Chloroquine lipid, preparation method and application thereof, and lipid nanoparticles containing chloroquine lipid

Abstract

The invention relates to a biological medicine technology, and discloses chloroquine lipid, a preparation method and application thereof, and lipid nanoparticles containing the chloroquine lipid. The chloroquine lipid is synthesized from chloroquine substances and alkylene oxide through a ring-opening reaction, and the chloroquine substances are selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives and hydroxychloroquine derivatives. The lipid nanoparticles contain the chloroquine lipid, cationic lipid, phospholipid, PEG (polyethylene glycol) lipid and cholesterol. The chloroquine lipid can be used as a lipidated autophagy inhibitor, can endow the LNPs carrier with an autophagy inhibition function, and realizes the autophagy inhibition effect of'time-space synchronization ', so that the obstacle of'autophagy lysosome degradation' is broken through, and the gene delivery efficiency is enhanced.

Description

Technical Field

[0001] The present invention relates to biomedical technology, and specifically, to a chloroquine lipid, a preparation method and application thereof, and a lipid nanoparticle containing the chloroquine lipid. Background Art

[0002] Lipid nanoparticles (LNPs) are a commonly used and advanced non-viral gene drug delivery system, usually composed of four lipids: ionizable lipids, helper lipids, cholesterol, and polyethylene glycolylated lipids. Although LNPs play a key role in the progress of emerging gene therapy drugs, there are still key problems such as low delivery efficiency and low endosome / lysosome escape performance, which result in far from meeting the urgent needs of clinical research and gene therapy.

[0003] Cell autophagy is an evolutionary mechanism of cell self-degradation in eukaryotic systems. In response to different stimuli / stresses, including starvation, high temperature, hypoxia, and xenobiotics, damaged, denatured, or aged proteins, organelles, and foreign substances in cells are transported to lysosomes for digestion and degradation. A large number of studies have confirmed that autophagy is an obstacle to gene transfection. The process of LNPs carrying gene drugs escaping and releasing from endosomes / lysosomes promotes the formation of autophagy, and autophagy then brings gene drugs into the autophagy-lysosome degradation pathway, which greatly reduces the transfection efficiency of gene drugs.

[0004] How to effectively break through the "autophagy-lysosome degradation" barrier in the process of LNPs delivering drugs has become a key technical problem in the development of LNPs. The existing technologies mainly focus on the development of ionizable lipids in LNPs, and use the improvement of the performance of ionizable lipids to help LNPs escape endosomes to improve delivery efficiency. However, these technologies cannot avoid the negative impact of autophagy-lysosome degradation on delivery. Summary of the Invention

[0005] The object of the present invention is to overcome the problem that autophagy is formed during the process of LNPs delivering gene drugs in the prior art, which greatly reduces the transfection efficiency of gene drugs. The present invention provides a chloroquine lipid, a preparation method and application thereof, and a lipid nanoparticle containing the chloroquine lipid. The chloroquine lipid can be used as a lipidated autophagy inhibitor, which can endow the LNPs carrier with its own autophagy inhibition function, realize the "spatiotemporal synchronous" autophagy inhibition effect, thereby breaking through the "autophagy-lysosome degradation" barrier and enhancing the gene delivery efficiency.

[0006] To achieve the above object, in the first aspect of the present invention, a chloroquine lipid is provided. The chloroquine lipid is synthesized by a ring-opening reaction of a chloroquine substance and an alkylene oxide, and the chloroquine substance is selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives, and hydroxychloroquine derivatives.

[0007] Preferably, the structural formula of the chloroquine lipid is as shown in formula (I),

[0008]

[0009] wherein, m is an integer from 1 to 20, and R1 and R2 are each independently selected from hydrogen or hydroxyl.

[0010] Preferably, R1 and R2 are hydrogen, and m is an integer from 5 to 13.

[0011] In a second aspect of the present invention, a method for preparing a chloroquine lipid is provided, the method comprising: mixing a chloroquine substance with an alkylene oxide for a ring-opening reaction in the presence of a reaction solvent; wherein, the chloroquine substance is selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives and hydroxychloroquine derivatives.

[0012] Preferably, the chloroquine substance is chloroquine and / or hydroxychloroquine, preferably chloroquine.

[0013] Preferably, the structural formula of the alkylene oxide is as shown in formula (II),

[0014] n is an integer from 4 to 23.

[0015] Preferably, the molar ratio of the chloroquine substance to the alkylene oxide is 1:0.9 - 1.1.

[0016] Preferably, the conditions of the ring-opening reaction include: temperature is 80 - 90 °C, and time is 40 - 60 h.

[0017] Preferably, the reaction solvent is a lower organic alcohol, more preferably isopropanol.

[0018] Preferably, the method further comprises: adjusting the pH of the chloroquine substance to 6.5 - 7.5 before mixing the chloroquine substance with the alkylene oxide.

[0019] In a third aspect of the present invention, the application of the chloroquine lipid as described above and / or the chloroquine lipid prepared by the method as described above in the preparation of lipid nanoparticles is provided.

[0020] In a fourth aspect of the present invention, a lipid nanoparticle is provided, the lipid nanoparticle contains chloroquine lipid, cationic lipid, phospholipid, PEG lipid and cholesterol; wherein, the chloroquine lipid is the chloroquine lipid as described above and / or the chloroquine lipid prepared according to the method as described above.

[0021] Preferably, the molar ratio of the chloroquine lipid, cationic lipid, phospholipid, PEG lipid and cholesterol is (2-15):(15-30):(3-10):(20-30):1, more preferably (3-15):(20-30):(3-10):(22-28):1.

[0022] Preferably, the molar ratio of the chloroquine lipid to the cationic lipid is 0.1-0.4:1.

[0023] Preferably, the cationic lipid is selected from at least one of SM-102, MC3 and ALC-0315, the phospholipid is DSPC, and the PEG lipid is DMG-PEG2000 and / or ALC-0159.

[0024] The fifth aspect of the present invention provides the use of the lipid nanoparticles as described above in delivering gene drugs.

[0025] Through the above technical solutions, the beneficial effects of the present invention are as follows: The present invention provides a new chloroquine lipid, which can be used as a lipidated autophagy inhibitor and introduced into the LNP system to construct a new type of highly efficient autophagy lysosome degradation-inhibiting LNP delivery platform for gene delivery. By loading autophagy inhibitory groups, the AUDI-LNPs vector itself has autophagy inhibitory function, realizing "spatiotemporal synchronous" autophagy inhibition, thereby breaking through the "autophagy lysosome degradation" barrier and enhancing gene delivery efficiency. Description of the Drawings

[0026] Figure 1 It is the fluorescence intensity diagram of AUDI-LNP delivering Luc mRNA formed by different chloroquine lipids (CQn) in Example 2;

[0027] Figure 2 It is the fluorescence intensity diagram of AUDI-LNP delivering Luc mRNA formed by different proportions of chloroquine lipid (CQ12) in Example 3;

[0028] Figure 3 It is the fluorescence intensity diagram of AUDI-LNP delivering LucmRNA formed by applying chloroquine lipid (CQ12) to different LNP systems in Example 4;

[0029] Figure 4 It is the in vivo imaging diagram of each group of mice in Example 5;

[0030] Figure 5 It is the quantitative result diagram of the in vivo imaging of each group of mice in Example 5. Detailed Embodiments

[0031] The endpoints and any values in the ranges disclosed herein are not limited to the exact ranges or values, and these ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0032] The first aspect of the present invention provides a chloroquine lipid, which is synthesized by the ring-opening reaction of a chloroquine substance and an alkylene oxide, and the chloroquine substance is selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives and hydroxychloroquine derivatives.

[0033] During the R & D process, the inventors of the present invention unexpectedly found that by subjecting a chloroquine substance to a ring-opening reaction with an alkylene oxide to lipidize chloroquine to form a chloroquine lipid, it can be used as a lipidized autophagy inhibitor and introduced into the LNP system to construct a novel and highly efficient autolysosomal degradation-inhibitory LNPs delivery platform (Autolysosomal degradation-inhibitory LNPs, AUDI-LNPs). By loading autophagy inhibitory groups, the AUDI-LNPs carrier itself is endowed with autophagy inhibitory function, realizing "spatiotemporal synchronous" autophagy inhibition, thus breaking through the "autolysosomal degradation" barrier, avoiding autolysosomal degradation, and enhancing gene delivery efficiency.

[0034] According to the present invention, the chloroquine substance can be chloroquine, hydroxychloroquine, any chloroquine derivative or any hydroxychloroquine derivative; the alkylene oxide can be any alkane substance containing an epoxy group. Exemplarily, the structural formula of the alkylene oxide is shown as formula (II),

[0035] n can be any integer greater than 3.

[0036] According to the present invention, preferably, the chloroquine substance is chloroquine and / or hydroxychloroquine, and the structural formula of the alkylene oxide is shown as formula (II). At this time, the structural formula of the chloroquine lipid is shown as formula (I),

[0037]

[0038] Wherein, m is an integer from 1 to 20, and R1 and R2 are each independently selected from hydrogen or hydroxyl; further preferably, R1 and R2 are hydrogen, and m is an integer from 5 to 13. In this preferred embodiment, it is beneficial to further enhance the autophagy inhibitory function of the LNP carrier and enhance gene delivery efficiency.

[0039] The second aspect of the present invention provides a method for preparing chloroquine lipids, which includes: mixing a chloroquine substance with an alkylene oxide in the presence of a reaction solvent for a ring-opening reaction; wherein, the chloroquine substance is selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives and hydroxychloroquine derivatives.

[0040] The method for preparing chloroquine lipids provided by the present invention is simple. After ring-opening with an alkylene oxide and bonding to the secondary amino group in the chloroquine substance, the chloroquine substance is lipidated and further introduced into the LNP system as a lipidated autophagy inhibitor.

[0041] According to the present invention, preferably, the chloroquine substance is chloroquine and / or hydroxychloroquine, preferably chloroquine.

[0042] According to the present invention, preferably, the structural formula of the alkylene oxide is shown in formula (II),

[0043] n is an integer from 4 to 23. In this preferred embodiment, it is beneficial to further enhance the autophagy inhibitory function of the LNP carrier and improve the gene delivery efficiency.

[0044] According to the present invention, preferably, the molar ratio of the chloroquine substance to the alkylene oxide is 1:0.9 - 1.1, specifically it can be 1:0.9, 1:0.95, 1:1, 1:1.05, 1:1.1, or any value between the above two values.

[0045] According to the present invention, preferably, the conditions of the ring-opening reaction include: the temperature is 80 - 90 °C, specifically it can be 80 °C, 82 °C, 84 °C, 86 °C, 88 °C, 90 °C, or any value between the above two values; the time is 40 - 60 h, specifically it can be 40 h, 45 h, 50 h, 55 h, 60 h, or any value between the above two values. In this preferred embodiment, it is beneficial to improve the synthesis efficiency of chloroquine lipids.

[0046] According to the present invention, preferably, the reaction solvent is a lower organic alcohol, such as ethanol, n-propanol, isopropanol, n-butanol, isobutanol, etc., more preferably isopropanol. In this preferred embodiment, it is beneficial to improve the synthesis efficiency of chloroquine lipids.

[0047] According to the present invention, preferably, the method further includes: before mixing the chloroquine substance with the alkylene oxide, adjusting the pH of the chloroquine substance to 6.5 - 7.5. Specifically, the chloroquine substance can be first dissolved in the reaction solvent, and then the pH is adjusted to 6.5 - 7.5 using a pH regulator to neutralize hydrochloric acid.

[0048] In the present invention, the preparation method of chloroquine lipid further includes: purifying the product of the ring-opening reaction to obtain chloroquine lipid; exemplarily, chromatographic purification is carried out using a mixed solution of MeOH and ammonia water with a concentration of 0.05 - 0.2 wt%, and further preferably, the volume ratio of MeOH to ammonia water with a concentration of 0.05 - 0.2 wt% is 1 - 1.1:1.

[0049] Based on the fact that the chloroquine lipid provided by the present invention has the effect of inhibiting autophagolysosome degradation, the third aspect of the present invention provides the application of the aforementioned chloroquine lipid and / or the chloroquine lipid prepared by the aforementioned method in the preparation of lipid nanoparticles.

[0050] The fourth aspect of the present invention provides a lipid nanoparticle, which contains chloroquine lipid, cationic lipid, phospholipid, PEG lipid and cholesterol; wherein, the chloroquine lipid is the aforementioned chloroquine lipid and / or the chloroquine lipid prepared according to the aforementioned method. This lipid nanoparticle has an autophagy inhibition function, realizes "spatiotemporal synchronous" autophagy inhibition, thus breaking through the "autophagolysosome degradation" barrier and enhancing gene delivery efficiency.

[0051] According to the present invention, preferably, the molar ratio of the chloroquine lipid, cationic lipid, phospholipid, PEG lipid and cholesterol is (2 - 15):(15 - 30):(3 - 10):(20 - 30):1, and more preferably (3 - 15):(20 - 30):(3 - 10):(22 - 28):1.

[0052] According to the present invention, preferably, the molar ratio of the chloroquine lipid to the cationic lipid is 0.1 - 0.4:1, specifically it can be 0.1:1, 0.2:1, 0.3:1, 0.4:1, or any value between the above two values.

[0053] According to the present invention, preferably, the cationic lipid is selected from at least one of SM - 102, MC3 and ALC - 0315, the phospholipid is DSPC, and the PEG lipid is DMG - PEG2000 and / or ALC - 0159.

[0054] The present invention can be based on any existing lipid nanoparticle system, use chloroquine lipid to replace part of its cationic lipid, and adopt the existing or self - designed LNPs preparation method to form a new autophagolysosome degradation - inhibiting LNPs delivery platform, realize "spatiotemporal synchronous" autophagy inhibition, thus breaking through the "autophagolysosome degradation" barrier and enhancing gene delivery efficiency.

[0055] The fifth aspect of the present invention provides the application of the aforementioned lipid nanoparticle in delivering gene drugs.

[0056] In the present invention, when lipid nanoparticles are used to deliver gene drugs, the lipid nanoparticles and gene drugs are preferably delivered in a system with a mass ratio of 15-25:1, which is beneficial to further improve the delivery efficiency of the gene drugs.

[0057] The present invention will be described in detail below through examples.

[0058] In the following examples, the CAS number of SM-102 is 2089251-47-6, purchased from Avanti Lipids (Shanghai) Pharmaceutical Technology Co., Ltd., product model SM220810; the CAS number of DSPC is 816-94-4, purchased from Avanti Lipids Polar Inc., product model 850365P; the CAS number of cholesterol is 57-88-5, purchased from Sigma-Aldrich, product model C8667; the CAS number of DMG-PEG2000 is 160743-62-4, purchased from Avanti Lipids Polar Inc., product model: 880151P; MC3, CAS number: 1224606-06-7, purchased from Avituo (Shanghai) Pharmaceutical Technology Co., Ltd., product model: O02006; ALC-0315, CAS number: 2036272-55-4, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., product model: A419992; ALC-0159, CAS number: 1849616-42-7, purchased from Shanghai Haohong Biopharmaceutical Technology Co., Ltd., product model: 1218006;

[0059] Unless otherwise specified, the remaining raw materials or reagents are conventional commercially available products.

[0060] Example 1 Synthesis of Chloroquine Lipids

[0061] Based on the structure of chloroquine (CQ), chloroquine lipid (CQn) was synthesized through the ring-opening reaction of 1,2-epoxyalkylene.

[0062] The specific process is as follows: 0.3126 mmol of CQ was dissolved in 5 mL of isopropanol and placed in a glass bottle equipped with a magnetic stirring bar; before adding 0.32 mmol of 1,2-epoxyalkane, 0.18 mmol of potassium carbonate was added to neutralize the hydrochloric acid; then the vial was sealed and stirred at 85°C for 48 hours, and the crude product was purified by chromatography using a MeOH / 0.1% NH4OH solvent mixture with a volume ratio of 52:48 to obtain chloroquine lipid.

[0063] The synthetic route of chloroquine lipid is shown in the following reaction formula,

[0064]

[0065] Among them, n takes values of 6, 8, 10, 12, 14, 16, and 18 respectively;

[0066] The corresponding CQn was characterized by mass spectrometry, and the results are shown in Table 1.

[0067] Table 1 Mass spectrometry characterization of CQn

[0068]

[0069]

[0070] Example 2

[0071] (1) Using SM-102 / DSPC / cholesterol / DMG-PEG2000 (molar ratio of

[0072] 50.0:10.0:38.5:1.5) as the basic LNP, various chloroquine lipids (CQn) prepared in Example 1 were used to replace the ionizable lipid SM-102 in the basic LNP at a molar ratio of 30% respectively, so that the molar ratio of the five components of CQn / SM-102 / DSPC / cholesterol / DMG-PEG2000 was 15.0:35.0:10.0:38.5:1.5, forming a new mixed lipid (AUDI-LNP);

[0073] (2) Take 100 μg of the new mixed lipid and dissolve it in 25 μL of ethanol as the organic phase; take 5 μg of Luc mRNA (as the reporter gene, purchased from Suzhou Novoprotein Science & Technology Co., Ltd., product model MR009) and dissolve it in 75 μL of citrate buffer solution (pH 4.0) as the aqueous phase; quickly inject 25 μL of the organic phase into 75 μL of the aqueous phase, so that the mass ratio of the mixed lipid to the nucleic acid drug is 20:1, and the volume ratio of the organic phase to the aqueous phase is 1:3; and stir magnetically at a speed of 1500 revolutions for 3 minutes to obtain a mixed solution;

[0074] (3) Subsequently, dilute the above mixed solution with 20 times the volume of PBS buffer solution, place it in a 30 kD centrifugal ultrafiltration tube, and centrifuge at 3500 g for 15 minutes to obtain 100 μL of LNPs solution encapsulating Luc mRNA; after treating HEK293T cells with the LNPs solution encapsulating Luc mRNA (calculated according to 200 ng of Luc mRNA) for 12 h, discard the cell supernatant, and sequentially add lysis buffer (lyse for 5 min) and luciferase substrate reagent (firefly luciferase reporter gene detection kit, purchased from Beyotime Biotechnology Co., Ltd., product model RG005), and immediately detect its fluorescence intensity at 560 nm with an enzyme-labeled instrument to evaluate its delivery efficiency. The results are shown in Figure 1, it was screened out that chloroquine lipids CQ8, CQ10, CQ12, CQ14, and CQ16 were introduced as the fifth component into LNP to form a new LNP delivery platform, which had a high delivery efficiency for nucleic acid drugs. Among them, chloroquine lipid CQ12 had the best efficiency when introduced as the fifth component into LNP to form a new LNP delivery platform.

[0075] Example 3

[0076] Using SM-102 / DSPC / cholesterol / DMG-PEG2000 (molar ratio of 50.0:10.0:38.5:1.5) as the basic LNP, and using chloroquine lipid CQ12 prepared in Example 1, it replaced the ionizable lipid SM-102 in LNP at molar ratios of 0%, 1%, 2%, 5%, 10%, 15%, 20%, 30%, 40%, 50%, 75%, and 100% respectively, so that the molar ratios of the five components of CQn / SM-102 / DSPC / cholesterol / DMG-PEG2000 were 0:50:10.0:38.5:1.5, 0.5:49.5:10.0:38.5:1.5, 1:49:10.0:38.5:1.5, 2.5:47.5:10.0:38.5:1.5, 5:45:10.0:38.5:1.5, 7.5:42.5:10.0:38.5:1.5, 10:40:10.0:38.5:1.5, 15:35:10.0:38.5:1.5, 20:30:10.0:38.5:1.5, 25:25:10.0:38.5:1.5, 35:15:10.0:38.5:1.5, 50:0:10.0:38.5:1.5, respectively, to form a new mixed lipid (AUDI-LNP);

[0077] Using the steps (2) and (3) described in Example 2, an LNPs solution encapsulating Luc mRNA was prepared, and the fluorescence intensity was detected to evaluate its delivery efficiency. The results are shown in Figure 2 .

[0078] It was screened out that chloroquine lipid CQ12 replaced the ionizable lipid SM-102 in LNP at molar ratios of 10%, 15%, 20%, 30%, and 40% respectively to form a new LNP delivery platform, which had a high delivery efficiency for nucleic acid drugs. Among them, chloroquine lipid CQ12 had the best efficiency when introduced into LNP by replacing the ionizable lipid SM-102 in LNP at a molar ratio of 30%.

[0079] Example 4 Compatibility of AUDI technology with different LNPs formulations

[0080] Currently, there are 3 kinds of LNPs approved by the FDA: LNPs MC3, LNPs SM-102 and LNPs ALC-0315 , wherein,

[0081] The formulation of LNPs MC3 is MC3 / DSPC / cholesterol / DMG-PEG2000 (the molar ratio of each component is 50.0:10.0:38.5:1.5),

[0082] The formulation of LNPs SM-102 is SM-102 / DSPC / cholesterol / DMG-PEG2000 (the molar ratio of each component is 50.0:10.0:38.5:1.5),

[0083] The formulation of LNPs ALC-0315 is ALC-0315 / DSPC / cholesterol / ALC-0159 (the molar ratio of each component is 46.3:9.4:42.7:1.6);

[0084] These 3 kinds of LNPs are respectively introduced with AUDI technology, that is, chloroquine lipid CQ12 is used to replace the ionizable lipid MC3, SM-102 or ALC-0315 in LNPs at a molar ratio of 30%, and step (1) described in Example 2 is adopted to form a new mixed lipid AUDI-LNPs MC3 , AUDI-LNPs SM-102 and AUDI-LNPs ALC-0315 ;

[0085] The LNPs solution encapsulating Luc mRNA is prepared by adopting steps (2) and (3) described in Example 2, and the fluorescence intensity is detected to evaluate its delivery efficiency. The results are shown in Figure 3 .

[0086] It can be seen from Figure 3 that the transfection efficiencies of AUDI-LNPs MC3 , AUDI-LNPs SM-102 and AUDI-LNPs ALC-0315 are significantly higher than those of LNPs MC3 , LNPs SM-102 and LNPs ALC-0315 respectively.

[0087] Example 5 Evaluation of the subcutaneous injection performance of AUDI technology in mice

[0088] Adopt step (1) described in Example 2, and use chloroquine lipid CQ12 to replace the ionizable lipid SM-102 in LNPs SM-102 at a molar ratio of 30% to form a new mixed lipid AUDI-LNPs SM-102;Prepare AUDI-LNPs encapsulating Luc mRNA using steps (2) and (3) described in Example 2 SM-102 solution and LNPs encapsulating Luc mRNA SM-102 solution;

[0089] Select 9 mice (purchased from SPF (Beijing) Biotechnology Co., Ltd., with a specification of 18 - 20 g), and divide them into 3 groups: experimental group, control group, and blank group, with 3 mice in each group; Administer the drug subcutaneously on the small back of the mice. For the experimental group (AUDI-LNPs encapsulating LucmRNA SM-102 ), the dosage for each mouse is 5 μg (calculated based on Luc mRNA). For the control group (LNPs encapsulating Luc mRNA SM-102 ), the dosage for each mouse is 5 μg (calculated based on Luc mRNA). The mice in the blank group are not administered as a control (Control). Observe the fluorescence intensity in a small animal in vivo imaging system, and the results are as shown in Figure 4 and Figure 5 ; At 2 h after administration, there was no significant difference in the fluorescence intensity in the experimental group and the control group. However, at 12 h after administration, the fluorescence intensity in the experimental group was significantly higher than that in the control group.

[0090] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited thereto. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention, including any other suitable combination of each technical feature. These simple modifications and combinations should also be regarded as the content disclosed by the present invention and fall within the protection scope of the present invention.

Claims

1. A chloroquine lipid, characterized in that, The chloroquine lipid is synthesized by the ring-opening reaction of a chloroquine substance and an alkylene oxide, and the chloroquine substance is selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives, and hydroxychloroquine derivatives.

2. The chloroquine lipid according to claim 1, characterized in that, The structural formula of the chloroquine lipid is shown in formula (I), wherein, m is an integer from 1 to 20, and R1 and R2 are each independently selected from hydrogen or a hydroxyl group; Preferably, R1 and R2 are hydrogen, and m is an integer from 5 to 13.

3. A preparation method of chloroquine lipid, characterized in that, The method includes: mixing a chloroquine substance and an alkylene oxide in the presence of a reaction solvent for a ring-opening reaction; wherein, the chloroquine substance is selected from at least one of chloroquine, hydroxychloroquine, chloroquine derivatives, and hydroxychloroquine derivatives.

4. The preparation method according to claim 3, characterized in that, The chloroquine substance is chloroquine and / or hydroxychloroquine, preferably chloroquine; Preferably, the structural formula of the alkylene oxide is shown in formula (II), n is an integer from 4 to 23; Preferably, the molar ratio of the chloroquine substance to the alkylene oxide is 1:0.9 - 1.1; Preferably, the conditions of the ring-opening reaction include: temperature is 80 - 90 °C, and time is 40 - 60 h; Preferably, the reaction solvent is a lower organic alcohol, more preferably isopropanol.

5. The preparation method according to claim 3 or 4, characterized in that, The method further includes: adjusting the pH of the chloroquine substance to 6.5 - 7.5 before mixing the chloroquine substance and the alkylene oxide.

6. Use of the chloroquine lipid according to claim 1 or 2 and / or the chloroquine lipid prepared by the method according to any one of claims 3 to 5 in the preparation of lipid nanoparticles.

7. A lipid nanoparticle, characterized in that, The lipid nanoparticle contains chloroquine lipid, cationic lipid, phospholipid, PEG lipid, and cholesterol; wherein, the chloroquine lipid is the chloroquine lipid according to claim 1 or 2 and / or the chloroquine lipid prepared by the method according to any one of claims 3 to 5.

8. The lipid nanoparticle according to claim 7, wherein, The molar ratio of the chloroquine lipid, cationic lipid, phospholipid, PEG lipid, and cholesterol is (2 - 15):(15 - 30):(3 - 10):(20 - 30):1, preferably (3 - 15):(20 - 30):(3 - 10):(22 - 28):1; Preferably, the molar ratio of the chloroquine lipid to the cationic lipid is 0.1 - 0.4:

1.

9. The lipid nanoparticle according to claim 7 or 8, wherein The cationic lipid is selected from at least one of SM - 102, MC3, and ALC - 0315, the phospholipid is DSPC, and the PEG lipid is DMG - PEG2000 and / or ALC - 0159.

10. Use of the lipid nanoparticle according to any one of claims 7 to 9 in the delivery of gene drugs.

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