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Oral vaccine delivery system and application thereof

A drug delivery system and oral vaccine technology, applied in the field of new pharmaceutical preparations, can solve the problems of small size of nanoparticles, loss of nanoparticles, large surface activity, etc., and achieve good biocompatibility, simple process, and improved immunogenicity. Effect

Inactive Publication Date: 2016-12-14
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, the size of the nanoparticles is small, there are a large number of unsaturated bonds on the surface, and the surface activity is very high. Coupled with van der Waals force and electrostatic force, the nanoparticles dispersed in the liquid medium are prone to coagulation and agglomeration, forming secondary bonds. Particles, making the particle size larger, and eventually losing the unique functions of nanoparticles when used

Method used

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  • Oral vaccine delivery system and application thereof
  • Oral vaccine delivery system and application thereof
  • Oral vaccine delivery system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Scheme screening of acid-resistant HP55 / PLGA-CCF NPs

[0060] Option One:

[0061] (1) Weigh 40mg of PLGA and 20mg of HP55 in the prescribed amount, dissolve in a mixed solution of 1mL of acetone and 1mL of dichloromethane, and prepare the drug-loaded organic phase for use;

[0062] (2) Weigh 60 mg of poloxamer 188 and dissolve in 6 mL of ultrapure water to make a hydration medium solution as W 1 / O / W 2 The external aqueous phase of the system. Weigh 30mg of poloxamer 188 and dissolve it in 30mL of ultrapure water to make a hydration medium solution, which is used as the continuous phase for nanoparticle dispersion;

[0063] (3) Dissolve the dual epitope-dual adjuvant Helicobacter pylori vaccine CTB-UE-CF (CCF) in refolding buffer (50mmol / L NaH 2 PO4, 10 mmol / L TisHCl, 2 mmol / L GSH, 0.4 mmol / L GSSH, adjust the pH value to 8.0 with HCl or NaOH), the concentration is about 4.2mg / mL, as the internal water phase of the double emulsion (W 1 ). Add 500 μL of ...

Embodiment 2

[0073] Example 2: Dynamic research on immune level of HP55 / PLGA-CCF NPs vaccine.

[0074] (1) Grouping scheme of SPF BALB / c mice: (A) NC group: Mice were orally administered a suspension of 500 μL PBS and aluminum hydroxide adjuvant. (B) HP55 / PLGA-group: Mice were orally administered acid-resistant HP55 / PLGA nanoparticles without antigen encapsulation. (C) HP55 / PLGA-CCF group: Mice were orally administered acid-resistant HP55 / PLGA nanoparticles encapsulated with 100 μg CCF. (D) PLGA-CCF group: Mice were orally administered ordinary PLGA nanoparticles encapsulated with 100 μg CCF. (E) Alum-CCF group: Mice were orally administered a suspension of 100 μg CCF and aluminum hydroxide adjuvant.

[0075] (2) Mice immunization and challenge program: according to Image 6 , orally immunized 4 times by intragastric administration, with an interval of 7 days each time, and challenged the immunized mice two weeks later: intragastric administration of Hp suspension (1×10 9 CFU / mL), 0.3...

Embodiment 3

[0081] Example 3: HP55 / PLGA-CCF NPs vaccine induces a high level of systemic immune response.

[0082] ELISA detection of specific antibodies IgG, IgM and IgA in antiserum before and after challenge:

[0083] ELISA reagents: (A) Coating solution: Beijing Solaibao Technology Co., Ltd. (B) Washing solution: weigh 0.2g KH 2 PO 4 , 2.9gNa 2 HPO 4 •12H 2 O, 8.0g NaCl, 0.2g KCl, 0.5mL Tween-20, add ddH 2 O was adjusted to 1000mL (PBST). (C) Blocking solution: Weigh 3.0 g of BSA and dissolve in 100 mL of washing buffer, filter and sterilize and store at 4°C. (D) Sample diluent: Weigh 1.0 g of BSA and dissolve in 100 mL of washing buffer, filter and sterilize and store at 4°C. (E) Substrate solution: soluble one-component TMB substrate solution. (F) Stop solution: measure 178.3mL of distilled water, add 21.7mL of concentrated sulfuric acid (1M H 2 SO 4 ).

[0084] Detection protocol: Dilute the natural urease antigen to 10 μg / mL with coating solution, add 100 μL to each we...

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Abstract

The invention discloses an oral vaccine delivery system. W1 of W1 / O / W2 type nanoparticles is a vaccine medicine phase, an organic phase O is selected from PLGA and HP55, an external water phase W2 and a continuous water phase are selected from poloxamer 188, and a vaccine medicine is a double epitope-double adjuvant helicobacter pylori vaccine. The method adopts a polymer material having good biological safety and good compatibility to prepare the nanoparticles, a preparation process and a formula are optimized, the oral vaccine delivery system is non-toxic, free of any side effects, easy to manufacture and low in cost. In the field of biological medicines, the nanoparticles can be used for oral taking of a helicobacter pylori subunit vaccine, accordingly helicobacter pylori infection related diseases are prevented and treated, and huge economic benefits and social benefits are brought.

Description

technical field [0001] The invention belongs to the field of new formulations of pharmaceutical preparations, and relates to the preparation of HP55 / PLGA nanoparticles with resistance to gastric acid degradation and its application in an oral vaccine delivery system. Background technique [0002] Helicobacter pylori ( Helicobacter pylori , Hp) is a Gram-negative, microaerophilic bacterium that primarily colonizes the stomach. The normal population infection rate is over 50%. It produces a large amount of urease, which decomposes urea in the stomach to generate ammonia and neutralizes gastric acid, so that it can colonize the gastric mucosa and epithelial layer, causing chronic gastritis, peptic ulcer, mucosa-associated tissue lymphoma, and even gastric cancer. At present, the treatment of Hp infection is mainly based on the "triple combination" therapy, namely the combination therapy of antibiotics, proton pump inhibitors and bismuth. However, this therapy has disadvantag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02A61K9/51A61K47/34A61K47/38A61P31/04
CPCA61K39/0208A61K9/5146A61K9/5161A61K2039/542A61K2039/55505
Inventor 奚涛邢莹莹谭周林刘巍刘海
Owner CHINA PHARM UNIV
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