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A dual-target tumor vaccine and its preparation method and application

A tumor vaccine and dual-target technology, applied in the field of biomedicine, can solve the problems of weak immunogenicity and small polypeptide molecules, and achieve the effects of promoting apoptosis, improving effectiveness and safety, and promoting cascade system.

Active Publication Date: 2021-08-27
AFFILIATED YONGCHUAN HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In view of this, one of the objectives of the present invention is to provide a dual-target tumor vaccine, which not only retains the advantages of safety and easy preparation and purification of polypeptide vaccines, but also overcomes the small polypeptide molecules, weak immunogenicity, and difficulty in being easily absorbed in vivo. Insufficient uptake of antigen-presenting cells (APCs), etc., can stimulate effective specific cytotoxic T lymphocyte (CTL) responses in vivo, and at the same time play the role of inhibiting tumor angiogenesis and inducing tumor cell apoptosis

Method used

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  • A dual-target tumor vaccine and its preparation method and application
  • A dual-target tumor vaccine and its preparation method and application
  • A dual-target tumor vaccine and its preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Preparation of Example 1 GPR124 Gene Recombination Eukaryotic Expression Plasmid pcDNA3.5 / GPR124

[0042] (1) Cloning of the full-length coding gene of GPR124

[0043] According to the GPR124 gene sequence with the GenBank accession number NM_054044.2, PCR primers were designed and synthesized to amplify the full-length coding gene of GPR124; PCR was performed using the human placenta cDNA library as a template. Denaturation at ℃ for 30 seconds, annealing at 56℃ for 30 seconds, extension at 72℃ for 30 seconds, a total of 30 cycles, and finally extension at 72℃ for 5 minutes. Ligated with the vector pGEM-T, the ligated product was transformed into Escherichia coli JM109 competent cells, and the medium containing Amp, IPTG, X-GAL was used for blue and white spot screening, the white spot was picked and cultured, the plasmid was extracted, and the gene sequence was determined by Shanghai Shenggong Company , named the positive clone plasmid with correct sequence as pGEM-T / ...

Embodiment 2

[0048] Preparation of the fusion peptide of embodiment 2 membrane-penetrating peptide and AVPI peptide

[0049] The fusion peptide is formed by directly linking the carboxyl terminus of the penetrating peptide and the amino terminus of the AVPI peptide, and its amino acid sequence is shown in SEQID No.1.

[0050] The fusion peptide was solid-phase synthesized on an AB-431A peptide synthesizer, using a standard fluorenylmethoxycarbonyl (Fmoc) protocol, with 0.25 mmol of p-hydroxymethylphenoxymethyl polystyrene (HMP) resin as the starting resin, According to the amino acid sequence of SEQ ID No.1, the peptide chain is extended from the carboxyl terminal to the amino terminal one by one. After the synthesis of the peptide chain is completed, the resin containing the peptide chain is transferred to a cutting solution (0.25 mL of ethylenediamine tartrate, trifluoroacetic acid 9.5mL and 0.25mL of deionized water), stir the reaction at room temperature to crack the peptide chain from...

Embodiment 3

[0053] The preparation of embodiment 3 tumor vaccines

[0054] At room temperature under the condition of vortexing, add 1 volume part of fusion peptide with a concentration of 1 mg / ml dropwise to 1 volume part of an aqueous solution containing pcDNA3.5 / GPR124 at a concentration of 1 mg / ml and PBS with a concentration of 1 mg / ml Peptide solution, after the dropwise addition, continued to swirl for 60 minutes, and then stood still for 60 minutes to obtain the tumor vaccine.

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Abstract

The invention belongs to the field of biomedicine, and in particular relates to a dual-target tumor vaccine and its preparation method and application. The tumor vaccine of the present invention is composed of tumor vascular endothelial marker GPR124 recombinant eukaryotic expression plasmid and fusion peptide through electrostatic interaction, and GPR124 recombinant eukaryotic expression plasmid is composed of GPR124 gene and eukaryotic expression vector. The dual-target tumor vaccine of the present invention has the following advantages: 1) The tumor vaccine has dual targets, which can simultaneously inhibit tumor angiogenesis and induce tumor cell apoptosis; 2) The vaccine is safe, easy to prepare and purify, has strong immunogenicity, and is easily infected APC uptake; 3) Greatly improve the effectiveness and safety of tumor treatment, and have good application prospects in the field of tumor treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a dual-target tumor vaccine and its preparation method and application. Background technique [0002] Tumor is a disease with high morbidity and high mortality that seriously threatens human health. Angiogenesis is a necessary condition for the growth of primary and metastatic tumors. As early as the 1930s and 1940s, some scholars found that the absolute number of angiogenesis in tumor tissue increased sharply. Once there are new blood vessels in the tumor, the tumor cells proliferate rapidly, the tumor grows rapidly, and it is easy to infiltrate and metastasize. [0003] After surgery, radiotherapy and chemotherapy, biological therapy has become an important means of comprehensive treatment of tumors. At present, tumor biological treatment strategies include targeted therapy of tumor-related genes or tumor suppressor genes, immunotherapy of tumor-associated antigens, etc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/00A61K38/17A61K47/64A61K47/69A61K48/00A61P35/00
CPCA61K38/177A61K39/0011A61K48/005A61K2039/523A61K2039/53A61K2039/57A61K47/64A61K47/6901A61P35/00
Inventor 杨曌
Owner AFFILIATED YONGCHUAN HOSPITAL OF CHONGQING MEDICAL UNIV
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