Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing heterocyclic carbon compound having only o – n – s,...
Patent
1992-10-09
1994-05-17
Fox, David T.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing heterocyclic carbon compound having only o, n, s,...
435 41, 4352404, 43524046, 43524048, 549510, C12P 1702, C12P 100, C12N 500, C12N 502
Patent
active
053127404
DESCRIPTION:
BRIEF SUMMARY
DESCRIPTION
1. Technical Field
The present invention relates to cell culture of plants belonging to the genus Taxus. More specifically, the present invention relates to a process for producing taxol by cell culture of plants belonging to the genus Taxus.
Taxol is an alkaloid of diterpene, which exhibits a carcinostatic activity and inhibits cytokinesis.
2. Background Art
Taxol is a compound promising as an antitumor agent. In particular, for ovarian cancer, taxol has been subjected to the Phase III clinical trial sponsored by the National Cancer Institute (NCI) of National Institute of Health in the U.S.A. In the known process, taxol is produced by an extraction from the bark of the wild Pacific Yew tree, Taxus brevifolia NUTT. However, since the Pacific Yew plants grow very slowly, the regrowth of the bark after removal thereof is not expected, and further, the content of taxol therein, and the production efficiency thereof, are very low. For example, as much as 10 tons of the bark are required for producing as little as 1 kg of taxol. Accordingly, it is estimated that 2000 to 4000 Pacific Yew trees, which are 50 to 250 years old, must be cut down to yield 1 kg of taxol. Consequently, many companies and research institutes are attempting to develop alternative processes for producing taxol.
As a substitute for extracting the alkaloid from rare plant sources, there can be mentioned a process for extracting the desired alkaloid from cultured cells from the objective plant tissues grown in an artificial growth medium, but no example of a successful production and/or isolation of taxol from cultured cells of the plant belonging to the genus Taxus has been described in any literature. For example, M. A. Zenkteler et. al. (Acta. Soc. Bot. Pol., 39 (1): pp. 161-173, (1970)) disclose the formation of calli from female gametophytes of Taxus baccata LINN., but there is no description of whether or not the disclosed callus formation protocols can be applied to other species of the genus Taxus, and no description of whether or not the calli produce any alkaloid. Also there is no description teaching such matters. Nevertheless, research into the development of processes for producing taxol by cell culture has not been abandoned, since cell culture is still thought to be feasible as a process for producing same.
Therefore, the object of the present invention is, to provide a process for efficiently producing taxol by cell culture of Taxus plant.
DISCLOSURE OF THE INVENTION
To attain the object described above, the present inventors have made intensive research into callus initiation from tissues of plants belonging to the genus Taxus and into proliferation of the callus cells. As a result, it has been found that cultured cells having a high taxol content can be efficiently obtained by culturing specific tissues originating from specific species belonging to the genus Taxus, whereby the present invention was achieved.
Namely, according to the present invention, there is provided a process for producing taxol by culturing cells originating from tissues of plants belonging to the genus Taxus, which comprises: suitable for inducing a callus, to thereby induce a callus; medium suitable for proliferating suspension culture cells; and c).
Also provided are a process for inducing and proliferating the callus using some of the above-described steps, and the callus or suspension culture cells containing at least taxol as an alkaloid.
BEST MODE OF CARRYING OUT THE INVENTION
The phrase "plants belonging to the genus Taxus", as used in the present invention, means plants belonging to the genus Taxus of the family Taxaceae, and suitable for the object of the present invention (i.e. production of taxol). Typical examples thereof include, but are not restricted to, Pacific Yew (Taxus brevifolia NUTT.) and Japanese Yew (Taxus cuspidata SIEB. et ZUCC.).
According to the present invention, living tissues are prepared from specific tissues of these plants in a first stage. Consequently, as a piece of plant material put into culture
REFERENCES:
patent: 5019504 (1991-05-01), Christen et al.
S. H. Mantell et al. (1985) Principles of Plant Biotechnology pp. 91-94.
Christen et al., "Proceedings of the Eightieth Annual Meeting of the American Association for Cancer Research", 30 566(2252)(May 24-27, 1989).
Hibi Masaaki
Ohashi Hiroaki
Saito Koji
Tahara Makoto
Campell Bruce R.
Fox David T.
Nippon Steel Corporation
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