CN116655486A - Long-chain alkyl ester amine compound and its preparation method and application in nucleic acid delivery - Google Patents

Abstract

The present invention relates to a long-chain alkyl ester amine compound represented by formula (I) and its preparation method and application in nucleic acid delivery. The long-chain alkyl ester amine compound provided by the present invention is used as a lipid molecule in the delivery of disease treatment Or preventive agents have excellent encapsulation efficiency and delivery effect, while exhibiting low liver and kidney toxicity and relatively selective distribution characteristics, providing more selection basis for the delivery of disease treatment or preventive agents.

Description

Long-chain alkyl ester amine compound and its preparation method and its application in nucleic acid delivery application

This application is a divisional application of a Chinese invention patent application (application number: 202210546254.8, application date: May 19, 2022, invention name: long-chain alkyl ester amine compounds and their preparation methods and applications in nucleic acid delivery) .

technical field

The invention belongs to the technical field of medicine, in particular, the invention relates to a long-chain alkyl ester amine compound, a preparation method thereof and an application in nucleic acid delivery.

Background technique

The nucleic acid synthesized by in vitro transcription technology is transported into the tissue cells by means of the ester nanoparticle delivery system, and the target protein is translated by the non-translational system of the own cell. With the driving function, the purpose of treatment is finally achieved. Nucleic acid-based therapeutic technology has the advantages of rapid preparation, low cost, and safety, which makes it stand out among many therapeutic methods and is widely used in the fields of cancer, infectious diseases, and rare diseases. However, since the nucleic acid itself is unstable and easily degraded in vivo, the selection of a stable delivery system is the key to the development of this type of drug.

With the vigorous development of nanoliposome technology, researchers are committed to developing new synthetic lipids and improving the drug-loading capacity of liposomes, so cationic lipids came into being for the effective transfer of negatively charged drugs, especially The delivery of nucleic acid drugs has attracted much attention. Cationic lipids generally have a positively charged head group connected to a hydrophobic tail segment (cholesterol or aliphatic chain) through a linkage (amide, ester, ether bond), and its structure is an important factor determining the efficacy of nucleic acid drugs. In recent years, lipid molecules have been greatly developed, from permanently charged cationic lipid molecules, such as DOTAP, DOGS (structural formula shown below), etc., to ionizable cationic lipid Dlin-DMA. On the basis of DLin-DMA, MC3 (structural formula is shown below) was obtained by modifying the chain length and substitution position. The world's first siRNA drug, Onpattro, was approved for marketing in 2018 for the treatment of nerve damage caused by thyroxin amyloidosis. The key lipid molecule used in this drug is MC3. On the basis of MC3, Moderna Biotech optimized the lipid molecule SM-102 (structural formula shown below) with hydroxyethyl group attached to the nitrogen atom, and applied this compound in the development of COVID-19 vaccine, which will be launched in 2020 Received FDA authorization on December 8.

Although some cationic delivery molecules have been used for the delivery of disease treatment or prevention agents, there are still some problems in the delivery of nucleic acids, such as unsatisfactory stability and selectivity. Therefore, it is particularly important to invent new delivery molecules.

Contents of the invention

One of the objects of the present invention is to provide a series of long-chain alkyl ester amine compounds.

Another object of the present invention is to provide a preparation method for long-chain alkyl ester amines.

Another object of the present invention is to provide a composition containing long chain alkyl ester amines.

Another object of the present invention is to provide the application of long-chain alkyl ester amine compounds as lipid molecules in nucleic acid delivery.

Definition of Terms

The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The nomenclature used herein and the laboratory procedures in organic chemistry, medicinal chemistry, and biology described herein are those well known and commonly used in the art. Unless mentioned otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this art belongs.

As used in the description of embodiments of the present invention and the appended claims, the singular forms "a", "an", "the" and "it" are used to refer to both the singular and the plural of the article, unless the context clearly dictates otherwise Reference. For example, a compound includes one or more than one compound.

As used herein, "and/or" means and includes any and all possible combinations of one or more of the associated listed items.

As used herein, the term "disease" or "illness" refers to any change in the state of the body or some organ that interrupts or interferes with the performance of its function and/or causes symptoms.

As used herein, the term "treatment" aims at alleviating or eliminating the targeted disease state or disorder. If a subject receives a therapeutic amount of a biomacromolecule or a small chemical molecule according to the methods described herein, the subject exhibits an observable and/or detectable reduction in one or more signs and symptoms or improvement, the subject was successfully "treated". It should also be understood that reference to treatment of a disease state or disorder includes not only complete treatment, but also incomplete treatment while achieving some biologically or medically relevant result.

Technical topic one

The present invention provides long-chain alkyl ester amine compounds having the structure shown in formula I:

Wherein, X is a straight chain alkane of C5-C12.

In some preferred embodiments of the invention, the compounds include the following structures:

Technical Topic II

The present invention also provides a synthetic method for the compound shown in formula I, comprising the following steps:

Step 1: Add OH-X-NH ₂ and DIEA to the solution of compound A in a protic solvent, heat and react for 12 to 36 hours, dilute with water, extract with ethyl acetate, and purify to obtain compound B;

Step 2: Add compound B, potassium carbonate and potassium iodide to the solution of compound C in an aprotic solvent, heat and react for 24-48 hours, dilute with water, extract with ethyl acetate, and purify to obtain the compound of formula I.

Further, the solvent in step 1 is ethanol or propanol.

Further, the solvent in step 2 is a mixed solvent of 2-methyltetrahydrofuran and acetonitrile, or a mixed solvent of cyclopentyl methyl ether and acetonitrile, and the ratio of 2-methyl tetrahydrofuran or cyclopentyl methyl ether to acetonitrile is 1 :1 to 3:1.

Further, the purification is purified by silica gel column chromatography.

Technical Topic Three

The present invention provides a composition including a therapeutic or preventive agent and a carrier for delivering the therapeutic or preventive agent, and the carrier includes the long-chain alkyl ester amine compound described in Technical Subject 1.

Further, the therapeutic or preventive agent includes one or more of nucleic acid molecules, proteins, polypeptides or small molecule compounds.

Further, the nucleic acid includes any form of nucleic acid molecule, including but not limited to single-stranded DNA, double-stranded DNA, short isomers, agomir, antagomir, antisense molecules, small interfering RNA (siRNA), asymmetric interfering RNA ( aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA) and other forms of RNA molecules known in the art, or locked nucleic acids (LNA), peptide nucleic acid (PNA) and morpholine oligonucleotides and other nucleic acid mimics.

Further, the nucleic acid is selected from at least one mRNA encoding an antigen or a fragment or epitope thereof.

Further, the therapeutic or preventive agent is a vaccine.

Further, the small molecular compound may be selected from antineoplastic drugs, anti-infective drugs, antidepressants, anticonvulsants, antibiotics/antibacterial agents, antifungal drugs, antiparasitic drugs, immunomodulators or anesthetics.

Further, the "pharmaceutical composition" may also contain other excipients, such as: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents , solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, taste-masking agents, colorants, anti-caking agents, humectants, chelating agents, plasticizers, tackifiers, antioxidants, preservatives agents, stabilizers, surfactants and buffers.

The compound of the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

Technical Topic Four

The present invention also provides the application of the compound represented by formula I in the preparation of nucleic acid drugs, vaccines, protein or polypeptide drugs, and small molecule drugs.

Further, the application is in the preparation of nucleic acid delivery drugs.

Further, the application is in the preparation of mRNA medicines.

Further, the application is in the preparation of mRNA vaccines.

The beneficial effects of the present invention are as follows:

The present invention provides a new class of long-chain alkyl ester amine compounds. Through extensive research and experimental verification, it is found that this type of long-chain alkyl ester amine compound has high encapsulation efficiency, good delivery effect, low hepatotoxicity and relative The characteristics of selection distribution provide more selection basis for the delivery of disease treatment or prevention agents.

Description of drawings

In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings that need to be used in the description of the specific embodiments or the prior art.

Figure 1 shows the distribution of delivered mRNA in vivo.

Detailed ways

The present invention is described below in conjunction with specific examples. These examples are not intended to limit the scope of the present invention, but to provide guidance for those skilled in the art to prepare and use the compounds and compositions of the present invention. The chemical names of the compounds described in this application were generally generated from ChemDraw Ultra (ChambridgeSoft) and/or generally followed the principles of IUPAC nomenclature.

The synthetic route of the compound of this embodiment part is as follows:

Example 1

Add 5-amino-1-pentanol (88mg, 0.86mmol) and DIEA (213μL, 1.29mmol) to compound A (200mg, 0.43mmol) in ethanol (5mL) solution, heat to reflux for 20 hours, TLC monitors the reaction, the reaction Cool to room temperature when complete. Add an equal volume of water to the reaction solution for dilution, extract with ethyl acetate (10mL×3), concentrate, and perform silica gel column chromatography (dichloromethane:methanol=10:1) to obtain compound B-1, X=(CH ₂ ) ₅ (172 mg, yield 82.69%).

To the mixed solvent of compound B-1 (260mg, 0.54mmol) in acetonitrile and cyclopentyl methyl ether (cyclopentyl methyl ether: acetonitrile = 2:1, 3mL), add 5 times the equivalent of compound C (936mg, 2.69 mmol), K ₂ CO ₃ (224mg, 1.62mmol) and KI (90mg, 0.54mmol), heated at 90°C for 24 hours, monitored by TLC, and cooled to room temperature after the reaction was complete. The reaction solution was diluted with water, extracted with ethyl acetate (10mL×3), concentrated, and subjected to silica gel column chromatography (dichloromethane:methanol=20:1) to obtain oily compound 1 (140mg, yield 34.5%), ¹ H NMR (500MHz, CDCl ₃ ) δ4.88–4.83(m,1H), 4.05(t, J＝6.7Hz, 2H), 3.63(t, J＝6.3Hz, 2H), 2.85–2.58(brs, 6H) ¹³ C NMR (126MHz, CDCl ₃ )δ173.77, 173.60, 74.23, 64.61, 62.13, 53.26, 53.11, 34.64, 34.17, 34.11, 32.07, 31.94, 31.90, 31.88, 29.63, 29.56, 29.37, 29 .29, 29.27, 29.19, 29.08, 29.02, 28.69, 27.08, 26.75, 25.97, 25.36, 25.02, 24.81, 24.61, 23.53, 22.72, 22.70, 14.14; MS-ESI (m/z): 752 (M+H) ⁺ .

Example 2

The preparation method is the same as that of compound 1, using 6-amino-1-hexanol as a raw material to obtain oily compound 2, ¹ H NMR (500MHz, CDCl ₃ ) δ4.86–4.81(m,1H), 4.04(t,J= 6.8Hz, 2H), 3.63(t, J＝6.4Hz, 2H), 2.99–2.94(brs, 6H), 2.29(dt, J＝23.7, 4H), 1.87–1.75(m, 7H), 1.69–1.55 (m,8H),1.51–1.24(m,55H),0.86(t,J=6.8Hz,9H); ¹³ C NMR(151MHz,CDCl ₃ )δ173.58,173.44,74.37,64.80,62.41,52.57,52.49, 52.47, 34.61, 34.23, 33.92, 32.29, 32.01, 31.97, 29.70, 29.63, 29.61, 29.43, 29.37, 29.35, 29.00, 28.83, 28.74, 26.79, 26.55, 26.45, 26.03, 2 5.43, 25.17, 24.96, 24.32, 23.25, 23.19, 22.78, 14.22; MS-ESI (m/z): 766 (M+H) ⁺ .

Example 3

The preparation method is the same as that of compound 1, using 7-amino-1-heptanol as a raw material to obtain oily compound 3, ¹ H NMR (500MHz, CDCl ₃ ) δ4.83–4.81(m,1H), 4.02(t,J= 6.8Hz,2H),3.59(t,J＝6.6Hz,2H),2.57(brs,6H),2.28–2.22(m,4H),1.81(brs,1H),1.62–1.47(m,15H), 1.30–1.22(m,56H),0.84(t,J＝6.8Hz,9H); MS-ESI(m/z):780(M+H) ⁺ .

Example 4

The preparation method is the same as that of compound 1, using 8-amino-1-octanol as the raw material to obtain oily compound 4, ¹ H NMR (500MHz, CDCl ₃ ) δ4.85–4.80(m,1H), 4.02(t,J= 6.8Hz,2H),3.59(t,J＝6.6Hz,2H),2.58(brs,6H),2.28–2.22(m,4H),1.81(brs,1H),1.64–1.46(m,15H), 1.29–1.22(m,58H),0.84(t,J=6.8Hz,9H); ¹³ C NMR(126MHz,CDCl ₃ )δ173.71,173.63,74.21,64.58,62.84,53.53,34.69,34.20,32.79,31.96, MS-ESI(m/ z):794(M +H) ⁺ .

Example 5

The preparation method is the same as that of compound 1, using 9-amino-1-nonanol as a raw material to obtain oily compound 5, ¹ H NMR (500MHz, CDCl ₃ ) δ4.85–4.81(m,1H), 4.02(t,J= 6.8Hz,2H),3.59(t,J＝6.6Hz,2H),2.56(brs,6H),2.28–2.21(m,4H),1.81(brs,1H),1.62–1.47(m,15H), 1.31–1.20(m,60H),0.84(t,J＝6.8Hz,9H); MS-ESI(m/z):808(M+H) ⁺ .

Example 6

The preparation method is the same as that of compound 1, using 10-amino-1-n-decyl alcohol as a raw material to obtain oily compound 6, ¹ HNMR (500MHz, CDCl ₃ ) δ4.86–4.81(m,1H), 4.03(t,J= 6.8Hz, 2H), 3.60(q, J=6.2Hz, 2H), 2.39(s, 6H), 2.29–2.23(m, 4H), 1.72–1.69(m, 1H), 1.64–1.56(m, 6H ),1.55–

1.52(m,1H),1.51–1.40(m,7H),1.34–1.23(m,63H),0.85(t,J＝6.8Hz,9H); ¹³ CNMR(126MHz,CDCl ₃ )δ173.89,173.69,74.17 ,64.51,62.95,54.14,53.96,34.78,34.41,34.23,32.91,31.98,31.94,29.68,29.66,29.63,29.61,29.58,29.51,29.41,29.36,29.34,29.31, 29.29, 28.74, 27.65, 27.55, 27.23 ,26.01,25.86,25.39,25.20,25.03,22.75,22.73,14.17; MS-ESI(m/z):822(M+H) ⁺ .

Example 7

The preparation method is the same as that of compound 1, using 12-amino-1-dodecanol as a raw material to obtain oily compound 7, ¹ HNMR (500MHz, CDCl ₃ ) δ4.85–4.80 (m, 1H), 4.02 (t, J = 6.8Hz, 2H), 3.59(t, J=6.6Hz, 2H), 2.51(brs, 6H), 2.28–2.23(m, 4H), 1.63–1.57(m, 5H), 1.54–1.45(m, 9H ),1.33–1.22(m,68H),0.85(t,J=6.7Hz,9H); ¹³ C NMR(151MHz,CDCl ₃ )δ173.77,173.66,74.19,64.56,62.97,53.74,34.71,34.28,34.21, 32.89, 31.97, 31.93, 29.66, 29.64, 29.59, 29.57, 29.49, 29.39, 29.32, 29.30, 29.22, 29.20, 28.72, 27.48, 27.36, 27.03, 26.00, 25.84, 25.38, 2 5.12, 24.85, 22.74, 22.73, 14.17; MS-ESI(m/z):850(M+H) ⁺ .

The preparation of embodiment 8 reporter gene luciferase mRNA

1.1 Plasmid linearization

The reporter gene plasmid pUC57-luc contains T7 promoter, 5'UTR, luciferase sequence, 3'UTR and polyA tail, and there is a SapI restriction site behind the last A in the polyA tail. 10 μg of the plasmid to be tested, 1 μL of the restriction enzyme Sap I (10000 U/mL), 5 μL of 10xCutsmart buffer, and 50 μL was supplemented with ddH2O. Restriction enzyme Sap I and 10xCutsmart buffer are complementary products, NEB, catalog number R0569L. The reaction conditions are 37°C, 3h. After the reaction was completed, 2 μL of the digested product was taken and subjected to 1% agarose gel electrophoresis to detect the linearization of the plasmid. After confirming that the restriction enzyme digestion was completed, the linearized plasmid was recovered and purified using the Rapid DNA Product Purification Kit (Converse Century, CW2301M).

1.2 In vitro transcription

Using the linearized plasmid as a template for in vitro transcription, high-yield T7 RNA transcription kit was used for in vitro transcription. High Yield T7 RNA Transcription Kit, the product name is High Yield T7 RNA Synthesis Kit, Shanghai Zhaowei Technology Development Co., Ltd., the catalog number is ON-040; 5×Reaction Buffer, 100mM ATP Solution, 100mM CTP Solution, 100mM GTP Solution , Enzyme mix, DNase I, Ammonium AcetateStop Solution, Lithium Chloride(LiCl) Precipitation Solution are all components in the high-yield T7 RNA transcription kit. 100mMΨUTP Solution, full name N1-Me-pUTP, 100mM, Shanghai Zhaowei Technology Development Co., Ltd., catalog number R5-027.

Specific steps for in vitro transcription: first, prepare a reaction system, mix well and react at 37°C for 3 hours; then, add 1 μL DNase I (1U content) and react at 37°C for 15 minutes; then add 15 μL Ammonium Acetate Stop Solution.

Reaction system: 5×Reaction Buffer 4μL, 100mM ATP Solution 2μL, 100mMΨUTPSolution 1μL, 100mM CTP Solution 2μL, 100mM GTP Solution 2μL, Enzyme mix 2μL, linearized plasmid (DNA content 500ng-1μg), Nuclease-free H2O Qi Zhi 20 μL. Table 2 In vitro transcription system

1.3 RNA purification

Add 1/3 volume of 7.5M LiCl to the in vitro transcription reaction system (to make the final concentration 2.5M), and place at -20°C for 30min. Centrifuge at 12000g for 15min, the RNA precipitates at the bottom, discard the supernatant. Add 1 mL of 70% ethanol to wash the RNA, centrifuge at 12000 g for 5 min, and discard the supernatant. After drying, add 50 μL of RNase-free water to dissolve the precipitate, and use a UV spectrophotometer to quantify mRNA to obtain capped in vitro transcribed mRNA.

Embodiment 9mRNA-LNP entrapment

The mRNA stock solution was dispersed in 20 mM acetic acid solution (pH 5.0) to a final concentration of 200 μg/mL (aqueous phase). According to the example compound: cholesterol: DSPC: DMG-PEG2000 = 50: 38.5: 10: 1.5 molar ratio to mix into mixed fat (oil phase). The flow rate of the water phase and the oil phase is controlled to mix the mRNA and the lipid mixture at a volume ratio of 3:1 by means of T-mixed flow to obtain the mRNA entrapped by LNP. The loaded LNP was diluted 10 times with buffer, then concentrated by ultrafiltration, and the diluent was replaced, and finally the LNP was concentrated to mRNA to 200ug/mL, and the pH of LNP was adjusted to about 7-8. Finally, Ribogreen kit and 10% OTG were used as demulsifier to detect the total mRNA content and free content in LNP, and calculate the encapsulation efficiency of LNP. Dilute the LNP final product 10 times with the diluent, add 1ml into the particle size pool, put it on the Malvern ZetaSizer instrument, and detect the particle size of LNP. The results are shown in Table 1.

Table 1: Characterization data of the LNP of the example compounds

serial number particle size PDI encapsulation rate Example 1 72.68 0.0892 96 Example 2 72.02 0.0862 96 Example 3 71.55 0.0835 94 Example 4 70.42 0.0816 93 Example 5 70.67 0.0865 91 Example 6 70.88 0.0992 94 Example 7 72.73 0.0987 92 SM102 92.16 0.1296 95

The results show that the particle size of the LNP formed by the compound of the example is smaller than that of SM102, the uniformity is better than that of SM102, and the encapsulation efficiency of some compounds is better, indicating that the compound of the present invention has more ideal delivery ability and better product uniformity Sexuality and specific needs drug delivery.

Example 10 In vivo reporter gene expression detection in mice

The prepared mRNA solution entrapped by LNP is taken and diluted with PBS buffer solution to obtain a solution for injection. BALB/c female mice of about 20 g were injected with the injection solution at the quadriceps muscle of the mice with an insulin syringe, and each mouse was injected with 50 μl. Two doses were designed: one dose was "5 μg mRNA per 50 microliter injection", and the other was "14 μg mRNA per 50 microliter injection". About 20 g of BALB/c female mice were injected with PBS buffer solution in the quadriceps muscle of the mice with an insulin syringe, and each mouse was injected with 50 μl.

24h after mice were injected, the in vivo expression of Luciferase was detected by IVIS of Perkinelmer. The substrate is D-luciferin Sodium salt (GOLDBIO, LUCNA-1G), prepared with normal saline to a concentration of 15 mg/ml, sterilized by filtration with a 0.22 μm filter membrane, aliquoted and stored in the dark at -20°C. Before imaging, each mouse of about 20 g was injected intraperitoneally with 200 μl of substrate solution for 10-20 minutes, and then anesthetized with isoflurane gas, and then placed the mouse on the imaging plate to detect the fluorescence of the living animal.

The results showed that the LNP-transfected luciferase of lipid examples 1 and 2 had higher expression of luciferase in vivo than that of SM102 (see Table 2).

Table 2: In vivo expression levels of luciferase mRNA in mice delivered by cationic lipid LNP

The above results prove that the compounds 1 and 2 of Example can efficiently deliver mRNA in vivo, ensure the protein expression level, and the expression efficiency is higher than that of SM102.

Embodiment 11 lipid acute toxicity analysis

In order to examine the biological safety of the compounds of the examples, we analyzed the acute toxicity of compounds 1, 2 and 7. BALB/c female mice were enrolled in the experiment, with 10 mice in each group, and the high and low concentration groups were set at 50ug/mouse and 150ug LNP/mouse respectively. Take SM102 as the control. 24 hours after the mice were injected, blood was collected through the orbit, and about 100ul of blood was collected into a blood collection tube containing EDTA. After sampling, the blood was thoroughly mixed with the anticoagulant by inverting it up and down gently several times. Store at 4°C, do not directly contact with ice packs, and do not hit violently. The test results are shown in Table 3 and Table 4.

Table 3: Acute Toxicity Analysis of Cationic Lipid LNP (Low Dose Group)

Table 4: Acute Toxicity Analysis of Cationic Lipid LNP (High Dose Group)

The results showed that, compared with the control group, six major changes in creatinine, total bilirubin, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and γ-glutamyl transpeptidase in each group Toxicological indicators have no significant changes, which preliminarily shows the safety of this product. Compared with the SM102 group, the values of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and γ-glutamyl transpeptidase all decreased by 10-30%, indicating that the hepatic and renal toxicity of this product is further enhanced. decreased, indicating its higher biosafety.

Example 12 In vivo expression distribution analysis

In order to investigate the distribution characteristics of lipid-delivered mRNA in vivo, we analyzed the distribution characteristics of C57BL/6J mice given a single intramuscular injection of compound 1, 2 and 7 carrying reporter gene luciferase mRNA. Ten C57BL/6J mice were enrolled in the experiment, half male and half male, and 50 μg was administered to each mouse, with SM102 as the control.

The animals in the negative control group collected whole blood, bone marrow, liver, spleen, heart, kidney, Inguinal lymph nodes, mesenteric lymph nodes, spleen, brain, stomach, small intestine, non-injection site muscle, injection site muscle tissue, etc. The RT-PCR method was used to detect the RNA content in samples at each time point, and the quantitative lower limit of the method was 40copies/reaction to reflect the distribution characteristics in C57BL/6J mice.

The results showed that in SM102-injected animals, the exposure in tissue organs from high to low was as follows: inguinal lymph node, muscle at injection site, whole blood, muscle at non-injection site, spleen, bone marrow, heart, mesenteric lymph node, liver, kidney , small intestine, and brain; in the animals injected with lipid examples 1, 2, and 7, the exposure in tissue organs from high to low is as follows: muscle at the injection site, inguinal lymph nodes, whole blood, spleen, and non-injection sites Muscle, bone marrow, heart, liver, mesenteric lymph nodes, kidney, small intestine, and brain (see Figure 1). The research results show that the distribution of mRNA delivered by Examples 1 and 2 is higher than that of SM102 in circulation, immune system and muscle, but lower than that of SM102 in liver, heart and kidney. In addition, the distribution of mRNA delivered by the compound of Example 7 was higher in the stomach. This result suggests that, compared with SM102, the lipid obtained in Example 1 and Example 2 of this class of lipids is more suitable as a delivery carrier and is more suitable for products such as vaccines, and at the same time reduces the lipids in the liver, heart and kidney. Aggregation reduces its potential toxicity. The lipid obtained in Example 7 has certain advantages in gastric delivery.

Claims (9)

1. A long-chain alkyl ester amine compound, characterized in that, the structural formula is as follows: 2. A method for preparing the compound according to claim 1, comprising the steps of: Wherein, X is -(CH ₂ ) ₅ -; Step 1: Add OH-X-NH ₂ and DIEA to the protic solvent solution of compound A, heat and react for 12 to 36 hours, dilute with water, extract with ethyl acetate, and purify to obtain compound B; Step 2: Add compound B, potassium carbonate and potassium iodide to the solution of compound C in an aprotic solvent, heat and react for 24-48 hours, dilute with water, extract with ethyl acetate, and purify to obtain the target compound. 3. The method according to claim 2, wherein the protic solvent is selected from any one or a combination of ethanol and propanol. 4. The method according to claim 2, wherein the aprotic solvent is selected from any one or a combination of two or more of 2-methyltetrahydrofuran, cyclopentyl methyl ether or acetonitrile. 5. method according to claim 2, is characterized in that, described aprotic solvent is the mixed solvent of any one in 2-methyltetrahydrofuran or cyclopentyl methyl ether and acetonitrile, and described 2-methyltetrahydrofuran Or the ratio of any one of cyclopentyl methyl ether to acetonitrile is 1-3:1. 6. A composition, characterized in that, the composition comprises a therapeutic or preventive agent and a carrier for delivering the therapeutic or preventive agent, the carrier comprising the long-chain alkyl ester amines according to claim 1 compound. 7. The composition according to claim 6, wherein the therapeutic or preventive agent is selected from one or more of nucleic acid molecules, polypeptides, proteins or small molecule compounds. 8. The application of the compound according to claim 1 in the preparation of nucleic acid drugs, vaccines, protein or polypeptide drugs, and small molecule drugs. 9. The application of the compound according to claim 1 in the preparation of mRNA medicine or vaccine.