Tumor necrosis factor inducing substance derived from acid-fast

Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Carbohydrate doai

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424 92, 536 552, A61K 3574, A61K 3700, C12P 100

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active

049007246

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD:

The present invention relates to an amphipathic substance having tumor necrosis factor (TNF) inducing activity which is obtained from acid-fast bacteria by means of extraction. The amphipathic substance according to the present invention is novel and may effectively be employed as a medicine, e.g., an antitumor agent.


BACKGROUND ART:

Lipoteichoic acid (hereinafter referred to as "LTA"), lipopolysaccharide (hereinafter referred to as "LPS") and the like have heretofore been known as amphipathic substances which are extracted from, for example, the whole cell of Gram-positive bacteria and Gram-negative bacteria or a fraction including the cell wall and cytoplasmic membrane of such bacteria (such fraction being referred to as a "cell envelope fraction", hereinafter).
In 1975, Old et al, reported a tumor necrosis factor (TNF) which is a glycoprotein that specifically attacks only tumor cells without damaging normal cells (Proc. Natl. Acad. Sci. U.S.A., 72, 3666). According to this report, it is possible to induce TNF in serum by administering live BCG or the like into the vein of a mouse and administering LPS one to two weeks later. However, the lethal toxicity of LPS is extremely high; for example, if 0.8 .mu.g/head or more of LPS is administered to mice sensitized with BCG, the greater part of the mice will die within six hours. Accordingly, employment of LPS itself as a medicine such as a cancer controlling agent involves clinical problems.


DISCLOSURE OF THE INVENTION:

The present invention relates to an amphipathic substance having TNF inducing activity which is obtained by extraction from acid-fast bacteria which belong to a range which is different from those to which Gram-positive bacteria and Gram-negative bacteria belong. The amphipathic substance according to the present invention is distinguished from LTA in that the glycerol content in the novel substance is less than the limit of detection and the organic phosphorus content therein is exceedingly lower than that in LTA. Further, the amphipathic substance of the present invention also differs from LPS in that the novel substance contains no hydroxy fatty acid such as 3-hydroxy myristic acid. The chemical composition of the amphipathic substance according to the present invention is as follows (mg/mg; mole/mg for organic phosphorus):


______________________________________ hexose 0.25-0.55 pentose 0.10-0.17 fatty acid 0.03-0.16 amino acid 0.07-0.17 amino sugar <0.03 (generally, 0.001-0.03) organic phosphorus 0.3-1.7 glycerol not detected ______________________________________ of mannose, while the greater part of the pentose consists of arabinose. The fatty acid contains palmitic acid, stearic acid and tuberculostearic acid as principal components, and no hydroxy fatty acid is detected. Chief amino acids are glutamic acid, aspartic acid, leucine, threonine, glycine, alanine, serine and valine, and a trace of diaminopimelic acid is detected. The amino sugar consists mainly of glucosamine and contains substantially no muramic acid.
The acid-fast bacteria having the novel amphipathic substance of the present invention are aerobic bacteria which show acid-fast property in cell staining and have in their cell walls a large amount of fatty acids, particularly mycolic acid which is an .alpha.-branched, .beta.-hydroxy higher fatty acid. Practical examples of such acid-fast bacteria include species and strains which belong to genera Mycobacterium, Nocardia, Rhodococcus, Gordona and Corynebacterium. The number of carbon atoms of the mycolic acid which is a constituent component characteristic of the cell walls of these acid-fast bacteria is different for each genus: 60-90 for genus Mycobacterium: 30-70 for genera Nocardia and Rhodococcus: 60-80 for genus Gordona: and 30-40 for genus Corvnebacterium.
Practical examples of the acid-fast strains which may be employed in the present invention include those mentioned below. However, the present invention is not necessarily limited thereby.
Mycobacterium tubercul

REFERENCES:
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S. Haskill et al., "Effect of C. Parvum on Intratumor Immunity to the T1699 Mammary Adenocarcinoma", The Journal of Immunology, vol. 125, No. 1, 1980, pp. 454-458.

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