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Sufficiently diverse amphiphilic MHC II binding polypeptide, immunocarrier microsphere and preparation method and application thereof

A technology that combines peptides and amphiphilic properties. It is used in medical preparations without active ingredients, medical preparations containing active ingredients, peptides, etc. It can solve problems such as protein leakage, difficulty in realizing clinical therapeutic value, etc., and achieve specificity. Good results

Active Publication Date: 2019-05-17
SHANGHAI WEIQIU BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0022] This creates a new problem: due to the clearance of B cell clones with high affinity for endogenous proteins, the abundance of B cells that bind to exogenous carrier protein epitopes with high affinity is higher than that of B cells that bind to endogenous protein epitopes. The abundance of cells is several orders of magnitude higher, so there is a problem of immune competition for protein epitopes in the molecule of cross-linked proteins (or fusion proteins)
However, the corresponding antibodies of endogenous proteins are inhibited, and only low-titer antibodies can be produced, which is difficult to achieve clinical therapeutic value.
[0023] 2. Specificity problem: Only high-titer antibodies can play a therapeutic role
The problem with this technology is that the protein in the microspheres is easy to leak out of the microspheres, resulting in high-titer antibodies against the carrier protein
On the other hand, the application only produces antibody immune effects

Method used

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  • Sufficiently diverse amphiphilic MHC II binding polypeptide, immunocarrier microsphere and preparation method and application thereof
  • Sufficiently diverse amphiphilic MHC II binding polypeptide, immunocarrier microsphere and preparation method and application thereof
  • Sufficiently diverse amphiphilic MHC II binding polypeptide, immunocarrier microsphere and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Design and Preparation of MHCII-binding Polypeptide Self-Assembly Amphipathic Polypeptide Used in Mouse Animal Experiments

[0070] 1. MHCII-binding peptides for Balb / C mice, tetanus exotoxin fragments were selected according to the literature:

[0071] TT830-844 QYIKANSKFIGITELD

[0072] 2. For the MHCII-binding peptides of C57 / BL6 mice, egg ovalbumin fragments were selected according to the literature:

[0073] OVA 324-338 ISQAVHAAHAEINEA

[0074] 3. For the construction of immune carrier microspheres, the design and synthesis of amphipathic polypeptides containing the above two MHCII-binding polypeptides can be synthesized by conventional methods:

[0075] (1) Pal-AAAAGGG-C-QYIKANSKFIGITELD-RDKQKK-mal (Pal: palmitoyl, mal: maleimide).

[0076] (2) Pal-AAAAGGGG-C-ISQAVHAAHAEINEA-QKDRDK-mal

Embodiment 2

[0077] The preparation of microspheres in embodiment 2 organic solvent / water solvent mixed solvent

[0078] 1. The principle of peptide self-assembly into balls:

[0079] Such as figure 1 As shown, the amphiphilic polypeptide belongs to the polymer surfactant, and at the water / oil two-phase interface, the hydrophobic end is immersed in the oil phase, and the hydrophilic end is free in the water phase. If the oil phase is in the shape of microspheres, the amphiphilic polypeptide can be used as the mold of the oil phase microspheres, and beta-parallel aggregated and distributed across the interface of the microspheres. Due to the existence of two kinds of β-sheet peptides, when the solvent in the oil phase evaporates under stirring conditions, the amphiphilic polypeptide is insoluble in water and becomes a stable self-assembled microsphere in the water phase.

[0080] 2. Preparation operation

[0081] (1) 10 mg of the synthesized amphipathic MHCII-binding polypeptide is disso...

Embodiment 3

[0087] Example 3 Preparation of solid lipids as template polypeptide self-assembled microspheres

[0088] 1. The principle of ball formation: first dissolve palmitic acid in an organic solvent, emulsify into microspheres as a template. The amphiphilic polypeptide solution is poured into it and mixed with it, and under stirring condition, the hydrophobic end of the amphiphilic polypeptide is buried in the organic phase containing palmitic acid to be anchored, and the hydrophilic end is in the water phase. After the organic solvent volatilizes to form solid particles, it forms microspheres with stable dispersion. Particle size can be controlled by ultrasonic or homogenizer. The proviso is that the solvent in which the amphipathic polypeptide is dissolved is immiscible with the organic phase in which the palmitic acid is dissolved.

[0089] The invention uses acetonitrile / water to dissolve the amphipathic polypeptide, and palmitic acid is dissolved by petroleum ether or cyclohe...

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Abstract

The invention relates to a sufficiently diverse amphiphilic MHC II binding polypeptide, an immunocarrier microsphere and a preparation method and application thereof. The sequence structure of the amphipathic MHC II binding polypeptide is fatty acyl-beta-pleated sheet peptide-C-MHC II binding polypeptide-hydrophilic beta-pleated sheet peptide-maleimide, and an amphipathic MHC II binding polypeptide forms the immunocarrier microsphere by self-assembly. Coupling groups are located on the surface of the microsphere. The microsphere is particularly suitable for the preparation of antibody vaccinesfor the treatment of benign diseases and the preparation of antibody vaccines for the treatment of tumors. The antigen can be coupled to the coupling groups on the surface of the immunocarrier microsphere. The antibody vaccines can also be used in combination with FcR-CART cells or NK cells.

Description

[technical field] [0001] The invention relates to the field of immune carrier microspheres, in particular to sufficiently diverse amphipathic MHC II binding polypeptides, immune carrier microspheres and their preparation methods and applications. [Background technique] [0002] 1. Therapeutic vaccines [0003] Vaccination is one of the effective means for humans to resist and control infectious diseases. Vaccines in the traditional sense are mainly preventive vaccines, which are applied to susceptible animals and people in healthy groups. The vaccine components are generally microbial protective antigens, which can activate specific immune responses in healthy bodies, produce specific antibodies and Cytotoxic T lymphocytes (CTLs) to acquire immune protection against the pathogen. [0004] Therapeutic vaccine (Therapeutic Vaccine) is a new concept of immunotherapy established and developed in recent years, which refers to a vaccine that can break the immune tolerance in chr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K47/64A61K47/69A61P35/00C07K7/08A61K35/17
CPCY02A50/30
Inventor 谈立松
Owner SHANGHAI WEIQIU BIOTECH
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