Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Hormones – e.g. – prolactin – thymosin – growth factors – etc.
Patent
1996-03-05
1997-07-22
Green, Lora M.
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Hormones, e.g., prolactin, thymosin, growth factors, etc.
530350, 930120, C07K 14475, C07K 1451
Patent
active
056504950
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a novel protein, more particularly, to an animal brain derived protein having an avidity with a heparin-binding growth factor.
TECHNICAL BACKGROUND
Recently, many growth factors have been isolated from various animal tissues and animal cells, and their characters have been elucidated. Among these growth factors is known a group of proteins called heparin-binding growth factors (HBGFs) having a high affinity with heparin. Among the most widely known HBGFs, there are acidic fibroblast growth factors (aFGF) and basic fibroblast growth factors (bFGF) found in the 1930's. Among those found recently, OSF-1 (Japanese Patent Kokai No. 117398/1992) or HBGF-8 (Japanese Patent Kokai No. 159299/1992) can be cited. Although OSF-1 and HBGF-8 were found independently, they are the identical substance.
These proteins are known to have a differentiation-enhancing action to neuronal cells and proliferation-enhancing action to osteoblasts, and they are considered to be possibly involved in the diseases accompanied with decrease of nerve function and decrease of osteogenetic potency. Therefore, if factors that connect with those proteins and control the actions of those proteins are found, they are expected to be applied to therapeutics of neuronal function diseases and osteoblast potency diseases. However, reported until present is confined only to a part of such factors which connect with heparin-binding growth factors and control their action, such as FGF receptors for FGF, and a satisfactory result has not yet been obtained for the application of these factors to a therapeutic drug. Yet, there is still no report on any binding proteins for recently found OSF-1 (HBGF-8).
Then, the present inventors have attempted to find out a protein having an avidity to this factor, by focusing on OSF-1 (HBGF-8) among heparin-binding growth factors and finally completed this invention.
DISCLOSURE OF THE INVENTION
An object of the present invention is to provide a novel protein which is useful as a therapeutic drug for neuro-functional and osteogenetic diseases by finding out the proteins connecting with OSF-1, one of heparin-binding growth factors having a differentiation-enhancing action to neuronal cells and proliferation-enhancing action to osteoblasts, and controlling these actions.
The present inventors paid their attention to the fact that OSF-1 exists abundantly in animal brains, and have attempted to isolate proteins having an avidity with OSF-1 from brains, based on a hypothesis that the proteins which connect with OSF-1 should exist also in brains. Namely, the present inventors thought it possible to isolate OSF-1 and the proteins having an avidity with it from brains in the state as they are binding, by isolating OSF-1 from brains under a mild condition. The present inventors thought it also possible to isolate, thereafter, an OSF-1 binding protein by releasing the bondage of OSF-1 and desired protein. These have been proved by experiments, which led to the completion of the present invention.
The brains from juvenile rats of 14 days after birth was used because it was anticipated that such brains especially had larger amount of various growth factors with high activity. The brains were purified until OSF-1-binding proteins were made homogeneous by using centrifugation, ion-exchange column chromatography and reverse phase column chromatography. Thereafter, sequencing of amino terminal sequence of the protein was carried out to find out that the protein provided in the present invention is an entirely novel protein, and it was named HBP-200.
The protein provided in the present invention is a novel protein isolated from animal brains, having the following physical properties and amino terminal sequence; condition) the Sequence Listing
Furthermore, the proteins provided in the present invention can be isolated also from bone tissues, osteoblasts, osteoblast strains, fibroblasts and fibroblast strains in which OSF-1 (HBGF-8) is known to exist.
BRIEF EXPLANATION OF THE DRAWINGS
FI
REFERENCES:
Gieffers et al., Eur. J. Cell Biol. 62 (2): 352-361 (1993).
Katoh et al., Cancer Res. 53 (5): 1136-1141 (1993).
Kuo et al., J. Biol. Chem. 265 (31): 18749-18752 (1990).
Experimental Medicine 10 (1): 25-31 (1992).
Raulo et al. J. Biol. Chem. 267 (1992) 11408-11416.
Doi Kazuyuki
Kimura Michio
Green Lora M.
Hayes Robert C.
Hoechst Japan Limited
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