U.S. patent application number 16/466353 was filed with the patent office on 2020-02-27 for composition for preventing or treating gastritis or peptic ulcer.
The applicant listed for this patent is CHONG KUN DANG PHARMACEUTICAL CORP.. Invention is credited to Jong Lae LIM, Jaehoon YOU.
Application Number | 20200061142 16/466353 |
Document ID | / |
Family ID | 62840462 |
Filed Date | 2020-02-27 |
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United States Patent
Application |
20200061142 |
Kind Code |
A1 |
YOU; Jaehoon ; et
al. |
February 27, 2020 |
COMPOSITION FOR PREVENTING OR TREATING GASTRITIS OR PEPTIC
ULCER
Abstract
The present invention relates to a pharmaceutical or food
composition comprising Cinnamomum cassia pretreated extract
exhibiting an excellently enhanced pharmacological effect than
Cinnamomum cassia extract prepared by a conventional common
extraction method of Cinnamomum cassia and extract prepared with
Cinnamomum cassia single herb medicine, or a method for preventing,
improving or treating gastritis or peptic ulcer using the
composition. In addition, the present invention relates to a
preparation method of Cinnamomum cassia pretreated extract which is
characterized by pretreating Cinnamomum cassia with a non-polar
solvent and extracting it with a polar solvent.
Inventors: |
YOU; Jaehoon; (Yongin-si,
KR) ; LIM; Jong Lae; (Yongin-si, KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
CHONG KUN DANG PHARMACEUTICAL CORP. |
Seoul |
|
KR |
|
|
Family ID: |
62840462 |
Appl. No.: |
16/466353 |
Filed: |
November 2, 2017 |
PCT Filed: |
November 2, 2017 |
PCT NO: |
PCT/KR2017/012349 |
371 Date: |
June 4, 2019 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 1/04 20180101; A61K
36/54 20130101; A23L 33/105 20160801 |
International
Class: |
A61K 36/54 20060101
A61K036/54; A23L 33/105 20060101 A23L033/105 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 11, 2017 |
KR |
10-2017-0004418 |
Claims
1. A pharmaceutical composition for preventing or treating
gastritis or peptic ulcer comprising non-polar solvent pretreatment
and polar solvent extract of Cinnamomum cassia.
2. The pharmaceutical composition according to claim 1, wherein the
peptic ulcer is stomach ulcer or duodenal ulcer.
3. The pharmaceutical composition according to claim 1, wherein the
non-polar solvent is ethyl acetate.
4. The pharmaceutical composition according to claim 1, wherein the
polar solvent is water.
5. The pharmaceutical composition according to claim 1, which is
administered once or several times as divided, wherein the
pharmaceutical composition is characterized by comprising the
extract so that the daily dose of the Cinnamomum cassia pretreated
extract is 9.7 to 2,919 mg.
6. The pharmaceutical composition according to claim 1, wherein the
yield of the Cinnamomum cassia pretreated extract is 15 to
20.fwdarw.1 (meaning that 1 Kg of the final extraction dry ex is
obtained when adding 15 to 20 Kg of crude herb medicine).
7. A food composition for preventing or improving gastritis or
peptic ulcer comprising non-polar solvent pretreatment and polar
solvent extract of Cinnamomum cassia.
8. The food composition according to claim 7, wherein the peptic
ulcer is stomach ulcer or duodenal ulcer.
9. The food composition according to claim 7, wherein the non-polar
solvent is ethyl acetate.
10. The food composition according to claim 7, wherein the polar
solvent is water.
11. The food composition according to claim 7, which is
administered once or several times as divided, wherein the
Cinnamomum cassia pretreated extract is characterized by comprising
the extract so that the daily dose of the Cinnamomum cassia
pretreated extract is 9.7 to 2,919 mg.
12. The food composition according to claim 1, wherein the
composition is health functional food, health supplementary food or
functional food.
13. A preparation method of Cinnamomum cassia non-polar solvent
pretreatment and polar solvent extract comprising a) a step of
pretreating Cinnamomum cassia by immersing into a non-polar
solvent; b) a step of removing the non-polar solvent in the step
a); and c) a step of extracting Cinnamomum cassia obtained in the
step b) with a polar solvent.
14. The preparation method according to claim 13, wherein the
non-polar solvent in the step a) is ethyl acetate.
15. The preparation method according to claim 13, wherein the step
a) is performed by immersing Cinnamomum cassia into a non-polar
solvent having a weight of 1 to 3 times compared to Cinnamomum
cassia herb medicine and stirring at 20 to 35.degree. C. for 30
minutes to 3 hours.
16. The preparation method according to claim 13, wherein the polar
solvent used in the step c) is water.
17. The preparation method according to claim 13, wherein the step
c) is performed by extracting Cinnamomum cassia obtained in the
step b) for 80 to 100.degree. C. for 2 hours to 6 hours using a
polar solvent of 6 times to 10 times compared to Cinnamomum cassia
herb medicine.
18. The preparation method according to claim 13, wherein the dry
yield of the extract prepared in the step c) is
15.about.20.fwdarw.1 (meaning that 1 Kg of the final extraction dry
ex is obtained when adding 15 to 20 Kg of crude herb medicine).
19. A method for preventing, improving or treating gastritis or
peptic ulcer, comprising a step of administering a composition
comprising non-polar solvent pretreatment and polar solvent extract
of Cinnamomum cassia.
Description
TECHNICAL FIELD
[0001] The present invention relates to an enhanced composition
capable of preventing or treating gastritis or peptic ulcer such as
stomach ulcer. More specifically, the present invention relates to
a composition for treating gastritis or stomach ulcer comprising
Cinnamomum cassia extract, showing an excellently enhanced
pharmacological effect than Cinnamomum cassia extract prepared by a
conventional common extraction method of Cinnamomum cassia and
extract prepared with Cinnamomum cassia single herb medicine, and a
preparation method thereof.
BACKGROUND ART
[0002] Stomach is a part of digestive tract, and is a part inflated
as a pocket between esophagus and small intestine (duodenum), and
it is an organ which stores food entered through esophagus and
breaks down it to facilitate digestion, and controls to send food
to duodenum and is harmonized with secretion of digestive enzymes,
thereby allowing efficient digestion and absorption. Factors that
adversely affect a person's gastrointestinal function are extremely
diverse in nature and can occur in the upper gastrointestinal
tract, the lower gastrointestinal tract or both, and include a wide
range of gastrointestinal disorders including genetic,
physiological, environmental and mental factors. The representative
diseases of the upper gastrointestinal tract are gastritis and
peptic ulcer including stomach ulcer and duodenal ulcer. Gastritis
presents damage and inflammation of stomach mucosa, and stomach
ulcer means when such damage penetrates the mucosa and invades
submucosa tissue and muscle layer. In addition, duodenal ulcer is
ulcer occurred in duodenum and is also called peptic ulcer by
collectively calling stomach ulcer and duodenal ulcer. These
gastritis and peptic ulcer have been known to be occurred by
imbalance of gastric acid, anti-inflammatory drug and bacterial
inflammation called offensive factors, and mucus, cell
regeneration, alkali secretion and so on called defensive
factors.
[0003] Antacids which neutralize excess secreted gastric acid,
histamine antagonists for inhibition of acid secretion, proton pump
inhibitors, anticholinergics, stomach mucosa protectants increasing
resistance of stomach intimae to the digestive solution and helping
recovery, and the like are mainstreams for treatments of gastritis
and peptic ulcer, and recently, there has been a drug treatment for
prescribing the above drug and antibiotics in combination to remove
Helicobacter pylori. The characteristics of antacids are rapid
acting, and defensing damage of stomach mucosa by gastric acid by
neutralizing gastric acid by raising pH in stomach. However, it
affects the gastrointestinal tract smooth muscles by administration
of inorganic materials, and thereby it may cause constipation or
diarrhea, or cause allergic rejection.
[0004] Cimetidine is a representative of inhibitors of histamine
receptor blocking secretion widely used for peptic ulcer, and there
are ranitidine, famotidine, roxatidine, and the like as its
derivatives, but they block histamine receptors in stomach mucosa
and therefore histamine molecules act to prevent stomach cells from
secreting acid. It shows an excellent anti-ulcer effect in clinical
practice, but there is a disadvantage of high recurrence rate,
since it is easily damaged by offensive factors such as gastric
acid and the like after drug administration cessation, as cured
regenerated mucosa and submucosa tissue have a week structure
compared to normal tissue. In addition, ranitidine also has a
disadvantage in that it is not effective for diseases such as acute
gastritis caused by ethanol, and therefore the stomach mucosa
protecting ability is not good.
[0005] As relatively recently developed proton pump inhibitors,
there are omeprazole and lansoprazole and the like, and they are
known to exhibit a strong acid secretion inhibitory effect by
inhibiting acid secretion at the final stage from gastric wall
cells, but it has a high recurrence rate and side effects such as
diarrhea, fever, headache, fatigue and so on have been
reported.
[0006] Stomach mucosa protectants are known to have a disadvantage
in that it generally requires a long-term treatment and has a large
dosage, but regenerated mucosa is recovered similarly to the normal
different from offensive factor inhibitors.
[0007] Helicobacter pylori that is a type of bacteria inhabiting
stomach is known as a representative cause of recurrence of peptic
ulcer. Helicobacter pylori is a gram-negative bacillus that
inhabits stomach mucosa epithelial intercellular junctions and
causes chronic stomach ulcer, and treatment has been carried out to
eradicate it and many achievements have been made to date, but
there are problems such as efficacy, side effects and emergence of
resistant strains and the like, and safe and reliable eradication
methods have not yet been established.
[0008] On the other hand, Cinnamomum cassia is a Ranunculales
Lauraceae evergreen broad-leaved tree belonging to a dicotyledon
and it is originated from China, and is distributed in Sri Lanka,
Indochina and Korea (Jeju), and it is a medicinal use name of
branches and bark of Cinnamomum cassia tree (Cinnamon tree) that
grows up to about 8 m in height in mountain.
[0009] Regarding components or physiological activities of such
Cinnamomum cassia, researches on antidiabetic effect (Boaduo N K et
al., Pharm Biol. 2014, 52(6):756-61; Han Y et al., Pharm Biol.
2013, 51(8):961-7), infection prevention effect (Yeh C F et al., J
Ethnopharmacol. 2013, 147(2):321-6), antioxidant action (Hwa J S et
al., J Ethnopharmacol. 2012, 139(2):605-15), anti-stomach ulcer
action of eugenol and cinnamic acid (Jung J et al., Yakugaku
Zasshi. 2011, 131(7):1103-10), antifibrotic action (Lim C S et al.,
Biosci Biotechnol Biochem. 2010, 74(3):477-83), melanin synthesis
inhibitory action (Kong Y H et al., Biol Pharm Bull. 2008,
31(5):946-8), osteogenesis promoting action (Lee K H et al.,
Phytother Res. 2006, 20(11):952-60), antibiosis (Ooi L S et al., Am
J Chin Med. 2006, 34(3):511-22) and so on were known.
[0010] In this regard, Korean Patent Publication No.
10-2003-0030422 discloses a health supplementary food for improving
stomach functions comprising the extract obtained by hot water
extraction of Alpiniae semen, Amomi semen, Cinnamomum cassia,
Astragalus membranaceus and Citrus unshiu peel at the ratio of
1:1:1:1 as an active ingredient.
[0011] On the other hand, cinnamic aldehyde which is the main
component and the active ingredient of Cinnamomum cassia is easily
oxidized and colored in air and therefore benzoic acid crystals may
be educed, and thus it has been reported that the stability at the
room temperature is low (Food Additives Codex, JongHee P et al,
2006), and 2-methoxycinnamic aldehyde has been also reported that
the stability at the room temperature is low (Jeongsook L et al,
2013). Although these components of Cinnamomum cassia are known as
index components, but it has been reported that there is risk that
the stability of drugs are considerably degraded by these
substances when preparing drugs from the Cinnamomum cassia extract
comprising them.
[0012] Thus, the development of a preparation for preventing and
treating gastritis and stomach ulcer which minimizes the ingredient
content that may affect the stability and at the same time maximize
the effectiveness has been still strongly required.
[0013] Lastly, it may be an important factor to reduce the dosage
of the extract in a pharmaceutical composition. As reducing
formulations such as tablets, syrup, and the like of drugs is
considerably affected by the method for preparing the extract, it
can be a core factor to reduce the size and volume of preparations
in a lower volume than conventional common extract, and at the same
time obtain the same effect, by selectively extracting the
effective part.
[0014] Under these circumstances, the present inventors have tried
to develop Cinnamomum cassia extract which has anti-ulcer efficacy
through much excellent defensive factor increasing mechanisms
compared to Artemisia extract preparation that is a commercially
available herb medicine-based therapeutic agent for gastritis and
stomach ulcer and shows much excellent pharmacological effects than
the Cinnamomum cassia extract obtained by water extraction, alcohol
extraction, solvent fractionation, and so on, or the extract
prepared by Cinnamomum cassia single herb medicine. As a result, it
was confirmed that when preparing extract by extracting with a
specific solvent after pretreatment of a specific solvent to
Cinnamomum cassia different from extraction, it exhibits an
excellently enhanced gastritis or stomach ulcer preventing or
treating effect than conventional extract and therefore not only it
can effectively improve gastritis and peptic ulcer such as stomach
ulcer, but also it can minimize components known to affect the
stability such as cinnamic aldehyde and 2-methoxycinnamic aldehyde
and the like and at the same time provide a preparation with very
improved effectiveness, thereby completing the present
invention.
DISCLOSURE
Technical Problem
[0015] An object of the present invention is to provide a
composition comprising Cinnamomum cassia pretreated extract with an
enhanced effect of preventing, improving or treating gastritis or
stomach ulcer, a method for preventing, improving or treating
gastritis or stomach ulcer using thereof, and a preparation method
of the Cinnamomum cassia pretreated extract.
[0016] More specifically, one object of the present invention is to
provide a pharmaceutical or food composition for preventing or
treating gastritis or stomach ulcer comprising Cinnamomum cassia
pretreated extract which is obtained by pretreating Cinnamomum
cassia with a non-polar solvent and then extracting it with a polar
solvent, a method for preventing, improving or treating gastritis
or stomach ulcer using thereof, and a preparation method of the
Cinnamomum cassia pretreated extract.
Technical Solution
[0017] One aspect to achieve the object, the present invention
relates to a composition for preventing, improving or treating
gastritis or peptic ulcer comprising Cinnamomum cassia pretreated
extract which is obtained by pretreating Cinnamomum cassia with a
non-polar solvent and extracting with a polar solvent, and a method
for preventing, improving or treating gastritis or stomach ulcer
using thereof.
[0018] The composition of the present invention may be a
pharmaceutical composition or a food composition.
Advantageous Effects
[0019] The composition of the present invention comprises
Cinnamomum cassia pretreated extract prepared through a unique
process as an active ingredient, thereby not only showing excellent
effects of stomach ulcer inhibition, mucosal amount improvement and
inhibition of inflammation, but also minimizing cinnamic aldehyde
and 2-methoxycinnamic aldehyde which may affect stability, to
minimize stability hindrance factors, and it can provide a
preparation in which problems such as toxicity, and so on are
solved, as it is based on herb medicine, and therefore it has an
excellent effect compared to conventional herb medicine-based
drugs, and the size or dose of a formulation can be reduced due to
properties showing the same effect with less dose, and thus it is
useful for development of a preparation for prevention, improvement
and treatment of gastritis or peptic ulcer.
BRIEF DESCRIPTION OF DRAWINGS
[0020] FIG. 1 is a photograph confirming the effect of inhibiting
stomach damage of the composition according to the present
invention, and
[0021] FIG. 2 is a graph showing the effect of inhibiting stomach
damage of the composition according to the present invention with a
stomach ulcer index.
BEST MODE
[0022] Hereinafter, the present invention will be described in more
detail.
[0023] The composition according to the present invention is
characterized by comprising extract which is obtained by
pretreating Cinnamomum cassia with a non-polar solvent and then
extracting with a polar solvent as an active ingredient. Cinnamomum
cassia used as crude herb medicine for the pretreated extract of
the composition is a Ranunculales Lauraceae evergreen broad-leaved
tree belonging to a dicotyledon and it is originated from China,
and is distributed in Sri Lanka, Indochina and Korea (Jeju), and it
means branches and bark of Cinnamomum cassia tree (Cinnamon tree)
that grows up to about 8 m in height in mountain.
[0024] In the composition according to the present invention, the
Cinnamomum cassia may be used as dried or cut in an appropriate
size for extraction according to conventional known methods, and so
on. In addition, the present invention is pretreated extract before
extracting such Cinnamomum cassia crude herb medicine with a polar
solvent, and such pretreatment means treatment with a non-polar
solvent. For example, the non-polar solvent is characterized by
being ethyl acetate, and a non-polar solvent up to 1 time to 3
times of solvent volume compared to the Cinnamomum cassia herb
medicine weight may be used. As one preferable aspect, pretreatment
may be performed by immersing and stirring sliced Cinnamomum cassia
in the non-polar solvent like ethyl acetate for about 30 minutes to
3 hours at a range of 20 to 35.degree. C., preferably, at a room
temperature. During this pretreatment process, cinnamic aldehyde
and 2-methoxycinnamic aldehyde that may adversely affect the
stability may be effectively removed. Furthermore, the pretreated
extract according to the present invention is characterized as
polar solvent, preferably, water extract of Cinnamomum cassia
pretreated as above. Extraction may be carried out at 80 to
100.degree. C. for 2 hours to 6 hours, preferably 5 hours, by using
about 6 times to 10 times volume, preferably 8 times volume of the
polar solvent used for extraction compared to the Cinnamomum cassia
herb medicine weight. This extraction may be performed once to
several times, and the extracted pretreated extract may be further
filtered, concentrated and dried, and methods used then may use
filtering, concentration and drying methods commonly used for
preparation of extract without limitations.
[0025] In one embodiment according to the present invention,
Cinnamomum cassia was dried and sliced and was immersed and stirred
at a room temperature over one hour by adding 2 times of ethyl
acetate to Cinnamomum cassia, and after removing ethyl acetate,
with water, pretreated Cinnamomum cassia herb medicine was washed,
and water corresponding to 8 times of Cinnamomum cassia herb
medicine was added and it was extracted at about 90.degree. C. for
5 hours, and this was repeated twice. Accordingly, the collected
extract was filtered, vacuum concentrated, vacuum dried or spray
dried, to prepare Cinnamomum cassia ethyl acetate pretreated water
extract.
[0026] The pharmaceutical composition of the present invention
exhibits an excellent effect for preventing or treating gastritis
or peptic ulcer like stomach ulcer and duodenum ulcer.
[0027] The pharmaceutical composition of the present invention may
comprise the Cinnamomum cassia pretreated extract of 10 to 90% by
weight based on the total weight of composition. This may be
increased or decreased according to needs of users, and it may be
increased or decreased appropriately according to circumstances
such as dietary life, nutritive conditions, stomach inflammation or
ulcer degree, progression of the disease, and the like.
[0028] The pharmaceutical composition may be administered orally or
parenterally, and it may be used in a form of general drug
preparations. Preferable pharmaceutical preparations include
preparations for oral administration such as tablets, pills,
powders, granules, hard or soft capsules, liquids, suspensions, and
the like, and these pharmaceutical preparations may be prepared
using a pharmaceutically acceptable common carrier, for example, in
case of preparations for oral administration, excipients, binding
agents, disintegrating agents, lubricants, solubilizers, suspending
agents, preservatives or extenders and so on.
[0029] The dosage of the pharmaceutical composition of the present
invention may be determined by an expert according to various
factors such as conditions, age, body weight of patients, disease
progression and the like, but generally 9.7.about.2,919 mg of
extract a day may be administered once to several times as divided,
and preferably, 29.2.about.2,919 mg, more preferably
68.1.about.2,919 mg may be administered. However, in case of a
long-term intake, the amount may be the above range or less, and
since the active ingredient has no problem in terms of stability,
the amount over the above range may be used.
[0030] In addition, the composition of the present invention may be
a food composition. The food may be health supplementary food,
health functional food, functional food, and so on, but not limited
thereto, and it includes natural food, processed food, general food
materials and the like in which the Cinnamomum cassia pretreated
extract of the present invention is added. Herein functionality
means obtaining a useful effect for health care uses like nutrient
control or physiological actions for the structure and function of
a human body. The health supplementary food, health functional food
or functional food of the present invention may be prepared by
methods commonly used in the art, and during the preparation, it
may be prepared by adding raw materials and components commonly
added in the art. Moreover, different from general drugs, it has an
advantage of no side effects which may be caused in case of
long-term use of drugs, since it uses herb medicine fed currently
as raw materials, and it has outstanding portability, and therefore
the health functional food of the present invention can be ingested
as an adjuvant for increasing the effect of preventing or improving
gastritis or stomach ulcer. The mixed amount of active ingredients
may be suitably determined according to use purposes (prevention,
health or therapeutic treatment). Generally, 10 to 90% by weight of
the Cinnamomum cassia pretreated extract according to the present
invention may be comprised during food preparation. In case of
long-term ingestion on purpose of health control, the amount may be
used in the above range or less.
[0031] The effective dose may be used following the effective dose
of the pharmaceutical composition, but in case of long-term
ingestion for improvement or maintenance of gastritis or stomach
ulcer, it may be the above range or less, and the effective
ingredients may be used over the above range, as they are herb
medicine components and have no problem in terms of stability.
[0032] There is no particular limitation on the kind of the food.
The food composition comprising the Cinnamomum cassia pretreated
extract may be used in a form of preparations for oral
administration such as tablets, hard or soft capsules, liquids,
suspensions, and the like, and these preparations may further
comprise an acceptable food supplementary additive. The term "food
supplementary additive" is added for preparing each formulation of
health functional food, and it may be selected appropriately by
those skilled in the art can select and used. Examples of the food
supplementary additive include various nutritional supplements,
vitamins, minerals (electrolytes), flavors such as synthetic
flavors and natural flavors and the like, coloring agents and
fillers, pectic acid and its salt, alginic acid and its salt,
organic acids, protective colloid thickeners, pH adjusting agents,
stabilizers, preservatives, glycerin, alcohol, carbonating agents
used for carbonated beverages, and so on, but the kind of the food
supplementary additive of the present invention is not limited by
these examples.
[0033] In addition, the food composition comprising the Cinnamomum
cassia pretreated extract of the present invention may be added by
itself or used with other food or food composition, and may be
appropriately used according to common methods. The mixed amount of
effective ingredients may be suitably determined according to its
use purposes (prevention, improvement or therapeutic
treatment).
[0034] Examples of the food in which the Cinnamomum cassia
pretreated extract may be added are meat, sausage, bread,
chocolate, candies, snack, confectionery, pizza, ramen, other
noodles, gum, dairy products including ice cream, various kinds of
soup, beverages, tea, drinks, alcohol beverages and vitamin
complex, other nutritional supplements, and the like, but is not
limited by these kinds of food.
[0035] Furthermore, as another aspect, the present invention
relates to a preparation method of the Cinnamomum cassia pretreated
extract according to the present invention comprising the following
steps:
[0036] a) a step of pretreating Cinnamomum cassia by immersing it
in a non-polar solvent;
[0037] b) a step of removing the non-polar solvent in the step a);
and
[0038] c) a step of extracting the Cinnamomum cassia collected in
the step b) with a polar solvent.
[0039] The non-polar solvent used in the step a) of the preparation
method is characterized by being ethyl acetate, and a non-polar
solvent up to 1 time to 3 times of solvent volume compared to the
Cinnamomum cassia herb medicine may be used. As one preferable
aspect, pretreatment may be performed by immersing and stirring
sliced Cinnamomum cassia in the non-polar solvent like ethyl
acetate for about 30 minutes to 3 hours at a range of 20 to
35.degree. C., preferably, at a room temperature. During this
pretreatment process, cinnamic aldehyde and 2-methoxycinnamic
aldehyde that may adversely affect the stability may be effectively
removed.
[0040] The step b) is a step of removing the non-polar solvent from
the Cinnamomum cassia pretreated with the non-polar solvent and
known methods for removing a non-polar solvent may be used without
limitations.
[0041] The step c) may be performed by extracting at 80 to
100.degree. C. for 2 hours to 6 hours, preferably 5 hours, using
about 6 times to 10 times, preferably 8 times of weight compared to
Cinnamomum cassia herb medicine. Such a step c) may be performed
once to several times.
[0042] As an additional aspect, the pretreated extract prepared in
the step c) may be filtered, concentrated and dried, and for
methods used then, filtering, concentration and drying methods
commonly used for preparation of extract may be used without
limitations. In one specific aspect, when preparing the Cinnamomum
cassia pretreated extract by the preparation method according to
the present invention, the yield is 15 to 20.fwdarw.1 (meaning that
1 Kg of the final extraction dry ex is obtained when adding 15 to
20 Kg of crude herb medicine).
[0043] According to one example of the present invention, the
Cinnamomum cassia pretreated extract prepared according to the
preparation method of the present invention not only exhibits
excellent effects of stomach ulcer inhibition, stomach mucosal
amount improvement and inhibition of inflammation, but also
exhibits an enhanced effect of minimizing cinnamic aldehyde and
2-methoxycinnamic aldehyde which may affect stability, to minimize
stability hindrance factors, compared to a conventional herb
medicine-based therapeutic agent for gastritis and stomach ulcer,
Artemisia extract preparation, as well as simple water extract of
Cinnamomum cassia and extract in which Cinnamomum cassia is
pretreated with other solvent.
MODE FOR INVENTION
[0044] Hereinafter, the present invention will be described in
detail by examples. However, the following examples are intended to
illustrate the present invention only, and the present invention is
not limited by the following examples.
Example 1. Preparation of Cinnamomum cassia Pretreated Extract
According to the Present Invention
[0045] The Cinnamomum cassia pretreated extract according to the
present invention was prepared as follows. Specifically, 2 times of
ethyl acetate was added to Cinnamomum cassia herb medicine and it
was immersed and stirred at a room temperature for over 1 hour.
After removing ethyl acetate and washing Cinnamomum cassia herb
medicine with water, 8 times of water was added and it was
extracted at near about 90.degree. C. for 5 hours (2 repeats). The
Cinnamomum cassia pretreated water extract was prepared by
filtering, vacuum concentrating, vacuum drying or spray drying the
extract (extract yield: 15.about.20.fwdarw.1).
Comparative Examples 1 to 6. Preparation of Cinnamomum cassia
Solvent Extract, Cinnamomum cassia Pretreated Extract Using Other
Solvent and Commercial Preparations
[0046] For comparison with the Cinnamomum cassia pretreated extract
according to the present invention, Cinnamomum cassia solvent
extract and Cinnamomum cassia butanol pretreated extract, and
Ranitidine, Rebamipide and Artemisia extract as commercial
preparations were prepared.
[0047] 1) Cinnamomum cassia Ethanol Extract
[0048] To Cinnamomum cassia herb medicine, 8 times of ethanol was
added and it was extracted at near about 90.degree. C. for 5 times
(2 repeats). The extract was filtered, vacuum concentrated, vacuum
dried or spray dried to use it as Cinnamomum cassia ethanol extract
(extract yield: 25.fwdarw.1).
[0049] 2) Cinnamomum cassia Water Extract
[0050] The extract was prepared by the same method with the
Cinnamomum cassia ethanol extract, but only the extraction solvent
was changed into water (extract yield: 11.about.13.fwdarw.1).
[0051] 3) Cinnamomum cassia Butanol Pretreated Water Extract
[0052] The extract was prepared by the same method with the Example
1, but only the pretreatment solvent was changed into butanol.
[0053] Only the main ingredients of commercial preparations of
Ranitidine, Rebamipide, and Artemisia extract were used.
TABLE-US-00001 TABLE 1 Kinds of comparative preparations
Comparative example Ingredient name Comparative example 1
Cinnamomum cassia ethanol extract Comparative example 2 Cinnamomum
cassia water extract Comparative example 3 Cinnamomum cassia
butanol pretreated water extract Comparative example 4 Ranitidine
Comparative example 5 Rebamipide Comparative example 6 Artemisia
extract
Experimental Example 1. Confirmation of Inhibition Rate of Stomach
Ulcer
[0054] In order to confirm the inhibition rate of stomach ulcer of
the Example 1 and Comparative examples 1 to 6, the test was carried
out according to the following method.
[0055] Test animals were randomly assigned to 10 animals of
specific pathogen free (SPF) 7-week-old rat males per group so that
the average body weight was maximally uniformly distributed
throughout the adaptation period (7 days). The administration route
was single oral administration of substances of the Example 1 and
Comparative examples 1 to 6, respectively, and the dose was
calculated to be 10 mL/Kg on the basis of the weight measured on
the day of administration. All animals were fasted 48 hours before
administration of test substances and control drug, and then each
of test substances and control drug was orally administered, and
Indomethacin previously prepared 30 minutes after administration
was orally administered at a dose of 80 mg/kg. After anesthetizing
with diethyl ether 5 hours after administration of Indomethacin,
stomach was removed and the gastric mucosa was photographed with a
digital camera. The area of the damaged site was analyzed using
ImageJ software (NIH, Bethesda, Md.). The stomach ulcer index was
measured according to the following Equation 1. The result was
shown in the following Table 2, FIG. 1 and FIG. 2.
Stomach ulcer index (%)=(damage area/total area).times.100
[Equation 1]
TABLE-US-00002 TABLE 2 Inhibition rate of stomach ulcer compared to
the induced group Inhibition rate (%), Stomach ulcer compared to
the Group Dose(mg/kg) index (%) induced group Induced group 0 6.58
.+-. 1.60 -- Comparative 150.0 2.00 .+-. 1.05* 69.6 example 2
Comparative 150.0 2.88 .+-. 2.29* 56.2 example 3 Comparative 75.0
0.20 .+-. 0.29* 97.0 example 4 Comparative 150.0 2.35 .+-. 1.83*
64.3 example 6 Example 1 150.0 0.24 .+-. 0.18* 96.4 *showing
significant difference compared to the induced group (p <
0.05)
[0056] As could be seen in Table 2, FIG. 1 and FIG. 2, the
Cinnamomum cassia ethyl acetate pretreated water extract of Example
1 according to the present invention exhibited the 27.4 to 40.0%
increased inhibition rate, compared to not only the simple solvent
extract of Cinnamomum cassia (Comparative example 2), but also the
extract which was obtained by pretreating Cinnamomum cassia with
butanol and extracting with water (Comparative example 3).
Moreover, it could be confirmed that this was the equivalent level
of inhibition rate to the strong stomach ulcer inhibitor,
Ranitidine (Comparative example 4). It could be seen that the rat
effective does of Example 1 (150 mg/kg) was 1,460 mg when converted
into the dose per body weight 60 Kg adult unit area.
Experimental Example 2. Confirmation of Effect of Improving Mucosal
Amount
[0057] In order to confirm the effect of improving the mucosal
amount of the Example 1 and Comparative examples 1 to 6, the test
was carried out according to the following method.
[0058] Test animals were randomly assigned to 10 animals of
specific pathogen free (SPF) 7-week-old rat males per group so that
the average body weight was maximally uniformly distributed
throughout the adaptation period (7 days). All animals were fasted
48 hours before administration and then test substances and control
drug were orally administered, and 30 minutes after administration,
Indomethacin 80 mg/kg was orally administered. It was anesthetized
with diethyl ether 7 hours after administration of Indomethacin and
blood was collected from postcaval vein, and then stomach was
removed and was incised along with greater curvature and was turned
over and the glandular ventriculus mucosal surface was exposed
outside and the glandular ventriculus was washed out with 0.25M
sucrose solution. After soaking in 0.1% w/v alcian blue 8 GX
solution 15 mL for 2 hours, it was washed with 15 mL of 0.25 M
sucrose solution twice (15 minutes each) to remove excessive amount
of dye. To extract a reagent combined with stomach mucus, the
stomach tissue was soaked in 15 mL of 0.5 M MgCl.sub.2 solution for
2 hours and shaking was carried out intermittently for 1 minute
each at intervals of 30 minutes. After mixing the extract solution
with diethyl ether in the same amount and shaking, it was
centrifuged at 3,500 rpm for 10 minutes and for the supernatant,
the absorbance was measured at 605 nm. 0.1% w/v alcian blue 8 GX
solution was diluted with 0.25 M sucrose solution and standard
solution was prepared and thereby a standard curve was fabricated
and the measured absorbance value was converted into the amount of
alcian blue combined with the stomach mucus, and the result was
shown in Table 3.
TABLE-US-00003 TABLE 3 Confirmation of the effect of improving
mucosal amount Mucosal % compared to the Group Dose(mg/kg) amount
[.mu.g/L] normal group Normal group -- 11.2 .+-. 2.6 100 Induced
group -- 4.8 .+-. 1.9 42.9 Comparative 300.0 8.0 .+-. 2.1* 72.0
example 2 Comparative 100.0 10.6 .+-. 3.7* 94.6 example 5
Comparative 300.0 8.5 .+-. 2.4* 76.4 example 6 Example 1 300.0 9.1
.+-. 2.7* 81.3 *showing significant difference compared to the
induced group (p < 0.05)
[0059] As could be seen in the Table 3, the Cinnamomum cassia ethyl
acetate pretreated water extract according to the present invention
of Example 1 exhibited 10% increased stomach mucus amount compared
to the simple Cinnamomum cassia water extract (Comparative example
2). In addition, it was confirmed that such an effect of increasing
the stomach mucus amount was increased compared to the commercially
available herb medicine preparation, Artemisia extract (Comparative
example 6). It could be seen that the rat effective does of Example
1 (300 mg/kg) was 2,919 mg when converted into the dose per body
weight 60 Kg adult unit area.
Experimental Example 3. Confirmation of Effect of Improvement of
PGE2
[0060] Since the recovery mechanism of stomach mucous membrane
blood stream and epithelial cells is maintained and the action of
preserving cells of the stomach mucous membrane was shown, when
PGE2 in serum is increased, the enhanced improvement of PGE2
affects treatment of gastritis or stomach ulcer. In order to
confirm the effect of the Example 1 and Comparative examples 2, 5
to 6 according to the present invention for this improvement of
PGE2, for the blood collected in Test example 2, serum was
separated according to the protocol provided by the manufacturer of
PGE2 ELISA Kit and then sampling treatment was carried out, and
then it was analyzed, and the result was shown in the following
Table 4.
TABLE-US-00004 TABLE 4 PGE2 improvement effect % compared to the
Group Dose(mg/kg) PGE2 [ng/mL] normal group Normal group -- 1.88
.+-. 0.2 100 Induced group -- 1.61 .+-. 0.17 0 Comparative 300.0
1.78 .+-. 0.2* 64.4 example 2 Comparative 100.0 1.82 .+-. 0.14*
77.7 example 5 Comparative 300.0 1.73 .+-. 0.39 44 example 6
Example 1 300.0 1.8 .+-. 0.19* 72.6 *showing significant difference
compared to the induced group (p < 0.05)
[0061] As could be confirmed in Table 4, the Cinnamomum cassia
ethyl acetate pretreated water extract according to the present
invention (Example 1) exhibited the very enhanced PGE2 improving
effect compared to the simple Cinnamomum cassia solvent extract
(Comparative example 2), and exhibited the even about 28% increased
improvement rate compared to the commercially available herb
medicine, Artemisia extract (Comparative example 6), and exhibited
the equivalent level of effect to the strong stomach ulcer
therapeutic agent, Rebamipide (Comparative example 5).
Experimental Example 4. Stomach Ulcer Inhibition Rate by Volume
[0062] The stomach ulcer test by volume for the Example 1 was
tested by the method as Experimental example 1, and was shown in
Table 5.
TABLE-US-00005 TABLE 5 Inhibition rate of stomach ulcer by dose
Inhibition rate (%), Stomach ulcer compared to the Group
Dose(mg/kg) index (%) induced group Induced group 0 5.03 .+-. 1.17
0.0 Example 1 1 3.61 .+-. 0.86* 28.1 Example 1 3 2.18 .+-. 1.21*
56.6 Example 1 7 1.46 .+-. 0.70* 70.9 Example 1 15 1.03 .+-. 0.63*
79.5 Example 1 25 0.77 .+-. 0.53* 84.7 Example 1 50 0.48 .+-. 0.29*
90.4 Example 1 100 0.24 .+-. 0.22* 95.3 Example 1 150.0 0.24 .+-.
0.18* 96.4 Comparative 150.0 2.00 .+-. 1.05* 69.6 example 2
*showing significant difference compared to the induced group (p
< 0.05)
[0063] As could be confirmed in Table 5, the Cinnamomum cassia
ethyl acetate pretreated water extract according to the present
invention (Example 1) exhibited the significant stomach ulcer
inhibition effect from 1 mg/kg up to 150 mg/kg to the maximum. It
could be seen that the rat effective does was 1,460 mg to the
maximum at 9.7 mg (converted volume of rat 1 mg/kg), when converted
into the dose per body weight 60 Kg adult unit area.
[0064] In addition, Comparative example 2 (conventional (or common)
Cinnamomum cassia water extract) exhibited the inhibition rate of
69.6% compared to the induced group at 150 mg/kg and it was shown
that the similar effect to the volume of 7 mg/kg of Example 1 of
the present invention was exhibited, and therefore it could be seen
that the same efficacy was exhibited even when reducing about 21
times of volume.
Experimental Example 5. Analysis of Content of Cinnamomum cassia
Pretreated Extract According to the Present Invention
[0065] In order to analyzed the components and contents of the
Cinnamomum cassia pretreated extract according to the present
invention, for the Cinnamomum cassia extract of Example 1, HPLC
content analysis was conducted under the following conditions using
cinnamic acid, cinnamic aldehyde, 2-methoxy cinnamic aldehyde
standard products.
[0066] HPLC Equipment Operating Conditions
[0067] 1) Column: Kromasil 100-5 C.sub.18 (4.6 mm.times.150 mm, 5.0
.mu.m) or column equivalent to the same
[0068] 2) Detector: Ultraviolet spectrophotometer (measurement
wavelength: 280 nm)
[0069] 3) Flow rate: About 0.7 mL/minute
[0070] 4) Dose: 10 .mu.L
[0071] 5) Mobile phase: Acetonitrile (0.2% trifluoroacetic
acid)/purified water
[0072] The result was shown in Table 6.
TABLE-US-00006 TABLE 6 HPLC content analysis 2-methoxy cinnamic
Cinnamic acid Cinnamic aldehyde aldehyde Comparative 2.91 22.3 3.53
example 1 Comparative 5.61 0.3 0.44 example 2 Example 1 4.26 0.07
0.13 [Unit: mg/g]
[0073] Cinnamic aldehyde which is the main component and the active
ingredient of Cinnamomum cassia is easily oxidized and colored in
air and therefore benzoic acid crystals may be educed, and thus it
has been reported that the stability at the room temperature is low
(Food Additives Codex, JongHee P et al, 2006), and
2-methoxycinnamic aldehyde has been also reported that the
stability at the room temperature is low (Jeongsook L et al, 2013).
However, as could be confirmed in Table 5, it was confirmed that
the Cinnamomum cassia pretreated extract according to the present
invention comprised very small amount of the cinnamic aldehyde and
2-methoxycinnamic aldehyde compared to the Cinnamomum cassia
ethanol, Cinnamomum cassia water extract that Cinnamomum cassia was
extracted simply with a solvent. Cinnamic aldehyde was reduced
about 323 times for Comparative example 1 and about 4.3 times for
Comparative example 2, and 2-methoxycinnamic aldehyde was reduced
about 27 times for Comparative example 1 and about 3.4 times for
Comparative example 2. Thus, the Cinnamomum cassia pretreated
extract according to the present invention minimized components
capable of affecting the stability.
* * * * *