Preparation method and application of drug delivery carrier based on dunaliella salina cells

Abstract

The invention relates to the technical field of biological medicine, in particular to a preparation method and application of a drug delivery carrier based on dunaliella salina cells, and the method comprises the following steps: selecting dunaliella salina for standardized culture to obtain dunaliella salina living cells; the target drug and the dunaliella salina living cells are loaded into the dunaliella salina living cells through an endocytosis method; free drug molecules are removed through centrifugation, and the dunaliella salina cell drug delivery carrier is obtained. Efficient drug loading is achieved by activating an endocytosis pathway, the highest quercetin loading rate reaches 92%, and the quercetin loading rate is remarkably superior to that of a co-incubation method and an electroporation method; the dunaliella salina cell carrier is excellent in biocompatibility, and the cell survival rate is gt; no organ toxicity exists in a mouse experiment; the medicine can be slowly released in intestinal tracts, the retention time is prolonged, the anti-tumor effect in a colorectal cancer model is remarkable, and the immunoregulation effect can be enhanced; the preparation process is simple and controllable, special equipment is not needed, and a new scheme is provided for constructing an efficient delivery system for natural drugs.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a preparation method and application of a drug delivery carrier based on Dunaliella cells. Background Art

[0002] With the rapid development of biomedical technology, nanoscale drug delivery systems have become an important means to improve drug efficacy and reduce side effects. In the field of cancer treatment, nanoparticle drug carriers have attracted much attention due to their unique targeting and high efficiency. However, existing drug carriers, such as synthetic carriers based on organic and inorganic materials, while improving drug delivery efficiency to a certain extent, still have many problems during use.

[0003] Specifically, synthetic carriers often have drawbacks such as high toxicity, susceptibility to immune responses, and rapid clearance from the body, which limit their widespread clinical application. In contrast, exosomes, as natural small membrane vesicles, have nanoscale dimensions, good biocompatibility, and the ability to specifically bind to receptor cells, and are therefore considered ideal drug carriers.

[0004] Exosomes are small membrane vesicles secreted by various cells under normal and pathological conditions, containing a variety of RNA, proteins, and lipid molecules. Among them, disc-shaped vesicles with a diameter of 40-100 nm have attracted particular attention. Exosomes not only serve as a medium for intercellular communication but also as a drug delivery system, precisely delivering drugs to target cells for effective treatment.

[0005] However, exosomes from different cell sources vary significantly in composition, stability, homogeneity, and controllability of their multi-component structure. This presents numerous challenges in their preparation and application. For example, the low yield of exosomes from certain cell sources makes it difficult to meet the demands of large-scale clinical trials. Furthermore, the complex composition of exosomes makes it difficult to ensure their stability and targeting in vivo.

[0006] To overcome these shortcomings, Chinese invention patent application number 201910816594.6 proposes a method for preparing a targeted drug delivery vector based on Dunaliella exosomes. This method achieves precise targeted drug delivery by culturing Dunaliella, extracting Dunaliella exosomes, chemically coupling the targeting peptide to the Dunaliella exosomes, and loading the drug onto the Dunaliella exosomes via electroporation. However, this method still has some shortcomings. For example, when using electroporation to load drugs, the high voltage can damage the structural integrity of biomacromolecules, resulting in loss of drug activity and limited drug loading efficiency. In the targeted delivery process, the targeting peptide is chemically coupled to the vector. However, the targeting peptide is highly susceptible to enzymatic degradation in the complex physiological environment of the body. Various enzymes in the body recognize and cleave specific chemical bonds of the targeting peptide, causing it to fall off the vector, significantly reducing the targeting efficiency of the vector. Summary of the Invention

[0007] The present invention aims to overcome the shortcomings of the prior art by providing a method for preparing a drug delivery vehicle based on Dunaliella cells and its application. This method achieves efficient drug loading through endocytosis, leveraging the natural biocompatibility and intestinal adsorption properties of Dunaliella cells to construct a drug delivery system with high loading rate, low toxicity, and long-term sustained release, addressing the low drug delivery efficiency and poor targeting characteristics of the prior art.

[0008] The present invention provides a method for preparing a drug delivery carrier based on Dunaliella cells, comprising the following steps:

[0009] S1: Dunaliella salina was selected for standardized culture to obtain living cells of Dunaliella salina;

[0010] S2: The target drug and the living cells of Dunaliella are loaded into the living cells of Dunaliella through endocytosis;

[0011] S3: Centrifuge to remove free drug molecules and obtain Dunaliella cell drug delivery carrier.

[0012] Preferably, the step S1 specifically includes:

[0013] S101: preparing modified Dunaliella culture medium;

[0014] S102: In a conical flask, at 26°C and a light intensity of 50 μM photons·m -2 ·s -1 , culturing Dunaliella salina cells in a modified Dunaliella medium under a light intensity of 12 hours per day;

[0015] S103: When the cells grew to the logarithmic phase, they were collected by centrifugation;

[0016] S104: Wash the cells three times with fresh medium and adjust the cell concentration to 5 × 10 6 / mL;

[0017] S105: Centrifuge the treated cell mixture at a rotation speed of 2500 rpm for 5 minutes to obtain live Dunaliella cells.

[0018] Preferably, the modified Dunaliella culture medium in step S101 comprises:

[0019] 1.5M NaCl, 1μM CuCl·2H2O, 10mM KNO3, 50mM NaHCO3, 0.4mM KHPO4, 185μM H3BO3, 2μM FeCl6·H2O, 5mM MgSO4·7H2O, 5μM EDTA, 1μM (NH4)MO7O 24 ·4H2O, 7μM MnCl2·4H2O, 1μM ZnCl2, 1μM CoCl26·6H2O and 0.2mM CaCl2.

[0020] Preferably, in the logarithmic stage in step S103, the concentration of Dunaliella cells is 1×10 6 cells / mL to 1×10 7 pieces / mL.

[0021] Preferably, the step S2 specifically includes:

[0022] S201: Pretreatment of Dunaliella live cells

[0023] a. Prepare a pretreatment solution containing 2.5 mM ATP, 2.5 mM MgCl2, 1 mM CaCl2;

[0024] b. Incubate 1 volume of Dunaliella live cells with 9 volumes of the pretreatment solution at room temperature for 5 minutes;

[0025] c. Centrifuge at 2500 rpm for 5 minutes to obtain the pretreated Dunaliella cells;

[0026] S202: Target drug loading

[0027] a. Dissolve the target drug in 154 mM NaCl solution;

[0028] b. Incubate the Dunaliella cells obtained in step S201 with the target drug solution at 37°C for 10 minutes;

[0029] c. Centrifuge at 2500 rpm for 5 minutes to obtain the Dunaliella cell drug delivery vector.

[0030] Preferably, the target drug is quercetin, curcumin or liquiritin.

[0031] The present invention also provides a salt algae cell drug delivery carrier prepared by using the preparation method of the salt algae cell-based drug delivery carrier.

[0032] The present invention also provides an application of the salt algae cell drug delivery carrier in preparing a D. salina-QCT delivery carrier.

[0033] The present invention also provides an application of the salt algae cell drug delivery carrier in preparing a D. salina-curcumin delivery carrier.

[0034] The present invention also provides an application of the salt algae cell drug delivery carrier in preparing a D. salina-liquiritin delivery carrier.

[0035] Beneficial effects

[0036] The salt algae cell drug delivery carrier prepared by the cell endocytosis method of the present invention not only has a high loading efficiency, with a quercetin loading rate of up to 92%, which is significantly better than the 60% of the co-incubation method and the 40% of the electroporation method, but also has universal applicability to a variety of drugs such as curcumin and liquiritin; it also has excellent biocompatibility, with a cell survival rate of over 90%, and no pathological damage to organs and no significant changes in body weight in mouse experiments; at the same time, the salt algae cells can be adsorbed on the intestinal villi to achieve long-term sustained release of drugs and prolong the residence time of drugs in the intestine. In a mouse in situ colorectal cancer model, the anti-tumor effect of the carrier after loading quercetin is significantly better than that of the free drug, and it can further inhibit the proportion and function of MDSC cells in the tumor and enhance the immune regulation effect; in addition, the preparation method is simple and controllable, does not require complex equipment, the culture medium composition and culture conditions are clear, the cost is low, and the repeatability is strong, which effectively overcomes the defects of existing synthetic carriers such as high toxicity, poor stability of exosome preparation and low loading efficiency of electric shock method. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is a diagram showing the drug loading efficiency of quercetin at different concentrations when D.salina-QCT is prepared by the endocytosis method of the present invention;

[0038] Figure 2 This is a diagram showing the drug loading efficiency of quercetin at different concentrations when D.salina-QCT is prepared using the co-incubation method of the present invention;

[0039] Figure 3 This is a diagram showing the drug loading efficiency of quercetin at different concentrations when D.salina-QCT was prepared by the electroporation method of the present invention;

[0040] Figure 4These are in vivo imaging images of drug distribution in mice at different time points after oral administration of QCT-FITC and D. salina-QCT-FITC according to the present invention;

[0041] Figure 5 This is a statistical line graph of the fluorescence intensity of the drug at different time points after oral gavage of the present invention;

[0042] Figure 6 This is an immunofluorescence staining image of the co-localization of Dunaliella cells and QCT-FITC in the intestinal villi;

[0043] Figure 7 This is an electron microscope observation of the combination of Dunaliella cells and drugs in the intestinal villi;

[0044] Figure 8 The figures are live imaging images of tumor growth in each group in the mouse orthotopic colorectal cancer model of the present invention;

[0045] Figure 9 Statistical graph of tumor volume in each group in the mouse orthotopic colorectal cancer model of the present invention;

[0046] Figure 10 This is a curve chart of dynamic monitoring of mouse body weight during the drug administration period of the present invention;

[0047] Figure 11 HE staining of the heart, liver, spleen, lung and kidney of the mouse of the present invention;

[0048] Figure 12 This is a diagram showing the proportion and function detection results of MDSC cells in tumors according to the present invention;

[0049] Figure 13 This is a graph showing the loading efficiency of curcumin at different concentrations into Dunaliella cells via the endocytosis method of the present invention;

[0050] Figure 14 This is a diagram showing the efficiency of loading different concentrations of liquiritin into Dunaliella cells via endocytosis. DETAILED DESCRIPTION

[0051] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0052] All other embodiments obtained by persons of ordinary skill in the art based on the embodiments herein without inventive effort are intended to fall within the scope of protection of the present invention. Experimental methods in the following examples, where specific conditions are not specified, were generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise indicated, all percentages, ratios, proportions, and parts are by weight.

[0053] Unless otherwise specified, the reagents and raw materials used in the examples and comparative examples of the present invention can be obtained through commercial channels.

[0054] Example 1: Preparation of D. salina-QCT by endocytosis

[0055] (1) Preparation of modified Dunaliella culture medium

[0056] Weigh 1.5M NaCl, 1μM CuCl·2H2O, 10mM KNO3, 50mM NaHCO3, 0.4mM KHPO4, 185μMH3BO3, 2μM FeCl6·H2O, 5mM MgSO4·7H2O, 5μM EDTA, 1μM (NH4)MO7O 24 ·4H2O, 7μM MnCl2·4H2O, 1μM ZnCl2, 1μM CoCl26·6H2O and 0.2mM CaCl2 were dissolved in deionized water and the volume was adjusted to 2L.

[0057] (2) Dunaliella salina cell culture

[0058] Dunaliella salina cells were inoculated into a 2 L conical flask, and the above culture medium was added. The culture medium was incubated at 26 °C and a light intensity of 50 μM photons·m -2 ·s -1 and a light intensity of 12 h / day until the cells grew to the logarithmic phase.

[0059] (3) Cell collection and washing

[0060] The cultured cell suspension was centrifuged at 2500 rpm for 5 minutes, the supernatant was discarded, and the cells were washed three times with fresh culture medium, each time under the same centrifugation conditions; after washing, the cells were resuspended in culture medium and the concentration was adjusted to 5×10 6 pieces / mL.

[0061] (4) Pretreatment of Dunaliella cells

[0062] Prepare pretreatment solution: an aqueous solution containing 2.5 mM ATP, 2.5 mM MgCl2, and 1 mM CaCl2;

[0063] Take 1mL of the solution with a concentration of 5×10 6 9 mL of pretreatment solution was added to a suspension of Dunaliella cells at 1000 nm / mL and incubated at room temperature for 5 minutes; the suspension was then centrifuged at 2500 rpm for 5 minutes, the supernatant was discarded, and the pretreated Dunaliella cells were collected.

[0064] (5) Quercetin (QCT) loading and complex preparation

[0065] Quercetin was dissolved in 154 mM NaCl solution to prepare a drug solution with a concentration of 6.104 μM;

[0066] The pretreated D. salina cells were resuspended in a quercetin solution and incubated at 37° C. for 10 minutes. After the incubation, the cells were centrifuged at 2500 rpm for 5 minutes, and the supernatant was discarded to obtain quercetin-loaded D. salina-QCT.

[0067] Example 2

[0068] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 12.207 μM.

[0069] Example 3

[0070] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 24.414 μM.

[0071] Example 4

[0072] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 48.828 μM.

[0073] Example 5

[0074] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 97.656 μM.

[0075] Example 6

[0076] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 195.313 μM.

[0077] Example 7

[0078] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 390.625 μM.

[0079] Example 8

[0080] The difference from Example 1 is that different concentrations of quercetin drug solutions are used: the concentration of the drug solution is 781.25 μM.

[0081] Example 1-8 investigated the effects of different concentrations of quercetin drug solutions on the drug loading rate of Dunaliella cells by endocytosis. Figure 1As shown, when the quercetin drug concentration gradually increased to 390.625μM, the drug loading rate of Dunaliella cells reached a peak of approximately 92%. However, when the concentration continued to increase to 781.25μM, the loading rate did not show a significant increase and may even stabilize or slightly decrease. This indicates that there is an optimal drug concentration adaptation range for the cellular endocytosis method. 390.625μM quercetin can achieve efficient loading by activating the endocytosis mechanism of Dunaliella cells. This provides an experimental basis for concentration optimization for the preparation of Dunaliella cell drug delivery vectors and verifies the loading efficiency advantage of this method at specific drug concentrations.

[0082] Comparative Example 1: Preparation of D.salina-QCT by co-incubation method

[0083] Dunaliella cells were collected from the cultured Dunaliella salina, the culture medium was removed by centrifugation (2500 rpm, 5 min), and the cell concentration was adjusted to 5 × 10 6 / mL;

[0084] Quercetin solutions of different concentrations were prepared: 48.82 μM, 97.6 μM, 195.31 μM, 390.62 μM, and 781.25 μM, respectively, and dissolved in 154 mM NaCl isotonic solution;

[0085] Take 1 mL of Dunaliella cell suspension and add 9 mL of the above quercetin solution of different concentrations;

[0086] Incubate at 37°C in the dark for 24 hours;

[0087] After the incubation, the mixture was centrifuged at 2500 rpm for 5 minutes, and the supernatant was discarded. Free QCT drug molecules that were not loaded into the D. salina cells were removed by centrifugation to obtain D. salina-QCT loaded with quercetin.

[0088] Comparative Example 2: Preparation of D.salina-QCT by electroporation

[0089] Dunaliella cells were collected from the cultured Dunaliella salina, the culture medium was removed by centrifugation (2500 rpm, 5 min), and the cell concentration was adjusted to 5 × 10 6 / mL;

[0090] A 390.62 μM quercetin solution was prepared by dissolving it in an isotonic 154 mM NaCl solution;

[0091] Mix 100 μL of Dunaliella cell suspension with 100 μL of quercetin solution and add to the electroporation cup;

[0092] Electroporation was performed under different voltage conditions (100 V, 200 V, 300 V, 400 V, 500 V, 600 V, and 700 V), and the parameters were set as follows: single pulse, pulse width 20 ms;

[0093] After electroporation, the cell suspension was transferred to isotonic solution at 37°C and incubated for 1 h;

[0094] After the incubation, the mixture was centrifuged at 2500 rpm for 5 minutes, and the supernatant was discarded. Free QCT drug molecules that were not loaded into the Dunaliella cells were removed by centrifugation to obtain D. salina-QCT loaded with quercetin.

[0095] In the above embodiment, the co-incubation method was used to prepare D.salina-QCT, and the drug loading rate thereof could reach up to about 60%. Figure 2 As shown. The drug loading was performed by electroporation. The drug loading rate was the highest at a voltage of 600V and a capacitance of 50μF, about 40%. Figure 3 When the quercetin concentration was 390.625 μM, the drug loading rate using the endocytosis method was about 92%, as shown in Figure 1 shown.

[0096] It can be seen that in terms of quercetin loading efficiency, the endocytosis method of the present application is significantly superior to the co-incubation method and the electroporation method. From the perspective of the mechanism of action, the endocytosis method activates the natural endocytosis pathway of Dunaliella cells through the culture medium to achieve efficient drug embedding; while the co-incubation method relies on the passive diffusion of the drug, and the electroporation method may damage cell activity due to the voltage, thereby limiting the loading efficiency. Experimental data show that the endocytosis method shows obvious advantages in loading efficiency by optimizing drug concentration and endocytosis induction conditions on the basis of maintaining cell activity, providing a more efficient and low-loss feasible solution for the preparation of Dunaliella cell drug delivery carriers.

[0097] In vivo metabolic detection after drug loading in Dunaliella cells:

[0098] First, QCT was labeled with FITC to detect the metabolism of the drug in the body under loaded and unloaded conditions. QCT-FITC and D. salina-QCT-FITC were used for gavage respectively, and the distribution of the drug was detected by small animal in vivo imaging at 0h, 1.5h, 3h, 4.5h, and 6h after gavage. Figure 4 The fluorescence is statistically analyzed to reflect the concentration of the drug, as shown in Figure 5 As shown. The results showed that after loading the living cells of Dunaliella with drugs, not only did the drugs stay in the intestine for a longer period of time, but they also achieved the effect of sustained drug release. Immunofluorescence and electron microscopy results showed that Dunaliella and drugs were co-localized on the intestinal villi, as shown in Figure 2. Figure 6-7 The results show that Dunaliella can be adsorbed on the intestinal villi and slowly broken down, thus promoting the absorption of drugs.

[0099] To test the in vivo anti-colorectal cancer effect of live D. salina cells loaded with quercetin, an orthotopic colorectal cancer model was established in mice. The mice were divided into four groups: Con, D. salina, QCT, and D. salina-QCT, and were administered with drugs. Tumor growth was monitored using in vivo imaging of small animals. The results showed that the D. salina-loaded quercetin group had the best anti-colorectal cancer growth effect. Figure 8-9 The body weight of mice was monitored dynamically during the administration period. Figure 10 As shown, at the end of the experiment, the mice were taken for HE staining of heart, liver, spleen, lung and kidney. Figure 11 The results showed that the drug and Dunaliella had no toxic side effects on mice. Detection of MDSC cells inside the tumor revealed that quercetin could effectively reduce the proportion and function of MDSC cells inside the tumor. However, after Dunaliella was loaded with quercetin, it could further inhibit the proportion and function of MDSC cells inside the tumor. Figure 12 shown.

[0100] Example 9: Preparation of D. salina-curcumin by endocytosis

[0101] The difference from Example 1 is that quercetin is replaced by curcumin, and the mixture is co-incubated with the living cells of Dunaliella salina at 37° C., and the drug is loaded into the living cells of Dunaliella salina through endocytosis;

[0102] Figure 13 Drug loading rates at different concentrations of curcumin (9.375 μM, 18.75 μM, 37.5 μM, 75 μM, 150 μM, 300 μM, 600 μM).

[0103] Example 10: Preparation of D. salina-liquiritin by endocytosis

[0104] The difference from Example 1 is that quercetin is replaced by liquiritin, and then the mixture is co-incubated with the living cells of Dunaliella salina at 37° C., and the drug is loaded into the living cells of Dunaliella salina through endocytosis;

[0105] Figure 14 Drug loading rates at different concentrations of liquiritin (6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM, 200 μM, 400 μM).

[0106] In Example 9-10, quercetin was replaced with curcumin and liquiritin, and the endocytosis method was used to prepare a drug delivery carrier for Dunaliella cells. The results showed that different concentrations of curcumin and liquiritin could be effectively loaded into Dunaliella cells by this method, and the loading rate showed a regular change with the drug concentration, such as Figure 13-14This result confirms the broad applicability of the preparation method for drug delivery vehicles based on Dunaliella cells, breaking through the limitations of single drugs and providing a feasible solution for constructing efficient delivery systems for various natural anti-tumor drugs such as curcumin and liquiritin. It further verifies the universality and stability of this preparation method in different drug loading scenarios.

[0107] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a drug delivery carrier based on Dunaliella cells, characterized in that: The following steps are involved: S1: Dunaliella salina was selected for standardized culture to obtain living cells of Dunaliella salina; S2: The target drug and the living cells of Dunaliella are loaded into the living cells of Dunaliella through endocytosis; S3: Centrifuge to remove free drug molecules and obtain Dunaliella cell drug delivery carrier.

2. The method according to claim 1, characterized in that The step S1 specifically includes: S101: preparing modified Dunaliella culture medium; S102: In a conical flask, at 26°C and a light intensity of 50 μM photons·m -2 ·s -1 , culturing Dunaliella salina cells in a modified Dunaliella medium under a light intensity of 12 hours per day; S103: When the cells grew to the logarithmic phase, they were collected by centrifugation; S104: Wash the cells three times with fresh medium and adjust the cell concentration to 5 × 10 6 / mL; S105: Centrifuge the treated cell mixture at a rotation speed of 2500 rpm for 5 minutes to obtain live Dunaliella cells.

3. The method according to claim 2, characterized in that The modified Dunaliella culture medium in step S101 includes: 1.5M NaCl, 1μM CuCl·2H2O, 10mM KNO3, 50mM NaHCO3, 0.4mM KHPO4, 185μM H3BO3, 2μM FeCl6·H2O, 5mM MgSO4·7H2O, 5μM EDTA, 1μM (NH4)MO7O 24 ·4H2O, 7μM MnCl2·4H2O, 1μM ZnCl2, 1μM CoCl26·6H2O, and 0.2mM CaCl2.

4. The method according to claim 3, characterized in that In the logarithmic stage in step S103, the concentration of Dunaliella cells is 1×10 6 cells / mL to 1×10 7 pieces / mL.

5. The method according to claim 1, characterized in that The step S2 specifically includes: S201: Pretreatment of Dunaliella live cells a. Prepare a pretreatment solution containing 2.5 mM ATP, 2.5 mM MgCl2, 1 mM CaCl2; b. Incubate 1 volume of Dunaliella live cells with 9 volumes of the pretreatment solution at room temperature for 5 minutes; c. Centrifuge at 2500 rpm for 5 minutes to obtain the pretreated Dunaliella cells; S202: Target drug loading a. Dissolve the target drug in 154 mM NaCl solution; b. Incubate the Dunaliella cells obtained in step S201 with the target drug solution at 37°C for 10 minutes; c. Centrifuge at 2500 rpm for 5 minutes to obtain the Dunaliella cell drug delivery vector.

6. The method according to claim 5, characterized in that The target drug is quercetin, curcumin or liquiritin.

7. A drug delivery carrier prepared by the method according to claim 6.

8. Use of the Dunaliella cell drug delivery vector according to claim 7 in preparing a D. salina-QCT delivery vector.

9. Use of the Dunaliella cell drug delivery vector according to claim 7 in preparing a D. salina-curcumin delivery vector.

10. Use of the Dunaliella cell drug delivery vector according to claim 7 in preparing a delivery vector for D. salina-liquiritin.

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