WO1999027802A1

WO1999027802A1 - HEALTH BEVERAGE CONTAINING THE EXTRACT OF $i(PHELLINUS LINTEUS)

Info

Publication number: WO1999027802A1
Application number: PCT/KR1997/000246
Authority: WO
Inventors: Nam-Doo Hong; Man-Woo Han; Jae-Kuk Yoo
Original assignee: Han Kook Sin Yak Pharm. Co., Ltd.
Priority date: 1997-11-28
Filing date: 1997-11-28
Publication date: 1999-06-10

Abstract

According to the present invention, a health beverage containing the extract of Phellinus linteus as main ingredient and manufacturing method thereof are disclosed and regular drinking of these beverages can reduce the incidence rate of cancers.

Description

Health beverage containing the extract of Phellinus linteus

Detailed Description of the invention

The present invention concerns with a health beverage containing the

extract of Phellinus linteus as main ingredient and manufacturing

method thereof.

Basidiomycetes have been used for a long period of time as an

oriental folk herbal medicine. Especially, the basidiomycetes-produced

polysaccharides were known to have an excellent anticancer activity with

less side effects (toxicologically safe) and an activity of potentiating the

immune system. Two methods were known for the use of

basidiomycetes^ 1) The use of fruiting body of basidiomycetes by

collecting directly from natural resources or planting the fruiting body

and 2) The use of mycelia after separating mycelia from fruiting body

and cultivating the mycelia. The method of directly collecting naturally

formed fruiting body causes the depletion of natural resources and

ecological disruption. Therefore, artificially cultivating method has been

employed, however, this method has some disadvantages; 1) slow

growing rate, and 2) bacterial contamination.

In order to solve the above mentioned problems, the inventors

developed the method for cultivating mycelia of Phellinus linteus after a

long-term study and obtained patents or are applying for patents (Korea Patent Publication No. 92-1367, Polysaccharides and a method of their

production; Korea Patent Publication No. 92-1194, Manufacturing method

of mycelia of Phellinus linteus,' Korea Patent Publication No. 97-9150,

Artificial liquid cultivation of mycelia of Phellinus linteus and

manufacturing method of compounds having anticancer-immune activity!

Korea Patent Laid Open Publication No. 97-15743, Manufacturing method

of polysaccharides having anticancer-immune activity isolated from

Phellinus linteus,^' and Korea Patent Laid Open Publication No. 97-1531,

A new strain of Phellinus linteus producing anticancer-immune activity).

The polysaccharides with potentiating immune system could be

obtained after purification of the extract of Phellinus linteus (Korea

Patent Publication No. 92-1367; Korea Patent Laid Open Publication No.

97-15743), and this could be used in this invention.

The extract and polysaccharides ("Phellinus linteus extract'")

obtained from mycelia of Phellinus linteus by above mentioned method

have the following pharmacological activities; 1) potentiation of immune

system when combination chemotherapy was employed to treat

gastrointestinal cancer [The New Korean Medical Journal, Vol. 39, No. 11,

November , 1996] and hepatoma, and after cancer dissection

surgery [Journal of Korean Cancer Association, Vol.29, No. 3, June,1997

and 2) detoxification activities. Therefore, the "Phellinus linteus extract"

has been used as a medicine in the form of powder or capsule. Since the modern population are exposed to a variety of air pollutants

and carcinogenic compounds, the incidence rate of cancer is very high.

Not only the cure rate of cancer is low, but also the cancer patients are

suffering from severe pain and survival rate is not high, the best

method is to prevent cancer.

Now, the present inventors developed health beverages containing

the extract of Phellinus linteus as main ingredient, after long-time

study. Regular drinking of these beverages can reduce the incidence

rate of cancer and the taste and color of these beverages are nearly the

same with those of common beverages, and no precipitates are formed.

The present invention covers the following beverages containing the

extract of Phellinus linteus,' fruit juices, alcoholics, coke, milk, coffee,

tea, and common alcoholic and non-alcoholic beverages.

The juices contain orange juice, tomato juice, apple juice, strawberry

juice, pear juice, and other common fruit juices. The concentrated

(100%) and diluted (by adding water) juices are also covered by the

present invention.

The coffees contain dried pre-mix and canned or bottled instant

coffee.

The teas contain dried pre-mix and caned or bottled instant tea of

green tea and red tea.

The present invention also covers flavored beverages. The flavored beverages are nonalcoholic beverages containing orange-, grape-, lemon-,

and strawberry-flavored beverages.

The present invention also covers alcoholic beverages; beers and

wines, and apple- and lemon-containing alcoholic beverages, and other

lo -alcoholic beverages.

1-5,000 mg (preferably 5-1,000 mg) of the extract of Phellinus linteus

could be added to 100 ml of beverages.

1 - 5,000mg (preferably 5 - l,000mg) of the extract of Phellinus

linteus could be added to l,000mg of dried pre-mix of coffee or tea.

The present healthy beverages where the extract of Phellinus linteus

was added to common beverages could be sterilized, if necessary.

The following examples and experiments explain concisely the present

invention.

Example 1.

Culture of Phellinus linteus strain, FERM BP-2639(Deposited to Japan

Institute of Microorganism).

The 1 litter of culture medium containing 50 g of glucose, 10 g of

pepton, 10 g of yeast extract solution, and 0.5 g K2HP0 was employed

to culture the Phellinus linteus strain. The 1 litter of main culture

medium containing 10 g of soluble starch, 50 g of glucose, 5 g of yeast extract solution, 10 g of corn steep powder, 0.5 g of K2HPO 0.3 g of

MgSO , and trace elements of MnCb, FeSO₄, and ZnSO was used.

Both the seed culture medium and main culture medium were adjusted

to pH 5.0 before sterilization.

After sterilization of 500 ml flask containing 50 ml of seed culture

medium for 20 min at 121 °C, Phellinus linteus strain, FERM BP -2639[

Deposited to Japan Institute of Microorganism] on the

potato-glucose-agar slant was collected with small portion of the slant

and finally cultured with agitation for 3 days at 28 °C ("the first seed

culture medium"). The jar fermenter containing 3/ of seed culture

medium was sterilized for 30 min at 121 °C and 50 ml of the above

mentioned ("the first seed culture") medium was added to the jar

fermenter, and then were cultured for 3 days at 28 °C ("the second seed

culture medium").

6 litter of "the second seed culture medium" was added to 500 litter

of presterilized fermenter containing 300 litter of main culture medium

and the mixture was cultured with agitation at 28 °C with 300 1/min of

aeration and 100-200 rpm of stirring. The above mixture was added to a

presterilized fermenter containing 3000 litter of main culture medium and

the mixture was cultured for 3 days with 3000 1/min of aeration and

100-200 rpm of stirring. The above procedures were repeated to obtain Example 2

Extraction of the compounds having the activities of anticancer and

potentiating immune system. The mycelia obtained from example 1

were extracted and purified by the following procedures. 500 ml of

distilled water were added to 100 g of the mycelia and extracted twice

for 2 h of boiling at 90-100 °C. After discarding the cake, the extracted

water layer was collected. The extracted water layer was concentrated

to 100 ml under vacuum and 3 volumes of 95% ethanol were added.

After overnight standing, centrifugation was performed for 30 min at

3000 rpm. 100 ml of water was added to dissolve the precipitate and

the mycelia having molecular weight of less than 8000 was obtained

using semipermeable membrane for 3 days. The resultant high

molecular weight component containing polysaccharides, 3 g, was

obtained after freeze drying at -70 °C.

Example 3.

Extraction of crude polysaccharides

10-20 volumes of water were added to the culture medium obtained

from example 1 and the mixture was boiled for 1-2 hr at 50-100 °C.

After centrifugation, the filtrate was concentrated using

Reduced-pressure Film Evaporator. Crude polysaccharides were obtained

either freeze drying or spray drying from the above mentioned concentrated solution.

Experiment 1.

Acute toxicity test

The extract of Phellinus linteus obtained from example 2, 10,000

mgAg, was injected intraperitoneally to 4- week old ICR mouse (n-10).

These were no significant findings after 14 days.

Experiment 2.

Anticancer activity

The anticancer activity against sarcoma 180 of the extract of

Phellinus linteus obtained from example 2 was performed in 4-week-old

ICR mice (n=10). At seventh day after implantation of sarcoma 180 to

ICR mice peritoneum, 1 x 10⁶ cells were implanted subcutaneously into

the mice left groin. The extract of Phellinus linteus was dissolved in

normal saline injectable solution and filtered using 0.45 m filter. And

then 100 mgAg was injected intraperitoneally everyday for 10 days from

24 h after implantation into the groin. At 30th day after implantation

into the groin, the cancer mass was isolated and the percentages of

cancer growth inhibition were measured. The results are listed in Table

1. As listed in Table 1. the anticancer activity of the extract of

Phellinus linteus was approximately 71.5%. Table 1. Anticancer activity of the extract of Phellinus linteus against

sarcoma 180

It has been found that the extract of Phellinus linteus had an

excellent activity against gastrointestinal cancer (New Med. J. Korea, 39,

1996) and had potentiating immune system (J. Kor. Cancer Assoc, 29,

1997), and had an excellent effect against B16 (Int. J. ImmunopharmacoL,

18, 295-303, 1996)

Above data indicates that the extract of Phellinus linteus has little

toxicity and has anticancer activity. Therefore, the extract could be

used as health food to prevent and treat the cancer

The present invention will be illustrated in detail by the preparation

examples below.

Preparation Example 1

To each 100ml of undiluted orange juice was added each lOmg, 20mg, 30mg, 40mg, 50mg and lOOmg of extract of Phellinus linteus P

Ex.) and each mixture was dissolved with well stirring at ambient

temperature and filled in each bottle of 100ml and sterilized. The color,

taste and precipitate of each orange juice prepared was compared with

those of original orange juice.

The results are as follows.

Preparation Example 2

To each 100ml of milk was added each lOmg, 20mg, 30mg, 40mg,

50mg and lOOmg of extract of Phellinus linteus(¥ Ex.). Each mixture

was dissolved with well stirring at ambient temperature, filled in each

bottle of 100ml and was sterilized. The color, taste and precipitate of

each milk prepared was compared with those of original milk.

The results are as follows.

Preparation Example 3

To each undiluted cola liquid was added each lOmg, 20mg, 30mg,

40mg, 50mg and lOOmg of extract of Phellinus linteus(PL Ex.). Each

mixture was diluted with purified water, controlled sugar content with

sugar and filled in each bottle of 100ml. CO2 gas was filled in each

mixture and was sealed. Each mixture was sterilized in a conventional

preparation method of cola to prepare each cola. The color, taste and

precipitate of each cola prepared was compared with those of original

cola.

The results are as follows.

Preparation example 4

To each 100ml of beer was added lOmg, 20mg, 30mg, 40mg, 50mg

and lOOmg of extract of Phellinus linteus( PL Ex.) and each mixture

was dissolved with well stirring and filled in each bottle of 100ml. To

the mixture was filled CO2 gas to be controlled to the conventional CO2

gas pressure of beer and was sterilized in a conventional preparation method of beer. The color, taste and precipitate of each beer prepared

was compared with those of original beer.

The results are as follows.

Preparation Example 5

To each 100ml of portwine of 12%(v/v) was added lOmg, 20mg,

30mg, 40mg, 50mg and lOOmg of extract of Phellinus linteus(FL Ex.)

and each mixture was dissolved with well stirring. The color, taste and

precipitate of each portwine was compared with those of original

portwine.

The results are as follows.

Preparation Example 6

To each 100ml of milk was added 5mg and l,000mg of extract of

Phellinus linteus(PL Ex.) and each mixture was dissolved with well

stirring and filled in each bottle of 100ml and pasteurized. The color,

taste and precipitate of each milk was compared with those of original

milk.

The results are as follows.

Preparation Example 7

To each solution of l.Og of each dried granulized coffee extract in

each 80ml of hot water was added lOmg, 20mg, 30mg, 40mg, 50mg and

lOOmg of extract of Phellinus linteus(PL Ex.) and each mixture was

dissolved with well stirring to make each black coffee. The color, taste

and precipitate of each black coffee was compared with those of original

black coffee. The results are as follows.

Preparation Example 8

To each solution of 2.0g of dried granulized coffee extract in each

200ml of water was added lOmg, 20mg, 30mg, 40mg, 50mg and lOOmg

of extract of Phellinus linteus( PL Ex.) and each mixture was dissolved

with well stirring, filled in each can of 200ml, sealed and sterilized in a

conventional preparation method of canned coffee. The color, taste and

precipitate of each canned coffee was compared with those of original

can coffee.

The results are as follows.

Preparation Example 9

To each l.Og of dried granulized coffee extract was added lOmg, 20mg, 30mg, 40mg, 50mg and lOOmg of extract of Phellinus linteus(FL

Ex.) and each mixture was well mixed with stirring and was sealed in

each paper box. They are stored for 6 months at ambient temperature.

Each box was dissolved in each 80ml of hot water and the color, taste

and precipitate of each coffee was compared with those of original

coffee.

The results are as follows.

Preparation Example 10

To each solution of 2.0g of dried granulized green tea extract in

each 200ml of water was added lOmg, 50mg, lOOmg, 500mg, l,000mg

and 3,000mg of extract of Phellinus linteus(PL Ex.) and each mixture

was dissolved with well stirring, filled in each can of 200ml, sealed and

sterilized in a conventional preparation method of canned tea. The color,

taste and precipitate of each can tea was compared with those of

original canned green tea. The results are as follows.

Preparation Example 11

To each l.Og of dried granulized red tea extract was added lOmg,

20mg, 30mg, 40mg, 50mg and lOOmg of extract of Phellinus linteus PL

Ex.) and each mixture was well mixed with stirring and was sealed in

each paper box. They were stored for 6 months at ambient temperature.

Each box was dissolved in 80ml of hot water and the color, taste and

precipitate of each red tea was compared with those of original red tea.

The results are as follows.

The present extract of Phellinus UnteusiPL Ex.) can be mixed with powder of dried coffee extract or dried granuUzed coffee extract and the

mixture can be dissolved in hot water; or can be dissolved with coffee

extract to prepare an instant coffee solution.

The present extract of Phellinus linteus(PL Ex.) can be mixed with

powder of dried tea extract or dried granulized tea extract and the

mixture can be dissolved in hot water; or can be dissolved with tea

extract to prepare an instant tea solution.

Any other kinds of fruit juice of the present invention than orange

juice can be prepared in accordance with the Example 1. It is evident

that they come under the scope of the present invention.

Claims

What we claim is-

1. A health beverage containing extract of Phellinus linteus as main

ingredient and other ingredients commonly used in common beverage

prepared by conventional preparing method of beverage with extract of

Phellinus linteus and other ingredients used in common beverage.

2. A health beverage of the claim 1 containing 1.0 - 5,000mg of

extract of Phellinus linteus as main ingredient in 100ml of the healty

beverage and other ingredients commonly used in common beverage

prepared by conventional preparing method of beverage with extract of

Phellinus linteus and other ingredients used in common beverage.

3. A health beverage of the claim 1 containing extract of Phellinus

linteus as main ingredient and other ingredients commonly used in

common beverage wherein other ingredients commonly used in common

beverage are selected from the group consisting of juice, flavor

beverage, milk, alcoholic beverage, cola, coffee and tea.

4. A health beverage of the claim 1 containing extract of Phellinus

linteus as main ingredient and other ingredients commonly used in coffee

beverage wherein coffee beverage is an instant coffee drink containing

extract of Phellinus linteus as main ingredient and other ingredients

commonly used in instant coffee drink; or a dried coffee mixture

containing extract of Phellinus linteus as main ingredient and dried

powder or dried granulated coffee extract in mixed state and when drinking the coffee beverage, the coffee mixture is diluted in water.

5. A Manufacturing method of health beverage containing extract of

Phellinus linteus as main ingredient and other ingredients commonly

used in common beverage prepared by conventional preparing method of

beverage with extract of Phellinus linteus and other ingredients used in

common beverage.