Anti-tumor chimeric protein, anti-tumor vaccine and anti-tumor vaccine adopting nasal mucosa drug administration mode

A chimeric protein, anti-tumor technology, applied in the field of anti-tumor vaccines, can solve the problems of inability to exert immunotherapy, weak immunogenicity, unable to recognize and present to activate toxic T lymphocytes, etc.

Inactive Publication Date: 2019-11-15
GUANGDONG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a major problem in the development of tumor vaccines is that the immunogenicity of tumor antigens is weak, and they cannot be recognized and presented by the immune system to activate toxic T lymphocytes (CTL), and cannot play the role of immunotherapy

Method used

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  • Anti-tumor chimeric protein, anti-tumor vaccine and anti-tumor vaccine adopting nasal mucosa drug administration mode
  • Anti-tumor chimeric protein, anti-tumor vaccine and anti-tumor vaccine adopting nasal mucosa drug administration mode
  • Anti-tumor chimeric protein, anti-tumor vaccine and anti-tumor vaccine adopting nasal mucosa drug administration mode

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] see figure 1 , this example provides a recombinant engineered bacterium containing the first polypeptide sequence, marked as pET22b-GM-CSF-CTA2-TAT, the construction method is as follows:

[0056] In this example, the commercial carrier pET22b was selected, and the sequence of GM-CSF-CTA2-TAT (GM-CSF, CTA2 subunit sequence of cholera toxin and TAT sequence of cell penetrating peptide sequence) was artificially synthesized and cut with NdeI and XhoI Insert it into the pET22b plasmid to obtain the recombinant plasmid pET22b-GM-CSF-CTA2-TAT for fusion expression, transform Escherichia coli BL21(DE3), and obtain the recombinant engineering bacteria containing the first polypeptide sequence.

[0057] Wherein, the amino acid sequence of GM-CSF-CTA2-TAT is:

[0058] MAPTRSPITVTRPWKHVEAIKEALNLLDDMPVTLNEEVEVVSNEFSFKKLTCVQTRLKIFEQGLRGNFTKLKGALNMTASYYQTYCPPTPETDCETQVTTYADFIDSLKTFLTDIPFECKKPSQKDPTGGASEFELGGGGSMSNTCDEKTQSLGVKFLDEYQSKVKRQIFSHRPHPDLEGTHNRIKDERLVD. The connection bet...

Embodiment 2

[0066] see figure 1 , this embodiment provides a recombinant engineered bacterium containing the second polypeptide sequence, marked as pET28a-CTB-PSMA624-632, the construction method is as follows:

[0067] In this example, the commercial vector pET28a was selected, and the amino acid sequence of CTB was recombined with pET28a through transformation to obtain the recombinant plasmid pET28a-CTB. The PSMA624-632 gene was cut with NheI and XhoI and inserted into the downstream of the CTB gene to form a recombinant plasmid for fusion expression. pET28a-CTB-PSMA624-632 was transformed into Escherichia coli BL21(DE3) to obtain a recombinant engineered bacterium containing the second polypeptide sequence.

[0068] The second polypeptide is a fusion protein, and the amino acid sequence denoted as (CTB-PSMA624-632)5 is:

[0069] MTPQNITDLCAEYHNTQIYTLNDKIFSYTESLAGKREMAIITFKNGAIFQVEVPGSQHIDSQKKAIERMKDTLRIAYLTEAKVEKLCVWNNKTPHAIAAISMANASGGTYSVSFDSLLEHHHHHH. SEQ ID No: 1 is connected to ...

Embodiment 3

[0076] This embodiment provides the expression method of the fusion protein GM-CSF-CTA2-TAT:

[0077] Cultivate the recombinant engineered bacteria containing the first polypeptide sequence of Example 1 at 37°C, with ampicillin and 0.1mmol / L isopropyl-β-D-thiopyran galactobacter at a final concentration of 50 μg / mL Glycoside (IPTG) induces the expression of the fusion protein to obtain the first polypeptide (GM-CSF-CTA2-TAT).

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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to an anti-tumor chimeric protein, an anti-tumor vaccine and an anti-tumor vaccine adopting a nasal mucosadrug administration mode. The provided anti-tumor chimeric protein comprises first polypeptide comprising a lymphokine sequence used for enhancing an immunoreaction and a CTA2 subunit sequence of a cholera toxin, and second polypeptide comprising a CTB subunit sequence of the cholera toxin and a tumor specific antigen epitope peptide sequence; a CTA2 subunit of the cholera toxin of the first polypeptide and a CTB subunit of the cholera toxin of the second polypeptide are subjected to mutual chimerism to obtain the anti-tumor chimeric protein. The provided anti-tumor chimeric protein, anti-tumor vaccine and anti-tumor vaccine adopting the nasal mucosa drug administration mode are used for overcoming the technical defect that in the prior art, an existing tumor vaccine is poor in immunogenicity.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an anti-tumor chimeric protein, an anti-tumor vaccine and an anti-tumor vaccine for nasal mucosa administration. This application claims the rights and interests of the Chinese patent (patent application number 201811119727.6) submitted on September 25, 2018, and the entire content of the above application is hereby incorporated herein by reference. Background technique [0002] Tumor vaccines are a hot spot in oncology research in recent years. Through the administration of tumor-specific antigens or antigenic polypeptides, they can activate, restore or strengthen the body's anti-tumor immune response, and then kill and eliminate tumor cells expressing tumor-specific antigens. However, a major problem in the development of tumor vaccines is that the immunogenicity of tumor antigens is weak, and they cannot be recognized and presented by the immune system to activ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00A61K39/00A61K39/39A61P35/00
CPCA61K39/00A61K2039/543A61K2039/55522A61K2039/55544A61P35/00C07K14/28C07K14/4748C07K14/535C07K2319/10C07K2319/40C07K2319/55
Inventor 王华倩林丹敏何华锋孙家杰龙威郑希卢宇靖杜志云赵肃清张焜
Owner GUANGDONG UNIV OF TECH
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