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Method of screening apoptosis inducing substances

a technology of apoptosis and substances, applied in the field of screening substances, can solve the problems of scarce knowledge of the characteristics of specific antibodies recognizing surface antigens, and the hardly prepared specific antibodies, and achieve the effect of easily and highly efficient differentiation, identification and screening

Inactive Publication Date: 2003-08-21
CHUGAI PHARMA CO LTD
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  • Abstract
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Benefits of technology

[0008] This invention relates to a method of screening substances having property of causing apoptosis characterized by using cells which are expressing IAP (Integrin Associated Protein) and screening the substances. The invention relates to the screening method, wherein the cells used are myeloid cells, and relates to pharmaceutical compositions containing as the ingredient the substances obtained by the above screening method. The invention makes it possible to differentiate, identify and screen readily and highly efficiently the substances, such as antibodies and the like, that have property of causing apoptosis on myeloid cells by using cells which are expressing IAP while using specific binding reactions of the substances. The above specific substances thus obtained by the screening method of the invention can be used by virtue of their characteristics as the active ingredient of pharmaceutical compositions such as anticancer agents and medicines for myelocytic leukemia and the like which are useful in the field of remedies for myelocytic leukemia and the like.

Problems solved by technology

However, though some of splenic stromal cells have been established as a cell line (CF-1 cells), and cytological characteristics thereof have been examined, a specific antibody recognizing surface antigens thereof has been hardly prepared so far, and characteristics thereof have been scarcely known.

Method used

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[0074] Establishment of Monoclonal Antibodies

[0075] 1) Antigens and Immunization

[0076] Immunization was performed by using CF-1 cells obtained in the above Reference Example as antigens. The cells were cultured in an incubator under the condition of 5% CO.sub.2 and 37.degree. C., using an Isocove's modified Dulbecco's medium (IMDM) (Boehringer-Mannheim Co.) supplemented with 10% fetal bovine serum (FBS; Sanko Junyaku) as a medium.

[0077] The cells were treated with 1 mM EDTA / PBS, and removed from a culture flask according to pipetting. The cells were suspended into 1 mM EDTA / PBS at the cell number of about 1.times.10.sup.7 / ml, and administered to a Wistar Imamich rat (7-week-old, female; Animal Breeding Research Laboratory). One ml of cells of about 1.times.10.sup.7 / ml were injected into the abdominal cavity of the rat at the initial immunization, and 1 ml of cells of about 1.times.10.sup.7 / ml were administered additionally one month later. Further, 1 ml of cells of about 1.times.10....

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Abstract

The present invention provides a method of screening substances having property of causing apoptosis, and relates to a method of screening substances having property of causing apoptosis characterized by using cells which are expressing IAP (Integrin Associated Protein), and the relates to above screening method, wherein the cells used are myeloid cells, and relates to pharmaceutical compositions containing as the active ingredient the substances obtained by the above method, and the present invention makes it possible to differentiate, identify and screen readily and highly efficiently the substances, such as antibodies and the like, that have property of causing apoptosis on myeloid cells by using cells which are expressing IAP while using specific binding reactions of the substances, and the above specific substances thus obtained can be used by virtue of their characteristics as the active ingredient of pharmaceutical compositions such as anticancer agents and medicines for myelocytic leukemia and the like.

Description

[0001] The present invention relates to a method of screening substances having property of causing apoptosis and the like, in particular, relates to a novel screening method which makes it possible to screen readily and highly efficiently the substances, such as monoclonal antibodies and the like, that have property of causing apoptosis on myeloid cells by using cells which are expressing IAP (Integrin Associated Protein), and relates to the substances having property of causing apoptosis obtained by the above screening method, pharmaceutical compositions containing as the active ingredient the above substances, and relates to substances having property of causing apoptosis which have a specific binding activity to IAP, pharmaceutical compositions containing as the active ingredient the above substances.[0002] Granulocyte colony-stimulating factors, for example, recombinant granulocyte colony-stimulating factors (rG-CSF), have been known primarily as humoral factors to stimulate th...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00C07K14/47C07K14/705C12N15/06
CPCA61K38/00C07K14/70596C07K14/4747
Inventor FUKUSHIMA, NAOSHI
Owner CHUGAI PHARMA CO LTD
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