Preparation method and application of heparin-targeted dihydroartemisinin-loaded antimalarial nano-liposome freeze-dried powder medicine

Abstract

The invention belongs to the technical field of biological medicines, and discloses a preparation method of a heparin targeted dihydroartemisinin-loaded antimalarial nano-liposome freeze-dried powder drug, and the preparation method comprises the following steps: S1, synthesis of DSPE-PEG2000-heparin: coupling dalteparin and DSPE-PEG2000-NHS through an amination reaction, and carrying out dialysis purification and freeze-drying; s2, dissolving soybean lecithin, cholesterol, a DSPE-PEG2000-heparin polymer and dihydroartemisinin in 5mL of chloroform, evaporating by using a rotary evaporator under low pressure to form a film, and adding deionized water for dissolving; s3, deionized water is added for film hydration, ultrasonic treatment is conducted for 1 min in a water bath at the temperature of 20 DEG C, 100 nm film extrusion treatment is conducted, and HEP-DHA-coated Lips is obtained; and S4, adding a freeze-drying protective agent, and performing freeze-drying to obtain the HEP-DHA-coated Lips freeze-dried powder preparation. The heparin-dihydroartemisinin liposome nanoparticle freeze-dried powder medicine prepared by the invention has excellent anti-malarial performance, and the problems that the traditional dihydroartemisinin is poor in water solubility, short in half-life period and low in bioavailability are solved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine, and particularly relates to a preparation method and application of a heparin-targeted anti-malaria nano-liposome freeze-dried powder drug loaded with dihydroartemisinin. BACKGROUND

[0002] Malaria is a disease transmitted by female anopheles infected with plasmodium, and mainly prevails in tropical regions. The life history of plasmodium includes the development process of two stages in the human body and anopheles. The intraerythrocytic development of plasmodium mainly experiences four stages of ring, trophozoite, merozoite and schizont, and the repeated schizogony of the intraerythrocytic stage is the direct cause of the clinical symptoms of malaria, which may cause intermittent fever, severe anemia, hypoglycemia, respiratory stress and organ damage, including brain coma or acute kidney injury.

[0003] Functional nanomaterials prepared by nanotechnology can reduce the drug carrier to the nanometer level, enhance the solubility and stability of the encapsulated drug, and the surface can be coated with antigen-related receptors to target the delivery of drugs to the lesion. This method can overcome the influence of the microenvironment and biological barriers in vivo, and provide protection for drug delivery and release in the body. The invasion of plasmodium into red blood cells is a rapid and complex process, mainly including the adhesion of merozoite and host red blood cells, the formation of tight junction complex, the formation of plasmodium vacuole and the closure of red blood cell membrane. Many studies have shown that heparin or sulfated heparin is one of the receptors for the adhesion of invading merozoites to the surface of red blood cells. Heparin and heparin molecules have also been shown to inhibit the invasion of merozoites and destroy the “rosette” formed by plasmodium-infected red blood cells to escape the immune system of the body.

[0004] At present, artemisinin and its derivatives are the most effective drugs for treating malaria, and artemisinin combination therapy (ACT) is still the most commonly used treatment for clinical treatment of malaria. However, artemisinin and its derivatives have problems such as short half-life, limited bioavailability and increasing drug resistance, which prompts researchers to explore new strategies for the development of anti-malaria drugs, and therefore, a preparation method and application of a heparin-targeted anti-malaria nano-liposome freeze-dried powder drug loaded with dihydroartemisinin are needed. SUMMARY

[0005] The application provides a preparation method and application of a heparin-targeted anti-malaria nano-liposome freeze-dried powder drug loaded with dihydroartemisinin, and aims to solve the problems of short half-life, limited bioavailability and increasing drug resistance of artemisinin and its derivatives in the treatment of malaria by artemisinin and its derivatives in the prior art.

[0006] The first aspect is a heparin-targeted anti-malaria nano-liposome lyophilized powder drug for carrying dihydroartemisinin, which is composed of a liposome nano-drug delivery system of dihydroartemisinin, liposomes and DSPE-PEG2000-heparin, the liposome nano-drug delivery system is prepared by a conventional thin film hydration method, and the heparin is coated on the outer layer of the nanoparticles, and a lyophilization protective agent is added to obtain the HEP-DHA@Lips lyophilized powder preparation.

[0007] Further, the particle size range of the HEP-DHA@Lips lyophilized powder preparation is 125.42±3.24nm, and the absolute value of Zeta potential is more than 20mV.

[0008] Further, the drug loading capacity of dihydroartemisinin is 10.58±0.67%.

[0009] The second aspect is a preparation method of a heparin-targeted anti-malaria nano-liposome lyophilized powder drug for carrying dihydroartemisinin, which comprises the following steps: S1: synthesis of DSPE-PEG2000-heparin: coupling of heparin and DSPE-PEG2000-NHS by amination reaction, dialysis purification and lyophilization; S2: dissolving soybean lecithin, cholesterol, DSPE-PEG2000-heparin polymer and dihydroartemisinin in 5mL chloroform, evaporating into a film under low pressure, and then dissolving in deionized water; S3: adding deionized water for thin film hydration, and ultrasonic treatment in a 20℃ water bath for 1min, and extruding through a 100nm membrane to obtain HEP-DHA@Lips; S4: adding a lyophilization protective agent, and lyophilizing to obtain the HEP-DHA@Lips lyophilized powder preparation.

[0010] Further, after obtaining the HEP-DHA@Lips in S3, it needs to be dialyzed in deionized water for 4h to remove the unsuccessfully loaded DSPE-PEG2K-heparin and dihydroartemisinin.

[0011] Further, the mass ratio of HEP-DHA@Lips to lyophilization protective agent in S4 is 1:5.

[0012] Further, the preparation comprises soybean lecithin, cholesterol, DSPE-PEG2000-heparin polymer and dihydroartemisinin, and the molar ratio of each component is 10:2:1:1.

[0013] The third aspect is the use of a heparin-targeted anti-malaria nano-liposome lyophilized powder drug for carrying dihydroartemisinin, which is used for inhibiting the proliferation of malaria parasites in blood and prolonging the survival time of infected hosts.

[0014] Compared with the prior art, the present application has at least the following beneficial effects: Based on further analysis and research on the problems of the prior art, the prepared heparin-dihydroartemisinin liposome nanoparticle freeze-dried powder drug has excellent antimalarial performance, which improves the problems of poor water solubility, short half-life, low bioavailability and other problems of traditional dihydroartemisinin. The heparin-dihydroartemisinin liposome nanoparticle utilizes the targeting of heparin to malignant malaria schizonts or infected red blood cells, can specifically bind to malignant malaria schizonts or infected red blood cells, greatly improves the transport capacity and efficiency of the nanocarrier system in the body, and thus improves the bioavailability of dihydroartemisinin in the body and enhances the antimalarial effect. At the same time, the physical characterization of heparin-dihydroartemisinin liposome nanoparticles is good, the toxicity to cells is low, the biocompatibility is good, and it meets the safety indicators for intravenous injection.

[0015] The present application provides a new effective means for malaria treatment, and also provides a new idea and platform for the research and development of antimalarial drugs. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 Figure (a) is a transmission electron microscope measurement result of the present application, and figure (b) is a detection result of the present application by dynamic light scattering (DLS) and photoluminescence spectrum (PLAS); Figure 2 Figure is a stability test result of HEP-DHA@Lips for one month in the present application; Figure 3 Figure is a typical drug sustained release curve of HEP-DHA@Lips nanoparticles obtained in the drug release test of the present application; Figure 4 Figure is a result analysis chart and a real photo obtained in the blood compatibility test of HEP-DHA@Lips nanoparticles in the present application; Figure 5 Figure is a result chart obtained in the cytotoxicity test of HEP-DHA@Lips nanoparticles in the present application; Figure 6 Figure is a fluorescence localization chart of HEP-Dil@Lips binding to malignant malaria schizonts and schizonts in the present application; Figure 7 Figure (a) is an IC50 curve of HEP-DHA@Lip, DHA@Lip without heparin coating and DHA raw drug for inhibiting in vitro cultured malignant malaria, and figure (b) is a significance analysis column chart in the present application; Figure 8 Figure (a) is a mouse infection rate line chart of HEP-DHA@Lip in the mouse malaria treatment evaluation in the present application, and figure (b) is a survival curve chart. DETAILED DESCRIPTION

[0017] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with the accompanying drawings and embodiments.

[0018] like Figures 1-8 As shown, the preparation method of the heparin-targeted dihydroartemisinin-loaded antimalarial nanoliposome lyophilized powder drug provided in this application specifically includes the following steps: S1: Synthesis of DSPE-PEG2000-Heparin: Dalteparin was coupled with DSPE-PEG2000-NHS via an amination reaction, purified by dialysis, and lyophilized. S2: Dissolve soybean lecithin, cholesterol, DSPE-PEG2000-heparin polymer and dihydroartemisinin in 5 mL of chloroform, evaporate to form a film under low pressure using a rotary evaporator, and then add deionized water to dissolve. S3: Deionized water was added for membrane hydration, followed by sonication in a 20°C water bath for 1 min and extrusion through a 100nm membrane to obtain HEP-DHA@Lips. The membrane was then dialyzed in deionized water for 4 h to remove unloaded DSPE-PEG2K-heparin and dihydroartemisinin.

[0019] S4: Add a freeze-drying protectant at a mass ratio of 1:5, freeze-dry, and obtain HEP-DHA@Lips freeze-dried powder formulation.

[0020] Before step S1, material preparation and experimental animal preparation are required. Material preparation includes preparing dihydroartemisinin (DHA, purity ≥99%), DSPE-PEG2000-NHS (1,2-distearate-sn-glycerol-3-phosphate ethanolamine-n-2000), low molecular weight heparin (MW~6000), powdered RPMI-1640, hypoxanthine monohydrochloride, gentamicin sulfate (cell culture grade), DMEM, penicillin-streptomycin, AlbuMAXII, and fetal bovine serum, etc. The experimental water is Milli-Q grade water that has been sterilized by ultraviolet light. Female BALB / c mice (weighing 18-20 grams) aged five to six weeks were selected for the experimental animals and fed standard feed on an ad libitum basis. One week before the experiment, the feeding procedures for all experimental animals were carried out in accordance with the animal feeding guidelines of Shenyang Agricultural University.

[0021] The lyophilized powder preparation includes soy lecithin, cholesterol, DSPE-PEG2000-heparin polymer and dihydroartemisinin, wherein the molar ratio of each component is 10:2:1:1, and the lyophilized powder is prepared after the lyophilized powder is treated with a lyophilized protective agent. And the particle size range is 125.42±3.24nm, and the absolute value of Zeta potential is more than 20mV, and it has good hydrophilicity. In addition, the nano drug exists in the form of lyophilized powder, and after being stored at 4℃ for one month, it still maintains stable particle size, potential and drug loading capacity, indicating that it has good preparation stability and potential for further conversion and application.

[0022] In summary, the anti-malaria nano-liposome lyophilized powder drug is composed of dihydroartemisinin with a loading capacity of 10.58±0.67%, liposome and DSPE-PEG2000-heparin liposome nanocarrier system, which is prepared by conventional thin film hydration method, and heparin is coated on the outer layer of the nanoparticles to form HEP-DHA@Lips structure. Among them, the inside is a liposome structure, which loads dihydroartemisinin; the outside is coated with DSPE-PEG2000-heparin, which can realize the specific recognition and enrichment of infected malaria red blood cells by taking advantage of the specific recognition of heparin to malaria schizonts or infected red blood cells. EMBODIMENT

[0023] The application provides a use of a heparin-targeted dihydroartemisinin-loaded anti-malaria nano-liposome lyophilized powder drug for malaria treatment, which can inhibit the proliferation of malaria parasites in blood and prolong the survival time of infected hosts, improve the bioavailability of dihydroartemisinin in vivo, and enhance the anti-malaria effect. EMBODIMENT

[0024] As shown in Figures 1-3 , high-resolution morphological analysis of HEP-DHA@Lips prepared in Example 1 was performed using a transmission electron microscope (TEM, HR-TEM7800, Hitachi, Tokyo, Japan) in bright-field imaging mode; the synthesized HEP-DHA@Lip nanoparticles were detected based on dynamic light scattering (DLS) and phase analysis light scattering (PALS).

[0025] As shown in Figure 1 , the HEP-DHA@Lips prepared in Example 1 is spherical and has good dispersibility, as shown in Figure 1 , the average particle size is about 125.42±3.24nm, and the absolute value of Zeta potential is more than 20, indicating that it has good hydrophilicity; EMBODIMENT

[0026] The drug loading efficiency and in vitro drug release test of HEP-DHA@Lips nanoparticles were determined by high performance liquid chromatography (HPLC): The HEP-DHA@Lips lyophilized powder was dispersed in ultrapure water (1 mg / mL), and the drug loading efficiency was quantified by HPLC analysis, with the calculation formula being: Drug loading efficiency (%) = detected mass of dihydroartemisinin / total mass of nanoliposomes x 100% Release test: HEP-DHA@Lips (1 mg / mL, prepared in ultrapure water) was encapsulated in a dialysis bag (MW 8-14 kDa) and immersed in a release medium containing 0.1% Tween-80 (pH 5.6 or 7.4, 37°C, 100 rpm) for 48 hours. At predetermined time points (0, 2, 4, 8, 12, 24, and 48 hours), 1 mL of buffer was sampled, and an equal amount of fresh medium was supplemented. The drug release efficiency calculation formula is: Release efficiency (%) = detected mass of DHA in the release medium / theoretical mass of DHA in the nanoliposomes x 100% Table 1 shows the drug loading rate detection results of HEP-DHA@Lips

[0027] As shown in Table 1, the average drug loading rate (%) of heparin-dihydroartemisinin liposomes is 10.58 ± 0.67; Determination of DHA content in HEP-DHA@Lips by high performance liquid chromatography: the ultraviolet detection wavelength is 234 nm. A C18 chromatographic column (5 μm, 4.6 x 250 mm) is used, the mobile phase is acetonitrile and (30:70, v / v, pH 2.5) at a flow rate of 1.0 mL / min. Specifically, all collected samples were mixed with anhydrous ethanol and 0.2% NaOH solution (0.2:1:4, v / v), then incubated in the dark at 56°C for 10 minutes. Standard curves were prepared using DHA ethanol solutions of different concentrations (5, 10, 15, 20, 30 μg / mL) by the same process. Finally, the samples were mixed with the mobile phase at a volume ratio of 1:4 before measurement, and the loading amount was 20 μL.

[0028] As shown in Figure 3 , the in vitro release experiment shows that in a buffer at pH 5.6 (simulating the acidic environment of malaria-infected cells), dihydroartemisinin in HEP-DHA@Lips can be released slowly over time within 48 hours, with a release efficiency higher than that in a pH 7.4 environment (≥80%). The above results not only reflect the pH-responsive release characteristics, but also significantly extend the half-life of dihydroartemisinin by about 5 times, greatly improving the time of dihydroartemisinin and malaria in the body.

[0029] Therefore, HEP-DHA@Lips possesses extremely high transproduction potential. In vitro release experiments showed that in a pH 5.6 buffer solution (simulating the acidic environment of Plasmodium-infected cells), dihydroartemisinin from HEP-DHA@Lips could be slowly released over 48 hours, with a release efficiency higher than that in the pH 7.4 environment (≥80%). These results not only demonstrate pH-responsive release characteristics but also significantly extend the half-life of dihydroartemisinin by approximately 5 times, greatly enhancing the duration of the antiparasitic effect of dihydroartemisinin and Plasmodium in vivo. Example

[0030] The test was conducted at 4°C for one month. Specifically, the zeta potential, nanoparticle size, and dispersion concentration of HEP-DHA@Lips were analyzed using dynamic light scattering particle size analyzer and high performance liquid chromatography on days 1, 3, 5, 7, 15, and 30. Three parallel samples were tested at each time point after being dissolved in deionized water at a ratio of 1:20.

[0031] like Figure 2 As shown, where a represents the change in drug loading rate of HEP-DHA@Lips, b represents the change in size of HEP-DHA@Lips, and c represents the change in potential of HEP-DHA@Lips, after storage at 4°C for one month, the drug loading rate, particle size, and encapsulation efficiency of HEP-DHA@Lips showed no significant changes, indicating excellent stability under normal storage conditions. This further demonstrates that the transportation or storage requirements of HEP-DHA@Lips in this application can be met in malaria-prone areas, especially in some developing countries or impoverished regions. Therefore, HEP-DHA@Lips has extremely high potential for conversion to other production processes. Example

[0032] like Figure 4 and Figure 5 As shown, the cytotoxicity of heparin-dihydroartemisinin liposome nanoparticles against HepG2 human hepatocellular carcinoma cells was evaluated. The hemolytic activity of the heparin-dihydroartemisinin liposome nanoparticles was assessed by co-incubating different concentrations (15-500 μg / mL) of HEP-DHA@Lips solution with 4% hematocrit human erythrocyte suspension. Details are as follows: Fresh DMEM medium (containing 8% fetal bovine serum) was used to prepare HEP-DHA@Lips solutions with different concentrations (15-500 μg / mL), which were co-incubated with HEPG2 cells for 48 hours. Then, a CCK-8 kit was used to detect the possible cytotoxicity of HEP-DHA@Lips nanoparticles. The intravenous injection safety of HEP-DHA@Lips nanoparticles was evaluated by a human red blood cell hemolysis test. Different concentrations (15-500 μg / mL) of HEP-DHA@Lips solutions were used to incubate 4% hematocrit normal human red blood cell suspensions (prepared using complete medium) at 37°C for 2 hours. After centrifugation at 1500 rpm for 5 minutes to precipitate the red blood cells, the supernatant was taken and the absorbance was measured at 400 nm using a microplate reader.

[0033] As shown in Figure 4 , the hemolysis rate of HEP-DHA@Lips was ≤1.5%, which met the intravenous injection safety standard of biomedical materials (hemolysis rate ≤5%). This indicated that the HEP-DHA@Lips synthesized in the application could be used for the treatment of blood stage of Plasmodium by intravenous injection, directly targeting and killing the parasites in the blood. Compared with the traditional oral administration, the intravenous injection of HEP-DHA@Lips preparation not only avoids the problem of large amount of administration but little effect in clinical practice, but also overcomes the problems of short half-life of artemisinin drugs, long treatment course, malaria relapse, etc., achieving efficient treatment of malaria.

[0034] The cytotoxicity detection results showed that after co-culturing with HEP-DHA@Lips for 48 hours, the survival rate of HepG2 cells remained above 80% even at a concentration as high as 500 μg / mL. In addition to excluding the possible influence of drug-loaded nanoparticles on cell growth and development, the cytotoxicity of HEP-DHA@Lips was almost negligible Embodiment

[0035] As shown in Figure 6 , the interaction between HEP-Dil@Lips and Plasmodium falciparum merozoites and infected red blood cells was analyzed by fluorescence localization, and the specific content was as follows: HEP-DHA@Lips was replaced by red fluorescent dye Dil to successfully prepare HEP-Dil@Lips nanoparticles with red fluorescent labeling. After incubation with P. falciparum at different developmental stages for 4 hours at 37°C in the dark, centrifugation at 3000 rpm for 5 minutes, and three times of co-washing with equal volume of RPMI-1640 complete medium, the malaria parasite nuclei were stained with blue fluorescent dye DAPI; finally, the enrichment of red fluorescence in HEP-Dil@Lips nanoparticles on the schizonts and infected red blood cells was observed using a structured illumination combined with super-resolution microscope (SIM-ultimate, Guangzhou Zhongke Rui Biological Technology Co., Ltd.).

[0036] The fluorescence co-localization experiment with P. falciparum showed that HEP-Dil@Lips had obvious red fluorescence enrichment on the surface of P. falciparum schizonts and schizonts, further proving the targeting effect of the heparin-modified liposome nanoparticles synthesized in the application on P. falciparum and its infected red blood cells. EMBODIMENT

[0037] Different concentrations of DHA@Lips and HEP-DHA@Lips (40 ng / mL to 0.08 ng / mL, 2-fold dilution ratio) were prepared using complete medium; the raw drug of dihydroartemisinin was dissolved in anhydrous ethanol to prepare a 1 mg / mL stock solution, which was then diluted in complete medium to prepare working solutions of different concentrations.

[0038] The above solutions were sequentially added to the culture containing highly synchronized P. falciparum (infected rate 0.5%) at a volume ratio of 1:1, and incubated at 37°C for 48 hours; P. falciparum cultured in normal complete medium was used as a negative control; after DHE staining, the infected rate was detected using a BD FACSAria III flow cytometer; the data were analyzed and calculated using the sigma dose-response equation in GraphPad Prism 9 software.

[0039] As shown in Figure 7 , the dose-response regression curve and value comparison analysis in the in vitro insecticide activity evaluation can confirm that, compared with natural DHA ( = 0.98 ± 0.20 nM), HEP-DHA@Lips ( = 0.32 ± 0.044 nM) and non-heparin-targeted DHA@Lips ( ​Both (0.52±0.042 nM) showed stronger antimalarial activity. The superior efficacy of HEP-DHA@Lips highlights the role of heparin as an effective targeting element, which helps to specifically bind to infected red blood cells, thereby promoting more effective drug delivery and release. HEP-DHA@Lips not only inherits the antimalarial activity of dihydroartemisinin, but also significantly improves its parasite-inhibiting efficacy by about four times, optimizing the parasite-inhibiting concentration of dihydroartemisinin to the picomolar concentration level, achieving highly efficient killing of malaria parasites. Example

[0040] The antimalarial activity of HEP-DHA@Lips in a mouse malaria model was evaluated, and the details are as follows: Using the "Pitt four-day inhibition test", BALB / c mice were randomly divided into three groups (n=8 per group), and each mouse was injected intraperitoneally with 1× Red blood cells infected with *P. berghei* ANKA were collected. Treatment began 72 hours post-infection (parasitemia 0.5% to 1%), with mice receiving dihydroartemisinin at a dose of 2 mg / mouse / day for four consecutive times. Three groups of mice received HEP-DHA@Lips via tail vein injection, dihydroartemisinin via gavage, and saline via tail vein injection, respectively. Dihydroartemisinin solution was prepared using carboxymethyl cellulose (CMC) solvent and stirred at 4°C in the dark for more than 24 hours to ensure complete dissolution. Parasitemia in infected mice was monitored every other day by Giemsa-stained blood smears, and mouse survival was recorded daily.

[0041] like Figure 8 As shown, in a mouse malaria treatment model, the synthesized HEP-DHA@NEs, after four consecutive days of tail vein injection, exhibited excellent in vivo antimalarial effects. Compared with the saline group and the control group with an equal amount of dihydroartemisinin, mice in the HEP-DHA@NEs treatment group showed a significant reduction in parasitemia during the treatment period. During the treatment period, mice in the HEP-DHA@NEs treatment group achieved a 100% parasitemia-negative conversion rate, and the survival time of all mice in this group was significantly prolonged, with a survival rate of 87.5% during the observation period (≥60 days). These results fully demonstrate that HEP-DHA@NEs possesses significant therapeutic advantages and conversion potential in malaria treatment.

[0042] All results are expressed as mean ± standard deviation (n≥3). Statistical significance analysis was performed using the Student's t-test, and graph generation and statistical calculations were completed using GraphPadPrism9 software.

[0043] The testing methods described in this application may vary depending on the animal species, research objectives, and experimental conditions. Therefore, appropriate adjustments and optimizations should be made according to actual needs in specific research.

[0044] The application also provides a pharmaceutical composition for treating malaria, comprising the above-mentioned nano-drug and pharmaceutically acceptable adjuvant or solvent. The pharmaceutical composition adopts an intravenous injection form, the adjuvant comprises physiological saline for injection, CMC, etc., and the pH of the pharmaceutical composition is adjusted to 6.8-7.4.

[0045] It is apparent to those skilled in the art that the application is not limited to the details of the foregoing exemplary embodiments, and that the application can be implemented in other particular forms without departing from the spirit or essential characteristics of the application. Therefore, the embodiments should be considered in all respects as illustrative and not restrictive, and the scope of the application should be defined by the appended claims rather than the above description, and it is intended to include all changes falling within the meaning and scope of the equivalent elements of the claims. Any reference signs in the claims should not be considered as limiting the claims involved.

[0046] Furthermore, it should be understood that although the present specification is described in terms of embodiments, not every embodiment contains only one independent technical solution, and the description of the specification is only for the sake of clarity, and those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can be appropriately combined to form other embodiments that those skilled in the art can understand.

Claims

1. A lyophilized powder of antimalarial nano-liposome loaded with dihydroartemisinin targeting heparin, characterized in that, The liposome nano-drug delivery system is composed of dihydroartemisinin, liposome and DSPE-PEG2000-heparin, and is prepared by a conventional thin film hydration method, and the heparin is coated on the outer layer of the nanoparticles, and a freeze-drying protective agent is added to obtain a HEP-DHA@Lips freeze-dried powder preparation.

2. The heparin-targeted dihydroartemisinin-loaded antimalarial nanoliposome lyophilized powder drug according to claim 1, characterized in that, The particle size of the HEP-DHA@Lips freeze-dried powder preparation is 125.42±3.24nm, and the absolute value of Zeta potential is more than 20mV.

3. The heparin-targeted dihydroartemisinin-loaded antimalarial nanoliposome lyophilized powder drug according to claim 1, characterized in that, The dihydroartemisinin drug loading capacity is 10.58±0.67%.

4. A process for the preparation of a lyophilized powder of antimalarial nano-liposomes of dihydroartemisinin targeted to heparin, characterized in that, The method comprises the following steps: S1: synthesis of DSPE-PEG2000-heparin: coupling of heparin and DSPE-PEG2000-NHS by amination reaction, dialysis purification and freeze-drying; S2: dissolving soybean lecithin, cholesterol, DSPE-PEG2000-heparin polymer and dihydroartemisinin in 5mL chloroform, evaporating into a film under low pressure, and then adding deionized water for dissolution; S3: adding deionized water for film hydration, and ultrasonic treatment in a 20℃ water bath for 1min, and extruding through a 100nm membrane to obtain HEP-DHA@Lips; S4: adding a freeze-drying protective agent, and freeze-drying to obtain a HEP-DHA@Lips freeze-dried powder preparation.

5. The process for the preparation of a lyophilized powder of antimalarial nano-liposomes of dihydroartemisinin targeted to heparin according to claim 4, characterized in that, After obtaining the HEP-DHA@Lips in S3, it needs to be dialyzed in deionized water for 4h to remove the DSPE-PEG2K-heparin and dihydroartemisinin that are not successfully loaded.

6. The process for the preparation of a lyophilized powder of antimalarial nano-liposomes of dihydroartemisinin targeted to heparin according to claim 4, characterized in that, The mass ratio of HEP-DHA@Lips to freeze-drying protective agent in S4 is 1:

5.

7. A process for the preparation of a lyophilized powder of a liver-targeted bilastatine-loaded antimalarial nanoliposome drug according to claim 4, characterized by the fact that, The preparation comprises soybean lecithin, cholesterol, DSPE-PEG2000-heparin polymer and dihydroartemisinin, and the molar ratio of each component is 10:2:1:

1.

8. Use of a lyophilized powder of a heparin-targeted dihydroartemisinin-loaded antimalarial nanoliposome drug, characterized in that, It is used for inhibiting the proliferation of malaria parasites in blood and prolonging the survival time of infected hosts.

Images