Active immune regulation particle, and preparation method and application thereof

An active immunization and microparticle technology, applied in chemical instruments and methods, pharmaceutical formulations, vertebrate antigen components, etc., can solve the problems of poor anti-tumor effect of vaccines, break immune tolerance, promote amplification, and improve immune response Effect

Active Publication Date: 2020-05-15
优锐生物医药科技(深圳)有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the shortcomings of the prior art described above, the purpose of the present invention is to provide an active immune regulati...

Method used

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  • Active immune regulation particle, and preparation method and application thereof
  • Active immune regulation particle, and preparation method and application thereof
  • Active immune regulation particle, and preparation method and application thereof

Examples

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preparation example Construction

[0062] The preparation method of the active immune regulation particles of the present invention at least includes the following steps:

[0063] 1) preparing SYNZIP2-CpG-microcarrier particles;

[0064] 2) Connect SYNZIP1 to T cell promoter, antibody Fc fragment and antigen respectively to obtain connector I, connector II and connector III;

[0065] 3) Mix linker I, linker II and linker III with SYNZIP2-CpG-microcarrier particles to obtain the active immune regulation particles.

[0066] The preparation method of described SYNZIP2-CpG-microcarrier particle comprises the steps:

[0067] 1) Mixing SYNZIP2 with an activator for activation; mixing the activated SYNZIP2 with a cross-linking agent for a cross-linking activation reaction to obtain a mixture I;

[0068] 2) activating by mixing the nucleic acid molecule CpG with an activator; mixing the activated nucleic acid molecule CpG with a cross-linking agent to obtain a mixture II;

[0069] 3) Dissolving the microcarrier in a...

Embodiment 1

[0083] Example 1 Construction process of active immune regulation microparticle assembly

[0084] 1. SYNZIP-1-FcOP protein

[0085] SYNZIP-1-FcOP protein is obtained by gene synthesis. Specifically, get:

[0086]

[0087]

[0088] SYNZIP-1 -linker-FcOP amino acid sequence (30KD)

[0089] MEFGLSWLFLVAILKGVQCNLVAQLENEVASLENENETLKKKNLHKKDLIAYLEKEIANLRKKIEE (GGGGS) 3 PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNATYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIAATISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFALYCSLTVNDKSLHFSG

[0090] (SEQ ID NO: 12)

[0091] 2. Cloning of IL-15-SYNZIP1 gene

[0092] Restriction sites EcoR I and Not I were designed at both ends of IL-15-SYNZIP1, and Suzhou Jinweizhi Biotechnology Co., Ltd. was commissioned to synthesize them by whole-gene synthesis to obtain the PUC19-IL-15-SYNZIP1 cloning plasmid.

[0093] The obtained PUC19-IL-15-SYNZIP1 cloning plasmid and PTT5 plasmid were respectively digested wit...

Embodiment 2

[0107] Example 2 Preparation of Active Immunomodulatory Particles

[0108] 1. Preparation of PLGA nanospheres

[0109] 20mg SYNZIP2 (sulfo-SMCC) cross-linking activation reaction: first, SYNZIP2 activation, dissolve SYNZIP2 with 10ml PBS, add TCEP, open disulfide bond; dialyze to remove TCEP, and get activated SYNZIP2. Then add sulfo-SMCC, dialyze to remove excess sulfo-SMCC after cross-linking, and obtain 20 mg of SYNZIP2-sulfo-SMCC solid powder after freeze-drying.

[0110] (1) Cross-linking reaction of nucleic acid molecule CpG (CpG ODN 2006 and ODN M362) 10 mg with sulfo-SMCC: First, CpG is activated, CpG is dissolved with WFI, TCEP is added to open the disulfide bond; then TCEP is removed by desalting to obtain activated CpG .

[0111] Then add sulfo-SMCC, after cross-linking, alcohol sedimentation and centrifugation to obtain solid CpG-sulfo-SMCC.

[0112] The sequence of CpG ODN 2006 is (TCGTCGTTTTGTCGTTTTGTCGTT) (SEQ ID NO: 19)

[0113] The sequence of CpG-SH ODN M...

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Abstract

The present invention provides an active immune regulation particle. The active immune regulation particle comprises a microcarrier, the microcarrier is loaded with at least SYNZIP1 polypeptide, SYNZIP2 polypeptide, nucleic acid molecule CpG, a T cell promoter, an antibody Fc segment and an antigen, an amino acid sequence of the SYNZIP1 polypeptide is shown in SEQ ID NO:1 and an amino acid sequence of the SYNZIP2 polypeptide is shown in SEQ ID NO:2. The active immune regulation particle can overcome immune suppression environment of tumor microenvironment, breaks immune tolerance, improves immune response of the entire immune system to tumor antigens, quickly and efficiently activate immune responses of tumor-specific B cells and T cells to kill tumors, and can also promote proliferation of the immune cells through stimulation by cytokines. Through administration by subcutaneous injection, the active immune regulation particle can be passively targeted into tumor tissues through capillaries and quickly endocytosed by the immune cells and tumor cells.

Description

technical field [0001] The invention relates to the field of vaccine preparations, in particular to an active immune regulation microparticle and its preparation method and application. Background technique [0002] Therapeutic vaccine (Therapeutic Vaccine) is a new concept of tumor immunotherapy established and developed in recent years, which refers to a vaccine that can break the immune tolerance in chronically infected patients and rebuild or enhance the immune response. The basic principle of tumor therapeutic vaccines is to improve the ability of the immune system to recognize and kill cancer cells containing such antigenic peptides by targeting tumor-associated or tumor-specific antigenic peptides. The specific steps include the following four steps: 1) After the exogenous antigen enters the body, it is taken up by antigen-presenting cells (APC) in the form of endocytosis, and then hydrolyzed in the cell into molecules that can bind to MHC-I or MHC-II molecules. Anti...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K9/16A61K47/64A61P35/00C07K17/08C07K14/00A61K38/20A61K39/00
CPCA61K39/39558A61K39/0011A61K38/2086A61K9/1658A61K9/0019A61P35/00C07K17/08C07K14/00A61K2300/00Y02A50/30
Inventor 倪健梁辉崔大祥
Owner 优锐生物医药科技(深圳)有限公司
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