Lymphatic targeting carrier and preparation method and application thereof

Abstract

The invention discloses a lymphatic targeting carrier as well as a preparation method and application thereof, and belongs to the technical field of biomedical materials. The lymphatic targeting carrier is composed of nanocrystals wrapped by lipidosome and hyaluronidase, the liposome is composed of DSPC, Chol, PS and MTO (Methanol To Olefin). The nanocrystal comprises any one of a medicine and an active substance. According to the invention, ivermectin is used as a model drug to prepare the PS / MTO co-modified IVM-NCs, and the PS / MTO co-modified IVM-NCs and hyaluronidase are delivered together, so that the lymphatic accumulation of the drug is enhanced. The nano-drug is directly delivered into the skin by bypassing a cuticle barrier through subcutaneous injection, hyaluronidase is used for degrading a skin extracellular matrix barrier, increasing skin diffusion of nano-crystal particles, improving the ability of the nano-crystal particles to enter a lymphatic vasculature and improving the drug delivery efficiency and the drug delivery accuracy; and a new strategy is provided for lymphatic targeted delivery of fat-soluble drugs.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedical materials, and in particular relates to a lymphatic targeting transfersome and a preparation method and application thereof. Background Art

[0002] In recent years, the lymphatic system has been found to be closely related to acute and critical diseases such as cancer metastasis, cardiovascular atherosclerosis and heart failure, infectious diseases (such as lymphatic filariasis, HIV, hepatitis and Ebola virus, etc.), diabetes, lymphedema, schizophrenia and tuberculosis. The lymphatic system is an important defense system in the human body that promotes the mixing of immune cells and thus regulates adaptive immune responses. The lymphatic vasculature is composed of a single layer of lymphatic endothelial cells (LECs), which loosely overlap each other to form button-like connections. These button-like connections form flaps between LECs, creating pores with a diameter of about 2-3 μm, which can accommodate fluids, macromolecules and immune cells to enter the capillary lymphatic lumen to form lymph.

[0003] The unique physiological structure of lymph makes it very suitable for the delivery of nanoparticles (NPs). Currently, NPs for lymphatic-directed therapy, such as the Gardasil® / Cervarix® vaccine based on virus-like protein nanoparticles and the Comirnaty® vaccine based on mRNA and lipid NPs, have been approved by the U.S. Food and Drug Administration (FDA) for the prevention of human papillomavirus infection and COVID-19, respectively.

[0004] Most of these nanoparticles are captured by lymphatic capillaries or antigen-presenting cells (such as macrophages and dendritic cells in peripheral tissues) through subcutaneous, intradermal or intramuscular injections and enter the lymphatic circulation. Compared with oral delivery (limited by the complex gastrointestinal environment of the human body and the barrier of tightly connected epithelial cells) and intranodal injection (hindered by the technical challenges of locating lymph nodes), nanoparticles have irreplaceable advantages in lymphatic targeting through subcutaneous, intradermal or intramuscular injections. However, nanoparticles also face many problems, such as the use of a large amount of carrier materials resulting in low drug loading. For example, patent CN113577300A discloses a targeted liposome drug delivery system, whose drug loading is only 3-6% by mass, which needs to be further improved; the lymphatic targeting efficiency is low, most of the nanoparticles remain in the peripheral injection site, and the absorption of capillaries will also reduce the lymphatic targeting efficiency.

[0005] Therefore, there is an urgent need for a delivery system with high drug loading and efficient lymphatic targeting. Summary of the invention

[0006] The purpose of the present invention is to provide a lymphatic targeting delivery body and its preparation method and application. The lymphatic targeting delivery body provided by the present invention can break through the skin extracellular matrix (ECM) barrier and actively realize lymphatic delivery of fat-soluble drugs during drug delivery.

[0007] In order to achieve the above object, the technical solution of the present invention is: In a first aspect of the present invention, a lymphatic targeting delivery body is provided, which is composed of liposome-encapsulated nanocrystals and hyaluronidase; The liposomes are composed of distearoylphosphatidylcholine (DSPC), cholesterol (Chol), phosphatidylserine (PS) and mitoxantrone hydrochloride (MTO); The nanocrystals include any one of a drug and an active substance.

[0008] The present invention uses the substance to be delivered as the core, DSPC and Chol as liposomes to encapsulate the core, PS and MTO are modified to the core through the liposome shell, and are delivered together with hyaluronidase, thereby enhancing the accumulation of drugs or active substances in the lymph.

[0009] It should be noted that the administration method of the lymphatic targeted delivery body of the present invention is subcutaneous injection, which bypasses the stratum corneum barrier and directly delivers the nanodrug into the skin; secondly, hyaluronidase is used to degrade the skin ECM barrier, increase the skin diffusion of liposome-encapsulated nanocrystals, and improve their ability to enter the lymphatic vascular system; finally, liposome-encapsulated nanocrystals modified with active targeting ligands are used for lymphatic targeting, providing a new strategy for lymphatic delivery of drugs, especially fat-soluble drugs.

[0010] In some embodiments of the invention, the nanocrystals include a lipid-soluble drug.

[0011] In some embodiments of the present invention, the nanocrystals further include a stabilizer, and the stabilizer is selected from one of vitamin E polyethylene glycol succinate (TPGS) and polyether F68 (F68), preferably TPGS.

[0012] In some embodiments of the present invention, the nanocrystals are prepared by mixing the fat-soluble drug and TPGS in a mass ratio of 0.9-1.1:0.9-1.1. The mass ratio is preferably 1:1. At this ratio, the obtained nanocrystal particles have a particle size of less than 100 nm.

[0013] In some embodiments of the present invention, the molar ratio of DSPC, Chol, PS and MTO is 6:4:2-3:1-3, preferably 6:4:2.5:2.

[0014] In some embodiments of the present invention, the mass ratio of the liposome-encapsulated nanocrystals to hyaluronidase is 1:1-2, preferably 1:1.5.

[0015] In some embodiments of the present invention, the particle size of the liposome-encapsulated nanocrystals is 150-200 nm, preferably 160-190 nm; The drug loading in the liposome-encapsulated nanocrystals is 15% to 25%, preferably 19% to 22%, by mass percentage; the encapsulation rate of the nanocrystals is 50.00% to 99.99%.

[0016] The second aspect of the present invention provides a method for preparing the above-mentioned lymphatic targeting transfersome, comprising: Under ultrasonic conditions, a stabilizer is added to a transfersome solution to obtain a nanocrystal solution; the transfersome is any one of a drug and an active substance; DSPC, Chol, PS and MTO were dissolved in a volatile solvent and distilled under reduced pressure to form a film to obtain liposomes; The nanocrystal solution is added into the liposome, reacted, ultrasonicated, centrifuged and concentrated to obtain the nanocrystal solution encapsulated by the liposome.

[0017] The invention prepares a nano crystal solution by mixing the transfer body with a stabilizer, which has high stability and greatly improves the drug loading rate of the lymphatic targeting transfer body.

[0018] In some embodiments of the present invention, the mass ratio of the transfersome to the stabilizer is 0.9-1.1:0.9-1.1, preferably 1:1.

[0019] Preferably, the transfersome is a fat-soluble drug, and the transfersome solution is an alcohol solution.

[0020] In some embodiments of the present invention, the mass ratio of the total mass of DSPC, Chol, PS and MTO to the mass of the transfer body is 7-9:1, preferably 8:1.

[0021] In some embodiments of the present invention, the concentration of the transfersome in the liposome-encapsulated nanocrystal solution is 5-15 mg / mL, preferably 10 mg / mL.

[0022] The third aspect of the present invention provides a use of the above-mentioned lymphatic targeting transfersome or the lymphatic targeting transfersome prepared by the above-mentioned preparation method in the preparation of a drug for preventing and / or treating lymph-related diseases.

[0023] It is understandable that the above-mentioned lymphatic targeting delivery body or the lymphatic targeting delivery body prepared by the above-mentioned preparation method can be used in the field of drug delivery, for delivering drugs or active substances into cells or animals, and this application does not include application in treatment methods or diagnosis methods for diseases.

[0024] The lymphatic targeting delivery body provided by the present invention can deliver drugs or active substances to the lymph in a targeted manner, and therefore, it can also be used to prepare drugs for lymphatic-related diseases.

[0025] Preferably, the lymph nodes include any one or more of popliteal lymph nodes, sacroiliac lymph nodes, renal lymph nodes and inguinal lymph nodes. As the administration time increases, the lymph-targeted transfersome is gradually delivered from the popliteal lymph nodes (primary lymph nodes) to the sacroiliac lymph nodes (secondary lymph nodes), renal lymph nodes (tertiary lymph nodes) and inguinal lymph nodes, while the axillary lymph nodes are not delivered. Therefore, the lymph nodes do not include the axillary lymph nodes.

[0026] The beneficial effects of the present invention are: The present invention provides a lymphatic targeting delivery body, which can overcome the skin barrier and the skin ECM barrier, actively realize the lymphatic delivery of fat-soluble drugs, and has broad application prospects in the field of drug delivery.

[0027] The lymphatic targeting transfer body of the present invention bypasses the stratum corneum barrier by subcutaneous injection, directly delivers the nano drug into the skin, and the co-delivered hyaluronidase is used to degrade the ECM barrier, increase the skin diffusion of the liposome-encapsulated nanocrystal particles, and improve their ability to enter the lymphatic vascular system, thereby greatly improving the drug administration efficiency and accuracy, and enhancing the accumulation of drugs in the lymph. The ligand-modified liposome-encapsulated nanocrystal particles with active targeting, wherein PS and MTO are modified to the nanocrystal particle core through the liposome shell, are used for lymphatic targeting, and have high lymphatic targeting. The nanocrystal particles have high stability, and greatly improve the drug loading rate of the lymphatic targeting transfer body.

[0028] The lymphatic targeting transfersome provided by the present invention has simple preparation process conditions, stable storage, cost saving, is suitable for mass production, and can be widely used in the field of drug delivery. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] The accompanying drawings in the specification, which constitute a part of the present invention, are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0030] Figure 1 This is the particle size curve of IVM-NCs obtained by using different stabilizers in Example 1 of the present invention; Figure 2The IVM-NCs suspension obtained by using different stabilizers in Example 1 of the present invention; Figure 3 This is a screening diagram of the ratio of stabilizer to drug when F68 is a stabilizer in Example 1 of the present invention; Figure 4 This is a screening diagram of the ratio of stabilizer to drug when TPGS is used as a stabilizer in Example 1 of the present invention; Figure 5 This is a stability curve of IVM-NCs when TPGS (A) or F68 (B) is a stabilizer in Example 1 of the present invention; Figure 6 : is the particle size curve of IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO in Example 1 of the present invention; Figure 7 TEM images of IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO in Example 1 of the present invention; Figure 8 This is a stability curve diagram of IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO in Example 1 of the present invention; Fig. 9 DSC graphs of IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO in Example 1 of the present invention; Fig.10 The staining effect of lymph nodes of rats in the saline group at different times in Example 1 of the present invention; Fig.11 The staining results of rat lymph nodes at different times in the L-IVM-PS-MTO group in Example 1 of the present invention are shown; Fig.12 The staining effect of rat lymph nodes at different times in the H+L-IVM-PS-MTO group in Example 1 of the present invention; Fig.13 The graphs show the changes in MTO content in rat lymph nodes at different times in the L-IVM-PS-MTO solution group (a) and the H+L-IVM-PS-MTO group (b) in Example 1 of the present invention. DETAILED DESCRIPTION

[0031] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0032] The raw materials and reagents used in the following examples are all commercially available.

[0033] Example 1 A method for preparing a lymphatic targeting transfersome comprises the following steps: 1. Materials and reagents: Polyether F68 (F68), Distearoyl Phosphatidylcholine (DSPC), Hydroxypropyl Methylcellulose (HPMC), Polyvinylpyrrolidone K30 (PVP K30), Phosphatidylserine (PS), Mitoxantrone Hydrochloride (MTO), Vitamin E Polyethylene Glycol Succinate (TPGS), Ivermectin (IVM), Chloroform, Anhydrous Ethanol, Hyaluronidase (HAase) 2. Instruments and Equipment: Nano® Zetasizer (Malvern Instruments, Malvern, UK), transmission electron microscope, DiamondTG / DTA (Perkin Elmer, Waltham, USA), confocal Raman spectrometer, and high performance liquid chromatography.

[0034] 3. Test methods 1. Preparation and optimization of ivermectin nanocrystals (IVM-NCs) 1.1 Experimental methods: 20 mg / mL IVM ethanol solution, four stabilizers, TPGS, F68, HPMC, and PVP K30, were selected respectively, and the relevant conditions were: stabilizer: drug mass ratio was 2:1; organic phase: aqueous phase (i.e., IVM ethanol solution) = 0.25:4.75; the organic phase was injected into the aqueous phase under the conditions of ultrasonic power of 45%, 3 on / 2 off / 5 min to obtain IVM-NCs solution.

[0035] 1.2 Condition optimization: ① Stability study: Measure stability at 1, 2, 5, 10, 15, and 20 days to observe particle size and polydispersity index (PDI) ②Stabilizer / drug ratio screening: IVM:F68=1:1, 1:2, 2:1 and IVM:TPGS=5:1, 2:1, 1:1, 1:2, mass ratio.

[0036] 1.3 Experimental results: ①Stabilizer screening results: Depend on Figure 1 , Figure 2 As shown in Table 1, when the stabilizers are TPGS and F68, the IVM-NCs solution is clear and the particle size is <100 nm. TPGS and F68 can be used as stabilizers for IVM.

[0037] Table 1 Screening of stabilizer types

[0038] ②Stabilizer / drug ratio screening: Depend on Figure 3 , Figure 4 It can be seen from Table 2 and Table 3 that when IVM:F68=2:1, the particle size of IVM-NCs is <100 nm, and as the stabilizer ratio decreases, the particle size of IVM-NCs gradually increases. When F68 is used as a stabilizer, the particle size is better when IVM:F68=2:1. When TPGS is used as a stabilizer, the particle size of IVM-NCs is <100 nm when the IVM:TPGS ratio is 2:1, 1:1, and 1:2, and the particle size is the smallest when the ratio is 1:1.

[0039] Table 2 F68 as stabilizer drug ratio screening

[0040] Table 3 Screening of drug ratios using TPGS as stabilizer

[0041] ③Stability inspection: like Figure 5 As shown, after stabilization with TPGS and F68, the particle size of IVM-NCs changed uniformly within 20 days and the stability was good.

[0042] 1.4 Experimental conclusion: Finally, considering the particle size, PDI and subsequent drug loading issues, TPGS was selected as the stabilizer, and the mass ratio of IVM to TPGS was 1:1 to prepare IVM-NCs.

[0043] The IVM-NCs used in the following experiments were prepared by the following method: 20 mg / mL IVM ethanol solution, TPGS was used as a stabilizer, and the relevant conditions were: the mass ratio of stabilizer:drug was 1:1; organic phase: aqueous phase (i.e., IVM ethanol solution) = 0.25:4.75; the organic phase was injected into the aqueous phase under the condition of 3 on / 2 off / 5 min to obtain IVM-NCs solution.

[0044] 2. Preparation and morphological observation of liposome-encapsulated IVM-NCs (L-IVM) 2.1 L-IVM preparation Experimental methods: Preparation of L-IVM-PS: DSPC, Chol and PS were added to chloroform in a molar ratio of 6:4:2.5, and the film was formed by vacuum distillation at 55°C. 5 mL of IVM-NCs solution was added, hydrated at 55°C for 30 min, ultrasonicated for 3 min, and centrifuged at 4000 rpm for 20 min to obtain L-IVM-PS solution. The prepared L-IVM-PS solution was centrifuged at 4000 rpm for 20 min using an ultrafiltration tube to prepare L-IVM-PS concentrate.

[0045] Preparation of L-IVM-PS-MTO: DSPC, Chol, PS and MTO were taken according to the molar ratio of DSPC:Chol:PS:MTO of 6:4:2.5:0.2, added into chloroform, and distilled under reduced pressure at 55 °C to form a film. 5 mL of IVM-NCs solution was added, hydrated at 55 °C for 30 min, ultrasonicated for 3 min, and centrifuged at 4000 rpm for 20 min to obtain L-IVM-PS-MTO solution.

[0046] Finally, the prepared L-IVM-PS-MTO solution was centrifuged at 4000 rpm for 20 min using an ultrafiltration tube to prepare a L-IVM-PS-MTO concentrate.

[0047] 2.2 L-IVM morphology observation Experimental methods: Nano® Zetasizer (Malvern Instruments, Malvern, UK) was used to measure the particle size, PDI, and Zeta potential of IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO.

[0048] Transmission electron microscopy was used to observe the morphology of IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO.

[0049] Experimental results: Figure 6 As shown in Table 4, the particle sizes of L-IVM-PS and L-IVM-PS-MTO were 145.90±31.05 nm and 175.30±11.50 nm, respectively. Compared with IVM-NCs, the particle sizes were slightly larger (84.80±7.72 nm). The prepared liposomes had uniform particle sizes and PDIs were all less than 0.3.

[0050] Table 4 Particle size determination results

[0051] Note: TEM and particle size analyzer have different measurement principles, so there are usually some differences. But the overall trend is the same.

[0052] Figure 7 TEM showed that IVM-NCs, L-IVM-PS, and L-IVM-PS-MTO all showed spherical morphology, and the particle size of IVM-NCs slightly increased after being encapsulated by liposomes.

[0053] 2.3 L-IVM stability observation The IVM-NCs, L-IVM-PS and L-IVM-PS-MTO in 2.2 were stored at 4 °C for 75 days, and the particle size and PDI of IVM-NCs, L-IVM-PS and L-IVM-PS-MTO were measured using Nano® Zetasizer (Malvern Instruments, Malvern, UK).

[0054] The experimental results are as follows Figure 8 shown.

[0055] pass Figure 8 It can be seen from the data that the L-IVM-PS-MTO prepared in this example has excellent storage stability, and its particle size does not change much after being stored at 4°C for 75 days.

[0056] It should be noted that it has been verified that the L-IVM-PS-MTO prepared using the parameters described in the present invention has excellent storage stability and a longer storage life. However, the L-IVM-PS-MTO prepared using values ​​outside the parameter range described in the present invention does not have similar stability and has a short storage life.

[0057] 3. Structural Characterization 3.1 Experimental methods: Diamond TG / DTA (Perkin Elmer, Waltham, USA) was used to study the thermal properties and structural characteristics of IVM raw materials, stabilizer TPGS, IVM-NCs, blank liposomes (L-PS, L-PS-MTO), L-IVM-PS, and L-IVM-PS-MTO, respectively.

[0058] The preparation method of blank liposomes L-PS was compared with that of L-IVM-PS, but 5 mL of IVM-NCs solution was not added.

[0059] The preparation method of blank liposome L-PS-MTO was compared with that of L-IVM-PS-MTO, but 5 mL of IVM-NCs solution was not added.

[0060] 3.2 Experimental results: In the DSC graph ( Fig. 9 ), IVM showed an endothermic peak at 157°C, and TPGS showed a melting point at 41°C. After the two were prepared into IVM-NCs, the endothermic peaks of TPGS and IVM disappeared, indicating that IVM-NCs were in an amorphous state; due to the large presence of lipid material DSPC, the lipid mixture showed an endothermic peak at 55°C. L-PS (blank liposomes), L-PS-MTO (blank liposomes), L-IVM-PS, and L-IVM-PS-MTO also showed endothermic peaks similar to lipids.

[0061] 4. Determination of L-IVM Encapsulation Efficiency 4.1 Experimental methods: The L-IVM-PS solution and L-IVM-PS-MTO solution before centrifugation of 2.1 L-IVM were determined by HPLC. The concentration of IVM in the supernatant was collected by centrifugation, and the encapsulation efficiency (EE%) and drug loading (DL%) of IVM were calculated using the equation.

[0062]

[0063]

[0064] Experimental results: The drug loading and encapsulation efficiency of L-IVM-PS were 21.07±4.08% and 81.27±4.38%, respectively; the drug loading and encapsulation efficiency of L-IVM-PS-MTO were 20.46±1.21% and 80.66±19.04%, respectively. The drug loading was much higher than that of traditional liposome preparations of poorly soluble drugs (usually less than 10%, only a few percent).

[0065] 5. In vivo lymphatic delivery experiment of L-IVM-PS-MTO: Adaptive feeding was performed one week before the experiment. All procedures were in accordance with the Guide for the Care and Use of Laboratory Animals. Rats were divided into three groups. Physiological saline, L-IVM-PS-MTO solution, and H+L-IVM-PS-MTO solution were injected subcutaneously into the dorsum of the rats. Three rats were dissected at 1 h, 2 h, and 4 h after administration in the saline, L-IVM-PS-MTO, and H+L-IVM-PS-MTO solution groups, and the popliteal lymph nodes, sacroiliac lymph nodes, renal lymph nodes, inguinal lymph nodes, and axillary lymph nodes were removed, photographed, and their staining was observed. The lymph nodes were taken, squeezed and washed with saline to remove blood as much as possible, and dried with filter paper. 0.55 mL of ascorbic acid-citrate buffer (0.1 mol / L, pH 3.0) was added to the accurately weighed portion of each tissue, and the mixture was homogenized.

[0066] 0.55 mL of homogenate sample was mixed with 0.10 mL of methanol, 0.10 mL of formic acid, 0.05 mL of 20% trichloroacetic acid, and 0.20 mL of chloroform in a polypropylene tube, vortexed for 3 min, centrifuged at 1600 × g for 10 min, and the supernatant was stored in a refrigerator. Before the determination, the supernatant was centrifuged again at 1600 × g for 10 min. 20 μL was used for HPLC: column C18 (250 mm × 4.5 mm inner diameter, 5 μm), mobile phase methanol-0.16 mol / L ammonium formate (48:52, pH 2.7), flow rate 1.0 mL / min, detection wavelength 658 nm.

[0067] Experimental results and analysis: pass Figure 10-13 By comparison, it can be seen that the co-delivery of hyaluronidase can enhance the accumulation of drugs in the lymph.

[0068] According to the lymph node staining time chart and the MTO content analysis in each group of lymph nodes ( Figure 10-13 ) showed that with the extension of administration time, the delivery bodies of each group were gradually delivered from the popliteal lymph nodes (primary lymph nodes) to the sacroiliac lymph nodes (secondary lymph nodes), renal lymph nodes (tertiary lymph nodes), and inguinal lymph nodes. The effect became better and better, and the staining color became darker and darker. No delivery was observed in the axillary lymph nodes in each group.

[0069] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A lymphatic targeting transfersome, characterized in that: It is composed of liposome-encapsulated nanocrystals and hyaluronidase; The liposome is composed of distearoyl phosphatidylcholine, cholesterol, phosphatidylserine and mitoxantrone hydrochloride; The nanocrystals include any one of a drug and an active substance.

2. The lymphatic targeting delivery body according to claim 1, characterized in that The nanocrystals include a lipid-soluble drug; Preferably, the nanocrystals further include a stabilizer, and the stabilizer is selected from one of vitamin E polyethylene glycol succinate and polyether F68, preferably vitamin E polyethylene glycol succinate; Preferably, the nanocrystals are prepared by mixing a fat-soluble drug and vitamin E polyethylene glycol succinate in a mass ratio of 0.9-1.1:0.9-1.

1.

3. The lymphatic targeting delivery body according to claim 1, characterized in that The molar ratio of the distearoyl phosphatidylcholine, cholesterol, phosphatidylserine and mitoxantrone hydrochloride is 6:4:2-3:1-3.

4. The lymphatic targeting delivery body according to claim 1, characterized in that The mass ratio of the nanocrystals encapsulated by the liposomes to the hyaluronidase is 1:1-2.

5. The lymphatic targeting delivery body according to any one of claims 1 to 4, characterized in that: The particle size of the nanocrystals encapsulated by the liposomes is 150-200 nm; The drug loading in the nanocrystals encapsulated by the liposomes is 15% to 25% by mass; and the encapsulation rate of the nanocrystals is 50.00% to 99.99%.

6. A method for preparing the lymphatic targeting transfersome according to claim 1, characterized in that: include: Under ultrasonic conditions, a stabilizer is added to the transfersome solution to obtain a nanocrystal solution; The transfer body is any one of a drug and an active substance; Distearoylphosphatidylcholine, cholesterol, phosphatidylserine and mitoxantrone hydrochloride are dissolved in a volatile solvent, and distilled under reduced pressure to form a film to obtain liposomes; The nanocrystal solution is added into the liposome, reacted, ultrasonicated, centrifuged and concentrated to obtain the nanocrystal solution encapsulated by the liposome.

7. The preparation method according to claim 6, characterized in that: The mass ratio of the transfer agent to the stabilizer is 0.9-1.1:0.9-1.1; Preferably, the transfersome is a fat-soluble drug, and the transfersome solution is an alcohol solution.

8. The preparation method according to claim 6, characterized in that: The mass ratio of the total mass of the distearoyl phosphatidylcholine, cholesterol, phosphatidylserine and mitoxantrone hydrochloride to the mass of the transfersome is 7-9:

1.

9. The preparation method according to claim 6, characterized in that: In the liposome-encapsulated nanocrystal solution, the concentration of the transfersome is 5-15 mg / mL.

10. Use of the lymphatic targeting transfersome according to claim 1 or the lymphatic targeting transfersome prepared by the preparation method according to any one of claims 6 to 9 in preparing a drug for preventing and / or treating lymphatic related diseases; Preferably, the lymph nodes include any one or more of popliteal lymph nodes, sacroiliac lymph nodes, perirenal lymph nodes and inguinal lymph nodes.

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