Drugs containing galectin 9

A technology of galectin and medicine, applied in the field of medicine containing galectin 9

Inactive Publication Date: 2006-03-08
GALPHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0019] Although many physiologically active substances in the living body have been discovered so far, many of them, for example, act on normal cells as well as on tumor cells, and only a part of their activities have been clarified, so it may not be possible to simply seek to use them in medicine, etc.

Method used

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  • Drugs containing galectin 9
  • Drugs containing galectin 9
  • Drugs containing galectin 9

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0164] (1) Preparation of immunogenic antigen, (2) Immunization of animals with immunogenic antigen, (3) Preparation of myeloma cells, (4) Cell fusion of antibody-producing cells and myeloma cells, (5) Hybridoma ( fused cells) selection and monocloning, and (6) production of monoclonal antibodies

[0165] (1) Preparation of immunogenic antigen was carried out as follows. As the antigen, as described above, polypeptides isolated from natural galectin-9 polypeptides or large fragments derived therefrom (may include part of domain polypeptides, linker polypeptides, fragments, part of peptides, and synthetic polypeptides) can be used. Based on the determined amino acid sequence information of galectin 9, an appropriate oligopeptide was chemically synthesized as an antigen. Representative antigens are selected from (1) the amino acid sequence of sequence number: 1 and sequence number: 2 in the sequence listing published by WO02 / 37114 A1 or the amino acid sequence of a part of its ...

Embodiment 1

[0247] (1) Materials and methods

[0248] (a) cell culture

[0249] MOLT-4 (T cells), Jurkat (T cells), BALL-1 (B cells), THP-1 (cells from acute monocytic leukemia) and HL60 (cells from acute monocytic leukemia) were provided by the American Type Culture Collection ( ATCC) obtained. All cell lines were grown in RPMI-1640 medium (Sigma, St. Louis, USA) supplemented with 10% FCS at 5% CO 2 maintained at 37°C. In order to inhibit the activity of Gal-9, 30 mM lactose was added to the culture medium. The same concentration of sucrose was used as a control.

[0250] (b) Expression and purification of recombinant Gal-9 (rGal-9)

[0251] In accordance with well-known methods (for example, Matsushita, N. et al., J. Biol. Chem., 275: 8355 (2000); and Nishi, N. et al., Endocrinology, 141: 3194 (2000)), Make rGal-9 into (His) 6 - Galectin-9 (short form) [(His) 6 -Gal-9(S)], expressed and purified. That is, Escherichia coli BL-21 strain carrying a Gal-9 expression plasmid was gro...

Embodiment 2

[0306] (1) Cytotoxic activity of galectin-9

[0307] Cytotoxic activity was measured for the Jurkat T cell line, K-562 leukemia cells, and normal lymphocytes by flow cytometry analysis using PI. That is, after 16 hours of incubation of target cells with 1 mM galectin-9, PI was added for the last 15 minutes, and PI entered the damaged target cells. The fluorescence emitted by the PI was measured by FACS. Use cells without any other substances as a negative control, use formalin-fixed cells as 100% dead cells, and use H as a positive control 2 o 2 (hydrogen peroxide solution). As shown in Table 2, galectin-9 exhibited obvious cytotoxic activity on Jurkat cell line and K-562 cells, but did not exhibit harmful activity on normal lymphocytes.

[0308] Stimulate

[0309] (2) Cancer cell apoptosis induced by galectin-9

[0310] Apoptosis was measured by the PI method. After the cells cultured with galectin-9 were washed, they were fixed with ethanol, and the fragme...

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PUM

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Abstract

Galectin 9 exerts various functions depending on its localizations. On the other hand, galectin 9 is expected as participating in various biological functions. Thus, it has been required to clarify the detailed biological activities of galectin 9 and develop galectin 9-related techniques including development of drugs. Human galectin 9 shows a cytotoxic activity and an apoptosis-inducing activity on tumor cells but shows neither cytotoxic activity nor apoptosis-inducing activity on normal cells. Therefore, it is possible to employ galectin 9 proteins, galectin 9 agonists, galectin 9 antagonists, anti-galectin 9 binding protein antibodies, anti-galectin 9 binding sugar chain antibodies, galectin 9-producing, releasing or inducing substances, etc. as antitumor, antiallergic, immunosuppressive agents, drugs for autoimmune diseases, anti-inflammatory agents and active ingredients for adrenocortical steroid hormone alternatives.

Description

technical field [0001] The present invention relates to the use of galectin (galectin) 9, especially human galectin 9 (Gal-9) for its cytotoxic activity on malignant tumor cells, its apoptosis-inducing activity on malignant tumor cells, its Anti-tumor activity (anti-cancer activity), apoptosis-inducing activity of activated T cells, especially apoptosis-inducing activity of CD4-positive T cells, immunosuppressive activity, anti-inflammatory effect, and anti-allergic effect. The present invention relates to antitumor agents (anticancer agents), antiallergic agents, immunosuppressants, autoimmune disease agents, anti-inflammation agents and adrenal corticosteroid hormone substitute agents utilizing galectin-9 and related technologies. Background technique [0002] It has been reported that galectin-9 induces apoptosis of thymocytes in mice. On the one hand, it induces the apoptosis of T cells in rat splenocytes. On the other hand, although galectin-9 specifically induces CD8-p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K39/395A61K45/00A61K48/00A61P5/38A61P29/00A61P35/00A61P37/02A61P37/06A61P37/08A61P43/00C07K16/18
CPCA61K38/1709C07K16/18A61P29/00A61P35/00A61P37/02A61P37/06A61P37/08A61P43/00A61P5/38A61K38/17A61K39/395
Inventor 平岛光臣西望山内清明吉田尚子积正子
Owner GALPHARMA CO LTD
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