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, and particularly provides a compound I or pharmaceutically acceptable salt of the compound I. An LNP preparation prepared from the nitrogen-containing chain compound is relatively uniform in nanoparticle size, relatively high in encapsulation efficiency and relatively high in in-vivo expression activity. # imgabs0 #

Description

Technical Field

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

[0002] Nucleic acid drugs are an important direction of basic and applied research today. Nucleic acid drugs can be used to prevent and / or treat viral and bacterial infectious diseases, tumors, metabolic diseases, etc. Their production costs are lower and the cycle is shorter, which is conducive to the rapid development of personalized drugs. However, nucleic acids are negatively charged macromolecules that are difficult to penetrate cell membranes. At the same time, nucleic acids are not stable. By developing various nucleic acid packaging and delivery systems, the instability of nucleic acid drugs can be overcome to a certain extent and their delivery efficiency can be improved.

[0003] Lipid nanoparticles have been shown to be useful as carriers for delivering bioactive substances (such as small molecule drugs, proteins, and nucleic acids) into cells and / or intracellular compartments. Optimizing the nucleic acid drug delivery system by designing and optimizing the types and amounts of each component in the lipid nanoparticles is of great significance for improving the efficacy of nucleic acid drugs for prevention and treatment, especially lipid compounds that can be used to deliver RNA preventive and / or therapeutic agents and related methods and compositions.

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

[0005] In order to overcome the problem that the types of ionizable lipid compounds and nucleic acid preventive and / or therapeutic agent delivery carriers in the prior art are few and the effects are poor, the present invention provides a nitrogen-containing chain compound, a preparation method, a composition comprising the same and an application thereof. The composition of the present invention can be used for efficient delivery of nucleic acid drugs. The LNP preparation prepared by using the nitrogen-containing chain compound of the present invention has one or more of the following advantages: (1) high encapsulation efficiency (2) uniform particle size (3) high in vivo expression activity.

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

[0007]

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

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

[0010] Z 1 and Z 2 Independently

[0011] W 1 and W 2 Independently a chemical bond or C 1-12 Alkylene;

[0012] R 2 C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-10 Cycloalkylene-R 21 , where R 21 H or C 1-6 alkyl;

[0013] R 3 C 1-15 Alkyl, or one, two, or three CH 2 C substituted by oxygen 1-15 alkyl;

[0014] R 4 C 1-15 Alkyl, C 1-15 Alkoxy, C 1-15 Alkenyloxy, -C 1-12 Alkylene-C 3-10 Cycloalkylene-R 41 、-OC 1-12 Alkylene-C 3-10 Cycloalkylene-R 42 ; Among them, R 41 and R 42 are independently H or C 1-6 alkyl;

[0015] R 5 H, C 1-15 Alkyl, or one, two, or three CH 2 C substituted by oxygen 1-15 alkyl.

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

[0017] In one embodiment of the present invention, R 1 , R 21 , R 41 and R 42 In the 1-6 "alkyl" are each independently C 1-4 Alkyl, for example methyl, ethyl, n-propyl or n-butyl.

[0018] In one embodiment of the present invention, R 1 , R 21 , R 41 and R 42 In the 1-6 "Alkyl" is each independently a straight chain or branched C 1-6 alkyl.

[0019] In one embodiment of the present invention, R 2 and R 4 In the C 3-10 Cycloalkylene is C 3-6 Cycloalkylene, for example,

[0020] In one embodiment of the present invention, R 1 In the case of a hydroxyl group, only the terminal carbon not attached to the nitrogen atom is replaced by a hydroxyl group.

[0021] In one embodiment of the present invention, R 1 for

[0022] In one embodiment of the present invention, X and Y are independently C 3-7 Alkylene.

[0023] In one embodiment of the present invention, X and Y are independently C 3-7 Straight chain alkylene.

[0024] In a certain embodiment of the present invention, X is

[0025] In a certain embodiment of the present invention, Y is

[0026] In one embodiment of the present invention, Z 1 for *Denotes the same as W 1 connect.

[0027] In one embodiment of the present invention, Z 2 for *Denotes the same as W 2 connect.

[0028] In one embodiment of the present invention, W 1 and W 2 Independently chemically bonded or linear C 1-6 Alkylene.

[0029] In one embodiment of the present invention, W 1 for

[0030] In one embodiment of the present invention, W 2 For chemical bonds,

[0031] In one embodiment of the present invention, R 2 C 4-9 Alkyl, C 4-9 Alkenyl or -C 4-6 Alkylene-C 3-10 Cycloalkylene-R 21 .

[0032] In one embodiment of the present invention, R 2 for

[0033] In one embodiment of the present invention, R 2 for

[0034] In one embodiment of the present invention, R 3 C 5-10 Alkyl, or a CH 2 C substituted by oxygen 5-10 alkyl.

[0035] In one embodiment of the present invention, R 3 for

[0036] In one embodiment of the present invention, R 4 C 8-10 Alkyl, C 5-9 Alkoxy, C 8-10 Alkenyloxy, -C 1-3 Alkylene-C 3-10 Cycloalkylene-R 41 、-OC 4-6 Alkylene-C 3-10 Cycloalkylene-R 42 .

[0037] In one embodiment of the present invention, R 4 for

[0038] In one embodiment of the present invention, R 4 for

[0039] In one embodiment of the present invention, R 5 H, C 8-10 Alkyl, or a CH 2 C substituted by oxygen 8-10 alkyl.

[0040] In one embodiment of the present invention, R 5 For H.

[0041] In one embodiment of the present invention,

[0043] In one embodiment of the present invention,

[0045] In one embodiment of the present invention,

[0047] In one embodiment of the present invention, the compound I has the following structure:

[0048]

[0049] 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 any of the previous schemes. In one embodiment of the present invention, the compound I is any of the following structures:

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056] The present invention also provides a use of the compound I or a pharmaceutically acceptable salt thereof in the preparation of a nucleic acid preventive agent and / or therapeutic agent delivery vector;

[0057] The nucleic acid preventive and / or therapeutic 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.

[0058] The present invention also provides a composition comprising a substance Z, wherein the substance Z is the compound I as described above or a pharmaceutically acceptable salt thereof.

[0059] The present invention also provides a lipid carrier, which includes a substance Z, wherein the substance Z is the compound I as described above or a pharmaceutically acceptable salt thereof.

[0060] In one embodiment of the present invention, the lipid carrier further comprises a diluent, which can be a phosphate buffer, a sodium acetate buffer or a Tris-acetate buffer.

[0061] In one embodiment of the present invention, the lipid carrier further comprises phospholipids.

[0062] In a certain 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 an aliphatic chain, such as distearoylphosphatidylcholine (DSPC), dimyristoylphosphocholine (DMPC), dioleoylphosphocholine (DOPC), palmitoylphosphocholine (DPPC), heneicosanoylphosphocholine (DUPC) or palmitoylphosphocholine (POPC), etc., preferably distearoylphosphatidylcholine.

[0063] In a certain embodiment of the present invention, the lipid carrier further comprises PEG lipid (lipid modified with polyethylene glycol).

[0064] In a certain embodiment of the present invention, the PEG lipid may be a lipid molecule modified with a polyethylene glycol hydrophilic end. The PEG lipid is preferably selected from one or more of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol and PEG-modified dialkylglycerol, such as PEG-modified dimyristoylglycerol (DMG-PEG2000).

[0065] In one embodiment of the present invention, the lipid carrier further comprises sterol.

[0066] In a certain embodiment of the present invention, the sterol can be a conventional sterol in the art, and the sterol includes animal, plant or fungal sterols. The sterol is selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol.

[0067] In a certain embodiment of the present invention, in the lipid carrier, the molar ratio of the substance Z to sterol is 0.5-5:1, preferably 0.5-3:1, such as 1.3:1.

[0068] In a certain 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.

[0069] 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.

[0070] In a certain 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.

[0071] In the present invention, the 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 shall not exceed 100 mol%.

[0072] 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%.

[0073] In one embodiment of the present invention, in the lipid carrier, the molar content of the phospholipid is 5 mol% to 20 mol%, for example, 10 mol%.

[0074] 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%.

[0075] In one embodiment of the present invention, in the lipid carrier, the molar content of the PEG lipid is about 0.2 mol% to 5 mol%, for example, 1.5% mol.

[0076] In a certain embodiment of the present invention, the lipid carrier is composed of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.

[0077] The present invention also provides a use of the lipid carrier in preparing a nucleic acid preventive agent and / or therapeutic agent delivery vector;

[0078] The nucleic acid preventive and / or therapeutic 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.

[0079] The present invention also provides a lipid nanoparticle, which comprises a nucleic acid preventive agent and / or a therapeutic agent and the aforementioned lipid carrier;

[0080] The nucleic acid preventive and / or therapeutic 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.

[0081] In a certain embodiment of the present invention, the nitrogen-to-phosphorus ratio in the lipid nanoparticles may be (2-30):1, and the nitrogen-to-phosphorus ratio refers to the ratio of the molar number of ionizable nitrogen atoms in one or more ionizable lipid compounds to the molar number of phosphate groups in RNA, and in the present application, it refers to the ratio of the molar number of ionizable nitrogen atoms in the ionizable lipid nanoparticles to the molar number of phosphate groups in mRNA in the pharmaceutical composition. Preferably, the nitrogen-to-phosphorus ratio is (2-20):1, preferably (3-15):1, for example 6:1.

[0082] In one embodiment of the present invention, the particle size (average particle size) of the lipid nanoparticles is 10-200 nm, preferably 40-150 nm, and more preferably 50-130 nm, such as 67.67 nm, 53.47 nm or 61.92 nm.

[0083] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.001-0.3, for example, 0.086, 0.069, or 0.035.

[0084] In a certain embodiment of the present invention, the encapsulation efficiency of the lipid nanoparticles is 90%-100%, preferably 94%-100%, such as 94.94%, 96.14%, 96.62%.

[0085] In a certain embodiment of the present invention, the particle size (average particle size) of the lipid nanoparticles is 10-200nm, preferably 40-150nm, and more preferably 50-130nm, for example, 53.4nm, 61.9nm, 66.1nm, 81.0nm, 84.0nm, 78.8nm, 57.0nm, 60.6nm, 61.4nm, 94.6nm, 81.7nm, 58.1nm, 67.8nm, 72.1nm, 57.8nm, 55.4nm, 60.7nm or 64.4nm.

[0086] In one embodiment of the present invention, the polydispersity index of the lipid nanoparticles is 0.001-0.3, for example, 0.069, 0.035, 0.030, 0.088, 0.080, 0.075, 0.077, 0.063, 0.055, 0.034, 0.169, 0.112, 0.038, 0.193, 0.178, 0.084, 0.050 or 0.067.

[0087] In one embodiment of the present invention, the encapsulation efficiency of the lipid nanoparticles is 89%-100%, for example 89-98%; for example 96.1%, 96.6%, 96.8%, 93.0%, 94.7%, 94.0%, 95.4%, 96.3%, 97.4%, 91.3%, 89.7%, 97.6%, 96.9%, 96.2% or 96.4%.

[0088] In one embodiment of the present invention, in the lipid nanoparticles, the lipid carrier encapsulates the nucleic acid preventive agent and / or therapeutic agent.

[0089] On the basis of being in accordance with the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0090] The reagents and raw materials used in the present invention are commercially available.

[0091] Unless otherwise specified, the terms used in the present invention have the following meanings:

[0092] The positive and progressive effects of the present invention are 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.

[0093] The term "alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms (e.g., C1 to C6). Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0094] The term "alkylene" refers to a group formed by losing a hydrogen atom from the aforementioned "alkyl".

[0095] The term "cycloalkyl" refers to a saturated monocyclic ring group consisting only of carbon atoms with a specified number of carbon atoms (e.g., C3-C6). Cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0096] The term "cycloalkylene" refers to a group formed by losing a hydrogen atom from the aforementioned "cycloalkyl".

[0097] The term "alkenyloxy" refers to a group of the structure -O-alkenyl, for example, C 1-15 Alkenyloxy refers to -OC 1-15 Alkenyl, wherein "alkenyl" refers to a straight or branched chain hydrocarbon group containing one or more carbon-carbon double bonds (eg, 1, 2 or 3 carbon-carbon double bonds).

[0098] The term "treat" refers to any of the following: (1) alleviating one or more biological manifestations of a disease; (2) interfering with one or more points in the biological cascade that initiates a disease; or (3) slowing the progression of one or more biological manifestations of a disease.

[0099] The term "prevention" refers to reducing the risk of developing a disease.

[0100] The positive and progressive effects of the present invention are 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

[0101] Figure 1 This is a graph showing the change in chemiluminescence intensity over time in the liver area of ​​mice after intravenous administration of compound 1 and compound 2;

[0102] Figure 2 This is a graph showing the change in chemiluminescence intensity over time in the liver region of mice after intravenous administration of Compound 4, Compound 6, Compound 7, Compound 10-Compound 13, Compound 15, Compound 20, and Compound 21;

[0103] Figure 3 This is a graph showing the change in chemiluminescence intensity over time in the liver region of mice after intravenous administration of Compound 5, Compound 8, Compound 9, Compound 14, Compound 22, and Compound 23;

[0104] Figure 4 This is a graph showing the change in chemiluminescence intensity over time in the liver area of ​​mice after intravenous administration of compound 16 and compound 24. DETAILED DESCRIPTION

[0105] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0106] Preparation Example 1 Preparation of Compound 1

[0107] Step 1: Synthesis of 1-(benzyloxy)undecane-2-ol

[0108] Reaction:

[0109]

[0110] Material ratio:

[0111] Material Name Molecular weight Feed ratio Feeding amount mmol 2-Benzyloxyacetaldehyde 150 1.0 eq 4.00g 26.6mmol Nonylmagnesium bromide 230 1.1eq 6.78g 29.3mmol Tetrahydrofuran - - 20mL -

[0112] Operation process:

[0113] Two batches were reacted in parallel. 2-Benzyloxyacetaldehyde (4.00 g, 26.6 mmol) was dissolved in tetrahydrofuran (20 mL), and a tetrahydrofuran solution of nonylmagnesium bromide (6.78 g, 29.3 mmol) was slowly added dropwise, and the mixture was reacted at 20°C for 12 hours under nitrogen protection. The reaction was completed by TLC monitoring, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid 1-(benzyloxy)undecane-2-ol (6.00 g, 66.0% yield).

[0114] Step 2: Synthesis of ({[2-(octyloxy)undecyl]oxy}methyl)benzene

[0115] Reaction:

[0116]

[0117] Material ratio:

[0118] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)undecane-2-ol 278 1.0 eq 4.50g 16.2mmol Octane bromide 192 1.5eq 4.68g 24.2mmol Sodium hydride (60%) 24 2.5eq 1.62g 40.4mmol N,N-Dimethylformamide - - 90mL -

[0119] Operation process:

[0120] Two batches were reacted in parallel. 1-(Benzyloxy)undecane-2-ol (4.50 g, 16.2 mmol) and sodium hydride (1.62 g, 40.4 mmol, 60%) were dissolved in N, N-dimethylformamide (45 mL), stirred at 20°C for 0.5 hours under nitrogen protection, and a solution of n-octane bromide (4.68 g, 24.2 mmol) in N, N-dimethylformamide (45 mL) was slowly added dropwise at 0°C. Finally, the reaction was carried out at 50°C for 6 hours under nitrogen protection, and the reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[2-(octyloxy)undecyl]oxy}methyl)benzene (6.00 g, 77.8% yield).

[0121] Step 3: Synthesis of 2-(octyloxy)undecane-1-ol

[0122] Reaction:

[0123]

[0124] Material ratio:

[0125]

[0126] Operation process:

[0127] Two batches were reacted in parallel. ({[2-(octyloxy)undecyl]oxy}methyl)benzene (5.50 g, 14.1 mmol) was dissolved in methanol (50 mL) and tetrahydrofuran (5 mL), palladium / carbon (2.80 g, 2.63 mmol) was added under nitrogen atmosphere, and finally reacted at 35°C for 12 hours under hydrogen (40 Psi) atmosphere. The reaction was completed after TLC monitoring, and the reaction solution was filtered and concentrated. The crude product was subjected to column chromatography to obtain colorless liquid 2-(octyloxy)undecane-1-ol (4.21 g, 91.2% yield).

[0128] Step 4: Synthesis of 2-(octyloxy)undecyl 5-bromopentyl ester

[0129] Reaction:

[0130]

[0131] Material ratio:

[0132]

[0133]

[0134] Operation process:

[0135] 2-(Octyloxy)undecane-1-ol (4.00 g, 13.3 mmol) was dissolved in dichloromethane (40 mL), and 5-bromovaleric acid (3.13 g, 17.3 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.83 g, 20.0 mmol), and 4-dimethylaminopyridine (163 mg, 1.33 mmol) were added in sequence. Finally, the mixture was reacted at 20° C. for 12 hours under nitrogen protection. The reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain colorless liquid 2-(Octyloxy)undecyl 5-bromopentyl ester (5.20 g, 84.3% yield).

[0136] Step 5: Synthesis of heptadecan-9-yl 8-[(2-hydroxyethyl)(5-{[2-(octyloxy)undecyl]oxy}-5-oxypentylene)amino]octyl ester

[0137] Reaction:

[0138]

[0139] Material ratio:

[0140]

[0141] Operation process:

[0142] The reaction was carried out in parallel in 2 batches. 2-(Octyloxy)undecyl 5-bromopentyl ester (1.00 g, 2.26 mmol) was dissolved in acetonitrile (8 mL), and heptadecan-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (1.10 g, 2.38 mmol), potassium carbonate (1.10 g, 7.92 mmol), potassium iodide (451 mg, 2.72 mmol), tetrahydrofuran (2 mL) were added in sequence, and finally reacted at 80° C. for 12 hours under nitrogen protection, and the reaction was monitored by TLC, filtered, and concentrated. The light yellow liquid heptadecan-9-yl 8-[(2-hydroxyethyl)(5-{[2-(octyloxy)undecyl]oxy}-5-oxyylidenepentyl)amino]octyl ester (800 mg, 26.5% yield) was obtained by purification by column chromatography.

[0143] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.87 (quin, J = 6.2Hz, 1H), 4.16-4.10 (m, 1H), 4.07-4.00 (m, 1H), 3.62-3.51 (m, 3H), 3.47-3.38 (m, 2H), 2.64 (br t,J=5.2Hz,2H),2.57-2.49(m,4H),2.36 (t,J=7.2Hz,2H),2.28(t,J=7.4Hz,2H),1.68-1.59(m,4H),1.57-1.45 (m,12H),1.26(br s,54H),0.98 -0.79(m,12H)ppm.

[0144] LCMS:RT=2.307,m / z 824.6[M+H] + .

[0145] Preparation Example 2 Preparation of Compound 2

[0146] Step 1: Synthesis of 1-(Benzyloxy)dodecan-2-ol

[0147] Reaction:

[0148]

[0149] Material ratio:

[0150] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Benzyloxy)acetaldehyde 150 1.0eq 5.0g 33.29mmol Decylmagnesium bromide 244 1.2eq 9.81g 39.95mmol Tetrahydrofuran - - 50mL -

[0151] Operation process:

[0152] 2-(Benzyloxy)acetaldehyde (5.0 g, 33.29 mmol) was dissolved in tetrahydrofuran (50 mL), and bromo(decyl)magnesium (9.81 g, 0.5 M, 79.91 mL, 19.06 mmol) was added at 0°C, and the mixture was reacted at 25°C for 12 hours under nitrogen protection. After the reaction was completed by TLC monitoring, 50 mL of ammonium chloride aqueous solution was added, and the mixture was extracted with ethyl acetate twice, each time with 200 mL. The organic phase was concentrated and purified by column chromatography to obtain colorless liquid 1-(benzyloxy)dodecan-2-ol (7.5 g, 77.0% yield).

[0153] Step 2: Synthesis of ({[2-(octyloxy)dodecyl]oxy}methyl)benzene

[0154] Reaction:

[0155]

[0156] Material ratio:

[0157] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-2-ol 292 1.0 eq 7.00g 23.9mmol 1-Bromooctane 192 1.2eq 5.55g 28.7mmol Sodium Hydrogen 24 2.0eq 1.91g 47.9mmol 1-Methyl-2-pyrrolidone - - 70mL -

[0158] Operation process:

[0159] 1-(Benzyloxy) dodecan-2-ol (7.00 g, 23.94 mmol) was dissolved in 1-methyl-2-pyrrolidone (70 mL), sodium hydrogen (1.91 g, 47.87 mmol) was added under nitrogen protection at 0°C, and the reaction was carried out at 0°C for 1 hour. 1-Bromooctane (5.55 g, 28.72 mmol) was added to the reaction solution, and the reaction was carried out at 20°C under nitrogen protection for 12 hours. TLC monitoring showed that new spots were generated in the reaction, 15 mL of water was added, and the mixture was extracted with ethyl acetate twice, each time with 20 mL, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[2-(octyloxy) dodecanyl] oxy} methyl) benzene (6.7 g, 69.1% yield).

[0160] Step 3: Synthesis of 2-(octyloxy)dodecan-1-ol

[0161] Reaction:

[0162]

[0163] Material ratio:

[0164]

[0165] Operation process:

[0166] Dissolve ({[2-(octyloxy) dodecanyl] oxy} methyl) benzene (6.70 g, 16.56 mmol) in methanol (70 mL), add palladium carbon (3.52 g, 0.2 mmol), react at 35°C, 40 Psi under hydrogen protection for 12 hours, monitor the completion of the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain colorless liquid 2-(octyloxy) dodecan-1-ol (3.90 g, 74.8% yield).

[0167] Step 4: Synthesis of 2-(octyloxy)dodecyl 6-bromohexyl ester

[0168] Reaction:

[0169]

[0170] Material ratio:

[0171]

[0172] Operation process:

[0173] 2-(Octyloxy) dodecan-1-ol (3.9 g, 12.4 mmol), 6-bromohexanoic acid (2.66 g, 13.6 mmol), dissolved in dichloromethane (80 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.57 g, 18.6 mmol), 4-dimethylaminopyridine (151 mg, 1.24 mmol) were added in sequence, and finally reacted at 25°C for 12 hours under nitrogen protection. TLC monitored the reaction to see new spots, added 10 mL of water, extracted twice with dichloromethane, 10 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain bright yellow liquid 2-(octyloxy) dodecan-6-bromohexyl ester (4.80 g, 78.7% yield).

[0174] Step 5: Synthesis of 2-(octyloxy)dodecyl 6-[(2-hydroxyethyl)(6-{[2-(octyloxy)dodecyl]oxy}-6-oxyylidenehexyl)amino]hexyl ester

[0175] Reaction:

[0176]

[0177] Material ratio:

[0178] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Octyloxy)dodecyl 6-bromohexyl ester 490 1.00eq 800mg 1.63mmol 2-Aminoethane-1-ol 61.05 0.45eq 44.73mg 0.73mmol Potassium carbonate 138 3.0 eq 674mg 4.88mmol Potassium iodide 166 1.5 eq 405mg 2.44mmol Acetonitrile - - 1.5mL - Tetrahydrofuran - - 1.5mL -

[0179] Operation process:

[0180] 2-(Octyloxy) dodecyl 6-bromohexyl ester (800 mg, 1.63 mmol) was dissolved in acetonitrile (1.5 mL), and 2-aminoethane-1-ol (44.73 mg, 0.73 mmol), potassium carbonate (674 mg, 4.88 mmol), potassium iodide (405 mg, 2.44 mmol), and tetrahydrofuran (1.5 mL) were added in sequence. Finally, the mixture was reacted at 75° C. for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Column chromatography was used to purify the mixture to obtain a bright yellow liquid 2-(octyloxy) dodecyl 6-[(2-hydroxyethyl)(6-{[2-(octyloxy) dodecyl]oxy}-6-oxyylidenehexyl)amino]hexyl ester (280 mg, 18.1% yield).

[0181] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.17-4.10 (m, 2H), 4.08-4.00 (m, 2H), 3.61-3.52 (m, 4H), 3.47-3.39 (m, 4H), 2.63 (br s, 2H), 2.51 (br t,J=6.6Hz,4H),2.34(t,J=7.4Hz,4H),1.65(td,J 1 =7.6,J 2=15.2Hz,5H),1.59-1.42(m,14H),1.39-1.23(m,54H),0.93-0.85(m,11H)ppm.

[0182] LCMS:RT=2.231,m / z 883.4[M+H] + .

[0183] Preparation Example 3 Preparation of Compound 4

[0184] Step 1: Synthesis of nonyl 6-[(2-hydroxyethyl)amino]hexyl ester

[0185] Reaction:

[0186]

[0187] Material ratio:

[0188]

[0189]

[0190] Operation process:

[0191] Dissolve nonyl 6-bromohexyl ester (1.00 g, 3.11 mmol) in acetonitrile (10.0 mL), add ethanolamine (1.90 g, 31.1 mmol) in turn, and react at 30 ° C for 12 hours under nitrogen protection. After TLC monitoring, the reaction is complete, filtered, and concentrated. Purify by column chromatography to obtain colorless liquid nonyl 6-[(2-hydroxyethyl)amino]hexyl ester (650 mg, 69.2% yield).

[0192] Step 2: Synthesis of 2-(octyloxy)dodecyl 5-bromopentyl ester

[0193] Reaction:

[0194]

[0195] Material ratio:

[0196]

[0197] Operation process:

[0198] 2-(Octyloxy) dodecan-1-ol (450 mg, 1.43 mmol), 5-bromovaleric acid (362 mg, 2.00 mmol), dissolved in dichloromethane (4.50 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (493 mg, 2.58 mmol), 4-dimethylaminopyridine (34.9 mg, 286 μmol) were added in sequence, and finally reacted at 25 ° C for 12 hours under nitrogen protection. TLC monitoring showed that new spots were generated in the reaction, 10 mL of water was added, and dichloromethane was extracted twice, 10 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain bright yellow liquid 2-(Octyloxy) dodecan-5-bromopentyl ester (420 mg, 61.4% yield).

[0199] Step 3: Synthesis of nonyl 6-[(2-hydroxyethyl)(5-{[2-(octyloxy)dodecyl]oxy}-5-oxyylidenepentyl)amino]hexyl ester

[0200] Reaction:

[0201]

[0202] Material ratio:

[0203]

[0204]

[0205] Operation process:

[0206] 2-(Octyloxy) dodecyl 5-bromopentyl ester (340 mg, 711 μmol) was dissolved in acetonitrile (4.00 mL), and nonyl 6-[(2-hydroxyethyl) amino] hexyl ester (214 mg, 711 μmol), potassium carbonate (344 mg, 2.49 mmol), potassium iodide (141 mg, 854 μmol), tetrahydrofuran (2.00 mL) were added in sequence, and finally reacted at 75°C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a colorless liquid nonyl 6-[(2-hydroxyethyl)(5-{[2-(octyloxy) dodecyl] oxy}-5-oxyylidenepentyl) amino] hexyl ester (356 mg, 67.7% yield).

[0207] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.18-4.10 (m, 1H), 4.09-3.98 (m, 3H), 3.63-3.51 (m, 3H), 3.49-3.35 (m, 2H), 2.65 (br t,J=5.2Hz,2H),2.59-2.48(m,4H),2.41-2.25(m,4H),1.66-1.59(m,6H),1.56-1.43(m,8H),1.37-1.17(m,40H),0.89(t,J=6.8Hz,9H)ppm.

[0208] LCMS:RT=1.985,m / z 698.6[M+H] + .

[0209] Preparation Example 4 Preparation of Compound 6

[0210] Step 1: Synthesis of ethyl 2-(4-pentylcyclohexylidene) acetate

[0211] Reaction:

[0212]

[0213] Material ratio:

[0214] Material Name Molecular weight Feed ratio Feeding amount mmol 4-Pentylcyclohexane-1-one 168 1.0 eq 5.00g 29.7 Triethyl phosphoacetate 224 1.1eq 7.33g 32.7 Sodium hydride 24 1.1eq 1.31g 32.7 Tetrahydrofuran - - 50mL -

[0215] Operation process:

[0216] Dissolve 4-pentylcyclohexane-1-one (5.00 g, 29.7 mmol) and sodium hydride (1.31 g, 32.7 mmol, 60%) in tetrahydrofuran (25 mL), stir at 20°C for 0.5 hours under nitrogen protection, slowly dropwise add triethyl phosphoacetate (7.33 g, 32.7 mmol) in N, N-dimethylformamide (25 mL) at 0°C, and finally react at 20°C for 1.5 hours under nitrogen protection. The reaction is completed by TLC monitoring, and the reaction solution is washed with water, extracted, and concentrated. The crude product is subjected to column chromatography to obtain colorless liquid ethyl 2-(4-pentylcyclohexylidene) acetate (6.50 g, 61.7% yield).

[0217] Step 2: Synthesis of ethyl 2-(4-pentylcyclohexyl) acetate

[0218] Reaction:

[0219]

[0220] Material ratio:

[0221]

[0222]

[0223] Operation process:

[0224] The reaction was carried out in parallel in 1.45 batches. Ethyl 2-(4-pentylcyclohexylene) acetate (4.50 g, 18.9 mmol) was dissolved in methanol (40 mL), palladium / carbon (2.61 g, 2.45 mmol) was added under a nitrogen atmosphere, and finally reacted at 35° C. for 12 hours under a hydrogen (40 Psi) atmosphere. The reaction was completed by TLC monitoring, and the reaction liquid was filtered and concentrated. The crude product was subjected to column chromatography to obtain colorless liquid ethyl 2-(4-pentylcyclohexyl) acetate (6.00 g, 99.1% yield).

[0225] Step 3: Synthesis of 2-(4-pentylcyclohexyl)ethane-1-ol

[0226] Reaction:

[0227]

[0228] Material ratio:

[0229] Material Name Molecular weight Feed ratio Feeding amount mmol Ethyl 2-(4-pentylcyclohexyl) acetate 240 1.0 eq 5.00g 20.8 Lithium Aluminum Tetrahydride 38 2.00eq 1.58g 41.6 Tetrahydrofuran - - 50mL -

[0230] Operation process:

[0231] Dissolve ethyl 2-(4-pentylcyclohexyl) acetate (5.00 g, 20.8 mmol) in tetrahydrofuran (50 mL), slowly add a tetrahydrofuran solution of lithium aluminum tetrahydride (1.58 g, 41.6 mmol, 2.5 M) at 0°C, and react at 25°C for 12 hours under nitrogen protection. The reaction is completed by TLC monitoring, and the reaction solution is concentrated. The crude product is purified by column chromatography to obtain a colorless liquid 2-(4-pentylcyclohexyl)ethane-1-ol (4.00 g, 96.9% yield).

[0232] Step 4: Synthesis of 2-(4-pentylcyclohexyl)ethyl 8-bromooctyl ester

[0233] Reaction:

[0234]

[0235] Material ratio:

[0236]

[0237] Operation process:

[0238] 2-(4-pentylcyclohexyl)ethane-1-ol (1.00 g, 34.2 mmol) was dissolved in dichloromethane (10 mL), and 8-bromooctanoic acid (1.46 g, 44.5 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.45 g, 51.3 mmol), and 4-dimethylaminopyridine (61.6 mg, 3.42 mmol) were added in sequence. Finally, the mixture was reacted at 25° C. for 12 hours under nitrogen protection. The reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid 2-(4-pentylcyclohexyl)ethyl 8-bromooctyl ester (1.60 g, 78.6% yield).

[0239] Step 5: Synthesis of 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)amino]octyl ester

[0240] Reaction:

[0241]

[0242] Material ratio:

[0243] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(4-Pentylcyclohexyl)ethyl 8-bromooctyl ester 4.3 1.0 eq 1.60g 3.97 Ethanolamine 61 10eq 2.40g 39.7 Acetonitrile - - 20mL -

[0244] Operation process:

[0245] 2-(4-pentylcyclohexyl)ethyl 8-bromooctyl ester (10.g, 3.97mmol) and ethanolamine (2.40g, 39.7mmol) were dissolved in acetonitrile (20mL) and reacted at 25°C for 12 hours under nitrogen atmosphere. The reaction was completed after monitoring by TLC. The reaction solution was washed with water, extracted, and concentrated. The crude product was purified by column chromatography to obtain a colorless liquid 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)amino]octyl ester (1.10g, 72.3% yield).

[0246] Step 6: Synthesis of 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)(8-{[2-(octyloxy)undecyl]oxy}-8-oxyylideneoctyl)amino]octyl ester

[0247] Reaction:

[0248]

[0249] Material ratio:

[0250]

[0251] Operation process:

[0252] 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)amino]octyl ester (1.30 g, 2.58 mmol) was dissolved in acetonitrile (12 mL), and 2-(octyloxy)undecyl 8-bromooctyl ester (900 mg, 2.35 mmol), potassium carbonate (1.13 g, 8.21 mmol), potassium iodide (467 mg, 2.28 mmol), and tetrahydrofuran (3 mL) were added in sequence. Finally, the mixture was reacted at 75° C. for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. The mixture was purified by column chromatography to obtain a light yellow liquid 2-(4-pentylcyclohexyl)ethyl 8-[(2-hydroxyethyl)(8-{[2-(octyloxy)undecyl]oxy}-8-oxyylideneoctyl)amino]octyl ester (750 mg, 39.6% yield).

[0253] 1 H NMR(400MHz,CHLOROFORM-d)δ=4.15-4.07(m,3H),4.06-4.01(m,1H),3.60-3.51 (m,3H),3.47-3.39(m,2H),2.61(t,J=5.2Hz,2H),2.52-2.45(m,4H),2.31(td,J 1 =7.4,J 2 =14.9Hz,4H),1.74(br d,J=10.4Hz,3H),1.66-1.58(m,5H),1.56-1.42(m,11H),1.29(br d,J=14.0Hz,44H),1.18-1.13(m,2H),1.00-0.79(m,12H)ppm.

[0254] LCMS:RT=2.250,m / z 808.7[M+H] + .

[0255] Preparation Example 5 Preparation of Compound 7

[0256] Step 1: Synthesis of 2-(octyloxy)dodecyl 4-bromobutyl ester

[0257] Reaction:

[0258]

[0259] Material ratio:

[0260]

[0261] Operation process:

[0262] 2-(Octyloxy) dodecan-1-ol (1.00 g, 3.18 mmol), 4-bromobutyric acid (584 mg, 3.50 mmol), dissolved in dichloromethane (10 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (914 mg, 4.77 mmol), 4-dimethylaminopyridine (38.8 mg, 0.318 mmol) were added in sequence, and finally reacted at 25°C for 12 hours under nitrogen protection. TLC monitored the reaction and new spots were generated. 10 mL of water was added, and dichloromethane was extracted twice, 10 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain bright yellow liquid 2-(Octyloxy) dodecan-4-bromobutyl ester (1.00 g, 67.8% yield).

[0263] Step 2: Synthesis of 2-(octyloxy)dodecyl 6-[(2-hydroxyethyl)amino]hexyl ester

[0264] Reaction:

[0265]

[0266] Material ratio:

[0267] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Octyloxy)dodecyl 4-bromobutyl ester 490 1.0 eq 1.80g 3.66 2-Aminoethane-1-ol 61.05 10.0eq 2.24g 36.6 Acetonitrile - - 20mL -

[0268] Operation process:

[0269] 2-(Octyloxy) dodecyl 4-bromobutyl ester (1.80 g, 3.66 mmol) was dissolved in acetonitrile (20 mL), and 2-aminoethane-1-ol (2.24 g, 36.6 mmol) was added in sequence, and the mixture was reacted at 25° C. for 24 hours under nitrogen protection. The reaction was completed after TLC monitoring, and the mixture was filtered and concentrated. Column chromatography was used for purification to obtain a bright yellow liquid 2-(Octyloxy) dodecyl 6-[(2-hydroxyethyl)amino]hexyl ester (180 mg, 10.4% yield).

[0270] Step 3: Synthesis of 2-(octyloxy)dodecyl 6-[(2-hydroxyethyl)(4-{[2-(octyloxy)dodecyl]oxy}-4-oxyylidenebutyl)amino]hexyl ester

[0271] Reaction:

[0272]

[0273] Material ratio:

[0274]

[0275] Operation process:

[0276] 2-(Octyloxy) dodecyl 4-bromobutyl ester (212 mg, 0.457 mmol) was dissolved in acetonitrile (2.0 mL), and 2-(Octyloxy) dodecyl 6-[(2-hydroxyethyl) amino] hexyl ester (180 mg, 0.381 mmol), potassium carbonate (184 mg, 1.34 mmol), potassium iodide (76.0 mg, 0.457 mmol), tetrahydrofuran (2.0 mL) were added in sequence, and finally reacted at 70° C. for 12 hours under nitrogen protection, and the reaction was completed after TLC monitoring, filtration, and concentration. Column chromatography purification gave a bright yellow liquid 2-(Octyloxy) dodecyl 6-[(2-hydroxyethyl)(4-{[2-(Octyloxy) dodecyl] oxy}-4-oxyylidenebutyl) amino] hexyl ester (160 mg, 44.7% yield).

[0277] 1 H NMR (400MHz, CHLOROFORM-d) δ=4.18-4.09(m,2H),4.07-4.01(m,2H),3.64-3.51(m,4H),3.43(td,J=6.8,8.8Hz,4H),2.67(br d,J=4.4Hz,2H),2.61-2.50(m,4H),2.36(td,J=7.2,12.0Hz,4H),1.89-1.79(m,2 H),1.70-1.63(m,4H),1.54-1.46(m,9H),1.37-1.21(m,54H),0.93-0.84(m,12H).

[0278] LCMS:RT=2.335,m / z 854.7[M+H] + .

[0279] Preparation Example 6 Preparation of Compound 10

[0280] Step 1: Synthesis of ({[2-(hexyloxy)undecyl]oxy}methyl)benzene

[0281] Reaction:

[0282]

[0283] Material ratio:

[0284]

[0285]

[0286] Operation process:

[0287] Dissolve 1-(Benzyloxy) dodecan-2-ol (5.00 g, 17.1 mmol) and sodium hydride (2.26 g, 56.4 mmol, 60%) in 1-methyl-2-pyrrolidone (60 mL), stir at 25°C for 1 hour under nitrogen protection, slowly add a solution of bromo-n-hexane (4.23 g, 25.6 mmol) in 1-methyl-2-pyrrolidone (20 mL) at 25°C. React at 25°C for 12 hours under nitrogen protection, and finally react at 50°C for 6 hours under nitrogen protection. The reaction is completed by TLC monitoring, and the reaction solution is quenched, washed with water, extracted, and concentrated. The crude product is subjected to column chromatography to obtain a colorless liquid ({[2-(hexyloxy) undecyl] oxy} methyl) benzene (4.20 g, 65.2% yield).

[0288] Step 2: Synthesis of 2-(hexyloxy)dodecan-1-ol

[0289] Reaction:

[0290]

[0291] Material ratio:

[0292]

[0293] Operation process:

[0294] Dissolve ({[2-(Hexyloxy)undecyl]oxy}methyl)benzene (4.20 g, 11.5 mmol) in methanol (40 mL), add palladium / carbon (1.190 g, 1.15 mmol) under nitrogen atmosphere, and finally react at 35°C for 12 hours under hydrogen (40 Psi) atmosphere. The reaction is completed by TLC monitoring, and the reaction liquid is filtered and concentrated. The crude product is purified by column chromatography to obtain colorless liquid 2-(hexyloxy)dodecan-1-ol (3.05 g, 95.5% yield).

[0295] Step 3: Synthesis of 2-(hexyloxy)dodecyl 6-bromohexyl ester

[0296] Reaction:

[0297]

[0298] Material ratio:

[0299]

[0300] Operation process:

[0301] 2-(Hexyloxy) dodecan-1-ol (3.05 g, 10.6 mmol) was dissolved in dichloromethane (30 mL), and 6-bromohexanoic acid (2.70 g, 13.8 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.06 g, 16.0 mmol), and 4-dimethylaminopyridine (130 mg, 1.06 mmol) were added in sequence. Finally, the mixture was reacted at 25° C. for 12 hours under nitrogen protection. The reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid 2-(hexyloxy) dodecyl 6-bromohexyl ester (4.00 g, 81.1% yield).

[0302] Step 4: Synthesis of 2-(hexyloxy)dodecyl 6-[(6-{[2-(hexyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(3-hydroxypropyl)amino]hexyl ester

[0303] Reaction:

[0304]

[0305] Material ratio:

[0306] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Hexyloxy)dodecyl 6-bromohexyl ester 463 2.0 eq 1.20g 2.59 3-Aminopropanol 75 0.98eq 95.3mg 1.27 Potassium carbonate 138 7.0eq 1.25g 9.06 Potassium iodide 166 2.3 eq 494mg 298 Acetonitrile - - 12mL - Tetrahydrofuran - - 3mL -

[0307] Operation process:

[0308] 2-(Hexyloxy) dodecyl 6-bromohexyl ester (1.20 g, 2.59 mmol) was dissolved in acetonitrile (12 mL), and 3-aminopropanol (95.3 mg, 1.27 mmol), potassium carbonate (1.25 g, 9.06 mmol), potassium iodide (494 mg, 298 mmol), tetrahydrofuran (3 mL) were added in sequence, and finally reacted at 70 ° C for 12 hours under nitrogen protection, and the reaction was completed by TLC monitoring, filtered, and concentrated. The light yellow liquid 2-(hexyloxy) dodecyl 6-[(6-{[2-(hexyloxy) dodecyl] oxo}-6-oxyylidene hexyl) (3-hydroxypropyl) amino] hexyl ester (540 mg, 46.9% yield) was obtained by column chromatography purification.

[0309] 1 H NMR(400MHz,CHLOROFORM-d)δ=4.15-4.00(m,4H),3.79(t,J=5.0Hz,2H),3.55(td,J 1 =6.6,J 2= 9.0 Hz, 2H), 3.46 - 3.39 (m, 4H), 2.69 (br t, J = 5.4 Hz, 2H), 2.52 - 2.43 (m, 4H), 2.34 (t, J = 7.4 Hz, 4H), 1.74 - 1.63 (m, 6H), 1.59 - 1.42 (m, 14H), 1.36 - 1.23 (m, 46H), 0.88 (dt, J 1 = 2.6, J 2 = 6.8 Hz, 12H) ppm.

[0310] LCMS: RT = 2.243, m / z 840.7 [M + H] + .

[0311] Preparation Example 7 Preparation of Compound 11

[0312] Step 1: Synthesis of 2-(hexyloxy)dodecyl 6-[(6-{[2-(hexyloxy)dodecyl]oxo}-6-oxohexyl)(4-hydroxybutyl)amino]hexanoate

[0313] Reaction formula:

[0314]

[0315] Material ratio:

[0316] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Hexyloxy)dodecyl 6-bromohexyl ester 463 2.0 eq 1.00g 2.16 4-Aminobutan-1-ol 8 0.98eq 94.2mg 1.06 Potassium carbonate 138 7.0eq 1.04g 7.55 Potassium iodide 166 2.3 eq 412mg 2.48 Acetonitrile - - 13mL - Tetrahydrofuran - - 2mL -

[0317] Operation process:

[0318] 2-(hexyloxy)dodecyl 6-bromohexanoate (1.00 g, 2.16 mmol) was dissolved in acetonitrile (13 mL), and 4-aminobutan-1-ol (94.2 mg, 1.06 mmol), potassium carbonate (1.04 g, 7.55 mmol), potassium iodide (412 mg, 2.48 mmol), and tetrahydrofuran (2 mL) were added in sequence. Finally, the reaction was carried out at 70 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC until completion, filtered, and concentrated. 2-(hexyloxy)dodecyl 6-[(6-{[2-(hexyloxy)dodecyl]oxo}-6-oxohexyl)(4-hydroxybutyl)amino]hexanoate was obtained as a pale yellow liquid by column chromatography purification (600 mg, 59.3% yield).

[0319] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.15-3.95 (m, 4H), 3.58-3.48 (m, 4H), 3.47-3.39 (m, 4H), 2.50-2.47 (m, 4H) ),2.37-2.30(m,4H),1.69-1.59(m,7H),1.58-1.44(m,11H),1.37-1.13(m,52H),0.91-0.85(m,12H)ppm.

[0320] LCMS: RT=2.225, m / z 854.7[M+H]+.

[0321] Preparation Example 8 Preparation of Compound 12

[0322] Step 1: Synthesis of 2-(hexyloxy)dodecyl 6-[(6-{[2-(hexyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester

[0323] Reaction:

[0324]

[0325] Material ratio:

[0326] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Hexyloxy)dodecyl 6-bromohexyl ester 463 2.0 eq 1.00g 2.16 Ethanolamine 61 0.98eq 64.6mg 1.06 Potassium carbonate 138 7.0eq 1.04g 7.55 Potassium iodide 166 2.3 eq 412mg 2.48 Acetonitrile - - 8mL - Tetrahydrofuran - - 2mL -

[0327] Operation process:

[0328] 2-(Hexyloxy)dodecyl 6-bromohexyl ester (1.00 g, 1.06 mmol) was dissolved in acetonitrile (8 mL), and ethanolamine (64.6 mg, 1.06 mmol), potassium carbonate (1.04 g, 7.55 mmol), potassium iodide (412 mg, 2.48 mmol), tetrahydrofuran (2 mL) were added in sequence, and finally the mixture was stirred under nitrogen protection. The mixture was reacted at 70°C for 12 hours, and the reaction was completed after monitoring by TLC. The mixture was filtered and concentrated. Purification by column chromatography Obtained pale yellow liquid 2-(hexyloxy ) dodecanedioyl 6-[(6-{[2-(hexyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester (590 mg, 60.2% yield).

[0329] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.15-4.10 (m, 2H), 4.07-4.00 (m, 2H), 3.61-3.51 (m, 4H), 3.47-3.38 (m, 4H), 2.62 (t, J = 5. 2Hz,2H),2.54-2.46(m,4H),2.34(t,J=7.4Hz,4H),1.68-1.62(m,4H),1.59-1.45(m,12H),1.40-1.20(m,48H),0.89(dt,J 1 =2.6,J 2 =6.8Hz,12H)ppm.

[0330] LCMS:RT=2.266,m / z 826.7[M+H] + .

[0331] Preparation Example 9 Preparation of Compound 13

[0332] Step 1: Synthesis of heptadecan-9-yl 8-[(5-{[2-(hexyloxy)dodecyl]oxy}-5-oxypentyl)(2-hydroxyethyl)amino]octyl ester

[0333] Reaction:

[0334]

[0335] Material ratio:

[0336]

[0337] Operation process:

[0338] 2-(Hexyloxy) dodecyl 6-bromohexyl ester (700 mg, 1.56 mmol) was dissolved in acetonitrile (12 mL), and heptadecan-9-yl 8-[(2-hydroxyethyl) amino] octyl ester (688 mg, 1.56 mmol), potassium carbonate (753 mg, 5.45 mmol), potassium iodide (310 mg, 1.87 mmol), tetrahydrofuran (2 mL) were added in sequence, and finally reacted at 70 ° C for 12 hours under nitrogen protection, and the reaction was completed by TLC monitoring, filtered, and concentrated. The light yellow liquid heptadecan-9-yl 8-[(5-{[2-(hexyloxy) dodecyl] oxo}-5-oxyidene pentyl)(2-hydroxyethyl) amino] octyl ester (580 mg, 41.8% yield) was obtained by column chromatography purification.

[0339] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.87 (quin, J = 6.2Hz, 1H), 4.17-4.11 (m, 1H), 4.06-4.00 (m, 1H), 3.62 (t, J = 5.2Hz, 2H), 3.55 (td, J 1 =6.4,J 2 =9.2Hz,1H),3.46-3.39(m,2H),2.68(br t,J=5.2Hz,2H),2.62-2.53(m,4H),2.36(t,J=7.0Hz,2H),2.28(t,J=7.4Hz,2H),1 .67-1.60(m,4H),1.58-1.43(m,12H),1.36-1.21(m,52H),0.96-0.82(m,12H)ppm.

[0340] LCMS: RT=2.377, m / z 832.6[M+Na] + .

[0341] Preparation Example 10 Preparation of Compound 15

[0342] Step 1: Synthesis of ({[2-(heptyloxy)dodecyl]oxy}methyl)benzene

[0343] Reaction:

[0344]

[0345] Material ratio:

[0346] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-2-ol 292 1.00 eq 1.10g 3.76 1-Bromoheptane 178 1.20eq 0.808g 4.51 Sodium Hydrogen 24 2.00eq 0.300g 7.52 1-Methyl-2-pyrrolidone - - 10.0mL -

[0347] Operation process:

[0348] 1-(Benzyloxy) dodecan-2-ol (1.10 g, 3.76 mmol) was dissolved in 1-methyl-2-pyrrolidone (10.0 mL), sodium hydrogen (300 mg, 7.52 mmol) was added under nitrogen protection at 0°C, and the reaction was carried out for 1 hour at 0°C. 1-bromoheptane (808 mg, 4.51 mmol) was added to the reaction solution, and the reaction was carried out at 20°C under nitrogen protection for 12 hours. TLC monitoring showed that new spots were generated in the reaction, 15 mL of water was added, and the mixture was extracted with ethyl acetate twice, 20.0 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[2-(heptyloxy) dodecanyl] oxy} methyl) benzene (0.70 g, 47.6% yield).

[0349] Step 2: Synthesis of 2-(heptyloxy)dodecan-1-ol

[0350] Reaction:

[0351]

[0352] Material ratio:

[0353] Material Name Molecular weight Feed ratio Feeding amount mmol ({[2-(Heptyloxy)dodecyl]oxy}methyl)benzene 390 1.0 eq 1.00g 2.56 Palladium on Carbon 105.9 0.1eq 0.272g 0.255 Methanol - - 10.0mL -

[0354] Operation process:

[0355] Dissolve ({[2-(Heptyloxy)dodecyl]oxy}methyl)benzene (1.00 g, 2.56 mmol) in methanol (10.0 mL), add palladium carbon (272 mg, 255 μmol), and react at 35 ° C, 35 Psi for 12 hours under hydrogen protection. The reaction is completed after TLC monitoring. The mixture is filtered and concentrated to obtain a colorless liquid 2-(heptyloxy)dodecan-1-ol (0.70 g, 90.9% yield).

[0356] Step 3: Synthesis of 2-(heptyloxy)dodecyl 6-bromohexyl ester

[0357] Reaction:

[0358]

[0359] Material ratio:

[0360]

[0361]

[0362] Operation process:

[0363] 2-(Heptyloxy) dodecan-1-ol (700 mg, 2.33 mmol), 6-bromohexanoic acid (636 mg, 3.26 mmol), dissolved in dichloromethane (7.00 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (803 mg, 4.19 mmol), 4-dimethylaminopyridine (56.9 mg, 465 μmol) were added in sequence, and finally reacted at 25 ° C for 12 hours under nitrogen protection. TLC monitoring showed that new spots were generated in the reaction, 10 mL of water was added, and dichloromethane was extracted twice, 10 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain colorless liquid 2-(heptyloxy) dodecan-6-bromohexyl ester (0.89 g, 80.0% yield).

[0364] Step 4: Synthesis of 2-(heptyloxy)dodecyl 6-[(6-{[2-(heptyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester

[0365] Reaction:

[0366]

[0367] Material ratio:

[0368] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Heptyloxy)dodecyl 6-bromohexyl ester 476 1.0eq 890mg 1.86 2-Aminoethane-1-ol 61.05 0.48eq 54.6mg 0.894 Potassium carbonate 138 3.5 eq 901mg 6.52 Potassium iodide 166 1.2 eq 371mg 2.24 Acetonitrile - - 9.00mL - Tetrahydrofuran - - 4.50mL -

[0369] Operation process:

[0370] 2-(Heptyloxy) dodecyl 6-bromohexyl ester (0.89 g, 1.86 mmol) was dissolved in acetonitrile (9.00 mL), and 2-aminoethane-1-ol (54.6 mg, 894 μmol), potassium carbonate (901. mg, 6.52 mmol), potassium iodide (371 mg, 2.24 mmol), and tetrahydrofuran (4.50 mL) were added in sequence. Finally, the mixture was reacted at 75°C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a colorless liquid 2-(heptyloxy) dodecyl 6-[(6-{[2-(heptyloxy) dodecyl] oxo}-6-oxo-ylidenehexyl)(2-hydroxyethyl) amino]hexyl ester (441 mg, 20.0% yield).

[0371] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.17-4.08 (m, 2H), 4.07-3.98 (m, 2H), 3.64-3.51 (m, 4H), 3.43 (td, J1 = 6.6, J2 = 9.0Hz, 4H), 2.62 (br t,J=4.8Hz,2H),2.50(br t,J=7.2Hz,4H),2.34(t,J=7.4Hz,4H),1.70-1.63(m,4H),1.55-1.43(m,12H),1.37-1.13(m,52H),0.96-0.73(m,12H)ppm.

[0372] LCMS:RT=2.336,m / z 854.8[M+H] + .

[0373] Preparation Example 11 Preparation of Compound 20

[0374] Step 1: Synthesis of ({[4-(heptyloxy)dodecyl]oxy}methyl)benzene

[0375] Reaction:

[0376]

[0377] Material ratio:

[0378] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-4-ol 292 1.0 eq 4.72g 16.1 1-Bromoheptane 130 1.2eq 3.47g 19.3 Sodium Hydrogen 24.0 2.0eq 1.29g 32.2 N,N-Dimethylformamide - - 50mL -

[0379] Operation process:

[0380] 1-(Benzyloxy) dodecan-4-ol (4.72 g, 16.1 mmol) was dissolved in N, N-dimethylformamide (50 mL), sodium hydrogen (1.29 g, 32.2 mmol) was added at 0°C, the mixture was reacted at 0°C for 1 hour, 1-bromoheptane (3.47 g, 19.3 mmol) was added, and the mixture was reacted at 25°C for 12 hours under nitrogen protection. TLC monitoring showed that new spots were generated, 500 mL of water was added, and the mixture was extracted with ethyl acetate twice, 200 mL each time, and the organic phase was concentrated. Purification by column chromatography gave a colorless liquid ({[4-(heptyloxy) dodecanyl] oxy} methyl) benzene (2.5 g, 39.6% yield).

[0381] Step 2: Synthesis of 4-(heptyloxy)dodecan-1-ol

[0382] Reaction:

[0383]

[0384] Material ratio:

[0385] Material Name Molecular weight Feed ratio Feeding amount mmol ({[4-(Heptyloxy)dodecyl]oxy}methyl)benzene 390 1.0 eq 2.50g 6.40 Palladium on Carbon 105 0.2eq 1.36g 1.28 Methanol - - 25mL -

[0386] Operation process:

[0387] Dissolve ({[4-(heptyloxy)dodecyl]oxy}methyl)benzene (2.50 g, 6.40 mmol) in methanol (25 mL), add palladium carbon (1.36 g, 1.28 mmol), react at 35°C, 40 Psi for 12 hours under hydrogen protection, monitor the completion of the reaction by TLC, filter, and concentrate. The crude product was purified by column chromatography to obtain colorless liquid 4-(heptyloxy)dodecane-1-ol (390 mg, 20.2% yield).

[0388] Step 3: Synthesis of 4-(heptyloxy)dodecyl 6-bromohexyl ester

[0389] Reaction:

[0390]

[0391] Material ratio:

[0392]

[0393]

[0394] Operation process:

[0395] 4-(Heptyloxy) dodecan-1-ol (390 mg, 1.30 mmol), 6-bromohexanoic acid (303 mg, 1.56 mmol), dissolved in dichloromethane (5 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (373 mg, 1.95 mmol), 4-dimethylaminopyridine (15.8 mg, 0.129 mmol) were added in sequence, and finally reacted at 25°C for 12 hours under nitrogen protection. TLC monitored the reaction and new spots were generated. 50 mL of water was added, and dichloromethane was extracted twice, 50 mL each time, and the organic phase was concentrated. Purification by column chromatography gave a colorless liquid 4-(heptyloxy) dodecan-6-bromohexyl ester (540 mg, 87.1% yield).

[0396] Step 4: Synthesis of 4-(heptyloxy)dodecyl 6-[(6-{[4-(heptyloxy)dodecyl]oxy}-6-oxyylidenehexyl)(2-hydroxyethyl)amino]hexyl ester

[0397] Reaction:

[0398]

[0399] Material ratio:

[0400] Material Name Molecular weight Feed ratio Feeding amount mmol 4-(Heptyloxy)dodecyl 6-bromohexyl ester 477 1.0eq 540mg 1.13 2-Aminoethane-1-ol 61.0 0.45eq 31.0mg 0.508 Potassium carbonate 138 3.0 eq 468mg 3.39 Potassium iodide 166 1.5 eq 281mg 1.70 Acetonitrile - - 4mL - Tetrahydrofuran - - 2mL -

[0401] Operation process:

[0402] 4-(Heptyloxy) dodecyl 6-bromohexyl ester (540 mg, 1.13 mmol) was dissolved in acetonitrile (4.0 mL), and 2-aminoethane-1-ol (31.0 mg, 0.508 mmol), potassium carbonate (468 mg, 3.39 mmol), potassium iodide (281 mg, 1.70 mmol), and tetrahydrofuran (2 mL) were added in sequence. Finally, the mixture was reacted at 80° C. for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. 4-(Heptyloxy) dodecyl 6-[(6-{[4-(heptyloxy) dodecyl] oxo}-6-oxo-ylidenehexyl)(2-hydroxyethyl) amino]hexyl ester (130 mg, 11.7% yield) was obtained by column chromatography purification.

[0403] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.75 (s, 3H), 4.09 (t, J = 6.8Hz, 4H), 3.61 (brs, 2H), 3.47-3.36 (m, 4H), 3.24 (br t, J = 5.6Hz, 2H), 2.67 (br s, 2H), 2.55 (br s,3H),2.33(t,J=7.6Hz,4H),1.79-1.71(m,3H),1.58-1.47(m,18H),1.40-1.23(m,46H),0.90(t,J=6.8Hz,12H).

[0404] LCMS:RT=2.278,m / z 854.7[M+H] + .

[0405] Preparation Example 12 Preparation of Compound 21

[0406] Step 1: Synthesis of ({[2-(pentyloxy)dodecyl]oxy}methyl)benzene

[0407] Reaction:

[0408]

[0409] Material ratio:

[0410] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-2-ol 292 1.0 eq 2.00g 6.84 n-Pentyl bromide 151 1.3eq 1.34g 8.89 Sodium hydride (60%) 24 2.5 eq 684mg 17.1 N,N-Dimethylformamide - - 20mL -

[0411] Operation process:

[0412] 1-(Benzyloxy) dodecan-2-ol (2.00 g, 6.84 mmol) and sodium hydride (684 mg, 17.1 mmol, 60%) were dissolved in N, N-dimethylformamide (20 mL), stirred at 25°C for 0.5 hours under nitrogen protection, and n-pentane bromide (1.34 g, 8.89 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was carried out at 50°C for 6 hours under nitrogen protection, and the reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[2-(pentyloxy) dodecyl] oxy} methyl) benzene (1.20 g, 48.4% yield).

[0413] Step 2: Synthesis of 2-(pentyloxy)dodecan-1-ol

[0414] Reaction:

[0415]

[0416] Material ratio:

[0417] Material Name Molecular weight Feed ratio Feeding amount mmol ({[2-(Pentyloxy)dodecyl]oxy}methyl)benzene 362 1.0 eq 1.20g 3.31 Palladium / Carbon 106 0.10 eq 352mg 331 μmol hydrogen - (40Psi) - - Methanol - - 12mL -

[0418] Operation process:

[0419] Dissolve ({[2-(pentyloxy)dodecyl]oxy}methyl)benzene (1.20 g, 3.31 mmol) in methanol (12 mL), add palladium / carbon (352 mg, 331 umol) under nitrogen atmosphere, and finally react at 35°C for 12 hours under hydrogen (40 Psi) atmosphere. The reaction is completed by TLC monitoring, and the reaction liquid is filtered and concentrated. The crude product is purified by column chromatography to obtain colorless liquid 2-(pentyloxy)dodecan-1-ol (880 mg, 97.6% yield).

[0420] Step 3: Synthesis of 2-(pentyloxy)dodecyl 6-bromohexyl ester

[0421] Reaction:

[0422]

[0423] Material ratio:

[0424]

[0425]

[0426] Operation process:

[0427] 2-(Pentyloxy) dodecan-1-ol (880 mg, 3.23 mmol) was dissolved in dichloromethane (10 mL), and 6-bromohexanoic acid (820 mg, 4.20 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (929 mg, 4.84 mmol), and 4-dimethylaminopyridine (39.5 mg, 323 umol) were added in sequence. Finally, the mixture was reacted at 25°C for 12 hours under nitrogen protection. The reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain colorless liquid 2-(pentyloxy) dodecan-6-bromohexyl ester (1.10 g, 75.8% yield).

[0428] Step 4: Synthesis of 2-(pentyloxy)dodecyl 6-[(2-hydroxyethyl)(6-oxyylidene-6-{[2-(pentyloxy)dodecyl]oxy}hexyl)amino]hexyl ester

[0429] Reaction:

[0430]

[0431] Material ratio:

[0432] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Pentyloxy)dodecyl 6-bromohexyl ester 448 2.0eq 700mg 1.56 Ethanolamine 61 1.0eq 47.6mg 779 μmol Potassium carbonate 138 7.0 eq 753mg 5.45 Potassium iodide 166 2.2 eq 284mg 1.71 Acetonitrile - - 8mL - Tetrahydrofuran - - 2mL -

[0433] Operation process:

[0434] 2-(Pentyloxy) dodecyl 6-bromohexyl ester (700 mg, 1.56 mmol) was dissolved in acetonitrile (8 mL), and ethanolamine (47.6 mg, 779 umol), potassium carbonate (753 mg, 5.45 mmol), potassium iodide (284 mg, 1.71 mmol), tetrahydrofuran (2 mL) were added in sequence, and finally reacted at 80°C for 12 hours under nitrogen protection, and the reaction was completed after TLC monitoring, filtration, and concentration. Column chromatography was used for purification to obtain a light yellow liquid 2-(pentyloxy) dodecyl 6-[(2-hydroxyethyl)(6-oxyylidene-6-{[2-(pentyloxy) dodecyl] oxy} hexyl) amino] hexyl ester (480 mg, 72.5% yield).

[0435] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.15-4.10 (m, 2H), 4.07-4.00 (m, 2H), 3.58-3.52 (m, 4H), 3.43 (td, J 1 =6.6J 2 =,9.0Hz,4H),2.60(t,J=5.2Hz,2H),2.51-2.46(m,4H),2.34(t,J=7.4Hz,4H),1.68-1.61( m,4H),1.59-1.53(m,4H),1.53-1.41(m,10H),1.36-1.24(m,42H),0.95-0.83(m,12H)ppm.

[0436] LCMS: RT=2.130, m / z 820.6[M+Na] + .

[0437] Preparation Example 13 Preparation of Compound 5

[0438] Step 1: Synthesis of octyl 4-methylbenzene-1-sulfonic acid anion

[0439] Reaction:

[0440]

[0441] Material ratio:

[0442] Material Name Molecular weight Feed ratio Feeding amount mmol 7-Methyloctan-1-ol 144 1.00 eq 5.00g 31.6 Lithium Aluminum Tetrahydride 38 1.50eq 1.80g 47.4 Tetrahydrofuran - - 50mL -

[0443] Operation process:

[0444] Dissolve 7-methyloctan-1-ol (5.00 g, 31.6 mmol) in tetrahydrofuran (50 mL), add lithium aluminum tetrahydride (1.80 g, 47.4 mmol) at 0°C. Finally, react at 25°C for 2 hours under nitrogen protection, and monitor the completion of the reaction by TLC. Slowly quench with sodium sulfate decahydrate under nitrogen flow. The crude product is subjected to column chromatography to obtain colorless liquid 7-methyloctan-1-ol (4.20 g, 92.1% yield).

[0445] Step 2: Synthesis of octyl 4-methylbenzene-1-sulfonic acid anion

[0446] Reaction:

[0447]

[0448] Material ratio:

[0449] Material Name Molecular weight Feed ratio Feeding amount mmol 7-Methyloctan-1-ol 144 1.00 eq 3.60g 24.9 4-Methylbenzenesulfonyl chloride 190 1.50eq 7.14g 37.4 Triethylamine 101 3.00eq 7.58g 74.8 4-Dimethylaminopyridine 122 0.10 eq 305mg 2.05 Dichloromethane - - 40.0mL -

[0450] Operation process:

[0451] Dissolve 7-methyloctan-1-ol (3.60 g, 24.9 mmol) in dichloromethane (40.0 mL), add triethylamine (7.58 g, 37.4 mmol), 4-dimethylaminopyridine (305 mg, 2.05 mmol) in turn, and add 4-methylbenzenesulfonyl chloride (7.14 g, 37.4 mmol) at 0°C. Finally, react at 25°C for 2 hours under nitrogen protection, and the reaction is complete after TLC monitoring. Wash with water, extract, and concentrate. The crude product is subjected to column chromatography to obtain a colorless liquid 7-methyloctyl 4-methylbenzenesulfonic acid anion (6.60 g, 88.6% yield).

[0452] Step 3: Synthesis of 2-(7-methyloctaoxy)dodecyloxymethylbenzene

[0453] Reaction:

[0454]

[0455] Material ratio:

[0456]

[0457]

[0458] Operation process:

[0459] 1-(Benzyloxy) dodecan-2-ol (5.50 g, 15.1 mmol) and sodium hydride (1.81 g, 45.1 mmol, 60%) were dissolved in N, N-dimethylformamide (50.0 mL), stirred at 25°C for 0.5 hours under nitrogen protection, and 7-methyloctyl 4-methylbenzenesulfonic acid anion (5.39 g, 18.1 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was carried out at 50°C for 6 hours under nitrogen protection, and the reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain colorless liquid 2-(7-methyloctaoxy) dodecanoxymethylbenzene (3.70 g, 57.7% yield).

[0460] Step 4: Synthesis of 2-(7-methyloctaoxo)dodecan-1-ol

[0461] Reaction:

[0462]

[0463] Material ratio:

[0464] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(7-Methyloctaoxy)dodecyloxymethylbenzene 418 1.00 eq 3.30g 7.88 Palladium / Carbon 106 0.20eq 1.68g 1.58 hydrogen - (40Psi) - - Methanol - - 10.0mL -

[0465] Operation process:

[0466] Dissolve 2-(7-methyloctaoxy)dodecyloxymethylbenzene (3.30 g, 7.88 mmol) in methanol (10.0 mL), add palladium / carbon (1.68 g, 1.58 mmol) under argon atmosphere, and finally react at 35°C for 12 hours under hydrogen (40 Psi) atmosphere. The reaction is completed by TLC monitoring, and the reaction liquid is filtered and concentrated. The crude product is spin-dried and subjected to column chromatography to obtain colorless liquid 2-(7-methyloctaoxy)dodecane-1-ol (2.27 g, 87.6% yield).

[0467] Step 5: Synthesis of 2-(7-methyloctaoxo)dodecyl 6-bromohexyl ester

[0468] Reaction:

[0469]

[0470] Material ratio:

[0471]

[0472] Operation process:

[0473] Dissolve 2-(7-methyloctaoxo)dodecan-1-ol (2.27.g, 6.91mmol) in dichloromethane (10.0mL), add 6-bromohexane-1-acid (2.02g, 10.4mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.99g, 10.4mmol), 4-dimethylaminopyridine (84.4mg, 690μmol) in sequence, and finally react at 25℃ for 12 hours under nitrogen protection. The reaction is completed by TLC monitoring, and the reaction solution is concentrated. The crude product is subjected to column chromatography to obtain colorless liquid 2-(7-methyloctaoxo)dodecyl 6-bromohexyl ester (3.30g, 94.5% yield).

[0474] Step 6: Synthesis of 2-(7-methyloctaoxy)dodecyl 6-[2-hydroxyethyl-[6-[2-(7-methyloctaoxy)dodecyloxy]-6-oxyylidene-hexyl]amino]hexyl ester

[0475] Reaction:

[0476]

[0477] Material ratio:

[0478]

[0479] Operation process:

[0480] 2-(7-methyloctaoxo)dodecyl 6-bromohexyl ester (1.50 g, 2.97 mmol) was dissolved in acetonitrile (3.00 mL), and ethanolamine (81.6 mg, 1.34 μmol), potassium carbonate (615 mg, 4.45 mmol), potassium iodide (370 mg, 2.23 mmol), and tetrahydrofuran (1.00 mL) were added in sequence. Finally, the mixture was reacted at 75 ° C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a colorless liquid 2-(7-methyloctaoxo)dodecyl 6-[2-hydroxyethyl-[6-[2-(7-methyloctaoxo)dodecyloxy]-6-oxyylidene-hexyl]amino]hexyl ester (870 mg, 58.6% yield).

[0481] 1H NMR(400MHz,CHLOROFORM-d)δppm 4.14(d,J=4.25Hz,1H)4.11(d,J=4.00Hz,1H)3.98-4.08(m,2H)3.73(br t,J=4.75Hz,2H)3.51-3.60(m,2H)3.34-3.50(m,5H)2.82(br s,2H)2.61-2.76(m,4H)2.35(t,J=7.38Hz,6H)2.05(s,1H)1.60-1.74(m,9H) 1.41-1.58(m,13H)1.24-1.40(m,48H)1.09-1.20(m,6H)0.80-0.92(m,19H).

[0482] LCMS: RT=1.764, m / z=911.3[M+H] + .

[0483] Preparation Example 14 Preparation of Compound 8

[0484] Step 1: Synthesis of octyl 4-methylbenzene-1-sulfonic acid anion

[0485] Reaction:

[0486]

[0487] Material ratio:

[0488]

[0489]

[0490] Operation process:

[0491] Dissolve n-octanol (4.00 g, 30.7 mmol) in dichloromethane (40 mL), add 4-methylbenzenesulfonyl chloride (8.78 g, 46.1 mmol) at 0°C. Finally, react at 25°C for 2 hours under nitrogen protection, and monitor the completion of the reaction by TLC. Wash with water, extract, and concentrate. The crude product is subjected to column chromatography to obtain a colorless liquid octyl 4-methylbenzene-1-sulfonic acid anion (5.80 g, 66.4% yield).

[0492] Step 2: Synthesis of 1-(benzyloxy)dodecan-3-ol

[0493] Reaction:

[0494]

[0495] Material ratio:

[0496] Material Name Molecular weight Feed ratio Feeding amount mmol 3-(Benzyloxy)propanal 164 1.0 eq 4.40g 26.8 Heptylmagnesium bromide 230 1.2eq 7.44g 32.2 Tetrahydrofuran - - 50mL -

[0497] Operation process:

[0498] 3-(Benzyloxy)propanal (4.40 g, 26.8 mmol) was dissolved in tetrahydrofuran (50 mL), and a tetrahydrofuran solution of heptylmagnesium bromide (7.44 g, 32.2 mmol, 0.5 M) was slowly added dropwise at 0°C, and the mixture was reacted at 25°C for 12 hours under nitrogen protection. The reaction was completed by TLC monitoring, and the reaction solution was concentrated. The crude product was purified by column chromatography to obtain a colorless liquid 1-(benzyloxy)dodecan-3-ol (4.90 g, 62.5% yield).

[0499] Step 3: Synthesis of ({[3-(octyloxy)dodecyl]oxy}methyl)benzene

[0500] Reaction:

[0501]

[0502] Material ratio:

[0503] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-3-ol 293 1.0 eq 2.00g 6.84 Octyl 4-methylbenzene-1-sulfonic acid anion 284 1.5eq 2.92g 10.3 Sodium hydride (60%) 24 3.0 eq 821mg 20.5 N,N-Dimethylformamide - - 20mL -

[0504] Operation process:

[0505] 1-(Benzyloxy) dodecan-3-ol (2.00 g, 6.84 mmol) and sodium hydride (821 mg, 20.5 mmol, 60%) were dissolved in N, N-dimethylformamide (20 mL), stirred at 25°C for 0.5 hours under nitrogen protection, and octyl 4-methylbenzene-1-sulfonic acid anion (2.92 g, 10.3 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was carried out at 50°C for 6 hours under nitrogen protection, and the reaction was completed after TLC monitoring. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[3-(octyloxy) dodecanyl] oxy} methyl) benzene (2.30 g, 83.1% yield).

[0506] Step 5: Synthesis of 3-octaoxododecyl 6-bromohexyl ester

[0507] Reaction:

[0508]

[0509] Material ratio:

[0510]

[0511] Operation process:

[0512] Dissolve 3-octaoxododecan-1-ol (1.70 g, 5.40 mmol) in dichloromethane (10.0 mL), add 6-bromohexane-1-ol (1.58 g, 8.77 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.55 g, 8.11 mmol), 4-dimethylaminopyridine (66.0 mg, 540 μmol) in sequence, and finally react at 25 ° C for 12 hours under nitrogen protection. The reaction is completed by TLC monitoring, and the reaction solution is concentrated. The crude product is subjected to column chromatography to obtain a colorless liquid 3-octaoxododecan-6-bromohexyl ester (2.30 g, 86.6% yield).

[0513] Step 5: Synthesis of 3-octaoxododecyl 6-[2-hydroxyethyl-[6-(3-octaoxododecyloxy)-6-oxyylidene-hexyl]amino]hexyl ester

[0514] Reaction:

[0515]

[0516] Material ratio:

[0517] Material Name Molecular weight Feed ratio Feeding amount mmol 3-Octoxododecyl 6-bromohexyl ester 491 2.00 eq 1.60g 3.25 Ethanolamine 61 0.98eq 99.4mg 1.63 Potassium carbonate 138 3.00eq 0.675g 4.88 Potassium iodide 166 1.50 eq 405mg 2.44 Acetonitrile - - 10.0mL - Tetrahydrofuran - - 5.00mL -

[0518] Operation process:

[0519] 3-octaoxo-dodecyl 6-bromohexyl ester (1.60 g, 3.25 mmol) was dissolved in acetonitrile (10.0 mL), and ethanolamine (99.4 mg, 1.63 mmol), potassium carbonate (675 mg, 4.88 mmol), potassium iodide (405 mg, 2.44 mmol), and tetrahydrofuran (5.00 mL) were added in sequence. Finally, the mixture was reacted at 75 ° C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave a colorless liquid 3-octaoxo-dodecyl 6-[2-hydroxyethyl-[6-(3-octaoxo-dodecyloxy)-6-oxyylidene-hexyl]amino]hexyl ester (570 mg, 37.7% yield).

[0520] 1 H NMR (400MHz, CHLOROFORM-d) δppm 4.17 (td, J=6.69, 2.00Hz, 4H) 3.95 (br s,2H)3.25-3.63(m,7H)2.89-3.16(m,6H)2.33(t,J=7.25Hz,5H)1.74-1.88(m, 9H)1.68(dt,J=15.23,7.46Hz,6H)1.49-1.59(m,7H)1.37-1.48(m,8H)1.27(br s,43H)0.89(t,J=6.75Hz,12H).

[0521] LCMS: RT=1.785, m / z=882.4.[M+H] + .

[0522] Preparation Example 15 Preparation of Compound 9

[0523] Step 1: Synthesis of 2,3-di(octyloxy)propyl 5-bromopentyl ester

[0524] Reaction:

[0525]

[0526] Material ratio:

[0527]

[0528] Operation process:

[0529] Dissolve 1,3-di(octyloxy)propane-2-ol (500 mg, 1.58 mmol) in dichloromethane (5 mL), add 5-bromovaleric acid (343 mg, 1.90 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (454 mg, 2.37 mmol), 4-dimethylaminopyridine (19.3 mg, 0.158 mmol) in sequence, and finally react at 25°C for 12 hours under nitrogen protection. After TLC monitoring, the reaction is complete, 50 mL of water is added, and dichloromethane is extracted twice, 50 mL each time, and the organic phase is concentrated. The crude product is subjected to column chromatography to obtain a colorless liquid 2,3-di(octyloxy)propyl 5-bromopentyl ester (750 mg, 99.0% yield).

[0530] Step 2: Synthesis of heptadecan-9-yl 8-({5-[2,3-di(octyloxy)propoxy]-5-oxypentylene}(2-hydroxyethyl)amino)octyl ester

[0531] Reaction:

[0532]

[0533] Material ratio:

[0534]

[0535]

[0536] Operation process:

[0537] Dissolve 2,3-di(octyloxy)propyl 5-bromopentyl ester (550 mg, 1.15 mmol) in acetonitrile (5 mL), add heptadecan-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (422 mg, 0.955 mmol), potassium carbonate (462 mg, 3.35 mmol), potassium iodide (190 mg, 1.15 mmol), tetrahydrofuran (2 mL) in sequence, and finally react at 75°C for 12 hours under nitrogen protection. After TLC monitoring, the reaction is completed, filtered, and concentrated. Purification by column chromatography gives a yellow liquid heptadecan-9-yl 8-({5-[2,3-di(octyloxy)propoxy]-5-oxypentyl}(2-hydroxyethyl)amino)octyl ester (120 mg, 14.0% yield).

[0538] 1 H NMR (400MHz, CHLOROFORM-d) δ = 4.93-4.81 (m, 1H), 4.24 (dd, J = 4.0, 11.6Hz, 1H), 4.11 (dd, J=5.6,11.6Hz,1H),3.66-3.61(m,1H),3.56(t,J=6.8Hz,4H),3.52-3.39(m,4H),2.62(br s,2H),2.50(br s,4H),2.36(t,J=7.2Hz,2H),2.28(t,J=7.6Hz,2H),1.72-1.54(m,15H),1.37-1.17(m,52H),0.89(t,J=6.8Hz,12H).

[0539] LCMS:RT=2.118,m / z 886.7[M+H] + .

[0540] Preparation Example 16 Preparation of Compound 14

[0541] Step 1: Synthesis of ({[5-(hexyloxy)dodecyl]oxy}methyl)benzene

[0542] Reaction:

[0543]

[0544] Material ratio:

[0545] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-5-ol 292 1.00 eq 1.20g 4.10 1-Bromohexane 164 1.20eq 0.328g 8.21 Sodium Hydrogen 24 2.00eq 0.880g 5.33 1-Methyl-2-pyrrolidone - - 12.0mL -

[0546] Operation process:

[0547] 1-(Benzyloxy) dodecan-5-ol (1.20 g, 4.10 mmol) was dissolved in 1-methyl-2-pyrrolidone (12.0 mL), sodium hydrogen (328 mg, 8.21 mmol) was added under nitrogen protection at 0°C, and the reaction was carried out at 0°C for 1 hour. 1-bromohexane (880 mg, 5.33 mmol) was added to the reaction solution, and the reaction was carried out at 25°C under nitrogen protection for 12 hours. TLC monitoring showed that new spots were generated in the reaction, 15 mL of water was added, and the mixture was extracted with ethyl acetate twice, 20.0 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[5-(hexyloxy) dodecanyl] oxy} methyl) benzene (0.60 g, 38.3% yield).

[0548] Step 2: Synthesis of 5-(hexyloxy)dodecan-1-ol

[0549] Reaction:

[0550]

[0551] Material ratio:

[0552]

[0553] Operation process:

[0554] Dissolve ({[5-(Hexyloxy)dodecyl]oxy}methyl)benzene (600 mg, 1.59 mmol) in methanol (6.00 mL), add palladium carbon (339 mg, 318 μmol), and react at 35°C, 35 Psi for 12 hours under hydrogen protection. The reaction is completed after TLC monitoring. The mixture is filtered and concentrated to obtain colorless liquid 5-(hexyloxy)dodecan-1-ol (0.10 g, 21.9% yield).

[0555] Step 3: Synthesis of 5-(hexyloxy)dodecyl 5-bromopentyl ester

[0556] Reaction:

[0557]

[0558] Material ratio:

[0559]

[0560] Operation process:

[0561] Combine the two batches of materials, dissolve 5-(hexyloxy) dodecan-1-ol (420 mg, 1.47 mmol) and 5-bromovaleric acid (345 mg, 1.91 mmol) in dichloromethane (5.00 mL), add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (505 mg, 2.64 mmol) and 4-dimethylaminopyridine (35.8 mg, 293 μmol) in sequence, and finally react at 25°C for 12 hours under nitrogen protection. TLC monitors the reaction to see new spots, add 10.0 mL of water, extract with dichloromethane twice, 10 mL each time, and concentrate the organic phase. The crude product is subjected to column chromatography to obtain colorless liquid 5-(hexyloxy) dodecan-5-bromopentyl ester (0.418 g, 63.4% yield).

[0562] Step 4: Synthesis of heptadecan-9-yl 8-[(5-{[5-(hexyloxy)dodecyl]oxy}-5-oxyylidenepentyl)(2-hydroxyethyl)amino]octyl ester

[0563] Reaction:

[0564]

[0565] Material ratio:

[0566]

[0567]

[0568] Operation process:

[0569] 5-(Hexyloxy) dodecyl 5-bromopentyl ester (368 mg, 818 μmol) was dissolved in acetonitrile (6.00 mL), and heptadecan-9-yl 8-[(2-hydroxyethyl)amino]octyl ester (361 mg, 818 μmol), potassium carbonate (396 mg, 2.87 mmol), potassium iodide (163 mg, 982 μmol), tetrahydrofuran (3.00 mL) were added in sequence, and finally reacted at 75°C for 12 hours under nitrogen protection. The reaction was monitored by TLC and concentrated. Purification by column chromatography gave a colorless liquid heptadecan-9-yl 8-[(5-{[5-(hexyloxy) dodecyl] oxy}-5-oxyylidenepentyl)(2-hydroxyethyl)amino]octyl ester (438 mg, 60.6% yield).

[0570] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.87 (t, J = 6.2Hz, 1H), 4.07 (t, J = 6.8Hz, 2H), 3.57 (t, J = 5.4Hz, 2H), 3.40 (dt, J1 = 4.4, J2 = 6.6Hz, 2H), 3.19 (br d,J=5.4Hz,1H),2.62(br t,J=5.2Hz,2H),2.57-2.43(m,4H),2.30(td,J1=7.5,J2=17.0Hz,4H),1.71- 1.59(m,8H),1.54-1.45(m,12H),1.38-1.20(m,48H),1.00-0.73(m,12H)ppm.

[0571] LCMS:RT=2.257,m / z 810.7[M+1] + .

[0572] Preparation Example 17 Preparation of Compound 22

[0573] Step 1: Synthesis of 7-butoxydodecyloxymethylbenzene

[0574] Reaction:

[0575]

[0576] Material ratio:

[0577] Material Name Molecular weight Feed ratio Feeding amount mmol 12-Benzyloxydodecane-6-ol 292 1.00 eq 1.60g 5.47 1-Bromobutane 137 1.30 eq 974mg 7.11 Sodium hydride (60%) 24 2.50 eq 547mg 13.7 1-Methyl-2-pyrrolidone - - 20.0mL -

[0578] Operation process:

[0579] 12-Benzyloxydodecan-6-ol (1.60 g, 5.47 mmol) and sodium hydride (574 mg, 13.7 mmol, 60%) were dissolved in 1-methyl-2-pyrrolidone (20 mL), stirred at 25°C for 0.5 hours under nitrogen protection, and 1-bromobutane (974 mg, 7.11 mmol) was slowly added dropwise to the reaction mixture. Finally, the reaction was carried out at 50°C for 6 hours under nitrogen protection. The reaction was completed by TLC monitoring, and the reaction solution was poured into ice water and extracted with EtOAC. The reaction solution was concentrated, and the crude product was subjected to column chromatography to obtain colorless liquid 7-butoxydodecyloxymethylbenzene (1.00 g, 52.4% yield).

[0580] Step 2: Synthesis of 2-(7-methyloctaoxo)dodecan-1-ol

[0581] Reaction:

[0582]

[0583] Material ratio:

[0584] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(7-Methyloctaoxy)dodecyloxymethylbenzene 348.6 1.0 eq 3.30g 7.88 Palladium / Carbon 106 0.20eq 1.68g 1.58 hydrogen - (40Psi) - - Methanol - 10.0mL -

[0585] Operation process:

[0586] Dissolve 2-(7-methyloctaoxy)dodecyloxymethylbenzene (1.00 g, 2.87 mmol) in methanol (10.0 mL), add palladium / carbon (305 mg, 287 umol) under argon atmosphere, and finally react at 35°C for 12 hours under hydrogen (40 Psi) atmosphere. The reaction is completed by TLC monitoring, and the reaction liquid is filtered and concentrated. The crude product is spin-dried and subjected to column chromatography to obtain colorless liquid 2-(7-methyloctaoxy)dodecane-1-ol (700 mg, 94.7% yield).

[0587] Step 3: Synthesis of 7-butoxydodecyl 5-bromopentyl ester

[0588] Reaction:

[0589]

[0590] Material ratio:

[0591]

[0592] Operation process:

[0593] Dissolve 7-butoxydodecan-1-ol (700.mg, 2.71mmol) in dichloromethane (10.0mL), add 5-bromovaleric acid (637mg, 3.52mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (788mg, 4.06mmol), 4-dimethylaminopyridine (33.1mg, 270μmol) in sequence, and finally react at 25℃ for 12 hours under nitrogen protection. The reaction is completed by TLC monitoring, and the reaction solution is concentrated. The crude product is subjected to column chromatography to obtain colorless liquid 7-butoxydodecyl 5-bromopentyl ester (800mg, 70.8% yield).

[0594] Step 4: Synthesis of 1-octylnonyl 8-[[5-(7-butoxydodecyloxy)-5-oxyylidene-pentyl]-(2-hydroxyethyl)amino]octyl ester

[0595] Reaction:

[0596]

[0597] Material ratio:

[0598] Material Name Molecular weight Feed ratio Feeding amount mmol 7-Butoxydodecyl 5-bromopentyl ester 421 1.00eq 800mg 1.90 1-Octylnonyl 8-(2-hydroxyethylamino)octyl ester 441 1.00 eq 800mg 1.90 Potassium carbonate 138 3.50 eq 918mg 6.64 Potassium iodide 166 1.20 eq 378mg 2.28 Acetonitrile - - 5.00mL - Tetrahydrofuran - - 15.0mL -

[0599] Operation process:

[0600] 7-Butoxydodecyl 5-bromopentyl ester (800 mg, 1.90 mmol) was dissolved in acetonitrile (15.0 mL), and 1-octylnonyl 8-(2-hydroxyethylamino)octyl ester (800 mg, 1.90 μmol), potassium carbonate (918 mg, 6.64 mmol), potassium iodide (378 mg, 2.28 mmol), and tetrahydrofuran (5.00 mL) were added in sequence. Finally, the mixture was reacted at 75°C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. Purification by column chromatography gave colorless liquid 1-octylnonyl 8-[[5-(7-butoxydodecyloxy)-5-oxyylidene-pentyl]-(2-hydroxyethyl)amino]octyl ester (900 mg, 45.5% yield).

[0601] 1H NMR(400MHz,CHLOROFORM-d)δppm 4.87(t,J=6.19Hz,1H)4.07(t,J=6.75Hz,2H)3.58(t,J=5.25Hz,2H)3.41(t,J=6.63Hz,2H)3.19(br t,J=5.25Hz,1H)2.63(t,J=5.19Hz,2H)2.45-2.56(m,4H)2.30(dt,J=17 .60,7.39Hz,4H)1.78-1.90(m,1H)1.55-1.71(m,8H)0.85-0.97(m,11H).

[0602] LCMS: RT=1.764, m / z=911.3[M+H] + .

[0603] Preparation Example 18 Preparation of Compound 23

[0604] Step 1: Synthesis of ({[1,3-di(octyloxy)propane-2-yl]oxy}methyl)benzene

[0605] Reaction:

[0606]

[0607] Material ratio:

[0608] Material Name Molecular weight Feed ratio Feeding amount mmol 2-(Benzyloxy)propane-1,3-diol 182 1.0 eq 3.00g 16.4 1-Bromooctane 192 2.2eq 7.00g 36.2 Sodium Hydrogen 24 3.0eq 1.98g 49.3 N,N-Dimethylformamide - - 30mL -

[0609] Operation process:

[0610] 2-(Benzyloxy)propane-1,3-diol (3.00 g, 16.4 mmol) was dissolved in N,N-dimethylformamide (30 mL), sodium hydrogen (1.98 g, 49.3 mmol) was added under nitrogen protection at 25°C, and the reaction was continued for one hour at 25°C. 1-Bromooctane (7.00 g, 36.2 mmol) was added to the reaction solution, and the reaction was continued at 25°C under nitrogen protection for 12 hours. TLC monitoring showed that new spots were generated in the reaction, 300 mL of water was added, and the mixture was extracted twice with ethyl acetate, 100 mL each time, and the organic phase was concentrated. The crude product was subjected to column chromatography to obtain a colorless liquid ({[1,3-di(octyloxy)propane-2-yl]oxy}methyl)benzene (3.30 g, 49.2% yield).

[0611] Step 2: Synthesis of 1,3-di(octyloxy)propane-2-ol

[0612] Reaction:

[0613]

[0614] Material ratio:

[0615] Material Name Molecular weight Feed ratio Feeding amount mmol ({[1,3-Di(octyloxy)propan-2-yl]oxy}methyl)benzene 406 1.0 eq 3.30g 8.12 Palladium on Carbon 105 0.2eq 1.73g 1.62 Methanol - - 30mL -

[0616] Operation process:

[0617] Dissolve ({[1,3-di(octyloxy)propane-2-yl]oxy}methyl)benzene (3.30 g, 8.12 mmol) in methanol (30 mL), add palladium carbon (1.73 g, 1.62 mmol), react at 35°C, 40 Psi for 12 hours under hydrogen protection, monitor the reaction completion by TLC, filter, and concentrate. The crude product was purified by column chromatography to obtain colorless liquid 1,3-di(octyloxy)propane-2-ol (2.27 g, 88.3% yield).

[0618] Step 3: Synthesis of 3-(heptyloxy)-2-(octyloxy)propyl 6-bromohexyl ester

[0619] Reaction:

[0620]

[0621] Material ratio:

[0622]

[0623] Operation process:

[0624] Dissolve 1,3-di(octyloxy)propane-2-ol (1.00 g, 3.16 mmol) in dichloromethane (10 mL), add 6-bromohexanoic acid (739 mg, 3.79 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (908 mg, 4.74 mmol), 4-dimethylaminopyridine (38.6 mg, 0.316 mmol) in sequence, and finally react at 25°C for 12 hours under nitrogen protection. After TLC monitoring, the reaction is complete, 50 mL of water is added, and dichloromethane is extracted twice, 50 mL each time, and the organic phase is concentrated. The crude product is subjected to column chromatography to obtain a colorless liquid 3-(heptyloxy)-2-(octyloxy)propyl 6-bromohexyl ester (1.50 g, 96.1% yield).

[0625] Step 4: Synthesis of 2,3-di(octyloxy)propyl 6-({6-[2,3-di(octyloxy)propoxy]-6-oxyylidenehexyl}(2-hydroxyethyl)amino)hexyl ester

[0626] Reaction:

[0627]

[0628] Material ratio:

[0629] Material Name Molecular weight Feed ratio Feeding amount mmol 3-(Heptyloxy)-2-(octyloxy)propyl 6-bromohexyl ester 492 1.0 eq 1.00g 2.03 2-Aminoethane-1-ol 61.0 0.45eq 55.6mg 0.911 Potassium carbonate 138 3.0 eq 840mg 6.08 Potassium iodide 166 1.5 eq 504mg 3.04 Acetonitrile - - 10mL - Tetrahydrofuran - - 2mL -

[0630] Operation process:

[0631] 3-(Heptyloxy)-2-(octyloxy)propyl 6-bromohexyl ester (1.00 g, 2.03 mmol) was dissolved in acetonitrile (10 mL), and 2-aminoethane-1-ol (55.6 mg, 0.911 mmol), potassium carbonate (840 mg, 6.08 mmol), potassium iodide (504 mg, 3.04 mmol), and tetrahydrofuran (2 mL) were added in sequence. Finally, the mixture was reacted at 75°C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. The yellow liquid 2,3-di(octyloxy)propyl 6-({6-[2,3-di(octyloxy)propoxy]-6-oxyhexyl}(2-hydroxyethyl)amino)hexyl ester (580 mg, 30.2% yield) was obtained by column chromatography purification.

[0632] 1H NMR (400MHz, CHLOROFORM-d) δ = 4.23 (dd, J = 4.0, 11.6Hz, 2H), 4.10 (dd, J = 5.6, 11.6Hz, 2H), 3.65-3.54 (m, 8H), 3.50-3.41 (m, 8H), 2.65 (br s, 2H), 2.53 (br t,J=7.2Hz,4H),2.34(t,J=7.6Hz,4H),1.64(qd,J=7.6,15.2Hz,15H),1.40-1.21(m,46H),0.89(t,J=6.8Hz,12H).

[0633] LCMS:RT=2.118,m / z 886.7[M+H] + .

[0634] Preparation Example 19 Preparation of Compound 16

[0635] Step 1: Synthesis of 3-ethylheptan-1-ol

[0636] Reaction:

[0637]

[0638] Material ratio:

[0639] Material Name Molecular weight Feed ratio Feeding amount mmol 3-Ethylheptanoic acid 158 1.0eq 500mg 3.16 Lithium aluminum hydride 2.5M tetrahydrofuran solution 37.9 1.3eq 1.64mL 4.11 Tetrahydrofuran - - 10mL -

[0640] Operation process:

[0641] 3-Ethylheptanoic acid (500 mg, 3.16 mmol) was dissolved in tetrahydrofuran (10 mL), cooled to 0°C, and lithium aluminum hydride 2.5 M tetrahydrofuran solution (1.64 mL, 4.11 mmol) was added dropwise to the reaction solution at 0-5°C. Finally, the reaction was carried out at 25°C for 12 hours under nitrogen protection. After the reaction was completed, 10 mL of saturated ammonium chloride aqueous solution was added, and the mixture was extracted with ethyl acetate twice, 20 mL each time. The organic phase was dried and concentrated. Purification by column chromatography gave a colorless liquid 3-ethylheptane-1-ol (344 mg, 75.4% yield).

[0642] Step 2: Synthesis of 3-ethylheptyl 4-methylbenzene-1-sulfonic acid anion

[0643] Reaction:

[0644]

[0645] Material ratio:

[0646] Material Name Molecular weight Feed ratio Feeding amount mmol 3-Ethylheptan-1-ol 144 1.0eq 340mg 2.36 p-Toluenesulfonyl chloride 190 1.5 eq 674mg 3.54 4-Dimethylaminopyridine 122 0.1eq 28.7mg 0.235 Triethylamine 101 3.0 eq 715mg 7.07 Dichloromethane - - 4mL -

[0647] Operation process:

[0648] 3-Ethylheptane-1-ol (340 mg, 2.36 mmol) was dissolved in dichloromethane (4 mL), 4-dimethylaminopyridine (28.7 mg, 0.235 mmol), triethylamine (715 mg, 7.07 mmol), p-toluenesulfonyl chloride (674 mg, 3.54 mmol) were added to the reaction solution in sequence, and the mixture was reacted at 25°C for 12 hours under nitrogen protection. The reaction was completed after TLC monitoring, and the mixture was concentrated. The mixture was purified by column chromatography to obtain a colorless liquid 3-ethylheptyl 4-methylbenzene-1-sulfonic acid anion (557 mg, 79.1% yield).

[0649] Step 3: Synthesis of [({2-[(3-ethylheptyl)oxy]dodecyl}oxy)methyl]benzene

[0650] Reaction:

[0651]

[0652] Material ratio:

[0653]

[0654] Operation process:

[0655] 1-(Benzyloxy) dodecan-2-ol (544 mg, 1.86 mmol) was dissolved in N, N-dimethylformamide (6 mL), sodium hydride (223 mg, 5.59 mmol) was added at 25°C, and the reaction was continued at 25°C for 1 hour. 3-ethylheptyl 4-methylbenzene-1-sulfonic acid anion (556 mg, 1.86 mmol) was added, and the reaction was continued at 50°C for 12 hours. The reaction was monitored by TLC, and 5 mL of saturated ammonium chloride aqueous solution was added. The mixture was extracted with ethyl acetate twice, each time with 10 mL. The organic phase was dried and concentrated. The mixture was purified by column chromatography to obtain a colorless liquid [({2-[(3-ethylheptyl)oxy]dodecyl}oxy)methyl]benzene (610 mg, 78.2% yield).

[0656] Step 4: Synthesis of 2-[(3-ethylheptyl)oxy]undecane-1-ol

[0657] Reaction:

[0658]

[0659] Material ratio:

[0660]

[0661] Operation process:

[0662] Dissolve [({2-[(3-ethylheptyl)oxy]dodecyl}oxy)methyl]benzene (820 mg, 1.96 mmol) in methanol (10 mL), add palladium carbon (416 mg, 0.391 mmol), react at 35°C, 40 Psi for 12 hours under hydrogen protection, monitor the completion of the reaction by TLC, filter, and concentrate. Purify by column chromatography to obtain colorless liquid 2-[(3-ethylheptyl)oxy]dodecane-1-ol (455 mg, 70.7% yield).

[0663] Step 5: Synthesis of 2-[(3-ethylheptyl)oxy]dodecyl 6-bromohexyl ester

[0664] Reaction:

[0665]

[0666] Material ratio:

[0667]

[0668] Operation process:

[0669] 2-[(3-ethylheptyl)oxy]dodecan-1-ol (450 mg, 1.37 mmol), 6-bromohexanoic acid (320 mg, 1.64 mmol), dissolved in dichloromethane (6 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (393 mg, 2.05 mmol), 4-dimethylaminopyridine (33.5 mg, 0.273 mmol) were added in sequence, and finally reacted at 25°C for 12 hours under nitrogen protection. TLC monitored the reaction and new spots were generated. The reaction solution was concentrated. The crude product was subjected to column chromatography to obtain colorless liquid 2-[(3-ethylheptyl)oxy]dodecyl 6-bromohexyl ester (680 mg, 98.2% yield).

[0670] Step 6: Synthesis of 2-[(3-ethylheptyl)oxy]dodecyl 6-{[6-({2-[(3-ethylheptyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl](2-hydroxyethyl)amino}hexyl ester

[0671] Reaction:

[0672]

[0673] Material ratio:

[0674]

[0675] Operation process:

[0676] 2-[(3-ethylheptyl)oxy]dodecyl 6-bromohexyl ester (580 mg, 1.15 mmol) was dissolved in acetonitrile (4 mL), and 2-aminoethane-1-ol (31.5 mg, 0.516 mmol), potassium carbonate (475 mg, 3.44 mmol), potassium iodide (285 mg, 1.72 mmol), tetrahydrofuran (2 mL) were added in sequence, and finally reacted at 75° C. for 12 hours under nitrogen protection, and the reaction was completed after TLC monitoring, filtration, and concentration. Purification by column chromatography gave a colorless liquid 2-[(3-ethylheptyl)oxy]dodecyl 6-{[6-({2-[(3-ethylheptyl)oxy]dodecyl}oxy)-6-oxyylidenehexyl](2-hydroxyethyl)amino}hexyl ester (260 mg, 21.8% yield).

[0677] 1 H NMR (400MHz, CHLOROFORM-d)δ=4.17-4.10(m,2H),4.07-4.01(m,2H),3.62-3.53(m,4H),3.50-3.39(m,4H),2.62(br s,2H),2.50(br s,4H),2.34(t,J=7.6Hz,4H),1.70-1.61(m,6H),1.55-1.46(m,13H),1.38-1.21(m,52H),0.96-0.80(m,18H)ppm.

[0678] LCMS:RT=3.130,m / z 910.8[M+H] + .

[0679] Preparation Example 20 Preparation of Compound 24

[0680] Step 1: Synthesis of (4Z)-oct-4-en-1-ol

[0681] Reaction:

[0682]

[0683] Material ratio:

[0684] Material Name Molecular weight Feed ratio Feeding amount mmol (4Z)-Oct-4-enoic acid 142 1.00eq 693mg 4.88 Lithium aluminum hydride 37 1.30eq 2.54mL 6.30 Tetrahydrofuran - - 8.30mL -

[0685] Operation process:

[0686] Dissolve (4Z)-oct-4-enoic acid (693 mg, 4.88 mmol) in tetrahydrofuran (8.30 mL), add lithium aluminum hydride (2.5 M, 2.54 mL) at 0°C, react at 0°C for 2 hours, monitor the completion of the reaction by TLC, quench, extract twice with ethyl acetate, and concentrate the organic phase to obtain a colorless liquid (4Z)-oct-4-en-1-ol (575 mg, 91.9% yield).

[0687] Step 2: Synthesis of (4Z)-1-iodooct-4-ene

[0688] Reaction:

[0689]

[0690] Material ratio:

[0691] Material Name Molecular weight Feed ratio Feeding amount mmol (4Z)-Oct-4-en-1-ol 128 1.00 eq 475 mg 3.70 Imidazole 68 1.30 eq 327 mg 4.82 Triphenylphosphine 262 1.30 eq 1.26 g 4.82 Elemental iodine 253 1.30 eq 1.22 g 4.82 Acetonitrile 5.00 mL Tetrahydrofuran 10.0 mL

[0692] Operation process:

[0693] Dissolve (4Z)-oct-4-en-1-ol (475 mg, 3.70 mmol) in tetrahydrofuran (10.0 mL) and acetonitrile (5.00 mL), add triphenylphosphine (1.26 g, 4.82 mmol), imidazole (327.mg, 4.82 mmol) and elemental iodine (1.22 g, 4.82 mmol) at 0°C, react at 25°C under nitrogen atmosphere for 12 hours, monitor the reaction by TLC, concentrate and purify by column to obtain colorless liquid (4Z)-1-iodooct-4-ene (543 mg, 61.5% yield)

[0694] Step 3: Synthesis of [({2-[(4Z)-oct-4-ene-1-oxy]dodecyl}oxy)methyl]benzene

[0695] Reaction:

[0696]

[0697] Material ratio:

[0698] Material Name Molecular weight Feed ratio Feeding amount mmol 1-(Benzyloxy)dodecane-2-ol 292 1.00 eq 600 mg 2.05 (4Z)-1-iodooct-4-ene 238 1.00 eq 488 mg 2.05 Sodium Hydrogen 24 2.00 eq 164 mg 4.10 NN dimethylformamide - - 6.00 mL -

[0699] Operation process:

[0700] 1-(Benzyloxy)dodecan-2-ol (600 mg, 2.05 mmol) was dissolved in NN dimethylformamide (6.00 mL), sodium hydrogen (164 mg, 4.10 mmol) was added under nitrogen atmosphere, stirred at 25°C for 1 hour, and (488 mg, 2.05 mmol) was added under nitrogen protection at 25°C, and finally stirred at 60°C for 12 hours. TLC monitoring showed that new spots were generated in the reaction, and the organic phase was quenched, extracted and concentrated. Purification by column chromatography gave a colorless liquid [({2-[(4Z)-oct-4-ene-1-oxy] dodecyl}oxy)methyl]benzene (123 mg, 14.8% yield).

[0701] Step 4: Synthesis of 2-[(4Z)-oct-4-en-1-oxy]dodecan-1-ol

[0702] Reaction:

[0703]

[0704] Material ratio:

[0705]

[0706] Operation process:

[0707] Dissolve [({2-[(4Z)-oct-4-ene-1-oxy] dodecyl} oxy) methyl] benzene (100 mg, 248 μmol) in dichloromethane (2.00 mL), add boron trichloride (1M, 496 μL) at 0°C, and react at 0°C for 1 hour under nitrogen protection. TLC monitors the formation of new spots in the reaction. After adjusting the pH to neutral, extract with ethyl acetate, and concentrate the organic phase. Purify by column chromatography to obtain colorless liquid 2-[(4Z)-oct-4-ene-1-oxy] dodecan-1-ol (70.0 mg, 90.1% yield).

[0708] Step 5: Synthesis of 2-[(4Z)-oct-4-ene-1-oxy]dodecyl 6-bromohexyl ester

[0709] Reaction:

[0710]

[0711] Material ratio:

[0712]

[0713] Operation process:

[0714] 2-[(4Z)-oct-4-ene-1-oxy] dodecan-1-ol (80.0 mg, 255 μmol) and 6-bromohexanoic acid (69.9 mg, 358 μmol) were dissolved in dichloromethane (2.00 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (73.6 mg, 383 μmol) and 4-dimethylaminopyridine (3.13 mg, 25.6 μmol) were added in sequence. Finally, the mixture was reacted at 25°C for 12 hours under nitrogen protection. TLC monitored the formation of new spots in the reaction, and the organic phase was concentrated. Purification by column chromatography gave a colorless liquid 2-[(4Z)-oct-4-ene-1-oxy] dodecan-6-bromohexyl ester (86.0 mg, 68.6% yield).

[0715] Step 6: Synthesis of 2-[(4Z)-oct-4-ene-1-oxy]dodecyl 6-[(2-hydroxyethyl)[6-({2-[(4Z)-oct-4-ene-1-oxy]dodecyl}oxy)-6-oxyylidenehexyl]amino]hexyl ester

[0716] Reaction:

[0717]

[0718] Material ratio:

[0719]

[0720] Operation process:

[0721] 2-[(4Z)-oct-4-ene-1-oxy] dodecyl 6-bromohexyl ester (86.0 mg, 175 μmol) was dissolved in acetonitrile (2.00 mL), and 2-aminoethane-1-ol (5.15 mg, 84.3 μmol), potassium carbonate (84.9 mg, 614 μmol), potassium iodide (34.9 mg, 210 μmol), and tetrahydrofuran (1.00 mL) were added in sequence. Finally, the mixture was reacted at 75°C for 12 hours under nitrogen protection. The reaction was monitored by TLC, filtered, and concentrated. The colorless liquid 2-[(4Z)-oct-4-ene-1-oxy] dodecyl 6-[(2-hydroxyethyl)[6-({2-[(4Z)-oct-4-ene-1-oxy] dodecyl}oxy)-6-oxyylidenehexyl]amino]hexyl ester (47.8 mg, 29.4% yield) was obtained by column chromatography purification.

[0722] 1 H NMR(400MHz,CHLOROFORM-d)δ=5.48-5.29(m,4H),4.18-4.11(m,2H),4.04(dd,J 1 =5.8,J 2 =11.4Hz,4H),3.56(td,J1 =6.6,J 2 =9.2Hz,2H),3.49-3.40(m,4H),3.21-3.05(m,5H),2.38(t,J=7.2Hz,4H),2.15-2.07(m,4H),2.05- 1.97(m,4H),1.95-1.84(m,4H),1.72-1.60(m,8H),1.45-1.21(m,46H),0.90(q,J=7.2Hz,12H)ppm.

[0723] LCMS:RT=2.710,m / z 878.8[M+H] + .

[0724] Example 1 Preparation and Detection of Lipid Nanoparticles (LNP)

[0725] 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., item number: S01005), cholesterol (purchased from Nippon Seika Co., Ltd., item number: O01001) and dimyristoylglycerol-polyethylene glycol 2000 (DMG-PEG2000, purchased from Guobang Pharmaceutical, item number: 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 an ethanol solution of mixed lipids, wherein the total lipid concentration is 12.5mM (the measurement unit "M" in the present application refers to mol / L). The firefly luciferase (Fluc) mRNA was diluted in a 25 mM sodium acetate buffer at pH 5.0 to obtain an mRNA solution. By using a microfluidic device, the flow rate was controlled to be 12 mL / min, the volume ratio of the ethanol solution of the mixed lipids to the mRNA solution obtained in the previous step was controlled to be 1:3, and lipid nanoparticles were prepared according to the nitrogen-phosphorus ratio of the ionizable lipids to the mRNA of 3 to 15:1. The ethanol was removed by dialysis in 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, item number: SLGPR33RB), and ultrafiltration concentration (manufacturer: Amicon-Ultra, molecular weight cutoff: 10 kDa) to obtain the LNP preparation obtained by encapsulating Fluc mRNA with the ionizable lipids described in this application and DSPC, cholesterol and DMG-PEG2000. The particle size and polydispersity index (PDI) of each LNP preparation were determined by dynamic light scattering using a Malvern Zetasizer Ultra instrument (manufacturer: Malvern); the encapsulation efficiency of LNP was determined using the Quant-it Ribogreen RNA quantification kit (manufacturer: ThermoFisher Scientific, catalog number: R11490).

[0726] Table 1

[0727]

[0728] In the art, a PDI of less than 0.3 indicates that the nanoparticle size in the LNP preparation is relatively uniform; an encapsulation rate is used to indicate whether the LNP can effectively encapsulate mRNA, and an encapsulation rate of more than 70% indicates that the LNP can effectively encapsulate mRNA. The particle size of the LNP prepared by the present application is maintained at 60 to 100 nm, the PDI is less than 0.15, and the encapsulation rate is higher than 90%.

[0729] Example 2 In vivo animal study of LNP preparations

[0730] In this embodiment, the LNP prepared in Example 1 was injected into female Balb / C mice (Weitong Lihua) aged 6 to 8 weeks at a dose of 5 μg / mouse by tail vein injection or lower limb muscle injection (n=3, i.e., 3 mice were injected and tested in each group, and the data results presented are the mean values ​​of the determinations of each group), and D-luciferin potassium salt was injected intraperitoneally at a specific time point after administration (4 hours, 24 hours, and 48 hours in this embodiment), and then the luminescence was detected by IVIS Spectrum small animal in vivo imager (manufacturer: PerkinElmer), and the total luminescence intensity of the in vivo expression site of the mouse (e.g., liver, lower limb administration site, etc.) was counted. The higher the luminescence intensity, the higher the luciferase expression, i.e., the better the expression of the corresponding LNP preparation in the mouse. The total luminescence intensity is measured by bioluminescence imaging, and the luminescence intensity data of the luminescent site is obtained 6 to 15 minutes (min) after the intraperitoneal injection of D-luciferin potassium salt. The total luminescence intensity of the in vivo expression area was counted by Living Image software (manufacturer: PerkinElmer). Normally, the total luminescence intensity reading of mice not treated with drugs is on the order of 10 5 .

[0731] Referring to the above-mentioned in vivo mouse experimental method, the LNP preparation in Example 1 was injected into 6- to 8-week-old female Balb / C mice via the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the 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.

[0732] Table 2 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 h

[0733] Group Liver AUC (p / s*hour) Compound 1 1.94E+12 Compound 2 2.94E+12

[0734] Example 3 Preparation and Detection of Lipid Nanoparticles (LNP)

[0735] This embodiment selects compound 4, compound 6, compound 7, compound 10, compound 11, compound 12, compound 13, compound 15, compound 20 and compound 21 as ionizable lipids, and prepares LNP preparations (encapsulating Fluc mRNA) in accordance with the molar ratio and nitrogen-phosphorus ratio in Table 3, with reference to Example 1. The mRNA of this embodiment is 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 is used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP preparations in this embodiment are determined. As shown in Table 3, the particle size of the LNP preparations prepared in this embodiment is between 50 and 100 nm, the PDI is less than 0.2, and the encapsulation efficiency is higher than 85%.

[0736] Table 3

[0737]

[0738]

[0739] Example 4 In vivo animal study of LNP preparations

[0740] 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 through the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the mouse liver was counted. The test results are shown in Table 4 and Figure 2 The expression levels of the LNP preparations prepared in this example in mice were: Compound 12>Compound 4>Compound 11>Compound 6>Compound 13>Compound 7>Compound 10>Compound 20>Compound 15>Compound 21.

[0741] Table 4 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[0742] Group Liver AUC (p / s*hour) Compound 4 2.31E+12 Compound 6 2.09E+12 Compound 7 1.32E+12 Compound 10 1.19E+12 Compound 11 2.19E+12 Compound 12 2.63E+12 Compound 13 1.47E+12 Compound 15 1.11E+12 Compound 20 1.19E+12 Compound 21 7.67E+11

[0743] Example 5 Preparation and Detection of Lipid Nanoparticles (LNP)

[0744] This embodiment selects compound 5, compound 8, compound 9, compound 14, compound 22 and compound 23 as ionizable lipids, and prepares LNP preparations (encapsulating FlucmRNA) in accordance with the molar ratio and nitrogen-phosphorus ratio in Table 3, with reference to Example 1. The mRNA of this embodiment 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 embodiment are determined. As shown in Table 5, the particle size of the LNP preparations prepared in this embodiment is between 50 and 80nm, the PDI is less than 0.2, and the encapsulation efficiency is higher than 90%.

[0745] Table 5

[0746]

[0747] Example 6 In vivo animal study of LNP preparations

[0748] 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 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 Table 6 and Figure 3 The expression levels of the LNP preparations prepared in this example in mice were: Compound 22>Compound 23>Compound 14>Compound 9>Compound 5>Compound 8.

[0749] Table 6 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[0750] Group Liver AUC (p / s*hour) Compound 5 1.29E+12 Compound 8 9.21E+11 Compound 9 1.80E+12 Compound 14 1.93E+12 Compound 22 3.25E+12 Compound 23 2.35E+12

[0751] Example 7 Preparation and Detection of Lipid Nanoparticles (LNP)

[0752] This embodiment selects compound 16 and compound 24 as ionizable lipids, and prepares LNP preparations (encapsulating Fluc mRNA) according to the molar ratio and nitrogen-phosphorus ratio in Table 7, with reference to Example 1. The mRNA of this embodiment 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 embodiment are measured. As shown in Table 7, the particle size of the LNP preparations prepared in this embodiment is between 50 and 80nm, the PDI is less than 0.2, and the encapsulation efficiency is higher than 90%.

[0753] Table 7

[0754]

[0755]

[0756] Example 8 In vivo animal studies of LNP preparations

[0757] 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 through the tail vein at a dose of 5 μg / mouse, and the total luminescence intensity of the mouse liver was counted. The test results are shown in Table 8 and Figure 4 , the expression level of the LNP preparation prepared in this example in mice: Compound 16>Compound 24.

[0758] Table 8 Area under the expression kinetic curve (AUC) in the liver region from 4 to 48 hours

[0759] Group Liver AUC (p / s*hour) Compound 16 1.92E+12 Compound 24 8.10E+11 .

Claims

1. A compound I or a pharmaceutically acceptable salt thereof, in, R 1 is a C substituted with one or more hydroxyl groups 1-6 alkyl; X and Y are independently C 1-12 Alkylene; Z 1 and Z 2 Independently W 1 and W 2 Independently a chemical bond or C 1-12 Alkylene; R 2 C 1-15 Alkyl, C 1-15 Alkenyl or -C 1-12 Alkylene-C 3-10 Cycloalkylene-R 21 , where R 21 H or C 1-6 alkyl; R 3 C 1-15 Alkyl, or C 1-15 alkyl; R 4 C 1-15 Alkyl, C 1-15 Alkoxy, C 1-15 Alkenyloxy, -C 1-12 Alkylene-C 3-10 Cycloalkylene-R 41 、-OC 1-12 Alkylene-C 3-10 Cycloalkylene-R 42 ; Among them, R 41 and R 42 are independently H or C 1-6 alkyl; R 5 H, C 1-15 Alkyl, or C 1-15 alkyl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that It meets one or more of the following conditions: (1)R 1 , R 21 , R 41 and R 42 In the above, the "C 1-6 "alkyl" are each independently C 1-4 Alkyl, such as methyl, ethyl, n-propyl or n-butyl; (2)R 1 , R 21 , R 41 and R 42 In the above, the "C 1-6 "Alkyl" is each independently a straight chain or branched C 1-6 alkyl; (3)R 2 and R 4 In the C 3-10 Cycloalkylene is C 3-6 Cycloalkylene, for example, (4)R 1 In the case of a hydroxyl group, only the terminal carbon not connected to the nitrogen atom is replaced by a hydroxyl group. (5) X and Y are independently C 3-7 Alkylene; (6) Z 1 for *Denotes the same as W 1 connect; (7) Z 2 for *Denotes the same as W 2 connect; and (8)W 1 and W 2 Independently chemically bonded or linear C 1-6 Alkylene; Preferably, the compound or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) X and Y are independently C 3-7 Straight chain alkylene; (2)W 1 and W 2 Independently chemically bonded or linear C 1-6 Alkylene; (3)R 2 C 4-9 Alkyl, C 4-9 Alkenyl or -C 4-6 Alkylene-C 3-10 Cycloalkylene-R 21 ; (4)R 3 C 5-10 Alkyl, or a C 5-10 alkyl; (5)R 4 C 8-10 Alkyl, C 5-9 Alkoxy, C 8-10 Alkenyloxy, -C 1-3 Alkylene-C 3-10 Cycloalkylene-R 41 、-OC 4-6 Alkylene-C 3-10 Cycloalkylene-R 42 ; and (6)R 5 H, C 8-10 Alkyl, or a C 8-10 alkyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It meets one or more of the following conditions: (1)R 1 for (2)X is (3)Y is (4)W 1 for (5)W 2 For chemical bonds, (6)R 2 for (7)R 3 for (8)R 4 for and (9)R 5 For H. Preferably, the compound or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) for and (2) for 4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It is Compound IA or a pharmaceutically acceptable salt thereof; 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 3.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: It is any of the following structures:

6. A lipid carrier, characterized in that It includes substance Z, which is compound I or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 5.

7. The lipid carrier according to claim 6, wherein It meets one or more of the following conditions: (1) The lipid carrier further comprises a diluent, which may be a phosphate buffer, a sodium acetate buffer or a Tris-acetate buffer; (2) The lipid carrier further comprises a phospholipid, which may be a phospholipid molecule having an electrically charged polar end and a non-polar fatty chain end, preferably distearoylphosphatidylcholine, dimyristoylphosphocholine, dioleoylphosphocholine, palmitoylphosphocholine, heneicosanoylphosphocholine or palmitoylphosphocholine, such as distearoylphosphatidylcholine; (3) The lipid carrier further comprises PEG lipid, which may be a lipid molecule having a polyethylene glycol hydrophilic end modified; preferably one or more selected from 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; and (4) the lipid carrier further comprises a sterol, which may be an animal, plant or fungal sterol, preferably one or more selected from cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol; Preferably, the lipid carrier satisfies one or more of the following conditions: (1) 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; (2) the molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 2-8:1, for example 3-6:1; (3) The molar ratio of the substance Z to the phospholipid is 1-15:1, preferably 1-10:1, for example 5:1; (4) The molar ratio of substance Z to PEG lipid is 10-100:1, preferably 10-50:1, for example 33.3:1; (5) The molar content of the substance Z is 30 mol% to 70 mol%, for example 50 mol%; (6) The molar content of the phospholipid is 5 mol% to 20 mol%, for example 10 mol%; (7) The molar content of the sterol is 20 mol% to 60 mol%, for example 38.5 mol%; (8) the molar content of the PEG lipid is about 0.2 mol% to 5 mol%, for example 1.5% mol; and (9) the lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.

8. Use of the compound I or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5 or the lipid carrier according to any one of claims 6 to 7 in the preparation of a nucleic acid preventive agent and / or therapeutic agent delivery vector; The nucleic acid preventive and / or therapeutic 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, and is further 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.

9. A lipid nanoparticle, characterized in that: It comprises a nucleic acid preventive and / or therapeutic agent and a lipid carrier as described in any one of claims 6-7; The nucleic acid preventive and / or therapeutic 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, and is further 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.

10. The lipid nanoparticle according to claim 9, characterized in that It meets one or more of the following conditions: (1) The nitrogen-to-phosphorus ratio in the lipid nanoparticles is (2-30):1, preferably, the nitrogen-to-phosphorus ratio is (2-20):1, preferably (3-15):1, for example 6:1; (2) The particle size of the lipid nanoparticle is 10-200 nm, preferably 40-150 nm, and more preferably 50-130 nm, for example, 53.4 nm, 61.9 nm, 66.1 nm, 81.0 nm, 84.0 nm, 78.8 nm, 57.0 nm, 60.6 nm, 61.4 nm, 94.6 nm, 81.7 nm, 58.1 nm, 67.8 nm, 72.1 nm, 57.8 nm, 55.4 nm, 60.7 nm or 64.4 nm; (3) the polydispersity index of the lipid nanoparticles is 0.001-0.3, for example, 0.069, 0.035, 0.030, 0.088, 0.080, 0.075, 0.077, 0.063, 0.055, 0.034, 0.169, 0.112, 0.038, 0.193, 0.178, 0.084, 0.050 or 0.067; (4) The encapsulation efficiency of the lipid nanoparticles is 89%-100%, for example 89-98%; for example 96.1%, 96.6%, 96.8%, 93.0%, 94.7%, 94.0%, 95.4%, 96.3%, 97.4%, 91.3%, 89.7%, 97.6%, 96.9%, 96.2% or 96.4%; and (5) the lipid carrier encapsulates the nucleic acid preventive and / or therapeutic agent.

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