Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Reexamination Certificate
1999-12-09
2001-08-14
Lilling, Herbert J. (Department: 1651)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
C536S119000, C536S123100, C435S170000, C435S253100, C435S863000
Reexamination Certificate
active
06274356
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a carbohydrate complex extracted from
Mycobacterium tuberculosis
, which has an anticancer activity, and to a process for the preparation thereof.
BACKGROUND OF THE INVENTION
It is generally known that the anticancer activity of
Mycobacterium tuberculosis
is attributable to active agents in the cytoplasmic membrane thereof, particularly the polysaccharide and lipid derivatives.
For instance, Azuma et al. succeeded in isolating N-acetylmuramyl-L-alanyl-D-isoglutamin(MDP) which is an active component of
M. tuberculosis
[Azuma, L. et al.,
J. Bact
., 96, 1885-1887(1968)].
Barnes et al., on the other hand, reported that lipoarabinomannan promotes the production of cytokin [Barnes, P. F. et al.,
J. Immunol
., 149, 541-547(1992)]. Chung et al. isolated from
M. tuberculosis
a substance having an anticancer activity and used this substance, designated as Tubercin-3, in treating terminal-cancer and leprosy patients as well as laboratory animals suffering from tumoral symptoms [Chung, T. H.,
J. Korean Med. Ass
., 17, 427-431(1974); Chung, T. H. et al.,
Yonsei Med. J
., 17, 131-135(1976)].
On the other hand, a person infected by
M. tuberculosis
bacilli normally exhibits granulomatous inflammation, and if the infected site is lung, cavity formation progresses together with an inflammatory reaction, induced by secretion of proteins and glycolipids originally present in the cell wall of
M. tuberculosis
. This suggests that, in case high molecular-weight polysaccharides or proteins are used in cancer immunotherapies, it may be difficult to suppress such adverse side effects as undesirable immuno-responses and uncontrollable inflammatory reactions.
SUMMARY OF THE INVENTION
Accordingly, it is an object. of the invention to provide a carbohydrate complex extracted from
Mycobacterium tuberculosis
which has a high anticancer activity and at the same time causes essentially none of the undesirable side-effects mentioned above.
It is another object of the invention to provide a process for the preparation of said carbohydrate complex.
In accordance with one aspect of this invention, there is provided a carbohydrate complex, which is a mixture of polysaccharides extracted from
Mycobacterium tuberculosis
, characterized in that: the polysacchricdes have the molecular weight of below 7,000 and are derived from monosaccharides consisting essentially of mannose, arabinose, glucose and galactose, the monosaccharides forming straight-chain and/or side-chain glycosidic bonds.
In accordance with another aspect of the invention, there is provided a process for the preparation of the carbohydrate complex, comprising the steps of:
(a) culturing
Mycobacterium tuberculosis
in a medium, heating the culture diluted with water at a temperature ranging from 50 to 150° C. under a pressure ranging from 10 to 30 psi and removing bacterial residues from the heated culture solution;
(b) adding an inorganic salt to the remaining solution to form a precipitate and removing the precipitate from the resulting solution;
(c) dialyzing the remaining solution to obtain a dialysis product;
(d) adding an alcohol to the dialysis product to obtain a precipitate, washing the precipitate with an alcohol-ether mixture and extracting the washed, precipitate with an organic solvent; and
(e) purifying the extract to obtain the carbohydrate complex.
REFERENCES:
patent: 4329452 (1982-05-01), Maruyama
patent: 4394502 (1983-07-01), Maruyama
patent: 4746511 (1988-05-01), Kobatake et al.
Chung Chong-Chan
Chung Tai-Ho
Anderson Kill & Olick PC
Lilling Herbert J.
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