between cycles, the sample is cooled on ice for 2 minutes. After homogenization, the test tube is immediately rotated and 20% volume of chloroform is added (when using 750μL Trizol, 150μL chloroform is added). Vortex 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, and 1 volume of 70% ethanol is added and mixed thoroughly. The next steps are performed using the RNeasy 96 kit (Qiagen) according to the manufacturer's protocol. The RNeasy96 plate is placed on top of the Square-Well Block support, and the sample (Trizol / chloroform extraction) is placed in the well of the RNeasy 96 plate and the plate is sealed with an AirPore cover to prevent contamination. Centrifuge at room temperature for 3 minutes at 5600×g, and then the solution is discarded. The successive washing steps included applying 1 x 800 μL RW1 buffer, 2 x 800 μL RPE buffer to the RNeasy 96 plate, and removing the wash buffer by centrifugation at 5600 × g for 3 minutes for each step. After the last wash, the RNeasy 96 plate was centrifuged at 5600 × g for 3 minutes to remove residual lipids. RNA was eluted using 60 μL RNase-free water at 5600 × g for 3 minutes at room temperature. RNA concentration was measured by Nanodrop 8000 (ThermoFisher). The eluted RNA was stored at -80 °C until analysis.

[0759] RT-qPCR

[0760] RNA was used as a template for reverse transcription, and reverse transcription was performed using a high-capacity cDNA reverse transcription kit (Applied Biosystems) according to the manufacturer's protocol. Briefly, the final reaction of 20 μL of the mixture 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 qPCR reactions, the reverse transcribed cDNA was diluted 10-fold and mixed with 2X PowerUp TM SYBR TMGreen Mater Mix (ThermoFisher A25743) and 500 nM qPCR primers were mixed to a final reaction volume of 10 μL. TM 12K instrument (Applied Biosystems TM ) qPCR runs were performed using a standard thermal cycling protocol. Multiple primers were used to detect MAPT mRNA, and reference primers targeting mouse housekeeping genes AP3D1 and PAK1IP1 were included to normalize gene expression.

[0761] The above embodiments are intended to be exemplary only, and those skilled in the art will recognize or be able to ascertain, using only routine experimentation, many equivalents to the specific compounds, materials, and procedures. All such equivalents are considered to be within the scope of the invention and are covered by the appended claims.

Claims

1. A compound of formula (I): in: X is O or S; Y is O or NR a ; 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 to which they are attached and for Y; Z 1 , Z 2 and Z 3 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 * refers to the direction toward the ring containing X; R 1 , R 2 and R 3 Each is independently H, C1-C6 alkyl optionally substituted by C1-C6 alkoxy, or -(GL) m -R, condition is R 1 , R 2 and R 3 At least one of -(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 -; where * refers to the direction toward 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; and Each instance of R is independently C 12 -C 32 Alkyl or C 12 -C 32 alkenyl; x is 1 or 2; n is 0 or 1; m is 0, 1 or 2; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

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

3. The compound according to claim 1, wherein X is S.

4. The compound according to any one of claims 1 to 3, wherein Y is O.

5. The compound according to any one of claims 1 to 3, wherein Y is NH.

6. A compound according to any one of claims 1 to 5, wherein Z 1 , Z 2 and Z 3 Each is independently -O-, -NH-, *-OC(=O)-, *-NHC(=O)-, -OC(=O)O-, *-NHC(=O)O-, *-OC(=O)NH- or -NHC(=O)NH-.

7. The compound according to claim 6, wherein Z 1 , Z 2 and Z 3 All are -O-.

8. A compound according to any one of claims 1 to 7, which is a compound of formula (IA): or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

9. The compound according to claim 8, which is a compound of formula (IA-1): or a pharmaceutically acceptable salt thereof.

10. The compound according to claim 8, which is a compound of formula (II-A): or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

11. The compound according to claim 10, which is a compound of formula (II-A-1): or a pharmaceutically acceptable salt thereof.

12. A compound according to any one of claims 8 to 11, wherein R 1 -(GL) m -R, and R 2 It is a C1-C6 alkyl group which is optionally substituted by a C1-C6 alkoxy group.

13. A compound according to any one of claims 8 to 11, wherein R 2 -(GL) m -R, and R 1 It is a C1-C6 alkyl group which is optionally substituted by a C1-C6 alkoxy group.

14. A compound according to any one of claims 8 to 11, wherein R 1 and R 2 Each independently is -(GL) m -R.

15. A compound according to any one of claims 8 to 14, wherein -(GL) m -R is -R.

16. A compound according to any one of claims 8 to 14, wherein -(GL) m -R is (C2- C3 alkylene)-NH-C(=O)-R.

17. A compound according to any one of claims 1 to 7, which is a compound of formula (IB): or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

18. The compound according to claim 17, which is a compound of formula (IB-1): or a pharmaceutically acceptable salt thereof.

19. The compound according to claim 17, which is a compound of formula (II-B): or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

20. The compound according to claim 19, which is a compound of formula (II-B-1): or a pharmaceutically acceptable salt thereof.

21. A compound according to any one of claims 17 to 20, wherein R 1 and R 2 Each independently is -(GL) m -R, and R 3 It is a C1-C6 alkyl group which is optionally substituted by a C1-C6 alkoxy group.

22. A compound according to any one of claims 17 to 20, wherein R 1 and R 3 Each independently is -(GL) m -R, and R 2 It is a C1-C6 alkyl group which is optionally substituted by a C1-C6 alkoxy group.

23. A compound according to any one of claims 17 to 20, wherein R 2 and R 3 Each independently is -(GL) m -R, and R 1 It is a C1-C6 alkyl group which is optionally substituted by a C1-C6 alkoxy group.

24. A compound according to any one of claims 17 to 23, wherein -(GL) m -R is -R.

25. A compound according to any one of claims 17 to 23, wherein -(GL) m -R is -(C2-C3 alkylene)-NH-C(=O)-R.

26. A compound of formula (III): in: B is a modified or unmodified nucleobase; X is O or S; Y is O or NR a ; 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 to which they are attached and for Y; 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 a C(=S)NR b -、*-OC(=O)S-、*-NR a C(=O)S-、*-OS(O) x -or*-NR a S(O) x -; where * refers to the direction toward the ring containing X; R 2 -(GL) m -R; Each instance of G is independently C1-C8 alkylene or C3-C 10 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 -; where * refers to the direction toward 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; and R is C 12 -C 32 Alkyl or C 12 -C 32 alkenyl; x is 1 or 2; and m is 0, 1 or 2; The condition is when X is O, Y is O and Z 2 When it 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-、*-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 * refers to the direction towards R; and R" is a monocyclic, fused, bridged or spirocyclic 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 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 C3-C 10 Cycloalkylene; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

27. The compound according to claim 26, wherein X is O.

28. The compound of claim 26, wherein X is S.

29. A compound according to any one of claims 26 to 28, wherein Y is O.

30. A compound according to any one of claims 26 to 28, wherein Y is NH.

31. A compound according to any one of claims 26 to 30, which is a compound of formula (III-A): or a pharmaceutically acceptable salt thereof.

32. The compound according to claim 31, which is a compound of formula (III-A-1): or a pharmaceutically acceptable salt thereof.

33. A compound according to any one of claims 26 to 32, wherein Z 2 It is -OC(=O)NH-, -NHC(=O)-, -NHC(=O)NH- or -NHC(=O)S-.

34. according to the compound described in any one of claims 26 to 32, wherein X is O, Y is O, and Z 2 is -O-, then m is 0, and R is -L"-(CH2) 0-3 -R" instead.

35. The compound of claim 34, wherein R is substituted with adamantyl.

36. The compound according to claim 34, wherein R is replace.

37. A compound according to any one of claims 26 to 32, wherein X is O, Y is O, and Z 2 If -O-, then R 2 It is -(C2-C6 alkylene)-C(=O)NH-R or -(C2-C6 alkylene)-OR.

38. The compound according to any one of claims 26 to 37, wherein B is adenine (A), guanine (G), thymine (T), cytosine (C) or uracil (U).

39. A compound according to any one of claims 1 to 38, wherein D 1 A hydroxyl protecting group.

40. The compound according to claim 39, wherein D 1 It is 4,4'-dimethoxytrityl chloride (DMTr).

41. A compound according to any one of claims 1 to 40, wherein D 2 is a reactive phosphorus group.

42. The compound according to claim 41, wherein D 2 for 43. A compound according to any one of claims 1 to 38, wherein D 1 H and D 2 For H.

44. A compound of formula (IV): in: Each instance of G' is independently C1-C8 alkylene or C3-C8 cycloalkylene; Each instance of L' is independently *-NR c C(=O)-; wherein * refers to the direction toward the pyrrolidine-2,5-dione ring; R c Each instance of is independently H, C1-C 32 Alkyl or C2-C 32 alkenyl; R is C4-C 32 Alkyl or C4-C 32 alkenyl; and m is 0, 1 or 2; or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

45. The compound of claim 44, wherein m is 0.

46. ​​The compound of claim 44, wherein m is 1.

47. The compound of claim 46, wherein -(G'-L') m -R is 48. The compound of claim 44, wherein m is 2.

49. The compound of claim 48, wherein -(G'-L') m -R is 50. A compound according to any one of claims 1 to 49, wherein R is C 14 -C 18 alkyl.

51. A compound according to any one of claims 1 to 50, wherein R is unsubstituted.

52. A compound according to any one of claims 1 to 50, wherein 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-、*-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 * refers to 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; and R" is a monocyclic, fused, bridged or spirocyclic 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.

53. The compound according to claim 52, wherein the ring portion is C3-C 10 Cycloalkyl.

54. The compound of claim 52, wherein the ring portion is a 4- to 10-membered heterocyclyl containing 1 to 4 heteroatoms independently selected from O, N, and S.

55. The compound according to claim 52, wherein R is replace.

56. A compound according to any one of claims 52 to 55, wherein R is substituted at a terminal position.

57. A compound of Table 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

58. An oligonucleotide comprising at least one lipophilic monomer of the formula: Where X, Y, Z 1 , Z 2 , Z 3 , R 1 , R 2 , R 3 , B, G', L', R, n and m are as defined in any one of claims 1 to 57.

59. The oligonucleotide of claim 58, wherein the oligonucleotide is an antisense, an antagonist, a microRNA, a siRNA, a pre-microRNA, an antimir, a ribozyme, an RNA activator, a U1 adaptor, an immunostimulator, or an aptamer.

60. The oligonucleotide of claim 59, wherein the oligonucleotide is a siRNA comprising: The antisense strand is complementary to the target gene; A sense strand is complementary to the antisense strand.

61. The oligonucleotide of claim 60, wherein the lipophilic monomer is present in the antisense strand or the sense strand.

62. The oligonucleotide of claim 60, wherein the lipophilic monomer is present in the sense strand.

63. The oligonucleotide of claim 60, wherein the lipophilic monomer is conjugated to the 3' end or the 5' end of the sense strand or the antisense strand.

64. The oligonucleotide of claim 60, wherein the lipophilic monomer is located at an internal position of the sense strand or the antisense strand.

65. The oligonucleotide of any one of claims 60 to 64, wherein the sense strand and the antisense strand are each 15 to 30 nucleotides in length.

66. The oligonucleotide of claim 65, wherein the sense strand and the antisense strand are each 15 to 25 nucleotides in length.

67. The oligonucleotide of claim 66, wherein the sense strand and the antisense strand are each 21 to 23 nucleotides in length.

68. The oligonucleotide of any one of claims 60 to 67, further comprising a targeting ligand.

69. An oligonucleotide comprising at least one lipophilic monomer in Table 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

70. A method of delivering an oligonucleotide to a cell, comprising administering to the cell an oligonucleotide according to any one of claims 58 to 69.

71. The method of claim 70, wherein said administering results in an increased amount of said oligonucleotide delivered to said cell compared to delivery of the same oligonucleotide but without said lipophilic monomer.

72. The method of claim 71, wherein the increased amount of the oligonucleotide delivered is determined by an increase in the therapeutic activity of the oligonucleotide.

73. A method of reducing the expression of a target gene in a cell, comprising contacting the cell with an oligonucleotide according to any one of claims 58 to 69.

74. A method of reducing expression of a target gene in a subject, comprising administering to the subject an oligonucleotide according to any one of claims 58 to 69.

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