阅读说明：本技术 甘露糖受体靶向的组合物、药物及其制备方法和应用 (Composition, the drug and its preparation method and application of mannose receptor targeting ) 是由 宋相容 魏于全 于 2019-05-13 设计创作，主要内容包括：本发明属于药物制剂领域,具体涉及一种带有靶向功能的纳米制剂的组合物、靶向载体及其制备方法、靶向药物和提高靶向载体的靶向性的方法。所述带有靶向功能的纳米制剂的组合物所述靶向材料、所述基础纳米制剂材料、使所述靶向材料与所述基础纳米制剂材料产生距离的间隔材料组成。所述靶向纳米制剂具较好的甘露糖受体的靶向性,可以高效的与靶细胞上的甘露糖受体结合,且制备方法具有通用性,可以合成多种靶向纳米制剂,并有利于纯化和表征。(The invention belongs to field of pharmaceutical preparations, and in particular to a method of the composition of the nanometer formulation with target function, targeting vector and preparation method thereof, targeted drug and the targeting for improving targeting vector.Targeting material described in the composition of the nanometer formulation with target function, the basic nanometer formulation material, the interval insulant for making the targeting material and the basic nanometer formulation material generate distance form.The targeted nano preparation has the targeting of preferable mannose receptor, can be efficiently in conjunction with the mannose receptor on target cell, and preparation method has versatility, can synthesize a variety of targeted nano preparations, and is conducive to purify and characterize.)

1. being used to prepare the composition of the nanometer formulation with target function, which is characterized in that the composition includes targeting material Material and basic nanometer formulation material, the targeting material are mannose and/or mannose derivative.

2. composition according to claim 1, which is characterized in that the mannose derivative is mannoside and/or sweet Reveal osamine and/or mannosan.

3. composition according to claim 2, which is characterized in that the mannose derivative be methyl-Dmannose glycosides, 1- α formyl methyl-mannopyrane glucosides, 4- aminophenyl-α-D- mannopyranose glycosides, 4- nitrobenzophenone-α-D- pyrans sweet dew Glucosides, 4-methyl umbelliferone base-α-D- mannopyranose glycosides, Man-6-P, carbamoyl-D-MANNOSE.

4. composition according to claim 1 or 2 or 3, which is characterized in that the targeting material accounts for the weight of the composition Amount percentage is 0.05-40%.

5. composition according to claim 1 or 2 or 3, which is characterized in that the targeting material accounts for the weight of the composition Amount percentage is 1-10%.

6. composition according to claim 1, which is characterized in that the composition is by the targeting material, the basis Nanometer formulation material, the interval insulant for making the targeting material and the basic nanometer formulation material generate distance form, described Interval insulant is glutaraldehyde, ethylenediamine, malonic acid, short-chain alkyl chain, PEG, amino acid, dipeptides, oligopeptides, polypeptide, stearoyl, palm fibre One or more of palmitic acid acyl.

7. composition according to claim 1, which is characterized in that it is described basis nanometer formulation material be prepare liposome, Emulsion, nanogel, core-shell type nano grain, HDL nanoparticle, solid rouge nanoparticle, polymer micelle material composition.

8. composition according to claim 6, which is characterized in that the interval insulant is PEGn, wherein n=100- 5000。

9. composition according to claim 8, which is characterized in that the n be 100,200,300,400,500,600, 800,900,1000 or 2000.

10. composition according to claim 6, which is characterized in that composed of the following components in parts by weight: mannose 1 part；0.5-56 parts of PEG, 2-11 parts of cholesterol.

11. composition according to claim 6, which is characterized in that composed of the following components in parts by weight: mannose 1 part；10-25 parts of PEG, 4-8 parts of cholesterol.

12. the targeting vector of the drug delivery of composition preparation described in claim 1.

13. targeting vector according to claim 12, which is characterized in that the drug is small-molecule drug and/or albumen Polypeptide drug and/or genomic medicine.

14. targeting vector according to claim 13, which is characterized in that the small-molecule drug is taxanes, camplotheca acuminata Bases, vinca, adriamycin, Epi-ADM, daunorubicin, epirubicin, anphotericin, methotrexate (MTX), cytarabine, 5- Fluorouracil, mitoxantrone or derivatives thereof, Gefitinib, oscapine, cis-platinum, carboplatin, oxaliplatin, Carmustine and Mongolian oak Pi Suzhong it is any in or it is a variety of.

15. targeting vector according to claim 13, which is characterized in that the protein and peptide drugs are leucocyte Jie Element, growth factor, interferon, integrin, monoclonal antibody, enzyme and insulin.

16. targeting vector according to claim 15, which is characterized in that the interleukins includes but is not limited to IL- 1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6, the growth factor include but is not limited to fibroblast growth factor, Liver growth factor, vascular endothelial growth factor, hemopoieticgrowth factor, the interferon include but is not limited to IFN α, IFN β, IFN γ, the tumor necrosis factor include but is not limited to TNF α, TNF β.

17. targeting vector according to claim 13, which is characterized in that the genomic medicine be DNA, plasmid, mRNA, One or more of siRNA, shRNA, microRNA.

18. the drug wrapped up by the targeting vector described in claim 12.

19. the preparation method of targeting vector described in claim 12, it is characterised in that: by the basic nanometer formulation material system It is standby to transport drug carrier at nanometer, then the targeting material is invested into the nanometer and transports drug carrier surface.

20. the method for improving the targeting of targeting vector described in claim 12, it is characterised in that:

A synthesis targeting element

The targeting material is synthesized to targeting element with the interval insulant；

B prepares targeting vector

Basic nanometer formulation material is prepared into nanometer fortune drug carrier, then the resulting targeting element of step A and the nanometer are transported Drug carrier connects to obtain targeting vector；Or first by the interval insulant and basic nanometer formulation materials synthesis, then with the targeting material Material is further connected to obtain targeting vector.

21. according to the method for claim 20, which is characterized in that the targeting material is specially mannose and its derivative Object, the interval insulant are specially PEG.

22. according to the method for claim 20, it is characterised in that: it is described targeting element and the nanometer fortune drug carrier away from From for 0.2-100nm.

23. according to the method for claim 22, it is characterised in that: it is described targeting element and the nanometer fortune drug carrier away from From for 0.3-10nm.

24. transporting the method that effective component reaches target cell, it is characterised in that: effective component is placed in the targeting vector, And it transports.

25. according to the method for claim 20, feature is: the target cell is containing the thin of mannose receptor Born of the same parents.

26. according to the method for claim 20, it is characterised in that: the target cell includes but is not limited to macrophage, tree Prominent cell and tumour cell.

Technical field

The invention belongs to field of pharmaceutical preparations, and in particular to a kind of composition, the target of the nanometer formulation with target function To the method for the targeting of carrier and preparation method thereof, targeted drug and raising targeting vector.

Background technique

In recent years, the targeted drug delivery system mediated based on drug receptor has been widely studied, targeted drug transmitting system Its specific target tropism can be improved in system, overcomes the problems, such as that drug distribution is wide, toxic side effect is serious.Wherein, agglutinin receptor is One kind is distributed in the glycoprotein, glycolipid or saccharide complex of cell membrane surface, can specific recognition and bound fraction glycosyl.Mannose Receptor is most important efficient endocytosis agglutinin receptor, and function includes removing endogenous molecule, promoting antigen presentation, adjust carefully Born of the same parents' activation and transport, also with the immunologic escape of tumour and shift closely related, be mainly expressed in macrophage, dendritic cells with Tumour cell, can specific recognition mannose glycosyl molecule.Mannose group as most potential targeting group, have it is nontoxic, It many advantages, such as non-immunogenicity, good biocompatibility and biodegradability, can be widely used for drug delivery system It is glycosylation modified.

As most classic nano target drug administration carrier, the exposure basis of targeting is based on difference for liposome, nanoparticle Organ is selective to diameter of particle size intake situation difference to be gathered in the lymphoid tissues organ such as lung, to improve anti-swollen The therapeutic effect in each organ of tumor medicine.It, which is prepared, simply has controlled release, non-immunogenicity, and the features such as raising curative effect. But after targeting is realized in the endocytosis of liposome, nanoparticle dependence macrophage, stabilization during targeting dilivery Property and lesion part intake efficiency it is also relatively low, targeting also needs further continuous improve.Thus with life The continuous development of object pharmacy, material science, nanotechnology improves nanoparticle targeting and novel targeting vector exploitation gradually As the research hotspot in antitumor field, occurs a variety of various forms of nano targets at this stage and pass medicine type.

For using liposome as the targeting vector of drug, according to the principle that its targeting plays, ratio is studied to obtain at present More mature is divided into physics targeting and molecular target tropism liposome.Wherein physics targeted hposome includes pH sensitive liposome Body (PLP), photosensitive liposomes, magnetic liposome, thermo-responsive liposome etc..By taking photosensitivity liposome as an example, the benefits such as Yang Peptide pcCPP is permeated with light sensitive cells and contains the short of aspartic-glycine-arginine residues (Asn-Gly-Arg, NGR) Peptide is connected to surface of liposome, constructs pcCPP-NGR-LP, which can promote cellular uptake, and effectively sink Silent c-mycc gene, delays the growth of human fibrosarcoma.However, although above various forms of liposomes add to a certain extent Strong targeting, but still have shortcoming, pH sensitive liposome not can solve the liver and spleen cumulative toxicity problem of liposome.It is photosensitive The problems such as feasible and storage ability discharged with magnetic liposome organism, is still urgently to be resolved.Thermal sensitive liposome can be straight Connect killing tumor cell, but heating time too long causes normal tissue injury.There is also drugs for molecular target tropism liposome Orientation conveying and internal target organ absorption it is bad the problems such as.

To sum up, targeted drug is researched and developed, primarily solves the problems, such as to be exactly that nano-carrier keeps it steady during transportation Determine without being destroyed, (some there may be passive target due to not having active targeting for common nanoparticle or liposome Property), it is easily removed by endothelial system, reduces utilization rate, it is also possible to cause unnecessary toxicity.It secondly is exactly that drug reaches After target cell, by more effective ligand, more rapidly and directly enter the key point that cells play drug effect is research.

Summary of the invention

It, should one of the objects of the present invention is to provide being used to prepare with the selectively targeted material compositions for carrying drug carrier Composition can be used in preparing the strong load drug carrier of targeting.

To achieve the above object, the technical solution of the present invention is as follows:

It is used to prepare the composition of the nanometer formulation with target function, the composition includes that targeting material and basis are received Metric system agent material, the targeting material are mannose and/or mannose derivative.

Further, the mannose derivative is mannoside and/or mannosamine and/or mannosan.

Further, the mannose derivative be methyl-Dmannose glycosides, 1- α formyl methyl-mannopyrane glucosides, 4- aminophenyl-α-D- mannopyranose glycosides, 4- nitrobenzophenone-α-D- mannopyranose glycosides, 4-methyl umbelliferone base-α-D- pyrrole It mutters mannoside, Man-6-P, carbamoyl-D-MANNOSE.

Further, the weight percent that the targeting material accounts for the composition is 0.05-40%.

Preferably, the weight percent that the targeting material accounts for the composition is 1-10%.

Further, the composition by the targeting material, the basic nanometer formulation material, make the targeting material and The basis nanometer formulation material generates the interval insulant composition of distance, and the interval insulant is glutaraldehyde, ethylenediamine, the third two One or more of acid, short-chain alkyl chain, PEG, amino acid, dipeptides, oligopeptides, polypeptide, stearoyl, palmityl.

Specifically, the interval base be glutaraldehyde, ethylenediamine, malonic acid, short-chain alkyl chain (such as-CH2-CH2-), One or more of PEG, amino acid, dipeptides, oligopeptides, polypeptide etc..Specifically, interval base amino acid is arginine, asparagus fern acyl Amine, aspartic acid, glutamic acid, glutamine, lysine, serine, threonine, tyrosine.The both ends of dipeptides, few skin, polypeptide Amino acid amino acid composition selected from the following: arginine, asparagine, aspartic acid, glutamic acid, glutamine, lysine, silk Propylhomoserin, threonine, tyrosine；Central amino acid is selected from 20 kinds of arbitrary amino acids (such as RGD).

Further, the basic nanometer formulation material be prepare liposome, emulsion, nanogel, core-shell type nano grain, HDL nanoparticle, solid rouge nanoparticle, polymer micelle material composition.

Specifically, the basis nanometer formulation material is the composition for preparing nanometer formulation material, it can be " modern medicines Preparation technique " in the nanometer formulation material mentioned.

Specifically, the basis nanometer formulation material is lipid, poly lactide-glycolide acid (PLGA), polylactic acid (PLA), polycaprolactone (PCL), poly-D-lysine (PLL), polyethyleneimine (PEI), hyaluronic acid (HA), chitosan, gelatin, One or more of poloxamer, stearyl alcohol.

Specifically, liposome is prepared, the matrix material of core-shell type nano grain is cholesterol, egg yolk lecithin, soybean lecithin Rouge, cephalin, sphingomyelins, PC (phosphatidyl choline), EPG (phosphatidylglycerol), SPG (soy phosphatidylglycerol), distearyl Acyl phosphatidyl-ethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dipalmitoylphosphatidylcholine (DPPC), two oil Phosphatidyl choline (DOPC), Distearoyl Phosphatidylcholine (DSPC), dimyristoylphosphatidylcholine (DMPC), two sub- oleoyls Phosphatidyl choline (DLPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), two nutmeg acyl phosphorus Phosphatidylcholine (DMPC), dilauroylphosphatidylglycerol (DLPG), cetyl trimethylammonium bromide (CTAB), dimethyl double ten One of eight alkyl bromination ammoniums (DDAB), 1,2-dioleoyloxy-3-trimethylammonio propane (chloride salt) (DOTAP) are more Kind.

Specifically, the solid lipid material for preparing solid rouge nanoparticle be stearic acid, it is cholesterol, glycerin monostearate, double Tristerin, glyceryl tristearate, three Yu acid glycerides, glyceryl laurate ester, glycerol palmitic, stearic rouge, 20 Diacid monoglyceride, behenic acid double glyceride, behenic acid glyceryl ester, three myristins, citron acid glycerol One or more of ester, octadecyl alcolol, palmitinic acid, myristic acid, three Yu acid, lauric acid.

Further, the interval insulant is PEGn, wherein n=100-5000.

Preferably, the n is 200,400,1000 or 2000.

Further, composed of the following components in parts by weight: 1 part of mannose；0.5-56 parts of PEG, cholesterol 2-11 Part.

Preferably, composed of the following components in parts by weight: 1 part of mannose；10-25 parts of PEG, 4-8 parts of cholesterol.

Further, according to composed of the following components according to molar part meter: 1 part of mannose；1-5 parts of PEG, cholesterol 1-5 Part.

Preferably, composed of the following components according to molar part meter: 1 part of mannose；1-3 parts of PEG, 1-3 parts of cholesterol.

The second object of the present invention, which is to provide a kind of targeting vector and targeted drug, the targeting vector, can transport a variety of medicines It imitates ingredient and reaches target cell, and targeting is strong.

To achieve the above object, the present invention uses following scheme:

The targeting vector of the drug delivery of the composition preparation.

Further, the drug is small-molecule drug and/or protein and peptide drugs and/or genomic medicine.

The small-molecule drug be can be wrapped in nanometer formulation for neoplasm targeted therapy, artery sclerosis treatment and it is various Treatment the effect of property small molecule of diseases associated with inflammation etc..

In some embodiments, the mRNA for encoding particular tumor antigens can be wrapped in targeted nano preparations carrier, then is led Enter in body cell, and pass through the expression system synthetic antigen albumen of host cell, induction host immune system is generated to the tumour The immune response of antigen can realize the function of anti-curing oncoma, and the targeted nano preparation for being loaded with mRNA has better cancer target Property, be conducive to the treatment of tumour.Equally by antitumor chemical drug and biological medicament, the drug for the treatment of artery sclerosis and diseases associated with inflammation It can also be wrapped in targeted nano preparations carrier, play the role of targeted therapy more accurately.

Specifically, the small-molecule drug includes but is not limited to PTX, DOX, Quercetin, the protein and peptide drugs packet Albumin is included but is not limited to, the genomic medicine includes but is not limited to mRNA, siRNA.

More specifically, the small-molecule drug is that can be wrapped in nanometer formulation for neoplasm targeted therapy, artery sclerosis Treatment the effect of property small molecule for the treatment of and various diseases associated with inflammation etc..Such as anti-tumor drug taxanes (taxol, mostly west Taxol, Cephalomannine, 7- table are to taxol), camptothecin (camptothecine, SN38, Irinotecan, 9-aminocamptothecin, 9- Nitrocamptothecin etc.), vinca (vincaleukoblastinum, vincristine, eldisine, Vinorelbine, vinflunine etc.), adriamycin, Epi-ADM, daunorubicin, epirubicin, anphotericin, methotrexate (MTX), cytarabine, 5 FU 5 fluorouracil, mitoxantrone or its Derivative, Gefitinib, oscapine, cis-platinum, carboplatin, oxaliplatin, Carmustine, Quercetin etc..The main ingredient ingredient egg White polypeptide drug is interleukins (such as IL-1, IL-1a, IL-2, IL-3, IL-4, IL-5, IL-6 etc.), various growths The factor (such as fibroblast growth factor, liver growth factor, vascular endothelial growth factor, hemopoieticgrowth factor etc.), interference Plain (such as IFN α, IFN β, IFN γ), tumor necrosis factor (such as TNF α, TNF β), integrin, monoclonal antibody, enzyme, pancreas islet Element etc..The main ingredient component gene drug is DNA, plasmid, mRNA, siRNA, shRNA, microRNA etc..When targeted nano system When agent is used for immunization therapy, the main ingredient ingredient can also wrap up simultaneously immunologic adjuvant to increase in addition to the drug for immunization therapy Strong immune effect.

The drug wrapped up by the targeting vector.

Further, the drug is small-molecule drug and/or protein and peptide drugs and/or genomic medicine.

Specifically, the small-molecule drug includes but is not limited to PTX, DOX, Quercetin, the protein and peptide drugs packet Albumin is included but is not limited to, the genomic medicine includes but is not limited to mRNA, siRNA.

Further, the drug can be any pharmaceutically acceptable dosage form.

Specifically, the one or more pharmaceutically acceptable carriers of the pharmaceutically acceptable dosage form packet, diluent or figuration Agent and the addition of suitable step in above-mentioned preparation process.Term " pharmaceutically acceptable " used in the present invention refers in this way Compound, raw material, composition and/or dosage form, they within the scope of reasonable medical judgment, suitable for being contacted with patient tissue And without excessive toxicity, irritation, allergy or other problems relative to a reasonable benefit/risk ratio and complication, and Effective for given application.

Pharmaceutical preparation is suitable for being administered by any suitable approach, for example, by take orally (including oral cavity or sublingual), rectum, Nose, part (including oral cavity, sublingual or percutaneous), vagina or parenteral (including subcutaneous, intradermal, intramuscular, intra-articular, intrasynovial, chest Bone is interior, intrathecal, intralesional, intravenous or intradermal injection or infusion) approach.It can be by any known method of art of pharmacy This kind of preparation is prepared, such as by mixing active constituent with carrier or excipient.It is preferred that oral administration, local administration or injection Administration.Pharmaceutical preparation suitable for oral administration is provided by independent unit, for example, solution in aqueous or non-aqueous liquid or Suspension；Capsule or tablet；Powder or granule；Edible foam formulations or foaming preparations etc..

The third object of the present invention is to provide a kind of preparation method of targeting vector, and this method can be used for industrialized Prepare targeting vector.

To achieve the above object, the present invention uses following scheme:

The method of the preparation targeting vector are as follows: the basic nanometer formulation material is prepared into nanometer fortune drug carrier, The targeting material is invested into the nanometer again and transports drug carrier surface.

The method that the targeting material is invested nanometer fortune drug carrier surface includes but is not limited to that solvent is used to volatilize Method, film dispersion method, ultrasonic dispersion, injection method, reverse evaporation, high pressure homogenization method and micro emulsion method prepare Nano medication load After body, then will to target material solution mixed, makes to target material and invests its surface.It is above-mentioned prepare nano-medicament carrier method can Referring to " modern medicines preparation technique "

Specifically, the solvent evaporation method is a kind of method for preparing microballoon, simple process is exactly that first configuration is suitable dense Then the Polymer Solution of degree emulsifies Polymer Solution in continuous phase, gradually evaporation of solvent, be made microball preparation, then to Targeting material is added in the microball preparation, so that it is targeted material and is adsorbed on microball preparation surface.

High pressure homogenization method is that material passes through homogenizing valve under high pressure, is sprayed with high flow velocity, hits collision ring, passes through sky Cave, shock, shearing effect make material micronization and dispersion and emulsion, form nanoparticle, and the surface area of material increases during being somebody's turn to do Greatly, after targeting material being added thereto, the targeting material can invest surface.

Film evaporation method is that phospholipid material is dissolved in organic solvent, and then under a reduced pressure, organic solvent is steamed Hair, after forming a film on bottle inner wall, adding water or PBS solution will stir repeatedly, wash lower film through homogeneous, ultrasound at Reason is to get Nanoparticulate formulations, and after targeting material is added thereto, the targeting material can invest surface.

The fourth object of the present invention is to provide a kind of method for improving targeting vector targeting, and this method is obviously improved Targeting vector targeting.

To achieve the above object, the present invention uses following scheme:

The method for improving the targeting of the targeting vector, comprising the following steps:

A synthesis targeting element

The targeting material is synthesized to targeting element with the interval insulant；

B prepares targeting vector

Basic nanometer formulation material is prepared into nanometer formulation, then the resulting targeting element of step A and the nanometer are transported Drug carrier connects to obtain targeting vector or first by the interval insulant and basic nanometer formulation materials synthesis, then with the targeting material Further connected to obtain targeting vector.

Further, the targeting element is 0.2-100nm at a distance from nanometer fortune drug carrier.

Preferably, the targeting element is 0.3-10nm at a distance from nanometer fortune drug carrier.

The fifth object of the present invention is to provide a kind of method for transporting drug, the efficient drug delivery of this method energy to mesh Ground.

To achieve the above object, the present invention uses following scheme:

Transport the method that effective component reaches target cell are as follows: effective component is placed in the targeting vector, and is transported.

Further, the target cell is the cell containing mannose receptor.

Further, the target cell includes but is not limited to macrophage, dendritic cells and tumour cell.

The beneficial effects of the present invention are:

1) composition provided by the present invention, targeting vector, targeted drug have the target of preferable mannose receptor Tropism, can be efficiently in conjunction with the mannose receptor on target cell；

2) preparation cost of the targeting vector provided by the present invention is cheap, is readily synthesized, and has versatility, can be with A variety of targeted nano preparations are synthesized, and is conducive to purify and characterize；

3) interval insulant provided by the invention can be such that the targeting material and the basic nanometer formulation material generates Distance, therefore the targeting material can be made to be exposed to the surface of targeted nano preparation, to have efficient mannose receptor target To ability；

4) the targeted nano preparation prepared by shows good profile, and size is small, close to spherical shape, there is good serum Stability, cytotoxicity is small, is applicable to the targeting drug delivery system of drug (chemical drug and biological medicament), and transfection efficiency is significantly higher than The Lipo 3K of commercialization can be used as transfection reagent use, be applied in scientific research and business.

Detailed description of the invention

Fig. 1 is mannose-PEG₁₀₀- Chol nuclear-magnetism verification result figure.

Fig. 2 is the grain-size graph of 1 liposome of prescription.

Fig. 3 is the potential diagram of 1 liposome of prescription.

Fig. 4 is that the TEM of 1 liposome of prescription schemes.

Fig. 5 is liposome transfection result.

Fig. 6 is the expression of GFP in fluorescence microscope DC cell.

Fig. 7 is that transfection efficiency analyzes histogram.

Fig. 8 is mannose-PEG₁₀₀₀The stability analysis of-Chol.

Fig. 9 is 4 DEG C and stores lower mannose-PEG₁₀₀₀The transfection efficiency of-Chol.

Figure 10 is mannose-PEG₁₀₀₀Stability analysis of-the Chol in serum.

Figure 11 Flow cytometry mannose-PEG₁₀₀₀Toxicity of-the Chol and Lipo 3K for DC cell.

Figure 12 Flow cytometry mannose-PEG₁₀₀₀- Chol and Lipo 3K unites for the data of the toxicity of DC cell Meter analysis histogram.

Figure 13 is the research of DC2.4 cellular uptake targeted nano granule.

Figure 14 is the intake situation of mRNA targeting lipids nanocrystal composition in fluorescence microscope BMDCs cell.

Figure 15 is the data point of the intake situation of mRNA targeting lipids nanocrystal composition in fluorescence microscope BMDCs cell Analysis.

Figure 16 is that the liposome of mannose polyethylene glycol 400 cholesterol preparation carries the internal anti-swollen of the vaccine prepared after mRNA Tumor effect disquisition.

Figure 17 is that the liposome of mannose polyethylene glycol 400 cholesterol preparation carries the internal anti-swollen of the vaccine prepared after mRNA The mouse weight of tumor research monitors.

Figure 18 is that the liposome of mannose polyethylene glycol 400 cholesterol preparation carries the internal anti-swollen of the vaccine prepared after mRNA The 28th day gross tumor volume of mouse of tumor research.

Figure 19 is antitumous effect in m/MP400-LPX each group body.

Figure 20 is m/MP400-LPX immunized mice life cycle.

Figure 21 is intake of the DC2.4 cell to the different ligands chain length targeting lipids nanocrystal composition for carrying mRNA.

Figure 22 is the data point of intake of the DC2.4 cell to the different ligands chain length targeting lipids nanocrystal composition for carrying mRNA Analysis.

Figure 23 is transfection of the DC2.4 cell to different ligands chain length targeting lipids nanocrystal composition.

Figure 24 is that DC2.4 cell analyzes the transfection data of different ligands chain length targeting lipids nanocrystal composition.

Figure 25 is transfection of the BMDCs cell to different ligands chain length targeting lipids nanocrystal composition.

Figure 26 is that BMDCs cell analyzes the transfection data of different ligands chain length targeting lipids nanocrystal composition.

Figure 27 is antitumous effect in different ligands chain length target liposomes composite body.

Figure 28 is the mouse weight monitoring of antitumor research in different ligands chain length target liposomes composite body.

Figure 29 is the 28th day tumour body of mouse of antitumor research in different ligands chain length target liposomes composite body Product.

Figure 30 is antitumous effect data analysis in different ligands chain length target liposomes composite body.

Figure 31 is different ligands chain length targeting lipids nanocrystal composition immunized mice life cycle.

Specific embodiment

Illustrated embodiment is in order to which preferably the present invention will be described, but is not that the contents of the present invention are limited only to institute For embodiment.So those skilled in the art according to foregoing invention content to embodiment carry out it is nonessential improvement and Adjustment, still falls within protection scope of the present invention.

In embodiment involved mRNA according to normal conditions, such as Molecular Cloning:A Laboratory guide (third edition, J. Sa Nurse Brooker etc. write) described in condition, purchase or according to the normal condition proposed by manufacturer implement obtain.