Lipid nanoparticle, composition and application thereof

Abstract

The invention belongs to the technical field of biology, particularly relates to a lipid nanoparticle, a composition and application of the lipid nanoparticle, and more particularly relates to a lipid nanoparticle composed of ganglioside glycolipid, a composition and application of the lipid nanoparticle. The invention provides a lipid nanoparticle. The lipid nanoparticle comprises ganglioside glycolipid, cationic lipid, auxiliary phospholipid and steroid lipid. The lipid nanoparticles provided by the invention are better in encapsulation efficiency, good in effect during repeated administration, and better in-vivo delivery efficiency.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and in particular, relates to a lipid nanoparticle, a composition and application thereof, and more particularly, to a lipid nanoparticle composed of ganglioside lipids, a composition and application thereof. BACKGROUND

[0002] mRNA vaccines and drugs topped the list of the top ten breakthrough technologies in the world in 2021 for their great changes in the medical field. However, mRNA is a high-molecular-weight negatively charged molecule with extremely strong hydrophilicity, and it is difficult to penetrate the cell membrane into the cell. In addition, mRNA is very unstable and easily degraded by nucleases, and has a very short circulation time in the blood. Therefore, it is necessary to develop a carrier and composition for promoting the in vivo delivery of mRNA.

[0003] Lipid nanoparticle (LNP) technology has high delivery efficiency and good safety, and is the most suitable carrier for delivering mRNA. LNP is usually composed of cationic lipids, auxiliary phospholipids, steroid lipids and polyethylene glycol (PEG) lipids. Among them, PEG lipids can stabilize LNP and increase its half-life in the body. However, due to the immunogenicity of polyethylene glycol to cells, some people have developed antibodies against polyethylene glycol in the body, which will cause the active ingredients to be quickly cleared by the immune system when the nucleic acid therapeutic agent is repeatedly injected, reducing the treatment or prevention effect. Since mRNA expression is transient, repeated administration is necessary to achieve the best therapeutic effect. Therefore, there is an urgent need to introduce new lipids with better properties to replace PEG lipids to solve the problem of poor effect of PEG-modified lipid nanoparticles when repeatedly administered.

[0004] There is a need in the art for a new type of lipid nanoparticle that can solve the problem of poor effect of PEG-modified lipid nanoparticles when repeatedly administered, while also having good in vivo biological effects. SUMMARY

[0005] Therefore, in a first aspect, the present application provides a lipid nanoparticle comprising a ganglioside lipid, a cationic lipid, an auxiliary phospholipid and a steroid lipid.

[0006] The use of the lipid nanoparticle of the present application has better encapsulation efficiency, good effect when repeatedly administered, and better delivery efficiency in vivo.

[0007] In some specific embodiments, the ganglioside lipid is a compound of formula (I) or formula (II) or a pharmaceutically acceptable salt thereof:

[0008]

[0009] wherein,

[0010] R 1 is selected from the group consisting of monosialyl fucosetetrahexose (FucGM1), monosialyl tetrahexose (GM1), monosialyl trihexose (GM2), monosialyl dihexose (GM3), disialyl dihexose (GD3), disialyl trihexose (GD2), disialyl tetrahexose (GD1a), disialyl tetrahexose (GD1b), or trisialyl dihexose (GT3);

[0011] R 2 is selected from the group consisting of substituted or unsubstituted linear or branched alkanes comprising 7 to 24 carbon atoms.

[0012] Further, at least one carbon atom of said R 2 is optionally replaced by at least one selected from the group consisting of -C(=O)-, -NH-, -O-, -S-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)NH-, or -NHC(=O)-.

[0013] Further, at least one carbon atom of said R 1 is selected from at least one of the following structures:

[0014]

[0015] Further, at least one carbon atom of said R 2 is selected from at least one of the following structures:

[0016]

[0017] In some specific embodiments, said ganglioside is selected from the group consisting of compounds represented by the following structures or a pharmaceutically acceptable salt thereof:

[0018]

[0019] Further, said ganglioside is selected from the group consisting of compounds represented by the following structure (I) or a pharmaceutically acceptable salt thereof:

[0020]

[0021] R 1 is monosialyl fucosetetrahexose;

[0022] R 2 is a linear alkyl group comprising 10 to 20 carbon atoms.

[0023] In some specific embodiments, said ganglioside is selected from the group consisting of compounds represented by the following structures or a pharmaceutically acceptable salt thereof:

[0024] In some specific embodiments, said ganglioside is selected from the group consisting of compounds represented by the following structures or a pharmaceutically acceptable salt thereof:

[0025] Specifically, R of FucGM1CerC14 is 1 a monosialylated fucosyl tetrahexose, R 2 a linear alkyl group C containing 13 carbon atoms 13 H 27 .

[0026] Specifically, R of FucGM1CerC18 is 1 a monosialylated fucosyl tetrahexose, R 2 a linear alkyl group C containing 17 carbon atoms 17 H 35 .

[0027] The lipid nanoparticles prepared using the above ganglioside lipids can obtain better in vivo biological effects, for example, the encapsulation efficiency and delivery efficiency thereof are better.

[0028] In some specific embodiments, the cationic lipid comprises one or more of 1-octyl nonyl 8-[(2-hydroxyethyl)[6-O-6-(undecyloxy)hexyl]amino]-octanoate (SM102) or 4-(N,N-dimethylamino)butyric acid (dilinoleyl) methyl ester (MC3).

[0029] In some specific embodiments, the helper phospholipid comprises one or more of phosphatidylcholine, phosphatidylethanolamine. Illustratively, the helper phospholipid comprises one or more of distearoylphosphatidylcholine (DSPC) or dioleoylphosphatidylethanolamine (DOPE).

[0030] In some specific embodiments, the sterol lipid comprises at least one of cholesterol, β-sitosterol, ergosterol, campesterol, stigmasterol, a corticosteroid. Illustratively, the sterol lipid comprises cholesterol or β-sitosterol.

[0031] In some specific embodiments, the molar ratio between the cationic lipid, the helper phospholipid, the sterol lipid, the ganglioside lipid is (30-60):(5-25):(25-50):(0.05-10).

[0032] In one specific embodiment, the molar ratio between the cationic lipid, the helper phospholipid, the sterol lipid, the ganglioside lipid is 50:10:38.5:1.5.

[0033] The lipid nanoparticles prepared using the above ratio have better biological effects.

[0034] In a second aspect, the present application provides a composition comprising the lipid nanoparticle as described above, wherein the lipid nanoparticle is loaded with an active agent.

[0035] In some embodiments, the active agent comprises at least one of a prophylactic agent, a therapeutic agent.

[0036] In some embodiments, the active agent comprises a nucleic acid, an immunomodulator, an antigen or fragment thereof, a vaccine, an anti-inflammatory agent, an anti-tumor agent, a small molecule drug, or a mixture thereof.

[0037] Further, the nucleic acid comprises at least one of a messenger RNA (mRNA), a small interfering RNA (siRNA), a microRNA, a single guide RNA (sgRNA), an antisense oligonucleotide (ASO), a DNA, a plasmid.

[0038] Further, the mRNA comprises a tumor antigen mRNA.

[0039] In a third aspect, the present application provides use of the lipid nanoparticle or the composition as described above in the manufacture of a medicament.

[0040] Further, the lipid nanoparticle or the composition can deliver the active agent in vivo.

[0041] In some embodiments, the active agent comprises at least one of a prophylactic agent, a therapeutic agent.

[0042] In some embodiments, the active agent comprises a nucleic acid, an immunomodulator, an antigen or fragment thereof, a vaccine, an anti-inflammatory agent, an anti-tumor agent, a small molecule drug, or a mixture thereof.

[0043] Further, the nucleic acid comprises at least one of a messenger RNA (mRNA), a small interfering RNA (siRNA), a microRNA, a single guide RNA (sgRNA), an antisense oligonucleotide (ASO), a DNA, a plasmid.

[0044] Further, the mRNA comprises a tumor antigen mRNA.

[0045] Further, the medicament can be used for cancer treatment and / or prevention. Still further, the cancer is melanoma.

[0046] Further, the medicament can be used for delivery to the spleen.

[0047] In a fourth aspect, the present application provides a method for preparing a lipid nanoparticle, comprising:

[0048] ganglioside, a cationic lipid, a helper phospholipid, and a sterol lipid are provided;

[0049] mixing and dissolving the above substances to obtain a mixture; and

[0050] processing the mixture to obtain a lipid nanoparticle.

[0051] Further, the preparation method further comprises a step of mixing the mixture with an active substance.

[0052] Further, the mixing ratio of the mixture to the active substance is (10:1) to (1:10).

[0053] In a specific embodiment, the mixing ratio of the mixture to the active substance is 1:3.

[0054] In some specific embodiments, the molar ratio of the cationic lipid, the helper phospholipid, the sterol lipid, and the ganglioside mixed together is (30-60):(5-25):(25-50):(0.05-10).

[0055] In a specific embodiment, the molar ratio of the cationic lipid, the helper phospholipid, the sterol lipid, and the ganglioside mixed together is 50:10:38.5:1.5.

[0056] In some specific embodiments, the active agent comprises at least one of a prophylactic agent, a therapeutic agent.

[0057] In some specific embodiments, the active agent comprises a nucleic acid, an immunomodulator, an antigen or a fragment thereof, a vaccine, an anti-inflammatory agent, an anti-tumor agent, a small molecule drug, or a mixture thereof.

[0058] Further, the nucleic acid comprises at least one of a messenger RNA (mRNA), a small interfering RNA (siRNA), a microRNA, a single guide RNA (sgRNA), an antisense oligonucleotide (ASO), a DNA, and a plasmid.

[0059] Further, the mRNA comprises a tumor antigen mRNA.

[0060] In some specific embodiments, the ganglioside is a compound of Formula (I) or Formula (II), or a pharmaceutically acceptable salt thereof:

[0061]

[0062] wherein,

[0063] R 1selected from the group consisting of monosialyl fucotetrahexose, monosialyl tetrahexose, monosialyl trihexose, monosialyl dihexose, disialyl dihexose, disialyl trihexose, disialyl tetrahexose, or trisialyl dihexose;

[0064] R 2 selected from the group consisting of substituted or unsubstituted straight chain or branched alkanes comprising 7 to 24 carbon atoms.

[0065] Further, at least one carbon atom on said R 2 is optionally replaced by at least one selected from the group consisting of -C(=O)-, -NH-, -O-, -S-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -C(=O)NH-, or -NHC(=O).

[0066] Further, at least one carbon atom on said R 1 is selected from at least one of the following structures:

[0067]

[0068] Further, at least one carbon atom on said R 2 is selected from at least one of the following structures:

[0069]

[0070] In some specific embodiments, the ganglioside is selected from the group consisting of compounds represented by the following structures or a pharmaceutically acceptable salt thereof:

[0071]

[0072] Further, the ganglioside is selected from the group consisting of compounds represented by the following structure (I) or a pharmaceutically acceptable salt thereof:

[0073]

[0074] R 1 is monosialyl fucotetrahexose;

[0075] R 2 is a straight chain alkyl group comprising 10 to 20 carbon atoms.

[0076] In some specific embodiments, the ganglioside is selected from the group consisting of compounds represented by the following structures or a pharmaceutically acceptable salt thereof:

[0077] BRIEF DESCRIPTION OF DRAWINGS

[0078] Figure 1Structure of ganglioside lipid nanoparticle GMLNP encapsulating nucleic acid.

[0079] Figure 2 a~c are the physicochemical properties of mRNA-encapsulated lipid nanoparticles GMLNP in Example 1 (a~c are particle size, encapsulation efficiency and zeta potential, respectively).

[0080] Figure 3 a~g are the in vivo and organ imaging of GMLNP in Example 2 for delivering Fluc mRNA in mice (a~g are lipid nanoparticles formed by different gangliosides, respectively).

[0081] Figure 4 The imaging intensity quantification of GMLNP in Example 2 for delivering Fluc mRNA in mice.

[0082] Figure 5 a~d are the changes of particle size, PDI, encapsulation efficiency and zeta potential of GMLNP in Example 3 when stored at 4 degrees over time.

[0083] Figure 6 a~c are the changes of luminescence intensity in mice after repeated injection of three mFluc-LNPs in Example 4 (a~c are injection strategy, fluorescence imaging and data statistics, respectively).

[0084] Figure 7 a~d are the levels of anti-PEG antibodies and ganglioside antibodies in serum in mice after repeated injection of two mRNA-LNPs in Example 4 (a~d are injection strategy, anti-PEG IgG antibody, anti-PEG IgM antibody and anti-ganglioside antibody levels, respectively).

[0085] Figure 8 a~d are the detection results of blood biochemical indicators (ALT, AST, CR and Urea) at different time points after injection of GMLNP in mice in Example 5.

[0086] Figure 9 The results of GMLNP in Example 6 for delivering hEPO mRNA encoding secreted protein in vivo.

[0087] Figure 10 a~i are the effects of GMLNP in Example 7 for delivering tumor antigen mRNA for treating melanoma (a is the experimental flow design of tumor, b is the tumor growth curve statistical chart of different administration groups, c is the tumor growth curve chart of each mouse in each group, d is the survival rate statistical chart of mice, e is the tumor dissection chart of mice and tumor weight statistical chart, f~h are the tumor-related CD80 + CD86 + macrophages, tumor-related CD80+ CD86 + mature dendritic cells and CD8 + the ratio of T cells, i is a statistical graph of the change in mouse body weight. DETAILED DESCRIPTION

[0088] The present application will be specifically described below in conjunction with specific embodiments and examples, and the advantages and various effects of the present application will be more clearly presented thereby. Those skilled in the art should understand that these specific embodiments and examples are used to illustrate the present application, rather than limit the present application.

[0089] If not specifically stated, all embodiments and optional embodiments of the present application can be combined to form new technical solutions. If not specifically stated, all technical features and optional technical features of the present application can be combined to form new technical solutions. If not specifically stated, all steps of the present application can be performed in sequence, or randomly, and preferably in sequence. For example, the method comprises steps (a) and (b), which means that the method can comprise steps (a) and (b) performed in sequence, or steps (b) and (a) performed in sequence. For example, the method can also comprise step (c), which means that step (c) can be added to the method in any order, for example, the method can comprise steps (a), (b) and (c), or steps (a), (c) and (b), or steps (c), (a) and (b), etc.

[0090] If not specifically stated, the terms used in the present application have the commonly understood meanings understood by those skilled in the art.

[0091] The terms used herein are for the purpose of describing specific embodiments only and are not intended to limit the present disclosure. As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and / or "comprising," when used in this specification, specify the presence of stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0092] In the description herein, it should be noted that, unless otherwise specified, "above", "below" include the number itself, and the meaning of "more" in "one or more" is more than two.

[0093] Lipid nanoparticles (LNP) and liposomes are both lipid-based delivery systems, but they have significant differences in structure, composition, and application. Compared with liposomes, the structure and composition of LNP are more complex. The composition of liposomes is relatively simple, mainly phospholipids and cholesterol. Liposomes are mainly used for the delivery of water-soluble drugs and some small molecule drugs. While LNP is mainly used for the delivery of nucleic acid drugs, especially mRNA vaccines and mRNA drugs.

[0094] Example 1, Preparation and characterization of ganglioside lipid nanoparticle GMLNP

[0095] Cationic lipid SM102 was mixed with DSPC, cholesterol, and ganglioside in a molar ratio of 50:10:38.5:1.5 and dissolved in anhydrous ethanol, with a total lipid concentration of 10 mM. Luciferase mRNA (Fluc mRNA) was dissolved in a sodium acetate aqueous solution (50 mM, pH = 5.0) with a mRNA concentration of 0.15 mg / mL. Using a microfluidic instrument, the flow rate ratio of ethanol solution to sodium acetate aqueous solution was controlled at 1:3, and the solution of lipid nanoparticles was prepared in a microfluidic chip. The ethanol was removed by repeated centrifugation at 4°C with PBS dilution three times, and the resulting concentrate was diluted with PBS for standby use. The size and polydispersity index (PDI) of the lipid nanoparticles were determined by dynamic light scattering using a Malvern Zetasizer Nano ZS in 173° backscattering detection mode. The encapsulation efficiency of the lipid nanoparticles was determined using the Quant-it Ribogreen RNA quantification assay kit (ThermoFisher Scientific). The final size, PDI, zeta potential (i.e., zeta potential), and encapsulation efficiency of the resulting lipid nanoparticles are shown in Table 1 and Figure 1

[0096] Table 1

[0097]

[0098]

[0099] From Table 1 and Figure 2 ​As can be seen from a~c, the GMLNPs prepared from the ganglioside lipids prepared in Example 1 can encapsulate mRNA molecules, the obtained mRNA-LNP has a suitable particle size of 110~190 nm, good uniformity with a PDI less than 0.15, a zeta potential of -7~-13 mV, and an encapsulation efficiency greater than 90%, among which GD3CerC18, GT3CerC18, FucGM1CerC14 and FucGM1CerC18 have the highest encapsulation efficiency (all greater than 98%). Thus, the ganglioside lipids can completely replace PEG lipids to form stable LNPs.

[0100] Example 2, mouse imaging experiment of ganglioside lipid nanolipid particle GMLNP

[0101] Six to eight-week-old female BALB / c mice with a body weight of about 20 g were selected, and the feeding environment was a SPF level feeding room. Animal experiments were strictly in accordance with the guidelines of the national health organization and the requirements of animal ethics. Specifically, mRNA encoding luciferase was prepared into different ganglioside LNPs as described above, and PEG-LNPs were used as a control group. Three mice were randomly selected from each group, and the mice were injected with mRNA at a dose of 0.5 mg / kg via the tail vein. Six hours after injection of LNP, 200 μL of 10 mg / mL D-luciferin potassium salt was injected intraperitoneally into each mouse, and after 10 minutes, the mice were placed under a live imaging system (PerkinElmer IVIS Spectrum) to collect whole body and specific organ samples and monitor bioluminescence signals. The experimental results show that Figure 3 a~g and Figure 4 ), compared with PEG-LNPs, the bioluminescence imaging signal intensity of all kinds of ganglioside LNPs in the spleen was significantly improved, and the signal intensity in the liver was lower, indicating that the ganglioside LNPs have excellent performance in targeted delivery to the spleen. Among these glycolipids, FucGM1CerC14 and FucGM1CerC18 have the highest delivery efficiency.

[0102] Example 3, stability study of GMLNP

[0103] The stability of ganglioside LNPs was determined by monitoring the nanoparticle size, PDI, encapsulation efficiency and zeta potential of ganglioside LNPs over a period of time. Taking FucGM1CerC18 as an example, the particle size, PDI, encapsulation efficiency and zeta potential of the ganglioside LNPs were monitored at 4℃ for 2 weeks. Figure 5 As can be seen from a~d, the ganglioside LNPs stored at 4℃ have very small changes in their parameters within two weeks, proving that they have good stability.

[0104] Example 4, repeated administration study of GMLNP

[0105] BALB / c mice (6-8 weeks old, n=3) were injected with Luciferase mRNA (mFluc) containing PEG-LNP and FucGM1CerC18-LNP via tail vein at a dose of 0.5 mg / kg mRNA, with an interval of 4 days for 3 times. The mice were euthanized 6 h after the 1st and 3rd injection, and dissected and imaged. The imaging results showed that Figure 6 a-c), after 3 repeated administrations, the bioluminescence intensity of the PEG-LNP group decreased significantly by 91.7%, while the bioluminescence of the FucGM1CerC18-LNP group remained at the same level. This indicates that the ganglioside LNP can maintain the efficacy of mRNA vaccine or drug better than PEG-LNP when repeatedly administered.

[0106] BALB / c mice (6-8 weeks old, n=3) were injected with Luciferase mRNA containing FucGM1CerC18-LNP and PEG-LNP via tail vein, with the cationic lipids SM102 and MC3, at a dose of 0.5 mg / kg mRNA, with an interval of 1 week for 2 times. Blood was collected 1 week after the 2nd injection for ELISA experiments to determine the levels of anti-PEG IgG and IgM antibodies and anti-FucGM1CerC18 IgM antibodies in the serum of mice after two administrations. The results are shown in Figure 7 a-d, no significant anti-PEG antibody signal was detected in the serum of FucGM1CerC18-LNP treated mice, while the PEG-LNP treated mice showed a significant increase in anti-PEG IgG and IgM levels in the serum one week after the second administration. To evaluate whether the ganglioside LNPs of the present application would trigger an immune response to gangliosides, we detected the levels of anti-FucGM1CerC18 antibodies in the serum of mice. The results showed that no anti-FucGM1CerC18 antibodies were detected in the serum of mice injected with FucGM1CerC18-LNPs, which showed no significant difference compared with the untreated mouse (UT) group, proving that the ganglioside LNPs of the present application significantly reduce the generation of anti-PEG antibodies compared with PEG-LNPs, and do not trigger an immune response to gangliosides, thereby having higher safety and the advantage of being suitable for repeated administration.

[0107] Due to the transient nature of mRNA expression, repeated dosing is necessary to achieve optimal therapeutic effect. The results of this experiment also demonstrate that repeated dosing of PEG-modified LNPs induces high levels of anti-PEG antibody production and attenuates their therapeutic effect. The ganglioside LNPs of the present application, by using ganglioside to stabilize the lipid nanoparticles, do not produce anti-PEG antibodies and anti-ganglioside antibodies, avoiding the adverse reactions of PEG-LNPs, and are more suitable for repeated dosing.

[0108] Example 5, Safety study of GM LNP

[0109] The present application detected the biochemical indicators of mouse blood, including alanine aminotransferase ALT, aspartate aminotransferase AST, creatinine CR and urea Urea. The results show that Figure 8 a~d), compared with the PBS injection group, the biochemical indicators of FucGM1CerC18-LNP treated mice had no significant change, proving that the ganglioside LNPs of the present application have good safety.

[0110] Example 6, In vivo delivery of mRNA encoding secreted protein hEPO by GM LNP

[0111] BALB / c mice (6-8 weeks old, n=3) were injected with FucGM1CerC18-LNP and PEG-LNP containing hEPO mRNA in the tail vein, and the cationic lipid used was SM102, the dose was 0.5 mg / kg mRNA. Blood was collected at different time points (3h, 6h, 12h, 24h, 48h, 72h) after injection. According to the manufacturer's instructions, ELISA analysis was performed using a commercially available kit (DEP00, R&D Systems), and the test results Figure 9 ) show that FucGM1CerC18-LNP can produce more hEPO secreted protein than PEG-LNP in vivo, further proving that FucGM1CerC18-LNP has higher delivery efficiency.

[0112] Example 7, Application of GM LNP in tumor mRNA vaccine

[0113] mRNA tumor vaccine is a new type of tumor immunotherapy drug, and its basic principle is to deliver mRNA encoding tumor-specific antigen targets into the body through a specific way. After these mRNAs enter human cells, they use the protein synthesis mechanism of human cells to produce tumor antigens, thereby triggering an immune response. Melanoma is a highly malignant tumor derived from melanocytes, and the incidence and mortality of malignant melanoma have been increasing year by year in recent years. In addition to early surgical resection, there is no specific treatment for malignant melanoma, and the prognosis is poor. Therefore, it is extremely important to develop an mRNA tumor vaccine for melanoma. Melanoma-associated antigen 3 (MAGEA3) is highly expressed in melanoma patients, and the present application verifies that GMLNP delivery of MAGEA3-encoding mRNA can efficiently inhibit tumor growth.

[0114] B16F10 melanoma cells (2 x 10 5 ) were subcutaneously injected into the lateral thigh of mice. On the 6th, 10th and 14th day after injection, each group of mice was intravenously injected with PBS, LNP group using PEG-wrapped MAGEA3 mRNA (PEG-MA), LNP group using FucGM1CerC18 ganglioside-wrapped MAGEA3 mRNA (FucGM-MA), and LNP control group using FucGM1CerC18 ganglioside-wrapped Luciferase mRNA (FucGM-Luc), with an mRNA dose of 0.5 mg / kg. When the tumor volume reached 1500 mm 3 , the mice were euthanized. As shown in Figure 10 a-i, compared with the PEG-MA group, the FucGM-MA group of mice had the best tumor inhibition effect, with the smallest tumor volume and weight, and the FucGM-MA group of mice also showed the longest survival time. After analyzing the immune cells in the tumor tissue, it was found that the proportion of tumor-associated macrophages (TAMs) (CD80 + CD86 + macrophages), mature dendritic cells (DCs) (CD80 + CD86 + ), and CD8 + T cells increased in the FucGM-MA group, all of which indicated that the anti-tumor immune response in the tumor microenvironment (TME) was more effectively activated. The body weight of the FucGM-MA group of mice also did not change significantly, which proved that it was also very safe.

[0115] The results of Example 7 proved that the GMLNP composed of FucGM1CerC18 ganglioside can more effectively stimulate the anti-tumor immune response in vivo compared with PEG-LNP, and has a better effect on the delivery of mRNA tumor vaccine in vivo.

[0116] Note that the present application is not limited to the above-described embodiments. The above-described embodiments are merely examples, and embodiments having substantially the same configuration, function, and effect as the technical idea of the present application are included in the technical scope of the present application. Furthermore, other modes constructed by applying various modifications to the embodiments, or by combining part of the configurations of the embodiments, which can be conceived by those skilled in the art, without departing from the spirit of the present application, are also included in the scope of the present application.

Claims

1. A lipid nanoparticle comprising a ganglioside, a cationic lipid, a helper phospholipid and a sterol lipid.

2. The lipid nanoparticle of claim 1, wherein, The ganglioside is a compound of Formula (I) or Formula (II) or a pharmaceutically acceptable salt thereof: wherein, R 1 selected from monosialylated fucosetetrahexose, monosialylated tetrahexose, monosialylated trihexose, monosialylated dihexose, disialylated dihexose, disialylated trihexose, disialylated tetrahexose, or trisialylated dihexose; R 2 selected from the group consisting of substituted or unsubstituted straight or branched chain alkanes containing from 7 to 24 carbon atoms.

3. The lipid nanoparticle of claim 1 or 2, wherein, The ganglioside is selected from a compound of the following structure (I) or a pharmaceutically acceptable salt thereof: R 1 is a monosialyl fucosetetrahexose; R 2 R is a straight-chain alkyl group containing 10 to 20 carbon atoms.

4. The lipid nanoparticle of claim 3, wherein, The ganglioside is selected from a compound of the following structure (I) or a pharmaceutically acceptable salt thereof: R 1 is a monosialyl fucosetetrahexose; R 2 R is a straight-chain alkyl group containing 13 or 17 carbon atoms.

5. The lipid nanoparticle of any one of claims 1-4, wherein, The cationic lipid comprises one or more of SM102 or MC3; the helper phospholipid comprises one or more of phosphatidylcholine DSPC, phosphatidylethanolamine DOPE; and / or the sterol lipid comprises at least one of cholesterol, beta-sitosterol, ergosterol, campesterol, stigmasterol, a corticosteroid.

6. The lipid nanoparticle of any one of claims 1-5, wherein, The molar ratio between the cationic lipid, the helper phospholipid, the sterol lipid, the ganglioside is (30-60):(5-25):(25-50):(0.05-10).

7. A composition comprising the lipid nanoparticle of any one of claims 1-6, wherein, The lipid nanoparticle is loaded with an active agent.

8. The composition of claim 7, wherein, The active agent is mRNA.

9. Use of the lipid nanoparticle of any one of claims 1-6 or the composition of claim 7 or 8 for the manufacture of a medicament.

10. A method of preparing a lipid nanoparticle, comprising: providing a ganglioside, a cationic lipid, a helper phospholipid and a sterol lipid; mixing and dissolving the above substances to obtain a mixture; and processing the mixture to obtain a lipid nanoparticle.

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