Nitrogen-containing chain compound, preparation method, composition including same, and application

Abstract

Disclosed are a nitrogen-containing chain compound, a preparation method, a composition including same, and an application. Specifically, provided are a nitrogen-containing chain compound as shown in formula (I), or a pharmaceutically acceptable salt thereof. An LNP preparation prepared by using the nitrogen-containing chain compound has a relatively uniform nanoparticle size, a high encapsulation efficiency, and high in vivo expression activity.

Description

Nitrogen-containing chain compounds, methods of making, compositions containing, and uses thereof

[0001] This application claims priority to Chinese Patent Application No. 202410868083X, filed on June 28, 2024, and Chinese Patent Application No. 2024117252319, filed on November 27, 2024. This application incorporates the entirety of the aforementioned Chinese Patent Applications. TECHNICAL FIELD

[0002] The present application relates to a nitrogen-containing chain compound, a method of making, a composition containing, and uses thereof. BACKGROUND

[0003] Nucleic acid drugs are an important direction of current basic and applied research. Nucleic acid drugs can be used for the prevention and / or treatment of viral and bacterial infectious diseases, tumors, metabolic diseases, etc., and have lower generation costs and shorter cycles, which are beneficial for the rapid development of personalized drugs. However, nucleic acids are large molecules with negative charges, which are difficult to penetrate the cell membrane. At the same time, nucleic acids have poor stability. The development of various nucleic acid packaging and delivery systems can overcome the instability of nucleic acid drugs to some extent and improve their delivery efficiency.

[0004] Lipid nanoparticles have been shown to be useful as carriers for delivering biologically active substances, such as small molecule drugs, proteins, and nucleic acids, into cells and / or intracellular compartments. By designing and optimizing the types and amounts of components in the lipid nanoparticles, it is more important to optimize the nucleic acid drug delivery system for improving the efficacy of nucleic acid drug prevention and treatment, especially lipid compounds that can be used to deliver RNA prophylactic and / or therapeutic agents, and related methods and compositions. SUMMARY

[0005] In order to overcome the problems of the prior art that the types of ionizable lipid compounds and nucleic acid prophylactic and / or therapeutic agent delivery carriers are few and the effect is poor, the present application provides a nitrogen-containing chain compound, a method of making, a composition containing, and uses thereof. The composition of the present application can be used for efficient delivery of nucleic acid drugs. The LNP preparation prepared by using the nitrogen-containing chain compound of the present application 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 application solves the above technical problems by the following technical solutions.

[0007] The present application provides a compound I or a pharmaceutically acceptable salt thereof:

[0008] wherein R 1 is C 1-6 alkyl substituted with one or more hydroxyl groups;

[0009] X and Y are independently C 3-12 alkylene;

[0010] Z 1 and Z 2 are independently

[0011] W 1 is a bond, * indicates the attachment to Z 1 ;

[0012] W 2 is * indicates the attachment to Z 2 ;

[0013] R w11 , R w12 , R w21 and R w22 are independently C 1-5 alkylene;

[0014] R 2 is C 1-24 alkyl;

[0015] R 3 is C 1-24 alkyl.

[0016] In certain preferred embodiments of the present application, certain groups of the compounds I or their pharmaceutically acceptable salts are defined as follows, the groups not mentioned are as described in any of the embodiments of the present application (simply "in an embodiment of the present application").

[0017] In an embodiment of the present application, R 1 is C 1-6 alkyl substituted by one hydroxy group.

[0018] In an embodiment of the present application, in R 1 only the terminal carbon of the end not attached to the N atom is substituted by a hydroxy group.

[0019] In an embodiment of the present application, R 1 is

[0020] In an embodiment of the present application, X and Y are independently C 3-7 alkylene, preferably C 3-6 alkylene.

[0021] In an embodiment of the present application, X and Y are independently straight chain alkylene.

[0022] In an embodiment of the present application, X and Y are independently

[0023] In one embodiment of the application, Z 1 is * denotes the attachment to W 1 .

[0024] In one embodiment of the application, Z 2 is * denotes the attachment to W 2 .

[0025] In one embodiment of the application, R w11 , R w12 , R w21 and R w22 are each independently a C 2-4 straight-chain alkylene group.

[0026] In one embodiment of the application, W 1 is a chemical bond, * denotes the attachment to Z 1 .

[0027] In one embodiment of the application, W 2 is * denotes the attachment to Z 2 .

[0028] In one embodiment of the application, R 2 is a straight-chain or branched C 4-18 alkyl group; preferably a straight-chain C 5-11 alkyl group or a branched C 15-17 alkyl group.

[0029] In one embodiment of the application, R 2 is

[0030] In one embodiment of the application, R 3 is R 31 and R 32 are each independently a C 3-10 alkyl group, for example a C 6-9 straight-chain alkyl group.

[0031] In one embodiment of the application, R 31 and R 32 are each independently

[0032] In one embodiment of the application, R 3 is

[0033] In one embodiment of the present application, For

[0034] In one embodiment of the present application, For

[0035] In one embodiment of the present application, the compound I is a structure as shown in I-A, I-B or I-C:

[0036] wherein R 1 , X, Y, Z 1 , W 1 , R 2 , R w21 , R w22 and R 3 are as defined in any of the preceding embodiments.

[0037] In one embodiment of the present application, the compound I is any of the following structures:

[0038] The present application also provides a use of the compound I or a pharmaceutically acceptable salt thereof in the preparation of a nucleic acid prophylactic and / or therapeutic delivery vehicle.

[0039] The nucleic acid therapeutic and / or prophylactic is preferably one or more of a 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), more preferably mRNA, such as firefly luciferase (Fluc) mRNA, SARS-CoV-2 Spike mRNA, varicella-zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0040] The present application also provides a composition comprising a substance Z, which is a compound I or a pharmaceutically acceptable salt thereof as described previously.

[0041] The present application also provides a lipid carrier comprising a substance Z, wherein the substance Z is a compound I or a pharmaceutically acceptable salt thereof as described above.

[0042] In an embodiment of the present application, the lipid carrier further comprises a diluent. The diluent can be a phosphate buffer, a sodium acetate buffer or a Tris-acetate buffer, etc.

[0043] In an embodiment of the present application, the lipid carrier further comprises a phospholipid.

[0044] In an embodiment of the present application, the phospholipid can be a conventional phospholipid in the art, which is an amphipathic auxiliary molecule, and is helpful for the fusion of lipid particles and cell membranes. The phospholipid can be a phospholipid molecule having a charged polar end and a non-polar end of a fatty chain, such as distearoylphosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), palmitoylphosphatidylcholine (DPPC), heneicosanoylphosphatidylcholine (DUPC), or palmitoylphosphatidylcholine (POPC), etc., preferably distearoylphosphatidylcholine.

[0045] In an embodiment of the present application, the lipid carrier further comprises a PEG lipid (polyethylene glycol modified lipid).

[0046] In an embodiment of the present application, the PEG lipid can be a lipid molecule modified with a hydrophilic end of polyethylene glycol. 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), etc.

[0047] In an embodiment of the present application, the lipid carrier further comprises a sterol.

[0048] In an embodiment of the present application, the sterol can be a conventional sterol in the art, and the sterol includes animal, plant or fungal sterol. The sterol is selected from one or more of cholesterols, sitosterols, ergosterols, campesterols, stigmasterols, brassicasterols, tomatine, ursolic acid and α-tocopherol, such as cholesterols, etc.

[0049] In an embodiment of the present application, in the lipid carrier, the molar ratio of the substance Z to the sterol is (0.5-5):1, preferably (0.5-3):1, such as 1.3:1.

[0050] In an embodiment of the present application, 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.

[0051] In one embodiment of the present application, the molar ratio of the substance Z to the phospholipid in the lipid carrier is (1-15): 1, preferably (1-10): 1, for example 5: 1.

[0052] In one embodiment of the present application, the molar ratio of the substance Z to the PEG lipid in the lipid carrier is (10-100): 1, preferably (10-50): 1, for example 33.3: 1.

[0053] In the present application, 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 is not more than 100 mol%.

[0054] In one embodiment of the present application, the molar content of the substance Z is 30 mol% to 70 mol%, for example 50 mol%.

[0055] In one embodiment of the present application, the molar content of the phospholipid is 5 mol% to 20 mol%, for example 10 mol%.

[0056] In one embodiment of the present application, the molar content of the sterol is 20 mol% to 60 mol%, for example 38.5 mol%.

[0057] In one embodiment of the present application, the molar content of the PEG lipid is about 0.2 mol% to 5 mol%, for example 1.5% mol.

[0058] In one embodiment of the present application, the lipid carrier consists of the substance Z, the diluent, the phospholipid, the PEG lipid and the sterol.

[0059] The present application also provides a use of the lipid carrier in the preparation of a nucleic acid prophylactic and / or therapeutic delivery carrier.

[0060] The nucleic acid therapeutic and / or prophylactic 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, for example firefly luciferase (Fluc) mRNA, SARS-CoV-2 Spike mRNA, varicella-zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA or influenza virus (Flu) mRNA.

[0061] The present application also provides a lipid nanoparticle comprising a nucleic acid prophylactic and / or therapeutic and the aforementioned lipid carrier.

[0062] The nucleic acid therapeutic and / or prophylactic agent is preferably one or more of a 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 mRNA, varicella-zoster virus (VZV) mRNA, respiratory syncytial virus (RSV) mRNA, or influenza virus (Flu) mRNA.

[0063] In an embodiment of the present application, the nitrogen to phosphorus ratio in the lipid nanoparticle is 2-30:1; preferably 3-16:1; such as 6:1; the nitrogen to phosphorus ratio refers to the ratio of the moles of ionizable nitrogen atoms in the substance Z to the moles of phosphate groups in the RNA in the lipid nanoparticle.

[0064] In an embodiment of the present application, the lipid nanoparticle has a particle size (average particle size) of 10-200 nm, preferably 40-150 nm, such as 50.32 nm, 53.71 nm, 59.16 nm, 61.74 nm, 67.40 nm, 84.31 nm, 63.89 nm, 95.80, 100.27, 73.07, 62.60, 97.72, 131.87, or 131.40; such as 84.31 nm, 63.89 nm, 95.80, 100.27, 73.07, 62.60, or 131.40.

[0065] In an embodiment of the present application, the lipid nanoparticle has a polydispersity index of 0.001-0.3; such as 0.135, 0.323, 0.107, 0.089, 0.106, 0.188, 0.201, 0.129, 0.079, 0.159, 0.298, 0.235, or 0.073; such as 0.129, 0.079, 0.159, 0.298, 0.235, or 0.073.

[0066] In an embodiment of the present application, the lipid nanoparticle has an encapsulation efficiency of 84%-100%, such as 84.5%, 92.5%, 93.4%, 96.3%, 94.9%, 94.1%, 92.9%, 96.4%, 96.7%, 92.7%, 93.5%, 96.2%, 96.1%, or 86.8%.

[0067] In some embodiments of the present application, the encapsulation efficiency of the lipid nanoparticle is 85-100%, such as 96.40% or 96.66%, 92.74%, 93.51%, 96.17%, 96.10% or 86.82%.

[0068] In some embodiments of the present application, the lipid carrier encapsulates the nucleic acid prophylactic and / or therapeutic agent in the lipid nanoparticle.

[0069] On the basis of common general knowledge in the art, the above-mentioned preferred conditions can be combined in any manner, thereby obtaining various preferred embodiments of the present application.

[0070] The reagents and raw materials used in the present application are commercially available.

[0071] The positive progress effect of the present application is that the LNP preparation prepared by using the nitrogen-containing chain compound of the present application has relatively uniform nanoparticle size, high encapsulation efficiency, and high in vivo expression activity. BRIEF DESCRIPTION OF DRAWINGS

[0072] Figure 1 is a plot of the chemiluminescence intensity in the liver region of mice over time for Compound 2 and Compound 3 administered intravenously;

[0073] Figure 2 is a plot of the chemiluminescence intensity in the administration site of mice over time for Compound 2 and Compound 3 administered intramuscularly;

[0074] Figure 3 is a plot of the chemiluminescence intensity in the liver region of mice over time for Compound 2 and Compound 3 administered intramuscularly;

[0075] Figure 4 is a plot of the chemiluminescence intensity in the liver region of mice over time for Compound 4 and Compound 5 administered intravenously;

[0076] Figure 5 is a plot of the chemiluminescence intensity in the liver region of mice over time for Compound 4 and Compound 5 administered intravenously;

[0077] Figure 6 is a plot of the chemiluminescence intensity in the liver region of mice over time for Compound 1 administered intravenously.

[0078] Figure 7 is a plot of the chemiluminescence intensity in the liver region of mice over time for H21, H23, Compound 10, Compound 11, Compound 12, Compound 14 and Compound 17 administered intravenously.

[0079] Figure 8 is a plot of the chemiluminescence intensity in the liver region of mice over time for Compound 6 and Compound 8 administered intravenously. DETAILED DESCRIPTION

[0080] The present application is further illustrated by the following examples without limiting the present application to the examples.

[0081] Preparation of compound 2

[0082] Step 1: Preparation of 2-1

[0083] Reaction formula:

[0084] Material ratio:

[0085] Operation process:

[0086] Into a reaction bottle was added succinic anhydride, 8-pentadecanol, DMAP, triethylamine and DCM, and the mixture was stirred at room temperature for 16 h. The product had an Rf value of 0.6, TLC (DCM:MeOH=10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After column chromatography, 6.5 g of colorless oil was obtained.

[0087] Step 2: Preparation of 2-2

[0088] Reaction formula:

[0089] Material ratio:

[0090] Operation process:

[0091] Into a reaction bottle was added 2-1, 6-bromohexanol, EDCI, DMAP and DCM, and the mixture was stirred at room temperature for 16 h. The product had an Rf value of 0.5, TLC (PE:EA=10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After column chromatography, 5.6 g of colorless oil was obtained.

[0092] Step 3: Preparation of compound 2

[0093] Reaction formula:

[0094] Material ratio:

[0095] Operation process:

[0096] Into a reaction bottle was added 2-2, SM102-3, potassium carbonate, potassium iodide and acetonitrile, and the mixture was stirred at 65°C for 16 h. The product had an Rf value of 0.5, TLC (DCM:MeOH=10:1). The reaction solution was concentrated, diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After column chromatography, 1.6 g of colorless oil was obtained.

[0097] 1H NMR (600 MHz, Chloroform-d) δ 5.08 (qd, J = 7.0, 3.2 Hz, 1H), 4.22 (dd, J = 11.8, 3.3 Hz, 1H), 4.07 - 3.99 (m, 3H), 3.53 (t, J = 5.3 Hz, 2H), 2.57 (t, J = 5.4 Hz, 2H), 2.46 (q, J = 8.3 Hz, 4H), 2.33 - 2.27 (m, 6H), 1.65 - 1.53 (m, 10H), 1.50 - 1.43 (m, 4H), 1.34 - 1.22 (m, 38H), 0.87 (t, J = 6.8 Hz, 9H).

[0098] MS (ES+) m / z): 740.6 (M+H) + .

[0099] Preparation of compound 3

[0100] Step 1: Preparation of compound 3

[0101] Reaction formula:

[0102] Material ratio:

[0103] Operation process:

[0104] Into a reaction bottle was added 2-2, ethanolamine, potassium carbonate, potassium iodide and acetonitrile, and stirred at 65°C for 16h. The product had an Rf value of 0.5, TLC (DCM:MeOH=10:1). The reaction solution was concentrated, then diluted with 100 mL of ethyl acetate, washed with 100 mL of water once, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and then purified by column chromatography to obtain 1.6g of colorless oil.

[0105] 1 H NMR (600 MHz, Chloroform-d) δ 4.89 - 4.84 (m, 2H), 4.11 (q, J = 7.1 Hz, 1H), 4.07 (t, J = 6.7 Hz, 4H), 3.55 (t, J = 5.3 Hz, 2H), 2.61 (s, 10H), 2.48 (t, J = 7.6 Hz, 4H), 1.62 (p, J = 6.9 Hz, 4H), 1.54 - 1.44 (m, 12H), 1.37 - 1.22 (m, 48H), 0.87 (t, J = 7.0 Hz, 12H).

[0106] MS (ES+) m / z): 882.7 (M+H) + .

[0107] Preparation of compound 4

[0108] Step one: Synthesis of 4-(heptadecan-9-yloxy)-4-oxo butyric acid

[0109] Reaction scheme:

[0110] Material ratio:

[0111] Operation process:

[0112] Into a reaction flask was added tetrahydrofuran (30.0 mL), heptadecan-9-ol (3.00 g, 11.7 mmol), succinic anhydride (1.76 g, 17.5 mmol), 4-dimethylaminopyridine (1.43 g, 11.7 mmol), pyridine (1.39 g, 17.5 mmol) successively, and the mixture was reacted at 60 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC, and the reaction solution was concentrated. Purification by column chromatography gave 4-(heptadecan-9-yloxy)-4-oxo butyric acid (3.74 g) as a colorless liquid.

[0113] Step two: Synthesis of 1-(6-bromohexyl) 4-heptadecan-9-yl butyrate

[0114] Reaction scheme:

[0115] Material ratio:

[0116] Operation process:

[0117] Into a reaction flask was added tetrahydrofuran (30.0 mL), heptadecan-9-ol (3.00 g, 11.7 mmol), succinic anhydride (1.76 g, 17.5 mmol), 4-dimethylaminopyridine (1.43 g, 11.7 mmol), pyridine (1.39 g, 17.5 mmol) successively, and the mixture was reacted at 60 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC, and the reaction solution was concentrated. Purification by column chromatography gave 4-(heptadecan-9-yloxy)-4-oxo butyric acid (3.74 g) as a colorless liquid.

[0118] Step three: Synthesis of 1-heptadecan-9-yl 4-{6-[(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino]hexyl} butyrate

[0119] Reaction scheme:

[0120] Material ratio:

[0121] Procedure:

[0122] Dissolve undecyl 6-[(2-hydroxyethyl)amino]hexanoate (600 mg, 1.82 mmol) in acetonitrile (6.00 mL), add 1-(6-bromohexyl) 4-heptadecan-9-yl butanedioate (1.23 g, 2.37 mmol), potassium carbonate (880 mg, 6.37 mmol), potassium iodide (362 mg, 2.19 mmol), tetrahydrofuran (6.00 mL) in sequence, and finally react at 80 °C for 12 hours under nitrogen protection. Monitor the reaction by TLC until completion, filter, and concentrate. Purify by column chromatography to obtain 1-heptadecan-9-yl 4-{6-[(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino]hexyl} butanedioate (715 mg) as a colorless liquid.

[0123] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.88 (t, J = 6.2 Hz, 1H), 4.07 (q, J = 6.8 Hz, 4H), 3.53 (t, J = 5.2 Hz, 2H), 2.62 (s, 4H), 2.58 (t, J = 5.2 Hz, 2H), 2.49 - 2.41 (m, 4H), 2.31 (t, J = 7.4 Hz, 2H), 1.63 - 1.42 (m, 14H), 1.39 - 1.18 (m, 46H), 0.99 - 0.83 (m, 9H) ppm.

[0124] LCMS: RT = 2.015, m / z 767 [M+H] +

[0125] Preparation of compound 5

[0126] Step one: synthesis of 6-(heptadecan-9-yloxy)-6-oxohexanoic acid

[0127] Reaction scheme:

[0128] Material ratio:

[0129] Procedure:

[0130] In a reaction flask, 6-(heptadecan-9-yloxy)-6-oxohexanoic acid (1.90 g, 4.94 mmol) was dissolved in tetrahydrofuran (20 mL), followed by the addition of 6-bromohexan-1-ol (1.79 g, 9.88 mmol), 4-dimethylaminopyridine (120 mg, 988 μmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.42 g, 7.41 mmol), and finally 40 °C for 16 hours. TLC monitoring of the reaction was completed, and the reaction solution was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by column chromatography to obtain 1-(6-bromohexyl) 6-heptadecan-9-yl hexanedioate (2.30 g) as a colorless liquid.

[0131] Step Two: Synthesis of 1-(6-bromohexyl) 6-heptadecan-9-yl hexanedioate

[0132] Reaction Formula:

[0133] Material Ratio:

[0134] Operation Process:

[0135] In a reaction flask, 6-(heptadecan-9-yloxy)-6-oxohexanoic acid (1.90 g, 4.94 mmol) was dissolved in tetrahydrofuran (20 mL), followed by the addition of 6-bromohexan-1-ol (1.79 g, 9.88 mmol), 4-dimethylaminopyridine (120 mg, 988 μmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (1.42 g, 7.41 mmol), and finally 40 °C for 16 hours. TLC monitoring of the reaction was completed, and the reaction solution was extracted with ethyl acetate three times, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was purified by column chromatography to obtain 1-(6-bromohexyl) 6-heptadecan-9-yl hexanedioate (2.30 g) as a colorless liquid.

[0136] Step Three: Synthesis of 1-heptadecan-9-yl 6-{6-[(2-hydroxyethyl)[6-oxo-6- (undecyloxy)hexyl]amino]hexyl} hexanedioate

[0137] Reaction Formula:

[0138] Material Ratio:

[0139] Operation Process:

[0140] In a reaction flask, 1-(6-bromohexyl) 6-heptadecan-9-yl hexanedioate (1.00 g, 1.83 mmol) was dissolved in acetonitrile (10 mL), then added 1-heptadecan-9-yl 6-[(2-hydroxyethyl)amino]hexanoate (661 mg, 2.01 mmol), potassium carbonate (757 mg, 5.48 mmol), potassium iodide (363 mg, 2.19 mmol), tetrahydrofuran (10 mL) successively, and finally reacted at 80 °C for 12 hours under nitrogen protection. The reaction was monitored by TLC, and the reaction was completed. The crude product was concentrated, and the crude product was purified by column chromatography to obtain 1-heptadecan-9-yl 6-{6-[(2-hydroxyethyl)[6-oxo-6-(heptadecyloxy)hexyl]amino]hexyl} hexanedioate (1.10 g) as a colorless liquid.

[0141] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.86 (quin, J = 6.2 Hz, 1H), 4.06 (t, J = 6.6 Hz, 4H), 3.74 (br s, 2H), 2.83 (br s, 2H), 2.72 (br s, 4H), 2.36-2.29 (m, 6H), 1.71-1.58 (m, 14H), 1.51 (br d, J = 5.4 Hz, 4H), 1.41-1.23 (m, 46H), 0.88 (br t, J = 6.6 Hz, 9H).

[0142] LCMS: RT = 2.054, m / z 796.7 [M+H] +

[0143] Preparation of compound 7

[0144] Step one: synthesis of 1-(4-bromobutyl) 6-heptadecan-9-yl hexanedioate

[0145] Reaction formula:

[0146] Material ratio:

[0147] Operation process:

[0148] In a reaction flask, 6-(heptadecan-9-yloxy)-6-oxohexanoic acid (1.10 g, 2.86 mmol) was dissolved in tetrahydrofuran (12 mL), 4-bromobutane-1-ol (875 mg, 5.72 mmol) was added, followed by 4-dimethylaminopyridine (69.8 mg, 572 μmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (822 mg, 4.29 mmol), and finally 40 °C for 16 h. TLC monitoring of the reaction was completed by extraction with ethyl acetate three times, dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by column chromatography to give 1-(4-bromobutyl) 6-heptadecan-9-yl hexanedioate (800 mg) as a yellow liquid.

[0149] Step two: synthesis of 1-heptadecan-9-yl 6-{4-[(2-hydroxyethyl)[6-oxo-6- (undecyloxy)hexyl]amino]butyl} hexanedioate

[0150] Material ratio:

[0151] Operation process:

[0152] In a reaction flask, 1-(4-bromobutyl) 6-heptadecan-9-yl hexanedioate (800 mg, 1.54 mmol) was dissolved in acetonitrile (5 mL), followed by the addition of undecyl 6-[(2-hydroxyethyl)amino]hexanoate (558 mg, 1.69 mmol), potassium carbonate (638 mg, 4.62 mmol), potassium iodide (306 mg, 1.85 mmol), tetrahydrofuran (5 mL), and finally 80 °C for 12 h under nitrogen protection. TLC monitoring of the reaction was completed by extraction with ethyl acetate three times, dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was purified by column chromatography to give 1-heptadecan-9-yl 6-{4-[(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino]butyl} hexanedioate (500 mg) as a yellow liquid.

[0153] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.87 (quin, J = 6.4 Hz, 1H), 4.08 (td, J1 = 6.6, J2 = 10.2 Hz, 4H), 3.62 (br s, 2H), 2.73-2.53 (m, 6H), 2.36-2.29 (m, 6H), 1.69-1.48 (m, 20H), 1.26 (br s, 40H), 0.89 (t, J = 6.8 Hz, 9H).

[0154] LCMS: RT = 2.033, m / z 768.6 [M+H] +

[0155] Preparation of compound 9

[0156] Step 1: Preparation of 9-1

[0157] Reaction formula:

[0158] Material ratio:

[0159] Operation process:

[0160] Into a reaction bottle was added succinic anhydride, 8-heptadecanol, DMAP, triethylamine and DCM, and stirred at room temperature for 16 h. The product had an Rf value of 0.6, TLC (DCM:MeOH=10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After column chromatography, 6 g of colorless oil was obtained.

[0161] Step 2: Preparation of 9-2

[0162] Reaction formula:

[0163] Material ratio:

[0164] Operation process:

[0165] Into a reaction bottle was added 9-1, 6-bromohexanol, EDCI, DMAP and DCM, and stirred at room temperature for 16 h. The product had an Rf value of 0.5, TLC (PE:EA=10:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After column chromatography, 5.6 g of colorless oil was obtained.

[0166] Step 3: Preparation of compound 9

[0167] Reaction formula:

[0168] Material ratio:

[0169] Operation process:

[0170] Into a reaction bottle was added 9-2, ethanolamine, potassium carbonate, potassium iodide and acetonitrile, and stirred at 65°C for 16 h. The product had an Rf value of 0.5, TLC (DCM:MeOH=10:1). The reaction solution was concentrated, then diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. After column chromatography, 2 g of colorless oil was obtained.

[0171] 1H NMR (600 MHz, Chloroform-d) δ 4.89 - 4.84 (m, 2H), 4.11 (q, J = 7.1 Hz, 1H), 4.07 (t, J = 6.7 Hz, 4H), 3.55 (t, J = 5.3 Hz, 2H), 2.61 (s, 10H), 2.48 (t, J = 7.6 Hz, 4H), 1.62 (p, J = 6.9 Hz, 4H), 1.54 - 1.44 (m, 12H), 1.37 - 1.22 (m, 56H), 0.87 (t, J = 7.0 Hz, 12H).

[0172] MS (ES+) m / z): 938.7 (M+H) + .

[0173] Preparation of compound 1

[0174] Step 1: Preparation of 1-1

[0175] Reaction formula:

[0176] Material ratio:

[0177] Operation process:

[0178] Into the reaction bottle, 1,4-butanediol, n-hexanoic acid, EDCI, DMAP and DCM were added, and the reaction was stirred at room temperature for 16 h. The product had an Rf value of 0.4, TLC (PE:EA=10:1). The reaction solution was washed once with 200 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After concentration, 12 g of oil was obtained after column chromatography purification.

[0179] Step 2: Preparation of 1-2

[0180] Reaction formula:

[0181] Material ratio:

[0182] Operation process:

[0183] Into the reaction bottle, 1-1, 6-bromohexanoic acid, EDCI, DMAP and DCM were added, and the reaction was stirred at room temperature for 16 h. The product had an Rf value of 0.5, TLC (PE:EA=20:1). The reaction solution was washed once with 150 mL of water, and the organic phase was dried over anhydrous sodium sulfate. After concentration, 6 g of oil was obtained after column chromatography purification.

[0184] Step 3: Preparation of 1-3

[0185] Reaction formula:

[0186] Material ratio:

[0187] Procedure:

[0188] Into a flask was added succinic anhydride, 7-tridecanol, DMAP, triethylamine and DCM, and stirred at room temperature for 16 h. The reaction solution was concentrated, and 6 g of colorless oil was obtained after purification by column chromatography. The product had an Rf value of 0.6, TLC (DCM:MeOH=10:1). The reaction solution was concentrated, and 6 g of white solid was obtained after purification by column chromatography.

[0189] Step 4: Preparation of compound 1-4

[0190] Reaction scheme:

[0191] Material ratio:

[0192] Procedure:

[0193] Into a flask was added 1-3, 6-bromohexanol, EDCI, DMAP and DCM, and stirred at room temperature for 16 h. The product had an Rf value of 0.6, TLC (PE:EA=20:1). The reaction solution was washed once with 100 mL of water, and the organic phase was dried with anhydrous sodium sulfate, concentrated, and 4 g of oil was obtained after purification by column chromatography.

[0194] Step 5: Preparation of 1-5

[0195] Reaction scheme:

[0196] Material ratio:

[0197] Procedure:

[0198] Into a flask was added 1-4, ethanolamine and acetonitrile, and stirred at room temperature for 16 h. The product had an Rf value of 0.2, TLC (DCM:MeOH=10:1). The reaction solution was concentrated, diluted with 100 mL of ethyl acetate, washed twice with 100 mL of water, and the organic phase was dried with anhydrous sodium sulfate, concentrated, and 2 g of oil was obtained after purification by column chromatography.

[0199] Step 6: Preparation of compound 1

[0200] Reaction scheme:

[0201] Material ratio:

[0202] Procedure:

[0203] Into a reaction flask was added 1-2, 1-5, potassium carbonate, potassium iodide and acetonitrile, and stirred at 65 °C for 16 h. The product had an Rf value of 0.6, TLC (DCM:MeOH=10:1). The reaction mixture was concentrated and diluted with 100 mL of ethyl acetate, washed with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to give 1.5 g of an oil.

[0204] 1 H NMR (600 MHz, Chloroform-d) δ 4.84 (h, J = 5.7 Hz, 1H), 4.09 - 4.02 (m, 6H), 3.58 (t, J = 5.3 Hz, 2H), 2.65 (t, J = 5.3 Hz, 2H), 2.59 (d, J = 2.2 Hz, 4H), 2.56 - 2.50 (m, 4H), 2.28 (dt, J = 10.2, 7.5 Hz, 4H), 1.67 (h, J = 3.0 Hz, 4H), 1.64 - 1.57 (m, 6H), 1.49 (td, J = 10.9, 7.9, 3.4 Hz, 8H), 1.35 - 1.19 (m, 26H), 0.86 (dt, J = 12.5, 6.9 Hz, 9H).

[0205] MS (ES+) m / z): 728.6 (M+H) + .

[0206] Preparation of compound 10

[0207] Step 1: Preparation of 10-1

[0208] Reaction scheme:

[0209] Material ratio:

[0210] Operation process:

[0211] Into a reaction flask was added succinic anhydride, 8-pentadecanol, DMAP, triethylamine and DCM, and stirred at room temperature for 16 h. The product had an Rf value of 0.6, TLC (DCM:MeOH=10:1). The reaction mixture was concentrated and purified by column chromatography to give 6 g of a colorless oil.

[0212] Step 2: Preparation of 10-2

[0213] Reaction scheme:

[0214] Material ratio:

[0215] Operation process:

[0216] To a flask was added 10-1, 3-bromopropanol, EDCI, DMAP and DCM and stirred at room temperature for 2 h. The product had an Rf value of 0.5, TLC (PE:EA = 10:1). The reaction was washed once with 80 mL of water and the organic phase was dried over anhydrous sodium sulfate, concentrated and purified by column chromatography to give 3.8 g of an oil.

[0217] Step 3: Preparation of compound 10

[0218] Reaction Scheme:

[0219] Material Ratio:

[0220] Operation Process:

[0221] To a flask was added 10-2, ethanolamine, potassium carbonate, potassium iodide and acetonitrile and heated to 65 °C and stirred for 16 h. The product had an Rf value of 0.5, TLC (DCM:MeOH = 10:1). The reaction was diluted with 100 mL of ethyl acetate and washed once with 100 mL of water and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated and purified by column chromatography to give 2 g of a colorless oil.

[0222] 1 H NMR (400 MHz, Chloroform-d) δ 4.86 (p, J = 6.2 Hz, 2H), 4.13 (t, J = 6.3 Hz, 4H), 3.55 (t, J = 5.3 Hz, 2H), 2.58 (d, J = 25.4 Hz, 14H), 1.78 (p, J = 6.6 Hz, 4H), 1.50 (q, J = 6.2 Hz, 8H), 1.26 (d, J = 6.1 Hz, 40H), 0.87 (t, J = 6.6 Hz, 12H).

[0223] MS (ES+) m / z): 798.6 (M) + .

[0224] Preparation of compound 11

[0225] Step 1: Preparation of 11-1

[0226] Reaction Scheme:

[0227] Material Ratio:

[0228] Operation Process:

[0229] Into a reaction flask was placed 1,6-hexanedioic acid, 8-pentadecanol, EDCI, DMAP and DCM, and the reaction was stirred at room temperature for 2 h. The product had an Rf value of 0.6, TLC (DCM:MeOH = 10:1). The reaction was concentrated and purified by column chromatography to give 6 g of colorless oil.

[0230] Step 2: Preparation of 11-2

[0231] Reaction Scheme:

[0232] Material Ratio:

[0233] Operation Process:

[0234] Into a reaction flask was placed 11-1, 4-bromobutanol, EDCI, DMAP and DCM, and the reaction was stirred at room temperature for 2 h. The product had an Rf value of 0.5, TLC (PE:EA = 10:1). The reaction was washed once with 80 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to give 3.8 g of oil.

[0235] Step 3: Preparation of Compound 11

[0236] Reaction Scheme:

[0237] Material Ratio:

[0238] Operation Process:

[0239] Into a reaction flask was placed 11-2, ethanolamine, potassium carbonate, potassium iodide and acetonitrile, and the reaction was stirred at 65 °C for 16 h. The product had an Rf value of 0.5, TLC (DCM:MeOH = 10:1). The reaction was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to give 2 g of colorless oil.

[0240] 1 H NMR (400 MHz, Chloroform-d) δ 4.86 (p, J = 6.3 Hz, 2H), 4.07 (t, J = 6.6 Hz, 4H), 3.54 (t, J = 5.4 Hz, 2H), 2.58 (t, J = 5.3 Hz, 2H), 2.48 (t, J = 7.3 Hz, 4H), 2.36 - 2.27 (m, 8H), 1.67 - 1.60 (m, 12H), 1.50 (d, J = 6.9 Hz, 12H), 1.25 (s, 40H), 0.87 (t, J = 6.7 Hz, 12H).

[0241] MS (ES+) m / z): 883.0 (M+H)+ .

[0242] Preparation of compound 12

[0243] Step 1: Preparation of 12-1

[0244] Reaction Scheme:

[0245] Material ratio:

[0246] Operation process:

[0247] Into a reaction flask was added 11-1, 6-bromohexanol, EDCI, DMAP and DCM, and the mixture was stirred at room temperature for 2 h. The product had an Rf value of 0.5 by TLC (PE:EA = 10:1). The reaction solution was washed once with 80 mL of water, and the organic phase was dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 3.8 g of an oil.

[0248] Step 2: Preparation of compound 12

[0249] Reaction Scheme:

[0250] Material ratio:

[0251] Operation process:

[0252] Into a reaction flask was added 12-1, ethanolamine, potassium carbonate, potassium iodide and acetonitrile, and the mixture was stirred at 65°C for 16 h. The product had an Rf value of 0.5 by TLC (DCM:MeOH = 10:1). The reaction solution was diluted with 100 mL of ethyl acetate, washed once with 100 mL of water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain 2 g of a colorless oil.

[0253] 1 H NMR (400 MHz, Chloroform-d) δ 4.86 (p, J = 6.2 Hz, 2H), 4.05 (t, J = 6.7 Hz, 4H), 3.56 (t, J = 5.3 Hz, 2H), 2.62 (t, J = 5.3 Hz, 2H), 2.49 (t, J = 7.5 Hz, 4H), 2.31 (q, J = 6.5 Hz, 8H), 1.68 - 1.61 (m, 12H), 1.47 (dd, J = 16.4, 9.3 Hz, 12H), 1.35 - 1.22 (m, 48H), 0.87 (t, J = 6.7 Hz, 12H).

[0254] MS (ES+) m / z): 938.7 (M) + .

[0255] Preparation of compound 14

[0256] Step one: synthesis of benzyl 3-(l-heptyloctoxycarbonyloxy)propylate

[0257] Reaction scheme:

[0258] Material ratio:

[0259] Operation process:

[0260] Benzyl 3-(l-heptyloctoxycarbonyloxy)propylate (4.60 g, 10.58 mmol) was dissolved in methanol (10.0 mL), palladium on carbon (1.13 g, 1.06 mmol) was added under argon atmosphere, finally the reaction was carried out under hydrogen (40 Psi) atmosphere at 35 °C for 12 hours, TLC monitored the end of the reaction, the reaction was filtered and concentrated. The crude product was dried under vacuum and purified by column chromatography to give 3-(l-heptyloctoxycarbonyloxy)propanoic acid (2.50 g) as colorless liquid.

[0261] Step two: synthesis of 3-(l-heptyloctoxycarbonyloxy)propanoic acid

[0262] Reaction scheme:

[0263] Material ratio:

[0264] Operation process:

[0265] Benzyl 3-(l-heptyloctoxycarbonyloxy)propylate (4.60 g, 10.58 mmol) was dissolved in methanol (10.0 mL), palladium on carbon (1.13 g, 1.06 mmol) was added under argon atmosphere, finally the reaction was carried out under hydrogen (40 Psi) atmosphere at 35 °C for 12 hours, TLC monitored the end of the reaction, the reaction was filtered and concentrated. The crude product was dried under vacuum and purified by column chromatography to give 3-(l-heptyloctoxycarbonyloxy)propanoic acid (2.50 g) as colorless liquid.

[0266] Step three: synthesis of 6-bromohexyl 3-(l-heptyloctoxycarbonyloxy)propylate

[0267] Reaction scheme:

[0268] Material ratio:

[0269] Procedure:

[0270] Dissolve 3-(1-heptyloctoxycarbonyloxy)propanoic acid (1.00 g, 2.90 mmol) in dichloromethane (10.0 mL), add 6-bromohexan-1-ol (1.42 g, 3.77 mmol), 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.83 g, 4.35 mmol), 4- dimethylaminopyridine (35.5 mg, 290 μmol) successively, and then react at 25 °C for 12 hours under nitrogen protection. TLC is used to monitor the end of the reaction, and the reaction solution is concentrated. Column chromatography is used to purify the crude product to obtain 1-[(6- bromohexanoyl)oxy]dodecan-3-yl octanoate (600 mg) in the form of a colorless liquid.

[0271] Step Four: Synthesis of 6-[(2-hydroxyethyl)({6-[(3-{[(pentadecan-8-yloxy)carbonyl]oxy}propionyl)oxy]hexyl})amino]hexyl 3-{[(pentadecan-8-yloxy)carbonyl]oxy}propanoate

[0272] Reaction Formula:

[0273] Material Ratio:

[0274] Procedure:

[0275] Dissolve 6-bromohexyl 3-(1-heptyloctoxycarbonyloxy)propanoate (500 mg, 0.99 mmol) in acetonitrile (10.0 mL), add ethanolamine (30.1 mg, 492 μmol), potassium carbonate (408 mg, 2.96 mmol), potassium iodide (188 mg, 1.13 mmol), tetrahydrofuran (5.00 mL), and acetonitrile (10.0 mL) successively, and then react at 75 °C for 12 hours under nitrogen protection. TLC is used to monitor the end of the reaction, and the reaction solution is filtered and concentrated. Column chromatography is used to purify the product to obtain 6-[6-[3-(1-heptyloctoxycarbonyloxy)propyl oxyl]hexyl-(2-hydroxyethyl)amino]hexyl 3-(1-heptyloctoxycarbonyloxy)propanoate (70.0 mg) in the form of a colorless liquid.

[0276] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.72-4.66 (m, 2H), 4.41-4.38 (t, J = 12 Hz, 4H), 4.12-4.09 (t, J = 12 Hz, 4H), 3.82 (s, 2H), 2.96-2.89 (m, 2H), 2.87 (s, 3H), 2.72-2.69 (t, J = 12 Hz, 4H) ppm.

[0277] LCMS: RT = 2.463, m / z 915.2 [M+H] + .

[0278] Preparation of compound 17

[0279] Step one: Synthesis of 6-bromohexyl 4-nitrophenyl carbonate

[0280] Reaction scheme:

[0281] Material ratio:

[0282] Operation process:

[0283] 6-bromohexan-1-ol (3.00 g, 16.5 mmol), 4-dimethylaminopyridine (4.05 g, 33.1 mmol) were dissolved in dichloromethane (20 mL), and the temperature was lowered to 0 °C. Chloroformate 4-nitrophenyl (4.01 g, 19.8 mmol) was dissolved in dichloromethane (10 mL) and added to the reaction solution at 0 °C. Finally, the reaction was carried out at 25 °C for 4 hours under nitrogen protection. TLC monitoring showed that a new spot was generated. The reaction solution was directly used in the next step.

[0284] Step two: Synthesis of benzyl 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoate

[0285] Reaction scheme:

[0286] Material ratio:

[0287] Operation process:

[0288] Benzyl 5-hydroxypentanoate (3.45 g, 16.5 mmol) was directly added to the reaction solution of the previous step, and the reaction was carried out at 25 °C for 12 hours. TLC monitoring showed that the reaction was completed. 50 mL of water was added, and dichloromethane was extracted twice, each time 50 mL. The organic phase was dried and concentrated. Column chromatography was used for purification to obtain benzyl 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoate (1.30 g) as a yellow liquid.

[0289] Step three: Synthesis of 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoic acid

[0290] Reaction scheme:

[0291] Material ratio:

[0292] Operation process:

[0293] Dissolve benzyl 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoate (1.00 g, 2.41 mmol) in methanol (10 mL), add palladium on carbon (512 mg, 0.481 mmol), under hydrogen atmosphere at 25 °C for 12 hours, TLC monitor the reaction is complete, filter, concentrate. Purify by column chromatography to give 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoic acid (530 mg) as a yellow liquid.

[0294] Step four: synthesis of pentadecan-8-yl 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoate

[0295] Reaction scheme:

[0296] Material ratio:

[0297] Operation process:

[0298] Dissolve 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoic acid (530 mg, 1.63 mmol), pentadecan-8-ol (409 mg, 1.79 mmol) in dichloromethane (6 mL), add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (156 mg, 0.814 mmol), 4-dimethylaminopyridine (19.9 mg, 0.162 mmol) in sequence, finally react under nitrogen atmosphere at 25 °C for 12 hours, TLC monitor the reaction has a new point, concentrate the reaction solution. Purify the crude product by column chromatography to give pentadecan-8-yl 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoate (830 mg) as a colorless liquid.

[0299] Step five: synthesis of pentadecan-8-yl 5-{[({6-[(2-hydroxyethyl)({6-[({[5-oxo-5-(pentadecan-8- oxy)pentyl]oxy}carbonyl)oxy]hexyl})amino]hexyl}oxy)carbonyl]oxy}pentanoate

[0300] Reaction scheme:

[0301] Material ratio:

[0302] Operation process:

[0303] Pentadecan-8-yl 5-{[({6-[(2-hydroxyethyl)({6-[({[5-oxo-5-(pentadecan-8- yloxy)pentyl]oxy}carbonyl)oxy]hexyl})amino]hexyl}oxy)carbonyl]oxy}pentanoate was prepared as a yellow liquid (40 mg) by purification by column chromatography from pentadecan-8-yl 5-({[(6-bromohexyl)oxy]carbonyl}oxy)pentanoate (600 mg, 1.12 mmol) dissolved in acetonitrile (4 mL) to which was added 2-aminoethan-1-ol (30.7 mg, 0.504 mmol), potassium carbonate (464 mg, 3.36 mmol), potassium iodide (278 mg, 1.68 mmol), tetrahydrofuran (2 mL) and finally reaction at 75 °C under nitrogen for 12 h, reaction monitored by TLC, filtered and concentrated.

[0304] 1 H NMR (400 MHz, CHLOROFORM-d) δ = 4.87 (t, J = 6.3 Hz, 2H), 4.17 - 4.10 (m, 8H), 3.85 (br d, J = 1.4 Hz, 1H), 3.01 - 2.80 (m, 5H), 2.38 - 2.29 (m, 4H), 1.79 - 1.67 (m, 18H), 1.65 - 1.48 (m, 36H), 1.46 - 1.35 (m, 10H), 1.35 - 1.20 (m, 44H), 0.88 (t, J = 6.8 Hz, 12H).

[0305] LCMS: RT = 2.174, m / z 970.7 [M+H] + .

[0306] Preparation of compound 6

[0307] Step one: synthesis of undecyl 6-bromohexanoate

[0308] Reaction scheme:

[0309] Material ratio:

[0310] Operation process:

[0311] Dissolve undecan-1-ol (3.00 g, 17.4 mmol) in dichloromethane (10.0 mL), add 6-bromohexanoic acid (5.09 g, 26.1 mmol), 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (5.01 g, 26.1 mmol), 4-dimethylaminopyridine (213 mg, 1.74 mmol) sequentially, and finally react at 25 °C for 12 hours under nitrogen protection. TLC monitoring shows that the reaction is completed, and the reaction solution is concentrated. Column chromatography of the crude product gives undecyl 6-bromohexanoate (6.00 g) as a colorless liquid.

[0312] Step Two: Synthesis of O1-[4-[2-hydroxyethyl-(6-oxo-6-undecyloxy-hexyl)amino]butyl] O4-(1- octylnonyl) butanedioate

[0313] Reaction Scheme:

[0314] Material Ratio:

[0315] Operation Process:

[0316] Dissolve O1-[4-(2-hydroxyethylamino)butyl] O4-(1-octylnonyl) butanedioate (1.00 g, 2.12 mmol) in acetonitrile (9.00 mL), add undecyl 6-bromohexanoate (889 mg, 2.54 mmol), potassium carbonate (1.03 mg, 7.42 mmol), potassium iodide (422 mg, 2.54 mmol), tetrahydrofuran (3.00 mL) sequentially, and finally react at 75 °C for 12 hours under nitrogen protection. TLC monitoring shows that the reaction is completed, filtration, and concentration. Purification by column chromatography gives O1-[4-[2-hydroxyethyl-(6-oxo-6-undecyloxy-hexyl)amino]butyl] O4-(1- octylnonyl) butanedioate (630 mg) as a colorless liquid.

[0317] 1 H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.86 (t, J=6.2 Hz, 1 H) 4.03 - 4.14 (m, 4 H) 3.68 (br t, J=5.0 Hz, 2 H) 2.73 - 2.80 (m, 2 H) 2.58 - 2.72 (m, 8 H) 2.31 (t, J=7.4 Hz, 3 H) 1.58 - 1.70 (m, 10 H) 1.51 (br d, J=6.0 Hz, 4 H) 1.22 - 1.36 (m, 42 H) 0.80 - 0.98 (m, 9 H) ppm.

[0318] LCMS: RT = 2.350, m / z = 741.0 [M+H] + .

[0319] Preparation of compound 8

[0320] Step one: Synthesis of 4-hexaoxo-4-oxalyl-butyric acid

[0321] Reaction scheme:

[0322] Material ratio:

[0323] Operation process:

[0324] Hexan-1-ol (3.00 g, 29.4 mmol) was dissolved in tetrahydrofuran (30 mL), tetrahydrofuran-2,5-dione (4.41 g, 44.0 mmol), 4-dimethylaminopyridine (3.59 g, 29.4 mmol), pyridine (3.48 g, 44.0 mmol) were added successively. Finally, the reaction was carried out at 60 °C for 12 hours under nitrogen protection, and the reaction was monitored by TLC. Concentration was carried out. The crude product was obtained as a colorless liquid (4-hexaoxo-4-oxalyl-butyric acid, 5.00 g) by column chromatography.

[0325] Step two: Synthesis of O4-(6-bromohexyl)O1-hexyl butanedioate

[0326] Reaction scheme:

[0327] Material ratio:

[0328] Operation process:

[0329] 4-hexaoxo-4-oxalyl-butyric acid (1.00 g, 4.90 mmol) was dissolved in dichloromethane (10.0 mL), 6-bromohexan-1-ol (1.34 g, 7.42 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.42 g, 7.42 mmol), 4-dimethylaminopyridine (66.0 mg, 500 μmol) were added successively. Finally, the reaction was carried out at 25 °C for 12 hours under nitrogen protection, and the reaction was monitored by TLC. Concentration was carried out. The crude product was obtained as a colorless liquid (O4-(6-bromohexyl)O1-hexyl butanedioate, 1.50 g) by column chromatography.

[0330] Step three: Synthesis of 4-(1-octyl nonoxo)-4-oxalyl-butyric acid

[0331] Reaction scheme:

[0332] Material ratio:

[0333] Procedure:

[0334] Dissolve 4-(l-octylnonoxo)-4-oxo-butyric acid (1.00 g, 2.80 mmol) in dichloromethane (10.0 mL), add 4-bromobutane-l-ol (0.64 g, 3.37 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.55 g, 4.21 mmol), 4-dimethylaminopyridine (34.0 mg, 280 μmol) successively, and finally react at 25 °C for 12 hours under nitrogen protection. TLC monitors the end of the reaction, and the reaction solution is concentrated. Column chromatography of the crude product gives colorless liquid 4-bromobutyl 4-(l-octylnonoxo)butyrate (1.10 g).

[0335] Step four: Synthesis of 4-bromobutyl 4-(l-octylnonoxo)butyrate

[0336] Reaction scheme:

[0337] Material ratio:

[0338] Procedure:

[0339] Dissolve 4-(l-octylnonoxo)-4-oxo-butyric acid (1.00 g, 2.80 mmol) in dichloromethane (10.0 mL), add 4-bromobutane-l-ol (0.64 g, 3.37 mmol), l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.55 g, 4.21 mmol), 4-dimethylaminopyridine (34.0 mg, 280 μmol) successively, and finally react at 25 °C for 12 hours under nitrogen protection. TLC monitors the end of the reaction, and the reaction solution is concentrated. Column chromatography of the crude product gives colorless liquid 4-bromobutyl 4-(l-octylnonoxo)butyrate (1.10 g).

[0340] Step five: Synthesis of 3-octyldodecanyloxy 6-[2-hydroxyethyl-[6-(3-octyldodecanoyloxy)-6-oxo-hexyl]amino]hexanoate

[0341] Reaction scheme:

[0342] Material ratio:

[0343] Procedure:

[0344] Dissolve 4-bromobutyl 4-(l-octylnonoxyl)butylate (3.00 g, 6.28 mmol) in acetonitrile (10.0 mL), add ethanamine (4.60 g, 75.4 mmol) sequentially, and finally potassium carbonate (1.03 g, 7.42 mmol) and potassium iodide (422 mg, 2.54 mmol) in tetrahydrofuran (3.00 mL) under nitrogen at 25 °C for 12 hours. Monitor the reaction by TLC until completion, and concentrate. Purify by column chromatography to obtain O1-[4-(2- hydroxyethylamino)butyl] O4-(l-octylnonyl)butanedioate (2.40 g) as a colorless liquid.

[0345] Step six: Synthesis of O1-hexyl O4-[6-[2-hydroxyethyl-[4-[4-(l-octylnonoxyl)-4- oxo-oxo-butyryl]oxybutyl]amino]hexyl] butanedioate

[0346] Reaction Scheme:

[0347] Material ratio:

[0348] Operation process:

[0349] Dissolve O1-[4-(2-hydroxyethylamino)butyl] O4-(l-octylnonyl)butanedioate (1.00 g, 2.12 mmol) in acetonitrile (9.00 mL), add O4-(6-bromohexyl) O1-hexyl butanedioate (930 mg, 2.54 mmol), potassium carbonate (1.03 g, 7.42 mmol), potassium iodide (422 mg, 2.54 mmol), and tetrahydrofuran (3.00 mL) sequentially under nitrogen at 75 °C for 12 hours. Monitor the reaction by TLC until completion, filter, and concentrate. Purify by column chromatography to obtain O1-hexyl O4-[6-[2-hydroxyethyl-[4-[4-(l-octylnonoxyl)-4-oxo-oxo-butyryl]oxybutyl]amino]hexyl] butanedioate (880 mg) as a colorless liquid.

[0350] 1 H NMR (400 MHz, CHLOROFORM-d) δ ppm 4.86 (t, J=6.2 Hz, 1 H) 3.95-4.17 (m, 6 H) 3.67 (br t, J=4.8 Hz, 2 H) 2.70-2.80 (m, 2 H) 2.50-2.70 (m, 12 H) 1.45-1.74 (m, 14 H) 1.16-1.42 (m, 34 H) 0.59-0.97 (m, 9 H) ppm.

[0351] LCMS: RT = 2.450, m / z = 756.5. [M+H] + .

[0352] Example 1 Preparation and testing of lipid nanoparticles (LNP)

[0353] To verify whether the Lipid Nanoparticle (LNP) formulations made from the ionizable lipid compounds disclosed in this application can effectively encapsulate mRNA and maintain the structural integrity of mRNA. The prepared ionizable lipid compounds, distearoylphosphatidylcholine (DSPC, purchased from NOF Corporation, item number: S01005), cholesterol (purchased from NOF Corporation, item number: O01001) and dimyristoylglycerol-polyethylene glycol 2000 (DMG-PEG2000, purchased from Genview Biotech, item number: O02005) were dissolved in ethanol (manufacturer: Nanjing Chemical Reagent Co., Ltd., purity 99.6%) solution, respectively, and then mixed according to a certain molar ratio to prepare a mixed lipid ethanol solution, in which the total concentration of lipids was 12.5 mM (the unit of measurement “M” appearing in this application means mol / L). The self-prepared 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 at 12 mL / min, and the volume ratio of the mixed lipid ethanol solution to the mRNA solution prepared in the previous step was controlled at 1:3, and the nitrogen-phosphorus ratio of the ionizable lipid to mRNA was controlled at 3-15:1 to prepare the lipid nanoparticles. 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 concentrated by ultrafiltration (manufacturer: Amicon-Ultra, molecular weight cutoff: 10 kDa) to obtain the LNP formulation in which the Fluc mRNA was encapsulated by the ionizable lipid described in this application and DSPC, cholesterol and DMG-PEG2000. The particle size and polydispersity index (PDI) of each LNP formulation were determined by dynamic light scattering method using a Malvern Zetasizer Ultra instrument (manufacturer: Malvern); the encapsulation efficiency of LNP was determined using the Quant-it Ribogreen RNA quantification assay kit (manufacturer: ThermoFisher Scientific, item number: R11490).

[0354] Table 1

[0355] In the art, a PDI less than 0.3 indicates that the nanoparticle size in the LNP formulation is relatively uniform; the encapsulation efficiency is used to indicate whether the LNP can effectively encapsulate mRNA, and an encapsulation efficiency higher than 70% indicates that the LNP can effectively encapsulate mRNA. The LNP prepared in this application has a particle size of 60-100 nm, a PDI less than 0.15, and an encapsulation efficiency higher than 90%.

[0356] Example 2 Animal in vivo study of LNP formulations

[0357] In this example, the LNP prepared in Example 1 was injected into 6-8 week old female Balb / C mice (Vital River) via tail vein injection or lower limb muscle injection at a dose of 5 pg per mouse (n = 3, i.e. 3 mice were used for injection and testing in each group, and the data presented is the average of each group), and D-luciferin potassium salt was injected intraperitoneally at specific time points (4h, 24h, 48h in this example) after administration, and then the luminescence was detected by IVIS Spectrum small animal live imaging instrument (manufacturer: PerkinElmer), and the total luminescence intensity of the mouse live expression site (such as liver, lower limb administration site, etc.) was counted, and the higher the luminescence intensity, the higher the luciferase expression, i.e. the better the corresponding LNP formulation expressed in mice. The total luminescence intensity is measured by bioluminescence imaging, and the luminescence intensity data of the luminescent site is obtained after 6-15 minutes (min) of intraperitoneal injection of D-luciferin potassium salt. The total luminescence intensity of the live expression area is counted by Living Image software (manufacturer: PerkinElmer). Generally, the total luminescence intensity of the mouse detected by the live imaging instrument without administration treatment is in the order of 10 5 .

[0358] Referring to the above mouse in vivo experimental method, the LNP formulation in Example 1 was injected into 6-8 week old female Balb / C mice via tail vein injection at a dose of 5 pg per mouse, and the total luminescence intensity of the liver area was counted, and the test results are shown in Table 2 and Figure 1. The LNP formulations tested in this example all have strong expression in mice, and the AUC range is 10 9 ~ 10 12 , indicating that the ionizable lipids described in the preparation example can effectively deliver mRNA to the body and express. Overall expression trend: compound 3 > compound 2.

[0359] Table 2 Area under the curve (AUC) of 4-48h expression kinetics in liver area

[0360] Similarly, the LNP formulation prepared in Example 1 was injected into 6-8 week old female Balb / C mice via muscle injection (right lower limb of calf) at a dose of 5 pg per mouse, and the total luminescence intensity of the liver area and the administration muscle site was counted and the corresponding 4-48h expression kinetics curve area (AUC) was calculated, and the test results are shown in Table 3 and Figures 2-3. It can be seen that the intramuscular injection of the liver area and the administration site expresses: compound 3 > compound 2.

[0361] Table 3 Area under curve (AUC) of expression kinetics in liver region and administration site 4-48h

[0362] Although the specific embodiments of the present application are described above, those skilled in the art should understand that this is only an example, the protection scope of the present application is defined by the appended claims. Those skilled in the art can make various changes or modifications to these embodiments without departing from the principles and essence of the present application, and these changes and modifications all fall within the protection scope of the present application.

[0363] Example 3 Preparation and detection of lipid nanoparticles (LNP)

[0364] In this example, compound 4 and compound 5 were selected as ionizable lipids, and LNP formulations (encapsulating Fluc mRNA) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 4, and in the manner of Reference Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used during dialysis. The particle size, PDI and encapsulation efficiency of all LNP formulations in this example were determined. As can be seen from Table 4, the particle size of the LNP formulations prepared in this example was between 60-110 nm, the PDI was less than 0.3, and the encapsulation efficiency was higher than 90%.

[0365] Table 4

[0366] Example 4 In vivo study of LNP formulations

[0367] According to the in vivo test method of Reference Example 2, the LNP reagent prepared in Example 3 was injected into 6-8 week old female Balb / C mice through the tail vein at a dose of 5 μg per mouse, and the total luminescence intensity of the mouse liver in vivo was counted. The test results are shown in Table 5 and Figure 4, and the LNP formulations prepared in this example have strong expression in mice.

[0368] Table 5 Area under curve (AUC) of expression kinetics in liver region 4-48h

[0369] Example 5 Preparation and detection of lipid nanoparticles (LNP)

[0370] The compounds 7 and 9 were selected as ionizable lipids, and the LNP formulations (encapsulating Fluc mRNA) were prepared according to the molar ratios and nitrogen to phosphorus ratios in Table 6 by referring to the method of Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetate solution with a pH of 7.5 was used during dialysis. The particle size, PDI and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 6, the particle size of the LNP formulations prepared in this example was between 60-100 nm, the PDI was less than 0.25, and the encapsulation efficiency was higher than 90%.

[0371] Table 6

[0372] Example 6 Animal in vivo study of LNP formulations

[0373] According to the method of the in vivo test of mice in Reference Example 2, the LNP reagent prepared in Example 5 was injected into 6-8 week old female Balb / C mice through the tail vein at a dose of 5 μg per mouse, and the total luminescence intensity of the liver of the mice in vivo was counted. The test results are shown in Table 7 and FIG. 5, and the LNP formulations prepared in this example have strong expression in mice.

[0374] Table 7 Area under the curve (AUC) of expression kinetics in liver area 4-48 h

[0375] Example 7 Preparation and detection of lipid nanoparticles (LNP)

[0376] The compound 1 was selected as an ionizable lipid, and the LNP formulations (encapsulating Fluc mRNA) were prepared according to the molar ratios and nitrogen to phosphorus ratios in Table 8 by referring to the method of Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetate solution with a pH of 7.5 was used during dialysis. The particle size, PDI and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 8, the particle size of the LNP formulations prepared in this example was between 60-140 nm, the PDI was less than 0.25, and the encapsulation efficiency was higher than 85%.

[0377] Table 8

[0378] Example 8 Animal in vivo study of LNP formulations

[0379] According to the method of the in vivo test of mice in Reference Example 2, the LNP reagent prepared in Example 7 was injected into 6-8 week old female Balb / C mice through the tail vein at a dose of 5 μg per mouse, and the total luminescence intensity of the liver of the mice in vivo was counted. The test results are shown in Table 9 and FIG. 6, and the LNP formulations prepared in this example have strong expression in mice.

[0380] Table 9 Area under curve (AUC) of expression kinetics of liver region 4-48h

[0381] Example 9 Preparation and detection of lipid nanoparticles (LNP)

[0382] The compounds 10, 11, 12, 14 and 17 were selected as ionizable lipids in this example, and LNP formulations (encapsulating Fluc mRNA) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 10, in the manner of Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetic acid solution with a pH of 7.5 was used for dialysis. The particle size, PDI and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 10, the particle size of the LNP formulations prepared in this example was between 60-140 nm, the PDI was less than 0.25, and the encapsulation efficiency was higher than 85%.

[0383] H23 and H21 are cited from international application WO2023186041A1, and were synthesized using the same preparation method as WO2023186041A1.

[0384] Table 10

[0385] Example 10 In vivo study of LNP formulations in animals

[0386] According to the in vivo test method of Example 2, the LNP reagent prepared in Example 7 was injected into the tail vein of 6-8 week old female Balb / C mice at a dose of 5 μg per mouse, and the total luminescence intensity of the mouse liver in vivo was counted. The test results are shown in Table 11 and Figure 7, and the LNP formulations prepared in this example have strong expression in mice.

[0387] Table 11 Area under curve (AUC) of expression kinetics of liver region 4-48h

[0388] Example 11 Preparation and detection of lipid nanoparticles (LNP)

[0389] The compounds 6 and 8 were selected as ionizable lipids, and the LNP formulations (encapsulating Fluc mRNA) were prepared according to the molar ratio and nitrogen-phosphorus ratio in Table 12 in the manner of Reference Example 1. The mRNA in this example was diluted in a 25 mM sodium acetate solution with a pH of 5.0, and a 20 mM Tris-acetate solution with a pH of 7.5 was used during dialysis. The particle size, PDI and encapsulation efficiency of all LNP formulations in this example were determined. As shown in Table 12, the particle size of the LNP formulations prepared in this example was between 60-140 nm, the PDI was less than 0.35, and the encapsulation efficiency was higher than 84%.

[0390] Table 12

[0391] Example 12 In vivo study of LNP formulations in animals

[0392] According to the in vivo test method of mice in Reference Example 2, the LNP reagent prepared in Example 11 was injected into the tail vein of 6-8 week old female Balb / C mice at a dose of 5 μg per mouse, and the total luminescence intensity of the liver in vivo of the mice was counted. The test results are shown in Table 13 and Figure 8, and the LNP formulations prepared in this example have strong expression in mice.

[0393] Table 13 Area under the curve (AUC) of expression kinetics in liver area 4-48 h

Claims

1. A compound I or a pharmaceutically acceptable salt thereof: in, R 1 C is a C that is substituted with one or more hydroxyl groups 1-6 alkyl; X and Y are independently C 3-12 Alkylene; Z 1 and Z 2 Independently W 1 For chemical bonds, * indicates that it is related to Z 1 connect; W 2 for * indicates that it is related to Z 2 connect; R w11 R w12 R w21 and R w22 Independently for C 1-5 Straight-chain alkylene; R 2 C 1-24 alkyl; R 3 C 1-24 alkyl.

2. The compound I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1)R 1 C replaced by a hydroxyl group 1-6 alkyl; (2)R 1 In this case, only the terminal carbon atom at the end that is not bonded to the N atom is replaced by a hydroxyl group; (3) X and Y are independently C 3-7 Alkylene; (4) X and Y are independently straight-chain alkylene groups; (5)Z 1 for * indicates W 1 connect; (6)Z 2 for * indicates W 2 connect; (7)R w11 R w12 R w21 and R w22 C independently 2-4 Straight-chain alkylene; (8)R 2 C is a straight chain or has one branch 4-18 alkyl; and (9)R 3 for R 31 and R 32 C independently 3-10 alkyl; Preferably, the compound I or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) X and Y are independently C 3-6 Alkylene; (2)R 2 C 5-11 Straight-chain alkyl or C with one branch 15-17 alkyl; and (3)R 31 and R 32 C independently 6-9 Straight-chain alkyl groups; for example 3. The compound I as described in claim 2, or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1)R 1 for (2) X and Y are independently... (3)W 1 For chemical bonds, * indicates that it is related to Z 1 connect; (4)W 2 for * indicates that it is related to Z 2 connect; (5)R 2 for and (6)R 3 for Preferably, the compound I or a pharmaceutically acceptable salt thereof satisfies one or more of the following conditions: (1) for and (2) for 4. Compound I or a pharmaceutically acceptable salt thereof as described in any one of claims 1-3, characterized in that, Compound I has a structure as shown in IA, IB, or IC: Among them, R 1 X, Y, Z 1 W 1 R 2 R w21 R w22 and R 3 The definition is as described in any one of claims 1-3.

5. The compound I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, Compound I has any of the following structures:

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

7. The composition according to claim 6, characterized in that, The composition is a lipid carrier; Preferably, the lipid carrier satisfies one or more of the following conditions: (1) The lipid carrier further includes a diluent, which may be a phosphate buffer, a sodium acetate buffer or a Tris-acetic acid buffer. (2) The lipid carrier further includes phospholipids, which may be phospholipid molecules having an electric polar end and a nonpolar end of a fatty chain, preferably distearylphosphatidylcholine, myristoyl phosphocholine, dioleoyl phosphocholine, palmitoyl phosphocholine, docosyl phosphocholine or palmitoyl phosphocholine, for example, distearylphosphatidylcholine. (3) The lipid carrier further includes PEG lipids, which may be lipid molecules modified with a hydrophilic end of polyethylene glycol; 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. The lipid carrier described in (4) further includes sterols, which may be animal, plant or fungal sterols, preferably selected from one or more of cholesterol, sitosterol, ergosterol, campesterol, stigmasterol, brassosterol, tomatine, ursolic acid and α-tocopherol, such as cholesterol; More preferably, the lipid carrier satisfies one or more of the following conditions: (1) The molar ratio of substance Z to sterol is 0.5-5:1, preferably 0.5-3:1, for example 1.3:1; (2) The molar ratio of substance Z to phospholipid is 1-15:1, preferably 2-8:1, for example 3-6:1; (3) The molar ratio of substance Z to 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 substance Z is 30 mol% to 70 mol%, for example 50 mol%; (6) The molar content of the phospholipid is from 5 mol% to 20 mol%, for example, 10 mol%; (7) The molar content of the sterol is from 20 mol% to 60 mol%, for example 38.5 mol%; (8) The molar content of the PEG lipid is approximately 0.2 mol% to 5 mol%, for example, 1.5% mol; The lipid carrier described in (9) comprises the substance Z, the diluent, the phospholipid, the PEG lipid, and the sterol.

8. The use of compound I as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof, or a composition as described in any one of claims 6-7, in the preparation of a nucleic acid prophylactic agent and / or therapeutic agent delivery carrier; The nucleic acid prophylactic agent 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 more 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 includes nucleic acid prophylactic agents and / or therapeutic agents, as well as compositions as described in any one of claims 6-7; The nucleic acid prophylactic agent 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 more 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 nanoparticles as described in 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 (3-16):1, for example 6:1; (2) The lipid nanoparticles have a particle size of 10-200 nm, preferably 40-150 nm, such as 50.32 nm, 53.71 nm, 59.16 nm, 61.74 nm, 67.40 nm, 84.31 nm, 63.89 nm, 95.80, 100.27, 73.07, 62.60, 97.72, 131.87 or 131.40; or for example 84.31 nm, 63.89 nm, 95.80, 100.27, 73.07, 62.60 or 131.40; (3) The polydispersity index of the lipid nanoparticles is 0.001-0.3; for example, 0.135, 0.323, 0.107, 0.089, 0.106, 0.188, 0.201, 0.129, 0.079, 0.159, 0.298, 0.235 or 0.073, or for example, 0.129, 0.079, 0.159, 0.298, 0.235 or 0.073; (4) The encapsulation efficiency of the lipid nanoparticles is 84%-100%, for example, 84.5%, 92.5%, 93.4%, 96.3%, 94.9%, 94.1%, 92.9%, 96.4%, 96.7%, 92.7%, 93.5%, 96.2%, 96.1% or 86.8%; preferably, the encapsulation efficiency of the lipid nanoparticles is 85%-100%, for example, 96.40% or 96.66%, 92.74%, 93.51%, 96.17%, 96.10% or 86.82%; The composition described in (5) encapsulates the nucleic acid preventive agent and / or therapeutic agent.

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