Phosphatidylserine derivative with high neuroprotective activity and application of phosphatidylserine derivative in treatment of Alzheimer's disease and Parkinson's disease

Abstract

The invention relates to a phosphatidylserine derivative with high neuroprotective activity and application of the phosphatidylserine derivative in treatment of Alzheimer's disease and Parkinson's disease. Specifically, the present invention provides a compound of formula I, or a tautomer, a stereoisomer, or a pharmaceutically acceptable salt thereof, in which the definitions of each group and substituent are described in the specification and claims. The invention further provides a preparation method of the compound, and the novel phosphatidylserine derivative which has good nerve cell proliferation promoting activity, nerve injury protection activity and the like and can be used for preparing drugs for preventing and / or treating neurodegenerative diseases.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medical and chemical industry, and particularly relates to phosphatidylserine derivatives with high neuroprotective activity and application thereof in treating Alzheimer's disease and Parkinson's disease. BACKGROUND

[0002] Phosphatidylserine (PS) is a key acidic phospholipid with high content in the brain (accounting for 13-15% of cortical phospholipids), mainly distributed on the cytoplasmic side of the neural cell membrane, and is a core component of synaptic membrane structure. A large number of studies have shown that PS not only has a significant regulatory effect on learning and memory function, but also exhibits a clear neuroprotective effect in various neuroprotective disease models: it can reduce the toxicity of beta-amyloid and regulate the function of the cholinergic system in Alzheimer's disease (AD) models; it shows an antidepressant-like effect in the treatment of depression; and it can improve the memory disorder induced by reserpine in Parkinson's disease (PD) models. However, there are still important technical limitations in current research. The existing research results are mostly based on PS mixed components, and the structure-activity relationship of specific PS molecular subtypes is not systematically analyzed, which makes it difficult to optimize the structure of PS in a targeted manner, and seriously restricts the in-depth development of its therapeutic potential.

[0003]

[0004] Therefore, there is an urgent need in the art to develop a new type of PS derivative, reveal its structure-activity relationship, and have the potential to develop as a drug for the treatment of neurodegenerative diseases. SUMMARY

[0005] The purpose of the present application is to provide a series of novel phosphatidylserine derivatives, their preparation methods and applications. The application value of the compounds of the present application in preventing and / or treating neurodegenerative diseases is verified by biological experiments.

[0006] In a first aspect, the present application provides a compound represented by formula I, or a tautomer, stereoisomer thereof, or a pharmaceutically acceptable salt thereof,

[0007]

[0008] wherein,

[0009] R1 is C5-C29 alkyl or C5-C29 alkenyl containing 1-5 unsaturations or is a substituent selected from the group consisting of:

[0010]

[0011] R2 is C5-C29 alkyl or C5-C29 alkenyl containing 1-5 unsaturations or is a substituent selected from the group consisting of:

[0012]

[0013] R A C5-C29alkyl, C5-C29alkenyl containing from 1 to 5 unsaturations,

[0014] R B H or 1 to 2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0015] X1or X2is -O- or -NH-; and X1and X2are not simultaneously -NH-;

[0016] X3is -O- or -NH-;

[0017] R3is selected from the group consisting of H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0018] R4is H or

[0019] with the additional proviso that:

[0020] R1or R2is not

[0021] and the compound is not

[0022] In another preferred embodiment, the compound of formula I is a compound of formula II,

[0023]

[0024] wherein,

[0025] R1is C5-C29alkyl, C5-C29alkenyl containing from 1 to 5 unsaturations, or a substituent selected from the group consisting of:

[0026] R2is C5-C29alkyl, C5-C29alkenyl containing from 1 to 5 unsaturations, or a substituent selected from the group consisting of:

[0027]

[0028] R A C5-C29alkyl, C5-C29alkenyl containing from 1 to 5 unsaturations,

[0029] R B H or 1 to 2 substituents each independently selected from the group consisting of halogen, -NO2, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6haloalkoxy;

[0030] X1or X2is -O- or -NH-; and X1and X2are not simultaneously -NH-;

[0031] X3is -O- or -NH-.

[0032] In another preferred embodiment, the compound of formula I is a compound of formula II-A,

[0033]

[0034] wherein,

[0035] R1is C5-C29alkyl containing 1 to 5 unsaturations or is a substituent selected from the group consisting of

[0036]

[0037] R2is C5-C29alkyl or C5-C29alkyl containing 1 to 5 unsaturations or is a substituent selected from the group consisting of

[0038]

[0039] R A selected from the group consisting of C5-C29alkyl, C5-C29alkyl containing 1 to 5 unsaturations,

[0040]

[0041] R B H or 1 to 2 substituents each independently selected from the group consisting of halogen, -NO2, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6haloalkoxy;

[0042] X3is -O- or -NH-;

[0043] with the additional proviso that:

[0044] when R1is C5-C29alkyl containing 1 to 5 unsaturations, R2is not C5-C29alkyl or C5-C29alkyl containing 1 to 5 unsaturations.

[0045] In another preferred embodiment, the compound is a compound of formula II-B1 or II-B2,

[0046]

[0047] wherein

[0048] R1is C5-C29alkyl containing 1 to 5 degrees of unsaturation or is a substituent selected from the group consisting of:

[0049] R2is C5-C29alkyl or C5-C29alkenyl containing 1 to 5 degrees of unsaturation or is a substituent selected from the group consisting of:

[0050]

[0051] R A selected from the group consisting of C5-C29alkyl, C5-C29alkenyl containing 1 to 5 degrees of unsaturation,

[0052]

[0053] R B is H or 1 to 2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0054] X3is -O- or -NH-.

[0055] In another preferred embodiment, the compound is a compound of formula III,

[0056]

[0057] wherein

[0058] R1and R2are each independently C5-C29alkyl or C5-C29alkenyl containing 1 to 5 degrees of unsaturation;

[0059] R3is selected from the group consisting of H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0060] R4is H or

[0061] R B is H or 1 to 2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy.

[0062] In another preferred embodiment, R3is selected from the group consisting of H, halogen, C1-C6alkyl, C1-C6haloalkyl.

[0063] In another preferred embodiment, R3 is H or C1-C6 alkyl.

[0064] In another preferred embodiment, R1 is selected from the group consisting of:

[0065]

[0066] and / or

[0067] R2 is C5-C29 alkyl or C5-C29 alkenyl containing 1-5 unsaturations;

[0068] wherein, R A , R B and X3 are as defined above.

[0069] In another preferred embodiment, R1 is selected from the group consisting of:

[0070]

[0071] In another preferred embodiment, R1 is selected from the group consisting of:

[0072] In another preferred embodiment, R1 is

[0073] In another preferred embodiment, R1 is

[0074] In another preferred embodiment, R1 is

[0075] In another preferred embodiment, X3 is O.

[0076] In another preferred embodiment, R2 is C5-C20 alkyl.

[0077] In another preferred embodiment, R2 is C9-C17 alkyl.

[0078] In another preferred embodiment, R2 is C5-C29 alkenyl containing 1-5 unsaturations.

[0079] In another preferred embodiment, R2 is C5-C29 alkenyl containing 2-5 unsaturations.

[0080] In another preferred embodiment, R2 is C5-C20 alkenyl containing 2-5 unsaturations.

[0081] In another preferred embodiment, R2 is C10-C20 alkenyl containing 2-5 unsaturations.

[0082] In another preferred embodiment, R2 is C15-C19 alkenyl containing 2-5 unsaturations.

[0083] In another preferred embodiment, R2is

[0084] In another preferred embodiment, R B is halogen or -NO2.

[0085] In another preferred embodiment, R B is -NO2.

[0086] In another preferred embodiment, R1is C5-C29alkyl containing 1 to 5 unsaturations; and / or

[0087] R2is selected from the group consisting of:

[0088] wherein R A , R B and X3are as defined above.

[0089] In another preferred embodiment, R1is C5-C20alkyl.

[0090] In another preferred embodiment, R1is C9-C17alkyl.

[0091] In another preferred embodiment, R1is C5-C29alkyl containing 1 to 5 unsaturations.

[0092] In another preferred embodiment, R1is C5-C29alkyl containing 2 to 5 unsaturations.

[0093] In another preferred embodiment, R1is C5-C20alkyl containing 2 to 5 unsaturations.

[0094] In another preferred embodiment, R1is C10-C20alkyl containing 2 to 5 unsaturations.

[0095] In another preferred embodiment, R1is C15-C19alkyl containing 2 to 5 unsaturations.

[0096] In another preferred embodiment, R2is selected from the group consisting of:

[0097] In another preferred embodiment, R2is selected from the group consisting of:

[0098] In another preferred embodiment, R2is

[0099] In another preferred embodiment, R2is

[0100] In another preferred embodiment, R2is

[0101] In another preferred embodiment, R B is selected from the group consisting of H, halogen, -NO2, C1-C6haloalkyl, C1-C6alkoxy.

[0102] In another preferred embodiment, R B is -NO2.

[0103] In another preferred embodiment, the compound is selected from the group consisting of:

[0104]

[0105]

[0106] In a second aspect of the present application, a method for preparing the compound of the first aspect of the present application is provided, and the method for preparing the compound of II-A is as follows:

[0107]

[0108] (S1) esterification of the compound of formula 1a with fatty acids with different or same substituents of R1and R2in the presence of a base to obtain intermediates 3a-1~3a-30;

[0109] (S2) deprotection of intermediates 3a-1~3a-30 by TFA treatment in an inert solvent to obtain intermediates 4a-1~4a-30;

[0110] (S3) coupling of intermediates 4a-1~4a-30 with tert-butyl serine and tert-butyl tetraisopropyl phosphorodiamidite to form phosphite triester intermediates 5a-1~5a-30 in the presence of an inert solvent and nitrogen;

[0111] (S4) oxidation of intermediates 5a-1~5a-30 to obtain intermediates 6a-1~6a-30 in an inert solvent, and deprotection to prepare compounds A1~A30;

[0112] wherein each group is as described above.

[0113] In another preferred embodiment, the preparation method of B1, B3~B7 in the compound is as follows:

[0114]

[0115] (S1) esterification of the compound of formula 1b with fatty acids with different or same substituents of R1and R2in the presence of a base to obtain intermediates 3b-1~3b-6;

[0116] (S2) in an inert solvent, intermediates 3b-1 ~ 3b-6 are reacted with boron chloride to remove the protecting group, thereby obtaining intermediates 4b-1 ~ 4b-6;

[0117] (S3) in an inert solvent, intermediates 4b-1 ~ 4b-6 are subjected to condensation, oxidation and deprotection reactions to obtain compounds B1, B3 ~ B7.

[0118] wherein each group is as defined above.

[0119] In a third aspect, the present application provides a composition comprising:

[0120] (1) a compound according to the first aspect of the present application, or a tautomer, stereoisomer thereof, or a pharmaceutically acceptable salt thereof; and

[0121] (2) a pharmaceutically acceptable carrier.

[0122] In a fourth aspect, the present application provides use of a compound of formula I for the manufacture of a medicament for the prevention and / or treatment of a neurodegenerative disease.

[0123]

[0124] wherein,

[0125] R1is C5-C29alkyl or C5-C29alkenyl containing 1-5 unsaturations or is a substituent selected from the group consisting of:

[0126]

[0127]

[0128] R2is C5-C29alkyl or C5-C29alkenyl containing 1-5 unsaturations or is a substituent selected from the group consisting of:

[0129]

[0130] R A selected from the group consisting of C5-C29alkyl, C5-C29alkenyl containing 1-5 unsaturations,

[0131] R B is H or is 1-2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0132] X1or X2is -O- or -NH-; and X1and X2are not simultaneously -NH-;

[0133] X3 is -O- or -NH-;

[0134] R3 is selected from the group consisting of H, halogen, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6haloalkoxy;

[0135] R4 is H or

[0136] In another preferred embodiment, the disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, depression, Huntington's disease.

[0137] It should be understood that, within the scope of the present application, each of the technical features of the present application described above and each of the technical features specifically described hereinafter (e.g. in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they are not listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0138] Figure 1 The effect of phosphatidylserine derivatives on the body weight of AD mice is shown. Data are expressed as mean ± SD, n = 7 in each group, *P < 0.05, **P < 0.01, ***P < 0.001, differences are statistically significant compared with the blank group.

[0139] Figure 2 The phosphatidylserine derivatives improve the cognitive ability of AD mice. The number of times of crossing the platform in the spatial exploration test stage of mice is shown. (Data are expressed as mean ± SD, n = 7 in each group; data were analyzed by one-way ANOVA, *P < 0.05, **P < 0.01, ****P < 0.0001, differences are statistically significant compared with the Aβ group).

[0140] Figure 3 The number of cells in the hippocampus is shown. (Data are expressed as mean ± SD, n = 3 in each group; *P < 0.05 indicates that the data are statistically significant compared with the Aβ 1-42 group by one-way ANOVA).

[0141] Figure 4 The effect of phosphatidylserine derivatives on the activation of glial cells in the hippocampus of mice is shown. The figure shows the density of microglia in the hippocampus induced by Aβ 1–42 (data are expressed as mean ± SD, n = 3 in each group. Data were analyzed by one-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001, differences are statistically significant compared with the Aβ group).

[0142] Figure 5The phosphatidylserine derivative reduced the expression levels of inflammatory factors in the hippocampus of mice. Among them, (A) TNF-a, (B) IL-6, and (C) IL-1b mRNA expression levels. (Data were normalized to the mRNA expression of the Aβ 1–42 The mRNA expression of each group was normalized, and the average value ± SD was used, n = 3 in each group. The data were analyzed by one-way ANOVA, and *P < 0.05 indicated that the difference was statistically significant compared with the Aβ 1-42

[0143] Figure 6 The effect of derivative A24 on the expression of proteins CHRM5, p-PI3K, p-AKT, p-ERK, ERK, p-CREB, and SYN in the hippocampus of AD mice was shown. Among them, (A) The expression levels of CHRM5, p-PI3K, p-AKT, p-ERK, ERK, p-CREB, and SYN proteins in the hippocampus of mice in the blank group, the Aβ group, and the Aβ+A24 group were detected by WB. (B) Compared with the Aβ group, the average level of all detected proteins in the Aβ+A24 group was significantly increased. The data were expressed by the average value ± SD, n = 3 in each group. The data were analyzed by one-way ANOVA, *P < 0.05, **P < 0.01, ***P < 0.001, indicating that the difference was statistically significant compared with the Aβ group.

[0144] Figure 7 The behavior performance (falling latency) of mice on days 0, 7, 14, and 21 of PS administration in the rotarod test was shown. The data were calculated by one-way ANOVA test, expressed by the average value ± SD, n = 7 in each group.

[0145] Figure 8 The grip performance of mice in each group on day 20 of PS treatment in the grip test was shown (n = 7 in each group). The vertical axis unit was Newton (N). The data were calculated by one-way ANOVA test, expressed by the average value ± SD, n = 7 in each group.

[0146] Figure 9 The total distance of movement of PS treatment on day 19 in the open field test was shown. The data were calculated by one-way ANOVA test, expressed by the average value ± SD, n = 7 in each group. DETAILED DESCRIPTION

[0147] ​The present inventors have conducted extensive and in-depth research, based on the structural characteristics of PS, and have systematically modified and optimized three key structural units thereof: the polar head (serine amine / methylene modification), the glycerol skeleton (replacement of hydroxyl groups with amine groups), and the fatty acid tail (regulation of carbon chain length / saturation and introduction of functional groups), thereby systematically investigating the effects of structural parameters such as polarity, molecular charge distribution, and steric hindrance on biological activity, and a total of 49 novel PS derivatives have been designed and synthesized. On this basis, the present application has been completed.

[0148] Terms

[0149] In the present application, the terms used have the general meanings well known to those skilled in the art, unless otherwise specified.

[0150] In the present application, the term "halogen" refers to F, Cl, Br, or I.

[0151] In the present application, "C1-C6 alkyl" refers to a straight-chain or branched alkyl group comprising 1-6 carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, neopentyl, tert-pentyl, or the like. The analogous term "C1-C4 alkyl" has an analogous definition.

[0152] In the present application, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, and the like. The analogous term "C1-C4 alkoxy" has an analogous definition.

[0153] In the present application, the term "C2-C6 alkenyl" refers to a straight-chain or branched alkenyl group having 2 to 6 carbon atoms and containing at least one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl, and hexenyl, and the like.

[0154] In the present application, the term "C2-C6 alkynyl" refers to a straight-chain or branched alkynyl group having 2 to 6 carbon atoms and containing at least one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl, and hexynyl, and the like.

[0155] In the present application, the term "C3-C8 cycloalkyl" represents a cyclic aliphatic group consisting of 3 to 8 ring-forming carbon atoms, and the like; it is to be understood that the "cycloalkyl" groups described herein include not only monocyclic aliphatic groups, but also fused, spiro, and bridged ring systems consisting of multiple cyclic aliphatic groups; the "cycloalkyl" groups described herein include not only aliphatic groups in which the carbon atoms are fully saturated, but also aliphatic groups in which some of the carbon atoms have unsaturated bonds; examples of the "cycloalkyl" groups described herein include but are not limited to: etc. The "C3-C6 cycloalkyl" and similar terms have analogous definitions. The term "heteroalkyl" refers to an alkyl group as defined herein in which one or more carbon atoms in the chain are independently replaced with a heteroatom (N, O, or S). Examples of "heteroalkyl" groups include, but are not limited to, the following: -CH2OCH3, -CH2NH2, -CH2NHCH3, -CH2N(CH3)2, -CH2SCH3, etc. The "heteroalkyl" group as a substituent can be attached to the molecular backbone at any chemically-allowed position on the "heteroalkyl" group.

[0156] In the present application, the term "aryl" refers to monocyclic and bicyclic ring systems composed of a specified number of carbon atoms that obey the Hückel rule; it is to be understood that when the "aryl" group of the present application is a bicyclic ring system, it includes not only the case where all the rings are aromatic, but also the case where only one of the rings is aromatic and the other ring is a non-aromatic aliphatic ring.

[0157] In the present application, the term "C6-C10 aryl" refers to a cyclic ring system having 6 to 10 carbon atoms, and at least one of the rings is aromatic; examples of the "aryl" group of the present application include, but are not limited to etc. The "aryl" group as a substituent is attached to the molecular backbone at the aromatic ring.

[0158] In the present application, the term "heterocycloalkyl" refers to a cyclic group composed of a specified number of ring-forming atoms, containing at least one ring-forming heteroatom (N, O, or S), saturated or partially unsaturated, and non-aromatic; it is to be understood that the "heterocycloalkyl" group of the present application includes not only monocyclic heterocyclic ring systems, but also polycyclic heterocyclic ring systems such as fused, spiro, and bridged rings; when the "heterocycloalkyl" group is a polycyclic ring system, at least one of the rings contains a ring-forming heteroatom, and the other rings can contain a ring-forming heteroatom or can be a cycloalkyl group; for example, the term "4-8 membered heterocycloalkyl" refers to a monocyclic or polycyclic ring system having 4 to 8 ring-forming atoms, at least one of which is a heteroatom, saturated or partially unsaturated; the definitions of other similar terms are analogous; more preferably, the number of heteroatoms is 1 to 3. Examples include, but are not limited to, the following: etc. It is to be understood that the "heterocycloalkyl" group as a substituent can be attached to the molecular backbone at any chemically-allowed position on the "heterocycloalkyl" group.

[0159] In the present application, the term "heteroaryl" means a cyclic group with a specified number of ring-forming atoms, containing at least one ring-forming heteroatom (N, O or S), having aromaticity; unless otherwise specified, "heteroaryl" as used herein encompasses not only monocyclic heteroaromatic systems, but also polycyclic heteroaromatic systems, such as bicyclic heteroaryl, tricyclic heteroaryl, tetracyclic heteroaryl; when the "heteroaryl" is a polycyclic heteroaromatic system, at least one ring is aromatic, and the other rings can be aromatic or non-aromatic, and the heteroatoms can be in the aromatic rings or in the non-aromatic rings; polycyclic heteroaromatic systems include not only fused ring systems, but also bridged and spiro ring systems. The term "5-7 membered heteroaryl" means a cyclic group with 5 to 7 ring-forming atoms, at least one of which is a heteroatom, having aromaticity. The definitions of other similar terms are analogous.

[0160] In the present application, the term "halo" means substituted with halogen.

[0161] In the present application, the term "optionally" means that when there is a series of candidate groups to choose from, some of them can be selected, and some of them can not be selected.

[0162] In the present application, "each independently" means that when several substituents defined at the same time are selected from the same series of candidate groups, they are independent of each other, and they can be the same or different.

[0163] In the present application, the term "substituted" means that one or more hydrogen atoms on a specified group are replaced by a specified substituent. The specified substituent is the substituent described in the foregoing or the substituent appearing in each embodiment. Unless otherwise specified, a substituted group can have one substituent selected from a specified group at any substitutable position of the group, and the substituents at each position can be the same or different. Those skilled in the art will understand that the combinations of substituents contemplated by the present application are those stable or chemically achievable combinations.

[0164] In the present application, the term "1-6" means having 1, 2, 3, 4, 5 or 6, and each of the other similar terms has a similar independent meaning.

[0165] In the present application, the term "unsaturation" means a quantitative characterization of the relative content or number of unsaturated bonds in the molecular structure of an organic compound. It is used to describe the proportion of double bonds (C=C) and triple bonds (C≡C) between carbon atoms or cyclic structures in the molecular chain. The term "containing 1-5 unsaturations" means that the compound structure contains 1-5 unsaturated bonds, and other similar terms have similar meanings.

[0166] It is to be understood that when a group occurs more than one time in a compound, its definition in each occurrence is independent of its definition at every other occurrence. That is, the term "is selected from the group consisting of" has the same meaning as the term "is independently selected from the group consisting of".

[0167] Compounds of formula I and processes for their preparation

[0168]

[0169] wherein,

[0170] R1is C5-C29alkyl or C5-C29alkenyl containing 1 to 5 degrees of unsaturation or is a substituent selected from the group consisting of:

[0171]

[0172] R2is C5-C29alkyl or C5-C29alkenyl containing 1 to 5 degrees of unsaturation or is a substituent selected from the group consisting of:

[0173]

[0174] R A is selected from the group consisting of C5-C29alkyl, C5-C29alkenyl containing 1 to 5 degrees of unsaturation,

[0175] R B is H or is 1 to 2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0176] X1or X2is -O- or -NH-; and X1and X2are not simultaneously -NH-;

[0177] X3is -O- or -NH-;

[0178] R3is selected from the group consisting of H, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C1-C6haloalkoxy;

[0179] R4is H or

[0180] with the additional proviso that:

[0181] R1or R2is not

[0182] and the compound is not

[0183] In another preferred embodiment, the compound of Formula I is a compound of Formula II-A,

[0184]

[0185] wherein each group is as defined above.

[0186] with the additional proviso that:

[0187] when R1is C5-C29alkenyl containing 1-5 degrees of unsaturation, R2is not C5-C29alkyl or C5-C29alkenyl containing 1-5 degrees of unsaturation.

[0188] In another preferred embodiment, the compound is a compound of Formula II-B1 or II-B2,

[0189]

[0190] wherein each group is as defined above.

[0191] In another preferred embodiment, the compound is a compound of Formula III,

[0192]

[0193] wherein each group is as defined above.

[0194] In another preferred embodiment, the compound is a compound of Formula III, A B each of R1, R2, R3, R4, R

[0195] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed between a positively charged group on a compound of Formula (I) and an anion, or a salt formed between a negatively charged group on a compound of Formula (I) and a cation. Suitable anions include, but are not limited to, chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methylsulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronate, lactate, glutarate, or maleate, and the like. Suitable cations include, but are not limited to, sodium ion, potassium ion, magnesium ion, calcium ion, ammonium ion, and the like.

[0196] ​In another preferred embodiment, the pharmaceutically acceptable salt of the present application refers to the salt of the compound of formula (I) with an acid such as, but not limited to, hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, sulfamic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, and isethionic acid, or the salt of the compound of formula (I) with an inorganic base such as, but not limited to, sodium salt, potassium salt, calcium salt, aluminum salt, or ammonium salt, or the salt of the compound of formula (I) with an organic base such as, but not limited to, methylamine salt, ethylamine salt, ethanolamine salt, TRIS ammonium salt, and the like.

[0197] The present application specifically describes the preparation of the compounds of formula (I) by the following methods, which are not intended to limit the scope of the present application. The compounds of the present application can also be readily prepared by combining various synthetic methods described in the present specification or known in the art, which can be easily performed by one skilled in the art.

[0198] Typically, the preparation of the compounds of the present application is carried out by the following procedures, in which the starting materials and reagents used, if not specifically mentioned, are commercially available.

[0199] The present application provides a method for preparing a compound of formula II-A, and the specific steps are as follows:

[0200]

[0201] (S1) In the presence of a base, the compound of formula 1a is subjected to an esterification reaction with a fatty acid having different or the same substituents of R1 and R2, thereby obtaining intermediates 3a-1 to 3a-30;

[0202] (S2) In an inert solvent, the intermediates 3a-1 to 3a-30 are subjected to a deprotection reaction by TFA treatment, thereby obtaining intermediates 4a-1 to 4a-30;

[0203] (S3) In the presence of an inert solvent and nitrogen, the intermediates 4a-1 to 4a-30 are coupled with serine tert-butyl ester and tert-butyl tetraisopropyl phosphorodiamidate to form phosphite triester intermediates 5a-1 to 5a-30;

[0204] (S4) After the intermediates 5a-1 ~ 5a-30 are oxidized in an inert solvent to obtain intermediates 6a-1 ~ 6a-30, the intermediates 6a-1 ~ 6a-30 are deprotected to obtain compounds A1 ~ A30.

[0205] wherein each group is as defined above.

[0206] In another preferred embodiment, the compounds of B1, B3 ~ B7 are prepared by the following method:

[0207]

[0208] (S1) The compound of formula 1b is esterified with a fatty acid having different or the same substituents of R1 and R2 in the presence of a base to obtain intermediates 3b-1 ~ 3b-6.

[0209] (S2) The intermediates 3b-1 ~ 3b-6 are deprotected by reaction with boron chloride in an inert solvent to obtain intermediates 4b-1 ~ 4b-6.

[0210] (S3) The intermediates 4b-1 ~ 4b-6 are subjected to condensation, oxidation and deprotection to obtain compounds B1, B3 ~ B7.

[0211] wherein each group is as defined above.

[0212] Pharmaceutical compositions and methods of administration

[0213] The pharmaceutical composition of the present application comprises a safe and effective amount of the compound of the present application or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. The "safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without producing serious side effects. Generally, the pharmaceutical composition contains 1-2000 mg of the compound of the present application per dose, more preferably, 5-1000 mg of the compound of the present application per dose. Preferably, the "dose" is a capsule or a tablet.

[0214] The "pharmacologically acceptable carrier" means one or more compatible solid or liquid filler or gel materials that are suitable for human use and must be of sufficient purity and sufficiently low toxicity. "Compatible" here means that the components of the composition are capable of being combined with the compound of the present application and with each other in the composition, without chemically interacting in a deleterious manner with the compound. Examples of suitable pharmacologically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyhydric alcohols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as lecithin, acacia, etc.), and the like. coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

[0215] The pharmaceutical composition is an injection, a capsule, a tablet, a pill, a powder, or a granule.

[0216] The mode of administration of the compounds or pharmaceutical compositions of the present application is not particularly limited, and representative modes of administration include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.

[0217] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, such as glycerol; (d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; I retardants, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) absorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage form can also comprise buffering agents.

[0218] Solid dosage forms, such as tablets, dragees, capsules, pills, and granules, can be prepared using a coating and a shell material, such as enteric coatings and other materials well known in the art. They can contain opacifying agents, and can release the active compound or compounds in a certain part of the digestive tract. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compound can also be in micro-encapsulated form, if desired, with one or more of the excipients described above.

[0219] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or elixirs. In addition to the active compound, the liquid dosage forms can include inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and the like, either with or without the addition of

[0220] In addition to these inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0221] In addition to the active compounds, the suspensions can contain suspending agents, e.g., ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide gel, and agar-agar, or mixtures of these substances.

[0222] The compositions for parenteral injection can contain physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyol, and suitable mixtures thereof.

[0223] Dosage forms for topical administration of a compound of this application include ointments, powders, sprays, and inhalers. The active component is admixed with a carrier, which can be a sterile, physiologically acceptable diluent, such as water, ethanol, or a suitable combination of both.

[0224] The compounds of the present application can be administered alone or in combination with other pharmaceutically acceptable compounds (e.g., AD treating agents).

[0225] The therapeutic methods of the present application can be administered alone or in combination with other therapeutic procedures or agents.

[0226] When used as pharmaceuticals, safe and effective amounts of the compounds of the present application are administered to a mammal (e.g., a human) in need of treatment in a pharmaceutically effective amount, which is a dose that is pharmaceutically effective to administer, and is typically in the range of 1 to 2000 mg, preferably 5 to 1000 mg, per day for a 60 kg body weight. Of course, the specific dose will also be determined by the route of administration, the health condition of the patient, and like factors, all within the skill of the skilled practitioner.

[0227] The main advantages of the present application compared to the prior art are:

[0228] (1) The compounds of the present application exhibit good safety to normal epithelial cells Beas-2B and hamster ovary cells CHO.

[0229] (2) The compounds of the present application exhibit good neurotrophic activity and injury protection activity to SH-SY5Y cells.

[0230] (3) The phosphatidylserine derivative A24 of the present application improves the motor ability of Parkinson's disease mice and the cognitive ability of Alzheimer's disease mice.

[0231] The application will be further described in conjunction with specific examples. It should be understood, however, that these examples are intended to illustrate the application and are not intended to limit the scope of the application. The experimental procedures in the following examples, unless otherwise indicated, were generally performed according to routine conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Percentages and parts are percentages by weight and parts by weight, unless otherwise indicated.

[0232] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Also, any methods and materials similar or equivalent to those described herein can be used in the practice of the present application. The preferred methods and materials are described herein.

[0233] In the present application, the structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR determination is performed by using a Bruker AVANCE-400 NMR spectrometer. The LCMS determination is performed by using a Waters 2695 liquid chromatography-mass spectrometry (MS model: Micromass ZQ).

[0234] The HPLC determination is performed by using an Agilent 1100 high pressure liquid chromatograph (ZORBAX SB-C18 250 x 4.6 mm column, Eclipse Plus-C18 250 x 4.6 mm column).

[0235] Compound Examples

[0236] Synthesis of compound A1 of Example 1

[0237] (R)-3-(trityloxy)propane-1,2-diol 1a

[0238]

[0239] (R)-2-((Trityloxy)methyl)oxirane (25.00 g, 79.00 mmol, 1.00 eq.) was dissolved in DMSO (250 mL). A 1.00 M aqueous solution of KOH (129 mL, 1.64 eq.) was added and stirred at 90°C for 12 h. The reaction was monitored by TLC. After completion, the reaction solution was diluted with water (500 mL) and extracted with EA (500 mL x 3). The combined organic layers were washed with brine (500 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 4 / 1 to 1 / 1) to obtain (R)-3-(trityloxy)propane-1,2-diol (15.5 g, 46.4 mmol, 59% yield) as a white solid. 1 H NMR (400MHz, CDCl3) δ=7.47(br d,J=7.6Hz,6H),7.39-7.28(m,9H),3.94-3.86(m,1H),3.79-3.57(m,2H),3.35-3.23(m,2H),2.58(br s,1H),2.09(br s,1H).HRMS(ESI):calcd for C 22 H 22 O3[M+Na] + 357.1461, found 357.1458.

[0240] (S)-3-(trityloxy)propane-1,2-diyl bis(4-heptylbenzoate)3a-1

[0241]

[0242] 4-Heptylbenzoic acid (2.90 g, 13.17 mmol, 2.20 eq.) and 1a (2.00 g, 5.98 mmol, 1.00 eq.) were dissolved in 50 mL of dichloromethane. The reaction system was placed in an ice bath, and DCC (3.71 g, 17.96 mmol, 3.00 eq.) was added and stirred for ten minutes. Subsequently, DMAP (0.37 g, 2.99 mmol, 0.50 eq.) was added, and the mixture was stirred at 0°C for 2 hours. Completion of the reaction was checked by TLC. After completion, the reaction mixture was filtered through celite and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 20 / 1 to 5 / 1) to obtain compound 3a-1 as a colorless, transparent oil in a 72% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.00 (d, J = 7.9 Hz, 2H), 7.82 (d, J = 8.0 Hz, 2H), 7.48 - 7.42 (m, 6H), 7.21 (dd, J = 16.5, 8.2 Hz, 13H), 5.62 (p, J = 4.9 Hz, 1H), 4.67 (h, J = 5.1, 4.4 Hz, 2H), 3.45 (ddd, J = 31.4, 10.0, 4.7 Hz, 2H), 2.65 (q, J = 8.2 Hz, 4H), 1.62 (d, J = 7.5 Hz, 5H), 1.31 (p, J = 6.0, 4.7 Hz, 16H), 0.90 - 0.86 (m, 6H). HRMS (ESI): calcd for C 50 H 58 O5[M+Na] + 519.3081, found 519.3074.

[0243] (S)-3-hydroxypropane-1,2-diyl bis(4-heptylbenzoate)4a-1

[0244]

[0245] Intermediate 3a-1 was dissolved in dichloromethane (10 mL), and a solution of TFA (5 mL) was added slowly. The reaction was stirred at room temperature for 2 h. After the reaction was detected by TLC plate, an appropriate amount of NaHCO3(aq.) solution was added to adjust the pH of the reaction system to 7-9. Then the residual solution was extracted with EA (30 mL x 3), the organic layers were combined and washed with brine (30 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 10 / 1-3 / 1) to obtain compound 4a-1 as a white solid in a yield of 51%. 1 H NMR (400 MHz, Chloroform-d) δ 7.96 (dd, J = 12.4, 7.9 Hz, 4H), 7.26 - 7.22 (m, 4H), 5.44 (d, J = 5.1 Hz, 1H), 4.65 (d, J = 5.0 Hz, 2H), 3.94 (d, J = 5.0 Hz, 2H), 2.68 - 2.63 (m, 4H), 1.67 - 1.24 (m, 21H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 31 H 44 O5[M+Na] + 519.3081, found 519.3074. 519.3081, found 519.3074.

[0246] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)propane-1,2-diyl bis(4-heptylbenzoate)5a-1

[0247]

[0248] Dissolve 1-tert-butoxy-N,N,N',N'-tetraisopropylphosphorodiamidite (813 mg, 2.67 mmol, 1.00 eq.) in dichloromethane (10.0 mL) and toluene (1.00 mL). To this solution, add tert-butyl (tert-butoxycarbonyl)-D-serine (488 mg, 1.87 mmol, 0.70 eq.). To eliminate trace amounts of water, evaporate the resulting solution under vacuum. Dissolve the residue in anhydrous dichloromethane (10.0 mL) under nitrogen protection, add 1H-tetrazole (168 mg, 2.40 mmol, 0.90 eq.) at room temperature, and after a few minutes, a white solid precipitates. Stir the whole under nitrogen at room temperature for 3 h. Add 4a-1 (993 mg, 2.00 mmol, 0.75 eq.) and 1H-tetrazole (374 mg, 5.34 mmol, 2.00 eq.) dissolved in dichloromethane (3.00 mL) under nitrogen. Then, stir the mixture at room temperature for 11 hours. Check the reaction completion with TLC plate, after the reaction is completed, quench the reaction mixture with sodium bicarbonate solution (30.0 mL), then extract with dichloromethane (30.0 mL x 3). Wash the combined organic layers with brine (30.0 mL x 2), dry over sodium sulfate, concentrate under reduced pressure to get a residue. Purify the residue by silica gel column (PE / EA = 20 / 1 ~ 10 / 1, 0.1% TEA) to get compound 5a-1 as colorless transparent oil, yield 27%. 1 H NMR (400 MHz, Chloroform-d) δ 7.86 (ddd, J = 15.9, 11.8, 8.1 Hz, 4H), 7.36 - 7.27 (m, 4H), 6.90 (dd, J = 13.9, 7.2 Hz, 1H), 5.49 (t, J = 5.3 Hz, 1H), 4.83 - 3.76 (m, 6H), 2.68 - 2.58 (m, 4H), 1.56 (q, J = 7.3 Hz, 4H), 1.40 - 1.30 (m, 20H), 1.32 - 1.19 (m, 24H), 0.84 (tt, J = 6.5, 3.1 Hz, 6H). HRMS (ESI): calcd for C 47 H74 NO 11 P[M+Na] + 882.4892, found 882.4896.

[0249] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)propane-1,2-diylbis(4-heptylbenzoate)6a-1

[0250]

[0251] To a solution of 5a-1 (439 mg, 0.51 mmol, 1.00 eq.) in dichloromethane (10.0 mL) was added tert-butyl hydroperoxide (132 mg, 1.02 mmol, 70% purity, 2.00 eq.) and stirred at room temperature for 1 h. The reaction was monitored by TLC plate. After the reaction was completed, water (10.0 mL) was added to quench the reaction and extracted with dichloromethane (10.0 mL x 3). The combined organic layers were washed with brine (20.0 mL x 3), dried over sodium sulfate, filtered and concentrated to give a residue. The residue was purified by silica gel column chromatography (PE / EA = 5 / 1 ~ 2 / 1, 0.1% TEA) to give compound 6a-1 as colorless transparent oil in 55% yield. 1 H NMR (400 MHz, Methanol-d4) δ 7.98 - 7.87 (m, 4H), 7.25 (t, J = 7.6 Hz, 4H), 5.64 (p, J = 5.1 Hz, 1H), 4.72 - 4.54 (m, 2H), 4.46 - 4.19 (m, 5H), 2.64 (td, J = 7.7, 4.5 Hz, 4H), 1.60 (dt, J = 10.3, 4.6 Hz, 4H), 1.44 (dd, J = 9.6, 6.4 Hz, 27H), 1.38 - 1.19 (m, 17H), 0.87 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 47 H 74 NO 12 P[M+Na] + 898.4841, found 898.4834.

[0252] O-(((R)-2,3-bis((4-heptylbenzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A1

[0253]

[0254] A solution of 6a-1 (244 mg, 0.28 mmol, 1.00 eq.) in trifluoroacetic acid (3.50 mL) was stirred at 0 °C. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was dissolved in dichloromethane (3.00 mL x 3) and concentrated under reduced pressure to remove excess trifluoroacetic acid. The residue was dissolved in acetonitrile (10.00 mL), ultrasonically shaken and slurry, and the solution was filtered off and pumped dry to obtain compound A1 as a white solid in a yield of 50%. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.88 (dd, J = 8.0, 15.2 Hz, 4H), 7.76 (br s, 3H), 7.24 (dd, J = 5.2, 7.6 Hz, 4H), 5.66 (br s, 1H), 4.77-4.67 (m, 1H), 4.58 (br dd, J = 6.4, 11.6 Hz, 3H), 4.49-4.41 (m, 1H), 4.35 (br d, J = 5.6 Hz, 2H), 2.64 (br t, J = 7.6 Hz, 4H), 1.61 (br d, J = 6.4 Hz, 4H), 1.34-1.24 (m, 16H), 0.88 (br t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDCl3 and 20 μL TFA) δ = 168.85, 168.70, 168.25, 150.73, 150.50, 130.00, 129.90, 128.82, 128.80, 125.50, 125.38, 70.98, 70.91, 65.21, 63.64, 63.24, 54.26, 54.19, 36.04, 31.71, 30.96, 29.18, 29.05, 22.59, 13.95. HRMS (ESI): calcd for C 34 H 50 NO 10 P[M+H] + 664.3245, found 664.3237.

[0255] Synthesis of compound A2 of example 2

[0256] (S)-3-(trityloxy)propane-1,2-diyl bis(4-(hexyloxy)benzoate)3a-2

[0257]

[0258] The procedure described in 3a-1 was followed using 1a and 4-hexyloxybenzoic acid to give the target compound as a colorless transparent oil in 38% yield. 1 H NMR (400 MHz, Methanol-d4) δ 8.01 - 7.97 (m, 2H), 7.81 - 7.78 (m, 2H), 7.44 - 7.40 (m, 6H), 7.26 - 7.19 (m, 9H), 7.00 - 6.95 (m, 2H), 6.92 - 6.88 (m, 2H), 5.54 (p, J = 5.1 Hz, 1H), 4.61 (dd, J = 5.3, 1.9 Hz, 2H), 4.03 (q, J = 6.6 Hz, 4H), 3.43 (qd, J = 10.1, 4.8 Hz, 2H), 1.78 (dtd, J = 14.4, 6.4, 1.9 Hz, 4H), 1.48 (dd, J = 11.4, 8.3 Hz, 4H), 1.39 - 1.30 (m, 8H), 0.92 (tt, J = 5.4, 2.4 Hz, 6H). HRMS (ESI): calcd for C 48 H 54 O7[M+Na] + 765.3762, found 765.3751.

[0259] (S)-3-hydroxypropane-1,2-diyl bis(4-(hexyloxy)benzoate)4a-2

[0260]

[0261] The procedure described in 4a-1 was followed using 3a-2 to give the target compound as a white solid in 64% yield. 1 H NMR (400 MHz, Methanol-d4) δ 8.00 - 7.85 (m, 4H), 6.98 - 6.87 (m, 4H), 5.41 (dtd, J = 6.7, 5.1, 3.7 Hz, 1H), 4.69 - 4.48 (m, 2H), 4.00 (td, J = 6.5, 4.2 Hz, 4H), 3.88 (d, J = 5.2 Hz, 2H), 1.82 - 1.71 (m, 4H), 1.50 - 1.41 (m, 4H), 1.34 (qt, J = 4.4, 2.5 Hz, 8H), 0.97 - 0.85 (m, 6H). HRMS (ESI): calcd for C 29 H 40 O7[M+Na] + 523.2666, found 523.2667.

[0262] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(4-(hexyloxy)benzoate) 5a-2

[0263]

[0264] Following the procedure described in 5a-1, 4a-2 was treated to give the target compound as a colorless transparent oil in 38% yield. 1 HNMR (400 MHz, Methanol-d4) δ 8.01 - 7.83 (m, 4H), 6.99 - 6.82 (m, 4H), 6.51 (d, J = 8.4 Hz, 1H), 5.54 - 5.42 (m, 1H), 4.67 - 4.41 (m, 2H), 4.27 - 3.84 (m, 8H), 1.77 (dtd, J = 14.5, 6.4, 2.0 Hz, 4H), 1.52 - 1.40 (m, 27H), 1.40 - 1.32 (m, 12H), 0.96 - 0.88 (m, 6H). HRMS (ESI): calcd for C 45 H 70 O 13 P[M+Na] + 886.4477, found 886.447.

[0265] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(4-(hexyloxy)benzoate) 5a-2

[0266]

[0267] Following the procedure described in 6a-1, 5a-2 was treated to give the target compound as a colorless transparent oil in 65% yield. 1HNMR (400 MHz, Chloroform-d) δ 8.07 - 7.89 (m, 4H), 6.87 (dddd, J = 13.8, 9.5, 4.9, 2.8 Hz, 4H), 5.56 (pd, J = 10.3, 6.0 Hz, 2H), 4.68 - 4.08 (m, 7H), 4.04 - 3.92 (m, 4H), 1.83 - 1.73 (m, 4H), 1.55 - 1.21 (m, 39H), 0.95 - 0.86 (m, 6H). HRMS (ESI): calcd for C 45 H 70 O 14 P[M+Na] + 902.4426, found 902.4428.

[0268] O-(((R)-2,3-bis((4-(hexyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A2

[0269]

[0270] Following the procedure described for Al, 6a-2 was treated to give the target compound as a light yellow solid in 61% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.75 (br s, 3H), 7.57 (br dd, J = 7.6, 16.8 Hz, 2H), 7.49 (br d, J = 16.4 Hz, 2H), 7.34 (dt, J = 5.2, 8.0 Hz, 2H), 7.16 (br d, J = 2.8 Hz, 2H), 5.67 (br s, 1H), 4.78 - 4.69 (m, 1H), 4.65 - 4.51 (m, 3H), 4.49 - 4.27 (m, 3H), 4.05 - 3.88 (m, 4H), 1.76 (br s, 4H), 1.43 (br s, 4H), 1.27 (s, 40H), 0.89 (br t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 168.89, 168.13, 167.70, 167.20, 164.13, 163.98, 132.17, 132.05, 120.39, 120.22, 120.05, 114.47, 70.83, 70.43, 68.46, 66.06, 65.17, 63.09, 62.73, 31.48, 28.95, 25.57, 25.53, 22.55, 22.52, 13.92. HRMS (ESI): calcd for C 32 H 46 NO 12 P[M+H] + 668.2830, found 668.2833.

[0271] Synthesis of compound A3 of Example 3

[0272] (S)-3-(trityloxy)propane-1,2-diyl bis(3-(hexyloxy)benzoate)3a-3

[0273]

[0274] Following the procedure described in 3a-1, 1a and 3-hexyloxybenzoic acid were treated to give the target compound as colorless transparent oil in 50% yield. 1 H NMR (400 MHz, Methanol-d4) δ 7.63 (dt, J = 7.7, 1.3 Hz, 1H), 7.55 (dd, J = 2.7, 1.5 Hz, 1H), 7.48 - 7.42 (m, 7H), 7.41 - 7.35 (m, 2H), 7.31 (t, J = 8.0 Hz, 1H), 7.29 - 7.08 (m, 11H), 5.65 - 5.58 (m, 1H), 4.71 (dd, J = 11.7, 4.2 Hz, 1H), 4.62 (dd, J = 11.8, 6.3 Hz, 1H), 3.91 (dt, J = 21.6, 6.4 Hz, 4H), 3.50 - 3.43 (m, 2H), 1.74 (dtd, J = 14.6, 6.4, 2.2 Hz, 4H), 1.49 - 1.29 (m, 12H), 0.96 - 0.84 (m, 6H). HRMS (ESI): calcd for C 48 H 54 O7[M+Na] + 765.3762, found 765.3753.

[0275] (S)-3-hydroxypropane-1,2-diyl bis(3-(hexyloxy)benzoate)4a-3

[0276]

[0277] Following the procedure described in 4a-1, 3a-3 was treated to give the target compound as a white solid in 68% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.62 (ddt, J = 13.8, 7.6, 1.3 Hz, 2H), 7.54 (ddd, J = 12.5, 2.6, 1.5 Hz, 2H), 7.32 (td, J = 7.9, 5.5 Hz, 2H), 7.10 (dddd, J = 8.0, 3.8, 2.6, 1.1 Hz, 2H), 5.46 (p, J = 5.0 Hz, 1H), 4.71 - 4.60 (m, 2H), 3.96 (dt, J = 8.5, 6.5 Hz, 6H), 2.29 (s, 1H), 1.77 (dtt, J = 10.3, 6.5, 3.8 Hz, 4H), 1.44 (dddd, J = 9.5, 7.0, 4.8, 2.5 Hz, 4H), 1.37 - 1.30 (m, 8H), 0.95 - 0.85 (m, 6H). HRMS (ESI): calcd for C 29 H 40 O7[M+Na] + 523.2666, found 523.2658.

[0278] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)propane-1,2-diyl bis(3-(hexyloxy)benzoate)5a-3

[0279]

[0280] Following the procedure described in 5a-1, 4a-3 was treated to give the target compound as a colorless transparent oil in 50% yield. 1HNMR (400 MHz, DMSO-d6) δ 7.52 (dddd, J = 12.7, 7.7, 2.6, 1.2 Hz, 2H), 7.43 - 7.36 (m, 4H), 7.20 (dddd, J = 7.9, 4.8, 2.9, 1.8 Hz, 2H), 6.90 (dd, J = 8.1, 4.9 Hz, 1H), 5.51 (q, J = 5.1, 4.5 Hz, 1H), 4.67 (dt, J = 11.9, 3.2 Hz, 1H), 4.50 (dd, J = 11.9, 6.8 Hz, 1H), 4.20 - 3.83 (m, 9H), 1.72 - 1.64 (m, 4H), 1.41 - 1.26 (m, 39H), 0.87 (td, J = 5.8, 3.5 Hz, 6H). HRMS (ESI): calcd for C 45 H 70 NO 13 P[M+Na] + 886.4477, found 886.4498.

[0281] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(3-(hexyloxy)benzoate)6a-3

[0282]

[0283] Following the procedure described in 6a-1, 5a-3 was treated to give the target compound as a colorless transparent oil in 27% yield. 1HNMR (400 MHz, Chloroform-d) δ 7.61 (ddt, J = 15.1, 7.7, 1.4 Hz, 2H), 7.59 - 7.48 (m, 2H), 7.31 (td, J = 7.9, 3.3 Hz, 2H), 7.13 - 7.05 (m, 2H), 5.61 (t, J = 5.1 Hz, 1H), 5.54 (t, J = 10.8 Hz, 1H), 4.69 (ddd, J = 12.0, 6.5, 4.1 Hz, 1H), 4.56 (dt, J = 11.8, 5.8 Hz, 1H), 4.41 - 4.34 (m, 0H), 4.36 (s, 2H), 4.37 - 4.31 (m, 1H), 4.34 - 4.18 (m, 1H), 3.95 (q, J = 6.4 Hz, 4H), 1.77 (dtd, J = 14.9, 6.5, 3.2 Hz, 4H), 1.50 - 1.41 (m, 29H), 1.39 - 1.32 (m, 4H), 1.35 - 1.23 (m, 5H), 1.27 (s, 1H), 0.91 (td, J = 6.8, 5.8, 3.2 Hz, 6H). HRMS (ESI): calcd for C 45 H 70 NO 14 P[M+Na] + 902.4426, found 902.4424.

[0284] O-(((R)-2,3-bis((3-(hexyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A3

[0285]

[0286] Following the procedure described for Al, 6a-3 was treated to give the target compound as a light yellow solid in 45% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.80 (br s, 3H), 7.63 - 7.43 (m, 4H), 7.40 - 7.28 (m, 2H), 7.14 (br s, 2H), 5.66 (br d, J = 1.2 Hz, 1H), 4.72 (br d, J = 11.6 Hz, 1H), 4.56 (br d, J = 5.2 Hz, 3H), 4.47 - 4.23 (m, 3H), 4.04 - 3.85 (m, 4H), 1.76 (br d, J = 5.2 Hz, 4H), 1.43 (br s, 4H), 1.33 (br d, J = 2.8 Hz, 8H), 0.90 (br s, 6H).13 C NMR (101 MHz, CDCl3 and 20 μL TFA) δ = 168.79, 167.86, 167.38, 159.05, 159.01, 129.68, 129.61, 122.22, 122.15, 121.05, 115.34, 115.33, 115.08, 115.06, 71.01, 70.85, 68.58, 68.53, 65.11, 63.66, 63.21, 54.14, 31.48, 28.97, 25.54, 22.52, 13.89. HRMS (ESI): calcd for C 32 H 46 NO 12 P[M+H] + 668.2830, found 668.2825.

[0287] Synthesis of compound A4 of example 4

[0288] (S)-2-hydroxy-3-(trityloxy)propyl 4-heptylbenzoate 2a-1

[0289]

[0290] Following the procedure described for 3a-1, 4-heptylbenzoic acid (1.45 g, 6.58 mmol, 1.10 eq.) and 1a (2 g, 5.98 mmol, 1.00 eq.) were treated to give the target compound as a colorless transparent oil in 64% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.78 (d, J = 8.0 Hz, 1H), 7.36 (d, J = 7.4 Hz, 6H), 7.17 (dt, J = 24.2, 7.4 Hz, 11H), 4.35 (dd, J = 5.3, 3.3 Hz, 1H), 4.03 (q, J = 7.1, 6.6 Hz, 1H), 3.24 (d, J = 5.2 Hz, 2H), 2.62 - 2.53 (m, 2H), 1.54 (p, J = 7.3 Hz, 2H), 1.29 - 1.12 (m, 8H), 0.84 - 0.74 (m, 3H). HRMS (ESI): calcd for C 36 H 40 O4[M+Na] + 559.2819, found 559.2822.

[0291] (S)-2-(decanoyloxy)-3-(trityloxy)propyl 4-heptylbenzoate 3a-4

[0292]

[0293] Decanoic acid (0.93 g, 4.48 mmol, 1.20 eq.) and 2a-1 (2.00 g, 3.73 mmol, 1.00 eq.) were treated according to the procedure described for 3a-1 to give the target compound as a colorless transparent oil in 96% yield. 1 H NMR (400 MHz, DMSO-d6) δ 7.77 - 7.66 (m, 2H), 7.44 - 7.18 (m, 17H), 5.34 (dp, J = 9.7, 4.9 Hz, 1H), 4.51 - 4.32 (m, 2H), 3.23 (d, J = 5.2 Hz, 2H), 2.63 (t, J = 7.6 Hz, 2H), 2.30 (t, J = 7.2 Hz, 2H), 1.53 (dp, J = 34.3, 7.2 Hz, 4H), 1.32 - 1.07 (m, 22H), 0.83 (dt, J = 9.9, 6.8 Hz, 6H). HRMS (ESI): calcd for C 46 H 58 O5[M+Na] + 713.4176, found 713.4186.

[0294] (S)-2-(decanoyloxy)-3-hydroxypropyl 4-heptylbenzoate 4a-4

[0295]

[0296] 3a-4 was treated according to the procedure described for 4a-1 to give the target compound as a white solid in 72% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.95 (dd, J = 8.2, 2.5 Hz, 2H), 7.24 (dd, J = 8.2, 2.6 Hz, 2H), 4.40 (d, J = 4.0 Hz, 2H), 4.33 - 4.19 (m, 3H), 2.97 - 2.90 (m, 1H), 2.65 (t, J = 7.4 Hz, 2H), 2.35 (td, J = 7.7, 2.7 Hz, 2H), 1.62 (t, J = 7.4 Hz, 4H), 1.28 (dd, J = 14.5, 6.4 Hz, 20H), 0.91 - 0.84 (m, 6H). HRMS (ESI): calcd for C 27H 44 O5[M+Na] + 471.3081, found 471.308.

[0297] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(decanoyloxy)propyl 4-heptylbenzoate5a-4

[0298]

[0299] According to the method described in 5a-1, 4a-4 was treated to obtain the target compound, colorless transparent oil, yield 42%. 1 HNMR (400 MHz, DMSO-d6) δ 7.89 (d, J = 8.0 Hz, 2H), 7.34 (d, J = 8.0 Hz, 2H), 6.87 (dd, J = 13.3, 8.0 Hz, 1H), 4.65 - 4.54 (m, 1H), 4.36 (s, 2H), 4.21 (d, J = 5.4 Hz, 2H), 4.07 - 3.80 (m, 3H), 2.64 (t, J = 7.7 Hz, 2H), 2.30 (td, J = 7.4, 2.7 Hz, 2H), 1.64 - 1.41 (m, 4H), 1.41 - 1.33 (m, 26H), 1.32 - 1.09 (m, 21H), 0.88 - 0.80 (m, 6H). HRMS (ESI): calcd for C 43 H 74 NO 11 P[M+Na] + 834.4892, found 834.4881.

[0300] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(decanoyloxy)propyl 4-heptylbenzoate6a-4

[0301]

[0302] According to the method described in 6a-1, 5a-4 was treated to obtain the target compound, colorless transparent oil, yield 80%. 1HNMR (400 MHz, Chloroform-d) δ 7.95 (dd, J = 8.3, 1.6 Hz, 2H), 7.24 (dd, J = 8.3, 2.8 Hz, 2H), 5.58 (dd, J = 8.2, 4.1 Hz, 1H), 4.82 (dtt, J = 9.1, 6.3, 3.2 Hz, 1H), 4.56 - 4.21 (m, 6H), 2.64 (t, J = 7.7 Hz, 2H), 2.33 (q, J = 8.0 Hz, 2H), 1.61 (d, J = 6.8 Hz, 4H), 1.51 - 1.19 (m, 47H), 0.86 (td, J = 6.9, 1.7 Hz, 6H). HRMS (ESI): calcd for C 43 H 74 NO 12 P[M+Na] + 850.4841, found 850.4836.

[0303] O-(((R)-2-(decanoyloxy)-3-((4-heptylbenzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A4

[0304]

[0305] Following the procedure described for Al, 6a-4 was treated to give the target compound as a light yellow solid in 50% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.88 (br d, J = 8.0 Hz, 2H), 7.75 (br s, 3H), 7.27 (br s, 2H), 4.86 - 4.76 (m, 1H), 4.60 (br dd, J = 4.8, 16.4 Hz, 3H), 4.53 - 4.40 (m, 3H), 4.34 (br dd, J = 4.8, 11.6 Hz, 1H), 2.65 (br t, J = 7.6 Hz, 2H), 2.39 (br t, J = 7.6 Hz, 2H), 1.60 (br d, J = 7.2 Hz, 4H), 1.36 - 1.22 (m, 20H), 0.93 - 0.84 (m, 6H). 13C NMR (101 MHz, CDCl3 and 20 μL TFA) δ = 176.87, 168.87, 168.51, 150.64, 129.89, 128.82, 125.45, 73.22, 73.17, 63.62, 63.33, 54.18, 36.04, 34.11, 31.77, 30.96, 29.29, 29.19, 29.05, 28.92, 24.66, 22.59. HRMS (ESI): calcd for C 30 H 50 NO 10 P[M+H] + 616.3245, found 616.3245.

[0306] Synthesis of compound A5 in Example 5

[0307] (S)-2-hydroxy-3-(trityloxy)propyl 4-(hexyloxy)benzoate 2a-2

[0308]

[0309] Following the procedure described in 2a-1, 4-hexyloxybenzoic acid and 1a were treated to give the target compound as a colorless transparent oil in 47% yield. 1 H NMR (400 MHz, Methanol-d4) δ 7.89 - 7.83 (m, 2H), 7.44 (dd, J = 7.8, 1.7 Hz, 6H), 7.25 (dd, J = 8.3, 6.5 Hz, 6H), 7.22 - 7.15 (m, 4H), 6.95 - 6.88 (m, 2H), 4.39 (d, J = 5.1 Hz, 2H), 4.05 (dt, J = 17.6, 5.9 Hz, 3H), 3.25 (dd, J = 5.6, 2.4 Hz, 2H), 1.79 (dt, J = 14.7, 6.6 Hz, 2H), 1.49 (qt, J = 10.0, 6.5 Hz, 2H), 1.36 (h, J = 4.9, 4.2 Hz, 4H), 0.96 - 0.88 (m, 3H). HRMS (ESI): calcd for C 35 H 38 NO5[M+Na] + 561.2611, found 561.2614.

[0310] (S)-2-(decanoyloxy)-3-(trityloxy)propyl 4-(hexyloxy)benzoate 3a-5

[0311]

[0312] The target compound was obtained by treating decanoic acid and 2a-2 according to the method described in 3a-4, colorless transparent oil, yield 80%. 1 H NMR (400 MHz, DMSO-d6) δ 7.76 - 7.70 (m, 2H), 7.39 - 7.23 (m, 15H), 7.02 - 6.94 (m, 2H), 5.32 (dt, J = 9.3, 4.7 Hz, 1H), 4.49 - 4.36 (m, 2H), 4.03 (t, J = 6.5 Hz, 2H), 3.22 (d, J = 5.1 Hz, 2H), 2.30 (t, J = 7.1 Hz, 2H), 1.77 - 1.65 (m, 2H), 1.53 - 1.35 (m, 4H), 1.34 - 1.05 (m, 16H), 0.91 - 0.77 (m, 6H). HRMS (ESI): calcd for C 45 H 56 O6[M+Na] + 715.3925, found 715.392.

[0313] (S)-2-(decanoyloxy)-3-hydroxypropyl 4-(hexyloxy)benzoate 4a-5

[0314]

[0315] The target compound was obtained by treating 3a-5 according to the method described in 4a-1, white solid, yield 56%. 1 H NMR (400 MHz, DMSO-d6) δ 7.94 - 7.89 (m, 2H), 7.05 - 6.97 (m, 2H), 5.38 (s, 1H), 4.20 (dd, J = 5.3, 1.2 Hz, 2H), 4.08 (d, J = 5.4 Hz, 2H), 4.02 (t, J = 6.5 Hz, 3H), 2.27 (t, J = 7.4 Hz, 2H), 1.75 - 1.65 (m, 2H), 1.55 - 1.34 (m, 4H), 1.33 - 1.12 (m, 16H), 0.90 - 0.79 (m, 6H). HRMS (ESI): calcd for C 26 H 42 O6[M+H] + 451.3054, found 451.3066.

[0316] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(decanoyloxy)propyl 4-(hexyloxy)benzoate 5a-5

[0317]

[0318] Following the procedure described in 5a-1, 4a-5 was treated to give the target compound as a colorless transparent oil in 33% yield. 1 HNMR (400 MHz, Methanol-d4) δ 8.02 - 7.94 (m, 2H), 7.00 - 6.90 (m, 2H), 4.70 - 4.54 (m, 1H), 4.47 - 3.95 (m, 8H), 2.33 (td, J = 7.4, 2.2 Hz, 2H), 1.84 - 1.72 (m, 2H), 1.60 (h, J = 7.1, 6.6 Hz, 2H), 1.50 - 1.20 (m, 45H), 0.94 - 0.85 (m, 6H). HRMS (ESI): calcd for C 42 H 72 NO 12 P[M+Na] + 836.4684, found 836.4685.

[0319] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(decanoyloxy)propyl 4-(hexyloxy)benzoate 5a-5

[0320]

[0321] Following the procedure described in 6a-1, 5a-5 was treated to give the target compound as a colorless transparent oil in 62% yield. 1HNMR (400 MHz, Chloroform-d) δ 8.05 - 7.89 (m, 2H), 6.95 - 6.79 (m, 2H), 5.60 (d, J = 7.8 Hz, 1H), 4.84 (dp, J = 9.5, 4.9 Hz, 1H), 4.75 - 4.07 (m, 9H), 3.98 (dt, J = 25.0, 6.6 Hz, 2H), 2.43 - 2.21 (m, 2H), 1.79 (dq, J = 14.1, 5.8, 5.0 Hz, 2H), 1.68 - 1.12 (m, 45H), 0.95 - 0.83 (m, 6H). HRMS (ESI): calcd for C 42 H 72 NO 13 P[M+Na] + 852.4633, found 852.4637.

[0322] O-(((R)-2-(decanoyloxy)-3-((4-(hexyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A5

[0323]

[0324] According to the method described in A1, 6a-5 was treated to give the target compound, pale yellow solid, yield 55%. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.91 (br d, J = 8.8 Hz, 2H), 7.82 (br s, 2H), 6.90 (br d, J = 8.8 Hz, 2H), 4.73 (br s, 1H), 4.64 - 4.37 (m, 6H), 4.31 (br d, J = 7.2 Hz, 1H), 4.00 (br t, J = 6.4 Hz, 2H), 2.37 (br t, J = 7.6 Hz, 2H), 1.87 - 1.74 (m, 2H), 1.58 (br s, 2H), 1.45 (br d, J = 6.4 Hz, 2H), 1.39 - 1.32 (m, 4H), 1.24 (br s, 12H), 0.89 (td, J = 6.8, 17.6 Hz, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.50, 169.12, 168.14, 164.20, 132.08, 119.95, 114.50, 72.91, 68.49, 63.65, 63.18, 54.16, 34.06, 31.77, 31.48, 29.30, 29.16, 29.06, 28.93, 25.53, 24.65, 22.59, 22.52, 13.96, 13.91. HRMS (ESI): calcd for C 29 H 48 NO 11 P[M+H] + 618.3038, found 618.3042.

[0325] Synthesis of compound A6 of Example 6

[0326] (S)-2-hydroxy-3-(trityloxy)propyl 3-(hexyloxy)benzoate 2a-3

[0327]

[0328] Following the procedure described in 2a-1, 3-hexyloxybenzoic acid and la were treated to give the target compound as a colorless transparent oil in 68% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.53 - 7.39 (m, 7H), 7.33 - 7.17 (m, 11H), 7.07 (dd, J = 8.2, 2.9 Hz, 1H), 4.42 (dd, J = 5.4, 2.5 Hz, 2H), 4.11 (s, 1H), 3.95 (t, J = 6.5 Hz, 2H), 3.30 (d, J = 5.1 Hz, 2H), 2.45 (s, 1H), 1.81 - 1.73 (m, 2H), 1.45 (t, J = 7.6 Hz, 2H), 1.32 (dt, J = 7.6, 3.8 Hz, 4H), 0.93 - 0.85 (m, 3H). HRMS (ESI): calcd for C 35 H 38 NO5[M+Na] + 561.2611, found 561.2618.

[0329] (S)-2-(decanoyloxy)-3-(trityloxy)propyl 3-(hexyloxy)benzoate 3a-6

[0330] (S)-2-(decanoyloxy)-3-(trityloxy)propyl 3-(hexyloxy)benzoate 3a-6

[0331] The target compound was obtained by treating decanoic acid and 2a-3 according to the method described in 3a-4, colorless transparent oil, yield 66%. 1 H NMR (400 MHz, Chloroform-d) δ 7.49 - 7.39 (m, 8H), 7.32 - 7.17 (m, 10H), 7.07 (ddd, J = 8.3, 2.6, 1.1 Hz, 1H), 5.45 - 5.37 (m, 1H), 4.58 - 4.45 (m, 2H), 3.95 (t, J = 6.5 Hz, 2H), 3.32 (qd, J = 9.9, 5.1 Hz, 2H), 2.32 (dd, J = 8.1, 6.9 Hz, 2H), 1.83 - 1.73 (m, 2H), 1.66 - 1.55 (m, 2H), 1.47 (q, J = 7.5 Hz, 2H), 1.40 - 1.15 (m, 18H), 0.94 - 0.82 (m, 6H). HRMS (ESI): calcd for C 45 H 56 O6[M+Na] + 715.3925, found 715.3936.

[0332] (S)-2-(decanoyloxy)-3-hydroxypropyl 3-(hexyloxy)benzoate 4a-6

[0333]

[0334] The target compound was obtained by treating 3a-6 according to the method described in 4a-1, white solid, yield 62%. 1 H NMR (400 MHz, Chloroform-d) δ 7.61 (dt, J = 7.7, 1.3 Hz, 1H), 7.55 (dd, J = 2.7, 1.5 Hz, 1H), 7.34 (t, J = 8.0 Hz, 1H), 7.11 (ddd, J = 8.3, 2.7, 1.0 Hz, 1H), 4.42 - 4.40 (m, 1H), 4.33 - 4.18 (m, 3H), 4.00 (t, J = 6.5 Hz, 2H), 2.35 (t, J = 7.6 Hz, 2H), 1.85 - 1.75 (m, 2H), 1.52 - 1.43 (m, 2H), 1.39 - 1.21 (m, 20H), 0.89 (dt, J = 13.8, 7.0 Hz, 6H). HRMS (ESI): calcd for C 26 H 42 O6[M+Na] +473.2874, found 473.2869.

[0335] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(decanoyloxy)propyl 4-(hexyloxy)benzoate 5a-6

[0336]

[0337] Following the procedure described in 5a-1, 4a-6 was treated to give the target compound as a colorless transparent oil in 63% yield. 1 HNMR (400 MHz, DMSO-d6) δ 7.55 (dt, J = 7.6, 1.3 Hz, 1H), 7.49 - 7.39 (m, 2H), 7.26 - 7.21 (m, 1H), 6.84 (dd, J = 21.1, 8.1 Hz, 1H), 4.60 (ddq, J = 15.0, 9.9, 5.0 Hz, 1H), 4.44 - 4.31 (m, 2H), 4.22 (h, J = 6.1, 5.4 Hz, 2H), 4.10 - 3.91 (m, 4H), 2.31 (td, J = 7.3, 2.4 Hz, 2H), 1.78 - 1.65 (m, 2H), 1.57 - 1.11 (m, 49H), 0.92 - 0.80 (m, 6H). HRMS (ESI): calcd for C 42 H 72 NO 12 P[M+Na] + 836.4684, found 836.4688.

[0338] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(decanoyloxy)propyl 3-(hexyloxy)benzoate 6a-6

[0339]

[0340] Following the procedure described in 6a-1, 5a-6 was treated to give the target compound as a colorless transparent oil in 89% yield. 1HNMR (400 MHz, DMSO-d6) δ 7.58 (dq, J = 7.6, 1.2 Hz, 1H), 7.52 (q, J = 2.1 Hz, 1H), 7.44 (td, J = 8.0, 1.1 Hz, 1H), 7.24 (dq, J = 8.3, 2.0, 1.5 Hz, 1H), 7.12 (dd, J = 19.1, 7.6 Hz, 1H), 4.81 (s, 1H), 4.51 - 4.06 (m, 7H), 4.02 (t, J = 6.5 Hz, 2H), 2.33 (td, J = 7.4, 4.8 Hz, 2H), 1.73 (p, J = 6.7 Hz, 2H), 1.61 - 1.15 (m, 47H), 0.87 (dt, J = 13.4, 7.0 Hz, 6H). HRMS (ESI): calcd for C 42 H 72 NO 13 P[M+Na] + 852.4633, found 852.463.

[0341] O-(((R)-2-(decanoyloxy)-3-((3-(hexyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A6

[0342]

[0343] The procedure described in Al was followed to treat 6a-6 to give the target compound as a white solid in 54% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.76 (br s, 3H), 7.57 (br d, J = 7.6 Hz, 1H), 7.51 (br s, 1H), 7.35 (br t, J = 8.0 Hz, 1H), 7.19 - 7.12 (m, 1H), 4.79 (br d, J = 4.0 Hz, 1H), 4.66 - 4.38 (m, 6H), 4.37 - 4.28 (m, 1H), 4.06 - 3.93 (m, 2H), 2.44 - 2.30 (m, 2H), 1.84 - 1.72 (m, 2H), 1.58 (br d, J = 6.8 Hz, 2H), 1.50 - 1.40 (m, 2H), 1.38 - 1.31 (m, 4H), 1.24 (br s, 12H), 0.95 - 0.82 (m, 6H). 13C NMR (101MHz, CDCl3 and 20μL TFA)δ=176.59,176.49,168.80,167.86,159.01,129.77,129.59,122.18,120.85,115.54,73.09,68.74,63.63,63.54 ,63.19,54.21,54.15,34.09,31.77,31.47,29.29,29.16,28.93,25.52,24.67,22.59,13.95,13.87.HRMS(ESI):calcd for C 29 H 48 NO 11 P[M+H] + 618.3038, found 618.3043.

[0344] Example 7 Synthesis of Compound A7

[0345] ethyl 4-(tetradecyloxy)benzoate 1a-1

[0346]

[0347] Ethyl 4-hydroxybenzoate (2.00 g, 12.04 mmol, 1.00 eq.) and 1-bromotetradecane (6.65 g, 24.07 mmol, 2.00 eq.) were dissolved in anhydrous DMF (30 mL). K2CO3 (4.99 g, 36.11 mmol, 3.00 eq.) and KI (0.10 g, 0.60 mmol, 0.05 eq.) were added and stirred at 90°C-100°C for 4.5 h. The reaction was monitored by TLC. After completion, the reaction solution was diluted with water (100 mL) and extracted with dichloromethane (50 mL x 3). The organic layers were combined, washed with 1N dilute hydrochloric acid (50 mL) and then with 1N sodium bicarbonate solution (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA=30 / 1 to 10 / 1) to obtain compound 1a-1 as a white solid in an 86% yield. 1H NMR (400 MHz, cdcl3) δ 8.02 - 7.94 (m, 2H), 6.93 - 6.86 (m, 2H), 4.34 (q, J = 7.1 Hz, 2H), 3.99 (t, J = 6.6 Hz, 2H), 1.79 (dt, J = 14.7, 6.7 Hz, 2H), 1.45 (p, J = 6.7 Hz, 2H), 1.37 (t, J = 7.1 Hz, 3H), 1.26 (s, 20H), 0.92 - 0.84 (m, 3H). HRMS (ESI): calcd for C 23 H 38 O3[M+H] + 363.2894, found 363.2907.

[0348] 4-(tetradecyloxy)benzoic acid 1a-2

[0349]

[0350] NaOH (2.81 g, 70.21 mmol, 5.00 eq.) was dissolved in water (15 mL) and added to a solution of 1a-1 (5.09 g, 14.04 mmol, 1.00 eq.) in ethanol (30 mL) and stirred at 90 °C overnight. The reaction was monitored by TLC and upon completion, the reaction was diluted with water (50 mL) and the pH was adjusted to 2 using 2N dilute hydrochloric acid. A large amount of white precipitate was obtained, which was allowed to settle and filtered to obtain a white solid. The compound 1a-2 was obtained as a white solid after drying with a yield of 62%. 1 H NMR (400 MHz, Chloroform-d) δ 8.06 - 8.02 (m, 2H), 6.95 - 6.90 (m, 2H), 4.02 (t, J = 6.6 Hz, 2H), 1.80 (p, J = 6.7 Hz, 2H), 1.45 (q, J = 7.2 Hz, 2H), 1.26 (s, 21H), 0.88 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 21 H 34 O3[M+Na] + 357.24, found 357.2420.

[0351] (S)-3-(trityloxy)propane-1,2-diyl bis(4-(tetradecyloxy)benzoate)3a-7

[0352]

[0353] The compound 1a-2 was treated according to the method described in 3a-1 to give the target compound as colorless transparent oil in 55% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.02 (d, J = 8.5 Hz, 2H), 7.84 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 7.3 Hz, 6H), 7.22 (t, J = 8.1 Hz, 9H), 6.91 (d, J = 8.6 Hz, 2H), 6.84 (d, J = 8.5 Hz, 2H), 5.59 (dt, J = 10.3, 5.2 Hz, 1H), 4.64 (t, J = 4.3 Hz, 2H), 4.00 (q, J = 6.7 Hz, 4H), 3.47 (dd, J = 10.0, 4.6 Hz, 1H), 3.40 (dd, J = 10.0, 4.7 Hz, 1H), 1.83 - 1.75 (m, 4H), 1.51 - 1.20 (m, 44H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 64 H 86 O7[M+Na] + 989.6266, found 989.6261.

[0354] (S)-3-hydroxypropane-1,2-diyl bis(4-(tetradecyloxy)benzoate)4a-7

[0355]

[0356] The compound 1a-2 was treated according to the method described in 3a-1 to give the target compound as colorless transparent oil in 55% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.02 (d, J = 8.5 Hz, 2H), 7.84 (d, J = 8.6 Hz, 2H), 7.45 (d, J = 7.3 Hz, 6H), 7.22 (t, J = 8.1 Hz, 9H), 6.91 (d, J = 8.6 Hz, 2H), 6.84 (d, J = 8.5 Hz, 2H), 5.59 (dt, J = 10.3, 5.2 Hz, 1H), 4.64 (t, J = 4.3 Hz, 2H), 4.00 (q, J = 6.7 Hz, 4H), 3.47 (dd, J = 10.0, 4.6 Hz, 1H), 3.40 (dd, J = 10.0, 4.7 Hz, 1H), 1.83 - 1.75 (m, 4H), 1.51 - 1.20 (m, 44H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 45 H 72 O7[M+Na] + 747.517, found 747.5178.

[0357] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(4-(tetradecyloxy)benzoate) 6a-7

[0358]

[0359] Following the procedure described in 5a-1, 4a-7 was treated to give the target compound as a colorless transparent oil in 28% yield. 1 HNMR (400 MHz, Chloroform-d) δ 7.95 (dt, J = 19.8, 9.7 Hz, 4H), 6.88 (dd, J = 9.1, 2.6 Hz, 4H), 5.50 (d, J = 8.5 Hz, 1 H), 4.71 - 3.83 (m, 1 1 H), 1.78 (q, J = 6.5, 6.0 Hz, 4H), 1.51 - 1.08 (m, 71 H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 61 H 102 NO 13 P[M + Na] + 1110.6981, found 1110.6973.

[0360] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(4-(tetradecyloxy)benzoate) 6a-7

[0361]

[0362] Following the procedure described in 6a-1, 5a-7 was treated to give the target compound as a colorless transparent oil in 85% yield. 1HNMR (400 MHz, Chloroform-d) δ 7.97 (dd, J = 13.9, 8.6 Hz, 4H), 6.89 (dd, J = 8.9, 1.9 Hz, 4H), 5.62 - 5.50 (m, 2H), 4.67 - 4.48 (m, 2H), 4.42 - 4.16 (m, 4H), 3.99 (t, J = 6.5 Hz, 4H), 1.79 (p, J = 6.8 Hz, 4H), 1.51 - 1.39 (m, 29H), 1.26 (s, 42H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 61 H 102 NO 14 P[M+Na] + 1126.693, found 1126.6931.

[0363] O-(((R)-2,3-bis((4-(tetradecyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A7

[0364]

[0365] Following the procedure described for A1, 6a-7 was treated to give the target compound as a light yellow solid in 35% yield. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.91 (br dd, J = 8.8, 15.2 Hz, 4H), 7.75 (br s, 2H), 6.89 (br dd, J = 5.2, 8.3 Hz, 4H), 5.63 (br s, 1H), 4.74 - 4.51 (m, 4H), 4.50 - 4.25 (m, 3H), 4.00 (br t, J = 6.4 Hz, 4H), 1.87 - 1.70 (m, 4H), 1.45 (br s, 4H), 1.27 (s, 40H), 0.89 (br t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 168.80, 168.24, 167.78, 164.17, 164.03, 132.17, 132.06, 120.17, 120.02, 114.49, 70.77, 68.51, 65.23, 63.68, 63.13, 62.75, 54.18, 31.92, 29.65, 29.36, 29.00, 25.91, 22.67, 14.06. HRMS (ESI): calcd for C48 H 78 NO 12 P[M+H] + 892.5334, found 892.5331.

[0366] Synthesis of compound A8 of Example 8

[0367] 3-(tetradecyloxy)benzoic acid 1a-3

[0368]

[0369] KOH (2.24 g, 40.00 mmol, 2.00 eq.) was first dissolved in water (20 mL), and then 3-hydroxybenzoic acid (2.76 g, 20.00 mmol, 1.00 eq.), 1-bromotetradecane (11.05 g, 40.00 mmol, 2.00 eq.) were dissolved in ethanol solution (40 mL) and mixed at 120-140 °C for 10 h. The reaction was monitored by TLC. After the reaction was completed, the reaction solution was diluted with water (50 mL), and then adjusted to pH = 2 with 2N dilute hydrochloric acid. A large amount of white precipitate was precipitated. The precipitate was filtered out and washed with water (50 mL) and n-hexane (50 mL) successively. After drying, compound 1a-3 was obtained as a white solid in a yield of 67%. 1 H NMR (400 MHz, Chloroform-d) δ 7.68 (d, J = 7.7 Hz, 1H), 7.60 (s, 1H), 7.36 (t, J = 7.9 Hz, 1H), 7.14 (dd, J = 8.0, 2.5 Hz, 1H), 4.01 (t, J = 6.5 Hz, 2H), 1.80 (p, J = 6.8 Hz, 2H), 1.45 (d, J = 8.1 Hz, 2H), 1.26 (s, 21H), 0.88 (t, J = 6.7 Hz, 3H). HRMS (ESI): calcd for C 21 H 34 O3[M+Na] + 357.2400, found 357.2410.

[0370] (S)-3-(trityloxy)propane-1,2-diyl bis(3-(tetradecyloxy)benzoate)3a-8

[0371]

[0372] Compound 1a and 1a-3 were treated according to the method described in 3a-1 to obtain the target compound as a colorless transparent oil in a yield of 55%. 1H NMR (400 MHz, Chloroform-d) δ 7.66 (d, J = 7.7 Hz, 1H), 7.58 (d, J = 2.5 Hz, 1H), 7.45 (dd, J = 8.2, 1.5 Hz, 8H), 7.34 (t, J = 7.9 Hz, 1H), 7.23 (dd, J = 10.3, 7.4 Hz, 9H), 7.12 - 7.04 (m, 2H), 5.66 - 5.58 (m, 1H), 4.68 (dd, J = 5.5, 3.1 Hz, 2H), 3.93 (dt, J = 18.5, 6.5 Hz, 4H), 3.45 (ddd, J = 25.6, 9.6, 4.4 Hz, 2H), 1.75 (d, J = 9.9 Hz, 4H), 1.64 - 1.19 (m, 44H), 0.88 (t, J = 6.7 Hz, 7H). HRMS (ESI): calcd for C 64 H 86 O7[M+Na] + 989.6266, found 989.6245.

[0373] (S)-3-hydroxypropane-1,2-diyl bis(3-(tetradecyloxy)benzoate)4a-8

[0374]

[0375] Following the procedure described in 4a-1, 3a-8 was treated to give the target compound in 34% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.71 - 7.47 (m, 4H), 7.32 (td, J = 8.0, 3.6 Hz, 2H), 7.09 (d, J = 8.2 Hz, 2H), 5.47 (p, J = 5.1 Hz, 1H), 4.66 (q, J = 6.8, 6.3 Hz, 2H), 3.94 (q, J = 7.3, 6.9 Hz, 6H), 2.58 (s, 1H), 1.77 (p, J = 7.0 Hz, 4H), 1.26 (s, 44H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 45 H 72 O7[M+Na] + 747.517, found 747.5174.

[0376] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(3-(tetradecyloxy)benzoate) 6a-8

[0377]

[0378] Following the procedure described in 5a-1, 5a-8 was treated to give the target compound as a colorless transparent oil in 49% yield. 1 HNMR (400 MHz, Chloroform-d) δ 7.65 - 7.49 (m, 4H), 7.36 - 7.26 (m, 2H), 7.08 (dt, J = 8.4, 3.3 Hz, 2H), 5.64 - 5.48 (m, 2H), 4.69 (ddd, J = 12.0, 5.6, 3.7 Hz, 1H), 4.61 - 4.49 (m, 1H), 4.30 (ddt, J = 11.6, 8.7, 3.1 Hz, 1H), 4.18 - 3.88 (m, 6H), 1.93 (s, 2H), 1.76 (dqd, J = 12.7, 6.6, 4.2 Hz, 4H), 1.50 - 1.23 (m, 71H), 0.96 - 0.77 (m, 6H). HRMS (ESI): calcd for C 61 H 102 NO 13 P[M+Na] + 1110.6981, found 1110.6987.

[0379] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(3-(tetradecyloxy)benzoate) 6a-8

[0380]

[0381] Following the procedure described in 6a-1, 5a-8 was treated to give the target compound as a colorless transparent oil in 84% yield. 1HNMR (400 MHz, Chloroform-d) δ 7.67 - 7.46 (m, 4H), 7.30 (t, J = 12.4 Hz, 4H), 7.06 (d, J = 8.8 Hz, 1H), 4.59 (ddtd, J = 38.1, 32.0, 11.9, 5.3 Hz, 2H), 4.45 - 4.07 (m, 5H), 3.94 (p, J = 6.5 Hz, 4H), 1.76 (h, J = 7.1 Hz, 4H), 1.54 - 1.08 (m, 72H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 61 H 102 NO 14 P[M+Na] + 1126.693, found 1126.6936.

[0382] O-(((R)-2,3-bis((3-(tetradecyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A8

[0383]

[0384] Following the procedure described for Al, 6a-8 were treated to give the target compound as a light yellow solid in 36% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.75 (br s, 3H), 7.57 (br dd, J = 7.6, 16.8 Hz, 2H), 7.49 (br d, J = 16.4 Hz, 2H), 7.34 (dt, J = 5.2, 8.0 Hz, 2H), 7.16 (br d, J = 2.8 Hz, 2H), 5.67 (br s, 1H), 4.78 - 4.69 (m, 1H), 4.65 - 4.51 (m, 3H), 4.49 - 4.27 (m, 3H), 4.05 - 3.88 (m, 4H), 1.76 (br s, 4H), 1.43 (br s, 4H), 1.27 (s, 40H), 0.89 (br t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 168.73, 168.17, 167.69, 158.98, 158.95, 129.79, 129.50, 122.36, 122.27, 121.22, 115.51, 115.24, 71.07, 68.81, 68.72, 65.17, 63.61, 63.25, 54.25, 31.91, 29.65, 29.35, 29.01, 25.88, 22.67, 14.03. HRMS (ESI): calcd for C 48 H 78 NO 12 P[M+H] + 892.5334, found 892.5328.

[0385] Synthesis of compound A9 of Example 9

[0386] methyl 3-nitro-4-(tetradecyloxy)benzoate 1a-4

[0387]

[0388] Methyl 4-hydroxy-3-nitrobenzoate and 1-bromotetradecane were treated according to the procedure described for 1a-1 to give the target compound as a white solid in 96% yield. 1 H NMR (400 MHz, CDC13) δ 8.47 (d, J = 2.2 Hz, 1H), 8.17 (dd, J = 8.9, 2.2 Hz, 1H), 7.09 (d, J = 8.8 Hz, 1H), 4.15 (t, J = 6.4 Hz, 2H), 3.91 (s, 3H), 1.84 (p, J = 6.7 Hz, 2H), 1.47 (p, J = 7.0 Hz, 2H), 1.25 (d, J = 7.1 Hz, 20H), 0.86 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 22 H 35 NO5[M+Na] + 416.2407, found 416.2401.

[0389] 3-nitro-4-(tetradecyloxy)benzoicacid 1a-5

[0390]

[0391] 1a-4 was treated according to the procedure described for 1a-2 to give the target compound as a white solid in 91% yield.1 H NMR (400 MHz, CDC13) δ 8.56 (d, J = 2.2 Hz, 1H), 8.25 (dd, J = 8.8, 2.2 Hz, 1H), 7.14 (d, J = 8.9 Hz, 1H), 4.19 (t, J = 6.4 Hz, 2H), 1.93 - 1.81 (m, 2H), 1.49 (p, J = 7.0 Hz, 2H), 1.40 - 1.19 (m, 20H), 0.88 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 21 H 33 NO5[M+Na] + 402.2251, found 402.2251.

[0392] (S)-3-(trityloxy)propane-1,2-diyl bis(3-nitro-4-(tetradecyloxy)benzoate)3a-9

[0393]

[0394] Following the procedure described in 3a-1, 1a and 1a-5 were treated to give the target compound as a white solid in 43% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.47 (t, J = 1.9 Hz, 1H), 8.29 (t, J = 1.8 Hz, 1H), 8.21 (dt, J = 8.7, 1.8 Hz, 1H), 8.06 (dt, J = 8.7, 1.9 Hz, 1H), 7.45 - 7.41 (m, 6H), 7.32 - 7.18 (m, 9H), 7.12 (d, J = 8.9 Hz, 1H), 7.06 (d, J = 8.9 Hz, 1H), 5.61 (d, J = 5.6 Hz, 1H), 4.67 (qd, J = 11.9, 5.2 Hz, 2H), 4.16 (q, J = 6.3 Hz, 4H), 3.46 (d, J = 5.1 Hz, 2H), 1.85 (t, J = 7.4 Hz, 4H), 1.47 (q, J = 7.0 Hz, 4H), 1.26 (s, 40H), 0.92 - 0.84 (m, 6H). HRMS (ESI): calcd for C 64 H 84 N2O 11 [M+Na] + 1079.5967, found 1079.5972.

[0395] (S)-3-hydroxypropane-1,2-diyl bis(3-nitro-4-(tetradecyloxy)benzoate)4a-9

[0396]

[0397] Following the procedure described in 4a-1, 3a-9 was treated to give the target compound as a white solid in 83% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.47 (d, J = 2.2 Hz, 1H), 8.43 (d, J = 2.1 Hz, 1H), 8.18 (ddd, J = 16.3, 8.9, 2.2 Hz, 2H), 7.26 (s, 2H), 7.11 (dd, J = 8.9, 5.4 Hz, 2H), 5.47 (q, J = 5.1 Hz, 1H), 4.67 (qd, J = 12.1, 5.0 Hz, 2H), 4.16 (td, J = 6.4, 3.3 Hz, 4H), 3.95 (t, J = 5.8 Hz, 2H), 2.15 (t, J = 6.3 Hz, 1H), 1.85 (p, J = 7.0 Hz, 4H), 1.47 (p, J = 7.2 Hz, 4H), 1.39 - 1.21 (m, 71H), 0.87 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 45 H 70 N2O 11 [M+Na] + 837.4872, found 837.4876.

[0398] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)propane-1,2-diyl bis(3-nitro-4-(tetradecyloxy)benzoate)5a-9

[0399]

[0400] Following the procedure described in 5a-1, 4a-9 was treated to give the target compound as a colorless transparent oil in 47% yield. 1HNMR (400 MHz, Chloroform-d) δ 8.49 - 8.42 (m, 2H), 8.24 - 8.11 (m, 2H), 7.10 (dd, J = 9.0, 5.2 Hz, 2H), 5.56 (s, 1H), 4.71 (dd, J = 12.3, 4.2 Hz, 1H), 4.57 (dd, J = 12.5, 6.8 Hz, 1H), 4.32 (t, J = 10.9 Hz, 1H), 4.20 - 3.95 (m, 7H), 1.84 (q, J = 7.6 Hz, 4H), 1.50 - 1.37 (m, 27H), 1.26 (s, 40H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 61 H 100 N3O 17 P[M + Na] + 1200.6683, found 1200.6684.

[0401] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(3-nitro-4-(tetradecyloxy)benzoate)6a-9

[0402]

[0403] Following the procedure described in 6a-1, 5a-9 was treated to give the target compound as a colorless transparent oil in 45% yield. 1 HNMR (400 MHz, Chloroform-d) δ 8.50 - 8.41 (m, 2H), 8.25 - 8.13 (m, 2H), 7.11 (dd, J = 9.0, 5.6 Hz, 2H), 5.61 (s, 1H), 4.77 - 4.67 (m, 1H), 4.58 (dt, J = 12.1, 5.5 Hz, 1H), 4.43 - 4.20 (m, 5H), 4.16 (q, J = 5.8, 5.2 Hz, 4H), 1.85 (p, J = 6.5, 6.0 Hz, 4H), 1.51 - 1.41 (m, 28H), 1.40 - 1.21 (m, 69H), 0.92 - 0.85 (m, 6H). HRMS (ESI): calcd for C 61 H 100 N3O 18 P[M + Na] +1216.6632, found 1216.6624.

[0404] O-(((R)-2,3-bis((3-nitro-4-(tetradecyloxy)benzoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A9

[0405]

[0406] The compound 6a-9 was treated according to the method described in Al to give the target compound as a white solid in 52% yield. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.61 - 8.39 (m, 2H), 8.30 - 8.12 (m, 2H), 7.85 - 7.55 (m, 2H), 7.15 (br d, J = 4.4 Hz, 2H), 5.68 (br s, 1H), 4.88 - 4.30 (m, 7H), 4.18 (br s, 4H), 1.86 (br s, 4H), 1.48 - 1.45 (m, 2H), 1.27 (br s, 40H), 0.95 - 0.85 (m, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 168.95, 165.48, 164.97, 156.88, 156.72, 139.03, 138.90, 136.05, 135.78, 127.70, 127.60, 120.38, 114.29, 71.11, 70.41, 65.29, 63.77, 63.24, 54.26, 31.91, 29.69, 29.64, 29.57, 29.48, 29.35, 29.21, 28.67, 25.69, 22.67, 14.05. HRMS (ESI): calcd for C 48 H 76 N3O 16 P[M-H] - 980.4890, found 980.4863.

[0407] Synthesis of compound A10 of Example 10

[0408] (E)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acrylic acid 1a-6

[0409]

[0410] The procedure described in 1a-3 was followed to treat (E)-3-(4-hydroxy-3- methoxyphenyl)acrylic acid and 1-bromotetradecane to give the target compound as a white solid in 37% yield. 1 H NMR (400 MHz, dmso) δ 7.51 (d, J = 15.9 Hz, 1H), 7.30 (d, J = 2.0 Hz, 1H), 7.17 (dd, J = 8.4, 2.0 Hz, 1H), 6.95 (d, J = 8.3 Hz, 1H), 6.43 (d, J = 15.9 Hz, 1H), 3.97 (t, J = 6.5 Hz, 2H), 3.80 (s, 3H), 1.70 (p, J = 6.7 Hz, 2H), 1.47 - 1.15 (m, 22H), 0.88 - 0.80 (m, 3H). HRMS (ESI): calcd for C 24 H 38 O4[M+H] + 391.2843, found 391.2840.

[0411] (S)-1-(((E)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acryloyl)oxy)-3-(trityloxy)propan-2-yl(Z)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acrylate 3a-10

[0412]

[0413] The procedure described in 3a-1 was followed to treat 1a and 1a-6 to give the target compound as a colorless transparent oil in 50% yield. 1H NMR (400 MHz, cdcl3) δ 7.68 (d, J = 15.9 Hz, 1H), 7.56 (d, J = 15.9 Hz, 1H), 7.47 - 7.43 (m, 6H), 7.28 (dd, J = 8.4, 6.6 Hz, 6H), 7.25 - 7.19 (m, 3H), 7.11 - 7.00 (m, 4H), 6.85 (dd, J = 8.3, 4.4 Hz, 2H), 6.36 (d, J = 15.9 Hz, 1H), 6.24 (d, J = 15.9 Hz, 1H), 5.50 - 5.40 (m, 1H), 4.55 - 4.45 (m, 2H), 3.89 (d, J = 1.5 Hz, 6H), 3.37 (ddd, J = 29.0, 9.9, 4.8 Hz, 2H), 1.85 (p, J = 7.0 Hz, 4H), 1.44 (q, J = 7.1 Hz, 4H), 1.26 (s, 40H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 70 H 94 O9[M+Na] + 1101.6790, found 1101.6783.

[0414] (S)-3-hydroxy-2-(((Z)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acryloyl)oxy)propyl(E)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acrylate 4a-10

[0415]

[0416] Following the procedure described for 4a-1, 3a-10 was treated to give the target compound as a white solid in 80% yield. 1H NMR (400 MHz, cdcl3) δ 7.67 (dd, J = 15.9, 7.9 Hz, 2H), 7.09 (dt, J = 8.3, 2.3 Hz, 2H), 7.05 (dd, J = 4.3, 2.0 Hz, 2H), 6.85 (dd, J = 8.3, 1.4 Hz, 2H), 6.34 (dd, J = 15.9, 13.0 Hz, 2H), 5.27 (q, J = 5.0 Hz, 1H), 4.50 (d, J = 5.1 Hz, 2H), 4.04 (t, J = 6.9 Hz, 4H), 3.89 (s, 6H), 3.86 (t, J = 5.5 Hz, 2H), 1.85 (p, J = 7.0 Hz, 4H), 1.45 (t, J = 7.7 Hz, 4H), 1.40 - 1.23 (m, 40H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 51 H 80 O9[M + Na] + 859.5695, found 859.5696.

[0417] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(((Z)-3-(3-methoxy-4-(tetradecyloxy)phenyl

[0418] acryloyl)oxy)propyl(E)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acrylate5a-10

[0419]

[0420] Following the procedure described in 5a-1, 4a-10 was treated to give the target compound as a colorless transparent oil in 48% yield. 1HNMR(400MHz,cdcl3)δ7.73–7.61(m,2H),7.11–7.02(m,4H),6.85(dd,J=8.2,2.1Hz,2H),6 .38–6.34(m,1H),6.34–6.30(m,1H),5.61(dd,J=8.2,5.0Hz,1H),5.44(p,J=5.0Hz,1H),4.5 0(ddd,J=12.2,8.0,4.3Hz,1H),4.44–4.20(m,6H),4.15–3.98(m,5H),3.89(s,6H),1.86(q, J=7.2Hz,4H),1.53–1.42(m,28H),1.40–1.21(m,43H),0.91–0.85(m,6H).HRMS(ESI):calcd for C 67 H 110 NO 15 P[M+Na] + 1222.7505, found 1222.7510.

[0421] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(((Z)-3-(3-methoxy-4-(tetradecyloxy)phenyl)

[0422] acryloyl)oxy)propyl(E)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acrylate6a-10

[0423]

[0424] 5a-10 was treated according to the method described in 6a-1 to obtain the target compound as a colorless transparent oil in a yield of 44%. 1HNMR (400 MHz, cdcl3) δ 7.70 - 7.58 (m, 2H), 7.08 - 7.03 (m, 4H), 6.82 (dd, J = 8.2, 2.1 Hz, 2H), 6.33 (dd, J = 3.5, 2.6 Hz, 1H), 6.29 (dd, J = 3.5, 2.6 Hz, 1H), 5.58 (dd, J = 8.2, 5.0 Hz, 1H), 5.41 (p, J = 5.0 Hz, 1H), 4.47 (ddd, J = 12.2, 8.0, 4.3 Hz, 1H), 4.42 - 4.32 (m, 3H), 4.24 (tdd, J = 9.5, 8.3, 4.4 Hz, 3H), 4.14 - 3.93 (m, 5H), 3.86 (s, 6H), 1.83 (q, J = 7.2 Hz, 4H), 1.50 - 1.39 (m, 28H), 1.37 - 1.19 (m, 43H), 0.88 - 0.82 (m, 6H). HRMS (ESI): calcd for C 67 H 110 NO 16 P[M+Na] + 1238.7454, found 1238.7463.

[0425] O-(hydroxy((R)-3-(((E)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acryloyl)oxy)-2-(((Z)-3-(3-methoxy-4-(tetradecyloxy)phenyl)acryloyl)oxy)propoxy)phosphoryl)-D-serine A10

[0426]

[0427] Following the procedure described for Al, 6a-10 was treated to give the target compound as a pink solid in 55% yield. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.84 - 7.51 (m, 4H), 7.20 - 7.06 (m, 2H), 6.91 (br dd, J = 2.6, 8.4 Hz, 1H), 6.87 - 6.64 (m, 2H), 6.48 - 6.20 (m, 2H), 5.59 - 5.30 (m, 1H), 4.75 - 4.41 (m, 4H), 4.37 - 4.13 (m, 3H), 4.11 - 4.02 (m, 3H), 3.97 - 3.90 (m, 4H), 3.89 - 3.76 (m, 4H), 1.89 - 1.65 (m, 4H), 1.47 - 1.40 (m, 2H), 1.32 - 1.20 (m, 42H), 0.88 (t, J = 6.6 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 169.35, 168.95, 151.34, 151.24, 148.85, 148.64, 148.20, 147.88, 147.86, 147.38, 126.46, 123.97, 113.25, 112.29, 110.62, 70.66, 69.62, 69.57, 69.33, 65.01, 63.76, 56.25, 55.96, 54.19, 31.93, 29.67, 29.54, 29.37, 28.97, 28.85, 25.86, 22.69, 14.08. HRMS (ESI): calcd for C 54 H 86 NO 14 P[M-H] - 1002.5713, found 1002.5701.

[0428] Synthesis of compound A11 of Example 11

[0429] (S)-3-(trityloxy)propane-1,2-diylbis(2-(((dodecylthio)carbonothioyl)thio)propanoate) 3a-11

[0430]

[0431] Following the procedure described in 3a-1, 1a and 2-(((dodecylthio)thiocarbonoyl)thio)propanoic acid were treated to give the target compound as a brown oil in 71% yield. 1H NMR (400 MHz, cdcl3) δ 7.45 - 7.38 (m, 6 H), 7.31 (t, J = 7.5 Hz, 6 H), 7.25 (d, J = 7.0 Hz, 3 H), 5.26 (h, J = 4.7 Hz, 1 H), 4.87 (tt, J = 7.3, 4.4 Hz, 1 H), 4.80 - 4.70 (m, 1 H), 4.41 (ddt, J = 12.0, 8.1, 3.8 Hz, 1 H), 4.28 (ddt, J = 12.0, 8.7, 6.6 Hz, 1 H), 3.30 (ddt, J = 16.3, 6.9, 3.2 Hz, 6 H), 1.72 - 1.59 (m, 7 H), 1.52 (dd, J = 7.4, 3.6 Hz, 3 H), 1.25 (s, 36 H), 0.88 (t, J = 6.7 Hz, 6 H). HRMS (ESI): calcd for C 54 H 78 O5S6[M+Na] + 1021.4066, found 1021.4064.

[0432] (S)-3-hydroxypropane-1,2-diylbis(2-(((dodecylthio)carbonothioyl)thio)propanoate)4a-11

[0433]

[0434] Following the procedure described in 4a-1, 3a-11 was treated to give the target compound as yellow oil in 71 % yield. 1 H NMR (400 MHz, cdcl3) δ 5.15 - 5.06 (m, 1 H), 4.88 - 4.75 (m, 2 H), 4.35 (dtd, J = 54.6, 1 1.9, 5.0 Hz, 2 H), 3.74 (d, J = 6.5 Hz, 2 H), 3.35 (td, J = 7.2, 2.7 Hz, 4 H), 1.68 (p, J = 7.4 Hz, 4 H), 1.65 - 1.59 (m, 6 H), 1.45 - 1.34 (m, 4 H), 1.25 (s, 32 H), 0.87 (t, J = 6.7 Hz, 6 H), 1.33 - 1.22 (m, 33 H). HRMS (ESI): calcd for C 35 H 64 O5S6[M+Na] + 779.297, found 779.2962.

[0435] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(2-(((dodecylthio)carbonothioyl)thio)propanoate) 5a-11

[0436]

[0437] Following the procedure described in 5a-1, 4a-11 was treated to give the target compound as a yellow oil in 76% yield. 1 H NMR (400 MHz, CDC13) δ 5.20 (s, 1H), 4.90 - 4.75 (m, 2H), 4.44 (dt, J = 12.1, 3.5 Hz, 1H), 4.34 - 4.14 (m, 2H), 4.11 - 4.02 (m, 1H), 4.00 - 3.79 (m, 2H), 3.34 (tt, J = 9.7, 4.9 Hz, 4H), 1.69 (p, J = 7.5 Hz, 4H), 1.60 (d, J = 7.5 Hz, 6H), 1.49 - 1.35 (m, 30H), 1.26 (d, J = 5.4 Hz, 33H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 51 H 94 NO 11 PS6 [M+Na] + 1142.4781, found 1142.4788.

[0438] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)propane-1,2-diyl bis(2-(((dodecylthio)carbonothioyl)thio)propanoate) 5a-11

[0439]

[0440] Following the procedure described in 5a-1, 4a-11 was treated to give the target compound as a yellow oil in 76% yield. 1H NMR (400 MHz, cdcl3) δ 5.26 (s, 1H), 4.90 - 4.75 (m, 2H), 4.52 - 4.29 (m, 3H), 4.27 - 4.05 (m, 4H), 3.34 (tt, J = 7.0, 5.5 Hz, 4H), 1.69 (p, J = 7.4 Hz, 4H), 1.64 - 1.56 (m, 6H), 1.52 - 1.44 (m, 27H), 1.39 (p, J = 6.9 Hz, 2H), 1.26 (d, J = 5.2 Hz, 34H), 0.88 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 51 H 94 NO 12 PS6[M+Na] + 1158.473, found 1158.4721.

[0441] O-(((2R)-2,3-bis((2-(((dodecylthio)carbonothioyl)thio)propanoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine A11

[0442]

[0443] The reaction solution was dissolved in dichloromethane (3.00 mL x 3) and concentrated under reduced pressure to remove excess trifluoroacetic acid. The residue was dissolved in acetonitrile (10.00 mL x 3), ultrasonically shaken and precipitated, and the supernatant was carefully removed with a pipette. The pump was dried, and the target compound was obtained as a yellow oil in a yield of 73%. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.70 (br s, 3H), 5.39 - 5.23 (m, 1H), 4.90 - 4.74 (m, 2H), 4.69 - 4.41 (m, 4H), 4.37 - 4.07 (m, 3H), 3.47 - 3.23 (m, 4H), 1.69 (quin, J = 7.2 Hz, 4H), 1.60 (br d, J = 6.4 Hz, 6H), 1.39 (br d, J = 7.2 Hz, 4H), 1.27 (s, 32H), 0.89 (br t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 221.82, 172.98, 172.76, 168.90, 71.39, 64.71, 63.46, 47.94, 47.82, 47.62, 37.53, 31.91, 29.63, 29.56, 29.34, 29.11, 28.93, 27.78, 27.75, 27.52, 22.67, 16.08, 14.06. HRMS (ESI): calcd for C 38 H 70 NO 10 PS6[M-H] - 922.2989, found 922.2997.

[0444] Synthesis of compound A12 of Example 12

[0445] (S)-2-hydroxy-3-(trityloxy)propyl 4-(tetradecyloxy)benzoate 2a-4

[0446]

[0447] Following the procedure described in 2a-1, 1a-2 and 1a were treated to give the target compound as colorless transparent oil in 64% yield. 1 H NMR (400 MHz, cdcl3) δ 7.89 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 7.6 Hz, 6H), 7.26 (dt, J = 23.3, 7.3 Hz, 9H), 6.88 (d, J = 8.5 Hz, 2H), 4.41 (t, J = 4.6 Hz, 2H), 4.16 - 4.08 (m, 1H), 4.01 (t, J = 6.6 Hz, 2H), 3.31 (d, J = 5.2 Hz, 2H), 2.53 (d, J = 5.4 Hz, 1H), 1.80 (p, J = 6.4 Hz, 2H), 1.46 (p, J = 6.9 Hz, 2H), 1.26 (s, 20H), 0.88 (t, J = 6.5 Hz, 3H). HRMS (ESI): calcd for C 43 H 54 O5[M+Na] + 673.3863, found 673.3873.

[0448] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 4-(tetradecyloxy)benzoate 3a-12

[0449]

[0450] Stearic acid and 2a-4 were treated according to the procedure described in 3a-4 to give the target compound as a white solid in 84% yield. 1 H NMR (400 MHz, CDC13) δ 7.81 - 7.74 (m, 2H), 7.41 - 7.31 (m, 5H), 7.26 - 7.11 (m, 10H), 6.83 - 6.73 (m, 2H), 5.37 - 5.29 (m, 1H), 4.50 - 4.36 (m, 2H), 3.92 (t, J = 6.6 Hz, 2H), 3.25 (qd, J = 9.9, 5.0 Hz, 2H), 2.26 (t, J = 7.5 Hz, 2H), 1.72 (p, J = 6.7 Hz, 2H), 1.54 (d, J = 7.3 Hz, 2H), 1.39 (p, J = 6.8 Hz, 2H), 1.32 - 1.11 (m, 50H), 0.80 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 61 H 88 O6[M+Na] + 939.6473, found 939.6471.

[0451] (S)-3-hydroxy-2-(stearoyloxy)propyl 4-(tetradecyloxy)benzoate 4a-12

[0452]

[0453] 3a-12 was treated according to the procedure described in 4a-1 to give the target compound as a white solid in 87% yield. 1 H NMR (400 MHz, CDC13) δ 8.02 - 7.93 (m, 2H), 6.95 - 6.86 (m, 2H), 4.45 - 4.33 (m, 2H), 4.25 (dt, J = 16.3, 7.4 Hz, 3H), 4.01 (t, J = 6.6 Hz, 2H), 2.62 (d, J = 4.5 Hz, 1H), 2.35 (t, J = 7.6 Hz, 2H), 1.83 - 1.75 (m, 2H), 1.69 - 1.55 (m, 2H), 1.45 (q, J = 7.1 Hz, 2H), 1.26 (d, J = 3.6 Hz, 48H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 42 H 74 O6[M+H] +675.5558, found 675.5548.

[0454] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 4-(tetradecyloxy)benzoate 5a-12

[0455]

[0456] According to the method described in 5a-1, 4a-12 was treated to obtain the target compound, colorless transparent oil, yield 79%. 1 HNMR (400 MHz, cdcl3) δ 7.99 (dd, J = 8.9, 3.1 Hz, 2H), 6.90 (dd, J = 8.9, 2.9 Hz, 2H), 5.66 - 5.51 (m, 1H), 4.71 - 4.50 (m, 1H), 4.45 - 3.95 (m, 9H), 2.32 (td, J = 7.8, 2.7 Hz, 2H), 1.80 (p, J = 6.9 Hz, 2H), 1.65 - 1.55 (m, 2H), 1.50 - 1.22 (m, 77H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 58 H 104 NO 12 P[M+Na] + 1060.7188, found 1060.7183.

[0457] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 4-(tetradecyloxy)benzoate 6a-12

[0458]

[0459] According to the method described in 6a-1, 5a-12 was treated to obtain the target compound, colorless transparent oil, yield 61%. 1HNMR (400 MHz, cdcl3) δ 7.98 (d, J = 8.4 Hz, 2H), 6.90 (dd, J = 8.8, 3.1 Hz, 2H), 5.57 (d, J = 8.1 Hz, 1H), 4.83 (d, J = 10.2 Hz, 1H), 4.54 - 4.19 (m, 7H), 3.99 (t, J = 6.6 Hz, 2H), 2.33 (q, J = 7.9 Hz, 2H), 1.79 (p, J = 6.8 Hz, 2H), 1.61 (dt, J = 14.6, 7.2 Hz, 2H), 1.54 - 1.17 (m, 77H), 0.87 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 58 H 104 NO 13 P[M+Na] + 1076.7137, found 1076.7133.

[0460] O-(hydroxy((R)-2-(stearoyloxy)-3-((4-(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serine A12

[0461]

[0462] According to the method described in A1, 6a-12 was treated to obtain the target compound, white solid, yield 49%. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.93 (d, J = 8.8 Hz, 2H), 7.70 (br s, 2H), 6.93 (d, J = 8.8 Hz, 2H), 4.82 (td, J = 4.5, 9.0 Hz, 1H), 4.60 (br d, J = 3.9 Hz, 3H), 4.53 - 4.41 (m, 3H), 4.35 (br dd, J = 4.8, 11.9 Hz, 1H), 4.02 (t, J = 6.5 Hz, 2H), 2.41 (br t, J = 7.7 Hz, 2H), 1.86 - 1.76 (m, 2H), 1.64 - 1.54 (m, 2H), 1.51 - 1.41 (m, 2H), 1.38 - 1.21 (m, 48H), 0.89 (t, J = 6.6 Hz, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 177.02, 168.85, 168.38, 164.24, 132.12, 119.95, 114.60, 73.33, 68.61, 63.64, 63.39, 54.28, 54.23, 34.14, 31.93, 29.70, 29.60, 29.36, 29.09, 28.98, 25.90, 24.69, 22.68, 14.03. HRMS (ESI): calcd for C 45 H 80 NO 11 P[M-H] - 840.5396, found 840.5425.

[0463] Synthesis of compound A13 of Example 13

[0464] (S)-2-hydroxy-3-(trityloxy)propyl 3-(tetradecyloxy)benzoate 2a-5

[0465]

[0466] Following the procedure described in 2a-1, 1a-3 and 1a were treated to give the target compound as a colorless transparent oil in 64% yield. 1 H NMR (400 MHz, CDC13) δ 7.54 - 7.47 (m, 2H), 7.44 (d, J = 7.3 Hz, 6H), 7.34 - 7.20 (m, 10H), 7.09 (dd, J = 8.1, 2.6 Hz, 1H), 4.51 - 4.36 (m, 2H), 4.13 (s, 1H), 3.96 (t, J = 6.6 Hz, 2H), 3.32 (d, J = 5.2 Hz, 2H), 1.79 (p, J = 6.7 Hz, 2H), 1.45 (d, J = 8.2 Hz, 2H), 1.26 (s, 20H), 0.88 (t, J = 6.7 Hz, 3H). HRMS (ESI): calcd for C 43 H 54 O5[M+Na] + 673.3863, found 673.3841.

[0467] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 3-(tetradecyloxy)benzoate 3a-13

[0468]

[0469] Stearic acid and 2a-5 were treated according to the procedure described in 3a-4 to give the target compound as a colorless transparent oil in 73% yield. 1 H NMR (400 MHz, cdcl3) δ 7.45 (dd, J = 13.4, 8.1 Hz, 8H), 7.29 (t, J = 7.4 Hz, 6H), 7.23 (t, J = 7.0 Hz, 3H), 7.08 (dd, J = 8.2, 2.6 Hz, 1H), 5.41 (p, J = 5.1 Hz, 1H), 4.57 - 4.46 (m, 2H), 3.96 (t, J = 6.5 Hz, 2H), 3.32 (qd, J = 9.8, 4.9 Hz, 2H), 2.34 (t, J = 7.5 Hz, 2H), 1.79 (p, J = 6.8 Hz, 2H), 1.63 - 1.57 (m, 3H), 1.45 (q, J = 7.0 Hz, 2H), 1.36 - 1.19 (m, 48H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 61 H 88 O6[M+Na] + 939.6473, found 939.6469.

[0470] (S)-3-hydroxy-2-(stearoyloxy)propyl 3-(tetradecyloxy)benzoate 4a-13

[0471]

[0472] 3a-13 was treated according to the procedure described in 4a-1 to give the target compound as a white solid in 46% yield. 1 H NMR (400 MHz, cdcl3) δ 7.61 (d, J = 7.7 Hz, 1H), 7.54 (t, J = 2.1 Hz, 1H), 7.34 (t, J = 8.0 Hz, 1H), 7.10 (dd, J = 8.0, 2.6 Hz, 1H), 4.45 - 4.37 (m, 2H), 4.31 - 4.19 (m, 3H), 3.99 (t, J = 6.5 Hz, 2H), 2.39 - 2.28 (m, 2H), 1.84 - 1.74 (m, 2H), 1.62 (p, J = 7.4 Hz, 3H), 1.46 (p, J = 6.9 Hz, 2H), 1.36 - 1.18 (m, 48H), 0.87 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 42 H 74 O6[M+Na] +697.5378, found 697.5369.

[0473] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 3-(tetradecyloxy)benzoate 5a-13

[0474]

[0475] According to the method described in 5a-1, 4a-13 was treated to obtain the target compound, colorless transparent oil, yield 47%. 1 HNMR (400 MHz, CDC13) δ 7.65 - 7.50 (m, 2H), 7.32 (tt, J = 7.6, 2.4 Hz, 1H), 7.09 (dd, J = 8.3, 3.0 Hz, 1H), 5.54 (d, J = 8.5 Hz, 1H), 4.71 - 3.66 (m, 9H), 2.32 (td, J = 7.5, 2.9 Hz, 2H), 1.83 - 1.72 (m, 2H), 1.66 - 1.54 (m, 2H), 1.53 - 1.11 (m, 76H), 0.88 (td, J = 6.8, 2.7 Hz, 6H). HRMS (ESI): calcd for C 58 H 104 NO 12 P[M + Na] + 1060.7188, found 1060.7198.

[0476] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-(tetradecyloxy)benzoate 6a-13

[0477]

[0478] According to the method described in 6a-1, 5a-13 was treated to obtain the target compound, colorless transparent oil, yield 97%. 1HNMR (400 MHz, cdcl3) δ 7.62 (d, J = 7.7 Hz, 1H), 7.56 (t, J = 2.1 Hz, 1H), 7.33 (td, J = 8.0, 3.8 Hz, 1H), 7.09 (dd, J = 8.2, 2.7 Hz, 1H), 5.56 (t, J = 8.1 Hz, 1H), 4.83 (dp, J = 8.9, 4.6 Hz, 1H), 4.56 - 4.21 (m, 7H), 3.99 (t, J = 6.5 Hz, 2H), 2.33 (q, J = 8.0 Hz, 2H), 1.78 (p, J = 6.8 Hz, 2H), 1.64 - 1.56 (m, 2H), 1.51 - 1.40 (m, 29H), 1.25 (q, J = 5.0 Hz, 48H), 0.87 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 58 H 104 NO 13 P[M+Na] + 1076.7137, found 1076.7131.

[0479] O-(hydroxy((R)-2-(stearoyloxy)-3-((3-(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serine A13

[0480]

[0481] Following the procedure described for Al, 6a-13 was treated to give the target compound as a light yellow solid in 57% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.73 (br s, 3H), 7.58 (br d, J = 7.6 Hz, 1H), 7.51 (s, 1H), 7.35 (t, J = 8.0 Hz, 1H), 7.16 (dd, J = 2.0, 8.2 Hz, 1H), 4.86 - 4.74 (m, 1H), 4.66 - 4.54 (m, 3H), 4.53 - 4.38 (m, 3H), 4.33 (br dd, J = 4.8, 12.0 Hz, 1H), 4.00 (t, J = 6.8 Hz, 2H), 2.39 (br t, J = 7.6 Hz, 2H), 1.85 - 1.73 (m, 2H), 1.64 - 1.54 (m, 2H), 1.48 - 1.42 (m, 2H), 1.34 - 1.23 (m, 48H), 0.89 (t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.77, 168.84, 167.99, 159.00, 129.79, 129.53, 122.24, 120.97, 115.54, 73.16, 68.79, 63.63, 63.26, 54.26, 54.20, 34.12, 31.92, 29.70, 29.57, 29.36, 29.10, 29.02, 28.97, 25.88, 24.69, 22.67, 14.05. HRMS (ESI): calcd for C 45 H 80 NO 11 P[M+H] + 842.5542, found 842.5540.

[0482] Synthesis of compound A14 of Example 14

[0483] ethyl 2-(tetradecyloxy)benzoate 1a-7

[0484]

[0485] Following the procedure described in 1a-1, ethyl 2-hydroxybenzoate and 1- bromotetradecane were treated to give the target compound as a colorless transparent oil in 81% yield. 1 H NMR (400 MHz, CDC13) δ 7.77 (d, J = 7.7 Hz, 1H), 7.48 - 7.38 (m, 1H), 6.96 (t, J = 7.7 Hz, 2H), 4.36 (q, J = 7.2 Hz, 2H), 4.02 (t, J = 6.5 Hz, 2H), 1.83 (p, J = 6.8 Hz, 2H), 1.49 (dd, J = 14.4, 8.0 Hz, 2H), 1.38 (t, J = 7.2 Hz, 3H), 1.26 (s, 20H), 0.87 (d, J = 7.0 Hz, 3H). HRMS (ESI): calcd for C 23 H 38 O3[M+H] + 363.2894, found 363.2894.

[0486] 2-(tetradecyloxy)benzoic acid 1a-8

[0487]

[0488] Treatment of 1a-7 according to the procedure described in 1a-2 gave the target compound as a white solid in 78% yield. 1 H NMR (400 MHz, dmso) δ 7.60 (dd, J = 7.6, 1.8 Hz, 1H), 7.46 (ddd, J = 8.4, 7.4, 1.9 Hz, 1H), 7.09 (dd, J = 8.5, 1.0 Hz, 1H), 6.97 (td, J = 7.5, 1.0 Hz, 1H), 4.01 (t, J = 6.4 Hz, 2H), 1.76 - 1.63 (m, 2H), 1.42 (p, J = 6.9 Hz, 2H), 1.24 (s, 20H), 0.89 - 0.79 (m, 3H). HRMS (ESI): calcd for C 21 H 34 O3[M+H] + 335.2581, found 335.2578.

[0489] (S)-2-hydroxy-3-(trityloxy)propyl 2-(tetradecyloxy)benzoate 2a-6

[0490]

[0491] Treatment of 1a-8 and 1a according to the procedure described in 2a-1 gave the target compound as a colorless transparent oil in 75% yield. 1 H NMR (400 MHz, cdcl3) δ 7.72 (dt, J = 7.9, 1.6 Hz, 1H), 7.44 (d, J = 8.2 Hz, 6H), 7.32 - 7.17 (m, 9H), 6.94 (dt, J = 7.5, 3.2 Hz, 2H), 4.47 (dd, J = 11.4, 4.2 Hz, 1H), 4.43 - 4.36 (m, 1H), 4.12 (q, J = 5.4 Hz, 1H), 3.98 (t, J = 6.8 Hz, 2H), 3.37 - 3.24 (m, 2H), 1.76 (p, J = 7.0 Hz, 2H), 1.41 (d, J = 10.1 Hz, 2H), 1.25 (s, 20H), 0.88 (t, J = 6.7 Hz, 3H). HRMS (ESI): calcd for C 43 H 54 O5[M+Na] + 673.3863, found 673.3860.

[0492] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 2-(tetradecyloxy)benzoate 3a-14

[0493]

[0494] Stearic acid and 2a-6 were treated according to the procedure described in 3a-4 to give the target compound as a white solid in 90% yield. 1 H NMR (400 MHz, cdcl3) δ 7.64 (dd, J = 7.6, 1.9 Hz, 1H), 7.46 - 7.38 (m, 6H), 7.31 - 7.17 (m, 9H), 6.91 (dd, J = 13.7, 7.6 Hz, 2H), 5.38 (q, J = 5.2 Hz, 1H), 4.51 (h, J = 6.9 Hz, 2H), 3.96 (t, J = 6.8 Hz, 2H), 3.32 (qd, J = 9.9, 5.0 Hz, 2H), 2.33 (t, J = 7.6 Hz, 2H), 1.76 (p, J = 7.0 Hz, 2H), 1.62 (q, J = 7.4 Hz, 2H), 1.42 (p, J = 7.2 Hz, 2H), 1.36 - 1.15 (m, 48H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 61 H 88 O6[M+Na] + 939.6473, found 939.6487.

[0495] (S)-3-hydroxy-2-(stearoyloxy)propyl 2-(tetradecyloxy)benzoate 4a-14

[0496]

[0497] 3a-14 was treated according to the procedure described in 4a-1 to give the target compound as a white solid in 74% yield. 1H NMR (400 MHz, cdcl3) δ 7.83 - 7.78 (m, 1H), 7.46 (t, J = 7.9 Hz, 1H), 6.98 (t, J = 7.6 Hz, 2H), 4.42 (dd, J = 11.6, 3.9 Hz, 1H), 4.35 (dd, J = 11.5, 5.6 Hz, 1H), 4.27 - 4.16 (m, 3H), 4.04 (t, J = 6.9 Hz, 2H), 2.72 (d, J = 4.7 Hz, 1H), 2.35 (t, J = 7.6 Hz, 2H), 1.83 (p, J = 7.0 Hz, 2H), 1.63 (p, J = 6.9 Hz, 2H), 1.45 (p, J = 7.1 Hz, 2H), 1.25 (d, J = 2.6 Hz, 48H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 42 H 74 O6[M+Na] + 697.5378, found 697.5378.

[0498] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 2-(tetradecyloxy)benzoate 5a-14

[0499]

[0500] Following the procedure described for 5a-1, 4a-14 was treated to give the target compound as a colorless transparent oil in 45% yield. 1 H NMR (400 MHz, cdcl3) δ 7.80 (t, J = 6.9 Hz, 1H), 7.43 (t, J = 7.9 Hz, 1H), 6.94 (d, J = 8.2 Hz, 2H), 5.56 (q, J = 9.7 Hz, 1H), 4.60 (d, J = 20.3 Hz, 1H), 4.44 - 4.06 (m, 7H), 4.02 (t, J = 6.7 Hz, 2H), 2.33 (q, J = 7.3 Hz, 2H), 1.82 (p, J = 7.1 Hz, 2H), 1.57 (s, 2H), 1.47 - 1.20 (m, 75H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 58 H 104 NO 12 P[M+Na] +1060.7188, found 1060.7185.

[0501] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 2-(tetradecyloxy)benzoate 6a-14

[0502]

[0503] Following the procedure described in 6a-1, 5a-14 was treated to give the target compound as a colorless transparent oil in 61% yield. 1 HNMR (400 MHz, cdcl3) δ 7.82 (d, J = 7.8 Hz, 1H), 7.44 (t, J = 7.9 Hz, 1H), 6.95 (d, J = 8.1 Hz, 2H), 5.65 - 5.50 (m, 1H), 4.80 (dd, J = 9.1, 4.9 Hz, 1H), 4.51 - 4.15 (m, 7H), 4.02 (t, J = 6.9 Hz, 2H), 2.36 - 2.29 (m, 2H), 1.83 (p, J = 7.1 Hz, 2H), 1.60 (d, J = 7.4 Hz, 2H), 1.50 - 1.19 (m, 77H), 0.87 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 58 H 104 NO 13 P[M+Na] + 1076.7137, found 1076.7140.

[0504] O-(hydroxy((R)-2-(stearoyloxy)-3-((2-(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serine A14

[0505]

[0506] Following the procedure described in A1, 6a-14 was treated to give the target compound as a white solid in 30% yield. 1H NMR (400 MHz, CDC13 and 20 uL TFA) δ = 7.83 (br d, J = 7.2 Hz, 1H), 7.69 (br s, 2H), 7.54 (br t, J = 7.2 Hz, 1H), 7.05 - 6.93 (m, 2H), 4.92 - 4.78 (m, 1H), 4.69 - 4.22 (m, 7H), 4.06 (br t, J = 6.8 Hz, 2H), 2.42 (br t, J = 7.6 Hz, 2H), 1.86 - 1.73 (m, 2H), 1.61 (br s, 2H), 1.40 (br s, 2H), 1.27 (br s, 48H), 0.89 (br t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 uL TFA) δ = 176.86, 167.96, 159.29, 135.43, 132.08, 120.34, 117.25, 113.38, 73.29, 73.24, 69.28, 63.57, 63.43, 54.26, 34.12, 31.92, 29.70, 29.65, 29.57, 29.42, 29.36, 29.26, 29.10, 28.64, 25.69, 22.67, 14.05. HRMS (ESI): calcd for C 45 H 80 NO 11 P[2M-H] - 1683.0898, found 1683.0881.

[0507] Synthesis of compound A15 of Example 15

[0508] 3-bromo-4-pentadecylbenzoic acid 1a-9

[0509]

[0510] To a solution of 3-bromo-4-methyl-benzoic acid (8.00 g, 37.2 mmol, 1.00 eq.) in tetrahydrofuran (80 mL) was added lithium diisopropylamide (2.00 M, 40.90 mL, 2.20 eq.) at -40 °C. The reaction was warmed to 25 °C and stirred for 2 h. Then a solution of 1-bromotetradecane (11.40 g, 40.90 mmol, 12.20 mL, 1.10 eq.) in tetrahydrofuran (40.0 mL) was added to the mixture. The mixture was stirred at 25 °C for 8 h. The mixture was quenched with hydrochloric acid (50.00 mL, 2.00 M) and extracted with ethyl acetate (100 mL x 3). The organic phase was combined, washed with brine (50.00 mL), dried, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (PE / EA = 20 / 1 ~ 5 / 1) to give compound 1a-9 as a white solid in 41% yield. 1 HNMR (400 MHz, CDC13) δ = 8.27 (d, J = 1.6 Hz, 1H), 7.97 (d, J = 8.0 Hz, 1H), 7.33 (dd, J = 8.0 Hz, 1H), 2.82-2.78 (m, 2H), 1.67-1.66 (m, 2H), 1.60-1.62 (m, 2H), 1.27 (m, 22H), 0.87-0.91 (m, 3H). HRMS (ESI): calcd for C 22 H 35 BrO2[M+H] + 411.1893, found 411.1902.

[0511] (S)-2-hydroxy-3-(trityloxy)propyl 3-bromo-4-pentadecylbenzoate 2a-7

[0512]

[0513] According to the method described in 2a-1, 1a-9 and 1a were treated to give the target compound as a yellow oil in 40% yield. 1HNMR (400 MHz, CDC13) δ = 8.12 (d, J = 1.6 Hz, 1H), 7.80 (dd, J = 1.6, 8.0 Hz, 1H), 7.46-7.44 (m, 6H), 7.32-7.30 (m, 6H), 7.28-7.27 (m, 4H), 4.50-4.38 (m, 2H), 4.15-4.11 (m, 1H), 3.33 (dd, J = 2.0, 5.2 Hz, 2H), 2.84-2.73 (m, 2H), 2.45 (d, J = 5.6 Hz, 1H), 1.67-1.58 (m, 2H), 1.35 (br s, 2H), 1.32-1.23 (m, 22H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 44 H 55 BrO4[M+Na] + 749.3176, found 749.3182.

[0514] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 3-bromo-4-pentadecylbenzoate3a-15

[0515]

[0516] Stearic acid and 2a-7 were treated according to the procedure described in 3a-4 to give the target compound as a yellow oil in 92% yield. 1 H NMR (400 MHz, CDC13) δ = 8.08 (d, J = 1.6 Hz, 1H), 7.75 (dd, J = 1.6, 8.0 Hz, 1H), 7.45 (d, J = 7.2 Hz, 6H), 7.31-7.29 (m, 5H), 7.28-7.24 (m, 5H), 5.45-5.34 (m, 1H), 4.64-4.43 (m, 2H), 3.36-3.28 (m, 2H), 2.79-2.75 (m, 2H), 2.34 (t, J = 7.2 Hz, 2H), 1.67-1.58 (m, 4H), 1.32-1.21 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 62 H 89 BrO5[M+Na] + 1015.5786, found 1015.5779.

[0517] (S)-3-hydroxy-2-(stearoyloxy)propyl 3-bromo-4-pentadecylbenzoate 4a-15

[0518]

[0519] Following the procedure described in 4a-1, 3a-15 was treated to give the target compound as a brown solid in 50% yield. 1 H NMR (400 MHz, CDC13) δ = 8.40 (d, J = 1.6 Hz, 1H), 7.89 (dd, J = 1.6, 8.0 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 5.24 (quin, J = 5.2 Hz, 1H), 4.52 (dq, J = 5.2, 11.6 Hz, 2H), 3.83 (br d, J = 4.4 Hz, 2H), 2.80-2.76 (m, 2H), 2.38 (t, J = 7.6 Hz, 2H), 2.16 (br d, J = 0.8 Hz, 1H), 1.65-1.61 (m, 4H), 1.28-1.25 (m, 52H), 0.90 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 43 H 75 BrO5[M+Na] + 773.4690, found 773.4672.

[0520] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 3-bromo-4-pentadecylbenzoate 5a-15

[0521]

[0522] Following the procedure described in 5a-1, 4a-15 was treated to give the target compound as a colorless transparent oil in 23% yield. 1HNMR (400 MHz, CDC13) δ = 8.16 (d, J = 1.6 Hz, 1H), 7.86 (dd, J = 1.2, 8.0 Hz, 1H), 7.29-7.27 (m, 1H), 5.49 (br d, J = 7.6 Hz, 1H), 5.37-5.28 (m, 1H), 4.60-4.52 (m, 1H), 4.48-4.37 (m, 1H), 4.37-4.24 (m, 1H), 4.20-3.88 (m, 4H), 2.84-2.71 (m, 2H), 2.33 (t, J = 7.2 Hz, 2H), 1.66-1.60 (m, 4H), 1.46 (d, J = 8.4 Hz, 18H), 1.41 (d, J = 6.8 Hz, 9H), 1.35-1.20 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 105 BrNO 11 P[M+Na] + 1136.6501, found 1136.6511.

[0523] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-bromo-4-pentadecylbenzoate 6a-15

[0524]

[0525] Following the procedure described for 6a-1, 5a-15 was treated to give the target compound as a colorless transparent oil in 62% yield. 1HNMR (400 MHz, CDC13) δ = 8.16 (s, 1H), 7.86 (d, J = 8.0 Hz, 1H), 7.29 (s, 1H), 5.57-5.47 (m, 1H), 5.45-5.34 (m, 1H), 4.62-4.52 (m, 1H), 4.47-4.32 (m, 3H), 4.29-4.15 (m, 3H), 2.83-2.72 (m, 2H), 2.35 (dt, J = 3.6, 7.6 Hz, 2H), 1.61 (td, J = 7.6, 14.6 Hz, 4H), 1.52-1.43 (m, 27H), 1.35 (br s, 4H), 1.30-1.21 (m, 48H), 0.88 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 105 BrNO 12 P[M+Na] + 1152.6450, found 1152.6457.

[0526] O-(((R)-3-((3-bromo-4-pentadecylbenzoyl)oxy)-2-(stearoyloxy)propoxy)(hydroxy)phosphoryl)-D-serine A15

[0527]

[0528] According to the method described in Al, 6a-15 was treated to obtain the target compound, white solid, yield 76%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.12 (s, 1H), 7.82 (br d, J = 8.0 Hz, 1H), 7.70 (br s, 2H), 7.32 (br d, J = 8.0 Hz, 1H), 5.49 (br s, 1H), 4.63 (br d, J = 15.2 Hz, 3H), 4.57-4.43 (m, 2H), 4.25 (br s, 2H), 2.79 (br t, J = 7.6 Hz, 2H), 2.50-2.37 (m, 2H), 1.67-1.55 (m, 4H), 1.37 (br d, J = 5.2 Hz, 4H), 1.32-1.18 (m, 44H), 0.89 (t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 177.09, 168.91, 167.14, 149.57, 134.05, 130.40, 128.59, 127.24, 124.64, 70.63, 70.59, 65.13, 63.60, 63.35, 54.34, 36.50, 31.91, 29.69, 29.65, 29.56, 29.38, 29.35, 29.06, 28.91, 24.81, 22.65, 13.99. HRMS (ESI): calcd for C 46 H 81 BrNO 10 P[M-H] - 916.4709, found 916.4721.

[0529] Synthesis of compound A16 of Example 16

[0530] 3-chloro-4-pentadecylbenzoic acid 1a-10

[0531]

[0532] 3-chloro-4-methylbenzoic acid was treated according to the procedure described for 1a-9 to give the target compound as a colorless transparent oil in 52% yield. 1 H NMR (400 MHz, DMSO-d6) δ = 13.14 (br s, 1H), 7.86 (s, 1H), 7.80 (br d, J = 8.0 Hz, 1H), 7.44 (br d, J = 8.0 Hz, 1H), 2.76-2.67 (m, 2H), 1.55 (br s, 2H), 1.31-1.18 (m, 24H), 0.88-0.80 (m, 3H). HRMS (ESI): calcd for C 22 H 35 ClO2[M+H] + 367.2398, found 367.2403.

[0533] (S)-2-hydroxy-3-(trityloxy)propyl 3-chloro-4-pentadecylbenzoate 2a-8

[0534]

[0535] 1a-10 and 1a were treated according to the procedure described for 2a-1 to give the target compound as a colorless transparent oil in 44% yield.1 HNMR (400 MHz, CDC13) δ = 7.91 (d, J = 1.6 Hz, 1H), 7.77 (dd, J = 1.6, 8.0 Hz, 1H), 7.48-7.42 (m, 6H), 7.30 (br d, J = 7.6 Hz, 7H), 7.27-7.22 (m, 3H), 4.50-4.38 (m, 2H), 4.15-4.11 (m, 1H), 3.33 (dd, J = 2.0, 5.2 Hz, 2H), 2.84-2.73 (m, 2H), 2.45 (d, J = 5.6 Hz, 1H), 1.67-1.58 (m, 2H), 1.35 (br s, 2H), 1.32-1.23 (m, 22H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 44 H 55 ClO4[M+Na] + 705.3681, found 705.3688.

[0536] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 3-chloro-4-pentadecylbenzoate3a-16

[0537]

[0538] Stearic acid and 2a-8 were treated according to the method described in 3a-4 to give the target compound as colorless transparent oil in 90% yield. 1 HNMR (400 MHz, CDC13) δ = 7.87 (d, J = 1.6 Hz, 1H), 7.72 (dd, J = 1.6, 8.0 Hz, 1H), 7.44 (d, J = 7.2 Hz, 6H), 7.32-7.27 (m, 6H), 7.26-7.21 (m, 4H), 5.45-5.34 (m, 1H), 4.64-4.43 (m, 2H), 3.40-3.27 (m, 2H), 2.84-2.71 (m, 2H), 2.34 (t, J = 7.2 Hz, 2H), 1.67-1.58 (m, 4H), 1.32-1.21 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 62 H 89 ClO5[M+Na] + 971.6291, found 971.6303.

[0539] (S)-3-hydroxy-2-(stearoyloxy)propyl 3-chloro-4-pentadecylbenzoate 4a-16

[0540]

[0541] Treatment of 3a-16 according to the procedure described in 4a-1 gave the target compound as a white solid in 65% yield. 1 H NMR (400 MHz, CDC13) δ = 7.98 (d, J = 1.6 Hz, 1H), 7.83 (dd, J = 1.6, 8.0 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 5.24 (quin, J = 5.2 Hz, 1H), 4.52 (dq, J = 5.2, 11.6 Hz, 2H), 3.83 (br d, J = 4.4 Hz, 2H), 2.82 - 2.72 (m, 2H), 2.37 (t, J = 7.6 Hz, 2H), 2.11 (br d, J = 0.8 Hz, 1H), 1.68 - 1.58 (m, 4H), 1.38 - 1.23 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 43 H 75 ClO5[M+Na] + 729.5195, found 729.5187.

[0542] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 3-chloro-4-pentadecylbenzoate 5a-16

[0543]

[0544] Treatment of 4a-16 according to the procedure described in 5a-1 gave the target compound as a colorless transparent oil in 12% yield. 1HNMR (400 MHz, CDC13) δ = 7.98 (d, J = 1.2 Hz, 1H), 7.82 (d, J = 7.6 Hz, 1H), 7.29 (s, 1H), 5.48 (br d, J = 7.6 Hz, 1H), 5.38-5.30 (m, 1H), 4.61-4.51 (m, 1H), 4.46-4.38 (m, 1H), 4.31 (br dd, J = 3.2, 8.4 Hz, 1H), 4.20-3.98 (m, 3H), 3.97-3.89 (m, 1H), 2.80-2.73 (m, 2H), 2.33 (t, J = 7.6 Hz, 2H), 1.66-1.59 (m, 4H), 1.48-1.40 (m, 27H), 1.26 (s, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 105 ClNO 11 P[M+Na] + 1092.7006, found 1092.7022.

[0545] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-chloro-4-pentadecylbenzoate 6a-16

[0546]

[0547] Following the procedure described in 6a-1, 5a-16 was treated to give the target compound as a colorless transparent oil in 66% yield. 1HNMR (400 MHz, CDC13) δ = 7.97 (s, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.29 (d, J = 8.0 Hz, 1H), 5.59-5.48 (m, 1H), 5.43-5.32 (m, 1H), 4.62-4.51 (m, 1H), 4.47-4.33 (m, 3H), 4.29-4.15 (m, 3H), 2.81-2.73 (m, 2H), 2.35 (dt, J = 3.6, 7.6 Hz, 2H), 1.61 (br d, J = 7.6 Hz, 4H), 1.51-1.47 (m, 18H), 1.45 (d, J = 1.6 Hz, 9H), 1.31-1.20 (m, 52H), 0.88 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 105 ClNO 12 P[M+Na] + 1108.6955, found 1108.6971.

[0548] O-(((R)-3-((3-chloro-4-pentadecylbenzoyl)oxy)-2-(stearoyloxy)propoxy)(hydroxy)phosphoryl)-D-serine A16

[0549]

[0550] According to the method described in Al, 6a-16 was treated to obtain the target compound, white solid, yield 78%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.92 (s, 1H), 7.77 (br d, J = 8.0 Hz, 3H), 7.31 (d, J = 8.0 Hz, 1H), 5.46 (br d, J = 2.8 Hz, 1H), 4.61 (br d, J = 9.2 Hz, 3H), 4.48 (br d, J = 6.8 Hz, 2H), 4.28-4.16 (m, 2H), 2.77 (br t, J = 7.6 Hz, 2H), 2.48-2.36 (m, 2H), 1.69-1.55 (m, 4H), 1.35 (br s, 4H), 1.31-1.17 (m, 48H), 0.89 (t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 177.09, 168.81, 167.36, 147.89, 134.59, 130.66, 130.56, 127.96, 127.09, 70.62, 70.53, 65.10, 63.61, 63.33, 54.33, 54.27, 34.33, 33.89, 31.91, 29.69, 29.65, 29.56, 29.38, 29.34, 29.06, 28.91, 24.80, 22.65, 13.96. HRMS (ESI): calcd for C 46 H 81 ClNO 10 P[M-H] - 872.5214, found 872.5205.

[0551] Synthesis of compound A17 of Example 17

[0552] 3-bromo-5-chloro-4-methylbenzoic acid 1a-11

[0553]

[0554] 3-bromo-5-chloro-4-methylbenzoic acid was treated according to the procedure described for 1a-9 to give the target compound as a white solid in 78% yield. 1 H NMR (400 MHz, CDC13) δ = 8.17 (d, J = 1.6 Hz, 1H), 8.02 (d, J = 1.6 Hz, 1H), 3.06-2.94 (m, 2H), 1.63-1.53 (m, 2H), 1.49-1.41 (m, 2H), 1.39-1.21 (m, 22H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C8H6BrCl02[M+H] + 445.1503, found 445.1512.

[0555] (S)-2-hydroxy-3-(trityloxy)propyl 3-bromo-5-chloro-4-pentadecylbenzoate 2a-9

[0556]

[0557] 1a-11 and 1a were treated according to the procedure described for 2a-1 to give the target compound as a yellow oil in 30% yield. 1HNMR (400 MHz, CDC13) δ = 8.03 (d, J = 1.6 Hz, 1H), 7.88 (d, J = 1.6 Hz, 1H), 7.51-7.42 (m, 6H), 7.35-7.24 (m, 9H), 4.44 (d, J = 5.2 Hz, 2H), 4.14-4.10 (m, 1H), 3.40-3.27 (m, 2H), 3.04-2.92 (m, 2H), 2.46 (br d, J = 5.2 Hz, 1H), 1.60-1.54 (m, 2H), 1.51-1.42 (m, 2H), 1.42-1.28 (m, 22H), 0.96-0.84 (m, 3H). HRMS (ESI): calcd for C 44 H 54 BrClO4[M+Na] + 783.2786, found 783.2788.

[0558] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 3-bromo-5-chloro-4-pentadecylbenzoate 3a-17

[0559]

[0560] Stearic acid and 2a-9 were treated according to the procedure described in 3a-4 to give the target compound as a colorless transparent oil in 87% yield. 1 H NMR (400 MHz, CDC13) δ = 7.97 (d, J = 1.6 Hz, 1H), 7.82 (d, J = 1.6 Hz, 1H), 7.47-7.41 (m, 6H), 7.33-7.28 (m, 6H), 7.26-7.21 (m, 3H), 5.44-5.34 (m, 1H), 4.63-4.54 (m, 1H), 4.54-4.45 (m, 1H), 3.42-3.25 (m, 2H), 3.06-2.89 (m, 2H), 2.35 (t, J = 7.6 Hz, 2H), 1.62 (br dd, J = 7.2, 14.8 Hz, 2H), 1.50-1.41 (m, 2H), 1.40-1.20 (m, 52H), 0.92-0.86 (m, 6H). HRMS (ESI): calcd for C 62 H 88 BrClO5[M+Na] + 1049.5396, found 1049.5382.

[0561] (S)-3-hydroxy-2-(stearoyloxy)propyl 3-bromo-5-chloro-4-pentadecylbenzoate 4a-17

[0562]

[0563] Following the procedure described in 4a-1, 4a-17 was treated to give the target compound as a white solid in 86% yield. 1 H NMR (400 MHz, CDC13) δ = 8.08 (d, J = 1.6 Hz, 1H), 7.94 (d, J = 1.6 Hz, 1H), 5.24 (quin, J = 5.2 Hz, 1H), 4.62-4.44 (m, 2H), 3.83 (t, J = 5.2 Hz, 2H), 3.02-2.92 (m, 2H), 2.37 (t, J = 7.6 Hz, 2H), 2.01 (t, J = 6.4 Hz, 1H), 1.68-1.60 (m, 2H), 1.58-1.52 (m, 2H), 1.48-1.40 (m, 2H), 1.36-1.22 (m, 50H), 0.92-0.86 (m, 6H). HRMS (ESI): calcd for C 43 H 74 BrClO5[M+Na] + 807.4300, found 807.4315.

[0564] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 3-bromo-5-chloro-4-pentadecyl-benzoate 5a-17

[0565]

[0566] Following the procedure described in 5a-1, 4a-17 was treated to give the target compound as a colorless transparent oil in 16% yield. 1HNMR (400 MHz, CDC13) δ = 8.07 (d, J = 1.6 Hz, 1H), 7.93 (d, J = 1.6 Hz, 1H), 5.53-5.42 (m, 1H), 5.32 (br dd, J = 3.6, 6.0 Hz, 1H), 4.64-4.51 (m, 1H), 4.46-4.37 (m, 1H), 4.35-4.25 (m, 1H), 4.21-4.06 (m, 2H), 4.03-3.88 (m, 2H), 3.01-2.92 (m, 2H), 2.34 (t, J = 7.6 Hz, 2H), 1.66-1.60 (m, 2H), 1.55 (br d, J = 3.2 Hz, 2H), 1.48-1.45 (m, 18H), 1.42 (d, J = 6.8 Hz, 9H), 1.33-1.21 (m, 52H), 0.91-0.87 (m, 6H). HRMS (ESI): calcd for C 59 H 104 BrClNO 11 P[M+Na] + 1170.6111, found 1170.6116.

[0567] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-bromo-5-chloro-4-pentadecyl-benzoate6a-17

[0568]

[0569] Following the procedure described in 6a-1, 5a-17 was treated to give the target compound as a colorless transparent oil in 39% yield. 1HNMR (400 MHz, CDC13) δ = 8.07 (s, 1H), 7.93 (s, 1H), 5.60-5.46 (m, 1H), 5.42-5.30 (m, 1H), 4.59 (td, J = 3.6, 12.0 Hz, 1H), 4.48-4.33 (m, 3H), 4.29-4.16 (m, 3H), 3.04-2.91 (m, 2H), 2.35 (dt, J = 3.6, 7.6 Hz, 2H), 1.65-1.57 (m, 6H), 1.53-1.47 (m, 18H), 1.45 (s, 9H), 1.30-1.22 (m, 48H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 104 BrClNO 12 P[M+Na] + 1186.6060, found 1186.6078.

[0570] O-(((R)-3-((3-bromo-5-chloro-4-pentadecylbenzoyl)oxy)-2-(stearoyloxy)propoxy)(hydroxy)phosphoryl)-D-serine A17

[0571]

[0572] According to the method described in Al, 6a-17 was treated to obtain the target compound, white solid, yield 44%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.01 (s, 1H), 7.87 (br s, 1H), 7.78 (br s, 2H), 5.44 (br s, 1H), 4.60 (br d, J = 8.0 Hz, 3H), 4.47 (br s, 2H), 4.19 (br s, 2H), 3.06-2.88 (m, 2H), 2.40 (br s, 2H), 1.58 (br d, J = 5.2 Hz, 4H), 1.45 (br s, 2H), 1.27 (br s, 50H), 0.89 (br t, J = 6.4 Hz, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 176.43, 168.89, 165.30, 146.80, 135.35, 132.45, 129.73, 128.18, 125.57, 70.42, 70.36, 64.91, 63.68, 63.45, 54.24, 34.57, 34.31, 31.93, 29.71, 29.67, 29.58, 29.37, 29.27, 29.11, 27.73, 24.84, 22.68, 14.05. HRMS (ESI): calcd for C 46 H 80 BrClNO 10 P[M-H] - 950.4319, 952.4299, found 952.4403.

[0573] Synthesis of compound A18 of Example 18

[0574] 4-(tetradecylamino)benzoic acid 1a-12

[0575]

[0576] Following the procedure described in 1a-3, 4-aminobenzoic acid and 1- bromotetradecane were treated to give the target compound as a light yellow solid in 19% yield. 1 H NMR (400 MHz, CDC13) δ 7.91 (dd, J = 8.8, 2.0 Hz, 2H), 6.55 (dd, J = 8.6, 1.9 Hz, 2H), 3.17 (t, J = 6.9 Hz, 2H), 1.64 (p, J = 7.6 Hz, 2H), 1.28 (d, J = 15.8 Hz, 22H), 0.88 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 43 H 55 NO4[M+H] + 334.2741, found 334.2748.

[0577] (S)-2-hydroxy-3-(trityloxy)propyl 4-(tetradecylamino)benzoate 2a-10

[0578]

[0579] Following the procedure described in 2a-1, 1a-12 and 1a were treated to give the target compound as a white solid in 82% yield.1 HNMR (400 MHz, cdcl3) δ 7.78 (d, J = 8.3 Hz, 2H), 7.45 (d, J = 7.7 Hz, 6H), 7.26 (dp, J = 19.4, 6.8 Hz, 9H), 6.51 (d, J = 8.3 Hz, 2H), 4.44 - 4.35 (m, 2H), 4.10 (d, J = 8.7 Hz, 2H), 3.30 (d, J = 5.2 Hz, 2H), 3.16 (q, J = 6.1 Hz, 2H), 2.65 - 2.54 (m, 1H), 1.64 (q, J = 7.2 Hz, 2H), 1.45 - 1.21 (m, 22H), 0.88 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 43 H 55 NO4[M + Na] + 673.3863, found 673.3845.

[0580] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 4-(tetradecylamino)benzoate 3a-18

[0581]

[0582] Stearic acid and 2a-10 were treated according to the procedure described in 3a-4 to give the target compound as a white solid in 87% yield. 1 HNMR (400 MHz, cdcl3) δ 7.73 (d, J = 8.4 Hz, 2H), 7.44 (d, J = 7.6 Hz, 6H), 7.25 (dt, J = 24.0, 7.4 Hz, 9H), 6.49 (d, J = 8.6 Hz, 2H), 5.39 (p, J = 5.3 Hz, 1H), 4.51 - 4.42 (m, 2H), 4.09 (t, J = 5.0 Hz, 1H), 3.31 (qd, J = 9.9, 4.9 Hz, 2H), 3.15 (q, J = 6.5 Hz, 2H), 2.33 (t, J = 7.6 Hz, 2H), 1.63 (p, J = 7.2 Hz, 4H), 1.23 (d, J = 17.4 Hz, 50H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 61 H 89 NO5[M + Na] + 938.6633, found 938.6625.

[0583] (S)-3-hydroxy-2-(stearoyloxy)propyl 4-(tetradecylamino)benzoate 4a-18

[0584]

[0585] Following the procedure described in 4a-1, 3a-18 was treated to give the target compound as a white solid in 46% yield. 1 H NMR (400 MHz, cdcl3) δ 7.84 (d, J = 8.1 Hz, 2H), 6.53 (d, J = 8.4 Hz, 2H), 4.46 (d, J = 5.1 Hz, 2H), 3.78 (t, J = 5.4 Hz, 2H), 3.16 (t, J = 7.1 Hz, 2H), 2.36 (t, J = 7.8 Hz, 3H), 1.63 (t, J = 7.3 Hz, 4H), 1.25 (d, J = 5.9 Hz, 50H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 42 H 75 NO5[M+Na] + 696.5537, found 696.5534.

[0586] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 4-(tetradecylamino)benzoate 5a-18

[0587]

[0588] Following the procedure described in 5a-1, 4a-18 was treated to give the target compound as a colorless transparent oil in 49% yield. 1HNMR (400 MHz, cdcl3) δ 7.82 (d, J = 8.5 Hz, 2H), 6.52 (d, J = 8.3 Hz, 2H), 5.53 (dd, J = 22.9, 8.3 Hz, 1H), 5.32 (t, J = 6.0 Hz, 1H), 4.46 (dt, J = 12.6, 4.5 Hz, 1H), 4.39 - 4.32 (m, 1H), 4.33 - 4.21 (m, 1H), 4.21 - 4.03 (m, 2H), 3.95 (dq, J = 22.0, 5.9 Hz, 2H), 3.15 (q, J = 6.6 Hz, 2H), 2.32 (t, J = 7.5 Hz, 2H), 1.61 (dd, J = 14.2, 7.3 Hz, 4H), 1.49 - 1.17 (m, 77H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 58 H 105 N2O 11 P[M+Na] + 1059.7348, found 1059.7346.

[0589] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 4-(tetradecylamino)benzoate 6a-18

[0590]

[0591] Following the procedure described for 6a-1, 5a-18 was treated to give the target compound as a colorless transparent oil in 43% yield. 1 HNMR (400 MHz, cdcl3) δ 7.82 (d, J = 8.6 Hz, 2H), 6.52 (d, J = 8.2 Hz, 2H), 5.55 (t, J = 9.1 Hz, 1H), 5.36 (q, J = 4.7 Hz, 1H), 4.48 (dd, J = 11.9, 4.6 Hz, 1H), 4.36 (dt, J = 11.5, 5.4 Hz, 3H), 4.19 (ddt, J = 24.3, 12.2, 6.1 Hz, 4H), 3.14 (d, J = 7.7 Hz, 2H), 2.33 (t, J = 7.5 Hz, 2H), 1.62 (q, J = 7.4 Hz, 4H), 1.54 - 1.17 (m, 77H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C58 H 105 N2O 12 P[M+Na] + 1075.7297, found 1075.7299.

[0592] O-(hydroxy((R)-2-(stearoyloxy)-3-((4-(tetradecylamino)benzoyl)oxy)propoxy)phosphoryl)-D-serine A18

[0593]

[0594] The compound 6a-18 was treated according to the method described in Al to give the target compound as a white solid in 57% yield. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.16 (br d, J = 8.4 Hz, 2H), 7.79 - 7.59 (m, 2H), 7.56 (br d, J = 8.4 Hz, 2H), 5.53 - 5.33 (m, 1H), 4.79 - 4.36 (m, 5H), 4.35 - 4.13 (m, 2H), 3.55 - 3.41 (m, 2H), 2.41 (br t, J = 7.6 Hz, 2H), 1.81 - 1.68 (m, 2H), 1.59 (br d, J = 6.8 Hz, 2H), 1.38 (br d, J = 6.8 Hz, 2H), 1.26 (br s, 48H), 0.88 (t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.04, 168.67, 165.30, 138.66, 132.04, 130.99, 122.99, 70.13, 64.71, 63.41, 63.16, 54.09, 53.97, 31.91, 29.70, 29.63, 29.51, 29.35, 29.21, 28.94, 28.81, 26.04, 25.66, 22.66, 14.04. HRMS (ESI): calcd for C 45 H 81 N2O 10 P[M-H] - 839.5556, found 839.5546.

[0595] Synthesis of compound A19 of Example 19

[0596] (S)-2-hydroxy-3-(trityloxy)propyl 4-pentadecyl-3-(trifluoromethyl)benzoate 2a-11

[0597]

[0598] The target compound was obtained in 46% yield as colorless transparent oil by treating 4-pentadecyl-3-(trifluoromethyl)benzoic acid and 1a according to the method described in 2a-1. 1 H NMR (400 MHz, CDC13) δ = 8.20 (s, 1H), 8.05-8.00 (m, 1H), 7.47-7.43 (m, 6H), 7.40 (d, J = 8.0 Hz, 1H), 7.30 (s, 6H), 7.25 (d, J = 7.2 Hz, 3H), 4.46 (d, J = 4.8 Hz, 2H), 4.18-4.13 (m, 1H), 3.33 (t, J = 4.8 Hz, 2H), 2.87-2.77 (m, 2H), 2.52-2.38 (m, 1H), 1.68-1.59 (m, 2H), 1.45-1.38 (m, 2H), 1.31-1.25 (m, 22H), 0.91-0.87 (m, 3H). HRMS (ESI): calcd for C 45 H 55 F3O4[M+Na] + 739.3945, found 739.3962.

[0599] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 4-pentadecyl-3-(trifluoromethyl)benzoate 3a-19

[0600]

[0601] The target compound was obtained in 100% yield as colorless transparent oil by treating stearic acid and 2a-11 according to the method described in 3a-4. 1H NMR (400 MHz, CDC13) δ = 8.17 (s, 1H), 7.98 (d, J = 8.0 Hz, 1H), 7.48-7.41 (m, 6H), 7.39 (d, J = 8.0 Hz, 1H), 7.32-7.27 (m, 6H), 7.25-7.19 (m, 3H), 5.48-5.36 (m, 1H), 4.63-4.45 (m, 2H), 3.33 (tt, J = 5.2, 10.4 Hz, 2H), 2.87-2.77 (m, 2H), 2.35 (t, J = 7.6 Hz, 2H), 1.69-1.58 (m, 4H), 1.46-1.38 (m, 2H), 1.32-1.21 (m, 50H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 63 H 89 F3O5[M+Na] + 1005.6554, found 1005.6548.

[0602] (S)-3-hydroxy-2-(stearoyloxy)propyl 4-pentadecyl-3-(trifluoromethyl)benzoate 4a-19

[0603]

[0604] Following the procedure described in 4a-1, 3a-19 was treated to give the target compound as a white solid in 59% yield. 1 H NMR (400 MHz, CDC13) δ = 8.17 (s, 1H), 7.98 (d, J = 8.0 Hz, 1H), 7.48-7.41 (m, 6H), 7.39 (d, J = 8.0 Hz, 1H), 7.32-7.27 (m, 6H), 7.25-7.19 (m, 3H), 5.48-5.36 (m, 1H), 4.63-4.45 (m, 2H), 3.33 (tt, J = 5.2, 10.4 Hz, 2H), 2.87-2.77 (m, 2H), 2.35 (t, J = 7.6 Hz, 2H), 1.69-1.58 (m, 4H), 1.46-1.38 (m, 2H), 1.32-1.21 (m, 50H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 44 H 75 F3O5[M+Na] + 763.5459, found 763.5466.

[0605] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 4-pentadecyl-3- (trifluoromethyl)benzoate 5a-19

[0606]

[0607] Following the procedure described in 5a-1, 4a-19 was treated to give the target compound as a colorless transparent oil in 35% yield. 1 HNMR (400 MHz, CDC13) δ = 8.26 (s, 1H), 8.09 (br d, J = 8.0 Hz, 1H), 7.42 (d, J = 8.0 Hz, 1H), 5.48 (br d, J = 8.0 Hz, 1H), 5.39-5.31 (m, 1H), 4.71-4.28 (m, 4H), 4.12-3.87 (m, 3H), 2.85-2.77 (m, 2H), 2.34 (br t, J = 7.5 Hz, 2H), 1.66-1.58 (m, 4H), 1.49-1.44 (m, 18H), 1.41 (d, J = 7.2 Hz, 9H), 1.31-1.20 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 60 H 105 F3NO 11 P[M+Na] + 1126.7270, found 1126.7264.

[0608] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 4-pentadecyl-3- (trifluoromethyl)benzoate 5a-19

[0609]

[0610] Following the procedure described in 6a-1, 5a-19 was treated to give the target compound as a colorless transparent oil in 70% yield. 1HNMR (400 MHz, CDC13) δ = 8.27 (s, 1H), 8.11 (br d, J = 8.4 Hz, 1H), 7.44 (d, J = 8.4 Hz, 1H), 5.52 (br t, J = 7.6 Hz, 1H), 5.45-5.37 (m, 1H), 4.62 (td, J = 4.0, 12.0 Hz, 1H), 4.51-4.34 (m, 3H), 4.32-4.17 (m, 3H), 2.91-2.76 (m, 2H), 2.36 (dt, J = 3.6, 7.6 Hz, 2H), 1.63 (br d, J = 7.6 Hz, 4H), 1.55-1.41 (m, 27H), 1.37-1.18 (m, 52H), 0.90 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 60 H 105 F3NO 12 P[M+Na] + 1142.7219, found 1142.7225.

[0611] O-(hydroxy((R)-3-((4-pentadecyl-3-(trifluoromethyl)benzoyl)oxy)-2-(stearoyloxy)propoxy)phosphoryl)-D-serine A19

[0612]

[0613] According to the method described in Al, 6a-19 was treated to obtain the target compound, light yellow solid, yield 75%. 1 H NMR (400 MHz, CDC13) δ = 8.16 (br s, 1H), 8.00 (br s, 1H), 7.33 (br s, 1H), 5.38 (br d, J = 5.1 Hz, 1H), 4.80-3.74 (m, 7H), 2.75 (br d, J = 1.1 Hz, 2H), 2.26 (br s, 2H), 1.62-1.46 (m, 4H), 1.34-1.13 (m, 52H), 0.88 (br t, J = 6.4 Hz, 6H). 13C NMR (101 MHz, CDC13) δ = 173.68, 165.22, 147.17, 132.65, 131.03, 128.65, 128.36, 128.06, 127.45, 127.25, 127.22, 125.33, 122.61, 70.39, 70.36, 64.79, 64.25, 63.87, 63.79, 55.55, 32.82, 31.93, 31.54, 30.01, 29.77, 29.70, 29.60, 29.39, 29.15, 24.94, 22.68, 14.05. HRMS (ESI): calcd for C 47 H 81 F3NO 10 P[M-H] - 906.5477, found 906.5464.

[0614] Synthesis of compound A20 of Example 20

[0615] methyl 3-chloro-5-fluoro-4-(tetradecylamino)benzoate 1a-13

[0616]

[0617] Methyl 3-chloro-4,5-difluorobenzoate (4.80 g, 23.2 mmol, 1.00 eq.) and tetradecan-1-amine (5.45 g, 25.6 mmol, 1.10 eq.) were dissolved in dimethyl sulfoxide (50.0 mL), and N,N-diisopropylethylamine (9.01 g, 69.7 mmol, 12.1 mL, 3.00 eq.) was added. The mixed solution was stirred at 110 °C for 16 h under nitrogen. The reaction mixture was diluted with water (100 mL) and extracted with EA (100 mL x 3), the organic layers were combined, washed with brine (100 mL x 3), dried over sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 100 / 0 ~ 15 / 1) to obtain the target compound as a white solid in 81% yield. 1 H NMR (400 MHz, CDC13) δ = 173.68, 165.22, 147.17, 132.65, 131.03, 128.65, 128.36, 128.06, 127.45, 127.25, 127.22, 125.33, 122.61, 70.39, 70.36, 64.79, 64.25, 63.87, 63.79, 55.55, 32.82, 31.93, 31.54, 30.01, 29.77, 29.70, 29.60, 29.39, 29.15, 24.94, 22.68, 14.05. HRMS (ESI): calcd for C 22H 35 ClFNO2[M+H] + 400.2413, found 400.2405.

[0618] 3-chloro-5-fluoro-4-(tetradecylamino)benzoic acid 1a-14

[0619]

[0620] Methyl 3-chloro-5-fluoro-4-(tetradecylamino)benzoate (7.49 g, 18.70 mmol, 1.00 eq.) was dissolved in a solution of tetrahydrofuran (80.00 mL), water (40.00 mL) and methanol (40.00 mL), lithium hydroxide hydrate (3.14 g, 74.90 mmol, 4.00 eq.) was added, stirred at 20 °C for 14 h, diluted with water (100 mL) and hydrochloric acid (1.00 M, 44.00 mL) was added to adjust pH = 3. Then the mixture was extracted with EA (100 mL x 3), the combined organic layer was washed with brine (300 mL x 3), dried over sodium sulfate, concentrated under reduced pressure to give the target compound, white solid, yield 97%. 1 H NMR (400 MHz, CDC13) δ = 7.84 (s, 1H), 7.63 (dd, J = 1.6, 13.6 Hz, 1H), 3.52 (dt, J = 3.2, 7.2 Hz, 2H), 1.63 (quin, J = 7.2 Hz, 2H), 1.42 - 1.26 (m, 22H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 21 H 33 ClFNO2[M+H] + 386.2257, found 386.2268.

[0621] (S)-2-hydroxy-3-(trityloxy)propyl 3-chloro-5-fluoro-4-(tetradecylamino)benzoate 2a-12

[0622]

[0623] According to the method described in 2a-1, 1a-14 and 1a were treated to give the target compound, colorless transparent oil, yield 53%. 1HNMR (400 MHz, CDC13) δ = 7.68 (dd, J = 1.2, 1.6 Hz, 1H), 7.50 (d, J = 2.0 Hz, 1H), 7.47-7.43 (m, 7H), 7.27 (s, 11H), 4.46 (br s, 1H), 4.43-4.34 (m, 2H), 4.14-4.06 (m, 1H), 3.51 (br d, J = 1.6 Hz, 2H), 3.35-3.27 (m, 2H), 2.47 (d, J = 5.6 Hz, 1H), 1.68-1.58 (m, 2H), 1.43-1.24 (m, 24H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 43 H 53 ClFNO4[M+Na] + 724.3539, found 724.3531.

[0624] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 3-chloro-5-fluoro-4-(tetradecylamino)benzoate3a-20

[0625]

[0626] According to the method described in 3a-4, stearic acid and 2a-12 were treated to obtain the target compound, colorless transparent oil, yield 97%. 1 H NMR (400 MHz, CDC13) δ = 7.64 (d, J = 0.8 Hz, 1H), 7.47-7.41 (m, 7H), 7.27 (s, 9H), 5.43-5.31 (m, 1H), 4.58-4.41 (m, 3H), 3.55-3.44 (m, 2H), 3.39-3.25 (m, 2H), 2.35 (t, J = 7.2 Hz, 2H), 1.68-1.59 (m, 4H), 1.50-1.14 (m, 48H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 61 H 87 ClFNO5[M+Na] + 990.6149, found 990.6155.

[0627] (S)-3-hydroxy-2-(stearoyloxy)propyl 3-chloro-5-fluoro-4-(tetradecylamino)benzoate 4a-20

[0628]

[0629] Following the procedure described in 4a-1, 3a-20 was treated to give the target compound as a white solid in 65% yield. 1 H NMR (400 MHz, CDC13) δ = 7.74 (s, 1H), 7.55 (dd, J = 1.6, 13.6 Hz, 1H), 5.21 (t, J = 5.2 Hz, 1H), 4.54-4.40 (m, 3H), 3.80 (t, J = 5.6 Hz, 2H), 3.57-3.44 (m, 2H), 2.37 (t, J = 7.6 Hz, 2H), 2.15 (t, J = 6.4 Hz, 1H), 1.70-1.60 (m, 4H), 1.42-1.21 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 42 H 73 ClFNO5[M+Na] + 748.5054, found 748.5045.

[0630] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(stearoyloxy)propyl 3-chloro-5-fluoro-4-(tetradecyl-amino)benzoate 5a-20

[0631]

[0632] Following the procedure described in 5a-1, 4a-20 was treated to give the target compound as a colorless transparent oil in 28% yield. 1HNMR (400 MHz, CDC13) δ = 7.73 (s, 1H), 7.54 (dd, J = 1.6, 13.6 Hz, 1H), 5.49 (br d, J = 7.6 Hz, 1H), 5.36-5.23 (m, 1H), 4.56-4.27 (m, 4H), 4.12-3.86 (m, 4H), 3.49 (dq, J = 3.2, 6.8 Hz, 2H), 2.33 (t, J = 7.6 Hz, 2H), 1.66-1.59 (m, 4H), 1.52-1.39 (m, 27H), 1.35-1.18 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 58 H 103 ClFN2O 11 P[M+Na] + 1111.6864, found 1111.6849.

[0633] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-chloro-5-fluoro-4-(tetradecyl-amino)benzoate 6a-20

[0634]

[0635] According to the method described in 6a-1, 5a-20 was treated to obtain the target compound, colorless transparent oil, yield 53%. 1 HNMR (400 MHz, CDC13) δ = 7.73 (s, 1H), 7.54 (dd, J = 1.6, 13.6 Hz, 1H), 5.49 (br d, J = 7.6 Hz, 1H), 5.36-5.23 (m, 1H), 4.56-4.27 (m, 4H), 4.12-3.86 (m, 4H), 3.49 (dq, J = 3.2, 6.8 Hz, 2H), 2.33 (t, J = 7.6 Hz, 2H), 1.66-1.59 (m, 4H), 1.52-1.39 (m, 27H), 1.35-1.18 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 58 H 103 ClFN2O 12 P[M+Na] +1127.6813, found 1127.6822.

[0636] O-(((R)-3-((3-chloro-5-fluoro-4-(tetradecylamino)benzoyl)oxy)-2-(stearoyloxy)propoxy)(hydroxy)phosphoryl)-D-serine A20

[0637]

[0638] According to the method described in A1, 6a-20 was treated to give the target compound, a light yellow solid, yield 62%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.92 (s, 1H), 7.75 (br d, J = 10.4 Hz, 4H), 5.38 (br s, 1H), 4.68 - 4.34 (m, 5H), 4.17 (br s, 2H), 3.54 (br t, J = 7.6 Hz, 2H), 2.39 (br t, J = 7.2 Hz, 2H), 1.81 - 1.70 (m, 2H), 1.66 - 1.54 (m, 2H), 1.40 (br s, 2H), 1.26 (s, 48H), 0.88 (t, J = 6.8 Hz, 6H). 31 P NMR (162 MHz, CDC13 and 20 μL TFA) δ = -1.93. 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 175.95, 168.77, 164.06, 161.29, 160.86, 160.45, 160.03, 155.14, 152.63, 129.82, 127.55, 127.00, 126.54, 118.79, 117.30, 117.07, 115.95, 113.12, 110.29, 70.13, 64.64, 63.49, 54.14, 50.28, 34.17, 31.91, 29.71, 29.62, 29.45, 29.34, 29.13, 29.00, 26.28, 24.74, 22.67, 14.04. HRMS (ESI): calcd for C 45 H 79 ClFN2O 10 P[M-H] - 891.5072, found 891.5074.

[0639] Synthesis of compound A21 of Example 21

[0640] 4-pentadecylbenzoic acid 1a-15

[0641]

[0642] 4-methylbenzoic acid and 1-bromotetradecane were treated according to the procedure described in 1a-9 to give the target compound as a white solid in 14% yield. 1 H NMR (400 MHz, DMSO-d6) δ = 13.01 - 12.52 (m, 1H), 7.85 (d, J = 8.0 Hz, 2H), 7.30 (d, J = 8.0 Hz, 2H), 2.63 (t, J = 7.6 Hz, 2H), 1.65 - 1.50 (m, 2H), 1.35 - 1.15 (m, 24H), 0.85 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 22 H 36 O2[M+H] + 333.2788, found 333.2769.

[0643] 3-nitro-4-pentadecylbenzoic acid 1a-16

[0644]

[0645] 4-pentadecylbenzoic acid (15.00 g, 45.10 mmol, 1.00 eq.) was added to trifluoroacetic acid (350 mL) at 25 °C, fuming nitric acid (11.40 g, 180 mmol, 8.12 mL, 4.00 eq.) was added, stirred at 40 °C for 1 h, the resulting solid was filtered, washed with water, and triturated with toluene and dichloromethane to remove water and trifluoroacetic acid to give the target compound as a white solid in 53% yield. 1 H NMR (400 MHz, DMSO-d6) δ = 8.35 (d, J = 1.2 Hz, 1H), 8.13 (dd, J = 1.6, 8.0 Hz, 1H), 7.65 (d, J = 8.0 Hz, 1H), 2.91 - 2.81 (m, 2H), 1.64 - 1.49 (m, 2H), 1.34 - 1.14 (m, 24H), 0.85 (br t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 22 H 35 NO4[M+H] + 378.2639, found 378.2644.

[0646] (S)-2-hydroxy-3-(trityloxy)propyl 3-nitro-4-pentadecylbenzoate 2a-13

[0647]

[0648] Following the procedure described in 2a-1, 1a-16 and 1a were treated to give the target compound as a colorless transparent oil in 24% yield. 1 HNMR (400 MHz, CDC13) δ = 8.39 (d, J = 1.2 Hz, 1H), 8.08 (dd, J = 1.6, 8.0 Hz, 1H), 7.48-7.41 (m, 7H), 7.34-7.29 (m, 6H), 7.26 (br d, J = 7.2 Hz, 3H), 4.48 (d, J = 5.2 Hz, 2H), 4.15 (d, J = 7.2 Hz, 1H), 3.43-3.26 (m, 2H), 3.00-2.88 (m, 2H), 2.47 (br d, J = 5.6 Hz, 1H), 1.72-1.63 (m, 2H), 1.36-1.27 (m, 24H), 0.90 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 44 H 55 NO6[M+Na] + 716.3922, found 716.3941.

[0649] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 3-nitro-4-pentadecylbenzoate 3a-21

[0650]

[0651] Following the procedure described in 3a-4, stearic acid and 2a-13 were treated to give the target compound as a white solid in 86% yield. 1HNMR (400 MHz, CDC13) δ = 8.35 (d, J = 1.6 Hz, 1H), 8.04 (dd, J = 1.6, 8.0 Hz, 1H), 7.48-7.40 (m, 7H), 7.35-7.29 (m, 6H), 7.28-7.22 (m, 3H), 5.48-5.38 (m, 1H), 4.67-4.52 (m, 2H), 3.35 (tt, J = 5.2, 10.0 Hz, 2H), 3.00-2.87 (m, 2H), 2.45-2.32 (m, 2H), 1.71-1.60 (m, 5H), 1.46-1.24 (m, 51H), 0.92-0.87 (m, 6H). HRMS (ESI): calcd for C 62 H 89 NO7[M+Na] + 982.6531, found 982.6535.

[0652] (S)-3-hydroxy-2-(stearoyloxy)propyl 3-nitro-4-pentadecylbenzoate 4a-21

[0653]

[0654] Following the procedure described in 4a-1, 3a-21 was treated to give the target compound as a white solid in 63% yield. 1 H NMR (400 MHz, CDC13) δ = 8.49 (br d, J = 1.6 Hz, 1H), 8.14 (br dd, J = 2.0, 8.0 Hz, 1H), 7.46 (br d, J = 8.0 Hz, 1H), 5.43-5.16 (m, 1H), 4.76-4.42 (m, 2H), 3.85 (br d, J = 4.8 Hz, 2H), 3.11-2.81 (m, 2H), 2.56-2.30 (m, 2H), 2.22-2.00 (m, 1H), 1.69-1.59 (m, 4H), 1.27 (br s, 52H), 0.99-0.86 (m, 6H). HRMS (ESI): calcd for C 43 H 75 NO7[M+Na] + 740.5436, found 740.5452.

[0655] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-nitro-4-pentadecylbenzoate 5a-21

[0656]

[0657] Following the procedure described in 5a-1, 4a-21 was treated to give the target compound as a colorless transparent oil in 17% yield. 1 HNMR (400 MHz, CDC13) δ = 8.48 (d, J = 1.6 Hz, 1H), 8.13 (dd, J = 1.2, 8.0 Hz, 1H), 7.44 (d, J = 8.0 Hz, 1H), 5.60-5.42 (m, 1H), 5.41-5.27 (m, 1H), 4.68-4.57 (m, 1H), 4.52-4.39 (m, 1H), 4.39-4.25 (m, 1H), 4.17-3.97 (m, 4H), 3.01-2.87 (m, 2H), 2.34 (t, J = 7.6 Hz, 2H), 1.70-1.60 (m, 4H), 1.46 (d, J = 8.8 Hz, 18H), 1.41 (d, J = 7.2 Hz, 9H), 1.33-1.21 (m, 52H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 105 N2O 13 P[M+Na] + 1103.7246, found 1103.7233.

[0658] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 3-nitro-4-pentadecylbenzoate 5a-21

[0659]

[0660] Following the procedure described in 5a-1, 4a-21 was treated to give the target compound as a colorless transparent oil in 17% yield. 1H NMR (400 MHz, CDC13) δ = 8.49 (s, 1H), 8.18-8.11 (m, 1H), 7.46 (d, J = 8.0 Hz, 1H), 5.63-5.32 (m, 2H), 4.64 (td, J = 4.0, 12.0 Hz, 1H), 4.54-4.43 (m, 1H), 4.42-4.38 (m, 1H), 4.32-4.17 (m, 3H), 3.00-2.89 (m, 2H), 2.37 (dt, J = 3.6, 7.6 Hz, 2H), 1.64 (br dd, J = 7.2, 14.8 Hz, 5H), 1.55-1.49 (m, 18H), 1.46 (d, J = 1.6 Hz, 9H), 1.36-1.23 (m, 52H), 0.93-0.86 (m, 6H). HRMS (ESI): calcd for C 59 H 105 N2O 14 P[M+Na] + 1119.7196, found 1119.7181.

[0661] O-(hydroxy((R)-3-((3-nitro-4-pentadecylbenzoyl)oxy)-2-(stearoyloxy)propoxy)phosphoryl)-D-serine A21

[0662]

[0663] The procedure described in Al was followed for the treatment of 6a-21 to give the target compound as a white solid in 71% yield. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.51-8.42 (m, 1H), 8.12 (br d, J = 8.0 Hz, 1H), 7.78 (br s, 2H), 7.63 (br s, 1H), 7.49 (d, J = 8.0 Hz, 1H), 5.48 (br s, 1H), 4.75-4.58 (m, 3H), 4.56-4.46 (m, 2H), 4.24 (br d, J = 6.0 Hz, 2H), 3.05-2.83 (m, 2H), 2.42 (br t, J = 7.2 Hz, 2H), 1.72-1.54 (m, 4H), 1.40 (br d, J = 7.6 Hz, 2H), 1.37-1.15 (m, 52H), 0.90 (t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.39, 168.92, 165.39, 149.20, 144.09, 133.39, 132.54, 127.72, 126.00, 70.33, 64.92, 63.65, 63.40, 54.30, 34.27, 33.17, 31.92, 29.71, 29.66, 29.58, 29.38, 29.36, 29.08, 28.94, 24.77, 22.67, 14.05. HRMS (ESI): calcd for C 46 H 81 N2O 12 P[M+H] + 885.5600, found 885.5587.

[0664] Synthesis of compound A22 of example 22

[0665] (S)-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)-3-(trityloxy)propyl 4-(tetradecyloxy)benzoate 3a-22

[0666]

[0667] Linoleic acid and 2a-4 were treated according to the procedure described for 3a-4 to give the target compound as a clear oil in 79% yield. 1 H NMR (400 MHz, CDC13) δ 7.88 - 7.81 (m, 2H), 7.46 - 7.40 (m, 6H), 7.28 (dd, J = 8.3, 6.4 Hz, 6H), 7.25 - 7.20 (m, 3H), 6.89 - 6.82 (m, 2H), 5.44 - 5.27 (m, 5H), 4.57 - 4.43 (m, 2H), 4.00 (t, J = 6.5 Hz, 2H), 3.32 (qd, J = 9.9, 5.1 Hz, 2H), 2.76 (t, J = 6.2 Hz, 2H), 2.38 - 2.27 (m, 2H), 2.03 (dq, J = 12.9, 6.7 Hz, 4H), 1.80 (p, J = 6.7 Hz, 2H), 1.62 (d, J = 7.2 Hz, 2H), 1.45 (q, J = 7.1 Hz, 2H), 1.39 - 1.19 (m, 36H), 0.91 - 0.85 (m, 6H). HRMS (ESI): calcd for C 61 H 84 O6[M+Na] +935.616, found 935.6154.

[0668] (S)-3-hydroxy-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propyl 4-(tetradecyloxy)benzoate 4a-22

[0669]

[0670] Following the procedure described in 4a-1, 3a-22 was treated to give the target compound as a white solid in 60% yield. 1 H NMR (400 MHz, cdcl3) δ 8.01 - 7.93 (m, 2H), 6.93 - 6.87 (m, 2H), 5.43 - 5.27 (m, 4H), 4.45 - 4.32 (m, 2H), 4.32 - 4.15 (m, 3H), 4.00 (t, J = 6.6 Hz, 2H), 2.77 (t, J = 6.4 Hz, 2H), 2.35 (t, J = 7.6 Hz, 2H), 2.04 (q, J = 6.9 Hz, 4H), 1.84 - 1.74 (m, 2H), 1.64 (d, J = 7.2 Hz, 2H), 1.44 (q, J = 6.8 Hz, 2H), 1.40 - 1.22 (m, 34H), 0.88 (td, J = 6.9, 3.3 Hz, 6H). HRMS (ESI): calcd for C 42 H 70 O6[M+H] + 671.5245, found 671.5233.

[0671] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propyl 4-(tetradecyloxy)benzoate5a-22

[0672]

[0673] Following the procedure described in 5a-1, 4a-22 was treated to give the target compound as a colorless transparent oil in 68% yield. 1HNMR (400 MHz, cdcl3) δ 7.97 (d, J = 8.7 Hz, 2H), 6.88 (dd, J = 8.8, 3.5 Hz, 2H), 5.57 (d, J = 8.1 Hz, 1H), 5.32 (tt, J = 11.1, 5.5 Hz, 4H), 4.80 (dp, J = 9.9, 5.1 Hz, 1H), 4.52 - 4.19 (m, 6H), 3.97 (t, J = 6.6 Hz, 2H), 2.74 (t, J = 6.4 Hz, 2H), 2.32 (q, J = 8.0 Hz, 2H), 2.02 (d, J = 7.0 Hz, 4H), 1.77 (p, J = 6.8 Hz, 2H), 1.60 (d, J = 6.9 Hz, 2H), 1.49 - 1.38 (m, 29H), 1.37 - 1.13 (m, 34H), 0.85 (td, J = 6.8, 3.4 Hz, 6H). HRMS (ESI): calcd for C 58 H 100 NO 12 P[M+Na] + 1056.6875, found 1056.6872.

[0674] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propyl 4-(tetradecyloxy)benzoate 6a-22

[0675]

[0676] Following the procedure described in 6a-1, 5a-22 was treated to give the target compound as a colorless transparent oil in 60% yield. 1HNMR (400 MHz, cdcl3) δ 8.00 - 7.94 (m, 2H), 6.88 (dd, J = 8.8, 3.5 Hz, 2H), 5.57 (d, J = 8.1 Hz, 1H), 5.31 (dtt, J = 17.8, 11.3, 5.7 Hz, 4H), 4.80 (dp, J = 9.9, 5.1 Hz, 1H), 4.52 - 4.16 (m, 6H), 3.97 (t, J = 6.6 Hz, 2H), 2.74 (t, J = 6.4 Hz, 2H), 2.32 (q, J = 8.0 Hz, 2H), 2.02 (d, J = 7.0 Hz, 4H), 1.77 (p, J = 6.8 Hz, 2H), 1.60 (d, J = 6.9 Hz, 2H), 1.49 - 1.38 (m, 29H), 1.37 - 1.18 (m, 34H), 0.85 (td, J = 6.8, 3.4 Hz, 6H). RMS (ESI): calcd for C 58 H 100 NO 13 P[M+Na] + 1072.6824, found 1072.6828.

[0677] O-(hydroxy((R)-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)-3-((4(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serine A22

[0678]

[0679] Following the procedure described for A11, 6a-22 was treated to give the target compound as a yellow solid in 34% yield. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.93 (br d, J = 8.8 Hz, 2H), 7.69 (br s, 2H), 6.93 (br d, J = 8.8 Hz, 2H), 5.62-5.21 (m, 2H), 5.07 (br d, J = 5.6 Hz, 1H), 4.84 (br s, 1H), 4.74-4.41 (m, 6H), 4.36 (br dd, J = 4.4, 11.2 Hz, 1H), 4.03 (br t, J = 6.5 Hz, 2H), 2.78 (br t, J = 6.0 Hz, 1H), 2.42 (br t, J = 7.6 Hz, 3H), 2.12-1.91 (m, 2H), 1.86-1.77 (m, 2H), 1.69-1.56 (m, 3H), 1.49-1.42 (m, 2H), 1.27 (br s, 36H), 0.89 (br t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 177.10, 168.79, 168.47, 164.26, 135.62, 134.24, 132.14, 130.30, 128.11, 123.22, 122.49, 119.91, 114.63, 79.82, 73.49, 68.65, 63.57, 63.46, 54.26, 37.10, 34.13, 31.92, 29.66, 29.59, 29.35, 28.97, 28.93, 27.17, 25.88, 24.63, 24.54, 22.68, 14.03. HRMS (ESI): calcd for C 45 H 76 NO 11 P[M-H] - 836.5083, found 836.5091.

[0680] Synthesis of compound A23 of Example 23

[0681] (S)-2-hydroxy-3-(trityloxy)propyl 2-(((dodecylthio)carbonothioyl)thio)propanoate 2a-14

[0682]

[0683] Following the procedure described in 2a-1, 1a and 2-(((dodecylthio)thiocarbonoyl)thio)propanoic acid were treated to give the target compound as yellow oil in 62% yield.1 H NMR(400MHz,cdcl3)δ7.42(d,J=7.3Hz,6H),7.35–7.21(m,9H),4.78(q,J=7.4 Hz,1H),4.31–4.20(m,2H),4.00(q,J=5.3Hz,1H),3.32(dd,J=8.9,6.3Hz,2H) ,3.22(dd,J=5.4,2.9Hz,2H),1.73–1.61(m,2H),1.55(d,J=7.4Hz,3H),1.38( dq,J=12.7,7.1Hz,2H),1.25(s,16H),0.88(t,J=6.7Hz,3H).RMS(ESI):calcd forC 38 H 50 O4S3[M+Na] + 689.2763,found 689.2760.

[0684] (4S)-8-methyl-7-oxo-1,1,1-triphenyl-10-thioxo-2,6-dioxa-9,11-dithiatricosan-4-yl(9Z,12Z)-octadeca-9,12-dienoate 3a-23

[0685]

[0686] Linoleic acid and 2a-14 were treated according to the method described in 3a-4 to obtain the target compound as a yellow oil in 85% yield. 1 HNMR(400MHz,cdcl3)δ7.42(d,J=7.4Hz,6H),7.34–7.27(m,7H),7.23(d,J=7.2Hz,2H),5.43–5.28(m,4H),5. 24(dt,J=9.0,4.5Hz,1H),4.76(qd,J=7.4,2.5Hz,1H),4.42(ddd,J=11.8,5.3,3.7Hz,1H),4.31(ddd,J=11.8, 8.1,6.5Hz,1H),3.37–3.19(m,4H),2.77(t,J=6.4Hz,2H),2.34(tt,J=7.3,2.6Hz,2H),2.10–2.00(m,4H),1.7 4–1.58(m,4H),1.51(dd,J=7.4,1.6Hz,3H),1.44–1.19(m,32H),0.88(td,J=6.9,2.9Hz,6H).RMS(ESI):calcd forC 56 H80 O5S3[M+Na] + 951.506, found 951.5055.

[0687] (2S)-1-((2-(((dodecylthio)carbonothioyl)thio)propanoyl)oxy)-3-hydroxypropan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 4a-23

[0688]

[0689] Following the procedure described in 4a-1, 3a-23 was treated to give the target compound as a yellow oil in 31% yield. 1 H NMR (400 MHz, cdcl3) δ 5.44 - 5.29 (m, 4H), 5.09 (dq, J = 7.9, 4.9 Hz, 1H), 4.86 - 4.76 (m, 1H), 4.45 - 4.17 (m, 2H), 3.74 (d, J = 5.0 Hz, 2H), 3.35 (td, J = 7.3, 3.2 Hz, 2H), 2.77 (t, J = 6.5 Hz, 2H), 2.35 (td, J = 7.6, 2.0 Hz, 2H), 2.05 (q, J = 6.8 Hz, 4H), 1.76 - 1.53 (m, 7H), 1.44 - 1.21 (m, 32H), 0.89 (td, J = 6.8, 4.0 Hz, 6H). RMS (ESI): calcd for C 37 H 66 O5S3[M+Na] + 709.3965, found 709.3960.

[0690] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-3-((2-(((dodecylthio)carbonothioyl)thio)propanoyl)oxy)propan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 5a-23

[0691]

[0692] Following the procedure described in 5a-1, 4a-23 was treated to give the target compound as a yellow oil in 42% yield.1 H NMR (400 MHz, cdcl3) δ 5.59 - 5.28 (m, 4H), 5.18 (s, 1H), 4.43 (dd, J = 11.8, 3.6 Hz, 1H), 4.30 (d, J = 8.9 Hz, 1H), 4.20 (ddd, J = 23.0, 12.0, 6.4 Hz, 1H), 4.06 (s, 1H), 4.00 - 3.78 (m, 2H), 3.40 - 3.27 (m, 2H), 2.77 (t, J = 6.5 Hz, 2H), 2.32 (t, J = 7.8 Hz, 2H), 2.05 (q, J = 7.0 Hz, 4H), 1.68 (p, J = 7.5 Hz, 2H), 1.62 - 1.56 (m, 5H), 1.51 - 1.23 (m, 59H), 0.89 (td, J = 6.9, 4.1 Hz, 6H). RMS (ESI): calcd for C 53 H 96 NO 11 PS3[M+Na] + 1072.5775, found 1072.5773.

[0693] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-3-((2-(((dodecylthio)carbonothioyl)thio)propanoyl)oxy)propan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 6a-23

[0694]

[0695] Following the procedure described in 6a-1, 5a-23 was treated to give the target compound as a yellow oil in 42% yield. 1H NMR (400 MHz, cdcl3) δ 5.62 - 5.30 (m, 4H), 5.23 (dd, J = 14.1, 5.1 Hz, 1H), 4.82 (dtd, J = 7.5, 4.8, 2.4 Hz, 1H), 4.48 - 4.29 (m, 2H), 4.16 (ddt, J = 47.2, 11.6, 5.6 Hz, 3H), 3.42 - 3.27 (m, 2H), 2.77 (t, J = 6.5 Hz, 2H), 2.33 (t, J = 7.6 Hz, 2H), 2.05 (q, J = 6.9 Hz, 4H), 1.68 (p, J = 7.4 Hz, 2H), 1.64 - 1.59 (m, 3H), 1.50 - 1.43 (m, 27H), 1.41 - 1.23 (m, 32H), 0.88 (td, J = 6.9, 4.0 Hz, 6H). RMS (ESI): calcd for C 53 H 96 NO 12 PS3[M+Na] + 1088.5724, found 1088.5723.

[0696] O-(((2R)-3-((2-(((dodecylthio)carbonothioyl)thio)propanoyl)oxy)-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serineA23

[0697]

[0698] Following the procedure described for A11, 6a-23 was treated to give the target compound as a yellow oil in 22% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 7.69 (br s, 3H), 5.68 - 5.01 (m, 4H), 4.80 (br dd, J = 4.8, 7.6 Hz, 1H), 4.59 (br d, J = 1.2 Hz, 2H), 4.53 - 4.37 (m, 3H), 4.31 - 4.09 (m, 4H), 3.46 - 3.22 (m, 2H), 2.51 - 2.27 (m, 4H), 2.00 (br dd, J = 6.4, 13.6 Hz, 2H), 1.72 - 1.67 (m, 2H), 1.59 (br d, J = 7.2 Hz, 3H), 1.31 - 1.25 (m, 36H), 0.89 (br t, J = 6.8 Hz, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 220.04, 173.69, 170.61, 167.03, 134.82, 132.35, 132.12, 129.50, 68.27, 62.87, 61.76, 52.31, 45.83, 35.61, 32.34, 30.09, 27.89, 27.80, 27.62, 27.51, 27.28, 27.10, 25.96, 22.87, 20.85, 14.34, 12.25. HRMS (ESI): calcd for C 40 H 72 NO 10 PS3[M-H] - 852.3983, found 852.3947.

[0699] Synthesis of compound A24 of Example 24

[0700] (S)-2-hydroxy-3-(trityloxy)propyl 3-nitro-4-(tetradecyloxy)benzoate 2a-15

[0701]

[0702] Following the procedure described in 2a-1, 1a and 1a-5 were treated, white solid, yield 63%. 1 H NMR (400 MHz, CDC13) δ 8.34 (t, J = 1.7 Hz, 1H), 8.11 (dd, J = 8.8, 2.2 Hz, 1H), 7.47 - 7.42 (m, 6H), 7.31 - 7.20 (m, 9H), 7.07 (d, J = 8.9 Hz, 1H), 4.47 - 4.39 (m, 2H), 4.14 (dt, J = 11.2, 5.9 Hz, 3H), 3.37 - 3.25 (m, 2H), 2.50 (d, J = 5.4 Hz, 1H), 1.86 (p, J = 6.8 Hz, 2H), 1.48 (p, J = 7.1 Hz, 2H), 1.27 (d, J = 6.8 Hz, 20H), 0.88 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 43 H 53 NO7[M+Na] + 718.3714, found 718.3715.

[0703] (S)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-(trityloxy)propyl 3-nitro-4-(tetradecyloxy)benzoate 3a-24

[0704]

[0705] Following the procedure described in 3a-4, EPA and 2a-15 were treated to give the target compound as a yellow oil in 69% yield. 1 HNMR (400 MHz, cdcl3) δ 8.28 (t, J = 1.8 Hz, 1H), 8.06 (dt, J = 8.9, 1.7 Hz, 1H), 7.45 - 7.40 (m, 6H), 7.33 - 7.19 (m, 9H), 7.06 (d, J = 8.8 Hz, 1H), 5.36 (dddd, J = 16.3, 11.9, 7.3, 2.4 Hz, 11H), 4.59 (dd, J = 11.9, 3.8 Hz, 1H), 4.50 (dd, J = 11.8, 6.4 Hz, 1H), 4.16 (t, J = 6.4 Hz, 2H), 3.37 - 3.24 (m, 2H), 2.79 (dt, J = 19.3, 5.7 Hz, 8H), 2.36 (t, J = 7.6 Hz, 2H), 2.09 (dt, J = 19.3, 7.2 Hz, 4H), 1.86 (p, J = 6.7 Hz, 2H), 1.70 (q, J = 7.5 Hz, 2H), 1.48 (p, J = 7.1 Hz, 2H), 1.26 (s, 20H), 0.96 (td, J = 7.5, 1.2 Hz, 3H), 0.90 - 0.84 (m, 3H). HRMS (ESI): calcd for C 63 H 81 NO8[M+Na] + 1002.5854, found 1002.5857.

[0706] (S)-3-hydroxy-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 3-nitro-4-(tetradecyloxy)benzoate 4a-24

[0707]

[0708] Following the procedure described in 4a-1, 3a-24 was treated to give the target compound as a yellow oil in 69% yield.1 H NMR (400 MHz, cdcl3) δ 8.45 (d, J = 2.2 Hz, 1H), 8.16 (dd, J = 8.8, 2.2 Hz, 1H), 7.10 (dd, J = 8.9, 1.6 Hz, 1H), 5.43 - 5.27 (m, 10H), 5.24 (dq, J = 5.9, 4.8 Hz, 1H), 4.57 (dd, J = 11.9, 4.3 Hz, 1H), 4.49 (dd, J = 11.9, 5.9 Hz, 1H), 4.16 (t, J = 6.4 Hz, 2H), 3.84 (dd, J = 20.6, 5.0 Hz, 2H), 2.87 - 2.75 (m, 8H), 2.42 - 2.33 (m, 2H), 2.16 - 2.01 (m, 4H), 1.85 (dt, J = 14.5, 6.6 Hz, 2H), 1.72 (p, J = 7.5 Hz, 2H), 1.48 (p, J = 7.0 Hz, 2H), 1.26 (s, 20H), 0.97 (t, J = 7.5 Hz, 3H), 0.92 - 0.84 (m, 3H). HRMS (ESI): calcd for C 44 H 67 NO8[M+Na] + 760.4759, found 760.4756.

[0709] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 3-nitro-4-(tetradecyloxy)benzoate 5a-24

[0710]

[0711] Following the procedure described in 5a-1, 4a-24 was treated to give the target compound as a colorless transparent oil in 49% yield. 1HNMR (400 MHz, cdcl3) δ 8.48 (d, J = 2.1 Hz, 1H), 8.19 (d, J = 8.5 Hz, 1H), 7.10 (d, J = 8.9 Hz, 1H), 5.56 (d, J = 8.4 Hz, 1H), 5.36 (dq, J = 10.5, 4.6 Hz, 10H), 4.67 - 4.55 (m, 1H), 4.48 (dd, J = 11.7, 4.6 Hz, 1H), 4.38 (dd, J = 11.5, 5.5 Hz, 1H), 4.30 (d, J = 8.8 Hz, 1H), 4.25 (d, J = 5.4 Hz, 2H), 4.16 (t, J = 6.4 Hz, 2H), 4.10 (s, 2H), 2.81 (dt, J = 11.5, 5.3 Hz, 8H), 2.35 (t, J = 7.6 Hz, 2H), 2.09 (dt, J = 18.6, 7.2 Hz, 4H), 1.85 (p, J = 6.7 Hz, 2H), 1.70 (p, J = 7.6 Hz, 2H), 1.50 - 1.19 (m, 49H), 0.97 (t, J = 7.5 Hz, 3H), 0.87 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 60 H 97 N2O 14 P[M+Na] + 1123.6570, found 1123.6569.

[0712] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 3-nitro-4-(tetradecyloxy)benzoate 6a-24

[0713]

[0714] According to the method described in 6a-1, 5a-24 was treated to obtain the target compound, colorless transparent oil, yield 95%. 1HNMR (400 MHz, cdcl3) δ 8.50 (d, J = 2.1 Hz, 1H), 8.22 (d, J = 8.8 Hz, 1H), 7.11 (d, J = 8.9 Hz, 1H), 5.58 (d, J = 8.2 Hz, 1H), 5.37 (tq, J = 12.8, 5.6 Hz, 10H), 4.87 - 4.79 (m, 1H), 4.59 - 4.43 (m, 2H), 4.32 (dtd, J = 26.5, 11.4, 6.1 Hz, 4H), 4.16 (t, J = 6.5 Hz, 2H), 2.82 (dt, J = 16.2, 5.2 Hz, 8H), 2.38 (t, J = 7.7 Hz, 2H), 2.09 (dp, J = 14.9, 7.1 Hz, 4H), 1.85 (p, J = 6.7 Hz, 2H), 1.72 (p, J = 7.6 Hz, 2H), 1.53 - 1.40 (m, 29H), 1.27 (d, J = 9.1 Hz, 20H), 0.97 (t, J = 7.6 Hz, 3H), 0.88 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 60 H 97 N2O 15 P[M+Na] + 1139.6519, found 1139.6520.

[0715] O-(hydroxy((R)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-((3-nitro-4-(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serine A24

[0716]

[0717] Following the procedure described for Al l, 6a-24 was treated to give the target compound as a brown solid in 76% yield. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.55 - 8.49 (m, 1H), 8.20 (br dd, J = 1.6, 8.8 Hz, 1H), 7.72 (br s, 2H), 7.18 - 7.11 (m, 1H), 5.64 - 5.11 (m, 10H), 4.85 (br d, J = 2.8 Hz, 1H), 4.64 (br d, J = 2.8 Hz, 3H), 4.54 - 4.43 (m, 3H), 4.33 (br dd, J = 4.4, 11.6 Hz, 1H), 4.17 (br t, J = 6.4 Hz, 2H), 3.00 - 2.58 (m, 8H), 2.44 (br t, J = 7.6 Hz, 2H), 2.16 - 2.02 (m, 4H), 1.90 - 1.82 (m, 2H), 1.77 - 1.66 (m, 2H), 1.27 (s, 22H), 0.97 (t, J = 7.6 Hz, 3H), 0.89 (br t, J = 6.8 Hz, 3H). 13 CNMR (101 MHz, CDC13 and 20 μL TFA) δ = 175.94, 169.00, 165.23, 156.66, 138.98, 135.87, 132.06, 129.28, 128.58, 128.30, 128.01, 127.84, 127.61, 127.00, 120.56, 114.23, 73.30, 70.37, 64.12, 63.69, 63.08, 54.30, 33.51, 31.91, 29.69, 29.65, 29.48, 29.35, 29.21, 28.66, 25.67, 25.50, 22.67, 20.52, 14.18, 14.06. HRMS (ESI): calcd for C 47 H 73 N2O 13 P[M-H] - 903.4778, found 903.4780.

[0718] Synthesis of compound A25

[0719] methyl 5-nitro-2-(tetradecyloxy)benzoate 1a-17

[0720]

[0721] Methyl 2-hydroxy-5-nitrobenzoate and 1-bromotetradecane were treated according to the procedure described for 1a-1 to give the target compound as a white solid in 99% yield. 1H NMR (400 MHz, cdcl3) δ 8.69 (d, J = 2.9 Hz, 1H), 8.33 (dd, J = 9.2, 2.9 Hz, 1H), 7.03 (d, J = 9.3 Hz, 1H), 4.14 (t, J = 6.5 Hz, 2H), 3.92 (s, 3H), 1.92 - 1.82 (m, 2H), 1.54 - 1.44 (m, 2H), 1.41 - 1.21 (m, 20H), 0.87 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 22 H 35 NO5[M+H] + 394.2588, found 394.2596.

[0722] 5-nitro-2-(tetradecyloxy)benzoic acid 1a-18

[0723]

[0724] Following the procedure described in 1a-2, 1a-17 was treated to give the target compound as a white solid in 96% yield. 1 H NMR (400 MHz, cdcl3) δ 9.04 (d, J = 2.9 Hz, 1H), 8.43 (dd, J = 9.2, 2.9 Hz, 1H), 7.16 (d, J = 9.2 Hz, 1H), 4.35 (t, J = 6.6 Hz, 2H), 2.01 - 1.90 (m, 2H), 1.59 - 1.44 (m, 2H), 1.25 (s, 20H), 0.91 - 0.83 (m, 3H). HRMS (ESI): calcd for C 21 H 33 NO5[M+H] + 380.2431, found 380.2439.

[0725] (S)-2-hydroxy-3-(trityloxy)propyl 5-nitro-2-(tetradecyloxy)benzoate2a-16

[0726]

[0727] Following the procedure described in 2a-1, 1a-18 and 1a were treated to give the target compound as a pale yellow oil in 69% yield. 1HNMR (400 MHz, cdcl3) δ 8.62 (d, J = 2.9 Hz, 1H), 8.34 (dd, J = 9.2, 2.9 Hz, 1H), 7.44 (dd, J = 7.7, 1.8 Hz, 6H), 7.29 (dd, J = 8.4, 6.5 Hz, 6H), 7.25 - 7.19 (m, 3H), 7.02 (d, J = 9.2 Hz, 1H), 4.49 - 4.39 (m, 2H), 4.16 - 4.06 (m, 3H), 3.31 (d, J = 5.2 Hz, 2H), 1.81 (p, J = 6.8 Hz, 2H), 1.44 (p, J = 7.0 Hz, 2H), 1.26 (d, J = 6.7 Hz, 20H), 0.88 (t, J = 6.7 Hz, 3H). HRMS (ESI): calcd for C 43 H 53 NO7[M+Na] + 718.3714, found 718.3711.

[0728] (S)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-(trityloxy)propyl 5-nitro-2-(tetradecyloxy)benzoate 3a-25

[0729]

[0730] Following the procedure described in 3a-4, EPA and 2a-16 were treated to give the target compound as a yellow oil in 69% yield. 1HNMR(400MHz,cdcl3)δ8.54(d,J=2.9Hz,1H),8.32(dd,J=9.2,2.9Hz,1H),7.45–7.38(m,6H),7.27(d,J=7.0Hz,4H),7.25(d,J=1 .2Hz,2H),7.23–7.16(m,1H),7.00(d,J=9.3Hz,1H),5.44–5.24(m,11H),4.60(dd,J=11.8,3.8Hz,1H),4.50(dd,J=11.8,6.5Hz, 1H),4.07(t,J=6.7Hz,2H),3.32(qd,J=10.0,5.0Hz,2H),2.86–2.72(m,8H),2.37(td,J=7.4,1.9Hz,2H),2.14–2.00(m,4H),1.7 5(dp,J=30.4,7.0Hz,4H),1.43(p,J=7.0Hz,2H),1.35–1.18(m,20H),0.95(t,J=7.5Hz,3H),0.90–0.84(m,3H).HRMS(ESI):calcd for C 63 H 81 NO8[M+Na] + 1002.5854,found 1002.5855.

[0731] (S)-3-hydroxy-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 5-nitro-2-(tetradecyloxy)benzoate 4a-25

[0732]

[0733] 3a-25 was treated according to the method described for 4a-1 to afford the title compound as a yellow oil in 94% yield. 1H NMR(400MHz,cdcl3)δ8.67(d,J=2.9Hz,1H),8.35(dd,J=9.2,2.9Hz,1H),7.04(d,J=9.3Hz,1H),5.45–5.25( m,10H),5.21(p,J=5.0Hz,1H),4.58(dd,J=11.9,4.5Hz,1H),4.49(dd,J=11.9,5.5Hz,1H),4.15(t,J=6.7Hz, 2H),3.83(s,2H),2.87–2.75(m,8H),2.43–2.36(m,2H),2.17–2.01(m,4H),1.87(p,J=6.8Hz,2H),1.73(p,J= 7.5Hz,2H),1.48(p,J=7.0Hz,2H),1.25(s,20H),0.97(t,J=7.5Hz,3H),0.91–0.84(m,3H).HRMS(ESI):calcd forC 44 H 67 NO8[M+Na] + 760.4759,found 760.4763.

[0734] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphanyl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 5-nitro-2-(tetradecyloxy)benzoate 5a-25

[0735]

[0736] 4a-25 was treated according to the method described for 5a-1 to afford the title compound as a light yellow oil in 69% yield. 1H NMR (400 MHz, cdcl3) δ 8.76 - 8.62 (m, 1H), 8.35 (dt, J = 9.2, 2.7 Hz, 1H), 7.05 (dd, J = 9.3, 1.9 Hz, 1H), 5.59 - 5.25 (m, 11H), 4.66 - 4.06 (m, 9H), 2.81 (dq, J = 18.8, 5.3 Hz, 8H), 2.37 (dt, J = 10.4, 7.6 Hz, 2H), 2.08 (ddt, J = 15.3, 7.5, 5.8 Hz, 4H), 1.88 (p, J = 6.9 Hz, 2H), 1.72 (p, J = 7.4 Hz, 4H), 1.52 - 1.24 (m, 49H), 0.97 (t, J = 7.6 Hz, 3H), 0.88 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 60 H 97 N2O 14 P[M+Na] + 1123.657, found 1123.6556.

[0737] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 5-nitro-2-(tetradecyloxy)benzoate 6a-25

[0738]

[0739] Following the procedure described in 6a-1, 5a-25 was treated to give the target compound as a light yellow oil in 35% yield. 1H NMR (400 MHz, cdcl3) δ 8.76 - 8.62 (m, 1H), 8.35 (dt, J = 9.2, 2.7 Hz, 1H), 7.05 (dd, J = 9.3, 1.9 Hz, 1H), 5.60 - 5.25 (m, 11H), 4.61 (ddd, J = 12.0, 6.1, 4.0 Hz, 1H), 4.56 - 4.06 (m, 8H), 2.81 (dq, J = 18.8, 5.3 Hz, 8H), 2.37 (dt, J = 10.4, 7.6 Hz, 2H), 2.08 (ddt, J = 15.3, 7.5, 5.8 Hz, 4H), 1.88 (p, J = 6.9 Hz, 2H), 1.72 (p, J = 7.4 Hz, 4H), 1.53 - 1.40 (m, 33H), 1.40 - 1.20 (m, 16H), 0.97 (t, J = 7.6 Hz, 3H), 0.88 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 60 H 97 N2O 15 P[M+Na] + 1139.6519, found 1139.6516.

[0740] O-(hydroxy((R)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-((5-nitro-2-(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serineA25

[0741]

[0742] Following the procedure described in A11, 6a-25 was treated to give the target compound as a brown solid in 26% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 8.81 - 8.60 (m, 1H), 8.38 (br d, J = 8.8 Hz, 1H), 8.04 - 7.49 (m, 3H), 7.10 (br d, J = 8.8 Hz, 1H), 5.68 - 5.06 (m, 4H), 4.93 - 4.32 (m, 6H), 4.32 - 3.86 (m, 4H), 2.89 - 2.66 (m, 2H), 2.59 - 2.27 (m, 2H), 2.19 - 1.93 (m, 3H), 1.92 - 1.51 (m, 6H), 1.51 - 1.08 (m, 27H), 1.07 - 0.71 (m, 6H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 168.98, 164.47, 163.95, 139.89, 132.04, 129.79, 128.57, 128.28, 128.20, 128.10, 127.98, 127.83, 118.59, 70.30, 63.74, 63.10, 59.68, 53.99, 31.89, 29.63, 29.48, 29.33, 29.13, 28.46, 25.52, 22.65, 14.02. HRMS (ESI): calcd for C 47 H 73 N2O 13 P[M-H] - 903.4778, found 903.4819.

[0743] Synthesis of compound A26 of example 26

[0744] 2-(tetradecyloxy)pyrimidine-5-carboxylic acid 1a-19

[0745]

[0746] Dissolve 2-chloropyrimidine-5-carboxylic acid (0.50 g, 3.15 mmol, 1.00 eq.) and tetradecan-1-ol (1.35 g, 6.31 mmol, 2.00 eq.) in DMSO (20 mL) solution, and add cesium carbonate (3.08 g, 9.46 mmol, 3.00 eq.) and KI (0.03 g, 0.16 mmol, 0.05 eq.), stir at 120 °C for 18 h. Monitor the reaction by TLC, after the reaction is completed, first dilute the reaction solution with water (100 mL), then adjust the PH of the solution to 2 with 2N dilute hydrochloric acid (30 mL), extract with ethyl acetate (50 mL x 3), combine the organic layer, dry with anhydrous sodium sulfate, concentrate under reduced pressure to obtain a residue. The residue is swelled with petroleum ether (50 mL), dried by filtration to obtain compound 1a-19, light yellow solid, yield 25%. 1 H NMR (400 MHz, CDC13) δ 9.10 (s, 2H), 4.45 (t, J = 6.7 Hz, 2H), 1.83 (p, J = 6.9 Hz, 2H), 1.52 - 1.41 (m, 2H), 1.25 (s, 20H), 0.91 - 0.83 (m, 3H). HRMS (ESI): calcd for C 19 H 32 N2O3[M+H] +337.2488, found 337.2483.

[0747] (S)-2-hydroxy-3-(trityloxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 2a-17

[0748]

[0749] Following the procedure described in 2a-1, 1a-19 and 1a were treated to give the target compound as a white solid in 81% yield. 1 HNMR (400 MHz, CDC13) δ = 9.12 (s, 2H), 7.46-7.41 (m, 7H), 7.35-7.29 (m, 8H), 5.35 (q, J = 4.8 Hz, 1H), 4.50-4.44 (m, 3H), 3.96 (d, J = 4.8 Hz, 2H), 3.47 (d, J = 4.8 Hz, 2H), 1.88-1.82 (m, 2H), 1.52-1.47 (m, 2H), 1.42-1.28 (m, 20H), 1.17 (d, J = 6.4 Hz, 1H), 0.91-0.88 (m, 3H). HRMS (ESI): calcd for C 41 H 52 N2O5[M+Na] + 675.3768, found 675.3767.

[0750] (S)-2-(stearoyloxy)-3-(trityloxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 3a-26

[0751]

[0752] Following the procedure described in 3a-4, stearic acid and 2a-17 were treated to give the target compound as a yellow oil in 41% yield. 1HNMR (400 MHz, CDC13) δ = 8.95 (s, 2H), 7.51-7.43 (m, 6H), 7.36-7.27 (m, 9H), 5.45-5.36 (m, 1H), 4.66-4.60 (m, 1H), 4.58-4.51 (m, 1H), 4.48 (t, J = 6.7 Hz, 2H), 3.40-3.30 (m, 2H), 2.37 (t, J = 7.2 Hz, 2H), 1.92-1.82 (m, 2H), 1.70-1.62 (m, 2H), 1.55-1.47 (m, 2H), 1.30 (br s, 46H), 0.92 (t, J = 6.4 Hz, 6H). HRMS (ESI): calcd for C 59 H 86 N2O6[M+Na] + 941.6378, found 941.6385.

[0753] (S)-3-hydroxy-2-(stearoyloxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 4a-26

[0754]

[0755] Following the procedure described in 4a-1, 3a-26 was treated to give the target compound as a white solid in 58% yield. 1 HNMR (400 MHz, CDC13) δ = 9.05 (s, 2H), 5.29-5.21 (m, 1H), 4.63-4.56 (m, 1H), 4.55-4.48 (m, 1H), 4.45 (t, J = 6.8 Hz, 2H), 3.83 (t, J = 5.2 Hz, 2H), 2.36 (t, J = 7.6 Hz, 2H), 2.00 (t, J = 6.4 Hz, 1H), 1.89-1.79 (m, 2H), 1.67-1.61 (m, 2H), 1.52-1.44 (m, 2H), 1.26 (br s, 46H), 0.89 (t, J = 6.4 Hz, 6H). HRMS (ESI): calcd for C 40 H 72 N2O6[M+Na] + 699.5283, found 699.5263.

[0756] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 2-(tetradecyloxy)pyrimidine-5- carboxylate 5a-26

[0757]

[0758] According to the method described in 5a-1, 4a-26 was treated to obtain the target compound, colorless transparent oil, yield 16%. 1 HNMR (400 MHz, CDC13) δ = 9.04 (s, 2H), 5.52-5.43 (m, 1H), 5.39-5.26 (m, 1H), 4.65-4.57 (m, 1H), 4.50-4.41 (m, 3H), 4.36-4.15 (m, 2H), 4.09-3.89 (m, 3H), 2.38-2.29 (m, 2H), 1.92-1.76 (m, 2H), 1.66-1.59 (m, 2H), 1.49-1.45 (m, 18H), 1.42 (d, J = 7.1 Hz, 9H), 1.34-1.24 (m, 48H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 56 H 102 N3O 12 P[M+Na] + 1062.7093, found 1062.7088.

[0759] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-(stearoyloxy)propyl 2-(tetradecyloxy)pyrimidine-5- carboxylate 5a-26

[0760]

[0761] According to the method described in 6a-1, 5a-26 was treated to obtain the target compound, white solid, yield 29%. 1H NMR (400 MHz, CDC13) δ = 9.04 (d, J = 0.8 Hz, 2H), 5.56 - 5.46 (m, 1H), 5.43 - 5.32 (m, 1H), 4.64 - 4.59 (m, 1H), 4.47 - 4.32 (m, 5H), 4.27 - 4.14 (m, 3H), 2.37 - 2.32 (m, 2H), 1.89 - 1.78 (m, 2H), 1.59 (br s, 2H), 1.51 - 1.47 (m, 18H), 1.46 - 1.44 (m, 9H), 1.26 (br s, 48H), 0.88 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 56 H 102 N3O 13 P[M+Na] + 1078.7042, found 1078.7033.

[0762] O-(hydroxy((R)-2-(stearoyloxy)-3-((2-(tetradecyloxy)pyrimidine-5-carbonyl)oxy)propoxy)phosphoryl)-D-serine A26

[0763]

[0764] According to the method described in Al, 6a-26 was treated to obtain the target compound, white solid, yield 75%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 9.36 (s, 2H), 7.75 (br s, 3H), 5.38 (br s, 1H), 4.77 - 4.44 (m, 7H), 4.21 (br s, 2H), 2.38 (br t, J = 7.2 Hz, 2H), 1.94 - 1.82 (m, 2H), 1.58 (br s, 2H), 1.44 (br s, 2H), 1.27 (br s, 48H), 0.89 (br t, J = 6.0 Hz, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 176.11, 168.76, 161.59, 160.97, 118.21, 73.42, 69.89, 69.81, 64.60, 63.81, 63.55, 54.24, 54.18, 34.07, 31.91, 29.69, 29.63, 29.57, 29.40, 29.35, 29.09, 29.03, 28.04, 25.24, 22.66, 14.01. HRMS (ESI): calcd for C 43 H 78 N3O 11 P[M+H] + 844.5447, found 844.5443.

[0765] Synthesis of compound A27

[0766] (S)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-(trityloxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 3a-27

[0767]

[0768] Following the procedure described for 3a-4, EPA and 2a-17 were treated to give the target compound as a yellow oil in 61% yield. 1 HNMR (400 MHz, cdcl3) δ 8.90 (s, 2H), 7.44 - 7.40 (m, 6H), 7.32 - 7.26 (m, 6H), 7.26 - 7.21 (m, 3H), 5.45 - 5.24 (m, 11H), 4.60 (dd, J = 11.8, 3.8 Hz, 1H), 4.51 (dd, J = 11.8, 6.2 Hz, 1H), 4.44 (t, J = 6.7 Hz, 2H), 3.36 - 3.26 (m, 2H), 2.86 - 2.74 (m, 8H), 2.35 (dd, J = 8.1, 7.1 Hz, 2H), 2.14 - 2.01 (m, 4H), 1.88 - 1.78 (m, 2H), 1.70 (p, J = 7.5 Hz, 2H), 1.52 - 1.41 (m, 2H), 1.26 (s, 20H), 0.96 (t, J = 7.5 Hz, 3H), 0.92 - 0.84 (m, 3H). HRMS (ESI): calcd for C 61 H 80N2O6[M+Na] + 959.5909, found 959.5912.

[0769] (S)-3-hydroxy-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 4a-27

[0770]

[0771] Following the procedure described in 4a-1, 3a-27 was treated to give the target compound as a yellow oil in 40% yield. 1 H NMR (400 MHz, cdcl3) δ 9.03 (s, 2H), 5.45 - 5.26 (m, 10H), 5.24 (dq, J = 6.1, 4.9 Hz, 1H), 4.59 (dd, J = 11.9, 4.2 Hz, 1H), 4.49 (dd, J = 11.9, 6.0 Hz, 1H), 4.43 (t, J = 6.7 Hz, 2H), 3.81 (d, J = 4.5 Hz, 2H), 2.86 - 2.76 (m, 8H), 2.41 - 2.32 (m, 2H), 2.16 - 2.01 (m, 4H), 1.82 (p, J = 6.8 Hz, 2H), 1.71 (p, J = 7.5 Hz, 2H), 1.52 - 1.40 (m, 2H), 1.39 - 1.21 (m, 20H), 0.97 (t, J = 7.5 Hz, 3H), 0.90 - 0.85 (m, 3H). HRMS (ESI): calcd for C 42 H 66 N2O6[M+H] + 695.4994, found695.4996.

[0772] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 5a-27

[0773]

[0774] Treatment of 4a-27 according to the procedure described in 5a-1 gave the title compound as a pale yellow oil in 94% yield. 1 H NMR (400 MHz, cdcl3) δ 9.01 (d, J = 3.3 Hz, 2H), 5.50 (dd, J = 11.3, 7.9 Hz, 1H), 5.28 (tdd, J = 15.3, 10.6, 6.9 Hz, 10H), 4.76 (tt, J = 8.3, 4.1 Hz, 1H), 4.50 (dt, J = 11.8, 3.6 Hz, 1H), 4.42 - 4.11 (m, 7H), 2.79 - 2.69 (m, 8H), 2.30 (dt, J = 9.8, 7.7 Hz, 2H), 2.02 (tt, J = 13.9, 7.0 Hz, 4H), 1.75 (p, J = 6.8 Hz, 2H), 1.64 (pd, J = 7.4, 2.2 Hz, 2H), 1.47 - 1.34 (m, 27H), 1.31 - 1.14 (m, 22H), 0.89 (t, J = 7.5 Hz, 3H), 0.80 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 58 H 96 N3O 12 P[M+Na] + 1080.6624, found 1080.6622.

[0775] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 2-(tetradecyloxy)pyrimidine-5-carboxylate 6a-27

[0776]

[0777] Treatment of 5a-27 according to the procedure described in 6a-1 gave the title compound as a pale yellow oil in 16% yield. 1H NMR (400 MHz, cdcl3) δ 9.01 (d, J = 3.3 Hz, 2H), 5.50 (dd, J = 11.3, 7.9 Hz, 1H), 5.28 (tdd, J = 15.3, 10.6, 6.9 Hz, 10H), 4.76 (tt, J = 8.3, 4.1 Hz, 1H), 4.50 (dt, J = 11.8, 3.6 Hz, 1H), 4.39 (dt, J = 13.5, 6.2 Hz, 3H), 4.24 (dddd, J = 28.1, 11.5, 8.3, 3.6 Hz, 4H), 2.79 - 2.70 (m, 8H), 2.30 (dt, J = 9.8, 7.7 Hz, 2H), 2.02 (tt, J = 13.9, 7.0 Hz, 4H), 1.75 (p, J = 6.8 Hz, 2H), 1.64 (pd, J = 7.4, 2.2 Hz, 2H), 1.47 - 1.34 (m, 27H), 1.33 - 1.08 (m, 22H), 0.89 (t, J = 7.5 Hz, 3H), 0.80 (t, J = 6.6 Hz, 3H). HRMS (ESI): calcd for C 58 H 96 N3O 13 P[M+Na] + 1096.6573, found 1096.6571.

[0778] O-(hydroxy((R)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-((2-(tetradecyloxy)pyrimidine-5-carbonyl)oxy)propoxy)phosphoryl)-D-serine A27

[0779]

[0780] Following the procedure described for Al l, 6a-27 was treated to give the target compound as a dark brown oil in 46% yield. 1H NMR (400 MHz, CDC13 and 20 uL TFA) δ = 9.50 - 9.23 (m, 2H), 7.93 - 7.57 (m, 2H), 5.75 - 5.45 (m, 1H), 5.45 - 5.06 (m, 5H), 4.96 - 4.77 (m, 1H), 4.76 - 4.36 (m, 7H), 4.35 - 3.76 (m, 2H), 2.95 - 2.60 (m, 3H), 2.58 - 2.21 (m, 3H), 2.21 - 1.93 (m, 4H), 1.92 - 1.77 (m, 3H), 1.77 - 1.54 (m, 4H), 1.51 (br s, 26H), 1.03 - 0.84 (m, 6H). 13 C NMR (101 MHz, CDC13 and 20 uL TFA) δ = 175.79, 168.69, 161.76, 132.09, 129.32, 128.61, 128.33, 128.23, 128.02, 127.99, 127.85, 118.29, 73.17, 68.57, 65.16, 63.66, 62.92, 54.18, 31.92, 29.65, 29.61, 29.52, 29.35, 29.06, 28.06, 26.29, 25.51, 25.26, 22.68, 14.17, 14.05. HRMS (ESI): calcd for C 45 H 72 N3O 11 P[M+H] + 862.4977, found 862.4980.

[0781] Synthesis of compound A28 of Example 28

[0782] tert-butyl (E)-4-(3-(3,4,5-trimethoxyphenyl)acrylamido)benzoate 1a-20

[0783]

[0784] To a solution of 4-amino benzoic acid tert-butyl ester (0.49 g, 2.52 mmol, 1.20 eq.) and (E)-3-(3,4,5-trimethoxyphenyl)acrylic acid (0.50 g, 2.10 mmol, 1.00 eq.) in 50 mL of dichloromethane, the reaction system was placed in an ice bath, and DCC (0.87 g, 4.20 mmol, 2.00 eq.) was added and stirred for ten minutes. Then, DMAP (0.13 g, 1.05 mmol, 0.50 eq.) was added, and the mixture was stirred at 0 °C for 2 h. The reaction was detected by TLC plate, and after the reaction was completed, the reaction mixture was filtered through celite and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 4 / 1 ~ 1 / 2) to obtain compound 1a-20, light yellow solid, yield 49%. 1 H NMR (400 MHz, cdcl3) δ 8.00-7.94 (m, 3H), 7.74-7.62 (m, 3H), 6.72 (s, 2H), 6.52 (d, J = 15.4 Hz, 1H), 3.87 (s, 3H), 3.84 (s, 6H), 1.58 (s, 9H). HRMS (ESI): calcd for C 23 H 27 NO6[M+H] + 414.1911, found 414.1908.

[0785] (E)-4-(3-(3,4,5-trimethoxyphenyl)acrylamido)benzoic acid 1a-21

[0786]

[0787] According to the method described in 4a-1, 1a-20 was treated to obtain the target compound, light yellow solid, yield 82%. 1 H NMR (400 MHz, cdcl3) δ 7.99 (s, 1H), 7.98-7.94 (m, 2H), 7.73-7.63 (m, 3H), 6.72 (s, 2H), 6.52 (d, J = 15.4 Hz, 1H), 3.87 (s, 3H), 3.84 (s, 6H). HRMS (ESI): calcd for C 19 H 19 NO6[M+H] + 358.1285, found 358.1283.

[0788] (S)-2-hydroxy-3-(trityloxy)propyl (E)-4-(3-(3,4,5-trimethoxyphenyl)acrylamido)benzoate 2a-18

[0789]

[0790] Following the procedure described in 2a-1, 1a-21 and 1a were treated to give the title compound as a light yellow solid in 80% yield. 1 HNMR (400 MHz, cdcl3) δ 7.97 - 7.91 (m, 2H), 7.73 - 7.64 (m, 4H), 7.48 - 7.40 (m, 6H), 7.32 - 7.28 (m, 6H), 7.26 - 7.22 (m, 3H), 6.78 (s, 2H), 6.49 (dd, J = 15.4, 1.4 Hz, 1H), 4.48 - 4.39 (m, 2H), 4.12 (q, J = 7.1 Hz, 1H), 3.90 (d, J = 1.8 Hz, 11H). HRMS (ESI): calcd for C 41 H 39 NO8[M+Na] + 696.2568, found 696.2562.

[0791] (S)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-(trityloxy)propyl 4-((E)-3-(3,4,5-trimethoxyphenyl)acrylamido)benzoate 3a-28

[0792]

[0793] Following the procedure described in 3a-4, EPA and 2a-18 were treated to give the title compound as a light yellow oil in 47% yield. 1H NMR (400 MHz, cdcl3) δ 7.91 - 7.84 (m, 2H), 7.79 - 7.63 (m, 4H), 7.45 - 7.37 (m, 6H), 7.31 - 7.17 (m, 9H), 6.76 (s, 2H), 6.48 (dd, J = 15.4, 1.7 Hz, 1H), 5.49 - 5.22 (m, 11H), 4.59 - 4.39 (m, 2H), 3.90 - 3.85 (m, 12H), 3.41 - 3.20 (m, 2H), 2.84 - 2.72 (m, 8H), 2.34 (dd, J = 8.2, 6.9 Hz, 2H), 2.13 - 1.99 (m, 4H), 1.69 (p, J = 7.5 Hz, 2H), 0.94 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 61 H 67 NO9[M+Na] + 980.4708, found 980.4713.

[0794] (S)-3-hydroxy-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 4-((E)-3-(3,4,5-trimethoxyphenyl)acrylamido)benzoate 4a-28

[0795]

[0796] Following the procedure described in 4a-1, 3a-28 was treated to give the target compound as yellow oil in 59% yield. 1 H NMR (400 MHz, cdcl3) δ 8.07 - 8.02 (m, 2H), 7.75 - 7.66 (m, 3H), 7.48 (s, 1H), 6.78 (s, 2H), 6.46 (d, J = 15.4 Hz, 1H), 5.46 - 5.30 (m, 10H), 5.31 - 5.25 (m, 1H), 4.41 (t, J = 4.6 Hz, 2H), 4.24 (t, J = 6.6 Hz, 2H), 3.91 (s, 6H), 3.90 (s, 3H), 2.88 - 2.77 (m, 8H), 2.38 (t, J = 7.6 Hz, 2H), 2.18 - 1.98 (m, 4H), 1.73 (p, J = 7.5 Hz, 2H), 0.97 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 42 H 53 NO9[M+Na] +738.3613, found 738.3607.

[0797] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 4-((E)-3-(3,4,5-trimethoxyphenyl)acrylamido)benzoate5a-28

[0798]

[0799] Following the procedure described in 5a-1, 4a-28 was treated to give the target compound as a brown oil in 59% yield. 1 H NMR (400 MHz, cdcl3) δ 8.05 - 7.95 (m, 3H), 7.76 - 7.71 (m, 2H), 7.67 (d, J = 15.4 Hz, 1H), 6.76 (s, 2H), 6.51 (d, J = 15.3 Hz, 1H), 5.58 (dt, J = 17.1, 7.4 Hz, 1H), 5.44 - 5.23 (m, 10H), 4.83 (dt, J = 9.5, 4.8 Hz, 1H), 4.56 - 4.02 (m, 6H), 3.87 (d, J = 2.2 Hz, 9H), 2.87 - 2.72 (m, 8H), 2.40 - 2.30 (m, 2H), 2.15 - 2.00 (m, 4H), 1.75 - 1.65 (m, 2H), 1.49 - 1.42 (m, 27H), 0.95 (td, J = 7.4, 1.4 Hz, 3H). HRMS (ESI): calcd for C 58 H 83 N2O 15 P[M+Na] + 1101.5423, found 1101.5421.

[0800] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propyl 4-((E)-3-(3,4,5-trimethoxyphenyl)acrylamido)benzoate6a-28

[0801]

[0802] Following the procedure described in 6a-1, 5a-28 was treated to give the target compound as a brown oil in 33% yield. 1 H NMR (400 MHz, cdcl3) δ 8.05 - 7.97 (m, 3H), 7.76 - 7.71 (m, 2H), 7.67 (d, J = 15.4 Hz, 1H), 6.76 (s, 2H), 6.51 (d, J = 15.3 Hz, 1H), 5.58 (dt, J = 17.1, 7.4 Hz, 1H), 5.44 - 5.26 (m, 10H), 4.83 (dt, J = 9.4, 4.9 Hz, 1H), 4.57 - 4.29 (m, 4H), 4.28 - 4.02 (m, 2H), 3.87 (d, J = 2.2 Hz, 9H), 2.87 - 2.73 (m, 8H), 2.40 - 2.30 (m, 2H), 2.15 - 2.00 (m, 4H), 1.75 - 1.65 (m, 2H), 1.49 - 1.42 (m, 27H), 0.95 (td, J = 7.4, 1.4 Hz, 3H). HRMS (ESI): calcd for C 58 H 83 N2O 16 P[M+Na] + 1117.5372, found 1117.5264.

[0803] O-(hydroxy((R)-2-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-((4-((E)-3-(3,4,5-trimethoxyphenyl)acrylamido)benzoyl)oxy)propoxy)phosphoryl)-D-serine A28

[0804]

[0805] Following the procedure described in A11, 6a-28 was treated to give the target compound as a black oil in 42% yield. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.98 - 7.55 (m, 8H), 6.74 (br dd, J = 2.8, 10.4 Hz, 2H), 6.49 (dt, J = 2.0, 11.6 Hz, 1H), 5.73 - 5.09 (m, 4H), 5.01 (br s, 1H), 4.68 - 4.26 (m, 7H), 3.94 - 3.81 (m, 11H), 2.90 - 2.64 (m, 2H), 2.49 - 2.26 (m, 3H), 2.19 - 1.98 (m, 4H), 1.68 (br s, 7H), 1.39 - 1.20 (m, 6H), 0.97 - 0.85 (m, 3H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 184.64, 172.54, 167.02, 166.81, 153.11, 141.95, 141.93, 139.56, 135.75, 135.18, 134.28, 131.06, 131.02, 130.92, 129.89, 128.31, 127.97, 120.44, 120.18, 118.42, 118.36, 105.54, 69.58, 63.68, 63.64, 61.17, 55.98, 29.71, 25.54, 25.47, 20.63, 14.10. HRMS (ESI): calcd for C 45 H 59 N2O 14 P[M+H] + 883.3777, found 883.3774.

[0806] Synthesis of compound A29 of Example 29

[0807] (S)-2-hydroxy-3-(trityloxy)propyl (5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 2a-19

[0808]

[0809] Following the procedure described in 2a-1, EPA and la were treated to give the target compound as a light brown oil in 33% yield. 1HNMR (400 MHz, cdcl3) δ 7.42 (d, J = 7.7 Hz, 6H), 7.27 (dt, J = 24.2, 7.2 Hz, 9H), 5.45 - 5.27 (m, 10H), 4.24 - 4.14 (m, 2H), 4.00 (t, J = 5.4 Hz, 1H), 3.25 - 3.17 (m, 2H), 2.83 (dt, J = 13.0, 5.5 Hz, 8H), 2.30 (t, J = 7.6 Hz, 2H), 2.13 - 2.03 (m, 4H), 1.66 (p, J = 7.5 Hz, 2H), 0.97 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 42 H 50 O4[M+Na] + 641.3601, found 641.3609.

[0810] (S)-1-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-(trityloxy)propan-2-yl 3-nitrobenzoate 3a-29

[0811]

[0812] Following the procedure described in 3a-4, 3-nitrobenzoic acid and 2a-19 were treated to give the target compound as a light brown oil in 69% yield. 1 H NMR (400 MHz, cdcl3) δ 8.87 (d, J = 2.2 Hz, 1H), 8.42 (ddd, J = 22.2, 8.0, 1.9 Hz, 2H), 7.68 (t, J = 8.0 Hz, 1H), 7.42 (dd, J = 8.1, 1.9 Hz, 6H), 7.26 (dt, J = 10.4, 6.8 Hz, 9H), 5.54 (dq, J = 9.1, 4.8 Hz, 1H), 5.43 - 5.25 (m, 9H), 4.49 (dd, J = 12.1, 3.5 Hz, 1H), 4.40 (dd, J = 12.1, 6.7 Hz, 1H), 3.41 (d, J = 4.9 Hz, 2H), 2.79 (dt, J = 25.0, 6.1 Hz, 8H), 2.26 (t, J = 7.6 Hz, 2H), 2.11 - 2.01 (m, 4H), 1.63 (q, J = 7.5 Hz, 2H), 0.96 (td, J = 7.5, 1.2 Hz, 3H). HRMS (ESI): calcd for C 49 H 53 NO7[M+Na] +790.3714, found 790.3714.

[0813] (S)-1-hydroxy-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propan-2-yl 3-nitrobenzoate 4a-29

[0814]

[0815] Following the procedure described in 4a-1, 3a-32 was treated to give the target compound as a brown oil in 96% yield. 1 H NMR (400 MHz, cdcl3) δ 8.86 (s, 1H), 8.50 - 8.35 (m, 2H), 7.68 (t, J = 8.0 Hz, 1H), 5.44 - 5.25 (m, 11H), 4.49 (dd, J = 12.2, 4.2 Hz, 1H), 4.42 (dd, J = 12.2, 5.7 Hz, 1H), 3.90 (t, J = 5.6 Hz, 2H), 2.80 (dt, J = 18.3, 5.9 Hz, 8H), 2.36 (t, J = 7.6 Hz, 2H), 2.08 (dd, J = 15.2, 7.5 Hz, 4H), 1.70 (p, J = 7.5 Hz, 2H), 0.96 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 30 H 39 NO7[M+Na] + 548.2619, found 548.2617.

[0816] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propan-2-yl 3-nitrobenzoate 5a-29

[0817]

[0818] Following the procedure described in 5a-1, 4a-29 was treated to give the target compound as a light brown oil in 30% yield. 1H NMR (400 MHz, cdcl3) δ 8.86 (s, 1H), 8.45 - 8.42 (m, 1H), 8.38 (d, J = 7.8 Hz, 1H), 7.67 (t, J = 8.0 Hz, 1H), 5.51 - 5.26 (m, 11H), 4.49 (dt, J = 12.2, 3.6 Hz, 1H), 4.38 - 4.23 (m, 2H), 4.02 (ddt, J = 30.9, 12.5, 7.1 Hz, 4H), 2.79 (dt, J = 22.4, 5.9 Hz, 8H), 2.33 (t, J = 7.6 Hz, 2H), 2.06 (p, J = 7.6 Hz, 4H), 1.67 (p, J = 7.5 Hz, 2H), 1.48 - 1.34 (m, 27H), 0.96 (t, J = 7.6 Hz, 3H). HRMS (ESI): calcd for C 46 H 69 N2O 13 P[M+Na] + 911.4429, found 911.4422.

[0819] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propan-2-yl 3-nitrobenzoate 6a-29

[0820]

[0821] Following the procedure described in 6a-1, 5a-29 was treated to give the target compound as a light brown oil in 64% yield. 1H NMR (400 MHz, cdcl3) δ 8.85 (s, 1H), 8.47 - 8.36 (m, 2H), 7.67 (t, J = 8.0 Hz, 1H), 7.26 (s, OH), 5.56 - 5.24 (m, 11H), 4.49 (dt, J = 12.4, 4.7 Hz, 1H), 4.41 - 4.17 (m, 6H), 2.78 (dt, J = 20.7, 5.9 Hz, 8H), 2.33 (t, J = 7.7 Hz, 2H), 2.06 (dq, J = 11.8, 7.1 Hz, 4H), 1.67 (p, J = 7.6 Hz, 2H), 1.45 (dd, J = 12.0, 8.6 Hz, 27H), 0.95 (t, J = 7.6 Hz, 3H). HRMS (ESI): calcd for C 46 H 69 N2O 14 P[M+Na] + 927.4379, found 927.4377.

[0822] O-(hydroxy((R)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-2-((3-nitrobenzoyl)oxy)propoxy)phosphoryl)-D-serine A29

[0823]

[0824] Following the procedure described in A11, 6a-29 was treated to give the target compound as a brown oil in 32% yield. 1 H NMR (400 MHz, CDCl3 and 20 μL TFA) δ = 8.80 (br s, 1H), 8.46 - 8.32 (m, 2H), 7.78 - 7.64 (m, 3H), 5.95 - 4.75 (m, 8H), 4.68 - 4.19 (m, 8H), 2.96 - 2.58 (m, 4H), 2.56 - 2.21 (m, 4H), 2.15 - 1.94 (m, 4H), 1.66 (br s, 6H), 0.96 (br dd, J = 7.2, 14.8 Hz, 3H). 13C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.35, 168.86, 164.60, 148.09, 135.72, 132.07, 129.31, 128.59, 128.31, 128.14, 127.97, 127.91, 126.98, 71.48, 65.21, 62.59, 54.23, 33.54, 29.70, 26.26, 25.48, 24.48, 20.50, 14.14. HRMS (ESI): calcd for C 33 H 45 N2O 12 P[M+H] + 691.2637, found 691.2615.

[0825] Synthesis of compound A30 of example 30

[0826] tert-butyl (10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)malonate 1a-22

[0827]

[0828] Tetraethylammonium bromide (0.50 g, 2.16 mmol, 1.00 eq.) and compound 1a-22 (0.50 g, 1.16 mmol, 0.50 eq.) were dissolved in 50 mL of methanol, and the reaction system was placed in an ice bath, and DCM (0.61 g, 2.96 mmol, 2.00 eq.) was added and stirred for ten minutes. Then, DMAP (0.09 g, 0.74 mmol, 0.50 eq.) was added, and the mixture was stirred at 0 °C for 2 h. TLC plate was used to detect whether the reaction was complete, and after the reaction was completed, the reaction mixture was filtered through diatomite, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 15 / 1 ~ 5 / 1) to obtain compound A30, orange red oil, yield 89%. 1 H NMR (400 MHz, CDC13) δ 4.12 (t, J = 6.7 Hz, 2H), 3.98 (d, J = 1.2 Hz, 6H), 3.27 (s, 2H), 2.44 (t, J = 7.4 Hz, 2H), 2.01 (s, 3H), 1.67 - 1.61 (m, 2H), 1.46 (s, 11H), 1.41 - 1.22 (m, 12H). HRMS (ESI): calcd for C 26 H 40 O8[M+Na]+ 503.2615, found 503.2617.

[0829] 3-((10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)oxy)-3-oxopropanoic acid 1a-23

[0830]

[0831] Following the procedure described in 4a-1, 1a-22 was treated to give the target compound as an orange red oil in 88% yield. 1 H NMR (400 MHz, cdcl3) δ 4.18 (t, J = 6.7 Hz, 2H), 3.98 (d, J = 1.3 Hz, 6H), 3.44 (s, 2H), 2.44 (t, J = 7.4 Hz, 2H), 2.01 (s, 3H), 1.65 (p, J = 6.7 Hz, 2H), 1.44 - 1.23 (m, 12H), 1.10 (s, 2H). HRMS (ESI): calcd for C 22 H 32 O8[M+H] + 425.217, found 425.2166.

[0832] 10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl((S)-1-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)-3-(trityloxy)propan-2-yl)malonate 3a-30

[0833]

[0834] Following the procedure described in 3a-4, 1a-23 and 2a-19 were treated to give the target compound as a red oil in 70% yield. 1H NMR(400MHz,cdcl3)δ7.44–7.37(m,6H),7.33–7.28(m,6H),7.26–7.21(m,3H),5.46–5.22(m,11H),4.34(dd,J =11.9,3.8Hz,1H),4.25(dd,J=11.9,6.6Hz,1H),4.10(td,J=6.9,2.4Hz,2H),3.99(d,J=0.7Hz,6H),3.40(d,J =0.8Hz,2H),3.31–3.22(m,2H),2.89–2.75(m,8H),2.44(t,J=7.4Hz,2H),2.25(t,J=7.6Hz,2H),2.12–2.03(m ,4H),2.01(s,3H),1.63(dt,J=15.6,7.7Hz,4H),1.41–1.22(m,14H),0.97(t,J=7.5Hz,3H).HRMS(ESI):calcd for C 64 H 80 O 11 [M+Na] + 1047.5593,found 1047.5591.

[0835] 10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl((S)-1-hydroxy-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propan-2-yl)malonate 4a-30

[0836]

[0837] 3a-30 was treated according to the method described for 4a-1 to obtain the title compound as a red oil in 51% yield. 1H NMR (400 MHz, cdcl3) δ 5.45 - 5.27 (m, 10H), 5.17 (tt, J = 5.8, 4.3 Hz, 1H), 4.36 - 4.22 (m, 2H), 4.14 (td, J = 6.8, 1.3 Hz, 2H), 3.98 (d, J = 1.1 Hz, 6H), 3.84 - 3.68 (m, 2H), 3.51 - 3.36 (m, 2H), 2.90 - 2.76 (m, 8H), 2.47 - 2.41 (m, 2H), 2.34 (t, J = 7.6 Hz, 2H), 2.16 - 2.04 (m, 4H), 2.01 (s, 3H), 1.75 - 1.61 (m, 4H), 1.43 - 1.22 (m, 14H), 0.97 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 45 H 66 O 11 [M+Na] + 805.4497, found 805.4490.

[0838] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propan-2-yl(10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)malonate 5a-30

[0839]

[0840] Following the procedure described for 5a-1, 4a-30 was treated to give the target compound as a brownish red oil in 40% yield. 1H NMR (400 MHz, cdcl3) δ 5.55 - 5.26 (m, 10H), 5.21 (tt, J = 10.7, 5.3 Hz, 1H), 4.44 - 4.22 (m, 2H), 4.18 (p, J = 6.0 Hz, 1H), 4.12 (t, J = 6.9 Hz, 2H), 4.10 - 4.01 (m, 2H), 3.99 (d, J = 0.9 Hz, 6H), 3.97 - 3.77 (m, 2H), 3.39 (dd, J = 6.5, 2.3 Hz, 2H), 2.87 - 2.78 (m, 8H), 2.44 (t, J = 7.4 Hz, 2H), 2.38 - 2.29 (m, 2H), 2.15 - 2.03 (m, 4H), 2.01 (s, 3H), 1.75 - 1.57 (m, 4H), 1.46 (s, 9H), 1.44 (d, J = 1.3 Hz, 9H), 1.40 (d, J = 1.7 Hz, 9H), 1.37 - 1.23 (m, 14H), 0.97 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 61 H 96 NO 17 P[M+Na] + 1168.6308, found 1168.6303.

[0841] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propan-2-yl(10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)malonate 6a-30

[0842]

[0843] Following the procedure described for 6a-1, 5a-30 was treated to give the target compound as a brownish red oil in 40% yield. 1H NMR (400 MHz, cdcl3) δ 5.58 - 5.48 (m, 1H), 5.45 - 5.21 (m, 10H), 4.34 (ddd, J = 12.1, 8.7, 5.3 Hz, 3H), 4.28 - 4.04 (m, 6H), 4.01 - 3.96 (m, 6H), 3.42 - 3.40 (m, 2H), 2.86 - 2.77 (m, 8H), 2.43 (t, J = 7.4 Hz, 2H), 2.33 (td, J = 7.6, 2.2 Hz, 2H), 2.14 - 2.02 (m, 4H), 2.00 (s, 3H), 1.75 - 1.58 (m, 4H), 1.51 - 1.46 (m, 18H), 1.44 (s, 9H), 1.44 - 1.22 (m, 14H), 0.96 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 61 H 96 NO 18 P[M+Na] + 1184.6257, found 1184.6253.

[0844] O-(((R)-2-((3-((10-(4,5-dimethoxy-2-methyl-3,6-dioxocyclohexa-1,4-dien-1-yl)decyl)oxy)-3-oxopropanoyl)oxy)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)

[0845] oxy)propoxy)(hydroxy)phosphoryl)-D-serine A30

[0846]

[0847] According to the method described in A11, 6a-30 was treated to give the target compound, black solid, yield 15%. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.10 - 7.45 (m, 3H), 5.36 (br dd, J = 4.6, 9.1 Hz, 4H), 4.77 - 4.41 (m, 3H), 4.40 - 4.05 (m, 7H), 4.04 - 3.78 (m, 6H), 3.52 (br s, 2H), 2.93 - 2.53 (m, 4H), 2.52 - 2.31 (m, 4H), 2.22 - 2.06 (m, 3H), 2.03 (s, 3H), 1.91 - 1.51 (m, 7H), 1.28 (br s, 19H), 1.01 - 0.86 (m, 3H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 185.46, 184.87, 171.34, 170.06, 168.33, 167.13, 144.23, 144.15, 143.63, 139.27, 132.06, 129.14, 128.59, 128.43, 128.29, 128.14, 128.02, 127.84, 126.99, 68.43, 66.94, 66.89, 61.22, 60.90, 60.86, 40.57, 40.19, 33.37, 29.74, 29.33, 29.28, 29.10, 29.07, 28.64, 28.08, 26.40, 25.57, 20.53, 14.19, 11.96, 11.86. HRMS (ESI): calcd for C 48 H 72 NO 16 P[M + H] + 950.4661, found 950.4655.

[0848] Synthesis of compound B1 of Example 31

[0849] (S)-3-(benzyloxy)-2-((9Z,12Z)-octadeca-9,12-dienamido)propyl(9Z,12Z)-octadeca-9,12-dienoate 3b-1

[0850]

[0851] (R)-2-amino-3-benzyloxy-1-propanol (1b) and linoleic acid were treated according to the procedure described for 3a-1 to give the target compound as a light yellow oil in 100% yield. 1H NMR(400MHz,Chloroform-d)δ7.38–7.32(m,2H),7.32–7.27(m,3H),5.83(d,J=8.6Hz,1H),5.44–5.25(m,8H),4 .50(s,2H),4.36(dqd,J=9.4,6.0,3.6Hz,1H),4.23(dd,J=11.0,6.3Hz,1H),4.13(dd,J=11.1,6.1Hz,1H),3.59 (dd,J=9.6,3.3Hz,1H),3.47(dd,J=9.5,4.8Hz,1H),2.77(t,J=6.4Hz,4H),2.26(t,J=7.6Hz,2H),2.18–2.12(m ,2H),2.04(q,J=6.8Hz,8H),1.59(h,J=7.4Hz,4H),1.41–1.22(m,28H),0.88(t,J=6.8Hz,6H).HRMS(ESI):calcd for C 46 H 75 NO4[M+Na] + 728.5588,found728.5586.

[0852] (S)-3-hydroxy-2-((9Z,12Z)-octadeca-9,12-dienamido)propyl(9Z,12Z)-octadeca-9,12-dienoate 4b-1

[0853]

[0854] Under nitrogen at -40°C, 3b-1 (0.78 g, 1.11 mmol, 1.00 eq.) was dissolved in dichloromethane (30 mL), followed by the addition of boron trichloride (1.00 M, 3.3 mL, 3.00 eq.) and stirring for 30 min. The mixture was then stirred at -10°C for an additional 30 min under nitrogen. The reaction mixture was quenched by the addition of saturated ammonium chloride solution (20 mL) at -10°C and then extracted with dichloromethane (50 mL × 3). The combined organic layers were washed with brine (50 mL × 3), dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel chromatography (PE / EA = 3 / 1 to 1 / 2) to obtain compound 3a-1 as a light yellow oil in an 89% yield. 1H NMR(400MHz,Chloroform-d)δ5.94(d,J=7.3Hz,1H),5.41–5.25(m,8H),4.25–4.11(m,3H),3.62(qd,J=12.6,12.1,3.7Hz,2H),2.75(t,J=6.4Hz,4H) ,2.32(t,J=7.6Hz,2H),2.21–2.12(m,2H),2.03(q,J=6.8Hz,8H),1.61(p,J=7.3Hz,4H),1.40–1.19(m,28H),0.87(t,J=6.8Hz,6H).HRMS(ESI):calcd for C 39 H 69 NO4[M+Na] + 683.5119, found 683.5127.

[0855] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphanyl)oxy)-2-((9Z,12Z)-octadeca-9,12-dienamido)propyl(9Z,12Z)-octadeca-9,12-dienoate 5b-1

[0856]

[0857] 4b-1 was treated according to the method described for 5a-1 to obtain the title compound as a light yellow oil in 65% yield. 1 H NMR(400MHz,Chloroform-d)δ6.09–5.93(m,1H),5.46–5.26(m,8H),4.32(ddt,J=15.1 ,11.7,4.6Hz,2H),4.24–3.66(m,6H),2.76(t,J=6.7Hz,4H),2.30(t,J=7.6Hz,2H),2.1 8(q,J=6.8Hz,2H),2.04(q,J=6.8Hz,8H),1.60(d,J=14.1Hz,4H),1.45(d,J=8.2Hz,18 H),1.39(d,J=2.2Hz,9H),1.39–1.22(m,28H),0.88(t,J=6.9Hz,6H).HRMS(ESI):calcd forC 55 H 99 N2O 10 P[M+Na]+ 1001.693, found 1001.684.

[0858] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-((9Z,12Z)-octadeca-9,12-dienamido)propyl(9Z,12Z)-octadeca-9,12-dienoate 6b-1

[0859]

[0860] Following the procedure described for 6a-1, 5b-1 was treated to give the target compound as a light yellow oil in 60% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.09 - 5.93 (m, 1H), 5.46 - 5.26 (m, 8H), 4.32 (ddt, J = 15.1, 11.7, 4.6 Hz, 2H), 4.24 - 3.66 (m, 6H), 2.76 (t, J = 6.7 Hz, 4H), 2.30 (t, J = 7.6 Hz, 2H), 2.18 (q, J = 6.8 Hz, 2H), 2.04 (q, J = 6.8 Hz, 8H), 1.60 (d, J = 14.1 Hz, 4H), 1.45 (d, J = 8.2 Hz, 18H), 1.39 (d, J = 2.2 Hz, 9H), 1.39 - 1.22 (m, 28H), 0.88 (t, J = 6.9 Hz, 6H). HRMS (ESI): calcd for C 55 H 99 N2O 11 P[M+Na] + 1017.6879, found 1017.6883.

[0861] O-(hydroxy((R)-2-((9Z,12Z)-octadeca-9,12-dienamido)-3-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propoxy)phosphoryl)-D-serine B1

[0862]

[0863] Following the procedure described for A11, 6b-1 was treated to give the target compound as a yellow solid in 27% yield.1 H NMR (400 MHz, CDC13 and 20 uL TFA) d = 7.81 (br s, 3H), 7.27 (s, 1H), 5.61-5.20 (m, 3H), 5.06 (br d, J = 5.4 Hz, 1H), 4.45 (br s, 4H), 4.29-3.78 (m, 4H), 2.81-2.29 (m, 6H), 2.12-1.93 (m, 4H), 1.80-1.49 (m, 8H), 1.28 (br s, 32H), 0.97-0.82 (m, 7H). 13 C NMR (101 MHz, CDC13 and 20 uL TFA) d = 178.00, 175.78, 169.04, 157.64, 135.50, 134.16, 130.27, 129.94, 128.11, 123.34, 122.60, 64.87, 63.77, 62.62, 54.02, 49.36, 37.13, 33.94, 31.56, 29.58, 29.33, 29.10, 28.96, 28.79, 27.17, 24.65, 24.56, 22.55, 13.99. HRMS (ESI): calcd for C 42 H 75 N2O9P[M-H] - 781.5137, found 781.5112.

[0864] Synthesis of compound B2 of Example 32

[0865] (9Z,12Z)-N-((R)-2,3-dihydroxypropyl)octadeca-9,12-dienamide 1c

[0866]

[0867] Linoleic acid (1.00 g, 3.57 mmol, 1.00 eq.) was acylchlorinated with dichlorosulfide (3 mL) at 50 °C for 3 h, the residual dichlorosulfide was removed under vacuum, and the solution was set aside for later use. (R)-3-aminopropane-1,2-diol (0.36 g, 3.93 mmol, 1.10 eq.) was dissolved in tetrahydrofuran (20 mL), and the solution was adjusted to pH 10-11 by adding saturated sodium bicarbonate solution (10 mL) and stirring for 5 min. The acylchlorinated linoleic acid was added under ice bath, and the solution was stirred at room temperature for 2 h. The reaction was monitored by TLC, and after the reaction was completed, the solution was extracted with dichloromethane (30 mL x 3), and the combined organic layers were washed with saturated brine (30 mL x 3) and then dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 20 / 1-10 / 1) to obtain compound 1c as a brown oil in 55% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.13 (t, J = 6.3 Hz, 1H), 5.47-5.24 (m, 4H), 3.76 (p, J = 5.1 Hz, 1H), 3.61-3.50 (m, 2H), 3.40 (ddt, J = 14.3, 11.6, 7.2 Hz, 3H), 2.77 (t, J = 6.4 Hz, 2H), 2.22 (t, J = 7.6 Hz, 2H), 2.05 (q, J = 6.9 Hz, 4H), 1.63 (p, J = 7.1 Hz, 2H), 1.42-1.22 (m, 14H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 21 H 39 NO3[M+Na] + 376.2822, found 376.2821.

[0868] (9Z,12Z)-N-((R)-2-hydroxy-3-((4-methoxyphenyl)diphenylmethoxy)propyl)octadeca-9,12-dienamide 2c

[0869]

[0870] 1c (0.55 g, 1.98 mmol, 1.00 eq.), tetrabutylammonium chloride (0.23 g, 0.63 mmol, 0.32 eq.), and diisopropylamine (0.77 g, 5.93 mmol, 3.00 eq.) were dissolved in 20 mL of tetrahydrofuran. (Chloro(4-methoxyphenyl)methylene)diphenyl (0.92 g, 2.97 mmol, 1.50 eq.) was added and stirred at 50°C for 2 h. Completion of the reaction was confirmed by TLC. Upon completion, the reaction solution was quenched with 3N HCl (20 mL) and extracted with dichloromethane (30 mL × 3). The combined organic layers were washed with saturated brine (30 mL × 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column chromatography (PE / EA = 4 / 1 to 1 / 1) to obtain compound 2c as a brown oil in a 63% yield. 1 H NMR(400MHz,Chloroform-d)δ7.46–7.40(m,4H),7.34–7.18(m,11H),6.88–6.78(m,2H ),5.77(t,J=5.7Hz,1H),5.46–5.24(m,4H),4.11(q,J=7.1Hz,1H),3.78(s,3H),3.55(d dd,J=14.0,6.5,3.3Hz,1H),3.27–3.09(m,3H),2.77(t,J=6.4Hz,2H),2.12–2.01(m,6 H),1.54(p,J=7.3Hz,2H),1.40–1.21(m,14H),0.89(t,J=6.8Hz,3H).HRMS(ESI):calcd for C 41 H 55 NO4[M+Na] + 648.4023,found 648.4023.

[0871] (R)-1-((4-methoxyphenyl)diphenylmethoxy)-3-((9Z,12Z)-octadeca-9,12-dienamido)propan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 3c

[0872]

[0873] Linoleic acid (0.93 g, 1.49 mmol, 1.20 eq.) and 2c (0.77 g, 1.24 mmol, 1.00 eq.) were treated according to the method described in 3a-1 to obtain the title compound as a light brown oil in 87% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.44 - 7.38 (m, 4H), 7.32 - 7.16 (m, 8H), 6.84 - 6.79 (m, 2H), 5.69 (t, J = 5.7 Hz, 1H), 5.44 - 5.25 (m, 8H), 5.07 (dq, J = 6.7, 4.6 Hz, 1H), 3.77 (s, 3H), 3.57 (ddd, J = 14.2, 5.8, 4.4 Hz, 1H), 3.45 (ddd, J = 14.2, 6.9, 5.6 Hz, 1H), 3.29 - 3.15 (m, 2H), 2.81 - 2.66 (m, 4H), 2.33 (t, J = 7.6 Hz, 2H), 2.08 - 1.98 (m, 10H), 1.64 (q, J = 7.2 Hz, 2H), 1.51 (p, J = 7.4 Hz, 2H), 1.38 - 1.21 (m, 28H), 0.87 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 59 H 85 NO5[M+Na] + 910.6320, found 910.6320.

[0874] (R)-1-hydroxy-3-((9Z,12Z)-octadeca-9,12-dienamido)propan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 4c

[0875]

[0876] Following the procedure described in 4a-1, 4c was treated to give the target compound as yellow oil in 76% yield. 1 H NMR (400 MHz, Chloroform-d) δ 5.35 (tq, J = 8.5, 4.2, 3.2 Hz, 8H), 4.82 (p, J = 5.0 Hz, 0H), 4.14 (dd, J = 11.5, 5.2 Hz, 1H), 4.05 (dd, J = 11.5, 5.8 Hz, 1H), 3.62 - 3.51 (m, 2H), 2.76 (t, J = 6.5 Hz, 4H), 2.37 - 2.30 (m, 2H), 2.21 (q, J = 7.0 Hz, 2H), 2.04 (q, J = 6.9 Hz, 8H), 1.62 (q, J = 5.7, 4.9 Hz, 4H), 1.31 (d, J = 10.6 Hz, 28H), 0.88 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 39 H 69NO4[M+Na] + 638.5119, found 638.5121.

[0877] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-3-((9Z,12Z)-octadeca-9,12-dienamido)propan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 5c

[0878]

[0879] According to the method described in 5a-1, 4c was treated to obtain the target compound, yellow oil, yield 14%. 1 H NMR (400 MHz, Chloroform-d) δ 6.54 (s, 1H), 5.85 (d, J = 7.9 Hz, 1H), 5.43 - 5.25 (m, 7H), 4.42 (s, 1H), 4.26 (s, 1H), 4.12 (qd, J = 11.6, 5.6 Hz, 2H), 3.64 (ddd, J = 14.1, 6.6, 3.4 Hz, 1H), 3.30 - 3.21 (m, 1H), 2.77 (t, J = 6.6 Hz, 4H), 2.32 (t, J = 7.6 Hz, 2H), 2.25 - 2.15 (m, 2H), 2.05 (q, J = 7.0 Hz, 8H), 1.68 - 1.58 (m, 4H), 1.51 - 1.38 (m, 28H), 1.37 - 1.23 (m, 27H), 0.92 - 0.85 (m, 6H). HRMS (ESI): calcd for C 55 H 99 N2O 10 P[M+Mg] + 1002.6882, found 1002.6882.

[0880] (2R)-1-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-3-((9Z,12Z)-octadeca-9,12-dienamido)propan-2-yl(9Z,12Z)-octadeca-9,12-dienoate 6c

[0881]

[0882] Following the procedure described in 6a-1, 5c was treated to give the target compound as a yellow oil in 72% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.54 (s, 1H), 5.85 (d, J = 7.9 Hz, 1H), 5.43 - 5.25 (m, 8H), 4.42 (s, 1H), 4.26 (s, 1H), 4.12 (qd, J = 11.6, 5.6 Hz, 2H), 3.64 (ddd, J = 14.1, 6.6, 3.4 Hz, 1H), 3.30 - 3.21 (m, 1H), 2.77 (t, J = 6.6 Hz, 4H), 2.32 (t, J = 7.6 Hz, 2H), 2.25 - 2.15 (m, 2H), 2.05 (q, J = 7.0 Hz, 8H), 1.66 - 1.57 (m, 4H), 1.51 - 1.39 (m, 28H), 1.39 - 1.23 (m, 27H), 0.93 - 0.84 (m, 6H). HRMS (ESI): calcd for C 55 H 99 N2O 11 P[M+Na] + 1017.6879, found 1017.6888.

[0883] O-(hydroxy((R)-3-((9Z,12Z)-octadeca-9,12-dienamido)-2-(((9Z,12Z)-octadeca-9,12-dienoyl)oxy)propoxy)phosphoryl)-D-serine B2

[0884]

[0885] Following the procedure described in 6a-1, 5c was treated to give the target compound as a yellow oil in 72% yield. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.75 (br d, J = 4.4 Hz, 2H), 7.48 - 7.31 (m, 1H), 5.64 - 5.45 (m, 1H), 5.44 - 5.16 (m, 3H), 5.06 (br s, 1H), 4.58 (br s, 3H), 4.47 - 4.38 (m, 1H), 4.37 - 4.27 (m, 1H), 4.25 - 4.05 (m, 1H), 3.82 - 3.32 (m, 2H), 2.77 (br s, 1H), 2.36 (br d, J = 5.6 Hz, 4H), 2.13 - 1.90 (m, 4H), 1.79 - 1.47 (m, 8H), 1.28 (br s, 32H), 0.89 (br s, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 178.64, 176.19, 168.89, 135.51, 134.16, 130.25, 129.91, 128.09, 127.86, 123.15, 122.57, 73.20, 63.77, 63.27, 54.19, 41.31, 37.11, 33.60, 31.54, 31.49, 29.71, 29.32, 29.07, 28.97, 28.88, 28.76, 27.15, 22.54, 22.46, 13.93. HRMS (ESI): calcd for C 42 H 75 N2O9P[M-H] - 781.5137, found 781.5160.

[0886] Synthesis of compound B3 of example 33

[0887] (S)-3-(benzyloxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 3b-2

[0888]

[0889] Following the procedure described in 3a-1, (R)-2-amino-3-benzyloxy-1-propanol and EPA were treated to give the target compound as a yellow oil in 99% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.38 - 7.32 (m, 2H), 7.32 - 7.27 (m, 3H), 5.81 (d, J = 8.6 Hz, 1H), 5.45 - 5.25 (m, 29H), 4.50 (s, 2H), 4.40 - 4.31 (m, 1H), 4.23 (dd, J = 11.0, 6.2 Hz, 1H), 4.13 (dd, J = 11.0, 6.1 Hz, 1H), 3.59 (dd, J = 9.5, 3.3 Hz, 1H), 3.47 (dd, J = 9.5, 4.7 Hz, 1H), 2.88 - 2.74 (m, 16H), 2.27 (t, J = 7.6 Hz, 2H), 2.19 - 2.14 (m, 2H), 2.13 - 2.01 (m, 8H), 1.75 - 1.60 (m, 4H), 0.97 (t, J = 7.6 Hz, 6H). HRMS (ESI): calcd for C 50 H 71 NO4[M+H] + 750.5456, found 750.5452.

[0890] (S)-3-hydroxy-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 4b-2

[0891]

[0892] Following the procedure described for 4b-1, 3b-2 was treated to give the target compound as a yellow oil in 88% yield. 1 H NMR (400 MHz, Chloroform-d) δ 5.94 (d, J = 7.2 Hz, 1H), 5.46 - 5.25 (m, 20H), 4.28 - 4.15 (m, 3H), 3.66 (t, J = 14.2 Hz, 2H), 2.87 - 2.76 (m, 16H), 2.39 - 2.31 (m, 2H), 2.24 - 2.16 (m, 2H), 2.16 - 2.01 (m, 8H), 1.71 (pd, J = 7.6, 2.5 Hz, 4H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 43 H 65 NO4[M+H] + 660.4986, found 660.4995.

[0893] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 5b-2

[0894]

[0895] Following the procedure described in 5a-1, 4b-2 was treated to give the target compound as a light yellow oil in 34% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.52 - 6.33 (m, 1H), 5.49 (q, J = 9.3, 8.7 Hz, 1H), 5.45 - 5.25 (m, 20H), 4.47 - 3.98 (m, 8H), 2.88 - 2.75 (m, 16H), 2.33 (t, J = 7.8 Hz, 2H), 2.09 (p, J = 7.3 Hz, 6H), 1.76 - 1.66 (m, 4H), 1.52 - 1.42 (m, 27H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 59 H 95 N2O 10 P[M+Na] + 1045.6617, found 1045.6616.

[0896] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 6b-2

[0897]

[0898] Treatment of 5b-2 according to the method described in 6a-1 gave the target compound as a light yellow oil in 39% yield. 1 H NMR (400 MHz, Chloroform-d) δ 6.59 - 6.36 (m, 1H), 5.59 - 5.45 (m, 1H), 5.37 (hd, J = 7.7, 2.4 Hz, 20H), 4.47 - 3.97 (m, 8H), 2.90 - 2.76 (m, 16H), 2.33 (t, J = 7.7 Hz, 2H), 2.22 (ddt, J = 10.9, 5.5, 2.5 Hz, 2H), 2.15 - 2.01 (m, 8H), 1.70 (ddt, J = 14.8, 10.9, 5.1 Hz, 4H), 1.49 (d, J = 4.4 Hz, 18H), 1.45 (s, 9H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 59 H 95 N2O 11 P[M+Na] + 1061.6566, found 1061.6557.

[0899] O-(hydroxy((R)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)phosphoryl)-D-serine B3

[0900]

[0901] Treatment of 6b-2 according to the method described in A11 gave the target compound as a black oil in 24% yield. 1 H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 8.47 (br d, J = 2.6 Hz, 3H), 5.86 - 5.00 (m, 8H), 4.43 - 4.15 (m, 7H), 3.88 (br s, 4H), 3.51 - 3.31 (m, 2H), 2.89 - 2.69 (m, 4H), 2.27 - 1.88 (m, 14H), 1.85 - 1.42 (m, 15H), 1.32 - 1.13 (m, 6H), 0.95 - 0.80 (m, 6H). 13C NMR (400 MHz, DMSO-d6and 20 μL TFA) δ = 174.73, 172.86, 168.71, 131.87, 128.44, 128.32, 128.16, 128.13, 127.99, 127.23, 64.29, 63.68, 60.10, 58.94, 52.95, 33.37, 30.09, 29.34, 27.72, 25.50, 25.42, 24.76, 20.34, 14.35. HRMS (ESI): calcd for C 46 H 71 N2O9P[M-H] - 825.4824, found 825.4835.

[0902] Synthesis of compound B4 of Example 34

[0903] (S)-3-(benzyloxy)-2-(4-(tetradecyloxy)benzamido)propyl 4-(tetradecyloxy)benzoate 3b-3

[0904]

[0905] Following the procedure described in 3a-1, (R)-2-amino-3-benzyloxy-1-propanol and 1a-2 were treated to give the target compound as a white solid in 60% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.85 (d, J = 8.5 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 7.22 (dd, J = 16.3, 6.8 Hz, 5H), 6.81 (t, J = 8.6 Hz, 4H), 6.56 (d, J = 8.0 Hz, 1H), 5.32 - 5.23 (m, 1H), 4.69 - 4.35 (m, 4H), 4.14 - 3.31 (m, 6H), 1.73 (q, J = 7.5 Hz, 4H), 1.39 (q, J = 8.1, 7.6 Hz, 4H), 1.20 (d, J = 8.2 Hz, 40H), 0.81 (t, J = 6.6 Hz, 6H). HRMS (ESI): calcd for C 52 H 79 NO6[M+Na] + 836.5800, found 836.5802.

[0906] (S)-3-hydroxy-2-(4-(tetradecyloxy)benzamido)propyl 4-(tetradecyloxy)benzoate 4b-3

[0907]

[0908] Following the procedure described in 4b-1, 4b-3 was treated to give the target compound as a white solid in 57% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.98 (d, J = 8.3 Hz, 2H), 7.74 (d, J = 8.3 Hz, 2H), 6.90 (d, J = 8.4 Hz, 4H), 6.82 (d, J = 7.4 Hz, 1H), 4.64 - 4.41 (m, 3H), 3.99 (q, J = 7.1 Hz, 4H), 3.88 - 3.63 (m, 3H), 1.43 (d, J = 8.4 Hz, 4H), 1.37 - 1.19 (m, 44H), 0.87 (d, J = 13.4 Hz, 7H). HRMS (ESI): calcd for C 45 H 73 NO6[M+Na] + 746.5330, found 746.5331.

[0909] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-(4-(tetradecyloxy)benzamido)propyl 4-(tetradecyloxy)benzoate 5b-3

[0910]

[0911] Following the procedure described in 5a-1, 4b-3 was treated to give the target compound as a colorless transparent oil in 82% yield. 1HNMR (400 MHz, Chloroform-d) δ 7.96 (d, J = 8.0 Hz, 2H), 7.81 - 7.68 (m, 2H), 7.25 (s, 1H), 6.88 (d, J = 7.8 Hz, 4H), 4.69 - 4.24 (m, 5H), 4.18 - 3.85 (m, 7H), 3.47 (dt, J = 18.5, 7.0 Hz, 1H), 1.83 - 1.72 (m, 4H), 1.39 (d, J = 29.6 Hz, 27H), 1.26 (d, J = 10.4 Hz, 44H), 0.86 (d, J = 7.2 Hz, 7H). HRMS (ESI): calcd for C 61 H 103 N2O 12 P[M + Na] + 1109.7141, found 1109.7148.

[0912] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-(4-(tetradecyloxy)benzamido)propyl 4-(tetradecyloxy)benzoate 6b-3

[0913]

[0914] Following the procedure described for 6a-1, 5b-3 was treated to give the target compound as a colorless transparent oil in 82% yield. 1 HNMR (400 MHz, Chloroform-d) δ 7.97 (d, J = 8.4 Hz, 2H), 7.83 (d, J = 8.1 Hz, 2H), 6.90 (d, J = 8.3 Hz, 4H), 4.70 (s, 1H), 4.61 - 4.50 (m, 1H), 4.50 - 4.41 (m, 1H), 4.34 (d, J = 10.8 Hz, 3H), 4.31 - 4.17 (m, 3H), 3.99 (q, J = 7.0 Hz, 4H), 1.83 - 1.74 (m, 4H), 1.54 - 1.20 (m, 71H), 0.92 - 0.83 (m, 6H). HRMS (ESI): calcd for C 61 H 103 N2O 13 P[M + Na] + 1125.7090, found 1125.7092.

[0915] O-(hydroxy((R)-2-(4-(tetradecyloxy)benzamido)-3-((4-(tetradecyloxy)benzoyl)oxy)propoxy)phosphoryl)-D-serine B4

[0916]

[0917] According to the method described in A1, 6b-3 was treated to give the target compound, a light yellow solid, yield 58%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.91 (br d, J = 8.8 Hz, 2H), 7.77 (br s, 3H), 7.66 (br d, J = 8.0 Hz, 2H), 7.59 (br d, J = 7.6 Hz, 1H), 6.98-6.83 (m, 4H), 4.77 (br s, 1H), 4.57 (br s, 4H), 4.43 (br s, 1H), 4.28 (br s, 2H), 4.09-3.90 (m, 4H), 1.87-1.72 (m, 4H), 1.44 (br d, J = 4.8 Hz, 2H), 1.27 (s, 42H), 0.89 (t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 171.27, 168.85, 164.21, 163.42, 132.13, 129.37, 122.66, 119.86, 114.87, 114.56, 68.60, 65.42, 63.60, 63.38, 54.25, 50.19, 31.91, 29.68, 29.65, 29.53, 29.35, 28.96, 25.88, 22.67, 14.04. HRMS (ESI): calcd for C 48 H 79 N2O 11 P[M-H] - 889.5349, found 889.5337.

[0918] Synthesis of compound B5 in Example 35

[0919] (5E,8E,11E,14E,17E)-N-((R)-1-(benzyloxy)-3-hydroxypropan-2-yl)icosa-5,8,11,14,17-pentaenamide 2b-1

[0920]

[0921] Following the procedure described in 2a-1, EPA and 1b were treated to give the target compound as a brown oil in 88% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.72 (dd, J = 7.2, 2.2 Hz, 1H), 8.31 (ddd, J = 8.9, 4.3, 2.3 Hz, 1H), 7.40 (dd, J = 10.0, 8.7 Hz, 1H), 6.02 (d, J = 8.1 Hz, 1H), 5.37 (qd, J = 13.2, 11.3, 6.6 Hz, 10H), 4.60 - 4.44 (m, 2H), 4.43 - 4.31 (m, 1H), 3.86 - 3.67 (m, 2H), 2.88 - 2.75 (m, 8H), 2.27 - 2.19 (m, 2H), 2.15 - 2.01 (m, 4H), 1.70 (q, J = 6.9, 6.4 Hz, 2H), 1.03 - 0.84 (m, 3H). HRMS (ESI): calcd for C 30 H 43 NO3[M+Na] + 488.3135, found 488.3137.

[0922] (S)-3-(benzyloxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl 4-fluoro-3-nitrobenzoate 3b-4

[0923]

[0924] Following the procedure described in 3a-4, 4-fluoro-3-nitrobenzoic acid and 2b-1 were treated to give the target compound as a brown oil in 43% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.62 (dd, J = 7.2, 2.2 Hz, 1H), 8.22 (ddd, J = 8.8, 4.2, 2.2 Hz, 1H), 7.39 - 7.23 (m, 5H), 5.96 (d, J = 8.3 Hz, 1H), 5.46 - 5.25 (m, 10H), 4.59 - 4.38 (m, 5H), 3.69 (dd, J = 9.6, 2.6 Hz, 1H), 3.57 (dd, J = 9.6, 4.0 Hz, 1H), 2.90 - 2.73 (m, 8H), 2.23 - 2.15 (m, 2H), 2.12 - 2.03 (m, 4H), 1.75 - 1.65 (m, 2H), 0.97 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 37 H 45 FN2O6[M+H] + 633.3334, found 633.3332.

[0925] (S)-3-hydroxy-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl 4-fluoro-3-nitrobenzoate 4b-4

[0926]

[0927] Following the procedure described for 4b-1, 3b-4 was treated to give the target compound as a brown oil in 88% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.72 (dd, J = 7.2, 2.2 Hz, 1H), 8.31 (ddd, J = 8.9, 4.3, 2.3 Hz, 1H), 7.40 (dd, J = 10.0, 8.7 Hz, 1H), 6.02 (d, J = 8.1 Hz, 1H), 5.37 (qd, J = 13.2, 11.3, 6.6 Hz, 10H), 4.60 - 4.44 (m, 2H), 4.43 - 4.31 (m, 1H), 3.86 - 3.67 (m, 2H), 2.88 - 2.75 (m, 8H), 2.27 - 2.19 (m, 2H), 2.15 - 2.01 (m, 4H), 1.70 (q, J = 6.9, 6.4 Hz, 2H), 1.03 - 0.84 (m, 3H). HRMS (ESI): calcd for C 30 H 39 FN2O6[M+H] +543.2865, found 543.2868.

[0928] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphanyl)oxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propyl 4-fluoro-3-nitrobenzoate 5b-4

[0929]

[0930] 4b-4 was treated as described for 5a-1 to afford the title compound as a brown oil in 76% yield. 1 H NMR(400MHz,Chloroform-d)δ8.70(dd,J=7.2,2.2Hz,1H),8.31(ddd,J=8.8,4.3,2.2Hz,1H),7.38(dd,J=10.2,8 .7Hz,1H),6.78–6.61(m,1H),5.48(d,J=7.5Hz,1H),5.43–5.30(m,10H),4.57(s,1H),4.45(dd,J=6.4,2.6Hz,2H ),4.35(dtd,J=23.3,8.3,3.4Hz,2H),4.27–4.07(m,3H),2.80(dq,J=18.4,6.4,5.9Hz,8H),2.29–2.17(m,2H),2 .13–2.02(m,4H),1.71(tq,J=17.2,9.2,8.3Hz,2H),1.53–1.41(m,27H),1.04–0.93(m,3H).HRMS(ESI):calcdfor C 46 H 69 FN3O 12 P[M+Na] + 928.4495, found 928.4499.

[0931] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3- oxopropoxy)phosphoryl)oxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17- pentaenamido)propyl 4-fluoro-3-nitrobenzoate 6b-4

[0932]

[0933] Treatment of 5b-4 according to the procedure described for 6a-1 gave the target compound as a brown oil in 21% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.70 (dd, J = 7.2, 2.2 Hz, 1H), 8.31 (ddd, J = 8.8, 4.3, 2.2 Hz, 1H), 7.43 - 7.34 (m, 1H), 6.78 - 6.61 (m, 1H), 5.54 - 5.24 (m, 11H), 4.57 (s, 1H), 4.45 (dd, J = 6.4, 2.6 Hz, 2H), 4.40 - 4.29 (m, 2H), 4.28 - 4.07 (m, 3H), 2.80 (dq, J = 18.4, 6.4, 5.9 Hz, 8H), 2.29 - 2.17 (m, 2H), 2.13 - 2.02 (m, 4H), 1.78 - 1.62 (m, 2H), 1.53 - 1.41 (m, 27H), 1.04 - 0.93 (m, 3H). HRMS (ESI): calcd for C 46 H 69 FN3O 13 P[M+Na] + 944.4444, found 944.4440.

[0934] O-(((R)-3-((4-fluoro-3-nitrobenzoyl)oxy)-2-((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenamido)propoxy)(hydroxy)phosphoryl)-D-serine B5

[0935]

[0936] Treatment of 6b-4 according to the procedure described for A11 gave the target compound as a dark brown oil in 41% yield. 1H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 8.67 - 8.57 (m, 1H), 8.53 - 8.44 (m, 3H), 8.36 (br dd, J = 2.4, 6.8 Hz, 1H), 8.24 - 8.01 (m, 1H), 7.83 - 7.63 (m, 1H), 5.65 - 4.98 (m, 5H), 4.40 (br d, J = 8.0 Hz, 1H), 4.30 (br s, 3H), 4.24 - 4.10 (m, 2H), 4.07 - 3.95 (m, 2H), 2.83 - 2.70 (m, 2H), 2.22 - 1.95 (m, 6H), 1.81 - 1.40 (m, 7H), 1.39 (br s, 6H), 0.96 - 0.82 (m, 3H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 172.92, 168.80, 163.64, 156.56, 137.52, 137.42, 131.97, 129.73, 128.55, 128.41, 128.33, 128.19, 128.08, 127.74, 127.16, 119.68, 65.10, 64.46, 63.87, 53.04, 48.07, 35.31, 26.63, 25.70, 25.59, 24.28, 20.45, 14.45. HRMS (ESI): calcd for C 33 H 45 FN3O 11 P[M+H] + 710.2849, found 710.2834.

[0937] Synthesis of compound B6 of Example 36

[0938] (R,E)-N-(1-(benzyloxy)-3-hydroxypropan-2-yl)-3-(3,4-dimethoxyphenyl)acrylamide 2b-2

[0939]

[0940] Following the procedure described for 2a-1, 3,4-dimethoxycinnamic acid and 1b were treated to give the target compound as a white solid in 90% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.56 (d, J = 15.5 Hz, 1H), 7.40 - 7.28 (m, 5H), 7.08 (dd, J = 8.3, 2.0 Hz, 1H), 7.02 (d, J = 2.0 Hz, 1H), 6.85 (d, J = 8.3 Hz, 1H), 6.30 (d, J = 15.5 Hz, 1H), 4.54 (s, 2H), 4.24 (dt, J = 8.0, 4.1 Hz, 1H), 3.91 (s, 7H), 3.79 - 3.68 (m, 3H). HRMS (ESI): calcd for C 21 H 25 NO5[M+Na] + 394.1625, found 394.1626.

[0941] (S)-3-(benzyloxy)-2-((E)-3-(3,4-dimethoxyphenyl)acrylamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 3b-5

[0942]

[0943] Following the procedure described in 3a-4, EPA and 2b-2 were treated to give the target compound as yellow oil in 87% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.56 (d, J = 15.5 Hz, 1H), 7.40 - 7.28 (m, 5H), 7.08 (dd, J = 8.3, 2.0 Hz, 1H), 7.02 (d, J = 2.0 Hz, 1H), 6.86 (d, J = 8.3 Hz, 1H), 6.25 (d, J = 15.5 Hz, 1H), 5.44 - 5.26 (m, 9H), 4.53 (d, J = 1.6 Hz, 2H), 4.27 (ddd, J = 59.7, 11.1, 6.2 Hz, 2H), 3.91 (d, J = 1.3 Hz, 6H), 3.70 - 3.52 (m, 2H), 2.86 - 2.75 (m, 8H), 2.33 - 2.27 (m, 2H), 2.13 - 2.02 (m, 4H), 1.74 - 1.63 (m, 2H), 0.97 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 41 H 53 NO5[M+Na] + 678.3765, found 678.3766.

[0944] (S)-2-((E)-3-(3,4-dimethoxyphenyl)acrylamido)-3-hydroxypropyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 4b-5

[0945]

[0946] Following the procedure described in 4b-1, 3b-5 was treated to give the target compound as a yellow oil in 66% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.56 (d, J = 15.5 Hz, 1H), 7.06 (dd, J = 8.3, 2.0 Hz, 1H), 6.99 (d, J = 1.9 Hz, 1H), 6.81 (d, J = 8.3 Hz, 1H), 6.35 (d, J = 7.5 Hz, 1H), 6.30 (d, J = 15.5 Hz, 1H), 5.51 - 5.20 (m, 10H), 4.37 - 4.24 (m, 3H), 3.88 (d, J = 4.3 Hz, 6H), 3.72 (qd, J = 11.6, 3.9 Hz, 2H), 2.89 - 2.73 (m, 8H), 2.36 (t, J = 7.6 Hz, 2H), 2.14 - 2.01 (m, 4H), 1.70 (p, J = 7.5 Hz, 2H), 0.96 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 34 H 47 NO6[M+Na] + 588.3296, found 588.3291.

[0947] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-((E)-3-(3,4-dimethoxyphenyl)acrylamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 5b-5

[0948]

[0949] Following the procedure described in 5a-1, 4b-5 was treated to give the target compound as a yellow oil in 59% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.57 (d, J = 15.6 Hz, 1H), 7.08 (dd, J = 8.3, 2.0 Hz, 1H), 7.04 - 7.02 (m, 1H), 6.84 (d, J = 8.3 Hz, 1H), 6.63 (dd, J = 32.3, 8.1 Hz, 1H), 6.32 (dd, J = 15.5, 2.4 Hz, 1H), 5.54 (t, J = 7.6 Hz, 1H), 5.44 - 5.24 (m, 10H), 4.52 (s, 1H), 4.45 - 4.00 (m, 6H), 3.90 (d, J = 1.6 Hz, 6H), 2.86 - 2.74 (m, 8H), 2.34 (t, J = 7.6 Hz, 2H), 2.15 - 2.00 (m, 4H), 1.69 (p, J = 7.6 Hz, 2H), 1.46 (td, J = 9.8, 6.1 Hz, 27H), 0.96 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 50 H 77 N2O 12 P[M+Na] + 951.5106, found 951.5094.

[0950] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-((E)-3-(3,4-dimethoxyphenyl)acrylamido)propyl(5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoate 6b-5

[0951]

[0952] According to the method described in 6a-1, 5b-5 was treated to give the target compound, yellow oil, yield 54%. 1H NMR (400 MHz, Chloroform-d) δ 7.57 (d, J = 15.6 Hz, 1H), 7.08 (dd, J = 8.3, 2.0 Hz, 1H), 7.04 - 7.02 (m, 1H), 6.84 (d, J = 8.3 Hz, 1H), 6.63 (dd, J = 32.3, 8.1 Hz, 1H), 6.32 (dd, J = 15.5, 2.4 Hz, 1H), 5.54 (t, J = 7.6 Hz, 1H), 5.44 - 5.23 (m, 10H), 4.52 (s, 1H), 4.45 - 4.00 (m, 6H), 3.90 (d, J = 1.6 Hz, 6H), 2.88 - 2.72 (m, 8H), 2.34 (t, J = 7.6 Hz, 2H), 2.15 - 2.00 (m, 4H), 1.69 (p, J = 7.6 Hz, 2H), 1.46 (td, J = 9.8, 9.4, 6.1 Hz, 27H), 0.96 (t, J = 7.5 Hz, 3H). HRMS (ESI): calcd for C 50 H 77 N2O 13 P[M+H] + 945.5236, found 945.5244.

[0953] O-(((R)-2-((E)-3-(3,4-dimethoxyphenyl)acrylamido)-3-(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-D-serine B6

[0954]

[0955] Following the procedure described in A11, 6b-5 was treated to give the target compound as a yellow solid in 22% yield. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.69 (br s, 2H), 7.60 (br d, J = 15.6 Hz, 1H), 7.48 - 7.33 (m, 1H), 7.14 (br d, J = 8.4 Hz, 1H), 7.05 (br s, 1H), 6.91 (br d, J = 8.4 Hz, 1H), 6.42 (br d, J = 16.0 Hz, 1H), 5.57 - 5.03 (m, 5H), 4.72 - 4.42 (m, 4H), 4.41 (br s, 4H), 3.92 (br d, J = 10.4 Hz, 6H), 2.91 - 2.51 (m, 4H), 2.49 - 2.19 (m, 3H), 2.18 - 1.89 (m, 4H), 1.86 (br d, J = 4.4 Hz, 6H), 1.40 - 1.14 (m, 4H), 1.00 - 0.83 (m, 3H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.90, 170.16, 168.62, 151.47, 148.73, 145.63, 132.10, 129.42, 128.62, 128.35, 128.07, 127.97, 127.90, 126.73, 123.96, 114.60, 111.39, 65.58, 65.25, 63.58, 63.09, 55.94, 55.78, 54.30, 33.54, 29.72, 26.25, 25.53, 25.49, 24.46, 20.51, 14.12. HRMS (ESI): calcd for C 37 H 53 N2O 11 P[M+H] + 733.3460, found 733.3452.

[0956] Synthesis of compound B7 of Example 37

[0957] ethyl 4-((tert-butoxycarbonyl)(4-dodecylpiperazin-1-yl)amino)benzoate1a-24

[0958]

[0959] tert-Butyl N-(4-dodecylpiperazin-1-yl)carbamate (3.00 g, 8.12 mmol, 1.00 eq.), ethyl 4-iodobenzoate (3.36 g, 12.0 mmol, 2.03 mL, 1.50 eq.), cuprous iodide (1.55 g, 8.12 mmol, 1.00 eq.), cesium carbonate (5.29 g, 16.2 mmol, 2.00 eq.) and 1,10-phenanthroline (2.93 g, 16.2 mmol, 2.00 eq.) were dissolved in N,N-dimethylformamide (50.0 mL) and stirred at 100 °C under nitrogen for 12 h. The reaction mixture was filtered, the filtrate was diluted with water (50.0 mL), extracted with ethyl acetate (50.0 mL x 3), the combined organic layers were washed with brine (50.0 mL x 3), dried over sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel chromatography (PE / EA = 5 / 1 ~ 3 / 1) to give ethyl 4-[tert-butoxycarbonyl-(4-dodecylpiperazin-1-yl)amino]benzoate (2.06 g, 3.98 mmol, 49% yield) as a yellow oil. 1 H NMR (400 MHz, CDCl3) δ = 8.03-7.95 (m, 2H), 7.51 (d, J = 8.8 Hz, 2H), 4.37 (q, J = 7.2 Hz, 2H), 3.18 (br s, 4H), 2.54 (br s, 4H), 2.39-2.27 (m, 2H), 1.69 (br s, 1H), 1.54 (s, 9H), 1.46 (br d, J = 2.8 Hz, 2H), 1.39 (t, J = 7.2 Hz, 3H), 1.27 (s, 18H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 30 H 51 N3O4[M+H] + 518.3952, found 518.3948.

[0960] 4-((tert-butoxycarbonyl)(4-dodecylpiperazin-1-yl)amino)benzoic acid1a-25

[0961]

[0962] To a solution of 1a-24 (2.00 g, 3.86 mmol, 1.00 eq.) in methanol (5.00 mL) and tetrahydrofuran (10.0 mL) was added lithium hydroxide (2.00 M, 11.6 mL, 6.00 eq.) at 0 °C. Then, the mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched by the addition of hydrochloric acid solution (1.00 M, 2.00 mL) at 0 °C, then extracted with ethyl acetate (10.0 mL x 3). The combined organic layers were washed with brine (10.0 mL x 3), dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The crude product was used in the next step without further purification to give compound 4-[tert-butoxycarbonyl-(4-dodecylpiperazin-1-yl)amino]benzoic acid (1.85 g, 3.68 mmol, 95% yield) as a yellow solid. 1 H NMR (400 MHz, CDC13) δ = 12.65 - 11.89 (m, 1H), 8.01 (d, J = 8.8 Hz, 2H), 7.47 (br d, J = 8.8 Hz, 2H), 4.06 - 3.85 (m, 1H), 3.72 - 3.50 (m, 2H), 3.48 - 3.21 (m, 2H), 3.13 - 2.90 (m, 3H), 1.97 - 1.80 (m, 2H), 1.58 (s, 9H), 1.37 - 1.19 (m, 18H), 0.87 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 28 H 47 N3O4[M+H] + 490.3639, found 490.3640.

[0963] (R)-N-(1-((tert-butyldiphenylsilyl)oxy)-3-hydroxypropan-2-yl)stearamide 2b-3

[0964]

[0965] Stearic acid and (R)-2-amino-3-((tert-butyldiphenylsilyl)oxy)propan-1-ol were treated according to the method described in 2a-1 to give the target compound as a colorless transparent oil in 79% yield. 1H NMR (400 MHz, CDC13) δ = 7.64 (dd, J = 1.2, 8.0 Hz, 4H), 7.50 - 7.38 (m, 6H), 6.05 (br d, J = 7.2 Hz, 1H), 4.06 - 3.97 (m, 1H), 3.89 - 3.75 (m, 3H), 3.67 (dd, J = 4.4, 11.2 Hz, 1H), 3.01 - 2.73 (m, 1H), 2.21 - 2.11 (m, 2H), 1.64 - 1.56 (m, 2H), 1.33 - 1.24 (m, 28H), 1.09 (s, 9H), 0.89 (t, J = 6.8 Hz, 3H). HRMS (ESI): calcd for C 37 H 61 NO3Si[M + Na] + 618.4313, found 618.4310.

[0966] (R)-3-((tert-butyldiphenylsilyl)oxy)-2-stearamidopropyl 4-((tert-butoxycarbonyl)(4-dodecylpiperazin-1-yl)amino)benzoate 3b-6

[0967]

[0968] According to the method described in 3a-4, 1a-25 and 2b-3 were treated to obtain the target compound, light yellow oil, yield 65%. 1 H NMR (400 MHz, CDC13) δ = 7.89 (d, J = 8.8 Hz, 2H), 7.66 - 7.60 (m, 4H), 7.52 (d, J = 8.8 Hz, 2H), 7.44 - 7.32 (m, 6H), 5.82 (br d, J = 8.0 Hz, 1H), 4.56 - 4.33 (m, 3H), 3.93 - 3.71 (m, 2H), 3.20 (br s, 4H), 2.55 (br s, 4H), 2.39 - 2.29 (m, 2H), 2.13 (t, J = 7.6 Hz, 2H), 1.55 (s, 9H), 1.47 (br s, 2H), 1.33 - 1.22 (m, 48H), 1.08 (s, 9H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 65 H 106 N4O6Si[M + Na] + 1089.7774, found 1089.7764.

[0969] (S)-3-hydroxy-2-stearamidopropyl 4-((tert-butoxycarbonyl)(4-dodecylpiperazin-1-yl)amino)benzoate 4b-6

[0970]

[0971] Compound 4b-6 was prepared according to the procedure described in Example 1. Compound 3b-6 (2.00 g, 1.87 mmol, 1.00 eq.) was dissolved in tetrahydrofuran (20.0 mL), and tetrabutylammonium fluoride (1.00 M, 3.75 mL, 2.00 eq.) was added at 0 °C. Then, the mixture was stirred at 25 °C for 1 h. The reaction mixture was diluted with water (50.0 mL) and extracted with ethyl acetate (30.0 mL x 3). The combined organic layers were washed with brine (50.0 mL x 3), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (PE / EA = 3 / 1 ~ 2 / 1) to give compound 4b-6 as a white solid in 87% yield. 1 H NMR (400 MHz, CDC13) δ = 7.96 (d, J = 8.8 Hz, 2H), 7.57 (d, J = 8.8 Hz, 2H), 6.05 (br d, J = 7.6 Hz, 1H), 4.54-4.37 (m, 2H), 4.35-4.26 (m, 1H), 3.80-3.63 (m, 2H), 3.40-2.99 (m, 4H), 2.54 (br s, 4H), 2.38-2.31 (m, 2H), 2.22 (t, J = 7.6 Hz, 2H), 1.68-1.61 (m, 3H), 1.56 (s, 9H), 1.47 (br s, 2H), 1.36-1.20 (m, 46H), 0.89 (t, J = 6.4 Hz, 6H). HRMS (ESI): calcd for C 49 H 88 N4O6[M+Na] + 851.6596, found 851.6599.

[0972] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphaneyl)oxy)-2-stearamidopropyl 4-((tert-butoxycarbonyl)(4-dodecylpiperazin-1-yl)amino)benzoate 5b-6

[0973]

[0974] Treatment of 4b-6 according to the method described in 5a-1 gave the target compound as a colorless transparent oil in 36% yield. 1 HNMR (400 MHz, CDC13) δ = 7.95 (d, J = 8.8 Hz, 2H), 7.53 (d, J = 8.8 Hz, 2H), 6.22-6.02 (m, 1H), 5.64-5.37 (m, 1H), 4.57-4.22 (m, 4H), 4.05 (br d, J = 2.8 Hz, 2H), 3.96-3.79 (m, 2H), 3.18 (br s, 4H), 2.54 (br s, 4H), 2.39-2.29 (m, 2H), 2.27-2.16 (m, 2H), 1.65-1.60 (m, 4H), 1.54 (s, 9H), 1.48-1.45 (m, 18H), 1.39 (s, 9H), 1.31-1.23 (m, 46H), 0.89 (t, J = 6.8 Hz, 6H). HRMS (ESI): calcd for C 65 H 118 N5O 12 P[M+Na] + 1214.8407, found 1214.8411.

[0975] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-3-oxopropoxy)phosphoryl)oxy)-2-stearamidopropyl 4-((tert-butoxycarbonyl)(4-dodecyl-piperazin-1-yl)amino)benzoate 6b-6

[0976]

[0977] Treatment of 5b-6 according to the method described in 6a-1 gave the target compound as a colorless transparent oil in 43% yield. 1HNMR (400 MHz, CDC13) δ = 7.94 (d, J = 8.2 Hz, 2H), 7.54 (br d, J = 8.1 Hz, 2H), 6.56 (br s, 1H), 5.63-5.43 (m, 1H), 4.55 (br d, J = 3.7 Hz, 1H), 4.48-4.18 (m, 6H), 4.17-4.12 (m, 1H), 3.19 (br s, 4H), 2.54 (br d, J = 0.9 Hz, 4H), 2.40-2.30 (m, 2H), 2.28-2.15 (m, 2H), 1.61 (br d, J = 7.2 Hz, 4H), 1.54 (s, 9H), 1.51-1.44 (m, 27H), 1.33-1.21 (m, 46H), 0.88 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 65 H 118 N5O 13 P[M+Na] + 1230.8356, found 1230.8362.

[0978] O-(((R)-3-((4-((4-dodecylpiperazin-1-yl)amino)benzoyl)oxy)-2-stearamidopropoxy)(hydroxy)phosphoryl)-D-serine B7

[0979]

[0980] According to the method described in A1, 6b-6 was treated to obtain the target compound, yellow solid, yield 75%. 1 H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 8.89-8.61 (m, 1H), 7.85 (br s, 2H), 7.72 (br d, J = 2.0 Hz, 1H), 7.59 (br s, 2H), 7.49-7.36 (m, 1H), 6.91 (br d, J = 8.0 Hz, 2H), 4.67-4.30 (m, 6H), 4.21 (br d, J = 4.4 Hz, 2H), 3.78 (br d, J = 10.4 Hz, 2H), 3.44 (br d, J = 10.4 Hz, 2H), 3.34-3.09 (m, 6H), 2.36 (br s, 2H), 1.74 (br s, 2H), 1.57 (br s, 2H), 1.26 (br s, 46H), 0.92-0.84 (m, 6H). 13C NMR (101 MHz, CDC13and 20 μL TFA) δ = 179.01, 168.42, 164.69, 149.67, 132.13, 132.09, 120.08, 113.74, 87.82, 65.20, 62.96, 58.00, 54.24, 52.29, 52.24, 51.41, 49.56, 35.58, 31.89, 31.84, 29.67, 29.50, 29.47, 29.31, 29.25, 29.13, 27.31, 25.85, 22.62, 13.95. HRMS (ESI): calcd for C 47 H 86 N5O9P[M+H] + 896.6236, found 896.6221.

[0981] Synthesis of compound C1 of example 38

[0982] tert-butyl (4-nitrobenzoyl)-D-serinate Id-1

[0983]

[0984] (R)-2-amino-3-hydroxypropionic acid tert-butyl ester hydrochloride (0.50 g, 2.53 mmol, 1.10 eq.) was dissolved in tetrahydrofuran (20 mL) solution, the solution PH was adjusted to 10-11 by adding saturated sodium bicarbonate solution (10 mL) and stirring for 5 min. 4-nitrobenzoyl chloride (0.43 g, 2.30 mmol, 1.00 eq.) was added under ice bath, stirring at room temperature for 2 h, monitoring the reaction by TLC, after the reaction was completed, extracted with dichloromethane (30 mL x 3), combined organic layer, washed with saturated brine (30 mL x 3), then dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain a residue. The residue was purified by silica gel column (PE / EA = 2 / 1-1 / 2) to obtain compound Id-1, white solid, yield 86%. 1 H NMR (400 MHz, Chloroform-d) δ 8.33 - 8.27 (m, 2H), 8.03 - 7.97 (m, 2H), 7.19 (d, J = 6.9 Hz, 1H), 4.74 (dt, J = 6.9, 3.5 Hz, 1H), 4.07 (t, J = 4.3 Hz, 2H), 2.41 (d, J = 4.7 Hz, 1H), 1.52 (s, 9H). HRMS (ESI): calcd for C 14 H 18 N2O6[M+Na] +333.1057, found 333.1056.

[0985] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(4-nitrobenzamido)-3-oxopropoxy)

[0986] phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-1

[0987]

[0988] Dissolve 1-tert-butoxy-N,N,N',N'-tetraisopropylphosphorodiamidite (660 mg, 2.16 mmol, 1.00 eq.) in dichloromethane (10.0 mL) and toluene (1.00 mL). To this solution, add 1d-1 (470 mg, 1.51 mmol, 0.70 eq.). To eliminate trace amount of water, evaporate the resulting solution under vacuum. Dissolve the residue in anhydrous dichloromethane (10.0 mL) under nitrogen protection, add 1H-tetrazole (182 mg, 2.60 mmol, 1.20 eq.) at room temperature, after a few minutes, a white solid precipitate appears. Stir the whole under nitrogen at room temperature for 3 h. Add 4a-10 (1.07 g, 1.62 mmol, 0.75 eq.) and 1H-tetrazole (303 mg, 4.33 mmol, 2.00 eq.) dissolved in dichloromethane (3.00 mL) under nitrogen. Then, stir the mixture at room temperature overnight. Check the reaction completion with TLC plate, after the reaction is finished, quench the reaction mixture with sodium bicarbonate solution (30.0 mL), then extract with dichloromethane (30.0 mL x 3). Wash the combined organic layers with brine (30.0 mL x 2), dry over sodium sulfate, concentrate under reduced pressure to get a residue. Purify the residue by silica gel column chromatography (PE / EA = 20 / 1 ~ 5 / 1, 0.1% TEA) to get compound 2d-1 as colorless transparent oil with a yield of 54%. 1H NMR (400 MHz, Chloroform-d) δ 8.33 - 8.27 (m, 2H), 8.20 (d, J = 7.4 Hz, 1H), 8.17 - 8.10 (m, 2H), 5.44 - 5.27 (m, 20H), 5.22 (p, J = 5.1 Hz, 1H), 4.85 (ddt, J = 7.6, 3.8, 2.1 Hz, 1H), 4.50 (td, J = 11.6, 3.6 Hz, 1H), 4.40 (ddd, J = 11.1, 7.4, 2.3 Hz, 1H), 4.31 (dd, J = 12.0, 4.2 Hz, 1H), 4.13 (ddd, J = 10.4, 5.7, 2.4 Hz, 3H), 2.88 - 2.75 (m, 16H), 2.38 - 2.27 (m, 4H), 2.14 - 2.00 (m, 8H), 1.69 (dt, J = 15.3, 7.7 Hz, 4H), 1.51 - 1.46 (m, 18H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 89 N2O 12 P[M+Na] + 1095.6045, found 1095.6045.

[0989] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(4-nitrobenzamido)-3-oxopropoxy)

[0990] phosphoryl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)3d-1

[0991]

[0992] Following the procedure described in 6a-1, 2d-1 was treated to give the target compound as a colorless transparent oil in 34% yield. 1HNMR(400MHz,Chloroform-d)δ8.33–8.27(m,2H),8.20(d,J=7.4Hz,1H),8.17–8.10(m,2H),5.46–5.25(m,20H ),5.22(p,J=5.1Hz,1H),4.85(ddt,J=7.6,3.8,2.1Hz,1H),4.50(td,J=11.6,3.6Hz,1H),4.40(ddd,J=11.1,7 .4,2.3Hz,1H),4.31(dd,J=12.0,4.2Hz,1H),4.13(ddd,J=10.4,5.7,2.4Hz,3H),2.88–2.75(m,16H),2.38–2. 27(m,4H),2.14–2.00(m,8H),1.74–1.62(m,4H),1.54–1.43(m,18H),0.97(t,J=7.5Hz,6H).HRMS(ESI):calcd forC 61 H 89 N2O 13 P[M+Na] + 1111.5994,found 1111.5995.

[0993] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(4-nitrobenzoyl)-D-serine C1

[0994]

[0995] 3d-1 was treated according to the method described in A11 to obtain the target compound as a dark brown oil in a yield of 41%. 1 H NMR (400MHz, DMSO-d6 and 20μL TFA)δ=9.36-8.79(m,1H),8.36-8.28(m,2H),8.20-7.96(m,2H),5.68-5.02(m,10H),4.79-4.66(m,1H),4.40-4.15(m,3H),4.14- 3.89(m,3H),3.87-3.74(m,1H),2.91-2.65(m,5H),2.42-2.12(m,8H),2.12-1.80(m,9H),1.77-1.01(m,22H),0.94-0.79(m,6H). 13C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 174.79, 172.03, 170.66, 165.40, 149.58, 140.09, 131.98, 129.59, 129.42, 128.55, 128.42, 128.28, 128.12, 127.36, 124.02, 123.94, 62.87, 62.67, 61.45, 56.34, 53.93, 33.47, 26.50, 26.38, 25.62, 24.87, 24.78, 20.47, 14.49. HRMS (ESI): calcd for C 53 H 73 N2O6 13 P[M-H] - 975.4778, found 975.4745.

[0996] Synthesis of compound C2 of example 39

[0997] tert-butyl (3-nitrobenzoyl)-D-serinate Id-2

[0998]

[0999] The procedure described for Id-1 was followed to treat 3-nitrobenzoyl chloride to give the target compound as a white solid in 98% yield. 1 H NMR (400 MHz, Chloroform-d) δ 8.68 (t, J = 2.0 Hz, 1H), 8.38 (ddd, J = 8.3, 2.2, 1.1 Hz, 1H), 8.18 (dt, J = 8.0, 1.4 Hz, 1H), 7.66 (t, J = 8.0 Hz, 1H), 7.25 (s, 1H), 4.77 (dt, J = 7.1, 3.6 Hz, 1H), 4.08 (s, 2H), 1.53 (s, 9H). HRMS (ESI): calcd for C 14 H 18 N2O6 [M+Na] + 333.1057, found 333.1052.

[1000] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(3-nitrobenzamido)-3-oxopropoxy)phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-2

[1001]

[1002] Following the procedure described in 2d-1, 1d-2 was treated to give the target compound as a colorless transparent oil in 23% yield. 1 HNMR (400 MHz, Chloroform-d) δ 8.85 - 8.81 (m, 1H), 8.36 (ddd, J = 8.2, 2.3, 1.0 Hz, 1H), 8.31 - 8.26 (m, 1H), 8.22 (d, J = 7.7 Hz, 1H), 7.68 - 7.62 (m, 1H), 5.44 - 5.27 (m, 20H), 5.23 (d, J = 5.7 Hz, 1H), 4.85 (dd, J = 12.2, 7.2 Hz, 1H), 4.62 - 4.46 (m, 2H), 4.45 - 4.36 (m, 1H), 4.31 (dd, J = 12.0, 4.1 Hz, 1H), 4.20 - 4.05 (m, 2H), 2.88 - 2.73 (m, 16H), 2.31 (td, J = 7.9, 3.3 Hz, 4H), 2.06 (ddd, J = 10.8, 7.9, 5.9 Hz, 8H), 1.73 - 1.60 (m, 4H), 1.51 (d, J = 2.3 Hz, 9H), 1.49 (dd, J = 3.8, 0.6 Hz, 9H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 89 N2O 12 P[M+Na] + 1095.6045, found 1095.6044.

[1003] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(3-nitrobenzamido)-3-oxopropoxy)

[1004] (5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icos-5,8,11,14,17-pentaenoate) 3d-2

[1005]

[1006] Following the procedure described in 6a-1, 2d-2 was treated to give the target compound as a colorless transparent oil in 47% yield. 1 HNMR (400 MHz, Chloroform-d) δ 8.85 - 8.81 (m, 1H), 8.36 (ddd, J = 8.2, 2.3, 1.0 Hz, 1H), 8.31 - 8.26 (m, 1H), 8.22 (d, J = 7.7 Hz, 1H), 7.68 - 7.62 (m, 1H), 5.45 - 5.26 (m, 20H), 5.26 - 5.19 (m, 1H), 4.84 (t, J = 9.7 Hz, 1H), 4.58 - 4.46 (m, 2H), 4.45 - 4.37 (m, 1H), 4.31 (dd, J = 12.0, 4.1 Hz, 1H), 4.20 - 4.05 (m, 2H), 2.86 - 2.76 (m, 16H), 2.31 (td, J = 7.9, 3.3 Hz, 4H), 2.06 (ddd, J = 10.8, 7.9, 5.9 Hz, 8H), 1.73 - 1.60 (m, 4H), 1.52 - 1.48 (m, 18H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 89 N2O 13 P[M+Na] + 1111.5994, found 1111.5999.

[1007] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(3-nitrobenzoyl)-D-serine C2

[1008]

[1009] Following the procedure described in A11, 3d-2 was treated to give the target compound as a dark brown oil in 77% yield. 1H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 9.34 (br d, J = 7.6 Hz, 1H), 8.76 - 8.72 (m, 1H), 8.40 (td, J = 1.2, 8.0 Hz, 1H), 8.33 (br d, J = 7.6 Hz, 1H), 7.79 (br t, J = 8.0 Hz, 1H), 5.67 - 5.03 (m, 20H), 4.75 (br d, J = 4.8 Hz, 1H), 4.36 - 4.18 (m, 3H), 4.11 - 3.92 (m, 3H), 3.82 (br d, J = 5.2 Hz, 1H), 2.85 - 2.72 (m, 12H), 2.34 - 2.23 (m, 4H), 2.07 - 1.96 (m, 8H), 1.61 - 1.48 (m, 6H), 1.29 - 1.21 (m, 2H), 0.95 - 0.86 (m, 6H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 174.77, 172.07, 170.70, 164.75, 148.24, 134.41, 134.26, 131.96, 128.67, 128.53, 128.42, 128.30, 128.27, 128.11, 127.35, 122.63, 69.93, 65.58, 61.45, 56.38, 53.94, 33.46, 33.30, 33.13, 26.50, 26.36, 25.60, 24.86, 24.75, 24.70, 20.45, 14.45. HRMS (ESI): calcd for C 53 H 73 N2O 13 P[M-H] - 975.4778, found 975.4763.

[1010] Synthesis of compound C3 of example 40

[1011] tert-butyl (3,5-dinitrobenzoyl)-D-serinate Id-3

[1012]

[1013] The procedure described for Id-1 was followed to treat 3,5-dinitrobenzoyl chloride to give the target compound as a white solid in 56% yield. 1H NMR (400 MHz, Chloroform-d) δ 9.19 (t, J = 2.1 Hz, 1H), 9.02 (d, J = 2.1 Hz, 2H), 7.44 (d, J = 7.2 Hz, 1H), 4.81 (dt, J = 7.0, 3.3 Hz, 1H), 4.18 - 4.04 (m, 2H), 2.33 (s, 1H), 1.53 (s, 9H). HRMS (ESI): calcd for C 14 H 17 N3O8[M+Na] + 378.0908, found 378.0900.

[1014] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(3,5-dinitrobenzamido)-3-oxopropoxy)

[1015] phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-3

[1016]

[1017] According to the method described in 2d-1, 1d-3 was treated to obtain the target compound, colorless transparent oil, colorless transparent oil, yield 14%. 1 H NMR (400 MHz, Chloroform-d) δ 9.20 (d, J = 2.1 Hz, 2H), 9.14 (t, J = 2.1 Hz, 1H), 8.81 (d, J = 7.4 Hz, 1H), 5.42 - 5.19 (m, 21H), 4.85 (ddt, J = 7.4, 3.7, 2.0 Hz, 1H), 4.54 - 4.36 (m, 2H), 4.28 (ddd, J = 12.0, 4.2, 2.0 Hz, 1H), 4.13 (dtd, J = 14.5, 11.3, 6.3 Hz, 3H), 2.85 - 2.73 (m, 16H), 2.36 - 2.25 (m, 4H), 2.11 - 2.01 (m, 8H), 1.73 - 1.59 (m, 4H), 1.49 (d, J = 3.0 Hz, 18H), 0.95 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 88 N3O 14 P[M+Na]+ 1140.5896, found 1140.5894.

[1018] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(3,5-dinitrobenzamido)-3-oxopropoxy)

[1019] phosphoryl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)3d-3

[1020]

[1021] According to the method described in 6a-1, 2d-3 was treated to obtain the target compound, colorless transparent oil, yield 72%. 1 HNMR (400 MHz, Chloroform-d) δ 9.22 (d, J = 2.1 Hz, 2H), 9.16 (t, J = 2.1 Hz, 1H), 8.82 (d, J = 7.4 Hz, 1H), 5.43 - 5.28 (m, 20H), 5.28 - 5.21 (m, 1H), 4.86 (ddt, J = 7.4, 3.7, 2.0 Hz, 1H), 4.56 - 4.47 (m, 1H), 4.42 (ddd, J = 11.2, 7.4, 2.1 Hz, 1H), 4.30 (ddd, J = 12.0, 4.2, 2.0 Hz, 1H), 4.15 (dtd, J = 14.5, 11.3, 6.3 Hz, 3H), 2.86 - 2.76 (m, 16H), 2.38 - 2.26 (m, 4H), 2.13 - 2.02 (m, 8H), 1.67 (dt, J = 14.7, 7.4 Hz, 4H), 1.51 (d, J = 3.0 Hz, 18H), 0.96 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 88 N3O 15 P[M+Na] + 1156.5845, found 1156.5839.

[1022] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(3,5-dinitrobenzoyl)-D-serine C3

[1023]

[1024] The title compound was prepared according to the procedure described for Al l from 3d-3, yield 73% as dark brown oil. 1 H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 9.78 - 9.28 (m, 1H), 9.11 (br s, 2H), 8.98 (br s, 1H), 5.32 (br s, 10H), 4.80 (br d, J = 0.8 Hz, 1H), 4.30 (br d, J = 5.2 Hz, 3H), 4.01 (br s, 3H), 3.84 (br d, J = 5.2 Hz, 1H), 2.89 - 2.64 (m, 6H), 2.41 - 2.10 (m, 8H), 2.03 (br s, 4H), 1.87 - 0.99 (m, 24H), 0.87 (br dd, J = 6.4 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 171.83, 170.47, 163.10, 162.93, 148.67, 148.60, 137.09, 136.75, 131.90, 128.79, 128.48, 128.27, 128.03, 127.27, 121.44, 121.32, 67.56, 65.41, 61.30, 56.62, 54.13, 33.54, 31.65, 29.47, 26.35, 25.54, 25.45, 24.81, 20.39, 14.36. HRMS (ESI): calcd for C 53 H 72 N3O 15 P[M-H] - 1020.4628, found 1020.4610.

[1025] Synthesis of compound C4 of Example 41

[1026] tert-butyl(4-methylbenzoyl)-D-serinate 1d-4

[1027]

[1028] The procedure described in 1d-1 was followed using 4-methylbenzoyl chloride to give the title compound as a clear oil in 97% yield. 1 H NMR (400 MHz, Chloroform-d) δ 7.75 - 7.70 (m, 2H), 7.25 - 7.20 (m, 2H), 7.12 (d, J = 6.7 Hz, 1H), 4.73 (ddd, J = 6.6, 4.4, 3.2 Hz, 1H), 4.06 (dd, J = 11.2, 3.1 Hz, 1H), 3.99 (dd, J = 11.1, 4.4 Hz, 1H), 2.95 (s, 1H), 2.40 (s, 3H), 1.51 (s, 9H). HRMS (ESI): calcd for C 15 H 21 NO4[M+Na] + 302.1363, found 302.1368.

[1029] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(4-methylbenzamido)-3-oxopropoxy)

[1030] phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-4

[1031]

[1032] The procedure described in 2d-1 was followed using 1d-4 to give the title compound as a colorless clear oil in 11% yield. 1HNMR (400 MHz, Chloroform-d) δ 7.79 (dt, J = 8.4, 2.0 Hz, 2H), 7.24 (d, J = 8.0 Hz, 2H), 5.46 - 5.25 (m, 20H), 5.19 (h, J = 5.2 Hz, 1H), 4.85 (ddq, J = 10.5, 7.8, 2.8 Hz, 1H), 4.48 (ddt, J = 10.5, 9.1, 3.3 Hz, 1H), 4.37 (ddd, J = 10.6, 6.3, 2.5 Hz, 1H), 4.29 (ddt, J = 10.7, 8.3, 4.0 Hz, 1H), 4.17 - 4.01 (m, 3H), 2.87 - 2.76 (m, 16H), 2.42 - 2.26 (m, 7H), 2.13 - 2.02 (m, 8H), 1.69 (qd, J = 7.4, 5.1 Hz, 4H), 1.52 - 1.41 (m, 18H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 62 H 92 NO 10 P[M+Na] + 1064.6351, found 1064.6357.

[1033] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(4-methylbenzamido)-3-oxopropoxy)

[1034] phosphoryl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)3d-4

[1035]

[1036] According to the method described in 6a-1, 2d-4 was treated to obtain the target compound, colorless transparent oil, yield 64%. 1HNMR (400 MHz, Chloroform-d) δ 7.79 (dt, J = 8.4, 2.0 Hz, 2H), 7.24 (d, J = 8.0 Hz, 2H), 5.46 - 5.25 (m, 20H), 5.19 (h, J = 5.2 Hz, 1H), 4.85 (ddq, J = 10.5, 7.8, 2.8 Hz, 1H), 4.48 (ddt, J = 10.5, 9.1, 3.3 Hz, 1H), 4.37 (ddd, J = 10.6, 6.3, 2.5 Hz, 1H), 4.29 (ddt, J = 10.7, 8.3, 4.0 Hz, 1H), 4.15 - 4.03 (m, 3H), 2.91 - 2.74 (m, 16H), 2.40 - 2.28 (m, 7H), 2.13 - 2.02 (m, 8H), 1.69 (qd, J = 7.4, 5.1 Hz, 4H), 1.52 - 1.41 (m, 18H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 62 H 92 NO 11 P[M+Na] + 1058.6481, found 1058.6482.

[1037] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(4-methylbenzoyl)-D-serine C4

[1038]

[1039] Following the procedure described in A11, 3d-4 was treated to give the target compound as a brown oil in 39% yield. 1H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 8.28 (d, J = 7.6 Hz, 1H), 7.79 (br d, J = 8.0 Hz, 2H), 7.28 (br d, J = 8.0 Hz, 2H), 5.33 (br d, J = 4.4 Hz, 10H), 4.48 - 4.23 (m, 2H), 4.06 - 3.95 (m, 1H), 3.86 - 3.69 (m, 3H), 3.54 - 3.28 (m, 2H), 2.87 - 2.61 (m, 4H), 2.35 (s, 3H), 2.32 - 2.12 (m, 8H), 2.08 - 1.93 (m, 6H), 1.81 - 1.35 (m, 16H), 1.30 - 1.22 (m, 8H), 0.93 - 0.82 (m, 6H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 174.78, 172.46, 171.25, 166.70, 141.74, 131.97, 131.63, 129.29, 129.22, 128.77, 128.42, 128.28, 127.85, 127.80, 67.67, 65.30, 61.69, 58.90, 56.03, 33.47, 31.52, 30.21, 26.50, 25.62, 25.53, 24.86, 21.34, 20.45, 14.47. HRMS (ESI): calcd for C 54 H 76 NO 11 P[M-H] - 944.5083, found 944.5041.

[1040] Synthesis of compound C5 of example 42

[1041] tert-butyl (4-fluorobenzoyl)-D-serinate Id-5

[1042]

[1043] The procedure described in Id-1 was followed to treat 4-fluorobenzoyl chloride to give the target compound as a colorless transparent oil in 98% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.87 - 7.80 (m, 2H), 7.14 - 7.06 (m, 3H), 4.71 (ddd, J = 7.1, 4.2, 3.2 Hz, 1H), 4.02 (qd, J = 11.1, 3.7 Hz, 2H), 2.83 (s, 1H), 1.51 (s, 9H). HRMS (ESI): calcd for C 14 H 18 FNO4[M+Na] + 306.1112, found 306.1104.

[1044] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(4-fluorobenzamido)-3-oxopropoxy)

[1045] phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-5

[1046]

[1047] According to the method described in 2d-1, 1d-5 was treated to obtain the target compound, colorless transparent oil, yield 20%. 1 H NMR (400 MHz, Chloroform-d) δ 7.94 (ddd, J = 8.9, 5.3, 1.6 Hz, 2H), 7.14 - 7.07 (m, 2H), 5.46 - 5.24 (m, 20H), 5.19 (q, J = 6.4, 5.9 Hz, 1H), 4.86 - 4.77 (m, 1H), 4.46 (td, J = 10.4, 3.4 Hz, 1H), 4.36 (ddd, J = 10.7, 6.6, 2.3 Hz, 1H), 4.30 (ddd, J = 12.0, 4.3, 1.6 Hz, 1H), 4.11 (ddt, J = 10.4, 5.6, 2.3 Hz, 3H), 2.82 (ddd, J = 16.8, 8.7, 4.0 Hz, 16H), 2.30 (dt, J = 10.9, 7.7 Hz, 4H), 2.14 - 2.00 (m, 8H), 1.67 (h, J = 7.4 Hz, 4H), 1.52 - 1.42 (m, 18H), 0.96 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H89 FNO 10 P[M+Na] + 1068.6100, found 1068.6101.

[1048] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(4-fluorobenzamido)-3-oxopropoxy)

[1049] phosphoryl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)3d-5

[1050]

[1051] According to the method described in 6a-1, 2d-5 was treated to obtain the target compound, colorless transparent oil, yield 67%. 1 HNMR (400 MHz, Chloroform-d) δ 7.94 (ddd, J = 8.9, 5.3, 1.6 Hz, 2H), 7.14 - 7.08 (m, 2H), 5.44 - 5.24 (m, 20H), 5.23 - 5.15 (m, 1H), 4.86 - 4.77 (m, 1H), 4.46 (td, J = 10.4, 3.4 Hz, 1H), 4.36 (ddd, J = 10.7, 6.6, 2.3 Hz, 1H), 4.30 (ddd, J = 12.0, 4.3, 1.6 Hz, 1H), 4.11 (ddt, J = 10.4, 5.6, 2.3 Hz, 3H), 2.85 - 2.76 (m, 16H), 2.30 (dt, J = 10.8, 7.7 Hz, 4H), 2.14 - 2.00 (m, 8H), 1.72 - 1.61 (m, 4H), 1.52 - 1.42 (m, 18H), 0.96 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 89 FNO 11 P[M+Na] + 1084.6049, found 1084.6035.

[1052] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(4-fluorobenzoyl)-D-serine C5

[1053]

[1054] The compound 3d-5 was treated according to the procedure described for Al l to give the target compound as a black oil in 14% yield. 1 H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 8.94 - 8.42 (m, 1H), 7.96 (br dd, J = 5.6, 8.4 Hz, 2H), 7.31 - 7.27 (m, 2H), 6.09 - 4.39 (m, 10H), 4.31 - 4.24 (m, 2H), 4.08 - 3.92 (m, 3H), 3.85 - 3.77 (m, 2H), 3.55 - 3.44 (m, 1H), 2.89 - 2.56 (m, 4H), 2.36 - 2.16 (m, 8H), 2.10 - 1.90 (m, 6H), 1.81 - 1.15 (m, 24H), 0.94 - 0.77 (m, 6H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 172.34, 165.84, 165.68, 163.41, 130.57, 130.48, 115.80, 115.71, 115.57, 115.49, 61.59, 59.87, 56.16, 53.80, 49.67, 31.50, 29.74, 29.24, 24.75, 19.16, 17.33, 17.24, 14.63. HRMS (ESI): calcd for C 53 H 73 FNO 11 P[M-H] - 948.4833, found 948.4793.

[1055] Synthesis of compound C6 of Example 43

[1056] tert-butyl (4-bromobenzoyl)-D-serinate 1d-6

[1057]

[1058] The title compound was obtained in 98% yield as a colorless transparent oil by the procedure described for 1d-1. 1 H NMR (400 MHz, Chloroform-d) δ 7.74 - 7.66 (m, 2H), 7.64 - 7.52 (m, 2H), 7.10 (d, J = 6.7 Hz, 1H), 4.71 (ddd, J = 6.9, 4.1, 3.1 Hz, 1H), 4.11 - 3.96 (m, 2H), 2.65 (t, J = 5.8 Hz, 1H), 1.51 (s, 9H). HRMS (ESI): calcd for C 14 H 18 BrNO4[M+Na] + 366.0311, found 366.0312.

[1059] (2R)-3-((((R)-2-(4-bromobenzamido)-3-(tert-butoxy)-3-oxopropoxy)(tert-butoxy)

[1060] phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-6

[1061]

[1062] The title compound was obtained in 51% yield as a colorless transparent oil by the procedure described for 2d-1. 1 H NMR (400 MHz, Chloroform-d) δ 7.82 - 7.78 (m, 2H), 7.60 - 7.56 (m, 2H), 5.45 - 5.25 (m, 20H), 5.20 (dt, J = 10.5, 5.1 Hz, 1H), 4.82 (tq, J = 8.5, 3.7, 3.1 Hz, 1H), 4.53 - 4.42 (m, 1H), 4.40 - 4.26 (m, 2H), 4.15 - 4.07 (m, 3H), 2.86 - 2.76 (m, 16H), 2.31 (dt, J = 10.0, 7.7 Hz, 4H), 2.14 - 2.03 (m, 8H), 1.73 - 1.63 (m, 4H), 1.51 - 1.44 (m, 18H), 0.97 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 89BrNO 10 P[M+Na] + 1128.53, found 1128.5302.

[1063] (2R)-3-((((R)-2-(4-bromobenzamido)-3-(tert-butoxy)-3-oxopropoxy)(tert-butoxy)

[1064] phosphoryl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)3d-6

[1065]

[1066] According to the method described in 6a-1, 2d-6 was treated to obtain the target compound, colorless transparent oil, yield 90%. 1 HNMR (400 MHz, Chloroform-d) δ 7.79 (dd, J = 8.7, 1.1 Hz, 2H), 7.60 - 7.53 (m, 2H), 5.44 - 5.24 (m, 20H), 5.19 (h, J = 5.2 Hz, 1H), 4.86 - 4.76 (m, 1H), 4.53 - 4.25 (m, 3H), 4.17 - 4.01 (m, 3H), 2.86 - 2.76 (m, 16H), 2.35 - 2.26 (m, 4H), 2.14 - 2.00 (m, 8H), 1.73 - 1.61 (m, 4H), 1.51 - 1.41 (m, 18H), 0.96 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 89 BrNO 11 P[M+Na] + 1144.5249, found 1144.5256.

[1067] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(4-bromobenzoyl)-D-serine C6

[1068]

[1069] The compound 3d-6 was treated according to the method described in Al l to give the target compound as a black oil in 14% yield. 1 H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 9.02 - 8.48 (m, 1H), 8.12 (br d, J = 4.0 Hz, 1H), 7.83 (br d, J = 8.4 Hz, 2H), 7.68 (dt, J = 2.8, 5.6 Hz, 2H), 5.32 (br s, 10H), 4.68 (br s, 1H), 4.27 (br d, J = 7.2 Hz, 3H), 4.15 - 3.89 (m, 3H), 3.81 (br dd, J = 4.8, 12.8 Hz, 1H), 2.84 - 2.70 (m, 4H), 2.35 - 2.14 (m, 6H), 2.12 - 1.85 (m, 8H), 1.85 - 1.18 (m, 24H), 0.94 - 0.78 (m, 6H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 172.26, 170.92, 166.04, 165.93, 133.22, 131.99, 131.86, 131.77, 129.95, 128.56, 128.32, 128.24, 128.13, 125.80, 70.70, 67.48, 65.62, 62.69, 53.85, 33.45, 26.41, 25.64, 25.54, 24.88, 24.78, 20.47, 14.48. HRMS (ESI): calcd for C 53 H 73 BrNO 11 P[M-H] - 1008.4032, 1010.4011, found 1010.3984.

[1070] Synthesis of compound C7 of example 44

[1071] tert-butyl (2-fluoro-4-nitrobenzoyl)-D-serinate 1d-7

[1072]

[1073] The compound 3d-6 was treated according to the method described in Al l to give the target compound as a black oil in 14% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.29 (dd, J = 8.6, 7.4 Hz, 1H), 8.14 (dd, J = 8.7, 2.1 Hz, 1H), 8.05 (dd, J = 11.0, 2.1 Hz, 1H), 7.72 - 7.64 (m, 1H), 4.77 (dtd, J = 6.9, 3.6, 1.7 Hz, 1H), 4.08 (d, J = 3.6 Hz, 2H), 1.53 (s, 9H). HRMS (ESI): calcd for C 14 H 17 FN2O6[M+Na] + 351.0963, found 351.0951.

[1074] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(2-fluoro-4-nitrobenzamido)-3-oxopropoxy)phosphaneyl)oxy)propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)2d-7

[1075]

[1076] Following the procedure described in 2d-1, 1d-7 was treated to give the target compound as a light yellow oil in 30% yield. 1H NMR(400MHz,Chloroform-d)δ8.23(ddd,J=8.6,7.3,2.7Hz,1H),8.11(dd,J=8.6,2.1Hz,1H),8.03(dt,J=10.7,1.9Hz,1H ),7.81(ddd,J=12.3,9.7,7.2Hz,1H),5.45–5.24(m,20H),5.18(p,J=5.3Hz,1H),4.90–4.82(m,1H),4.53–4.37(m,2H),4. 30(ddd,J=11.9,6.2,4.2Hz,1H),4.16–3.99(m,3H),2.89–2.72(m,16H),2.31(qd,J=7.6,3.0Hz,4H),2.16–2.01(m,8H), 1.66(qdd,J=11.4,5.6,3.4Hz,4H),1.51(d,J=1.1Hz,9H),1.45(d,J=2.9Hz,9H),0.96(t,J=7.5Hz,6H).HRMS(ESI):calcd for C 61 H 88 FN2O 12 P[M+Na] + 1113.5951,found1113.5952.

[1077] (2R)-3-((tert-butoxy((R)-3-(tert-butoxy)-2-(2-fluoro-4-nitrobenzamido)-3-oxopropoxy)phosphoryl)oxy )propane-1,2-diyl(5E,5'E,8E,8'E,11E,11'E,14E,14'E,17E,17'E)-bis(icosa-5,8,11,14,17-pentaenoate)3d-7

[1078]

[1079] 2d-7 was treated according to the method described for 6a-1 to obtain the title compound as a light yellow oil in 38% yield. 1H NMR (400 MHz, Chloroform-d) δ 8.23 (ddd, J = 8.6, 7.3, 2.7 Hz, 1H), 8.11 (dd, J = 8.6, 2.1 Hz, 1H), 8.03 (dt, J = 10.7, 1.9 Hz, 1H), 7.81 (ddd, J = 12.3, 9.7, 7.2 Hz, 1H), 5.45 - 5.24 (m, 20H), 5.18 (p, J = 5.3 Hz, 1H), 4.90 - 4.82 (m, 1H), 4.50 - 4.36 (m, 2H), 4.30 (ddd, J = 11.9, 6.2, 4.2 Hz, 1H), 4.16 - 4.01 (m, 3H), 2.89 - 2.72 (m, 16H), 2.31 (qd, J = 7.6, 3.0 Hz, 4H), 2.13 - 2.01 (m, 8H), 1.66 (qdd, J = 11.4, 5.6, 3.4 Hz, 4H), 1.51 (d, J = 1.1 Hz, 9H), 1.45 (d, J = 2.9 Hz, 9H), 0.96 (t, J = 7.5 Hz, 6H). HRMS (ESI): calcd for C 61 H 88 FN2O 13 P[M+Na] + 1129.5900, found 1129.5910.

[1080] O-(((R)-2,3-bis(((5E,8E,11E,14E,17E)-icosa-5,8,11,14,17-pentaenoyl)oxy)propoxy)(hydroxy)phosphoryl)-N-(2-fluoro-4-nitrobenzoyl)-D-serine C7

[1081]

[1082] Following the procedure described in A11, 3d-7 was treated to give the target compound as a brown oil in 82% yield. 1H NMR (400 MHz, DMSO-d6 and 20 μL TFA) δ = 9.01 (br s, 1H), 8.19 - 8.12 (m, 2H), 7.91 - 7.85 (m, 1H), 5.90 - 4.67 (m, 10H), 4.37 - 3.93 (m, 7H), 3.87 - 3.76 (m, 1H), 2.78 (br d, J = 14.4 Hz, 4H), 2.42 - 2.10 (m, 10H), 2.07 - 1.93 (m, 6H), 1.82 - 1.37 (m, 16H), 1.31 - 1.17 (m, 6H), 0.95 - 0.79 (m, 6H). 13 C NMR (101 MHz, DMSO-d6 and 20 μL TFA) δ = 172.61, 170.46, 162.94, 160.47, 157.94, 149.84, 149.75, 131.86, 129.95, 129.78, 128.85, 128.77, 128.54, 128.32, 128.24, 128.10, 119.97, 112.65, 112.37, 70.04, 65.61, 61.46, 55.93, 53.67, 33.47, 33.14, 31.50, 30.20, 26.36, 25.61, 24.79, 20.47, 14.46. HRMS (ESI): calcd for C 53 H 72 FN2O 13 P[M-H] - 993.4683, found 993.4680.

[1083] Synthesis of compound C8 of example 45

[1084] tert-butyl (tert-butoxycarbonyl)-L-threoninate 1d-8

[1085]

[1086] Dissolve L-threonine tert-butyl ester hydrochloride (0.50 g, 2.36 mmol, 1.00 eq.) in tetrahydrofuran (20 mL). Add saturated sodium bicarbonate solution (10 mL) to adjust the solution pH to 10-11 and stir for 5 min. Add BOC anhydride (0.62 g, 2.83 mmol, 1.20 eq.) in an ice bath. Stir at room temperature for 2 h. Monitor the reaction by TLC. After completion, extract with dichloromethane (30 mL x 3). The combined organic layers are washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a residue. The residue is purified by silica gel column chromatography (PE / EA = 5 / 1 to 2 / 1) to afford compound 1d-8 as a white solid in 86% yield. 1 H NMR(400MHz,cdcl3)δ5.32–5.21(m,1H),4.23(s,1H),4.11(q,J=6.9Hz,1H),1.46(dd,J=12.3,1.4Hz,18H),1.24(t,J=6.4Hz,3H).HRMS(ESI):calcd forC 13 H 25 NO5[M+Na] + 298.1625, found 298.1632.

[1087] (2R)-3-((tert-butoxy(((2R,3S)-4-(tert-butoxy)-3-((tert-butoxycarbonyl)amino)-4-oxobutan-2-yl)oxy)phosphanyl)oxy)propane-1,2-diyldistearate 2d-8

[1088]

[1089] 1d-8 and 4a-8 were treated according to the method described in 2d-1 to obtain the target compound as a colorless transparent oil with a yield of 30%. 1 H NMR(400MHz,Chloroform-d)δ5.40(d,J=9.2Hz,1H),4.98–4.81(m,1H),4.65(dq,J=9.7,5.0Hz,1H),4.26(ddt,J=31.0,11.9,5.1Hz ,4H),2.33(tt,J=8.5,4.1Hz,4H),1.62(t,J=7.3Hz,4H),1.52–1.36(m,27H),1.25(s,59H),0.89(d,J=6.2Hz,6H).HRMS(ESI):calcd for C56 H 108 NO 11 P[M+Mg] + 1025.7505, found 1025.7515.

[1090] (2R)-3-((tert-butoxy(((2R,3S)-4-(tert-butoxy)-3-((tert-butoxycarbonyl)amino)-4-oxobutan-2-yl)oxy)phosphoryl)oxy)propane-1,2-diyldistearate 3d-8

[1091]

[1092] According to the method described in 6a-1, 2d-8 was treated to obtain the target compound, colorless transparent oil, yield 33%. 1 HNMR (400 MHz, Chloroform-d) δ 5.39 (d, J = 9.4 Hz, 1H), 4.98 - 4.82 (m, 1H), 4.65 (dp, J = 9.6, 4.9 Hz, 1H), 4.38 - 3.99 (m, 4H), 2.33 (td, J = 7.5, 3.8 Hz, 4H), 1.61 (t, J = 7.1 Hz, 4H), 1.52 - 1.35 (m, 30H), 1.25 (s, 56H), 0.87 (t, J = 6.7 Hz, 6H). HRMS (ESI): calcd for C 56 H 108 NO 12 P[M+Na] + 1040.7501, found 1040.7503.

[1093] O-(((R)-2,3-bis(stearoyloxy)propoxy)(hydroxy)phosphoryl)-L-threonineC8

[1094]

[1095] According to the method described in A11, 3d-8 was treated to obtain the target compound, light yellow solid, yield 71%. 1H NMR (400 MHz, CDC13 and 20 μL TFA) δ = 7.64 (br s, 3H), 5.07 (br s, 1H), 4.68 (br s, 1H), 4.49-4.34 (m, 2H), 4.33-4.11 (m, 3H), 2.40 (br t, J = 7.6 Hz, 4H), 1.65-1.53 (m, 7H), 1.26 (br s, 56H), 0.89 (br t, J = 6.8 Hz, 6H). 13 C NMR (101 MHz, CDC13 and 20 μL TFA) δ = 176.69, 176.38, 169.18, 73.05, 72.02, 63.04, 62.99, 58.34, 34.11, 29.72, 29.68, 29.39, 29.15, 29.00, 24.69, 22.69, 18.27, 14.08. HRMS (ESI): calcd for C 43 H 84 NO 10 P[M-H] - 804.5760, found 804.5738.

[1096] Biological experimental test example

[1097] Test Example 1 CHO and Beas-2B cell toxicity test

[1098] Principle of experiment: CHO and Beas-2B are normal cells. The toxicity of the compounds prepared in the above examples is detected by detecting the effect on the survival of normal cells after 200 μM administration by CCK-8 method.

[1099] The content of the cell toxicity test is:

[1100] The cells in the logarithmic growth phase were trypsinized, centrifuged and diluted to a certain proportion, and inoculated in 96-well plates (100 μL / well, three replicates) at a concentration of CHO (2 x 10 4 cells / mL), Beas-2B (10 5 cells / mL), and incubated in an incubator at 37 °C, 5% CO2 for 24 h. 200 μM of different types of PS derivatives were added, and the CHO and Beas-2B cells were treated for 24 h, and the number of cell survival was detected by CCK-8 reagent (450 nm). The calculation formula of cell survival rate is as follows:

[1101] Cell survival rate (%) = (ODPS / ODControl) x 100%

[1102] Experimental results: the IC50 As shown in Table 1 below:

[1103] Table 1: Cytotoxicity of PS derivatives to CHO and Beas-2B cells

[1104]

[1105]

[1106] When 200 μM PS was used, the cell survival rate was greater than 50%, which was considered to be the IC 50 >200 μM, otherwise it is considered IC 50 <200

[1107] μM

[1108] Test Example 2 Proliferation Activity of Human Neuroblastoma SH-SY5Y Cells

[1109] The proliferation activity of the compounds prepared in the examples of the present invention on human neuroblastoma SH-SY5Y cells was further investigated. Soybean phosphatidylserine (Soy-PS) was selected as the positive drug, and the experimental conditions for positive and drug determination were consistent.

[1110] The cell proliferation activity test content is:

[1111] The cells in the logarithmic growth phase were digested with trypsin, collected by centrifugation and diluted to a certain ratio, and the concentration was 1.5×10 5 Cells were seeded in 96-well plates (100 μL / well, triplicate wells) at a concentration of 10 cells / mL and cultured in a 37°C, 5% CO2 incubator for 24 hours. Fresh culture medium containing 60 μM PS derivatives was replaced and treated for 48 hours for neuronal cell proliferation assay. Following incubation, CCK-8 (10 μL / well) was added, and after incubation in the dark for 2 hours, the absorbance at 450 nm (OD 450) was measured to calculate cell viability. The cell viability calculation formula is as follows:

[1112] Cell viability (%) = (ODPS / ODControl) × 100%

[1113] Experimental results: The results are shown in Table 2 below:

[1114] Table 2 Proliferative activity of the compounds in the examples on SH-SY5Y cells

[1115] Compound SH-SY5Y proliferationa Compound SH-SY5Y proliferationa A4 12.37±8.48 A23 4.34±3.38 A5 20.25±6.87 A24 29.4±3.28 A6 2.53±7.04 A26 20.01±5.97 A10 6.01±1.84 A27 21.01±3.94 A12 20.16±7.76 A28 5.67±5.05 A13 21.57±3.44 A29 27.97±13.57 A14 7.6±7.58 B3 14.36±5.7 A18 15.44±3.35 B4 13.87±8.39 A19 15.71±4.07 B6 25.9±1.83 A20 10.08±10.4 C6 13.73±3.36 A21 16.55±2.93 Soy-PS 23.64±3.91 A22 23.53±4.49 Blank 0

[1116] a. Increase in proliferation activity (60 μM) relative to that of the blank group.

[1117] Test Example 3 Neuroprotective Effects of Phosphatidylserine Derivatives of the Present Invention

[1118] Further investigation of the neuroprotective effect of phosphatidylserine derivatives, this study through the rotenone (5 μM, 48 h) induced, resulting in about 50% of SH-SY5Y cell death, to establish SH-SY5Y cell Parkinson model, with EPA as a positive control, and the experimental group was given 60 μM different PS derivatives, in this model to evaluate the neuroprotective activity of PS derivatives.

[1119] The steps are as follows:

[1120] The SH-SY5Y cells in the logarithmic growth phase were trypsinized, centrifuged and diluted to 1.5 x 10 5 cells / mL concentration was inoculated in 96-well plates (100 μL / well), and incubated at 37°C, 5% CO incubator in the dark for 24 h. Then, the fresh medium containing different concentrations of PS derivatives was replaced, and incubated in the incubator for 4 h, then Rot solution (final concentration 5 μM, about 50% of the cells were killed) was added, and returned to the incubator for further incubation in the dark for 48 h. After incubation, CCK-8 (10 μL / well) was added in the dark, and incubated in the incubator for 2 h, then the absorbance at 450 nm was measured, and the cell survival rate was calculated, and the calculation method was the same as described before.

[1121] Experimental results: according to the proliferation of SH-SY5Y cells, the compounds were rated, and the rating criteria were:

[1122] D: 0-10;

[1123] C: ≥10 and <20;

[1124] B: ≥20 and <30;

[1125] A: ≥30.

[1126] The results are shown in Table 3 below:

[1127] Table 3 Protection activity detection of example compounds on rotenone-induced SH-SY5Y cell Parkinson model

[1128] Compound SH-SY5Y proliferationa Rating Compound SH-SY5Y proliferationa Rating A4 24.77±3.52 B A26 34.14±7.48 A A5 33.71±3.52 A A27 32.49±5.07 A A6 7.21±2.93 D A28 11.51±6.1 C A7 15.42±4.49 C A29 29.82±4.87 B A8 17.72±2 C A30 18.88±3.85 C A9 15.94±3.39 C B1 15.3±1.82 C A10 21.48±4.24 B B3 18.31±2.93 C A11 19.75±2.26 C B4 17.96±5.8 C A12 14.8±2.92 C B5 38.96±2.56 A A13 16.27±3.55 C B6 38.36±6.95 A A14 14.28±6.52 C B7 26.84±6.4 B A15 13.58±4.32 C C1 16.47±1.57 C A17 17.16±1.62 C C2 17.79±2.51 C A18 10.78±1.9 C C3 17.89±1 C A19 3.94±2.58 D C4 10.59±5.04 C A20 18.93±2.57 C C6 14.81±2.3 C A21 13.24±4.94 C C7 20.35±6.45 B A22 22.34±5.65 B C8 17.76±3.64 C A23 20.7±0.75 B EPA (100 μΜ) 16.66±4.42 C A24 42.48±1.9 A Blank 0

[1129] a. Increase in proliferative activity (60 μM) relative to the blank group.

[1130] In summary, the structure-activity relationship analysis shows that: 1) the unsaturation degree of the fatty acid chain is positively correlated with the activity; 2) the mono-substituted nitro group or fluorobenzoic group at the tail of the fatty acid can significantly enhance the activity, but the double aryl substitution can significantly inhibit the activity; 3) the compounds maintaining the intact glycerol skeleton structure show better biological activity than the amine group modified derivatives; 4) the serine amine group benzoylation modification can lead to reduced activity, while the methylene methylation (threonine structure) has no significant effect on the activity.

[1131] Test Example 4 Effect of Partial Phosphatidylserine Derivatives of the Invention on the Body Weight of Mice

[1132] Experimental materials: SPF level wild type male C57BL / 6J mice (three months old, 28-32 g) were used in the AD model, purchased from Zhejiang Weitong Lihua Experimental Animal Technology Co., Ltd. (License No. SYXK (Shanghai) 2023-0042), and were raised in the SPF level room of the Experimental Animal Center of Shanghai Jiaotong University with a 12 h light / 12 h dark cycle (light source turned on at 7:00 and turned off at 19:00), the temperature was controlled at 22±2℃, and the humidity was controlled at 60±5%. The animal experiments were strictly conducted in accordance with the animal experiment regulations and were approved by the Shanghai Municipal Science and Technology Commission (Ethical Approval No. A2024425).

[1133] Experimental method: The natural soybean-derived PS derivative Soy-PS was used as a positive control, and 42 three-month-old C57BL / 6 male adult mice (28-32 g) were randomly divided into six groups: a blank control group, an Aβ 1–42 injury control group, an Aβ 1–42 +24 compound 25 mg / kg / d group, an Aβ 1–42 +24 compound 50 mg / kg / d group, an Aβ 1–42 +24 compound 100 mg / kg / d group, and an Aβ 1–42 +100 mg / kg / d Soy-PS group (positive control), with 7 mice in each group. The above groups are briefly written as Control group, Aβ group, Aβ+A24 (L) group, Aβ+A24 (M) group, Aβ+A24 (H) group, and Aβ+Soy-PS group. During the drug administration period, the body weight of all experimental mice was monitored weekly, and the body weight data on day 0 was the baseline value of the day before administration. 1–42 The baseline value before injection.

[1134] Experimental results: It can be found from Figure 1 the above that: on the 7th day of administration, the body weights of the mice in each group were significantly different, the Aβ group, the Aβ+Soy-PS group, and the Aβ+A24 (H) group were significantly lower than the Control group, while the Aβ+A24 (L) group had no significant difference from the Control group; on the 21st day of administration, the body weights of the mice in each group had no significant difference. The analysis shows that: ① the Aβ1-42 It can cause short-term weight loss in young mice; ② Low-dose A24 (Aβ+A24(L)) can quickly repair Aβ 1-42 ③ High doses of A24 (Aβ+A24(H)) may increase the initial metabolic burden and fail to quickly repair Aβ 1-42 ④ In long-term observation, young mice showed 1-42 The metabolic effects of sugars can cause weight gain.

[1135] Test Example 5: Some phosphatidylserine derivatives of the present invention improve the cognitive ability of mice

[1136] In this example, the Morris water maze test (MWM) was used to evaluate the effect of the compound of the present invention on improving the spatial learning and memory ability of mice.

[1137] like Figure 2 As shown in the figure, the number of times the mice in the Aβ group crossed the platform was less than that in the Control group ( ## P<0.01). This indicates that the spatial learning and memory abilities of mice in the Aβ group decreased, indicating that the use of Aβ 1–42 The induced AD model was successfully established. Both phosphatidylserine Soy-PS and A24 can improve Aβ 1–42 In the cognitive ability of induced AD mice, on the 6th day of the water maze test, the number of times the Aβ+Soy-PS group crossed the platform was significantly higher than that of the Aβ group (*P<0.05); the number of times the Aβ+A24(L) group crossed the platform was higher than that of the Aβ group (****P<0.0001), and the number of times the A24-administered groups crossed the platform was higher than that of the Aβ group, with significant differences. These results indicate that the phosphatidylserine derivative Soy-PS and A24, especially low-dose A24, can effectively improve the cognitive function of Aβ. 1-42 Induced cognitive impairment.

[1138] Test Example 6 Phosphatidylserine derivatives of the present invention improve neuronal degeneration and synaptic loss in AD mice

[1139] Hematoxylin and eosin staining (HE staining) uses hematoxylin (a basic dye) and eosin (an acidic dye) to stain different components in tissue samples. It can clearly show the differences between the cell nucleus and cytoplasm, so as to distinguish and observe the structure of cells.

[1140] Experimental methods:

[1141] The group with the most pronounced behavioral responses among the three groups treated with Aβ, Aβ+Soy-PS, and Aβ+A24 was selected as the experimental group for HE staining. Three mice were randomly selected from each group. After anesthesia, the mice were rapidly decapitated and brains removed (performed on ice). Fresh tissue from the isolated left hemisphere (three samples per group) was placed in 4% paraformaldehyde and fixed at 4°C for 24 hours. The tissue was then removed from the fixative, and the tissue surrounding the hippocampus was trimmed with a scalpel. The tissue was then dehydrated in a 15% sucrose solution at 4°C until it sank to the bottom, and then transferred to a 30% sucrose solution for dehydration until it sank to the bottom. The dehydrated tissue was then embedded in OCT and, after the OCT bleached and hardened, cut into 8-10 μm sections on a freezing microtome. The tissue was mounted on a slide and stained with hematoxylin for 1-2 minutes and eosin for 2-3 minutes. The sections were then dehydrated with pure alcohol, made transparent with xylene, and finally mounted with neutral gum. The sections were observed under a light microscope, and images of the hippocampus were collected. Five randomly selected fields of view for each sample were quantitatively analyzed using ImageJ software.

[1142] Experimental conclusion:

[1143] The results of HE staining were quantitatively analyzed. Figure 3 As shown in the figure, the cell density of the hippocampus in the A24-treated group was higher than that in the Aβ group, and there was a significant difference (*P<0.05). This indicates that the A24 compound can effectively reverse the Aβ 1-42 Induced hippocampal neuronal degeneration, it can effectively protect the hippocampal synaptic structure and reduce Aβ 1-42 Induced synaptic damage.

[1144] Test Example 7: PS derivatives reduce glial cell activation and inflammatory factor levels

[1145] Studies have shown that excessive activation of microglia can release pro-inflammatory factors such as IL-1β, IL-6, and TNF-α, which can aggravate neurodegeneration by destroying the integrity of the blood-brain barrier, impairing synaptic plasticity, inducing programmed neuronal cell death, and inhibiting neurogenesis in the hippocampus. This example evaluates the neuro-anti-inflammatory effect of the PS derivative A24 and quantitatively analyzes Iba-1 in the hippocampus using immunofluorescence staining. + The density of microglia and the mRNA expression levels of pro-inflammatory factors (IL-1β, IL-6, TNF-α) were detected to systematically elucidate the regulatory effect of A24 on neuroinflammation at the cellular and molecular levels.

[1146] Experimental methods:

[1147] Immunofluorescence:

[1148] Select the Control group, Aβ group, Aβ + Soy-PS group and Aβ + A24 group as the experimental group, and randomly take 3 mice in each group. Immunofluorescence detection was performed on the left hemisphere tissue. Take the left hemisphere of the mouse (3 samples per group), and the steps before freezing sectioning are the same as described in the HE staining. After cutting the tissue into 8-10 μm sections on a freezing microtome, fix the tissue with 4% paraformaldehyde for 20 min. Then, the tissue sections were subjected to antigen retrieval with EDTA antigen retrieval buffer, and BSA blocking solution was added for incubation and blocking for 30 min. Then, the sections were incubated with the primary antibody at 4°C overnight, and then incubated with the corresponding secondary antibody at room temperature for 1 h, DAPI was stained at room temperature for 10 min, and then the sections were mounted with anti-fluorescence quenching mounting medium. Fluorescence microscopy was used to observe and collect images, and 5 fields of each sample were randomly selected for quantitative analysis.

[1149] Fluorescence quantitative RT-PCR:

[1150] First, total RNA was extracted from the mouse hippocampus using Trizol reagent, and then RNA was subjected to reverse transcription reaction using Revert Aid First Strand cDNA Synthesis Kit kit, and then HieffTM qPCR Green Master Mix (No Rox Plus) 11201ES kit was used for cDNA quantification and PCR amplification. Finally, β-actin was used as a reference, and the relative expression of the target gene was calculated by ΔΔCT method. The ΔΔCT method is:

[1151] A = CT (target gene, test) - CT (internal standard gene, test)

[1152] B = CT (target gene, control) - CT (internal standard gene, control)

[1153] K = A - B

[1154] Expression fold = 2 -K

[1155] Experimental results:

[1156] From Figure 4 and Figure 5 it can be seen that immunofluorescence analysis shows that Figure 4 , the number of Iba-1 + microglial cells in the hippocampus of the Aβ model group was significantly increased compared with the blank control group (P < 0.001), showing a typical over-activation state. After the intervention of PS derivative A24, the abnormal activation of microglial cells was significantly inhibited Figure 4 . qPCR detection further confirmed Figure 5), the mRNA expression levels of pro-inflammatory factors TNF-a and IL-6 in the hippocampus of the A24 administration group were significantly lower than those in the Aβ group. These results suggest that the PS derivative A24 may exert a neuroprotective effect by inhibiting Aβ-induced microglial activation and down-regulating the expression of related inflammatory factors. 1-42 induced microglial activation and down-regulate the expression of related inflammatory factors.

[1157] Example 8 Effect of derivative A24 on protein levels in hippocampal tissue

[1158] Experimental methods: Western blot was used to detect the expression changes of key receptors in the cholinergic system CHRM5, PI3K / Akt pathway-related proteins (p-PI3K, p-AKT), and ERK / CREB pathway proteins (p-ERK, ERK, p-CREB), as well as the synaptic plasticity marker synaptophysin (SYN) (with β-actin as the internal reference).

[1159] Experimental results: As shown in Figure 6 compared with the Aβ group, the CHRM5 protein level in the Aβ+A24 group was significantly increased. The WB results verified the reliability of the previous RNA-Seq and qPCR data, confirming that the PS derivative A24 can enhance the function of the cholinergic system by up-regulating the expression of the muscarinic acetylcholine receptor gene Chrm5. At the same time, A24 treatment significantly increased the expression levels of key proteins such as p-PI3K, p-AKT, p-ERK / ERK, p-CREB, and synaptophysin (SYN) in the Aβ+A24 group, indicating that it activated both the ERK / CREB and PI3K / AKT signaling pathways.

[1160] Mechanistically, A24 may, on the one hand, promote CREB phosphorylation by activating the ERK signaling cascade, thereby enhancing the transcription and protein expression of synaptophysin (SYN) gene, improving synaptic plasticity and nerve regeneration in the hippocampus; on the other hand, it may participate in the process of neuronal differentiation and survival by regulating the PI3K / Akt signaling pathway. The synergistic effect of the two pathways improves the cognitive function of AD model mice.

[1161] Example 9 PD test

[1162] Experimental materials: C57BL6 / J male mice (9 weeks old) were selected in this example, and a subacute PD model was established by intraperitoneal injection of 30 mg / kg MPTP for 7 consecutive days. This model can simulate the characteristic damage of dopaminergic neurons in the substantia nigra and motor dysfunction in PD. The mice were randomly divided into 6 groups, 7 in each group: blank control group (normal saline), MPTP modeling group, MPTP+A24 (25 mg / kg) group, MPTP+A24 (50 mg / kg) group, MPTP+A24 (100 mg / kg) group and MPTP+Soy-PS (100 mg / kg) group. The above groups are abbreviated as Control group, MPTP group, MPTP+A24 (L) group, MPTP+A24 (M) group, MPTP+A24 (H) group and MPTP+Soy-PS group. Soy-PS was used as a positive control to systematically evaluate its improvement effect on the motor behavior of PD mice, including motor coordination, gait abnormalities and motor retardation, etc.

[1163] The experimental procedure was divided into two stages: 1) modeling period (days 1-7): except for the blank group, the rest of the groups were intraperitoneally injected with 30 mg / kg MPTP daily; 2) treatment period (days 8-28): after the modeling was successful (i.e., there was a statistically significant difference in behavior between the blank group and the injured group), each treatment group was given the corresponding drug by gavage at 9:00-10:00 daily. The control and model groups were given normal saline. After the behavior test was completed, the mice were sacrificed on day 29, and tissue samples were collected for subsequent analysis.

[1164] Experimental methods:

[1165] Rotarod test: used to systematically evaluate the overall motor coordination and balance of mice. The test time points were set at 0 (verify model success), 7, 14 and 21 days after MPTP modeling success. The test parameters were set as follows: initial rotation speed 10 rpm, linearly accelerated to 40 rpm within 300 s, and the time for the mouse to fall off the rotating rod was recorded (maximum test time 300 s). To ensure data reliability, each mouse was tested 3 times with at least 30 min interval, and the average of the three tests was taken as the motor function index at that time point. Environmental interference factors were strictly controlled during the test to ensure the repeatability of the results.

[1166] Forelimb grip strength test: to evaluate the limb muscle strength of mice. It was performed on day 20 after successful modeling, using a digital grip strength meter: the mouse's both forelimbs were placed on a horizontal pull rod, and the tail was pulled slowly and uniformly (about 1 cm / s) until the mouse's forelimbs were released, and the maximum grip strength value (unit: Newton) displayed by the instrument was recorded. Each mouse was tested for 3 times continuously, with an interval of 5 min to eliminate the influence of fatigue, and the average value of the three tests was taken as the analysis data of the index. The environment was kept quiet during the test, and all tests were completed by the same operator to ensure consistency.

[1167] Open field test: to evaluate the level of neural activity, exploratory behavior and anxiety state of mice, which was performed on day 19 after successful modeling. The experiment used a 500x500x400mm cuboid open field box equipped with a behavior monitoring system. Before the test, the mice were adapted to the experimental environment for 30 min, and then adapted in the open field box for 5 min. In the formal experiment, the mice were placed in the center area of the open field, and their behavior performance in 5 min was recorded, and the motion trajectory was tracked by the video analysis system. Each mouse was tested only once, and the total motion distance was taken as the main observation index for inter-group comparison.

[1168] Test Example 10: Phosphatidylserine derivative of the application improves the motor ability of Parkinson's disease mice

[1169] As shown in Figure 7 , the first rotarod test results showed that the fall latency of mice in the MPTP treatment group was significantly shortened compared with the blank control group, indicating that MPTP successfully induced a model of Parkinson's disease motor dysfunction. In terms of efficacy evaluation: 1) no significant improvement was observed in the MPTP+Soy-PS group after 7 days of drug administration, but the effect was shown after the treatment was prolonged to 21 days; 2) all dose groups of A24 (25, 50, 100 mg / kg) could significantly reverse the motor impairment induced by MPTP, and the last rotarod test residence time was prolonged by 85.5±28.8 seconds (p<0.0001), 70.28±14.7 seconds (p<0.001) and 64.4±30.2 seconds (p<0.001) respectively compared with the model group, among which the 25mg / kg group showed the most obvious improvement trend, but there was no statistical difference among the dose groups.

[1170] In addition, the grip strength test results Figure 8 further confirmed that A24 could effectively improve the decline in motor ability caused by MPTP, and the grip strength of mice in the 100mg / kg group was increased by 32.1±13.6% (p<0.001) compared with the model group, showing the best treatment effect among the PS treatment groups. These results indicate that A24 is significantly superior to Soy-PS in reversing motor dysfunction, and its mechanism of action may involve enhancing muscle strength and improving balance and coordination.

[1171] The results of the open field test shown in Figure 9 The results of the open field test shown in

[1172] In summary, the present application, through systematic optimization and activity evaluation of phosphatidylserine (PS) structure, not only obtains a series of new compounds with significant nerve growth promotion and nerve protection activity, but also further reveals the structure-activity relationship of PS derivatives. These findings provide important theoretical guidance and experimental basis for the development of new drugs for the treatment of neurodegenerative diseases, and have important clinical application value.

[1173] All documents mentioned in this application are incorporated herein by reference as if each individual document were incorporated by reference. In addition, it is to be understood that the application can be carried out by specifically different embodiments and that each disclosed embodiment can be implemented with or without the corresponding benefits disclosed herein.

Claims

1. A compound of formula I, or a tautomer, stereoisomer, or pharmaceutically acceptable salt thereof, in, R1 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 degrees of unsaturation, or a substituent selected from the following group: R2 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 unsaturations or a substituent selected from the following group: R A Selected from the following group: C5-C29 alkyl, C5-C29 alkenyl containing 1-5 degrees of unsaturation, R B is H or 1-2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; X1 or X2 is -O- or -NH-; and X1 and X2 are not both -NH-; X3 is -O- or -NH-; R3 is selected from the group consisting of H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; R4 is H or The additional conditions are: R1 or R2 is not And the compound is not 2. The compound according to claim 1, wherein The compound of formula I is a compound of formula II, in, R1 is a C5-C29 alkenyl group containing 1-5 unsaturations or a substituent selected from the following group: R2 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 unsaturations or a substituent selected from the following group: R A Selected from the following group: C5-C29 alkyl, C5-C29 alkenyl containing 1-5 degrees of unsaturation, R B is H or 1-2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; X1 or X2 is -O- or -NH-; and X1 and X2 are not both -NH-; X3 is -O- or -NH-.

3. The compound according to claim 1, wherein The compound of formula I is a compound of formula II-A, in, R1 is a C5-C29 alkenyl group containing 1-5 unsaturations or a substituent selected from the following group: R2 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 unsaturations or a substituent selected from the following group: R A Selected from the following group: C5-C29 alkyl, C5-C29 alkenyl containing 1-5 degrees of unsaturation, R B is H or 1-2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; X3 is -O- or -NH-; The additional conditions are: When R1 is a C5-C29 alkenyl group containing 1-5 degrees of unsaturation, R2 is not a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 degrees of unsaturation.

4. The compound according to claim 1 or 2, wherein R1 is selected from the group consisting of: and / or R2 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 degrees of unsaturation; Among them, R A 、R B and X3 as defined in claim 1 or 2.

5. The compound according to claim 1 or 2, characterized in that R1 is a C5-C29 alkenyl group containing 1-5 degrees of unsaturation; and / or R2 is selected from the group consisting of: Among them, R A 、R B and X3 as defined in claim 1.

6. The compound according to claim 1, wherein The compound is selected from the group consisting of:

7. The method for preparing the compound according to claim 1, wherein The preparation method of the II-A compound is as follows: (S1) in the presence of a base, the compound of formula 1a is subjected to an esterification reaction with a fatty acid having different or identical R1 and R2 substituents to obtain intermediates 3a-1 to 3a-30; (S2) In an inert solvent, intermediates 3a-1 to 3a-30 are treated with TFA to undergo deprotection reaction to obtain intermediates 4a-1 to 4a-30; (S3) In the presence of an inert solvent and nitrogen, intermediates 4a-1 to 4a-30 are coupled with tert-butyl serine and tert-butyl tetraisopropylphosphorodiamidate to form phosphite triester intermediates 5a-1 to 5a-30; (S4) In an inert solvent, intermediates 5a-1 to 5a-30 are oxidized to obtain intermediates 6a-1 to 6a-30, which are then deprotected to obtain compounds A1 to A30; wherein each group is as described in claim 1.

8. A composition, characterized in that Include: (1) the compound according to claim 1, or its tautomer, stereoisomer, or pharmaceutically acceptable salt; and (2) Pharmaceutically acceptable carrier.

9. A use of a compound of formula I, characterized in that For preparing a drug for preventing and / or treating neurodegenerative diseases; in, R1 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 degrees of unsaturation, or a substituent selected from the following group: R2 is a C5-C29 alkyl group or a C5-C29 alkenyl group containing 1-5 unsaturations or a substituent selected from the following group: R A Selected from the following group: C5-C29 alkyl, C5-C29 alkenyl containing 1-5 degrees of unsaturation, R B is H or 1-2 substituents each independently selected from the group consisting of halogen, -NO2, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; X1 or X2 is -O- or -NH-; and X1 and X2 are not both -NH-; X3 is -O- or -NH-; R3 is selected from the group consisting of H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy; R4 is H or 10. The use according to claim 9, characterized in that The disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, depression, and Huntington's disease.

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