Nanostructure liposome and preparation method thereof

Abstract

The invention discloses a nano-structure liposome and a preparation method thereof, and belongs to the technical field of liposome. The preparation method of the nanostructured liposome comprises the following steps: taking an oil phase and a water phase as raw materials to react to obtain the nanostructured liposome, the oil phase is prepared from glycerin monostearate, medium chain triglyceride, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacine B and cell-penetrating peptide TAT; the water phase comprises a glucose solution and Tween 80. The cucurbitacine B is applied to the aspect of anti-cancer chemotherapy drugs, solves the technical problem of poor bioavailability of the existing cucurbitacine B, and has the characteristics of good biocompatibility, low toxicity and good absorbability.

Description

Technical Field

[0001] The invention belongs to the technical field of liposomes, and in particular relates to a nanostructured liposome and a preparation method thereof. Background Art

[0002] Chemotherapy is a key method for treating tumors, but chemotherapy drugs are often accompanied by high toxicity and non-specific pharmacological activity, which not only attack tumor cells but also seriously damage normal tissues. Since chemotherapy drugs have poor targeting to tumor cells, while killing tumor cells, they also damage other normal tissues and cells in the body. In order to improve the efficacy of anticancer drugs and reduce toxic side effects, in addition to seeking breakthroughs in chemical structure, it is also necessary to innovate in dosage form design and administration methods, and constantly explore new dosage forms and optimal administration routes for anticancer drugs, in order to achieve new breakthroughs in anticancer chemotherapy drugs.

[0003] Cucurbitacin B (CuB) is a tetracyclic triterpenoid compound isolated from plants such as the Cucurbitaceae family. It is the most abundant member of the cucurbitacin family and has a wide range of pharmacological activities. CuB effectively inhibits the cell cycle of triple-negative breast cancer and induces apoptosis by precisely regulating key signaling pathways such as JAK / STAT3 and MAPK. At the same time, it can also interfere with the growth of cancer cells by inhibiting microtubule polymerization. In recent years, domestic and foreign scholars have conducted in-depth and systematic explorations of the anti-tumor potential of CuB, revealing its broad prospects in the treatment of triple-negative breast cancer.

[0004] Currently, the main cucurbitacin B preparation on the market is in the form of tablets. Cucurbitacin B is highly lipid-soluble, poorly soluble in water, and has poor oral properties. In addition, the correlation between the dosage form of cucurbitacin B and its pharmacological activity is currently unclear, making it difficult to accurately regulate the release and absorption of the drug in the body, which in turn affects the effective control of toxic and side effects. To improve this, future research may need to explore the physical and chemical properties of cucurbitacin B in more depth, optimize its dosage form design, and clarify the specific connection between it and pharmacological activity, so as to minimize toxic and side effects while ensuring the therapeutic effect. Summary of the invention

[0005] In view of the shortcomings of the prior art, the technical problem to be solved by the present invention is to solve the technical problem of poor bioavailability of cucurbitacin B, and to propose a nanostructured liposome with good biocompatibility, low toxicity and good absorbability and a preparation method thereof.

[0006] In order to solve the technical problem, the technical solution adopted by the present invention is: The present invention provides a method for preparing nanostructured liposomes, comprising reacting an oil phase and an aqueous phase as raw materials to obtain nanostructured liposomes; the oil phase comprises glyceryl monostearate, medium-chain triglycerides, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacin B, and cell-penetrating peptide TAT; and the aqueous phase comprises a glucose solution and Tween 80.

[0007] Preferably, the oil phase consists of the glyceryl monostearate, the medium-chain triglyceride, the polyethylene glycol stearate, the polyoxyethylene 35 castor oil, the cucurbitacin B, and the cell-penetrating peptide TAT; and the aqueous phase consists of the glucose solution and the Tween 80.

[0008] Preferably, the volume ratio of the oil phase to the water phase is 9:2.

[0009] Preferably, in parts by weight, the amount of the glyceryl monostearate is any value in 22-100 parts, the amount of the medium-chain triglycerides is any value in 17-100 parts, the amount of the polyethylene glycol stearate is any value in 18.7-100 parts, the amount of the polyoxyethylene 35 castor oil is any value in 20.3-100 parts, the amount of the cucurbitacin B is any value in 2-100 parts, and the amount of the cell-penetrating peptide TAT is any value in 20-100 parts.

[0010] Preferably, the aqueous phase is prepared by adding 10% Tween 80 to a 5% glucose solution.

[0011] Preferably, the cell-penetrating peptide TAT is derived from the transcription activator of HIV virus.

[0012] Preferably, the method comprises the following steps: Take the glyceryl monostearate, the medium-chain triglyceride, the polyethylene glycol stearate, the polyoxyethylene 35 castor oil, the cucurbitacin B, and the cell-penetrating peptide TAT, stir and heat in a constant temperature water bath, and use them as the oil phase for later use; Take a 5% glucose solution by mass, add 10% Tween 80 surfactant as the water phase, and heat it to the same temperature as the oil phase; Under magnetic stirring, the water phase was dropped into the oil phase for emulsification and solidified in an ice bath; The solidified mixture was transferred to a centrifuge tube and sonicated using an ultrasonic cell disruptor; After ultrasound, the water-oil mixture is filtered through a microporous filter membrane to remove impurities, thereby obtaining a TAT peptide-modified cucurbitacin B nanostructured lipid carrier.

[0013] Preferably, specifically including: Take the glyceryl monostearate, the medium-chain triglyceride, the polyethylene glycol stearate, the polyoxyethylene 35 castor oil, the cucurbitacin B, and the cell-penetrating peptide TAT, stir and heat in a constant temperature water bath at 80° C., and use them as the oil phase for later use; Take a 5% glucose solution by mass, add 10% Tween 80 surfactant as the water phase, and heat it to the same temperature as the oil phase; Under magnetic stirring, the water phase was dropped into the oil phase for emulsification for 5 min and solidified in an ice bath for 10 min; The solidified mixture was transferred to a centrifuge tube and sonicated at 35% power for 10 min using an ultrasonic cell disruptor; After ultrasound, the water-oil mixture was filtered through a 0.22 μm microporous filter membrane to remove impurities, thereby obtaining a TAT peptide-modified cucurbitacin B nanostructured lipid carrier.

[0014] Another aspect of the present invention provides nanostructured liposomes prepared by the method for preparing nanostructured liposomes described in any of the above technical solutions.

[0015] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a method for preparing nanostructured liposomes. The prepared nanostructured liposomes have the characteristics of good water solubility, being conducive to in vivo absorption, having a long half-life, and having high bioavailability, and can significantly improve the stability of cucurbitacin B; specifically: According to the charge, hydrophilicity, biocompatibility and stability of CuB, lipid materials such as monostearate, medium-chain triglycerides and polyoxyethylene 35 castor oil were selected to encapsulate CuB into a phospholipid complex, which solved the problem of poor water solubility of CuB and difficulty in in vivo absorption. The surface of CuB phospholipid complex is modified with polyethylene glycol (PEG) by using polyethylene glycol stearate to prepare long-circulating liposomes, which can reduce its nonspecific binding with plasma proteins, prolong its circulation time in the body, and reduce the risk of being recognized and cleared by the reticuloendothelial system. Long-circulating liposomes can prevent many different components in the blood, especially opsonins, from binding to them due to the presence of hydrophilic groups, thereby reducing the affinity with the mononuclear phagocyte system MPS, and can stably exist in the circulatory system and prolong its half-life, thereby increasing its uptake by tumor tissues. In addition, due to the increased permeability of capillaries caused by lesions in cancer growth sites and infection and inflammation sites, long-circulating liposomes containing drugs can increase the amount of drugs accumulated in these sites. In addition, due to the sustained release of liposome drugs directly acting on the lesion site, the therapeutic effect is enhanced, thereby solving the problem of short half-life. CuB acts on the whole body after injection, and has great toxicity and adverse reactions, which seriously limit its clinical application. The method provided by the present invention encapsulates CuB into nanostructured liposomes, and connects the cell-penetrating peptide TAT on the surface of the liposome to achieve efficient entry into cells. TAT (AYGRKKRRQRRR) is a cell-penetrating peptide derived from the transcription activator of HIV virus, which can efficiently deliver drugs into cells. It can help the carried drugs pass through the cell membrane and enter the interior of the cell, increase the efficiency of the drug on the intracellular target, and solve the problem of poor absorbability and difficulty in absorption in the body. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The morphology of nanostructured liposomes under scanning electron microscopy, where A: blank nanostructured liposomes (NLCs); B: cucurbitacin B nanostructured liposomes (CuB-NLCs); C: cucurbitacin B nanostructured liposomes with TAT (TAT-CuB-NLCs); Figure 2 is the release curve of CuB-NLCs and TAT-CuB-NLCs within 9 h (n=3); Figure 3 is the standard curve of Cou-6 solution (n=3); Figure 4 is the molecular docking diagram; Figure 5 The scratch morphology of cells in each group at different time points, including AD. Cell scratches of blank control group at 0, 12, 24, and 48 h; EH. Cell scratches of CuB group at 0, 12, 24, and 48 h; IL. Cell scratches of TAT-CuB-NLCs group at 0, 12, 24, and 48 h; Figure 6 Cytotoxicity of different concentrations of CuB and TAT-CuB-NLCs on MDA-MB-231 cells, **P<0.01, ***P<0.001 vs 0.56 μg•mL -1 CuB; **P<0.01, ***P<0.001 vs 0.56 μg·mL -1 TAT-CuB-NLC. DETAILED DESCRIPTION

[0017] The technical scheme in the specific embodiment of the present invention is described in detail and completely below. Obviously, the described embodiment is only a part of the specific implementation of the overall technical scheme of the present invention, rather than all implementations. Based on the overall concept of the present invention, all other embodiments obtained by ordinary technicians in this field fall within the scope of protection of the present invention.

[0018] The present invention provides a method for preparing nanostructured liposomes, comprising reacting an oil phase and an aqueous phase as raw materials to obtain the nanostructured liposomes; the oil phase comprises glyceryl monostearate, medium-chain triglycerides, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacin B, and transmembrane peptide TAT; and the aqueous phase comprises a glucose solution and Tween 80.

[0019] In the above method, according to the charge and hydrophilicity, biocompatibility and stability of CuB, lipid materials such as monostearate, medium-chain triglycerides, polyoxyethylene 35 castor oil are selected to encapsulate CuB into a phospholipid complex, thereby solving the problem of poor water solubility of CuB and difficult absorption in the body; polyethylene glycol stearate is used to modify the surface of the CuB phospholipid complex with polyethylene glycol (PEG) to prepare long-circulating liposomes, thereby reducing its nonspecific binding with plasma proteins, prolonging its circulation time in the body, and reducing the risk of being recognized and cleared by the reticuloendothelial system. Long-circulating liposomes can prevent many different components in the blood, especially opsonins, from binding to them due to the presence of hydrophilic groups, thereby reducing the affinity with the mononuclear phagocyte system MPS, and can stably exist in the circulatory system and prolong its half-life, thereby increasing its uptake by tumor tissues, and also due to cancer growth. The lesions at the sites of infection and inflammation cause the permeability of capillaries to increase. The long-circulating liposomes containing drugs can increase the aggregation of drugs at these sites. Since the sustained release of liposome drugs directly acts on the lesions, the therapeutic effect is enhanced, thereby solving the problem of short half-life. CuB acts on the whole body after injection, and has great toxicity and adverse reactions, which seriously limit its clinical application. The method provided by the present invention encapsulates CuB into nanostructured liposomes, and connects the membrane-penetrating peptide TAT on the surface of the liposome to achieve efficient entry into cells. TAT (AYGRKKRRQRRR) is a cell-penetrating peptide derived from the transcription activator of HIV virus, which can efficiently deliver drugs into cells. It can help the carried drugs to cross the cell membrane and enter the interior of the cell, increase the efficiency of the drug on the intracellular target, and solve the problems of poor absorbability and difficulty in absorption in the body.

[0020] In a preferred embodiment, the oil phase is composed of glyceryl monostearate, medium chain triglycerides, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacin B, and cell-penetrating peptide TAT; the aqueous phase is composed of glucose solution and Tween 80. Further, the volume ratio of the oil phase to the aqueous phase is 9:2.

[0021] In a preferred embodiment, by weight, the amount of glyceryl monostearate is any value in 22-100 parts, the amount of medium-chain triglycerides is any value in 17-100 parts, the amount of polyethylene glycol stearate is any value in 18.7-100 parts, the amount of polyoxyethylene 35 castor oil is any value in 20.3-100 parts, the amount of cucurbitacin B is any value in 2-100 parts, and the amount of cell-penetrating peptide TAT is any value in 20-100 parts; the aqueous phase is prepared by adding 10% Tween 80 to a 5% glucose solution. It is understood that the amount of glyceryl monostearate (GMS) can also be 30, 40, 50, 60, 70, 80, 90 parts and any point value within the range, the amount of medium chain triglycerides (MCT) can also be 20, 30, 40, 50, 60, 70, 80, 90 parts and any point value within the range, the amount of polyethylene glycol stearate (Solutol HS15) can also be 20, 30, 40, 50, 60, 70, 80, 90 parts and any point value within the range, polyoxyethylene 35 castor oil (Kol-liphor The amount of EL) can also be 30, 40, 50, 60, 70, 80, 90 parts and any point value within the range thereof, the amount of cucurbitacin B (CuB) can also be 10, 20, 30, 40, 50, 60, 70, 80, 90 parts and any point value within the range thereof, and the amount of the cell-penetrating peptide TAT can also be 30, 40, 50, 60, 70, 80, 90 parts and any point value within the range thereof.

[0022] In a preferred embodiment, the cell-penetrating peptide TAT is derived from the transcriptional activator of HIV virus. TAT (AYGRKKRRQRRR) is a cell-penetrating peptide derived from the transcriptional activator of HIV virus, which can efficiently deliver drugs into cells. It can help the carried drugs to cross the cell membrane and enter the cell, increasing the efficiency of the drug's action on the intracellular target.

[0023] In a preferred embodiment, the method comprises the following steps: taking glyceryl monostearate, medium-chain triglycerides, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacin B, and transmembrane peptide TAT, stirring and heating in a constant temperature water bath, and using them as the oil phase for standby use; taking a 5% glucose solution by mass, adding 10% Tween 80 surfactant as the aqueous phase, and heating it to the same temperature as the oil phase; under magnetic stirring, dripping the aqueous phase into the oil phase for emulsification, and solidifying it in an ice bath; transferring the solidified mixture to a centrifuge tube, and using an ultrasonic cell crusher for ultrasonication; after ultrasonication, filtering the above water-oil mixture through a microporous filter membrane to remove impurities, and obtaining a TAT peptide-modified cucurbitacin B nanostructured lipid carrier. Specifically comprising: Glyceryl monostearate, medium-chain triglycerides, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacin B, and cell-penetrating peptide TAT were stirred and heated in a constant temperature water bath at 80°C as the oil phase for later use; a 5% glucose solution was added with 10% Tween 80 surfactant as the water phase and heated to the same temperature as the oil phase; under magnetic stirring, the water phase was dropped into the oil phase for emulsification for 5 min and solidified in an ice bath for 10 min; the solidified mixture was transferred to a centrifuge tube and ultrasonicated at 35% power for 10 min using an ultrasonic cell disruptor; after ultrasonication, the water-oil mixture was filtered through a 0.22 μm microporous filter membrane to remove impurities, thereby obtaining the TAT peptide-modified cucurbitacin B nanostructured lipid carrier.

[0024] Another aspect of the present invention provides a nanostructured liposome prepared by the method for preparing the nanostructured liposome of any of the above technical solutions. The nanostructured liposome is a cucurbitacin B nanostructured lipid carrier modified with the transmembrane peptide TAT, which not only has high bioavailability, but also can significantly improve the stability of cucurbitacin B. Specifically, it has the following advantages: (1) The nanoliposomes of the present invention are membrane-penetrating peptide-modified natural drug monomer nanoliposome targeted preparations and have no biological toxicity.

[0025] (2) Nanotechnology can be used to make biodegradable and biocompatible polymers as drug carriers, thereby increasing the concentration of drugs at target sites or absorption sites and changing their distribution and pharmacokinetic processes in the body.

[0026] (3) The nanoliposomes of the present invention adopt a passive drug loading method and are modified by linking a substance containing a polystyryl group to the phospholipid phosphate group to form long-circulating nanoliposomes. The liposomes are protected from being recognized and taken up by opsonins in the blood, thereby reducing the clearance rate of the liposomes and prolonging their residence time in the blood, thereby prolonging the drug action time. The liposomes can effectively reach the lesion site and improve the selection of target tissues.

[0027] (4) The present invention adopts a microporous membrane filtration sterilization method, which has mild conditions and is suitable for the sterilization of liposomes.

[0028] (5) The nanoliposomes of the present invention have significant improvements in terms of encapsulation efficiency and stability compared to existing preparations.

[0029] (6) The anticancer effect of the nanoliposomes of the present invention is significantly higher than that of the existing cucurbitacin B preparations on the market.

[0030] (7) The nanoliposomes of the present invention can significantly reduce the toxicity of cucurbitacin B to cells.

[0031] The above-mentioned nanostructured liposomes also have the following characteristics: (1) Reduce drug side effects The drug concentration of ordinary cucurbitacin B preparations is high, which may lead to excessive drug concentration in non-target tissues or cells, thereby increasing the risk of side effects. The effect of cucurbitacin B nanostructured liposomes modified with the cell-penetrating peptide TAT with the same cucurbitacin B content is significantly higher than that of ordinary cucurbitacin B preparations, so the concentration of cucurbitacin B can be reduced, thereby reducing side effects.

[0032] (2) Release characteristics The drug release rate of common cucurbitacin B preparations is relatively fast, and peak concentration may be reached in a short time, and then rapidly decline. This may result in a shorter duration of action of the drug in vivo, requiring frequent administration. The cell-penetrating peptide TAT-modified cucurbitacin B nanostructured liposomes of the present invention have a sustained release effect, can slowly release drugs, delay renal excretion and metabolism, thereby extending the duration of drug action. This property makes liposomes perform well in treatments that require continuous administration.

[0033] (3) Improving drug stability The cell-penetrating peptide TAT modified cucurbitacin B nanostructured liposomes can significantly improve the stability of the drug. After being encapsulated by liposomes, the stability of the main drug can be significantly improved, thereby extending the shelf life of the drug and improving the efficacy.

[0034] (4) Bioavailability and absorbability In the nanostructured liposomes, the liposomes have a biological membrane-like structure and are modified with the cell-penetrating peptide TAT, which can better interact with the cell membrane in the body, thereby improving the bioavailability and absorbability of cucurbitacin B.

[0035] In order to more clearly and in detail introduce the nanostructured liposomes and the preparation method thereof provided by the embodiments of the present invention, they will be described below in conjunction with specific embodiments.

[0036] Example 1 A TAT-modified cucurbitacin B nanostructured liposome and a preparation method thereof, which are prepared from the following raw materials: It consists of two parts: oil phase and water phase (oil phase: water phase = 9:2): (1) Oil phase: Glyceryl monostearate (GMS): 22-100 parts Medium chain triglycerides (MCT): 17-100 servings Polyethylene glycol stearate (Solutol HS15): 18.7-100 parts Kol-liphor EL: 20.3-100 parts Cucurbitacin B (CuB): 2-100 parts Cell-penetrating peptide TAT: 20-100 copies (2) Water phase: 10% Tween 80 was added to 5% glucose solution.

[0037] The preparation method of TAT-modified cucurbitacin B nanoliposome drug comprises the following steps: (1) Weigh appropriate amounts of GMS, MCT, Solutol HS15, Kolliphor EL, CuB and TAT, heat in a constant temperature water bath at 80°C with stirring, and prepare as the oil phase; (2) Add 10% Tween 80 surfactant to a 5% glucose solution as the aqueous phase and heat to the same temperature; (3) Under magnetic stirring, the water phase was dropped into the oil phase for emulsification for 5 min and then solidified in an ice bath for 10 min; (4) Transfer the solidified mixture to a 15 mL centrifuge tube and sonicate it at 35% power for 10 min using an ultrasonic cell disruptor. (5) After sonication, the water-oil mixture was filtered through a 0.22 μm microporous filter membrane in a biosafety cabinet to remove impurities, thereby obtaining TAT peptide-modified cucurbitacin B nanostructured lipid carriers (TAT-CuB-NLCs).

[0038] The product of the present invention has been tested and experimented, and has high encapsulation efficiency, stable quality, non-toxicity, and good biological activity. The specific situation is as follows: (1) Morphological structure of liposomes Take 200 μL of each diluted NLCs, CuB-NLCs and TAT-CuB-NLCs solution and drop them on a glass slide. The samples are dried at room temperature and placed under a scanning electron microscope to observe the morphology of the nanostructured liposomes. Figure 1 shown.

[0039] (2) Particle size, polydispersity index and zeta potential measurement The particle size was measured using a nanometer potential particle size analyzer, and the results are shown in Table 1.

[0040] Table 1 Characterization results of NLCs, CuB-NLCs, and TAT-CuB-NLCs Average particle size (nm) Average polydispersity index Average Zeta Potential (mV) NLCs 49.19±2.71 0.192±0.090 -21.5±0.7 CuB-NLCs 51.99±2.04 0.207±0.042 -21.1±0.4 TAT-CuB-NLCs 36.00±2.10 0.252±0.030 +24.1±0.5 (3) Content determination The peak areas of CuB standard solutions with different concentration gradients were determined by HPLC according to the chromatographic conditions in Table 2. A standard curve was drawn with the sample mass concentration ρ (x) as the abscissa and the peak area A (y) as the ordinate.

[0041] Table 2 Chromatographic conditions Conditional parameters Parameter selection Chromatographic columns Dikma C18 column (200 mm×4.6 mm, 5 μm) Mobile phase Acetonitrile-water (volume ratio 50:50) UV detection wavelength 210 nm Injection volume 20 μL Flow rate <![CDATA[1.0 mL·min -1 ]]> Column temperature 25℃ (4) Determination of encapsulation efficiency and drug loading The encapsulation efficiency and drug loading of the nanoliposomes were determined by gel filtration chromatography combined with centrifugation. The encapsulation efficiency and drug loading were calculated according to the following formulas. The average encapsulation efficiency was 92.03%, and the average drug loading was 10.71%. Where: m 总 - the total mass of the drug in the system; m 游离 - free drug mass not encapsulated into nanoparticles; m 包 -Mass of drug encapsulated in nanoparticles.

[0042] Where: m0-total mass of nanoparticles and drugs in the system; m 包 -Mass of drug encapsulated in nanoparticles.

[0043] (5) Release like Figure 2 As shown in the figure, the drug in CuB-NLCs and TAT-CuB-NLCs can be effectively released, with cumulative release of (74.81±2.97)% and (75.43±3.05)% respectively within 9 h. There is no significant difference in release. The drug release behavior of TAT-CuB-NLCs solution conforms to the Higuchi equation (R 2 =0.9283), indicating that the TAT-CuB-NLCs prepared in this study have a sustained release effect.

[0044] (6) Stability The Cou6-NLCs and TAT-Cou6-NLCs solutions were sampled at different times using the dialysis bag method, and the fluorescence intensity of the samples was measured to calculate the in vitro release rate. Figure 3 As shown, the cumulative release amounts of Cou6-NLCs and TAT-Cou6-NLCs within 2 h were (22.98±0.34)% and (20.01±0.29)%, respectively, with good stability.

[0045] (7) Molecular docking Screening of cucurbitacin B, the active ingredient of gua li, and targets for triple-negative breast cancer, such as Figure 4 As shown, molecular docking was performed and it was preliminarily determined that cucurbitacin B has anticancer activity against triple-negative breast cancer.

[0046] (8) Cell scratch assay to verify its biological activity like Figure 5 As shown, the cell-penetrating peptide TAT-modified cucurbitacin B nanostructured lipid carriers (TAT-CuB-NLCs) significantly inhibited cell migration.

[0047] (9) MTT assay to verify cell viability like Figure 6As shown, the cell activity of the cell viability of the cucurbitacin B nanostructured lipid carrier (TAT-CuB-NLCs) group modified with the cell-penetrating peptide TAT was significantly lower than that of the cucurbitacin B (CuB) group.

Claims

1. A method for preparing nanostructured liposomes, characterized in that: The invention comprises obtaining nanostructured liposomes by reacting an oil phase and an aqueous phase as raw materials; the oil phase comprises glyceryl monostearate, medium-chain triglyceride, polyethylene glycol stearate, polyoxyethylene 35 castor oil, cucurbitacin B and cell-penetrating peptide TAT; and the aqueous phase comprises glucose solution and Tween 80.

2. The method for preparing nanostructured liposomes according to claim 1, characterized in that: The oil phase consists of the glyceryl monostearate, the medium-chain triglyceride, the polyethylene glycol stearate, the polyoxyethylene 35 castor oil, the cucurbitacin B, and the cell-penetrating peptide TAT; the water phase consists of the glucose solution and the Tween 80.

3. The method for preparing nanostructured liposomes according to claim 1 or 2, characterized in that: The volume ratio of the oil phase to the water phase is 9:

2.

4. The method for preparing nanostructured liposomes according to claim 1 or 2, characterized in that: In parts by weight, the amount of the glyceryl monostearate is any value in the range of 22-100 parts, the amount of the medium-chain triglyceride is any value in the range of 17-100 parts, the amount of the polyethylene glycol stearate is any value in the range of 18.7-100 parts, the amount of the polyoxyethylene 35 castor oil is any value in the range of 20.3-100 parts, the amount of the cucurbitacin B is any value in the range of 2-100 parts, and the amount of the cell-penetrating peptide TAT is any value in the range of 20-100 parts.

5. The method for preparing nanostructured liposomes according to claim 1 or 2, characterized in that: The aqueous phase is prepared by adding 10% Tween 80 to a 5% glucose solution.

6. The method for preparing nanostructured liposomes according to claim 1, characterized in that: The cell-penetrating peptide TAT is derived from the transcription activator of HIV virus.

7. The method for preparing nanostructured liposomes according to claim 1, characterized in that: The following steps are involved: Take the glyceryl monostearate, the medium-chain triglyceride, the polyethylene glycol stearate, the polyoxyethylene 35 castor oil, the cucurbitacin B, and the cell-penetrating peptide TAT, stir and heat in a constant temperature water bath, and use them as the oil phase for later use; Take a 5% glucose solution by mass, add 10% Tween 80 surfactant as the water phase, and heat it to the same temperature as the oil phase; Under magnetic stirring, the water phase was dropped into the oil phase for emulsification and solidified in an ice bath; The solidified mixture was transferred to a centrifuge tube and sonicated using an ultrasonic cell disruptor; After ultrasound, the water-oil mixture is filtered through a microporous filter membrane to remove impurities, thereby obtaining a TAT peptide-modified cucurbitacin B nanostructured lipid carrier.

8. The method for preparing nanostructured liposomes according to claim 7, characterized in that: Specifically include: Take the glyceryl monostearate, the medium-chain triglyceride, the polyethylene glycol stearate, the polyoxyethylene 35 castor oil, the cucurbitacin B, and the cell-penetrating peptide TAT, stir and heat in a constant temperature water bath at 80° C., and use them as the oil phase for later use; Take a 5% glucose solution by mass, add 10% Tween 80 surfactant as the water phase, and heat it to the same temperature as the oil phase; Under magnetic stirring, the water phase was dropped into the oil phase for emulsification for 5 min and solidified in an ice bath for 10 min; The solidified mixture was transferred to a centrifuge tube and sonicated at 35% power for 10 min using an ultrasonic cell disruptor; After ultrasound, the water-oil mixture was filtered through a 0.22 μm microporous filter membrane to remove impurities, thereby obtaining a TAT peptide-modified cucurbitacin B nanostructured lipid carrier.

9. The nanostructured liposome prepared according to the method for preparing the nanostructured liposome according to any one of claims 1 to 8.

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