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Cancer Patent Abstract
An extract of the Minari plant containing a component which prevents
cancer and cancer metastasis. A Minari extract which is useful for
the prevention of cancer and cancer metastasis induced by urethane
(EC (ethylcarbamate)), which is a carcinogen present in fermented
foods or alcoholic beverages, or by NNK[{(4-N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-buta
none)}], which is a carcinogen present in cigarette smoke.
Cancer Patent Claims
The invention claimed is:
1. A method of reducing the incidence of lung cancer comprising
administering to an individual an effective amount of a Minari extract,
which is prepared by extracting a Korean native dol-minari of Oenanthe
javanica plant in hot water, and which comprises a component which
reduces expression of cyclin D1.
2. A method according to claim 1, wherein the lung cancer is cancerwhich
can be induced by urethane or NNK [{4-N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-butanone}].
3. A method of reducing the incidence of lung cancer comprising
administering to an individual an effective amount of a composition
comprising a pharmaceutically acceptable carrier and a Minari extract,
wherein the Minari extract is prepared by extracting a Korean native
dol-minari of Oenanthe javanica plant in hot water.
4. A method according to claim 3, wherein said composition is in
a form selected from the group consisting of powder, liquid, tablets,
capsules, and pellets.
5. A method of reducing the incidence of lung cancer comprising
administering to an individual an effective amount of a Minari extract
comprising a component which is useful as a therapeutic agent for
treating cancer in a specimen, wherein the cancer is lung cancer,
wherein the cancer is cancer induced by urethane or NNK, and wherein
the extract is prepared by extracting a Korean native dol-minari
of Oenanthe javanica plant in hot water.
6. A method of reducing the incidence of lung cancer comprising
administering to an individual an effective amount of a Minari extract
prepared by extracting plant material of a Korean native dol-minari
of Oenanthe javanica plant in hot water, comprising components which
reduce expression of cyclin D1; reduce expression of proliferative
cell nuclear antigen (PCNA); reduce NNK activation and thereby reduce
lung cancer multiplicity; and reduce the expression level of PCNA.
Cancer Patent Description
TECHNICAL FIELD
The present invention relates to an extract of the Minari plant
containing a component which prevents cancer and cancer metastasis.
More particularly, the present invention relates to a Minari extract
which is useful for the prevention of cancer and cancer metastasis
induced by urethane (EC (ethylcarbamate)), which is a carcinogen
present in fermented foods or alcoholic beverages, or by NNK[{(4-N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-buta
none}], which is a carcinogen present in cigarette smoke.
BACKGROUND ART
Urethane and NNK are known to be involved in the occurrence of
various types of cancer.
Urethane causes lung cancer and liver cancer in mice in a dose-dependent
manner. The virtually safe dose (VSD) of urethane in humans has
been calculated to be 1.8.times.10.sup.-4 mg/kg body weight for
lung cancer and 7.2.times.10.sup.-5 mg/kg body weight for liver
cancer (K. Inai et al., Jpn. J. Cancer Res. 82, pp. 380-385, April
1991).
The offspring of pregnant mice, treated with urethane during gestation
and lactation, had a significantly increased incidence of embryonic
tumors, lung cancer, and ovarian cystadenomas via placental transmission,
as well as endometrial hyperplasias and uterine hemangiomas in the
pregnant parent mice (T. Nomura, Cancer Research, 33, pp. 1677-1683,
1973, July).
When administered per os, urethane causes lung cancer, lymphomas,
hepatoma, stomach papillomas, sebaceous carcinomas, mammary tumors,
squamous-cell tumors, leukemia, and mesenchymal tumors in mice (IARC
MONOGRAPHS ON THE EVALUATION OF THE CARCINOGENIC RISK OF CHEMICALS
TO MAN, vol. 7, pp. 111-131, the views of two IARC Working Groups
on the Evaluation of the Carcinogenic Risk of Chemicals to Man which
met in Lyon, 4-11 Feb. 1974 and 18-24 Jun. 1974).
Moreover, regulatory limits are implemented on urethane in Canada
because urethane is produced through alcoholic fermentation and
is contained at a certain level in brews such as wine (Ethyl Carbamate
in Alcoholic Beverages and Fermented Foods, ACS Symposium Series
No. 484, Food Safety Assessment, ed. by John W. Finley et al., pp.
419-428, published in 1992 by American Chemical Society; Identification
of Volatile Constituents from Grapes, J. Agric. Food Chem., vol.
24, no. 2, pp. 329-331, 1976; RATIONALE FOR THE ESTABLISHMENT OF
GUIDELINES TO LIMIT ETHYL CARBAMATE LEVELS IN ALCOHOLIC BEVERAGES,
BUREAU OF CHEMICAL SAFETY, FOOD DIRECTORATE HEALTH PROTECTION BRANCH,
HEALTH & WELFARE CANADA, pp. 1-8; Ethylcarbamate in Fermented
Beverages and Foods, Cornelius S. Ough, J. Agric. Food Chem. vol.
24, no. 2, pp. 323-327).
NNK is a potential carcinogen present in cigarette smoke (Djordijevic,
M. V. et al., A comparison of selected components in the mainstream
smoke of the leading U.S. and Japanese cigarettes. In: Proceedings
of the CORESTA smoke and Technology Meeting, pp. 200-217, Nov. 3-8,
1996).
Among other types of cancer, the incidence of lung cancer has continued
to increase in both men and women. The occurrence of lung cancer
is significantly related to smoking and alcohol consumption. The
International Agency for Research on Cancer has indicated synergistic
consumption of smoking and alcohols, suggesting that NNK and urethane
which have been totally taken in through smoking and through intake
of alcoholic beverages are a cause for lung cancer. It has been
indicated that smoking directly contributes to 45% of cancer deaths
in men and 21.5% in women. It appears that death from lung cancer
has replaced cardiac diseases as a main cause of death among smokers
in the United States.
There is a need for a dietary ingredient for preventing carcinogenesis
due to smoking and alcohol intake.
DISCLOSURE OF THE INVENTION
The present invention relates to a dietary ingredient of Minari
extract for preventing occurrence and subsequent metastasis of cancer.
Minari, a plant of Family Umbelliferae, is grown and cultivated
in damp soil or in rice paddies, and traditionally known as a detoxifying
agent. It is known to very effectively detoxify alcohol toxicity
by promoting excretion of alcohol.
In Chinese herbal medicine, Minari is used to treat jaundice, gastrointestinal
diseases, neurosis, and obesity. However, it has not previously
been found that Minari prevents occurrence of cancer and even obstructs
metastasis of cancer. The inventor has found that a dietary ingredient
of Minari extract obstructs activation of potential precarcinogens,
eliminates potential free radicals which are produced through metabolism
of carcinogens, and directly eliminates free radical generators
among chemical carcinogens, and therefore that the Minari extract
is highly effective for cancer prevention. Thus, the present invention
was accomplished.
The present invention provides a Minari extract containing a component(s)
which prevents cancer and cancer metastasis.
According to one aspect of the present invention, the extract is
prepared by solvent extraction. According to another aspect of the
present invention, the extract is prepared by extraction in hot
water.
According to another aspect of the present invention, the composition
is selected from the group consisting of persicarin, hyperoside,
rutin, quercetin, chlorophyll, xanthophyll, isohamenetin, phenethylisothiocyanate,
phenalkylisothiocyanate, flavons, flavonols, and d-limonene.
According to still another aspect of the present invention, the
cancer is cancer which can be induced by urethane or NNK[{(4-N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-buta
none}]. According to a further aspect of the present invention,
the cancer is lung cancer or cancer via placental transmission or
embryonic tumors.
The present invention further provides a composition which prevents
cancer and cancer metastasis containing a Minari extract and a pharmaceutically
acceptable carrier.
According to one aspect of the present invention, the composition
is in a form selected from the group consisting of powder, liquid,
tablets, capsules, and pellets.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 demonstrates different levels of lung cancer multiplicity
induced in three experimental groups of A/J mice, taken after 4
months following a single gavage of different amounts of urethane,
as well as the suppressive effects of the Minari extract. Each of
the three experimental groups, which respectively received a single
administration of a 125, 250, or 500 mg/kg dose of urethane, was
divided into two subgroups: one of the subgroups received a normal
feed (urethane control group), while the other subgroup was allowed
to take, ad lib., a feed to which Minari extract was added at a
5% concentration (w/w) for 4 months (urethane+Minari). After the
experiment, the multiplicity rate of lung cancer (incidences of
cancer/mouse) was observed for each group.
FIG. 2 demonstrates different expression levels of the cyclin D1
oncogene in lung tissue in three experimental groups of A/J mice,
taken after 4 months following a single gavage of different amounts
of urethane, as well as the suppressive effects of the Minari extract.
Each of the three experimental groups, which respectively received
a single administration of a 125, 250, or 500 mg/kg dose of urethane,
was divided into two subgroups: one of the subgroups received a
normal feed (urethane control group), while the other subgroup was
allowed to take, ad lib., a feed to which Minari extract was added
at a 5% concentration (w/w) for 4 months (urethane+Minari). After
the experiment, the expression level of cyclin D1 was observed for
each group.
FIG. 3 demonstrates different expression levels of PCNA (proliferative
cell nuclear antigen) of the S-phase of the cell cycle in lung tissue
in three experimental groups of A/J mice, taken after 4 months following
a single gavage of different amounts of urethane, as well as the
suppressive effects of the Minari extract. Each of the three experimental
groups, which respectively received a single administration of a
125, 250, or 500 mg/kg dose of urethane, was divided into two subgroups:
one of the subgroups received a normal feed (urethane control group),
while the other subgroup was allowed to take, ad lib., a feed to
which Minari extract was added at a 5% concentration (w/w) for 4
months (urethane+Minari). After the experiment, the PCNA level in
the lung tissue was observed for each group.
FIG. 4 demonstrates 8-OH deoxyguanosine levels of DNA isolated
from lung tissue in three experimental groups of A/J mice, taken
after 4 months following a single gavage of different amounts of
urethane, as well as the suppressive effects of the Minari extract.
Each of the three experimental groups, which respectively received
a single administration of a 125 mg/kg (data not shown), 250 mg/kg,
or 500 mg/kg (data not shown) dose of urethane, was divided into
two subgroups: one of the subgroups received a normal feed (urethane
control group), while the other subgroup was allowed to take, ad
lib., a feed to which Minari extract was added at a 5% concentration
(w/w) for 4 months (urethane+Minari). After the experiment, the
8-OH deoxyguanosine level of DNA isolated from the lung tissue was
observed for each group.
FIG. 5 demonstrates different levels of lung cancer multiplicity
induced in four experimental groups of A/J mice, taken after 4 months
following a single intraperitoneal administration of NNK[{(4-N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-buta
none}], which is a potential carcinogen in cigarette smoke, as well
as the suppressive effects of the Minari extract. The four experimental
groups were: an NNK control group and three groups (receiving 4
mg of NNK) which were allowed to take, ad lib., Minari at three
different doses (i.e., feed to which Minari was added at 1.25, 2.5,
or 5% (w/w)).
FIG. 6 demonstrates expression levels of the cyclin D1 oncogene
in lung tissue in three groups: a control group to which water was
intraperitoneally administered and which was thereafter allowed
to take, ad lib., a normal feed (CTL) for 4 months; a group receiving
a single intraperitoneal administration of NNK (4 mg) and thereafter
allowed to take, ad lib., a normal feed for 4 months (shown as 0
in the figure); and a group receiving a single intraperitoneal administration
of NNK (4 mg) and thereafter allowed to take, ad lib., a feed to
which Minari extract was added (1.25%, w/w) for 4 months.
FIG. 7 demonstrates different expression levels of PCNA (proliferative
cell nuclear antigen) of the S-phase of the cell cycle in lung tissue
in three experimental groups: a control group (water); an NNK control
group; and a group receiving NNK-Minari (1.25%, w/w), as well as
the suppressive effects of the Minari extract. The NNK group received
a single administration of 4 mg of NNK. The group receiving NNK-Minari
received a single administration of NNK and thereafter a feed to
which Minari extract was added at a 1.25% (w/w) concentration. The
other group received a normal feed. All the groups were allowed
take, ad lib., their feeds for 4 months. After the experiment, the
PCNA level in the lung tissue was observed for each group.
BEST MODES FOR CARRYING OUT THE INVENTION
The term "Minari" as used in the present invention refers
to a plant belonging to Family Apiaceae (Umbelliferae), and typically
a Korean native dol-minari of Genus Oenanthe.
The term "Minari extract" refers to a liquid obtained
through extraction from a Minari plant (leaves, stem, and roots)
using below-described extraction solvents, usually deionized water,
as well as a solid including powder obtained by lyophilizing the
same.
A Minari extract can be prepared by a method which is widely used
in common. For example, Minari is washed, dried, and pulverized
first. Then, extraction is carried out by using various solvents.
In general, the extraction solvent is added in a 2 to 10.times.
weight to the Minari powder. Examples of extraction solvents include
water, ethanol, propanol, butanol, acetone, 1,3-butylene glycol,
ethyl acetate, hexane, methylene chloride, methanol, or ethyl acetate.
Typically, water is used to prepare a Minari extract. A mixture
of two or more kinds among the aforementioned solvents may be used.
Typically, a mixture of Minari extract and a solvent is subjected
to extraction with agitation using a magnetic stirrer or the like
at room temperature for 24 hours. For example, by using deionized
water, extraction is carried out for 24 consecutive hours at 70.degree.
C. Thereafter, the deionized water is separated, followed by lyophilization.
The lyophilized powder is used as Minari extract. The components
contained in the Minari extract prepared in the above manner may
be separated by a HPLC (high performance liquid chromatography),
which is well-known to the those skilled in the art, and the separated
components may be identified by an NMR technique (nuclear magnetic
resonance technique).
The inventor has confirmed through NMR spectroscopy that persicarin,
hyperoside, rutin, quercetin, chlorophyll, xanthophyll, isohamenetin,
phenalkylisothiocyanate, phenethylisothiocyanate, flavons, flavonols,
and d-limonene are contained in the Minari extract thus prepared.
Some of these natural substances are known as antioxidants, and
these components are considered to act in a synergistic manner to
prevent carcinogenesis. Accordingly, a Minari extract containing
these components can capture various free radicals such as superoxides
and hydroxy free radicals, thereby preventing lung cancer and metastatic
tumors induced by NNK (a potential carcinogen among other carcinogenic
chemicals such as polyaromatic hydrocarbons, aza-arenes, aromatic
amine, conjugated alkylaldehyde, formaldehyde, NO, urethane, and
nicotine ornitrosamine, etc.),urethane (ethyl carbamate or alkyl
carbamate in fermented foods and alcoholic beverages), or the like.
Alternatively, these components can obstruct the expression of various
oncogenes.
The Minari extract may be used alone or in combination with Ostericum
coreanum.
The Minari extract can be used as dried powder, an additive to
foods, or a drink, and in formulations such as capsules, tablets,
or pellets fabricated by using a common binder. The detailed methodology
concerning the present invention will be described in the examples
below, which are not intended to limit the scope of claims.
The present invention is based on work directed to therapies using
Minari extract in various dosage forms, against, for example, a
carcinogen present in fermented foods or alcoholic beverages and/or
a potential carcinogen(s) present in cigarette smoke. However, the
inventor and the applicant contemplate that the present invention
can be applied to other types of cancer and metastatic cancer (including
cancer caused by other carcinogens). Furthermore, experimental schemes
and its protocols can be altered for practice.
The effects of preventing cancer and cancer metastasis provided
by the Minari extract of the present invention can be confirmed
by examining whether or not diets containing Minari extract at various
concentrations affect the expression level of the cyclin D1 gene
in the lung tissue of mice which have been treated with urethane.
The effects of preventing cancer and cancer metastasis provided
by the Minari extract of the present invention can be confirmed
by examining whether or not diets containing Minari extract at various
concentrations affect the expression level of PCNA (proliferative
cell nuclear antigen) in the lung tissue of mice which have been
treated with carcinogenic urethane.
Furthermore, the effects of preventing cancer and cancer metastasis
provided by the Minari extract of the present invention can be confirmed
by examining to what extent the oxidative damage to DNA is inhibited
by diets containing Minari extract at various concentrations, based
on measurements of the 8-OH deoxyguanosine level of DNA in the lung
tissue of mice which have been treated with carcinogenic urethane.
Furthermore, the effects of preventing cancer and cancer metastasis
provided by the Minari extract of the present invention can be confirmed
by examining whether or not diets containing Minari extract at various
concentrations obstructs the occurrence of lung cancer in mice which
have been treated with NNK, which is a potential carcinogen in cigarettes.
Furthermore, the effects of preventing cancer and cancer metastasis
provided by the Minari extract of the present invention can be confirmed
by examining to what extent the expression level of PCNA (proliferative
cell nuclear antigen) in the lung tissue of mice which have been
treated with NNK, which is a potential carcinogen in cigarettes,
is suppressed by diets containing Minari extract at various concentrations.
Hereinafter, the present invention will be described by way of
examples.
EXAMPLE 1
Effective Inhibition of Urethane Activity by Minari Extract
(Step 1: Lung Cancer Prevention)
A/J mice were divided in three groups. Through a single gavage,
urethane was administered to each group at 10,000, 20,000, and 40,000
times (125, 250, and 500 mg/kg b.w.) the human urethane exposure
level.
Next, each group was further divided into two experimental groups:
one of the groups (Minari-treated group) was allowed to take, ad
lib., a feed to which dried Minari extract was added to 5% based
on the feed for 4 months, while the other group (urethane control
group) received a normal feed. The results are shown in FIG. 1.
The incidence of lung cancer is significantly reduced in the Minari-treated
groups (p<0.05).
(Step 2: Suppression of the Expression Level of the Cyclin D1 Oncogenic
Gene)
A/J mice were divided into a control group and a urethane-receiving
group. The urethane-receiving group was further divided into three
groups, which respectively received urethane at doses of 125, 250,
or 500 mg/kg. Each urethane-receiving group was further divided
into two experimental groups: one of the groups (urethane-only-receiving
group) received a normal feed, while the other group (urethane+Minari
group) received a feed to which Minari extract was added at a 5%
concentration for 4 months. The expression level of cyclin D1 was
measured after completion of the experiment (The 91st Cancer Society,
presentation article No. 5313: I. P. Lee, Pulmonary cyclin D1-induction
by Fumonisin B1 in Female A/J mice, Apr. 15, 2000). The results
are shown in FIG. 2. The urethane-only-receiving groups showed four
times as high results as those of the Minari-treated groups.
(Step 3: Inhibition of PCNA Expression)
A/J mice were divided into a control group and a urethane-receiving
group. The urethane-receiving group was further divided into three
groups, which respectively received urethane at doses of 125, 250,
or 500 mg/kg. Each urethane-receiving group was further divided
into two experimental groups: one of the groups (urethane-only-receiving
group) received a normal feed, while the other group (urethane+Minari
group) received a feed to which Minari extract was added at a 5%
concentration for 4 months. The PCNA (proliferative cell nuclear
antigen) in the S-phase of the cell cycle was measured after completion
of the experiment (The 91st Cancer Society, presentation article
No. 5313: I. P. Lee, Pulmonary cyclin D1-induction by Fumonisin
B1 in Female A/J mice, Apr. 15, 2000). The results are shown in
FIG. 3. The PCNA level of the urethane-only-receiving groups had
been increased to be four times as high as those of the urethane+Minari
groups.
(Step 4: Suppression of DNA Damage as Indicated by 8-OH Deoxyguanosine
Concentration)
A/J mice were divided into a control group and a urethane-receiving
group. The urethane-receiving group was further divided into three
groups, which respectively received urethane at doses of 125, 250,
or 500 mg/kg. Each urethane-receiving group was further divided
into two experimental groups: one of the groups (urethane-only-receiving
group) received a normal feed, while the other group (urethane+Minari
group) received a feed to which Minari extract was added at a 5%
concentration for 4 months. The 8-OH deoxyguanosine concentration
was measured after completion of the experiment. The results are
shown in FIG. 4. The 8-OH deoxyguanosine concentration in the Minari-treated
groups had been significantly reduced relative to those of the urethane-only-treated
groups.
EXAMPLE 2
Minari Extract which Effectively Suppresses NNK Activation and
Prevents Cancer
(Step 1: Obstruction of Occurrence of Lung Cancer)
NNK, a potential carcinogen in cigarette smoke, was administered
to a total of four groups of A/J mice: an NNK-only-receiving (+normal
feed) group; and NNK+Minari-added-feed-receiving groups (1.25, 2.5
or 5% (w/w)) Such a dose of NNK corresponds to an amount equivalent
to smoking a package of cigarettes for 100 years or more (Djordijevic,
M. V. et al., Comparison of selected components in the mainstream
smoke of the leading U.S. and Japanese cigarettes. In: Proceedings
of the CORESTA smoke and Technology Meeting, pp. 200-217, Nov. 3-8,
1996). Four months later, the lung cancer multiplicity in the A/J
mice was measured.
The results indicate, as shown in FIG. 5, a significantly lower
cancer incidence in the groups receiving pulverized Minari extract
at 2.5% or 5% (p<0.05).
(Step 2: Suppression of the Cyclin D1 Oncogene)
A/J mice were divided into three experimental groups: a single
intraperitoneal water administration-receiving (+normal feed) group;
a single intraperitoneal NNK (4 mg) administration-receiving (+normal
feed) group; and a single intraperitoneal NNK administration-receiving
(+feed to which Minari extract is added at a concentration of 1.25%)
group. Four months later, the expression level of the cyclin D1
oncogene in each experimental group was measured. The results indicate,
as shown in FIG. 6, that the expression level of cyclin D1, an oncogene,
had been reduced to a half or less of that of an NNK control group
(p<0.05). From this, it was indicated that the Minari extract
significantly inhibits NNK activity with respect to the expression
of cyclin D1.
(Step 3: Inhibition of PCNA Expression)
A/J mice were divided into three experimental groups: a single
intraperitoneal water administration-receiving (+normal feed) group;
a single intraperitoneal NNK (4 mg) administration-receiving (+normal
feed) group; and a single intraperitoneal NNK administration-receiving
(+feed to which Minari extract is added at a concentration of 1.25%)
group. Four months later, the expression level of PCNA in each experimental
group was measured. The results indicate, as shown in FIG. 7, that
the expression level of PCNA had significantly increased in the
NNK-treated group, as compared to the water-only-receiving group.
On the other hand, the PCNA expression level was significantly inhibited
in the group which received Minari treatment (1.25% in the feed,
w/w) in addition to NNK administration , as compared to the water-receiving
group (p<0.05).
Industrial Applicability
The inventor has confirmed that Minari extract can obstruct lung
cancer induced by urethane which is present in alcoholic beverages
and carcinogenic foods or by NNK[{(4-N-methyl-N-nitrosoamino)-1-(3-pyridyl)-1-buta
none}] present in cigarette smoke, thereby accomplishing the present
invention. The possible mechanism of cancer prevention by Minari
extract is presumably via obstruction of the activation of precarcinogens
into more active carcinogens, and via removal of free radicals,
which are present in cigarette smoke and alcoholic beverages, by
a number of free radical scavengers within Minari extract. Furthermore,
an objective of the present invention is to prevent, typically,
the occurrence of lung cancer and consequent cancer metastasis due
to carcinogens targeting the lungs, by the use of a dietary ingredient
of Minari extract containing the very effective chemical preventive
substance.
As specifically described in the examples, Minari extract potentially
prevents lung cancer or other cancer induced by carcinogens present
in fermented foods, alcoholic beverages, or cigarette smoke, and
is very important to the medical industry.
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