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

Abstract

The invention discloses a nitrogen-containing chain compound, a preparation method, a composition containing the nitrogen-containing chain compound and application. The invention specifically discloses a compound shown as a formula I or pharmaceutically acceptable salt thereof. The compound has the following advantages that a prepared LNP preparation is relatively uniform in nanoparticle size, relatively high in encapsulation efficiency and high in in-vivo expression activity.

Description

Technical Field

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

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

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

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

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

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

[0007]

[0008] Among them, R 1 is a hydroxyl group, or a C substituted by 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)-

[0013] O- or chemical bonds;

[0014] Z 1a 、Z 1b and Z 1c Independently C 1-6 alkylene;

[0015] Z 2 for -O- or -OZ 2c -C(=O)-O-;

[0016] Z 2a 、Z 2b and Z 2c Independently C 1-6 alkylene;

[0017] W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0018] R 2 C 1-15 Alkyl, -R 2a -OC 1-15 alkyl,

[0019] R 2a For chemical bonds or C 1-6 Alkylene; R 2c and R 2d Independently C 1-15 Alkyl or C 1-15 alkenyl;

[0020] 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 is a 3-10 membered heterocycloalkyl group; the number of heteroatoms in the heterocycloalkyl group is 1-3, and the heteroatoms are independently N, O or S;

[0021] R 3 H, C 1-15 Alkyl, -R 3a -OC 1-15 alkyl,

[0022] R 3a For chemical bonds or C 1-6 Alkylene; R 3b C 1-6 Alkylene; R 3c 、R 3d and R 3e Independently C 1-15 alkyl;

[0023] R 4 C 1-15 Alkyl, -R 4a -OC 1-15 alkyl,

[0024] R 4a For chemical bonds or C 1-6 Alkylene; R 4b C 1-6 alkylene;

[0025] R 4c and R 4e Independently C 1-15 Alkyl or C 1-15 alkenyl;

[0026] 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;

[0027] R 4da is a 3-10 membered heterocycloalkyl group; the number of heteroatoms in the heterocycloalkyl group is 1-3, and the heteroatoms are independently N, O or S;

[0028] R 5 H, C 1-15 alkyl,

[0029] R 5a C 1-6 Alkylene; R 5b and R 5c Independently C 1-15 alkyl.

[0030] The present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof,

[0031]

[0032] Among them, R 1 is a hydroxyl group, or a C substituted by one or more hydroxyl groups 1-6 alkyl;

[0033] Ring A is C 3-8 cycloalkylene;

[0034] L 1 For chemical bonds or C 1-3 alkylene;

[0035] X and Y are independently C 1-15 alkylene;

[0036] Z 1 for or chemical bond; Z 1a and Z 1b Independently C 1-6 alkylene;

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

[0038] W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0039] R 2 C 1-15 Alkyl, -R 2a -OC 1-15 alkyl, R 2a For chemical bonds or C 1-6 Alkylene; R 2b 、R 2c and R 2d Independently C 1-15 Alkyl or C 1-15 alkenyl;

[0040] R 3 H, C 1-15 Alkyl, -R 3a -OC 1-15 alkyl, R 3a For chemical bonds or C 1-6 Alkylene; R3b C 1-6 Alkylene; R 3c 、R 3d and R 3e Independently C 1-15 alkyl;

[0041] 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;

[0042] R 4c 、R 4d and R 4e Independently C 1-15 Alkyl or C 1-15 alkenyl;

[0043] R 5 H, C 1-15 alkyl, R 5a C 1-6 Alkylene; R 5b and R 5c Independently C 1-15 alkyl.

[0044] In certain preferred embodiments of the present invention, certain groups of the compound of Formula I or a pharmaceutically acceptable salt thereof are defined as follows. Unmentioned groups are the same as those described in any embodiment of the present invention (referred to as "in a certain embodiment of the present invention"). For other inventions of the present invention, the same abbreviations as the present invention are used and are not further described.

[0045] In one embodiment of the present invention, R 1 is a hydroxyl group or a C substituted by a hydroxyl group 1-3 alkyl.

[0046] In one embodiment of the present invention, R 1 for

[0047] In one embodiment of the present invention, R 1 for

[0048] In one embodiment of the present invention, ring A is C 4-6 Cycloalkylene, e.g.

[0049] In one embodiment of the present invention, for

[0050] Preferably, for

[0051] In one embodiment of the present invention, for

[0052] In one embodiment of the present invention, L 1 It is a chemical bond or a methylene group, preferably a chemical bond.

[0053] In one embodiment of the present invention, X and Y are independently C 3-10 Alkylene.

[0054] In one embodiment of the present invention, X and Y are independently

[0055] In one embodiment of the present invention, X and Y are independently

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

[0057] In one embodiment of the present invention, Z 1a and Z 1b Independently C 1-3 Alkylene, e.g.

[0058] In one embodiment of the present invention, Z 1a and Z 1b Independently C 1-3 Alkylene, such as methylene or

[0059] In one embodiment of the present invention, Z 1c C 1-3 Alkylene, such as methylene,

[0060] In one embodiment of the present invention, Z 1 for or chemical bond, * indicates the end connected to X.

[0061] In one embodiment of the present invention, Z 1 for -O- or *-OZ 1c -C(=O)-O-, * indicates the end connected to X

[0062] In one embodiment of the present invention, Z 1 for -O-, * -OZ 1c -C(=O)-O- or a chemical bond, * represents the end connected to X.

[0063] In one embodiment of the present invention, Z 1 for or chemical bonds;

[0064] Preferably, Z 1 for or chemical bond, * indicates the end connected to X.

[0065] In one embodiment of the present invention, Z 1 for -O-, * indicates the end connected to X;

[0066] Preferably, Z 1 for -O-, * indicates the end connected to X.

[0067] In one embodiment of the present invention, Z 1 for -O-, or chemical bonds;

[0068] Preferably, Z 1 for -O-, or chemical bond, * indicates the end connected to X.

[0069] In one embodiment of the present invention, Z 2a and Z 2b Independently C 1-4 Alkylene, e.g.

[0070] In one embodiment of the present invention, Z 2a and Z 2b Independently C 1-4 Alkylene, e.g.

[0071] In one embodiment of the present invention, Z 2c C 1-4 Alkylene, such as methylene,

[0072] In one embodiment of the present invention, Z2 for *Indicates the end connected to Y.

[0073] In one embodiment of the present invention, Z 2 for -O- or *-OZ 2c -C(=O)-O-, * represents the end connected to Y.

[0074] In one embodiment of the present invention, Z 2 for -O- or *-OZ 2c -C(=O)-O-, * represents the end connected to Y.

[0075] In one embodiment of the present invention, Z 2 for

[0076] Preferably, Z 2 for * indicates the end connected to Y.

[0077] In one embodiment of the present invention, Z 2 for -O-, * indicates the end connected to Y;

[0078] Preferably, Z 2 for -O-, * indicates the end connected to Y.

[0079] In one embodiment of the present invention, Z 2 for -O-,

[0080] Preferably, Z 2 for -O-, *Indicates the end connected to Y.

[0081] In one embodiment of the present invention, W 1 and W 2 Independently C 1-6 Alkylene, preferably C 1-3 Alkylene.

[0082] In one embodiment of the present invention, W 1 and W 2 independently a chemical bond or C 1-3 Alkylene.

[0083] In one embodiment of the present invention, W 1 and W 2 are independently chemical bonds, methylene groups,

[0084] In one embodiment of the present invention, R 2a For chemical bonds or C 1-3 Alkylene.

[0085] In one embodiment of the present invention, R 2a It is a chemical bond or a methylene group.

[0086] In a certain embodiment 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 Alkenyl is independently C 2-15 Alkenyl.

[0087] In a certain embodiment of the present invention, in the compound of formula I, each "C 1-15 "Alkenyl" is independently "C 2-15 "Alkenyl".

[0088] In one embodiment of the present invention, R 2b C 5-15 Alkyl or C 5-15 Alkenyl.

[0089] In one embodiment of the present invention, 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.

[0090] In one embodiment 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.

[0091] In one embodiment of the present invention, R 2ba is a 5-8 membered heterocycloalkyl group; in the heterocycloalkyl group, the number of heteroatoms is preferably 2, and the heteroatom is preferably S.

[0092] In one embodiment of the present invention, R 2ba for

[0093] In one embodiment of the present invention, R 2b for

[0094] In one embodiment of the present invention, R 2b for

[0095] In one embodiment of the present invention, R 2b for

[0096] In one embodiment of the present invention, R 2c C 5-15 alkyl.

[0097] In one embodiment of the present invention, R 2c for

[0098] In one embodiment of the present invention, R 2d C 1-15 alkyl.

[0099] In one embodiment of the present invention, R 2d for In one embodiment of the present invention, R 2 C 5-15 Alkyl, -R 2a -OC 5-15 alkyl, R 2 Preferably -R 2a -OC 1-15 alkyl, In one embodiment of the present invention, R 2 for

[0100] In one embodiment of the present invention, R 2 for

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

[0102]

[0103] In one embodiment of the present invention, R3a C 1-3 Alkylene groups, such as methylene.

[0104] In one embodiment of the present invention, R 3b C 1-3 Alkylene groups, such as methylene.

[0105] In one embodiment of the present invention, R 3c C 5-15 Alkyl groups, e.g. In one embodiment of the present invention, R 3d C 5-15 Alkyl groups, e.g. In one embodiment of the present invention, R 3e C 1-15 Alkyl groups, e.g. In one embodiment of the present invention, R 3e C 5-15 Alkyl groups, e.g. In one embodiment of the present invention, R 3e C 5-15 Alkyl groups, e.g.

[0106] In one embodiment of the present invention, R 3 H, C 5-15 Alkyl, -R 3a -OC 5-15 alkyl,

[0107] In one embodiment of the present invention, R 3 For H,

[0108] In one embodiment of the present invention, R 3 for

[0109] In one embodiment of the present invention, R 3 For H,

[0110] In one embodiment of the present invention, for

[0111] In one embodiment of the present invention, for

[0112] In one embodiment of the present invention, for

[0113]

[0114] In one embodiment of the present invention, R 4a For chemical bonds.

[0115] In one embodiment of the present invention, R 4b C 1-3 Alkylene groups, such as methylene.

[0116] In one embodiment of the present invention, R 4c C 5-15 Alkyl groups, e.g.

[0117] In one embodiment of the present invention, R 4d C 5-15 Alkyl or C 5-15 Alkenyl, e.g.

[0118] In one embodiment of the present invention, 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.

[0119] In one embodiment 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.

[0120] In one embodiment of the present invention, R 4da is a 5-8 membered heterocycloalkyl group; in the heterocycloalkyl group, the number of heteroatoms is preferably 2, and the heteroatom is preferably S.

[0121] In one embodiment of the present invention, R 4da for

[0122] In one embodiment of the present invention, R 4d for

[0123] In one embodiment of the present invention, R 4d for

[0124] In one embodiment of the present invention, R 4e C 5-15 Alkyl groups, e.g.

[0125] In one embodiment of the present invention, R 4 C 5-15 Alkyl, -R 4a -OC 5-15 alkyl,

[0126] In one embodiment of the present invention, R 4 for

[0127] In one embodiment of the present invention, R 4 for

[0128] In one embodiment of the present invention, R 4 for

[0129] In one embodiment of the present invention, R 5a C 1-3 Alkylene groups, such as methylene.

[0130] In one embodiment of the present invention, R 5b C 5-15 Alkyl groups, e.g. In one embodiment of the present invention, R 5c C 5-15 Alkyl groups, e.g. In one embodiment of the present invention, R 5c C 5-15 Alkyl groups, e.g. In one embodiment of the present invention, R 5c C 5-15 Alkyl groups, e.g.

[0131] In one embodiment of the present invention, R 5 H, C 5-15 alkyl, In one embodiment of the present invention, R 5 For H,

[0132] In one embodiment of the present invention, R 5 for

[0133] In one embodiment of the present invention, R 5 For H,

[0134] In one embodiment of the present invention, for

[0135] In one embodiment of the present invention, for

[0136]

[0137]

[0138] In one embodiment of the present invention, for

[0139]

[0140] In one embodiment of the present invention, the compound of formula I has the following structure:

[0141]

[0142] 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; preferably, the compound of formula I has the following structure:

[0143]

[0144] 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 5is defined as in any of the preceding schemes; wherein, Preferably W 1 and W 2 Preferably C 1-3 alkylene;

[0145] More preferably, the compound of formula I has the following structure:

[0146]

[0147] Among them, R 1 , Ring A, X, Y, W 1 、W 2 、R 2b 、R 3 、R 4d and R 5 The definition of is as described in the previous scheme;

[0148] Among them, ring A is preferably

[0149] Further preferably, the compound of formula I has the following structure:

[0150]

[0151] Among them, X, Y, W 1 、W 2 、R 2b 、R 3 、R 4d and R 5 The definition of is as described in the previous scheme.

[0152] In one embodiment of the present invention, the compound of formula I has the following structure:

[0153]

[0154] 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; preferably, R 1 is a hydroxyl group, or a C substituted by a hydroxyl group 1-3 alkyl;

[0155] X and Y are independently C 3-10 alkylene;

[0156] Z 1 for * indicates the end connected to X; Z 1a C 1-6 alkylene;

[0157] Z 2 for * indicates the end connected to Y; Z 2a Independently C 1-6 Alkylene; W 1 and W 2 independently a chemical bond or C 1-6 alkylene;

[0158] R 2 C 1-15 Alkyl or R 2b C 1-15 alkyl;

[0159] R 3 C 1-15 alkyl;

[0160] R 4 C 1-15 Alkyl or R 4d C 1-15 alkyl;

[0161] R 5 C 1-15 alkyl.

[0162] In one embodiment of the present invention, the compound of formula I has the following structure:

[0163]

[0164] Among them, R 1 , X, Y, W 1 、W 2 、R 2 、R 3 、R 4 and R 5 The definition of is as described in the previous scheme;

[0165] Preferably, R 1 is a hydroxyl group, or a C substituted by a hydroxyl group 1-3 alkyl;

[0166] X and Y are independently C 3-10 alkylene;

[0167] W 1 and W 2 Independently C 1-6 alkylene;

[0168] R 2 C 1-15 Alkyl or R 2b C 1-15 alkyl;

[0169] R 3 C 1-15 alkyl;

[0170] R 4 C 1-15 Alkyl or R 4d C 1-15 alkyl;

[0171] R 5 C 1-15 alkyl.

[0172] In one embodiment of the present invention, the compound of formula I has the following structure:

[0173]

[0174] Among them, R 1 , X, Y, W 1 、W 2 、R 2b 、R 3 、R 4d and R 5 The definition of is as described in the previous scheme;

[0175] Preferably,

[0176] R 1 is a hydroxyl group, or a C substituted by a hydroxyl group 1-3 alkyl;

[0177] X and Y are independently C 3-10 alkylene;

[0178] W 1 and W 2 Independently C 1-6 alkylene;

[0179] R 2b C 1-15 alkyl;

[0180] R 3 C 1-15 alkyl;

[0181] R 4d C 1-15 alkyl;

[0182] R 5 C1-15 alkyl;

[0183] More preferably,

[0184] R 1 is hydroxyl, or -CH2-OH;

[0185] X and Y are independently C 4-8 Alkylene, preferably C 5-7 Alkylene, e.g.

[0186] W 1 and W 2 Independently C 1-3 alkylene groups, such as methylene;

[0187] R 2b C 6-10 Alkyl, preferably C 6-8 Alkyl groups, e.g.

[0188] R 3 C 1-15 Alkyl, preferably C 8-12 Alkyl, more preferably C 8-10 Alkyl groups, e.g.

[0189] R 4d C 1-15 Alkyl, preferably C 4-9 Alkyl, more preferably C 5-8 Alkyl groups, e.g.

[0190] R 5 C 1-15 Alkyl, preferably C 6-12 Alkyl, more preferably C 7-10 Alkyl groups, e.g.

[0191] In one embodiment of the present invention, the compound of formula I has the following structure:

[0192]

[0193] Among them, X, Y, W 1 、W 2 、R 2b 、R 3 、R 4d and R 5 The definition of is as described in the previous scheme;

[0194] Preferably,

[0195] X and Y are independently C 5-7 Alkylene, e.g. W 1 and W 2 Independently C 1-3 alkylene groups, such as methylene;

[0196] R 2b C 6-8 Alkyl groups, e.g. R 3 C 8-10 Alkyl groups, e.g.

[0197] R 4d C 5-8 Alkyl groups, e.g.

[0198] R 5 C 7-10 Alkyl groups, e.g.

[0199] In one embodiment of the present invention, the compound of formula I is any of the following structures:

[0200]

[0201] 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; preferably, the structure of formula I-3A is shown as I-3Aa:

[0202]

[0203] Preferably, the structure of formula I-3B is as shown in I-3Ba:

[0204]

[0205] Preferably, in formula I-3A,

[0206] R 1 is a hydroxyl group, or a C substituted by a hydroxyl group 1-3 alkyl;

[0207] X and Y are independently C 3-10 alkylene;

[0208] 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 independently a chemical bond or C 1-6 alkylene;

[0209] R 2 C 1-15 Alkyl or R 2b C 1-15 alkyl;

[0210] R 3 C 1-15 alkyl;

[0211] R 4 C 1-15 Alkyl or R 4d C 1-15 alkyl;

[0212] R 5 C 1-15 alkyl;

[0213] Preferably, in formula I-3B,

[0214] R 1 is hydroxyl group;

[0215] X and Y are independently C 3-10 alkylene;

[0216] Z 1 for Z 1a C 1-6 alkylene;

[0217] Z 2 for Z 2a C 1-6 alkylene;

[0218] W 1 and W 2 is a chemical bond;

[0219] R 2 C1-15 alkyl;

[0220] R 3 C 1-15 alkyl;

[0221] R 4 C 1-15 alkyl;

[0222] R 5 C 1-15 alkyl;

[0223] Preferably, in formula I-3C,

[0224] R 1 is hydroxyl group;

[0225] X and Y are independently C 3-10 alkylene;

[0226] Z 1 for * indicates the end connected to X;

[0227] Z 2 for * indicates the end connected to Y;

[0228] W 1 and W 2 Independently C 1-6 alkylene;

[0229] R 2 for R 2b C 1-15 alkyl;

[0230] R 3 C 1-15 alkyl;

[0231] R 4 for R 4d C 1-15 alkyl;

[0232] R 5 C 1-15 alkyl.

[0233] In one embodiment of the present invention, the compound of formula I is any one of the following compounds:

[0234]

[0235]

[0236]

[0237]

[0238]

[0239] The present invention also provides a use of the compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a nucleic acid preventive agent and / or therapeutic agent delivery vector;

[0240] The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small hairpin RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA) or messenger RNA (mRNA), preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 spike protein (Spike) mRNA, herpes zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0241] The present invention also provides a composition comprising a substance Z, wherein the substance Z is a compound of formula I as described above or a pharmaceutically acceptable salt thereof.

[0242] The present invention also provides a lipid carrier, which includes a substance Z, wherein the substance Z is the compound of formula I as described above or a pharmaceutically acceptable salt thereof.

[0243] In one embodiment of the present invention, the lipid carrier further comprises a diluent, which can be a phosphate buffer or a Tris buffer.

[0244] In one embodiment of the present invention, the lipid carrier further comprises phospholipids.

[0245] In one embodiment of the present invention, the phospholipid may be a conventional phospholipid in the art, which is an amphiphilic auxiliary molecule that helps the fusion of lipid particles and cell membranes. The phospholipid may be a phospholipid molecule having an electrically charged polar end and a non-polar end of a fatty chain, such as distearoylphosphatidylcholine (DSPC), dimyristoylphosphocholine (DMPC), dioleoylphosphocholine (DOPC), palmitoylphosphocholine (DPPC), 1,2-distearoylphosphocholine (DSPC), heneicosanoylphosphocholine (DUPC) or palmitoylphosphocholine (POPC).

[0246] In one embodiment of the present invention, the lipid carrier further comprises PEG lipid (polyethylene glycol-modified lipid).

[0247] In one embodiment of the present invention, the PEG lipid may be a lipid molecule having a polyethylene glycol hydrophilic end modified. The PEG lipid is preferably selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol, such as PEG-modified dimyristoylglycerol (DMG-PEG2000).

[0248] In one embodiment of the present invention, the lipid carrier further comprises sterol.

[0249] In one embodiment of the present invention, the sterol may be a conventional sterol in the art, including animal, plant or fungal sterols. The sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid and α-tocopherol, such as cholesterol.

[0250] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the sterol is 0.5-5:1, preferably 0.5-3:1, for example 1.3:1.

[0251] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 2-8:1, such as 3-6:1.

[0252] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 1-10:1, for example 5:1.

[0253] In one embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to the PEG lipid is 10-100:1, preferably 10-50:1, for example 33.3:1.

[0254] In the present invention, molar content means the percentage of a substance in the total mass of the lipid carrier, and the sum of the molar contents of the components in the lipid carrier does not exceed 100 mol%.

[0255] In one embodiment of the present invention, the molar content of the substance Z in the lipid carrier is 30 mol% to 70 mol%, for example, 50 mol%.

[0256] In one embodiment of the present invention, the molar content of the phospholipid in the lipid carrier is 5 mol% to 20 mol%, for example, 10 mol%.

[0257] In one embodiment of the present invention, the molar content of the sterol in the lipid carrier is 20 mol% to 60 mol%, for example, 38.5 mol%.

[0258] In one embodiment of the present invention, the molar content of the PEG lipid in the lipid carrier is about 0.2 mol% to 5 mol%, for example, 1.5% mol.

[0259] In a certain embodiment of the present invention, in the lipid carrier, the lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.

[0260] In one embodiment of the present invention, the lipid carrier has the following formula: substance Z: phospholipid: sterol: PEG lipid in a ratio of 50:10:38.5:1.5;

[0261] The substance Z is preferably The phospholipid is preferably DSPC; the sterol is preferably cholesterol; and the PEG lipid is preferably DMG-PEG2000.

[0262] The present invention also provides a use of the lipid carrier in preparing a delivery vector for a nucleic acid preventive agent and / or therapeutic agent;

[0263] The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small hairpin RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA) or messenger RNA (mRNA), preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 spike protein (Spike) mRNA, herpes zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0264] The present invention also provides a lipid nanoparticle comprising a nucleic acid preventive agent and / or therapeutic agent and the aforementioned lipid carrier;

[0265] The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid (DNA), double-stranded DNA, small interfering RNA (siRNA), asymmetric double-stranded small interfering RNA (aiRNA), microRNA (miRNA), small hairpin RNA (shRNA), circular RNA (circRNA), transfer RNA (tRNA) or messenger RNA (mRNA), preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 spike protein (Spike) mRNA, herpes zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0266] In one embodiment of the present invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles may be (2-30):1. The nitrogen-to-phosphorus ratio refers to the ratio of the moles of ionizable nitrogen atoms in one or more ionizable lipid compounds to the moles of phosphate groups in the RNA. In this application, it refers to the ratio of the moles of ionizable nitrogen atoms in the ionizable lipid nanoparticles to the moles of phosphate groups in the mRNA in the pharmaceutical composition. Preferably, the nitrogen-to-phosphorus ratio is (2-20):1, preferably (3-20):1, for example (3-16):1.

[0267] In one embodiment of the present invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles is 6:1.

[0268] In one embodiment of the present invention, the particle size (average particle size) of the lipid nanoparticles is 10-200 nm, preferably 40-150 nm, and more preferably 50-80 nm, such as 71.19 nm, 59.77 nm, 60.70 nm, 62.58 nm or 65.27 nm.

[0269] In one embodiment of the present invention, 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.

[0270] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.001-0.15, such as 0.036, 0.043, 0.068, 0.072 or 0.101.

[0271] 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.

[0272] In one embodiment of the present invention, the encapsulation efficiency of the lipid nanoparticles is 90%-100%, such as 94.4%, 95.7%, 96.8%, 95.8% or 96.1%.

[0273] In one embodiment 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%.

[0274] In a certain embodiment of the present invention, in the lipid nanoparticles, the lipid carrier encapsulates the nucleic acid preventive agent and / or therapeutic agent.

[0275] Unless otherwise specified, the terms used in this invention may be defined as follows:

[0276] It will be understood by those skilled in the art that the structural formulas used in the present invention to describe groups are based on the conventions used in the art. It means that the corresponding group is connected to other fragments and groups in the compound through this site.

[0277] As used herein, a substituent may be preceded by a single dash "-" to indicate that the named substituent is bonded to the parent moiety through a single bond.

[0278] When one of the variables is selected from a chemical bond, it means that the two groups it connects are directly connected.

[0279] When any variable appears multiple times in the definition of a compound, the definition of the variable at each position is independent of the definition at the remaining positions, and their meanings are independent of each other and do not affect each other.

[0280] The term "plurality" refers to 2, 3 or 4.

[0281] The term "alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, and the like.

[0282] The term "alkylene" refers to a straight or branched divalent hydrocarbon group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkylene include, but are not limited to, methylene, ethylene, propylene, 1-methylpropylene, butylene, and the like.

[0283] The term "alkenyl" refers to a straight or branched hydrocarbon group having a specified number of carbon atoms (e.g., C2-C4) and one or more double bonds. The one or more carbon-carbon double bonds may be internal or terminal.

[0284] The term "cycloalkylene" refers to a saturated cyclic, divalent group having a specified number of ring carbon atoms (e.g., C3-C8) and the ring atoms consisting only of carbon atoms. Examples of cycloalkyl groups include, but are not limited to wait.

[0285] The term "pharmaceutically acceptable salt" refers to salts prepared from compounds of the present invention with relatively nontoxic, pharmaceutically acceptable acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent.

[0286] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0287] The term "prevention" refers to a period of time in which a subject remains healthy relative to a disease or condition mentioned herein. It should be understood that the period depends on the amount of the drug compound that has been administered and the individual factors of the subject discussed elsewhere in this specification. It should be understood that prevention may not be effective in all subjects treated with the compounds according to the present invention. However, the term requires that a statistically significant portion of a cohort or group of subjects is preferably effectively prevented from suffering from a disease or condition or its associated symptoms as referred to herein. Preferably, in this case, it is envisioned that a group or group of subjects would typically develop a disease or condition as referred to herein without taking preventive measures according to the present invention. Those skilled in the art can use various well-known statistical evaluation tools discussed elsewhere in this specification to immediately determine whether a portion is statistically significant.

[0288] On the basis of conforming to the common sense in this field, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0289] The reagents and raw materials used in the present invention are commercially available.

[0290] The positive progress of the present invention is that the LNP preparation prepared by using the nitrogen-containing chain compound of the present invention has relatively uniform nanoparticle size, high encapsulation efficiency and high in vivo expression activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0291] Figure 1 is the chemiluminescence intensity of the mouse liver area after tail vein administration of compound 1, compound 2, compound 3 and compound 5;

[0292] Figure 2 The chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 4 and compound 14;

[0293] Figure 3 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of Compound 6, Compound 7, Compound 10, Compound 12, Compound 13, Compound 17, Compound 19, Compound 21 and Compound 22;

[0294] Figure 4 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 9, compound 20, compound 25, compound 26, compound 27, compound 28 and compound 30;

[0295] Figure 5 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound I-6-II, compound 8, compound 15, compound 23 and compound 29;

[0296] Figure 6 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 11, compound 16, compound 18, compound 31 and compound 32;

[0297] Figure 7 is the chemiluminescence intensity of the mouse liver area after tail vein administration of compound 53 and compound 54;

[0298] Figure 8 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of Compound 33, Compound 34, Compound 36, Compound 37, Compound 38, Compound 39, Compound 40, Compound 41, Compound 43, Compound 44, Compound 45 and Compound 46;

[0299] Figure 9 is the chemiluminescence intensity of the liver area of ​​mice after tail vein administration of compound 35, compound 47, compound 48, compound 50, compound 51 and compound 52;

[0300] Figure 10The expression of RSV antigen in mouse serum after tail vein administration of Compound I-6-II, Compound 7, Compound 10, Compound 17, Compound 21, Compound 1 and Compound 5 (6 hours after administration, n=5). DETAILED DESCRIPTION

[0301] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0302] Preparation Example 1 Preparation of Compound 1

[0303] Step 1: Preparation of 1-1

[0304] Reaction formula:

[0305]

[0306] Material ratio:

[0307] Material Name Molecular weight Feed ratio Feeding amount mmol Succinic anhydride 100 4eq 35g 350 8-pentadecanol 228 1eq 20g 88 DMAP 122 1eq 11g 88 Triethylamine 101 0.5 eq 4.5g 44 DCM - - 400mL -

[0308] Operation process:

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

[0310] Step 2: Preparation of 1-2

[0311] Reaction formula:

[0312]

[0313] Material ratio:

[0314] Material Name Molecular weight Feed ratio Feeding amount mmol 1-1 328.49 1eq 3.3g 10 6-bromohexanol 139 1 eq 1.4g 10 EDCI 192 1.5eq 3g 15 DMAP 122 0.15 eq 200mg 1.5 DCM - - 80mL -

[0315] Operation process:

[0316] 1-1,6-bromohexanol, EDCI, DMAP, and DCM were added to the reaction flask and allowed to react at room temperature for 16 h. The product had an Rf value of 0.5 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 2.7 g of an oil.

[0317] Step 3: Preparation of compound 1

[0318] Reaction formula:

[0319]

[0320] Material ratio:

[0321] Material Name Molecular weight Feed ratio Feeding amount mmol 1-2 491.55 2.5eq 2.15g 4.4 cis-3-Aminocyclohexanol hydrochloride 115.18 1 eq 200mg 1.74 potassium carbonate 138 3 eq 720mg 5.2 Potassium iodide 166 1 eq 300mg 1.74 Acetonitrile - - 30mL -

[0322] Operation process:

[0323] 1-2, cis-3-aminocyclohexanol hydrochloride, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 65°C for 16 hours. The product had an rR value of 0.6, as determined by TLC (DCM:MeOH = 20:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.1 g of an oil.

[0324] 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).

[0325] MS (ES+) m / z): 936.7 (M) + .

[0326] Preparation Example 2 Preparation of Compound 2

[0327] Step 1: Preparation of 2-1

[0328] Reaction formula:

[0329]

[0330] Material ratio:

[0331] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195.06 1 eq 19.5g 100 1,2-Epoxydodecane 184.32 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mg 2.5

[0332] Operation process:

[0333] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and stirred at room temperature for 16 hours. TLC (PE:EA = 4:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 200 mL of ethyl acetate and washed once with 200 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 27 g of a colorless oil.

[0334] Step 2: Preparation of 2-2

[0335] Reaction formula:

[0336]

[0337] Material ratio:

[0338]

[0339]

[0340] Operation process:

[0341] Add 2-1, octanoic acid, EDCI, DMAP, and DCM to a reaction flask and stir at room temperature for 2 h. TLC (PE:EA = 20:1) reveals an Rf value of 0.6 for the product. Wash the reaction mixture once with 200 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 20 g of a colorless oil.

[0342] Step 3: Preparation of compound 2

[0343] Reaction formula:

[0344]

[0345] Material ratio:

[0346] Material Name Molecular weight Feed ratio Feeding amount mmol 2-2 505.58 2.5 eq 2.23g 4.4 cis-3-aminocyclohexanol 115.18 1 eq 200mg 1.74 potassium carbonate 138 3 eq 720mg 5.2 Potassium iodide 166 1 eq 300mg 1.74 Acetonitrile - - 30mL -

[0347] Operation process:

[0348] Add 2-2-, cis-3-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile to a reaction flask and react at 65°C for 16 hours. TLC (DCM:MeOH = 10:1) reveals an Rf value of 0.5 for the product. Dilute the reaction solution with 100 mL of ethyl acetate and wash once with 100 mL of water. The organic phase is dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1 g of an oil.

[0349] 1 H 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).

[0350] MS (ES+) m / z): 965.8 (M+H) + .

[0351] Preparation Example 3 Preparation of Compound 3

[0352] Step 1: Synthesis of 1-(benzyloxy)dodecan-3-ol

[0353] Reaction formula:

[0354]

[0355] Material ratio:

[0356]

[0357]

[0358] Operation process:

[0359] Dissolve 3-(Benzyloxy)propanal (6.00 g, 36.5 mmol) in tetrahydrofuran (60.0 mL). Slowly add a solution of nonylmagnesium bromide (10.1 g, 43.8 mmol) in tetrahydrofuran at 0°C. Incubate at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. Column chromatography of the crude product affords 1-(benzyloxy)dodecan-3-ol (5.87 g) as a colorless liquid.

[0360] Step 2: Synthesis of 1-(benzyloxy)dodec-3-yloctyl ester

[0361] Reaction formula:

[0362]

[0363] Material ratio:

[0364]

[0365] Operation process:

[0366] Dissolve 1-(Benzyloxy)dodecan-3-ol (5.87 g, 20.0 mmol) and n-octanoic acid (3.47 g, 24.0 mmol) in dichloromethane (58.0 mL). Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.77 g, 30.1 mmol) and 4-dimethylaminopyridine (245 mg, 2.01 mmol) sequentially. The mixture is reacted at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC, and the reaction solution is concentrated. Column chromatography of the crude product affords 1-(Benzyloxy)dodecan-3-yloctyl ester (6.00 g) as a colorless liquid.

[0367] Step 3: Synthesis of 1-hydroxydodecane-3-yl octyl ester

[0368] Reaction formula:

[0369]

[0370] Material ratio:

[0371] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodec-3-yloctyl ester 418 1.0eq 6.00g 14.3 Palladium / Carbon 106 0.20eq 3.05g 2.87 hydrogen - (40Psi) - - Methanol - - 60.0mL -

[0372] Operation process:

[0373] Dissolve 1-(Benzyloxy)dodec-3-yloctyl ester (6.00 g, 14.3 mmol) in methanol (60.0 mL). Add palladium / carbon (3.05 g, 2.87 mmol) under argon. Incubate at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Monitor the reaction by TLC. Filter and concentrate the reaction mixture to obtain a colorless liquid, 1-hydroxydodec-3-yloctyl ester (4.60 g).

[0374] Step 4: Synthesis of 1-[(6-bromohexanoyl)oxy]dodec-3-yloctyl ester

[0375] Reaction formula:

[0376]

[0377] Material ratio:

[0378]

[0379] Operation process:

[0380] Dissolve 1-hydroxydodec-3-yloctyl ester (2.00 g, 6.09 mmol) in dichloromethane (15.0 mL). Add 6-bromohexanoic acid (1.42 g, 7.31 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.75 g, 9.13 mmol), and 4-dimethylaminopyridine (74.3 mg, 608 μmol) in this order. The mixture is reacted at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. The crude product is purified by column chromatography to yield 1-[(6-bromohexanoyl)oxy]dodec-3-yloctyl ester (2.78 g), a colorless liquid.

[0381] Step 5: Synthesis of 1-[(6-{[(1R,3S)-3-hydroxycyclohexyl](6-{[3-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino}hexanoyl)oxy]dodec-3-yloctyl ester

[0382] Reaction formula:

[0383]

[0384] Material ratio:

[0385]

[0386] Operation process:

[0387] 1-[(6-bromohexanoyl)oxy]dodec-3-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (10.0 mL). (1S,3R)-3-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-[(6-{[(1R,3S)-3-hydroxycyclohexyl](6-{[3-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino}hexanoyl)oxy]dodec-3-yloctyl ester (441 mg, 21.6% yield) as a colorless liquid.

[0388] 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.

[0389] LCMS:RT=2.175,m / z 964.8[M+H] + .

[0390] Preparation Example 4 Preparation of Compound 5

[0391] Step 1: Preparation of 5-1

[0392] Reaction formula:

[0393]

[0394] Material ratio:

[0395] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mL 2.5

[0396] Operation process:

[0397] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[0398] Step 2: Preparation of compound 5-2

[0399] Reaction formula:

[0400]

[0401] Material ratio:

[0402] Material Name Molecular weight Feed ratio Feeding amount mmol 5-1 379.38 1 eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0403] Operation process:

[0404] 5-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[0405] Step 3: Preparation of compound 5

[0406] Reaction formula:

[0407]

[0408] Material ratio:

[0409] Material Name Molecular weight Feed ratio Feeding amount mmol 5-2 505.58 2.2eq 4g 7.9 cis-4-aminocyclohexanol 115.17 1 eq 414mg 3.6 potassium carbonate 138 3 eq 1.5g 10.8 Potassium iodide 166 1 eq 600mg 3.6 Acetonitrile - - 50mL -

[0410] Operation process:

[0411] 5-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 65°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 20:1). The reaction solution was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.8 g of an oil.

[0412] 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).

[0413] MS (ES+) m / z): 964.8 (M) + .

[0414] Preparation Example 5 Preparation of Compound 4

[0415] Step 1: Synthesis of 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)[(1R,3S)-3-hydroxycyclohexyl]amino]hexyl ester

[0416] Reaction formula:

[0417]

[0418] Material ratio:

[0419]

[0420]

[0421] Operation process:

[0422] 2-(Hexanoyloxy)dodecyl 6-bromohexyl ester (770 mg, 1.61 mmol) was dissolved in acetonitrile (6.0 mL), and (1S,3R)-3-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(hexanoyloxy)dodecyl 6-[(6-{[2-(hexanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)[(1R,3S)-3-hydroxycyclohexyl]amino]hexyl ester as a colorless liquid (400 mg, 25.8% yield).

[0423] 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).

[0424] LCMS:RT=3.994,m / z 908.7[M+H] + .

[0425] Preparation Example 6 Preparation of Compound 14

[0426] Step 1: Synthesis of 1-(benzyloxy)-3-(octanoyloxy)propan-2-yl octyl ester

[0427] Reaction formula:

[0428]

[0429] Material ratio:

[0430]

[0431] Operation process:

[0432] 2-{[(Hexyloxy)carbonyl]oxy}dodecan-1-ol (1.00 g, 5.49 mmol) and octanoic acid (1.74 g, 12.0 mmol) were dissolved in dichloromethane (10 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.16 g, 16.4 mmol) and 4-dimethylaminopyridine (134 mg, 1.10 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)-3-(octanoyloxy)propan-2-yloctyl ester (2.30 g, 96.4% yield) as a colorless liquid.

[0433] Step 2: Synthesis of 1-hydroxy-3-(octanoyloxy)propan-2-yl octyl ester

[0434] Reaction formula:

[0435]

[0436] Material ratio:

[0437]

[0438] Operation process:

[0439] Dissolve 1-(benzyloxy)-3-(octanoyloxy)propan-2-yl octyl ester (2.30 g, 5.29 mmol) in methanol (25 mL), add palladium on carbon (1.13 g, 1.06 mmol), and react under hydrogen protection at 35°C, 40 Psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1-hydroxy-3-(octanoyloxy)propan-2-yl octyl ester (1.80 g, 98.7% yield) as a colorless liquid.

[0440] Step 3: Synthesis of 1-[(6-bromohexanoyl)oxy]-3-(octanoyloxy)propan-2-yloctyl ester

[0441] Reaction formula:

[0442]

[0443] Material ratio:

[0444]

[0445] Operation process:

[0446] 1-Hydroxy-3-(octanoyloxy)propan-2-yl octyl ester (1.80 g, 5.23 mmol) and 6-bromohexanoic acid (1.22 g, 6.27 mmol) were dissolved in dichloromethane (20 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.50 g, 7.84 mmol) and 4-dimethylaminopyridine (127 mg, 1.05 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-[(6-bromohexanoyl)oxy]-3-(octanoyloxy)propan-2-yl octyl ester (2.60 g, 98.0% yield) as a colorless liquid.

[0447] Step 4: Synthesis of 1-{[6-({6-[2,3-di(octanoyloxy)propoxy]-6-oxyylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexanoyl]oxy}-3-(octanoyloxy)propan-2-yloctyl ester

[0448] Reaction formula:

[0449]

[0450] Material ratio:

[0451]

[0452] Operation process:

[0453] 3-[(6-bromohexanoyl)oxy]-2-(octyloxy)propyl octyl ester (1.00 g, 1.97 mmol) was dissolved in acetonitrile (7 mL), and (1s,4s)-4-aminocyclohexan-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) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-{[6-({6-[2,3-di(octanoyloxy)propoxy]-6-oxyidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexanoyl]oxy}-3-(octanoyloxy)propan-2-yl octyl ester (446 mg, 20.8% yield) as a yellow liquid.

[0454] 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), 2.57-2.3 8(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(brs,37H),0.98-0.80(m,12H)ppm.

[0455] LCMS:RT=2.411,m / z 997.2[M+H] + .

[0456] Preparation Example 7 Preparation of Compound 6

[0457] Step 1: Preparation of 6-1

[0458] Reaction formula:

[0459]

[0460] Material ratio:

[0461] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mL 2.5

[0462] Operation process:

[0463] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[0464] Step 2: Preparation of 6-2

[0465] Reaction formula:

[0466]

[0467] Material ratio:

[0468] Material Name Molecular weight Feed ratio Feeding amount mmol 6-1 379.38 1eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0469] Operation process:

[0470] 6-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated aqueous sodium bicarbonate solution, and once with 100 mL of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[0471] Step 3: Preparation of compound 6

[0472] Reaction formula:

[0473]

[0474] Material ratio:

[0475] Material Name Molecular weight Feed ratio Feeding amount mmol 6-2 505 2.5eq 2.5g 5 2-Aminocyclohexanol 115 1 eq 230mg 2 potassium carbonate 138 3 eq 830mg 6 Potassium iodide 166 1 eq 330mg 2 Acetonitrile - - 30mL -

[0476] Operation process:

[0477] 6-2,2-Aminocyclohexanol, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 75°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.2 g of a colorless oil.

[0478] 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).

[0479] MS (ES+) m / z): 964.0 (M) + .

[0480] Preparation Example 8 Preparation of Compound 7

[0481] Step 1: Preparation of 7-1

[0482] Reaction formula:

[0483]

[0484] Material ratio:

[0485] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mL 2.5

[0486] Operation process:

[0487] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[0488] Step 2: Preparation of 7-2

[0489] Reaction formula:

[0490]

[0491] Material ratio:

[0492] Material Name Molecular weight Feed ratio Feeding amount mmol 7-1 379.38 1 eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0493] Operation process:

[0494] 7-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[0495] Step 3: Preparation of compound 7

[0496] Reaction formula:

[0497]

[0498] Material ratio:

[0499] Material Name Molecular weight Feed ratio Feeding amount mmol 7-2 505 2.2eq 2.2g 4.4 3-Aminocyclopentanol 101 1 eq 200mg 2 potassium carbonate 138 3 eq 840mg 6 Potassium iodide 166 1 eq 330mg 2 Acetonitrile - - 30mL -

[0500] Operation process:

[0501] 7-2,3-Aminocyclopentanol, K2CO3, KI, and acetonitrile were added to a reaction flask and stirred at 75°C for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 800 mg of a colorless oil.

[0502] 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).

[0503] MS (ES+) m / z): 950.0 (M) + .

[0504] Preparation Example 9 Preparation of Compound 10

[0505] Step 1: Preparation of 10-1

[0506] Reaction formula:

[0507]

[0508] Material ratio:

[0509] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mL 2.5

[0510] Operation process:

[0511] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[0512] Step 2: Preparation of 10-2

[0513] Reaction formula:

[0514]

[0515] Material ratio:

[0516] Material Name Molecular weight Feed ratio Feeding amount mmol 10-1 379.38 1 eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0517] Operation process:

[0518] 10-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated aqueous sodium bicarbonate solution, and once with 100 mL of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[0519] Step 3: Preparation of compound LQ210-54

[0520] Reaction formula:

[0521]

[0522] Material ratio:

[0523] Material Name Molecular weight Feed ratio Feeding amount mmol 10-2 505 2.5eq 2g 3.8 cis-4-Aminocyclohexanemethanol hydrochloride 165 1 eq 250mg 1.5 potassium carbonate 138 3 eq 620mg 4.5 Potassium iodide 166 1 eq 250mg 1.5 Acetonitrile - - 30mL -

[0524] Operation process:

[0525] To a reaction flask, 10-2, cis-4-aminocyclohexanemethanol hydrochloride, K2CO3, KI, and acetonitrile were added and stirred at 75°C for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 800 mg of a colorless oil.

[0526] 1 H 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).

[0527] MS (ES+) m / z): 978.0 (M) + .

[0528] Preparation Example 10 Preparation of Compound 12

[0529] Step 1: Preparation of 12-1

[0530] Reaction formula:

[0531]

[0532] Material ratio:

[0533] Material Name Molecular weight Feed ratio Feeding amount mmol Heptadecan-9-ol 256 1eq 7.7g 30 1,6-Hexanedioic acid 146 1.5 eq 6.6g 45 EDCI 192 1.5eq 8.7g 45 DMAP 122 0.15eq 550mg 4.5 DCM - - 200mL -

[0534] Operation process:

[0535] Heptadecane-9-ol, 1,6-hexanedioic acid, EDCI, DMAP, and DCM were added to a reaction flask and allowed to react at room temperature for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.6 for the product. The reaction mixture was washed once with 200 mL of aqueous solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[0536] Step 2: Preparation of 12-2

[0537] Reaction formula:

[0538]

[0539] Material ratio:

[0540] Material Name Molecular weight Feed ratio Feeding amount mmol 12-1 385 1eq 3.45g 9.1 4-Bromobutanol 153 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0541] Operation process:

[0542] 12-1,4-bromobutanol, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product Rf value was 0.6, 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 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 an oil.

[0543] Step 3: Preparation of 12-3

[0544] Reaction formula:

[0545]

[0546] Material ratio:

[0547] Material Name Molecular weight Feed ratio Feeding amount mmol 1-Undecanol 172 1eq 5.2g 30 6-Bromohexanoic acid 195 1.1 eq 6.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0548] Operation process:

[0549] 1-Undecanol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7.5 g of an oil.

[0550] Step 4: Preparation of 12-4

[0551] Reaction formula:

[0552]

[0553] Material ratio:

[0554] Material Name Molecular weight Feed ratio Feeding amount mmol 12-3 349 0.8eq 5.6g 16 cis-4-aminocyclohexanol 115 1 eq 2.3g 20 potassium carbonate 138 1.5 eq 4.2g 30 Acetonitrile - - 80mL -

[0555] Operation process:

[0556] To a reaction flask, 12-3, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 3.5 g of a colorless oil.

[0557] Step 5: Preparation of compound 12

[0558] Reaction formula:

[0559]

[0560] Material ratio:

[0561]

[0562]

[0563] Operation process:

[0564] To a reaction flask, 12-2, 12-4, K2CO3, KI, and acetonitrile were added and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.2 g of a colorless oil.

[0565] 1 H 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).

[0566] MS (ES+) m / z): 822.0 (M) + .

[0567] Preparation Example 11 Preparation of Compound 13

[0568] Step 1: Synthesis of ({[1,3-di(heptyloxy)propan-2-yl]oxy}methyl)benzene

[0569] Reaction formula:

[0570]

[0571] Material ratio:

[0572] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Benzyloxy)propane-1,3-diol 182 1.0 eq 2.00g 10.9 7-Bromoheptan-1-ol 178 2.2eq 4.32g 24.1 Sodium Hydrogen 24 3.0eq 1.32g 32.9 N,N-dimethylformamide - - 20mL - N,N-dimethylformamide - - 10mL -

[0573] Operation process:

[0574] 2-(Benzyloxy)propane-1,3-diol (2.00 g, 10.9 mmol) was dissolved in N,N-dimethylformamide (20 mL). Sodium hydroxide (1.32 g, 32.9 mmol) was added at 25°C and allowed to react 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 mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After addition of 300 mL of water, the mixture was extracted with ethyl acetate twice (100 mL each time). The organic phase was concentrated. Column chromatography of the crude product afforded ({[1,3-di(heptyloxy)propan-2-yl]oxy}methyl)benzene (2.30 g, 55.3% yield) as a yellow liquid.

[0575] Step 2: Synthesis of 1,3-di(heptyloxy)propan-2-ol

[0576] Reaction formula:

[0577]

[0578] Material ratio:

[0579]

[0580]

[0581] Operation process:

[0582] Dissolve 1-(benzyloxy)-3-(octanoyloxy)propan-2-yl octyl ester (2.30 g, 5.29 mmol) in methanol (25 mL), add palladium on carbon (1.13 g, 1.06 mmol), and react under hydrogen at 35°C, 40 psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain a colorless liquid, 1,3-di(heptyloxy)propan-2-ol (1.60 g, 91.3% yield).

[0583] Step 3: Synthesis of 1,3-di(heptyloxy)propan-2-yl 8-bromooctyl ester

[0584] Reaction formula:

[0585]

[0586] Material ratio:

[0587]

[0588] Operation process:

[0589] Dissolve 1,3-di(heptyloxy)propan-2-ol (1.60 g, 5.55 mmol) and 8-bromooctanoic acid (1.48 g, 6.66 mmol) in dichloromethane (20 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.59 g, 8.32 mmol) and 4-dimethylaminopyridine (135 mg, 1.11 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring revealed the formation of new spots. The reaction solution was concentrated. Column chromatography of the crude product afforded 1,3-di(heptyloxy)propan-2-yl 8-bromooctyl ester (2.60 g, 98.0% yield) as a colorless liquid.

[0590] Step 4: Synthesis of undecyl 6-bromohexyl ester

[0591] Reaction formula:

[0592]

[0593] Material ratio:

[0594]

[0595] Operation process:

[0596] Undecan-1-ol (1.77 g, 10.2 mmol) and 6-bromohexanoic acid (2.00 g, 10.2 mmol) were dissolved in dichloromethane (20 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.95 g, 15.3 mmol) and 4-dimethylaminopyridine (250 mg, 2.05 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield undecyl 6-bromohexyl ester (3.37 g, 94.0% yield) as a colorless liquid.

[0597] Step 5: Synthesis of undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester

[0598] Reaction formula:

[0599]

[0600] Material ratio:

[0601] Material Name Molecular weight Feed ratio Feeding amount mmol Undecyl 6-bromohexyl ester 348 1.0 eq 1.30g 3.72 (1s,4s)-4-Aminocyclohexan-1-ol 115 2.0 eq 857mg 7.44 Triethylamine 101 2.0 eq 753mg 7.44 Acetonitrile - - 13mL -

[0602] Operation process:

[0603] Undecyl 6-bromohexyl ester (1.30 g, 3.72 mmol) and (1s,4s)-4-aminocyclohexan-1-ol (857 mg, 7.44 mmol) were dissolved in acetonitrile (13 mL). Triethylamine (753 mg, 7.44 mmol) was added to the reaction mixture. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to afford undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (460 mg, 29.6% yield) as a yellow liquid.

[0604] Step 6: Synthesis of 1,3-di(heptyloxy)propan-2-yl 8-{[6-oxyylidene-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester

[0605] Reaction formula:

[0606]

[0607] Material ratio:

[0608]

[0609] Operation process:

[0610] Undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (400 mg, 1.04 mmol) was dissolved in acetonitrile (4 mL). 1,3-Di(heptyloxy)propan-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 mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1,3-Di(heptyloxy)propan-2-yl 8-{[6-oxyidene-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester (57 mg, 6.30% yield), a colorless liquid.

[0611] 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.

[0612] LCMS:RT=2.628,m / z 796.6[M+H] + .

[0613] Preparation Example 12 Preparation of Compound 17

[0614] Step 1: Preparation of 17-1

[0615] Reaction formula:

[0616]

[0617] Material ratio:

[0618] Material Name Molecular weight Feed ratio Feeding amount mmol Succinic anhydride 100 4eq 8g 80 8-pentadecanol 228 1 eq 4.6g 20 DMAP 122 1 2.5g 20 Triethylamine 101 0.5 1g 10 DCM - - 100mL -

[0619] Operation process:

[0620] Succinic anhydride, 8-pentadecanol, DMAP, triethylamine, and DCM were added to a reaction flask and stirred at room temperature for 16 hours. The product had an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction solution was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 6.5 g of a colorless oil.

[0621] Step 2: Preparation of 17-2

[0622] Reaction formula:

[0623]

[0624] Material ratio:

[0625]

[0626]

[0627] Operation process:

[0628] To the reaction flask, 17-1, 6-bromohexanol, EDCI, DMAP, and DCM were added and allowed to react at room temperature for 16 h. The product had an Rf value of 0.5 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 5.6 g of a colorless oil.

[0629] Step 3: Preparation of compound 17

[0630] Reaction formula:

[0631]

[0632] Material ratio:

[0633] Material Name Molecular weight Feed ratio Feeding amount mmol 17-2 505 2.2eq 2.2g 4.4 cis-4-aminocyclohexanol 115 1 eq 230mg 2 potassium carbonate 138 3 eq 840mg 6 Potassium iodide 166 1 eq 330mg 2 Acetonitrile - - 30mL -

[0634] Operation process:

[0635] To a reaction flask, 17-2, cis-4-aminocyclohexanol, KCO, KI, and acetonitrile were added and heated to 75°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 900 mg of a colorless oil.

[0636] 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).

[0637] MS (ES+) m / z): 936.0 (M) + .

[0638] Preparation Example 13 Preparation of Compound 19

[0639] Step 1: Synthesis of hexyl 4-nitrophenyl carbonate

[0640] Reaction formula:

[0641]

[0642] Material ratio:

[0643] Material Name Molecular weight Feed ratio Feeding amount mmol Hexane-1-ol 130 1.0 eq 2.00g 19.6 4-Nitrophenyl chloride 201 1.2eq 4.73g 23.5 4-Dimethylaminopyridine 122 2eq 4.78g 39.1 dichloromethane - - 10mL -

[0644] Operation process:

[0645] Dissolve hexane-1-ol (2.00 g, 19.6 mmol) in dichloromethane (10 mL). Add 4-methylbenzenesulfonyl chloride (4.78 g, 39.1 mmol) and 4-nitrophenyl chloride (4.73 g, 23.5 mmol) at 0°C. Incubate at 25°C under nitrogen for 2 hours. Monitor the reaction completion by TLC. Wash with water, extract, and concentrate. The crude product, hexyl 4-nitrophenyl carbonate (5.00 g, 95.6% yield), is used directly in the next step.

[0646] Step 2: Synthesis of 1-(benzyloxy)dodec-2-ylhexyl carbonate

[0647] Reaction formula:

[0648]

[0649] Material ratio:

[0650] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecan-2-ol 292 1.0 eq 6.84g 18.7 Hexyl 4-nitrophenyl carbonate 267 1eq 5.00g 18.7 dichloromethane - - 10mL -

[0651] Operation process:

[0652] Dissolve 1-(Benzyloxy)dodecan-2-ol (6.84 g, 18.7 mmol) in dichloromethane (10 mL). Slowly add hexyl 4-nitrophenyl carbonate (5.00 g, 18.7 mmol) at 0°C. The mixture is reacted at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC, and the reaction solution is concentrated. The crude product is purified by column chromatography to afford 1-(benzyloxy)dodecan-2-ylhexyl carbonate (3.50 g, 44.5% yield).

[0653] Step 3: Synthesis of 2-{[(hexyloxy)carbonyl]oxy}dodecan-1-ol

[0654] Reaction formula:

[0655]

[0656] Material ratio:

[0657]

[0658] Operation process:

[0659] 1-(Benzyloxy)dodec-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 mixture was reacted under hydrogen protection at 25°C, 35 psi for 12 hours. The reaction was monitored for completion by TLC, filtered, and concentrated. Purification by column chromatography afforded 2-{[(hexyloxy)carbonyl]oxy}dodecan-1-ol (1.50 g, 100% yield).

[0660] Step 4: Synthesis of 2-{[(hexyloxy)carbonyl]oxy}dodecyl 6-bromohexyl ester

[0661] Reaction formula:

[0662]

[0663] Material ratio:

[0664]

[0665] Operation process:

[0666] 2-{[(Hexyloxy)carbonyl]oxy}dodecan-1-ol (1.50 g, 4.54 mmol) was dissolved in dichloromethane (20.0 mL). 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) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to afford 2-{[(Hexyloxy)carbonyl]oxy}dodecyl 6-bromohexyl ester (1.5 g, 58.6% yield).

[0667] Step 5: Synthesis of 2-{[(hexyloxy)carbonyl]oxy}dodecyl 6-({6-[(2-{[(hexyloxy)carbonyl]oxy}dodecyl)oxy]-6-oxyylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[0668] Reaction formula:

[0669]

[0670] Material ratio:

[0671]

[0672] Operation process:

[0673] 2-{[(Hexyloxy)carbonyl]oxy}dodecyl 6-bromohexyl ester (260 mg, 512 μmol) was dissolved in acetonitrile (1 mL). Ethanolamine (26.6 mg, 231 μmol), potassium carbonate (212 mg, 1.54 mmol), potassium iodide (102 mg, 614 μmol), and tetrahydrofuran (3 mL) were added sequentially. The mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-{[(hexyloxy)carbonyl]oxy}dodecyl 6-({6-[(2-{[(hexyloxy)carbonyl]oxy}dodecyl)oxy]-6-oxyylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (30 mg, 5.93% yield).

[0674] 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.

[0675] LCMS: RT=2.510, m / z=969.3[M+H] + .

[0676] Preparation Example 14 Preparation of Compound 21

[0677] Step 1: Synthesis of heptadecane-9-yl 8-[(5-{[4-(heptanoyloxy)dodecyl]oxy}-5-oxypentylidene)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[0678] Reaction formula:

[0679]

[0680] Material ratio:

[0681]

[0682] Operation process:

[0683] 1-[(5-bromovaleroyl)oxy]dodec-4-ylheptyl ester (1.16 g, 2.42 mmol) was dissolved in acetonitrile (7 mL). Heptadec-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 mixture was reacted at 75°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a colorless liquid, heptadec-9-yl 8-[(5-{[4-(heptanoyloxy)dodecyl]oxy}-5-oxopentyl[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (416 mg, 22.4% yield).

[0684] 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.

[0685] LCMS:RT=2.746,m / z 893.4[M+H] + .

[0686] Preparation Example 15 Preparation of Compound 22

[0687] Step 1: Preparation of 22-1

[0688] Reaction formula:

[0689]

[0690] Material ratio:

[0691] Material Name Molecular weight Feed ratio Feeding amount mmol 7-Bromoheptanoic acid 209 1eq 6.3g 30 1,2-Epoxydecane 156 1 eq 4.7g 30 Ferric chloride 162 0.05 eq 250mg 1.5 Pyridine 79 0.05 eq 120mg 1.5

[0692] Operation process:

[0693] 7-Bromoheptanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 6 g of an oil.

[0694] Step 2: Preparation of 22-2

[0695] Reaction formula:

[0696]

[0697] Material ratio:

[0698] Material Name Molecular weight Feed ratio Feeding amount mmol 22-1 365 1eq 3.3g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0699] Operation process:

[0700] 22-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 3.8 g of an oil.

[0701] Step 3: Preparation of 22-3

[0702] Reaction formula:

[0703]

[0704] Material ratio:

[0705] Material Name Molecular weight Feed ratio Feeding amount mmol 1-nonanol 144 1eq 4.3g 30 6-Bromohexanoic acid 195 1.1 eq 6.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0706] Operation process:

[0707] 1-Nonanol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 6.8 g of an oil.

[0708] Step 4: Preparation of 22-4

[0709] Reaction formula:

[0710]

[0711] Material ratio:

[0712] Material Name Molecular weight Feed ratio Feeding amount mmol 22-3 321 0.8 eq 2.55g 8 cis-4-aminocyclohexanol 115 1eq 1.15g 10 potassium carbonate 138 1.5 eq 2.1g 15 Acetonitrile - - 40mL -

[0713] Operation process:

[0714] 22-3, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.6 g of a colorless oil.

[0715] Step 5: Preparation of compound 22

[0716] Reaction formula:

[0717]

[0718] Material ratio:

[0719] Material Name Molecular weight Feed ratio Feeding amount mmol 22-4 355 1eq 1.6g 4.5 22-2 491 1.2 eq 2.7g 5.4 potassium carbonate 138 1.5 eq 930mg 6.8 Potassium iodide 166 0.5 eq 380mg 2.3 Acetonitrile - - 30mL -

[0720] Operation process:

[0721] 22-4, 22-2, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.6 g of a colorless oil.

[0722] 1 H 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).

[0723] MS (ES+) m / z): 766.0 (M) + .

[0724] Preparation Example 16 Preparation of Compound 9

[0725] Step 1: Preparation of 9-1

[0726] Reaction formula:

[0727]

[0728] Material ratio:

[0729] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mg 2.5

[0730] Operation process:

[0731] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[0732] Step 2: Preparation of 9-2

[0733] Reaction formula:

[0734]

[0735] Material ratio:

[0736] Material Name Molecular weight Feed ratio Feeding amount mmol 9-1 379.38 1eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0737] Operation process:

[0738] 9-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[0739] Step 3: Preparation of compound 9

[0740] Reaction formula:

[0741]

[0742] Material ratio:

[0743] Material Name Molecular weight Feed ratio Feeding amount mmol 9-2 505 2.5eq 2.5g 5 cis-4-(Aminomethyl)cyclohexanol hydrochloride 165.66 1 eq 330mg 2 potassium carbonate 138 3 eq 830mg 6 Potassium iodide 166 1 eq 330mg 2 Acetonitrile - - 30mL -

[0744] Operation process:

[0745] 9-2, cis-4-(aminomethyl)cyclohexanol hydrochloride, K2CO3, KI, and acetonitrile were added to a reaction flask and stirred at 75°C for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.3 g of a colorless oil.

[0746] 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).

[0747] MS (ES+) m / z): 977.9 (M) + .

[0748] Preparation Example 17 Preparation of Compound 20

[0749] Step 1: Synthesis of ethyl (4Z)-oct-4-enyl ester

[0750] Reaction formula:

[0751]

[0752] Material ratio:

[0753]

[0754]

[0755] Operation process:

[0756] (4-Ethoxy-4-oxyylidenebutyl) triphenylphosphane bromide (7.00 g, 18.5 mmol) was dissolved in tetrahydrofuran (70.0 mL), sodium bis(trimethylsilyl)amide (1 M, 19.4 mL) was added at -5°C, and the reaction was carried out at -5°C for 10 mins. n-Butyraldehyde (1.40 g, 19.4 mmol) was added to the reaction solution at -78°C, and the reaction was carried out at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC, quenched, extracted twice with ethyl acetate, and the organic phase was concentrated and purified by column chromatography to obtain colorless liquid ethyl (4Z)-oct-4-enyl ester (450 mg, 14.2% yield).

[0757] Step 2: Synthesis of (4Z)-oct-4-enoic acid

[0758] Reaction formula:

[0759]

[0760] Material ratio:

[0761] Material Name Molecular weight Feed ratio Feeding amount mmol Ethyl (4Z)-oct-4-enyl ester 170 1.00eq 500mg 2.94 Lithium hydroxide monohydrate 42 2.00 eq 246mg 5.87 Tetrahydrofuran - - 5.00mL - Methanol - - 0.50mL - water - - 0.50mL -

[0762] Operation process:

[0763] Ethyl (4Z)-oct-4-enoate (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 under a nitrogen atmosphere for 12 hours. The reaction was monitored by TLC. After acid-base stripping, the mixture was concentrated to give a colorless liquid (4Z)-oct-4-enoic acid (287 mg, 68.7% yield).

[0764] Step 3: Synthesis of 1-[(tert-butyldimethylsilyl)oxy]dodec-2-yl(4Z)-oct-4-enyl ester

[0765] Reaction formula:

[0766]

[0767] Material ratio:

[0768]

[0769] Operation process:

[0770] (2R)-1-[(tert-Butyldimethylsilyl)oxy]dodecan-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 mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave 1-[(tert-Butyldimethylsilyl)oxy]dodecan-2-yl (4Z)-oct-4-enate (537 mg, 60.3% yield) as a colorless liquid.

[0771] Step 4: Synthesis of 1-hydroxydodec-2-yl (4Z)-oct-4-enyl ester

[0772] Reaction formula:

[0773]

[0774] Material ratio:

[0775]

[0776] Operation process:

[0777] 1-[(tert-Butyldimethylsilyl)oxy]dodec-2-yl (4Z)-oct-4-enate (480 mg, 1.09 mmol) was dissolved in tetrahydrofuran (5.00 mL). Triethylamine hydrofluoride (702 mg, 4.36 mmol) was added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. TLC revealed the formation of new spots. After adjusting the pH to neutral, the mixture was extracted with ethyl acetate and the organic phase was concentrated. Purification by column chromatography afforded 1-hydroxydodec-2-yl (4Z)-oct-4-enate (350 mg, 98.4% yield) as a colorless liquid.

[0778] Step 5: Synthesis of 1-[(8-bromooctanoyl)oxy]dodec-2-yl (4Z)-oct-4-enyl ester

[0779] Reaction formula:

[0780]

[0781] Material ratio:

[0782]

[0783] Operation process:

[0784] 1-Hydroxydodec-2-yl (4Z)-oct-4-enate (350 mg, 1.07 mmol) and 8-bromooctanoic acid (334 mg, 1.50 mmol) were dissolved in dichloromethane (5.00 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (308 mg, 1.61 mmol) and 4-dimethylaminopyridine (13.1 mg, 0.107 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave 1-[(8-bromooctanoyl)oxy]dodec-2-yl (4Z)-oct-4-enate (444 mg, 77.9% yield) as a colorless liquid.

[0785] Step 6: Synthesis of 1-[(8-{[8-({2-[(4Z)-oct-4-enoxy]dodecyl}oxy)-8-oxyoctyl][(1s,4s)-4-hydroxycyclohexyl]amino}octanoyl)oxy]dodec-2-yl (4Z)-oct-4-enyl ester

[0786] Reaction formula:

[0787]

[0788] Material ratio:

[0789]

[0790] Operation process:

[0791] Dissolve 1-[(8-bromooctanoyl)oxy]dodec-2-yl (4Z)-oct-4-enate (394 mg, 741 μmol) in acetonitrile (4.00 mL), and add (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) in sequence. Finally, react at 75 ° C under nitrogen protection for 12 hours. The reaction is completed by monitoring by TLC, and the mixture is filtered and concentrated. Purification by column chromatography gave colorless liquid 1-[(8-{[8-({2-[(4Z)-oct-4-enoxy]dodecyl}oxy)-8-oxyoctylidene][(1s,4s)-4-hydroxycyclohexyl]amino}octanoyl)oxy]dodec-2-yl (4Z)-oct-4-enate (202 mg, 23.4% yield).

[0792] 1 H 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.

[0793] LCMS:RT=2.751,m / z 1017.9[M+H] + .

[0794] Preparation Example 18 Preparation of Compound 25

[0795] Step 1: Preparation of 25-1

[0796] Reaction formula:

[0797]

[0798] Material ratio:

[0799] Material Name Molecular weight Feed ratio Feeding amount mmol Heptadecan-9-ol 256.5 1eq 7.7g 30 8-Bromooctanoic acid 223 1.1 eq 7.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0800] Operation process:

[0801] Heptadecan-9-ol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 9.5 g of an oil.

[0802] Step 2: Preparation of 25-2

[0803] Reaction formula:

[0804]

[0805] Material ratio:

[0806] Material Name Molecular weight Feed ratio Feeding amount mmol 25-1 461.57 0.8eq 3.7g 8 trans-4-aminocyclohexanol 115 1eq 1.15g 10 potassium carbonate 138 1.5 eq 2.1g 15 Acetonitrile - - 50mL -

[0807] Operation process:

[0808] 25-1, trans-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask and stirred at 85°C for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.4 g of a colorless oil.

[0809] Step 3: Preparation of 25-3

[0810] Reaction formula:

[0811]

[0812] Material ratio:

[0813]

[0814]

[0815] Operation process:

[0816] 1-Nanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[0817] Step 4: Preparation of compound 25

[0818] Reaction formula:

[0819]

[0820] Material ratio:

[0821] Material Name Molecular weight Feed ratio Feeding amount mmol 25-3 349 1.2eq 1.3g 3.6 25-2 495.8 1 eq 1.5g 3 potassium carbonate 138 1.5 eq 620mg 4.5 Potassium iodide 166 0.5 eq 250mg 1.5 Acetonitrile - - 30mL -

[0822] Operation process:

[0823] 25-2, 25-3, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1 g of a colorless oil.

[0824] 1 H 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).

[0825] MS (ES+) m / z): 764.0 (M) + .

[0826] Preparation Example 19 Preparation of Compound 26

[0827] Step 1: Preparation of 26-1

[0828] Reaction formula:

[0829]

[0830] Material ratio:

[0831] Material Name Molecular weight Feed ratio Feeding amount mmol Heptadecan-9-ol 256.5 1eq 7.7g 30 8-Bromooctanoic acid 223 1.1 eq 7.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0832] Operation process:

[0833] Heptadecan-9-ol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 9.5 g of an oil.

[0834] Step 2: Preparation of 26-2

[0835] Reaction formula:

[0836]

[0837] Material ratio:

[0838] Material Name Molecular weight Feed ratio Feeding amount mmol 26-1 461.57 0.8eq 3.7g 8 cis-4-aminocyclohexanol 115 1eq 1.15g 10 potassium carbonate 138 1.5 eq 2.1g 15 Acetonitrile - - 50mL -

[0839] Operation process:

[0840] 26-1, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.4 g of a colorless oil.

[0841] Step 3: Preparation of 26-3

[0842] Reaction formula:

[0843]

[0844] Material ratio:

[0845] Material Name Molecular weight Feed ratio Feeding amount mmol 1-nonanol 144 1eq 4.3g 30 8-Bromooctanoic acid 223 1.1 eq 7.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0846] Operation process:

[0847] 1-Nanol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[0848] Step 4: Preparation of compound 26

[0849] Reaction formula:

[0850]

[0851] Material ratio:

[0852] Material Name Molecular weight Feed ratio Feeding amount mmol 26-3 349 1.2eq 1.3g 3.6 26-2 495.8 1 eq 1.5g 3 potassium carbonate 138 1.5 eq 620mg 4.5 Potassium iodide 166 0.5 eq 250mg 1.5 Acetonitrile - - 30mL -

[0853] Operation process:

[0854] 26-2, 26-3, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.2 g of a colorless oil.

[0855] 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).

[0856] MS (ES+) m / z): 764.0 (M) + .

[0857] Preparation Example 20 Preparation of Compound 27

[0858] Step 1: Preparation of 27-1

[0859] Reaction formula:

[0860]

[0861] Material ratio:

[0862] Material Name Molecular weight Feed ratio Feeding amount mmol Heptadecan-9-ol 256.5 1eq 7.7g 30 8-Bromooctanoic acid 223 1.1 eq 7.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0863] Operation process:

[0864] Heptadecan-9-ol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 9.5 g of an oil.

[0865] Step 2: Preparation of 27-2

[0866] Reaction formula:

[0867]

[0868] Material ratio:

[0869] Material Name Molecular weight Feed ratio Feeding amount mmol 27-1 461.57 0.8eq 3.7g 8 trans-4-aminocyclohexanol 115 1eq 1.15g 10 potassium carbonate 138 1.5 eq 2.1g 15 Acetonitrile - - 50mL -

[0870] Operation process:

[0871] 27-1, trans-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.4 g of a colorless oil.

[0872] Step 3: Preparation of 27-3

[0873] Reaction formula:

[0874]

[0875] Material ratio:

[0876] Material Name Molecular weight Feed ratio Feeding amount mmol 1-Undecanol 172 1eq 5.2g 30 6-Bromohexanoic acid 195 1.1 eq 6.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0877] Operation process:

[0878] 1-Undecanol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7.5 g of an oil.

[0879] Step 4: Preparation of compound 27

[0880] Reaction formula:

[0881]

[0882] Material ratio:

[0883] Material Name Molecular weight Feed ratio Feeding amount mmol 27-3 349 1.2eq 1.3g 3.6 27-2 495.8 1 eq 1.5g 3 potassium carbonate 138 1.5 eq 620mg 4.5 Potassium iodide 166 0.5 eq 250mg 1.5 Acetonitrile - - 30mL -

[0884] Operation process:

[0885] 27-2, 27-3, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.2 g of a colorless oil.

[0886] 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).

[0887] MS (ES+) m / z): 764.0 (M) + .

[0888] Preparation Example 21 Preparation of Compound 28

[0889] Step 1: Preparation of 28-1

[0890] Reaction formula:

[0891]

[0892] Material ratio:

[0893] Material Name Molecular weight Feed ratio Feeding amount mmol Heptadecan-9-ol 256.5 1eq 7.7g 30 8-Bromooctanoic acid 223 1.1 eq 7.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0894] Operation process:

[0895] Heptadecan-9-ol, 8-bromooctanoic acid, EDCI, DMAP, and DCM were added to a reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 9.5 g of an oil.

[0896] Step 2: Preparation of 28-2

[0897] Reaction formula:

[0898]

[0899] Material ratio:

[0900] Material Name Molecular weight Feed ratio Feeding amount mmol 28-1 461.57 0.8eq 3.7g 8 cis-4-aminocyclohexanol 115 1eq 1.15g 10 potassium carbonate 138 1.5 eq 2.1g 15 Acetonitrile - - 50mL -

[0901] Operation process:

[0902] 28-1, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.4 g of a colorless oil.

[0903] Step 3: Preparation of 28-3

[0904] Reaction formula:

[0905]

[0906] Material ratio:

[0907] Material Name Molecular weight Feed ratio Feeding amount mmol 1-Undecanol 172 1eq 5.2g 30 6-Bromohexanoic acid 195 1.1 eq 6.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0908] Operation process:

[0909] 1-Undecanol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7.5 g of an oil.

[0910] Step 4: Preparation of compound 28

[0911] Reaction formula:

[0912]

[0913] Material ratio:

[0914] Material Name Molecular weight Feed ratio Feeding amount mmol 28-3 349 1.2eq 1.3g 3.6 28-2 495.8 1 eq 1.5g 3 potassium carbonate 138 1.5 eq 620mg 4.5 Potassium iodide 166 0.5 eq 250mg 1.5 Acetonitrile - - 30mL -

[0915] Operation process:

[0916] 28-2, 28-3, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.3 g of a colorless oil.

[0917] 1 H 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).

[0918] MS (ES+) m / z): 764.1 (M) + .

[0919] Preparation Example 22 Preparation of Compound 30

[0920] Step 1: Synthesis of 1-({6-[(6-{[2-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)[(1r,4r)-4-hydroxycyclohexyl]amino]hexanoyl}oxy)dodec-2-yloctyl ester

[0921] Reaction formula:

[0922]

[0923] Material ratio:

[0924]

[0925] Operation process:

[0926] 1-[(6-bromohexanoyl)oxy]dodec-2-yl octyl ester (2.00 g, 3.96 mmol) was dissolved in acetonitrile (15 mL), and (1r,4r)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 20 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({6-[(6-{[2-(octanoyloxy)dodecyl]oxy}-6-oxyylidenehexyl)[(1r,4r)-4-hydroxycyclohexyl]amino]hexanoyl}oxy)dodec-2-yl octyl ester (1.00 g, 25.9% yield) as a yellow liquid.

[0927] 1 H 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.

[0928] LCMS:RT=2.172,m / z 965.4[M+H] + .

[0929] Preparation Example 23 Preparation of Compound 8

[0930] Step 1: Preparation of 8-1

[0931] Reaction formula:

[0932]

[0933] Material ratio:

[0934] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 19.5g 100 1,2-Epoxydodecane 184 1eq 18.4g 100 Ferric chloride 162 0.05 eq 800mg 5 Pyridine 79 0.025 eq 200mg 2.5

[0935] Operation process:

[0936] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[0937] Step 2: Preparation of 8-2

[0938] Reaction formula:

[0939]

[0940] Material ratio:

[0941] Material Name Molecular weight Feed ratio Feeding amount mmol 8-1 379.38 1eq 3.45g 9.1 Octanoic acid 144.21 1 eq 1.3g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[0942] Operation process:

[0943] 8-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated aqueous sodium bicarbonate solution, and once with 100 mL of saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 4 g of an oil.

[0944] Step 3: Preparation of compound 8

[0945] Reaction formula:

[0946]

[0947] Material ratio:

[0948] Material Name Molecular weight Feed ratio Feeding amount mmol 8-2 505 2.5eq 2.5g 5 3-Aminocyclobutanol 87 1 eq 174mg 2 potassium carbonate 138 3 eq 830mg 6 Potassium iodide 166 1 eq 330mg 2 Acetonitrile - - 30mL -

[0949] Operation process:

[0950] 8-2,3-Aminocyclobutanol, K2CO3, KI, and acetonitrile were added to a reaction flask and heated to 75°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1 g of a colorless oil.

[0951] 1H 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).

[0952] MS (ES+) m / z): 935.9 (M) + .

[0953] Preparation Example 24 Preparation of Compound 15

[0954] Step 1: Preparation of 15-1

[0955] Reaction formula:

[0956]

[0957] Material ratio:

[0958] Material Name Molecular weight Feed ratio Feeding amount mmol p-Nitrophenyl chloroformate 201.56 1.2eq 3g 15 7-Bromoheptanol 195 1 eq 2.4g 12.5 Pyridine 79 1.5 eq 1.5g 18.8 DCM - - 50mL -

[0959] Operation process:

[0960] To a reaction flask, p-nitrophenyl chloroformate, 7-bromoheptanol, and DCM were added dropwise under nitrogen at room temperature. Pyridine was added dropwise over approximately 5 minutes. After the addition was complete, the reaction was stirred at room temperature for 16 hours. TLC (PE:EA = 6:1) revealed an Rf value of 0.5 for the product. The reaction solution was concentrated and purified by column chromatography to yield 4 g of an oil.

[0961] Step 2: Preparation of 15-2

[0962] Reaction formula:

[0963]

[0964] Material ratio:

[0965] Material Name Molecular weight Feed ratio Feeding amount mmol 15-1 360 1eq 4g 11.1 Heptadecan-9-ol 256 4eq 11.4g 44.4 Pyridine 79 1.2 eq 1.1g 13.3 DMAP 122 0.2eq 300mg 2.2 DCM - - 100mL -

[0966] Operation process:

[0967] 15-1, heptadecan-9-ol, pyridine, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 16 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was concentrated and purified by column chromatography to yield 3.5 g of an oil.

[0968] Step 3: Preparation of 15-3

[0969] Reaction formula:

[0970]

[0971] Material ratio:

[0972] Material Name Molecular weight Feed ratio Feeding amount mmol 1-Undecanol 172 1eq 5.2g 30 6-Bromohexanoic acid 195 1.1 eq 6.4g 33 EDCI 192 1.5eq 8.6g 45 DMAP 122 0.15 eq 550mg 4.5 DCM - - 150mL -

[0973] Operation process:

[0974] 1-Undecanol, 6-bromohexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated sodium bicarbonate solution and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7.5 g of an oil.

[0975] Step 4: Preparation of 15-4

[0976] Reaction formula:

[0977]

[0978] Material ratio:

[0979] Material Name Molecular weight Feed ratio Feeding amount mmol 15-3 349 0.8eq 5.6g 16 cis-4-aminocyclohexanol 115 1 eq 2.3g 20 potassium carbonate 138 1.5 eq 4.2g 30 Acetonitrile - - 80mL -

[0980] Operation process:

[0981] 15-3, cis-4-aminocyclohexanol, K2CO3, and acetonitrile were added to a reaction flask and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.4 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 3.5 g of a colorless oil.

[0982] Step 5: Preparation of compound 15

[0983] Reaction formula:

[0984]

[0985] Material ratio:

[0986]

[0987]

[0988] Operation process:

[0989] To a reaction flask, 15-2, 15-4, K2CO3, KI, and acetonitrile were added and heated to 85°C with stirring for 16 hours. TLC (DCM:MeOH = 10:1) revealed an Rf value of 0.5 for the product. The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.2 g of a colorless oil.

[0990] 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).

[0991] MS (ES+) m / z): 780.0 (M) + .

[0992] Preparation Example 25 Preparation of Compound 23

[0993] Step 1: Synthesis of dioctyl propylene glycol ester

[0994] Reaction formula:

[0995]

[0996] Material ratio:

[0997]

[0998] Operation process:

[0999] Dissolve octan-1-ol (5.01 g, 38.4 mmol) in dichloromethane (50.0 mL). Add 1,3-propanedioic 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) sequentially. Incubate at 25°C under nitrogen for 12 hours. Monitor the reaction by TLC. Concentrate the reaction mixture. Column chromatography yields the crude product (5.00 g, 79.2% yield) of dioctyl propanediol.

[1000] Step 2: Synthesis of dioctyl-2-(4-benzyloxybutyl)propanediol

[1001] Reaction formula:

[1002]

[1003] Material ratio:

[1004]

[1005]

[1006] Operation process:

[1007] Dissolve dioctyl propanediol (5.00 g, 15.2 mmol) and 4-bromobutoxymethylbenzene (3.70 g, 15.2 mmol) in acetonitrile (10.0 mL). Add tetrabutylammonium bromide (491 mg, 1.52 mmol) and potassium carbonate (2.31 g, 16.7 mmol) sequentially. Incubate at 60°C under nitrogen for 4 hours. TLC analysis indicates the reaction is complete, indicating the formation of a large amount of product. Filter and concentrate the product. Column chromatography yields the crude product as a colorless liquid (3.200 g, 42.8% yield).

[1008] Step 3: Synthesis of dioctyl-2-(4-hydroxybutyl)propanediol

[1009] Reaction formula:

[1010]

[1011] Material ratio:

[1012] Material Name Molecular weight Feed ratio Feeding amount mmol Dioctyl-2-(4-benzyloxybutyl)propanediol 490 1.00 eq 3.10g 6.32 Palladium / Carbon 106 0.10eq 0.672g 0.631 hydrogen - (40Psi) - - Methanol - - 30.0mL -

[1013] Operation process:

[1014] Dissolve dioctyl-2-(4-benzyloxybutyl)propanediol (3.10 g, 6.32 mmol) in methanol (30.0 mL). Add palladium / carbon (0.672 g, 0.63 mmol) under argon. Incubate at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Monitor the reaction by TLC. Filter the reaction mixture and concentrate. The crude product is dried and purified by column chromatography to yield dioctyl-2-(4-hydroxybutyl)propanediol (2.51 g, 99% yield) as a colorless liquid.

[1015] Step 4: Synthesis of dioctyl-2-[4-(p-toluenesulfonyloxy)butyl]propanediol

[1016] Reaction formula:

[1017]

[1018] Material ratio:

[1019] Material Name Molecular weight Feed ratio Feeding amount mmol Dioctyl-2-(4-hydroxybutyl)propanediol 401 1.00 eq 1.00g 2.50 4-Methylbenzenesulfonyl chloride 190 1.50 eq 713mg 3.74 Triethylamine 101 3.00 eq 757mg 7.49 4-Dimethylaminopyridine 122 0.10eq 30.5mg 0.25 dichloromethane - - 10.0mL -

[1020] Operation process:

[1021] Dissolve dioctyl 2-(4-hydroxybutyl)propanediol (1.00 g, 2.50 mmol) in dichloromethane (10.0 mL). Add triethylamine (757 mg, 7.49 mmol) and 4-dimethylaminopyridine (30.5 mg, 0.25 mmol) sequentially. Finally, add 4-methylbenzenesulfonyl chloride (713 mg, 3.74 mmol) dropwise at 0°C under nitrogen. The reaction is allowed to proceed at 25°C for 12 hours. Completion is monitored by TLC, and the reaction solution is concentrated. The crude product is purified by column chromatography to afford dioctyl 2-[4-(p-toluenesulfonyloxy)butyl]propanediol (1.30 g, 93% yield).

[1022] Step 5: Synthesis of [3-(6-bromohexanoyloxy)-2-octaoxo-propyl]octyl ester

[1023] Reaction formula:

[1024]

[1025] Material ratio:

[1026]

[1027] Operation process:

[1028] Dissolve dioctyl-2-[4-(p-toluenesulfonyloxy)butyl]propanediol (1.00 g, 1.80 mmol) in acetonitrile (10.0 mL). 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 mixture was reacted at 85°C under nitrogen for 20 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a yellow liquid, [dioctyl-2-[4-[(4-hydroxycyclohexyl)-[8-(1-octylnonaoxo)-8-oxyylidene-octyl]amino]butyl]propanediol (70.0 mg, 9.00% yield).

[1029] 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.

[1030] LCMS: RT=1.949, m / z=879.2.[M+H] + .

[1031] Preparation Example 26 Preparation of Compound 24

[1032] Step 1: Preparation of 24-1

[1033] Reaction formula:

[1034]

[1035] Material ratio:

[1036]

[1037]

[1038] Operation process:

[1039] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 18 g of an oil.

[1040] Step 2: Preparation of 24-2

[1041] Reaction formula:

[1042]

[1043] Material ratio:

[1044] Material Name Molecular weight Feed ratio Feeding amount mmol 24-1 379.38 1 eq 3.45g 9.1 n-Hexanoic acid 116 1 eq 1.1g 9.1 EDCI 192 1.5eq 2.7g 13.7 DMAP 122 0.15 eq 200mg 1.4 DCM - - 60mL -

[1045] Operation process:

[1046] 24-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at room temperature for 2 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was diluted with 100 mL of dichloromethane, washed once with 100 mL of saturated sodium bicarbonate solution, and once with 100 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 3.4 g of an oil.

[1047] Step 3: Preparation of compound 24

[1048] Reaction formula:

[1049]

[1050] Material ratio:

[1051] Material Name Molecular weight Feed ratio Feeding amount mmol 24-2 505.58 2.2eq 3.4g 6.7 cis-4-aminocyclohexanol 115.17 1 eq 345mg 3 potassium carbonate 138 3 eq 1.3g 9 Potassium iodide 166 1 eq 500mg 3 Acetonitrile - - 40mL -

[1052] Operation process:

[1053] 24-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 20:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 1.8 g of an oil.

[1054] 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).

[1055] MS (ES+) m / z): 907.9 (M) + .

[1056] Preparation Example 27 Preparation of Compound 29

[1057] Step 1: Synthesis of (3-benzyloxy-2-octaoxo-propyloxy)methylbenzene

[1058] Reaction formula:

[1059]

[1060] Material ratio:

[1061] Material Name Molecular weight Feed ratio Feeding amount mmol 1,3-Dibenzyloxypropan-2-ol 272 1.00 eq 3.00g 11.0 1-iodooctane 240 1.20eq 3.17g 13.2 Sodium hydride (60%) 24 1.50 eq 661mg 16.5 N,N-dimethylformamide - - 30.0mL -

[1062] Operation process:

[1063] 1,3-Dibenzyloxypropan-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 under nitrogen for 0.5 hours. 1-Iodooctane (3.17 g, 13.2 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was allowed to proceed at 20°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was extracted and concentrated. Column chromatography of the crude product afforded (3-benzyloxy-2-octaoxo-propoxy)methylbenzene (1.70 g, 40.0% yield) as a colorless liquid.

[1064] Step 2: Synthesis of 2-octaoxopropane-1,3-diol

[1065] Reaction formula:

[1066]

[1067] Material ratio:

[1068] Material Name Molecular weight Feed ratio Feeding amount mmol 3-Benzyloxy-2-octaoxo-propyloxy)methylbenzene 385 1.00 eq 1.70g 4.42 Palladium / Carbon 106 0.20eq 0.941g 0.88 hydrogen - (40Psi) - - Methanol - - 20.0mL -

[1069] Operation process:

[1070] Dissolve 3-benzyloxy-2-octaoxo-propoxymethylbenzene (1.70 g, 4.42 mmol) in methanol (20.0 mL). Add palladium / carbon (0.941 g, 0.88 mmol) under argon. Incubate at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Monitor the reaction by TLC. Filter the reaction mixture and concentrate. The crude product is dried and purified by column chromatography to yield 2-octaoxopropane-1,3-diol (900 mg, 99% yield), a colorless liquid.

[1071] Step 3: Synthesis of (3-hydroxy-2-octaoxo-propyl) octyl ester

[1072] Reaction formula:

[1073]

[1074] Material ratio:

[1075]

[1076] Operation process:

[1077] Dissolve 2-octaoxopropane-1,3-diol (900 mg, 4.41 mmol) in dichloromethane (10.0 mL). Add n-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) sequentially. Incubate at 25°C under nitrogen for 12 hours. Completion of the reaction is monitored by TLC. The reaction solution is concentrated. Column chromatography yields the crude product as a colorless liquid (3-hydroxy-2-octaoxopropyl)octyl ester (700 mg, 48.0% yield).

[1078] Step 4: Synthesis of [3-(6-bromohexanoyloxy)-2-octaoxo-propyl]octyl ester

[1079] Reaction formula:

[1080]

[1081] Material ratio:

[1082]

[1083] Operation process:

[1084] Dissolve (3-hydroxy-2-octaoxo-propyl)octyl ester (700 mg, 2.12 mmol) in dichloromethane (10.0 mL). 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 mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was concentrated. Column chromatography of the crude product afforded [3-(6-bromohexanoyloxy)-2-octaoxo-propyl]octyl ester (520 mg, 48.3% yield) as a colorless liquid.

[1085] Step 5: Synthesis of [3-[6-[(4-hydroxycyclohexyl)-[6-(3-octanoyloxy-2-octaoxo-propyloxy)-6-oxyylidene-hexyl]amino]hexanoyloxy]-2-octaoxo-propyl]octyl ester

[1086] Reaction formula:

[1087]

[1088] Material ratio:

[1089]

[1090] Operation process:

[1091] Dissolve [3-(6-bromohexanoyloxy)-2-octaoxo-propyl]octyl ester (1.20 g, 2.36 mmol) in acetonitrile (15.00 mL), and add 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) in sequence. The mixture is reacted at 80°C under nitrogen for 12 hours. The reaction is monitored by TLC, filtered, and concentrated. Purification by column chromatography affords [3-[6-[(4-hydroxycyclohexyl)-[6-(3-octanoyloxy-2-octaoxo-propoxy)-6-oxyylidene-hexyl]amino]hexanoyloxy]-2-octaoxo-propyl]octyl ester (700 mg, 30.0% yield), a colorless liquid.

[1092] 1 H 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.

[1093] LCMS: RT=2.429, m / z=996.8.[M+H] + .

[1094] Preparation Example 28 Preparation of Compound 11

[1095] Step 1: Synthesis of 1,3-bis(benzyloxy)propan-2-yl 8-bromooctyl ester

[1096] Reaction formula:

[1097]

[1098] Material ratio:

[1099]

[1100] Operation process:

[1101] Dissolve 1,3-bis(benzyloxy)propan-2-ol (24.00 g, 7.34 mmol) and 8-bromooctanoic acid (1.97 g, 8.81 mmol) in dichloromethane (20 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.11 g, 11.02 mmol) and 4-dimethylaminopyridine (179 mg, 1.47 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring revealed the formation of new spots. The reaction solution was concentrated. Column chromatography of the crude product afforded 1,3-bis(benzyloxy)propan-2-yl 8-bromooctyl ester (3.40 g, 96.9% yield) as a colorless liquid.

[1102] Step 2: Synthesis of 1,3-dihydroxypropane-2-yl 8-bromooctyl ester

[1103] Reaction formula:

[1104]

[1105] Material ratio:

[1106] Material Name Molecular weight Feed ratio Feeding amount mmol 1,3-Bis(benzyloxy)propan-2-yl 8-bromooctyl ester 477 1.0 eq 3.40g 7.12 Palladium on carbon 105 0.2eq 1.52g 1.42 Methanol - - 34mL -

[1107] Operation process:

[1108] Dissolve 1,3-di(benzyloxy)propan-2-yl 8-bromooctyl ester (3.40 g, 7.12 mmol) in methanol (34 mL), add palladium on carbon (1.52 g, 1.42 mmol), and react under hydrogen at 35°C, 40 psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1,3-dihydroxypropan-2-yl 8-bromooctyl ester (1.34 g, 63.3% yield), a yellow liquid.

[1109] Step 3: Synthesis of 1,3-bis(hexanoyloxy)propan-2-yl 8-bromooctyl ester

[1110] Reaction formula:

[1111]

[1112] Material ratio:

[1113]

[1114] Operation process:

[1115] Dissolve 1,3-dihydroxypropan-2-yl 8-bromooctyl ester (1.34 g, 4.51 mmol) and hexanoic acid (1.05 g, 9.02 mmol) in dichloromethane (15 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.59 g, 13.5 mmol) and 4-dimethylaminopyridine (220 mg, 1.80 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. Column chromatography of the crude product afforded 1,3-di(hexanoyloxy)propan-2-yl 8-bromooctyl ester (2.00 g, 68.7% yield) as a colorless liquid.

[1116] Step 4: Synthesis of undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester

[1117] Reaction formula:

[1118]

[1119] Material ratio:

[1120] Material Name Molecular weight Feed ratio Feeding amount mmol Undecyl 6-bromohexyl ester 348 1.0 eq 2.00g 5.72 (1s,4s)-4-Aminocyclohexan-1-ol 115 2.0eq 1.32g 11.4 Triethylamine 101 2.0eq 1.16g 11.4 Acetonitrile - - 20mL -

[1121] Operation process:

[1122] Undecyl 6-bromohexyl ester (2.00 g, 5.72 mmol) was dissolved in acetonitrile (20 mL). (1s,4s)-4-aminocyclohexan-1-ol (1.32 g, 11.4 mmol) and triethylamine (1.32 g, 11.4 mmol) were added to the reaction mixture. The mixture was reacted at 40°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to afford undecyl 6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexyl ester (910 mg, 41.1% yield) as a pink liquid.

[1123] Step 5: Synthesis of 1,3-bis(hexanoyloxy)propan-2-yl 8-{[6-oxyylidene-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester

[1124] Reaction formula:

[1125]

[1126] Material ratio:

[1127]

[1128] Operation process:

[1129] 1,3-Di(hexanoyloxy)propan-2-yl 8-bromooctyl ester (1.17 g, 2.36 mmol) was dissolved in acetonitrile (10 mL). 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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1,3-Di(hexanoyloxy)propan-2-yl 8-{[6-oxyidene-6-(undecyloxy)hexyl][(1s,4s)-4-hydroxycyclohexyl]amino}octyl ester (300 mg, 70.4% yield) as a yellow liquid.

[1130] 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.

[1131] LCMS:RT=0.740,m / z 797.3[M+H] + .

[1132] Preparation Example 29 Preparation of Compound 16

[1133] Step 1: Synthesis of 4-nitrophenylpentadecan-8-yl carbonate

[1134] Reaction formula:

[1135]

[1136] Material ratio:

[1137] Material Name Molecular weight Feed ratio Feeding amount mmol Pentadecane-8-ol 228 1.00 eq 1.00g 4.38 4-Nitrophenyl chloride 200 1.20eq 1.06g 5.25 4-Dimethylaminopyridine 122 2.00eq 1.07g 8.76 dichloromethane - - 10.0mL - dichloromethane - - 10.0mL -

[1138] Operation process:

[1139] 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. Chlorophenyl ester (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 mixture was reacted at 25°C for 12 hours. The reaction was monitored by TLC and the mixture was used directly in the next step to obtain a yellow liquid 4-nitrophenylpentadecan-8-yl carbonate (1.72 g, 100% yield).

[1140] Step 2: Synthesis of 6-bromohexyl 3-(benzyloxy)propyl ester

[1141] Reaction formula:

[1142]

[1143] Material ratio:

[1144]

[1145] Operation process:

[1146] 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). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.19 g, 16.6 mmol) and 4-dimethylaminopyridine (135 mg, 1.11 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. Purification by column chromatography gave 6-bromohexyl 3-(benzyloxy)propyl ester (2.67 g, 70.0% yield) as a colorless liquid.

[1147] Step 3: Synthesis of 6-bromohexyl 3-hydroxypropyl ester

[1148] Reaction formula:

[1149]

[1150] Material ratio:

[1151] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexyl 3-(benzyloxy)propyl ester 402 1.00 eq 2.10g 6.12 Boron trichloride 117 2.00eq 1.43g 12.2 dichloromethane - - 21.0mL -

[1152] Operation process:

[1153] 6-Bromohexyl 3-(benzyloxy)propyl ester (2.10 g, 6.12 mmol) was dissolved in dichloromethane (21.0 mL). Boron trichloride (1 M, 12.2 mL) was added at 0°C. The mixture was reacted at 0°C for 1 hour under nitrogen. TLC revealed the formation of new spots. After adjusting the pH to neutral, the mixture was extracted with ethyl acetate and the organic phase was concentrated. Purification by column chromatography afforded 6-bromohexyl 3-hydroxypropyl ester (1.50 g, 96.8% yield) as a colorless liquid.

[1154] Step 4: Synthesis of 6-bromohexyl 3-{[(pentadecan-8-oxy)carbonyl]oxy}propyl ester

[1155] Reaction formula:

[1156]

[1157] Material ratio:

[1158] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexyl 3-hydroxypropyl ester 252 1.00 eq 1.00g 3.95 4-Nitrophenylpentadecan-8-yl carbonate 393 1.00eq 1.55g 3.95 dichloromethane - - 10.0mL -

[1159] Operation process:

[1160] 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 dropwise at 25°C. The mixture was reacted for 12 hours at 25°C. The reaction was monitored by TLC. The mixture was purified by column chromatography to give a colorless liquid 6-bromohexyl 3-{[(pentadecan-8-oxy)carbonyl]oxy}propyl ester (1.30 g, 64.8% yield).

[1161] Step 5: Synthesis of 6-({6-[(3-{[(pentadecan-8-oxy)carbonyl]oxy}propionyl)oxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl 3-{[(pentadecan-8-oxy)carbonyl]oxy}propyl ester

[1162] Reaction formula:

[1163]

[1164] Material ratio:

[1165]

[1166] Operation process:

[1167] Dissolve 6-bromohexyl 3-{[(pentadecan-8-oxy)carbonyl]oxy}propyl ester (940 mg, 1.85 mmol) in acetonitrile (10.0 mL), and add (1s,4s)-4-aminocyclohexan-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) in sequence. Finally, react at 75°C under nitrogen protection for 12 hours. The reaction is completed after monitoring by TLC. The mixture is filtered and concentrated. Purification by column chromatography gave colorless liquid 6-({6-[(3-{[(pentadecan-8-oxy)carbonyl]oxy}propionyl)oxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl 3-{[(pentadecan-8-oxy)carbonyl]oxy}propyl ester (163 mg, 9.09% yield).

[1168] 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.

[1169] LCMS:RT=2.490,m / z 969.6[M+H] + .

[1170] Preparation Example 30 Preparation of Compound 18

[1171] Step 1: Synthesis of 1-(benzyloxy)dodecan-2-ol

[1172] Reaction formula:

[1173]

[1174] Material ratio:

[1175] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecan-2-ol 292 1.00 eq 4.00g 10.9 1-iodooctane 240 1.20eq 3.94g 16.4 Sodium hydride (60%) 24 1.50 eq 656mg 16.4 N,N-dimethylformamide - - 50.0mL -

[1176] Operation process:

[1177] 1-(Benzyloxy)dodecan-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 under nitrogen for 0.5 hours. 1-Iodooctane (3.94 g, 13.2 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was allowed to proceed at 20°C under nitrogen for 12 hours. The reaction was monitored by TLC, and the reaction solution was extracted and concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodecan-2-ol (1.50 g, 34.0% yield) as a colorless liquid.

[1178] Step 2: Synthesis of 2-(octyloxy)dodecan-1-ol

[1179] Reaction formula:

[1180]

[1181] Material ratio:

[1182] Material Name Molecular weight Feed ratio Feeding amount mmol ({[2-(Octyloxy)dodecyl]oxy}methyl)benzene 404 1.00 eq 1.50g 3.71 Palladium / Carbon 106 0.10eq 0.394g 0.371 hydrogen - (40Psi) - - Methanol - 10.0mL -

[1183] Operation process:

[1184] Dissolve ({[2-(Octyloxy)dodecyl]oxy}methyl)benzene (1.50 g, 3.71 mmol) in methanol (30.0 mL). Add palladium / carbon (0.394 g, 0.37 mmol) under argon. Incubate at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Monitor the reaction by TLC. Filter the reaction mixture and concentrate. The crude product is dried and purified by column chromatography to yield 2-(octyloxy)dodecan-1-ol (1.17 g, 99.9% yield) as a colorless liquid.

[1185] Step 3: Synthesis of 2-(octyloxy)dodecyl 6-bromohexyl ester

[1186] Reaction formula:

[1187]

[1188] Material ratio:

[1189]

[1190] Operation process:

[1191] Dissolve 2-(Octyloxy)dodecan-1-ol (1.17 g, 3.72 mmol) in dichloromethane (10.0 mL). 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) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. Column chromatography of the crude product afforded 2-(Octyloxy)dodecyl 6-bromohexyl ester (1.70 g, 93.0% yield) as a colorless liquid.

[1192] Step 4: Synthesis of 2-(octyloxy)dodecyl 6-[(6-{[2-(octyloxy)dodecyl]oxy}-6-oxyylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl ester

[1193] Reaction formula:

[1194]

[1195] Material ratio:

[1196] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Octyloxy)dodecyl 6-bromohexyl ester 490 1.0 eq 1.50g 3.05 (1s,4s)-4-Aminocyclohexan-1-ol 115 0.45 eq 158mg 1.37 potassium carbonate 138 3.0eq 1.27g 9.15 Potassium iodide 166 1.5 eq 759mg 4.58 Acetonitrile - - 10mL - Tetrahydrofuran - - 5mL -

[1197] Operation process:

[1198] 2-(Octyloxy)dodecyl 6-bromohexyl ester (1.50 g, 3.05 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 20 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(Octyloxy)dodecyl 6-[(6-{[2-(Octyloxy)dodecyl]oxy}-6-oxyylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl ester (700 mg, 24.15%) as a yellow liquid.

[1199] 1H 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).

[1200] LCMS:RT=2.469,m / z 937.9[M+H] + .

[1201] Preparation Example 31 Preparation of Compound 31

[1202] Step 1: Synthesis of 6-[(6-{[2-(octanoyloxy)dodecanoyl]oxy}hexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl 2-(octanoyloxy)dodecyl ester

[1203] Reaction formula:

[1204]

[1205] Material ratio:

[1206]

[1207]

[1208] Operation process:

[1209] 6-Bromohexyl 2-(octanoyloxy)dodecyl ester (1.50 g, 2.97 mmol) was dissolved in acetonitrile (10 mL). (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 6-[(6-{[2-(octanoyloxy)dodecanoyl]oxy}hexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexyl 2-(octanoyloxy)dodecyl ester (580 mg, 20.2% yield) as a yellow liquid.

[1210] 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.

[1211] LCMS:RT=2.217,m / z 964.8[M+H] + .

[1212] Preparation Example 32 Preparation of Compound 32

[1213] Step 1: Synthesis of ({[1-(benzyloxy)-3-[(6-bromohexyl)oxy]propan-2-yl]oxy}methyl)benzene

[1214] Reaction formula:

[1215]

[1216] Material ratio:

[1217]

[1218] Operation process:

[1219] 2,3-Bis(benzyloxy)propan-1-ol (900 mg, 3.30 mmol) was dissolved in N,N-dimethylformamide (30 mL). Sodium hydroxide (60%) (198 mg, 4.96 mmol) was added at 0°C and allowed to react for 0.5 hours. 6-Bromohexan-1-ol (1.21 g, 4.96 mmol) was then added and allowed to react at 25°C under nitrogen for 12 hours. TLC revealed the formation of new spots. The reaction solution was quenched with 300 mL of water and washed twice with 300 mL of ethyl acetate. The organic phase was concentrated. Column chromatography of the crude product afforded ({[1-(benzyloxy)-3-[(6-bromohexyl)oxy]propan-2-yl]oxy}methyl)benzene (740 mg, 51.4% yield) as a colorless liquid.

[1220] Step 2: Synthesis of 3-[(6-bromohexyl)oxy]propane-1,2-diol

[1221] Reaction formula:

[1222]

[1223] Material ratio:

[1224]

[1225] Operation process:

[1226] Dissolve ({[1-(benzyloxy)-3-[(6-bromohexyl)oxy]propan-2-yl]oxy}methyl)benzene (740 mg, 1.70 mmol) in dichloromethane (8 mL). Add boron trichloride (597 mg, 5.10 mmol) in a 1M CHCl solution at 0°C. The reaction was allowed to react at 0°C for 1 hour. Completion was monitored by TLC. The reaction was quenched with 10 mL of aqueous sodium bicarbonate solution and extracted with dichloromethane twice, each time with 10 mL. The organic phase was dried, filtered, and concentrated. Purification by column chromatography afforded 3-[(6-bromohexyl)oxy]propane-1,2-diol (240 mg, 55.3% yield), a colorless liquid.

[1227] Step 3: Synthesis of 1-[(6-bromohexyl)oxy]-3-(octanoyloxy)propan-2-yloctyl ester

[1228] Reaction formula:

[1229]

[1230] Material ratio:

[1231]

[1232] Operation process:

[1233] 3-[(6-bromohexyl)oxy]propane-1,2-diol (240 mg, 0.940 mmol) and octanoic acid (298 mg, 2.07 mmol) were dissolved in dichloromethane (3 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (540 mg, 2.82 mmol) and 4-dimethylaminopyridine (45.9 mg, 0.376 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-[(6-bromohexyl)oxy]-3-(octanoyloxy)propan-2-yloctyl ester (380 mg, 79.60% yield) as a colorless liquid.

[1234] Step 4: Synthesis of 1-{[6-({6-[2,3-di(octanoyloxy)propoxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl]oxy}-3-(octanoyloxy)propan-2-yloctyl ester

[1235] Reaction formula:

[1236]

[1237] Material ratio:

[1238]

[1239] Operation process:

[1240] 1-[(6-bromohexyl)oxy]-3-(octanoyloxy)propan-2-yl octyl ester (300 mg, 0.591 mmol) was dissolved in acetonitrile (2 mL), and (1s,4s)-4-aminocyclohexan-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) were added sequentially. The mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-{[6-({6-[2,3-di(octanoyloxy)propoxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl]oxy}-3-(octanoyloxy)propan-2-yl octyl ester (130 mg, 22.4% yield) as a yellow liquid.

[1241] 1 H 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.

[1242] LCMS:RT=1.643,m / z 968.8[M+H] + .

[1243] Preparation Example 34 Preparation of Compound 53

[1244] Step 1: Synthesis of 2-hydroxydodecyl 6-bromohexyl ester

[1245] Reaction formula:

[1246]

[1247] Material ratio:

[1248] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Decyloxapropane 184 1.0 eq 4.00g 21.7 6-Bromohexanoic acid 194 1.1eq 4.66g 23.8 Ferric chloride 160 0.025eq 88.0mg 0.542 Pyridine 79 0.028eq 48.0mg 0.607

[1249] Operation process:

[1250] To 2-decyloxirane (4.00 g, 21.7 mmol) was added 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). The mixture was reacted at 40°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The organic phase was concentrated. The crude product was purified by column chromatography to afford 2-hydroxydodecyl 6-bromohexyl ester (6.40 g, 77.7% yield) as a colorless liquid.

[1251] Step 2: Synthesis of 2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl 6-bromohexyl ester

[1252] Reaction formula:

[1253]

[1254] Material ratio:

[1255]

[1256] Operation process:

[1257] 2-Hydroxydodecyl 6-bromohexyl ester (3.00 g, 7.91 mmol) and 5-(1,2-dithiolan-3-yl)pentanoic acid (1.63 g, 7.91 mmol) were dissolved in dichloromethane (30 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27 g, 11.8 mmol) and 4-dimethylaminopyridine (193 mg, 1.58 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl 6-bromohexyl ester (2.7 g, 60.15% yield) as a yellow liquid.

[1258] Step 3: Synthesis of 2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl 6-({6-[(2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl)oxy]-6-oxyylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[1259] Reaction formula:

[1260]

[1261] Material ratio:

[1262]

[1263] Operation process:

[1264] 2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl 6-bromohexyl ester (1.20 g, 2.11 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexan-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) were added in sequence. Finally, the mixture was reacted at 80 ° C under nitrogen protection for 12 hours. The reaction was completed after monitoring by TLC. The mixture was filtered and concentrated. Purification by column chromatography gave a yellow liquid 2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl 6-({6-[(2-{[5-(1,2-dithiolan-3-yl)pentanoyl]oxy}dodecyl)oxy]-6-oxoylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (430 mg, 17.36% yield).

[1265] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.16-5.03(m,2H),4.33-4.23(m,2H),4.13(brs,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),2.35(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.

[1266] LCMS:RT=2.441,m / z 1089.1[M+H] + .

[1267] Preparation Example 35 Preparation of Compound 54

[1268] Step 1: Synthesis of 3-(butylmercapto)propionic acid

[1269] Reaction formula:

[1270]

[1271] Material ratio:

[1272] Material Name Molecular weight Feed ratio Feeding amount mmol 3-Mercaptopropionic acid 106 1.0 eq 4.00g 37.6 1-Iodobutane 184 1.0eq 6.93g 37.6 potassium hydroxide 56 1.2eq 6.34g 45.2 Methanol - - 40mL -

[1273] Operation process:

[1274] 3-Mercaptopropionic acid (4.00 g, 37.6 mmol) and 1-iodobutane (6.93 g, 37.6 mmol) were dissolved in methanol (40 mL). Potassium hydroxide (6.34 g, 45.2 mmol) was added and the mixture was allowed to react at 20°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. 10 mL of water was added and the mixture was extracted with ethyl acetate. The aqueous phase was adjusted to pH 5 with 1 M hydrochloric acid and extracted twice with 20 mL of ethyl acetate each time. The organic phase was concentrated. The crude product was purified by column chromatography to yield 3-(butylmercapto)propionic acid (2.13 g, 34.8% yield), a colorless liquid.

[1275] Step 2: Synthesis of 2-{[3-(butylmercapto)propionyl]oxy}dodecyl 6-bromohexyl ester

[1276] Reaction formula:

[1277]

[1278] Material ratio:

[1279]

[1280]

[1281] Operation process:

[1282] 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). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.27 g, 11.8 mmol) and 4-dimethylaminopyridine (193 mg, 1.58 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. The reaction solution was concentrated. The crude product was purified by column chromatography to obtain 2-{[3-(butylmercapto)propionyl]oxy}dodecyl 6-bromohexyl ester (3.14 g, 75.8% yield) as a colorless liquid.

[1283] Step 3: Synthesis of 2-{[3-(butylmercapto)propionyl]oxy}dodecyl 6-({6-[(2-{[3-(butylmercapto)propionyl]oxy}dodecyl)oxy]-6-oxyylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[1284] Reaction formula:

[1285]

[1286] Material ratio:

[1287]

[1288] Operation process:

[1289] Dissolve 2-{[3-(Butylmercapto)propionyl]oxy}dodecyl 6-bromohexyl ester (1.50 g, 2.86 mmol) in acetonitrile (10 mL), and add (1s,4s)-4-aminocyclohexan-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) in sequence. Finally, react at 80 ° C under nitrogen protection for 12 hours. The reaction is completed after monitoring by TLC. The mixture is filtered and concentrated. Purification by column chromatography gave a yellow liquid 2-{[3-(butylmercapto)propionyl]oxy}dodecyl 6-({6-[(2-{[3-(butylmercapto)propionyl]oxy}dodecyl)oxy]-6-oxoylidenehexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (600 mg, 20.1% yield).

[1290] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.14-5.04(m,2H),4.30-4.22(m,2H),4.11(brs,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,4 H),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.

[1291] LCMS:RT=2.477,m / z 1001.2[M+H] + .

[1292] Preparation Example 36 Preparation of Compound 33

[1293] Step 1: Preparation of 33-1

[1294] Reaction formula:

[1295]

[1296] Material ratio:

[1297] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195 1 eq 9.75g 50 1,2-Epoxytetradecane 212 1eq 10.6g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1298] Operation process:

[1299] 6-Bromohexanoic acid, 1,2-epoxytetradecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1300] Step 2: Preparation of 33-2

[1301] Reaction formula:

[1302]

[1303] Material ratio:

[1304] Material Name Molecular weight Feed ratio Feeding amount mmol 33-1 407.43 1eq 8g 19.6 n-Decanic acid 172.27 1.1 eq 3.7g 21.6 EDCI 192 2 eq 7.5g 39.2 DMAP 122 0.2 eq 480mg 3.9 DCM - 20V 160mL -

[1305] Operation process:

[1306] To the reaction flask, 33-1, n-decanoic acid, EDCI, DMAP, and DCM were added and stirred at -5°C-0°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 200 mL of saturated sodium bicarbonate solution and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 10 g of an oil.

[1307] Step 3: Preparation of compound 33

[1308] Reaction formula:

[1309]

[1310] Material ratio:

[1311] Material Name Molecular weight Feed ratio Feeding amount mmol 33-2 561.7 1eq 4g 7.1 cis-4-aminocyclohexanol 115.17 0.4 eq 322mg 2.8 potassium carbonate 138 2eq 2g 14.2 Potassium iodide 166 1 eq 1.2g 7.1 Acetonitrile - 20V 80mL -

[1312] Operation process:

[1313] 33-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.2 g of an oil.

[1314] 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).

[1315] MS (ES+) m / z): 1076.8 (M) + .

[1316] Preparation Example 37 Preparation of Compound 34

[1317] Step 1: Preparation of 34-1

[1318] Reaction formula:

[1319]

[1320] Material ratio:

[1321] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195.06 1 eq 9.75g 50 1,2-Epoxydodecane 184.32 1 eq 9.2g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1322] Operation process:

[1323] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1324] Step 2: Preparation of 34-2

[1325] Reaction formula:

[1326]

[1327] Material ratio:

[1328] Material Name Molecular weight Feed ratio Feeding amount mmol 34-1 379.38 1eq 8g 21 N-nonanoic acid 158.24 1.1eq 3.67g 23 EDCI 192 2eq 8g 42 DMAP 122 0.2 eq 510mg 4.2 DCM - 20V 160mL -

[1329] Operation process:

[1330] 34-1, nonanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5°C to 0°C for 16 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 200 mL of saturated sodium bicarbonate solution and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 10 g of an oil.

[1331] Step 3: Preparation of compound 34

[1332] Reaction formula:

[1333]

[1334] Material ratio:

[1335] Material Name Molecular weight Feed ratio Feeding amount mmol 34-2 519.61 1eq 4g 7.7 cis-4-aminocyclohexanol 115.17 0.4 eq 355mg 3.1 potassium carbonate 138 2 eq 2.1g 15.4 Potassium iodide 166 1eq 1.28g 7.7 Acetonitrile - 20V 80mL -

[1336] Operation process:

[1337] 34-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.2 g of an oil.

[1338] 1 H 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).

[1339] MS (ES+) m / z): 992.7 (M) + .

[1340] Preparation Example 38 Preparation of Compound 36

[1341] Step 1: Synthesis of 1-[(6-{[6-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl][(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxy]dodec-2-yl 7-methyloctyl ester

[1342] Reaction formula:

[1343]

[1344] Material ratio:

[1345]

[1346] Operation process:

[1347] 1-[(6-bromohexanoyl)oxy]undec-2-yl octyl ester (1.30 g, 2.50 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-[(6-{[6-({2-[(7-methyloctanoyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl][(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxy]dodec-2-yl 7-methyloctyl ester (750 mg, 30.1% yield) as a yellow liquid.

[1348] 1H NMR(400MHz,CHLOROFORM-d)δ=5.13-5.03(m,2H),4.29-4.21(m,2H),4.10(brs,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.

[1349] LCMS:RT=2.777,m / z 993.5[M+H] + .

[1350] Preparation Example 39 Preparation of Compound 37

[1351] Step 1: Preparation of 37-1

[1352] Reaction formula:

[1353]

[1354] Material ratio:

[1355] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195.06 1 eq 9.75g 50 1,2-Epoxydodecane 184.32 1 eq 9.2g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1356] Operation process:

[1357] 6-Bromohexanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1358] Step 2: Preparation of 37-2

[1359] Reaction formula:

[1360]

[1361] Material ratio:

[1362] Material Name Molecular weight Feed ratio Feeding amount mmol 37-1 379.38 1eq 8g 21 n-Hexanoic acid 116.16 1.1eq 2.67g 23 EDCI 192 2eq 8g 42 DMAP 122 0.2 eq 510mg 4.2 DCM - 20V 160mL -

[1363] Operation process:

[1364] 37-1, n-hexanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5°C-0°C for 16 h. The product had an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 200 mL of saturated aqueous sodium bicarbonate and once with 200 mL of saturated aqueous sodium chloride. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 9 g of an oil.

[1365] Step 3: Preparation of compound 37

[1366] Reaction formula:

[1367]

[1368] Material ratio:

[1369] Material Name Molecular weight Feed ratio Feeding amount mmol 37-2 477.52 1eq 4g 8.4 cis-4-aminocyclohexanol 115.17 0.4 eq 386mg 3.35 potassium carbonate 138 2 eq 2.3g 16.8 Potassium iodide 166 1 eq 1.4g 8.4 Acetonitrile - 20V 80mL -

[1370] Operation process:

[1371] 37-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.2 g of an oil.

[1372] 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).

[1373] MS (ES+) m / z): 908.6 (M) + .

[1374] Preparation Example 40 Preparation of Compound 38

[1375] Step 1: Synthesis of 1-({6-[(6-{[2-(octanoyloxy)undecyl]oxy}-6-oxyylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexanoyl}oxy)undec-2-yloctyl ester

[1376] Reaction formula:

[1377]

[1378] Material ratio:

[1379] Material Name Molecular weight Feed ratio Feeding amount mmol 1-[(6-bromohexanoyl)oxy]undec-2-yloctyl ester 491 1.0 eq 1.00g 2.03 (1s,4s)-4-Aminocyclohexan-1-ol 115 0.45 eq 105mg 0.915 potassium carbonate 138 3.0 eq 843mg 6.10 Potassium iodide 166 1.5 eq 506mg 3.05 Acetonitrile - - 7mL - Tetrahydrofuran - - 3mL -

[1380] Operation process:

[1381] 1-[(6-bromohexanoyl)oxy]undec-2-yl octyl ester (1.00 g, 2.03 mmol) was dissolved in acetonitrile (7 mL), and (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({6-[(6-{[2-(octanoyloxy)undecyl]oxy}-6-oxyylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]hexanoyl}oxy)undec-2-yl octyl ester (540 mg, 28.3% yield) as a yellow liquid.

[1382] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.14-5.02(m,2H),4.29-4.20(m,2H),4.11(brs,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.

[1383] LCMS:RT=2.583,m / z 937.5[M+H] + .

[1384] Preparation Example 41 Preparation of Compound 39

[1385] Step 1: Preparation of 39-1

[1386] Reaction formula:

[1387]

[1388] Material ratio:

[1389] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195.06 1 eq 9.75g 50 1,2-Epoxydecane 156.27 1 eq 7.8g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1390] Operation process:

[1391] 6-Bromohexanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[1392] Step 2: Preparation of 39-2

[1393] Reaction formula:

[1394]

[1395] Material ratio:

[1396] Material Name Molecular weight Feed ratio Feeding amount mmol 39-1 351.33 1eq 7g 20 n-Decanic acid 172.27 1.1 eq 3.8g 22 EDCI 192 2 eq 7.7g 40 DMAP 122 0.2 eq 490mg 4 DCM - 20V 140mL -

[1397] Operation process:

[1398] To the reaction flask, 39-1, n-decanoic acid, EDCI, DMAP, and DCM were added and stirred at -5°C-0°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 150 mL of saturated aqueous sodium bicarbonate and once with 150 mL of saturated aqueous sodium chloride. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[1399] Step 3: Preparation of compound 39

[1400] Reaction formula:

[1401]

[1402] Material ratio:

[1403] Material Name Molecular weight Feed ratio Feeding amount mmol 39-2 505.58 1eq 4g 7.9 cis-4-aminocyclohexanol 115.17 0.4 eq 364mg 3.16 potassium carbonate 138 2 eq 2.2g 15.8 Potassium iodide 166 1 eq 1.3g 7.9 Acetonitrile - 20V 80mL -

[1404] Operation process:

[1405] 39-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2 g of an oil.

[1406] 1 H NMR(400MHz,Chloroform-d)δ5.07(qd,J=6.8,3.3Hz,2H),4.22(dd,J=11.8,3.3Hz,2H),4.01(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).

[1407] MS (ES+) m / z): 964.7 (M) + .

[1408] Preparation Example 42 Preparation of Compound 40

[1409] Step 1: Preparation of 40-1

[1410] Reaction formula:

[1411]

[1412] Material ratio:

[1413] Material Name Molecular weight Feed ratio Feeding amount mmol 6-Bromohexanoic acid 195.06 1 eq 9.75g 50 1,2-Epoxydecane 156.27 1 eq 7.8g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1414] Operation process:

[1415] 6-Bromohexanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[1416] Step 2: Preparation of 40-2

[1417] Reaction formula:

[1418]

[1419] Material ratio:

[1420] Material Name Molecular weight Feed ratio Feeding amount mmol 40-1 351.33 1eq 7g 20 n-heptanoic acid 130.19 1.1 eq 2.9g 22 EDCI 192 2 eq 7.7g 40 DMAP 122 0.2 eq 490mg 4 DCM - 20V 140mL -

[1421] Operation process:

[1422] 40-1, n-heptanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5°C-0°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction solution was washed once with 150 mL of saturated aqueous sodium bicarbonate and once with 150 mL of saturated aqueous sodium chloride. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[1423] Step 3: Preparation of compound 40

[1424] Reaction formula:

[1425]

[1426] Material ratio:

[1427] Material Name Molecular weight Feed ratio Feeding amount mmol 40-2 463.50 1eq 4g 8.6 cis-4-aminocyclohexanol 115.17 0.4 eq 400mg 3.45 potassium carbonate 138 2 eq 2.4g 17.2 Potassium iodide 166 1 eq 1.4g 8.6 Acetonitrile - 20V 80mL -

[1428] Operation process:

[1429] 40-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2 g of an oil.

[1430] 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).

[1431] MS (ES+) m / z): 880.6 (M) + .

[1432] Preparation Example 43 Preparation of Compound 41

[1433] Step 1: Synthesis of 1-({7-[(7-{[2-(octanoyloxy)undecyl]oxy}-7-oxyylideneheptyl)[(1s,4s)-4-hydroxycyclohexyl]amino]heptanoyl}oxy)undec-2-yloctyl ester

[1434] Reaction formula:

[1435]

[1436] Material ratio:

[1437]

[1438]

[1439] Operation process:

[1440] 1-[(7-bromoheptanoyl)oxy]undec-2-yloctyl ester (1.00 g, 1.98 mmol) was dissolved in acetonitrile (7 mL), and (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 1-({7-[(7-{[2-(octanoyloxy)undecyl]oxy}-7-oxyylideneheptyl)[(1s,4s)-4-hydroxycyclohexyl]amino]heptanoyl}oxy)undec-2-yloctyl ester (540 mg, 28.2% yield) as a yellow liquid.

[1441] 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.

[1442] LCMS:RT=2.625,m / z 965.6[M+H]+ .

[1443] Preparation Example 44 Preparation of Compound 42

[1444] Step 1: Preparation of 42-1

[1445] Reaction formula:

[1446]

[1447] Material ratio:

[1448] Material Name Molecular weight Feed ratio Feeding amount mmol 7-Bromoheptanoic acid 209.08 1 eq 10.5g 50 1,2-Epoxydecane 156.27 1 eq 7.8g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1449] Operation process:

[1450] 7-Bromoheptanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1451] Step 2: Preparation of 42-2

[1452] Reaction formula:

[1453]

[1454] Material ratio:

[1455]

[1456]

[1457] Operation process:

[1458] To the reaction flask, add 42-1, n-heptanoic acid, EDCI, DMAP, and DCM, and stir at -5°C-0°C for 16 h. The product exhibits an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture is washed once with 150 mL of saturated sodium bicarbonate solution and once with 150 mL of saturated sodium chloride solution. The organic phase is dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1459] Step 3: Preparation of compound 42

[1460] Reaction formula:

[1461]

[1462] Material ratio:

[1463] Material Name Molecular weight Feed ratio Feeding amount mmol 42-2 477.52 1eq 4.8g 10 cis-4-aminocyclohexanol 115.17 0.4 eq 460mg 4 potassium carbonate 138 2 eq 2.8g 20 Potassium iodide 166 1eq 1.66g 10 Acetonitrile - 20V 100mL -

[1464] Operation process:

[1465] 42-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.4 g of an oil.

[1466] 1 H 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).

[1467] MS (ES+) m / z): 908.7 (M) + .

[1468] Preparation Example 45 Preparation of Compound 43

[1469] Step 1: Preparation of 43-1

[1470] Reaction formula:

[1471]

[1472] Material ratio:

[1473]

[1474]

[1475] Operation process:

[1476] 8-Bromooctanoic acid, 1,2-epoxydodecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1477] Step 2: Preparation of 43-2

[1478] Reaction formula:

[1479]

[1480] Material ratio:

[1481] Material Name Molecular weight Feed ratio Feeding amount mmol 43-1 407.43 1eq 8g 19.6 N-nonanoic acid 158.24 1.1 eq 3.4g 21.6 EDCI 192 2 eq 7.5g 39.2 DMAP 122 0.2 eq 480mg 3.9 DCM - - 160mL -

[1482] Operation process:

[1483] Add 43-1, n-octanoic acid, EDCI, DMAP, and DCM to a reaction flask and stir at -5°C-0°C for 16 h. The product exhibits an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture is washed once with 200 mL of saturated sodium bicarbonate and once with 200 mL of saturated sodium chloride. The organic phase is dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 10 g of an oil.

[1484] Step 3: Preparation of compound 43

[1485] Reaction formula:

[1486]

[1487] Material ratio:

[1488] Material Name Molecular weight Feed ratio Feeding amount mmol 43-2 547.66 1eq 4g 7.3 cis-4-aminocyclohexanol 115.17 0.4 eq 336mg 2.9 potassium carbonate 138 2eq 2g 14.6 Potassium iodide 166 1 eq 1.2g 7.3 Acetonitrile - - 80mL -

[1489] Operation process:

[1490] 43-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 20:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.1 g of an oil.

[1491] 1H 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).

[1492] MS (ES+) m / z): 1048.8 (M) + .

[1493] Preparation Example 46 Preparation of Compound 44

[1494] Step 1: Synthesis of 2-(octanoyloxy)undecyl 8-[(8-{[2-(octanoyloxy)undecyl]oxy}-8-oxyylideneoctyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1495] Reaction formula:

[1496]

[1497] Material ratio:

[1498] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Octanoyloxy)undecyl 8-bromooctyl ester 519 1.0 eq 1.00g 1.92 (1s,4s)-4-Aminocyclohexan-1-ol 115 0.45eq 99.7mg 0.866 potassium carbonate 138 3.0 eq 797mg 5.77 Potassium iodide 166 1.5 eq 479mg 2.89 Acetonitrile - - 7mL - Tetrahydrofuran - - 3mL -

[1499] Operation process:

[1500] 2-(Octanoyloxy)undecyl 8-bromooctyl ester (1.00 g, 1.92 mmol) was dissolved in acetonitrile (7 mL). (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(Octanoyloxy)undecyl 8-[(8-{[2-(Octanoyloxy)undecyl]oxy}-8-oxyideneoctyl][(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (510 mg, 26.7% yield) as a yellow liquid.

[1501] 1H 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.

[1502] LCMS:RT=2.676,m / z 993.6[M+H] + .

[1503] Preparation Example 47 Preparation of Compound 45

[1504] Step 1: Preparation of 45-1

[1505] Reaction formula:

[1506]

[1507] Material ratio:

[1508] Material Name Molecular weight Feed ratio Feeding amount mmol 8-Bromooctanoic acid 223.11 1eq 11.15g 50 1,2-Epoxydecane 156.27 1 eq 7.8g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1509] Operation process:

[1510] 8-Bromooctanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1511] Step 2: Preparation of 45-2

[1512] Reaction formula:

[1513]

[1514] Material ratio:

[1515] Material Name Molecular weight Feed ratio Feeding amount mmol 45-1 379.38 1eq 8g 21 Octanoic acid 144.21 1.1 eq 3.3g 23 EDCI 192 2 eq 8.1g 42 DMAP 122 0.2 eq 510mg 4.2 DCM - 20V 160mL -

[1516] Operation process:

[1517] 45-1, n-octanoic acid, EDCI, DMAP, and DCM were added to the reaction flask and stirred at -5°C-0°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture was washed once with 150 mL of saturated sodium bicarbonate solution and once with 150 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1518] Step 3: Preparation of compound 45

[1519] Reaction formula:

[1520]

[1521] Material ratio:

[1522]

[1523]

[1524] Operation process:

[1525] 45-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.5 g of an oil.

[1526] 1 H 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).

[1527] MS (ES+) m / z): 964.7 (M) + .

[1528] Preparation Example 48 Preparation of Compound 46

[1529] Step 1: Preparation of 46-1

[1530] Reaction formula:

[1531]

[1532] Material ratio:

[1533] Material Name Molecular weight Feed ratio Feeding amount mmol 8-Bromooctanoic acid 223.11 1eq 11.15g 50 1,2-Epoxydecane 156.27 1 eq 7.8g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1534] Operation process:

[1535] 8-Bromooctanoic acid, 1,2-epoxydecane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1536] Step 2: Preparation of 46-2

[1537] Reaction formula:

[1538]

[1539] Material ratio:

[1540] Material Name Molecular weight Feed ratio Feeding amount mmol 46-1 379.38 1eq 8g 21 n-heptanoic acid 130.19 1.1eq 3g 23 EDCI 192 2 eq 8.1g 42 DMAP 122 0.2 eq 510mg 4.2 DCM - 20V 160mL -

[1541] Operation process:

[1542] To the reaction flask, add 46-1, n-heptanoic acid, EDCI, DMAP, and DCM, and stir at -5°C-0°C for 16 h. The product exhibits an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture is washed once with 150 mL of saturated aqueous sodium bicarbonate and once with 150 mL of saturated aqueous sodium chloride. The organic phase is dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 8 g of an oil.

[1543] Step 3: Preparation of compound 46

[1544] Reaction formula:

[1545]

[1546] Material ratio:

[1547] Material Name Molecular weight Feed ratio Feeding amount mmol 46-2 491.55 1 eq 4.92g 10 cis-4-aminocyclohexanol 115.17 0.4 eq 460mg 4 potassium carbonate 138 2 eq 2.8g 20 Potassium iodide 166 1eq 1.66g 10 Acetonitrile - 20V 100mL -

[1548] Operation process:

[1549] 46-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.5 g of an oil.

[1550] 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).

[1551] MS (ES+) m / z): 936.7 (M) + .

[1552] Preparation Example 49 Preparation of Compound 35

[1553] Step 1: Synthesis of ({[5-(pentane-3-oxy)pentyl]oxy}methyl)benzene

[1554] Reaction formula:

[1555]

[1556] Material ratio:

[1557] Material Name Molecular weight Feed ratio Feeding amount mmol 5-(Benzyloxy)pentan-1-ol 194 1.00eq 500mg 2.57 Pentan-3-one 86 1.50 eq 332mg 3.86 triethylsilane 116 1.50 eq 448mg 3.86 Ytterbium trifluoromethanesulfonate 620 0.01eq 15.9mg 0.0257 1,2-Dichloroethane - - 5.00mL -

[1558] Five parallel reactions, operation process:

[1559] 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 mixture was reacted at 92°C under nitrogen for 48 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. The crude product was purified by column chromatography to yield ({[5-(pentan-3-oxy)pentyl]oxy}methyl)benzene (1.25 g, 36.7% yield) as a colorless liquid.

[1560] Step 2: Synthesis of 5-(pentane-3-oxy)pentan-1-ol

[1561] Reaction formula:

[1562]

[1563] Material ratio:

[1564] Material Name Molecular weight Feed ratio Feeding amount mmol ({[5-(Pentan-3-oxy)pentyl]oxy}methyl)benzene 264 1.00 eq 1.25g 4.73 Palladium / Carbon 106 0.20eq 1.01g 0.945 hydrogen - (40Psi) - - Methanol - - 15.0mL -

[1565] Operation process:

[1566] Dissolve ({[5-(pentane-3-oxy)pentyl]oxy}methyl)benzene (1.25 g, 4.73 mmol) in methanol (15.0 mL). Add palladium / carbon (1.01 g, 945 μmol) under argon. Incubate at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. Completion of the reaction is monitored by TLC. The reaction mixture is filtered and concentrated. The mixture is then spin-dried to afford 5-(pentane-3-oxy)pentan-1-ol (732 mg, 88.8% yield) as a colorless liquid.

[1567] Step 3: Synthesis of 5-(pentane-3-oxy)pentanoic acid

[1568] Reaction formula:

[1569]

[1570] Material ratio:

[1571] Material Name Molecular weight Feed ratio Feeding amount mmol 5-(Pentan-3-oxy)pentan-1-ol 174 1.00eq 632mg 3.63 Diacetoxyiodobenzene 322 2.20eq 2.57g 7.98 2,2,6,6-Tetramethylpiperidinyloxide 157 0.20 eq 114mg 0.725 Acetonitrile - - 7.00mL - water - - 3.50mL -

[1572] Operation process:

[1573] 5-(Pentan-3-oxy)pentan-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-tetramethylpiperidinoxide (114 mg, 725 μmol). The mixture was allowed to react at 25°C under a nitrogen atmosphere for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was extracted with water and ethyl acetate, and concentrated. Column chromatography afforded 5-(pentan-3-oxy)pentanoic acid (538 mg, 70.8% yield) as a colorless liquid.

[1574] Step 4: Synthesis of 1-(benzyloxy)dodec-2-yl 5-(pentane-3-oxy)pentyl ester

[1575] Reaction formula:

[1576]

[1577] Material ratio:

[1578]

[1579] Operation process:

[1580] 1-(Benzyloxy)dodecan-2-ol (0.92 g, 3.15 mmol) and 5-(pentane-3-oxy)pentanoic acid (592 mg, 3.15 mmol) were dissolved in dichloromethane (10.0 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (904 mg, 4.72 mmol) and 4-dimethylaminopyridine (38.4 mg, 314 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodecan-3-yloctyl ester (1.40 g, 96.4% yield) as a colorless liquid.

[1581] Step 5: Synthesis of 1-hydroxydodec-2-yl 5-(pentane-3-oxy)pentyl ester

[1582] Reaction formula:

[1583]

[1584] Material ratio:

[1585]

[1586] Operation process:

[1587] 1-(Benzyloxy)dodec-2-yl 5-(pentane-3-oxy)pentyl ester (1.40 g, 3.03 mmol) was dissolved in methanol (20.0 mL). Palladium / carbon (643 mg, 605 μmol) was added under an argon atmosphere. The mixture was reacted at 35°C under a hydrogen atmosphere (40 psi) for 12 hours. The reaction was monitored by TLC. The reaction mixture was filtered and concentrated. 1-Hydroxydodec-2-yl 5-(pentane-3-oxy)pentyl ester (460 mg, 40.8% yield) was obtained as a colorless liquid by spin drying.

[1588] Step 6: Synthesis of 2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl 6-bromohexyl ester

[1589] Reaction formula:

[1590]

[1591] Material ratio:

[1592]

[1593]

[1594] Operation process:

[1595] 1-Hydroxydodec-2-yl 5-(pentane-3-oxy)pentyl ester (460 mg, 1.23 mmol) was dissolved in dichloromethane (10.0 mL). 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) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl 6-bromohexyl ester (619 mg, 91.2% yield) as a colorless liquid.

[1596] Step 7: Synthesis of 2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl 6-({6-oxyylidene-6-[(2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl)oxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester

[1597] Reaction formula:

[1598]

[1599] Material ratio:

[1600]

[1601] Operation process:

[1602] Dissolve 2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl 6-bromohexyl ester (519 mg, 944 μmol) in acetonitrile (6.00 mL), and add (1s,4s)-4-aminocyclohexan-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) in sequence. Finally, react at 80°C under nitrogen for 12 hours. The reaction is completed after monitoring by TLC. The mixture is filtered and concentrated. Purification by column chromatography gave a yellow liquid 2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl 6-({6-oxyylidene-6-[(2-{[5-(pentane-3-oxy)pentanoyl]oxy}dodecyl)oxy]hexyl}[(1s,4s)-4-hydroxycyclohexyl]amino)hexyl ester (342 mg, 34.1% yield).

[1603] 1H 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.

[1604] LCMS:RT=2.591,m / z 1053.5[M+H] + .

[1605] Preparation Example 50 Preparation of Compound 47

[1606] Step 1: Synthesis of 1-(benzyloxy)decan-3-ol

[1607] Reaction formula:

[1608]

[1609] Material ratio:

[1610] Material Name Molecular weight Feed ratio Feeding amount mmol 3-(Benzyloxy)propanal 164 1.0eq 2.0g 12.1 Bromo(heptyl)magnesium 203 1.1eq 2.73g 13.4 Tetrahydrofuran - - 20mL -

[1611] Operation process:

[1612] 3-(Benzyloxy)propanal (2.0 g, 12.1 mmol) was dissolved in tetrahydrofuran (20 mL). (Heptyl)magnesium bromide (2.73 g, 1.0 M, 13.4 mL, 13.4 mmol) was added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. After completion of the reaction, 50 mL of aqueous ammonium chloride was added, and the mixture was extracted with ethyl acetate twice, each time using 50 mL. The organic phase was concentrated and purified by column chromatography to afford 1-(benzyloxy)decan-3-ol (2.05 g, 63.6% yield), a colorless liquid.

[1613] Step 2: Synthesis of 1-(benzyloxy)decane-3-ylhexyl ester

[1614] Reaction formula:

[1615]

[1616] Material ratio:

[1617]

[1618] Operation process:

[1619] 1-(Benzyloxy)decan-3-ol (2.00 g, 7.56 mmol) and n-hexanoic acid (966 mg, 8.32 mmol) were dissolved in dichloromethane (20 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.18 g, 11.3 mmol) and 4-dimethylaminopyridine (184 mg, 1.51 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After addition of 10 mL of water, the mixture was extracted with dichloromethane twice, each time using 10 mL. The organic phase was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)decan-3-ylhexyl ester (2.00 g, 72.9% yield) as a colorless liquid.

[1620] Step 3: Synthesis of 1-hydroxydecane-3-ylhexyl ester

[1621] Reaction formula:

[1622]

[1623] Material ratio:

[1624] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)decan-3-ylhexyl ester 362 1.0 eq 2.00g 5.52 Palladium on carbon 105.9 0.2eq 1.17g 1.10 Methanol - - 20mL -

[1625] Operation process:

[1626] Dissolve 1-(Benzyloxy)decan-3-ylhexyl ester (2.00 g, 5.52 mmol) in methanol (20 mL), add palladium on carbon (1.17 g, 1.10 mmol), and react under hydrogen at 35°C, 40 psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1-hydroxydecan-3-ylhexyl ester (880 mg, 58.5% yield) as a colorless liquid.

[1627] Step 4: Synthesis of 3-(hexanoyloxy)decyl 8-bromooctyl ester

[1628] Reaction formula:

[1629]

[1630] Material ratio:

[1631]

[1632] Operation process:

[1633] 1-Hydroxydecane-3-yl hexyl ester (880 mg, 3.23 mmol) and 8-bromooctanoic acid (792 mg, 3.55 mmol) were dissolved in dichloromethane (10 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (928 mg, 4.85 mmol) and 4-dimethylaminopyridine (78.9 mg, 0.646 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. 10 mL of water was added, and the mixture was extracted with dichloromethane twice, each time using 10 mL. The organic phase was concentrated. The crude product was purified by column chromatography to give 3-(hexanoyloxy)decyl 8-bromooctyl ester (1.30 g, 84.2% yield) as a bright yellow liquid.

[1634] Step 5: Synthesis of 3-(hexanoyloxy)decyl 8-[(8-{[3-(hexanoyloxy)decyl]oxy}-8-oxyylideneoctyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1635] Reaction formula:

[1636]

[1637] Material ratio:

[1638] Material Name Molecular weight Feed ratio Feeding amount mmol 3-(Hexanoyloxy)decyl 8-bromooctyl ester 477 1.0 eq 1.20g 2.51 (1s,4s)-4-Aminocyclohexan-1-ol 115 0.45 eq 130mg 1.13 potassium carbonate 138 3.0eq 1.04g 7.54 Potassium iodide 166 1.5 eq 625mg 3.77 Acetonitrile - - 10mL - Tetrahydrofuran - - 3mL -

[1639] Operation process:

[1640] 3-(Hexanoyloxy)decyl 8-bromooctyl ester (1.20 g, 2.51 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 3-(hexanoyloxy)decyl 8-[(8-{[3-(hexanoyloxy)decyl]oxy}-8-oxyideneoctyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (680 mg, 29.8% yield) as a bright yellow liquid.

[1641] 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.

[1642] LCMS:RT=2.410,m / z 909.3[M+H] + .

[1643] Preparation Example 51 Preparation of Compound 48

[1644] Step 1: Synthesis of 1-(benzyloxy)nonan-2-ol

[1645] Reaction formula:

[1646]

[1647] Material ratio:

[1648] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Benzyloxy)acetaldehyde 150 1.0eq 4.0g 26.6 Bromo(heptyl)magnesium 203 1.1eq 5.96g 29.3 Tetrahydrofuran - - 40mL -

[1649] Operation process:

[1650] 3-(Benzyloxy)propanal (4.0 g, 26.6 mmol) was dissolved in tetrahydrofuran (40 mL). (Heptyl)magnesium bromide (5.96 g, 1.0 M, 29.3 mL, 29.3 mmol) was added at 0°C. The mixture was allowed to react at 25°C under nitrogen for 12 hours. After completion of the reaction, 50 mL of aqueous ammonium chloride was added, and the mixture was extracted with ethyl acetate twice, each time using 50 mL. The organic phase was concentrated and purified by column chromatography to afford 1-(benzyloxy)nonan-2-ol (5.62 g, 84.2% yield) as a colorless liquid.

[1651] Step 2: Synthesis of 1-(benzyloxy)nonan-2-ylhexyl ester

[1652] Reaction formula:

[1653]

[1654] Material ratio:

[1655]

[1656] Operation process:

[1657] 1-(Benzyloxy)nonan-2-ol (2.00 g, 7.99 mmol) and n-hexanoic acid (1.02 g, 8.79 mmol) were dissolved in dichloromethane (20 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (2.30 g, 11.9 mmol) and 4-dimethylaminopyridine (195 mg, 1.60 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After addition of 10 mL of water, the mixture was extracted with dichloromethane twice, each time using 10 mL. The organic phase was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)nonan-2-ylhexyl ester (2.60 g, 93.3% yield) as a colorless liquid.

[1658] Step 3: Synthesis of 1-hydroxynonan-2-ylhexyl ester

[1659] Reaction formula:

[1660]

[1661] Material ratio:

[1662] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)nonan-2-ylhexyl ester 348 1.0 eq 2.60g 7.46 Palladium on carbon 105.9 0.2eq 1.59g 1.49 Methanol - - 30mL -

[1663] Operation process:

[1664] Dissolve 1-(Benzyloxy)nonan-2-ylhexyl ester (2.60 g, 7.46 mmol) in methanol (30 mL), add palladium on carbon (1.59 g, 1.49 mmol), and react under hydrogen at 35°C, 40 Psi for 12 hours. Monitor the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain 1-hydroxynonan-2-ylhexyl ester (1.50 g, 77.8% yield) as a colorless liquid.

[1665] Step 4: Synthesis of 2-(hexanoyloxy)nonyl 8-bromooctyl ester

[1666] Reaction formula:

[1667]

[1668] Material ratio:

[1669]

[1670] Operation process:

[1671] 1-Hydroxynonan-2-yl hexyl ester (1.50 g, 5.81 mmol) and 8-bromooctanoic acid (1.55 g, 6.97 mmol) were dissolved in dichloromethane (15 mL). 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.67 g, 8.71 mmol) and 4-dimethylaminopyridine (141 mg, 1.16 mmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. TLC monitoring indicated the formation of new spots. After addition of 10 mL of water, the mixture was extracted with dichloromethane twice, each time using 10 mL. The organic phase was concentrated. The crude product was purified by column chromatography to yield 2-(hexanoyloxy)nonyl 8-bromooctyl ester (2.43 g, 90.3% yield) as a bright yellow liquid.

[1672] Step 5: Synthesis of 2-(hexanoyloxy)nonyl 8-[(8-{[2-(hexanoyloxy)nonyl]oxy}-8-oxyylideneoctyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1673] Reaction formula:

[1674]

[1675] Material ratio:

[1676] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Hexanoyloxy)nonyl 8-bromooctyl ester 462 1.0 eq 1.20g 2.59 (1s,4s)-4-Aminocyclohexan-1-ol 115 0.45 eq 134mg 1.17 potassium carbonate 138 3.0eq 1.07g 7.77 Potassium iodide 166 1.5 eq 644mg 3.88 Acetonitrile - - 10mL - Tetrahydrofuran - - 3mL -

[1677] Operation process:

[1678] 2-(Hexanoyloxy)nonyl 8-bromooctyl ester (1.20 g, 2.59 mmol) was dissolved in acetonitrile (10 mL), and (1s,4s)-4-aminocyclohexan-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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(hexanoyloxy)nonyl 8-[(8-{[2-(hexanoyloxy)nonyl]oxy}-8-oxyideneoctyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (750 mg, 32.8% yield) as a bright yellow liquid.

[1679] 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(brd,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.

[1680] LCMS:RT=2.328,m / z 881.3[M+H] + .

[1681] Preparation Example 52 Preparation of Compound 49

[1682] Step 1: Preparation of 49-1

[1683] Reaction formula:

[1684]

[1685] Material ratio:

[1686] Material Name Molecular weight Feed ratio Feeding amount mmol 8-Bromooctanoic acid 223.11 1eq 11.15g 50 1,2-Epoxyoctane 128.22 1 eq 6.4g 50 Ferric chloride 162 0.1 eq 810mg 5 Pyridine 79 0.1 eq 400mg 5

[1687] Operation process:

[1688] 8-Bromooctanoic acid, 1,2-epoxyoctane, ferric chloride, and pyridine were added to a reaction flask and allowed to react at room temperature for 16 hours. The product exhibited an Rf value of 0.4 according to TLC (PE:EA = 4:1). The reaction mixture was diluted with 200 mL of ethyl acetate, washed once with 200 mL of saturated sodium bicarbonate solution, and once with 200 mL of saturated sodium chloride solution. The organic phase was dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[1689] Step 2: Preparation of 49-2

[1690] Reaction formula:

[1691]

[1692] Material ratio:

[1693] Material Name Molecular weight Feed ratio Feeding amount mmol 49-1 351.33 1eq 7g 20 n-Valeric acid 102.13 1.1eq 2.24g 22 EDCI 192 2 eq 7.7g 40 DMAP 122 0.2 eq 490mg 4 DCM - 20V 140mL -

[1694] Operation process:

[1695] Add 49-1, n-valeric acid, EDCI, DMAP, and DCM to the reaction flask and stir at -5°C-0°C for 16 h. The product exhibits an Rf value of 0.6 according to TLC (PE:EA = 10:1). The reaction mixture is washed once with 150 mL of saturated sodium bicarbonate solution and once with 150 mL of saturated sodium chloride solution. The organic phase is dried over anhydrous magnesium sulfate, filtered, concentrated, and purified by column chromatography to yield 7 g of an oil.

[1696] Step 3: Preparation of compound 49

[1697] Reaction formula:

[1698]

[1699] Material ratio:

[1700] Material Name Molecular weight Feed ratio Feeding amount mmol 49-2 435.44 1eq 4g 9.2 cis-4-aminocyclohexanol 115.17 0.4 eq 423mg 3.67 potassium carbonate 138 2 eq 2.5g 18.4 Potassium iodide 166 1 eq 1.5g 9.2 Acetonitrile - 20V 80mL -

[1701] Operation process:

[1702] 49-2, cis-4-aminocyclohexanol, potassium carbonate, potassium iodide, and acetonitrile were added to a reaction flask and reacted at 75°C for 16 h. The product exhibited an Rf value of 0.6 according to TLC (DCM:MeOH = 10:1). The reaction mixture was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to yield 2.2 g of an oil.

[1703] 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).

[1704] MS (ES+) m / z): 824.6 (M) + .

[1705] Preparation Example 53 Preparation of Compound 50

[1706] Step 1: Synthesis of 1-[(6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxy]undec-2-yloctyl ester

[1707] Reaction formula:

[1708]

[1709] Material ratio:

[1710] Material Name Molecular weight Feed ratio Feeding amount mmol 1-[(6-bromohexanoyl)oxy]undec-2-yloctyl ester 491 1.0eq 1.0g 2.03 (1s,4s)-4-Aminocyclohexan-1-ol 115 10.0eq 2.34g 20.3 Acetonitrile - - 10mL -

[1711] Operation process:

[1712] Dissolve 1-[(6-bromohexanoyl)oxy]undec-2-yl octyl ester (1.0 g, 2.03 mmol) in acetonitrile (10 mL), add (1s,4s)-4-aminocyclohexan-1-ol (2.34 g, 20.3 mmol), and react at 40°C under nitrogen for 24 hours. Monitor the reaction by TLC, concentrate, and purify by column chromatography to yield 1-[(6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxy]undec-2-yl octyl ester (870 mg, 81.3% yield), a yellow liquid.

[1713] Step 2: Synthesis of 2-(hexanoyloxy)nonyl 8-[(6-{[2-(octanoyloxy)undecyl]oxy}-6-oxyylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester

[1714] Reaction formula:

[1715]

[1716] Material ratio:

[1717]

[1718] Operation process:

[1719] 2-(Hexanoyloxy)nonyl 8-bromooctyl ester (746 mg, 1.61 mmol) was dissolved in acetonitrile (10 mL). 1-[(6-{[(1s,4s)-4-hydroxycyclohexyl]amino}hexanoyl)oxy]undec-2-yl octyl 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 mixture was reacted at 80°C under nitrogen for 12 hours. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave 2-(hexanoyloxy)nonyl 8-[(6-{[2-(octanoyloxy)undecyl]oxy}-6-oxyylidenehexyl)[(1s,4s)-4-hydroxycyclohexyl]amino]octyl ester (850 mg, 63.8% yield) as a bright yellow liquid.

[1720] 1H NMR(400MHz,CHLOROFORM-d)δ=5.15-5.01(m,2H),4.28-4.21(m,2H),4.10(brs,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.

[1721] LCMS:RT=2.456,m / z 908.8[M+H] + .

[1722] Preparation Example 54 Preparation of Compound 51

[1723] Step 1: Synthesis of 4-[3-(benzyloxy)propoxy]butan-1-ol

[1724] Reaction formula:

[1725]

[1726] Material ratio:

[1727] Material Name Molecular weight Feed ratio Feeding amount mmol [(3-Bromopropoxy)methyl]benzene 229 1.00 eq 3.00g 13.0 1,4-Butanediol 90 5.00eq 5.90g 65.4 potassium hydroxide 56 7.50eq 5.51g 98.2 dimethyl sulfoxide - - 30.0mL -

[1728] Operation process:

[1729] Dissolve [(3-bromopropoxy)methyl]benzene (3.00 g, 13.0 mmol) and 1,4-butanediol (5.90 g, 65.4 mmol) in dimethyl sulfoxide (30.0 mL). Add potassium hydroxide (5.51 g, 98.2 mmol). Incubate at 25°C under nitrogen for 1 hour. Completion is monitored by TLC. The reaction mixture is washed with water and extracted. Column chromatography yields the crude product as a colorless liquid, 4-[3-(benzyloxy)propoxy]butan-1-ol (1.97 g, 63.1% yield).

[1730] Step 2: Synthesis of 4-[3-(benzyloxy)propoxy]butyric acid

[1731] Reaction formula:

[1732]

[1733] Material ratio:

[1734] Material Name Molecular weight Feed ratio Feeding amount mmol 4-[3-(Benzyloxy)propoxy]butan-1-ol 238 1.00 eq 1.00g 4.20 Diacetoxyiodobenzene 322 2.20eq 2.97g 9.23 2,2,6,6-Tetramethylpiperidinyloxide 157 0.20 eq 131mg 0.839 Acetonitrile - - 10.0mL - water - - 5.00mL -

[1735] Operation process:

[1736] 4-[3-(Benzyloxy)propoxy]butan-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-tetramethylpiperidinyloxide (131 mg, 839 μmol). The mixture was allowed to react at 25°C under a nitrogen atmosphere for 12 hours. Completion of the reaction was monitored by TLC. The reaction solution was extracted with water and ethyl acetate, and concentrated. Column chromatography afforded 4-[3-(Benzyloxy)propoxy]butanoic acid (918 mg, 86.7% yield) as a colorless liquid.

[1737] Step 3: Synthesis of 1-(benzyloxy)dodec-2-yl 5-(pentane-3-oxy)pentyl ester

[1738] Reaction formula:

[1739]

[1740] Material ratio:

[1741]

[1742] Operation process:

[1743] 1-Hydroxydecane-2-yl heptyl ester (324 mg, 1.13 mmol) and 4-[3-(benzyloxy)propoxy]butanoic acid (285 mg, 1.13 mmol) were dissolved in dichloromethane (5.00 mL). 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (325 mg, 1.70 mmol) and 4-dimethylaminopyridine (13.8 mg, 113 μmol) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. The reaction was monitored by TLC and the reaction solution was concentrated. The crude product was purified by column chromatography to yield 1-(benzyloxy)dodec-2-yl 5-(pentane-3-oxy)pentyl ester (531 mg, 90.1% yield) as a colorless liquid.

[1744] Step 4: Synthesis of 1-{[4-(3-hydroxypropoxy)butyryl]oxy}decan-2-ylheptyl ester

[1745] Reaction formula:

[1746]

[1747] Material ratio:

[1748]

[1749] Operation process:

[1750] 1-(Benzyloxy)dodec-2-yl 5-(pentane-3-oxy)pentyl ester (531 mg, 1.02 mmol) was dissolved in methanol (6.00 mL). Palladium / carbon (217 mg, 203 μmol) was added under an argon atmosphere. The mixture was reacted at 35°C under a hydrogen atmosphere (35 psi) for 12 hours. The reaction was monitored by TLC. The reaction mixture was filtered and concentrated. The mixture was dried to give a colorless liquid, 1-{[4-(3-hydroxypropyloxy)butyryl]oxy}decan-2-ylheptyl ester (439 mg, 99.9% yield).

[1751] Step 5: Synthesis of 1-{[4-(3-bromopropoxy)butyryl]oxy}decan-2-ylheptyl ester

[1752] Reaction formula:

[1753]

[1754] Material ratio:

[1755]

[1756] Operation process:

[1757] 1-{[4-(3-hydroxypropoxy)butyryl]oxy}decan-2-ylheptyl ester (389 mg, 903 μmol) was dissolved in dichloromethane (10.0 mL). Triphenylphosphine (473 mg, 1.81 mmol) and carbon tetrabromide (599 mg, 1.81 mmol) were added at 0°C. The mixture was reacted at 25°C under a nitrogen atmosphere for 12 hours. The reaction was monitored by TLC. The mixture was concentrated and purified by column chromatography to obtain a colorless liquid 1-{[4-(3-bromopropoxy)butyryl]oxy}decan-2-ylheptyl ester (400 mg, 89.7% yield).

[1758] Step 6: Synthesis of 1-{[4-(3-{[3-(4-{[2-(heptanoyloxy)decyl]oxy}-4-oxyylidenebutyloxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)butyryl]oxy}decan-2-ylheptyl ester

[1759] Reaction formula:

[1760]

[1761] Material ratio:

[1762]

[1763] Operation process:

[1764] 1-{[4-(3-bromopropoxy)butyryl]oxy}decan-2-ylheptyl ester (300 mg, 607 μmol) was dissolved in acetonitrile (4.00 mL), and (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) were added in sequence. The mixture was reacted at 80 ° C under nitrogen protection for 12 hours. The reaction was completed after monitoring by TLC. The mixture was filtered and concentrated. Purification by column chromatography gave yellow liquid 1-{[4-(3-{[3-(4-{[2-(heptanoyloxy)decyl]oxy}-4-oxobutylenebutoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)butanoyl]oxy}decan-2-ylheptyl ester (163 mg, 27.8% yield).

[1765] 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.

[1766] LCMS:RT=2.439,m / z 941.4[M+H] + .

[1767] Preparation Example 55 Preparation of Compound 52

[1768] Step 1: Synthesis of 1-{[2-(3-bromopropoxy)acetyl]oxy}undec-2-yloctyl ester

[1769] Reaction formula:

[1770]

[1771] Material ratio:

[1772]

[1773]

[1774] Operation process:

[1775] Dissolve 1-hydroxyundec-2-yloctyl ester (450 mg, 1.43 mmol) in dichloromethane (5.00 mL). 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) were added sequentially. The mixture was reacted at 25°C under nitrogen for 12 hours. Completion of the reaction was monitored by TLC, and the reaction solution was concentrated. Column chromatography of the crude product afforded 1-{[2-(3-bromopropoxy)acetyl]oxy}undec-2-yloctyl ester (675 mg, 95.5% yield) as a colorless liquid.

[1776] Step 2: Synthesis of 1-{[2-(3-{[3-(2-{[2-(octanoyloxy)undecyl]oxy}-2-oxyylideneethoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)acetyl]oxy}undec-2-yloctyl ester

[1777] Reaction formula:

[1778]

[1779] Material ratio:

[1780]

[1781] Operation process:

[1782] 1-{[2-(3-bromopropoxy)acetyl]oxy}undecan-2-yloctyl ester (575 mg, 1.17 mmol) was dissolved in acetonitrile (6.00 mL), and (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) were added in sequence. Finally, the mixture was reacted at 80°C under nitrogen protection for 12 hours. The reaction was completed after monitoring by TLC. The mixture was filtered and concentrated. Purification by column chromatography gave yellow liquid 1-{[2-(3-{[3-(2-{[2-(octanoyloxy)undecyl]oxy}-2-oxoylideneethoxy)propyl][(1s,4s)-4-hydroxycyclohexyl]amino}propoxy)acetyl]oxy}undec-2-yloctyl ester (157 mg, 14.3% yield).

[1783] 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.

[1784] LCMS:RT=2.564,m / z 941.3[M+H] + .

[1785] Example 1 Preparation and Detection of Lipid Nanoparticles (LNP)

[1786] To verify whether the lipid nanoparticle (LNP) preparation prepared by the ionizable lipid compound disclosed in the present application can effectively encapsulate mRNA and maintain the structural integrity of mRNA. The prepared ionizable lipid compound, distearoylphosphatidylcholine (DSPC, purchased from Nippon Seika Co., Ltd., Catalog No.: S01005), cholesterol (purchased from Nippon Seika Co., Ltd., Catalog No.: O01001) and dimyristoylglycerol-polyethylene glycol 2000 (DMG-PEG2000, purchased from Guobang Pharmaceutical, Catalog No.: O02005) were dissolved in ethanol (manufacturer: Nanjing Chemical Reagent Co., Ltd., purity 99.6%) solution, and then mixed according to a certain molar ratio to prepare a mixed lipid ethanol solution, wherein the total lipid concentration was 12.5 mM (the unit of measurement "M" in this application refers to mol / L). Firefly luciferase (Fluc) mRNA was diluted in 25 mM sodium acetate buffer at pH 5.0 to obtain an mRNA solution. By using a microfluidic device, the flow rate was controlled at 12 mL / min, the volume ratio of the mixed lipid ethanol solution to the mRNA solution prepared in the previous step was controlled to be 1:3, and lipid nanoparticles were prepared according to the nitrogen-phosphorus ratio of the ionizable lipid to the mRNA of 3 to 15:1. The ethanol was removed by dialysis against 20 mM Tris acetate for 12 to 24 hours. Finally, the LNP solution was filtered through a sterile filter with a pore size of 0.22 μm (manufacturer: Millex, product number: SLGPR33RB) and concentrated by ultrafiltration (manufacturer: Amicon-Ultra, molecular weight cutoff: 10 kDa) to obtain the LNP preparation obtained by encapsulating Fluc mRNA with the ionizable lipid described in this application and DSPC, cholesterol and DMG-PEG2000. The particle size and polydispersity index (PDI) of each LNP formulation were determined using dynamic light scattering using a Malvern Zetasizer Ultra instrument (Manufacturer: Malvern). The encapsulation efficiency of the LNPs was determined using the Quant-it Ribogreen RNA Quantification Kit (Manufacturer: ThermoFisher Scientific, Catalog No.: R11490).

[1787] Table 1

[1788]

[1789] In the art, a PDI of less than 0.3 indicates relatively uniform nanoparticle size within the LNP formulation. Encapsulation efficiency is used to assess whether the LNP can effectively encapsulate mRNA, with an encapsulation efficiency above 70% indicating effective mRNA encapsulation. The LNPs prepared in this application maintained a particle size of 60 to 80 nm, a PDI of less than 0.2, and an encapsulation efficiency exceeding 90%.

[1790] Example 2 In vivo animal studies of LNP preparations

[1791] In this example, the LNPs prepared in Example 1 were injected into 6- to 8-week-old female Balb / C mice (Weitonglihua) via tail vein or lower limb intramuscular injection at a dose of 5 μg / mouse (n=3, i.e., 3 mice were injected and tested per group, and the data presented are the mean values ​​for each group). D-luciferin potassium salt was injected intraperitoneally at specific time points after administration (4 hours, 24 hours, and 48 hours in this example). Luminescence was then detected using an IVIS Spectrum small animal in vivo imager (manufacturer: PerkinElmer). The total luminescence intensity of the expression sites in the mice (e.g., liver, lower limb administration sites, etc.) was calculated. The higher the luminescence intensity, the higher the luciferase expression, i.e., the better the expression of the corresponding LNP formulation in the mice. The total luminescence intensity was measured by bioluminescence imaging, and the luminescence intensity data of the luminescent sites were collected 6 to 15 minutes (min) after the intraperitoneal injection of D-luciferin potassium salt. The total luminescence intensity of the expression areas in the mice was calculated using Living Image software (manufacturer: PerkinElmer). Normally, the total luminescence intensity reading of mice not treated with drugs is on the order of 10 5 .

[1792] Referring to the above-mentioned in vivo mouse experimental method, the LNP preparation in Example 1 was injected into 6-8 week old female Balb / C mice via the tail vein at a dose of 5 μg / mouse. The total luminescence intensity of the liver area was counted. The test results are shown in Table 2 and Figure 1 The LNP preparations tested in this example all showed strong expression in mice, with an AUC range of 10 9 ~10 12 This indicates that the LNP preparations corresponding to the ionizable lipids described in the preparation examples can effectively deliver mRNA into the body and express it.

[1793] Table 2 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1794] Group Liver AUC (p / s*hour) Compound 1 3.94E+12 Compound 2 2.78E+12 Compound 3 2.89E+12 Compound 5 3.87E+12

[1795] Example 3 Preparation and Detection of Lipid Nanoparticles (LNP)

[1796] In this example, compound 4 and compound 14 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 3, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 3, the particle size of the LNP preparations prepared in this example was between 70 and 90 nm, the PDI was less than 0.25, and the encapsulation efficiency was greater than 90%.

[1797] Table 3

[1798]

[1799] Example 4 In vivo animal studies of LNP preparations

[1800] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 3 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the living mice was counted. The test results are shown in FIG. Figure 2 and Table 4. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1801] Table 4 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1802] Group Liver AUC (p / s*hour) Compound 4 2.65E+12 Compound 14 1.59E+12

[1803] Example 5 Preparation and Detection of Lipid Nanoparticles (LNP)

[1804] This example selects compound 6, compound 7, compound 10, compound 12, compound 13, compound 17, compound 19, compound 21 and compound 22 as ionizable lipids, and prepares LNP preparations (encapsulating Fluc mRNA) in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 5. The mRNA of this example is diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 is used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this example are measured. As shown in Table 5, the particle size of the LNP preparations prepared in this example is between 55 and 100 nm, the PDI is less than 0.2, and the encapsulation efficiency is higher than 85% (except compound 6).

[1805] Table 5

[1806]

[1807] Example 6 In vivo animal studies of LNP preparations

[1808] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 5 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 3 and Table 6. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1809] Table 6 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1810]

[1811]

[1812] Example 7 Preparation and Detection of Lipid Nanoparticles (LNP)

[1813] In this embodiment, compound 9, compound 20, compound 25, compound 26, compound 27, compound 28 and compound 30 were selected as ionizable lipids, and LNP preparations (encapsulated Fluc mRNA) were prepared in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 7. The mRNA of this embodiment was diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this embodiment were measured. As shown in Table 7, the particle size of the LNP preparations prepared in this embodiment was between 50 and 80 nm, the PDI was less than 0.15, and the encapsulation efficiency was higher than 90%.

[1814] Table 7

[1815]

[1816] Example 8 In vivo animal studies of LNP preparations

[1817] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 7 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 4 and Table 8. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1818] Table 8 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1819] Group Liver AUC (p / s*hour) Compound 9 6.51E+10 Compound 20 1.57E+12 Compound 25 1.87E+11 Compound 26 5.44E+11 Compound 27 9.18E+11 Compound 28 1.00E+12 Compound 30 1.59E+12

[1820] Example 9 Preparation and Detection of Lipid Nanoparticles (LNP)

[1821] In this embodiment, compound 8, compound 15, compound 23, compound 24 and compound 29 were selected as ionizable lipids, and LNP preparations (encapsulated Fluc mRNA) were prepared in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 9. The mRNA of this embodiment was diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this embodiment were measured. As shown in Table 9, the particle size of the LNP preparations prepared in this embodiment was between 50 and 90 nm, the PDI was less than 0.15, and the encapsulation efficiency was higher than 85%.

[1822]

[1823] Compound I-6-II was cited from international application WO2024017250A1 and Chinese application CN117417264A, and was synthesized using the same preparation method as WO2024017250A1.

[1824] Table 9

[1825]

[1826] Example 10 In vivo animal studies of LNP preparations

[1827] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 9 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 5 and Table 10. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1828] Table 10 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1829]

[1830]

[1831] Example 11 Preparation and Detection of Lipid Nanoparticles (LNP)

[1832] In this embodiment, compound 11, compound 16, compound 18, compound 31 and compound 32 were selected as ionizable lipids, and LNP preparations (encapsulated Fluc mRNA) were prepared in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 11. The mRNA of this embodiment was diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this embodiment were measured. As shown in Table 11, the particle size of the LNP preparations prepared in this embodiment was between 55 and 120 nm, the PDI was less than 0.2, and the encapsulation efficiency was higher than 85%.

[1833] Table 11

[1834]

[1835] Example 12 In vivo animal study of LNP preparations

[1836] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 11 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 6 and Table 12. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1837] Table 12 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1838] Group Liver AUC (p / s*hour) Compound 11 3.15E+10 Compound 16 7.39E+11 Compound 18 2.26E+12 Compound 31 1.79E+12 Compound 32 5.26E+11

[1839] Example 13 Preparation and Detection of Lipid Nanoparticles (LNP)

[1840] In this example, Compound 53 and Compound 54 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared according to the molar ratios and nitrogen-phosphorus ratios in Table 13, with reference to Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI, and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 13, the particle size of the LNP preparations prepared in this example was between 130 and 140 nm, the PDI was less than 0.55, and the encapsulation efficiency was greater than 85%.

[1841] Table 13

[1842]

[1843] Example 14 In vivo animal study of LNP preparations

[1844] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 13 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 7 and Table 14. As can be seen, the LNP preparations prepared in this example were strongly expressed in mice.

[1845] Table 14 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1846] Group Liver AUC (p / s*hour) Compound 53 1.82E+11 Compound 54 6.68E+11

[1847] Example 15 Preparation and Detection of Lipid Nanoparticles (LNP)

[1848] In this embodiment, compound 33, compound 34, compound 36, compound 37, compound 38, compound 39, compound 40, compound 41, compound 42, compound 43, compound 44, compound 45 and compound 46 were selected as ionizable lipids, and LNP preparations (encapsulating Fluc mRNA) were prepared in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 15. The mRNA of this embodiment was diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this embodiment were measured. As shown in Table 15, the particle size of the LNP preparations prepared in this embodiment was between 50 and 90 nm, the PDI was less than 0.15, and the encapsulation efficiency was higher than 85%.

[1849] Table 15

[1850]

[1851]

[1852] Example 16 In vivo animal study of LNP preparations

[1853] Referring to the in vivo test method of mice in Example 2, the LNP reagent prepared in Example 16 was injected into female Balb / C mice aged 6 to 8 weeks through the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 8 and Table 16. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1854] Table 16 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1855] Group Liver AUC (p / s*hour) Compound 33 1.18E+12 Compound 34 1.72E+12 Compound 36 2.12E+12 Compound 37 1.13E+12 Compound 38 2.48E+12 Compound 39 2.12E+12 Compound 40 1.43e+12 Compound 41 2.27E+12 Compound 43 3.68E+12 Compound 44 2.00E+12 Compound 45 2.56E+12 Compound 46 2.21E+12

[1856] Example 17 Preparation and Detection of Lipid Nanoparticles (LNP)

[1857] In this embodiment, compound 35, compound 47, compound 48, compound 49, compound 50, compound 51 and compound 52 were selected as ionizable lipids, and LNP preparations (encapsulated Fluc mRNA) were prepared in the manner of reference example 1 according to the molar ratio and nitrogen-phosphorus ratio in Table 17. The mRNA of this embodiment was diluted in a 25mM sodium acetate solution with a pH of 5.0, and a 20mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this embodiment were measured. As shown in Table 17, the particle size of the LNP preparations prepared in this embodiment was between 45 and 100 nm, the PDI was less than 0.25, and the encapsulation efficiency was higher than 90%.

[1858] Table 17

[1859]

[1860] Example 18 In vivo animal study of LNP preparations

[1861] Referring to the in vivo test method of Example 2, the LNP reagent prepared in Example 17 was injected into female Balb / C mice aged 6 to 8 weeks via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the liver of the mice was counted. The test results are shown in FIG. Figure 9 and Table 18. It can be seen that the LNP preparations prepared in this example were strongly expressed in mice.

[1862] Table 18 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[1863] Group Liver AUC (p / s*hour) Compound 35 9.38E+11 Compound 47 1.09E+12 Compound 48 1.15E+12 Compound 50 3.33E+12 Compound 51 6.13E+11 Compound 52 2.07E+10

[1864] Example 19 Preparation and Detection of Lipid Nanoparticles (LNP)

[1865] In this example, compound 1, compound 5, compound 7, compound 10, compound 17 and compound 21 were selected as ionizable lipids, and LNP preparations (encapsulating RSV mRNA, Uniprot: accession No. P03420) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 19, with reference to Example 1. The mRNA of this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this example were measured. As shown in Table 19, the particle size of the LNP preparations prepared in this example was between 60 and 90 nm, the PDI was less than 0.15, and the encapsulation efficiency was higher than 90%.

[1866] Table 19

[1867]

[1868] Example 20 In vivo animal study of LNP preparations

[1869] Referring to the in vivo test method of mice in Example 2, the RSV-LNP reagent prepared in Example 19 was injected into female BABL / c mice aged 6 to 8 weeks through the tail vein at a dose of 40 μg / mouse, and whole blood of the mice was collected 6 hours after injection. Serum was separated from the whole blood by centrifugation at 4°C and 2000g for 10 minutes and stored at -80°C for analysis. According to the manufacturer's instructions, the RSV antigen expression concentration in the mouse serum was quantitatively detected by enzyme-linked immunosorbent assay using the Novozymes Respiratory Syncytial Virus pre-F Elisa Kit (Φ&Ⅳ) (Cat. No.: DD3939). The test results are shown in the table. Figure 10 And Table 20 (RSV antigen concentration in serum (n=5), statistical analysis by ANOVA, ****p<0.0001, *****p<0.0001, compared with the placebo group). Compared with the placebo group, the RSV antigen concentration of animals in the Compound 1, Compound 5, Compound 7, Compound 10, Compound 17, and Compound 21 groups all increased significantly, and were significantly better than Compounds I-6-II.

[1870] Table 20 RSV antigen content in BABL / c mouse serum 6 hours after intravenous administration

[1871]

[1872]

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in, R 1 is a hydroxyl group, or a C substituted by 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 is -O-, -O-Z 1c -C(=O)- O- or chemical bonds; Z 1a 、Z 1b and Z 1c Independently C 1-6 alkylene; Z 2 for -O- or -OZ 2c -C(=O)-O-; Z 2a 、Z 2b and Z 2c Independently C 1-6 alkylene; W 1 and W 2 independently a chemical bond 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 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 is a 3-10 membered heterocycloalkyl group; the number of heteroatoms in the heterocycloalkyl group is 1-3, and the heteroatoms are independently N, O or S; R 3 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 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 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 is a 3-10 membered heterocycloalkyl group; the number of heteroatoms in the heterocycloalkyl group is 1-3, and the heteroatoms are independently N, O or S; R 5 H, C 1-15 alkyl, R 5a C 1-6 Alkylene; R 5b and R 5c Independently C 1-15 alkyl.

2. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein Z 1 for or chemical bonds; Z 1a and Z 1b Independently C 1-6 alkylene; Z 2 for Z 2a and Z 2b Independently C 1-6 alkylene; R 2a For chemical bonds or C 1-6 Alkylene; R 2b 、R 2c and R 2d Independently C 1-15 Alkyl or C 1-15 alkenyl; R 4c 、R 4d and R 4e Independently C 1-15 Alkyl or C 1-15 Alkenyl.

3. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 2, wherein It meets one or more of the following conditions: (1)R 1 is a hydroxyl group or a C substituted by a hydroxyl group 1-3 alkyl; (2) Ring A is C 4-6 cycloalkylene; (3)L 1 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 C 1-3 alkylene; (6)Z 1 for or chemical bond, * indicates the end connected to X; (7)Z 2a and Z 2b Independently C 1-4 alkylene; (8)Z 2 for * indicates the end connected to Y; (9)W 1 and W 2 independently a chemical bond or C 1-3 Alkylene, such as chemical bonds, methylene, (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 Preferably -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 H, C 5-15 Alkyl, -R 3a -OC 5-15 alkyl, (21)R 4a 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 H, C 5-15 alkyl, 4. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 3, wherein It meets one or more of the following conditions: (1)R 1 for Preferably (2) Ring A is (3) for Preferably, for More preferably for (4)L 1 is a chemical bond; (5) X and Y are independently (6)Z 1a and Z 1b for (7)Z 1 for or 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 C 1-3 Alkylene, such as methylene, (11)R 2a 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 is a methylene group; (17)R 3b is a methylene group; (18)R 3c for (19)R 3d for (20)R 3e for (21)R 3 For H, (twenty two) for (23)R 4b is a methylene group; (24)R 4c for (25)R 4d for (26)R 4e for (27)R 4 for (28)R 5a is a methylene group; (29)R 5b for (30)R 5c for (31)R 5 For H, (32) for 5. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein It meets one or more of the following conditions: (1) X and Y are independently (2)Z 1a and Z 1b are independently methylene or (3)Z 1c C 1-3 Alkylene, such as methylene, (4)Z 1 for -O- or *-OZ 1c -C(=O)-O-, * represents 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 Alkylene, such as methylene, (7)Z 2 for -O- or *-OZ 2c -C(=O)-O-, * represents the end connected to Y; Preferably, Z 2 for * indicates the end connected to Y; More preferably, Z 2 for * 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 is a 5-8 membered heterocycloalkyl group; in the heterocycloalkyl group, the number of heteroatoms is preferably 2, and the heteroatom is preferably S; Preferably, R 2ba for (10)R 2 for (11)R 3e C 5-15 Alkyl groups, e.g. (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 is a 5-8 membered heterocycloalkyl group; in the heterocycloalkyl group, the number of heteroatoms is preferably 2, and the heteroatom is 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 Alkenyl is independently C 2-15 Alkenyl.

6. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein The compound of formula I satisfies any of the following schemes: 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 as described in any one of claims 1 to 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 as described in any one of claims 1 to 5; in, Preferably W 1 and W 2 Preferably C 1-3 alkylene; More preferably, the compound of formula I has the following structure: Among them, R 1 , Ring A, X, Y, W 1 、W 2 、R 2b 、R 3 、R 4d and R 5 The definition as described in any one of claims 1 to 5; Among them, ring A is preferably Further 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 as described in any one of claims 1 to 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 as described in any one of claims 1 to 5; preferably, R 1 is a hydroxyl group, or a C substituted by 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 C 1-6 alkylene; W 1 and W 2 independently a chemical bond 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 as described in any one of claims 1 to 5; Preferably, R 1 is a hydroxyl group, or a C substituted by a hydroxyl group 1-3 alkyl; X and Y are independently C 3-10 alkylene; W 1 and W 2 Independently 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 as described in any one of claims 1 to 5; Preferably, R 1 is a hydroxyl group, or a C substituted by a hydroxyl group 1-3 alkyl; X and Y are independently C 3-10 alkylene; W 1 and W 2 Independently 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 is hydroxyl, or -CH2-OH; X and Y are independently C 4-8 Alkylene, preferably C 5-7 Alkylene, e.g. W 1 and W 2 Independently C 1-3 alkylene groups, such as methylene; R 2b C 6-10 Alkyl, preferably C 6-8 Alkyl groups, e.g. R 3 C 1-15 Alkyl, preferably C 8-12 Alkyl, more preferably C 8-10 Alkyl groups, e.g. R 4d C 1-15 Alkyl, preferably C 4-9 Alkyl, more preferably C 5-8 Alkyl groups, e.g. R 5 C 1-15 Alkyl, preferably C 6-12 Alkyl, more preferably C 7-10 Alkyl groups, e.g. 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 as described in any one of claims 1 to 5; Preferably, X and Y are independently C 5-7 Alkylene, e.g. W 1 and W 2 Independently C 1-3 alkylene groups, such as methylene; R 2b C 6-8 Alkyl groups, e.g. R 3 C 8-10 Alkyl groups, e.g. R 4d C 5-8 Alkyl groups, e.g. R 5 C 7-10 Alkyl groups, e.g. The compound of formula I described in scheme (6) is 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 to 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 is a hydroxyl group, or a C substituted by 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 independently a chemical bond 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 is 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 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 is 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 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 or a pharmaceutically acceptable salt thereof according to claim 1, wherein The compound of formula I is any of the following compounds:

8. A lipid carrier comprising a substance Z, wherein the substance Z is a compound of formula I according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof.

9. The lipid carrier according to claim 8, wherein It meets one or more of the following conditions: (1) The lipid carrier further comprises a diluent, such as phosphate buffer or Tris buffer; (2) The lipid carrier further comprises a phospholipid, which is a phospholipid molecule having an electrically charged polar end and a fatty chain non-polar end, such as distearoylphosphatidylcholine, dimyristoylphosphocholine, dioleoylphosphocholine, palmitoylphosphocholine, 1,2-distearoylphosphocholine, heneicosanoylphosphocholine or palmitoylphosphocholine; (3) The lipid carrier further includes PEG lipids, which are lipid molecules modified with a polyethylene glycol hydrophilic end, such as PEG-modified dimyristoylglycerol; (4) the lipid carrier further comprises a sterol, wherein the sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol; (5) In the lipid carrier, the molar ratio of the substance Z to the sterol is 0.5-5:1, preferably 0.5-3:1, for example 1.3:1; (6) In the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 2-8:1, for example 3-6:1; (7) In the lipid carrier, the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 1-10:1, for example 5:1; (8) In the lipid carrier, the molar ratio of the substance Z to the PEG lipid is 10-100:1, preferably 10-50:1, for example 33.3:1; (9) In the lipid carrier, the molar content of the substance Z is 30 mol% to 70 mol%, for example, 50 mol%; (10) In the lipid carrier, the molar content of the phospholipid is 5 mol% to 20 mol%, for example, 10 mol%; (11) In the lipid carrier, the molar content of the sterol is 20 mol% to 60 mol%, for example, 38.5 mol%; (12) In the lipid carrier, the molar content of the PEG lipid is about 0.2 mol% to 5 mol%, for example, 1.5% mol; (13) The lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol; (14) The lipid carrier formulation is as follows: substance Z: phospholipid: sterol: PEG lipid in a ratio of 50:10:38.5:1.5; The substance Z is preferably The phospholipid is preferably DSPC; the sterol is preferably cholesterol; and the PEG lipid is preferably DMG-PEG2000.

10. Use of a compound of formula I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 or a lipid carrier according to any one of claims 8 to 9 in the preparation of a delivery vector for a nucleic acid preventive agent and / or therapeutic agent; The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid, double-stranded DNA, small interfering RNA, asymmetric double-stranded small interfering RNA, microRNA, small hairpin RNA, circular RNA, transfer RNA or messenger RNA, preferably mRNA, such as firefly luciferase mRNA, SARS-CoV-2 spike protein mRNA, herpes zoster virus mRNA, respiratory syncytial virus mRNA or influenza virus mRNA.

11. A lipid nanoparticle comprising a nucleic acid preventive and / or therapeutic agent and a lipid carrier according to any one of claims 8 to 9; The nucleic acid therapeutic and / or preventive agent is preferably one or more of single-stranded deoxyribonucleic acid, double-stranded DNA, small interfering RNA, asymmetric double-stranded small interfering RNA, microRNA, small hairpin RNA, circular RNA, transfer RNA or messenger RNA, preferably mRNA, such as firefly luciferase mRNA, SARS-CoV-2 spike protein mRNA, herpes zoster virus mRNA, respiratory syncytial virus mRNA or influenza virus mRNA.

12. The lipid nanoparticle according to claim 11, wherein 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, further preferably (3-16):1, for example 6:1; (2) The particle size of the lipid nanoparticles is 10-200 nm, preferably 40-150 nm, 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 lipid nanoparticles have a particle size (average particle size) of 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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