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Therapeutic interleukin-4 vaccine capable of treating human or animal tumor diseases

A therapeutic vaccine and interleukin technology, applied in gene therapy, anti-tumor drugs, allergic diseases, etc., can solve the problems of short half-life of antibodies, high cost, easy to cause allergic reactions, etc.

Active Publication Date: 2015-05-20
济南翰宝生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the short half-life of the antibody (the half-life of IgG is about 21-28 days), repeated injections are required, which not only increases the cost, but also easily causes allergic reactions

Method used

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  • Therapeutic interleukin-4 vaccine capable of treating human or animal tumor diseases
  • Therapeutic interleukin-4 vaccine capable of treating human or animal tumor diseases
  • Therapeutic interleukin-4 vaccine capable of treating human or animal tumor diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Construction and vaccine preparation of mouse IL-4 gene recombinant vaccine

[0016] According to the whole gene coding sequence of mouse IL-4, a pair of suitable primers were designed to amplify IL-4 cDNA sequence by RT-PCR method. Then, after the DNA sequence was determined to prove that the amplified cDNA sequence was correct, it was cloned into the appropriate part of the pET-32 fusion expression vector, and the recombinant plasmid and the pET-32 fusion expression vector blank control plasmid were further transferred into Escherichia coli strains for proper expression were used for large-scale fermentation to extract and purify the IL-4 fusion protein and the thioredoxin expressed by the vector control. After mixing the purified IL-4 fusion protein or the thioredoxin expressed by the carrier control with oil adjuvant or aluminum gel adjuvant in an appropriate proportion [according to the content of the purified IL-4 fusion protein, for oil adjuvant vaccin...

Embodiment 2

[0019] Example 2: Safety test and allergic reaction test of mouse IL-4 gene recombinant vaccine

[0020] According to the method of Example 1, the purified IL-4 fusion protein or the thioredoxin expressed by the carrier control was properly diluted with normal saline and then mixed with oil adjuvant or aluminum gel adjuvant at a volume ratio of 1:1 to prepare IL-4 fusion protein. -4 fusion protein vaccine or vehicle control thioredoxin vaccine. The prepared IL-4 fusion protein vaccine was divided into three different dose groups of low (0.1ml, about 1 dose), medium (0.25ml, about 2.5 doses), high (0.5ml, about 5 doses) to immunize BALB respectively / c mice, pET-32 vector vaccine and normal saline control group were set up separately, 5 mice in each group, injected subcutaneously at multiple points. The second and third immunization injections were performed at intervals of 3 weeks. After the vaccine was injected, the mice were observed for adverse reactions, allergies, and d...

Embodiment 3

[0021] Embodiment 3: The test of enhancing anti-tumor activity of mouse IL-4 gene recombinant vaccine

[0022] According to the method of Example 1, the purified IL-4 fusion protein or the thioredoxin expressed by the carrier control was properly diluted with physiological saline and then mixed with aluminum gel adjuvant at a ratio of 1:1 to prepare the experimental or control vaccine. It is required that the amount of IL-4 fusion protein or carrier control thioredoxin in each vaccine is about 15-20 μg; the volume of each vaccine is about 0.1 ml. The experimental mice were inoculated intraperitoneally (i.p.) with the IL-4 gene recombinant aluminum gel vaccine prepared in Example 1 3 weeks and 1 week before the tumor cell attack, respectively, and a pET-32 carrier aluminum gel vaccine and normal saline immunized control group were also set up . Tumor growth was checked every 3 days after tumor inoculation, and the experiment ended at 60 days. The results showed that the IL-4 ...

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Abstract

The invention discloses a therapeutic interleukin-4 vaccine capable of treating human or animal tumor diseases. The therapeutic interleukin-4 vaccine is an any-form protein vaccine or conjugated protein vaccine prepared by taking a natural or artificially-synthesized intact protein or protein fragment of interleukin-4 as an antigen or an any-form gene vaccine or fused gene vaccine prepared by taking an intact gene or gene fragment of interleukin-4 as an antigen gene or main antigen gene. According to the invention, a host is subjected to active immunotherapy by using an IL-4 vaccine, generally, the effective time of the first immunity can be up to about 2-3 months, the effective treatment time of the second immunity can be up to about 6 months, and the rehabilitation aim can be achieved through carrying out immunotherapy for 1-3 times. Compared with a method for treating human or animal tumor diseases by directly applying an IL-4 antibody, the therapeutic interleukin-4 vaccine has the characteristics of low application frequency, small dosage and the like, so that not only is the treatment cost greatly reduced, but also the possibility for generating anaphylactic reaction is greatly reduced.

Description

technical field [0001] This case is a divisional application for the Chinese patent application "Interleukin-4 Therapeutic Vaccine for Treating Human or Animal Immune-Related Diseases" (application number: 2011101749184). Background technique [0002] So far, people still lack more effective treatment methods for chronic infectious diseases caused by intracellular parasitic pathogens (viruses, bacteria, parasites, etc.), and the preventive effect of vaccines is not satisfactory. Theoretically, such chronic infectious diseases require the host to produce a strong type (Th1) anti-infection immune response to eliminate pathogens, but how to induce the body to produce a strong Th1 anti-infection immune response is still a difficult problem. According to the applicant's long-term exploration and research, most of the clinical stages of such chronic infectious diseases show a strong second-type (Th2) anti-infection immune response, that is, the Th2 anti-infection immune response i...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/00A61P37/00A61P31/00A61P33/00A61P37/08A61P35/00
Inventor 刘永庆
Owner 济南翰宝生物技术有限公司
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