Anti-tumor agent using interleukin-23 gene

A technology of interleukin and anti-tumor agent, which is applied in anti-tumor drugs, gene therapy, medical preparations containing active ingredients, etc., can solve the problems of high clearance rate, short half-life, difficult to maintain concentration of cytokines, etc. The effect of tumor metastasis

Inactive Publication Date: 2005-11-30
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high clearance rate of the body, the half-life of general cytokines is very short, and it is difficult to maintain a certain concentration of cytokines in the blood, and excessive administration of cytokines has toxic effects. Therefore, direct administration of recombinant cytokines has certain limitations.

Method used

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  • Anti-tumor agent using interleukin-23 gene
  • Anti-tumor agent using interleukin-23 gene
  • Anti-tumor agent using interleukin-23 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Comparison of IL-23 and IL-12 Secreted by Activated Dendritic Cells

[0038] After the mouse bone marrow T cells, B cells and macrophages were removed by antibody method, the non-adherent cells were cultured in the medium supplemented with GM-CSF and IL-4 (20ng / ml, PeproTech Company) for 10 days to obtain the tree shaped cells. Cell Sorter was used to analyze the surface markers of the obtained cells, and cell activation markers such as type II major histocompatibility antigen and CD86 were weakly positive, so they were determined to be immature dendritic cells.

[0039] These cells (6×10 5 cells / well) and lung cancer cell A11 cells (2×10 5 cells / well) after co-cultivation, CD86 and other cell activation markers increased, confirming that the cells were activated. At this time, p40 was detected from the culture supernatant by ELISA (BioSourse kit), but as a control, p40 was not detected in the supernatant after dendritic cells were co-cultured with FasLigand gene-int...

Embodiment 2

[0042] Establishment of IL-23 Gene Transferred Cells

[0043] Total RNA was extracted from BALB / c mouse subcutaneously transplanted tumor cells, and the full-length sequence cDNA of p19 (sequence 3) and p40 (sequence 4) was obtained by RT-PCR method, and inserted into the retroviral vector LXSN (Miller AD, Rosman GJ, BioTechniques 7; 980-990, 1989) multiple cloning site (EcoRI / Bam HI), the two cDNAs are connected by internal ribosome entry sites (internal ribosome entry sites, IRES) ( Duke GM et al., J Virol 66:1602-1609, 1992). Therefore, the retrovirus can transcribe p19 (located at the 5' end of the IRES) and p40 (located at the 3' end of the IRES) under the action of the promoter located at the long terminal repeat (LTR), and the transcripts can be translated independently.

[0044] The above-mentioned retroviral vector was transfected into packaging cells ψ2 cells using Lipofectin reagent (Invitrogen Company), and then G418 (400 μg / ml, Invitrogen Company) was added to th...

Embodiment 3

[0047] Anti-tumor Effect of IL-23 Gene Transferred into Cells

[0048] Inoculation of two kinds of Colon 26 cells (Colon26 / IL-23#1, #9) with IL-23 gene introduced into syngeneic BALB / c mice resulted in delayed tumor growth, and the tumor completely regressed after a period of time. As a control, after inoculation of parental cells with empty vector and no gene introduced, the tumors grew rapidly, eventually leading to the death of the mice. Other clones also had similar antitumor effects. Colon 26 cells (1×10 6 1) within 20 days, all the mice died. After inoculating the same amount of Colon 26 / IL-23#9 or #12, most of the mice survived for more than 40 days (χ 2 =3.5, df=1, P=0.037; log-rank test, P=0.046).

[0049] Colon 26 / IL-23#1 and Colon 26 / IL-23#9 were mixed at a ratio of 1:9 (the total cell number was 1×10 6 1) and inoculated into the same line of mice, the speed of tumor formation and growth was significantly slower than that of Colon 26 parent strain alone (P<0.05...

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Abstract

The invention belongs to the biology technical field, relating an antineoplastic agent taking IL-23 gene or a cell introduced to the IL-23 gene as effective ingredient. The invention comprises the antineoplastic agent taking the following substances as the effective ingredient: a virus representing the IL-23 gene or non-virus carrier, a tumor cell introduced the IL-23 gene losing fission ability and an immune cell introduced the IL-23 gene. The antineolastic agent of the invention can be used independently or be combined with the treatment methods of operation, actinotheraphy and medical treatment for improving the cell immune response of organism, thereby achieving the aim of decreasing tumor volume, preventing transfer and preventing the recurrence of the tumor.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an antitumor agent with IL-23 gene or cells introduced with IL-23 gene as active ingredients. Specifically, it includes anti-tumor agents with the following substances as active ingredients: viral or non-viral vectors expressing IL-23 gene, tumor cells that have been introduced with IL-23 gene and have lost their ability to divide, and immune cells that have been introduced with IL-23 gene. The antitumor agent of the present invention can be used alone or combined with surgery, radiotherapy, chemotherapy and other treatment methods to improve the body's cellular immune response, thereby reducing tumor volume, preventing metastasis and tumor recurrence. Background technique [0002] Existing tumor treatment methods (such as radiation therapy, chemotherapy, surgical resection, etc.) are not conducive to the prognosis of patients because they have great toxic and side effects and easily c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09A61K38/00A61K48/00A61P35/00
Inventor 田川雅敏王彦青
Owner FUDAN UNIV
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