Small molecule, indirubin-3-oxime, for prevention and treatment of bone disease

Abstract

The present invention provides a composition comprising an indirubin derivative for stimulating longitudinal bone growth. Because the composition for stimulating longitudinal bone growth according to the present invention is shown to be effective in longitudinal bone growth, it may be used not only as a composition for stimulating longitudinal bone growth, but also as a pharmaceutical composition for treating or preventing short stature, microplasia, dwarfism, or precocious puberty.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to Korean Application No. 10-2013-0081464 filed on Jul. 11, 2013, which is incorporated herein by reference.TECHNICAL FIELD

[0002] The present invention relates to a small molecule, indirubin-3-oxime, for prevention and treatment of bone disease.BACKGROUND ART

[0003] The increased awareness of the importance of normal growth, as well as an increased aesthetic interest, in infants and adolescents has led to an increased public interest in height growth. In the past, height growth was thought to depend on heredity. However, it is said that the proportion to which heredity is responsible for actual height growth is only 23%, and also, depending on the care during the period of growth, sufficient height growth can be achieved. Therefore, a great deal of money is being invested, estimated to be anywhere between 4-100 million dollars, to achieve height growth within the time period in which the growth plate is not c...

Examples

example 1

Cell Culture and Reagents

[0026]Rat chondrosarcoma (RCS) cells were cultured in a Dulbecco's modified Eagle's medium (DMEM; Gibco, Grand Island, N.Y.), supplemented with 10% fetal bovine serum (FBS) and penicillin / streptomycin.

example 2

Western Blot Analysis

[0027]For Western blot analysis, cells were treated with indirubin-3′-oxime for 24 hours, cultured and reached to 70% confluency and then were harvested. After the harvested samples underwent cytolysis, the proteins were quantified, which was followed by electrophoresis using 10% or 12% SDA-PAGE gel. Afterwards, immunoblot was conducted using antibodies.

example 3

Immunocytochemistry

[0028]2.5×104 cells were adhered to a cover glass placed in a 12-well plate. Once securely attached, cells were treated with indirubin-3′-oxime for 24 hours, and then were rinsed with a phosphoric acid buffer solution. After adding 4% paraformaldehyde, cells were fixed at room temperature for 20 min. Then, proteins expressed in the cells were observed through cell membrane permeation, blocking, and immunofluorescent staining processes.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to Korean Application No. 10-2013-0081464 filed on Jul. 11, 2013, which is incorporated herein by reference.TECHNICAL FIELD

[0002] The present invention relates to a small molecule, indirubin-3-oxime, for prevention and treatment of bone disease.BACKGROUND ART

[0003] The increased awareness of the importance of normal growth, as well as an increased aesthetic interest, in infants and adolescents has led to an increased public interest in height growth. In the past, height growth was thought to depend on heredity. However, it is said that the proportion to which heredity is responsible for actual height growth is only 23%, and also, depending on the care during the period of growth, sufficient height growth can be achieved. Therefore, a great deal of money is being invested, estimated to be anywhere between 4-100 million dollars, to achieve height growth within the time period in which the growth plate is not c...

Examples

example 1

Cell Culture and Reagents

[0026]Rat chondrosarcoma (RCS) cells were cultured in a Dulbecco's modified Eagle's medium (DMEM; Gibco, Grand Island, N.Y.), supplemented with 10% fetal bovine serum (FBS) and penicillin / streptomycin.

example 2

Western Blot Analysis

[0027]For Western blot analysis, cells were treated with indirubin-3′-oxime for 24 hours, cultured and reached to 70% confluency and then were harvested. After the harvested samples underwent cytolysis, the proteins were quantified, which was followed by electrophoresis using 10% or 12% SDA-PAGE gel. Afterwards, immunoblot was conducted using antibodies.

example 3

Immunocytochemistry

[0028]2.5×104 cells were adhered to a cover glass placed in a 12-well plate. Once securely attached, cells were treated with indirubin-3′-oxime for 24 hours, and then were rinsed with a phosphoric acid buffer solution. After adding 4% paraformaldehyde, cells were fixed at room temperature for 20 min. Then, proteins expressed in the cells were observed through cell membrane permeation, blocking, and immunofluorescent staining processes.

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