Nitrogen-containing chain compound, preparation method, composition containing nitrogen-containing chain compound and application

Abstract

The invention discloses a nitrogen-containing chain compound, a preparation method, a composition containing the nitrogen-containing chain compound and application of the nitrogen-containing chain compound. The invention specifically discloses a compound I or a pharmaceutically acceptable salt thereof. The LNP preparation prepared from the nitrogen-containing chain compound is relatively uniform in nanoparticle size, relatively high in encapsulation efficiency and relatively high in in-vivo expression activity. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to a nitrogen-containing chain compound, a preparation method, a composition containing the same and applications. Background Art

[0002] Nucleic acid drugs are a key area of ​​current basic and applied research. They can be used to prevent and / or treat viral and bacterial infections, tumors, metabolic diseases, and other diseases. Their lower production costs and shorter production cycles facilitate the rapid development of personalized medicines. However, nucleic acids are negatively charged macromolecules that have difficulty penetrating cell membranes and are also unstable. The development of various nucleic acid packaging and delivery systems can, to some extent, overcome this instability and improve their delivery efficiency.

[0003] Lipid nanoparticles have been demonstrated to be useful as vehicles for delivering bioactive substances (such as small molecule drugs, proteins, and nucleic acids) into cells and / or intracellular compartments. Optimizing nucleic acid drug delivery systems by designing and optimizing the types and dosages of the various components within lipid nanoparticles is crucial for enhancing the efficacy of nucleic acid drugs for prevention and treatment. This is particularly true for lipid compounds that can be used to deliver RNA prophylactic and / or therapeutic agents, as well as related methods and compositions.

[0004] Given the importance of ionizable lipid compounds that can be used to deliver nucleic acid drugs, the development of novel structural ionizable lipid compounds is urgently needed. Summary of the Invention

[0005] The present invention aims to provide a novel ionizable lipid compound that can be used to deliver nucleic acid drugs, thereby expanding the variety of ionizable lipid compounds and the selection of delivery vehicles for nucleic acid prophylactic and / or therapeutic agents. To address the above technical problems, the present invention provides a nitrogen-containing chain compound, a preparation method, a composition containing the same, and applications thereof. The LNP preparations prepared using the nitrogen-containing chain compound of the present invention exhibit relatively uniform nanoparticle size, high encapsulation efficiency, and high in vivo expression activity.

[0006] The present invention provides a compound I or a pharmaceutically acceptable salt thereof,

[0007]

[0008] Among them, R 1 is a C substituted by one or more hydroxyl groups 1-6 alkyl;

[0009] X and Y are independently C 1-12 alkylene;

[0010] Z 1 and Z 2 Independently

[0011] W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0012] R 2 for R 2a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-6 Alkyl; R 2b C 1-6 Alkylene, R 2c C 1-15 alkyl;

[0013] R 3 C 1-15 Alkyl or R 3a C 1-6 Alkylene, R 3b C 1-15 alkyl;

[0014] R 4 H, C 1-15 alkyl, or -C 3-10 Cycloalkyl-C 1-15 Alkyl; R 4a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa ; R 4aa H or C 1-6 alkyl;

[0015] R 5 C 1-15 Alkyl or R 5a C 1-6 Alkylene, R 5b C 1-15 alkyl.

[0016] In certain preferred embodiments of the present invention, certain groups of the compound I or a pharmaceutically acceptable salt thereof are defined as follows, and the unmentioned groups are the same as those described in any embodiment of the present invention (referred to as "in a certain embodiment of the present invention").

[0017] In one embodiment of the present invention, R 1is a C substituted by one or more hydroxyl groups 1-6 alkyl;

[0018] X and Y are independently C 1-12 alkylene;

[0019] Z 1 and Z 2 Independently

[0020] W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0021] R 2 for R 2a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-6 alkyl;

[0022] R 3 C 1-15 Alkyl or R 3a C 1-6 Alkylene, R 3b C 1-15 alkyl;

[0023] R 4 H, C 1-15 alkyl, or -C 3-10 Cycloalkyl-C 1-15 Alkyl; R 4a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa ; R 4aa H or C 1-6 alkyl;

[0024] R 5 C 1-15 Alkyl or R 5a C 1-6 Alkylene, R 5b C 1-15 alkyl.

[0025] In one embodiment of the present invention, R 1 、R2aa and R 4aa wherein the C 1-6 Alkyl and C 1-6 The "C" in the alkyl group 1-6 "alkyl" are each independently C 1-4 Alkyl, for example methyl, ethyl, n-propyl or n-butyl.

[0026] In one embodiment of the present invention, R 1 、R 2aa and R 4aa wherein the C 1-6 Alkyl and C 1-6 The "C" in the alkyl group 1-6 "Alkyl" is each independently a straight chain or branched C 1-6 alkyl.

[0027] In one embodiment of the present invention, X, Y, R 2a and R 4a In the C 1-12 Alkylene, -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa 、-C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa "C" in 1-12 "Alkylene" are each independently C 3-7 Alkylene, e.g.

[0028] In one embodiment of the present invention, X, Y, R 2a and R 4a In the C 1-12 Alkylene, -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa 、-C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa "C" in 1-12 "Alkylene" is each independently a straight chain or branched C 1-12 Alkylene.

[0029] In one embodiment of the present invention, W 1 、W 2 、R 3a and R 5a In the C 1-6 Each alkylene group is independently a methylene group, -CH2CH2-,

[0030]

[0031] In one embodiment of the present invention, R 2b In the C 1-6 Alkylene is C 1-3 Alkylene groups, such as methylene.

[0032] In one embodiment of the present invention, W 1 、W 2 、R 3a 、R 5a and R 2b In the C 1-6 Each alkylene group is independently a methylene group, -CH2CH2-,

[0033]

[0034] In one embodiment of the present invention, W 1 、W 2 、R 3a and R 5a In the C 1-6 The alkylene groups are each independently linear or branched C 1-6 Alkylene.

[0035] In one embodiment of the present invention, R 2b In the C 1-6 Alkylene is a straight chain or branched C 1-6 Alkylene.

[0036] In one embodiment of the present invention, W 1 、W 2 、R 3a 、R 5a and R 2b In the C 1-6 The alkylene groups are each independently linear or branched C 1-6 Alkylene.

[0037] In one embodiment of the present invention, R 2a 、R 3 、R 3b 、R 4 、R 4a 、R 5 and R 5b In the C 1-15 Alkyl, -C 3-10 Cycloalkyl-C 1-15 The "C" in the alkyl group 1-15 Alkyl" is C 3-12 Alkyl, such as n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0038]

[0039] In one embodiment of the present invention, R 2c In the C 1-15 Alkyl is C 3-12 Alkyl, for example n-pentyl or n-heptyl.

[0040] In one embodiment of the present invention, R 2a 、R 3 、R 3b 、R 4 、R 4a 、R 5 、R 5b and R 2c In the C 1-15 Alkyl, -C 3-10 Cycloalkyl-C 1-15 The "C" in the alkyl group 1-15 Alkyl" is C 3-12 Alkyl, such as n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0041]

[0042] In one embodiment of the present invention, R 2a 、R 3 、R 3b 、R 4 、R 4a 、R 5 and R 5b In the C 1-15 Alkyl, -C 3-10 Cycloalkyl-C 1-15 The "C" in the alkyl group 1-15 "Alkyl" is a straight or branched chain C 1-15 alkyl.

[0043] In one embodiment of the present invention, R 2c In the C 1-15 Alkyl is a straight chain or branched C 1-15 alkyl.

[0044] In one embodiment of the present invention, R 2a 、R 3 、R 3b 、R 4 、R 4a 、R 5 、R 5b and R 2c In the C 1-15 Alkyl, -C 3-10 Cycloalkyl-C 1-15 The "C" in the alkyl group 1-15 "Alkyl" is a straight or branched chain C 1-15 alkyl.

[0045] In one embodiment of the present invention, R 2a In the C 1-15 Alkyl is C 3-10 Alkyl, such as n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0046] In one embodiment of the present invention, R 2a In the C 1-15 Alkyl is C 3-10 Alkyl, such as n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0047] In one embodiment of the present invention, R 2a In the C 1-15 Alkyl is a straight chain or branched C 1-15 alkyl.

[0048] In one embodiment of the present invention, R 2a and R 4a In the C 1-15 Alkenyl is C 3-10 Alkenyl, e.g.

[0049] In one embodiment of the present invention, R 2a and R 4a In the C 1-15 Alkenyl is a straight chain or branched C 1-15 Alkenyl.

[0050] In one embodiment of the present invention, R 2a and R 4a In the -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa and -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa "C" in 3-6 "Cycloalkyl" is cyclopropyl or cyclobutyl.

[0051] In one embodiment of the present invention, R 4 In the -C 3-10 Cycloalkyl-C 1-15 The "C" in the alkyl group 3-10 "Cycloalkyl" is C 3-6 Cycloalkyl, for example cyclohexyl.

[0052] In one embodiment of the present invention, R 1 C substituted by a hydroxyl group 1-6 alkyl.

[0053] In one embodiment of the present invention, R 1 for

[0054] In one embodiment of the present invention, R 1 In the case of a hydroxyl group, only the terminal carbon not connected to the nitrogen atom is replaced by a hydroxyl group.

[0055] In one embodiment of the present invention, X and Y are independently C 3-7 Alkylene.

[0056] In one embodiment of the present invention, X and Y are independently

[0057] In one embodiment of the present invention, Z 1 for *Indicates the same as W 1 connect.

[0058] In one embodiment of the present invention, Z 2 for *Indicates the same as W 2 connect.

[0059] In one embodiment of the present invention, Z 2 for

[0060] In one embodiment of the present invention, Z 2 for Preferably, Z 2 for *Indicates the same as W 1 connect.

[0061] In one embodiment of the present invention, W1 and W2 are independently a chemical bond, a methylene group, -CH2CH2-,

[0062] In one embodiment of the present invention, R 2 for

[0063] In one embodiment of the present invention, R 2 for R 2a C 3-10 Alkyl, C 3-10 Alkenyl or -C 3-7 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-3 Alkyl; R 2b C 1-3 Alkylene, R 2c C3-10 alkyl.

[0064] In one embodiment of the present invention, R 2b C 1-3 Alkylene, R 2c C 3-10 alkyl.

[0065] In one embodiment of the present invention, R 2b For methylene.

[0066] In one embodiment of the present invention, R 2c for

[0067] In one embodiment of the present invention, R 2a C 3-10 Alkyl, C 3-10 Alkenyl or -C 3-7 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-3 alkyl.

[0068] In one embodiment of the present invention, R 2a is n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0069]

[0070] In one embodiment of the present invention, R 2a for In one embodiment of the present invention, R 2a is n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0071]

[0072] In one embodiment of the present invention, R 2 for

[0073] In one embodiment of the present invention, R 2 for

[0074] In one embodiment of the present invention, R 2 for

[0075] In one embodiment of the present invention, R 3 C 4-12 Alkyl or R 3a C1-3 Alkylene, R 3b C 6-12 alkyl.

[0076] In one embodiment of the present invention, R 3 for

[0077] In one embodiment of the present invention, R 3 for In one embodiment of the present invention, R 3 for

[0078]

[0079] In one embodiment of the present invention, R 3a For methylene.

[0080] In one embodiment of the present invention, R 3b It is n-heptyl.

[0081] In one embodiment of the present invention, R 3b for In one embodiment of the present invention, R 3b For n-heptyl, In one embodiment of the present invention, for

[0082]

[0083] In one embodiment of the present invention, for

[0084]

[0085] In one embodiment of the present invention, for

[0086]

[0087] In one embodiment of the present invention, R 4 H, C 3-10 alkyl, or -C 3-6 Cycloalkyl-C 3-10 Alkyl; R 4a C 3-10 Alkyl, C 3-10 Alkenyl or -C 3-7Alkylene-C 3-6 Cycloalkyl-R 4aa ; R 4aa H or C 1-3 alkyl.

[0088] In one embodiment of the present invention, R 4a For n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0089] In one embodiment of the present invention, R 4a for

[0090] In one embodiment of the present invention, R 4a For n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,

[0091] In one embodiment of the present invention, R 4 For H,

[0092]

[0093] In one embodiment of the present invention, R 4 for

[0094] In one embodiment of the present invention, R 4 For H,

[0095] In one embodiment of the present invention, R 5 C 6-10 Alkyl or R 5a C 1-3 Alkylene, R 5b C 6-10 In one embodiment of the present invention, R 5a For methylene.

[0096] In one embodiment of the present invention, R 5b It is n-heptyl.

[0097] In one embodiment of the present invention, R 5b It is n-octyl.

[0098] In one embodiment of the present invention, R 5b It is n-heptyl or n-octyl.

[0099] In one embodiment of the present invention, R 5 for

[0100] In one embodiment of the present invention, R 5 for In one embodiment of the present invention, R 5 for

[0101] In one embodiment of the present invention, for

[0102]

[0103] In one embodiment of the present invention, for

[0104] In one embodiment of the present invention, for

[0105]

[0106]

[0107] In one embodiment of the present invention, R 1 is a C substituted by one or more hydroxyl groups 1-3 Alkyl; X and Y are independently C 3-8 Alkylene, preferably C 4-6 Alkylene, such as C5 alkylene;

[0108] Z 1 for *Indicates the same as W 1 connect;

[0109] Z 2 for *Indicates the same as W 2 connect;

[0110] W 1 and W 2 Independently C 1-3 alkylene groups, such as methylene;

[0111] R 2 for R 2a C 3-12 Alkyl; R 2a Preferably C 5-9 Alkyl, such as C7 alkyl; R 3 C 5-15 Alkyl; R3 Preferably C 8-12 Alkyl groups, such as C 10 alkyl;

[0112] R 4 for R 4a C 3-12 Alkyl; R 4a Preferably C 5-9 Alkyl, such as C7 alkyl; R 5 C 5-15 Alkyl; R 5 Preferably C 8-12 Alkyl groups, such as C 10 alkyl.

[0113] In one embodiment of the present invention, R 1 is a C substituted by one or more hydroxyl groups 1-6 alkyl;

[0114] X and Y are independently C 1-12 alkylene;

[0115] Z 1 for *Indicates the same as W 1 connect;

[0116] Z 2 for *Indicates the same as W 2 connect;

[0117] W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0118] R 2 for R 2a C 1-15 Alkyl or C 1-15 alkenyl;

[0119] R 3 C 1-15 alkyl;

[0120] R 4 C 1-15 Alkyl or R 4a C 1-15 Alkyl or C 1-15 alkenyl;

[0121] R 5 C 1-15 alkyl.

[0122] In one embodiment of the present invention, R1 is a C substituted by one or more hydroxyl groups 1-6 alkyl;

[0123] X and Y are independently C 1-12 alkylene;

[0124] Z 1 for *Indicates the same as W 1 connect;

[0125] Z 2 for *Indicates the same as W 2 connect;

[0126] W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0127] R 2 for R 2a C 1-15 Alkyl or C 1-15 alkenyl;

[0128] R 3 C 1-15 Alkyl or R 3a C 1-6 Alkylene, R 3b C 1-15 Alkyl; R 4 C 1-15 Alkyl or R 4a C 1-15 Alkyl or C 1-15 alkenyl;

[0129] R 5 C 1-15 Alkyl or R 5a C 1-6 Alkylene, R 5b C 1-15 alkyl.

[0130] In one embodiment of the present invention, the compound I has the following structure:

[0131]

[0132] Among them, R 1 、R 2 、R 3 、R 4 、R 5 、W 1 、W2 , X and Y are as defined in any of the previous schemes; preferably, the compound I has the following structure:

[0133]

[0134] Among them, R 1 、R 2a 、R 3 、R 4a 、R 5 、W 1 、W 2 , X and Y are as defined in the previous embodiment. In one embodiment of the present invention, the compound I has the following structure:

[0135]

[0136] Among them, R 1 、R 2 、R 3 、R 4 、R 5 、W 1 、W 2 , X and Y are as defined in any of the previous embodiments. In one embodiment of the present invention, the compound I is any of the following structures:

[0137]

[0138] Among them, R 1 、R 2 、R 3 、R 4 、R 5 、W 1 、W 2 、Z 1 、Z 2 , X and Y are as defined in any of the previous embodiments. In one embodiment of the present invention, the compound I is any of the following structures:

[0139]

[0140]

[0141]

[0142]

[0143]

[0144]

[0145] In one embodiment of the present invention, the compound I is any of the following structures:

[0146]

[0147]

[0148] The present invention also provides a use of the compound I or a pharmaceutically acceptable salt thereof in the preparation of a delivery vector for a nucleic acid preventive agent and / or therapeutic agent;

[0149] The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small hairpin RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA) or messenger RNA (mRNA), preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 spike protein (Spike) mRNA, herpes zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0150] The present invention also provides a composition comprising a substance Z, wherein the substance Z is the compound I as described above or a pharmaceutically acceptable salt thereof.

[0151] The present invention also provides a lipid carrier, which includes a substance Z, wherein the substance Z is the aforementioned compound I or a pharmaceutically acceptable salt thereof.

[0152] In one embodiment of the present invention, the lipid carrier further comprises a diluent, which can be a phosphate buffer or a Tris buffer.

[0153] In one embodiment of the present invention, the lipid carrier further comprises phospholipids.

[0154] In one embodiment of the present invention, the phospholipid may be a conventional phospholipid in the art, which is an amphiphilic auxiliary molecule that helps the fusion of lipid particles and cell membranes. The phospholipid may be a phospholipid molecule having an electrically charged polar end and a non-polar end of a fatty chain, such as distearoylphosphatidylcholine (DSPC), dimyristoylphosphocholine (DMPC), dioleoylphosphocholine (DOPC), palmitoylphosphocholine (DPPC), 1,2-distearoylphosphocholine (DSPC), heneicosanoylphosphocholine (DUPC) or palmitoylphosphocholine (POPC).

[0155] In one embodiment of the present invention, the lipid carrier further comprises PEG lipid (polyethylene glycol-modified lipid).

[0156] In one embodiment of the present invention, the PEG lipid may be a lipid molecule having a polyethylene glycol hydrophilic end modified. The PEG lipid is preferably selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol, such as PEG-modified dimyristoylglycerol (DMG-PEG2000).

[0157] In one embodiment of the present invention, the lipid carrier further comprises sterol.

[0158] In one embodiment of the present invention, the sterol may be a conventional sterol in the art, including animal, plant or fungal sterols. The sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid and α-tocopherol, such as cholesterol.

[0159] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the sterol is 0.5-5:1, preferably 0.5-3:1, for example 1.3:1.

[0160] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 2-8:1, such as 3-6:1.

[0161] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 1-10:1, for example 5:1.

[0162] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the PEG lipid is 10-100:1, preferably 10-50:1, for example 33.3:1.

[0163] In the present invention, molar content means the percentage of a substance in the total mass of the lipid carrier, and the sum of the molar contents of the components in the lipid carrier does not exceed 100 mol%.

[0164] In one embodiment of the present invention, the molar content of the substance Z in the lipid carrier is 30 mol% to 70 mol%, for example, 50 mol%.

[0165] In one embodiment of the present invention, the molar content of the substance Z in the lipid carrier is 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0166] In one embodiment of the present invention, the molar content of the phospholipid in the lipid carrier is 5 mol% to 20 mol%, for example, 10 mol%.

[0167] In one embodiment of the present invention, the molar content of the phospholipid in the lipid carrier is 0 mol% to 30 mol%, for example, 0 mol%, 5 mol%, 10 mol%, 15 mol% or 20 mol%.

[0168] In one embodiment of the present invention, the molar content of the sterol in the lipid carrier is 20 mol% to 60 mol%, for example, 38.5 mol%.

[0169] In one embodiment of the present invention, the molar content of the sterol in the lipid carrier is 28.5 mol%, 33.5 mol%, 37.0 mol%, 38.0 mol%, 38.5 mol%, 39.0 mol%, 39.5 mol%, 43.5 mol%, 48.5 mol% or 53.5 mol%.

[0170] In one embodiment of the present invention, the molar content of the PEG lipid in the lipid carrier is about 0.2 mol% to 5 mol%, for example, 1.5% mol.

[0171] In one embodiment of the present invention, the molar content of the PEG lipid in the lipid carrier is 0.5% mol, 1.0% mol, 1.5% mol, 2.0% mol or 3.0% mol.

[0172] In a certain embodiment of the present invention, in the lipid carrier, the lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.

[0173] In one embodiment of the present invention, the lipid carrier is any one of the following formulations:

[0174]

[0175]

[0176] ; Wherein, the substance Z is preferably compound 6 or a pharmaceutically acceptable salt thereof (the structure of compound 6 is as follows);

[0177]

[0178] The phospholipid is preferably DSPC; the sterol is preferably cholesterol; the PEG lipid is preferably DMG-PEG2000; preferably, the lipid carrier is any one of the following formulas:

[0179] prescription Compound 6 or a pharmaceutically acceptable salt thereof: DSPC: cholesterol: DMG-PEG2000 1 35：10：53.5：1.5 2 40：10：48.5：1.5 3 45：10：43.5：1.5 4 50：10：38.5：1.5 5 55：10：33.5：1.5 6 60：10：28.5：1.5 7 50：0：48.5：1.5 8 50：5：43.5：1.5 9 50：15：33.5：1.5 10 50：20：28.5：1.5 11 50：10：39.5：0.5 12 50：10：39.0：1.0 13 50：10：38.0：2.0 14 50：10：37.0：3.0 .

[0180] The present invention also provides a use of the lipid carrier in preparing a delivery vector for a nucleic acid preventive agent and / or therapeutic agent;

[0181] The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small hairpin RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA) or messenger RNA (mRNA), preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 spike protein (Spike) mRNA, herpes zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0182] The present invention also provides a lipid nanoparticle comprising a nucleic acid preventive agent and / or therapeutic agent and the aforementioned lipid carrier;

[0183] The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small hairpin RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA) or messenger RNA (mRNA), preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 spike protein (Spike) mRNA, herpes zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0184] In one embodiment of the present invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles may be (2-30):1. The nitrogen-to-phosphorus ratio refers to the ratio of the moles of ionizable nitrogen atoms in one or more ionizable lipid compounds to the moles of phosphate groups in the RNA. In this application, it refers to the ratio of the moles of ionizable nitrogen atoms in the ionizable lipid nanoparticles to the moles of phosphate groups in the mRNA in the pharmaceutical composition. Preferably, the nitrogen-to-phosphorus ratio is (2-20):1, preferably (3-20):1, for example (3-16):1.

[0185] In one embodiment of the present invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles is 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or 9:1.

[0186] In one embodiment of the present invention, the particle size (average particle size) of the lipid nanoparticles is 10-200 nm, preferably 40-150 nm, and more preferably 50-130 nm, for example, 57.29 nm, 61.55 nm, 69.72 nm, 70.08 nm, 76.03 nm, 89.89 nm, 97.55 nm, 107.37 nm or 119.10 nm.

[0187] In one embodiment of the present invention, the particle size of the lipid nanoparticles is 63.36 mm, 56.53 mm, 71.25 mm, 57.66 mm, 60.92 mm, 55.68 mm, 56.19 mm, 55.52 mm, 67.96 mm, 69.51 mm or 79.13 mm.

[0188] In one embodiment of the present invention, the particle size of the lipid nanoparticles is 57.29 nm, 61.55 nm, 69.72 nm, 70.08 nm, 76.03 nm, 89.89 nm, 97.55 nm, 107.37 nm, 119.10 nm, 63.36 mm, 56.53 mm, 71.25 mm, 57.66 mm, 60.92 mm, 55.68 mm, 56.19 mm, 55.52 mm, 67.96 mm, 69.51 mm or 79.13 mm.

[0189] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.001-0.3, for example, 0.035, 0.074, 0.090, 0.097, 0.118, 0.166 or 0.269.

[0190] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.065, 0.074, 0.062, 0.106, 0.089, 0.108, 0.090, 0.078, 0.085, 0.063 or 0.084.

[0191] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.035, 0.074, 0.090, 0.097, 0.118, 0.166, 0.269, 0.065, 0.074, 0.062, 0.106, 0.089, 0.108, 0.090, 0.078, 0.085, 0.063 or 0.084.

[0192] In one embodiment of the present invention, the encapsulation efficiency of the lipid nanoparticles is 90%-100%, for example, 96.73%, 98.07%, 96.90%, 97.71%, 96.36%, 95.02%, 92.94%, 94.44%, 95.23% or 94.63%.

[0193] In one embodiment of the present invention, the encapsulation efficiency of the lipid nanoparticles is 95.83%, 94.95%, 96.00%, 96.62%, 94.28%, 96.56%, 96.04%, 95.94%, 98.33%, 95.38% or 96.78%.

[0194] In one embodiment of the present invention, the encapsulation efficiency of the lipid nanoparticles is 96.73%, 98.07%, 96.90%, 97.71%, 96.36%, 95.02%, 92.94%, 94.44%, 95.23%, 94.63%, 95.83%, 94.95%, 96.00%, 96.62%, 94.28%, 96.56%, 96.04%, 95.94%, 98.33%, 95.38% or 96.78%.

[0195] In a certain embodiment of the present invention, in the lipid nanoparticles, the lipid carrier encapsulates the nucleic acid preventive agent and / or therapeutic agent.

[0196] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0197] The reagents and raw materials used in the present invention are commercially available.

[0198] The positive progress of the present invention is that the LNP preparation prepared by using the nitrogen-containing chain compound of the present invention has relatively uniform nanoparticle size, high encapsulation efficiency and high in vivo expression activity.

[0199] Unless otherwise specified, the terms used in this invention may be defined as follows:

[0200] It will be understood by those skilled in the art that the structural formulas used in the present invention to describe groups are based on the conventions used in the art. It means that the corresponding group is connected to other fragments and groups in the compound through this site.

[0201] As used herein, a substituent may be preceded by a single dash "-" to indicate that the named substituent is bonded to the parent moiety through a single bond.

[0202] When one of the variables is selected from a chemical bond, it means that the two groups it connects are directly connected.

[0203] When any variable appears multiple times in the definition of a compound, the definition of the variable at each position is independent of the definition at the remaining positions, and their meanings are independent of each other and do not affect each other.

[0204] The term "plurality" refers to 2, 3 or 4.

[0205] The term "alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, and the like.

[0206] The term "alkylene" refers to a straight or branched divalent hydrocarbon group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkylene include, but are not limited to, methylene, ethylene, propylene, 1-methylpropylene, butylene, and the like.

[0207] The term "alkenyl" refers to a straight or branched hydrocarbon group having a specified number of carbon atoms (e.g., C2-C4) and one or more double bonds. The one or more carbon-carbon double bonds may be internal or terminal.

[0208] The term "cycloalkyl" refers to a saturated cyclic group having a specified number of ring carbon atoms (e.g., C3-C8) and the ring atoms consisting only of carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0209] The term "pharmaceutically acceptable salt" refers to salts prepared from compounds of the present invention with relatively nontoxic, pharmaceutically acceptable acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent.

[0210] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0211] The term "prevention" refers to a period of time in which a subject remains healthy relative to a disease or condition mentioned herein. It should be understood that the period depends on the amount of the drug compound that has been administered and the individual factors of the subject discussed elsewhere in this specification. It should be understood that prevention may not be effective in all subjects treated with the compounds according to the present invention. However, the term requires that a statistically significant portion of a cohort or group of subjects is preferably effectively prevented from suffering from a disease or condition or its associated symptoms as referred to herein. Preferably, in this case, it is envisioned that a group or group of subjects would typically develop a disease or condition as referred to herein without taking preventive measures according to the present invention. Those skilled in the art can use various well-known statistical evaluation tools discussed elsewhere in this specification to immediately determine whether a portion is statistically significant.

[0212] When any variable is used to connect two or more groups, those skilled in the art will understand that the actual meaning of the variable is the embodiment obtained without changing or losing one or more hydrogen atoms based on the definition of the variable in the claims and / or the specification; for example, in this application, R 2a A definition of "-C 1-12 Alkylene-C 3-10 Cycloalkyl-R 2aa ”, where “C 1-12 "Alkylene" is used to connect two groups, then its definition is "C 1-12 "Alkylene" itself is an embodiment, and its examples include but are not limited to methylene, ethylene, etc.; and "C 3-10 Cycloalkyl" is used to connect "C 1-12 Alkylene" and R 2aa , when R 2aa When it is not hydrogen, the "C3-C 10 The actual meaning of "cycloalkyl" is "C3-C 10 The structure obtained by "cycloalkyl" losing a hydrogen, that is, "C3-C 10 Cycloalkylene", e.g. When R 2aa When it is hydrogen, the "C3-C 10 The definition of "cycloalkyl" itself is an embodiment, and its examples include but are not limited to wait. BRIEF DESCRIPTION OF THE DRAWINGS

[0213] Figure 1 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 2, compound 3 and compound 6;

[0214] Figure 2is the chemiluminescence intensity at the administration site of mice after intramuscular administration of compound 2, compound 3 and compound 6;

[0215] Figure 3 is the chemiluminescence intensity of the mouse liver area after intramuscular administration of compound 2, compound 3 and compound 6;

[0216] Figure 4 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 8 and compound 9;

[0217] Figure 5 is the chemiluminescence intensity at the administration site of mice after intramuscular administration of compound 8 and compound 9;

[0218] Figure 6 is the chemiluminescence intensity of the mouse liver area after intramuscular administration of compound 8 and compound 9;

[0219] Figure 7 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of SM102, compound 5 and compound 28;

[0220] Figure 8 is the chemiluminescence intensity at the administration site of mice after intramuscular administration of compound 5 and compound 28;

[0221] Figure 9 is the chemiluminescence intensity of the mouse liver area after intramuscular administration of compound 5 and compound 28;

[0222] Figure 10 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 23, compound 24, compound 25, compound 26, compound 27, compound 29 and compound 34;

[0223] Figure 11 is the chemiluminescence intensity at the administration site of mice after intramuscular administration of SM102, compound 23, compound 24, compound 25, compound 26, compound 27, compound 29 and compound 34;

[0224] Figure 12 is the chemiluminescence intensity of the mouse liver area after intramuscular administration of SM102, compound 23, compound 24, compound 25, compound 26, compound 27, compound 29 and compound 34;

[0225] Figure 13 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 1, compound 7, compound 21, compound 32, compound 48, compound 55 and compound 57;

[0226] Figure 14The expression of VZV antigen protein in vivo after the mice were injected with each VZV mRNA LNP via tail vein in Example 16 (3 hours after administration, n=5);

[0227] Figure 15 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 11, compound 19, compound 20, compound 22, compound 30, compound 31, compound 52, compound 58 and compound 60;

[0228] Figure 16 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 33, compound 35, compound 36, compound 37, compound 47, compound 49, compound 50, compound 62, compound 63, compound 64, compound 65, compound 66, compound 67 and compound 68;

[0229] Figure 17 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of LNP1, LNP2, LNP3, LNP4, LNP5, LNP6, LNP7, LNP8, LNP9, LNP10, LNP11, LNP12, LNP13 and LNP14 under different lipid formulations;

[0230] Figure 18 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of NP3, NP4, NP5, NP6, NP7, NP8 and NP9 at different nitrogen and phosphorus ratios;

[0231] Figure 19 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of Compound 10, Compound 12, Compound 13, Compound 14, Compound 15, Compound 16, Compound 17 and Compound 61. DETAILED DESCRIPTION

[0232] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0233] In the examples of the present application, RT in LCMS is the abbreviation of "retention time", and its unit is "minute".

[0234] Preparation Example 1 Preparation of Compound 2

[0235] Step 1: Preparation of 2-1

[0236] Reaction formula:

[0237]

[0238] Material ratio:

[0239] Material Name Molecular weight Feed ratio Feeding amount mmol Decyl epoxide 156 1eq 4.2g 27 7-Bromoheptanoic acid 209 1.1 eq 6.2g 30 Ferric chloride 162 0.025 eq 220mg 1.35 Pyridine 79 0.0125eq 53mg 0.68

[0240] Operation process:

[0241] 7-Bromoheptanoic acid, decyl oxide, ferric chloride, and pyridine were added to a 250 mL reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 4:1) indicated completion of the reaction (product rf value 0.5). Purification by column chromatography yielded 7.4 g of a colorless oil.

[0242] Step 2: Preparation of 2-2

[0243] Reaction formula:

[0244]

[0245] Material ratio:

[0246] Material Name Molecular weight Feed ratio Feeding amount mmol 2-1 365.35 1eq 6.4g 17.5 bitter 144 1.1 eq 2.8g 19.2 EDCI 192 2 eq 6.7g 35 DMAP 122 0.2 eq 420mg 3.5 dichloromethane - - 80mL -

[0247] Operation process:

[0248] To the reaction flask, 2-1, octanoic acid, EDCI, DMAP, and dichloromethane were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 6 g of a colorless oil.

[0249] Step 3: Preparation of compound 2

[0250] Reaction formula:

[0251]

[0252] Material ratio:

[0253] Material Name Molecular weight Feed ratio Feeding amount mmol 2-2 491.55 1.1eq 4.2g 8.55 LQ001-2 441.74 1 eq 3.4g 7.77 K2CO3 138 1.5eq 1.6g 11.6 KI 166 1.5eq 1.9g 11.6 Acetonitrile - - 40mL -

[0254] Operation process:

[0255] 2-2, LQ001-2, K2CO3, KI, and acetonitrile were added to the reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was filtered, concentrated, and purified by column chromatography to yield 3 g of a colorless oil.

[0256] 1H NMR (600MHz, Chloroform-d) δ5.08 (qd, J=6.9, 3.3Hz, 1H), 4.85 (p, J=6.2Hz, 1H), 4.21 (dd, J=11.8, 3.3Hz, 1H), 4.01 (dd, J=11.8, 6.8Hz, 1H),3.69–3.59(m,2H),2.66(d,J=75.2Hz,4H),2.28(dt,J=14.1,7.5Hz,6H),1.65–1.45(m,16H),1.36–1.20(m,56H),0.91–0.84(m,12H).

[0257] MS (ES+) m / z): 852.7 (M+H) + .

[0258] Compound 2 can also be prepared by the following method:

[0259] Step 1: Preparation of 2-1

[0260] Reaction formula:

[0261]

[0262] Material ratio:

[0263] Material Name Molecular weight Feed ratio Feeding amount mmol 7-Bromoheptanoic acid 209 1eq 21g 100 1,2-Epoxydecane 156 1eq 15.6g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mL 2.5

[0264] Operation process:

[0265] 7-Bromoheptanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 17 g of an oil.

[0266] Step 2: Preparation of 2-2

[0267] Reaction formula:

[0268]

[0269] Material ratio:

[0270] Material Name Molecular weight Feed ratio Feeding amount mmol 2-1 365.35 1eq 7.3g 20 Octanoic acid 144.21 1 eq 2.9g 20 EDCI 192 1.5eq 5.7g 30 DMAP 122 0.15 eq 370mg 3 DCM - - 100mL -

[0271] Operation process:

[0272] 2-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 100 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 6.5 g of an oil.

[0273] Step 3: Preparation of 2-3

[0274] Reaction formula:

[0275]

[0276] Material ratio:

[0277] Material Name Molecular weight Feed ratio Feeding amount mmol 8-Bromooctanoic acid 223 1eq 7g 30 9-Heptadecanol 256 1 eq 7.7g 30 EDCI 192 1.5eq 5.8g 45 DMAP 122 0.15 eq 400mg 4.5 DCM - - 100mL -

[0278] Operation process:

[0279] 8-Bromooctanoic acid, 9-heptadecanol, EDCI, DMAP, and DCM were added to a reaction flask and allowed to react at room temperature for 2 h. The product exhibited an Rf value of 0.5 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 10.8 g of an oil.

[0280] Step 4: Preparation of 2-4

[0281] Reaction formula:

[0282]

[0283] Material ratio:

[0284] Material Name Molecular weight Feed ratio Feeding amount mmol 2-3 461 1eq 9.2g 20 Ethanolamine 61 1eq 6.1g 100 Acetonitrile - - 80mL -

[0285] Operation process:

[0286] 2-3, ethanolamine, and acetonitrile were added to the reaction flask and stirred at room temperature for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.3 for the product. The reaction mixture was diluted with 200 mL of ethyl acetate and washed twice with 200 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 5.6 g of a colorless oil.

[0287] Step 5: Preparation of compound 2

[0288] Reaction formula:

[0289]

[0290] Material ratio:

[0291]

[0292]

[0293] Operation process:

[0294] Add 2-2, 2-4, potassium carbonate, potassium iodide, and acetonitrile to a reaction flask and heat to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) reveals an Rf value of 0.5 for the product. Dilute the reaction solution with 100 mL of ethyl acetate and wash once with 100 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.2 g of a colorless oil.

[0295] 1 H NMR(600MHz,Chloroform-d)δ5.08(qd,J=6.9,3.3Hz,1H),4.85(p,J=6.2Hz,1H),4.21(dd,J=11.8,3.3Hz,1H),4.01(dd,J=11.8,6.8Hz,1 H),3.69–3.59(m,2H),2.66(d,J=75.2Hz,4H),2.28(dt,J=14.1,7.5Hz,6H),1.65–1.45(m,16H),1.36–1.20(m,56H),0.91–0.84(m,12H).

[0296] MS (ES+) m / z): 852 (M).

[0297] Preparation Example 2 Preparation of Compound 3

[0298] Step 1: Preparation of 3-1

[0299] Reaction formula:

[0300]

[0301] Material ratio:

[0302] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1eq 5g 27 5-Bromopentanoic acid 181 1.1 eq 5.4g 30 Ferric chloride 162 0.025 eq 220mg 1.35 Pyridine 79 0.0125eq 53mg 0.68

[0303] Operation process:

[0304] 5-Bromopentanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a 250 mL reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 4:1) indicated completion of the reaction (product rf value 0.5). Purification by column chromatography yielded 6.4 g of a colorless oil.

[0305] Step 2: Preparation of 3-2

[0306] Reaction formula:

[0307]

[0308] Material ratio:

[0309]

[0310]

[0311] Operation process:

[0312] To the reaction flask, 3-1, octanoic acid, EDCI, DMAP, and dichloromethane were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 6.5 g of a colorless oil.

[0313] Step 3: Preparation of compound 3

[0314] Reaction formula:

[0315]

[0316] Material ratio:

[0317] Material Name Molecular weight Feed ratio Feeding amount mmol 3-2 491.55 1.1eq 4.2g 8.55 2-4 441.74 1 eq 3.4g 7.77 K2CO3 138 1.5eq 1.6g 11.6 KI 166 1.5eq 1.9g 11.6 Acetonitrile - - 40mL -

[0318] Operation process:

[0319] To the reaction flask were added 3-2, 2-4, K2CO3, KI, and acetonitrile, and the mixture was heated to 65°C and stirred for 16 h. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was filtered, concentrated, and purified by column chromatography to yield 3.6 g of a colorless oil.

[0320] 1 H NMR(600MHz,Chloroform-d)δ5.08(qd,J=7.3,3.2Hz,1H),4.86(p,J=6.2Hz,1H),4.27–4.21(m,1H),3.99(dd,J=11.8,7.0Hz,1H),3.68( d,J=6.3Hz,2H),2.80–2.59(m,6H),2.30(ddt,J=23.3,15.5,7.0Hz,6H),1.65–1.47(m,16H),1.35–1.21(m,54H),0.87(t,J=7.0Hz,12H).

[0321] MS (ES+) m / z): 852.7 (M+H) + .

[0322] Preparation Example 3 Preparation of Compound 5

[0323] Step 1: Preparation of 5-1

[0324] Reaction formula:

[0325]

[0326] Material ratio:

[0327] Material Name Molecular weight Feed ratio Feeding amount mmol Decyl epoxide 156 1eq 4.2g 27 8-Bromooctanoic acid 223 1.1 eq 6.7g 30 Ferric chloride 162 0.025 eq 220mg 1.35 Pyridine 79 0.0125eq 53mg 0.68

[0328] Operation process:

[0329] To a 250 mL reaction flask, add 8-bromooctanoic acid, decyl oxide, ferric chloride, and pyridine. Stir and react at room temperature for 16 hours. TLC (PE:EA = 4:1) indicates completion of the reaction (product rf value 0.5). Purify by column chromatography to yield 6.9 g of a colorless oil.

[0330] Step 2: Preparation of 5-2

[0331] Reaction formula:

[0332]

[0333] Material ratio:

[0334] Material Name Molecular weight Feed ratio Feeding amount mmol 5-1 379.38 1eq 6.6g 17.5 bitter 144 1.1 eq 2.8g 19.2 EDCI 192 2 eq 6.7g 35 DMAP 122 0.2 eq 420mg 3.5 dichloromethane - - 80mL -

[0335] Operation process:

[0336] To the reaction flask, 5-1, octanoic acid, EDCI, DMAP, and dichloromethane were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 6 g of a colorless oil.

[0337] Step 3: Preparation of compound 5

[0338] Reaction formula:

[0339]

[0340] Material ratio:

[0341] Material Name Molecular weight Feed ratio Feeding amount mmol 5-2 505.58 2.2eq 4.3g 8.55 Ethanolamine 61 1 eq 230mg 3.9 K2CO3 138 2 eq 1.1g 7.8 KI 166 2 eq 1.3g 7.8 Acetonitrile - - 40mL -

[0342] Operation process:

[0343] 5-2, ethanolamine, KCO, KI, and acetonitrile were added to the reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was filtered, concentrated, and purified by column chromatography to yield 2.5 g of a colorless oil.

[0344] 1 H NMR(600MHz,Chloroform-d)δ5.07(qd,J=6.7,3.2Hz,2H),4.21(dd,J=11.8,3.4Hz,2H),4.01(dd,J=11.8,6.8Hz,2H),2.67(t,J=5.6Hz,2H) ,2.54(t,J=7.8Hz,4H),2.29(t,J=7.3Hz,8H),1.64–1.52(m,12H),1.49(t,J=7.9Hz,4H),1.34–1.20(m,54H),0.87(td,J=7.0,2.0Hz,12H).

[0345] MS (ES+) m / z): 910.7 (M+H) + .

[0346] Preparation Example 4 Preparation of Compound 6

[0347] Step 1: Preparation of 6-1

[0348] Reaction formula:

[0349]

[0350] Material ratio:

[0351] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1eq 5g 27 6-Bromohexanoic acid 209 1.1eq 5.85g 30 Ferric chloride 162 0.025 eq 220mg 1.35 Pyridine 79 0.0125eq 53mg 0.68

[0352] Operation process:

[0353] To a 250 mL reaction flask, add 6-bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine. Stir and react at room temperature for 16 hours. TLC (PE:EA = 4:1) indicates completion of the reaction (product rf value 0.5). Purification by column chromatography yields 6.8 g of a colorless oil.

[0354] Step 2: Preparation of 6-2

[0355] Reaction formula:

[0356]

[0357] Material ratio:

[0358] Material Name Molecular weight Feed ratio Feeding amount mmol 6-1 379.38 1eq 6.6g 17.5 bitter 144 1.1 eq 2.8g 19.2 EDCI 192 2 eq 6.7g 35 DMAP 122 0.2 eq 420mg 3.5 dichloromethane - - 80mL -

[0359] Operation process:

[0360] To the reaction flask, 6-1, octanoic acid, EDCI, DMAP, and dichloromethane were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed twice with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 6.5 g of a colorless oil.

[0361] Step 3: Preparation of compound 6

[0362] Reaction formula:

[0363]

[0364] Material ratio:

[0365] Material Name Molecular weight Feed ratio Feeding amount mmol 6-2 505.58 2.2eq 4.3g 8.55 Ethanolamine 61 1 eq 230mg 3.9 K2CO3 138 2 eq 1.1g 7.8 KI 166 2 eq 1.3g 7.8 Acetonitrile - - 40mL -

[0366] Operation process:

[0367] 6-2, ethanolamine, KCO, KI, and acetonitrile were added to the reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was filtered, concentrated, and purified by column chromatography to yield 2.8 g of a colorless oil.

[0368] 1 H NMR(600MHz,Chloroform-d)δ5.10–5.05(m,2H),4.22(dd,J=11.8,3.3Hz,2H),4.01(dd,J=11.8,6.8Hz,2H),3.61(d,J= 5.1Hz,2H),2.73–2.49(m,6H),2.30(td,J=7.5,5.1Hz,8H),1.65–1.50(m,16H),1.33–1.23(m,52H),0.91–0.85(m,12H).

[0369] MS (ES+) m / z): 910.7 (M+H) + .

[0370] Preparation Example 5 Preparation of Compound 8

[0371] Step 1: Preparation of 8-1

[0372] Reaction formula:

[0373]

[0374] Material ratio:

[0375] Material Name Molecular weight Feed ratio Feeding amount mmol 1-nonanol 144 1eq 7.2g 50 6-Bromohexanoic acid 195 1.1eq 10.7g 55 EDCI 192 2eq 19.2g 100 DMAP 122 0.2eq 1.2g 10 dichloromethane - - 200mL -

[0376] Operation process:

[0377] 1-Nonanol, 6-bromohexanoic acid, EDCI, DMAP, and dichloromethane were added to the reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed twice with 200 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 13 g of a colorless oil.

[0378] Step 2: Preparation of 8-2

[0379] Reaction formula:

[0380]

[0381] Material ratio:

[0382] Material Name Molecular weight Feed ratio Feeding amount mmol 8-1 321.3 1eq 13g 40.5 Ethanolamine 61 10eq 24.7g 405 Acetonitrile - - 200mL -

[0383] Operation process:

[0384] 8-1, ethanolamine, and acetonitrile were added to the reaction flask and heated to 30°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.1). The reaction solution was concentrated and diluted with 200 mL of ethyl acetate and washed twice with 200 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of a colorless oil.

[0385] Step 3: Preparation of compound 8

[0386] Reaction formula:

[0387]

[0388] Material ratio:

[0389] Material Name Molecular weight Feed ratio Feeding amount mmol 2-2 491.55 1.1eq 4.2g 8.55 8-2 301.47 1eq 2.34g 7.77 K2CO3 138 1.5eq 1.6g 11.6 KI 166 1.5eq 1.9g 11.6 Acetonitrile - - 40mL -

[0390] Operation process:

[0391] 2-2, 8-2, K2CO3, KI, and acetonitrile were added to the reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was filtered, concentrated, and purified by column chromatography to yield 2.6 g of a colorless oil.

[0392] 1H NMR(600MHz,Chloroform-d)δ5.07(qd,J=6.9,3.3Hz,1H),4.21(dd,J=11.8,3.3Hz,1H),4.06–3.99(m,3H),3.53(t,J=5.3Hz,2H),2.58(t,J=5.3Hz ,2H),2.49–2.43(m,4H),2.29(t,J=7.5Hz,6H),1.66–1.52(m,10H),1.45 (dp,J=14.3,7.5Hz,4H),1.35–1.22(m,38H),0.87(td,J=7.0,2.2Hz,9H).

[0393] MS (ES+) m / z): 712.6 (M+H) + .

[0394] Preparation Example 6 Preparation of Compound 9

[0395] Step 1: Preparation of compound 9

[0396] Reaction formula:

[0397]

[0398] Material ratio:

[0399] Material Name Molecular weight Feed ratio Feeding amount mmol 3-2 491.55 1.1eq 4.2g 8.55 8-2 301.47 1eq 2.34g 7.77 <![CDATA[K2CO3]]> 138 1.5eq 1.6g 11.6 KI 166 1.5eq 1.9g 11.6 Acetonitrile - - 40mL -

[0400] Operation process:

[0401] 3-2, 8-2, K2CO3, KI, and acetonitrile were added to the reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was filtered, concentrated, and purified by column chromatography to yield 2.4 g of a colorless oil.

[0402] 1 H NMR(600MHz,Chloroform-d)δ5.08(qd,J=7.0,3.2Hz,1H),4.22(dd,J=11.8,3.3Hz,1H),4.07–3.99(m,3H),3.53(t,J=5.3Hz,2H),2.57(t, J=5.4Hz,2H),2.46(q,J=8.3Hz,4H),2.33–2.27(m,6H),1.65–1.53(m,10H),1.50–1.43(m,4H),1.34–1.22(m,38H),0.87(t,J=6.8Hz,9H).

[0403] MS (ES+) m / z): 712.6 (M+H) + .

[0404] Preparation Example 7 Preparation of Compound 27

[0405] Step 1: Synthesis of 2-hydroxydodecane 5-bromopentyl ester

[0406] Reaction formula:

[0407]

[0408] Material ratio:

[0409] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1.0 eq 10.0g 54.3mmol 5-Bromopentanoic acid 181 1.025eq 10.3g 57.0mmol Ferric chloride 162 0.025 eq 220mg 1.36mmol Pyridine 79 0.028 eq 110mg 1.52mmol

[0410] Operation process:

[0411] To the reaction flask were added 1,2-epoxydodecane (10.0 g, 54.3 mmol), 5-bromovaleric acid (10.3 g, 57.0 mmol), ferric chloride (220 mg, 1.36 mmol), and pyridine (120 mg, 1.52 mmol) in sequence. The mixture was then reacted at 20°C under nitrogen for 12 hours. The reaction was monitored by TLC, and the reaction solution was concentrated and purified by column chromatography to obtain a colorless liquid, 2-hydroxydodecane 5-bromopentyl ester (13.1 g, 66.1% yield).

[0412] Step 2: Synthesis of 1-((5-bromovaleryl)oxy)dodec-2-ylhexyl ester

[0413] Reaction formula:

[0414]

[0415] Material ratio:

[0416]

[0417] Operation process:

[0418] 2-Hydroxydodecane 5-bromopentyl ester (5.00 g, 13.69 mmol) was dissolved in dichloromethane (50 mL). Hexanoic acid (2.07 g, 17.8 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.94 g, 20.5 mmol), and 4-dimethylaminopyridine (167 mg, 1.37 mmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-((5-bromovaleroyl)oxy)dodec-2-ylhexyl ester (4.00 g, 63.1% yield) as a colorless liquid.

[0419] Step 3: Synthesis of 1-({5-[(5-{[2-(hexanoyloxy)dodecyl]oxy}-5-oxypentyl)(2-hydroxyethyl)amino]pentanoyl}oxy)dodec-2-ylhexyl ester

[0420] Reaction formula:

[0421]

[0422] Material ratio:

[0423]

[0424] Operation process:

[0425] 1-((5-bromovaleroyl)oxy)dodec-2-ylhexyl ester (2.00 g, 4.32 mmol) was dissolved in acetonitrile (10 mL), and ethanolamine (121 mg, 1.98 mmol), potassium carbonate (2.09 g, 15.1 mmol), potassium iodide (860 mg, 5.18 mmol), and tetrahydrofuran (10 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({5-[(5-{[2-(hexanoyloxy)dodecyl]oxy}-5-oxyidenepentyl)(2-hydroxyethyl)amino]pentanoyl}oxy)dodec-2-ylhexyl ester (700 mg, 19.6% yield) as a pale yellow liquid.

[0426] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.10 (dq, J1 = 3.2, J2 = 6.6Hz, 2H), 4.26 (dd, J1 = 3.2, J2 = 11.8Hz, 2H), 4.00 (dd, J1 = 7 .0,J2=11.8Hz,2H),3.85-3.77(m,1H),3.71(brd,J=2.8Hz,2H),3.58-3.52(m,1H),3.38(t,J=5.6Hz,1H),2.70(br s,5H),2.42(t,J=6.2Hz,1H),2.37-2.29(m,8H),1.86-1.81(m,1H),1.63(br t,J=7.0Hz,10H),1.56(br d,J=5.8Hz,2H),1.34-1.24(m,40H),0.93-0.86(m,12H)ppm.

[0427] LCMS:RT=1.985,m / z 826.6[M+H] + .

[0428] Preparation Example 8 Preparation of Compound 24

[0429] Step 1: Synthesis of heptadecan-9-yl 8-bromooctyl ester

[0430] Reaction formula:

[0431]

[0432] Material ratio:

[0433]

[0434] Operation process:

[0435] Dissolve heptadecan-9-ol (6.00 g, 23.4 mmol) in dichloromethane (60 mL). 8-Bromooctanoic acid (6.79 g, 30.4 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (6.73 g, 35.1 mmol), and 4-dimethylaminopyridine (286 mg, 2.34 mmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield colorless heptadecan-9-yl 8-bromooctyl ester (8.40 g, 77.8% yield).

[0436] Step 2: Synthesis of heptadecan-9-yl 8-((4-hydroxybutyl)amino)octyl ester

[0437] Reaction formula:

[0438]

[0439] Material ratio:

[0440]

[0441]

[0442] Operation process:

[0443] Dissolve heptadecan-9-yl 8-bromooctyl ester (1.00 g, 2.17 mmol) in acetonitrile (10 mL), then add 4-amino-1-butanol (1.93 g, 21.7 mmol). The mixture is reacted at 20°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated, washed with water, and extracted. Column chromatography of the crude product affords heptadecan-9-yl 8-((4-hydroxybutyl)amino)octyl ester (1.00 g, 98.3% yield) as a colorless liquid.

[0444] Step 3: Synthesis of heptadecane-9-yl 8-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(4-hydroxybutyl)amino]octyl ester

[0445] Reaction formula:

[0446]

[0447] Material ratio:

[0448]

[0449] Operation process:

[0450] Dissolve heptadecan-9-yl 8-[(4-hydroxybutyl)amino]octyl ester (600 mg, 1.28 mmol) in acetonitrile (6.00 mL). Add 2-(hexanoyloxy)dodecyl 6-bromohexyl ester (792 mg, 1.66 mmol), potassium carbonate (617 mg, 4.47 mmol), potassium iodide (254 mg, 1.53 mmol), and tetrahydrofuran (6.00 mL) in this order. The mixture is reacted at 80°C under nitrogen for 12 hours. The reaction is monitored by TLC, filtered, and concentrated. Purification by column chromatography affords heptadecan-9-yl 8-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(4-hydroxybutyl)amino]octyl ester (500 mg, 43.3% yield), a colorless liquid.

[0451] 1 H NMR (400MHz, CHLOROFORM-d) δ = 6.67-6.14 (m, 1H), 5.09 (dq, J1 = 3.4, J2 = 6.6Hz, 1H), 4.87 (t, J = 6.2Hz, 1H), 4.23 (dd, J1 = 3.4, J2 = 11.8Hz, 1H), 4.03 (d d, J1=6.8, J2=11.8Hz,1H),3.64-3.49(m,2H),2.50-2.36(m,6H),2.35-2. 22(m,6H),1.66-1.42(m,22H),1.34-1.23(m,50H),0.95-0.78(m,12H)ppm.

[0452] LCMS:RT=2.124,m / z 866.7[M+H] + .

[0453] Preparation Example 9 Preparation of Compound 23

[0454] Step 1: Synthesis of 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(3-hydroxypropyl)amino]hexyl ester

[0455] Reaction formula:

[0456]

[0457] Material ratio:

[0458]

[0459] Operation process:

[0460] 2-(Hexanoyloxy)dodecyl 6-bromohexyl ester (2.00 g, 4.19 mmol) was dissolved in acetonitrile (10 mL), and 3-aminopropanol (142 mg, 1.88 mmol), potassium carbonate (2.03 g, 14.7 mmol), potassium iodide (834 mg, 5.03 mmol), and tetrahydrofuran (10 mL) were added in sequence. The mixture was reacted at 80° C. under nitrogen for 8 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(3-hydroxypropyl)amino]hexyl ester (800 mg, 22.0% yield) as a pale yellow liquid.

[0461] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.13-5.05 (m, 2H), 4.23 (dd, J1 = 2.8, J2 = 11.8Hz, 2H), 4.03 (dd, J1 = 6.8, J2 = 11.8Hz, 2H), 3.79 (br t, J = 4.8Hz, 2H), 2.67 (br s,2H),2.51-2.40(m,4H),2.31(dt,J1=3.6,J2=7.2Hz,8H),1.72-1.47(m,20H),1.35-1.25(m,42H),0.89(q,J=6.8Hz,12H)ppm.

[0462] LCMS:RT=5.629,m / z 868.5[M+H] + .

[0463] Preparation Example 10 Preparation of Compound 25

[0464] Step 1: Synthesis of 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(4-hydroxybutyl)amino]hexyl ester

[0465] Reaction formula:

[0466]

[0467] Material ratio:

[0468]

[0469] Operation process:

[0470] 2-(Hexanoyloxy) dodecyl 6-bromohexyl ester (2.00 g, 4.19 mmol) was dissolved in acetonitrile (10 mL), and 4-aminobutanol (168 mg, 1.88 mmol), potassium carbonate (2.03 g, 14.7 mmol), potassium iodide (834 mg, 5.03 mmol), and tetrahydrofuran (10 mL) were added in sequence. The mixture was reacted at 80° C. for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a light yellow liquid of 2-(hexanoyloxy) dodecyl 6-[(6-{[2-(hexanoyloxy) dodecyl] oxy}-6-oxyylidenehexyl)(4-hydroxybutyl) amino] hexyl ester (800 mg, 21.7% yield).

[0471] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.14-5.04 (m, 2H), 4.23 (dd, J1 = 3.2, J2 = 11.8Hz, 2H), 4.02 (dd J1 = 6.8, J2 = 11.8Hz, 2H), 3.58 (br s, 2H), 2.54 (br s,6H),2.31(dt,J1=4.2,J2=7.2Hz,8H),1.73-1.52(m,20H),1.36-1.23(m,44H),0.89(q,J=6.8Hz,12H)ppm.

[0472] LCMS:RT=5.781,m / z 882.5[M+H] +

[0473] Preparation Example 11 Preparation of Compound 26

[0474] Step 1: Synthesis of 2-hydroxydodecane 6-bromohexyl ester

[0475] Reaction formula:

[0476]

[0477] Material ratio:

[0478] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1.0 eq 10.0g 54.3mmol 6-Bromohexanoic acid 181 1.1eq 11.6g 59.7mmol Ferric chloride 162 0.025 eq 220mg 1.36mmol Pyridine 79 0.028 eq 120mg 1.52mmol

[0479] Operation process:

[0480] 1,2-Epoxydodecane (10.0 g, 54.3 mmol), 6-bromohexanoic acid (11.6 g, 59.7 mmol), ferric chloride (220 mg, 1.36 mmol), and pyridine (120 mg, 1.52 mmol) were added sequentially to a reaction flask and reacted at 20°C under nitrogen for 12 hours. The reaction was monitored by TLC, and the reaction solution was concentrated to remove pyridine. Purification by column chromatography afforded a colorless liquid, 2-hydroxydodecyl 6-bromohexyl ester (14.5 g, 70.5% yield).

[0481] Step 2: Synthesis of 2-(hexanoyloxy)dodecane 6-bromohexyl ester

[0482] Reaction formula:

[0483]

[0484] Material ratio:

[0485]

[0486] Operation process:

[0487] 2-Hydroxydodecane 6-bromohexyl ester (11 g, 29.0 mmol) was dissolved in dichloromethane (100 mL). Hexanoic acid (4.38 g, 37.7 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (8.34 g, 43.5 mmol), and 4-dimethylaminopyridine (354 mg, 2.90 mmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-(hexanoyloxy)dodecane 6-bromohexyl ester (12.5 g, 90.3% yield) as a colorless liquid.

[0488] Step 3: Synthesis of 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester

[0489] Reaction formula:

[0490]

[0491] Material ratio:

[0492]

[0493] Operation process:

[0494] 2-(Hexanoyloxy)dodecyl 6-bromohexyl ester (2.50 g, 5.24 mmol) was dissolved in acetonitrile (10 mL), and ethanolamine (150 mg, 2.46 mmol), potassium carbonate (1.19 g, 8.59 mmol), potassium iodide (938 mg, 5.65 mmol), and tetrahydrofuran (10 mL) were added in sequence. The mixture was reacted at 80° C. under nitrogen for 6 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a light yellow liquid 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester (1.10 g, 52.4% yield).

[0495] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (br dd, J1 = 3.0, J2 = 6.3Hz, 2H), 4.24 (dd, J1 = 3.2, J2 = 11.8Hz, 2H), 4.02 (dd, J1 = 6.8, J2 = 11.8Hz, 2H), 3.62 (br s,2H),2.69(br s,2H),2.58(br s,4H),2.31(dt,J1=3.8,J2=7.4Hz,8H),1.68-1.52(m,16H),1.36-1.24(m,44H),0.89(q,J=7.0Hz,12H)ppm.

[0496] LCMS:RT=2.013,m / z 854.6[M+H] +

[0497] Preparation Example 12 Preparation of Compound 28

[0498] Step 1: Synthesis of heptadecan-9-yl 8-((2-hydroxyethyl)amino)octyl ester

[0499] Reaction formula:

[0500]

[0501] Material ratio:

[0502] Material Name Molecular weight Feed ratio Feeding amount mmol Heptadecan-9-yl 8-bromooctyl ester 460 1.0 eq 6.00g 13.0mmol Ethanolamine 61 10eq 7.94g 130mmol Acetonitrile - - 60mL - Tetrahydrofuran - - 20mL -

[0503] Operation process:

[0504] Dissolve heptadecan-9-yl 8-bromooctyl ester (6.00 g, 13.0 mmol) in acetonitrile (60 mL), then add ethanolamine (7.94 g, 130 mmol) and tetrahydrofuran (20 mL). The mixture is reacted at 20°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated, washed with water, and extracted. Column chromatography of the crude product affords heptadecan-9-yl 8-((2-hydroxyethyl)amino)octyl ester (3.40 g, 59.2% yield) as a colorless liquid.

[0505] Step 2: Synthesis of heptadec-9-yl 8-((6-((2-(hexanoyloxy)dodecyl)oxy)-6-oxyylidenehexyl)(2-hydroxyethyl)amino)octyl ester

[0506] Reaction formula:

[0507]

[0508] Material ratio:

[0509]

[0510]

[0511] Operation process:

[0512] Dissolve heptadecan-9-yl 8-((2-hydroxyethyl)amino)octyl ester (1.50 g, 3.40 mmol) in acetonitrile (8 mL), then add 2-(hexanoyloxy)dodecane 6-bromohexyl ester (1.78 g, 3.74 mmol), potassium carbonate (1.64 g, 11.9 mmol), potassium iodide (676 mg, 4.07 mmol), and tetrahydrofuran (8 mL). The mixture is reacted at 80°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC, filtered, and concentrated. Purification by column chromatography affords a pale yellow liquid heptadecan-9-yl 8-((6-((2-(hexanoyloxy)dodecyl)oxy)-6-oxyylidenehexyl)(2-hydroxyethyl)amino)octyl ester (1.22 g, 42.8% yield).

[0513] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.4, J2 = 6.6Hz, 1H), 4.87 (quin, J = 6.2Hz, 1H),4.23(dd,J1=3.4,J2=11.8Hz,1H),4.03(dd,J1=6.8,J2=11.8Hz,1H),3.52(t,J=5 .2Hz,2H),3.21-2.75(m,1H),2.57(t,J=5.2Hz,2H),2.48-2.39(m,4H),2.36-2.24(m, 6H),1.68-1.55(m,8H),1.54-1.42(m,8H),1.35-1.22(m,52H),1.00-0.79(m,12H)ppm.

[0514] LCMS:RT=5.920,m / z 838.6[M+H] + .

[0515] Preparation Example 13 Preparation of Compound 34

[0516] Step 1: Synthesis of 1-((6-bromohexanoyl)oxy)dodec-2-ylheptyl ester

[0517] Reaction formula:

[0518]

[0519] Material ratio:

[0520]

[0521] Operation process:

[0522] 6-Bromohexyl 2-hydroxydodecane (3.50 g, 9.23 mmol) was dissolved in dichloromethane (35 mL). Heptanoic acid (1.56 g, 12.0 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.65 g, 13.8 mmol), and 4-dimethylaminopyridine (113 mg, 923 μmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was concentrated, washed with water, and extracted. The crude product was purified by column chromatography to obtain 1-((6-bromohexanoyl)oxy)dodec-2-ylheptyl ester (3.60 g, 79.4% yield), a colorless liquid.

[0523] Step 2: Synthesis of heptadec-9-yl 8-((6-((2-(heptanoyloxy)dodecyl)oxy)-6-oxyylidenehexyl)(2-hydroxyethyl)amino)octyl ester

[0524] Reaction formula:

[0525]

[0526] Material ratio:

[0527]

[0528] Operation process:

[0529] 1-((6-bromohexanoyl)oxy)dodec-2-ylheptyl ester (1.50 g, 3.40 mmol) was dissolved in acetonitrile (8 mL), and heptadecan-9-yl 8-((2-hydroxyethyl)amino)octyl ester (1.84 g, 3.74 mmol), potassium carbonate (1.64 g, 11.9 mmol), potassium iodide (676 mg, 4.07 mmol), and tetrahydrofuran (8 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadecan-9-yl 8-((6-((2-(heptanoyloxy)dodecyl)oxy)-6-oxyylidenehexyl)(2-hydroxyethyl)amino)octyl ester (900 mg, 31.1% yield) as a pale yellow liquid.

[0530] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.13-5.05 (m, 1H), 4.86 (quin, J = 6.2Hz, 1H), 4.23 (dd, J1 = 3.2, J2 = 11.8Hz, 1H), 4.02 (dd, J1 = 6.8, J2 = 11.8Hz, 1H), 3.56 (br t,J=5.2Hz,2H),2.61(br t,J=5.0Hz,2H),2.53-2.44(m,4H),2.35-2.25(m,6H),1.66-1.45(m,16H),1.36-1.23(m,54H),0.97-0.79(m,12H)ppm.

[0531] LCMS:RT=7.550,m / z 852.6[M+H] + .

[0532] Preparation Example 14 Preparation of Compound 29

[0533] Step 1: Synthesis of heptadec-9-yl 8-((5-((2-(hexanoyloxy)dodecyl)oxy)-5-oxypentylidene)(2-hydroxyethyl)amino)octyl ester

[0534] Reaction formula:

[0535]

[0536] Material ratio:

[0537]

[0538] Operation process:

[0539] Dissolve heptadecan-9-yl 8-((2-hydroxyethyl)amino)octyl ester (1.50 g, 3.40 mmol) in acetonitrile (8 mL), then add 1-((5-bromovaleryl)oxy)dodec-2-ylhexyl ester (1.73 g, 3.74 mmol), potassium carbonate (1.68 g, 11.9 mmol), potassium iodide (676 mg, 4.07 mmol), and tetrahydrofuran (8 mL). The mixture is reacted at 80°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC, filtered, and concentrated. Purification by column chromatography affords a pale yellow liquid heptadecan-9-yl 8-((5-((2-(hexanoyloxy)dodecyl)oxy)-5-oxypentylene)(2-hydroxyethyl)amino)octyl ester (1.66 g, 59.3% yield).

[0540] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.10 (dq, J1 = 3.2, J2 = 6.6Hz, 1H), 4.87 (t, J = 6.2Hz, 1H),4.25(dd,J1=3.2,J2=11.8Hz,1H),4.02(dd,J1=6.8,J2=11.8Hz,1H),3.59(br t,J=4.8Hz,2H),2.65(br s,2H),2.57-2.48(m,4H),2.36-2.26(m,6H),1.68-1.44(m,18H),1.34-1.25(m,48H),0.93-0.85(m,12H)ppm.

[0541] LCMS:RT=2.184,m / z 824.7[M+H] + .

[0542] Preparation Example 15 Preparation of Compound 4

[0543] Step 1: Synthesis of 2-hydroxyundecyl 7-bromoheptyl ester

[0544] Reaction formula:

[0545]

[0546] Material ratio:

[0547] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Nonyloxadiazole 170 1.0 eq 4.00g 23.4mmol 7-Bromoheptanoic acid 208 1.05eq 5.16g 24.6mmol Ferric chloride 162 0.025eq 95.2mg 0.587mmol Pyridine 79 0.025eq 46.4mg 0.587mmol

[0548] Operation process:

[0549] To a reaction flask, 2-nonyloxirane (4.00 g, 23.4 mmol), 7-bromoheptanoic acid (5.16 g, 24.6 mmol), ferric chloride (95.2 mg, 587 μmol), and pyridine (46.4 mg, 587 μmol) were added sequentially. The mixture was then reacted at 30°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction solution was concentrated and purified by column chromatography to yield 2-hydroxyundecyl 7-bromoheptyl ester (6.30 g, 70.3% yield) as a colorless liquid.

[0550] Step 2: Synthesis of 1-[(7-bromoheptanoyl)oxy]undec-2-yloctyl ester

[0551] Reaction formula:

[0552]

[0553] Material ratio:

[0554]

[0555] Operation process:

[0556] 2-Hydroxyundecyl 7-bromoheptyl ester (3.00 g, 7.91 mmol) was dissolved in dichloromethane (3.00 mL). Octanoic acid (1.37 g, 9.49 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27 g, 11.8 mmol), and 4-dimethylaminopyridine (96.6 mg, 790 μmol) were added sequentially. The mixture was reacted under nitrogen for 12 hours at 25°C. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-[(7-bromoheptyl)oxy]undec-2-yloctyl ester (2.95 g, 73.7% yield) as a colorless liquid.

[0557] Step 3: Synthesis of 1-({7-[(2-hydroxyethyl)(7-{[2-(octanoyloxy)undecyl]oxy}-7-oxyylideneheptyl)amino]heptanoyl}oxy)undec-2-yloctyl ester

[0558] Reaction formula:

[0559]

[0560] Material ratio:

[0561] Material Name Molecular weight Feed ratio Feeding amount mmol 1-[(7-bromoheptanoyl)oxy]undec-2-yloctyl ester 504 1.0 eq 1.30g 2.57mmol Ethanolamine 61 0.45eq 70.6mg 1.16mmol potassium carbonate 138 3.5eq 1.24g 9.00mmol Potassium iodide 166 1.2 eq 512mg 3.09mmol Acetonitrile - - 10.0mL - Tetrahydrofuran - - 3.00mL -

[0562] Operation process:

[0563] 1-[(7-bromoheptanoyl)oxy]undec-2-yloctyl ester (1.30 g, 2.57 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (70.6 mg, 1.16 mmol), potassium carbonate (1.24 g, 9.00 mmol), potassium iodide (512 mg, 3.09 mmol), and tetrahydrofuran (3.00 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({7-[(2-hydroxyethyl)(7-{[2-(octanoyloxy)undecyl]oxy}-7-oxyylideneheptyl)amino]heptanoyl}oxy)undec-2-yloctyl ester (546 mg, 21.8% yield) as a colorless liquid.

[0564] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.4, J2 = 6.6Hz, 2H), 4.22 (dd, J1 = 3.4, J2=11.8Hz,2H),4.03(dd,J1=6.8,J2=11.8Hz,2H),3.56(t,J=5.4Hz,2H),2.62(br t,J=5.2Hz,2H),2.55-2.42(m,4H),2.30(t,J=7.6Hz,8H),1.70-1.54(m,12H ),1.47(td,J=7.4,14.2Hz,4H),1.35-1.18(m,52H),0.95-0.77(m,12H)ppm.

[0565] LCMS:RT=2.130,m / z 910.6[M+H] + .

[0566] Preparation Example 16 Preparation of Compound 1

[0567] Step 1: Synthesis of 2-nonyloxadiazole

[0568] Reaction formula:

[0569]

[0570] Material ratio:

[0571] Material Name Molecular weight Feed ratio Feeding amount mmol Undec-1-ene 154 1.00 eq 10.0g 64.8mmol 3-Chlorobenzoperoxyacid 172 1.20eq 15.7g 77.7mmol dichloromethane - - 140mL -

[0572] Operation process:

[0573] Undec-1-ene (10.0 g, 64.8 mmol) and dichloromethane (140 mL) were added sequentially to a reaction flask. 3-Chlorobenzoperoxyacid (15.7 g, 77.7 mmol) was slowly added at 0°C. Finally, the mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC, and the reaction mixture was quenched, extracted, and purified by column chromatography to afford 2-nonyloxadiazole (11.0 g, 99.6% yield) as a colorless liquid.

[0574] Step 2: Synthesis of 2-hydroxyundecyl 6-bromohexyl ester

[0575] Reaction formula:

[0576]

[0577] Material ratio:

[0578] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Nonyloxadiazole 170 1.00 eq 4.00g 23.4mmol 6-Bromohexanoic acid 193 1.05eq 4.81g 24.6mmol Ferric chloride 162 0.025eq 95.2mg 0.587mmol Pyridine 79 0.025eq 46.4mg 0.587mmol

[0579] Operation process:

[0580] To a reaction flask, 2-nonyloxirane (4.00 g, 23.4 mmol), 6-bromohexanoic acid (4.81 g, 24.6 mmol), ferric chloride (95.2 mg, 587 μmol), and pyridine (46.4 mg, 587 μmol) were added sequentially. The mixture was reacted at 30°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction solution was concentrated and purified by column chromatography to yield 2-hydroxyundecyl 6-bromohexyl ester (6.50 g, 75.5% yield), a colorless liquid.

[0581] Step 3: Synthesis of 1-[(6-bromohexanoyl)oxy]undec-2-yloctyl ester

[0582] Reaction formula:

[0583]

[0584] Material ratio:

[0585]

[0586] Operation process:

[0587] Dissolve 2-hydroxyundecyl 6-bromohexyl ester (3.00 g, 8.21 mmol) in dichloromethane (30.0 mL). Add n-octanoic acid (1.42 g, 9.85 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.36 g, 12.3 mmol), and 4-dimethylaminopyridine (100 mg, 821 μmol) sequentially. The mixture is reacted at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC, and the reaction solution is concentrated. Column chromatography of the crude product affords 1-[(6-bromohexanoyl)oxy]undec-2-yloctyl ester (2.74 g, 67.8% yield) as a colorless liquid.

[0588] Step 4: Synthesis of 1-({6-[(2-hydroxyethyl)amino]hexanoyl}oxy)undec-2-yloctyl ester

[0589] Reaction formula:

[0590]

[0591] Material ratio:

[0592] Material Name Molecular weight Feed ratio Feeding amount mmol 1-[(6-bromohexanoyl)oxy]undec-2-yloctyl ester 490 1.00 eq 1.00g 2.03mmol Ethanolamine 61 10.0eq 1.24g 20.3mmol Acetonitrile - - 22.0mL -

[0593] Operation process:

[0594] Dissolve 1-[(6-bromohexanoyl)oxy]undec-2-yloctyl ester (1.00 g, 2.03 mmol) in acetonitrile (22.0 mL), followed by the addition of ethanolamine (1.24 g, 20.3 mmol). The mixture was reacted at 30°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, followed by filtration and concentration. Purification by column chromatography afforded 1-({6-[(2-hydroxyethyl)amino]hexanoyl}oxy)undec-2-yloctyl ester (840 mg, 93.0% yield) as a colorless liquid.

[0595] Step 5: Synthesis of heptadec-9-yl 8-[(2-hydroxyethyl)(6-{[2-(octanoyloxy)undecyl]oxy}-6-oxyylidenehexyl)amino]octyl ester

[0596] Reaction formula:

[0597]

[0598] Material ratio:

[0599]

[0600] Operation process:

[0601] 2-(Octyloxy)undecyl 5-[(2-hydroxyethyl)amino]pentyl ester (740 mg, 1.57 mmol) was dissolved in acetonitrile (7.00 mL). Heptadec-9-yl 8-bromooctyl ester (868 mg, 1.88 mmol), potassium carbonate (758 mg, 5.49 mmol), potassium iodide (312 mg, 1.88 mmol), and tetrahydrofuran (7.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadec-9-yl 8-[(2-hydroxyethyl)(6-{[2-(octanoyloxy)undecyl]oxy}-6-oxyylidenehexyl)amino]octyl ester (700 mg, 19.6% yield) as a colorless liquid.

[0602] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.16-5.01(m,1H),4.87(t,J=6.2Hz,1H),4.23(dd,J1= 3.4, J2=11.8Hz, 1H), 4.02(dd, J1=6.8, J2=11.8Hz, 1H), 3.58(t, J=5.2Hz, 2H), 2.63(br t,J=5.2Hz,2H),2.57-2.43(m,4H),2.34-2.23(m,6H),1.65-1.42(m,16H),1.34-1.16(m,54H),0.88(t,J=6.8Hz,12H)ppm.

[0603] LCMS:RT=2.187,m / z 852.7[M+H] + .

[0604] Preparation Example 17 Preparation of Compound 7

[0605] Step 1: Synthesis of 2-heptyloxadiazole

[0606] Reaction formula:

[0607]

[0608] Material ratio:

[0609] Material Name Molecular weight Feed ratio Feeding amount mmol Non-1-ene 126 1.00 eq 4.00g 31.6mmol 3-Chlorobenzoperoxyacid 172 1.20eq 7.72g 38.0mmol dichloromethane 51.0mL

[0610] Operation process:

[0611] Undec-1-ene (4.00 g, 31.6 mmol) and dichloromethane (51.0 mL) were added sequentially to a reaction flask. 3-Chlorobenzoperoxyacid (7.72 g, 38.0 mmol) was then slowly added at 0°C. The mixture was then allowed to react at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC, and the resulting solution was quenched, extracted, and purified by column chromatography to afford 2-heptyloxadiazole (3.95 g, 87.6% yield) as a colorless liquid.

[0612] Step 2: Synthesis of nonyl 6-bromohexyl ester,

[0613] Reaction formula:

[0614]

[0615] Material ratio:

[0616]

[0617] Operation process:

[0618] Dissolve nonan-1-ol (4 g, 27.7 mmol) in dichloromethane (40.0 mL). 6-bromohexanoic acid (6.49 g, 33.2 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.97 g, 41.5 mmol), and 4-dimethylaminopyridine (338 mg, 2.77 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 8.50 g of nonyl 6-bromohexyl ester (95.4% yield), a colorless liquid.

[0619] Step 3: Synthesis of 2-hydroxynonyl 8-bromooctyl ester

[0620] Reaction formula:

[0621]

[0622] Material ratio:

[0623] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Heptyloxadiazole 142 1.00 eq 3.34g 23.4mmol 8-Bromooctanoic acid 222 1.05eq 5.50g 24.6mmol Ferric chloride 162 0.025eq 95.2mg 0.587mmol Pyridine 79 0.028eq 52.0mg 0.657mmol

[0624] Operation process:

[0625] To a reaction flask, 2-heptyloxirane (3.34 g, 23.4 mmol), 8-bromooctanoic acid (5.50 g, 24.6 mmol), ferric chloride (95.2 mg, 587 μmol), and pyridine (52.0 mg, 657 μmol) were added sequentially. The mixture was reacted at 30°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction solution was concentrated and purified by column chromatography to yield 2-hydroxynonyl 8-bromooctyl ester (6.50 g, 75.7% yield), a colorless liquid.

[0626] Step 4: Synthesis of 2-(octanoyloxy)nonyl 8-bromooctyl ester

[0627] Reaction formula:

[0628]

[0629] Material ratio:

[0630]

[0631]

[0632] Operation process:

[0633] Dissolve 2-hydroxynonyl 8-bromooctyl ester (3.00 g, 8.21 mmol) in dichloromethane (30.0 mL). Add n-octanoic acid (1.42 g, 9.85 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.36 g, 12.3 mmol), and 4-dimethylaminopyridine (100 mg, 821 μmol) sequentially. Incubate at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. Column chromatography yields the crude product as a colorless liquid, 2-(octanoyloxy)nonyl 8-bromooctyl ester (3.00 g, 74.3% yield).

[0634] Step 5: Synthesis of 2-(octanoyloxy)nonyl 8-[(2-hydroxyethyl)amino]octyl ester

[0635] Reaction formula:

[0636]

[0637] Material ratio:

[0638] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Octanoyloxy)nonyl 8-bromooctyl ester 490 1.00 eq 1.20g 2.44mmol Ethanolamine 61 10.0eq 1.49g 24.4mmol Acetonitrile - - 22.0mL -

[0639] Operation process:

[0640] 2-(Octanoyloxy)nonyl 8-bromooctyl ester (1.20 g, 2.44 mmol) was dissolved in acetonitrile (22.0 mL), followed by the addition of ethanolamine (1.49 g, 24.4 mmol). The mixture was reacted at 30°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography afforded a colorless liquid, 2-(Octanoyloxy)nonyl 8-[(2-hydroxyethyl)amino]octyl ester (800 mg, 69.4% yield).

[0641] Step 6: Synthesis of 2-(octanoyloxy)nonyl 8-[(2-hydroxyethyl)[6-(nonyloxy)-6-oxyylidenehexyl]amino]octyl ester

[0642] Reaction formula:

[0643]

[0644] Material ratio:

[0645]

[0646]

[0647] Operation process:

[0648] 2-(Octanoyloxy)nonyl 8-[(2-hydroxyethyl)amino]octyl ester (700 mg, 1.48 mmol) was dissolved in acetonitrile (10.0 mL). Nonyl 6-bromohexyl ester (572 mg, 1.78 mmol), potassium carbonate (717 mg, 5.19 mmol), potassium iodide (295 mg, 1.78 mmol), and tetrahydrofuran (10.0 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(Octanoyloxy)nonyl 8-[(2-hydroxyethyl)[6-(nonyloxy)-6-oxyylidenehexyl]amino]octyl ester (780 mg, 69.2% yield) as a colorless liquid.

[0649] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.19-5.01 (m, 1H), 4.22 (dd, J1 = 3.4, J2 = 11.8Hz, 1H), 4.10-3.99 (m, 3H), 3.63 (br s, 2H), 2.70 (br s, 2H), 2.58 (br s,4H),2.31(dt,J1=2.4,J2=7.4Hz,6H),1.72-1.51(m,14H),1.37-1.18(m,38H),0.97-0.75(m,9H)ppm.

[0650] LCMS:RT=1.810,m / z 712.5[M+H] + .

[0651] Preparation Example 18 Preparation of Compound 21

[0652] Step 1: Synthesis of (2,2-dimethyl-1,3-dioxolane-4-yl)methyl 5-bromopentyl ester

[0653] Reaction formula:

[0654]

[0655] Material ratio:

[0656]

[0657] Operation process:

[0658] (2,2-Dimethyl-1,3-dioxolane-4-yl)methanol (3.32 g, 25.1 mmol) and 5-bromovaleric acid (5.00 g, 27.6 mmol) were dissolved in tetrahydrofuran (35 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.22 g, 37.7 mmol) and 4-dimethylaminopyridine (306 mg, 2.51 mmol) were added in sequence. The mixture was reacted at 25 ° C. under nitrogen protection for 12 hours. The reaction was monitored by TLC. 50 mL of water was added and the mixture was extracted with dichloromethane twice, 50 mL each time. The organic phase was concentrated and purified by column chromatography to obtain a bright yellow liquid (2,2-dimethyl-1,3-dioxolane-4-yl)methyl 5-bromopentyl ester (6.31 g, 85.1% yield).

[0659] Step 2: Synthesis of 2,3-dihydroxypropyl 5-bromopentyl ester

[0660] Reaction formula:

[0661]

[0662] Material ratio:

[0663]

[0664] Operation process:

[0665] Dissolve (2,2-dimethyl-1,3-dioxolane-4-yl)methyl 5-bromopentyl ester (5.80 g, 19.7 mmol) in tetrahydrofuran (58 mL). Add hydrochloric acid (5.80 mL, 17.4 mmol). React at 25°C for 12 hours. TLC monitoring indicates the formation of new spots. Add 10 mL of aqueous sodium bicarbonate solution, extract with ethyl acetate twice (20 mL each time), and concentrate the organic phase. Column chromatography yields the crude product as a yellow liquid, 2,3-dihydroxypropyl 5-bromopentyl ester (3.50 g, 69.8% yield).

[0666] Step 3: Synthesis of 1-[(5-bromovaleryl)oxy]-3-(octanoyloxy)propan-2-yloctyl ester

[0667] Reaction formula:

[0668]

[0669] Material ratio:

[0670]

[0671] Operation process:

[0672] 2,3-Dihydroxypropyl 5-bromopentyl ester (3.50 g, 13.7 mmol) and octanoic acid (4.95 g, 34.3 mmol) were dissolved in dichloromethane (70 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.89 g, 41.16 mmol) and 4-dimethylaminopyridine (335 mg, 2.74 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After addition of 10 mL of water, the mixture was extracted with dichloromethane twice, each time using 10 mL. The organic phase was concentrated. Purification by column chromatography afforded 1-[(5-bromovaleroyl)oxy]-3-(octanoyloxy)propan-2-yloctyl ester (6.14 g, 88.1% yield), a colorless liquid.

[0673] Step 4: Synthesis of heptadecane-9-yl 8-({5-[2,3-di(octanoyloxy)propoxy]-5-oxypentylene}(2-hydroxyethyl)amino)octyl ester

[0674] Reaction formula:

[0675]

[0676] Material ratio:

[0677]

[0678] Operation process:

[0679] 1-[(5-bromovaleroyl)oxy]-3-(octanoyloxy)propan-2-yl octyl ester (1.00 g, 2.26 mmol) was dissolved in acetonitrile (5.0 mL). Heptadec-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (1.26 g, 2.49 mmol), potassium carbonate (1.10 g, 7.92 mmol), potassium iodide (450 mg, 2.72 mmol), and tetrahydrofuran (5 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadec-9-yl 8-({5-[2,3-di(octanoyloxy)propoxy]-5-oxypentylene}(2-hydroxyethyl)amino)octyl ester (230 mg, 10.4% yield) as a bright yellow liquid.

[0680] 1H NMR (400MHz, CHLOROFORM-d) δ=5.34-5.25(m,1H),4.88(quin,J=6.2Hz,1H),4.37-4.28(m,2H),4.17(dd,J1=6.0,J2=12.0Hz,2H),3.59-3.51(m,2H),2.60(br s,2H),2.47(br d,J=1.0Hz,4H),2.38-2.27(m,8H),1.68-1.59(m,10H),1.55-1.42(m,9H),1.37-1.23(m,48H),0.94-0.85(m,12H)ppm.

[0681] LCMS:RT=2.062,m / z 868.6[M+H] + .

[0682] Preparation Example 19 Preparation of Compound 32

[0683] Step 1: Synthesis of 5-(benzyloxy)pentan-1-ol

[0684] Reaction formula:

[0685]

[0686] Material ratio:

[0687] Material Name Molecular weight Feed ratio Feeding amount mmol Pentylene glycol 104 1.0 eq 15.0g 144mmol Bianium Bromide 170 1.0eq 24.6g 144mmol Sodium hydride 24 1.2eq 6.91g 173mmol N,N-dimethylformamide - - 300mL -

[0688] Operation process:

[0689] Pentanediol (15.0 g, 144 mmol) and sodium hydride (6.91 g, 173 mmol, 60%) were dissolved in N,N-dimethylformamide (150 mL). The mixture was stirred at 20°C under nitrogen for 0.5 hours. A solution of benzyl bromide (24.6 g, 144 mmol) in N,N-dimethylformamide (150 mL) was slowly added dropwise at 0°C. The mixture was allowed to react at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was washed with water, extracted, and concentrated. The crude product was purified by column chromatography to yield 5-(benzyloxy)pentan-1-ol (14.8 g, 52.9% yield) as a colorless liquid.

[0690] Step 2: Synthesis of 5-(benzyloxy)valeraldehyde

[0691] Reaction formula:

[0692]

[0693] Material ratio:

[0694] Material Name Molecular weight Feed ratio Feeding amount mmol 5-(Benzyloxy)pentan-1-ol 194 1.0 eq 14.8g 76.2mmol Des Martin 424 1.2eq 38.8g 91.4mmol dichloromethane - - 150mL -

[0695] Operation process:

[0696] 5-(Benzyloxy)pentan-1-ol (14.5 g, 76.2 mmol) was dissolved in dichloromethane (150 mL), and Dess-Martin (38.8 g, 91.4 mmol) was added at 0°C. The reaction was allowed to proceed at 20°C for 2 hours under nitrogen. Completion of the reaction was monitored by TLC. The reaction solution was slowly poured into a saturated aqueous sodium bicarbonate solution at 0°C to quench the reaction. The mixture was washed with water, extracted, and concentrated. Column chromatography of the crude product afforded 5-(benzyloxy)pentanal (10.1 g, 57.5% yield) as a colorless liquid.

[0697] Step 3: Synthesis of 1-(benzyloxy)dodecan-5-ol

[0698] Reaction formula:

[0699]

[0700] Material ratio:

[0701] Material Name Molecular weight Feed ratio Feeding amount mmol 5-(Benzyloxy)pentanal 192 1.0 eq 10.0g 52.0mmol Heptylmagnesium bromide 202 1.05eq 11.1g 54.6mmol Tetrahydrofuran - - 100mL -

[0702] Operation process:

[0703] 5-(Benzyloxy)pentanal (10.0 g, 52.0 mmol) was dissolved in tetrahydrofuran (100 mL). A solution of heptylmagnesium bromide (11.1 g, 54.6 mmol) in tetrahydrofuran was slowly added dropwise at 0°C. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to afford 1-(benzyloxy)dodecan-5-ol (10.0 g, 59.2% yield), a colorless liquid.

[0704] Step 4: Synthesis of 1-(benzyloxy)dodec-5-ylhexyl ester

[0705] Reaction formula:

[0706]

[0707] Material ratio:

[0708]

[0709] Operation process:

[0710] 1-(Benzyloxy)dodecan-5-ol (10.0 g, 34.2 mmol) was dissolved in dichloromethane (100 mL). Hexanoic acid (5.16 g, 44.5 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (9.83 g, 51.3 mmol), and 4-dimethylaminopyridine (418 mg, 3.42 mmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodecan-5-ylhexyl ester (9.00 g, 67.4% yield) as a colorless liquid.

[0711] Step 5: Synthesis of 1-hydroxydodecane-5-ylhexyl ester

[0712] Reaction formula:

[0713]

[0714] Material ratio:

[0715] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodec-5-ylhexyl ester 390 1.0 eq 9.00g 23.0mmol Palladium / Carbon 106 0.1eq 2.50g 2.40mmol hydrogen - (40Psi) - - Methanol - 90mL -

[0716] Operation process:

[0717] 1-(Benzyloxy)dodec-5-ylhexyl ester (9.00 g, 23.0 mmol) was dissolved in methanol (90 mL). Palladium / carbon (2.50 g, 2.40 mmol) was added under a nitrogen atmosphere. The reaction was then allowed to proceed at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was filtered and concentrated. The crude product was purified by column chromatography to afford 1-hydroxydodec-5-ylhexyl ester (7.20 g, 92.9% yield) as a colorless liquid.

[0718] Step 6: Synthesis of 1-[(5-bromovaleryl)oxy]dodec-5-ylhexyl ester

[0719] Reaction formula:

[0720]

[0721] Material ratio:

[0722]

[0723] Operation process:

[0724] 1-Hydroxydodec-5-ylhexyl ester (2.00 g, 6.66 mmol) was dissolved in dichloromethane (20 mL). 5-Bromopentanoic acid (1.57 g, 8.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.91 g, 9.98 mmol), and 4-dimethylaminopyridine (81.3 mg, 666 μmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-[(5-bromovaleroyl)oxy]dodec-5-ylhexyl ester (2.10 g, 68.1% yield) as a colorless liquid.

[0725] Step 7: Synthesis of heptadecane-9-yl 8-[(5-{[5-(hexanoyloxy)dodecyl]oxy}-5-oxypentylidene)(2-hydroxyethyl)amino]octyl ester

[0726] Reaction formula:

[0727]

[0728] Material ratio:

[0729]

[0730] Operation process:

[0731] The reaction was carried out in parallel in batch 1.33. 1-[(5-bromovaleroyl)oxy]dodec-5-yl hexyl ester (1.65 g, 3.57 mmol) was dissolved in acetonitrile (12 mL). Heptadecan-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (1.50 g, 3.40 mmol), potassium carbonate (1.64 g, 11.9 mmol), potassium iodide (676 mg, 4.07 mmol), and tetrahydrofuran (3 mL) were added sequentially. The mixture was reacted at 70°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadecan-9-yl 8-[(5-{[5-(hexanoyloxy)dodecyl]oxy}-5-oxypentylene)(2-hydroxyethyl)amino]octyl ester (800 mg, 21.6% yield) as a pale yellow liquid.

[0732] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.87 (sex, J = 6.0Hz, 2H), 4.05 (t, J = 6.6Hz, 2H), 3.60 (t, J = 5.0Hz, 2H), 2.66 (br t,J=5.0Hz,2H),2.61-2.49(m,4H),2.35-2.25(m,6H),1.67-1.58(m,8H),1.57-1.45(m,12H),1.38-1.29(m,14H),1.26(br s,32H),0.99-0.78(m,12H)ppm.

[0733] LCMS:RT=2.135,m / z 824.6[M+H] + .

[0734] Preparation Example 20 Preparation of Compound 48

[0735] Step 1: Synthesis of 4-(benzyloxy)butan-1-ol

[0736] Reaction formula:

[0737]

[0738] Material ratio:

[0739]

[0740]

[0741] Operation process:

[0742] Butane-1,4-diol (5.00 g, 55.5 mmol) was dissolved in N,N-dimethylformamide (100 mL), and sodium hydroxide (3.33 g, 83.2 mmol) was added at 25°C. The mixture was reacted at 25°C for half an hour. Benzyl bromide (9.49 g, 55.5 mmol) was added to the reaction solution, and the mixture was reacted at 25°C for half an hour under nitrogen protection. The reaction was completed by TLC monitoring. 1000 mL of water was added, and the mixture was extracted twice with 500 mL of ethyl acetate each time. The organic phase was concentrated and purified by column chromatography to obtain a colorless liquid 4-(benzyloxy)butane-1-ol (9.60 g, 96.0% yield).

[0743] Step 2: Synthesis of 4-(benzyloxy)butyraldehyde

[0744] Reaction formula:

[0745]

[0746] Material ratio:

[0747] Material Name Molecular weight Feed ratio Feeding amount mmol 4-(Benzyloxy)butan-1-ol 180 1.0 eq 9.60g 53.3mmol Dess-Martin periodinane 423 1.2eq 27.1mg 63.9mmol dichloromethane - - 150mL -

[0748] Operation process:

[0749] Dissolve 4-(Benzyloxy)butan-1-ol (9.60 g, 53.3 mmol) in dichloromethane (150 mL). Add Dess-Martin periodinane (27.1 g, 63.9 mmol) at 0°C. Allow to react at 20°C for 2 hours. TLC monitoring indicates the formation of new spots. Add 100 mL of aqueous sodium bicarbonate solution, extract twice with 200 mL of dichloromethane, and concentrate the organic phase. Column chromatography yields the crude product as a colorless liquid, 4-(benzyloxy)butyraldehyde (6.32 g, 66.5% yield).

[0750] Step 3: Synthesis of 1-(benzyloxy)dodecan-4-ol

[0751] Reaction formula:

[0752]

[0753] Material ratio:

[0754] Material Name Molecular weight Feed ratio Feeding amount mmol 4-(Benzyloxy)butyraldehyde 178 1.0 eq 6.32g 35.5mmol Octylmagnesium bromide 216 1.2eq 9.25g 42.6mmol Tetrahydrofuran - - 64mL -

[0755] Operation process:

[0756] 4-(Benzyloxy)butyraldehyde (6.32 g, 35.5 mmol) was dissolved in tetrahydrofuran (64 mL). (Octyl)magnesium bromide (9.25 g, 42.6 mmol) was added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. 10 mL of saturated aqueous ammonium chloride was added, and the mixture was extracted with ethyl acetate twice, each time using 20 mL. The organic phase was concentrated. Purification by column chromatography afforded 1-(benzyloxy)dodecan-4-ol (7.14 g, 68.1% yield), a colorless liquid.

[0757] Step 4: Synthesis of 1-(benzyloxy)dodec-4-ylheptyl ester

[0758] Reaction formula:

[0759]

[0760] Material ratio:

[0761]

[0762] Operation process:

[0763] 1-(Benzyloxy)dodecan-4-ol (7.14 g, 24.4 mmol) and heptanoic acid (4.77 g, 36.6 mmol) were dissolved in dichloromethane (140 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.02 g, 36.6 mmol) and 4-dimethylaminopyridine (298 mg, 2.44 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. 100 mL of water was added, and the mixture was extracted with dichloromethane twice, each time using 100 mL. The organic phase was concentrated. Purification by column chromatography afforded 1-(Benzyloxy)dodecan-4-ylheptyl ester (7.13 g, 72.1% yield) as a colorless liquid.

[0764] Step 5: Synthesis of 1-hydroxydodec-4-ylheptyl ester

[0765] Reaction formula:

[0766]

[0767] Material ratio:

[0768] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodec-4-ylheptyl ester 404 1.0 eq 7.13g 17.6mmol Palladium on carbon 105 0.2eq 3.75g 3.52mmol Methanol - - 74mL -

[0769] Operation process:

[0770] Dissolve 1-(Benzyloxy)dodec-4-ylheptyl ester (7.13 g, 17.6 mmol) in methanol (74 mL), add palladium on carbon (3.75 g, 3.52 mmol), and react under hydrogen at 35°C, 40 psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Column chromatography yields the crude product as a colorless liquid, 1-hydroxydodec-4-ylheptyl ester (5.07 g, 91.4% yield).

[0771] Step 6: Synthesis of 1-[(5-bromovaleryl)oxy]dodec-4-ylheptyl ester

[0772] Reaction formula:

[0773]

[0774] Material ratio:

[0775]

[0776] Operation process:

[0777] 1-Hydroxydodec-4-ylheptyl ester (5.07 g, 16.1 mmol) and 5-bromovaleric acid (3.21 g, 17.7 mmol) were dissolved in dichloromethane (100 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.64 g, 24.2 mmol) and 4-dimethylaminopyridine (196 mg, 1.61 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. 100 mL of water was added, and the mixture was extracted with dichloromethane twice, each time using 100 mL. The organic phase was concentrated. Purification by column chromatography gave 1-[(5-bromovaleryl)oxy]dodec-4-ylheptyl ester (4.20 g, 54.5% yield) as a bright yellow liquid.

[0778] Step 7: Synthesis of heptadecane-9-yl 8-[(5-{[4-(heptanoyloxy)dodecyl]oxy}-5-oxypentylidene)(2-hydroxyethyl)amino]octyl ester

[0779] Reaction formula:

[0780]

[0781] Material ratio:

[0782]

[0783] Operation process:

[0784] 1-[(5-bromovaleroyl)oxy]dodec-4-yl heptyl ester (594 mg, 1.25 mmol) was dissolved in acetonitrile (3.0 mL), and heptadecan-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (500 mg, 1.13 mmol), potassium carbonate (547 mg, 3.96 mmol), potassium iodide (225 mg, 1.36 mmol), and tetrahydrofuran (2 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadecan-9-yl 8-[(5-{[4-(heptanoyloxy)dodecyl]oxy}-5-oxypentylene)(2-hydroxyethyl)amino]octyl ester (670 mg, 66.4% yield) as a bright yellow liquid.

[0785] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.89 (td, J = 6.2, 12.8Hz, 2H), 4.15-4.00 (m, 2H), 3.57 (t, J = 5.2Hz, 2H), 2.63 (br t,J=5.2Hz,2H),2.51(td,J1=7.4,J2=12.0Hz,4H),2.35-2.26(m,6H),1.69-1 .57(m,11H),1.56-1.45(m,10H),1.35-1.22(m,48H),0.97-0.83(m,12H)ppm.

[0786] LCMS:RT=2.183,m / z 839.3[M+H] + .

[0787] Preparation Example 21 Preparation of Compound 55

[0788] Step 1: Synthesis of 1-[(6-bromohexanoyl)oxy]dodec-2-yl (4E)-oct-4-enyl ester

[0789] Reaction formula:

[0790]

[0791] Material ratio:

[0792]

[0793] Operation process:

[0794] To the reaction flask were added dichloromethane (30.0 mL), 1-hydroxydodec-2-yl (4E)-oct-4-enol (3.00 g, 9.19 mmol), 6-bromohexanoic acid (2.15 g, 11.0 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.64 g, 13.7 mmol), and 4-dimethylaminopyridine (112 mg, 918 μmol). The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction solution was concentrated and purified by column chromatography to afford 1-[(6-bromohexanoyl)oxy]dodec-2-yl (4E)-oct-4-enol (3.27 g, 70.6% yield) as a colorless liquid.

[0795] Step 2: Synthesis of 1-({6-[(2-hydroxyethyl)[6-({2-[(4E)-oct-4-enoxy]dodecyl}oxy)-6-oxyylidenehexyl]amino]hexanoyl}oxy)dodec-2-yl(4E)-oct-4-enyl ester

[0796] Reaction formula:

[0797]

[0798] Material ratio:

[0799]

[0800] Operation process:

[0801] 1-[(6-bromohexanoyl)oxy]dodec-2-yl (4E)-oct-4-enate (1.00 g, 1.99 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (58.2 mg, 953 μmol), potassium carbonate (960 mg, 6.95 mmol), potassium iodide (395 mg, 2.38 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({6-[(2-hydroxyethyl)[6-({2-[(4E)-oct-4-enoxy]dodecyl}oxy)-6-oxyylidenehexyl]amino]hexanoyl}oxy)dodec-2-yl (4E)-oct-4-enate (669 mg, 31.7% yield) as a colorless liquid.

[0802] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.62-5.29 (m, 4H), 5.17-4.85 (m, 2H), 4.23 (dd, J1 = 3.4, J2 = 11.8Hz, 2H), 4.03 (dd, J1 = 6.4, J2 = 11.8Hz, 2H), 3.58 (br s,2H),2.64(br s,2H),2.52(br t,J=6.4Hz,4H),2.40-2.27(m,12H),1.96(q,J=6.8Hz,4H),1.67-1.47(m,12H),1.37-1.18(m,40H),0.99-0.75(m,12H)ppm.

[0803] LCMS:RT=2.070,m / z 906.5[M+H] + .

[0804] Preparation Example 22 Preparation of Compound 57

[0805] Step 1: Synthesis of ethyl 4-oct-4-enyl ester

[0806] Reaction formula:

[0807]

[0808] Material ratio:

[0809] Material Name Molecular weight Feed ratio Feeding amount mmol 1-Hexen-3-ol 100 1.00 eq 15.0g 150mmol Triethyl orthoacetate 162 3.00eq 72.9g 449mmol Propionic acid 74 0.1eq 1.11g 15.0mmol

[0810] Operation process:

[0811] 1-Hexen-3-ol (15.0 g, 150 mmol) was dissolved in triethyl orthoacetate (72.9 g, 449 mmol), and propionic acid (1.11 g, 15.0 mmol) was added. The mixture was reacted at 130°C under nitrogen for 6 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield ethyl 4-oct-4-enyl ester (24.1 g, 70.9% yield), a colorless liquid.

[0812] Step 2: Synthesis of oct-4-enoic acid

[0813] Reaction formula:

[0814]

[0815] Material ratio:

[0816] Material Name Molecular weight Feed ratio Feeding amount mmol Ethyl 4-oct-4-enol 170 1.00 eq 22.0g 129mmol Lithium hydroxide monohydrate 42 3.00eq 16.3g 388mmol Methanol - - 220mL -

[0817] Operation process:

[0818] Dissolve ethyl 4-oct-4-enyl ester (22.0 g, 129 mmol) in methanol (220 mL), then add lithium hydroxide monohydrate (16.3 g, 388 mmol). Finally, react at 20°C under nitrogen for 2 hours. Completion of the reaction was monitored by TLC. The reaction solution was acidified, washed with water, extracted, and concentrated. Column chromatography of the crude product afforded oct-4-enoic acid (16.5 g, 81.6% yield) as a colorless liquid.

[0819] Step 3: Synthesis of 1-hydroxydodec-2-yloct-4-enol

[0820] Reaction formula:

[0821]

[0822] Material ratio:

[0823] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1.0 eq 20.0g 108mmol Oct-4-enoic acid 142 1.05eq 16.2g 113mmol Pyridine 79 0.07 eq 600mg 7.60mmol Ferric chloride 160 0.025 eq 440mg 2.71mmol

[0824] Operation process:

[0825] 1,2-Epoxydodecane (20.0 g, 108 mmol), oct-4-enoic acid (16.2 mg, 113 mmol), pyridine (600 mg, 7.60 mmol), and ferric chloride (440 mg, 2.71 mmol) were added sequentially to a reaction flask and reacted at 30°C for 12 hours. The reaction was monitored by TLC, and the mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography to obtain 1-hydroxydodec-2-yloct-4-enyl ester (18.9 g, 53.3% yield) as a white solid.

[0826] Step 4: Synthesis of 1-[(8-bromooctanoyl)oxy]dodec-2-yloct-4-enate

[0827] Reaction formula:

[0828]

[0829] Material ratio:

[0830]

[0831]

[0832] Operation process:

[0833] 1-Hydroxydodec-2-yloct-4-enate (2.00 g, 6.13 mmol) was dissolved in dichloromethane (20 mL). 8-Bromooctanoic acid (1.78 g, 7.96 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.76 g, 9.19 mmol), and 4-dimethylaminopyridine (74.8 mg, 613 umol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain 1-[(8-bromooctanoyl)oxy]dodec-2-yloct-4-enate (2.20 g, 67.6% yield), a colorless liquid.

[0834] Step 5: Synthesis of 1-({8-[(2-hydroxyethyl)(8-{[2-(oct-4-enoxy)dodecyl]oxy}-8-oxyoctylene)amino]octanoyl}oxy)dodec-2-yloct-4-enyl ester

[0835] Reaction formula:

[0836]

[0837] Material ratio:

[0838]

[0839] Operation process:

[0840] 1-[(8-Bromooctanoyl)oxy]dodec-2-yloct-4-enate (1.50 g, 2.28 mmol) was dissolved in acetonitrile (10 mL). Ethanolamine (84.5 mg, 1.38 mmol), potassium carbonate (1.36 g, 9.88 mmol), potassium iodide (562 mg, 3.39 mmol), and tetrahydrofuran (5 mL) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. The crude product was purified by column chromatography to yield 1-({8-[(2-hydroxyethyl)(8-{[2-(oct-4-enoxy)dodecyl]oxy}-8-oxyylideneoctyl)amino]octanoyl}oxy)dodec-2-yloct-4-enate (800 mg, 26.8% yield) as a pale yellow liquid.

[0841] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.58-5.32 (m, 4H), 5.11-5.04 (m, 2H), 4.22 (dd, J1 = 3. 4,J2=11.8Hz,2H),4.03(dd,J1=6.6,J2=11.8Hz,2H),3.56(t,J=5.2Hz,2H),2.61(br t,J=5.0Hz,2H),2.48(br t,J=7.4Hz,4H),2.40-2.35(m,4H),2.30(br t,J=7.4Hz,8H),1.95(q,J=6.6Hz,4H),1.65-1.54(m,8H),1.50-1.43(m,4H),1.41-1.25(m,48H),0.91-0.85(m,12H)ppm.

[0842] LCMS:RT=2.132,m / z 962.7[M+H] + .

[0843] Preparation Example 23 Preparation of Compound 11

[0844] Step 1: Preparation of 11-1

[0845] Reaction formula:

[0846]

[0847] Material ratio:

[0848] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1eq 5g 27 8-Bromooctanoic acid 223 1.1 eq 6.7g 30 Ferric chloride 162 0.025 eq 220mg 1.35 Pyridine 79 0.0125eq 53mg 0.68

[0849] Operation process:

[0850] 8-Bromooctanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to the reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 4:1) indicated completion of the reaction (product rf value 0.5). Purification by column chromatography yielded 7.5 g of a colorless oil.

[0851] Step 2: Preparation of 11-2

[0852] Reaction formula:

[0853]

[0854] Material ratio:

[0855] Material Name Molecular weight Feed ratio Feeding amount mmol 11-1 407.43 1eq 7.1g 17.5 bitter 144 1.1 eq 2.8g 19.2 EDCI 192 1.5eq 5g 26 DMAP 122 0.15 eq 320mg 2.6 dichloromethane - - 80mL -

[0856] Operation process:

[0857] To the reaction flask, 11-1, octanoic acid, EDCI, DMAP, and dichloromethane were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 6 g of a colorless oil.

[0858] Step 3: Preparation of compound 11

[0859] Reaction formula:

[0860]

[0861] Material ratio:

[0862] Material Name Molecular weight Feed ratio Feeding amount mmol 11-2 533.63 2.2 eq 4.56g 8.55 Ethanolamine 61 1 eq 237mg 3.9 K2CO3 138 3 eq 1.6g 11.6 KI 166 2 eq 1.3g 7.8 Acetonitrile - - 40mL -

[0863] Operation process:

[0864] To the reaction flask, 11-2, ethanolamine, K2CO3, KI, and acetonitrile were added and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was concentrated, then diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.8 g of a colorless oil.

[0865] 1H NMR(600MHz,Chloroform-d)δ5.07(qd,J=6.9,3.3Hz,1H),4.20(dd,J=11.8,3.3Hz,1H),4.01(dd,J=11.8,6.8Hz,1H),3.57(t,J=5.3Hz,1H),2.63(t, J=5.3Hz,1H),2.51(t,J=7.6Hz,2H),2.29(td,J=7.5,1.9Hz,4H),1.63–1.5 1(m,6H),1.47(p,J=7.4Hz,2H),1.32–1.22(m,30H),0.87(t,J=6.8Hz,6H).

[0866] MS (ES+) m / z): 967.63 (M+H) + .

[0867] Preparation Example 24 Preparation of Compound 19

[0868] Step 1: Synthesis of 1-(benzyloxy)dodecan-3-ol

[0869] Reaction formula:

[0870]

[0871] Material ratio:

[0872] Material Name Molecular weight Feed ratio Feeding amount mmol 3-(Benzyloxy)propanal 164 1.00 eq 6.00g 36.5mmol Nonylmagnesium bromide 230 1.20eq 10.1g 43.8mmol Tetrahydrofuran - - 60.0mL -

[0873] Operation process:

[0874] Dissolve 3-(Benzyloxy)propanal (6.00 g, 36.5 mmol) in tetrahydrofuran (60.0 mL). Slowly add a solution of nonylmagnesium bromide (10.1 g, 43.8 mmol) in tetrahydrofuran at 0°C. Incubate at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. Column chromatography of the crude product affords 1-(benzyloxy)dodecan-3-ol (5.87 g) as a colorless liquid.

[0875] Step 2: Synthesis of 1-(benzyloxy)dodec-3-yloctyl ester

[0876] Reaction formula:

[0877]

[0878] Material ratio:

[0879]

[0880] Operation process:

[0881] Dissolve 1-(Benzyloxy)dodecan-3-ol (5.87 g, 20.0 mmol) and n-octanoic acid (3.47 g, 24.0 mmol) in dichloromethane (58.0 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.77 g, 30.1 mmol) and 4-dimethylaminopyridine (245 mg, 2.01 mmol) sequentially. The mixture is reacted at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC, and the reaction solution is concentrated. Column chromatography of the crude product affords 1-(Benzyloxy)dodecan-3-yloctyl ester (6.00 g) as a colorless liquid.

[0882] Step 3: Synthesis of 1-hydroxydodecane-3-yl octyl ester

[0883] Reaction formula:

[0884]

[0885] Material ratio:

[0886] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodec-3-yloctyl ester 418 1.0 eq 6.00g 14.3mmol Palladium / Carbon 106 0.20eq 3.05g 2.87mmol hydrogen - (40Psi) - - Methanol - - 60.0mL -

[0887] Operation process:

[0888] Dissolve 1-(Benzyloxy)dodec-3-yloctyl ester (6.00 g, 14.3 mmol) in methanol (60.0 mL). Add palladium / carbon (3.05 g, 2.87 mmol) under argon. Incubate at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Monitor the reaction by TLC. Filter and concentrate the reaction mixture to obtain a colorless liquid, 1-hydroxydodec-3-yloctyl ester (4.60 g).

[0889] Step 4: Synthesis of 1-[(6-bromohexanoyl)oxy]dodec-3-yloctyl ester

[0890] Reaction formula:

[0891]

[0892] Material ratio:

[0893]

[0894]

[0895] Operation process:

[0896] Dissolve 1-hydroxydodec-3-yloctyl ester (2.00 g, 6.09 mmol) in dichloromethane (15.0 mL). Add 6-bromohexanoic acid (1.42 g, 7.31 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.75 g, 9.13 mmol), and 4-dimethylaminopyridine (74.3 mg, 608 μmol) in this order. The mixture is reacted at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. The crude product is purified by column chromatography to yield 1-[(6-bromohexanoyl)oxy]dodec-3-yloctyl ester (2.78 g), a colorless liquid.

[0897] Step 5: Synthesis of 1-({6-[(2-hydroxyethyl)(6-{[3-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodec-3-yloctyl ester

[0898] Reaction formula:

[0899]

[0900] Material ratio:

[0901]

[0902] Operation process:

[0903] 1-[(6-bromohexanoyl)oxy]dodec-3-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (57.9 mg, 949 μmol), potassium carbonate (478 mg, 3.46 mmol), potassium iodide (197 mg, 1.19 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({6-[(2-hydroxyethyl)(6-{[3-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodec-3-yloctyl ester (603 mg) as a colorless liquid.

[0904] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.08-4.88(m,2H),4.17-3.98(m,4H),3.58(brs,2H),2.64(br s,2H),2.52(br s,4H),2.30(q,J=7.4Hz,8H),1.94-1.80(m,4H),1.69-1.49(m,16H),1.34-1.18(m,48H),0.89(t,J=6.8Hz,12H)ppm.

[0905] LCMS:RT=2.169,m / z 910.6[M+H] + .

[0906] Preparation Example 25 Preparation of Compound 20

[0907] Step 1: Synthesis of 1-[(5-bromovaleryl)oxy]dodec-3-yloctyl ester

[0908] Reaction formula:

[0909]

[0910] Material ratio:

[0911]

[0912] Operation process:

[0913] Dissolve 1-hydroxydodec-3-yloctyl ester (1.00 g, 3.04 mmol) and 5-bromovaleric acid (661 mg, 3.65 mmol) in dichloromethane (10.0 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (875 mg, 4.57 mmol) and 4-dimethylaminopyridine (37.1 mg, 304 μmol) sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC, and the reaction solution was concentrated. Column chromatography of the crude product afforded 1-[(5-bromovaleryl)oxy]dodec-3-yloctyl ester (1.40 g) as a colorless liquid.

[0914] Step 2: Synthesis of heptadec-9-yl 8-[(2-hydroxyethyl)(5-{[3-(octanoyloxy)dodecyl]oxy}-5-oxypentylene)amino]octyl ester

[0915] Reaction formula:

[0916]

[0917] Material ratio:

[0918]

[0919] Operation process:

[0920] Dissolve 1-[(5-bromovaleroyl)oxy]dodec-3-yl octyl ester (1.17 g, 2.38 mmol) in acetonitrile (10.0 mL). Add heptadec-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (1.00 g, 2.26 mmol), potassium carbonate (1.10 g, 7.92 mmol), potassium iodide (450 mg, 2.72 mmol), and tetrahydrofuran (5.00 mL) in this order. The mixture is reacted at 75°C under nitrogen for 12 hours. The reaction is monitored by TLC and concentrated. Purification by column chromatography affords a colorless liquid, heptadec-9-yl 8-[(2-hydroxyethyl)(5-{[3-(octanoyloxy)dodecyl]oxy}-5-oxypentylene)amino]octyl ester (1.50 g).

[0921] 1 H NMR (400MHz, CHLOROFORM-d) δ=5.06-4.92(m,1H),4.87(quin,J=6.2Hz,1H),4.18-4.01(m,2H),3.56(br t,J=5.0Hz,2H),2.62(br s,2H),2.56-2.43(m,4H),2.37-2.24(m,6H),1.94-1.80(m,2H),1.64-1.44(m,16H),1.33-1.15(m,52H),0.88(t,J=6.8Hz,12H)ppm.

[0922] LCMS:RT=2.325,m / z 874.6[M+23] + .

[0923] Preparation Example 26 Preparation of Compound 22

[0924] Step 1: Preparation of 22-1

[0925] Reaction formula:

[0926]

[0927] Material ratio:

[0928] Material Name Molecular weight Feed ratio Feeding amount mmol 1,2-Epoxydodecane 184 1eq 5g 27 6-Bromohexanoic acid 195 1.1eq 5.85g 30 Ferric chloride 162 0.025 eq 220mg 1.35 Pyridine 79 0.0125eq 53mg 0.68

[0929] Operation process:

[0930] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to the reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 4:1) indicated completion of the reaction (product rf value 0.5). Purification by column chromatography yielded 7 g of a colorless oil.

[0931] Step 2: Preparation of 22-2

[0932] Reaction formula:

[0933]

[0934] Material ratio:

[0935] Material Name Molecular weight Feed ratio Feeding amount mmol 22-1 379.38 1eq 6.6g 17.5 Hexanoic acid 116 1.1 eq 2.2g 19.2 EDCI 192 1.5eq 5g 26 DMAP 122 0.15 eq 320mg 2.6 dichloromethane - - 80mL -

[0936] Operation process:

[0937] To the reaction flask, 22-1, hexanoic acid, EDCI, DMAP, and dichloromethane were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) indicated completion of the reaction (product rf value 0.6). The reaction mixture was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 5.6 g of a colorless oil.

[0938] Step 3: Preparation of compound 22

[0939] Reaction formula:

[0940]

[0941] Material ratio:

[0942] Material Name Molecular weight Feed ratio Feeding amount mmol 22-2 477.52 1.1eq 4.1g 8.55 LQ001-2 441.74 1 eq 3.4g 7.8 K2CO3 138 1.5eq 1.6g 11.6 KI 166 1 eq 1.3g 7.8 Acetonitrile - - 40mL -

[0943] Operation process:

[0944] 22-2, LQ001-2, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) indicated completion of the reaction (product rf value 0.5). The reaction mixture was concentrated, then diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.5 g of a colorless oil.

[0945] 1H NMR (600MHz, Chloroform-d) δ5.08 (qd, J=6.9, 3.2Hz, 1H), 4.85 (p, J=6.2Hz, 1H) ,4.22(dd,J=11.8,3.3Hz,1H),4.01(dd,J=11.8,6.8Hz,1H),3.57(t,J=5.3Hz,2H ),2.62(t,J=5.3Hz,2H),2.50(q,J=7.5Hz,4H),2.32–2.24(m,6H),1.62(p,J=7. 4Hz, 6H), 1.57–1.43 (m, 10H), 1.33–1.22 (m, 52H), 0.88 (dt, J=13.6, 6.8Hz, 12H).

[0946] MS (ES+) m / z): 839.38 (M+H) + .

[0947] Preparation Example 27 Preparation of Compound 30

[0948] Step 1: Synthesis of 5-(hexanoyloxy)dodecyl 6-bromohexyl ester

[0949] Reaction formula:

[0950]

[0951] Material ratio:

[0952]

[0953] Operation process:

[0954] 1-Hydroxydodec-5-ylhexyl ester (2.00 g, 6.66 mmol) and 6-bromohexanoic acid (1.69 g, 8.65 mmol) were dissolved in dichloromethane (20 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.91 g, 9.98 mmol) and 4-dimethylaminopyridine (8.3 mg, 0.665 mmol) were added in sequence. Finally, the mixture was reacted at 45 ° C under nitrogen protection for 12 hours. The reaction was monitored by TLC. The organic phase was concentrated, washed twice with ethyl acetate, and the organic phase was concentrated. The mixture was purified by column chromatography to obtain a colorless liquid 5-(hexanoyloxy)dodecyl 6-bromohexyl ester (2.30 g).

[0955] Step 2: Synthesis of 5-(hexanoyloxy)dodecyl 6-[(6-{[5-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester

[0956] Reaction formula:

[0957]

[0958] Material ratio:

[0959]

[0960] Operation process:

[0961] 5-(Hexanoyloxy)dodecyl 6-bromohexyl ester (1.00 g, 2.09 mmol) was dissolved in acetonitrile (6.0 mL), and 2-aminoethane-1-ol (57.5 mg, 0.942 mmol), potassium carbonate (868 mg, 6.28 mmol), potassium iodide (521 mg, 3.14 mmol), and tetrahydrofuran (4 mL) were added in sequence. The mixture was reacted at 75° C. under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 5-(hexanoyloxy)dodecyl 6-[(6-{[5-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester (550 mg) as a bright yellow liquid.

[0962] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.90 (quin, J = 6.0Hz, 2H), 4.07 (t, J = 6.4Hz, 4H), 3.67 (br s, 2H), 2.73 (br d, J = 1.2Hz, 2H), 2.62 (br d,J=5.6Hz,3H),2.31(q,J=7.6Hz,8H),1.73-1.48(m,28H),1.42-1.23(m,36H),0.97-0.86(m,12H)ppm.

[0963] LCMS:RT=1.960,m / z 854.6[M+H] + .

[0964] Preparation Example 28 Preparation of Compound 31

[0965] Step 1: Synthesis of 6-(benzyloxy)hexane-1-ol

[0966] Reaction formula:

[0967]

[0968] Material ratio:

[0969] Material Name Molecular weight Feed ratio Feeding amount mmol Hexane-1,6-diol 118 1.0 eq 5.00g 42.3mmol (Bromomethyl)benzene 169 5.0eq 7.24g 42.3mmol Sodium cyanide 24 1.5eq 2.54g 63.4mmol N,N-dimethylformamide - - 100mL -

[0970] Operation process:

[0971] In a reaction flask, hexane-1,6-diol (5.00 g, 42.3 mmol) was dissolved in N,N-dimethylformamide (100 mL). The temperature was lowered to 0°C, and sodium hydride (2.54 g, 63.4 mmol) was added in portions. The mixture was allowed to react at 20°C for 30 minutes. Then, (bromomethyl)benzene (7.24 g, 42.3 mmol) was added, and the mixture was allowed to react at 20°C for 30 minutes. The reaction was monitored by TLC. Water (200 mL) was added to quench the reaction, and the mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography to obtain a colorless liquid 6-(benzyloxy)hexane-1-ol (4.00 g).

[0972] Step 2: Synthesis of 6-(benzyloxy)hexanal

[0973] Reaction formula:

[0974]

[0975] Material ratio:

[0976] Material Name Molecular weight Feed ratio Feeding amount mmol 6-(Benzyloxy)hexan-1-ol 208 1.0 eq 4.00g 19.2mmol Dess-Martin periodinane 423 1.2eq 9.77g 23.0mmol dichloromethane - - 40mL -

[0977] Operation process:

[0978] In a reaction flask, 6-(benzyloxy)hexane-1-ol (4.00 g, 19.2 mmol) was dissolved in dichloromethane (40 mL), cooled to 0°C, and Dess-Martin periodinane (9.77 g, 23.0 mmol) was added in batches. The reaction was carried out at 20°C for 1 hour. The reaction was completed by TLC monitoring. Saturated sodium carbonate solution (100 mL) was added to the reaction to quench the reaction. The reaction was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to give a crude product. The crude product was purified by column chromatography to give a colorless liquid 6-(benzyloxy)hexanal (3.00 g).

[0979] Step 3: Synthesis of 1-(benzyloxy)tridec-6-ol

[0980] Reaction formula:

[0981]

[0982] Material ratio:

[0983] Material Name Molecular weight Feed ratio Feeding amount mmol 6-(Benzyloxy)hexanal 206 1.0 eq 3.00g 14.5mmol n-Heptylmagnesium bromide 203 1.2eq 3.55g 17.4mmol Tetrahydrofuran - - 30mL -

[0984] Operation process:

[0985] Dissolve 6-(Benzyloxy)hexanal (3.00 g, 14.5 mmol) in tetrahydrofuran (30 mL). Slowly add a solution of n-heptylmagnesium bromide (3.55 g, 17.4 mmol) in tetrahydrofuran dropwise. Incubate at 20°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. The crude product is purified by column chromatography to yield 1-(benzyloxy)tridec-6-ol (2.90 g), a colorless liquid.

[0986] Step 4: Synthesis of 1-(benzyloxy)tridec-6-ylhexyl ester

[0987] Reaction formula:

[0988]

[0989] Material ratio:

[0990]

[0991] Operation process:

[0992] In a reaction flask, 1-(Benzyloxy)tridecan-6-ol (2.90 g, 9.46 mmol) was dissolved in tetrahydrofuran (30 mL). Hexanoic acid (1.65 g, 14.1 mmol), 4-dimethylaminopyridine (231 mg, 1.89 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.72 g, 14.1 mmol) were added sequentially. The mixture was reacted at 40°C for 16 hours. After completion of the reaction, the mixture was monitored by TLC. The mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography to obtain 1-(Benzyloxy)tridecan-6-ylhexyl ester (3.10 g), a colorless liquid.

[0993] Step 5: Synthesis of 1-hydroxytridecane-6-ylhexyl ester

[0994] Reaction formula:

[0995]

[0996] Material ratio:

[0997]

[0998] Operation process:

[0999] Dissolve 1-hydroxytridec-6-ylhexyl ester (3.10 g, 7.66 mmol) in methanol (30 mL) and tetrahydrofuran (10 mL). Add palladium / carbon (500 mg, 469 μmol) under a nitrogen atmosphere. Finally, react at 30°C under a hydrogen atmosphere (40 psi) for 12 hours. Monitor the reaction by TLC. Filter the reaction mixture and concentrate. Column chromatography yields the crude product as a colorless liquid, 1-hydroxytridec-6-ylhexyl ester (2.10 g).

[1000] Step 6: Synthesis of 6-(hexanoyloxy)tridecyl 6-bromohexyl ester

[1001] Reaction formula:

[1002]

[1003] Material ratio:

[1004]

[1005] Operation process:

[1006] In a reaction flask, 1-hydroxytridec-6-ylhexyl ester (2.10 g, 6.68 mmol) was dissolved in tetrahydrofuran (22 mL). Hexanoic acid (1.95 g, 10.0 mmol), 4-dimethylaminopyridine (163 mg, 1.34 mmol), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.92 g, 10.0 mmol) were added sequentially. The mixture was allowed to react at 40°C for 16 hours. The reaction was monitored by TLC. The mixture was extracted three times with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product. The crude product was purified by column chromatography to obtain 1-(benzyloxy)tridec-6-ylhexyl ester (3.00 g) as a colorless liquid.

[1007] Step 7: Synthesis of 6-(hexanoyloxy)tridecyl 6-[(6-{[6-(hexanoyloxy)tridecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester

[1008] Reaction formula:

[1009]

[1010] Material ratio:

[1011]

[1012] Operation process:

[1013] 1-(Benzyloxy)tridec-6-ylhexyl ester (2.50 g, 2.09 mmol) was dissolved in acetonitrile (15 mL), and ethanolamine (124 mg, 2.03 mmol), potassium carbonate (2.11 g, 15.2 mmol), potassium iodide (1.01 g, 6.10 mmol), and tetrahydrofuran (15 mL) were added in sequence. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 6-(hexanoyloxy)tridecyl 6-[(6-{[6-(hexanoyloxy)tridecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester (1.30 g) as a yellow liquid.

[1014] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.87 (t, J = 6.2Hz, 2H), 4.05 (t, J = 6.8Hz, 4H), 3.72 (br s,2H),2.87-2.62(m,6H),2.30(td,J1=7.6,J2=11.4Hz,8H),1.69-1.59 (m,16H),1.57-1.48(m,8H),1.40-1.24(m,40H),0.96-0.83(m,12H)ppm.

[1015] LCMS:RT=2.019,m / z 882.6[M+H] + .

[1016] Preparation Example 29 Preparation of Compound 52

[1017] Step 1: Synthesis of 7-(benzyloxy)heptan-1-ol

[1018] Reaction formula:

[1019]

[1020] Material ratio:

[1021] Material Name Molecular weight Feed ratio Feeding amount mmol Heptane-1,7-diol 132 1.00 eq 5.00g 37.8mmol Benzyl bromide 169 1.00eq 6.47g 37.8mmol Sodium Hydrogen 24 1.20eq 1.82g 45.3mmol N,N-dimethylformamide - - 100mL -

[1022] Operation process:

[1023] Dissolve heptane-1,7-diol (5.00 g, 37.8 mmol) in N,N-dimethylformamide (100 mL), add sodium hydrogen hydride (1.82 g, 45.3 mmol) at 25°C, react at 0°C for half an hour, add benzyl bromide (6.47 g, 37.8 mmol) to the reaction solution at 0°C, react at 25°C under nitrogen protection for two hours, monitor the completion of the reaction by TLC, quench, extract twice with ethyl acetate, concentrate the organic phase, and purify by column chromatography to obtain colorless liquid 7-(benzyloxy)heptan-1-ol (4.39 g).

[1024] Step 2: Synthesis of 7-(benzyloxy)heptanal

[1025] Reaction formula:

[1026]

[1027] Material ratio:

[1028] Material Name Molecular weight Feed ratio Feeding amount mmol 7-(Benzyloxy)heptan-1-ol 222 1.00 eq 4.39g 19.7mmol Dess-Martin periodinane 423 1.20eq 16.7g 39.4mmol dichloromethane - - 44.0mL -

[1029] Operation process:

[1030] Dissolve 7-(Benzyloxy)heptan-1-ol (4.39 g, 19.7 mmol) in dichloromethane (44.0 mL). Add Dess-Martin periodinane (16.7 g, 39.4 mmol) at 0°C. Allow to react at 25°C for 2 hours. TLC monitoring indicates the formation of new spots. Add 200 mL of aqueous sodium bicarbonate solution, extract with dichloromethane, and concentrate the organic phase. Column chromatography yields the crude product as a colorless liquid, 7-(benzyloxy)heptanal (2.50 g).

[1031] Step 3: Synthesis of 12-(benzyloxy)dodecan-6-ol

[1032] Reaction formula:

[1033]

[1034] Material ratio:

[1035] Material Name Molecular weight Feed ratio Feeding amount mmol 7-(Benzyloxy)heptanal 220 1.00 eq 2.50g 11.3mmol Bromo(pentyl)magnesium 173 1.20eq 2.39g 13.6mmol Tetrahydrofuran - - 25.0mL -

[1036] Operation process:

[1037] 7-(Benzyloxy)heptanal (2.50 g, 11.3 mmol) was dissolved in tetrahydrofuran (25.0 mL). (Amyl)magnesium bromide (2.39 g, 13.6 mmol) was added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. 100 mL of saturated aqueous ammonium chloride was added, and the mixture was extracted with ethyl acetate. The organic phase was concentrated. Purification by column chromatography afforded 12-(benzyloxy)dodecan-6-ol (2.57 g) as a colorless liquid.

[1038] Step 4: Synthesis of 12-(benzyloxy)dodec-6-ylbutyl ester

[1039] Reaction formula:

[1040]

[1041] Material ratio:

[1042]

[1043] Operation process:

[1044] 12-(Benzyloxy)dodecan-6-ol (2.57 g, 8.79 mmol) and n-butyric acid (929 mg, 10.5 mmol) were dissolved in dichloromethane (25.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.53 g, 13.1 mmol) and 4-dimethylaminopyridine (107 mg, 878 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave 12-(Benzyloxy)dodecan-6-ylbutyl ester (2.25 g) as a colorless liquid.

[1045] Step 5: Synthesis of 12-hydroxydodecane-6-yl butyl ester

[1046] Reaction formula:

[1047]

[1048] Material ratio:

[1049]

[1050]

[1051] Operation process:

[1052] 12-(Benzyloxy)dodec-6-ylbutyl ester (2.25 g, 6.21 mmol) was dissolved in methanol (50.0 mL), palladium carbon (1.32 g, 1.24 mmol) was added, and the mixture was reacted at 35°C, 35 Psi under a hydrogen atmosphere for 12 hours. The reaction was completed after TLC monitoring. The mixture was filtered and concentrated to give a colorless liquid 12-hydroxydodec-6-ylbutyl ester (1.50 g).

[1053] Step 6: Synthesis of 1-[(5-bromovaleryl)oxy]dodec-4-ylheptyl ester

[1054] Reaction formula:

[1055]

[1056] Material ratio:

[1057]

[1058] Operation process:

[1059] 12-Hydroxydodec-6-ylbutyl ester (1.50 g, 5.51 mmol) and 5-bromovaleric acid (1.20 g, 6.61 mmol) were dissolved in dichloromethane (15.0 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.58 g, 8.26 mmol) and 4-dimethylaminopyridine (67.7 mg, 550 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave 1-[(5-bromovaleryl)oxy]dodec-4-ylheptyl ester (2.20 g) as a colorless liquid.

[1060] Step 7: Synthesis of heptadecane-9-yl 8-[(5-{[7-(butyloxo)dodecyl]oxy}-5-oxypentylidene)(2-hydroxyethyl)amino]octyl ester

[1061] Reaction formula:

[1062]

[1063] Material ratio:

[1064]

[1065]

[1066] Operation process:

[1067] 1-[(5-bromovaleroyl)oxy]dodec-4-ylheptyl ester (1.08 g, 2.49 mmol) was dissolved in acetonitrile (10.0 mL). Heptadec-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (1.00 g, 2.26 mmol), potassium carbonate (1.10 g, 7.92 mmol), potassium iodide (450 mg, 2.72 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC and concentrated. Purification by column chromatography gave heptadec-9-yl 8-[(5-{[7-(butyloxo)dodecyl]oxy}-5-oxypentylene)(2-hydroxyethyl)amino]octyl ester (1.50 g), a colorless liquid.

[1068] 1H NMR(400MHz,CHLOROFORM-d)δ=5.03-4.76(m,2H),4.20-3.92(m,2H),3.70-3.43(m,2H) ),2.70-2.58(m,2H),2.57-2.43(m,4H),2.40-2.19(m,6H),1.69-1.58(m,8H),1.52(br s,12H),1.38-1.19(m,42H),1.06-0.70(m,12H)ppm.

[1069] LCMS:RT=2.037,m / z 796.5[M+H] + .

[1070] Preparation Example 30 Preparation of Compound 58

[1071] Step 1: Synthesis of 2-hydroxydodecyl 6-bromohexyl ester

[1072] Reaction formula:

[1073]

[1074] Material ratio:

[1075] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Decyloxapropane 184 1.0 eq 4.00g 21.7mmol 6-Bromohexanoic acid 193 1.05eq 4.44g 22.7mmol Ferric chloride 162 0.025eq 95.2mg 0.587mmol Pyridine 79 0.025eq 46.4mg 0.587mmol

[1076] Operation process:

[1077] To a reaction flask, 2-decyloxirane (4.00 g, 21.7 mmol), 6-bromohexanoic acid (4.44 g, 22.7 mmol), ferric chloride (88.0 mg, 542 μmol), and pyridine (42.9 mg, 542 μmol) were added sequentially. The mixture was then reacted at 30°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction solution was concentrated and purified by column chromatography to yield 2-hydroxydodecyl 6-bromohexyl ester (5.60 g), a colorless liquid.

[1078] Step 2: Synthesis of 2-[(3-ethylpentanoyl)oxy]dodecyl 6-bromohexyl ester

[1079] Reaction formula:

[1080]

[1081] Material ratio:

[1082]

[1083]

[1084] Operation process:

[1085] 2-Hydroxydodecyl 6-bromohexyl ester (3.00 g, 7.91 mmol) was dissolved in dichloromethane (30.0 mL). 3-Ethylpentanoic acid (1.03 g, 7.91 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27 g, 11.8 mmol), and 4-dimethylaminopyridine (96.6 mg, 790 μmol) were added sequentially. The mixture was reacted under nitrogen for 12 hours at 25°C. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain 2-[(3-ethylpentanoyl)oxy]dodecyl 6-bromohexyl ester (2.47 g), a colorless liquid.

[1086] Step 3: Synthesis of 2-[(3-ethylpentanoyl)oxy]dodecyl 6-{[6-({2-[(3-ethylpentanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl](2-hydroxyethyl)amino}hexyl ester

[1087] Reaction formula:

[1088]

[1089] Material ratio:

[1090]

[1091] Operation process:

[1092] 2-[(3-ethylpentanoyl)oxy]dodecyl 6-bromohexyl ester (1.00 g, 2.03 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (59.6 mg, 976 μmol), potassium carbonate (984 mg, 7.12 mmol), potassium iodide (405 mg, 2.44 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-[(3-ethylpentanoyl)oxy]dodecyl 6-{[6-({2-[(3-ethylpentanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl](2-hydroxyethyl)amino}hexyl ester (520 mg) as a colorless liquid.

[1093] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.10 (dq, J1 = 3.4, J2 = 6.6Hz, 2H), 4.24 (dd, J1 = 3.4, J2 = 11.8Hz, 2H), 4.02 (dd, J1 = 7.0, J2 = 11.8Hz, 2H), 3.59 (br s,2H),2.65(br s,2H),2.53(br s,4H),2.31(t,J=7.4Hz,4H),2.25(d,J=7.0Hz,4H),1.75(td,J1=6.4,J2=1 3.2Hz,2H),1.66-1.46(m,12H),1.38-1.20(m,44H),0.97-0.81(m,18H)ppm.

[1094] LCMS: RT=2.069, m / z 882.6[M+H] + .

[1095] Preparation Example 31 Preparation of Compound 59

[1096] Step 1: Preparation of 59-1

[1097] Reaction formula:

[1098]

[1099] Material ratio:

[1100] Material Name Molecular weight Feed ratio Feeding amount mmol glycerin 92 1eq 9.2g 100 Imidazole 68 2.5eq 17g 250 tert-Butyldimethylsilyl chloride 150.72 2.1eq 31.7g 210 Tetrahydrofuran - - 380mL -

[1101] Operation process:

[1102] Glycerol, imidazole, and tetrahydrofuran (200 mL) were added to the reaction flask, cooled to 0-5°C, and a solution of tert-butyldimethylsilyl chloride in tetrahydrofuran (180 mL) was added dropwise. The reaction was stirred at room temperature for 16 hours. TLC (PE:EA = 10:1) revealed an Rf value of 0.3 for the product. 500 mL of water was added to the reaction solution, followed by extraction with 400 mL of ethyl acetate. The organic phase was dried, concentrated, and purified by column chromatography to yield 28 g of a colorless oil.

[1103] Step 2: Preparation of 10-2

[1104] Reaction formula:

[1105]

[1106] Material ratio:

[1107] Material Name Molecular weight Feed ratio Feeding amount mmol 59-1 320.62 1 eq 12.8g 40 8-Bromooctanoic acid 223.11 1.1 eq 9.8g 44 EDCI 192 2eq 15.4g 80 DMAP 122 0.2eq 1g 8 dichloromethane - - 200mL -

[1108] Operation process:

[1109] 59-1, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) revealed an Rf value of 0.5 for the product. The reaction mixture was washed once with 200 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 13.2 g of a colorless oil.

[1110] Step 3: Preparation of 10-3

[1111] Reaction formula:

[1112]

[1113] Material ratio:

[1114]

[1115] Operation process:

[1116] Add 59-2 and tetrahydrofuran to the reaction flask, followed by a 1M solution of tetrabutylammonium fluoride in tetrahydrofuran. Stir the reaction at room temperature for 16 hours. TLC (DCM:MeOH = 10:1) reveals an Rf value of 0.5 for the product. Extract the reaction solution with 400 mL of water and 400 mL of ethyl acetate. The organic phase is dried, concentrated, and purified by column chromatography to yield 6.5 g of a colorless oil.

[1117] Step 4: Preparation of 10-4

[1118] Reaction formula:

[1119]

[1120] Material ratio:

[1121] Material Name Molecular weight Feed ratio Feeding amount mmol 59-3 297.19 1eq 6g 20 bitter 144.21 2.2 eq 6.3g 44 EDCI 192 3eq 11.5g 60 DMAP 122 0.2 eq 488mg 4 DCM - - 100mL -

[1122] Operation process:

[1123] To the reaction flask, 59-3, octanoic acid, EDCI, DMAP, and DCM were added and stirred at room temperature for 16 h. TLC (PE:EA = 20:1) revealed an Rf value of 0.6 for the product. The reaction mixture was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 8.2 g of a colorless oil.

[1124] Step 5: Preparation of compound 10

[1125] Reaction formula:

[1126]

[1127] Material ratio:

[1128] Material Name Molecular weight Feed ratio Feeding amount mmol 59-4 549.59 1.1eq 4.7g 8.55 SM102-3 329.53 1eq 2.56g 7.77 potassium carbonate 138 1.5 eq 1.6g 11.6 Potassium iodide 166 1.5 eq 1.9g 11.6 Acetonitrile - - 40mL -

[1129] Operation process:

[1130] 10-4, SM102-3, K2CO3, KI, and acetonitrile were added to a reaction flask and stirred at 65°C for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was concentrated, diluted with 100 mL of ethyl acetate, and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.8 g of a colorless oil.

[1131] 1 H NMR (600MHz, Chloroform-d) δ5.26 (ddd, J=10.2, 6.0, 4.3Hz, 1H), 4.28 (ddd, J=11.9, 4. 3,1.7Hz,2H),4.14(dd,J=11.9,6.0Hz,2H),4.05(t,J=6.8Hz,2H),3.53(t,J=5.3Hz,2H ),2.59(t,J=5.3Hz,2H),2.46(q,J=7.6Hz,4H),2.33–2.27(m,8H),1.61(q,J=8.7,8.2H z,10H),1.50–1.40(m,4H),1.28(ddd,J=22.6,11.5,4.6Hz,41H),0.87(t,J=6.9Hz,9H).

[1132] MS (ES+) m / z): 798.7 (M+H) + .

[1133] Preparation Example 32 Preparation of Compound 60

[1134] Step 1: Preparation of 60-1

[1135] Reaction formula:

[1136]

[1137] Material ratio:

[1138] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Hydroxymethyl-1,3-propanediol 106.12 1eq 3g 28.3 Octanoyl chloride 162.66 1.95eq 9g 55.12 Pyridine 79 4 9g 113 DMAP 122 0.4 1.4g 11 DCM - - 100mL -

[1139] Operation process:

[1140] 2-Hydroxymethyl-1,3-propanediol, octanoyl chloride, pyridine, DMAP, and DCM were added to a reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 4:1) revealed an Rf value of 0.4 for the product. The reaction mixture was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 8 g of the product.

[1141] Step 2: Preparation of 60-2

[1142] Reaction formula:

[1143]

[1144] Material ratio:

[1145] Material Name Molecular weight Feed ratio Feeding amount mmol 60-1 358.52 1eq 6g 16.7 7-Bromoheptanoic acid 209.08 1.1eq 3.85g 18.4 EDCI 192 1.5eq 4.8g 25 DMAP 122 0.15 eq 300mg 2.5 DCM - - 100mL -

[1146] Operation process:

[1147] 60-1, 7-bromoheptanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and allowed to react at room temperature for 16 h. The product had an Rf value of 0.5 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 200 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 5 g of an oil.

[1148] Step 3: Preparation of compound 60

[1149] Reaction formula:

[1150]

[1151] Material ratio:

[1152] Material Name Molecular weight Feed ratio Feeding amount mmol 60-2 549.59 1.1eq 2.2g 4 SM102-3 329.53 1 eq 1.2g 3.7 potassium carbonate 138 1.5 eq 770mg 5.6 Potassium iodide 166 1 eq 660mg 4 Acetonitrile - - 50mL -

[1153] Operation process:

[1154] 60-2, SM102-3, potassium carbonate, potassium iodide, and acetonitrile were added to the reaction flask and stirred at 65°C for 16 hours. The product had an rR value of 0.6, as determined by TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate and washed with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 1.2 g of an oil.

[1155] 1 H NMR(600MHz,Chloroform-d)δ4.12(d,J=6.0Hz,6H),4.05(t,J=6.8Hz,2H),3.55(t,J=5.3Hz,2H),2.60(t,J=5.4Hz,2H),2.48(dq,J=8.2,4.7 Hz,4H),2.39(p,J=6.0Hz,1H),2.30(tt,J=7.3,3.3Hz,8H),1.65–1.58(m,10H),1.51–1.43(m,4H),1.34–1.24(m,40H),0.88(t,J=7.0Hz,9H).

[1156] MS (ES+) m / z): 798.7 (M+H) + .

[1157] Preparation Example 33 Preparation of Compound 66

[1158] Step 1: Preparation of 66-1

[1159] Reaction formula:

[1160]

[1161] Material ratio:

[1162] Material Name Molecular weight Feed ratio Feeding amount mmol 5-Bromopentanoic acid 181 1 eq 18.1g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mg 2.5

[1163] Operation process:

[1164] 5-Bromopentanoic acid, 1,2-epoxydodecane, pyridine, and ferric chloride were added to the reaction flask and reacted at room temperature for 16 hours. The product had an Rf value of 0.4 and was purified by TLC (PE:EA = 4:1) column chromatography to obtain 18 g of an oil.

[1165] Step 2: Preparation of 66-2

[1166] Reaction formula:

[1167]

[1168] Material ratio:

[1169] Material Name Molecular weight Feed ratio Feeding amount mmol 66-1 365 1eq 11g 30 bitter 144 1 eq 4.4g 30 EDCI 192 1.5eq 5.8g 45 DMAP 122 0.15 eq 400mg 4.5 DCM - - 100mL -

[1170] Operation process:

[1171] Add 66-1, octanoic acid, EDCI, DMAP, and DCM to a reaction flask and react at room temperature for 2 h. The product has an Rf value of 0.5 according to TLC (PE:EA = 10:1). Wash the reaction solution once with 100 mL of water, dry the organic phase over anhydrous sodium sulfate, concentrate, and purify by column chromatography to yield 8 g of an oil.

[1172] Step 3: Preparation of 66-3

[1173] Reaction formula:

[1174]

[1175] Material ratio:

[1176]

[1177]

[1178] Operation process:

[1179] 8-Bromooctanoic acid, 9-heptadecanol, EDCI, DMAP, and DCM were added to a reaction flask and allowed to react at room temperature for 2 h. The product exhibited an Rf value of 0.5 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 10.8 g of an oil.

[1180] Step 4: Preparation of 66-4

[1181] Reaction formula:

[1182]

[1183] Material ratio:

[1184] Material Name Molecular weight Feed ratio Feeding amount mmol 66-3 461 1eq 9.2g 20 Ethanolamine 61 1 eq 6.1g 100 Acetonitrile - - 80mL -

[1185] Operation process:

[1186] Add 66-3, ethanolamine, and acetonitrile to the reaction flask and stir at room temperature for 16 hours. TLC (DCM:MeOH = 10:1) shows an Rf value of 0.3 for the product. Dilute the reaction solution with 200 mL of ethyl acetate and wash twice with 200 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 5.6 g of a colorless oil.

[1187] Step 5: Preparation of compound 66

[1188] Reaction formula:

[1189]

[1190] Material ratio:

[1191] Material Name Molecular weight Feed ratio Feeding amount mmol 66-4 441 1eq 2.2g 5 66-2 491 1.1 eq 2.7g 5.5 potassium carbonate 138 1.5eq 1g 7.5 Potassium iodide 166 0.5 eq 420mg 2.5 Acetonitrile - - 50mL -

[1192] Operation process:

[1193] To a reaction flask, add 66-2, 66-4, potassium carbonate, potassium iodide, and acetonitrile and react at 65°C for 16 h. The product's rR value is 0.6, as determined by TLC (DCM:MeOH = 20:1). The reaction mixture is diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.8 g of an oil.

[1194] 1H NMR(600MHz,Chloroform-d)δ5.10–5.05(m,1H),4.86(p,J=6.2Hz,1H),4.23(dd,J=11 .8,3.3Hz,1H),4.01(dd,J=11.8,6.9Hz,1H),3.55(t,J=5.3Hz,2H),2.60(t,J=5.3Hz, 2H),2.48(dt,J=16.5,7.5Hz,4H),2.29(dq,J=22.5,7.4Hz,6H),1.65–1.53(m,8H),1. 47(dq,J=33.8,8.0,7.4Hz,8H),1.32–1.23(m,55H),0.87(td,J=7.3,6.8,1.9Hz,12H).

[1195] MS (ES+) m / z): 853.7 (M+H) + .

[1196] Preparation Example 34 Preparation of Compound 33

[1197] Step 1: Preparation of compound 33-1

[1198] Reaction formula:

[1199]

[1200] Material ratio:

[1201] Material Name Molecular weight Feed ratio Feeding amount mmol 6-1 379.38 1eq 6.9g 18.2 n-heptanoic acid 130 1eq 2.37g 18.2 EDCI 192 1.5eq 5.2g 27 DMAP 122 0.15 eq 330mg 2.7 DCM - - 100mL -

[1202] Operation process:

[1203] To the reaction flask, 6-1, n-heptanoic acid, EDCI, DMAP, and DCM were added and allowed to react at room temperature for 16 h. The product had an Rf value of 0.5 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 5.6 g of an oil.

[1204] Step 2: Preparation of compound 33

[1205] Reaction formula:

[1206]

[1207] Material ratio:

[1208]

[1209]

[1210] Operation process:

[1211] To the reaction flask, 33-1, ethanolamine, potassium carbonate, potassium iodide, and acetonitrile were added and stirred at 65°C for 16 h. The product exhibited an rR value of 0.6, as determined by TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 1 g of an oil.

[1212] 1 H NMR (400MHz, Chloroform-d) δ5.08 (qd, J=6.7, 3.3Hz, 2H), 4.22 (dd, J=11.8, 3.3 Hz,2H),4.02(dd,J=11.8,6.8Hz,2H),3.54(t,J=5.3Hz,2H),2.58(t,J=5.3Hz,2 H),2.46(t,J=7.5Hz,4H),2.34–2.26(m,8H),1.61(tdd,J=16.0,13.8,7.9,4.6H z,12H),1.46(p,J=7.6Hz,4H),1.34–1.21(m,48H),0.88(td,J=6.9,2.8Hz,12H).

[1213] MS (ES+) m / z): 882.6 (M+H) + .

[1214] Preparation Example 35 Preparation of Compound 35

[1215] Step 1: Synthesis of 2-hydroxydodecyl 6-bromohexyl ester

[1216] Reaction formula:

[1217]

[1218] Material ratio:

[1219] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Decyloxapropane 184 1.0 eq 4.00g 21.7 6-Bromohexanoic acid 195 1.05eq 4.44g 22.7 Pyridine 79.1 0.025eq 42.9mg 0.542 Ferric chloride 162 0.025eq 88.0mg 0.542

[1220] Operation process:

[1221] Dissolve 6-bromohexanoic acid (4.44 g, 22.7 mmol) in 2-decyloxirane (4.00 g, 21.7 mmol), add pyridine (42.9 mg, 0.542 mmol) and ferric chloride (88.0 mg, 0.542 mmol). Under nitrogen, react at 30°C for 12 hours. TLC monitoring indicates the formation of new spots. The organic phase is concentrated. Column chromatography yields the crude product as a white solid, 2-hydroxydodecyl 6-bromohexyl ester (5.50 g, 66.8% yield).

[1222] Step 2: Synthesis of 1-[(6-bromohexanoyl)oxy]dodec-2-yl 3-ethylheptyl ester

[1223] Reaction formula:

[1224]

[1225] Material ratio:

[1226]

[1227]

[1228] Operation process:

[1229] 2-Hydroxydodecyl 6-bromohexyl ester (1.20 g, 3.16 mmol) and 3-ethylheptanoic acid (500 mg, 3.16 mmol) were dissolved in tetrahydrofuran (12 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (909 mg, 4.74 mmol) and 4-dimethylaminopyridine (77.3 mg, 0.632 mmol) were added sequentially. The mixture was reacted at 40°C under nitrogen for 16 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to yield 1-[(6-bromohexanoyl)oxy]dodec-2-yl 3-ethylheptyl ester (1.1 g, 66.9% yield) as a colorless liquid.

[1230] Step 3: Synthesis of 1-[(6-{[6-({2-[(3-ethylheptanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl](2-hydroxyethyl)amino}hexanoyl)oxy]dodec-2-yl 3-ethylheptyl ester

[1231] Reaction formula:

[1232]

[1233] Material ratio:

[1234]

[1235] Operation process:

[1236] 1-[(6-bromohexanoyl)oxy]dodec-2-yl 3-ethylheptyl ester (1.10 g, 2.12 mmol) was dissolved in acetonitrile (6 mL), and 2-aminoethane-1-ol (62.0 mg, 1.02 mmol), potassium carbonate (877 mg, 6.35 mmol), potassium iodide (421 mg, 2.54 mmol), and tetrahydrofuran (6 mL) were added in sequence. Finally, the mixture was reacted at 70° C. under nitrogen protection for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a yellow liquid 1-[(6-{[6-({2-[(3-ethylheptanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl](2-hydroxyethyl)amino}hexanoyl)oxy]dodec-2-yl 3-ethylheptyl ester (320 mg, 13.6% yield).

[1237] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.15-5.05 (m, 2H), 4.23 (dd, J = 3.2, 12.0Hz, 2H), 4.02 (dd, J = 6.8, 12.0Hz, 2H), 3.53 (br t,J=5.2Hz,2H),2.62-2.54(m,2H),2.46(brt,J=7.2Hz,4H),2.31(t,J=7.6Hz,4H),2.25(d,J=6.8Hz, 4H),1.86-1.74(m,2H),1.69-1.60(m,9H),1.50-1.43(m,4H),1.40-1.20(m,54H),0.95-0.81(m,18H).

[1238] LCMS:RT=2.297,m / z 994.9[M+H] + .

[1239] Preparation Example 36 Preparation of Compound 36

[1240] Step 1: Synthesis of 3-ethylheptanenitrile

[1241] Reaction formula:

[1242]

[1243] Material ratio:

[1244] Material Name Molecular weight Feed ratio Feeding amount mmol 3-(Bromomethyl)heptane 192 1.0 eq 19.0g 98.3 Potassium cyanide 64.9 1.5eq 9.61g 147 Potassium iodide 165 0.10eq 1.63g 9.84 N,N-dimethylformamide - - 190mL -

[1245] Operation process:

[1246] Potassium cyanide (9.61 g, 147 mmol) was dissolved in N,N-dimethylformamide (90 mL) and reacted at 25°C for 0.5 hours. 3-(Bromomethyl)heptane (19.0 g, 98.3 mmol) was dissolved in N,N-dimethylformamide (100 mL) at 25°C and added dropwise to the reaction solution. Potassium iodide (1.63 g, 9.84 mmol) was added to the reaction solution, and the reaction was continued at 100°C under nitrogen for 16 hours. TLC monitoring revealed the formation of new spots. 1000 mL of water was added, and the mixture was extracted with petroleum ether twice, 500 mL each time. The organic phase was concentrated and purified by column chromatography to obtain 3-ethylheptanenitrile (1.67 g, 12.1% yield), a colorless liquid.

[1247] Step 2: Synthesis of 3-ethylheptylamide

[1248] Reaction formula:

[1249]

[1250] Material ratio:

[1251] Material Name Molecular weight Feed ratio Feeding amount mmol 3-Ethylheptanenitrile 139 1.0 eq 1.67g 11.9 potassium hydroxide 55.9 3.0eq 2.02g 35.9 ethanol - - 16mL -

[1252] Operation process:

[1253] Dissolve 3-ethylheptanenitrile (1.67 g, 11.9 mmol) in ethanol (16 mL), add potassium hydroxide (2.02 g, 2.5 M, 35.9 mmol), and react at 110°C under nitrogen for 24 hours. TLC monitoring indicates the formation of new spots. Concentrate to dryness, add 10 mL of water, and extract with dichloromethane twice, each time using 50 mL. The organic phase is concentrated. Column chromatography yields the crude product as a white solid, 3-ethylheptaneamide (680 mg, 36.0% yield).

[1254] Step 3: Synthesis of 3-ethylheptanoic acid

[1255] Reaction formula:

[1256]

[1257] Material ratio:

[1258]

[1259]

[1260] Operation process:

[1261] Dissolve 3-ethylheptylamide (1.30 g, 8.27 mmol) in dioxane (13 mL), add sulfuric acid (3.83 g, 3.0 M, 39.0 mmol), and react at 80°C under nitrogen for 16 hours. TLC monitoring indicates the formation of new spots. Concentrate to dryness, extract with dichloromethane twice, 50 mL each time, and concentrate the organic phase. This yields 3-ethylheptyl acid (1.31 g, 88.6% yield) as a white solid.

[1262] Step 4: Synthesis of 1-(benzyloxy)dodec-2-yl 3-ethylheptyl ester

[1263] Reaction formula:

[1264]

[1265] Material ratio:

[1266]

[1267] Operation process:

[1268] 1-(Benzyloxy)dodecan-2-ol (2.22 g, 7.58 mmol) and 3-ethylheptanoic acid (1.00 g, 6.32 mmol) were dissolved in dichloromethane (10 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.82 g, 9.48 mmol) and 4-dimethylaminopyridine (77.2 mg, 0.631 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodecan-2-yl 3-ethylheptyl ester (2.15 g, 78.6% yield) as a colorless liquid.

[1269] Step 5: Synthesis of 1-hydroxydodec-2-yl 3-ethylheptyl ester

[1270] Reaction formula:

[1271]

[1272] Material ratio:

[1273]

[1274] Operation process:

[1275] Dissolve 1-(Benzyloxy)dodec-2-yl 3-ethylheptyl ester (2.15 g, 4.97 mmol) in methanol (21 mL), add palladium on carbon (1.06 g, 0.993 mmol), and react under hydrogen at 35°C, 40 psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1-hydroxydodec-2-yl 3-ethylheptyl ester (1.54 g, 90.4% yield) as a colorless liquid.

[1276] Step 6: Synthesis of 1-[(4-bromobutyryl)oxy]dodec-2-yl 3-ethylheptyl ester

[1277] Reaction formula:

[1278]

[1279] Material ratio:

[1280]

[1281] Operation process:

[1282] 1-Hydroxydodec-2-yl 3-ethylheptyl ester (750 mg, 2.19 mmol) and 4-bromobutyric acid (402 mg, 2.41 mmol) were dissolved in dichloromethane (8 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (629 mg, 3.28 mmol) and 4-dimethylaminopyridine (26.7 mg, 0.218 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to afford 1-[(4-bromobutyryl)oxy]dodec-2-yl 3-ethylheptyl ester (850 mg, 78.9% yield) as a colorless liquid.

[1283] Step 7: Synthesis of 1-[(4-{[4-({2-[(3-ethylheptanoyl)oxy]dodecyl}oxy)-4-oxyylidenebutyl](2-hydroxyethyl)amino}butyryl)oxy]dodec-2-yl 3-ethylheptyl ester

[1284] Reaction formula:

[1285]

[1286] Material ratio:

[1287]

[1288] Operation process:

[1289] 1-[(4-bromobutyryl)oxy]dodec-2-yl 3-ethylheptyl ester (750 mg, 1.53 mmol) was dissolved in acetonitrile (7 mL), and 2-aminoethane-1-ol (41.9 mg, 0.686 mmol), potassium carbonate (632 mg, 4.58 mmol), potassium iodide (379 mg, 2.29 mmol), and tetrahydrofuran (3 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-[(4-{[4-({2-[(3-ethylheptanoyl)oxy]dodecyl}oxy)-4-oxylidenebutyl](2-hydroxyethyl)amino}butyryl)oxy]dodec-2-yl 3-ethylheptyl ester (340 mg, 24.0% yield) as a bright yellow liquid.

[1290] 1 H NMR (400MHz, CHLOROFORM-d) δ=5.20-5.00(m,2H),4.24(dd,J=3.2,12.0Hz,2H),4.01(dd,J=6.8,12.0Hz,2H),3.62-3.50(m,2H),2.61(br s,2H),2.51(br t,J=6.8Hz,4H),2.32(t,J=7.2Hz,4H),2.25(d,J=6.8Hz,4H),1.84-1.74(m,6H),1.59(br s,13H),1.33-1.26(m,40H),0.95-0.84(m,18H).

[1291] LCMS:RT=2.213,m / z 882.7[M+H] + .

[1292] Preparation Example 37 Preparation of Compound 37

[1293] Step 1: Synthesis of 2-[(3-ethylheptanoyl)oxy]dodecyl 8-bromooctyl ester

[1294] Reaction formula:

[1295]

[1296] Material ratio:

[1297]

[1298] Operation process:

[1299] 1-Hydroxydodec-2-yl 3-ethylheptyl ester (750 mg, 2.19 mmol) and 8-bromooctanoic acid (537 mg, 2.41 mmol) were dissolved in dichloromethane (8 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (629 mg, 3.28 mmol) and 4-dimethylaminopyridine (26.7 mg, 0.218 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to yield 2-[(3-ethylheptyl)oxy]dodecyl 8-bromooctyl ester (1.15 g, 95.9% yield) as a colorless liquid.

[1300] Step 2: Synthesis of 2-[(3-ethylheptanoyl)oxy]dodecyl 8-{[8-({2-[(3-ethylheptanoyl)oxy]dodecyl}oxy)-8-oxyylideneoctyl](2-hydroxyethyl)amino}octyl ester

[1301] Reaction formula:

[1302]

[1303] Material ratio:

[1304]

[1305] Operation process:

[1306] 2-[(3-ethylheptanoyl)oxy]dodecyl 8-bromooctyl ester (1.00 g, 1.83 mmol) was dissolved in acetonitrile (7 mL), and 2-aminoethane-1-ol (55.7 mg, 0.913 mmol), potassium carbonate (757 mg, 5.48 mmol), potassium iodide (454 mg, 2.74 mmol), and tetrahydrofuran (3 mL) were added in sequence. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a yellow liquid, 2-[(3-ethylheptanoyl)oxy]dodecyl 8-{[8-({2-[(3-ethylheptanoyl)oxy]dodecyl}oxy)-8-oxyideneoctyl](2-hydroxyethyl)amino}octyl ester (656 mg, 34.6% yield).

[1307] 1H NMR (400MHz, CHLOROFORM-d) δ=5.16-5.03(m,2H),4.23(dd,J=3.2,12.0Hz,2H),4.02(dd,J=6.8,11.6Hz,2H),3.64-3.54(m,2H),2.65(br s,2H),2.52(br s,3H),2.30(t,J=7.6Hz,4H),2.25(d,J=6.8Hz,4H),1.85-1.74(m,3H),1.61(br s,8H),1.52-1.42(m,6H),1.37-1.22(m,60H),0.96-0.82(m,18H).

[1308] LCMS:RT=2.297,m / z 994.9[M+H] + .

[1309] Preparation Example 38 Preparation of Compound 47

[1310] Step 1: Synthesis of 1-[(6-bromohexanoyl)oxy]dodec-4-ylheptyl ester

[1311] Reaction formula:

[1312]

[1313] Material ratio:

[1314]

[1315]

[1316] Operation process:

[1317] 1-Hydroxydodec-4-ylheptyl ester (1.60 g, 5.09 mmol) and 6-bromohexanoic acid (1.19 g, 6.10 mmol) were dissolved in dichloromethane (16.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.46 g, 7.63 mmol) and 4-dimethylaminopyridine (62.1 mg, 508 μmol) were added sequentially. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After addition of 100 mL of water, the mixture was extracted with dichloromethane twice, each time using 100 mL. The organic phase was concentrated. Purification by column chromatography afforded 1-[(6-bromohexanoyl)oxy]dodec-4-ylheptyl ester (1.70 g, 67.9% yield) as a bright yellow liquid.

[1318] Step 2: Synthesis of 1-({6-[(6-{[4-(heptanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexanoyl}oxy)dodec-4-ylheptyl ester

[1319] Reaction formula:

[1320]

[1321] Material ratio:

[1322]

[1323] Operation process:

[1324] 1-[(6-bromohexanoyl)oxy]dodec-4-ylheptyl ester (1.00 g, 2.03 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (59.6 mg, 976 μmol), potassium carbonate (984 mg, 7.12 mmol), potassium iodide (405 mg, 2.44 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({6-[(6-{[4-(heptanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexanoyl}oxy)dodec-4-ylheptyl ester (608 mg, 32.7% yield) as a colorless liquid.

[1325] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.90 (quin, J = 6.0Hz, 2H), 4.17-3.93 (m, 4H), 3.56 (t, J = 5.2Hz, 2H), 2.62 (br t, J = 5.2Hz, 2H), 2.50 (br t,J=7.4Hz,4H),2.30(dt,J1=4.8,J2=7.6Hz,8H),1.67-1.47(m,24H),1.34-1.16(m,40H),0.98-0.76(m,12H)ppm.

[1326] LCMS: RT = 2.059, m / z 882 [M+H] + .

[1327] Preparation Example 39 Preparation of Compound 49

[1328] Step 1: Synthesis of 1-(benzyloxy)dodecan-2-ol

[1329] Reaction formula:

[1330]

[1331] Material ratio:

[1332] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Benzyloxy)acetaldehyde 150 1.0eq 5.0g 33.29 Bromo(decyl)magnesium 244 1.2eq 9.81g 39.95 Tetrahydrofuran - - 50mL -

[1333] Operation process:

[1334] 2-(Benzyloxy)acetaldehyde (5.0 g, 33.29 mmol) was dissolved in tetrahydrofuran (50 mL). Decylmagnesium bromide (9.81 g, 0.5 M, 79.91 mL, 19.06 mmol) was added at 0°C. The mixture was reacted at 25°C under nitrogen for 12 hours. After completion of the reaction, 50 mL of aqueous ammonium chloride was added. The mixture was extracted with ethyl acetate twice, using 200 mL each extraction. The organic phase was concentrated and purified by column chromatography to afford 1-(benzyloxy)dodecan-2-ol (7.5 g, 77.0% yield), a colorless liquid.

[1335] Step 2: Synthesis of 1-(benzyloxy)dodec-2-ylpentyl ester

[1336] Reaction formula:

[1337]

[1338] Material ratio:

[1339]

[1340] Operation process:

[1341] 1-(Benzyloxy)dodecan-2-ol (4.0 g, 13.6 mmol) and valeric acid (2.10 g, 20.5 mmol) were dissolved in dichloromethane (40 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.93 g, 20.5 mmol) and 4-dimethylaminopyridine (167 mg, 1.37 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodecan-2-ylpentyl ester (3.8 g, 73.7% yield) as a colorless liquid.

[1342] Step 3: Synthesis of 1-hydroxydodec-2-ylpentyl ester

[1343] Reaction formula:

[1344]

[1345] Material ratio:

[1346] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodec-2-ylpentyl ester 376 1.0 eq 3.80g 10.0 Palladium on carbon 105.9 0.2eq 2.15g 2.02 Methanol - - 40mL -

[1347] Operation process:

[1348] Dissolve 1-(Benzyloxy)dodec-2-ylpentyl ester (3.80 g, 10.0 mmol) in methanol (40 mL), add palladium on carbon (2.15 g, 2.02 mmol), and react under hydrogen at 35°C, 40 psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1-hydroxydodec-2-ylpentyl ester (2.00 g, 69.1% yield) as a colorless liquid.

[1349] Step 4: Synthesis of 2-(pentanoyloxy)dodecyl 5-bromopentyl ester

[1350] Reaction formula:

[1351]

[1352] Material ratio:

[1353]

[1354] Operation process:

[1355] 1-Hydroxydodec-2-ylpentyl ester (1.00 g, 3.49 mmol) and 5-bromovaleric acid (758 mg, 4.19 mmol) were dissolved in dichloromethane (80 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.00 g, 5.24 mmol) and 4-dimethylaminopyridine (42.6 mg, 0.349 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-(pentanoyloxy)dodecyl 5-bromopentyl ester (1.40 g, 89.2% yield) as a colorless liquid.

[1356] Step 5: Synthesis of heptadecane-9-yl 8-[(2-hydroxyethyl)(5-oxyylidene-5-{[2-(pentanoyloxy)dodecyl]oxy}pentyl)amino]octyl ester

[1357] Reaction formula:

[1358]

[1359] Material ratio:

[1360]

[1361] Operation process:

[1362] 2-(Valeryloxy)dodecyl 5-bromopentyl ester (1.12 g, 2.49 mmol) was dissolved in acetonitrile (8 mL), and heptadecan-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (1.00 g, 2.26 mmol), potassium carbonate (1.10 g, 7.92 mmol), potassium iodide (450 mg, 2.72 mmol), and tetrahydrofuran (2 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadecan-9-yl 8-[(2-hydroxyethyl)(5-oxyidene-5-{[2-(pentanoyloxy)dodecyl]oxy}pentyl)amino]octyl ester (1.00 g, 50.2% yield) as a yellow liquid.

[1363] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.11 (dq, J = 3.2, 6.8Hz, 1H), 4.88 (quin, J = 6.4Hz, 1H), 4.26 (dd, J = 3.2, 12.0Hz, 1H), 4.04 (dd, J = 6.8, 12.0Hz, 1H), 3.58 (br t,J=5.2Hz,2H),2.63(br t,J=5.2Hz,2H),2.56-2.47(m,4H),2.32(dt,J=7.2,13.2Hz,6H),1.63(quin ,J=7.6Hz,7H),1.59-1.44(m,10H),1.42-1.23(m,48H),0.99-0.85(m,12H).

[1364] LCMS:RT=2.146,m / z 810.7[M+H] + .

[1365] Preparation Example 40 Preparation of Compound 50

[1366] Step 1: Synthesis of 2-(pentanoyloxy)dodecyl 6-bromohexyl ester

[1367] Reaction formula:

[1368]

[1369] Material ratio:

[1370]

[1371] Operation process:

[1372] 1-Hydroxydodec-2-ylpentyl ester (1.00 g, 3.49 mmol) and 6-bromohexanoic acid (817 mg, 4.19 mmol) were dissolved in dichloromethane (20 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.00 g, 5.24 mmol) and 4-dimethylaminopyridine (42.6 mg, 0.349 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-(pentanoyloxy)dodecyl 6-bromohexyl ester (1.5 g, 92.7% yield) as a colorless liquid.

[1373] Step 2: Synthesis of 2-(pentanoyloxy)dodecyl 6-[(2-hydroxyethyl)(6-oxyylidene-6-{[2-(pentanoyloxy)dodecyl]oxy}hexyl)amino]hexyl ester

[1374] Reaction formula:

[1375]

[1376] Material ratio:

[1377]

[1378] Operation process:

[1379] 2-(Valeryloxy)dodecyl 6-bromohexyl ester (1.40 g, 3.02 mmol) was dissolved in acetonitrile (10 mL), and ethanolamine (83.0 mg, 1.36 mmol), potassium carbonate (1.25 g, 9.06 mmol), potassium iodide (752 mg, 4.53 mmol), and tetrahydrofuran (4 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(Valeryloxy)dodecyl 6-[(2-hydroxyethyl)(6-oxyidene-6-{[2-(Valeryloxy)dodecyl]oxy}hexyl)amino]hexyl ester (670 mg, 25.2% yield) as a yellow liquid.

[1380] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.17-5.05 (m, 2H), 4.25 (dd, J = 3.2, 12.0Hz, 2H), 4.04 (dd, J = 6.8, 12.0Hz, 2H), 3.61 (br s, 2H), 2.67 (br s, 2H), 2.56 (br d,J=6.4Hz,4H),2.33(t,J=7.6Hz,8H),1.68-1.49(m,17H),1.44-1.26(m,40H),0.92(td,J=7.2,16.4Hz,12H).

[1381] LCMS:RT=2.013,m / z 826.5[M+H] + .

[1382] Preparation Example 41 Preparation of Compound 67

[1383] Step 1: Synthesis of 2-(benzyloxy)-3-(nonanoyloxy)propyl nonyl ester

[1384] Reaction formula:

[1385]

[1386] Material ratio:

[1387]

[1388] Operation process:

[1389] 2-(Benzyloxy)propane-1,3-diol (3.00 g, 16.5 mmol) was dissolved in dichloromethane (60 mL). Renyl acid (5.99 g, 37.9 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (7.89 g, 41.2 mmol), and 4-dimethylaminopyridine (402 mg, 3.29 mmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-(benzyloxy)-3-(nonanoyloxy)propyl nonanoate (6.00 g, 78.8% yield) as a colorless liquid.

[1390] Step 2: Synthesis of 2-hydroxy-3-(nonanoyloxy)propyl nonyl ester

[1391] Reaction formula:

[1392]

[1393] Material ratio:

[1394]

[1395] Operation process:

[1396] 2-(Benzyloxy)-3-(nonanoyloxy)propyl nonanate (6.00 g, 13.0 mmol) was dissolved in methanol (50 mL) and tetrahydrofuran (10 mL). Palladium / carbon (1.38 g, 1.30 mmol) was added under a nitrogen atmosphere. The mixture was reacted at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was filtered and concentrated. The crude product was purified by column chromatography to afford 2-hydroxy-3-(nonanoyloxy)propyl nonanate (3.80 g, 78.7% yield) as a colorless liquid.

[1397] Step 3: Synthesis of 1-[(6-bromohexanoyl)oxy]-3-(octanoyloxy)propan-2-ylnonyl ester

[1398] Reaction formula:

[1399]

[1400] Material ratio:

[1401]

[1402]

[1403] Operation process:

[1404] 2-Hydroxy-3-(nonanoyloxy)propyl nonanate (2.00 g, 5.37 mmol) was dissolved in dichloromethane (20 mL). 6-Bromohexanoic acid (1.36 g, 6.98 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.54 g, 8.05 mmol), and 4-dimethylaminopyridine (65.6 mg, 537 μmol) were added sequentially. The mixture was reacted at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-[(6-bromohexanoyl)oxy]-3-(octanoyloxy)propan-2-ylnonanate (2.50 g, 87.0% yield) as a colorless liquid.

[1405] Step 4: Synthesis of 1-{[6-({6-[2,3-bis(nonanoyloxy)propoxy]-6-oxyylidenehexyl}(2-hydroxyethyl)amino)hexanoyl]oxy}-3-(nonanoyloxy)propan-2-ylnonyl ester

[1406] Reaction formula:

[1407]

[1408] Material ratio:

[1409]

[1410] Operation process:

[1411] 1-[(6-bromohexanoyl)oxy]-3-(octanoyloxy)propan-2-ylnonanate (2.50 g, 4.67 mmol) was dissolved in acetonitrile (25 mL). Ethanolamine (175 mg, 2.33 mmol), potassium carbonate (2.10 g, 15.2 mmol), and potassium iodide (852 mg, 5.13 mmol) were added sequentially. The mixture was reacted at 70°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-{[6-({6-[2,3-bis(nonanoyloxy)propoxy]-6-oxyidenehexyl}(2-hydroxyethyl)amino)hexanoyl]oxy}-3-(nonanoyloxy)propan-2-ylnonanate (300 mg, 11.1% yield) as a pale yellow liquid.

[1412] 1 H NMR(400MHz, CHLOROFORM-d)δ=5.31-5.23(m,2H),4.32(dd,J=4.4,11.9Hz,4H),4.16(dd,J=5.9,11.9Hz,4H),3.54(t,J= 5.3Hz,2H),2.59(t,J=5.4Hz,2H),2.50-2.43(m,4H),2.37-2.29(m,12H),1.70-1.59(m,12H),1.51-1.43(m,4H),1.29(br d,J=5.3Hz,43H),1.02-0.77(m,12H)ppm.

[1413] LCMS:RT=2.135,m / z 824.6[M+H] + .

[1414] Preparation Example 42 Preparation of Compound 68

[1415] Step 1: Synthesis of 1-(benzyloxy)dodec-2-yl 6-bromohexyl ester

[1416] Reaction formula:

[1417]

[1418] Material ratio:

[1419]

[1420] Operation process:

[1421] 1-(Benzyloxy)dodecan-2-ol (2.50 g, 8.55 mmol) and 6-bromohexanoic acid (2.00 g, 10.2 mmol) were dissolved in dichloromethane (40 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.46 g, 12.8 mmol) and 4-dimethylaminopyridine (104 mg, 0.854 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodecan-2-yl 6-bromohexyl ester (3.18 g, 79.2% yield) as a colorless liquid.

[1422] Step 2: Synthesis of 1-hydroxydodecane-2-yl 6-bromohexyl ester

[1423] Reaction formula:

[1424]

[1425] Material ratio:

[1426]

[1427] Operation process:

[1428] Dissolve 1-(Benzyloxy)dodec-2-yl 6-bromohexyl ester (3.18 g, 6.77 mmol) in methanol (32 mL), add palladium on carbon (1.44 g, 1.35 mmol), and react under hydrogen at 35°C, 40 Psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain a colorless liquid, 1-hydroxydodec-2-yl 6-bromohexyl ester (1.88 g, 73.1% yield).

[1429] Step 3: Synthesis of 2-[(6-bromohexanoyl)oxy]dodecyl octyl ester

[1430] Reaction formula:

[1431]

[1432] Material ratio:

[1433]

[1434]

[1435] Operation process:

[1436] 1-Hydroxydodec-2-yl 6-bromohexyl ester (1.88 g, 4.96 mmol) and octanoic acid (857 mg, 5.95 mmol) were dissolved in dichloromethane (40 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.42 g, 7.43 mmol) and 4-dimethylaminopyridine (60.5 mg, 0.495 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to obtain 2-[(6-bromohexanoyl)oxy]dodecyloctyl ester (2.28 g, 91.0% yield) as a colorless liquid.

[1437] Step 4: Synthesis of 2-({6-[(2-hydroxyethyl)(6-{[1-(octanoyloxy)dodec-2-yl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodecyl octyl ester

[1438] Reaction formula:

[1439]

[1440] Material ratio:

[1441]

[1442] Operation process:

[1443] 2-[(6-bromohexanoyl)oxy]dodecyl octyl ester (2.00 g, 1.78 mmol) was dissolved in acetonitrile (15 mL), and 2-aminoethane-1-ol (108 mg, 3.96 mmol), potassium carbonate (1.64 g, 11.8 mmol), potassium iodide (985 mg, 5.93 mmol), and tetrahydrofuran (5 mL) were added in sequence. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-({6-[(2-hydroxyethyl)(6-{[1-(octanoyloxy)dodec-2-yl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodecyl octyl ester (840 mg, 20.6% yield) as a yellow liquid.

[1444] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J = 3.2, 6.4Hz, 2H), 4.25 (dd, J = 3.2, 12.0Hz, 2H), 4.05 (dd, J = 6.4, 12.0Hz, 2H), 3.64 (br s, 2H), 2.71 (br s,2H),2.60(brs,4H),2.33(dt,J=4.8,7.6Hz,8H),1.71-1.51(m,17H),1.40-1.24(m,52H),0.96-0.84(m,12H).

[1445] LCMS:RT=2.128,m / z 910.8[M+H] + .

[1446] Preparation Example 43 Preparation of Compound 62

[1447] Step 1: Preparation of 62-1

[1448] Reaction formula:

[1449]

[1450] Material ratio:

[1451] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Hydroxymethyl-1,3-propanediol 106 1eq 3g 30 Hexanoic acid 116 2 eq 6.5g 60 EDCI 192 2.5eq 14g 75 DMAP 122 0.25 eq 900mg 7.5 DCM - - 200mL -

[1452] Operation process:

[1453] 2-Hydroxymethyl-1,3-propanediol, hexanoic acid, EDCI, DMAP, and DCM were added to a reaction flask and allowed to react at room temperature for 2 h. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was washed once with 200 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 3.5 g of an oil.

[1454] Step 2: Preparation of 62-2

[1455] Reaction formula:

[1456]

[1457] Material ratio:

[1458] Material Name Molecular weight Feed ratio Feeding amount mmol 62-1 302 1.2eq 3.5g 11.6 7-Bromoheptanoic acid 209 1eq 2g 9.7 EDCI 192 1.5eq 3g 15 DMAP 122 0.5 eq 200mg 1.5 DCM - - 60mL -

[1459] Operation process:

[1460] To the reaction flask, 62-1, 7-bromoheptanoic acid, EDCI, DMAP, and DCM were added and allowed to react at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 60 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 4.2 g of an oil.

[1461] Step 3: Preparation of compound 62-3

[1462] Reaction formula:

[1463]

[1464] Material ratio:

[1465] Material Name Molecular weight Feed ratio Feeding amount mmol 5-Bromopentanol 167 1 eq 3.34g 20 p-Nitrophenyl chloroformate 201 1.2 eq 4.8g 24 Pyridine 79 1.5 eq 2.4g 30 DCM - - 60mL -

[1466] Operation process:

[1467] 5-Bromopentanol, p-nitrophenyl chloroformate, pyridine, and DCM were added to the reaction flask and allowed to react at room temperature for 16 h. TLC (PE:EA = 4:1) revealed an Rf value of 0.6 for the product. Purification by column chromatography yielded 5.3 g of a colorless oil.

[1468] Step 4: Preparation of compound 62-4

[1469] Reaction formula:

[1470]

[1471] Material ratio:

[1472] Material Name Molecular weight Feed ratio Feeding amount mmol 62-3 332 1 eq 3.32g 10 Undecanol 172 4eq 6.88g 40 Pyridine 79 1.2 eq 950mg 12 DMAP 122 0.2 eq 250mg 2 DCM - - 60mL -

[1473] Operation process:

[1474] To the reaction flask, 62-3, undecanol, pyridine, DMAP, and DCM were added and allowed to react at room temperature for 16 h. TLC (PE:EA = 10:1) revealed an Rf value of 0.6 for the product. Purification by column chromatography yielded 2.8 g of a colorless oil.

[1475] Step 5: Preparation of compound 62-5

[1476] Reaction formula:

[1477]

[1478] Material ratio:

[1479] Material Name Molecular weight Feed ratio Feeding amount mmol 62-4 365 1eq 2.8g 7.67 Ethanolamine 61 10 eq 4.7g 77 Acetonitrile - - 60mL -

[1480] Operation process:

[1481] Compound 62-4, ethanolamine, and acetonitrile were added to the reaction flask and reacted at room temperature for 16 h. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.3 for the product. Purification by column chromatography afforded 1.8 g of a colorless oil.

[1482] Step 6: Preparation of compound 62

[1483] Reaction formula:

[1484]

[1485] Material ratio:

[1486] Material Name Molecular weight Feed ratio Feeding amount mmol 62-5 345 1eq 1.8g 5.2 62-2 493 1.2eq 3g 6.3 potassium carbonate 138 1.5 eq 1.1g 7.8 Potassium iodide 166 0.5 eq 430mg 2.6 Acetonitrile - - 50mL -

[1487] Operation process:

[1488] Add 62-5, 62-2, potassium carbonate, potassium iodide, and acetonitrile to a reaction flask and react at 65°C for 16 h. TLC (DCM:MeOH = 10:1) reveals an Rf value of 0.5 for the product. Dilute the reaction solution with 100 mL of ethyl acetate and wash once with 100 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.5 g of an oil.

[1489] 1 H NMR(400MHz,Chloroform-d)δ4.12(dd,J=8.7,6.4Hz,10H),3.57(t,J=5.3Hz,2H),2.63(t,J=5.3Hz,2H),2.55–2.47(m,4H),2.39(p,J =6.0Hz,1H),2.31(td,J=7.6,1.9Hz,6H),1.72–1.58(m,10H),1.50(dp,J=14.6,7.3Hz,4H),1.38–1.23(m,30H),0.88(q,J=6.7Hz,9H).

[1490] MS (ES+) m / z): 758.6 (M+H) + .

[1491] Preparation Example 44 Preparation of Compound 63

[1492] Step 1: Synthesis of (4S)-4-[(1E)-dec-1-en-1-yl]-2,2-dimethyl-1,3-dioxolane

[1493] Reaction formula:

[1494]

[1495] Material ratio:

[1496]

[1497] Operation process:

[1498] Nonyltriphenylphosphane bromide (19.0 g, 40.4 mmol) was dissolved in tetrahydrofuran (190 mL). n-Butyllithium (2.5 M, 17.8 mL) was added at -20°C and reacted for half an hour. (4R)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde (5.54 g, 40.4 mmol) was added to the reaction solution at -20°C. The reaction was carried out at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction was quenched and extracted twice with ethyl acetate. The organic phase was concentrated and purified by column chromatography to give a colorless liquid (4S)-4-[(1E)-dec-1-en-1-yl]-2,2-dimethyl-1,3-dioxolane (6.37 g, 65.4% yield).

[1499] Step 2: Synthesis of (4S)-4-decyl-2,2-dimethyl-1,3-dioxolane

[1500] Reaction formula:

[1501]

[1502] Material ratio:

[1503]

[1504] Operation process:

[1505] Dissolve (4S)-4-decyl-2,2-dimethyl-1,3-dioxolane (6.37 g, 26.5 mmol) in methanol (60.0 mL), add palladium carbon (2.82 g, 2.65 mmol), and react under a hydrogen atmosphere at 25°C, 30 Psi for 12 hours. The reaction is complete after TLC monitoring. The mixture is filtered and concentrated to give a colorless liquid (4S)-4-decyl-2,2-dimethyl-1,3-dioxolane (6.42 g, 100% yield).

[1506] Step 3: Synthesis of (2R)-dodecane-1,2-diol

[1507] Reaction formula:

[1508]

[1509] Material ratio:

[1510]

[1511] Operation process:

[1512] (4S)-4-Decyl-2,2-dimethyl-1,3-dioxolane (6.42 g, 26.4 mmol) was dissolved in tetrahydrofuran (64.0 mL), and hydrochloric acid (3 M, 6.40 mL) was added. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After adjusting the pH to neutral, the mixture was extracted with ethyl acetate and the organic phase was concentrated. Purification by column chromatography afforded (2R)-dodecane-1,2-diol (4.43 g, 82.6% yield) as a white solid.

[1513] Step 4: Synthesis of (2R)-1-[(tert-butyldimethylsilyl)oxy]dodecan-2-ol

[1514] Reaction formula:

[1515]

[1516] Material ratio:

[1517] Material Name Molecular weight Feed ratio Feeding amount mmol (2R)-Dodecane-1,2-diol 202 1.00 eq 2.00g 9.88 tert-Butylchlorodimethylsilane 150 1.05eq 1.56g 10.3 Imidazole 68 1.05eq 0.706g 10.3 Tetrahydrofuran - - 20.0mL -

[1518] Operation process:

[1519] (2R)-Dodecane-1,2-diol (2.00 g, 9.88 mmol) and imidazole (706 mg, 10.3 mmol) were dissolved in tetrahydrofuran (20.0 mL). tert-Butylchlorodimethylsilane (1.56 g, 10.3 mmol) was slowly added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography afforded a colorless liquid, (2R)-1-[(tert-butyldimethylsilyl)oxy]dodecan-2-ol (2.40 g, 76.6% yield).

[1520] Step 5: Synthesis of (2R)-1-[(tert-butyldimethylsilyl)oxy]dodec-2-yloctyl ester

[1521] Reaction formula:

[1522]

[1523] Material ratio:

[1524]

[1525] Operation process:

[1526] (2R)-1-[(tert-Butyldimethylsilyl)oxy]dodecan-2-ol (2.40 g, 7.58 mmol) and n-octanoic acid (1.31 g, 9.10 mmol) were dissolved in dichloromethane (24.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.18 g, 11.3 mmol) and 4-dimethylaminopyridine (92.6 mg, 758 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave (2R)-1-[(tert-Butyldimethylsilyl)oxy]dodecan-2-yloctyl ester (3.00 g, 89.3% yield) as a colorless liquid.

[1527] Step 6: Synthesis of (2R)-1-hydroxydodec-2-yloctyl ester

[1528] Reaction formula:

[1529]

[1530] Material ratio:

[1531]

[1532] Operation process:

[1533] (2R)-1-[(tert-Butyldimethylsilyl)oxy]dodec-2-yloctyl ester (3.00 g, 6.78 mmol) was dissolved in tetrahydrofuran (30.0 mL). Triethylamine hydrofluoride (3.28 g, 20.3 mmol) was added at 0°C. The reaction was allowed to proceed under nitrogen at 25°C for 12 hours. TLC revealed the formation of new spots. After adjusting the pH to neutral, the reaction was extracted with ethyl acetate and the organic phase was concentrated. Purification by column chromatography afforded (2R)-1-hydroxydodec-2-yloctyl ester (1.82 g, 81.7% yield) as a colorless liquid.

[1534] Step 7: Synthesis of (2R)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester

[1535] Reaction formula:

[1536]

[1537] Material ratio:

[1538]

[1539] Operation process:

[1540] (2R)-1-Hydroxydodec-2-yloctyl ester (1.82 g, 5.54 mmol) and 6-bromohexanoic acid (1.30 g, 6.65 mmol) were dissolved in dichloromethane (20.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.59 g, 8.31 mmol) and 4-dimethylaminopyridine (67.6 mg, 553 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave (2R)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester (2.49 g, 88.9% yield) as a colorless liquid.

[1541] Step 8: Synthesis of (2R)-1-({6-[(2-hydroxyethyl)(6-{[(2R)-2-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester

[1542] Reaction formula:

[1543]

[1544] Material ratio:

[1545]

[1546] Operation process:

[1547] (2R)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (57.9 mg, 949 μmol), potassium carbonate (956 mg, 6.92 mmol), potassium iodide (394 mg, 2.37 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave (2R)-1-({6-[(2-hydroxyethyl)(6-{[(2R)-2-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester (523 mg, 27.4% yield) as a colorless liquid.

[1548] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.4, J2 = 6.6Hz, 2H), 4.23 (dd, J1 = 3.4, J2 = 11.8Hz, 2H), 4.03 (dd, J1 = 6.8, J2 = 11.8Hz, 2H), 3.56 (br t,J=5.2Hz,2H),2.61(br t,J=4.8Hz,2H),2.49(br t,J=7.2Hz,4H),2.31(dt,J1=2.2,J2=7.6Hz,8H),1.65-1.55(m,12H),1.51-1.43(m,4H),1.34-1.17(m,52H),0.98-0.78(m,12H)ppm.

[1549] LCMS:RT=2.180,m / z 910.8[M+H] + .

[1550] Preparation Example 45 Preparation of Compound 64

[1551] Step 1: Synthesis of (4S)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde

[1552] Reaction formula:

[1553]

[1554] Material ratio:

[1555]

[1556] Operation process:

[1557] [(4R)-2,2-Dimethyl-1,3-dioxolane-4-yl]methanol (5.00 g, 37.8 mmol) was dissolved in acetone (25.0 mL). Trichloroisocyanuric acid (3.52 g, 15.1 mmol) and sodium acetate (3.72 g, 45.4 mmol) were added at 25°C. 1-Hydroxy-2,2,6,6-tetramethylpiperidine (14.8 mg, 94.5 μmol) was dissolved in acetone (15.0 mL) and slowly added to the reaction solution. The mixture was reacted at 59°C under nitrogen protection for 0.5 h. The reaction was completed after TLC monitoring. The reaction was filtered and the organic phase was concentrated to obtain a colorless liquid (4S)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde (7.00 g, crude product).

[1558] Step 2: Synthesis of nonyltriphenylphosphane bromide cation

[1559] Reaction formula:

[1560]

[1561] Material ratio:

[1562] Material Name Molecular weight Feed ratio Feeding amount mmol 1-Bromononane 206 1.00 eq 5.54g 40.4 Triphenylphosphine 262 1.05eq 19.0g 40.4 Toluene - - 200mL -

[1563] Operation process:

[1564] 1-Bromononane (20.0 g, 96.5 mmol) was dissolved in toluene (200 mL), and triphenylphosphine (26.5 g, 101 mmol) was added. The mixture was reacted at 115°C for 48 hours. The reaction was monitored by TLC. The mixture was cooled and diluted with isopropyl ether. The mixture was filtered to obtain nonyltriphenylphosphane bromide (28.0 g, 61.7% yield) as a white solid.

[1565] Step 3: Synthesis of (4R)-4-[(1E)-dec-1-en-1-yl]-2,2-dimethyl-1,3-dioxolane

[1566] Reaction formula:

[1567]

[1568] Material ratio:

[1569]

[1570] Operation process:

[1571] Nonyltriphenylphosphane bromide (9.96 g, 21.2 mmol) was dissolved in tetrahydrofuran (96.0 mL). n-Butyllithium (2.5 M, 8.48 mL) was added at -20°C and reacted for half an hour at -20°C. (4S)-2,2-dimethyl-1,3-dioxolane-4-carbaldehyde (5.54 g, 40.4 mmol) was added to the reaction solution at -20°C. The reaction was carried out at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC. The reaction was quenched and extracted twice with ethyl acetate. The organic phase was concentrated and purified by column chromatography to give a colorless liquid (4R)-4-[(1E)-dec-1-en-1-yl]-2,2-dimethyl-1,3-dioxolane (2.24 g, 50.5% yield).

[1572] Step 4: Synthesis of (4R)-4-decyl-2,2-dimethyl-1,3-dioxolane

[1573] Reaction formula:

[1574]

[1575] Material ratio:

[1576]

[1577] Operation process:

[1578] Dissolve ((4R)-4-[(1E)-dec-1-en-1-yl]-2,2-dimethyl-1,3-dioxolane (2.24 g, 9.32 mmol) in methanol (24.0 mL), add palladium carbon (991 mg, 931 μmol), and react under a hydrogen atmosphere at 25°C, 30 Psi for 12 hours. The reaction is monitored by TLC. The mixture is filtered and concentrated to give a colorless liquid (4R)-4-decyl-2,2-dimethyl-1,3-dioxolane (2.26 g, 100% yield).

[1579] Step 5: Synthesis of (2S)-dodecane-1,2-diol

[1580] Reaction formula:

[1581]

[1582] Material ratio:

[1583]

[1584] Operation process:

[1585] (4R)-4-Decyl-2,2-dimethyl-1,3-dioxolane (2.26 g, 9.32 mmol) was dissolved in tetrahydrofuran (30.0 mL), and hydrochloric acid (3 M, 2.25 mL) was added. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC analysis revealed the formation of new spots. After adjusting the pH to neutral, the mixture was extracted with ethyl acetate and the organic phase was concentrated. Purification by column chromatography afforded (2S)-dodecane-1,2-diol (1.49 g, 78.9% yield) as a white solid.

[1586] Step 6: Synthesis of (2S)-1-[(tert-butyldimethylsilyl)oxy]dodecan-2-ol

[1587] Reaction formula:

[1588]

[1589] Material ratio:

[1590]

[1591]

[1592] Operation process:

[1593] (2S)-Dodecane-1,2-diol (1.49 g, 7.36 mmol) and imidazole (526 mg, 7.73 mmol) were dissolved in tetrahydrofuran (20.0 mL). tert-Butylchlorodimethylsilane (1.17 g, 7.73 mmol) was slowly added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography afforded (2S)-1-[(tert-butyldimethylsilyl)oxy]dodecan-2-ol (2.00 g, 85.7% yield), a colorless liquid.

[1594] Step 7: Synthesis of (2S)-1-[(tert-butyldimethylsilyl)oxy]dodec-2-yloctyl ester

[1595] Reaction formula:

[1596]

[1597] Material ratio:

[1598]

[1599] Operation process:

[1600] (2S)-1-[(tert-Butyldimethylsilyl)oxy]dodecan-2-ol (2.00 g, 6.32 mmol) and n-octanoic acid (1.28 g, 8.84 mmol) were dissolved in dichloromethane (20.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.18 g, 11.3 mmol) and 4-dimethylaminopyridine (154 mg, 1.26 mmol) were added sequentially. The reaction was incubated at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave (2S)-1-[(tert-Butyldimethylsilyl)oxy]dodecan-2-yloctyl ester (2.50 g, 89.3% yield) as a colorless liquid.

[1601] Step 8: Synthesis of (2S)-1-hydroxydodec-2-yloctyl ester

[1602] Reaction formula:

[1603]

[1604] Material ratio:

[1605]

[1606] Operation process:

[1607] (2S)-1-[(tert-Butyldimethylsilyl)oxy]dodec-2-yloctyl ester (2.50 g, 5.65 mmol) was dissolved in tetrahydrofuran (25.0 mL). Triethylamine hydrofluoride (2.73 g, 16.9 mmol) was added at 0°C. The reaction was allowed to react at 25°C under nitrogen for 12 hours. TLC revealed the formation of new spots. After adjusting the pH to neutral, the reaction was extracted with ethyl acetate and the organic phase was concentrated. Purification by column chromatography afforded (2S)-1-hydroxydodec-2-yloctyl ester (1.37 g, 73.8% yield) as a colorless liquid.

[1608] Step 9: Synthesis of (2S)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester

[1609] Reaction formula:

[1610]

[1611] Material ratio:

[1612]

[1613] Operation process:

[1614] (2S)-1-Hydroxydodec-2-yloctyl ester (1.37 g, 4.17 mmol) and 6-bromohexanoic acid (1.14 g, 5.84 mmol) were dissolved in dichloromethane (13.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.44 g, 7.51 mmol) and 4-dimethylaminopyridine (101 mg, 834 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring revealed the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave (2S)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester (1.94 g, 92.0% yield) as a colorless liquid.

[1615] Step 10: Synthesis of (2S)-1-({6-[(2-hydroxyethyl)(6-{[(2S)-2-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester

[1616] Reaction formula:

[1617]

[1618] Material ratio:

[1619]

[1620]

[1621] Operation process:

[1622] (2S)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (10.0 mL). Ethanolamine (57.9 mg, 949 μmol), potassium carbonate (956 mg, 6.92 mmol), potassium iodide (394 mg, 2.37 mmol), and tetrahydrofuran (5.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave (2S)-1-({6-[(2-hydroxyethyl)(6-{[(2S)-2-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester (430 mg, 22.3% yield) as a colorless liquid.

[1623] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (br dd, J1 = 3.2, J2 = 6.2Hz, 2H), 4.23 (dd, J1 = 3.2, J2 = 11.8Hz, 2H), 4.03 (dd, J1 = 6.8, J2 = 11.8Hz, 2H), 3.56 (br t,J=5.2Hz,2H),2.61(br t,J=4.8Hz,2H),2.49(br t,J=7.2Hz,4H),2.31(dt,J1=2.2,J2=7.4Hz,8H),1.70-1.57(m,12H),1.51-1.43(m,4H),1.35-1.22(m,52H),0.89(t,J=6.6Hz,12H)ppm.

[1624] LCMS:RT=2.204,m / z 910.8[M+H] + .

[1625] Preparation Example 46 Preparation of Compound 65

[1626] Step 1: Synthesis of (2R)-1-({6-[(2-hydroxyethyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester

[1627] Reaction formula:

[1628]

[1629] Material ratio:

[1630]

[1631] Operation process:

[1632] Dissolve (2R)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester (1.15 g, 2.27 mmol) in acetonitrile (11.0 mL), followed by the addition of ethanolamine (1.39 g, 22.7 mmol). The mixture was reacted at 30°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, followed by filtration and concentration. Purification by column chromatography afforded a colorless liquid, (2R)-1-({6-[(2-hydroxyethyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester (740 mg, 66.9% yield).

[1633] Step 2: Synthesis of 2-(octanoyloxy)nonyl 8-[(2-hydroxyethyl)[6-(nonyloxy)-6-oxyylidenehexyl]amino]octyl ester

[1634] Reaction formula:

[1635]

[1636] Material ratio:

[1637]

[1638] Operation process:

[1639] (2R)-1-({6-[(2-hydroxyethyl)amino]hexanoyl}oxy)dodec-2-yloctyl ester (400 mg, 823 μmol) was dissolved in acetonitrile (4.00 mL). (2S)-1-[(6-bromohexanoyl)oxy]dodec-2-yloctyl ester (416 mg, 823 μmol), potassium carbonate (398 mg, 2.88 mmol), potassium iodide (164 mg, 988 μmol), and tetrahydrofuran (2.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(octanoyloxy)nonyl 8-[(2-hydroxyethyl)[6-(nonyloxy)-6-oxyylidenehexyl]amino]octyl ester (780 mg, 69.2% yield) as a colorless liquid.

[1640] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.18-5.00 (m, 2H), 4.23 (dd, J1 = 3.4, J2 = 11.8Hz, 2H), 4.03 (dd, J1 = 6.8, J2 = 11.8Hz, 2H), 3.59 (br t, J = 4.8Hz, 2H), 2.64 (br s,2H),2.52(br t,J=6.8Hz,4H),2.31(dt,J1=2.8,J2=7.6Hz,8H),1.64-1.44(m,16H),1.38-1.20(m,52H),0.89(t,J=6.8Hz,12H)ppm.LCMS:RT=2.212,m / z 910.7[M+H] + .

[1641] Preparation Example 47 Preparation of Compound 10

[1642] Step 1: Preparation of 10-1

[1643] Reaction formula:

[1644]

[1645] Material ratio:

[1646] Material Name Molecular weight Feed ratio Feeding amount mmol 4-Bromobutyric acid 167 1 eq 6.68g 40 1,2-Epoxydodecane 184 1eq 7.36g 40 Ferric chloride 162 0.05 eq 324mg 2 Pyridine 79 0.05 eq 158mg 2

[1647] Operation process:

[1648] 4-Bromobutyric acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to the reaction flask and allowed to react at room temperature for 16 hours. The product had an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction solution was concentrated and purified by column chromatography to yield 8 g of an oil.

[1649] Step 2: Preparation of 10-2

[1650] Reaction formula:

[1651]

[1652] Material ratio:

[1653] Material Name Molecular weight Feed ratio Feeding amount mmol 10-1 351.33 1eq 8g 22.8 bitter 144.21 1.1 eq 3.6g 25 EDCI 155 1.5eq 5.3g 34 DMAP 122 0.15 eq 420mg 3.4 DCM - - 150mL -

[1654] Operation process:

[1655] 10-1, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and reacted at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was concentrated and purified by column chromatography to obtain 7.5 g of an oil.

[1656] Step 3: Preparation of compound 10

[1657] Reaction formula:

[1658]

[1659] Material ratio:

[1660] Material Name Molecular weight Feed ratio Feeding amount mmol 10-2 477.5 2.2 eq 3.15g 6.6 Ethanolamine 61 1 eq 183mg 3 potassium carbonate 138 3eq 1.24g 9 Potassium iodide 166 1 eq 500mg 3 Acetonitrile - - 50mL -

[1661] Operation process:

[1662] 10-2, ethanolamine, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1 g of a colorless oil.

[1663] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.9,3.3Hz,2H),4.23(dd,J=11.8,3.3Hz,2H),4.01(dd,J=11.8,6.9Hz,2H),3.54(t,J=5.2Hz,2H),2.60(d,J =6.5Hz,2H),2.49(t,J=7.2Hz,4H),2.31(q,J=7.2Hz,8H),1.76(p,J=7.3Hz ,4H),1.65–1.52(m,12H),1.27(d,J=13.5Hz,44H),0.88(t,J=6.7Hz,12H).

[1664] MS (ES+) m / z): 854.7 (M) + .

[1665] Preparation Example 48 Preparation of Compound 12

[1666] Step 1: Synthesis of 1-(Benzyloxy)dodec-2-yl 7-methyloctyl ester

[1667] Reaction formula:

[1668]

[1669] Material ratio:

[1670]

[1671] Operation process:

[1672] Dissolve 1-(Benzyloxy)dodecan-2-ol (5.60 g, 19.1 mmol) and 7-methyloctanoic acid (3.03 g, 19.1 mmol) in dichloromethane (50.0 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.51 g, 28.7 mmol) and 4-dimethylaminopyridine (245 mg, 2.01 mmol) sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. Column chromatography of the crude product afforded 1-(Benzyloxy)dodecan-2-yl 7-methyloctyl ester (5.75 g, 64.4% yield) as a colorless liquid.

[1673] Step 2: Synthesis of 1-hydroxydodec-2-yl 7-methyloctyl ester

[1674] Reaction formula:

[1675]

[1676] Material ratio:

[1677] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodec-2-yl 7-methyloctyl ester 432 1.0 eq 2.87g 6.63 Palladium / Carbon 106 0.10eq 0.233g 1.91 hydrogen - (40Psi) - - Methanol - 50.0mL -

[1678] Operation process:

[1679] In two parallel batches, 1-(benzyloxy)dodec-2-yl 7-methyloctyl ester (2.87 g, 6.63 mmol) was dissolved in methanol (50.0 mL). Palladium / carbon (233 mg, 1.91 mmol) was added under an argon atmosphere. The mixture was reacted at 25°C under a hydrogen atmosphere (40 psi) for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. The mixture was spin-dried to give 1-hydroxydodec-2-yl 7-methyloctyl ester (3.65 g, 80.3% yield) as a colorless liquid.

[1680] Step 3: Synthesis of 1-[(6-bromohexanoyl)oxy]dodec-2-yl 7-methyloctyl ester

[1681] Reaction formula:

[1682]

[1683] Material ratio:

[1684]

[1685] Operation process:

[1686] Dissolve 1-hydroxydodec-2-yl 7-methyloctyl ester (1.20 g, 3.50 mmol) in dichloromethane (12.0 mL). 6-Bromohexanoic acid (956 mg, 4.90 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.01 g, 5.25 mmol), and 4-dimethylaminopyridine (42.8 mg, 350 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to afford 1-[(6-bromohexanoyl)oxy]dodec-2-yl 7-methyloctyl ester (1.57 g, 86.2% yield) as a colorless liquid.

[1687] Step 4: Synthesis of 1-({6-[(2-hydroxyethyl)[6-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl]amino]hexanoyl}oxy)dodec-2-yl 7-methyloctyl ester

[1688] Reaction formula:

[1689]

[1690] Material ratio:

[1691]

[1692] Operation process:

[1693] 1-[(6-bromohexanoyl)oxy]dodec-2-yl 7-methyloctyl ester (200 mg, 384 μmol) was dissolved in acetonitrile (2.00 mL). Ethanolamine (11.2 mg, 184 μmol), potassium carbonate (186 mg, 1.35 mmol), potassium iodide (76.6 mg, 461 μmol), and tetrahydrofuran (1.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({6-[(2-hydroxyethyl)[6-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl]amino]hexanoyl}oxy)dodec-2-yl 7-methyloctyl ester (98.0 mg, 27.1% yield) as a colorless liquid.

[1694] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.17-4.91 (m, 2H), 4.24 (dd, J1 = 3.0, J2 = 11.6Hz, 2H), 4.02 (ddd, J1 = 3.2, J2 = 6.8, J3 = 11.8Hz, 2H), 3.63 (br s,2H),2.73-2.49(m,5H),2.40-2.25(m,6H),2.19-1.97(m,4H),1.63(br dd, J1=7.6, J2=15.2Hz,12H),1.41-1.09(m,42H),1.01-0.75(m,30H)ppm.

[1695] LCMS:RT=2.182,m / z 960.8[M+23] + .

[1696] Preparation Example 49 Preparation of Compound 13

[1697] Step 1: Synthesis of 1-[(4-bromobutyryl)oxy]dodec-2-yl 7-methyloctyl ester

[1698] Reaction formula:

[1699]

[1700] Material ratio:

[1701]

[1702] Operation process:

[1703] Dissolve 1-hydroxydodec-2-yl 7-methyloctyl ester (1.20 g, 3.50 mmol) in dichloromethane (12.0 mL). 4-Bromobutyric acid (819 mg, 4.90 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.01 g, 5.25 mmol), and 4-dimethylaminopyridine (42.8 mg, 350 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to afford 1-[(4-bromobutyryl)oxy]dodec-2-yl 7-methyloctyl ester (1.20 g, 69.6% yield) as a colorless liquid.

[1704] Step 2: Synthesis of 1-({4-[(2-hydroxyethyl)[4-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-4-oxyylidenebutyl]amino]butyryl}oxy)dodec-2-yl 7-methyloctyl ester

[1705] Reaction formula:

[1706]

[1707] Material ratio:

[1708]

[1709]

[1710] Operation process:

[1711] 1-[(4-bromobutyryl)oxy]dodec-2-yl 7-methyloctyl ester (200 mg, 406 μmol) was dissolved in acetonitrile (2.00 mL). Ethanolamine (11.9 mg, 195 μmol), potassium carbonate (196 mg, 1.42 mmol), potassium iodide (81.0 mg, 488 μmol), and tetrahydrofuran (1.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({4-[(2-hydroxyethyl)[4-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-4-oxyylidenebutyl]amino]butyryl}oxy)dodec-2-yl 7-methyloctyl ester (84.0 mg, 23.4% yield) as a colorless liquid.

[1712] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.23-4.92 (m, 2H), 4.25 (dd, J1 = 3.0, J2 = 11.8Hz, 2H), 4.09-3.92 (m, 2H), 3.56 (br s,2H),2.71-2.43(m,6H),2.40-2.25(m,6H),2.20-1.99(m,4H),1.88-1. 70(m,4H),1.68-1.60(m,2H),1.42-1.07(m,38H),1.04-0.79(m,30H)ppm.

[1713] LCMS:RT=2.143,m / z 882.6[M+H] + .

[1714] Preparation Example 50 Preparation of Compound 14

[1715] Step 1: Synthesis of 1-[(8-bromooctanoyl)oxy]dodec-2-yl 7-methyloctyl ester

[1716] Reaction formula:

[1717]

[1718] Material ratio:

[1719]

[1720] Operation process:

[1721] Dissolve 1-hydroxydodec-2-yl 7-methyloctyl ester (1.20 g, 3.50 mmol) in dichloromethane (12.0 mL). 8-bromooctanoic acid (1.09 g, 4.90 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.01 g, 5.25 mmol), and 4-dimethylaminopyridine (42.8 mg, 350 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. Column chromatography of the crude product afforded 1-[(8-bromooctanoyl)oxy]dodec-2-yl 7-methyloctyl ester (1.87 g, 97.4% yield) as a colorless liquid.

[1722] Step 2: Synthesis of 1-({8-[(2-hydroxyethyl)[8-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-8-oxyylideneoctyl]amino]octanoyl}oxy)dodec-2-yl 7-methyloctyl ester

[1723] Reaction formula:

[1724]

[1725] Material ratio:

[1726]

[1727] Operation process:

[1728] 1-[(8-Bromooctanoyl)oxy]dodec-2-yl 7-methyloctyl ester (200 mg, 365 μmol) was dissolved in acetonitrile (2.00 mL). Ethanolamine (10.7 mg, 175 μmol), potassium carbonate (176 mg, 1.28 mmol), potassium iodide (72.7 mg, 438 μmol), and tetrahydrofuran (1.00 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({8-[(2-hydroxyethyl)[8-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-8-oxyylideneoctyl]amino]octanoyl}oxy)dodec-2-yl 7-methyloctyl ester (112 mg, 30.8% yield) as a colorless liquid.

[1729] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.21-4.94 (m, 2H), 4.35-4.14 (m, 2H), 4.03 (ddd, J1 = 2.8, J2 = 6.8, J3 = 11.8Hz, 2H), 3.69 (br s,2H),2.79-2.51(m,5H),2.37-2.25(m,6H),2.19-1.97(m,5H),1.66-1.57(m,9H),1.39-1.05(m,51H),1.03-0.80(m,30H)ppm.

[1730] LCMS:RT=2.209,m / z 995.2[M+H] + .

[1731] Preparation Example 51 Preparation of Compound 15

[1732] Step 1: Synthesis of heptadecan-9-yl 8-[(2-hydroxyethyl)(8-{[2-(octanoyloxy)undecyl]oxy}-8-oxyylideneoctyl)amino]octyl ester

[1733] Reaction formula:

[1734]

[1735] Material ratio:

[1736]

[1737]

[1738] Operation process:

[1739] 2-(Octanoyloxy)undecyl 8-bromooctyl ester (50.0 mg, 0.096 mmol) was dissolved in acetonitrile (0.3 mL). Heptadec-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (35.4 mg, 0.08 mmol), potassium carbonate (38.7 mg, 0.28 mmol), potassium iodide (15.9 mg, 0.096 mmol), and tetrahydrofuran (0.2 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave heptadec-9-yl 8-[(2-hydroxyethyl)(8-{[2-(octanoyloxy)undecyl]oxy}-8-oxyylideneoctyl)amino]octyl ester (33 mg, 46.2% yield).

[1740] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.08 (dq, J1 = 3.2, J2 = 6.8Hz, 1H), 4.87 (quin, J = 6.0H z,1H),4.22(dd,J1=3.2,J2=12.0Hz,1H),4.04(dd,J1=6.8,J2=12.0Hz,1H),3.58(br t,J=5.2Hz,2H),2.65(br t,J=4.4Hz,2H),2.55-2.48(m,3H),2.30(q,J=8.0Hz,6H),1.63(br d,J=6.8Hz,10H),1.53-1.42(m,9H),1.37-1.19(m,57H),0.89(t,J=6.8Hz,12H).

[1741] LCMS:RT=2.222,m / z 880.7[M+H] + .

[1742] Preparation Example 52 Preparation of Compound 16

[1743] Step 1: Synthesis of 1-(Benzyloxy)undec-2-yloctyl ester

[1744] Reaction formula:

[1745]

[1746] Material ratio:

[1747]

[1748] Operation process:

[1749] 1-(Benzyloxy)undecan-2-ol (2.0 g, 7.18 mmol) and octanoic acid (1.24 g, 8.62 mmol) were dissolved in dichloromethane (20 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.07 g, 10.7 mmol) and 4-dimethylaminopyridine (87.7 mg, 0.718 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)undecan-2-yloctyl ester (1.56 g, 53.6% yield) as a colorless liquid.

[1750] Step 2: Synthesis of 1-hydroxyundecane-2-yloctyl ester

[1751] Reaction formula:

[1752]

[1753] Material ratio:

[1754] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)undec-2-yloctyl ester 404 1.0 eq 1.56g 3.86 Palladium on carbon 105.9 0.2 eq 820mg 0.771 Methanol - - 20mL -

[1755] Operation process:

[1756] Dissolve 1-(Benzyloxy)undec-2-yloctyl ester (1.56 g, 3.86 mmol) in methanol (20 mL), add palladium on carbon (820 mg, 0.771 mmol), and react under hydrogen at 35°C, 40 psi, for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1-hydroxyundec-2-yloctyl ester (600 mg, 49.4% yield), a colorless liquid.

[1757] Step 3: Synthesis of 2-(octanoyloxy)undecyl 8-bromooctyl ester

[1758] Reaction formula:

[1759]

[1760] Material ratio:

[1761]

[1762] Operation process:

[1763] 1-Hydroxyundec-2-yloctyl ester (600 mg, 1.91 mmol) and 8-bromooctanoic acid (510 mg, 2.29 mmol) were dissolved in dichloromethane (10 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (548 mg, 2.86 mmol) and 4-dimethylaminopyridine (23.3 mg, 0.19 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-(octanoyloxy)undecyl 8-bromooctyl ester (846 mg, 85.3% yield) as a colorless liquid.

[1764] Step 4: Synthesis of 2-(4-pentylcyclohexyl)ethyl 8-bromooctyl ester

[1765] Reaction formula:

[1766]

[1767] Material ratio:

[1768]

[1769]

[1770] Operation process:

[1771] 2-(4-Pentylcyclohexyl)ethan-1-ol (1.0 g, 5.04 mmol) and 8-bromooctanoic acid (1.35 g, 6.05 mmol) were dissolved in dichloromethane (15 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.45 g, 1.45 g) and 4-dimethylaminopyridine (61.5 mg, 0.504 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to yield 2-(4-pentylcyclohexyl)ethyl 8-bromooctyl ester (2.0 g, 98.3% yield), a colorless liquid.

[1772] Step 5: Synthesis of 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)amino]octyl ester

[1773] Reaction formula:

[1774]

[1775] Material ratio:

[1776] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(4-pentylcyclohexyl)ethyl 8-bromooctyl ester 402 1.0 eq 2.00g 4.96 Ethanolamine 61 10.0eq 3.03g 49.5 Acetonitrile - - 20mL -

[1777] Operation process:

[1778] Undecyl 8-bromooctyl ester (2.00 g, 4.96 mmol) was dissolved in acetonitrile (20 mL). Ethanolamine (3.03 g, 49.5 mmol) was added to the reaction mixture. The mixture was allowed to react at 25°C under nitrogen for 24 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Column chromatography of the crude product afforded 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)amino]octyl ester (1.23 g, 64.0% yield) as a bright yellow liquid.

[1779] Step 6: Synthesis of 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)(8-{[2-(octanoyloxy)undecyl]oxy}-8-oxyylideneoctyl)amino]octyl ester

[1780] Reaction formula:

[1781]

[1782] Material ratio:

[1783]

[1784]

[1785] Operation process:

[1786] 2-(Octanoyloxy)undecyl 8-bromooctyl ester (50.0 mg, 0.096 mmol) was dissolved in acetonitrile (0.3 mL). 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)amino]octyl ester (30.7 mg, 0.08 mmol), potassium carbonate (38.7 mg, 0.28 mmol), potassium iodide (15.9 mg, 0.096 mmol), and tetrahydrofuran (0.2 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a colorless liquid, 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)(8-{[2-(octanoyloxy)undecyl]oxy}-8-oxyylideneoctyl)amino]octyl ester (33 mg, 50.0% yield).

[1787] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.10 (dq, J1 = 4.0, J2 = 6.8Hz, 1H), 4.24 (dd, J1 = 3. 6,J2=12.0Hz,1H),4.15-4.09(m,2H),4.05(dd,J1=6.8,J2=12.0Hz,1H),3.60(br s,2H),2.73-2.60(m,2H),2.54(br s,3H),2.36-2.28(m,6H),1.76(br d,J=10.8Hz,5H),1.62(br d,J=6.0Hz,17H),1.38-1.23(m,44H),1.20-1.15(m,2H),0.90(br t, J = 6.8 Hz, 9H) ppm.

[1788] LCMS:RT=2.103,m / z 822.7[M+H] + .

[1789] Preparation Example 53 Preparation of Compound 17

[1790] Step 1: Preparation of 17-1

[1791] Reaction formula:

[1792]

[1793] Material ratio:

[1794] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mL 2.5

[1795] Operation process:

[1796] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[1797] Step 2: Preparation of 17-2

[1798] Reaction formula:

[1799]

[1800] Material ratio:

[1801] Material Name Molecular weight Feed ratio Feeding amount mmol 17-1 379.38 1 eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[1802] Operation process:

[1803] 17-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated aqueous sodium bicarbonate solution, and once with 100 mL of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[1804] Step 3: Preparation of 17-3

[1805] Reaction formula:

[1806]

[1807] Material ratio:

[1808] Material Name Molecular weight Feed ratio Feeding amount mmol 17-2 505.58 1eq 4g 7.9 Ethanolamine 61 10 eq 4.8g 79 Acetonitrile - - 60mL -

[1809] Operation process:

[1810] 17-2, ethanolamine, and acetonitrile were added to the reaction flask and reacted at room temperature for 16 hours. The product had an rR value of 0.3 according to TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.8 g of an oil.

[1811] Step 4: Preparation of 17-4

[1812] Reaction formula:

[1813]

[1814] Material ratio:

[1815] Material Name Molecular weight Feed ratio Feeding amount mmol 4-Bromobutyric acid 167 1 eq 6.68g 40 1,2-Epoxydodecane 184 1eq 7.36g 40 Ferric chloride 162 0.05 eq 324mg 2 Pyridine 79 0.05 eq 158mg 2

[1816] Operation process:

[1817] 4-Bromobutyric acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to the reaction flask and allowed to react at room temperature for 16 hours. The product had an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction solution was concentrated and purified by column chromatography to yield 8 g of an oil.

[1818] Step 5: Preparation of 17-5

[1819] Reaction formula:

[1820]

[1821] Material ratio:

[1822] Material Name Molecular weight Feed ratio Feeding amount mmol 17-4 351.33 1eq 8g 22.8 bitter 144.21 1.1 eq 3.6g 25 EDCI 155 1.5eq 5.3g 34 DMAP 122 0.15 eq 420mg 3.4 DCM - - 150mL -

[1823] Operation process:

[1824] 17-4, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and reacted at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was concentrated and purified by column chromatography to obtain 7.5 g of an oil.

[1825] Step 6: Preparation of compound 17

[1826] Reaction formula:

[1827]

[1828] Material ratio:

[1829] Material Name Molecular weight Feed ratio Feeding amount mmol 17-3 485.75 1eq 2.4g 5 17-5 477.52 1.2 eq 2.9g 6 potassium carbonate 138 1.5eq 1g 7.5 Potassium iodide 166 0.5 eq 420mg 2.5 Acetonitrile - - 50mL -

[1830] Operation process:

[1831] To a reaction flask, 17-3, 17-5, K2CO3, KI, and acetonitrile were added and heated to 65°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2 g of a colorless oil.

[1832] 1H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.2Hz,2H),4.22(ddd,J=11.9,5.6,3 .4Hz,2H),4.02(ddd,J=11.8,6.8,2.1Hz,2H),3.53(t,J=5.3Hz,2H),2.58(t,J=5.4H z,2H),2.47(dt,J=14.4,7.2Hz,4H),2.31(q,J=7.1Hz,8H),1.76(p,J=7.4Hz,2H),1. 65–1.53(m,12H),1.44(q,J=7.6Hz,2H),1.32–1.23(m,48H),0.88(t,J=6.7Hz,12H).

[1833] MS (ES+) m / z): 882.7 (M) + .

[1834] Preparation Example 54 Preparation of Compound 61

[1835] Step 1: Preparation of 61-1

[1836] Reaction formula:

[1837]

[1838] Material ratio:

[1839] Material Name Molecular weight Feed ratio Feeding amount mmol 5-Bromopentanol 167 1 eq 3.34g 20 p-Nitrophenyl chloroformate 201 1.2 eq 4.8g 24 Pyridine 79 1.5 eq 2.4g 30 DCM - - 60mL -

[1840] Operation process:

[1841] 5-Bromopentanol, p-nitrophenyl chloroformate, pyridine, and DCM were added to the reaction flask and allowed to react at room temperature for 16 h. TLC (PE:EA = 4:1) revealed an Rf value of 0.6 for the product. Purification by column chromatography yielded 5.3 g of a colorless oil.

[1842] Step 2: Preparation of 61-2

[1843] Reaction formula:

[1844]

[1845] Material ratio:

[1846] Material Name Molecular weight Feed ratio Feeding amount mmol 61-1 332 1 eq 3.32g 10 Undecanol 172 4eq 6.88g 40 Pyridine 79 1.2 eq 950mg 12 DMAP 122 0.2 eq 250mg 2 DCM - - 60mL -

[1847] Operation process:

[1848] To the reaction flask, 61-1, undecanol, pyridine, DMAP, and DCM were added and allowed to react at room temperature for 16 h. TLC (PE:EA = 10:1) revealed an Rf value of 0.6 for the product. Purification by column chromatography yielded 2.8 g of a colorless oil.

[1849] Step 3: Preparation of 61-3

[1850] Reaction formula:

[1851]

[1852] Material ratio:

[1853]

[1854]

[1855] Operation process:

[1856] 61-2, ethanolamine, and acetonitrile were added to the reaction flask and reacted at room temperature for 16 h. TLC (DCM:MeOH = 10:1) showed an Rf value of 0.3 for the product. Purification by column chromatography gave 1.8 g of a colorless oil.

[1857] Step 4: Preparation of 61-4

[1858] Reaction formula:

[1859]

[1860] Material ratio:

[1861] Material Name Molecular weight Feed ratio Feeding amount mmol glycerin 92 1eq 9.2g 100 Imidazole 68 2.5eq 17g 250 tert-Butyldimethylsilyl chloride 150.72 2.1eq 31.7g 210 Tetrahydrofuran - - 200mL -

[1862] Operation process:

[1863] Glycerol, imidazole, and tetrahydrofuran were added to the reaction flask, cooled to 0-5°C, and a solution of tert-butyldimethylsilyl chloride in tetrahydrofuran was added dropwise. The reaction was stirred at room temperature for 16 hours. TLC (PE:EA = 10:1) revealed an Rf value of 0.3 for the product. 500 mL of water was added to the reaction solution, followed by extraction with 500 mL of ethyl acetate. The organic phase was dried, concentrated, and purified by column chromatography to yield 28 g of a colorless oil.

[1864] Step 5: Preparation of 61-5

[1865] Reaction formula:

[1866]

[1867] Material ratio:

[1868] Material Name Molecular weight Feed ratio Feeding amount mmol 61-4 320.62 1 eq 12.8g 40 8-Bromooctanoic acid 223.11 1.1 eq 9.8g 44 EDCI 192 2eq 15.4g 80 DMAP 122 0.2eq 1g 8 dichloromethane - - 200mL -

[1869] Operation process:

[1870] 61-4,8-Bromooctanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and allowed to react at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 200 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 13.2 g of a colorless oil.

[1871] Step 6: Preparation of 61-6

[1872] Reaction formula:

[1873]

[1874] Material ratio:

[1875] Material Name Molecular weight Feed ratio Feeding amount mmol 61-5 525.72 1 eq 13.2g 25 1M tetrabutylammonium fluoride solution in tetrahydrofuran - 3eq 75mL 75 Tetrahydrofuran - - 100mL -

[1876] Operation process:

[1877] To the reaction flask, 61-5 and tetrahydrofuran were added, followed by a 1 M solution of tetrabutylammonium fluoride in tetrahydrofuran. The reaction was stirred at room temperature for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction solution was extracted with 400 mL of water and 400 mL of ethyl acetate. The organic phase was dried, concentrated, and purified by column chromatography to yield 6.5 g of a colorless oil.

[1878] Step 7: Preparation of 61-7

[1879] Reaction formula:

[1880]

[1881] Material ratio:

[1882] Material Name Molecular weight Feed ratio Feeding amount mmol 61-6 297.19 1eq 6g 20 Hexanoic acid 116 2.2 eq 5.1g 44 EDCI 192 3eq 11.5g 60 DMAP 122 0.2 eq 488mg 4 DCM - - 100mL -

[1883] Operation process:

[1884] To the reaction flask, 61-6, hexanoic acid, EDCI, DMAP, and DCM were added and stirred at room temperature for 16 hours. TLC (PE:EA = 20:1) revealed an Rf value of 0.6 for the product. The reaction mixture was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 8.2 g of a colorless oil.

[1885] Step 8: Preparation of compound 61

[1886] Reaction formula:

[1887]

[1888] Material ratio:

[1889]

[1890]

[1891] Operation process:

[1892] To a reaction flask, add 61-3, 61-7, potassium carbonate, potassium iodide, and acetonitrile and react at 65°C for 16 h. TLC (DCM:MeOH = 10:1) reveals an Rf value of 0.5 for the product. Dilute the reaction solution with 100 mL of ethyl acetate and wash once with 100 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.5 g of an oil.

[1893] 1 H NMR(400MHz,Chloroform-d)δ5.29–5.23(m,1H),4.29(dd,J=11.9,4.3Hz,2H),4.17–4.08(m,6H),3.61(t,J=5.3Hz,2H),2.68(t, J=5.4Hz,2H),2.56(q,J=7.2Hz,4H),2.31(td,J=7.6,2.6Hz,6H),1.74–1.44(m,14H),1.40–1.23(m,32H),0.88(q,J=6.8Hz,9H).

[1894] MS (ES+) m / z): 758.6 (M+H) + .

[1895] Example 1 Preparation and Detection of Lipid Nanoparticles (LNP)

[1896] To verify whether the lipid nanoparticle (LNP) preparation prepared by the ionizable lipid compound disclosed in the present application can effectively encapsulate mRNA and maintain the structural integrity of mRNA. The prepared ionizable lipid compound, distearoylphosphatidylcholine (DSPC, purchased from Nippon Seika Co., Ltd., Catalog No.: S01005), cholesterol (purchased from Nippon Seika Co., Ltd., Catalog No.: O01001) and dimyristoylglycerol-polyethylene glycol 2000 (DMG-PEG2000, purchased from Guobang Pharmaceutical, Catalog No.: O02005) were dissolved in ethanol (manufacturer: Nanjing Chemical Reagent Co., Ltd., purity 99.6%) solution, and then mixed according to a certain molar ratio to prepare a mixed lipid ethanol solution, wherein the total lipid concentration was 12.5 mM (the unit of measurement "M" in this application refers to mol / L). Firefly luciferase (Fluc) mRNA was diluted in 25 mM sodium acetate buffer at pH 5.0 to obtain an mRNA solution. By using a microfluidic device, the flow rate was controlled at 12 mL / min, the volume ratio of the mixed lipid ethanol solution to the mRNA solution prepared in the previous step was controlled to be 1:3, and lipid nanoparticles were prepared according to the nitrogen-phosphorus ratio of the ionizable lipid to the mRNA of 3 to 15:1. The ethanol was removed by dialysis against 20 mM Tris acetate for 12 to 24 hours. Finally, the LNP solution was filtered through a sterile filter with a pore size of 0.22 μm (manufacturer: Millex, product number: SLGPR33RB) and concentrated by ultrafiltration (manufacturer: Amicon-Ultra, molecular weight cutoff: 10 kDa) to obtain the LNP preparation obtained by encapsulating Fluc mRNA with the ionizable lipid described in this application and DSPC, cholesterol and DMG-PEG2000. The particle size and polydispersity index (PDI) of each LNP formulation were determined using dynamic light scattering using a Malvern Zetasizer Ultra instrument (Manufacturer: Malvern). The encapsulation efficiency of the LNPs was determined using the Quant-it Ribogreen RNA Quantification Kit (Manufacturer: ThermoFisher Scientific, Catalog No.: R11490).

[1897] Table 1

[1898]

[1899]

[1900] In the art, a PDI of less than 0.3 indicates relatively uniform nanoparticle size within the LNP formulation. Encapsulation efficiency is used to determine whether the LNP can effectively encapsulate mRNA, with an encapsulation efficiency above 70% indicating effective mRNA encapsulation. The LNPs prepared in this application maintained a particle size of 60 to 100 nm, a PDI of less than 0.2, and an encapsulation efficiency exceeding 90%.

[1901] Example 2 In vivo animal studies of LNP preparations

[1902] In this example, the LNPs prepared in Example 1 were injected into 6- to 8-week-old female Balb / C mice (Weitonglihua) via tail vein or lower limb intramuscular injection at a dose of 5 μg / mouse (n=3, i.e., 3 mice were injected and tested per group, and the data presented are the mean values ​​for each group). D-luciferin potassium salt was injected intraperitoneally at specific time points after administration (4 hours, 24 hours, and 48 hours in this example). Luminescence was then detected using an IVIS Spectrum small animal in vivo imager (manufacturer: PerkinElmer). The total luminescence intensity of the expression sites in the mice (e.g., liver, lower limb administration sites, etc.) was calculated. The higher the luminescence intensity, the higher the luciferase expression, i.e., the better the expression of the corresponding LNP formulation in the mice. The total luminescence intensity was measured by bioluminescence imaging, and the luminescence intensity data of the luminescent sites were collected 6 to 15 minutes (min) after the intraperitoneal injection of D-luciferin potassium salt. The total luminescence intensity of the expression areas in the mice was calculated using Living Image software (manufacturer: PerkinElmer). Normally, the total luminescence intensity reading of mice not treated with drugs is on the order of 10 5 .

[1903] Referring to the above-mentioned in vivo mouse experimental method, the LNP preparation in Example 1 was injected into 6-8 week old female Balb / C mice via the tail vein at a dose of 5 μg / mouse. The total luminescence intensity of the liver area was counted. The test results are shown in FIG. Figure 1 and Table 2. The LNP preparations tested in this example all showed strong expression in mice, with AUC ranging from 10 9 ~10 12 , indicating that the LNP preparations corresponding to the ionizable lipids described in the preparation examples can effectively deliver mRNA into the body and express it. It can be seen that the LNP preparations prepared in this example all have strong expression in mice.

[1904] Table 2 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1905] Group Liver AUC (p / s*hour) Compound 2 9.30E+11 Compound 3 3.00E+12 Compound 6 4.93E+12

[1906] Similarly, the LNP preparation prepared in Example 1 was injected intramuscularly (into the right lower calf) into female Balb / C mice aged 6 to 8 weeks at a dose of 5 μg / mouse. The total luminescence intensity of the liver region and the administration muscle site under this administration method was counted and the corresponding 4-48 h expression kinetic area under the curve (AUC) was calculated. The test results are shown in Figure 2 、 Figure 3 and Table 3. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1907] Table 3 Area under the expression kinetic curve (AUC) of liver region and administration site from 4 to 48 h

[1908]

[1909]

[1910] Example 3 Preparation and Detection of Lipid Nanoparticles (LNP)

[1911] In this example, Compound 8 and Compound 9 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared according to the molar ratios and nitrogen-phosphorus ratios in Table 4, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 4, the particle size of the LNP preparations prepared in this example ranged from 60 to 110 nm, the PDI was less than 0.2, and the encapsulation efficiency was greater than 90%.

[1912] Table 4

[1913]

[1914] Example 4

[1915] In vivo animal studies of LNP formulations

[1916] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 3 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the living mice was counted. The test results are shown in FIG. Figure 4 and Table 5. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1917] Table 5 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 h

[1918] Group Liver AUC (p / s*hour) Compound 8 1.98E+12 Compound 9 1.50E+12

[1919] Similarly, the LNP preparation prepared in Example 3 was injected intramuscularly (into the right lower calf) into female Balb / C mice aged 6 to 8 weeks at a dose of 5 μg / mouse. The total luminescence intensity of the liver region and the administration muscle site under this administration method was counted and the corresponding 4-48 h expression kinetic area under the curve (AUC) was calculated. The test results are shown in Figure 5 、 Figure 6 and Table 6. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1920] Table 6 Area under the expression kinetic curve (AUC) of liver region and administration site from 4 to 48 h

[1921] Group AUC at the administration site (p / s×hours) Liver AUC (p / s×hours) Compound 8 1.30E+11 4.22E+10 Compound 9 1.12E+11 3.45E+10

[1922] Example 5 Atomization stability test

[1923] The LNP preparations prepared in Example 1 and Example 3 were atomized by an aerosol needle (Shanghai Yuyan Scientific Instrument Co., Ltd.) (the molar percentage content and nitrogen-phosphorus ratio of the same group name representing the preparation were the same as in Examples 1 and 3). The physical and chemical characteristics of the samples before and after aerosolization were tested, and the results are shown in Table 7. As can be seen, after aerosolization, the particle size and PDI of each sample increased, and the encapsulation efficiency decreased. It is generally believed in the art that, using the determination method described in this application, the optimal standard for the stability of the preparation after aerosolization is: PDI <0.3, encapsulation efficiency >85%. The stability of the preparations of Compounds 3, Compound 6, Compound 8, and Compound 9 after aerosolization is excellent, and it is expected to improve their stability and efficacy when developed as aerosols.

[1924] Table 7

[1925]

[1926] Example 6 Preparation and Detection of Lipid Nanoparticles (LNP)

[1927] In this example, compound 5 and compound 28 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 8, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 8, the particle size of the LNP preparations prepared in this example was between 50 and 100 nm, the PDI was less than 0.2, and the encapsulation efficiency was greater than 90%.

[1928] Table 8

[1929]

[1930] Example 7 In vivo animal studies of LNP preparations

[1931] Referring to the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 6 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. Figure 7 As shown in Table 9, the LNP preparations prepared in this example were strongly expressed in mice. In terms of overall expression trend, compound 5 and compound 28 were slightly better than SM102.

[1932] Table 9 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1933] Group Liver AUC (p / s*hour) SM102 2.73E+12 Compound 5 4.00E+12 Compound 28 3.81E+12

[1934] Similarly, the LNP preparation prepared in Example 6 was injected intramuscularly (into the right lower calf) into female Balb / C mice aged 6 to 8 weeks at a dose of 5 μg / mouse. The total luminescence intensity of the liver region and the administration muscle site under this administration method was counted and the corresponding 4-48 h expression kinetic area under the curve (AUC) was calculated. The test results are shown in Figure 8 、 Figure 9 and Table 10. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1935] Table 10 Area under the expression kinetic curve (AUC) of liver region and administration site 4-48h

[1936] Group AUC at the administration site (p / s×hours) Liver AUC (p / s×hours) Compound 5 1.60E+11 1.35E+11 Compound 28 1.47E+11 1.07E+11

[1937] Example 8 Preparation and Detection of Lipid Nanoparticles (LNP)

[1938] In this example, compounds 23 to 27, compound 29, and compound 34 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared in the manner of Example 1 according to the molar ratios and nitrogen-phosphorus ratios in Table 11. The mRNA of this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 11, the particle size of the LNP preparations prepared in this example was between 60 and 100 nm, the PDI was less than 0.3, and the encapsulation efficiency was higher than 90%.

[1939] Table 11

[1940]

[1941]

[1942] Example 9 In vivo animal studies of LNP preparations

[1943] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 8 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 10 and Table 12. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1944] Table 12 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1945] Group Liver AUC (p / s*hour) Compound 23 4.22E+12 Compound 24 5.37E+12 Compound 25 3.02E+12 Compound 26 7.92E+12 Compound 29 5.12E+12 Compound 34 4.43E+12

[1946] Similarly, the LNP preparation prepared in Example 8 was injected intramuscularly (into the right lower calf) into female Balb / C mice aged 6 to 8 weeks at a dose of 5 μg / mouse. The total luminescence intensity of the liver region and the administration muscle site under this administration method was counted and the corresponding 4-48 h expression kinetic area under the curve (AUC) was calculated. The test results are shown in Figure 11 、 Figure 12 and Table 13. As can be seen, the LNP preparations prepared in this example all showed strong expression in mice. In terms of overall expression trend, compounds 23-26, compound 29, and compound 34 were slightly better than SM102, and compound 27 was better than SM102 in local administration.

[1947] Table 13 Area under the expression kinetic curve (AUC) of liver region and administration site 4-48h

[1948] Group AUC at the administration site (p / s×hours) Liver AUC (p / s×hours) SM102 1.45E+11 7.20E+10 Compound 23 3.74E+11 2.08E+11 Compound 24 3.07E+11 1.62E+11 Compound 25 3.41E+11 8.68E+10 Compound 26 3.59E+11 1.07E+11 Compound 27 2.02E+11 2.82E+10 Compound 29 2.70E+11 8.88E+10 Compound 34 2.27E+11 1.44E+11

[1949] Example 10 Atomization stability test

[1950] The LNP preparations prepared in Examples 6 and 8 were atomized using an aerosol needle (Shanghai Yuyan Scientific Instrument Co., Ltd.) (the same group names represent preparations with the same molar percentage content and nitrogen-phosphorus ratio as in Examples 6 and 8). The physical and chemical characteristics of the samples before and after atomization were tested. The results are shown in Table 14.

[1951] Table 14

[1952]

[1953] As can be seen, after nebulization, all samples showed an increase in particle size and PDI, while the encapsulation efficiency decreased. It is generally accepted in the art that, using the assay method described herein, the optimal stability criteria for a nebulized formulation are: PDI < 0.3 and encapsulation efficiency > 85%. Compound 25 exhibits excellent stability after nebulization, potentially improving its stability and efficacy when developed as an aerosol formulation.

[1954] Example 11 Preparation and Detection of Lipid Nanoparticles (LNP)

[1955] In this example, compound 4 was selected as the ionizable lipid, and an LNP preparation (encapsulating Fluc mRNA) was prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 15, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 15, the particle size of the LNP preparations prepared in this example was between 60 and 70 nm, the PDI was less than 0.1, and the encapsulation efficiency was greater than 90%.

[1956] Table 15

[1957]

[1958] Example 12 In vivo animal study of LNP preparations

[1959] Referring to the in vivo mouse testing method described in Example 2, the LNP formulation prepared in Example 11 was injected into 6- to 8-week-old female Balb / C mice via the tail vein at a dose of 5 μg / mouse. The total luminescence intensity in the liver of the mice was measured. The test results are shown in Table 16. The LNP formulation prepared in this example showed strong expression in the mice.

[1960] Table 16 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1961] Group Liver AUC (p / s*hour) Compound 4 3.56E+12

[1962] Example 13 Preparation and Detection of Lipid Nanoparticles (LNP)

[1963] This example selects compound 1, compound 7, compound 21, compound 32, compound 48, compound 55 and compound 57 as ionizable lipids, and prepares LNP preparations (encapsulating Fluc mRNA) in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 17. The mRNA of this example is diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 is used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this example are measured. As shown in Table 17, the particle size of the LNP preparations prepared in this example is between 50 and 80 nm, the PDI is less than 0.15, and the encapsulation efficiency is higher than 90%.

[1964] Table 17

[1965]

[1966] Example 14 In vivo animal study of LNP preparations

[1967] Referring to the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 13 was injected into 6-8 week old female Balb / C mice via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the mouse liver was counted. Figure 13 As shown in Table 18, the LNP preparations prepared in this example were strongly expressed in mice.

[1968] Table 18 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1969]

[1970]

[1971] Example 15 Preparation and Detection of Lipid Nanoparticles (LNP)

[1972] In this example, Compound 2, Compound 3, and Compound 6 were selected as ionizable lipids, and LNP preparations (encapsulating VZV mRNA) were prepared according to the molar ratios and nitrogen-phosphorus ratios in Table 19, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 19, the particle size of the LNP preparations prepared in this example ranged from 60 to 80 nm, the PDI was less than 0.1, and the encapsulation efficiency was greater than 90%.

[1973] Table 19

[1974]

[1975] Example 16 In vivo animal study of LNP preparations

[1976] Referring to the in vivo mouse test method of Example 2, the VZV-LNP reagent prepared in Example 15 was injected into female BABL / c mice aged 6 to 8 weeks through the tail vein at a dose of 60 μg / mouse, and whole blood of the mice was collected 3 hours after the injection. Serum was separated from the whole blood by centrifugation at 4°C and 2000g for 10 minutes and stored at -80°C for analysis. According to the manufacturer's instructions, enzyme-linked immunosorbent assay (ELISA) analysis was performed using Glycoprotein E (VZV) Standard (Bipsys, product number RAS102-C02) and Biotin-Anti-Glycoprotein E (VZV) Antibody (Bipsys, product number RAS102-C03) to measure the VZV antigen concentration. The test results are shown in Figure 14 and Table 20 (VZV antigen concentration in serum (n=5), statistical analysis by ANOVA, ****p<0.0001, *****p<0.0001, compared with the placebo group). Compared with the placebo group, the VZV antigen concentrations of animals in the SM102, Compound 2, Compound 3, and Compound 6 groups were significantly increased, and the VZV antigen concentration in the Compound 6 group was superior to that in the SM102 group.

[1977] Table 20 VZV antigen content in BABL / c mouse serum 3 hours after intravenous administration

[1978]

[1979]

[1980] Example 17 Preparation and Detection of Lipid Nanoparticles (LNP)

[1981] This example selects compound 11, compound 19-20, compound 22, compound 30-31, compound 52, compound 58 and compound 60 as ionizable lipids, and prepares LNP preparations (encapsulating Fluc mRNA) in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 21. The mRNA of this example is diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 is used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this example are measured. As shown in Table 21, the particle size of the LNP preparations prepared in this example is between 50 and 100 nm, the PDI is less than 0.2, and the encapsulation efficiency is higher than 90%.

[1982] Table 21

[1983]

[1984] Example 18 In vivo animal study of LNP preparations

[1985] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 22 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. Figure 15 As shown in Table 22, the LNP preparations prepared in this example were strongly expressed in mice.

[1986] Table 22 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1987]

[1988]

[1989] Example 19 Preparation and Detection of Lipid Nanoparticles (LNP)

[1990] This example selects compound 33, compound 35, compound 36, compound 37, compound 47, compound 49, compound 50, and compounds 62-68 as ionizable lipids, and prepares LNP preparations (encapsulating Fluc mRNA) in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 23. The mRNA of this example is diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 is used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example are measured. As shown in Table 23, the particle size of the LNP preparations prepared in this example is between 50 and 115 nm, the PDI is less than 0.20, and the encapsulation efficiency is higher than 90%.

[1991] Table 23

[1992]

[1993]

[1994] Example 20 In vivo animal studies of LNP preparations

[1995] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 19 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. Figure 16 As shown in Table 24, the LNP preparations prepared in this example were strongly expressed in mice.

[1996] Table 24 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1997] Group Liver AUC (p / s*hour) Compound 33 4.39E+12 Compound 35 1.56E+12 Compound 36 2.87E+10 Compound 37 1.88E+12 Compound 47 1.25E+12 Compound 49 2.63E+12 Compound 50 1.05E+12 Compound 62 1.82E+11 Compound 63 3.81E+12 Compound 64 2.97E+12 Compound 65 3.92E+12 Compound 66 2.32E+12 Compound 67 4.02E+12 Compound 68 9.62E+11

[1998] Example 21 Preparation and Detection of Lipid Nanoparticles (LNP)

[1999] In this example, compound 6 was selected as the ionizable lipid, and an LNP preparation (encapsulating Fluc mRNA) was prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 21, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 25, the particle size of the LNP preparations prepared in this example was between 50 and 100 nm, the PDI was less than 0.3, and the encapsulation efficiency was greater than 90%.

[2000] Table 25

[2001]

[2002]

[2003] Example 22 In vivo animal study of LNP preparations

[2004] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 21 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. Figure 17 As shown in Table 26, the LNP preparations prepared in this example were strongly expressed in mice.

[2005] Table 26 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[2006] Group Liver AUC (p / s*hour) LNP1 1.27E+12 LNP2 2.26E+12 LNP3 2.80E+12 LNP4 4.45E+12 LNP5 3.59E+12 LNP6 2.17E+12 LNP7 8.26E+12 LNP8 7.62E+12 LNP9 1.23E+12 LNP10 3.08E+11 LNP11 4.61E+12 LNP12 5.13E+12 LNP13 2.08E+12 LNP14 6.88E+11

[2007] Example 23 Preparation and Detection of Lipid Nanoparticles (LNP)

[2008] In this example, compound 6 was selected as the ionizable lipid, and an LNP preparation (encapsulating Fluc mRNA) was prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 26, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 27, the particle size of the LNP preparations prepared in this example was between 50 and 70 nm, the PDI was less than 0.10, and the encapsulation efficiency was greater than 95%.

[2009] Table 27

[2010]

[2011] Example 24 In vivo animal study of LNP preparations

[2012] Referring to the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 23 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. Figure 18 As shown in Table 28, the LNP preparations prepared in this example were strongly expressed in mice.

[2013] Table 28 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[2014] Group Liver AUC (p / s*hour) NP3 2.82E+12 NP4 3.41E+12 NP5 4.30E+12 NP6 4.90E+12 NP7 5.20E+12 NP8 6.35E+12 NP9 5.95E+12

[2015] Example 25 Preparation and Detection of Lipid Nanoparticles (LNP)

[2016] In this example, compound 10, compound 12-17, and compound 61 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 28, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 29, the particle size of the LNP preparations prepared in this example was between 50 and 140 nm, the PDI was less than 0.3, and the encapsulation efficiency was higher than 90%.

[2017] Table 29

[2018]

[2019]

[2020] Example 26 In vivo animal study of LNP preparations

[2021] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 25 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. Figure 19 As shown in Table 30, the LNP preparations prepared in this example were strongly expressed in mice.

[2022] Table 30 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[2023] Group Liver AUC (p / s*hour) Compound 10 1.86E+11 Compound 12 2.35E+12 Compound 13 5.53E+10 Compound 14 1.75E+12 Compound 15 8.35E+11 Compound 16 1.50E+12 Compound 17 1.19E+12 Compound 61 2.90E+10 .

Claims

1. A compound I or a pharmaceutically acceptable salt thereof, in, R 1 is a C substituted with one or more hydroxyl groups 1-6 alkyl; X and Y are independently C 1-12 Alkylene; Z 1 and Z 2 Independently W 1 and W 2 Independently a chemical bond or C 1-6 Alkylene; R 2 for R 2a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-6 Alkyl; R 2b C 1-6 Alkylene, R 2c C 1-15 alkyl; R 3 C 1-15 Alkyl or R 3a C 1-6 Alkylene, R 3b C 1-15 alkyl; R 4 H, C 1-15 alkyl, or -C 3-10 Cycloalkyl-C 1-15 Alkyl; R 4a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa ; R 4aa H or C 1-6 alkyl; R 5 C 1-15 Alkyl or R 5a C 1-6 Alkylene, R 5b C 1-15 alkyl.

2. The compound I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It satisfies any of the following options: Solution (1) Z 1 and Z 2 Independently R 2 for R 2a C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-6 alkyl; Solution (2) R 1 is a C substituted with one or more hydroxyl groups 1-6 alkyl; X and Y are independently C 1-12 Alkylene; Z 1 for *Denotes the same as W 1 connect; Z 2 for *Denotes the same as W 2 connect; W 1 and W 2 Independently a chemical bond or C 1-6 Alkylene; R 2 for R 2a C 1-15 Alkyl or C 1-15 alkenyl; R 3 C 1-15 alkyl; R 4 C 1-15 Alkyl or R 4a C 1-15 Alkyl or C 1-15 alkenyl; R 5 C 1-15 alkyl; Solution (3) R 1 is a C substituted with one or more hydroxyl groups 1-6 alkyl; X and Y are independently C 1-12 Alkylene; Z 1 for *Denotes the same as W 1 connect; Z 2 for *Denotes the same as W 2 connect; W 1 and W 2 Independently a chemical bond or C 1-6 Alkylene; R 2 for R 2a C 1-15 Alkyl or C 1-15 alkenyl; R 3 C 1-15 Alkyl or R 3a C 1-6 Alkylene, R 3b C 1-15 Alkyl; R 4 C 1-15 Alkyl or R 4a C 1-15 Alkyl or C 1-15 alkenyl; R 5 C 1-15 Alkyl or R 5a C 1-6 Alkylene, R 5b C 1-15 alkyl.

3. The compound I or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: It meets one or more of the following conditions: (1)R 1 , R 2aa and R 4aa wherein the C 1-6 Alkyl and C 1-6 The "C 1-6 "alkyl" are each independently C 1-4 Alkyl, such as methyl, ethyl, n-propyl or n-butyl; (2) X, Y, R 2a and R 4a In the C 1-12 Alkylene, -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa , -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa "C 1-12 "Alkylene" is each independently C 3-7 Alkylene, e.g. (3)W 1 , W 2 , R 3a and R 5a In the C 1-6 The alkylene groups are each independently methylene, -CH2CH2-, (4)R 2a , R 3 , R 3b , R 4 , R 4a , R 5 and R 5b In the C 1-15 Alkyl, -C 3-10 Cycloalkyl-C 1-15 The "C 1-15 "Alkyl" is independently C 3-12 Alkyl, such as n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, (5)R 2a and R 4a In the C 1-15 The alkenyl groups are independently C 3-10 Alkenyl, e.g. (6)R 2a and R 4a In the -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 2aa and -C 1-12 Alkylene-C 3-6 Cycloalkyl-R 4aa "C 3-6 "Cycloalkyl" is independently cyclopropyl or cyclobutyl; (7)R 4 In the -C 3-10 Cycloalkyl-C 1-15 The "C 3-10 "Cycloalkyl" is C 3-6 Cycloalkyl, for example cyclohexyl.

4. The compound I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It meets one or more of the following conditions: (1)R 2b In the C 1-6 Alkylene is C 1-3 Alkylene groups, such as methylene; (2)R 2c In the C 1-15 Alkyl is C 3-12 Alkyl, such as n-pentyl or n-heptyl; (3)R 2a In the C 1-15 Alkyl is C 3-10 Alkyl, such as n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, (4) Z 2 for *Denotes the same as W 1 connect; (5)R 2 for R 2a C 3-10 Alkyl, C 3-10 Alkenyl or -C 3-7 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-3 Alkyl; R 2b C 1-3 Alkylene, R 2c C 3-10 alkyl; (6)R 2b C 1-3 Alkylene, such as methylene; R 2c C 3-10 Alkyl groups, e.g. (7)R 2a for (8)R 2 for (9)R 3 for (10)R 3b for (11) for (12)R 4a for (13)R 4 for (14)R 5b is n-octyl; (15)R 5 for (16) for 5. The compound I or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: It meets one or more of the following conditions: (1)R 1 C is substituted by a hydroxyl group 1-6 alkyl; (2) X and Y are independently C 3-7 Alkylene; (3) Z 1 for *Denotes the same as W 1 connect; (4) Z 2 for *Denotes the same as W 2 connect; (5)W 1 and W 2 are independently a chemical bond, a methylene group, -CH2CH2-, (6)R 2a C 3-10 Alkyl, C 3-10 Alkenyl or -C 3-7 Alkylene-C 3-6 Cycloalkyl-R 2aa , where R 2aa H or C 1-3 Alkyl; preferably, R 2a is n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, (7)R 3 C 4-12 Alkyl or R 3a C 1-3 Alkylene, R 3b C 6-12 Alkyl; preferably, R 3a R is methylene; 3b is n-heptyl; (8)R 4 H, C 3-10 alkyl, or -C 3-6 Cycloalkyl-C 3-10 Alkyl; R 4a C 3-10 Alkyl, C 3-10 Alkenyl or -C 3-7 Alkylene-C 3-6 Cycloalkyl-R 4aa ; R 4aa H or C 1-3 Alkyl; preferably, R 4a is n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, (9)R 5 C 6-10 Alkyl or R 5a C 1-3 Alkylene, R 5b C 6-10 Alkyl; preferably, R 5a R is methylene; 5b It is n-heptyl.

6. Compound I or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: It meets one or more of the following conditions: (1)R 1 for (2) X and Y are independently (3)R 2 for (4)R 3 for (5)R 4 For H. (6)R 5 for (7) for (8) for 7. The compound I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound I satisfies any of the following schemes: The compound I described in scheme (1) has the following structure: Among them, R 1 , R 2 , R 3 , R 4 , R 5 , W 1 , W 2 , X and Y are defined as described in any one of claims 1 to 6; preferably, the compound I has the following structure: Among them, R 1 , R 2a , R 3 , R 4a , R 5 , W 1 , W 2 , X and Y are defined as any one of claims 1-6; the compound I described in scheme (2) is the following structure: Among them, R 1 , R 2 , R 3 , R 4 , R 5 , W 1 , W 2 , X and Y are defined as any one of claims 1-6; the compound I described in scheme (3) is any of the following structures: Among them, R 1 , R 2 , R 3 , R 4 , R 5 , W 1 , W 2 , Z 1 , Z 2 , X and Y are as defined in any one of claims 1-6.

8. The compound I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The compound I is any of the following structures:

9. A lipid carrier comprising a substance Z, wherein the substance Z is the compound I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8; Preferably, the lipid carrier satisfies one or more of the following conditions: (1) The lipid carrier further comprises a diluent, such as a phosphate buffer or a Tris buffer; (2) The lipid carrier further comprises a phospholipid, which is a phospholipid molecule having an electrically charged polar end and a non-polar fatty chain end, such as distearoylphosphatidylcholine, dimyristoylphosphocholine, dioleoylphosphocholine, palmitoylphosphocholine, 1,2-distearoylphosphocholine, heneicosanoylphosphocholine or palmitoylphosphocholine; (3) The lipid carrier further comprises PEG lipid, wherein the PEG lipid is a lipid molecule having a polyethylene glycol-modified hydrophilic end, such as PEG-modified dimyristoylglycerol; (4) the lipid carrier further comprises a sterol, wherein the sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol; (5) In the lipid carrier, the molar ratio of the substance Z to the sterol is 0.5-5:1, preferably 0.5-3:1, for example 1.3:1; (6) In the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 2-8:1, for example 3-6:1; (7) In the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 1-10:1, for example 5:1; (8) In the lipid carrier, the molar ratio of the substance Z to the PEG lipid is 10-100:1, preferably 10-50:1, for example 33.3:1; (9) In the lipid carrier, the molar content of the substance Z is 30 mol% to 70 mol%, for example 50 mol%; (10) In the lipid carrier, the molar content of the phospholipid is 5 mol% to 20 mol%, for example, 10 mol%; (11) In the lipid carrier, the molar content of the sterol is 20 mol% to 60 mol%, for example, 38.5 mol%; (12) In the lipid carrier, the molar content of the PEG lipid is about 0.2 mol% to 5 mol%, for example, 1.5% mol; (13) In the lipid carrier, the lipid carrier is composed of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.

10. The lipid carrier according to claim 9, characterized in that It meets one or more of the following conditions: (1) In the lipid carrier, the molar content of the substance Z is 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%; (2) In the lipid carrier, the molar content of the phospholipid is 0 mol% to 30 mol%, for example, 0 mol%, 5 mol%, 10 mol%, 15 mol% or 20 mol%; (3) In the lipid carrier, the molar content of the sterol is 28.5 mol%, 33.5 mol%, 37.0 mol%, 38.0 mol%, 38.5 mol%, 39.0 mol%, 39.5 mol%, 43.5 mol%, 48.5 mol% or 53.5 mol%; (4) In the lipid carrier, the molar content of the PEG lipid is 0.5% mol, 1.0% mol, 1.5% mol, 2.0% mol or 3.0% mol; (5) The lipid carrier is any of the following formulations: 。 11. Use of the compound I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, or the lipid carrier according to claim 9 in the preparation of a nucleic acid preventive agent and / or therapeutic agent delivery vector; The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid, double-stranded DNA, small interfering RNA, asymmetric double-stranded small interfering RNA, microRNA, small hairpin RNA, circular RNA, transfer RNA or messenger RNA, preferably mRNA, such as firefly luciferase mRNA, SARS-CoV-2 spike protein mRNA, herpes zoster virus mRNA, respiratory syncytial virus mRNA or influenza virus mRNA.

12. A lipid nanoparticle comprising a nucleic acid preventive and / or therapeutic agent and the lipid carrier according to claim 9; The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid, double-stranded DNA, small interfering RNA, asymmetric double-stranded small interfering RNA, microRNA, small hairpin RNA, circular RNA, transfer RNA or messenger RNA, preferably mRNA, such as firefly luciferase mRNA, SARS-CoV-2 spike protein mRNA, herpes zoster virus mRNA, respiratory syncytial virus mRNA or influenza virus mRNA.

13. The lipid nanoparticle according to claim 12, characterized in that It meets one or more of the following conditions: (1) The nitrogen-to-phosphorus ratio in the lipid nanoparticles is (2-30):1, preferably (2-20):1, more preferably (3-20):1, for example (3-16):1; (2) The particle size of the lipid nanoparticles is 10-200 nm, preferably 40-150 nm, more preferably 50-130 nm, for example 57.29 nm, 61.55 nm, 69.72 nm, 70.08 nm, 76.03 nm, 89.89 nm, 97.55 nm, 107.37 nm or 119.10 nm; (3) the polydispersity index of the lipid nanoparticles is 0.001-0.3, for example, 0.035, 0.074, 0.090, 0.097, 0.118, 0.166 or 0.269; (4) the encapsulation efficiency of the lipid nanoparticles is 90%-100%, for example, 96.73%, 98.07%, 96.90%, 97.71%, 96.36%, 95.02%, 92.94%, 94.44%, 95.23% or 94.63%; (5) In the lipid nanoparticles, the lipid carrier encapsulates the nucleic acid preventive agent and / or therapeutic agent.

14. The lipid nanoparticle according to claim 12, characterized in that It meets one or more of the following conditions: (1) In the lipid nanoparticles, the nitrogen-to-phosphorus ratio is 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or 9:1; (2) the particle size of the lipid nanoparticles is 57.29 nm, 61.55 nm, 69.72 nm, 70.08 nm, 76.03 nm, 89.89 nm, 97.55 nm, 107.37 nm, 119.10 nm, 63.36 mm, 56.53 mm, 71.25 mm, 57.66 mm, 60.92 mm, 55.68 mm, 56.19 mm, 55.52 mm, 67.96 mm, 69.51 mm or 79.13 mm; (3) the polydispersity index of the lipid nanoparticles is 0.035, 0.074, 0.090, 0.097, 0.118, 0.166, 0.269, 0.065, 0.074, 0.062, 0.106, 0.089, 0.108, 0.090, 0.078, 0.085, 0.063 or 0.084; (4) The encapsulation efficiency of the lipid nanoparticles is 96.73%, 98.07%, 96.90%, 97.71%, 96.36%, 95.02%, 92.94%, 94.44%, 95.23%, 94.63%, 95.83%, 94.95%, 96.00%, 96.62%, 94.28%, 96.56%, 96.04%, 95.94%, 98.33%, 95.38% or 96.78%.

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