Guanidyl-derived ionizable lipid material, preparation method and application

Abstract

The invention provides a guanidyl-derived ionizable lipid material. The guanidyl-derived ionizable lipid material is a compound as shown in a formula (I), and a solvent crystal substance or an isomeride or a pharmaceutically acceptable salt of the compound as shown in the formula (I), in the imgabs0 #, R1 is a substituted or non-substituted linear alkyl alcohol of C2-C6, and R2 is a substituted or non-substituted linear alkyl alcohol of C2-C6; r < 2 >, R < 3 >, R < 4 > and R < 5 > are all C6-C18 substituted or unsubstituted alkyl, alkenyl or alkynyl, and comprise or do not comprise one or more heteroatoms; r < 2 >, R < 3 >, R < 4 > and R < 5 > are the same or different; n is a positive integer from 3 to 18; and m is a positive integer from 3 to 18. The guanidyl-derived ionizable lipid material disclosed by the invention has excellent biocompatibility, efficient drug delivery performance, efficient transfection efficiency, wide application potential and simplicity and convenience in preparation and can be produced on a large scale, and the stability, targeting and drug release control of an inner body membrane are effectively destroyed.

Description

Technical Field

[0001] The invention belongs to the field of biomedical carriers, and in particular relates to a guanidine-derived ionizable lipid material, a preparation method and an application thereof. Background Art

[0002] As an emerging treatment strategy, gene therapy has shown good application prospects for pathological conditions related to abnormal gene expression, such as cancer, vitiligo, scars, and ophthalmic diseases. However, the special structure of nucleic acid gene drugs (such as small interfering RNA, messenger RNA, and plasmid DNA) makes them susceptible to nuclease degradation in the body, thereby reducing their bioavailability and therapeutic effects. In addition, the delivery efficiency of nucleic acid gene drugs is often limited by the ability to penetrate the cell membrane, resulting in the inability of the drug to effectively reach the target cells; therefore, the effectiveness of nucleic acid gene drugs depends on efficient and targeted delivery vectors to ensure that the drug can cross the cell membrane and successfully reach the target cell.

[0003] In recent years, biomedical carrier materials have shown significant potential in the delivery of nucleic acid-based gene drugs. Lipid nanoparticles (LNPs) have attracted widespread attention due to their good biocompatibility, stability and certain drug delivery capabilities. However, current LNP carriers still face multiple challenges in gene drug delivery, including low delivery efficiency, insufficient targeting and rapid degradation in the body.

[0004] In the study of LNP carriers, guanidine-derived ionizable lipid materials show good drug delivery potential due to their unique chemical structure and physicochemical properties. Guanidine compounds have strong alkalinity and can form stable complexes with biological molecules such as nucleic acids, thereby effectively loading nucleic acid gene drugs. In addition, drug-loaded LNPs prepared from this ionizable lipid material have excellent drug release characteristics in an acidic environment, which is suitable for targeted release in the tumor microenvironment, thereby enhancing the therapeutic effect.

[0005] Based on this, developing a new type of guanidine-derived ionizable lipid material as the core material of LNP carriers in order to improve the delivery efficiency of nucleic acid gene drugs and enhance their stability and targeting in vivo is a technical problem that needs to be urgently solved in this field. Summary of the invention

[0006] In view of the deficiencies in the prior art, the object of the present invention is to provide a guanidine-derived ionizable lipid material, a preparation method and an application thereof.

[0007] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a guanidine-derived ionizable lipid material, which is a compound represented by formula (I);

[0009]

[0010] Wherein, R1 is a C2-C6 substituted or unsubstituted straight-chain alkyl alcohol;

[0011] R2, R3, R4 and R5 are all C6-C 18 A substituted or unsubstituted alkyl, alkenyl or alkynyl group, including or excluding one or more heteroatoms; said R2, said R3, said R4, said R5 are the same or different from each other;

[0012] n is a positive integer from 3 to 18;

[0013] m is a positive integer from 3 to 18.

[0014] Furthermore, the guanidine-derived ionizable lipid material provided by the present invention may also be a solvent crystal, an isomer or a pharmaceutically acceptable salt of the compound of the general formula (I).

[0015] Furthermore, the guanidine-derived ionizable lipid material provided by the present invention is one of P6a, P10a, P12a, P6b, P10b, P12b, P6c, P10c, and P12c, and its structural formula is as follows:

[0016]

[0017]

[0018] In a second aspect, the present invention further provides a method for preparing a guanidine-derived ionizable lipid material, comprising:

[0019] In an anhydrous solvent, in the presence of a catalyst, a carboxylic acid derivative and a guanidine derivative are used as raw materials and are obtained by covalent bond coupling. The carboxylic acid derivative is represented by formula (II), and the guanidine derivative is represented by formula (III):

[0020]

[0021] Wherein, in formula (II) and (III), R1, R2, R3, and n have the same meanings as in the compound of formula (I);

[0022] X1 is bromine or chlorine.

[0023] Further, the specific steps are as follows:

[0024] S1. covalently coupling the carboxylic acid derivative and the guanidine derivative using anhydrous dichloromethane as solvent;

[0025] S2. The coupling reaction catalyst in step S1 is a combination of one or more of dicyclohexylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, diisopropylcarbodiimide, 4-N,N-lutidine, and 1-hydroxybenzotriazole;

[0026] S3. The reaction temperature is room temperature and the reaction time is 6-12h.

[0027] Furthermore, the preparation method of the carboxylic acid derivative is as follows:

[0028] In anhydrous dichloromethane, bromohydrin b and 2-hexylundecanoic acid are used as raw materials and esterified under the catalysis of EDCI / DMAP to obtain;

[0029] Furthermore, the preparation method of the guanidine derivative is as follows:

[0030] i. In anhydrous dichloromethane, under the catalysis of acetic anhydride and triethylamine, 3-amino-1-propanol and Boc-S-methylisothiourea hemisulfate are reacted as raw materials to obtain intermediate 3; the intermediate 3 is represented by formula (IV):

[0031]

[0032] ii. Under the action of trifluoroacetic acid, the intermediate 3 is deprotected to obtain the intermediate 4; the intermediate 4 is represented by formula (V):

[0033]

[0034] iii. In anhydrous tetrahydrofuran, under the catalysis of sodium hydride, the intermediate 4 undergoes a nucleophilic substitution reaction with a halohydrin to obtain the guanidine derivative.

[0035] In a third aspect, the present invention also provides the use of the guanidine-derived ionizable lipid material for preparing lipid nanoparticles (LNP) for delivering nucleic acid gene drugs.

[0036] In a fourth aspect, the present invention provides a lipid nanoparticle (LNP) composition, comprising the guanidine-derived ionizable lipid material. The lipid nanoparticle composition may further comprise auxiliary lipids, cholesterol or its derivatives, polyethylene glycol lipids and nucleic acid gene drugs.

[0037] Furthermore, the auxiliary lipid is selected from a combination of one or more of dioleoylphosphatidylethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphocholine, 1,2-docosayl-sn-glycero-3-phosphocholine, 1,2-eicosanoyl-sn-glycero-3-phosphocholine, aminophosphatidic acid, hexadecyl lipid, and dioctadecyl lipid; preferably DOPE.

[0038] Furthermore, the cholesterol derivative is selected from one or more combinations of cholesterol-PEG, cholesterol phosphate, cholesterol-amino acid, cholesterol-fatty acid, and cholesterol-dimer.

[0039] Furthermore, the polyethylene glycol lipid is selected from a combination of one or more of distearoylphosphatidylcholine-polyethylene glycol (DSPC-PEG), eicosylphosphatidic acid polyethylene glycol, eicosylphosphatidic acid polyethylene glycol, polyethylene glycol octadecyl (C18-PEG), and tetracosylphosphatidic acid polyethylene glycol; preferably DSPC-PEG.

[0040] Furthermore, the nucleic acid gene drug includes siRNA, mRNA, plasmid DNA, circular RNA, long non-coding RNA, antisense oligonucleotides, small activating RNA, nucleic acid aptamers or a combination of one or more sequences targeting specific targets; most preferably, the nucleic acid gene drug is mRNA.

[0041] Furthermore, the molar ratio of the guanidine-derived ionizable lipid material, the auxiliary lipid, the cholesterol or its derivative, and the polyethylene glycol lipid is 30-50:20-30:5-10:20-30; the molar ratio of the guanidine-derived ionizable lipid material to the nucleic acid gene drug is 3-5:1.

[0042] In a fifth aspect, the present invention provides a method for preparing the lipid nanoparticles (LNP), including nanoprecipitation method, thin film hydration method, injection method, ultrasonic method, microfluidics method, solvent volatilization method, reverse phase evaporation method, and electrospray method.

[0043] Preferably, a microfluidic method is used, and the specific steps of the preparation method include:

[0044] i. dissolving the guanidine-derivatized ionizable lipid material, the auxiliary lipid, the cholesterol or its derivative, and the polyethylene glycol lipid in ethanol to obtain an ethanol phase solution;

[0045] ii. dissolving the nucleic acid gene drug in 10-50 mM citric acid buffer (pH=4) to form an aqueous solution;

[0046] iii. Using a microfluidic mixer, the ethanol phase solution and the aqueous phase solution are mixed in a ratio of 1:3, dialyzed, and filtered to obtain the LNP.

[0047] The guanidine-derived ionizable lipid material provided by the present invention not only optimizes the design of lipid nanoparticles (LNPs), but also enhances the delivery efficiency and therapeutic effect of nucleic acid gene drugs; the specific advantages are as follows:

[0048] 1. Excellent biocompatibility

[0049] The guanidine-derived ionizable lipid material of the present invention has good biocompatibility, reduces toxicity and immune response in the body, and ensures that adverse reactions are reduced during the treatment process; such biocompatibility makes it suitable for various clinical applications, especially for long-term and high-dose treatment;

[0050] 2. Efficient drug delivery performance

[0051] The guanidine-derived ionizable lipid material of the present invention has good nucleophilicity and positive charge ability, and can effectively combine with negatively charged nucleic acid gene drugs (such as siRNA, mRNA, etc.) to form stable nanoparticles, thereby significantly improving the drug delivery efficiency; in addition, under physiological conditions, this carrier can regulate the surface charge to make LNP circulate in the body for a longer time and enhance the targeting effect;

[0052] 3. High transfection efficiency

[0053] The guanidine-derived ionizable lipid material design of the present invention can ensure efficient cell transfection; the lipid material exhibits a positive charge in an acidic environment (such as in endosomes), can interact with anionic intracellular lipids, and enhance cell membrane permeability, thereby effectively delivering nucleic acid gene drugs into cells; this mechanism improves the transfection efficiency of gene drugs, thereby enhancing the therapeutic effect;

[0054] 4. Effectively destroy the stability of endosomal membrane

[0055] The guanidine-derived ionizable lipid material of the present invention can form a positive charge in an acidified endosomal environment (pH≈5-6), and promote the formation of a hexagonal (HII) phase through its unique cone structure; this structure helps to destroy the stability of the endosomal membrane, thereby achieving the effective release of nucleic acid gene drugs, overcoming the bottleneck problem of traditional carriers in nucleic acid release, and significantly improving the drug delivery efficiency;

[0056] 5. Targeting and drug release control

[0057] By designing the acid dissociation constant (pKa) of the guanidine-derived ionizable lipid material of the present invention, the present invention can ensure that the carrier maintains a neutral charge at physiological pH, reduce nonspecific binding, reduce side effects, and trigger positive charges through an acidic microenvironment (such as a tumor or diseased tissue) to promote targeted release of the drug. This property has significant advantages for the treatment of tumors and other diseases related to acidic microenvironments;

[0058] 6. Wide application potential

[0059] The guanidine-derived ionizable lipid material of the present invention can be used not only for cancer treatment, but also for other diseases associated with abnormal gene expression, including vitiligo, scars and ophthalmic diseases; due to its excellent nucleic acid delivery ability, the technology platform can be adapted to a variety of different types of gene drugs, such as mRNA vaccines, siRNA therapies and DNA vaccines;

[0060] 7. Easy preparation and scalable production

[0061] The synthesis method of the guanidine-derived ionizable lipid material of the present invention is relatively simple, can be obtained through a conventional chemical synthesis route, and is suitable for large-scale production; the preparation process has high controllability, ensures the stability and consistency of production, and is suitable for industrial application. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] In order to more clearly illustrate the technical solutions in the present invention, the following provides figures in various embodiments, and all figures are limited to the embodiments in the present invention.

[0063] Figure 1 A synthetic route diagram of a guanidine-derived ionizable lipid material of the present invention;

[0064] Figure 2 Characterization of the particle size and PDI of the lipid nanoparticles in Example 12 of the present invention;

[0065] Figure 3 is the surface potential of the lipid nanoparticles in Example 12 of the present invention;

[0066] Figure 4 Characterization of particle size and PDI of lipid nanoparticles in PBS and plasma in Example 12 of the present invention;

[0067] Figure 5 Zeta potential characterization of lipid nanoparticles in PBS and plasma in Example 12 of the present invention;

[0068] Figure 6 is a transmission electron microscopy image of the lipid nanoparticles in Example 12 of the present invention;

[0069] Figure 7 The results of the determination of the Fluc-eGFP mRNA encapsulation efficiency of lipid nanoparticles in Example 12 of the present invention;

[0070] Figure 8 This is the result of investigating the transfection efficiency of Fluc-eGFP mRNA of lipid nanoparticles in Example 12 of the present invention;

[0071] Fig. 9 The results of fluorescence microscopy observation of 293T cells transfected with Fluc-eGFP mRNA of lipid nanoparticles in Example 12 of the present invention. DETAILED DESCRIPTION

[0072] In view of the deficiencies in the prior art, the object of the present invention is to provide a guanidine-derived ionizable lipid material and a preparation method and application thereof.

[0073] The technical solution provided in this application is synthesized by independent design to obtain guanidine-derived ionizable lipid materials, and by screening the ratio of auxiliary lipids, cholesterol or its derivatives, and polyethylene glycol lipids, lipid nanoparticles (LNP) are prepared by microfluidic technology for the effective delivery of nucleic acid gene drugs. It is verified at the cellular level that it has efficient transfection ability, can efficiently deliver nucleic acid gene drugs, and significantly improve the transfection efficiency and targeting of gene drugs. The lipid has excellent biocompatibility and drug delivery performance under physiological conditions, can destroy the stability of endosomal membranes in acidic microenvironments, and realize the effective release of nucleic acids, thereby enhancing the effect of gene therapy. The invention is applicable to a variety of clinical treatment fields such as cancer, vitiligo, scars and ophthalmic diseases, and has broad application prospects.

[0074] In order to make the present application easier for readers to understand, the following is a detailed description in the form of examples; the intermediates mentioned in the examples have chemical formulas in the attached Figure 1 Those skilled in the art should understand that the embodiments are only used to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0075] The instruments and reagents used in the examples are as follows:

[0076] 1. Instrument

[0077] Nuclear magnetic resonance spectrometer, magnetic stirrer, centrifuge, Malvern laser particle size analyzer, fluorescence microscope, laser confocal microscope, freeze dryer, water purifier, transmission electron microscope.

[0078] 2. Reagents

[0079] Fluc-eGFP mRNA, PBS, Australian fetal Gibco10099141 serum, DMEM medium, Quant-iTRiboGreen RNA Assay kit, 293T cells.

[0080] Embodiment 1:

[0081] Synthesis of intermediate b:

[0082] 6-Bromo-1-hexanol (1.8 g, 10 mmol) was dissolved in dichloromethane, 2-hexyl undecanoic acid (2.56 g, 10 mmol) and EDCI / DMAP were added, the reaction was carried out at room temperature for 6 h, and intermediate b (3.137 g, yield was 75.1%) was obtained by recrystallization.

[0083] Embodiment 2:

[0084] Synthesis of intermediate 4:

[0085] Boc-S-methylisothiourea sulfate (1.45 g, 5 mmol, 1 eq) was dissolved in DMF (10 mL), 3-amino-1-propanol (380 mg, 5 mmol, 1 eq) was added at 25 ° C. After reacting for 9 h, the mixture was washed with water and extracted with DCM. The lower layer of the solution was collected, evaporated to dryness, and column chromatography (petroleum ether: ethyl acetate = 2:1) was performed to obtain an amorphous white solid 2 (1.24 g, yield 90%); compound 2 (0.5 g) was dissolved in 5 mL of dichloromethane, 200 μL of acetic anhydride and 450 μL of triethylamine were added, mixed evenly, and stirred at room temperature for 12 h. After the reaction was completed, saturated ammonium chloride solution was added for washing, brine was added for washing, and compound 3 was evaporated to dryness; compound 3 (0.5 g) was dissolved in an appropriate amount of dichloromethane, trifluoroacetic acid was added, and the reaction was stirred at room temperature for 3 h. After drying, intermediate 4 (217.36 mg, 98%) was obtained.

[0086] Embodiment 3:

[0087] Synthesis of key intermediate P6a-1:

[0088] In anhydrous tetrahydrofuran, under the catalysis of sodium hydride, intermediate b (418 mg, 1 mmol) and intermediate 4 (79.5 mg, 0.5 mmol) were covalently coupled at a molar ratio of 2:1 to obtain intermediate P6a-1 (263.36 mg, yield 63%).

[0089] Embodiment 4:

[0090] Synthesis of final product P6a:

[0091] P6a-1 (835 mg, 1 mmol) was dissolved in sodium methoxide solution (5 mL, 1 N) in methanol and stirred at room temperature for 3 h. Dilute hydrochloric acid was added for washing, and dichloromethane was used for extraction. The organic solvent was evaporated to dryness to obtain P6a (702 mg, yield: 89.12%).

[0092] Example 5

[0093] The synthesis of intermediates d and f was as described in Example 1, except that 6-bromo-1-hexanol was replaced by 10-bromo-1-decanol and 12-bromo-1-dodecanol (10 mmol), respectively; the other steps and conditions were the same as in Example 1.

[0094] Example 6

[0095] The synthesis of intermediates 8 and 12 was as described in Example 2, except that 3-amino-1-propanol was replaced by 4-amino-1-butanol and 5-amino-1-pentanol (5 mmol), respectively; the other steps and conditions were the same as in Example 2.

[0096] Example 7

[0097] The synthesis of key intermediate P10a-1 and key intermediate P12a-1 is as described in Example 3, except that intermediate b is replaced by intermediate d and f, respectively, and the other steps and conditions are consistent with Example 3.

[0098] Example 8

[0099] The synthesis of key intermediate P6b-1 and key intermediate P6c-1 is as described in Example 3, except that intermediate 4 is replaced by intermediates 8 and 12, respectively, and the other steps and conditions are consistent with Example 3.

[0100] Example 9

[0101] The synthesis of key intermediates P10b-1 and key intermediates P12b-1 is as described in Example 3, except that intermediate 4 is replaced by intermediate 8, and intermediate b is replaced by intermediates d and f, respectively. The other steps and conditions are consistent with Example 3.

[0102] Example 10

[0103] The synthesis of key intermediate P10c-1 and key intermediate P12c-1 is as described in Example 3, except that intermediate 4 is replaced by intermediate 12, and intermediate b is replaced by intermediates d and f, respectively. The other steps and conditions are consistent with Example 3.

[0104] Embodiment 11

[0105] The synthesis of the final products P6b, P6c, P10a, P10b, P10c, P12a, P12b, and P12c is as described in Example 4, except that the key intermediate P6a-1 is replaced by P6b-1, P6c-1, P10a-1, P10b-1, P10c-1, P12a-1, P12b-1, and P12c-1, and the other steps and conditions are the same as in Example 4.

[0106] Example 12

[0107] The guanidine-derived ionizable lipid material P10b was selected as a representative compound to prepare lipid nanoparticles (LNP). The guanidine-derived ionizable lipid material (P10b), DOPE, cholesterol and DSPC-PEG50:20:10:20 molar ratio were dissolved in ethanol to prepare an ethanol phase solution. Next, Fluc-eGFP mRNA was added to 50mM citrate buffer (pH=4) to form an aqueous phase solution. Lipid nanoparticles (LNP) were obtained by uniformly mixing the ethanol phase solution with the aqueous phase solution (1:3) using a microfluidic mixer. After filtration, all samples were stored at 4°C. Among them, the molar ratio of guanidine-derived ionizable lipid material (P10b) to Fluc-eGFP mRNA was 5:1. According to the above operation and prescription ratio, LNP was prepared and labeled as P10b LNP for further characterization.

[0108] Embodiment 13

[0109] Characterization of Guanidine-derivatized Ionizable Lipid Nanoparticles:

[0110] The particle size, Zeta potential, and polydispersity index (PDI) of the lipid nanoparticles (LNP) prepared in Example 12 were measured using a dynamic light scattering laser particle size analyzer (DLS). The Quant-iT RiboGreen RNA Assay kit was used to determine the Fluc-eGFP mRNA encapsulation efficiency in the LNP. The average particle size of the LNP was measured to be 85 nm, the particle size distribution was relatively uniform, and the polydispersity index (PDI) was 0.18, indicating that the LNP had good uniformity and a potential close to neutral.

[0111] Embodiment 14

[0112] The in vitro transfection efficiency of P10b LNP was investigated, and the specific method included the following steps:

[0113] (1) The negative control group was set up with Fluc-eGFP mRNA basal culture medium solution, and the positive control was set up with DLin-MC3

[0114] LNP (encapsulated with Fluc-eGFP mRNA) basal culture medium solution, and the drug-treated group received the basal culture medium solution of P10b LNP;

[0115] (2) 293T cells in the logarithmic growth phase were seeded in 6-well plates at 200,000 cells per well and incubated for 12 h before transfection;

[0116] (3) The negative control group was added with the basal culture medium solution of Fluc-eGFP mRNA, and the drug-treated group was added with the basal culture medium solution of P10b LNP so that each well contained 0.2 μg of Fluc-eGFP mRNA;

[0117] (4) 24 h after transfection, flow cytometry analysis was performed, and the obtained fluorescence data were processed using FlowJo 10 software.

[0118] The overall experimental results show that the nucleic acid delivery efficiency of guanidine-derived ionizable lipid materials with different structures is significantly different ( Figure 8 Only LNP prepared by P10b is shown, and different LNP preparation processes will affect the particle size and polydispersity coefficient of LNP, but they are basically kept within 200. The flow cytometry results show that compared with LNP prepared by commercially available DLin-MC3, the guanidine-derived ionizable lipid material P10b provided by the present invention has the same excellent nucleic acid gene drug delivery performance, can efficiently load and release nucleic acid gene drugs, improves the transfection efficiency of nucleic acid gene drugs, effectively overcomes the limitations of existing delivery systems in clinical applications, greatly enhances the effect of gene therapy, and has broad application prospects.

[0119] The applicant declares that the present invention illustrates the preparation method and application of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned method steps, that is, it does not mean that the present invention must rely on the above-mentioned preparation method steps to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials selected by the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A guanidine-derived ionizable lipid material, characterized in that: It is a compound represented by formula (I) or a solvent crystal, isomer, or pharmaceutically acceptable salt of the compound: Wherein, R1 is a C2-C6 substituted or unsubstituted straight-chain alkyl alcohol; R2, R3, R4 and R5 are all C6-C 18 A substituted or unsubstituted alkyl, alkenyl or alkynyl group, including or excluding one or more heteroatoms; said R2, said R3, said R4, said R5 are the same or different from each other; n is a positive integer from 3 to 18; m is a positive integer from 3 to 18.

2. The guanidine-derived ionizable lipid material according to claim 1, characterized in that It is one of P6a, P10a, P12a, P6b, P10b, P12b, P6c, P10c, P12c, and the corresponding structural formula is as follows: 。 3. A method for preparing a guanidine-derived ionizable lipid material, characterized in that: include: In an anhydrous solvent, in the presence of a catalyst, a carboxylic acid derivative and a guanidine derivative are used as raw materials and are obtained by covalent bond coupling. The carboxylic acid derivative is represented by formula (II), and the guanidine derivative is represented by formula (III): Wherein, R1 is a C2-C6 substituted or unsubstituted straight-chain alkyl alcohol; R2 and R3 are both C6-C 18 A substituted or unsubstituted alkyl, alkenyl or alkynyl group, including or excluding one or more heteroatoms; said R2 and said R3 are the same or different from each other; n is a positive integer from 3 to 18; X1 is bromine or chlorine.

4. The method for preparing the guanidine-derived ionizable lipid material according to claim 3, characterized in that: The specific steps are as follows: S1. covalently coupling the carboxylic acid derivative and the guanidine derivative using anhydrous dichloromethane as solvent; S2. The coupling reaction catalyst in step S1 is a combination of one or more of dicyclohexylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, diisopropylcarbodiimide, 4-N,N-lutidine, and 1-hydroxybenzotriazole; S3. The reaction temperature is room temperature and the reaction time is 6-12h.

5. The method for preparing the guanidine-derived ionizable lipid material according to claim 4, characterized in that: The preparation method of the carboxylic acid derivative is as follows: In anhydrous dichloromethane, bromohydrin b and 2-hexylundecanoic acid are used as raw materials and esterified under the catalysis of EDCI / DMAP to obtain; Or, the preparation method of the guanidine derivative is as follows: i. In anhydrous dichloromethane, under the catalysis of acetic anhydride and triethylamine, 3-amino-1-propanol and Boc-S-methylisothiourea hemisulfate are reacted as raw materials to obtain intermediate 3; the intermediate 3 is represented by formula (IV): ii. Under the action of trifluoroacetic acid, the intermediate 3 is deprotected to obtain the intermediate 4; the intermediate 4 is represented by formula (V): iii. In anhydrous tetrahydrofuran, under the catalysis of sodium hydride, the intermediate 4 undergoes a nucleophilic substitution reaction with a halohydrin to obtain the guanidine derivative.

6. Use of the guanidine-derived ionizable lipid material as described in any one of claims 1 or 2 for preparing lipid nanoparticles (LNP) for delivering nucleic acid gene drugs.

7. A lipid nanoparticle composition, characterized in that: The invention comprises the guanidine-derived ionizable lipid material as described in any one of claims 1 or 2, auxiliary lipids, cholesterol or its derivatives, polyethylene glycol lipids and nucleic acid gene drugs.

8. The lipid nanoparticle composition according to claim 7, wherein The auxiliary lipid is a combination of one or more of dioleoylphosphatidylethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphocholine, 1,2-docosayl-sn-glycero-3-phosphocholine, 1,2-eicosanoyl-sn-glycero-3-phosphocholine, aminophosphatidic acid, hexadecyl lipid, and dioctadecyl lipid; Or, the cholesterol derivative is a combination of one or more of cholesterol-PEG, cholesterol phosphate, cholesterol-amino acid, cholesterol-fatty acid, and cholesterol-dimer; Or, the polyethylene glycol lipid is a combination of one or more of distearoylphosphatidylcholine-polyethylene glycol (DSPC-PEG), eicosylphosphatidic acid polyethylene glycol, eicosylphosphatidic acid polyethylene glycol, polyethylene glycol octadecyl (C18-PEG), and tetracosylphosphatidic acid polyethylene glycol; Or, the nucleic acid gene drug is a combination of one or more of siRNA, mRNA, plasmid DNA, circular RNA, long non-coding RNA, antisense oligonucleotide, small activating RNA, nucleic acid aptamer or sequences targeting specific targets; Or, the molar ratio of the guanidine-derived ionizable lipid material, the auxiliary lipid, the cholesterol or its derivative, and the polyethylene glycol lipid is 30-50:20-30:5-10:20-30; the molar ratio of the guanidine-derived ionizable lipid material to the nucleic acid gene drug is 3-5:

1.

9. The method for preparing lipid nanoparticles according to any one of claims 7 or 8, characterized in that: Including nanoprecipitation method, thin film hydration method, injection method, ultrasonic method, microfluidics method, solvent volatilization method, reverse phase evaporation method, and electrospray method.

10. The method for preparing lipid nanoparticles according to claim 9, characterized in that: The specific steps of the preparation method using the microfluidic method include: i. dissolving the guanidine-derivatized ionizable lipid material, the auxiliary lipid, the cholesterol or its derivative, and the polyethylene glycol lipid in ethanol to obtain an ethanol phase solution; ii. dissolving the nucleic acid gene drug in 10-50 mM citric acid buffer (pH=4) to form an aqueous solution; iii. using a microfluidic mixer to mix the ethanol phase solution with the aqueous phase solution at a ratio of 1:3, The lipid nanoparticles are obtained by filtration.

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