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Novel polypeptide for resisting tumors caused by EB (Epstein-Barr) viruses, and application and preparation method thereof

一种EB病毒、肿瘤的技术，应用在病毒肽、抗肿瘤药、抗病毒剂等方向，能够解决机体异常免疫应答、不能用药、多肽类药物过敏反应等问题，达到降低可能性的效果

Inactive Publication Date: 2011-06-22
AMERICAN PHEROMYCIN BIOTECHNOLOGY CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in the above-mentioned anti-tumor polypeptides, due to the presence of amino acid residues that are more likely to cause hypersensitivity reactions at the sleeping end of colistin polypeptides, it is possible that drugs containing Escherichia coli polypeptides are more likely to cause abnormal immune responses in the body; some research reports, Many cancer patients are abnormal because their metabolic mechanism is disturbed by cancer cells, and they are prone to allergic reactions to peptide drugs, which leads to inability to use drugs. cancer drug

Method used

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  • Novel polypeptide for resisting tumors caused by EB (Epstein-Barr) viruses, and application and preparation method thereof
  • Novel polypeptide for resisting tumors caused by EB (Epstein-Barr) viruses, and application and preparation method thereof
  • Novel polypeptide for resisting tumors caused by EB (Epstein-Barr) viruses, and application and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Construction of a recombinant plasmid containing the gene encoding mutant colicin Ia

[0053] The original plasmid is pSELECT loaded with colicin polypeptide Ia and immunity protein gene TM -1 plasmid (8.3kb) (purchased from Promega Company), through double-stranded oligonucleotide point mutation technology (QuickChange TM Kit, Strategene Company) respectively operably link the gene encoding the mutated amino acid such as the oligonucleotide primer sequence shown in SEQ ID NO 1 to 10 with the gene of wild-type colicin Ia to obtain the sequence shown in SEQ ID NO. The gene encoding the colicin Ia allosteric polypeptide shown in 23 is obtained at the same time as a mutant plasmid; then the gene SEQ ID NO.26 or SEQ ID NO.28 encoding the mimetic antibody is inserted into the gene for the colicin Ia allosteric polypeptide in the mutant plasmid After the I626 codon of the new Epstein-Barr virus, two recombinant plasmids pCHCEB11 (such as figure 1 indicated), pCH...

Embodiment 2

[0127] Example 2. Observation of the immune effect of the novel anti-tumor polypeptide prepared by recombinant plasmids pCHCEB11 and pCHCEB22

[0128]Take the novel anti-tumor polypeptide 1 and novel anti-tumor polypeptide 2 prepared by the recombinant plasmids pCHCEB11 and pCHCEB22 prepared in Example 1, and each of the anti-tumor polypeptide 1 and anti-tumor polypeptide 2 (ZL200410081446.8) in the inventor's previous patent Partial immunization of mice: After mixing the above proteins with adjuvant, the basic amount and supplementary amount are intraperitoneally injected 50 μg (0.5ml) per mouse once. Two weeks apart, 5 injections. ELISA indirect method detects mouse serum titer, novel anti-tumor polypeptide 1 and 2 immunized mouse serum titer that the present invention makes is 10 -3 to 10 -4 And anti-tumor polypeptide 1, the serum titer of anti-tumor polypeptide 2 immunized mice is 10 -4 to 10 -5 .

[0129] It can be seen that the possibility of the novel anti-tumor po...

Embodiment 3

[0130] Example 3. Hypoallergenic Effect Experiment of Colistin Ia Allosteric Polypeptide Constituting Novel Anti-tumor Polypeptide

[0131] Take the mutant plasmid of the colistin Ia allosteric polypeptide in Example 1 (the amino acid residues G11A, H22G, A26G, V31L and H40D have been mutated in the peptide chain of its aqueous pore structure domain), and it is operable at its amino or carboxyl terminal Two kinds of antibacterial polypeptides were prepared by linking Staphylococcus aureus pheromone AgrD1 (YSTCDFIM). The polypeptide prepared by linking AgrD1 at the carboxyl terminus of allosteric colistin Ia is named anti-aureus polypeptide 1, and the polypeptide prepared by linking AgrD1 at the amino terminus of allosteric colistin Ia is named anti-Pseudomonas aeruginosa polypeptide 1 . Take the plasmid of wild-type colicin Ia, and link the pheromone AgrD1 of Staphylococcus aureus to its amino terminal to prepare the anti-Pseudomonas aeruginosa polypeptide 2.

[0132] Experi...

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Abstract

The invention relates to a novel polypeptide for resisting tumors caused by EB (Epstein-Barr) viruses, and application and a preparation method thereof, belonging to the fields of antineoplastic medicaments. The novel polypeptide is composed of a colicine allosteric polypeptide capable of forming ion channels, and an antibody polypeptide for resisting EB viruses or a mimic antibody polypeptide ofan anti-EB virus antibody, wherein the colicine allosteric polypeptide capable of forming ion channels is formed by mutating amino acid residues G11A, H22G, A26G, V31L and H40D with wild type colicine E1, Ia, Ib, A, B and N or a peptide chain of aqueous pore canal domain thereof; and the amino acid sequence of the anti-EB virus antibody is identical to that of a monoclonal antibody secreted by ATCC HB-168 hybridoma. The invention provides an improved medicament, which has the advantages of strong lethality, high safety, high specificity and low sensitization possibility, for treating tumors caused by EB viruses, and a preparation method thereof.

Description

technical field [0001] The invention relates to the field of antitumor drugs, in particular to a novel anti-Epstein-Barr virus-induced tumor polypeptide and its application and preparation method. Background technique [0002] In the field of antibiotic research, people have been committed to developing new antibiotics by imitating the way bacteria of the same strain kill each other. There are many bacterial toxins in nature that directly form ion channels on the bacterial membrane to kill bacteria. The type specimen is a bacterial toxin secreted by Escherichia coli - colicin. Among them, after colicin Ia was discovered by Jacob in 1952, after several generations of efforts, Qiu et al (Major transmemebrane movements engaged with colicin Ia channel gating. J. Gen. Physiology, 107: 313-328 (1996)) in 1996 The transmembrane three-dimensional structure of the ion channel formed by colistin Ia on the artificial lipid bimolecular membrane when it is opened and closed was finally ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/63C07K14/245C12N15/31A61K39/108A61K47/48A61P35/00A61P31/20
CPCC07K16/085C07K19/00A61K2039/505C12N2710/16222C07K14/245C07K2319/30C07K14/05C07K14/005A61P31/20A61P35/00C12N15/62C12N15/11
Inventor 丘小庆
Owner AMERICAN PHEROMYCIN BIOTECHNOLOGY CO
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