Proteoglycan(G009) effective in enhancing antitumor immunity

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Glycoprotein – e.g. – mucins – proteoglycans – etc.

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Details

530322, 435 712, 435 701, A61K 3570, C07K 1437

Patent

active

055854672

DESCRIPTION:

BRIEF SUMMARY
BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a saccharide analysis chromatogram of proteoglycan G009 according to the present invention.
FIG. 2 is an amino acid chromatogram of proteoglycan G009 according to the present invention.
FIG. 3 is an IR analysis diagram of proteoglycan G 009 according to the present invention.
FIG. 4 is a microphotograph showing the mycelia of a strain IY 009 according to the present invention.
FIG. 5 is a diagram drawn to compare the specificity of a strain IY 009 according to the present invention.


DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a proteoglycan effective for promotion of antitumor immunity.
In order to search for a new proteoglycan which has the effect of promotion antitumor immunity, the present inventors collected a large number of basidiomycetes and attempted to separate a proteoglycan produced by those basidiomycetes. In sequel thereto, it was found that the basidiomycete which belongs to the genus of ganoderma, picked up in the area of the Dooryoon Mountains located in the South Cholla Provine, produces a proteoglycan which has the effect of promoting antitumor immunity when cultured in a proper culture medium. After the produced proteoglycan was separated and its physicochemical and biophysical properties was examined, it was designed as proteoglycan G009 which has the effect of promoting antitumor immunity and the strain which produces it was designated as Ganoderma lucidum IY 009 (a number of deposition to the Korean Species Association is KFCC-107090 dated Oct. 5, 1990).
Although researches have been conducted in such medicinal components as antibacterial, hallucinative, toxicant and cholesterol-lowering components from the basidiomycetes belonging to a higher fungus, earnest study is now given to an antitumor component and an immunity improvement action, and so basidiomycetes are rising in importance. Thus, those strains which secrete such matter as contains an antitumor component and improves an immunity function was separated and cultured in liquid to make a study of its pharmacological effect.
During the study, a mutant strain which has an antitumor effect and an immunity improvement function that are more excellent than the species belonging to the genus of Ganoderma which has been identified until now was separated and an in-liquid mycelia culture method was established and then the antitumor effect and immunity improvement function of those components extracted from the cultured mycelia were confirmed.
The present invention will now be described in detail according to the following embodiment and experimental examples.


EXAMPLE 1

Separation of proteoglycans
1) Strain:
The Ganoderma lucidum starins were collected in the area of the Dooryoon Mountains located in the South Cholla Province and identified.
2) Preserved culture medium:
An inclined culture medium for potato dextrose agar (PDA); 39 g of potato dextrose culture medium (Difico, USA) was made in to 1 liter by dissolving it in distilled water and it was made into an inclined culture medium after high-pressure sterilization for 20 minutes at 121.degree. C.
A culture medium for in-liquid culture:
50 g of glucose, 20 g of peptone, 0.87 g of KH.sub.2 PO.sub.4, 0.5 g of MgSO.sub.4.7H.sub.2 O, 10 mg of FeCl.sub.2.6H.sub.2 O, 7 mg of MnCl.sub.2.4H.sub.2 O, 10 mg of ZnSO.sub.2.5H.sub.2 O and 4 mg of ZnCl were made into 1 liter by adding distilled water thereto. It was sterilized for 20 minutes at 121.degree. C. by adjusting the pH value to 5.5.
3) Culture:
The strains kept in custody were transplanted into the inclined culture medium for PDA and grown for 7 days at 25.degree..+-.1.degree. C. Then, the grown mycelia were separated in an aseptic way and put into 100 ml of culture medium for in-liquid culture and triturated with a microblender for 15 seconds. It was moved into a 500 ml-triangular flask and shake-cultured for 10 days until the mycelium formed a mature mycetome with a diameter of about 5 mm by 180 rpm at 25.degree..+-.1.degree. C.
After the mycetome was tritur

REFERENCES:
Lee et al., "Pharmacological, Toxicological Studies of Antitumor Polyaaccharides Obtained from Ganodmora lucidum IH 009", Kor. J. Appl. Microbiol. Technol. vol. 22 (2) pp. 182-89 (1994) pp. 182 & 187 Applied as Exhibit A.
Shimura et al, Japan J. Pharmacol., vol. 33, pp. 403-408 (1983).
Suzuki et al, Chem. Pharm. Bull., vol. 37(2), pp. 410-413 (1989).

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