Nitrogen-containing chain compound, preparation method therefor, composition containing same, and use thereof

Abstract

Disclosed in the present invention are a nitrogen-containing chain compound, a preparation method therefor, a composition containing same, and the use thereof. Specifically disclosed in the present invention is a compound of formula I or a pharmaceutically acceptable salt thereof, which has the following advantage: an LNP formulation prepared therefrom exhibits a relatively uniform nanoparticle size, a high encapsulation efficiency, and a high in-vivo expression activity.

Description

Nitrogen-containing chain compounds, preparation methods, compositions containing them, and applications.

[0001] This application claims priority to Chinese patent application 2024115676538, filed November 4, 2024; Chinese patent application 2025102398736, filed February 28, 2025; and Chinese patent application 2025107130220, filed May 29, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This invention relates to a nitrogen-containing chain compound, a method for its preparation, compositions comprising the compound, and its applications. Background Technology

[0003] Nucleic acid drugs represent an important direction in current basic and applied research. They can be used for the prevention and / or treatment of viral and bacterial infectious diseases, tumors, metabolic diseases, etc., and their production costs are lower and the cycle shorter, facilitating the rapid development of personalized medicines. However, nucleic acids are negatively charged macromolecules, making them difficult to penetrate cell membranes, and they also exhibit poor stability. Developing various nucleic acid packaging and delivery systems can, to some extent, overcome the instability of nucleic acid drugs and improve their delivery efficiency.

[0004] Lipid nanoparticles have been shown to be effective carriers 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 amounts of components within lipid nanoparticles is of great significance for improving the efficacy of nucleic acid drugs for prevention and treatment, especially for lipid compounds and related methods and compositions for delivering RNA preventative agents and / or therapeutic agents.

[0005] Given the importance of ionizable lipid compounds that can be used to deliver nucleic acid drugs, there is an urgent need to develop novel ionizable lipid compounds. Summary of the Invention

[0006] This invention aims to provide a novel ionizable lipid compound for delivering nucleic acid drugs, increasing the variety of ionizable lipid compounds and the selection of delivery carriers for nucleic acid prophylaxis and / or therapeutic agents. To address the above technical problems, this invention provides a nitrogen-containing chain compound, a preparation method, compositions comprising the same, and applications. LNP formulations prepared using the nitrogen-containing chain compound of this invention exhibit relatively uniform nanoparticle size, high encapsulation efficiency, and high in vivo expression activity.

[0007] This invention provides a compound of formula I or a pharmaceutically acceptable salt thereof.

[0008] Among them, R1 A hydroxyl group, or a C substituted with one or more hydroxyl groups. 1-6 alkyl;

[0009] Ring A is C 3-8 Cycloalkylene;

[0010] L 1 For chemical bonds or C 1-3 Alkylene;

[0011] X and Y are independently C 1-15 Alkylene;

[0012] Z 1 for -O-、-OZ 1c -C(=O)-O- or chemical bond;

[0013] Z 1a Z 1b and Z 1c Independently for C 1-6 Alkylene;

[0014] Z 2 for -O- or -OZ 2c -C(=O)-O-;

[0015] Z 2a Z 2b and Z 2c Independently for C 1-6 Alkylene;

[0016] W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene;

[0017] R 2 C 1-15 Alkyl, -R 2a -OC 1-15 alkyl,

[0018] R 2a For chemical bonds or C 1-6 Alkylene; R 2c and R 2d Independently for C 1-15 Alkyl or C 1-15 alkenyl;

[0019] R 2b C 1-15 Alkyl, C 1-15 alkenyl, -C 1-15 Alkylene-R 2ba -C1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 Alkyl; R 2ba It is a 3-10 membered heterocyclic alkyl group; wherein the heterocyclic alkyl group has 1-3 heteroatoms, and the heteroatoms are independently N, O or S;

[0020] R 3 For H, C 1-15 Alkyl, -R 3a -OC 1-15 alkyl,

[0021] R 3a For chemical bonds or C 1-6 Alkylene; R 3b C 1-6 Alkylene; R 3c R 3d and R 3e Independently for C 1-15 alkyl;

[0022] R 4 C 1-15 Alkyl, -R 4a -OC 1-15 alkyl,

[0023] R 4a For chemical bonds or C 1-6 Alkylene; R 4b C 1-6 Alkylene;

[0024] R 4c and R 4e Independently for C 1-15 Alkyl or C 1-15 alkenyl;

[0025] R 4d C 1-15 Alkyl, C 1-15 alkenyl, -C 1-15 Alkylene-R 4da -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 alkyl;

[0026] R 4da It is a 3-10 membered heterocyclic alkyl group; wherein the heterocyclic alkyl group has 1-3 heteroatoms, and the heteroatoms are independently N, O or S;

[0027] R 5For H, C 1-15 alkyl,

[0028] R 5a C 1-6 Alkylene; R 5b and R 5c Independently for C 1-15 alkyl.

[0029] This invention provides a compound of formula I or a pharmaceutically acceptable salt thereof.

[0030] Among them, R 1 A hydroxyl group, or a C substituted with one or more hydroxyl groups. 1-6 alkyl;

[0031] Ring A is C 3-8 Cycloalkylene;

[0032] L 1 For chemical bonds or C 1-3 Alkylene;

[0033] X and Y are independently C 1-15 Alkylene;

[0034] Z 1 for Or chemical bonds;

[0035] Z 1a and Z 1b Independently for C 1-6 Alkylene;

[0036] Z 2 for

[0037] Z 2a and Z 2b Independently for C 1-6 Alkylene;

[0038] W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene;

[0039] R 2 C 1-15 Alkyl, -R 2a -OC 1-15 alkyl,

[0040] R 2a For chemical bonds or C 1-6 Alkylene; R 2b R 2c and R2d Independently for C 1-15 Alkyl or C 1-15 alkenyl;

[0041] R 3 For H, C 1-15 Alkyl, -R 3a -OC 1-15 alkyl,

[0042] R 3a For chemical bonds or C 1-6 Alkylene; R 3b C 1-6 Alkylene; R 3c R 3d and R 3e Independently for C 1-15 alkyl;

[0043] R 4 C 1-15 Alkyl, -R 4a -OC 1-15 alkyl,

[0044] R 4a For chemical bonds or C 1-6 Alkylene; R 4b C 1-6 Alkylene;

[0045] R 4c R 4d and R 4e Independently for C 1-15 Alkyl or C 1-15 alkenyl;

[0046] R 5 For H, C 1-15 alkyl,

[0047] R 5a C 1-6 Alkylene; R 5b and R 5c Independently for C 1-15 alkyl.

[0048] In certain preferred embodiments of the present invention, certain groups of the compound of Formula I or its pharmaceutically acceptable salt are defined as follows, and groups not mentioned are as described in any embodiment of the present invention (hereinafter referred to as "in one embodiment of the present invention"). Other aspects of the present invention are referred to using the same abbreviations as this invention and will not be described further.

[0049] In one aspect of the present invention, R 1C is a hydroxyl group or a C substituted with a hydroxyl group. 1-3 alkyl.

[0050] In one aspect of the present invention, R 1 for

[0051] In one aspect of the present invention, R 1 for

[0052] In one embodiment of the present invention, ring A is C. 4-6 Cycloalkylene, for example

[0053] In one aspect of the present invention, for

[0054] Preferably, for

[0055] In one aspect of the present invention, for

[0056] In one aspect of the present invention, L 1 It is a chemical bond or a methylene group, preferably a chemical bond.

[0057] In one embodiment of the present invention, X and Y are independently C. 3-10 Alkylene.

[0058] In one embodiment of the present invention, X and Y are independently...

[0059] In one embodiment of the present invention, X and Y are independently...

[0060] In one embodiment of the present invention, X and Y are independently...

[0061] In one aspect of this invention, Z 1a and Z 1b Independently for C 1-3 Alkylene, for example

[0062] In one aspect of this invention, Z 1a and Z 1b Independently for C 1-3 alkylene, such as methylene or

[0063] In one aspect of this invention, Z 1c C 1-3 Alkylenes, such as methylene,

[0064] In one aspect of this invention, Z 1 for Or a chemical bond, * indicates the end connected to X.

[0065] In one aspect of this invention, Z 1 for -O- or * -OZ 1c -C(=O)-O-, * indicates the end connected to X.

[0066] In one aspect of this invention, Z 1 for -O-、 * -OZ 1c -C(=O)-O- or chemical bond, * indicates the end connected to X.

[0067] In one aspect of this invention, Z 1 for Or chemical bonds;

[0068] Preferably, Z 1 for Or a chemical bond, * indicates the end connected to X.

[0069] In one aspect of this invention, Z 1 for -O-、 * indicates the end connected to X;

[0070] Preferably, Z 1 for -O-、 * indicates the end connected to X.

[0071] In one aspect of this invention, Z 1 for -O-、 Or chemical bonds;

[0072] Preferably, Z 1 for -O-、 Or a chemical bond, * indicates the end connected to X.

[0073] In one aspect of this invention, Z 2a and Z 2b Independently for C 1-4 Alkylene, for example

[0074] In one aspect of this invention, Z 2a and Z 2b Independently for C 1-4 Alkylene, for example

[0075] In one aspect of this invention, Z 2c C 1-4 Alkylenes, such as methylene,

[0076] In one aspect of this invention, Z 2 for * indicates the end connected to Y.

[0077] In one aspect of this invention, Z 2 for -O- or * -OZ 2c -C(=O)-O-, * indicates the end connected to Y.

[0078] In one aspect of this invention, Z 2 for -O- or * -OZ 2c -C(=O)-O-, * indicates the end connected to Y.

[0079] In one aspect of this invention, Z 2 for

[0080] Preferably, Z 2 for * indicates the end connected to Y.

[0081] In one aspect of this invention, Z 2 for -O-、 * indicates the end connected to Y;

[0082] Preferably, Z 2 for -O-、 * indicates the end connected to Y.

[0083] In one aspect of this invention, Z2 for -O-、

[0084] Preferably, Z 2 for -O-、 * indicates the end connected to Y.

[0085] In one aspect of the present invention, W 1 and W 2 Independently for C 1-6 Alkylene, preferably C 1-3 Alkylene.

[0086] In one aspect of the present invention, W 1 and W 2 Independent of chemical bonds or C 1-3 Alkylene.

[0087] In one aspect of the present invention, W 1 and W 2 Independent of chemical bonds, methylene,

[0088] In one aspect of the present invention, R 2a For chemical bonds or C 1-3 Alkylene.

[0089] In one aspect of the present invention, R 2a It is a chemical bond or a methylene group.

[0090] In one aspect of the present invention, in the compound of formula I, R 2c R 2d R 2b R 4c R 4e and R 4d In, the C 1-15 The alkenyl group is independently C 2-15 Alkenyl group.

[0091] In one aspect of the present invention, in the compound of formula I, each "C" 1-15 "Alkenyl" is independently "C" 2-15 Alkenyl group.

[0092] In one aspect of the present invention, R 2b C 5-15 Alkyl or C 5-15 Alkenyl group.

[0093] In one aspect of the present invention, R 2b -C 3-15 Alkylene-R2ba -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 alkyl.

[0094] In one aspect of the present invention, R 2b C 3-15 Alkyl, C 5-15 alkenyl, -C 3-15 Alkylene-R 2ba -C 1-6 Alkylene-OC 3-8 Alkyl, -C 1-6 Alkylene-SC 3-8 alkyl.

[0095] In one aspect of the present invention, R 2ba It is a 5-8 membered heterocyclic alkyl group; the heterocyclic alkyl group preferably has 2 heteroatoms, and the heteroatoms are preferably S.

[0096] In one aspect of the present invention, R 2ba for

[0097] In one aspect of the present invention, R 2b for

[0098] In one aspect of the present invention, R 2b for

[0099] In one aspect of the present invention, R 2b for

[0100] In one aspect of the present invention, R 2c C 5-15 alkyl.

[0101] In one aspect of the present invention, R 2c for

[0102] In one aspect of the present invention, R 2d C 1-15 alkyl.

[0103] In one aspect of the present invention, R 2d for

[0104] In one aspect of the present invention, R 2 C 5-15 Alkyl, -R2a -OC 5-15 alkyl, R 2 Preferred is -R 2a -OC 1-15 alkyl,

[0105] In one aspect of the present invention, R 2 for

[0106] In one aspect of the present invention, R 2 for

[0107] In one aspect of the present invention, R 2 for

[0108] In one aspect of the present invention, R 3a C 1-3 Alkylene, such as methylene.

[0109] In one aspect of the present invention, R 3b C 1-3 Alkylene, such as methylene.

[0110] In one aspect of the present invention, R 3c C 5-15 Alkyl, for example

[0111] In one aspect of the present invention, R 3d C 5-15 Alkyl, for example

[0112] In one aspect of the present invention, R 3e C 1-15 Alkyl, for example

[0113] In one aspect of the present invention, R 3e C 5-15 Alkyl, for example

[0114] In one aspect of the present invention, R 3e C 5-15 Alkyl, for example

[0115] In one aspect of the present invention, R3 For H, C 5-15 Alkyl, -R 3a -OC 5-15 alkyl,

[0116] In one aspect of the present invention, R 3 For H,

[0117] In one aspect of the present invention, R 3 for

[0118] In one aspect of the present invention, R 3 For H,

[0119] In one aspect of the present invention, for

[0120] In one aspect of the present invention, for

[0121] In one aspect of the present invention, for

[0122] In one aspect of the present invention, R 4a It is a chemical bond.

[0123] In one aspect of the present invention, R 4b C 1-3 Alkylene, such as methylene.

[0124] In one aspect of the present invention, R 4c C 5-15 Alkyl, for example

[0125] In one aspect of the present invention, R 4d C 5-15 Alkyl or C 5-15 alkenyl, for example

[0126] In one aspect of the present invention, R 4d -C3-15 Alkylene-R 4da -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 alkyl.

[0127] In one aspect of the present invention, R 4d C 3-15 Alkyl, C 5-15 alkenyl, -C 3-15 Alkylene-R 4da -C 1-6 Alkylene-OC 3-8 Alkyl, -C 1-6 Alkylene-SC 3-8 alkyl.

[0128] In one aspect of the present invention, R 4da It is a 5-8 membered heterocyclic alkyl group; the heterocyclic alkyl group preferably has 2 heteroatoms, and the heteroatoms are preferably S.

[0129] In one aspect of the present invention, R 4da for

[0130] In one aspect of the present invention, R 4d for

[0131] In one aspect of the present invention, R 4d for

[0132] In one aspect of the present invention, R 4e C 5-15 Alkyl, for example

[0133] In one aspect of the present invention, R 4 C 5-15 Alkyl, -R 4a -OC 5-15 alkyl,

[0134] In one aspect of the present invention, R 4 for

[0135] In one aspect of the present invention, R 4 for

[0136] In one aspect of the present invention, R 4 for

[0137] In one aspect of the present invention, R 5a C 1-3 Alkylene, such as methylene.

[0138] In one aspect of the present invention, R 5b C 5-15 Alkyl, for example

[0139] In one aspect of the present invention, R 5c C 5-15 Alkyl, for example

[0140] In one aspect of the present invention, R 5c C 5-15 Alkyl, for example

[0141] In one aspect of the present invention, R 5c C 5-15 Alkyl, for example

[0142] In one aspect of the present invention, R 5 For H, C 5-15 alkyl,

[0143] In one aspect of the present invention, R 5 For H,

[0144] In one aspect of the present invention, R 5 for

[0145] In one aspect of the present invention, R 5 For H,

[0146] In one aspect of the present invention, for

[0147] In one aspect of the present invention, for

[0148] In one aspect of the present invention, for

[0149] In one aspect of the present invention, the compound of formula I has the following structure:

[0150] Among them, R 1 Ring A, L 1 X, Y, Z 1 Z 2 W 1 W 2 R 2b R 3 R 4 and R 5 The definition is as described in any of the previous schemes;

[0151] Preferably, the compound of formula I has the following structure:

[0152] Among them, R 1 Ring A, L 1 X, Y, Z 1 Z 2 W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any of the previous schemes;

[0153] in, Preferred W 1 and W 2 C is preferred 1-3 Alkylene;

[0154] More preferably, the compound of formula I has the following structure:

[0155] Among them, R 1 Rings A, X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any of the previous schemes;

[0156] Among them, ring A is preferred.

[0157] More preferably, the compound of formula I has the following structure:

[0158] Among them, X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in the previous scheme.

[0159] In one aspect of the present invention, the compound of formula I has the following structure:

[0160] Among them, R 1 X, Y, Z 1 Z 2 W 1 W 2 R 2 R 3 R 4 and R 5 The definition is as described in any of the previous schemes;

[0161] Preferably, R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl;

[0162] X and Y are independently C 3-10 Alkylene;

[0163] Z 1 for * indicates the end connected to X; Z 1a C 1-6 Alkylene;

[0164] Z 2 for * indicates the end connected to Y; Z 2a Independently for C 1-6 Alkylene;

[0165] W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene;

[0166] R 2 C 1-15 alkyl or R 2b C 1-15 alkyl;

[0167] R 3 C 1-15 alkyl;

[0168] R 4 C 1-15 alkyl or R 4d C 1-15 alkyl;

[0169] R 5 C 1-15 alkyl.

[0170] In one aspect of the present invention, the compound of formula I has the following structure:

[0171] Among them, R 1 X, Y, W 1 W 2 R 2 R 3 R 4 and R 5 The definition is as described in any of the previous schemes;

[0172] Preferably, R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl;

[0173] X and Y are independently C 3-10 Alkylene;

[0174] W 1 and W 2 Independently for C 1-6 Alkylene;

[0175] R 2 C 1-15 alkyl or R 2b C 1-15 alkyl;

[0176] R 3 C 1-15 alkyl;

[0177] R 4 C 1-15 alkyl or R 4d C 1-15 alkyl;

[0178] R 5 C 1-15 alkyl.

[0179] In one aspect of the present invention, the compound of formula I has the following structure:

[0180] Among them, R 1 X, Y, W1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any of the previous schemes;

[0181] Preferably,

[0182] R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl;

[0183] X and Y are independently C 3-10 Alkylene;

[0184] W 1 and W 2 Independently for C 1-6 Alkylene;

[0185] R 2b C 1-15 alkyl;

[0186] R 3 C 1-15 alkyl;

[0187] R 4d C 1-15 alkyl;

[0188] R 5 C 1-15 alkyl;

[0189] More preferably,

[0190] R 1 It is a hydroxyl group, or -CH2-OH;

[0191] X and Y are independently C 4-8 Alkylene, preferably C 5-7 Alkylene, for example

[0192] W 1 and W 2 Independently for C 1-3 Alkyl groups, such as methylene groups;

[0193] R 2b C 6-10 Alkyl group, preferably C 6-8 Alkyl, for example

[0194] R 3 C 1-15 Alkyl group, preferably C8-12 Alkyl, more preferably C 8-10 Alkyl, for example

[0195] R 4d C 1-15 Alkyl group, preferably C 4-9 Alkyl, more preferably C 5-8 Alkyl, for example

[0196] R 5 C 1-15 Alkyl group, preferably C 6-12 Alkyl, more preferably C 7-10 Alkyl, for example

[0197] In one aspect of the present invention, the compound of formula I has the following structure:

[0198] Among them, X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any of the previous schemes;

[0199] Preferably,

[0200] X and Y are independently C 5-7 Alkylene, for example

[0201] W 1 and W 2 Independently for C 1-3 Alkyl groups, such as methylene groups;

[0202] R 2b C 6-8 Alkyl, for example

[0203] R 3 C 8-10 Alkyl, for example

[0204] R 4d C 5-8 Alkyl, for example

[0205] R 5 C7-10 Alkyl, for example

[0206] In one aspect of the present invention, the compound of formula I has any of the following structures:

[0207] Among them, R 1 X, Y, Z 1 Z 2 W 1 W 2 R 2 R 3 R 4 and R 5 The definition is as described in any of the previous schemes;

[0208] Preferably, the structure of formula I-3A is as shown in I-3Aa:

[0209] Preferably, the structure of formula I-3B is as shown in I-3Ba:

[0210] Preferably, in formula I-3A,

[0211] R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl;

[0212] X and Y are independently C 3-10 Alkylene;

[0213] Z 1 for * indicates the end connected to X; Z 1a C 1-6 Alkylene;

[0214] Z 2 for * indicates the end connected to Y; Z 2a C 1-6 Alkylene;

[0215] W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene;

[0216] R 2 C 1-15 alkyl or R 2b C 1-15 alkyl;

[0217] R 3 C1-15 alkyl;

[0218] R 4 C 1-15 alkyl or R 4d C 1-15 alkyl;

[0219] R 5 C 1-15 alkyl;

[0220] Preferably, in formula I-3B,

[0221] R 1 It is a hydroxyl group;

[0222] X and Y are independently C 3-10 Alkylene;

[0223] Z 1 for Z 1a C 1-6 Alkylene;

[0224] Z 2 for Z 2a C 1-6 Alkylene;

[0225] W 1 and W 2 It is a chemical bond;

[0226] R 2 C 1-15 alkyl;

[0227] R 3 C 1-15 alkyl;

[0228] R 4 C 1-15 alkyl;

[0229] R 5 C 1-15 alkyl;

[0230] Preferably, in formula I-3C,

[0231] R 1 It is a hydroxyl group;

[0232] X and Y are independently C 3-10 Alkylene;

[0233] Z 1 for * indicates the end connected to X;

[0234] Z2 for * indicates the end connected to Y;

[0235] W 1 and W 2 Independently for C 1-6 Alkylene;

[0236] R 2 for R 2b C 1-15 alkyl;

[0237] R 3 C 1-15 alkyl;

[0238] R 4 for R 4d C 1-15 alkyl;

[0239] R 5 C 1-15 alkyl.

[0240] In one embodiment of the present invention, the compound of formula I is any of the following compounds:

[0241] The present invention also provides the use of the compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a nucleic acid prophylactic agent and / or therapeutic agent delivery carrier;

[0242] The nucleic acid therapeutic and / or preventive agents are 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.

[0243] The present invention also provides a composition comprising substance Z, said substance Z being a compound of formula I as described above or a pharmaceutically acceptable salt thereof.

[0244] The present invention also provides a lipid carrier comprising substance Z, wherein substance Z is a compound of formula I as described above or a pharmaceutically acceptable salt thereof.

[0245] In one embodiment of the present invention, the lipid carrier further includes a diluent. The diluent may be a phosphate buffer or a Tris buffer, etc.

[0246] In one embodiment of the present invention, the lipid carrier further includes phospholipids.

[0247] In one embodiment of the present invention, the phospholipid can be a conventional phospholipid in the art, which is an amphoteric accessory molecule that facilitates the fusion of lipid particles and cell membranes. The phospholipid can be a phospholipid molecule having an electrically polar end and a nonpolar end of a fatty acid chain, such as distearylphosphatidylcholine (DSPC), dimyristoylphosphocholine (DMPC), dioleoylphosphocholine (DOPC), palmitoylphosphocholine (DPPC), 1,2-distearylphosphocholine (DSPC), docosanoylphosphocholine (DUPC), or palmitoylphosphocholine (POPC), etc.

[0248] In one embodiment of the present invention, the lipid carrier further includes PEG lipids (polyethylene glycol modified lipids).

[0249] In one embodiment of the present invention, the PEG lipid may be a lipid molecule modified with a polyethylene glycol hydrophilic end. 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).

[0250] In one embodiment of the present invention, the lipid carrier further includes sterols.

[0251] 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, tomatine, ursolic acid, and α-tocopherol, such as cholesterol.

[0252] In one embodiment of the present invention, the molar ratio of substance Z to sterol in the lipid carrier is 0.5-5:1, preferably 0.5-3:1, for example 1.3:1.

[0253] In one embodiment of the present invention, the molar ratio of substance Z to phospholipid in the lipid carrier is 1-15:1, preferably 2-8:1, for example 3-6:1.

[0254] In one embodiment of the present invention, the molar ratio of substance Z to phospholipid in the lipid carrier is 1-15:1, preferably 1-10:1, for example 5:1.

[0255] In one embodiment of the present invention, the molar ratio of substance Z to PEG lipid in the lipid carrier is 10-100:1, preferably 10-50:1, for example 33.3:1.

[0256] In this invention, molar content refers to the percentage of a substance in the total mass of the lipid carrier, and the sum of the molar contents of all components in the lipid carrier does not exceed 100 mol.

[0257] In one embodiment of the present invention, the molar content of substance Z in the lipid carrier is 30 mol% to 70 mol%, for example, 50 mol%.

[0258] In one embodiment of the present invention, the phospholipid molar content in the lipid carrier is from 5 mol% to 20 mol%, for example, 10 mol%.

[0259] In one embodiment of the invention, the sterol content in the lipid carrier is 20 mol% to 60 mol%, for example, 38.5 mol%.

[0260] In one embodiment of the present invention, the PEG lipid in the lipid carrier has a molar content of about 0.2 mol% to 5 mol%, for example, 1.5 mol%.

[0261] In one embodiment of the present invention, the lipid carrier is composed of the substance Z, the diluent, the phospholipid, the PEG lipid, and the sterol.

[0262] In one embodiment of the present invention, the lipid carrier has the following formulation: Substance Z: Phospholipid: Sterol: PEG lipid is 50:10:38.5:1.5;

[0263] The preferred substance Z is... The phospholipid is preferably DSPC; the sterol is preferably cholesterol; and the PEG lipid is preferably DMG-PEG2000.

[0264] The present invention also provides the use of the lipid carrier in the preparation of nucleic acid prophylactic and / or therapeutic delivery carriers;

[0265] The nucleic acid therapeutic and / or preventive agents are 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.

[0266] The present invention also provides a lipid nanoparticle comprising a nucleic acid preventive agent and / or therapeutic agent and the aforementioned lipid carrier;

[0267] The nucleic acid therapeutic and / or preventive agents are 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.

[0268] In one aspect of this invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles can be (2-30):1. This nitrogen-to-phosphorus ratio refers to the ratio of the molar number of ionizable nitrogen atoms in one or more ionizable lipid compounds to the molar number of phosphate groups in RNA. In this application, it is the ratio of the molar number of ionizable nitrogen atoms in the ionizable lipid nanoparticles to the molar number of phosphate groups in the mRNA within the pharmaceutical composition. Preferably, the nitrogen-to-phosphorus ratio is (2-20):1, more preferably (3-20):1, for example (3-16):1.

[0269] In one embodiment of the present invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles is 6:1.

[0270] 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, more preferably 50-80 nm, for example 71.19 nm, 59.77 nm, 60.70 nm, 62.58 nm or 65.27 nm.

[0271] In one embodiment of the present invention, the lipid nanoparticles have a particle size (average particle size) of 40-150 nm, such as 51.06 nm, 54.04 nm, 56.72 nm, 57.13 nm, 58.09 nm, 61.61 nm, 62.63 nm, 64.82 nm, 66.13 nm, 68.31 nm, 70.58 nm, 71.20 nm, 77.44 nm, 77.93 nm, 80.03 nm, 84.49 nm, 87.13 nm, 87.86 nm, 89.65 nm, 98.29 nm, or 118.03 nm.

[0272] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.001-0.15, for example 0.036, 0.043, 0.068, 0.072 or 0.101.

[0273] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.001-0.30, for example, 0.023, 0.038, 0.041, 0.046, 0.053, 0.053, 0.056, 0.058, 0.060, 0.069, 0.087, 0.088, 0.090, 0.091, 0.092, 0.096, 0.097, 0.103, 0.105, 0.108, 0.122, 0.124, 0.125, 0.130, 0.144, 0.179, 0.206 or 0.224.

[0274] In one aspect of the present invention, the encapsulation efficiency of the lipid nanoparticles is 90%-100%, for example 94.4%, 95.7%, 96.8%, 95.8% or 96.1%.

[0275] In one aspect of the present invention, the encapsulation efficiency of the lipid nanoparticles is 85%-100%, for example, 87.3%, 89.0%, 89.1%, 89.4%, 90.9%, 92.1%, 92.1%, 92.2%, 92.4%, 93.4%, 94.1%, 95.2%, 95.3%, 95.5%, 95.7%, 95.7%, 96.0%, 96.2%, 96.2%, 96.2%, 96.6%, 96.7%, 96.8%, 96.9%, 96.9%, 97.1%, 97.7%, or 97.9%.

[0276] In one embodiment of the present invention, the lipid nanoparticles contain a lipid carrier that encapsulates the nucleic acid preventive agent and / or therapeutic agent.

[0277] Unless otherwise specified, the terms used in this invention may be defined as follows:

[0278] Those skilled in the art will understand that, according to conventions used in the art, the structural formulas of the groups described in this invention are... This refers to the fact that the corresponding group is connected to other fragments or groups in the compound through this site.

[0279] In this article, the substituents used may be preceded by a single dash "-" to indicate that the named substituent is connected to the parent moiety by a single bond.

[0280] When one of the variables is selected as a chemical bond, it means that the two groups connected to it are directly linked.

[0281] When any variable appears multiple times in the definition of a compound, the definition of the variable at each position is independent of the definitions at other positions, and their meanings are independent of each other.

[0282] The term "multiple" refers to 2, 3, or 4.

[0283] The term "alkyl" refers to a straight-chain or branched 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, etc.

[0284] The term "alkylene" refers to a straight-chain divalent hydrocarbon group or a branched divalent hydrocarbon group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, 1-methylpropylene, butylene, etc.

[0285] The term "alkenyl" refers to a straight-chain or branched hydrocarbon group having one or more double bonds with a specific number of carbon atoms (e.g., C2-C4). These carbon-carbon double bonds can be internal or terminal.

[0286] The term "cycloalkylene" refers to a saturated cyclic, divalent group having a specified number of carbon atoms in the ring (e.g., C3-C8) consisting solely of carbon atoms. Examples of cycloalkyl groups include, but are not limited to, those listed below. wait.

[0287] The term "pharmaceutically acceptable salt" refers to a salt prepared from the compounds of the present invention with a relatively non-toxic, pharmaceutically acceptable acid or base. 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 a pure solution or 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 a pure solution or a suitable inert solvent.

[0288] The term “treatment” refers to a therapeutic approach. When a specific condition is involved, treatment means: (1) alleviating one or more biological manifestations of the disease or condition; (2) interfering with (a) one or more points in a biological cascade that causes or precipitates the condition or (b) one or more biological manifestations of the condition; (3) improving 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 development of the condition or one or more biological manifestations of the condition.

[0289] The term "prevention" refers to a period of time during which a subject remains healthy relative to the disease or condition mentioned herein. It should be understood that this period of time depends on the amount of pharmaceutical compound administered and 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 compound according to the invention. However, the term requires, preferably, the effective prevention of a statistically significant portion of a cohort or group of subjects from developing the disease or condition referred to herein or its accompanying symptoms. Preferably, in this case, a group or cluster of subjects is anticipated who would typically, i.e., without taking the preventive measures according to the invention, develop the disease or condition referred to herein. Those skilled in the art can readily determine whether a portion is statistically significant using various well-known statistical evaluation tools discussed elsewhere in this specification.

[0290] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0291] The reagents and raw materials used in this invention are all commercially available.

[0292] The positive and progressive effects of this invention are as follows: the LNP formulation prepared using the nitrogen-containing chain compound of this invention has relatively uniform nanoparticle size, high encapsulation efficiency, and high in vivo expression activity. Attached Figure Description

[0293] Figure 1 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 1, 2, 3, and 5;

[0294] Figure 2 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 4 and 14;

[0295] Figure 3 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 6, 7, 10, 12, 13, 17, 19, 21, and 22.

[0296] Figure 4 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 9, 20, 25, 26, 27, 28 and 30;

[0297] Figure 5 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds I-6-II, 8, 15, 23, and 29;

[0298] Figure 6 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 11, 16, 18, 31, and 32.

[0299] Figure 7 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 53 and 54;

[0300] Figure 8 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 33, 34, 36, 37, 38, 39, 40, 41, 43, 44, 45, and 46.

[0301] Figure 9 shows the chemiluminescence intensity of the liver region in mice after tail vein administration of compounds 35, 47, 48, 50, 51 and 52;

[0302] Figure 10 shows the expression of RSV antigen in mouse serum after tail vein administration of compounds I-6-II, 7, 10, 17, 21, 1, and 5 (6 hours after administration, n=5). Detailed Implementation

[0303] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0304] Preparation Example 1: Preparation of Compound 1

[0305] Step 1: Preparation of 1-1

[0306] Reaction formula:

[0307] Material proportions:

[0308] Operation process:

[0309] Succinic anhydride, 8-pentadecanol, DMAP, triethylamine, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 16 hours. The product had an Rf value of 0.6, and TLC (DCM:MeOH = 10:1) yielded 28 g of product after column chromatography purification.

[0310] Step 2: Preparation of 1-2

[0311] Reaction formula:

[0312] Material proportions:

[0313] Operation process:

[0314] 1,1-,6-bromohexanol, EDCI, DMAP, and DCM were added to the reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.5 and was measured by TLC (PE:EA = 10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After concentration, the product was purified by column chromatography to obtain 2.7 g of oil.

[0315] Step 3: Preparation of Compound 1

[0316] Reaction formula:

[0317] Material proportions:

[0318] Operation process:

[0319] 1-2, cis-3-aminocyclohexanol hydrochloride, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 65 °C for 16 h. The product rR value was 0.6, and the TLC (DCM:MeOH = 20:1) was measured. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.1 g of oil.

[0320] 1 H NMR(400MHz,Chloroform-d)δ4.86(p,J=6.2Hz,2H),4.07(t,J=6.7Hz,4H),3.65(dp,J=8.8,4.1Hz,1H),2.61(s,8H),2. 53(s,3H),1.93–1.68(m,4H),1.63(q,J=7.1Hz,5H),1.49(t,J=6.3Hz,11H),1.41–1.18(m,54H),0.87(t,J=6.7Hz,12H).

[0321] MS(ES+)m / z): 936.7 (M) + .

[0322] Preparation Example 2: Preparation of Compound 2

[0323] Step 1: Preparation of 2-1

[0324] Reaction formula:

[0325] Material proportions:

[0326] Operation process:

[0327] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the mixture was stirred at room temperature for 16 h. The TLC (PE:EA = 4:1) yielded an Rf value of 0.4. The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 27 g of a colorless oil.

[0328] Step 2: Preparation of 2-2

[0329] Reaction formula:

[0330] Material proportions:

[0331] Operation process:

[0332] 2-1, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The Rf value of the product by TLC (PE:EA = 20:1) was 0.6. The reaction solution was washed once with 200 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 20 g of colorless oil.

[0333] Step 3: Preparation of Compound 2

[0334] Reaction formula:

[0335] Material proportions:

[0336] Operation process:

[0337] 2-2, cis-3-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 65 °C for 16 h. The TLC (DCM:MeOH = 10:1) product Rf value was 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the product was purified by column chromatography to obtain 1 g of oil.

[0338] 1H NMR(400MHz,Chloroform-d)δ5.07(dt,J=9.9,4.8Hz,2H),4.21(dd,J=11.9,3.3Hz,2H),4.01(dd,J=11.8,6.8Hz,2H),3.69–3.59( m,1H),2.52(s,4H),2.30(t,J=7.5Hz,8H),2.07–1.75(m,4H),1.67–1.43(m,16H),1.27(d,J=16.2Hz,56H),0.87(t,J=6.6Hz,12H).

[0339] MS(ES+)m / z): 965.8(M+H) + .

[0340] Preparation Example 3: Preparation of Compound 3

[0341] Step 1: Synthesis of 1-(benzyloxy)dodecane-3-ol

[0342] Reaction formula:

[0343] Material proportions:

[0344] Operation process:

[0345] 3-(benzyloxy)propanal (6.00 g, 36.5 mmol) was dissolved in tetrahydrofuran (60.0 mL), and a tetrahydrofuran solution of nonylmagnesium bromide (10.1 g, 43.8 mmol) was slowly added dropwise at 0 °C. The reaction was then carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-(benzyloxy)dodecane-3-ol (5.87 g).

[0346] Step 2: Synthesis of 1-(benzyloxy)dodecane-3-yloctyl ester

[0347] Reaction formula:

[0348] Material proportions:

[0349] Operation process:

[0350] 1-(benzyloxy)dodecane-3-ol (5.87 g, 20.0 mmol) and n-octanoic acid (3.47 g, 24.0 mmol) were dissolved in dichloromethane (58.0 mL), followed by the sequential addition of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.77 g, 30.1 mmol) and 4-dimethylaminopyridine (245 mg, 2.01 mmol). The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid, 1-(benzyloxy)dodecane-3-yloctyl ester (6.00 g).

[0351] Step 3: Synthesis of 1-hydroxydodecane-3-yloctyl ester

[0352] Reaction formula:

[0353] Material proportions:

[0354] Operation process:

[0355] 1-(benzyloxy)dodecane-3-yloctyl ester (6.00 g, 14.3 mmol) was dissolved in methanol (60.0 mL), and palladium / carbon (3.05 g, 2.87 mmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. The solution was then evaporated to dryness to obtain a colorless liquid, 1-hydroxydodecane-3-yloctyl ester (4.60 g).

[0356] Step 4: Synthesis of 1-[(6-bromohexanoyl)oxo]dodecane-3-yloctyl ester

[0357] Reaction formula:

[0358] Material proportions:

[0359] Operation process:

[0360] 1-Hydroxydodecane-3-yloctyl ester (2.00 g, 6.09 mmol) was dissolved in dichloromethane (15.0 mL), followed by the addition of 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). The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid 1-[(6-bromohexanoyl)oxo]dodecane-3-yloctyl ester (2.78 g).

[0361] Step 5: Synthesis of 1-[(6-{[(1R,3S)-3-hydroxycyclohexyl](6-{[3-(octanoyloxo)dodecyl]oxo}-6-oxoylidexyl)amino}hexanoyl)oxo]dodecane-3-yloctyl ester]

[0362] Reaction formula:

[0363] Material proportions:

[0364] Operation process:

[0365] 1-[(6-bromohexanoyl)oxo]dodecane-3-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (10.0 mL), and (1S,3R)-3-aminocyclohexane-1-ol (143 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 reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a colorless liquid 1-[(6-{[(1R,3S)-3-hydroxycyclohexyl](6-{[3-(octanoyloxo)dodecyl]oxo}-6-oxylidenehexyl)amino}hexanoyl)oxo]dodecane-3-yloctyl ester (441 mg, 21.6% yield).

[0366] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.98 (br s, 2H), 4.09 (t, J = 6.8Hz, 4H), 3.68 (br d, J = 3.8Hz, 1H), 2.89 (br s,4H),2.40-2.27(m,8H),2.01-1.81(m,8H),1.72-1.48(m,18H),1.37-1.15(m,52H),0.88(t,J=6.8Hz,12H)ppm.

[0367] LCMS:RT=2.175,m / z 964.8[M+H] + .

[0368] Preparation Example 4: Preparation of Compound 5

[0369] Step 1: Preparation of 5-1

[0370] Reaction formula:

[0371] Material proportions:

[0372] Operation process:

[0373] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0374] Step 2: Preparation of compound 5-2

[0375] Reaction formula:

[0376] Material proportions:

[0377] Operation process:

[0378] 5-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 4 g of oil.

[0379] Step 3: Preparation of Compound 5

[0380] Reaction formula:

[0381] Material proportions:

[0382] Operation process:

[0383] 5-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 65 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 20:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.8 g of oil.

[0384] 1H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.8,3.3Hz,2H),4.21(dd,J=11.8,3.4Hz,2H),4.02(dd,J=11.8,6.8Hz,3 H),2.30(t,J=7.5Hz,8H),1.84(d,J=13.4Hz,2H),1.66–1.45(m,20H),1.35–1.21(m,54H),0.87(t,J=6.7Hz,12H).

[0385] MS(ES+)m / z): 964.8 (M) + .

[0386] Preparation Example 5: Preparation of Compound 4

[0387] Step 1: Synthesis of 2-(hexanoyloxo)dodecyl 6-[(6-{[2-(hexanoyloxo)dodecyl]oxo}-6-oxoylide)[(1R,3S)-3-hydroxycyclohexyl]amino]hexyl ester

[0388] Reaction formula:

[0389] Material proportions:

[0390] Operation process:

[0391] 2-(hexanoyloxy)dodecyl 6-bromohexyl ester (770 mg, 1.61 mmol) was dissolved in acetonitrile (6.0 mL), and (1S,3R)-3-aminocyclohexane-1-ol (110 mg, 0.725 mmol), potassium carbonate (668 mg, 4.84 mmol), potassium iodide (401 mg, 2.42 mmol), and tetrahydrofuran (2.0 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a colorless liquid 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxo}-6-oxylidenehexyl)[(1R,3S)-3-hydroxycyclohexyl]amino]hexyl ester (400 mg, 25.8% yield).

[0392] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.15-5.02 (m, 2H), 4.23 (dd, J = 3.2, 11.6Hz, 2H), 4.03 (dd, J = 6.8, 12.0Hz, 2H), 3.65 (br s,1H),2.58-2.38(m,4H),2.31(t,J=7.6Hz,8H),2.06-1.72(m,4H),1.71-1.59(m,13H),1.50-1.18(m,53H),0.90(q,J=7.2Hz,12H).

[0393] LCMS:RT=3.994,m / z 908.7[M+H] + .

[0394] Preparation Example 6: Preparation of Compound 14

[0395] Step 1: Synthesis of 1-(benzyloxy)-3-(octanoyloxy)propane-2-yloctyl ester

[0396] Reaction formula:

[0397] Material proportions:

[0398] Operation process:

[0399] 2-{[(hexyloxy)carbonyl]oxo}dodecane-1-ol (1.00 g, 5.49 mmol) and octanoic acid (1.74 g, 12.0 mmol) were dissolved in dichloromethane (10 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.16 g, 16.4 mmol) and 4-dimethylaminopyridine (134 mg, 1.10 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-(benzyloxy)-3-(octanoyloxy)propane-2-yloctyl ester (2.30 g, 96.4% yield).

[0400] Step 2: Synthesis of 1-hydroxy-3-(octanoyloxy)propane-2-yloctyl ester

[0401] Reaction formula:

[0402] Material proportions:

[0403] Operation process:

[0404] 1-(benzyloxy)-3-(octanoyloxy)propane-2-yloctyl ester (2.30 g, 5.29 mmol) was dissolved in methanol (25 mL), and palladium on carbon (1.13 g, 1.06 mmol) was added. The reaction was carried out at 35 °C and 40 Psi for 12 h under hydrogen protection. The reaction was monitored by TLC until completion, filtered, and concentrated. The colorless liquid 1-hydroxy-3-(octanoyloxy)propane-2-yloctyl ester (1.80 g, 98.7% yield) was purified by column chromatography.

[0405] Step 3: Synthesis of 1-[(6-bromohexanoyl)oxo]-3-(octanoyloxo)propane-2-yloctyl ester

[0406] Reaction formula:

[0407] Material proportions:

[0408] Operation process:

[0409] 1-Hydroxy-3-(octanoyloxy)propane-2-yloctyl ester (1.80 g, 5.23 mmol) and 6-bromohexanoic acid (1.22 g, 6.27 mmol) were dissolved in dichloromethane (20 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.50 g, 7.84 mmol) and 4-dimethylaminopyridine (127 mg, 1.05 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-[(6-bromohexanoyl)oxy]-3-(octanoyloxy)propane-2-yloctyl ester (2.60 g, 98.0% yield).

[0410] Step 4: Synthesis of 1-{[6-({6-[2,3-di(octanoyloxy)propoxy]-6-oxylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexanoyl]oxo}-3-(octanoyloxy)propane-2-yloctyl ester

[0411] Reaction formula:

[0412] Material proportions:

[0413] Operation process:

[0414] 3-[(6-bromohexanoyl)oxo]-2-(octyloxy)propyl octyl ester (1.00 g, 1.97 mmol) was dissolved in acetonitrile (7 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (102 mg, 0.886 mmol), potassium carbonate (816 mg, 5.91 mmol), potassium iodide (490 mg, 2.96 mmol), and tetrahydrofuran (3 mL). The reaction was carried out at 75 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid 1-{[6-({6-[2,3-di(octyloxy)propoxy]-6-oxylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexanoyl]oxo}-3-(octyloxy)propane-2-yl octyl ester (446 mg, 20.8% yield).

[0415] 1 H NMR(400MHz, CHLOROFORM-d)δ=5.31-5.23(m,2H),4.30(ddd,J1=1.2,J2=4.4,J3=12.0Hz,4H),4.20-4.12(m,4H),4.06-3.98(m ,1H),2.57-2.38(m,3H),2.33(dt,J1=3.6,J2=7.6Hz,12H),1.94-1.80(m,2H),1.75-1.59(m,16H),1.57-1.46(m,8H),1.30(br s,37H),0.98-0.80(m,12H)ppm.

[0416] LCMS:RT=2.411,m / z 997.2[M+H] + .

[0417] Preparation Example 7: Preparation of Compound 6

[0418] Step 1: Preparation of 6-1

[0419] Reaction formula:

[0420] Material proportions:

[0421] Operation process:

[0422] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0423] Step 2: Preparation of 6-2

[0424] Reaction formula:

[0425] Material proportions:

[0426] Operation process:

[0427] 6-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 4 g of oil.

[0428] Step 3: Preparation of Compound 6

[0429] Reaction formula:

[0430] Material proportions:

[0431] Operation process:

[0432] 6-2,2-aminocyclohexanol, K₂CO₃, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 75°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.2 g of a colorless oil.

[0433] 1H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.6,3.3Hz,2H),4.22(dt,J=11.9,2.9Hz,2H),4.03(ddd,J=11.8,6.8,2.2Hz,2H),3.29(dq,J=9.8,4.5Hz,1H), 2.48(ddd,J=12.9,9.0,6.8Hz,2H),2.30(dd,J=8.4,6.7Hz,10H),2.11(d,J= 7.1Hz,1H),1.78–1.54(m,16H),1.42–1.20(m,58H),0.88(t,J=6.6Hz,12H).

[0434] MS(ES+)m / z): 964.0 (M) + .

[0435] Preparation Example 8: Preparation of Compound 7

[0436] Step 1: Preparation of 7-1

[0437] Reaction formula:

[0438] Material proportions:

[0439] Operation process:

[0440] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0441] Step 2: Preparation of 7-2

[0442] Reaction formula:

[0443] Material proportions:

[0444] Operation process:

[0445] 7-1, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 4 g of oil.

[0446] Step 3: Preparation of Compound 7

[0447] Reaction formula:

[0448] Material proportions:

[0449] Operation process:

[0450] 7-2,3-aminocyclopentanol, K₂CO₃, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 75°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 800 mg of a colorless oil.

[0451] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.8,3.3Hz,2H),4.39(s,1H),4.22(dd,J=11.8,3.4Hz,2H),4.01(dd,J=11.8,6.8Hz,2 H),2.57(s,3H),2.30(td,J=7.5,2.8Hz,8H),1.90–1.70(m,4H),1.67–1.45(m,18H),1.35–1.21(m,54H),0.88(t,J=6.6Hz,12H).

[0452] MS(ES+)m / z): 950.0(M) + .

[0453] Preparation Example 9: Preparation of Compound 10

[0454] Step 1: Preparation of 10-1

[0455] Reaction formula:

[0456] Material proportions:

[0457] Operation process:

[0458] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0459] Step 2: Preparation of 10⁻²

[0460] Reaction formula:

[0461] Material proportions:

[0462] Operation process:

[0463] 10⁻¹, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 4 g of oil.

[0464] Step 3: Preparation of compound LQ210-54

[0465] Reaction formula:

[0466] Material proportions:

[0467] Operation process:

[0468] 10⁻², cis-4-aminocyclohexane methanol hydrochloride, K₂CO₃, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 75 °C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 800 mg of a colorless oil.

[0469] 1H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.3Hz,2H),4.22(dd,J=11.8,3.3Hz,2H),4.02(dd,J=11.8,6.8Hz,2H),3.62(d,J=6.7Hz, 2H),2.66–2.53(m,3H),2.30(td,J=7.5,2.8Hz,8H),1.85–1.76(m,4H),1.66–1.50(m,21H),1.34–1.22(m,54H),0.88(t,J=6.6Hz,12H).

[0470] MS(ES+)m / z): 978.0(M) + .

[0471] Preparation Example 10: Preparation of Compound 12

[0472] Step 1: Preparation of 12-1

[0473] Reaction formula:

[0474] Material proportions:

[0475] Operation process:

[0476] Heptadecanol, 1,6-adipic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the reaction was carried out at room temperature for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.6. The reaction solution was washed once with 200 mL of aqueous solution, and the organic phase was dried over anhydrous magnesium sulfate. After filtration, concentration, and purification by column chromatography, 7 g of oil was obtained.

[0477] Step 2: Preparation of 12-2

[0478] Reaction formula:

[0479] Material proportions:

[0480] Operation process:

[0481] 12-1, 4-bromobutanol, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 3.5 g of oil.

[0482] Step 3: Preparation of 12-3

[0483] Reaction formula:

[0484] Material proportions:

[0485] Operation process:

[0486] 1-Undecyl alcohol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7.5 g of an oily substance.

[0487] Step 4: Preparation of 12-4

[0488] Reaction formula:

[0489] Material proportions:

[0490] Operation process:

[0491] 12-3, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 3.5 g of a colorless oil.

[0492] Step 5: Preparation of Compound 12

[0493] Reaction formula:

[0494] Material proportions:

[0495] Operation process:

[0496] 12⁻², 12⁻⁴, K₂CO₃, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded a product Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.2 g of a colorless oil.

[0497] 1H NMR(400MHz,Chloroform-d)δ4.85(p,J=6.3Hz,1H),4.06(dt,J=9.3,6.7Hz,4H),4.00(s,1H),2.48(s,3H),2.3 1(qd,J=7.2,4.1Hz,6H),1.85(d,J=13.3Hz,2H),1.69–1.46(m,24H),1.36–1.20(m,44H),0.87(t,J=6.6Hz,9H).

[0498] MS(ES+)m / z): 822.0(M) + .

[0499] Preparation Example 11: Preparation of Compound 13

[0500] Step 1: Synthesis of ({[1,3-di(heptoxy)propane-2-yl]oxo}methyl)benzene

[0501] Reaction formula:

[0502] Material proportions:

[0503] Operation process:

[0504] 2-(benzyloxy)propane-1,3-diol (2.00 g, 10.9 mmol) was dissolved in N,N-dimethylformamide (20 mL), and sodium hydroxide (1.32 g, 32.9 mmol) was added at 25 °C. The reaction was continued at 25 °C for 1 hour. 7-bromoheptane-1-ol (4.32 g, 24.1 mmol) was dissolved in N,N-dimethylformamide (10 mL) and added to the reaction mixture. Finally, the reaction was carried out under nitrogen protection at 25 °C for 12 hours. TLC monitoring showed the formation of new spots. 300 mL of water was added, and the mixture was extracted twice with 100 mL of ethyl acetate each time. The organic phase was concentrated. The crude product was subjected to column chromatography to obtain a yellow liquid ({[1,3-di(heptyloxy)propane-2-yl]oxo}methyl)benzene (2.30 g, 55.3% yield).

[0505] Step 2: Synthesis of 1,3-di(heptoxy)propane-2-ol

[0506] Reaction formula:

[0507] Material proportions:

[0508] Operation process:

[0509] 1-(benzyloxy)-3-(octanoyloxy)propane-2-yloctyl ester (2.30 g, 5.29 mmol) was dissolved in methanol (25 mL), and palladium on carbon (1.13 g, 1.06 mmol) was added. The mixture was reacted at 35 °C and 40 Psi for 12 hours under hydrogen protection. The reaction was monitored by TLC until completion, and then filtered and concentrated. The solution was purified by column chromatography to obtain a colorless liquid 1,3-di(heptoxy)propane-2-ol (1.60 g, 91.3% yield).

[0510] Step 3: Synthesis of 1,3-di(heptoxy)propane-2-yl 8-bromooctyl ester

[0511] Reaction formula:

[0512] Material proportions:

[0513] Operation process:

[0514] 1,3-Di(heptoxy)propane-2-ol (1.60 g, 5.55 mmol) and 8-bromooctanoic acid (1.48 g, 6.66 mmol) were dissolved in dichloromethane (20 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.59 g, 8.32 mmol) and 4-dimethylaminopyridine (135 mg, 1.11 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1,3-di(heptoxy)propane-2-yl 8-bromooctyl ester (2.60 g, 98.0% yield).

[0515] Step 4: Synthesis of undecyl 6-bromohexyl ester

[0516] Reaction formula:

[0517] Material proportions:

[0518] Operation process:

[0519] Undecane-1-ol (1.77 g, 10.2 mmol) and 6-bromohexanoic acid (2.00 g, 10.2 mmol) were dissolved in dichloromethane (20 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.95 g, 15.3 mmol) and 4-dimethylaminopyridine (250 mg, 2.05 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give undecyl 6-bromohexyl ester (3.37 g, 94.0% yield) as a colorless liquid.

[0520] Step 5: Synthesis of undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester

[0521] Reaction formula:

[0522] Material proportions:

[0523] Operation process:

[0524] Undecyl 6-bromohexyl ester (1.30 g, 3.72 mmol) and (1s,4s)-4-aminocyclohexane-1-ol (857 mg, 7.44 mmol) were dissolved in acetonitrile (13 mL). Triethylamine (753 mg, 7.44 mmol) was added to the reaction solution. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the organic phase was concentrated. The crude product was purified by column chromatography to obtain a yellow liquid undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (460 mg, 29.6% yield).

[0525] Step Six: Synthesis of 1,3-Di(heptoxy)propane-2-yl-8-{[6-oxoylide-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester

[0526] Reaction formula:

[0527] Material proportions:

[0528] Operation process:

[0529] Undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (400 mg, 1.04 mmol) was dissolved in acetonitrile (4 mL), and 1,3-di(heptoxy)propane-2-yl 8-bromooctyl ester (617 mg, 1.25 mmol), potassium carbonate (504 mg, 3.65 mmol), potassium iodide (207 mg, 1.25 mmol), and tetrahydrofuran (2 mL) were added sequentially. The reaction was carried out at 75 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a colorless liquid 1,3-di(heptoxy)propane-2-yl 8-{[6-oxoylide-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester (57 mg, 6.30% yield).

[0530] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.13 (quin, J = 5.2Hz, 1H), 4.10-3.99 (m, 3H), 3.60-3.52 (m, 4H), 3.51-3.36 (m, 4H), 2.65-2.39 (m, 3H), 2.32(td,J1=7.6,J2=10.0Hz,4H),1.96-1.82(m,3H),1.70-1.61(m,9H),1.58-1.49(m,9H),1.39-1.23(m,43H),0.92-0.86(m,9H)ppm.

[0531] LCMS:RT=2.628,m / z 796.6[M+H] + .

[0532] Preparation Example 12: Preparation of Compound 17

[0533] Step 1: Preparation of 17-1

[0534] Reaction formula:

[0535] Material proportions:

[0536] Operation process:

[0537] Succinic anhydride, 8-pentadecanol, DMAP, triethylamine, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 6.5 g of a colorless oil.

[0538] Step 2: Preparation of 17-2

[0539] Reaction formula:

[0540] Material proportions:

[0541] Operation process:

[0542] 17-1, 6-bromohexanol, EDCI, DMAP, and DCM were added to the reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.5 and was measured by TLC (PE:EA = 10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 5.6 g of a colorless oil.

[0543] Step 3: Preparation of Compound 17

[0544] Reaction formula:

[0545] Material proportions:

[0546] Operation process:

[0547] 17-2, cis-4-aminocyclohexanol, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 75°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 900 mg of a colorless oil.

[0548] 1 H NMR(400MHz,Chloroform-d)δ4.89(p,J=6.3Hz,2H),4.09(t,J=6.7Hz,4H),4.02(t,J=3.1Hz,1H),2.63( s,8H),2.52(s,3H),1.92–1.81(m,2H),1.75–1.45(m,22H),1.41–1.21(m,50H),0.89(t,J=6.8Hz,12H).

[0549] MS(ES+)m / z): 936.0(M) + .

[0550] Preparation Example 13: Preparation of Compound 19

[0551] Step 1: Synthesis of hexyl-4-nitrobenzene carbonate

[0552] Reaction formula:

[0553] Material proportions:

[0554] Operation process:

[0555] Hexane-1-ol (2.00 g, 19.6 mmol) was dissolved in dichloromethane (10 mL), and 4-methylbenzenesulfonyl chloride (4.78 g, 39.1 mmol) and chlorinated 4-nitrophenyl ester (4.73 g, 23.5 mmol) were added at 0 °C. The reaction was then carried out at 25 °C for 2 hours under nitrogen protection, and the reaction was monitored by TLC until completion. The mixture was washed with water, extracted, and concentrated. The crude product, hexyl-4-nitrobenzene carbonate (5.00 g, 95.6% yield), was used directly in the next step.

[0556] Step 2: Synthesis of 1-(benzyloxy)dodecane-2-ylhexyl carbonate

[0557] Reaction formula:

[0558] Material proportions:

[0559] Operation process:

[0560] 1-(benzyloxy)dodecane-2-ol (6.84 g, 18.7 mmol) was dissolved in dichloromethane (10 mL), and hexyl 4-nitrobenzene carbonate (5.00 g, 18.7 mmol) was slowly added at 0 °C. The reaction was then carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain 1-(benzyloxy)dodecane-2-ylhexyl carbonate (3.50 g, 44.5% yield).

[0561] Step 3: Synthesis of 2-{[(hexyloxy)carbonyl]oxo}dodecane-1-ol

[0562] Reaction formula:

[0563] Material proportions:

[0564] Operation process:

[0565] 1-(benzyloxy)dodecane-2-ylhexyl carbonate (3.50 g, 8.30 mmol) was dissolved in ethanol (25 mL), and palladium on carbon (966 mg, 1.66 μmmol) was added. The reaction was carried out at 25 °C and 35 Psi for 12 h under hydrogen protection. The reaction was monitored by TLC until completion, filtered, and concentrated. 2-{[(hexyloxy)carbonyl]oxo}dodecane-1-ol (1.50 g, 100% yield) was purified by column chromatography.

[0566] Step 4: Synthesis of 2-{[(hexyloxy)carbonyl]oxo}dodecyl 6-bromohexyl ester

[0567] Reaction formula:

[0568] Material proportions:

[0569] Operation process:

[0570] 2-{[(hexyloxy)carbonyl]oxo}dodecane-1-ol (1.50 g, 4.54 mmol) was dissolved in dichloromethane (20.0 mL), followed by the addition of 6-bromohexanoic acid (1.33 g, 6.81 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.31 g, 6.81 mmol), and 4-dimethylaminopyridine (55.5 mg, 454 μmol). The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain 2-{[(hexyloxy)carbonyl]oxo}dodecyl 6-bromohexyl ester (1.5 g, 58.6% yield).

[0571] Step 5: Synthesis of 2-{[(hexyloxy)carbonyl]oxo}dodecyl 6-({6-[(2-{[(hexyloxy)carbonyl]oxo}dodecyl)oxo]-6-oxoylidexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[0572] Reaction formula:

[0573] Material proportions:

[0574] Operation process:

[0575] 2-{[(hexyloxy)carbonyl]oxo}dodecyl 6-bromohexyl ester (260 mg, 512 μmol) was dissolved in acetonitrile (1 mL), followed by the addition of ethanolamine (26.6 mg, 231 μmol), potassium carbonate (212 mg, 1.54 mmol), potassium iodide (102 mg, 614 μmol), and tetrahydrofuran (3 mL). The reaction was carried out at 75 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The product was purified by column chromatography to obtain 2-{[(hexyloxy)carbonyl]oxo}dodecyl 6-({6-[(2-{[(hexyloxy)carbonyl]oxo}dodecyl)oxo]-6-oxylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (30 mg, 5.93% yield).

[0576] 1 H NMR(400MHz,CHLOROFORM-d)δppm 4.91-4.89(m,1H),4.29-4.26(dd,2H),4.16–4.12(m,4H),4.08–4.05(m,3H),4.48-4.47(d,1H),2. 35–2.33(m,2H),1.68-1.65(m,2H),1.68-1.61(m,25H),1.35–1.26(m,50H),0.90-0.87(m,12H)ppm.

[0577] LCMS: RT=2.510, m / z=969.3[M+H] + .

[0578] Preparation Example 14: Preparation of Compound 21

[0579] Step 1: Synthesis of heptadecan-9-yl-8-[(5-{[4-(heptanoyloxo)dodecyl]oxo}-5-oxoylidenepentyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[0580] Reaction formula:

[0581] Material proportions:

[0582] Operation process:

[0583] 1-[(5-bromopentanoyl)oxo]dodecane-4-ylheptyl ester (1.16 g, 2.42 mmol) was dissolved in acetonitrile (7 mL), and then heptadecan-9-yl 8-{[(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester (1.00 g, 2.02 mmol), potassium carbonate (975 mg, 7.06 mmol), potassium iodide (401 mg, 2.42 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 75 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and then filtered and concentrated. Purification by column chromatography yielded a colorless liquid heptadecan-9-yl 8-[(5-{[4-(heptanoyloxo)dodecyl]oxo}-5-oxoylidenepentyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (416 mg, 22.4% yield).

[0584] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.89 (quind, J1 = 6.4, J2 = 13.2Hz, 2H), 4.10-4.03 (m, 2H), 4.00 (br s, 1H), 2.45 (br s, 4H), 2.35-2.25 (m, 6H), 1.85 (br d,J=12.0Hz,2H),1.69-1.60(m,13H),1.57-1.49(m,9H),1.43(br d,J=3.8Hz,4H),1.37-1.18(m,50H),0.95-0.81(m,12H)ppm.

[0585] LCMS:RT=2.746,m / z 893.4[M+H] + .

[0586] Preparation Example 15: Preparation of Compound 22

[0587] Step 1: Preparation of 22-1

[0588] Reaction formula:

[0589] Material proportions:

[0590] Operation process:

[0591] 7-Bromoheptanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 6 g of oil.

[0592] Step 2: Preparation of 22-2

[0593] Reaction formula:

[0594] Material proportions:

[0595] Operation process:

[0596] 22-1, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 3.8 g of oil.

[0597] Step 3: Preparation of 22-3

[0598] Reaction formula:

[0599] Material proportions:

[0600] Operation process:

[0601] 1-Nonanol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 6.8 g of oil.

[0602] Step 4: Preparation of 22-4

[0603] Reaction formula:

[0604] Material proportions:

[0605] Operation process:

[0606] 22-3, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.6 g of a colorless oil.

[0607] Step 5: Preparation of Compound 22

[0608] Reaction formula:

[0609] Material proportions:

[0610] Operation process:

[0611] 22-4, 22-2, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded a product Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.6 g of a colorless oil.

[0612] 1H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.3Hz,1H),4.21(dd,J=11.8,3.4Hz,1H),4.08–3.98(m,4H),2.45(d,J= 7.5Hz,3H),2.30(t,J=7.5Hz,6H),1.86(d,J=13.4Hz,2H),1.77–1.44(m,22H),1.39–1.19(m,40H),0.91–0.84(m,9H).

[0613] MS(ES+)m / z): 766.0 (M) + .

[0614] Preparation Example 16: Preparation of Compound 9

[0615] Step 1: Preparation of 9-1

[0616] Reaction formula:

[0617] Material proportions:

[0618] Operation process:

[0619] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0620] Step 2: Preparation of 9-2

[0621] Reaction formula:

[0622] Material proportions:

[0623] Operation process:

[0624] 9-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 4 g of oil.

[0625] Step 3: Preparation of Compound 9

[0626] Reaction formula:

[0627] Material proportions:

[0628] Operation process:

[0629] 9-2, cis-4-(aminomethyl)cyclohexanol hydrochloride, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 75°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.3 g of a colorless oil.

[0630] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.3Hz,2H),4.21(dd,J=11.8,3.4Hz,2H),4.02(dd,J=11.8,6.8Hz,2H ),3.96(s,1H),2.30(t,J=7.5Hz,8H),2.20(s,1H),1.67–1.51(m,20H),1.48–1.17(m,62H),0.88(t,J=6.7Hz,12H).

[0631] MS(ES+)m / z): 977.9 (M) + .

[0632] Preparation Example 17: Preparation of Compound 20

[0633] Step 1: Synthesis of ethyl(4Z)-oct-4-en ester

[0634] Reaction formula:

[0635] Material proportions:

[0636] Operation process:

[0637] (4-ethoxy-4-oxoylidenebutyl)triphenylphosphine bromide cation (7.00 g, 18.5 mmol) was dissolved in tetrahydrofuran (70.0 mL), and sodium di(trimethylsilyl)amino (1 M, 19.4 mL) was added at -5 °C. The reaction was carried out at -5 °C for 10 mins. Then, n-butyraldehyde (1.40 g, 19.4 mmol) was added to the reaction solution at -78 °C. The reaction was carried out at 25 °C for 12 h under nitrogen protection. The reaction was monitored by TLC until it ended. The reaction was quenched, extracted twice with ethyl acetate, and the organic phase was concentrated. The solution was purified by column chromatography to obtain colorless liquid ethyl(4Z)-oct-4-en ester (450 mg, 14.2% yield).

[0638] Step 2: Synthesis of (4Z)-oct-4-enoic acid

[0639] Reaction formula:

[0640] Material proportions:

[0641] Operation process:

[0642] Ethyl (4Z)-oct-4-enyl ester (500 mg, 2.94 mmol) was dissolved in tetrahydrofuran (5.00 mL), methanol (0.50 mL), and water (0.50 mL). Lithium hydroxide monohydrate (246 mg, 5.87 mmol) was added, and the mixture was reacted at 60 °C for 12 hours under a nitrogen atmosphere. The reaction was monitored by TLC until it ended. The mixture was then subjected to acid-base back-extraction and concentrated to give a colorless liquid (4Z)-oct-4-enic acid (287 mg, 68.7% yield).

[0643] Step 3: Synthesis of 1-[(tert-butyldimethylsilyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester

[0644] Reaction formula:

[0645] Material proportions:

[0646] Operation process:

[0647] (2R)-1-[(tert-butyldimethylsilyl)oxo]dodecane-2-ol (639 mg, 2.02 mmol) and (4Z)-oct-4-enoic acid (287 mg, 2.02 mmol) were dissolved in dichloromethane (10.0 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (580 mg, 3.03 mmol) and 4-dimethylaminopyridine (24.6 mg, 0.201 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the organic phase was concentrated. Purification by column chromatography yielded a colorless liquid, 1-[(tert-butyldimethylsilyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester (537 mg, 60.3% yield).

[0648] Step 4: Synthesis of 1-hydroxydodecane-2-yl(4Z)-oct-4-en ester

[0649] Reaction formula:

[0650] Material proportions:

[0651] Operation process:

[0652] 1-[(tert-butyldimethylsilyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester (480 mg, 1.09 mmol) was dissolved in tetrahydrofuran (5.00 mL), and triethylamine hydrofluoride (702 mg, 4.36 mmol) was added at 0 °C. The reaction was then carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots. After adjusting the pH to neutral, the mixture was extracted with ethyl acetate, and the organic phase was concentrated. The solution was purified by column chromatography to obtain a colorless liquid, 1-hydroxydodecane-2-yl(4Z)-oct-4-en ester (350 mg, 98.4% yield).

[0653] Step 5: Synthesis of 1-[(8-bromooctanoyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester

[0654] Reaction formula:

[0655] Material proportions:

[0656] Operation process:

[0657] 1-Hydroxydodecane-2-yl(4Z)-oct-4-en ester (350 mg, 1.07 mmol) and 8-bromooctanoic acid (334 mg, 1.50 mmol) were dissolved in dichloromethane (5.00 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (308 mg, 1.61 mmol) and 4-dimethylaminopyridine (13.1 mg, 0.107 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the organic phase was concentrated. The solution was purified by column chromatography to obtain a colorless liquid, 1-[(8-bromooctanoyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester (444 mg, 77.9% yield).

[0658] Step Six: Synthesis of 1-[(8-{[8-({2-[(4Z)-octyl-4-enoxy]dodecyl}oxo)-8-oxoylidecyl][(1s,4s)-4-hydroxycyclohexyl]amino}octanoyl)oxo]dodecane-2-yl(4Z)-octyl-4-en ester]

[0659] Reaction formula:

[0660] Material proportions:

[0661] Operation process:

[0662] 1-[(8-bromooctanoyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester (394 mg, 741 μmol) was dissolved in acetonitrile (4.00 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (40.9 mg, 355 μmol), potassium carbonate (358 mg, 2.59 mmol), potassium iodide (147 mg, 889 μmol), and tetrahydrofuran (2.00 mL). The reaction was carried out at 75 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, then filtered and concentrated. Purified by column chromatography, the colorless liquid 1-[(8-{[8-({2-[(4Z)-oct-4-enyloxy]dodecyl}oxo)-8-oxylideneoctyl][(1s,4s)-4-hydroxycyclohexyl]amino}octanoyl)oxo]dodecane-2-yl(4Z)-oct-4-en ester (202 mg, 23.4% yield) was obtained.

[0663] 1H NMR (400MHz, CHLOROFORM-d) δ=5.48-5.27(m,4H),5.09(br dd,J1=3.4,J2=6.4Hz,2H),4.23(dd,J1=3.4,J2=11.8Hz,2H),4.11(br s,1H),4.03(dd,J1=6.8,J2=11.8Hz,2H),3.29(br d,J=1.0Hz,1H),3.14-2.97(m,2H),2.95-2.81(m,2H),2.43-2.25(m,12H),2.1 2-1.85(m,14H),1.69-1.60(m,6H),1.42-1.20(m,52H),0.95-0.82(m,12H)ppm.

[0664] LCMS:RT=2.751,m / z 1017.9[M+H] + .

[0665] Preparation Example 18: Preparation of Compound 25

[0666] Step 1: Preparation of 25-1

[0667] Reaction formula:

[0668] Material proportions:

[0669] Operation process:

[0670] Heptadecanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and then 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 obtain 9.5 g of an oily substance.

[0671] Step 2: Preparation of 25-2

[0672] Reaction formula:

[0673] Material proportions:

[0674] Operation process:

[0675] 25-1, trans-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.4 g of a colorless oil.

[0676] Step 3: Preparation of 25-3

[0677] Reaction formula:

[0678] Material proportions:

[0679] Operation process:

[0680] 1-Nonanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7 g of oil.

[0681] Step 4: Preparation of Compound 25

[0682] Reaction formula:

[0683] Material proportions:

[0684] Operation process:

[0685] 25-2, 25-3, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded a product Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1 g of a colorless oil.

[0686] 1H NMR(400MHz,Chloroform-d)δ4.85(p,J=6.3Hz,1H),4.05(t,J=6.7Hz,2H),3.56(tt,J=9.2,4.2Hz,1H),2.67–2.37(m,4H),2.27(td,J=7.5 ,5.0Hz,4H),2.08–1.98(m,2H),1.86(s,2H),1.61(q,J=7.1Hz,6H),1.47(h,J=10.1,8.0Hz,8H),1.35–1.21(m,52H),0.87(t,J=6.7Hz,9H).

[0687] MS(ES+)m / z): 764.0 (M) + .

[0688] Preparation Example 19: Preparation of Compound 26

[0689] Step 1: Preparation of 26-1

[0690] Reaction formula:

[0691] Material proportions:

[0692] Operation process:

[0693] Heptadecanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and then 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 obtain 9.5 g of an oily substance.

[0694] Step 2: Preparation of 26-2

[0695] Reaction formula:

[0696] Material proportions:

[0697] Operation process:

[0698] 26-1, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.4 g of a colorless oil.

[0699] Step 3: Preparation of 26-3

[0700] Reaction formula:

[0701] Material proportions:

[0702] Operation process:

[0703] 1-Nonanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7 g of oil.

[0704] Step 4: Preparation of Compound 26

[0705] Reaction formula:

[0706] Material proportions:

[0707] Operation process:

[0708] 26-2, 26-3, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.2 g of a colorless oil.

[0709] 1 H NMR(400MHz,Chloroform-d)δ4.86(p,J=6.3Hz,1H),4.05(t,J=6.7Hz,2H),3.99(t,J=3.1Hz,1H),2.46(s,4H), 2.28(td,J=7.5,5.0Hz,4H),1.84(d,J=13.3Hz,2H),1.67–1.38(m,20H),1.34–1.22(m,48H),0.91–0.84(m,9H).

[0710] MS(ES+)m / z): 764.0 (M) + .

[0711] Preparation Example 20: Preparation of Compound 27

[0712] Step 1: Preparation of 27-1

[0713] Reaction formula:

[0714] Material proportions:

[0715] Operation process:

[0716] Heptadecanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and then 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 obtain 9.5 g of an oily substance.

[0717] Step 2: Preparation of 27-2

[0718] Reaction formula:

[0719] Material proportions:

[0720] Operation process:

[0721] 27-1, trans-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.4 g of a colorless oil.

[0722] Step 3: Preparation of 27-3

[0723] Reaction formula:

[0724] Material proportions:

[0725] Operation process:

[0726] 1-Undecyl alcohol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7.5 g of an oily substance.

[0727] Step 4: Preparation of Compound 27

[0728] Reaction formula:

[0729] Material proportions:

[0730] Operation process:

[0731] 27-2, 27-3, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.2 g of a colorless oil.

[0732] 1 H NMR(400MHz,Chloroform-d)δ4.85(p,J=6.3Hz,1H),4.04(t,J=6.8Hz,2H),3.55(dp,J=9.0,4.2Hz,1H),2.51(d,J=41.9Hz,4H),2.28(q,J=7. 9Hz,4H),2.06–1.97(m,2H),1.83(s,2H),1.61(p,J=7.8,7.1Hz,6H),1.45(dt,J=25.7,5.8Hz,8H),1.35–1.20(m,52H),0.87(t,J=6.8Hz,9H).

[0733] MS(ES+)m / z): 764.0 (M) + .

[0734] Preparation Example 21: Preparation of Compound 28

[0735] Step 1: Preparation of 28-1

[0736] Reaction formula:

[0737] Material proportions:

[0738] Operation process:

[0739] Heptadecanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and then 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 obtain 9.5 g of an oily substance.

[0740] Step 2: Preparation of 28-2

[0741] Reaction formula:

[0742] Material proportions:

[0743] Operation process:

[0744] 28-1, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.4 g of a colorless oil.

[0745] Step 3: Preparation of 28-3

[0746] Reaction formula:

[0747] Material proportions:

[0748] Operation process:

[0749] 1-Undecyl alcohol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7.5 g of an oily substance.

[0750] Step 4: Preparation of Compound 28

[0751] Reaction formula:

[0752] Material proportions:

[0753] Operation process:

[0754] 28-2, 28-3, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded a product Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.3 g of a colorless oil.

[0755] 1H NMR(400MHz,Chloroform-d)δ4.85(p,J=6.3Hz,1H),4.04(t,J=6.7Hz,2H),3.98(s,1H),2.46(t,J=8.2Hz,4H),2 .28(q,J=7.5Hz,4H),1.83(d,J=13.3Hz,2H),1.68–1.38(m,21H),1.27(d,J=18.6Hz,48H),0.87(t,J=6.6Hz,9H).

[0756] MS(ES+)m / z): 764.1 (M) + .

[0757] Preparation Example 22: Preparation of Compound 30

[0758] Step 1: Synthesis of 1-({6-[(6-{[2-(octanoyloxo)dodecyl]oxo}-6-oxoylide)[(1r,4r)-4-hydroxycyclohexyl]amino]hexanoyl}oxo)dodecane-2-yloctyl ester)

[0759] Reaction formula:

[0760] Material proportions:

[0761] Operation process:

[0762] 1-[(6-bromohexanoyl)oxo]dodecane-2-yloctyl ester (2.00 g, 3.96 mmol) was dissolved in acetonitrile (15 mL), and (1r,4r)-4-aminocyclohexane-1-ol (205 mg, 1.78 mmol), potassium carbonate (1.64 g, 11.8 mmol), potassium iodide (985 mg, 5.93 mmol), and tetrahydrofuran (5 mL) were added sequentially. The reaction was carried out at 80 °C for 20 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid 1-({6-[(6-{[2-(octanoyloxo)dodecyl]oxo}-6-oxylidenehexyl)[(1r,4r)-4-hydroxycyclohexyl]amino]hexanoyl}oxo)dodecane-2-yloctyl ester (1.00 g, 25.9% yield).

[0763] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.2, J2 = 6.8Hz, 2H), 4.23 (dd, J1 = 3.2, J2 = 12.0Hz, 2H), 4.08-3.98 (m, 2H), 3.59 (td, J1 = 2.0, J2 = 11.2Hz, 1H),2.54-2.35(m,3H),2.35-2.24(m,9H),2.10-1.98(m,2H),1.87-1.70( m,2H),1.69-1.57(m,12H),1.49-1.18(m,60H),0.89(t,J=6.8Hz,12H)ppm.

[0764] LCMS:RT=2.172,m / z 965.4[M+H] + .

[0765] Preparation Example 23: Preparation of Compound 8

[0766] Step 1: Preparation of 8-1

[0767] Reaction formula:

[0768] Material proportions:

[0769] Operation process:

[0770] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0771] Step 2: Preparation of 8-2

[0772] Reaction formula:

[0773] Material proportions:

[0774] Operation process:

[0775] 8-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 4 g of oil.

[0776] Step 3: Preparation of Compound 8

[0777] Reaction formula:

[0778] Material proportions:

[0779] Operation process:

[0780] 8-2,3-aminocyclobutanol, K₂CO₃, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 75°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1 g of a colorless oil.

[0781] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.9,3.2Hz,2H),4.23(dd,J=11.8,3.3Hz,2H),4.00(dt,J=11.9,5.2Hz,3H),2.94(t,J=7.8Hz ,1H),2.70(dt,J=13.4,8.4Hz,6H),2.47(s,2H),2.30(q,J=6.9Hz,8H),1.68–1.51(m,16H),1.36–1.23(m,52H),0.87(t,J=6.6Hz,12H).

[0782] MS(ES+)m / z): 935.9 (M) + .

[0783] Preparation Example 24: Preparation of Compound 15

[0784] Step 1: Preparation of 15-1

[0785] Reaction formula:

[0786] Material proportions:

[0787] Operation process:

[0788] p-Nitrophenyl chloroformate, 7-bromoheptanol, and DCM were added to a reaction flask under nitrogen protection. Pyridine was added dropwise at room temperature over approximately 5 minutes. After the addition was complete, the mixture was stirred at room temperature for 16 hours. The Rf value of the product was 0.5 by TLC (PE:EA = 6:1). The reaction solution was concentrated and purified by column chromatography to obtain 4 g of an oily substance.

[0789] Step 2: Preparation of 15-2

[0790] Reaction formula:

[0791] Material proportions:

[0792] Operation process:

[0793] 15-1, heptadecano-9-ol, pyridine, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 16 h. The product had an Rf value of 0.6 and was measured by TLC (PE:EA = 10:1). After concentration and purification by column chromatography, 3.5 g of an oily substance was obtained.

[0794] Step 3: Preparation of 15-3

[0795] Reaction formula:

[0796] Material proportions:

[0797] Operation process:

[0798] 1-Undecyl alcohol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to a reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 100 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7.5 g of an oily substance.

[0799] Step 4: Preparation of 15-4

[0800] Reaction formula:

[0801] Material proportions:

[0802] Operation process:

[0803] 15-3, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.4. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 3.5 g of a colorless oil.

[0804] Step 5: Preparation of Compound 15

[0805] Reaction formula:

[0806] Material proportions:

[0807] Operation process:

[0808] 15-2, 15-4, K2CO3, KI, and acetonitrile were added to a reaction flask, and the mixture was heated to 85°C and stirred for 16 h. The TLC (DCM:MeOH = 10:1) yielded an Rf value of 0.5. The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.2 g of a colorless oil.

[0809] 1 H NMR(400MHz,Chloroform-d)δ4.67(p,J=6.2Hz,1H),4.10(t,J=6.7Hz,2H),4.03(q,J=8.3,7.5Hz,3H),2.52(s,4H) ,2.29(t,J=7.5Hz,2H),1.86(d,J=13.5Hz,2H),1.72–1.45(m,18H),1.28(d,J=20.5Hz,50H),0.87(t,J=6.6Hz,9H).

[0810] MS(ES+)m / z): 780.0(M) + .

[0811] Preparation Example 25: Preparation of Compound 23

[0812] Step 1: Synthesis of dioctyl propylene glycol ester

[0813] Reaction formula:

[0814] Material proportions:

[0815] Operation process:

[0816] Octane-1-ol (5.01 g, 38.4 mmol) was dissolved in dichloromethane (50.0 mL), followed by the sequential addition of 1,3-malonic acid (2.00 g, 19.2 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (11.1 g, 57.7 mmol), and 4-dimethylaminopyridine (470 mg, 3.84 mmol). The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to give dioctyl propylene glycol (5.00 g, 79.2% yield).

[0817] Step 2: Synthesis of dioctyl-2-(4-benzyloxybutyl)propanediol

[0818] Reaction formula:

[0819] Material proportions:

[0820] Operation process:

[0821] Dioctyl propylene glycol (5.00 g, 15.2 mmol) and 4-bromobutoxymethylbenzene (3.70 g, 15.2 mmol) were dissolved in acetonitrile (10.0 mL). Tetrabutylammonium bromide (491 mg, 1.52 mmol) and potassium carbonate (2.31 g, 16.7 mmol) were added sequentially. The reaction was carried out at 60 °C for 4 hours under nitrogen protection. TLC monitoring showed a large amount of product formation, indicating the reaction was complete. The mixture was filtered and concentrated. The crude product was then subjected to column chromatography to obtain a colorless liquid, dioctyl-2-(4-benzyloxybutyl)propylene glycol (3.200 g, 42.8% yield).

[0822] Step 3: Synthesis of dioctyl-2-(4-hydroxybutyl)propanediol

[0823] Reaction formula:

[0824] Material proportions:

[0825] Operation process:

[0826] Dioctyl-2-(4-benzyloxybutyl)propanediol (3.10 g, 6.32 mmol) was dissolved in methanol (30.0 mL), and palladium / carbon (0.672 g, 0.63 mmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. The crude product was evaporated to dryness and then subjected to column chromatography to obtain a colorless liquid dioctyl-2-(4-hydroxybutyl)propanediol (2.51 g, 99% yield).

[0827] Step 4: Synthesis of dioctyl-2-[4-(p-toluenesulfonyloxy)butyl]propanediol

[0828] Reaction formula:

[0829] Material proportions:

[0830] Operation process:

[0831] Dioctyl-2-(4-hydroxybutyl)propanediol (1.00 g, 2.50 mmol) was dissolved in dichloromethane (10.0 mL), followed by the sequential addition of triethylamine (757 mg, 7.49 mmol) and 4-dimethylaminopyridine (30.5 mg, 0.25 mmol). Finally, 4-methylbenzenesulfonyl chloride (713 mg, 3.74 mmol) was added dropwise at 0 °C under nitrogen protection. The reaction was carried out at 25 °C for 12 hours. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain dioctyl-2-[4-(p-toluenesulfonyloxy)butyl]propanediol (1.30 g, 93% yield).

[0832] Step 5: Synthesis of [3-(6-bromohexanoyloxy)-2-octaoxopropyl]octyl ester

[0833] Reaction formula:

[0834] Material proportions:

[0835] Operation process:

[0836] Dioctyl-2-[4-(p-toluenesulfonyloxy)butyl]propanediol (1.00 g, 1.80 mmol) was dissolved in acetonitrile (10.0 mL), and 1-octylnonyl-8-[(4-hydroxycyclohexyl)amino]octyl ester (447 mg, 0.91 mmol), potassium carbonate (338 mg, 2.25 mmol), sodium iodide (623 mg, 4.51 mmol), and tetrahydrofuran (3.00 mL) were added sequentially. The reaction was carried out at 85 °C for 20 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The resulting yellow liquid, [dioctyl-2-[4-[(4-hydroxycyclohexyl)-[8-(1-octylnonoxo)-8-oxoylide-octyl]amino]butyl]propanediol (70.0 mg, 9.00% yield), was purified by column chromatography.

[0837] 1H NMR (400MHz, CHLOROFORM-d) δ=10.02-9.79(m,1H),4.86(quin,J=6.2Hz,1H),4.19-4.05(m,5H),3.34(br t,J=7.2Hz,2H),3.18-2.85(m,4H),2.29(t,J=7.4Hz,2H),2.16-1.85(m,12H),1.71-1.40(m,32H),1.38-1.27(m,32H),0.95-0.79(m,12H)ppm.

[0838] LCMS: RT=1.949, m / z=879.2.[M+H] + .

[0839] Preparation Example 26: Preparation of Compound 24

[0840] Step 1: Preparation of 24-1

[0841] Reaction formula:

[0842] Material proportions:

[0843] Operation process:

[0844] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 18 g of oil.

[0845] Step 2: Preparation of 24-2

[0846] Reaction formula:

[0847] Material proportions:

[0848] Operation process:

[0849] 24-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at room temperature for 2 hours. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 3.4 g of oil.

[0850] Step 3: Preparation of Compound 24

[0851] Reaction formula:

[0852] Material proportions:

[0853] Operation process:

[0854] 24-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 20:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 1.8 g of oil.

[0855] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.2Hz,2H),4.21(dd,J=11.8,3.3Hz,2H),4.00(dd,J=11.8,6.9Hz,2H),3.56(dt,J=11.0,5. 9Hz,1H),2.30(td,J=7.5,2.1Hz,8H),2.03(t,J=6.9Hz,2H),1.86(s,1H),1.66–1.45(m,16H),1.38–1.17(m,50H),0.88(q,J=7.0Hz,12H).

[0856] MS(ES+)m / z): 907.9(M) + .

[0857] Preparation Example 27: Preparation of Compound 29

[0858] Step 1: Synthesis of (3-benzyloxy-2-octaoxo-propoxy)methylbenzene

[0859] Reaction formula:

[0860] Material proportions:

[0861] Operation process:

[0862] 1,3-Dibenzyloxypropane-2-ol (3.00 g, 11.0 mmol) and sodium hydride (661 mg, 16.5 mmol, 60%) were dissolved in N,N-dimethylformamide (30.0 mL). The mixture was stirred at 25 °C for 0.5 h under nitrogen protection. 1-Iodooctane (3.17 g, 13.2 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was carried out at 20 °C for 12 h under nitrogen protection. The reaction was monitored by TLC until completion. The reaction solution was extracted and concentrated. The crude product was purified by column chromatography to give a colorless liquid (3-benzyloxy-2-octaoxo-propoxy)methylbenzene (1.70 g, 40.0% yield).

[0863] Step 2: Synthesis of 2-octaoxopropane-1,3-diol

[0864] Reaction formula:

[0865] Material proportions:

[0866] Operation process:

[0867] 1.70 g (4.42 mmol) of 3-benzyloxy-2-octaoxo-propoxy)methylbenzene was dissolved in methanol (20.0 mL). Palladium / carbon (0.941 g, 0.88 mmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. The crude product was evaporated to dryness and then subjected to column chromatography to give a colorless liquid 2-octaoxopropane-1,3-diol (900 mg, 99% yield).

[0868] Step 3: Synthesis of (3-hydroxy-2-octaoxopropyl)octyl ester

[0869] Reaction formula:

[0870] Material proportions:

[0871] Operation process:

[0872] 2-Octaoxypropane-1,3-diol (900 mg, 4.41 mmol) was dissolved in dichloromethane (10.0 mL), and then octanoic acid (572 mg, 3.96 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.01 g, 5.29 mmol), and 4-dimethylaminopyridine (54.0 mg, 440 μmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid (3-hydroxy-2-octaoxypropyl)octyl ester (700 mg, 48.0% yield).

[0873] Step 4: Synthesis of [3-(6-bromohexanoyloxy)-2-octaoxopropyl]octyl ester

[0874] Reaction formula:

[0875] Material proportions:

[0876] Operation process:

[0877] (3-hydroxy-2-octaoxopropyl)octyl ester (700 mg, 2.12 mmol) was dissolved in dichloromethane (10.0 mL), and 6-bromohexanoic acid (620 mg, 3.18 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (609 mg, 3.18 mmol), and 4-dimethylaminopyridine (26.0 mg, 210 μmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid [3-(6-bromohexanoyloxy)-2-octaoxopropyl]octyl ester (520 mg, 48.3% yield).

[0878] Step 5: Synthesis of [3-[6-[(4-hydroxycyclohexyl)-[6-(3-octanoyloxy-2-octaoxo-propoxy)-6-oxylidene-hexyl]amino]hexanoyloxy]-2-octaoxopropyl]octyl ester

[0879] Reaction formula:

[0880] Material proportions:

[0881] Operation process:

[0882] [3-(6-bromohexanoyloxy)-2-octaoxopropyl]octyl ester (1.20 g, 2.36 mmol) was dissolved in acetonitrile (15.00 mL), followed by the addition of 4-aminocyclohexanol (123 mg, 1.06 mmol), potassium carbonate (980 mg, 7.09 mmol), potassium iodide (589 mg, 3.55 mmol), and tetrahydrofuran (5.00 mL). The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a colorless liquid [3-[6-[(4-hydroxycyclohexyl)-[6-(3-octanoyloxy-2-octaoxo-propoxy)-6-oxylidene-hexyl]amino]hexanoyloxy]-2-octaoxopropyl]octyl ester (700 mg, 30.0% yield).

[0883] 1H NMR (400MHz, CHLOROFORM-d) δ=9.92-9.77(m,1H),4.21-4.09(m,8H),3.69(t,J=5.2Hz,2H),3.59-3.48(m,4H),3.38(br t,J=8.4Hz,1H),3.20-2.93(m,4H),2.35(td,J1=7.4,J2=17.6Hz,8H),2.17- 1.93(m,10H),1.78-1.48(m,18H),1.41(td,J1=7.6,J2=15.4Hz,4H),1.28(br d,J=5.8Hz,33H),1.19-0.65(m,12H)ppm.

[0884] LCMS: RT=2.429, m / z=996.8.[M+H] + .

[0885] Preparation Example 28: Preparation of Compound 11

[0886] Step 1: Synthesis of 1,3-Di(benzyloxy)propane-2-yl-8-bromooctyl ester

[0887] Reaction formula:

[0888] Material proportions:

[0889] Operation process:

[0890] 1,3-Di(benzyloxy)propane-2-ol (24.00 g, 7.34 mmol) and 8-bromooctanoic acid (1.97 g, 8.81 mmol) were dissolved in dichloromethane (20 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.11 g, 11.02 mmol) and 4-dimethylaminopyridine (179 mg, 1.47 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1,3-di(benzyloxy)propane-2-yl 8-bromooctyl ester (3.40 g, 96.9% yield).

[0891] Step 2: Synthesis of 1,3-dihydroxypropane-2-yl-8-bromooctyl ester

[0892] Reaction formula:

[0893] Material proportions:

[0894] Operation process:

[0895] 1,3-Di(benzyloxy)propane-2-yl 8-bromooctyl ester (3.40 g, 7.12 mmol) was dissolved in methanol (34 mL), and palladium on carbon (1.52 g, 1.42 mmol) was added. The mixture was reacted at 35 °C and 40 Psi for 12 hours under hydrogen protection. The reaction was monitored by TLC until completion, and then filtered and concentrated. The resulting solution was purified by column chromatography to obtain a yellow liquid, 1,3-dihydroxypropane-2-yl 8-bromooctyl ester (1.34 g, 63.3% yield).

[0896] Step 3: Synthesis of 1,3-bis(hexanoyloxy)propane-2-yl 8-bromooctyl ester

[0897] Reaction formula:

[0898] Material proportions:

[0899] Operation process:

[0900] 1,3-Dihydroxypropane-2-yl 8-bromooctyl ester (1.34 g, 4.51 mmol) and hexanoic acid (1.05 g, 9.02 mmol) were dissolved in dichloromethane (15 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.59 g, 13.5 mmol) and 4-dimethylaminopyridine (220 mg, 1.80 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1,3-di(hexanoyloxy)propane-2-yl 8-bromooctyl ester (2.00 g, 68.7% yield).

[0901] Step 4: Synthesis of undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester

[0902] Reaction formula:

[0903] Material proportions:

[0904] Operation process:

[0905] Undecyl 6-bromohexyl ester (2.00 g, 5.72 mmol) was dissolved in acetonitrile (20 mL). (1s,4s)-4-aminocyclohexane-1-ol (1.32 g, 11.4 mmol) and triethylamine (1.32 g, 11.4 mmol) were added to the reaction solution. The reaction was carried out at 40 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the organic phase was concentrated. The crude product was purified by column chromatography to obtain a pink liquid undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (910 mg, 41.1% yield).

[0906] Step 5: Synthesis of 1,3-bis(hexanoyloxy)propane-2-yl 8-{[6-oxoylide-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester

[0907] Reaction formula:

[0908] Material proportions:

[0909] Operation process:

[0910] 1,3-Di(hexanoyloxy)propane-2-yl 8-bromooctyl ester (1.17 g, 2.36 mmol) was dissolved in acetonitrile (10 mL), and undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (755 mg, 1.97 mmol), potassium carbonate (952 mg, 6.89 mmol), potassium iodide (392 mg, 2.36 mmol), and tetrahydrofuran (2 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid, 1,3-di(hexanoyloxy)propane-2-yl 8-{[6-oxoylide-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester (300 mg, 70.4% yield).

[0911] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.30-5.23 (m, 1H), 4.30 (dd, J1 = 4.4, J2 = 12.0Hz, 2H),4.15(dd,J1=6.0,J2=12.0Hz,3H),4.06(t,J=6.8Hz,2H),3.45-3.34(m,1H), 3.17-3.06(m,2H),3.04-2.93(m,2H),2.36-2.30(m,8H),2.19-2.07(m,3H),2.05 -1.92(m,7H),1.74-1.60(m,11H),1.47-1.21(m,34H),0.90(q,J=6.8Hz,9H)ppm.

[0912] LCMS:RT=0.740,m / z 797.3[M+H] + .

[0913] Preparation Example 29: Preparation of Compound 16

[0914] Step 1: Synthesis of 4-nitrophenylpentadecan-8-yl carbonate

[0915] Reaction formula:

[0916] Material proportions:

[0917] Operation process:

[0918] Pentadecane-8-ol (1.00 g, 4.38 mmol) was dissolved in dichloromethane (10.0 mL), and 4-dimethylaminopyridine (1.07 g, 8.76 mmol) was added at 0 °C. Chlorinated 4-nitrophenyl (1.06 g, 5.25 mmol) was dissolved in dichloromethane (10.0 mL) and slowly added dropwise at 0 °C. After the addition was complete, the reaction was carried out at 25 °C for 12 hours. The reaction was monitored by TLC until it ended. The solution was used directly in the next step to obtain a yellow liquid 4-nitrophenylpentadecan-8-yl carbonate (1.72 g, 100% yield).

[0919] Step 2: Synthesis of 6-bromohexyl-3-(benzyloxy)propyl ester

[0920] Reaction formula:

[0921] Material proportions:

[0922] Operation process:

[0923] 6-Bromohexanol (1.91 g, 10.5 mmol) and 3-(benzyloxy)propionic acid (2.00 g, 11.1 mmol) were dissolved in dichloromethane (20.0 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.19 g, 16.6 mmol) and 4-dimethylaminopyridine (135 mg, 1.11 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the organic phase was concentrated. Purification by column chromatography yielded a colorless liquid, 6-bromohexyl-3-(benzyloxy)propyl ester (2.67 g, 70.0% yield).

[0924] Step 3: Synthesis of 6-bromohexyl-3-hydroxypropyl ester

[0925] Reaction formula:

[0926] Material proportions:

[0927] Operation process:

[0928] 6-Bromohexyl-3-(benzyloxy)propyl ester (2.10 g, 6.12 mmol) was dissolved in dichloromethane (21.0 mL), and boron trichloride (1 M, 12.2 mL) was added at 0 °C. The reaction was then carried out under nitrogen protection at 0 °C for 1 hour. TLC monitoring showed the formation of new spots. After adjusting the pH to neutral, the mixture was extracted with ethyl acetate, and the organic phase was concentrated. The solution was purified by column chromatography to obtain a colorless liquid 6-bromohexyl-3-hydroxypropyl ester (1.50 g, 96.8% yield).

[0929] Step 4: Synthesis of 6-bromohexyl 3-{[(pentadecan-8-oxy)carbonyl]oxo}propyl ester

[0930] Reaction formula:

[0931] Material proportions:

[0932] Operation process:

[0933] 6-Bromohexyl 3-hydroxypropyl ester (1.00 g, 3.95 mmol) was dissolved in dichloromethane (10.0 mL). 4-Nitrophenylpentadecan-8-yl carbonate (1.55 g, 3.95 mmol) was slowly added at 25 °C. After the addition was complete, the reaction was carried out at 25 °C for 12 hours. The reaction was monitored by TLC until it ended. The product was purified by column chromatography to obtain a colorless liquid 6-bromohexyl 3-{[(pentadecan-8-oxy)carbonyl]oxo}propyl ester (1.30 g, 64.8% yield).

[0934] Step 5: Synthesis of 6-({6-[(3-{[(pentadecan-8-oxy)carbonyl]oxo}propionyl)oxo]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl3-{[(pentadecan-8-oxy)carbonyl]oxo}propyl ester)

[0935] Reaction formula:

[0936] Material proportions:

[0937] Operation process:

[0938] 6-Bromohexyl 3-{[(pentadecano-8-oxy)carbonyl]oxo}propyl ester (940 mg, 1.85 mmol) was dissolved in acetonitrile (10.0 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (102 mg, 888 μmol), potassium carbonate (895 mg, 6.48 mmol), potassium iodide (368 mg, 2.22 mmol), and tetrahydrofuran (5.00 mL). The reaction was carried out at 75 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, then filtered and concentrated. Purified by column chromatography, the colorless liquid 6-({6-[(3-{[(pentadecan-8-oxy)carbonyl]oxo}propionyl)oxo]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl3-{[(pentadecan-8-oxy)carbonyl]oxo}propyl ester (163 mg, 9.09% yield) was obtained.

[0939] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.69 (quin, J = 6.2Hz, 2H), 4.40 (t, J = 6.6Hz, 4H), 4.10 (t, J = 6.8Hz, 4H), 4.04-3.96 (m, 1H), 2.70 (t, J = 6.6Hz, 4H), 2.44 (br s,3H),1.87(br s,3H),1.69-1.54(m,20H),1.43-1.13(m,52H),0.89(t,J=6.8Hz,12H)ppm.

[0940] LCMS:RT=2.490,m / z 969.6[M+H] + .

[0941] Preparation Example 30: Preparation of Compound 18

[0942] Step 1: Synthesis of 1-(benzyloxy)dodecane-2-ol

[0943] Reaction formula:

[0944] Material proportions:

[0945] Operation process:

[0946] 1-(benzyloxy)dodecane-2-ol (4.00 g, 11.0 mmol) and sodium hydride (656 mg, 16.4 mmol, 60%) were dissolved in N,N-dimethylformamide (50.0 mL). The mixture was stirred at 25 °C for 0.5 h under nitrogen protection. 1-Iodooctane (3.94 g, 13.2 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was carried out at 20 °C for 12 h under nitrogen protection. The reaction was monitored by TLC until completion. The reaction solution was extracted and concentrated. The crude product was subjected to column chromatography to give a colorless liquid 1-(benzyloxy)dodecane-2-ol (1.50 g, 34.0% yield).

[0947] Step 2: Synthesis of 2-(octoxy)dodecane-1-ol

[0948] Reaction formula:

[0949] Material proportions:

[0950] Operation process:

[0951] ({[2-(octoxy)dodecyl]oxo}methyl)benzene (1.50 g, 3.71 mmol) was dissolved in methanol (30.0 mL), and palladium / carbon (0.394 g, 0.37 mmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. The crude product was evaporated to dryness and then subjected to column chromatography to give a colorless liquid 2-(octoxy)dodecane-1-ol (1.17 g, 99.9% yield).

[0952] Step 3: Synthesis of 2-(octoxy)dodecyl 6-bromohexyl ester

[0953] Reaction formula:

[0954] Material proportions:

[0955] Operation process:

[0956] 2-(octoxy)dodecane-1-ol (1.17 g, 3.72 mmol) was dissolved in dichloromethane (10.0 mL), followed by the sequential addition of 6-bromohexanoic acid (1.09 g, 5.58 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.07 g, 5.58 mmol), and 4-dimethylaminopyridine (45.4 mg, 370 μmol). The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid, 2-(octoxy)dodecyl 6-bromohexyl ester (1.70 g, 93.0% yield).

[0957] Step 4: Synthesis of 2-(octoxy)dodecyl 6-[(6-{[2-(octoxy)dodecyl]oxo}-6-oxoylide)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl ester

[0958] Reaction formula:

[0959] Material proportions:

[0960] Operation process:

[0961] 2-(octoxy)dodecyl 6-bromohexyl ester (1.50 g, 3.05 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (158 mg, 1.37 mmol), potassium carbonate (1.27 g, 9.15 mmol), potassium iodide (759 mg, 4.58 mmol), and tetrahydrofuran (5 mL) were added sequentially. The reaction was carried out at 80 °C for 20 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid, 2-(octoxy)dodecyl 6-[(6-{[2-(octoxy)dodecyl]oxo}-6-oxoylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl ester (700 mg, 24.15%).

[0962] 1 H NMR(400MHz, CHLOROFORM-d)δ=4.16-4.08(m,3H),4.06-3.99(m,2H),3.55(td,J1=6.4,J2=9.2Hz,2H),3.46-3.39(m,4H),3.34-3.23(m,1H),3.10- 2.99(m,2H),2.96-2.85(m,2H),2.36(t,J=7.2Hz,4H),2.07-1.91(m,10H) ,1.69(quin,J=7.6Hz,4H),1.58-1.52(m,5H),1.50-1.45(m,3H),1.41(br dd,J=7.6,15.6Hz,59H),0.89(t,J=6.8Hz,12H).

[0963] LCMS:RT=2.469,m / z 937.9[M+H] + .

[0964] Preparation Example 31: Preparation of Compound 31

[0965] Step 1: Synthesis of 6-[(6-{[2-(octanoyloxy)dodecanoyl]oxo}hexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl 2-(octanoyloxy)dodecane ester

[0966] Reaction formula:

[0967] Material proportions:

[0968] Operation process:

[0969] 6-Bromohexyl 2-(octanoyloxy)dodecane ester (1.50 g, 2.97 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (153 mg, 1.34 mmol), potassium carbonate (1.23 g, 8.90 mmol), potassium iodide (738 mg, 4.45 mmol), and tetrahydrofuran (5 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid, 6-[(6-{[2-(octanoyloxy)dodecanoyl]oxo}hexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl 2-(octanoyloxy)dodecane ester (580 mg, 20.2% yield).

[0970] 1 H NMR (400MHz, CHLOROFORM-d) δ = 11.57-11.38 (m, 1H), 4.96 (t, J = 6.4Hz, 2H), 4. 23-4.09(m,5H),3.82-3.72(m,3H),3.36-3.26(m,1H),3.14-3.00(m,2H),2.98 -2.85(m,2H),2.46-2.36(m,4H),2.12-1.91(m,10H),1.90-1.78(m,7H),1.73 -1.64(m,9H),1.46-1.38(m,12H),1.36-1.23(m,43H),0.99-0.79(m,12H)ppm.

[0971] LCMS:RT=2.217,m / z 964.8[M+H] + .

[0972] Preparation Example 32: Preparation of Compound 32

[0973] Step 1: Synthesis of ({[1-(benzyloxy)-3-[(6-bromohexyl)oxo]propane-2-yl]oxo}methyl)benzene

[0974] Reaction formula:

[0975] Material proportions:

[0976] Operation process:

[0977] 2,3-Di(benzyloxy)propane-1-ol (900 mg, 3.30 mmol) was dissolved in N,N-dimethylformamide (30 mL), and sodium hydrogen (60%) (198 mg, 4.96 mmol) was added at 0 °C. The reaction was continued at 0 °C for 0.5 h, followed by the addition of 6-bromohexane-1-ol (1.21 g, 4.96 mmol). The reaction was continued at 25 °C for 12 h under nitrogen protection. TLC monitoring showed the formation of new spots. The reaction solution was quenched with 300 mL of water, washed twice with 300 mL of ethyl acetate each time, and the organic phase was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid ({[1-(benzyloxy)-3-[(6-bromohexyl)oxo]propane-2-yl]oxo}methyl)benzene (740 mg, 51.4% yield).

[0978] Step 2: Synthesis of 3-[(6-bromohexyl)oxo]propane-1,2-diol

[0979] Reaction formula:

[0980] Material proportions:

[0981] Operation process:

[0982] ({[1-(benzyloxy)-3-[(6-bromohexyl)oxo]propane-2-yl]oxo}methyl)benzene (740 mg, 1.70 mmol) was dissolved in dichloromethane (8 mL), and boron trichloride, 1 M CH2Cl2 solution (597 mg, 5.10 mmol) was added at 0 °C. The reaction was carried out at 0 °C for 1 hour. The reaction was monitored by TLC until it ended. The solution was quenched with 10 mL of sodium bicarbonate aqueous solution, and extracted twice with 10 mL of dichloromethane each time. The organic phase was dried, filtered, and concentrated. The solution was purified by column chromatography to obtain a colorless liquid 3-[(6-bromohexyl)oxo]propane-1,2-diol (240 mg, 55.3% yield).

[0983] Step 3: Synthesis of 1-[(6-bromohexyl)oxo]-3-(octanoyloxo)propane-2-yloctyl ester

[0984] Reaction formula:

[0985] Material proportions:

[0986] Operation process:

[0987] 240 mg (0.940 mmol) of 3-[(6-bromohexyl)oxo]propane-1,2-diol and octanoic acid (298 mg, 2.07 mmol) were dissolved in 3 mL of dichloromethane. 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (540 mg, 2.82 mmol) and 4-dimethylaminopyridine (45.9 mg, 0.376 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-[(6-bromohexyl)oxo]-3-(octanoyloxo)propane-2-yloctyl ester (380 mg, 79.60% yield).

[0988] Step 4: Synthesis of 1-{[6-({6-[2,3-di(octanoyloxy)propoxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl]oxo}-3-(octanoyloxy)propane-2-yloctyl ester

[0989] Reaction formula:

[0990] Material proportions:

[0991] Operation process:

[0992] 1-[(6-bromohexyl)oxo]-3-(octanoyloxo)propane-2-yloctyl ester (300 mg, 0.591 mmol) was dissolved in acetonitrile (2 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (30.6 mg, 0.265 mmol), potassium carbonate (245 mg, 1.77 mmol), potassium iodide (147 mg, 0.886 mmol), and tetrahydrofuran (1 mL). The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid 1-{[6-({6-[2,3-di(octanoyloxo)propoxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl]oxo}-3-(octanoyloxo)propane-2-yloctyl ester (130 mg, 22.4% yield).

[0993] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.26-5.15 (m, 2H), 4.34 (dd, J1 = 3.6, J2 = 12.0Hz, 2H), 4.16 (dd, J1 = 6.4, J2 = 12.0Hz, 2H), 4.06-3.96 (m ,1H),3.57-3.49(m,4H),3.47-3.38(m,4H),2.54-2.36(m,3H),2.32(q,J=7.2Hz,8H),1.96-1.79(m,3H),1.69-1.50(m,21H),1.30(br s,43H),0.96-0.83(m,12H)ppm.

[0994] LCMS:RT=1.643,m / z 968.8[M+H] + .

[0995] Preparation Example 34: Preparation of Compound 53

[0996] Step 1: Synthesis of 2-hydroxydodecyl 6-bromohexyl ester

[0997] Reaction formula:

[0998] Material proportions:

[0999] Operation process:

[1000] 6-Bromohexanoic acid (4.66 g, 23.8 mmol), ferric chloride (88.0 mg, 0.542 mmol), and pyridine (48.0 mg, 0.607 mmol) were added to 2-decyloxapropylcyclohexane (4.00 g, 21.7 mmol). The reaction was carried out at 40 °C for 12 h under nitrogen protection. TLC monitoring showed the formation of new spots, and the organic phase was concentrated. The crude product was purified by column chromatography to give a colorless liquid 2-hydroxydodecyl 6-bromohexyl ester (6.40 g, 77.7% yield).

[1001] Step 2: Synthesis of 2-{[5-(1,2-dithiapentan-3-yl)valeryl]oxo}dodecyl 6-bromohexyl ester

[1002] Reaction formula:

[1003] Material proportions:

[1004] Operation process:

[1005] 2-Hydroxydodecyl 6-bromohexyl ester (3.00 g, 7.91 mmol) and 5-(1,2-dithiapentylcyclo-3-yl)valeric acid (1.63 g, 7.91 mmol) were dissolved in dichloromethane (30 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27 g, 11.8 mmol) and 4-dimethylaminopyridine (193 mg, 1.58 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a yellow liquid, 2-{[5-(1,2-dithiapentylcyclo-3-yl)valeryl]oxo}dodecyl 6-bromohexyl ester (2.7 g, 60.15% yield).

[1006] Step 3: Synthesis of 2-{[5-(1,2-dithiapentyl-3-yl)valeryl]oxo}dodecyl 6-({6-[(2-{[5-(1,2-dithiapentyl-3-yl)valeryl]oxo}dodecyl)oxo]-6-oxoylidexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[1007] Reaction formula:

[1008] Material proportions:

[1009] Operation process:

[1010] 2-{[5-(1,2-dithiapentylcyclo-3-yl)pentanoyl]oxo}dodecyl 6-bromohexyl ester (1.20 g, 2.11 mmol) was dissolved in acetonitrile (10 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (109 mg, 0.951 mmol), potassium carbonate (876 mg, 6.34 mmol), potassium iodide (526 mg, 3.17 mmol), and tetrahydrofuran (2 mL). The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, then filtered and concentrated. The yellow liquid 2-{[5-(1,2-dithiapentyl-3-yl)valeryl]oxo}dodecyl 6-({6-[(2-{[5-(1,2-dithiapentyl-3-yl)valeryl]oxo}dodecyl)oxo]-6-oxoylide hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (430 mg, 17.36% yield) was purified by column chromatography.

[1011] 1H NMR(400MHz,CHLOROFORM-d)δ=5.16-5.03(m,2H),4.33-4.23(m,2H),4.13(br s,1H),4.00(ddd,J1=4.8,J2=6.8,J3=12.0Hz,2H),3.80-3.70(m,3H),3.64-3.53(m, 2H),3.44-3.33(m,1H),3.24-3.09(m,5H),3.05-2.93(m,2H),2.54-2.43(m,2H),2.3 5(dt,J1=2.8,J2=7.2Hz,8H),2.14-1.90(m,11H),1.86(td,J1=3.2,J2=6.8Hz,3H),1 .77-1.62(m,13H),1.53-1.38(m,9H),1.36-1.19(m,33H),0.89(t,J=6.8Hz,6H)ppm.

[1012] LCMS:RT=2.441,m / z 1089.1[M+H] + .

[1013] Preparation Example 35: Preparation of Compound 54

[1014] Step 1: Synthesis of 3-(Butylmercapto)propionic acid

[1015] Reaction formula:

[1016] Material proportions:

[1017] Operation process:

[1018] 3-Mercaptopropionic acid (4.00 g, 37.6 mmol) and 1-iodobutane (6.93 g, 37.6 mmol) were dissolved in methanol (40 mL), and potassium hydroxide (6.34 g, 45.2 mmol) was added. The reaction was carried out at 20 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots. 10 mL of water was added, and the mixture was extracted with ethyl acetate. The pH of the aqueous phase was adjusted to 5 with 1 M hydrochloric acid, and the mixture was extracted twice with 20 mL of ethyl acetate each time. The organic phase was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid 3-(butylmercapto)propionic acid (2.13 g, 34.8% yield).

[1019] Step 2: Synthesis of 2-{[3-(butylmercapto)propionyl]oxo}dodecyl 6-bromohexyl ester

[1020] Reaction formula:

[1021] Material proportions:

[1022] Operation process:

[1023] 2-Hydroxydodecyl 6-bromohexyl ester (3.00 g, 7.91 mmol) and 3-(butylmercapto)propionic acid (1.54 g, 9.49 mmol) were dissolved in dichloromethane (30 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27 g, 11.8 mmol) and 4-dimethylaminopyridine (193 mg, 1.58 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid, 2-{[3-(butylmercapto)propionyl]oxo}dodecyl 6-bromohexyl ester (3.14 g, 75.8% yield).

[1024] Step 3: Synthesis of 2-{[3-(butylmercapto)propionyl]oxo}dodecyl 6-({6-[(2-{[3-(butylmercapto)propionyl]oxo}dodecyl)oxo]-6-oxoylide}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[1025] Reaction formula:

[1026] Material proportions:

[1027] Operation process:

[1028] 2-{[3-(butylmercapto)propionyl]oxo}dodecyl 6-bromohexyl ester (1.50 g, 2.86 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (148 mg, 1.29 mmol), potassium carbonate (1.19 g, 8.59 mmol), potassium iodide (713 mg, 4.30 mmol), and tetrahydrofuran (5 mL) were added sequentially. Finally, the reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, and then filtered and concentrated. The yellow liquid 2-{[3-(butylmercapto)propionyl]oxo}dodecyl 6-({6-[(2-{[3-(butylmercapto)propionyl]oxo}dodecyl)oxo]-6-oxoylide}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (600 mg, 20.1% yield) was purified by column chromatography.

[1029] 1H NMR(400MHz,CHLOROFORM-d)δ=5.14-5.04(m,2H),4.30-4.22(m,2H),4.11(br s,1H),4.07-3.97(m,2H),3.78-3.73(m,1H),3.36-3.23(m,1H),3.13-3.01(m,2H),2.99-2.87(m,2H),2. 81-2.74(m,4H),2.65-2.58(m,4H),2.57-2.50(m,4H),2.35(t,J=7.2Hz,4H),2.11-2.03(m,3H),1.99(br d,J=8.8Hz,7H),1.86(td,J1=3.2,J2=6.8Hz,1H),1.71-1.65(m,4H),1.6 0-1.54(m,8H),1.45-1.38(m,8H),1.26(s,33H),0.97-0.84(m,12H)ppm.

[1030] LCMS:RT=2.477,m / z 1001.2[M+H] + .

[1031] Preparation Example 36: Preparation of Compound 33

[1032] Step 1: Preparation of 33-1

[1033] Reaction formula:

[1034] Material proportions:

[1035] Operation process:

[1036] 6-Bromohexanoic acid, 1,2-epoxytetradecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1037] Step 2: Preparation of 33-2

[1038] Reaction formula:

[1039] Material proportions:

[1040] Operation process:

[1041] 33-1, n-decanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product Rf value was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 200 mL of saturated sodium bicarbonate aqueous solution, and then once with 200 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 10 g of oil.

[1042] Step 3: Preparation of Compound 33

[1043] Reaction formula:

[1044] Material proportions:

[1045] Operation process:

[1046] 33-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.2 g of oil.

[1047] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.8,3.3Hz,2H),4.21(dd,J=11.8,3.3Hz,2H),4.01(dd,J=11.8,6.8Hz,3H),2.57( s,3H),2.30(td,J=7.5,3.2Hz,8H),1.87(d,J=13.2Hz,2H),1.75–1.40(m,20H),1.36–1.19(m,70H),0.87(t,J=6.7Hz,12H).

[1048] MS(ES+)m / z): 1076.8 (M) + .

[1049] Preparation Example 37: Preparation of Compound 34

[1050] Step 1: Preparation of 34-1

[1051] Reaction formula:

[1052] Material proportions:

[1053] Operation process:

[1054] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1055] Step 2: Preparation of 34-2

[1056] Reaction formula:

[1057] Material proportions:

[1058] Operation process:

[1059] 34-1, nonanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product had an Rf value of 0.6 and a TLC (PE:EA = 10:1). The reaction solution was washed once with 200 mL of saturated sodium bicarbonate aqueous solution, and then once with 200 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 10 g of oil.

[1060] Step 3: Preparation of Compound 34

[1061] Reaction formula:

[1062] Material proportions:

[1063] Operation process:

[1064] 34-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.2 g of oil.

[1065] 1H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.9,3.2Hz,2H),4.22(dd,J=11.8,3.3Hz,2H),4.07(s,1H),4.00(dd,J=11.8,7.0Hz,2H) ,2.84(s,3H),2.31(q,J=7.3Hz,8H),2.04–1.71(m,8H),1.63(dt,J=13.5,7.2Hz,16H),1.41–1.20(m,58H),0.87(t,J=6.7Hz,12H).

[1066] MS(ES+)m / z): 992.7 (M) + .

[1067] Preparation Example 38 Preparation of Compound 36

[1068] Step 1: Synthesis of 1-[(6-{[6-({2-[(7-methyloctanoyl)oxo]dodecyl}oxo)-6-oxylidenehexyl][(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxo]dodecane-2-yl7-methyloctyl ester]

[1069] Reaction formula:

[1070] Material proportions:

[1071] Operation process:

[1072] 1-[(6-bromohexanoyl)oxo]undecane-2-yloctyl ester (1.30 g, 2.50 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (129 mg, 1.13 mmol), potassium carbonate (1.04 g, 7.15 mmol), potassium iodide (622 mg, 3.75 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid, 1-[(6-{[6-({2-[(7-methyloctanoyl)oxo]dodecyl}oxo)-6-oxylidenehexyl][(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxo]dodecane-2-yl7-methyloctyl ester (750 mg, 30.1% yield).

[1073] 1H NMR(400MHz,CHLOROFORM-d)δ=5.13-5.03(m,2H),4.29-4.21(m,2H),4.10(br s,1H),4.06-3.95(m,2H),3.75(t,J=6.4Hz,2H),3.35-3.23(m,1H),3.12-2.99(m,2H),2.96-2.85(m,2H),2.34(br t,J=7.2Hz,6H),2.17-2.10(m,2H),2.08-1.95(m,10H),1.89-1.83(m,2H),1.72-1.63(m,5H),1.56(br s,4H),1.42-1.36(m,4H),1.34-1.21(m,34H),1.16-1.10(m,2H),0.99(d,J=6.4Hz,6H),0.95-0.83(m,24H)ppm.

[1074] LCMS:RT=2.777,m / z 993.5[M+H] + .

[1075] Preparation Example 39: Preparation of Compound 37

[1076] Step 1: Preparation of 37-1

[1077] Reaction formula:

[1078] Material proportions:

[1079] Operation process:

[1080] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1081] Step 2: Preparation of 37-2

[1082] Reaction formula:

[1083] Material proportions:

[1084] Operation process:

[1085] 37-1, hexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at -5℃ to 0℃ for 16 h. The Rf value of the product was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 200 mL of saturated sodium bicarbonate aqueous solution, and then once with 200 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 9 g of oil.

[1086] Step 3: Preparation of Compound 37

[1087] Reaction formula:

[1088] Material proportions:

[1089] Operation process:

[1090] 37-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.2 g of oil.

[1091] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.8,3.2Hz,2H),4.22(dd,J=11.8,3.3Hz,2H),4.00(dd,J=11.9,7.0Hz,3H),2.66( s,3H),2.30(td,J=7.5,3.9Hz,8H),1.89(d,J=13.9Hz,2H),1.81–1.45(m,22H),1.38–1.19(m,46H),0.88(q,J=7.0Hz,12H).

[1092] MS(ES+)m / z): 908.6 (M) + .

[1093] Preparation Example 40: Preparation of Compound 38

[1094] Step 1: Synthesis of 1-({6-[(6-{[2-(octanoyloxo)undecyl]oxo}-6-oxylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexanoyl}oxo)undecyl-2-yloctyl ester)

[1095] Reaction formula:

[1096] Material proportions:

[1097] Operation process:

[1098] 1-[(6-bromohexanoyl)oxo]undecane-2-yloctyl ester (1.00 g, 2.03 mmol) was dissolved in acetonitrile (7 mL), and (1s,4s)-4-aminocyclohexane-1-ol (105 mg, 0.915 mmol), potassium carbonate (843 mg, 6.10 mmol), potassium iodide (506 mg, 3.05 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid 1-({6-[(6-{[2-(octanoyloxo)undecyl]oxo}-6-oxylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexanoyl}oxo)undecane-2-yloctyl ester (540 mg, 28.3% yield).

[1099] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.14-5.02(m,2H),4.29-4.20(m,2H),4.11(br s,1H),4.04-3.94(m,2H),3.77-3.74(m,4H),3.36-3.22(m,1H),3.12-2.98(m,2H),2.95-2.84(m,2H),2.32(td,J1=7.6,J2=10.0Hz,8H) ,2.08-2.03(m,2H),2.02-1.92(m,7H),1.86(td,J1=3.2,J2=6.8Hz,4H),1.70-1.61(m,7H),1.44-1.20(m,49H),0.95-0.81(m,12H)ppm.

[1100] LCMS:RT=2.583,m / z 937.5[M+H] + .

[1101] Preparation Example 41: Preparation of Compound 39

[1102] Step 1: Preparation of 39-1

[1103] Reaction formula:

[1104] Material proportions:

[1105] Operation process:

[1106] 6-Bromohexanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 7 g of oil.

[1107] Step 2: Preparation of 39-2

[1108] Reaction formula:

[1109] Material proportions:

[1110] Operation process:

[1111] 39-1, n-decanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product Rf value was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 150 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7 g of oil.

[1112] Step 3: Preparation of Compound 39

[1113] Reaction formula:

[1114] Material proportions:

[1115] Operation process:

[1116] 39-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2 g of oil.

[1117] 1H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.8,3.3Hz,2H),4.22(dd,J=11.8,3.3Hz,2H),4.01(dt,J=11.8,6.3Hz,3H),2.91–2.57(m,3H) ,2.30(td,J=7.4,5.2Hz,8H),1.90(d,J=13.8Hz,2H),1.84–1.70(m,4H),1.68–1.51(m,16H),1.39–1.18(m,56H),0.87(t,J=6.7Hz,12H).

[1118] MS(ES+)m / z): 964.7 (M) + .

[1119] Preparation Example 42: Preparation of Compound 40

[1120] Step 1: Preparation of 40-1

[1121] Reaction formula:

[1122] Material proportions:

[1123] Operation process:

[1124] 6-Bromohexanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 7 g of oil.

[1125] Step 2: Preparation of 40-2

[1126] Reaction formula:

[1127] Material proportions:

[1128] Operation process:

[1129] 40-1, n-heptanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at -5℃ to 0℃ for 16 h. The Rf value of the product was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 150 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7 g of oil.

[1130] Step 3: Preparation of Compound 40

[1131] Reaction formula:

[1132] Material proportions:

[1133] Operation process:

[1134] 40-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2 g of oil.

[1135] 1 H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.8,3.3Hz,2H),4.22(dd,J=11.8,3.3Hz,2H),4.09–3.95(m,3H),2.80(s,3H ),2.31(td,J=7.4,5.5Hz,8H),1.92(d,J=14.1Hz,8H),1.69–1.49(m,16H),1.40–1.18(m,42H),0.87(h,J=3.7Hz,12H).

[1136] MS(ES+)m / z): 880.6(M) + .

[1137] Preparation Example 43: Preparation of Compound 41

[1138] Step 1: Synthesis of 1-({7-[(7-{[2-(octanoyloxo)undecyl]oxo}-7-oxylideneheptyl)[(1s,4s)-4-hydroxycyclohexyl]amino]heptanoyl}oxo)undecyl-2-yloctyl ester)

[1139] Reaction formula:

[1140] Material proportions:

[1141] Operation process:

[1142] 1-[(7-bromoheptanoyl)oxo]undecane-2-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (7 mL), and (1s,4s)-4-aminocyclohexane-1-ol (102 mg, 0.890 mmol), potassium carbonate (820 mg, 5.93 mmol), potassium iodide (492 mg, 2.97 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid 1-({7-[(7-{[2-(octanoyloxo)undecyl]oxo}-7-oxylideneheptyl)[(1s,4s)-4-hydroxycyclohexyl]amino]heptanoyl}oxo)undecane-2-yloctyl ester (540 mg, 28.2% yield).

[1143] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.6, J2 = 6.8Hz, 2H), 4.27-4.19 (m, 2H), 4.10 (br s, 1H), 4.05-3.97 (m, 2H), 3.77-3.74 (m, 2H), 3.29 (br t,J=9.6Hz,1H),3.09-2.98(m,2H),2.96-2.85(m,2H),2.31(t,J=7.6Hz,8H),2.06(br dd, J1=1.6, J2=9.2Hz,2H),2.03-1.89(m,8H),1.88-1.85(m,2H),1.67-1.59(m,10H),1.43-1.20(m,53H),0.94-0.82(m,12H)ppm.

[1144] LCMS:RT=2.625,m / z 965.6[M+H] + .

[1145] Preparation Example 44: Preparation of Compound 42

[1146] Step 1: Preparation of 42-1

[1147] Reaction formula:

[1148] Material proportions:

[1149] Operation process:

[1150] 7-Bromoheptanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1151] Step 2: Preparation of 42-2

[1152] Reaction formula:

[1153] Material proportions:

[1154] Operation process:

[1155] 42-1, n-heptanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product Rf value was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 150 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 8 g of oil.

[1156] Step 3: Preparation of Compound 42

[1157] Reaction formula:

[1158] Material proportions:

[1159] Operation process:

[1160] 42-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.4 g of oil.

[1161] 1H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.7,3.1Hz,2H),4.22(dd,J=11.8,3.5Hz,2H),4.09(s,1H),3.99(dd,J=11.8,6.9Hz,2H),3.33(s ,1H),3.17–2.86(m,4H),2.29(t,J=7.3Hz,8H),2.11–1.80(m,10H),1.65–1.48(m,14H),1.40–1.19(m,44H),0.86(dd,J=6.9,4.1Hz,12H).

[1162] MS(ES+)m / z): 908.7 (M) + .

[1163] Preparation Example 45: Preparation of Compound 43

[1164] Step 1: Preparation of 43-1

[1165] Reaction formula:

[1166] Material proportions:

[1167] Operation process:

[1168] 8-Bromooctanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1169] Step 2: Preparation of 43-2

[1170] Reaction formula:

[1171] Material proportions:

[1172] Operation process:

[1173] 43-1, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product Rf value was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 200 mL of saturated sodium bicarbonate aqueous solution, and then once with 200 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 10 g of oil.

[1174] Step 3: Preparation of Compound 43

[1175] Reaction formula:

[1176] Material proportions:

[1177] Operation process:

[1178] 43-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 20:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.1 g of oil.

[1179] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.2Hz,2H),4.21(dd,J=11.8,3.3Hz,2H),4.02(dd,J=11.9,6.7Hz,3H),2. 45(s,3H),2.29(t,J=7.5Hz,8H),1.85(d,J=13.1Hz,2H),1.72–1.43(m,20H),1.38–1.19(m,68H),0.87(t,J=6.6Hz,12H).

[1180] MS(ES+)m / z): 1048.8 (M) + .

[1181] Preparation Example 46: Preparation of Compound 44

[1182] Step 1: Synthesis of 2-(octanoyloxo)undecyl 8-[(8-{[2-(octanoyloxo)undecyl]oxo}-8-oxoylidecyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1183] Reaction formula:

[1184] Material proportions:

[1185] Operation process:

[1186] 2-(octanoyloxo)undecyl 8-bromooctyl ester (1.00 g, 1.92 mmol) was dissolved in acetonitrile (7 mL), and (1s,4s)-4-aminocyclohexane-1-ol (99.7 mg, 0.866 mmol), potassium carbonate (797 mg, 5.77 mmol), potassium iodide (479 mg, 2.89 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a yellow liquid, 2-(octanoyloxo)undecyl 8-[(8-{[2-(octanoyloxo)undecyl]oxo}-8-oxoylidecyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (510 mg, 26.7% yield).

[1187] 1 H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.6, J2 = 6.8Hz, 2H), 4.23 (dd, J1 = 3.6, J2 = 12.0Hz, 2H ),4.13-4.08(m,1H),4.05-3.97(m,2H),3.77-3.73(m,1H),3.34-3.22(m,1H),3.08-2.97(m,2H) ,2.95-2.84(m,2H),2.31(dt,J1=2.4,J2=7.6Hz,8H),2.09-2.02(m,2H),2.02-1.87(m,8H),1.87 -1.83(m,1H),1.67-1.62(m,5H),1.59-1.53(m,7H),1.41-1.22(m,57H),0.94-0.83(m,12H)ppm.

[1188] LCMS:RT=2.676,m / z 993.6[M+H] + .

[1189] Preparation Example 47: Preparation of Compound 45

[1190] Step 1: Preparation of 45-1

[1191] Reaction formula:

[1192] Material proportions:

[1193] Operation process:

[1194] 8-Bromooctanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1195] Step 2: Preparation of 45-2

[1196] Reaction formula:

[1197] Material proportions:

[1198] Operation process:

[1199] 45-1, octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask, and the mixture was stirred at -5℃ to 0℃ for 16 h. The Rf value of the product was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 150 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 8 g of oil.

[1200] Step 3: Preparation of Compound 45

[1201] Reaction formula:

[1202] Material proportions:

[1203] Operation process:

[1204] 45-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.5 g of oil.

[1205] 1H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.7,3.3Hz,2H),4.21(dd,J=11.8,3.3Hz,2H),4.10(q,J=2.7Hz,1H),4.00(dd,J=11.8,6.9Hz,2H),3.35(d ,J=11.7Hz,1H),3.16–2.87(m,4H),2.29(t,J=7.5Hz,8H),2.11–1.75(m,1 0H),1.65–1.50(m,14H),1.38–1.21(m,52H),0.86(td,J=7.0,1.9Hz,12H).

[1206] MS(ES+)m / z): 964.7 (M) + .

[1207] Preparation Example 48 Preparation of Compound 46

[1208] Step 1: Preparation of 46-1

[1209] Reaction formula:

[1210] Material proportions:

[1211] Operation process:

[1212] 8-Bromooctanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 8 g of oil.

[1213] Step 2: Preparation of 46-2

[1214] Reaction formula:

[1215] Material proportions:

[1216] Operation process:

[1217] 46-1, n-heptanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product Rf value was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 150 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 8 g of oil.

[1218] Step 3: Preparation of Compound 46

[1219] Reaction formula:

[1220] Material proportions:

[1221] Operation process:

[1222] 46-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.5 g of oil.

[1223] 1 H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.8,3.3Hz,2H),4.21(dd,J=11.8,3.3Hz,2H),4.11(q,J=2.7Hz,1H),4.00(dd,J=11.8,6.9Hz,2H),3.35(d,J =11.7Hz,1H),3.02(d,J=50.1Hz,4H),2.29(td,J=7.5,1.8Hz,8H),2.11–1. 73(m,10H),1.65–1.49(m,14H),1.37–1.21(m,48H),0.87(h,J=3.7Hz,12H).

[1224] MS(ES+)m / z): 936.7 (M) + .

[1225] Preparation Example 49: Preparation of Compound 35

[1226] Step 1: Synthesis of ({[5-(pentane-3-oxy)pentyl]oxo}methyl)benzene

[1227] Reaction formula:

[1228] Material proportions:

[1229] Five parallel reactions, procedure as follows:

[1230] 5-(benzyloxy)pentan-1-ol (500 mg, 2.57 mmol) and pentan-3-one (332 mg, 3.86 mmol) were dissolved in 1,2-dichloroethane (5.00 mL). Triethylsilane (448 mg, 3.86 mmol) and ytterbium trifluoromethanesulfonate (15.9 mg, 25.7 μmol) were added. The reaction was carried out at 92 °C for 48 hours under nitrogen protection. The reaction was stopped by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid ({[5-(pentan-3-oxy)pentyl]oxo}methyl)benzene (1.25 g, 36.7% yield).

[1231] Step 2: Synthesis of 5-(pentan-3-oxy)pentan-1-ol

[1232] Reaction formula:

[1233] Material proportions:

[1234] Operation process:

[1235] ({[5-(pentane-3-oxy)pentyl]oxo}methyl)benzene (1.25 g, 4.73 mmol) was dissolved in methanol (15.0 mL), and palladium / carbon (1.01 g, 945 μmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. The solution was evaporated to dryness to give a colorless liquid 5-(pentane-3-oxy)pentane-1-ol (732 mg, 88.8% yield).

[1236] Step 3: Synthesis of 5-(pentane-3-oxy)valerate

[1237] Reaction formula:

[1238] Material proportions:

[1239] Operation process:

[1240] 5-(pentane-3-oxy)pentane-1-ol (632 mg, 3.63 mmol) was dissolved in acetonitrile (7.00 mL) and water (3.50 mL). Diacetoxyiodobenzene (2.57 g, 7.98 mmol) was added, followed by 2,2,6,6-tetramethylpiperidine oxide (114 mg, 725 μmol). The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. The reaction was monitored by TLC until completion. The reaction solution was extracted with water and ethyl acetate and concentrated. Column chromatography yielded a colorless liquid, 5-(pentane-3-oxy)pentanoic acid (538 mg, 70.8% yield).

[1241] Step 4: Synthesis of 1-(benzyloxy)dodecane-2-yl5-(pentane-3-oxy)pentyl ester

[1242] Reaction formula:

[1243] Material proportions:

[1244] Operation process:

[1245] 1-(benzyloxy)dodecane-2-ol (0.92 g, 3.15 mmol) and 5-(pentane-3-oxy)valerate (592 mg, 3.15 mmol) were dissolved in dichloromethane (10.0 mL), followed by the sequential addition of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (904 mg, 4.72 mmol) and 4-dimethylaminopyridine (38.4 mg, 314 μmol). The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid, 1-(benzyloxy)dodecane-3-yloctyl ester (1.40 g, 96.4% yield).

[1246] Step 5: Synthesis of 1-hydroxydodecane-2-yl 5-(pentane-3-oxy)pentyl ester

[1247] Reaction formula:

[1248] Material proportions:

[1249] Operation process:

[1250] 1-(benzyloxy)dodecane-2-yl 5-(pentane-3-oxy)pentyl ester (1.40 g, 3.03 mmol) was dissolved in methanol (20.0 mL), and palladium / carbon (643 mg, 605 μmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. The solution was then evaporated to dryness to give a colorless liquid, 1-hydroxydodecane-2-yl 5-(pentane-3-oxy)pentyl ester (460 mg, 40.8% yield).

[1251] Step Six: Synthesis of 2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl 6-bromohexyl ester

[1252] Reaction formula:

[1253] Material proportions:

[1254] Operation process:

[1255] 1-Hydroxydodecane-2-yl 5-(pentane-3-oxy)pentyl ester (460 mg, 1.23 mmol) was dissolved in dichloromethane (10.0 mL), followed by the addition of 6-bromohexanoic acid (337 mg, 1.73 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (355 mg, 1.85 mmol), and 4-dimethylaminopyridine (30.1 mg, 246 μmol). The reaction was carried out under nitrogen protection at 25 °C for 12 hours. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid 2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl 6-bromohexyl ester (619 mg, 91.2% yield).

[1256] Step 7: Synthesis of 2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl 6-({6-oxoylide-6-[(2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl)oxo]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[1257] Reaction formula:

[1258] Material proportions:

[1259] Operation process:

[1260] 2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl 6-bromohexyl ester (519 mg, 944 μmol) was dissolved in acetonitrile (6.00 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (52.2 mg, 453 μmol), potassium carbonate (456 mg, 3.31 mmol), potassium iodide (188 mg, 1.13 mmol), and tetrahydrofuran (3.00 mL). The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, then filtered and concentrated. The yellow liquid 2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl 6-({6-oxoylide-6-[(2-{[5-(pentane-3-oxy)pentanoyl]oxo}dodecyl)oxo]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (342 mg, 34.1% yield) was purified by column chromatography.

[1261] 1 H NMR (400MHz, CHLOROFORM-d) δ = 10.10-9.91 (m, 1H), 5.06 (br dd, J1 = 3.2, J2 = 6.2Hz, 2H), 4.26 (ddd, J1 = 3.2, J2 = 5.8, J3 = 11.8Hz, 2H), 4.11 (br s,1H),4.01(td,J1=5.8,J2=11.6Hz,2H),3.43(t,J=6.3Hz,5H),3.21-2.87(m,6H),2.35(br t,J=7.4Hz,8H),1.99(br d,J=11.8Hz,10H),1.78-1.66(m,8H),1.65-1.58(m,10H),1.54-1.38(m,12H),1.26(s,32H),1.05-0.76(m,18H)ppm.

[1262] LCMS:RT=2.591,m / z 1053.5[M+H] + .

[1263] Preparation Example 50: Preparation of Compound 47

[1264] Step 1: Synthesis of 1-(benzyloxy)decane-3-ol

[1265] Reaction formula:

[1266] Material proportions:

[1267] Operation process:

[1268] 3-(benzyloxy)propanal (2.0 g, 12.1 mmol) was dissolved in tetrahydrofuran (20 mL), and magnesium bromo(heptyl)magnesium (2.73 g, 1.0 M, 13.4 mL, 13.4 mmol) was added at 0 °C. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. After the reaction was completed by TLC, 50 mL of ammonium chloride aqueous solution was added, and the mixture was extracted twice with 50 mL of ethyl acetate each time. The organic phase was concentrated and purified by column chromatography to give a colorless liquid 1-(benzyloxy)decane-3-ol (2.05 g, 63.6% yield).

[1269] Step 2: Synthesis of 1-(benzyloxy)decane-3-ylhexyl ester

[1270] Reaction formula:

[1271] Material proportions:

[1272] Operation process:

[1273] 1-(benzyloxy)decane-3-ol (2.00 g, 7.56 mmol) and hexanoic acid (966 mg, 8.32 mmol) were dissolved in dichloromethane (20 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.18 g, 11.3 mmol) and 4-dimethylaminopyridine (184 mg, 1.51 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots. 10 mL of water was added, and the mixture was extracted twice with 10 mL of dichloromethane each time. The organic phase was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-(benzyloxy)decane-3-ylhexyl ester (2.00 g, 72.9% yield).

[1274] Step 3: Synthesis of 1-hydroxydecane-3-ylhexyl ester

[1275] Reaction formula:

[1276] Material proportions:

[1277] Operation process:

[1278] 1-(benzyloxy)decane-3-ylhexyl ester (2.00 g, 5.52 mmol) was dissolved in methanol (20 mL), and palladium on carbon (1.17 g, 1.10 mmol) was added. The mixture was reacted at 35 °C and 40 Psi for 12 hours under hydrogen protection. The reaction was monitored by TLC until completion, and then filtered and concentrated. The colorless liquid 1-hydroxydecane-3-ylhexyl ester (880 mg, 58.5% yield) was purified by column chromatography.

[1279] Step 4: Synthesis of 3-(hexanoyloxy)decyl 8-bromooctyl ester

[1280] Reaction formula:

[1281] Material proportions:

[1282] Operation process:

[1283] 1-Hydroxydecane-3-ylhexyl ester (880 mg, 3.23 mmol) and 8-bromooctanoic acid (792 mg, 3.55 mmol) were dissolved in dichloromethane (10 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (928 mg, 4.85 mmol) and 4-dimethylaminopyridine (78.9 mg, 0.646 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots. 10 mL of water was added, and the mixture was extracted twice with 10 mL of dichloromethane each time. The organic phase was then concentrated. The crude product was purified by column chromatography to give a bright yellow liquid, 3-(hexanoyloxy)decyl 8-bromooctyl ester (1.30 g, 84.2% yield).

[1284] Step 5: Synthesis of 3-(hexanoyloxy)decyl 8-[(8-{[3-(hexanoyloxy)decyl]oxo}-8-oxoylide octyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1285] Reaction formula:

[1286] Material proportions:

[1287] Operation process:

[1288] 3-(hexanoyloxy)decyl 8-bromooctyl ester (1.20 g, 2.51 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (130 mg, 1.13 mmol), potassium carbonate (1.04 g, 7.54 mmol), potassium iodide (625 mg, 3.77 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a bright yellow liquid, 3-(hexanoyloxy)decyl 8-[(8-{[3-(hexanoyloxy)decyl]oxo}-8-oxoylidecyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (680 mg, 29.8% yield).

[1289] 1H NMR(400MHz, CHLOROFORM-d)δ=5.04-4.93(m,2H),4.15-4.04(m,5H),3.37-3.23(m,1H),3.11-2.99(m,2H),2.96-2.84(m,2H),2.29(dt,J1 =2.0, J2=7.6Hz,8H),1.94(s,9H),1.92-1.83(m,8H),1.67-1.61(m,8H),1.57-1.52(m,3H),1.41-1.22(m,41H),0.89(q,J=7.2Hz,12H)ppm.

[1290] LCMS:RT=2.410,m / z 909.3[M+H] + .

[1291] Preparation Example 51: Preparation of Compound 48

[1292] Step 1: Synthesis of 1-(benzyloxy)nonane-2-ol

[1293] Reaction formula:

[1294] Material proportions:

[1295] Operation process:

[1296] 3-(benzyloxy)propanal (4.0 g, 26.6 mmol) was dissolved in tetrahydrofuran (40 mL), and magnesium bromo(heptyl)magnesium (5.96 g, 1.0 M, 29.3 mL, 29.3 mmol) was added at 0 °C. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. After the reaction was completed by TLC, 50 mL of ammonium chloride aqueous solution was added, and the mixture was extracted twice with 50 mL of ethyl acetate each time. The organic phase was concentrated and purified by column chromatography to give a colorless liquid 1-(benzyloxy)nonane-2-ol (5.62 g, 84.2% yield).

[1297] Step 2: Synthesis of 1-(benzyloxy)nonane-2-ylhexyl ester

[1298] Reaction formula:

[1299] Material proportions:

[1300] Operation process:

[1301] 1-(benzyloxy)nonane-2-ol (2.00 g, 7.99 mmol) and n-hexanoic acid (1.02 g, 8.79 mmol) were dissolved in dichloromethane (20 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.30 g, 11.9 mmol) and 4-dimethylaminopyridine (195 mg, 1.60 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots. 10 mL of water was added, and the mixture was extracted twice with 10 mL of dichloromethane each time. The organic phase was then concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-(benzyloxy)nonane-2-ylhexyl ester (2.60 g, 93.3% yield).

[1302] Step 3: Synthesis of 1-hydroxynonane-2-ylhexyl ester

[1303] Reaction formula:

[1304] Material proportions:

[1305] Operation process:

[1306] 1-(benzyloxy)nonane-2-ylhexyl ester (2.60 g, 7.46 mmol) was dissolved in methanol (30 mL), and palladium on carbon (1.59 g, 1.49 mmol) was added. The mixture was reacted at 35 °C and 40 Psi for 12 h under hydrogen protection. The reaction was monitored by TLC until completion, and then filtered and concentrated. The colorless liquid 1-hydroxynonane-2-ylhexyl ester (1.50 g, 77.8% yield) was purified by column chromatography.

[1307] Step 4: Synthesis of 2-(hexanoyloxy)nonyl-8-bromooctyl ester

[1308] Reaction formula:

[1309] Material proportions:

[1310] Operation process:

[1311] 1-Hydroxynonane-2-ylhexyl ester (1.50 g, 5.81 mmol) and 8-bromooctanoic acid (1.55 g, 6.97 mmol) were dissolved in dichloromethane (15 mL). Then, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.67 g, 8.71 mmol) and 4-dimethylaminopyridine (141 mg, 1.16 mmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. TLC monitoring showed the formation of new spots. 10 mL of water was added, and the mixture was extracted twice with 10 mL of dichloromethane each time. The organic phase was then concentrated. The crude product was purified by column chromatography to give a bright yellow liquid, 2-(hexanoyloxy)nonyl-8-bromooctyl ester (2.43 g, 90.3% yield).

[1312] Step 5: Synthesis of 2-(hexanoyloxy)nonyl8-[(8-{[2-(hexanoyloxy)nonyl]oxo}-8-oxoylide octyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1313] Reaction formula:

[1314] Material proportions:

[1315] Operation process:

[1316] 2-(hexanoyloxy)nonyl 8-bromooctyl ester (1.20 g, 2.59 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (134 mg, 1.17 mmol), potassium carbonate (1.07 g, 7.77 mmol), potassium iodide (644 mg, 3.88 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the mixture was filtered and concentrated. The solution was purified by column chromatography to obtain a bright yellow liquid, 2-(hexanoyloxy)nonyl 8-[(8-{[2-(hexanoyloxy)nonyl]oxo}-8-oxoylidecyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (750 mg, 32.8% yield).

[1317] 1H NMR (400MHz, CHLOROFORM-d) δ = 5.09 (dq, J1 = 3.6, J2 = 6.8Hz, 2H), 4.23 (dd, J1 = 3.2, J2 = 12.0Hz, 2H), 4.10 (br s,1H),4.06-3.99(m,2H),3.35-3.23(m,1H),3.11-2.98(m,2H),2.97-2 .83(m,2H),2.31(dt,J1=2.0,J2=7.6Hz,8H),2.10-2.03(m,2H),1.90(br d,J=4.0Hz,8H),1.88-1.84(m,3H),1.68-1.64(m,3H),1.61-1.54(m,9H),1.39-1.22(m,40H),0.97-0.79(m,12H)ppm.

[1318] LCMS:RT=2.328,m / z 881.3[M+H] + .

[1319] Preparation Example 52: Preparation of Compound 49

[1320] Step 1: Preparation of 49-1

[1321] Reaction formula:

[1322] Material proportions:

[1323] Operation process:

[1324] 8-Bromooctanoic acid, 1,2-epoxyoctane, ferric chloride, and pyridine were added to a reaction flask, and the reaction was carried out at room temperature for 16 h. The product had an Rf value of 0.4 and a TLC (PE:EA = 4:1). The reaction solution was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and then washed 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 obtain 7 g of oil.

[1325] Step 2: Preparation of 49-2

[1326] Reaction formula:

[1327] Material proportions:

[1328] Operation process:

[1329] 49-1, n-valeric acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5℃ to 0℃ for 16 h. The product Rf value was 0.6, and the TLC (PE:EA = 10:1) was measured. The reaction solution was washed once with 150 mL of saturated sodium bicarbonate aqueous solution, and then once with 150 mL of saturated sodium chloride aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to obtain 7 g of oil.

[1330] Step 3: Preparation of Compound 49

[1331] Reaction formula:

[1332] Material proportions:

[1333] Operation process:

[1334] 49-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask, and the reaction was carried out at 75 °C for 16 h. The product had an Rf value of 0.6 and a TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After filtration and concentration, the solution was purified by column chromatography to obtain 2.2 g of oil.

[1335] 1 H NMR(400MHz,Chloroform-d)δ5.08(qd,J=6.7,3.2Hz,2H),4.21(dd,J=11.8,3.3Hz,2H),4.02(dd,J=11.8,6.7Hz,3H),2.64(s,3H),2.3 0(td,J=7.5,5.1Hz,8H),1.89(d,J=13.5Hz,2H),1.82–1.48(m,22H),1.32(tq,J=16.4,5.7,3.9Hz,34H),0.89(dt,J=16.2,7.1Hz,12H).

[1336] MS(ES+)m / z): 824.6 (M) + .

[1337] Preparation Example 53: Preparation of Compound 50

[1338] Step 1: Synthesis of 1-[(6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxo]undecane-2-yloctyl ester

[1339] Reaction formula:

[1340] Material proportions:

[1341] Operation process:

[1342] 1-[(6-bromohexanoyl)oxo]undecane-2-yloctyl ester (1.0 g, 2.03 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexane-1-ol (2.34 g, 20.3 mmol) was added. The reaction was carried out at 40 °C for 24 hours under nitrogen protection. The reaction was monitored by TLC until it was complete. The solution was concentrated and purified by column chromatography to obtain a yellow liquid 1-[(6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxo]undecane-2-yloctyl ester (870 mg, 81.3% yield).

[1343] Step 2: Synthesis of 2-(hexanoyloxy)nonyl8-[(6-{[2-(octanoyloxy)undecyl]oxo}-6-oxoylide)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1344] Reaction formula:

[1345] Material proportions:

[1346] Operation process:

[1347] 2-(hexanoyloxy)nonyl 8-bromooctyl ester (746 mg, 1.61 mmol) was dissolved in acetonitrile (10 mL), and 1-[(6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxo]undecane-2-yloctyl ester (770 mg, 1.46 mmol), potassium carbonate (708 mg, 5.13 mmol), potassium iodide (291 mg, 1.76 mmol), and tetrahydrofuran (3 mL) were added sequentially. The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, then filtered and concentrated. The solution was purified by column chromatography to obtain a bright yellow liquid, 2-(hexanoyloxy)nonyl 8-[(6-{[2-(octanoyloxy)undecyl]oxo}-6-oxylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (850 mg, 63.8% yield).

[1348] 1H NMR(400MHz,CHLOROFORM-d)δ=5.15-5.01(m,2H),4.28-4.21(m,2H),4.10(br s,1H),4.06-3.96(m,2H),3.35-3.22(m,1H),3.04(dt,J1=5.2,J2=12.0Hz,2H),2.96-2.83( m,2H),2.39-2.26(m,8H),2.10-1.89(m,10H),1.88-1.82(m,3H),1.72-1.61(m,8H),1.57(br s,3H),1.44-1.21(m,45H),0.98-0.81(m,12H)ppm.

[1349] LCMS:RT=2.456,m / z 908.8[M+H] + .

[1350] Preparation Example 54: Preparation of Compound 51

[1351] Step 1: Synthesis of 4-[3-(benzyloxy)propoxy]butane-1-ol

[1352] Reaction formula:

[1353] Material proportions:

[1354] Operation process:

[1355] [(3-bromopropoxy)methyl]benzene (3.00 g, 13.0 mmol) and 1,4-butanediol (5.90 g, 65.4 mmol) were dissolved in dimethyl sulfoxide (30.0 mL), and potassium hydroxide (5.51 g, 98.2 mmol) was added. The mixture was then reacted at 25 °C for 1 hour under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was washed and extracted with water. The crude product was then subjected to column chromatography to obtain a colorless liquid, 4-[3-(benzyloxy)propoxy]butane-1-ol (1.97 g, 63.1% yield).

[1356] Step 2: Synthesis of 4-[3-(benzyloxy)propoxy]butyric acid

[1357] Reaction formula:

[1358] Material proportions:

[1359] Operation process:

[1360] 4-[3-(benzyloxy)propoxy]butane-1-ol (1.00 g, 4.20 mmol) was dissolved in acetonitrile (10.0 mL) and water (5.00 mL). Diacetoxyiodobenzene (2.97 g, 9.23 mmol) was added, followed by 2,2,6,6-tetramethylpiperidine oxide (131 mg, 839 μmol). The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. The reaction was monitored by TLC until completion. The reaction solution was extracted with water and ethyl acetate and concentrated. Column chromatography yielded a colorless liquid 4-[3-(benzyloxy)propoxy]butyric acid (918 mg, 86.7% yield).

[1361] Step 3: Synthesis of 1-(benzyloxy)dodecane-2-yl5-(pentane-3-oxy)pentyl ester

[1362] Reaction formula:

[1363] Material proportions:

[1364] Operation process:

[1365] 1-Hydroxydecane-2-ylheptyl ester (324 mg, 1.13 mmol) and 4-[3-(benzyloxy)propoxy]butyric acid (285 mg, 1.13 mmol) were dissolved in dichloromethane (5.00 mL), and then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (325 mg, 1.70 mmol) and 4-dimethylaminopyridine (13.8 mg, 113 μmol) were added sequentially. The reaction was carried out at 25 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to give a colorless liquid, 1-(benzyloxy)dodecane-2-yl5-(pentane-3-oxy)pentyl ester (531 mg, 90.1% yield).

[1366] Step 4: Synthesis of 1-{[4-(3-hydroxypropoxy)butyryl]oxo}decane-2-ylheptyl ester

[1367] Reaction formula:

[1368] Material proportions:

[1369] Operation process:

[1370] 1-(benzyloxy)dodecane-2-yl5-(pentane-3-oxy)pentyl ester (531 mg, 1.02 mmol) was dissolved in methanol (6.00 mL), and palladium / carbon (217 mg, 203 μmol) was added under an argon atmosphere. The reaction was then carried out at 35 °C for 12 hours under a hydrogen (35 Psi) atmosphere. The reaction was monitored by TLC until completion. The reaction solution was filtered and concentrated. Radiation drying yielded a colorless liquid, 1-{[4-(3-hydroxypropoxy)butyryl]oxo}decane-2-ylheptyl ester (439 mg, 99.9% yield).

[1371] Step 5: Synthesis of 1-{[4-(3-bromopropoxy)butyryl]oxo}decane-2-ylheptyl ester

[1372] Reaction formula:

[1373] Material proportions:

[1374] Operation process:

[1375] 1-{[4-(3-hydroxypropoxy)butyryl]oxo}decane-2-ylheptyl ester (389 mg, 903 μmol) was dissolved in dichloromethane (10.0 mL), and triphenylphosphine (473 mg, 1.81 mmol) and carbon tetrabromide (599 mg, 1.81 mmol) were added at 0 °C. The reaction was carried out at 25 °C for 12 hours under a nitrogen atmosphere. The reaction was monitored by TLC until completion. The solution was concentrated and purified by column chromatography to obtain a colorless liquid 1-{[4-(3-bromopropoxy)butyryl]oxo}decane-2-ylheptyl ester (400 mg, 89.7% yield).

[1376] Step Six: Synthesis of 1-{[4-(3-{[3-(4-{[2-(heptanyloxo)decyl]oxo}-4-oxylidenebutoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)butyryl]oxo}decane-2-ylheptyl ester

[1377] Reaction formula:

[1378] Material proportions:

[1379] Operation process:

[1380] 1-{[4-(3-bromopropoxy)butyryl]oxo}decane-2-ylheptyl ester (300 mg, 607 μmol) was dissolved in acetonitrile (4.00 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (33.6 mg, 291 μmol), potassium carbonate (294 mg, 2.13 mmol), potassium iodide (121 mg, 729 μmol), and tetrahydrofuran (3.00 mL). The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, then filtered and concentrated. Purified by column chromatography, the yellow liquid 1-{[4-(3-{[3-(4-{[2-(heptanyloxy)decyl]oxo}-4-oxylidenebutoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)butyryl]oxo}decane-2-ylheptyl ester (163 mg, 27.8% yield) was obtained.

[1381] 1 H NMR (400MHz, CHLOROFORM-d) δ = 9.73-9.56 (m, 1H), 5.09 (tdd, J1 = 3.4, J2 = 6.7, J3 = 10.0Hz, 2H), 4.22 (td, J1 = 3.2, J2 = 11.7Hz, 2H), 4.12 (br d,J=1.8Hz,1H),4.08-3.99(m,2H),3.61-3.49(m,4H),3.48-3.43(m,4H),3.37(br d,J=5.2Hz,2H),3.19-3.01(m,2H),2.43-2.28(m,10H),2.27-1.96(m,9H),1.94-1.81(m,4H),1.68-1.58(m,10H),1.28(br d,J=14.2Hz,36H),1.05-0.76(m,12H)ppm.

[1382] LCMS:RT=2.439,m / z 941.4[M+H] + .

[1383] Preparation Example 55 Preparation of Compound 52

[1384] Step 1: Synthesis of 1-{[2-(3-bromopropoxy)acetyl]oxo}undecane-2-yloctyl ester

[1385] Reaction formula:

[1386] Material proportions:

[1387] Operation process:

[1388] 1-Hydroxyundecane-2-yloctyl ester (450 mg, 1.43 mmol) was dissolved in dichloromethane (5.00 mL), followed by the sequential addition of 2-(3-bromopropoxy)acetic acid (281 mg, 1.43 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (411 mg, 2.15 mmol), and 4-dimethylaminopyridine (17.4 mg, 143 μmol). The reaction was carried out under nitrogen protection at 25 °C for 12 hours. The reaction was monitored by TLC until completion, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid, 1-{[2-(3-bromopropoxy)acetyl]oxo}undecane-2-yloctyl ester (675 mg, 95.5% yield).

[1389] Step 2: Synthesis of 1-{[2-(3-{[3-(2-{[2-(octanoyloxo)undecyl]oxo}-2-oxylideneethoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)acetyl]oxo}undecyl-2-yloctyl ester

[1390] Reaction formula:

[1391] Material proportions:

[1392] Operation process:

[1393] 1-{[2-(3-bromopropoxy)acetyl]oxo}undecane-2-yloctyl ester (575 mg, 1.17 mmol) was dissolved in acetonitrile (6.00 mL), followed by the addition of (1s,4s)-4-aminocyclohexane-1-ol (64.4 mg, 559 μmol), potassium carbonate (563 mg, 4.08 mmol), potassium iodide (232 mg, 1.40 mmol), and tetrahydrofuran (3.00 mL). The reaction was carried out at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until it ended, then filtered and concentrated. Purified by column chromatography, the yellow liquid 1-{[2-(3-{[3-(2-{[2-(octanoyloxo)undecyl]oxo}-2-oxylideneethoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)acetyl]oxo}undecyl-2-yloctyl ester (157 mg, 14.3% yield) was obtained.

[1394] 1H NMR (400MHz, CHLOROFORM-d) δ=9.57-9.24(m,1H),5.18-5.02(m,2H),4.32(dd,J1=3.0,J2=11.6Hz,2H),4.19-3.99(m,7H),3.70(br s,4H),3.55-3.22(m,4H),2.45-2.26(m,8H),2.17-1.90(m,6H),1.73-1.61(m,7H),1.39-1.19(m,48H),0.91-0.79(m,12H)ppm.

[1395] LCMS:RT=2.564,m / z 941.3[M+H] + .

[1396] Example 1: Preparation and Detection of Lipid Nanoparticles (LNPs)

[1397] To verify whether the lipid nanoparticle (LNP) formulation prepared from the ionizable lipid compound disclosed in this application can effectively encapsulate mRNA and maintain the structural integrity of the mRNA, the prepared ionizable lipid compound, distearate phosphatidylcholine (DSPC, purchased from Nippon Seika Co., Ltd., catalog number: S01005), cholesterol (purchased from Nippon Seika Co., Ltd., catalog number: O01001), and dimyristoylglycerol-polyethylene glycol 2000 (DMG-PEG2000, purchased from Guobang Pharmaceutical Co., Ltd., catalog number: O02005) were dissolved in ethanol (manufacturer: Nanjing Chemical Reagent Co., Ltd., purity 99.6%), 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. Using a microfluidic device, the flow rate was controlled at 12 mL / min, and the volume ratio of the mixed lipid ethanol solution to the mRNA solution obtained in the previous step was controlled at 1:3. Lipid nanoparticles were prepared according to an NPK ratio of ionizable lipids to mRNA of 3–15:1. Ethanol was removed by dialyzing in 20 mM Tris acetic acid for 12–24 hours. Finally, the LNP solution was filtered through a sterile filter with a pore size of 0.22 μm (Millex, catalog number: SLGPR33RB) and concentrated by ultrafiltration (Amicon-Ultra, molecular weight cutoff: 10 kDa) to obtain the LNP formulation obtained by encapsulating Fluc mRNA with the ionizable lipids described in this application, DSPC, cholesterol, and DMG-PEG2000. The particle size and polydispersity index (PDI) of each LNP formulation were determined using dynamic light scattering with a Malvern Zetasizer Ultra instrument (manufacturer: Malvern); the encapsulation efficiency of LNPs was determined using the Quant-it Ribogreen RNA quantification kit (manufacturer: ThermoFisher Scientific, catalog number: R11490).

[1398] Table 1

[1399] In this field, a PDI less than 0.3 indicates that the nanoparticles in the LNP formulation are relatively uniform in size; encapsulation efficiency is used to indicate whether the LNP can effectively encapsulate mRNA, and an encapsulation efficiency greater than 70% indicates that the LNP can effectively encapsulate mRNA. The LNPs prepared by this application have a particle size of 60-80 nm, a PDI less than 0.2, and an encapsulation efficiency greater than 90%.

[1400] Example 2: In vivo animal studies of LNP formulation

[1401] In this embodiment, the LNP prepared in Example 1 was injected into 6- to 8-week-old female Balb / C mice (Vitalliwa) via tail vein injection or hind limb intramuscular injection at a dose of 5 μg / mouse (n=3, i.e., 3 mice were injected and tested in each group, and the data presented are the mean values ​​of each group). D-luciferin potassium salt was injected intraperitoneally at specific time points after administration (4 hours, 24 hours, and 48 hours in this embodiment). The luminescence was then detected using an IVIS Spectrum small animal in vivo imaging system (manufacturer: PerkinElmer). The total luminescence intensity of the expression sites (e.g., liver, hind limb administration sites) was statistically analyzed. Higher luminescence intensity indicates higher luciferase expression, meaning better expression of the corresponding LNP preparation in the mouse. This total luminescence intensity was measured using bioluminescence imaging, specifically the luminescence intensity data of the luminescent sites 6 to 15 minutes after intraperitoneal injection of D-luciferin potassium salt. The total luminescence intensity of the in vivo expression area was statistically analyzed using Living Image software (manufacturer: PerkinElmer). Typically, the total luminescence intensity reading of untreated mice measured by in vivo imaging is on the order of 10. 5 .

[1402] Following the in vivo mouse experimental method described above, the LNP formulation from Example 1 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity in the liver region was recorded, and the results are shown in Table 2 and Figure 1. The LNP formulation tested in this example showed strong expression in mice, with AUC ranging from 10-1. 9 ~10 12 This indicates that the LNP formulations corresponding to the ionizable lipids described in the preparation examples can effectively deliver mRNA into the body and express it.

[1403] Table 2. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1404] Example 3: Preparation and Detection of Lipid Nanoparticles (LNPs)

[1405] In this example, compounds 4 and 14 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios shown in Table 3, following the method described in Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using a 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were measured. As shown in Table 3, the LNP formulations prepared in this example had particle sizes between 70 and 90 nm, PDIs less than 0.25, and encapsulation efficiencies greater than 90%.

[1406] Table 3

[1407] Example 4: In vivo animal studies of LNP formulation

[1408] Following the in vivo mouse test method described in Example 2, the LNP reagent prepared in Example 3 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was then measured, and the results are shown in Figure 2 and Table 4. It can be seen that the LNP preparations prepared in this example exhibited strong expression in mice.

[1409] Table 4. Area under the liver region expression kinetics (AUC) from 4 to 48 hours.

[1410] Example 5: Preparation and Detection of Lipid Nanoparticles (LNPs)

[1411] In this example, compounds 6, 7, 10, 12, 13, 17, 19, 21, and 22 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios in Table 5, following the method described in Example 1. The mRNA in this example was diluted in 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were measured. As shown in Table 5, the LNP formulations prepared in this example had particle sizes between 55 and 100 nm, PDIs less than 0.2, and encapsulation efficiencies greater than 85% (except for compound 6).

[1412] Table 5

[1413] Example 6: In vivo animal studies of LNP formulation

[1414] Following the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 5 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in vivo was measured, and the test results are shown in Figure 3 and Table 6. It can be seen that the LNP preparation prepared in this example showed strong expression in mice.

[1415] Table 6. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1416] Example 7: Preparation and Detection of Lipid Nanoparticles (LNPs)

[1417] In this example, compounds 9, 20, 25, 26, 27, 28, and 30 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios in Table 7, following the method described in Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using a 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 7, the LNP formulations prepared in this example had particle sizes between 50 and 80 nm, PDIs less than 0.15, and encapsulation efficiencies greater than 90%.

[1418] Table 7

[1419] Example 8: In vivo animal studies of LNP formulation

[1420] Following the in vivo mouse test method described in Example 2, the LNP reagent prepared in Example 7 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was then measured, and the results are shown in Figure 4 and Table 8. It can be seen that the LNP preparation prepared in this example exhibits strong expression in mice.

[1421] Table 8. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1422] Example 9: Preparation and Detection of Lipid Nanoparticles (LNPs)

[1423] In this example, compounds 8, 15, 23, 24, and 29 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios in Table 9, following the method described in Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using a 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 9, the LNP formulations prepared in this example had particle sizes between 50 and 90 nm, PDIs less than 0.15, and encapsulation efficiencies greater than 85%.

[1424] Compound I-6-II is referenced from international application WO2024017250A1 and Chinese application CN117417264A, and was synthesized using the same preparation method as WO2024017250A1.

[1425] Table 9

[1426] Example 10: In vivo animal studies of LNP formulation

[1427] Following the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 9 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was measured, and the test results are shown in Figure 5 and Table 10. It can be seen that the LNP preparation prepared in this example showed strong expression in mice.

[1428] Table 10. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1429] Example 11 Preparation and Detection of Lipid Nanoparticles (LNPs)

[1430] In this example, compounds 11, 16, 18, 31, and 32 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios in Table 11, following the method described in Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using a 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 11, the LNP formulations prepared in this example had particle sizes between 55 and 120 nm, PDIs less than 0.2, and encapsulation efficiencies higher than 85%.

[1431] Table 11

[1432] Example 12 Animal in vivo study of LNP formulation

[1433] Following the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 11 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was measured, and the test results are shown in Figure 6 and Table 12. It can be seen that the LNP preparation prepared in this example showed strong expression in mice.

[1434] Table 12. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1435] Example 13 Preparation and Detection of Lipid Nanoparticles (LNPs)

[1436] In this example, compounds 53 and 54 were selected as ionizable lipids, and LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N-P ratios in Table 13, following the method described in Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using a 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 13, the LNP formulations prepared in this example had particle sizes between 130 and 140 nm, PDIs less than 0.55, and encapsulation efficiencies greater than 85%.

[1437] Table 13

[1438] Example 14: In vivo animal studies of LNP formulation

[1439] Following the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 13 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was measured, and the test results are shown in Figure 7 and Table 14. It can be seen that the LNP preparation prepared in this example showed strong expression in mice.

[1440] Table 14. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1441] Example 15 Preparation and Detection of Lipid Nanoparticles (LNPs)

[1442] In this example, compounds 33, 34, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, and 46 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios in Table 15, following the method described in Example 1. The mRNA in this example was diluted in 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 15, the LNP formulations prepared in this example had particle sizes between 50 and 90 nm, PDIs less than 0.15, and encapsulation efficiencies greater than 85%.

[1443] Table 15

[1444] Example 16: In vivo animal studies of LNP formulation

[1445] Following the in vivo mouse test method described in Example 2, the LNP reagent prepared in Example 16 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was then measured, and the results are shown in Figure 8 and Table 16. It can be seen that the LNP preparation prepared in this example exhibits strong expression in mice.

[1446] Table 16. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1447] Example 17 Preparation and Detection of Lipid Nanoparticles (LNPs)

[1448] In this example, compounds 35, 47, 48, 49, 50, 51, and 52 were selected as ionizable lipids. LNP formulations (encapsulated with Fluc mRNA) were prepared according to the molar ratios and N / P ratios in Table 17, following the method described in Example 1. The mRNA in this example was diluted in 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 17, the LNP formulations prepared in this example had particle sizes between 45 and 100 nm, PDIs less than 0.25, and encapsulation efficiencies greater than 90%.

[1449] Table 17

[1450] Example 18: In vivo animal studies of LNP formulation

[1451] Following the in vivo mouse test method of Example 2, the LNP reagent prepared in Example 17 was injected intravenously into 6- to 8-week-old female Balb / C mice at a dose of 5 μg / mouse. The total luminescence intensity of the liver tissue in the mice was measured, and the test results are shown in Figure 9 and Table 18. It can be seen that the LNP preparation prepared in this example showed strong expression in mice.

[1452] Table 18. Area under the curve (AUC) of liver region expression kinetics from 4 to 48 hours.

[1453] Example 19 Preparation and Detection of Lipid Nanoparticles (LNPs)

[1454] In this example, compounds 1, 5, 7, 10, 17, and 21 were selected as ionizable lipids. LNP formulations (encapsulated with RSV mRNA, Uniprot:accession No. P03420) were prepared according to the molar ratios and N / P ratios in Table 19, following the method described in Example 1. The mRNA in this example was diluted in 25 mM sodium acetate solution at pH 5.0, and dialysis was performed using 20 mM Tris-acetic acid solution at pH 7.5. The particle size, PDI, and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 19, the LNP formulations prepared in this example had particle sizes between 60 and 90 nm, PDIs less than 0.15, and encapsulation efficiencies greater than 90%.

[1455] Table 19

[1456] Example 20: In vivo animal studies of LNP formulation

[1457] Following the in vivo mouse assay method described in Example 2, the RSV-LNP reagent prepared in Example 19 was injected via tail vein into 6- to 8-week-old female BABL / c mice at a dose of 40 μg / mouse. Whole blood was collected from the mice 6 hours after injection. Serum was separated from the whole blood by centrifugation at 2000g for 10 min at 4°C and stored at -80°C for analysis. The concentration of RSV antigen expression in mouse serum was quantitatively detected by enzyme-linked immunosorbent assay (ELISA) using the Novizan Respiratory Syncytial Virus pre-F Elisa Kit (Φ&Ⅳ) (catalog number: DD3939) according to the manufacturer's instructions. The test results are shown in Figure 10 and Table 20 (serum RSV antigen concentration (n=5), statistically analyzed by ANOVA, ****p<0.0001, *****p<0.0001, compared with the placebo group). Compared with the placebo group, the RSV antigen concentrations in the groups containing compounds 1, 5, 7, 10, 17, and 21 were significantly increased, and significantly better than those in compound I-6-II.

[1458] Table 20. RSV antigen content in serum of BABL / c mice 6 hours after intravenous administration.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in, R 1 A hydroxyl group, or a C substituted with one or more hydroxyl groups. 1-6 alkyl; Ring A is C 3-8 Cycloalkylene; L 1 For chemical bonds or C 1-3 Alkylene; X and Y are independently C 1-15 Alkylene; Z 1 for -O-、-OZ 1c -C(=O)-O- or chemical bond; Z 1a Z 1b and Z 1c Independently for C 1-6 Alkylene; Z 2 for -O-or-OZ 2c -C(=O)-O-; Z 2a Z 2b and Z 2c Independently for C 1-6 Alkylene; W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene; R 2 C 1-15 Alkyl, -R 2a -OC 1-15 alkyl, R 2a For chemical bonds or C 1-6 Alkylene; R 2c and R 2d Independently for C 1-15 Alkyl or C 1-15 alkenyl; R 2b C 1-15 Alkyl, C 1-15 alkenyl, -C 1-15 Alkylene-R 2ba -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 Alkyl; R 2ba It is a 3-10 membered heterocyclic alkyl group; wherein the heterocyclic alkyl group has 1-3 heteroatoms, and the heteroatoms are independently N, O or S; R 3 For H, C 1-15 Alkyl, -R 3a -OC 1-15 alkyl, R 3a For chemical bonds or C 1-6 Alkylene; R 3b C 1-6 Alkylene; R 3c R 3d and R 3e Independently for C 1-15 alkyl; R 4 C 1-15 Alkyl, -R 4a -OC 1-15 alkyl, R 4a For chemical bonds or C 1-6 Alkylene; R 4b C 1-6 Alkylene; R 4c and R 4e Independently for C 1-15 Alkyl or C 1-15 alkenyl; R 4d C 1-15 Alkyl, C 1-15 alkenyl, -C 1-15 Alkylene-R 4da -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 alkyl; R 4da It is a 3-10 membered heterocyclic alkyl group; wherein the heterocyclic alkyl group has 1-3 heteroatoms, and the heteroatoms are independently N, O or S; R 5 For H, C 1-15 alkyl, R 5a C 1-6 Alkylene; R 5b and R 5c Independently for C 1-15 alkyl.

2. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, Z 1 for Or chemical bonds; Z 1a and Z 1b Independently for C 1-6 Alkylene; Z 2 for Z 2a and Z 2b Independently for C 1-6 Alkylene; R 2a For chemical bonds or C 1-6 Alkylene; R 2b R 2c and R 2d Independently for C 1-15 Alkyl or C 1-15 alkenyl; R 4c R 4d and R 4e Independently for C 1-15 Alkyl or C 1-15 Alkenyl group.

3. The compound of formula I as claimed in claim 2, or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1)R 1 C is a hydroxyl group or a C substituted with a hydroxyl group. 1-3 alkyl; (2) Ring A is C 4-6 Cycloalkylene; (3)L 1 It is a chemical bond or a methylene group; (4) X and Y are independently C 3-10 Alkylene; (5)Z 1a and Z 1b Independently for C 1-3 Alkylene; (6)Z 1 for Or a chemical bond, * indicates the end connected to X; (7)Z 2a and Z 2b Independently for C 1-4 Alkylene; (8)Z 2 for * indicates the end connected to Y; (9)W 1 and W 2 Independent of chemical bonds or C 1-3 Alkyl groups, such as chemical bonds, methylene groups, (10)R 2a For chemical bonds or C 1-3 Alkylene; (11)R 2b C 5-15 Alkyl or C 5-15 alkenyl; (12)R 2c C 5-15 alkyl; (13)R 2d C 1-15 alkyl; (14)R 2 C 5-15 Alkyl, -R 2a -OC 5-15 alkyl, R 2 Preferred is -R 2a -OC 1-15 alkyl, (15)R 3a C 1-3 Alkylene; (16)R 3b C 1-3 Alkylene; (17)R 3c C 5-15 alkyl; (18)R 3d C 5-15 alkyl; (19)R 3e C 1-15 alkyl; (20)R 3 For H, C 5-15 Alkyl, -R 3a -OC 5-15 alkyl, (21)R 4a It is a chemical bond; (22)R 4b C 1-3 Alkylene; (23)R 4c C 5-15 alkyl; (24)R 4d C 5-15 Alkyl or C 5-15 alkenyl; (25)R 4e C 5-15 alkyl; (26)R 4 C 5-15 Alkyl, -R 4a -OC 5-15 alkyl, (27)R 5a C 1-3 Alkylene; (28)R 5b C 5-15 alkyl; (29)R 5c C 5-15 alkyl; (30)R 5 For H, C 5-15 alkyl, 4. The compound of formula I as claimed in claim 3, or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1)R 1 for Preferred (2) Ring A is (3) for Preferably, for More preferably for (4)L 1 It is a chemical bond; (5) X and Y are independent (6)Z 1a and Z 1b for (7)Z 1 for Or a chemical bond, * indicates the end connected to X; (8)Z 2a and Z 2b Independently (9)Z 2 for * indicates the end connected to Y; (10)W 1 and W 2 Independently for C 1-3 Alkylenes, such as methylene, (11)R 2a It is a chemical bond or a methylene group; (12)R 2b for (13)R 2c for (14)R 2d for (15)R 2 for (16)R 3a It is methylene; (17)R 3b It is methylene; (18)R 3c for (19)R 3d for (20)R 3e for (21)R 3 For H, (22) for (23)R 4b It is methylene; (24)R 4c for (25)R 4d for (26)R 4e for (27)R 4 for (28)R 5a It is methylene; (29)R 5b for (30)R 5c for (31)R 5 For H, (32) for 5. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1) X and Y are independent (2)Z 1a and Z 1b Independently methylene or (3)Z 1c C 1-3 Alkylenes, such as methylene, (4)Z 1 for -O- or * -OZ 1c -C(=O)-O-, * indicates the end connected to X; Preferably, Z 1 for -O-、 * indicates the end connected to X; More preferably, Z 1 for -O-、 * indicates the end connected to X; (5)Z 2a and Z 2b Independently (6)Z 2c C 1-4 Alkylenes, such as methylene, (7)Z 2 for -O- or * -OZ 2c -C(=O)-O-, * indicates the end connected to Y; Preferably, Z 2 for -O-、 * indicates the end connected to Y; More preferably, Z 2 for -O-、 * indicates the end connected to Y; (8)R 2b -C 3-15 Alkylene-R 2ba -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 alkyl; Preferably, R 2b for (9)R 2ba It is a 5-8 membered heterocyclic alkyl group; preferably, the heterocyclic alkyl group has 2 heteroatoms, and the heteroatoms are preferably S; Preferably, R 2ba for (10)R 2 for (11)R 3e C 5-15 Alkyl, for example (12)R 3 for (13) for (14)R 4d -C 3-15 Alkylene-R 4da -C 1-6 Alkylene-OC 3-8 Alkyl or -C 1-6 Alkylene-SC 3-8 alkyl; Preferably, R 4d for (15)R 4da It is a 5-8 membered heterocyclic alkyl group; preferably, the heterocyclic alkyl group has 2 heteroatoms, and the heteroatoms are preferably S; Preferably, R 4da for (16)R 4 for (17)R 5c for (18)R 5 for (19) for (20) In the compound of formula I, R 2c R 2d R 2b R 4c R 4e and R 4d In, the C 1-15 The alkenyl group is independently C 2-15 Alkenyl group.

6. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound of formula I satisfies any of the following conditions: The compound of formula I described in scheme (1) has the following structure: Among them, R 1 Ring A, L 1 X, Y, Z 1 Z 2 W 1 W 2 R 2b R 3 R 4 and R 5 The definition is as described in any one of claims 1-5; Preferably, the compound of formula I has the following structure: Among them, R 1 Ring A, L 1 X, Y, Z 1 Z 2 W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any one of claims 1-5; in, Preferred W 1 and W 2 C is preferred 1-3 Alkylene; More preferably, the compound of formula I has the following structure: Among them, R 1 Rings A, X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any one of claims 1-5; Among them, ring A is preferred. More preferably, the compound of formula I has the following structure: Among them, X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any one of claims 1-5; The compound of formula I described in scheme (2) has the following structure: Among them, R 1 X, Y, Z 1 Z 2 W 1 W 2 R 2 R 3 R 4 and R 5 The definition is as described in any one of claims 1-5; Preferably, R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl; X and Y are independently C 3-10 Alkylene; Z 1 for * indicates the end connected to X; Z 1a C 1-6 Alkylene; Z 2 for * indicates the end connected to Y; Z 2a Independently for C 1-6 Alkylene; W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene; R 2 C 1-15 alkyl or R 2b C 1-15 alkyl; R 3 C 1-15 alkyl; R 4 C 1-15 alkyl or R 4d C 1-15 alkyl; R 5 C 1-15 alkyl; The compound of formula I described in scheme (3) has the following structure: Among them, R 1 X, Y, W 1 W 2 R 2 R 3 R 4 and R 5 The definition is as described in any one of claims 1-5; Preferably, R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl; X and Y are independently C 3-10 Alkylene; W 1 and W 2 Independently for C 1-6 Alkylene; R 2 C 1-15 alkyl or R 2b C 1-15 alkyl; R 3 C 1-15 alkyl; R 4 C 1-15 alkyl or R 4d C 1-15 alkyl; R 5 C 1-15 alkyl; The compound of formula I described in scheme (4) has the following structure: Among them, R 1 X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any one of claims 1-5; Preferably, R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl; X and Y are independently C 3-10 Alkylene; W 1 and W 2 Independently for C 1-6 Alkylene; R 2b C 1-15 alkyl; R 3 C 1-15 alkyl; R 4d C 1-15 alkyl; R 5 C 1-15 alkyl; More preferably, R 1 It is a hydroxyl group, or -CH2-OH; X and Y are independently C 4-8 Alkylene, preferably C 5-7 Alkylene, for example W 1 and W 2 Independently for C 1-3 Alkyl groups, such as methylene groups; R 2b C 6-10 Alkyl group, preferably C 6-8 Alkyl, for example R 3 C 1-15 Alkyl group, preferably C 8-12 Alkyl, more preferably C 8-10 Alkyl, for example R 4d C 1-15 Alkyl group, preferably C 4-9 Alkyl, more preferably C 5-8 Alkyl, for example R 5 C 1-15 Alkyl group, preferably C 6-12 Alkyl, more preferably C 7-10 Alkyl, for example The compound of formula I described in scheme (5) has the following structure: Among them, X, Y, W 1 W 2 R 2b R 3 R 4d and R 5 The definition is as described in any one of claims 1-5; Preferably, X and Y are independently C 5-7 Alkylene, for example W 1 and W 2 Independently for C 1-3 Alkyl groups, such as methylene groups; R 2b C 6-8 Alkyl, for example R 3 C 8-10 Alkyl, for example R 4d C 5-8 Alkyl, for example R 5 C 7-10 Alkyl, for example The compound of formula I described in scheme (6) has any of the following structures: Among them, R 1 X, Y, Z 1 Z 2 W 1 W 2 R 2 R 3 R 4 and R 5 The definition is as described in any one of claims 1-5; Preferably, the structure of formula I-3A is as shown in I-3Aa: Preferably, the structure of formula I-3B is as shown in I-3Ba: Preferably, in formula I-3A, R 1 A hydroxyl group, or a C substituted with a hydroxyl group. 1-3 alkyl; X and Y are independently C 3-10 Alkylene; Z 1 for * indicates the end connected to X; Z 1a C 1-6 Alkylene; Z 2 for * indicates the end connected to Y; Z 2a C 1-6 Alkylene; W 1 and W 2 Independent of chemical bonds or C 1-6 Alkylene; R 2 C 1-15 alkyl or R 2b C 1-15 alkyl; R 3 C 1-15 alkyl; R 4 C 1-15 alkyl or R 4d C 1-15 alkyl; R 5 C 1-15 alkyl; Preferably, in formula I-3B, R 1 It is a hydroxyl group; X and Y are independently C 3-10 Alkylene; Z 1 for Z 1a C 1-6 Alkylene; Z 2 for Z 2a C 1-6 Alkylene; W 1 and W 2 It is a chemical bond; R 2 C 1-15 alkyl; R 3 C 1-15 alkyl; R 4 C 1-15 alkyl; R 5 C 1-15 alkyl; Preferably, in formula I-3C, R 1 It is a hydroxyl group; X and Y are independently C 3-10 Alkylene; Z 1 for * indicates the end connected to X; Z 2 for * indicates the end connected to Y; W 1 and W 2 Independently for C 1-6 Alkylene; R 2 for R 2b C 1-15 alkyl; R 3 C 1-15 alkyl; R 4 for R 4d C 1-15 alkyl; R 5 C 1-15 alkyl.

7. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound of formula I is any of the following compounds:

8. A lipid carrier comprising substance Z, said substance Z being a compound of formula I as described in any one of claims 1-7 or a pharmaceutically acceptable salt thereof.

9. The lipid carrier as described in claim 8, characterized in that, It meets one or more of the following conditions: (1) The lipid carrier also includes a diluent, such as phosphate buffer or Tris buffer; (2) The lipid carrier also includes phospholipids, which are phospholipid molecules with polar ends and nonpolar ends of fatty chains, such as distearylphosphatidylcholine, myristoylphosphatidylcholine, dioleoylphosphatidylcholine, palmitoylphosphatidylcholine, 1,2-distearylphosphatidylcholine, docosylphosphatidylcholine or palmitoylphosphatidylcholine. (3) The lipid carrier also includes PEG lipids, which are lipid molecules modified with the hydrophilic end of polyethylene glycol, such as PEG-modified myristoyl glycerol; (4) The lipid carrier further includes sterols, which are selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassosterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol; (5) In the lipid carrier, the molar ratio of substance Z to 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 substance Z to phospholipid is 1-15:1, preferably 2-8:1, for example 3-6:1; (7) In the lipid carrier, the molar ratio of substance Z to phospholipid is 1-15:1, preferably 1-10:1, for example 5:1; (8) In the lipid carrier, the molar ratio of substance Z to PEG lipid is 10-100:1, preferably 10-50:1, for example 33.3:1; (9) In the lipid carrier, the molar content of 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) The lipid carrier is composed of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol; (14) The formulation of the lipid carrier is as follows: Substance Z: Phospholipid: Sterol: PEG lipid is 50:10:38.5:1.5; The preferred substance Z is... The phospholipid is preferably DSPC; the sterol is preferably cholesterol; and the PEG lipid is preferably DMG-PEG2000.

10. The use of a compound of formula I as described in any one of claims 1-7 or a pharmaceutically acceptable salt thereof, or a lipid carrier as described in any one of claims 8-9, in the preparation of a nucleic acid prophylactic agent and / or therapeutic agent delivery carrier; The nucleic acid therapeutic and / or preventive agents are 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, and 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.

11. A lipid nanoparticle comprising a nucleic acid preventive agent and / or therapeutic agent and a lipid carrier as described in any one of claims 8-9; The nucleic acid therapeutic and / or preventive agents are 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, and 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. The lipid nanoparticles according to claim 11, 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, and even more preferably (3-16):1, for example 6:1; (2) The lipid nanoparticles have a particle size of 10-200 nm, preferably 40-150 nm, and more preferably 50-80 nm, such as 71.19 nm, 59.77 nm, 60.70 nm, 62.58 nm or 65.27 nm. (3) The polydispersity index of the lipid nanoparticles is 0.001-0.15, for example 0.036, 0.043, 0.068, 0.072 or 0.101; (4) The encapsulation efficiency of the lipid nanoparticles is 90%-100%, for example 94.4%, 95.7%, 96.8%, 95.8% or 96.1%; (5) In the lipid nanoparticles, the lipid carrier encapsulates the nucleic acid preventive agent and / or therapeutic agent; (6) The particle size (average particle size) of the lipid nanoparticles is 40-150 nm, for example 51.06 nm, 54.04 nm, 56.72 nm, 57.13 nm, 58.09 nm, 61.61 nm, 62.63 nm, 64.82 nm, 66.13 nm, 68.31 nm, 70.58 nm, 71.20 nm, 77.44 nm, 77.93 nm, 80.03 nm, 84.49 nm, 87.13 nm, 87.86 nm, 89.65 nm, 98.29 nm or 118.03 nm; (7) The polydispersity index of the lipid nanoparticles is 0.001-0.30, for example 0.023, 0.038, 0.041, 0.046, 0.053, 0.053, 0.056, 0.058, 0.060, 0.069, 0.087, 0.088, 0.090, 0.091, 0.092, 0.096, 0.097, 0.103, 0.105, 0.108, 0.122, 0.124, 0.125, 0.130, 0.144, 0.179, 0.206 or 0.224; (8) The encapsulation efficiency of the lipid nanoparticles is 85%-100%, for example 87.3%, 89.0%, 89.1%, 89.4%, 90.9%, 92.1%, 92.1%, 92.2%, 92.4%, 93.4%, 94.1%, 95.2%, 95.3%, 95.5%, 95.7%, 95.7%, 96.0%, 96.2%, 96.2%, 96.2%, 96.6%, 96.7%, 96.8%, 96.9%, 96.9%, 97.1%, 97.7%, or 97.9%.

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