Compound-type nano-vaccine and preparation method thereof

A nano-vaccine and composite technology, applied in the field of biomedicine, can solve the problems of complex preparation, nano-carriers that cannot simultaneously load siRNA, cannot be degraded and metabolized, and achieve simple and easy preparation methods, improved anti-tumor effects, and biocompatibility good sex effect

Inactive Publication Date: 2015-05-27
SHENZHEN INST OF ADVANCED TECH
View PDF8 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there are still many deficiencies in the existing nanocarriers, such as poor stability of most nanocarriers, poor biocompatibility, inability to completely degrade and metabolize in vivo, and inability to simultaneously load multiple substances such as genes and proteins. Complicated and unable to control the particle size of nanocarriers well, the existence of these shortcomings largely limits the application of polymer nanocarriers in the preparation of nanovaccine
For example, "Nanovaccine and its preparation method" (CN 102068698A) provides a cationic liposome complex using mannosylation as a vaccine carrier to load protein antigens, but the nanocarriers in this invention cannot simultaneously load siRNA
Another example, "A preparation process for dendritic cell-targeted nanoliposome tumor seedlings" (CN101690805A), the program is to introduce α1,3Gal glycolipids into the surface of nanoliposomes to prepare DCs-targeted tumor vaccines. Nano tumor vaccine liposomes, activated DCs to enhance anti-tumor immune response, but also, this invention cannot be combined with siRNA
In addition, "A polymer gene drug carrier and its preparation method and its application in the preparation of anti-tumor drugs" (publication number CN102512683A) uses cyclodextrin hydroxypropyl cyclodextrin as the carrier skeleton to construct positively charged nanoparticles Loading siRNA that can inhibit the production of tumor vascular endothelial growth factor into tumor cells; however, this invention is only used for gene therapy and cannot be used for nano-vaccine-mediated immunotherapy

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Compound-type nano-vaccine and preparation method thereof
  • Compound-type nano-vaccine and preparation method thereof
  • Compound-type nano-vaccine and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This example is the preparation process of the polyethylene glycol monomethyl ether-polylysine-polyleucine triblock polymer carrier used in the composite nano-vaccine disclosed in the present invention.

[0038] The preparation method comprises the following steps:

[0039] (1) After evacuating the polymerization tube, fill it with nitrogen for protection, and take 1g of CH with a molecular weight of 2000 3 O-PEG-NH 2 , dissolved in 20mL N, N-dimethylformamide (DMF) and added to the polymerization tube;

[0040] (2) According to the lysine cyclic anhydride monomer and CH 3 O-PEG-NH 2 Add Lys-NCA monomer in a ratio of 10:1, and react at constant temperature for 24 hours under the protection of nitrogen;

[0041] Then, according to the leucine cyclic anhydride monomer and CH 3 O-PEG-NH 2 Add the Leu-NCA monomer with a molar ratio of 10:1, and continue the reaction at constant temperature for 24 hours under the protection of nitrogen;

[0042] After the reaction, ad...

Embodiment 2

[0047] This example is the process of using the PEG-PLys-PLeu triblock copolymer prepared in Example 1, tumor antigen (OVA), TLR3 agonist (PIC), and synthetic STAT3 siRNA to prepare a composite nano-vaccine, and the obtained polymer The structure of micelles in aqueous solution and the structure of composite nanovaccine are as follows figure 1 and figure 2 shown.

[0048] (1) Weigh 1 mg of the PEG-PLys-PLeu triblock copolymer prepared in Example 1 and dissolve it in 1 ml of ultrapure water to form a uniform and transparent polymer nanomicelle aqueous solution ( figure 1 );

[0049] (2) Mix and shake tumor antigen (OVA), TLR3 agonist (PIC), synthetic STAT3 siRNA (5′-GGAAAUUUAACAUUCUGGGCACGAA-3′, SEQ ID NO.1) and polypeptide nanomicelle aqueous solution (OVA: PIC: siRNA: NPPEG -PLL 30 -PLLeu 40 =16:4:1:79, w / w) for 1 hour, and let stand at room temperature for 30 minutes to obtain a composite nano-vaccine (see vaccine structure figure 2 ).

Embodiment 3

[0051] This example is a comparative experiment of TADC on the uptake of OVA and siRNA in the case of free state or siRNA-FAM wrapped in polypeptide nanomicelles. The experimental results are as follows image 3 shown.

[0052] 1. Experimental steps:

[0053] 1. Add PEG-PLL 30 -PLLeu 40 Polypeptide nanomicelle aqueous solution (50 μg / ml) was mixed with TLR3 agonist (PIC), tumor antigen (OVA-Alexa555) and FAM-labeled STAT3 siRNA (SEQ ID NO.1), and left to stand at room temperature for 30 minutes. Obtain a compound vaccine encapsulated by polypeptide nanomicelles (OVA: PIC: siRNA: NPPEG-PLL 30 -PLLeu 40 =5:5:1:50, w / w), namely NP / siRNA / OVA;

[0054] 2. Incubate free siRNA-FAM, free OVA-Alexa555 or NP / siRNA / OVA with TADCs (tumour-associated dendritic cells, tumor-associated dendritic cells) for 2 hours. The uptake of siRNA and OVA by TADC was detected by confocal microscopy.

[0055] 2. Experimental results:

[0056] Compared with free OVA-Alexa555 and free FAM-siRNA, pol...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
degree of polymerizationaaaaaaaaaa
degree of polymerizationaaaaaaaaaa
degree of polymerizationaaaaaaaaaa
Login to view more

Abstract

The invention discloses a compound-type nano-vaccine taking an amphiphilic three-block polymer of polyethylene glycol derivative-poly lysine-poly leucine as a nano-carrier bearing a tumor antigen, an immunological adjuvant and siRNA (si Ribonucleic Acid), and a preparation method of the compound-type nano-vaccine. According to the nano-vaccine, taking and presenting of the antigen by antigen-presenting cells can be improved by a nano-micelle bearing the antigen; siRNA is efficiently delivered to TADCs (Tumor-associated Dendritic Cells) to block immunosuppression signals of TADCs, and TADCs are induced and activated by collaboration with the immunological adjuvant, so that a tumor resisting effect of a tumor vaccine is improved; the nano-vaccine has the advantages that the adopted carrier of the nano-vaccine is good in biocompatibility and low in toxicity, and is degradable in a biological body; degradation products are nontoxic and harmless, and can be absorbed or metabolized; and the preparation method of the nano-vaccine is simple and convenient and feasible, good in stability and convenient for popularization.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a composite nano-vaccine with polymer nano-particles as a carrier and a preparation method thereof. Background technique [0002] Tumor vaccine-mediated anti-tumor immunotherapy is the fourth anti-tumor therapy after surgery, chemotherapy, and radiotherapy. It mainly uses the antigenic substance of tumor cells to stimulate the body to produce specific tumor cell immune killing, so as to achieve the purpose of eradicating tumors. Compared with traditional anti-tumor treatments, tumor vaccines have many advantages such as good safety and high specificity, and have been widely used to treat various malignant tumors such as lung cancer, colon cancer, prostate cancer, breast cancer, and melanoma. The clinical efficacy still needs to be further improved. [0003] Dendritic cells (DC), as the most powerful antigen-presenting cells in the body, are the primary core link ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/39A61K39/00A61K47/42A61P35/00
Inventor 马轶凡蔡林涛刘兰兰易虎强
Owner SHENZHEN INST OF ADVANCED TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap