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Nanofiber antitumor vaccine formed by self-assembly peptide folding and method

A technology of nanofibers and self-assembled peptides, applied in the fields of molecular biology and immunology, can solve problems such as limited immunogenicity, inability to produce a strong enough anti-tumor immune response, lack of curative effect of vaccines, etc., achieve significant healing effect, suppress immunity Effect of suppressor cell MDSC, significant anti-tumor immune effect

Active Publication Date: 2019-08-20
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, current vaccines usually lack significant efficacy in clinical trials, and one of the possible reasons is that they cannot generate a sufficiently strong antitumor immune response. The immunogenicity of E6 and E7 protein or peptide-based subunit vaccines is usually limited, Therefore, exploring effective antigen delivery vehicles may be an important approach to enhance the clinical efficacy of tumor vaccines

Method used

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  • Nanofiber antitumor vaccine formed by self-assembly peptide folding and method
  • Nanofiber antitumor vaccine formed by self-assembly peptide folding and method
  • Nanofiber antitumor vaccine formed by self-assembly peptide folding and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation and morphological identification of nanofiber anti-tumor vaccine

[0043] HPV16 E7 44-62 The peptide was chemically modified to the N-terminus of the self-assembling peptide Q11 through the flexible arm Ser-Gly-Ser-Gly, and the E7-Q11 peptide was purified by reverse-phase HPLC, lyophilized, and stored at -20°C. Dissolve lyophilized E7-Q11 in sterile water to 2mM, and place it at 4°C overnight, then assemble E7-Q11 into nanofibers with 1×PBS (pH7.4) solution, the assembly condition is 1×PBS (pH7.4 ) solution to dilute the E7-Q11 aqueous solution to 0.5 mM, and stand at room temperature 20° C. for 4.5 hours to obtain an anti-HPV-related tumor vaccine assembled into nanofibers.

[0044] The assembled E7-Q11 nanofibers were observed by a transmission electron microscope (TEM) for the morphology of the assembled E7-Q11 fibers.

[0045] Among them, Q11 peptide (amino acid sequence: Ac-Gln-Gln-Lys-Phe-Gln-Phe-Gln-Phe-Glu-Gln-Gln-Am, such as SED IQ NO.4...

Embodiment 2

[0051] Example 2: Preventive immunization strategy and mouse transplantation tumor model establishment

[0052] Female C57BL mice, 6-8 weeks old, in the preventive immunization experiment, the mice were first immunized 3 times with an interval of 2 weeks, and the mice were divided into two groups, with 5 mice in each group, which were the immune Q11 carrier group ( Using Q11 peptide) and E7-Q11 vaccine group (using nanofiber E7-Q11), the immunological dose is 12.5nmol, each 200ul, two weeks after the third immunization, subcutaneously inject a volume of 50ul in the right abdomen of the mouse, 1 ×10 5 TC-1 cells (cell concentration 2×10 6 cells / ml), and the volume ratio of cells and Basement MembraneMatrix is ​​1:1 ( image 3 ). After tumor inoculation, tumor size was measured every 3-4 days ( Figure 4 ). Twelve weeks after the first tumor inoculation, the tumor was re-inoculated with 1 × 10 subcutaneously in the left abdomen. 5 TC-1 cells (cell concentration 2×10 6 cel...

Embodiment 3

[0053] Example 3: Establishment of transplanted tumor models in mice and therapeutic immunity to 2-3mm tumors

[0054] The mice were divided into 3 groups, and 1 × 10 5 TC-1 cells (cell concentration 2×10 6 cells / ml), immunization procedures such as Figure 5 , when the tumor grows to 2-3mm, the vaccine is immunized, and the mice are divided into three groups, namely the Q11 vector group, the E7-Q11 vaccine group, and the E7-Q11 non-assembly group (the E7-Q11 non-assembly group is E7-Q11 Immediately immunize mice with lyophilized peptide dissolved in sterile water (without 1×PBS assembly), the immunological dose is 12.5nmol, 100ul each time, immunized 3 times in total, with one week interval, and the size of the tumor is measured every 3-4 days ( Figure 6 , Figure 7 ), 34 days after the third immunization, the spleen of the mice was taken for immunological detection, using ELISPOT to detect antigen-specific lymphocytes secreting IFN-γ, and using E7 49-57 Peptides were u...

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Abstract

The invention relates to a nanofiber antitumor vaccine formed by self-assembly peptide folding and a method and belongs to the technical field of molecular biology and immunology. HPV16 E744-62 peptide is linked to an N fragment of self-assembly peptide Q11 through Ser-Gly-Ser-Gly to be assembled in a 1*PBS solution, nanofiber E7-Q11 is formed by self-assembly, and the nanoscale fiber form of thenanofiber E7-Q11 is indentified by a transmission electron microscope. After immunizing mice, the nanofiber E7-Q11 can induce organisms to produce strong HPV16 E7 specific cellular immune response andsignificantly inhibit growth of tumor.

Description

technical field [0001] The invention belongs to the technical fields of molecular biology and immunology, and in particular relates to a nanofiber anti-tumor vaccine formed by folding self-assembled peptides and a construction method of the vaccine. Background technique [0002] Cervical cancer is the second most common malignant tumor that threatens women's health around the world. Human papillomavirus (HPV) is a key pathogenic factor of cervical cancer, and HPV is detected in almost all cervical cancer cases. Among them, HPV16 is a high-risk type of HPV, which is related to 50% of cervical cancer. In addition, HPV is also associated with a variety of tumor diseases, such as penile cancer, vaginal cancer, anal cancer, oropharyngeal cancer, head and neck cancer, etc. Expression of HPV oncoproteins E6 and E7 leads to the transformation of infected cells and the proliferation of malignant cells by promoting the cell cycle of infected cells, inhibiting apoptosis and increasing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K47/64A61P35/00A61P31/20
CPCA61K39/12A61K47/64A61P35/00A61P31/20C12N2710/20034Y02A50/30
Inventor 马雁冰李思瑾张启书谢航航白红妹黄惟巍孙文佳
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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