Nucleic acid delivery carrier based on nano aluminum adjuvant and application of nucleic acid delivery carrier

Abstract

The invention discloses a nucleic acid delivery carrier based on a nano aluminum adjuvant and application thereof, the nucleic acid delivery carrier is composed of an inner core, a middle layer and a shell from inside to outside in sequence, the inner core is a compound of cationic protein and nucleic acid, the middle layer is a lactide-glycolide copolymer (PLGA), and the shell is the nano aluminum adjuvant. The cationic protein can be tightly combined with nucleic acid with negative charges through electrostatic interaction to form a stable compound, so that the nucleic acid is prevented from being degraded; pLGA is introduced as a middle layer, the stability of the inner core can be further enhanced, and leakage of nucleic acid in the delivery process is prevented; the nano-aluminum is used as the outermost layer to provide an additional physical barrier, so that the stability and the biological safety of the whole delivery carrier can be enhanced, and nucleic acid is ensured not to be degraded or inactivated before reaching a target cell; therefore, the carrier provided by the invention can realize delivery of high nucleic acid drug loading capacity.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a nucleic acid delivery vector based on nano-aluminum adjuvant and application thereof. Background Art

[0002] Vaccines, as biological preparations used to prevent and control infectious diseases, are usually composed of antigens (attenuated, inactivated pathogens or their subunit products) and adjuvants. They stimulate the body's immune system to produce corresponding antibodies and immune cells, thereby enabling the body to gain immunity to specific pathogens without causing disease. The widespread use of vaccines has enabled smallpox to be cured, and the incidence of many diseases such as polio, rabies, and typhoid fever has dropped significantly. However, traditional vaccines still face major obstacles in dealing with many major diseases, such as the treatment of acute infectious diseases, malignant tumors, and other diseases. mRNA nucleic acid vaccines have relevant advantages: mRNA, as a transient vector, does not need to enter the cell nucleus, has no transcription process, and can be directly translated in the cytoplasm with high translation efficiency; based on the characteristics of mRNA that it does not need to be inserted into the genome, there is no risk of gene mutation and high safety; at the same time, mRNA drugs have a short half-life, controllable expression time, and controllable side effects.

[0003] As the core technology of vaccines, adjuvants can improve the immunogenicity, stability, immune response type, vaccination frequency, dosage, etc. of vaccines by promoting targeted delivery, tissue penetration, cell uptake, controlled release and synergistic treatment of antigens. Currently, the FDA has approved several human vaccine adjuvants (aluminum adjuvant, MF59, AS04, AS03, AS01, CpG1018, Matrix-M), among which aluminum adjuvant is the most widely used and the only adjuvant approved for use in my country. At present, clinical studies on the potential of aluminum adjuvants to enhance immune responses include antigen reservoir effects, pro-phagocytosis, pro-inflammatory responses, and complement system activation.

[0004] Currently, clinical trials of mRNA vaccine products for advanced solid tumors (including lung cancer, melanoma, and pancreatic cancer) are being carried out globally. Anti-tumor mRNA vaccines mainly work through the absorption of mRNA by antigen-presenting cells and the activation of antigen-specific CD8+T cells. The initiation and activation of T cells are key links in the development of anti-tumor immunity. The proliferation, differentiation, and functional effects of T cells are mediated by signaling molecules on the surface of T cells. T cell surface signals can be divided into co-inhibitory molecules: PD1, CTLA4, etc., and co-stimulatory molecules include members of the tumor necrosis factor receptor superfamily (TNFRSF) or immunoglobulin superfamily (IgSF). TNFRSF members include GITR, OX40, 4-1BB, etc., and IgSF members include CD28, ICOS, etc. So far, a variety of T cell co-stimulatory molecules are in different stages of clinical research, showing pharmacodynamic activity and immunomodulatory effects.

[0005] At present, most clinical trials of anti-tumor mRNA vaccines are still in the early stages. Tumors are highly heterogeneous and dynamically mutated, which makes it difficult for mRNA vaccines targeting specific mutations to fully cover all tumor cells. In addition, PEGylated lipid delivery system LNPs are the mainstream carriers for mRNA delivery. LNPs are mainly composed of cationic lipids, auxiliary phospholipids, cholesterol and PEG-anchored lipids. The mRNA loading is less than 4%, and it is difficult to break through the dense barrier of solid tumors and achieve efficient infiltration of immune cells. Summary of the invention

[0006] One of the purposes of the present invention is to provide a nucleic acid delivery vector, which is composed of an inner core, an intermediate layer and an outer shell from the inside to the outside, wherein the inner core is a complex of a cationic protein and a nucleic acid, the intermediate layer is poly(lactide-co-glycolide) (PLGA), and the outer shell is nanoaluminum.

[0007] Furthermore, the cationic protein is protamine or ribosomal protein; the nucleic acid is mRNA, siRNA, DNA, microRNA or saRNA.

[0008] Furthermore, the molecular weight of the lactide-glycolide copolymer is 5000-60000, and the molar ratio of lactide (PLA) to glycolide (PGA) in the lactide-glycolide copolymer is 1:99-99:1.

[0009] Furthermore, the particle size of the nano aluminum is 0.01-1 μm.

[0010] Furthermore, the nano aluminum is AlOOH, Al(OH)3 or Al2O3·3H2O.

[0011] The second object of the present invention is to provide a method for preparing the above-mentioned nucleic acid delivery vector, comprising the following steps:

[0012] Step 1, mixing the nucleic acid and the cationic protein solution to obtain a cationic protein / nucleic acid complex;

[0013] Step 2, adding the cationic protein / nucleic acid complex to the lactide-co-glycolide copolymer solution, and ultrasonically emulsifying to obtain water-in-oil colostrum;

[0014] Step 3, adding water-in-oil colostrum to the nano-aluminum solution to obtain the delivery vehicle.

[0015] Furthermore, the nano-aluminum solution in step 3 contains a stabilizer, and the stabilizer is selected from polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP) or carboxymethyl cellulose (CMC).

[0016] The third object of the present invention is to provide the use of the above-mentioned nucleic acid delivery vector in the preparation of tumor therapeutic drugs.

[0017] In the present invention, a complex formed by cationic protein and nucleic acid is used as the core, and then an amphiphilic polymer is added to form an intermediate layer, and finally nano-aluminum is coated as the outermost layer to obtain a delivery carrier loaded with nucleic acid. Among them, cationic protein can be closely combined with negatively charged nucleic acid through electrostatic interaction to form a stable complex to protect nucleic acid from degradation; by adjusting the ratio of cationic protein to nucleic acid, the amount of nucleic acid loaded can be accurately controlled while maintaining the stability and biocompatibility of the complex. The introduction of lactide-co-glycolide copolymer as an intermediate layer can further enhance the stability of the core and prevent the leakage of nucleic acid during delivery; by changing the type, concentration and structure of amphiphilic polymers, the interaction between the carrier and the biomembrane can be improved, and by fine-tuning the physical properties of the carrier (such as size, shape, surface charge, etc.), it can adapt to the needs of different cell types and delivery conditions. Nano-aluminum provides an additional physical barrier as the outermost layer, which can enhance the stability and biosafety of the entire delivery carrier and ensure that the nucleic acid is not degraded or inactivated before reaching the target cell; at the same time, nano-aluminum has good biocompatibility, controllable degradability and clear adjuvant effect, which can enhance the immune response.

[0018] Beneficial effects:

[0019] (1) Compared with the traditional micron-sized aluminum adjuvant, the nano-sized aluminum adjuvant prepared by the present invention containing the same amount of aluminum ions has a smaller particle size, a larger specific surface area, and a higher surface reactivity; the nano-sized aluminum adjuvant can induce therapeutic CD8 + T cell-mediated immune response improves the efficacy of tumor vaccines.

[0020] (2) The materials used, protamine and PLGA, have been clinically approved. Aluminum adjuvant is the most widely used human adjuvant and the only adjuvant approved for use in my country. The double emulsion method has a simple preparation process, is easy to operate, and is easy to clinically transform.

[0021] (3) In the present invention, CD137L mRNA is selected as the target mRNA. CD137L is expressed on DC cells, and CD137 is expressed on T cells. CD137 / CD137L, as a pair of co-stimulatory molecules, plays an important role in the expansion, acquisition of effector function, survival and development of T cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is the particle size test result of nano aluminum.

[0023] Figure 2 This is the potential detection result of nano-aluminum.

[0024] Figure 3 This is the detection result of protamine-encapsulated mRNA.

[0025] Figure 4 The particle size and morphology of PA1 / mRNA microparticles are shown in Figure 1. The scale bar is 1 μm.

[0026] Figure 5 It is the potential detection result of P / mRNA and PA1 / mRNA.

[0027] Figure 6 This is the result of RNAse stability test of PA1 / mRNA.

[0028] Figure 7 The results of DC cell uptake of PA1 / mRNA are shown in Figure 2. Scale bar: 10 μm.

[0029] Figure 8 Shown are the DC cell transfection results of P / mRNA and PAl / mRNA.

[0030] Fig. 9 The results show the expression of CD137L mRNA in DC cells.

[0031] Fig.10 This is the result of CD137L mRNA promoting DC cell maturation. DETAILED DESCRIPTION

[0032] In the present invention, a complex formed by cationic protein and nucleic acid is used as the core, PLGA is then added to form an intermediate layer, and finally nano-aluminum is coated as the outermost layer to obtain a delivery carrier loaded with nucleic acid.

[0033] In one embodiment of the present invention, CD137L mRNA is used as a model mRNA, a complex of protamine and protamine is first used as a core, and then the protamine / mRNA complex is added to a PLGA solution for ultrasonic emulsification to obtain an oil-in-water colostrum, and finally the oil-in-water colostrum is added to a PVA solution containing nano-aluminum to prepare a delivery carrier. The mass ratio of protamine:mRNA is 1:10-20:1; the volume ratio of protamine / mRNA complex and PLGA solution is 1:10, and the concentration of PLGA solution is 5-200 mg / mL; the volume ratio of oil-in-water colostrum and PVA solution containing nano-aluminum is 1:5, the concentration of nano-aluminum is 1-50 mg / mL, and the concentration of PVA is 0.1-20%.

[0034] The preferred embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that the following examples are provided only for the purpose of illustration and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0035] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0036] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0037] In the following examples, EGFP mRNA and FAM-RNA were purchased from GenScript; CD137L mRNA sequence was determined by querying the gene library (Gene ID: 21950), constructing plasmids, and in vitro transcription. Poly(lactic-co-glycolic acid) (PLGA, Mw 38000-54000Da, LG 50:50).

[0038] Examples 1-6

[0039] 1. Preparation and characterization of nano-aluminum

[0040] 1.1 Preparation of aluminum oxyhydroxide (AlOOH)

[0041] The synthesis method of nano-scale AlOOH is as follows: 30 mL of 0.04 mol / L NaOH solution is added dropwise to 30 mL of 3.6 mg / mL AlCl3·6H2O aqueous solution, and then the pH is adjusted to 5.20 with 0.01 mol / L NaOH. Ultrasonication is performed for 30 min in an ice bath with an ultrasonic crusher, and the ultrasonic power is set to 300 W. The solution is placed in a 50 mL ultrafiltration tube (MWCO 3 kDa), centrifuged at 5000 rpm, and residues are removed to obtain a nano-aluminum concentrate.

[0042] The particle size distribution and potential of nano-aluminum were evaluated by dynamic light scattering particle size analyzer. Figure 1 and Figure 2 As shown, the particle size of nano-aluminum is 57.87nm, and it shows obvious Tyndall effect under laser irradiation, and the potential of nano-aluminum is +42.8mV.

[0043] 1.2 Preparation of aluminum hydroxide (Al(OH)3)

[0044] Weigh 32g AlCl3 and dissolve it in 40mL deionized water to obtain 80% AlCl3 solution. Weigh 16g NaOH and dissolve it in 40mL deionized water to obtain 40% NaOH solution. Add AlCl3 solution into a reaction vessel and stir at room temperature at a stirring speed of 150-200r / min, and add NaOH solution dropwise into AlCl3 solution until pH reaches 5.8 to obtain Al(OH)3 solution.

[0045] 1.3 Preparation of hydrated aluminum oxide (Al2O3·3H2O)

[0046] Take 32g of anhydrous AlCl3 crystals and put them into a three-necked flask. Add a certain amount of benzene to moisten it. Then slowly add an appropriate amount of anhydrous ethanol. AlCl3 is completely dissolved into a transparent solution. Keep the temperature at 60-70℃. Then stir and pass ammonia to promote the reaction. The reaction is continued for 2h. The white solid in the reflux reactor gradually increases, and the reaction stops when the pH is 8. Then vacuum filter and distill the filtrate in a water bath. Take the distilled sample and dissolve it in deionized water for hydrolysis to obtain Al2O3·3H2O.

[0047] 2. Preparation and characterization of PA1 / mRNA vaccine

[0048] Use RNase-free DEPC water to prepare an mRNA solution with a concentration of 80 ng / μL, and use 10 mM HEPES buffer (pH 7.4) to prepare protamine solutions with concentrations of 16, 26.7, 40, 80, 160, 240, 400, and 800 ng / μL, respectively. Take 5 μL of each and mix them in equal volumes to form 10 μL mixed solutions with a protamine / mRNA mass ratio of w / w = 1:5, 1:3, 1:2, 1:1, 2:1, 3:1, 5:1, and 10:1, respectively. 10 μL of the mixed solution was mixed with 2 μL of RNA loading buffer, and electrophoresis analysis was performed using 1% agarose gel stained with GelRed (120V, 10 min). The exposed mRNA bands were observed and photographed by UV development. The results are shown in Figure 3As shown, when the protamine / mRNA mass ratio was 1:1 or above, protamine could completely compress mRNA, so a w / w ratio of 1:1 was selected for subsequent experiments.

[0049] 15 μL of 2 μg / μL CD137L mRNA solution and 15 μL of 2 μg / μL protamine solution were mixed to form a protamine / mRNA complex, with the mass ratio of protamine to CD137L mRNA being w / w=1:1.

[0050] Press V 水相 :V 油相 =1:10, the protamine / mRNA complex was added as the core to 300 μL of PLGA dichloromethane solution, the power was set to 80W, the time was set to 1min, and ultrasonic emulsification was performed to form oil-in-water (W / O) colostrum. Then, the oil-in-water (W / O) colostrum was added to 1.5mL of 1% PVA aqueous solution containing nano-aluminum, magnetic stirring was performed, and the organic solution was volatilized to obtain a water-in-oil-in-water (W / O / W) emulsion, and PAl / mRNA was obtained at this time.

[0051]

[0052] It can be seen from the above table that when the PLGA concentration is too high, the particle size of the prepared emulsion is larger. In order to control the particle size of the emulsion, 25 mg / mL of PLGA was selected in subsequent experiments; in order to ensure the adsorption of nano-aluminum and the subsequent effect of promoting DC maturation, 10 mg / mL of nano-aluminum was selected in subsequent experiments.

[0053] 15 μL of 2 μg / μL CD137L mRNA solution and 15 μL of 2 μg / μL protamine solution were mixed to form a protamine / mRNA complex as the core, and the mass ratio of protamine to CD137L mRNA w / w=1:1. The protamine / mRNA complex was added dropwise to 300 μL of 25 mg / mL PLGA solution (the volume ratio was kept at V 水相 :V 油相 =1:10), set the power to 80W, set the time to 1min, perform ultrasonic emulsification to form water-in-oil (W / O) colostrum. Add the water-in-oil (W / O) colostrum dropwise to 1.5mL of 1% PVA aqueous solution containing nano-aluminum (10.0mg / mL AlOOH), stir magnetically, volatilize the organic solution, and obtain water-in-oil-in-water (W / O / W) emulsion, i.e., PLGA microparticles (PAl / mRNA) coated with nano-aluminum adjuvant and loaded with mRNA.

[0054] In addition, water-in-oil (W / O) colostrum was added dropwise to 1.5 mL of 1% PVA aqueous solution without nano-aluminum, and magnetic stirring was performed to evaporate the organic solution to obtain water-in-oil-in-water (W / O / W) emulsion, which is PLGA microparticles encapsulating mRNA (P / mRNA).

[0055] The particle size distribution and potential of P / mRNA and PAl / mRNA were evaluated by dynamic light scattering particle size analyzer, and the particle morphology was observed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Figure 4 and Figure 5 As shown, the particle size of PAl / mRNA is 1053nm (PDI=0.233), the potential is +16.9mV, while the potential of P / mRNA is -1.99mV. The increase of the potential of PAl / mRNA is due to the coating of the outer layer of nano-aluminum.

[0056] Comparative Example 1

[0057] Preparation of blank PLGA microparticles

[0058] Press V 水相 :V 油相 =1:10, 30 μL of aqueous solution was added dropwise to 300 μL of 25 mg / mL PLGA solution, the power was set to 80 W, the time was set to 1 min, and ultrasonic emulsification was performed to form water-in-oil (W / O) colostrum. Then, the water-in-oil (W / O) colostrum was added dropwise to 1.5 mL of 1% PVA aqueous solution, magnetic stirring was performed, and the organic solution was volatilized to obtain a water-in-oil-in-water (W / O / W) emulsion, which was blank PLGA particles.

[0059] In addition, water-in-oil (W / O) colostrum was added dropwise to 1.5 mL of 1% PVA aqueous solution containing nanoaluminum (10.0 mg / mL AlOOH), and magnetic stirring was performed to volatilize the organic solution to obtain a water-in-oil-in-water (W / O / W) emulsion, which is PLGA microparticles (PAl) coated with nanoaluminum adjuvant.

[0060] Test Example 1

[0061] Detection of mRNA integrity after incubation of PA1 / mRNA with RNase

[0062] PAl / mRNA was prepared by the method in Example 1, the mRNA was CD137L mRNA, and the mass ratio of protamine to mRNA was w / w=1:1. PAl / mRNA microparticles were incubated with RNase (2 μL, 5 U / μL) for different time periods (0, 2, 4, 8, 12, 24 h), and then 2% SDS solution was added and allowed to stand for 0.5-1 h to obtain free mRNA. Free mRNA was detected by agarose gel electrophoresis, and the gel device was placed in an ice water bath to prevent mRNA degradation due to high temperature. The results are shown in Figure 6 As shown, the brightness of the mRNA bands at different time points is the same as that of untreated mRNA (Naked mRNA), proving that PA1 / mRNA microparticles can protect mRNA from degradation in the presence of RNase.

[0063] Test Example 2

[0064] Evaluation of PA1 / mRNA uptake by DC2.4 cells using confocal laser scanning microscopy

[0065] PAl / mRNA was prepared by the method in Example 1, wherein the mRNA was green fluorescent labeled FAM-RNA, and the mass ratio of protamine to mRNA was w / w=1:1. DC2.4 cells were inoculated in a confocal culture dish and cultured at 37°C for 24 hours. PAl / mRNA microparticles (2 μg / mL mRNA) were added to the culture dish and incubated with DC2.4 cells for 6 hours. The supernatant was then discarded, and the cells were fixed with 4% paraformaldehyde for 15 minutes and stained with DAPI for 15 minutes. A laser confocal microscope was then used to observe the cell uptake. The results are shown in Figure 2. Figure 7 As shown, green fluorescence represents PAl / mRNA and blue fluorescence represents cell nuclei, and the results indicate that PAl / mRNA can be effectively taken up by DC2.4 cells.

[0066] Test Example 3

[0067] Evaluation of PA1 / mRNA transfection efficacy using an inverted fluorescence microscope

[0068] P / mRNA and PAl / mRNA were prepared by the method in Example 1, wherein the mRNA was EGFP mRNA expressing green fluorescent protein, and the mass ratio of protamine to mRNA was w / w=1:1. DC2.4 cells were inoculated in a 6-well plate and cultured at 37°C for 24 h. PAl / mRNA microparticles (2 μg / mL mRNA) were added to serum-free medium and incubated with DC2.4 cells for 6 h, after which the supernatant was discarded and replaced with serum-containing medium for 48 h. The green fluorescent protein expression of the cells was observed using an inverted fluorescence microscope, and the results were as follows: Figure 8 As shown, P / mRNA and PA1 / mRNA can promote the expression of green fluorescent protein in DC cells.

[0069] Test Example 4

[0070] Flow cytometry was used to detect the expression of CD137L in DC cells

[0071] P / mRNA and PAl / mRNA were prepared by the method in Example 1, wherein the mRNA was CD137L mRNA, and the mass ratio of protamine to mRNA was w / w=1:1. DC2.4 cells were inoculated in 6-well plates and cultured at 37°C for 24 hours. OVA (O), OVA and blank PLGA microparticles (O+P), OVA and PLGA microparticles coated with nanoaluminum adjuvant (O+PAl), OVA and PLGA microparticles loaded with mCD137L (O+P / mCD137L, 2μg / mL mRNA), OVA and PLGA microparticles coated with nanoaluminum adjuvant and loaded with mCD137L (O+PAl / mCD137L, 2μg / mL mRNA) were added to the serum-free medium and incubated with DC2.4 cells for 6 hours, after which the supernatant was discarded and replaced with serum-containing medium for incubation for 48 hours. The expression of CD137L in the cells was detected by flow cytometry, and the results were as follows: Fig. 9 As shown, P / mRNA and PA1 / mRNA can promote the expression of CD137L molecules on the surface of DC cell membrane.

[0072] Test Example 5

[0073] Flow cytometry was used to detect the maturation of DC cells in vitro

[0074] P / mRNA and PAl / mRNA were prepared by the method in Example 1, wherein the mRNA was CD137L mRNA, and the mass ratio of protamine to mRNA was w / w=1:1. DC2.4 cells were inoculated in 6-well plates and cultured at 37°C for 24h. OVA (O), OVA and blank PLGA microparticles (O+P), OVA and PLGA microparticles coated with nanoaluminum adjuvant (O+PAl), OVA and PLGA microparticles loaded with mCD137L (O+P / mCD137L, 2μg / mL mRNA), OVA and PLGA microparticles coated with nanoaluminum adjuvant and loaded with mCD137L (O+PAl / mCD137L, 2μg / mL mRNA) were added to the serum-free medium and incubated with DC2.4 cells for 6h, after which the supernatant was discarded and replaced with serum-containing medium for incubation for 48h. The expression of CD80 and CD86 in DC cells was detected by flow cytometry, and the results were as follows: Fig.10 As shown, the O group and the O+P group had no promoting effect on DC maturation, the O+PA1 group promoted DC maturation by about 2.3 times, the O+P / mCD137L group promoted DC maturation by about 1.4 times, and the O+PA1 / mCD137L group promoted DC maturation by about 4.1 times.

Claims

1. A nucleic acid delivery vector, characterized in that The nucleic acid delivery carrier is composed of an inner core, an intermediate layer and an outer shell from the inside to the outside, wherein the inner core is a complex of cationic protein and nucleic acid, the intermediate layer is a lactide-glycolide copolymer, and the outer shell is nano-aluminum.

2. The nucleic acid delivery vector according to claim 1, characterized in that The cationic protein is protamine or ribosomal protein; the nucleic acid is mRNA, siRNA, DNA, microRNA or saRNA.

3. The nucleic acid delivery vector according to claim 1, characterized in that The molecular weight of the lactide-glycolide copolymer is 5000-60000, and the molar ratio of lactide to glycolide in the lactide-glycolide copolymer is 1:99-99:

1.

4. The nucleic acid delivery vector according to claim 1, characterized in that The particle size of the nano aluminum is 0.01-1 μm.

5. The nucleic acid delivery vector according to claim 4, characterized in that The nano aluminum is AlOOH, Al(OH)3 or Al2O3·3H2O.

6. The method for preparing the nucleic acid delivery vector according to claim 1, characterized in that: The following steps are involved: Step 1, mixing the nucleic acid and the cationic protein solution to obtain a cationic protein / nucleic acid complex; Step 2, adding the cationic protein / nucleic acid complex to the lactide-co-glycolide copolymer solution, and ultrasonically emulsifying to obtain water-in-oil colostrum; Step 3, adding water-in-oil colostrum to the nano-aluminum solution to obtain the delivery vehicle.

7. The preparation method according to claim 6, characterized in that: The nano-aluminum solution in step 3 contains a stabilizer, and the stabilizer is selected from polyvinyl alcohol, polyvinyl pyrrolidone or carboxymethyl cellulose.

8. Use of the nucleic acid delivery vector according to any one of claims 1 to 5 in the preparation of tumor therapeutic drugs.

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