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Micelle polypeptide vaccine with poly(ethylene glycol)-phosphatidylethanolamine as carrier

A pegylated phospholipid and polypeptide vaccine technology, which is applied in the field of medicine and biology, can solve the problems of few studies on protein or polypeptide entrapment and no reports, and achieve the effect of improving the cure rate and survival rate

Active Publication Date: 2015-12-30
PRESONE WUHAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few studies on the encapsulation of proteins or polypeptides in PEGylated phospholipid micelles, and there are no reports at home and abroad on the research on tumor therapeutic vaccines developed using PEGylated phospholipid micelles as carrier systems

Method used

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  • Micelle polypeptide vaccine with poly(ethylene glycol)-phosphatidylethanolamine as carrier
  • Micelle polypeptide vaccine with poly(ethylene glycol)-phosphatidylethanolamine as carrier
  • Micelle polypeptide vaccine with poly(ethylene glycol)-phosphatidylethanolamine as carrier

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1, the preparation of micelle E7 vaccine

[0066] The E7 antigen polypeptide is a tumor-associated antigen derived from human papillomavirus type 16 (HPV16), and a palmitic acid molecule is connected to the N-terminal, that is, the E7-20 polypeptide; E7 43-62 Sequence: Palmitate-GQAEPDRAHYNIVTFCCKCD;

[0067] Stock solution preparation:

[0068] (1) Add 1mL of anhydrous methanol to each tube of 1mg E7-20 that has been dispensed to make a stock solution of E7-20 with a concentration of 1mg / mL;

[0069] (2) Dissolving the MPLA powder in a 1:2 mixed solution of methanol and chloroform to prepare a MPLA stock solution with a concentration of 1 mg / mL;

[0070] (3) Weigh 30 mg of PEG-PE powder and add 3 mL of chloroform to dissolve to prepare 10 mg / mL PEG-PE stock solution.

[0071] 1. Preparation of micellar vaccine solution, prepare 10mL micellar vaccine according to the prescription in Table 1:

[0072] Step (1) Take 5mL of PEG-PE stock solution in a rotary ...

Embodiment 2

[0081] Example 2, Determination of Encapsulation Efficiency of Fatty Acidated E7 / OT-1 Antigen Micelles

[0082] Synthesize fatty acidated E7 antigenic polypeptide and OT-1 antigenic polypeptide (palmitic acid-EQLESIINFEKLTEWKD) with rhodamine fluorescence modification, the free fattyated antigenic polypeptide cannot dissolve in water and thus form a suspension, and the fattyated antigenic polypeptide is PEG -After the PE micelles are loaded, a stable solution is formed, so that the free antigenic polypeptide and the antigenic polypeptide loaded by the micelles can be separated, and then calculated according to the change of the content of the antigenic polypeptide loaded by the micelles in the solution Encapsulation rate of antigen polypeptide, encapsulation rate = packaged antigen amount / total antigen amount x100%.

[0083] According to the ratios in Table 2 and Table 3, micelles containing different proportions of polypeptides were prepared, and the preparation process was t...

Embodiment 3

[0088] Embodiment 3, the physical properties characterization of micellar polypeptide vaccine

[0089] 1. Morphological observation of micellar E7 vaccine by transmission electron microscope

[0090] Dilute the micellar E7 vaccine solution prepared in Example 1 with deionized water to dilute the PEG-PE concentration to 0.1 mg / mL, take 10 μL of the sample solution, and add it dropwise to the carbon-coated surface treated with glow discharge hydrophilization After 30 seconds of adsorption on the membrane copper grid, absorb the sample solution with filter paper, add 10 μL of uranyl acetate (concentration: 2% (w / v)) dropwise and stain for 30 seconds, absorb the staining solution with filter paper, and use a transmission electron microscope (Tectai20 ) to observe the micelle morphology after negative staining. The result is as figure 1 As shown, the micellar vaccine was observed by transmission electron microscope to be uniform and spherical nanoparticles.

[0091] 2. Particle...

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Abstract

The invention relates to a micelle polypeptide vaccine with poly(ethylene glycol)-phosphatidylethanolamine as a carrier. The vaccine can prevent or treat tumors or can be used as a combined preparation of antitumor active preparations. The micelle polypeptide vaccine is formed by self-assembling of poly(ethylene glycol)-phosphatidylethanolamine (PEG-PE) and antigenic polypeptide. The PEG-PE is a compound formed by binding polyethylene glycol (hydrophilic block) to nitrogenous base on lipid molecule (hydrophobic block) through a covalent bond. Particle size of the micelle vaccine is 10-100 nm. The antigenic polypeptide loaded in the micelle polypeptide vaccine is a polypeptide vaccine with length of 5-100 amino acids. According to needs, the micelle polypeptide vaccine also contains an immunologic adjuvant for regulating body immunity.

Description

technical field [0001] The invention belongs to the field of medicine and biology, and in particular relates to a micellar polypeptide vaccine with pegylated phospholipid as a carrier, its preparation method and application. Background technique [0002] Tumor therapeutic vaccines designed with nanoscale materials as carriers are new ideas for tumor treatment. Existing studies have shown that the use of nanoscale material carriers to carry protein antigens can improve the immunogenicity of antigens. This phenomenon may be related to the relatively large specific surface area and complex structure of nanoscale materials; It can protect to a certain extent and prevent protein antigens from being quickly degraded by proteases and intranet systems in the blood, which is conducive to the function of protein antigens; in addition, nanoscale material carriers carrying protein antigens are easily captured by antigen presenting cells (antigenpresenting cells). , APCs), especially de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/385A61K39/00A61K39/12A61K9/107A61K47/34A61P35/00
Inventor 梁伟秦焱周畅
Owner PRESONE WUHAN BIOTECHNOLOGY CO LTD
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