Method for preparing dendritic cells with capacity to induce cancer-specific immune response, and pharmaceutical composition and kit incorporating dendritic cells for preventing or treating cancer or inhibiting metastasis

A technology of dendritic cells and immune response, applied in the direction of drug combination, blood/immune system cells, biochemical equipment and methods, etc., can solve the problems such as the disappearance of imiquimod anti-proliferation effect

Inactive Publication Date: 2012-05-02
JW PHARMA CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the function of OGFr was blocked by siRNA technology, the antiproliferative effect of imiquimod also disappeared (9)

Method used

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  • Method for preparing dendritic cells with capacity to induce cancer-specific immune response, and pharmaceutical composition and kit incorporating dendritic cells for preventing or treating cancer or inhibiting metastasis
  • Method for preparing dendritic cells with capacity to induce cancer-specific immune response, and pharmaceutical composition and kit incorporating dendritic cells for preventing or treating cancer or inhibiting metastasis
  • Method for preparing dendritic cells with capacity to induce cancer-specific immune response, and pharmaceutical composition and kit incorporating dendritic cells for preventing or treating cancer or inhibiting metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-1

[0109] Example 1-1: Preparation of recombinant antigen protein

[0110] Using HepG2 and SK-BR-3, which are human liver cancer cell lines and breast cancer cell lines, to synthesize cDNA and clone liver cancer specific antigens AFP (alpha-fetoprotein), MAGE 1 (melanoma associated family A1), and GPC 3 (glypican3).

[0111] If it is AFP, select the 20-346 amino acid sequence in the full-spectrum amino acid sequence of SEQ ID NO: 1 for the expression site; if it is GPC 3, the 33-358 amino acid sequence in the full-spectrum amino acid sequence of SEQ ID NO: 2 sequence, In the case of MAGE 1, select the full spectrum amino acid sequence in the sequence of SEQ ID NO:3. Furthermore, an antigen protein in which peptide YGRRARRRRRR was fused to the above-mentioned selected protein was used.

[0112] After cloning the antigen cDNA synthesized in the YGRRARRRRRR peptide-fusion expression vector, it was transformed into Escherichia coli expression strain BL21-DE3, and a transformant that...

Embodiment 1-2

[0113] Example 1-2: Preparation of tumor cell lines expressing human liver cancer antigens

[0114] As a tumor cell line for liver cancer mouse subjects, the same type of liver cancer cell line MH234 cell line as the C3H mouse series transformed with a eukaryotic expression vector (pcDNA3.1, Invitrogen) was used to stably express Human AFP, MAGE 1, GPC 3.

Embodiment 1-3

[0115] Examples 1-3: Isolation of bone marrow cells (precursor cells of immature dendritic cells) for dendritic cell preparation

[0116] Mouse (C3H line) bone marrow cells (bone marrow cells) come from femur and tibia. In order to obtain the femur and tibia, the mice were first sacrificed by cervical dislocation, and the surgical site was sterilized with 70% ethanol. The femur and tibia were separated, sterilized by soaking in 70% ethanol for about 30 seconds, washed with phosphate buffered saline (PBS), and soaked in RPMI-1640 medium. Use sterilized surgical scissors to cut off both ends of the femur and tibia, inject RPMI-1640 medium into a 1cc syringe, insert an injection needle into the cut site and squeeze it, then fill it in a 50ml tube to obtain bone marrow cells. The obtained cells are centrifuged, and after the bone marrow cells are washed once, the red blood cells in the bone marrow cells are removed. After removing erythrocytes, centrifugation was performed twice...

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Abstract

The present invention relates to a method for preparing mature dendritic cells, having a significantly improved capacity for inducing a cancer-specific immune response; a pharmaceutical composition for preventing or treating a cancer or inhibiting cancer metastasis, containing the mature dendritic cells prepared by using the above method; and a pharmaceutical kit incorporating the mature dendritic cells and an imiquimod prepared by the above method. Accordingly, it is possible to prepare safe mature dendritic cells which significantly improves the induction capacity of the cancer-specific immune response and lowers toxicity. The mature dendritic cells are effective in preventing and treating cancer, and inhibiting metastasis. In parallel with administering the cancer-specific mature dendritic cells, the imiquimod used for preparing the pharmaceutical kit improves cancer immunotherapy and prevents or treats a cancer and inhibits cancer metastasis.

Description

technical field [0001] The present invention relates to a method for preparing mature dendritic cells that can improve the inducibility of cancer-specific immune responses, and to a method for preventing or treating cancer or inhibiting cancer metastasis, including mature dendritic cells prepared based on the method. A pharmaceutical composition, and a pharmaceutical kit comprising mature dendritic cells and immune adjuvant prepared based on the method. Background technique [0002] Most of the cancer treatment methods known so far are surgery, anticancer drugs, and radiation therapy. However, these treatments are difficult to cure and have many side effects. In particular, most metastatic cancers or recurrent cancers cannot be operated on, and chemotherapy agents are rejected, so the treatment is limited. [0003] Recently, cancer therapy using immune cells such as dendritic cells (Dendritic cells) is actively being developed. Dendritic cells capture and activate externa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0784C12N5/02C07K7/06A61K35/12A61P35/00A61K35/15A61K39/00
CPCC12N5/0639A61K35/15A61K2039/5154A61P35/00
Inventor 裴容洙闵美璟成英林张瑱我尹仙玉
Owner JW PHARMA CORP
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