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Human interferon alpha-2b recombinant bacillus calmette-guerin, construction method and identification method thereof

A technology of recombinant BCG and interferon alpha, applied in recombinant DNA technology, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of high incidence of toxic side effects, high incidence of side effects, high cost, etc. The effect of improving the anti-bladder cancer effect and reducing the application dose and side effects

Inactive Publication Date: 2009-03-11
丁国庆
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Problems solved by technology

However, clinically, there are still about 30% of patients with unsatisfactory BCG treatment and prevention effects, especially for invasive bladder cancer.
In addition, BCG intravesical infusion has a high incidence of toxic side effects
In recent years, many studies have used low-dose BCG combined with intravesical infusion of human interferon α-2b (IFNα-2b) to improve anticancer efficacy and reduce side effects, but the long-term efficacy of combined application is not good, and exogenous added IFNα-2b has the problems of large application dose, high incidence of side effects, short duration of action, and high cost

Method used

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  • Human interferon alpha-2b recombinant bacillus calmette-guerin, construction method and identification method thereof
  • Human interferon alpha-2b recombinant bacillus calmette-guerin, construction method and identification method thereof
  • Human interferon alpha-2b recombinant bacillus calmette-guerin, construction method and identification method thereof

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Embodiment Construction

[0030] 1. Principle of the present invention:

[0031] Studies believe that BCG's anti-bladder cancer effect is due to the body's immune response after BCG perfusion, and cytokines such as interleukin-2 (IL-2), gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF) -α), etc. play an important role in the process of anti-cancer. Increasing the expression of these cytokines can improve the immune function and anti-cancer efficacy of BCG. IFN α-2b is currently one of the most widely used cytokines in clinical practice, and is widely used in anti-tumor and anti-viral therapy. It can not only directly act on tumor cells, inhibit tumor cell proliferation and induce differentiation, but also studies have shown that IFNα-2b combined with BCG perfusion in the bladder can increase the expression of local IL-2, IFN-γ, TNF-α and other cytokines. Thereby improving the immune activity and anti-cancer ability of BCG. However, exogenous application of IFN α-2b has the problems of large dose,...

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Abstract

The invention relates to a recombinant BCG vaccine rBCG-IFN alpha-2b for secreting human IFN alpha-2b and a construction method and an identification method thereof, wherein a BCG vaccine Ag85B signal peptide fragment which has the function of secretion and genes of human IFN alpha-2b are cloned to pMV261 by the genetic engineering technology, and a BCG vaccine shuttle expression vector pMV261-Ag85B-IFN alpha-2b is obtained; and the vector is induced into BCG by the electrotransformation technology, and the recombinant BCG vaccine rBCG-IFN alpha-2b is established; and the human IFN alpha-2b can be highly efficiently secreted in virtue of the secretion function of the pMV261-Ag85B-IFN alpha-2b on BCG replication and signal peptide. The recombinant BCG vaccine rBCG-IFN alpha-2b obtained not only keeps the immunogenicity of the prior BCG but also can continuously secrete the cell factor-the IFN alpha-2b, thereby improving the immunocompetence of the BCG; the IFN alpha-2b can be directly acted on tumor cells to inhibit proliferation and induced differentiation of the tumor cells, has good antitumor action, and can reduce the application dosage and reduce the toxic and side effect caused by the BCG; and the IFN alpha-2b solves the problems of large application dosage, high incidence rate of side effects, short response time and expensive cost caused by exogenous IFN alpha-2b.

Description

Technical field: [0001] The invention relates to an anti-bladder cancer drug in clinical medicine, in particular to a recombinant BCG vaccine of human interferon alpha-2b, and a construction method and identification method thereof. Background technique: [0002] Transitional epithelial carcinoma of the bladder is the most common malignant tumor of the urinary system in my country, and it is easy to recur and progress after surgery. Therefore, mitomycin, doxorubicin, thiotepa, Bacille Calmette-Guerin (BCG) and other drugs should be regularly used for bladder perfusion to prevent tumor recurrence. Among them, BCG is currently considered the most effective drug. However, clinically, about 30% of patients still have unsatisfactory BCG treatment and prevention effects, especially for invasive bladder cancer. In addition, BCG bladder perfusion has a higher incidence of side effects. In recent years, many studies have applied low-dose BCG combined with human interferon α-2b (IFNα-2b) b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/21C12N15/74A61K39/04A61P35/00C12Q1/68G01N33/68
Inventor 丁国庆沈周俊
Owner 丁国庆
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