U.S. patent application number 14/865845 was filed with the patent office on 2016-01-14 for adrenomedullin production enhancer.
This patent application is currently assigned to National University Corporation ASAHIKAWA MEDICAL COLLEGE. The applicant listed for this patent is National University Corporation ASAHIKAWA MEDICAL COLLEGE, TSUMURA & Co.. Invention is credited to Atsushi KANEKO, Toru KONO, Yuji OMIYA.
Application Number | 20160008385 14/865845 |
Document ID | / |
Family ID | 40985143 |
Filed Date | 2016-01-14 |
United States Patent
Application |
20160008385 |
Kind Code |
A1 |
KONO; Toru ; et al. |
January 14, 2016 |
ADRENOMEDULLIN PRODUCTION ENHANCER
Abstract
It is an object of the invention to discover a substance that
effectively increases the production of adrenomedullin, as well as
to provide an adrenomedullin production-enhancing agent utilizing
this substance. The adrenomedullin production-enhancing agent is
characterized by inclusion of a ginsenoside, a sanshool, and/or a
shogaol as active ingredients.
Inventors: |
KONO; Toru; (Asahikawa-shi,
JP) ; KANEKO; Atsushi; (Inashiki-gun, JP) ;
OMIYA; Yuji; (Inashiki-gun, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
National University Corporation ASAHIKAWA MEDICAL COLLEGE
TSUMURA & Co. |
Asahikawa-shi
Tokyo |
|
JP
JP |
|
|
Assignee: |
National University Corporation
ASAHIKAWA MEDICAL COLLEGE
Asahikawa-shi
JP
TSUMURA & Co.
Tokyo
JP
|
Family ID: |
40985143 |
Appl. No.: |
14/865845 |
Filed: |
September 25, 2015 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
12918437 |
Aug 19, 2010 |
9193756 |
|
|
PCT/JP2008/052760 |
Feb 19, 2008 |
|
|
|
14865845 |
|
|
|
|
Current U.S.
Class: |
514/26 ;
514/171 |
Current CPC
Class: |
A61P 1/16 20180101; C07J
17/005 20130101; A61K 36/75 20130101; A61P 1/04 20180101; A61P
13/00 20180101; A61K 31/704 20130101; A61P 9/12 20180101; A61P 9/00
20180101; A61K 31/575 20130101; A61K 31/164 20130101; A61K 36/258
20130101; A61K 36/75 20130101; A61P 19/00 20180101; A61P 43/00
20180101; A61K 31/12 20130101; C07J 9/00 20130101; A61P 11/00
20180101; A61P 1/00 20180101; A61P 29/00 20180101; A61K 36/9068
20130101; A61K 36/9068 20130101; A61K 2300/00 20130101; A61P 15/00
20180101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/704 20060101
A61K031/704; A61K 31/164 20060101 A61K031/164; A61K 36/9068
20060101 A61K036/9068; A61K 36/258 20060101 A61K036/258; A61K 36/75
20060101 A61K036/75; A61K 31/575 20060101 A61K031/575; A61K 31/12
20060101 A61K031/12 |
Claims
1. A method of treating hepatitis in a subject in need thereof
comprising, administering to the subject an effective amount of an
agent comprising: a compound represented by formula (1):
##STR00007## wherein R.sup.1, R.sup.2, and R.sup.3, which may be
identical or different, each represent a hydrogen atom, a hydroxyl
group, --O-Glc, --O-Glc-Glc, --O-Glc-Ara, or --O-Glc--Rha, wherein
Glc represents a glucose residue, Ara represents an arabinose
residue, and Rha represents a rhamnose residue; and a compound
represented by formula (2): ##STR00008## wherein R.sup.4 represents
a hydrogen atom or a hydroxyl group, m is 1 or 2, and a wavy bond
line indicates either a Z- or E-configuration; and/or a compound
represented by formula (3): ##STR00009## wherein n is 4, 6, or
8.
2. The method according to claim 1, wherein the compound
represented by formula (1) is at least one ginsenoside selected
from the group consisting of ginsenoside Rb.sub.1, ginsenoside
Rb.sub.2, ginsenoside Rc, ginsenoside Rd, ginseonside Re,
ginsenoside Rg.sub.1, ginsenoside Rg.sub.2, and ginsenoside
Rh.sub.1.
3. The method agent according to claim 1, wherein the compound
represented by formula (2) is at least one sanshool selected from
the group consisting of .alpha.-sanshool, .beta.-sanshool,
.gamma.-sanshool, hydroxy-.alpha.-sanshool,
hydroxy-.beta.-sanshool, and hydroxy-.gamma.-sanshool.
4. The method according to claim 1, wherein the compound
represented by formula (3) is at least one compound selected from
the group consisting of 6-shogaol, 8-shogaol, and 10-shogaol.
5. A method of treating hepatitis in a subject in need thereof
comprising, administering to the subject an effective amount of an
agent comprising: a crude drug comprising the compound represented
by formula (1): ##STR00010## wherein R.sup.1, R.sup.2, and R.sup.3,
which may be identical or different, each represent a hydrogen
atom, a hydroxyl group, --O-Glc, --O-Glc-Glc, --O-Glc-Ara, or
--O-Glc-Rha, wherein Glc represents a glucose residue, Ara
represents an arabinose residue, and Rha represents a rhamnose
residue; and a crude drug comprising the compound represented by
formula (2): ##STR00011## wherein R.sup.4 represents a hydrogen
atom or a hydroxyl group, m is 1 or 2, and a wavy bond line
indicates either a Z- or E-configuration; and/or a crude drug
comprising the compound represented by formula (3): ##STR00012##
wherein n is 4, 6, or 8.
6. The method according to claim 5, wherein the crude drug
comprising the compound represented by formula (1) is ginseng.
7. The method according to claim 5, wherein the crude drug
comprising the compound represented by formula (2) is Japanese
pepper.
8. The method according to claim 5, wherein the crude drug
containing the compound represented by formula (3) is dried ginger
rhizome.
9. A method of treating hepatitis in a subject in need thereof
comprising, administering to the subject an effective amount of an
agent comprising: a Kampo medicine prescription comprising: a crude
drug comprising the compound represented by formula (1);
##STR00013## wherein R.sup.1, R.sup.2, and R.sup.3, which may be
identical or different, each represent a hydrogen atom, a hydroxyl
group, --O-Glc, --O-Glc-Glc, --O-Glc-Ara, or --O-Glc-Rha, wherein
Glc represents a glucose residue, Ara represents an arabinose
residue, and Rha represents a rhamnose residue; and a crude drug
comprising the compound represented by formula (2): ##STR00014##
wherein R.sup.4 represents a hydrogen atom or a hydroxyl group, m
is 1 or 2, and a wavy bond line indicates either a Z- or
E-configuration; and/or a crude drug comprising the compound
represented by formula (3): ##STR00015## wherein n is 4, 6, or
8.
10. The method according to claim 9, wherein the Kampo medicine
prescription is Daikenchutou.
11-20. (canceled)
Description
TECHNICAL FIELD
[0001] The present invention relates to an adrenomedullin
production-enhancing agent, and more particularly to an
adrenomedullin production-enhancing agent that is effective in
preventing and treating diseases such as Crohn's disease by
promoting the production of adrenomedullin, an intestinal peptide
which increases blood flow and has anti-inflammatory effects.
BACKGROUND ART
[0002] Adrenomedullin was discovered in 1993 as a peptide involved
in regulation of the circulatory system with strong vasodilatory
effects (Patent Document 1). Adrenomedullin is produced by a
variety of organs such as those of the circulatory system and the
digestive system, and has important physiological effects such as
vasodilatation, neovascularization, antibacterial effects,
anti-enteritic effects, protection of the gastric mucosa, and
suppression of thrombus formation. Administration of adrenomedullin
has correspondingly been confirmed to be effective in the treatment
of various diseases, and has been reported to be effective against
conditions including myocardial disorders, non-bacterial
inflammatory diseases, pulmonary hypertension, bone disorders,
myometrial contraction, urinary disorders, and the like (Patent
Documents 2 to 7).
[0003] However, since adrenomedullin is a peptide, formulation of
it is associated with high costs of production, and in the case of
direct administration of adrenomedullin, a dosage form such as an
injectable preparation or an intravenous drip preparation must be
used. Rigorous production technology and management excluding
incorporation of foreign matter, such as endotoxin, is thus
required. Furthermore, since adrenomedullin has a half-life in
blood of only tens of minutes, administration of significant
amounts is required for exertion of its effects. However,
intravenous administration of a large amount of adrenomedullin may
induce hypotension. Moreover, when exposure of specific organs to
adrenomedullin is expected, various problems may exist, such as the
need for targeting technologies. There has thus been a need for a
highly safe medicament which can be produced at low cost and that
is when orally administered capable of enhancing activity of the
adrenomedullin production system, which is constitutively activated
in vivo. However, no such medicament has been available.
[0004] Patent Document 1: Japanese Patent No. 2774769
[0005] Patent Document 2: WO 00/078338
[0006] Patent Document 3: WO 00/078339
[0007] Patent Document 4: JP-T-2002-540216
[0008] Patent Document 5: JP-A-2003-300899
[0009] Patent Document 6: JP-A-2006-290777
[0010] Patent Document 7: JP-A-2006-290814
DISCLOSURE OF THE INVENTION
Problems to be Solved by the Invention
[0011] It is thus an object of the present invention to determine a
substance that can be orally administered and that effectively
enhances the production of adrenomedullin in vivo, and in addition
to provide an adrenomedullin production-enhancing agent utilizing
this substance.
Means for Solving the Problems
[0012] In an attempt to solve the above-described problems, the
inventors of the present invention have searched extensively for
substances which enhance adrenomedullin production, and have found
that an excellent adrenomedullin production-enhancing effect is
obtained by combining specific compounds that are contained in
known crude drugs, thus eventually completing the invention.
[0013] Specifically, the present invention is an adrenomedullin
production-enhancing agent containing, as active ingredients, a
compound represented by the following formula (1):
##STR00001##
wherein R.sup.1, R.sup.2, and R.sup.3, which may be identical or
different, each represent a hydrogen atom, a hydroxyl group,
--O-Glc, --O-Glc-Glc, --O-Glc-Ara, or --O-Glc-Rha, provided that
Glc represents a glucose residue, Ara an arabinose residue, and Rha
a rhamnose residue; and
[0014] a compound represented by the following formula (2) and/or a
compound represented by formula (3):
##STR00002##
wherein R.sup.4 represents a hydrogen atom or a hydroxyl group; m
is 1 or 2; and the wavy line indicates whether the part is in the
Z- or E-configuration;
##STR00003##
wherein n is 4, 6, or 8.
Effects of the Invention
[0015] The adrenomedullin production-enhancing agent of the
invention effectively promotes the production of adrenomedullin,
which has various physiological effects including increasing blood
flow as well as anti-inflammatory effects. The adrenomedullin
production-enhancing agent of the invention can thus be used in the
treatment of various diseases. For example, by increasing the
amount of blood flow inside the intestinal tract, Crohn's disease,
an ischemic disease of the digestive system, and the like can be
effectively prevented or treated by it. Furthermore, since the
adrenomedullin production-enhancing agent originates from natural
substances and can be orally administered, it is very safe,
exhibits excellent sustainability of effects, and strongly enhances
adrenomedullin production, particularly in the digestive tract.
Best Mode for Carrying Out the Invention
[0016] As an active ingredient of the adrenomedullin
production-enhancing agent of the invention, a compound represented
by the following formula (1) (hereinafter, may be referred to as
"compound (1)") is used:
##STR00004##
wherein R.sup.1, R.sup.2, and R.sup.3, which may be identical or
different, each represent a hydrogen atom, a hydroxyl group,
--O-Glc, --O-Glc-Glc, --O-Glc-Ara, or --O-Glc-Rha, provided that
Glc represents a glucose residue, Ara an arabinose residue, and Rha
a rhamnose residue.
[0017] Concerning compound (1), specifically, there may be
mentioned ginsenosides such as ginsenoside Rb.sub.1, in which
R.sup.1 and R.sup.2 are each --O-Glc-Glc and R.sup.3 is a hydrogen
atom; ginsenoside Rb.sub.2 or Rc, in which R.sup.1 is --O-Glc-Glc,
R.sup.2 is --O-Glc-Ara, and R.sup.3 is a hydrogen atom; ginsenoside
Rd, in which R.sup.1 is --O-Glc-Glc, R.sup.2 is --O-Glc, and
R.sup.3 is a hydrogen atom; ginsenoside Re, in which R.sup.1 is a
hydroxyl group, R.sup.2 is --O-Glc, and R.sup.3 is --O-Glc-Rha;
ginsenoside Rg.sub.1, in which R.sup.1 is a hydroxyl group, R.sup.2
is --O-Glc, and R.sup.3 is --O-Glc; ginsenoside Rg.sub.2, in which
R.sup.1 and R.sup.2 are each a hydroxyl group and R.sup.3 is
--O-Glc-Rha; and ginsenoside Rh.sub.1, in which R.sup.1 and R.sup.2
are each a hydroxyl group and R.sup.3 is --O-Glc. Among these,
ginsenoside and ginsenoside Rg.sub.1 are preferred.
[0018] These compounds can be isolated by known methods (document:
Kampo Medicine, Vol. 35, No. 1, Pages 1-22 (1984) , and the like),
or commercially available products can be used.
[0019] Furthermore, the adrenomedullin production-enhancing agent
of the invention uses the above-described compound represented by
formula (1) , in combination with a compound represented by the
following formula (2) (hereinafter, may be referred to as "compound
(2)") and/or a compound represented by formula (3) (hereinafter,
may be referred to as "compound (3)").
##STR00005##
wherein R.sup.4 represents a hydrogen atom or a hydroxyl group, m
is 1 or 2, and the wavy line indicates whether the part is in the
Z- or E-configuration;
##STR00006##
wherein n is 4, 6, or 8.
[0020] Concerning compound (2), specifically, there may be
mentioned sanshools such as .alpha.-sanshool, in which R.sup.4 is a
hydrogen atom, m=1, and the wavy line is in the Z-configuration;
.beta.-sanshool, in which R.sup.4 is a hydrogen atom, m=1, and the
wavy line is in the E-configuration; y-sanshool, in which R.sup.4
is a hydrogen atom, m=2, and the wavy line is in the
Z-configuration; hydroxy-.alpha.-sanshool, in which R.sup.4 is a
hydroxyl group, m=1, and the wavy line is in the Z-configuration;
hydroxy-.beta.-sanshool, in which R.sup.4 is a hydroxyl group, m=1,
and the wavy line is in the E-configuration; and
hydroxy-.gamma.-sanshool, in which R.sup.4 is a hydroxyl group,
m=2, and the wavy line is in the Z-configuration. Among these,
hydroxy-.alpha.-sanshool and hydroxy-.beta.-sanshool are
preferred.
[0021] These compounds can be isolated or synthesized by known
methods (document: Biosci Biotechnol Biochem: Vol. 69, No. 10,
1951-1957 (2005); Biological & Pharmaceutical Bulletin (2007),
30(1), 205-207; Phytochemistry (1997), 44(6), 1125-1127; and the
like).
[0022] On the other hand, concerning compound (3), specifically,
6-shogaol, 8-shogaol, 10-shogaol, and the like may be mentioned.
Among these, 6-shogaol is preferred.
[0023] These compounds can be isolated or synthesized by known
methods (document: Bulletin of the Chemical Society of Japan
(1976), 49(5), 1453-1454; Japanese Patent Application No.
63-137843; and the like), or commercially available products can be
used.
[0024] The adrenomedullin production-enhancing agent of the
invention can be produced using the compound represented by formula
(1) and the compound represented by formula (2) and/or the compound
represented by formula (3) as active ingredients, appropriately
mixing these compounds with other pharmaceutical carriers, and
formulating this mixture into an oral or a parenteral
preparation.
[0025] Oral preparations in the form of dust, powders, granules,
tablets, capsules, soft capsules, liquids, and the like can be
formulated, and pharmaceutical carriers adequate for these, for
example starch, lactose, sucrose, mannite, carboxymethylcellulose,
corn starch, inorganic salts, and the like, can be used. Upon
production of an oral preparation, a binder, a disintegrant, a
surfactant, a lubricating agent, a fluidity-promoting agent, a
flavoring agent, a colorant, a fragrance, and the like may be
incorporated.
[0026] Furthermore, parenteral preparations can also be produced
according to conventional methods, and distilled water for
injection, physiological saline, aqueous glucose solution, plant
oil for injection, sesame oil, peanut oil, soybean oil, corn oil,
propylene glycol, polyethylene glycol, and the like can generally
be used as diluents. If necessary, a bactericidal agent, a
preservative, a stabilizer, and the like can be further added.
[0027] In regard to the adrenomedullin production-enhancing agent
of the invention, the amount of compound (1) to be incorporated as
an active ingredient varies with the type of compound, disease to
be treated, severity of the disease, age of the patient, and the
like, though, for example, in the case of use of ginsenoside
Rb.sub.1, the amount to be incorporated is about 1 mg to 1 g as a
daily dose for an adult. In the case of use of compound (2) as the
component in combination with compound (1), the amount to be
incorporated is about 1 mg to 1 g as a daily dose for an adult. In
the case of use of compound (3) as the component in combination
with compound (1), the amount to be incorporated is about 1 mg to 1
g as a daily dose for an adult. On the other hand, in the case of
use of compound (2) and compound (3) in combination, the amount to
be incorporated is about 2 mg to 2 g in total.
[0028] In regard to the adrenomedullin production-enhancing agent
of the invention, it is also possible to use a crude drug
containing compound (1), and a crude drug containing compound (2)
and/or a crude drug containing compound (3) as active ingredients.
As a crude-drug containing compound (1), ginseng may be mentioned.
As a crude drug containing compound (2), Japanese pepper (Sansho)
may be mentioned, and as a crude drug containing compound (3),
dried ginger rhizome (Kankyo) may be mentioned. Ginseng, Japanese
pepper, and dried ginger rhizome are all materials known as Kampo
medicine ingredients, and commercially available products can also
be used.
[0029] Preparation of the adrenomedullin production-enhancing agent
of the invention is carried out by chopping or pulverizing the
aforementioned ginseng, and Japanese pepper and/or dried ginger
rhizome, separately, and then mixing them uniformly, or if
necessary by drying an extract obtained from the materials with an
appropriate solvent, and then mixing the resulting product with
Saccharum granorum as necessary.
[0030] The amount of ginseng to be incorporated in the
adrenomedullin production-enhancing agent of the invention is about
1 mg to 1 g as a daily dose for an adult. In the case of use of
Japanese pepper as the component in combination with ginseng, the
amount to be incorporated is about 1 mg to 1 g, and in the case of
use of dried ginger rhizome, the amount to be incorporated is about
1 mg to 1 g. On the other hand, in the case of use of Japanese
pepper and dried ginger rhizome in combination, the amount to be
incorporated is about 2 mg to 2 g in total.
[0031] Examples of Kampo medicine prescriptions containing ginseng
and Japanese pepper and/or dried ginger rhizome include
Daikenchutou, Toukitou, Hangeshashintou, Ninjintou,
Hangebyakujyutsutenmatou, Hochuekkitou, Keishinninjintou,
Daiboufutou, Ourentou, and the like, and these can also be used as
active ingredients for the adrenomedullin production-enhancing
agent of the invention. Taking the general constitution of
Daikenchutou as an example, 15 g of Daikenchutou contains 1.25 g of
a dried extract of mixed crude drugs in proportions of dried ginger
rhizome:ginseng:Japanese pepper=5:3:2 (powdered Daikenchutou
extract), and 10 g of Saccharum granorum.
[0032] The adrenomedullin production-enhancing agent of the
invention can be produced using the aforementioned ginseng and
dried ginger rhizome and/or Japanese pepper as active ingredients,
mixing them with other pharmaceutical carriers, and formulating the
mixture obtained into an oral or a parenteral preparation. The same
components as described above can be used as pharmaceutical
carriers.
[0033] When the adrenomedullin production-enhancing agent of the
invention that is obtainable as described above is administered,
adrenomedullin production by various organs and tissues is
promoted. Since adrenomedullin has various physiological effects,
such as vasodilatation, neovascularization, antibacterial activity,
anti-enteritic effects, protection of the gastric mucosa, and
suppression of thrombus formation, it is effective in the
prevention and treatment of diseases including inflammatory bowel
diseases such as ulcerative colitis and Crohn' s disease, urinary
disorders, myometrial contraction, bone disorders, hypertension,
myocardial disorders, non-bacterial inflammatory diseases, and
hepatitis.
EXAMPLES
[0034] Hereinafter, the present invention will be explained in
greater detail by way of Examples, though the invention is not at
all intended to be limited to these Examples. In the Examples, a
powdered extract obtained by extracting crude drug or a mixture of
crude drugs with water according to a conventional method was
used.
Example 1
[0035] Adrenomedullin Production-Enhancing Effect Of
Daikenchutou:
[0036] Male SD rats (8 to 10 weeks of age, body weight 300 to 400
g) were used (n=16). Saccharum granorum was added to distilled
water to a concentration of 480 mg/mL to prepare an aqueous
solution of it. To this aqueous solution of Saccharum granorum, a
powdered extract of Daikenchutou was added to a concentration of 60
mg/mL, by weighing immediately at the time of use, and the mixture
was homogeneously dispersed by stirring for 30 minutes at room
temperature for use as a test specimen. A 5 mL/kg portion of the
test specimen, which had been kept warm at 37.degree. C., was
administered into the duodenum through a cannula, and blood was
collected from the portal vein at 0, 15, 30, 60, and 90 minutes
after administration. A control group was administered distilled
water in the same fashion. Approximately 5 mL of the portal vein
blood was inserted into a 15-mL centrifuge tube made of
polypropylene (PP), in which an ethylenediaminetetraacetic acid
(EDTA)/aprotinin solution (containing 5 mg of EDTA-2Na and 2500 KIU
of aprotinin) had been dispensed in an amount of 100 .mu.L, and
centrifugation was performed at 1500 g at 4.degree. C. for 15
minutes. Two mL of blood plasma and 0.16 mL of an acidified
solution having the following composition were added to a 5-mL
centrifuge tube made of PP for high-speed centrifuge to acidify the
plasma sample.
[0037] The supernatant obtained by centrifugation at 7000 g at
4.degree. C. for 20 minutes was passed through an activated Sep-Pak
cartridge column (C-18 column, WAT020805 manufactured by Waters
Corp.), and adrenomedullin (ADM) was adsorbed to the column. The
column was washed twice with 2.5 mL of a column washing liquid
(0.1% aqueous solution of trifluoroacetic acid (TFA)), and the
adsorbed ADM was then eluted with 2 mL of a column eluent (0.1% TFA
methanol solution). The eluate was received into a 5-mL centrifuge
tube and subjected to centrifugation under reduced pressure to
evaporate and dry it to a solid. The dried sample was stored at
-80.degree. C. until EIA (Enzyme immunoassay) measurement.
Quantification of ADM was carried out using a Rat ADM EIA kit
(EK-010-08, manufactured by Phoenix Pharmaceuticals, Inc.). The
evaporated and dried solid sample was dissolved in 400 .mu.L, of an
EIA buffer solution, and the solution was heated at 90.degree. C.
for 15 minutes and then centrifuged at 2000 g for 20 minutes. The
resulting supernatant was used as the EIA sample. Data are
expressed as the mean value.+-.standard error (S.E.M.), and were
subjected to two-way ANOVA and then to Dunnett's multiple
comparison test or Student's t-test. A risk rate of 5% or less was
employed as the significance level. Changes in ADM concentration in
blood plasma are presented in Table 1 and FIG. 1.
[0038] (Composition of acidified solution)
TABLE-US-00001 NaCl 100 mg (final concentration: 1%)
Trifluoroacetic acid (TFA) 0.1 mL (final concentration: 1%) Formic
acid 0.5 mL (final concentration: 5%) 1N hydrochloric acid 8.0 mL
(final concentration: 80%) Distilled water 1.4 mL Total: 10 mL
TABLE-US-00002 TABLE 1 ADM concentration (pg/mL) 0 min 15 min 30
min 60 min 90 min Control Mean 30.4 33.2 37.4 38.7 38.6 value
Standard 2.7 3.0 3.3 3.0 4.6 error Daikenchutou Mean 30.4 43.3 54.3
53.0 44.1 value Standard 2.7 3.0 3.7 4.4 3.3 error
[0039] While the ADM concentration in plasma of portal vein blood
recovered immediately before enteric injection of the test specimen
was 30.4.+-.2.7 pg/mL, the ADM concentrations for recovery at 15,
30, 60, and 90 minutes after administration of the test specimen
Daikenchutou were 43.3.+-.3.0, 54.3.+-.3.7, 53.0.+-.4.4, and
44.1.+-.3.3 pg/mL, respectively, and significantly increased over
time. In a comparison between the group administered distilled
water and that administered Daikenchutou, significant differences
in ADM concentration (p<0.01) were confirmed at 15, 30, and 60
minutes after administration.
Example 2
[0040] Blood Flow-Increasing Effects Of Daikenchutou And Its
Constituent Crude Drugs:
[0041] Male SD rats (9 to 11 weeks of age, body weight 260 to 350
g) were used (n=3 or 6). Saccharum granorum and a powdered extract
of Daikenchutou were suspended in distilled water to obtain a
concentration of 160 mg/mL of Saccharum granorum and to a
concentration of 20 mg/mL for the powdered extract of Daikenchutou,
and the suspension was used as a test specimen (Daikenchutou).
Furthermore, test specimens respectively containing the individual
crude drugs were prepared by adding the ginseng extract powder,
dried ginger rhizome extract powder, and Japanese pepper extract
powder to distilled water to obtain concentrations of 6 mg/mL, 10
mg/mL, and 4 mg/mL, respectively. Furthermore, test specimens
containing two crude drugs among the aforementioned three crude
drugs in combination were prepared by addition to distilled water
to obtain concentrations of 6 mg/mL for the ginseng extract powder
and 10 mg/mL for the dried ginger rhizome extract powder, 6 mg/mL
for the ginseng extract powder and 4 mg/mL for the Japanese pepper
extract powder, or 4 mg/mL of the Japanese pepper extract powder
and 10 mg/mL of the dried ginger rhizome extract powder. After
preliminarily placing an 18G Surf lo indwelling needle inserted
through the caecum, 5 mL/kg portions of test specimens which had
been kept warm at 37.degree. C. were administered into the colon
0.5 to 1 hour later, by which time baseline blood flow had
stabilized.
[0042] A polyethylene tube with an inner diameter of 0.58 mm was
inserted through the left common carotid artery of an anesthetized
rat, and the tip was left indwelling in the left ventricle. A
tracheal cannula was placed indwelling in the trachea and then
connected to a respirator (SN-480-7, manufactured by Shinano
Seisakusho Co., Ltd.); artificial respiration was then performed at
a frequency of 60 RPM. Body temperature was maintained at
37.+-.0.5.degree. C. by a temperature controller (NS-TC10,
manufactured by Neuroscience, Inc.). The rat was incised along the
midline of the abdomen, the caecum was taken outside the body, and
then the distal colon was exteriorized. The lower part of the colon
was lightly lifted with forceps, and with the feces still present
inside the colon, four sites were tied and fixed to the end of the
incised right and left rectus abdominis with 5-0 suture, such that
the length between the tied sites was 1 cm. A blood flow probe was
positioned at the upper part of the fixed distal colon, and blood
flow (flow, mass, velocity) was measured using a laser tissue blood
flow meter (ALF21N, manufactured by Advance Co., Ltd.). To prevent
drying, the entire abdomen including the blood flow probe was
covered with plastic wrap. In addition, monitoring of blood
pressure and heart rate was carried out simultaneously with amount
of blood flow, and the respective results of measurement were
recorded with data analysis software (Chart V3.6 [v5.4.2],
manufactured by ADInstruments Corp.) via a patient monitoring
apparatus (BP-508, manufactured by Colin Corp.) [biological
amplifier (Nihon Kohden Corp.)] and a data recording apparatus
(Power Lab/800 [8/30], manufactured by ADInstruments Corp.). The
blood flow measurement was continued until 90 minutes after drug
administration, vascular conductance (VC) obtained by dividing flow
by blood pressure was employed as an index for blood flow, and the
rate of increase (%) in VC was calculated. The rates of increase in
VC for the respective test specimens are presented in Table 2 and
FIGS. 2 to 8.
TABLE-US-00003 TABLE 2 Rates of increase in VC for respective test
specimens Test specimen VC (%) (mg/kg) 15 min 30 min 45 min 60 min
75 min 90 min Daikenchutou Mean 30.23 26.51 42.70 73.33 99.00
100.86 (900) value Standard 13.83 9.00 8.42 7.77 15.93 36.45 error
Ginseng (30) Mean 69.37 21.85 2.03 20.50 20.23 17.40 value Standard
25.67 3.49 14.61 19.39 24.58 32.38 error Dried ginger Mean 8.84
13.42 19.51 11.51 18.22 33.42 rhizome (50) value Standard 6.56 6.96
9.72 8.18 7.06 20.07 error Japanese Mean 13.55 25.84 33.95 50.65
76.63 40.80 pepper (20) value Standard 6.72 19.94 11.81 23.06 23.71
29.70 error Ginseng (30) Mean 46.57 20.67 31.65 67.86 64.13 69.72
Dried ginger value rhizome (50) Standard 6.95 5.93 6.94 20.12 15.48
13.80 error Ginseng (30) Mean 55.89 40.74 69.16 111.18 139.12
124.43 Japanese value pepper (20) Standard 7.91 17.89 28.20 27.69
30.45 9.36 error Dried ginger Mean 9.60 21.56 35.76 72.89 87.30
87.48 rhizome (50) value Japanese Standard 8.85 9.68 10.19 17.21
24.34 24.21 pepper (20) error
[0043] The group administered Daikenchutou exhibited an increase in
VC at 15 to 90 minutes after administration. Among the groups
administered individual constituent crude drugs of Daikenchutou,
that administered ginseng exhibited a temporary increase in blood
flow at 15 minutes after administration, and the group administered
Japanese pepper exhibited a weak increase, with a peak at 75
minutes after administration. Dried ginger rhizome did not clearly
increase blood flow. Among the combinations of constituent crude
drugs, that of ginseng with dried ginger rhizome and that of
ginseng with Japanese pepper yielded clearly higher increases in VC
than administrations of the individual components alone. No change
in the rate of increase was observed with combined use of dried
ginger rhizome and Japanese pepper.
Example 3
[0044] Effect Of ADM Antagonist Pretreatment On The Blood
Flow-Increasing Effect Of Daikenchutou:
[0045] Male SD rats were divided into two groups: a
Daikenchutou-treated group and an ADM antagonist-pretreated group
(n=7 or 8). A solution with a 160 mg/mL concentration of Saccharum
granorum was prepared using distilled water, and powdered
Daikenchutou extract was weighed at the time of use to obtain a
concentration of 20 mg/mL and homogeneously dispersed in the same
solution by stirring at room temperature for 30 minutes or longer
to obtain Daikenchutou (TJ-100). This was administered into the
colon in the Daikenchutou-treated group in an amount of 900 mg/5
mL/kg. On the other hand, for the ADM antagonist-pretreated group,
a human ADM antagonist (h. ADM22-52, 4302-v, manufactured by
Peptide Institute, Inc.) was dissolved in physiological saline at
30 .mu.mol/L, and the solution was intravenously administered
through a cannula placed under anesthesia, at a dose of 30 nmol/l
mL/kg, and 15 minutes later Daikenchutou was administered into the
colon in an amount of 900 mg/5 mL/kg.
[0046] A polyethylene tube with an inner diameter of 0.8 mm was
inserted through the left common carotid artery of an anesthetized
rat, and a tip was left indwelling in the left ventricle. A
tracheal cannula was placed indwelling in the trachea and connected
to a respirator (SN-480-7); artificial respiration was then
performed at a frequency of 60 RPM. Body temperature was maintained
at 37.+-.0.5.degree. C. by a temperature controller (NS-TC10). The
rate of increase in VC (%) was calculated in the same fashion as in
Example 2. The rates of increase in VC for the respective groups
are presented in Table 3 and FIG. 9. The results of measurement are
expressed as the mean value.+-.standard error (S.E.M.) . Comparison
of effects was performed by one-way analysis of variance (ANOVA),
followed by the Scheffe multiple comparison test. A risk rate of 5%
or less was employed as the significance level.
TABLE-US-00004 TABLE 3 Suppression by ADM antagonist of increase in
blood flow by Daikenchutou VC (%) Test specimen 15 min 30 min 45
min 60 min 75 min 90 min Daikenchutou Mean 39.82 34.79 70.15 95.29
106.80 96.81 (900 mg/kg) value Standard 7.66 7.18 13.16 14.45 14.94
19.40 error Daikenchutou Mean 8.72 4.03 4.07 1.28 7.28 15.12 (900
mg/kg) + value ADM(22-52) (30 Standard 10.85 9.40 8.71 9.75 8.53
12.95 nmol/kg) error
[0047] With ADM antagonist pretreatment, a significant decrease in
VC of 30.8 to 99.5% compared to the group administered Daikenchutou
alone was found up to 30 to 90 minutes after administration. Since
ADM antagonist pretreatment decreased the increase in blood flow
induced by Daikenchutou, it was confirmed that ADM has a blood
flow-increasing effect.
Example 4
[0048] Blood Flow-Increasing Effects Of Components In Crude Drugs
(1):
[0049] Male SD rats (8 to 10 weeks of age, body weight 240 to 360
g) were used (n=2 to 6) . Hydroxy-.beta.-sanshool, ginsenoside
Rb.sub.1, or ginsenoside Rg.sub.1 was added to a 1% aqueous
solution of Tween 80 to obtain concentrations of 0.06 mg/mL, 0.2
mg/mL, and 0.2 mg/mL, respectively, and the resulting solutions
were used as the respective test specimens for administration of
individual crude drug components. Furthermore, test specimens for
the administration of hydroxysanshool with a ginseng extract,
ginsenoside Rb.sub.1, or ginsenoside Rg.sub.1 in combination were
prepared by adding, to a 1% aqueous solution of Tween 80,
hydroxysanshool at a concentration of 0.06 mg/mL, and a powdered
ginseng extract, ginsenoside Rb.sub.1, or ginsenoside Rg.sub.1 at a
concentration of 6 mg/mL, 0.2 mg/mL, or 0.2 mg/mL, respectively.
After preliminarily placing an 18G Surflo indwelling needle
inserted through the caecum, 5 mL/kg portions of test specimens
which had been kept warm at 37.degree. C. were administered into
the colon 0.5 to 1 hour later, by which time baseline blood flow
had stabilized. A control group was administered a 1% aqueous
solution of Tween 80.
[0050] A polyethylene tube with an inner diameter of 0.8 mm was
inserted through the left common carotid artery of an anesthetized
rat, and the tip was left indwelling in the left ventricle. A
tracheal cannula was placed indwelling in the trachea and connected
to a respirator (SN-480-7); artificial respiration was then
performed at a frequency of 60 RPM. Body temperature was maintained
at 37.+-.0.5.degree. C. by a temperature controller (NS-TC10). The
rate of increase in VC (%) was calculated in the same fashion as in
Example 2. The rates of increase in VC for the respective test
specimens are presented in Table 4 and FIGS. 10 to 12. The results
of measurement are expressed as the mean value .+-.standard error
(S.E.M.). Comparison of effects was performed by one-way analysis
of variance (ANOVA), followed by the Scheffe multiple comparison
test. A risk rate of 5% or less was employed as the significance
level.
TABLE-US-00005 TABLE 4 Rates of increase in VC by hydroxysanshool
Test specimen VC (%) (mg/kg) 15 min 30 min 45 min 60 min 75 min 90
min Control Mean -17.77 -22.20 -25.23 -28.48 -18.04 -16.61 value
Standard 9.58 7.12 5.71 6.71 8.01 4.36 error H-sanshool Mean 3.89
5.79 2.09 9.46 8.58 4.19 (0.3) value Standard 6.08 6.68 8.04 7.88
8.42 8.10 error H-sanshool Mean 3.41 31.49 48.67 73.02 90.97 57.58
(0.3) value Ginseng (30) Standard 7.35 15.99 12.10 17.43 38.59
21.18 error Ginsenoside Mean 11.72 18.87 60.21 26.20 40.81 19.98
Rb.sub.1 (1) value Standard 4.36 17.59 7.74 0.00 2.68 0.39 error
H-sanshool Mean -13.97 22.26 50.64 87.81 114.10 104.80 (0.3) +
value Ginsenoside Standard 17.67 39.88 37.38 34.76 29.22 50.72
Rb.sub.1 (1) error Ginsenoside Mean -10.83 5.84 16.29 10.92 12.25
18.11 Rg.sub.1 (1) value Standard 14.14 25.96 5.66 10.58 1.85 4.96
error H-sanshool Mean 76.79 71.25 84.85 122.22 139.55 165.77 (0.3)
+ value Ginsenoside Standard 61.48 72.71 85.25 77.74 82.35 87.90
Rg.sub.1 (1) error
[0051] Hydroxysanshool did not increase blood flow when
administered alone. However, when used in combination with the
ginseng extract, a significant increase in VC of 53.7 to 109.0% was
observed compared to the control group up to 30 to 90 minutes after
administration. Furthermore, compared with the group administered
hydroxysanshool alone, the combination yielded significant
increases in VC of 45.4, 63.4, and 46.8%, respectively, at 45, 60,
and 90 minutes after administration. In the case of combined use,
peak increase appeared at 75 minutes after administration,
consistent with the pattern of blood flow increase observed in the
case of administration of Daikenchutou in Example and single
administration of a Japanese pepper extract. Furthermore, the group
administered hydroxysanshool and ginsenoside Rb.sub.1 or Rg.sub.1
in combination also exhibited marked increases in VC compared to
the group administered hydroxysanshool alone. These findings
indicated that hydroxysanshool functions as an active ingredient of
Japanese pepper, and suggested that ginseng plays an important role
in expression of the activity of hydroxysanshool.
Example 5
[0052] Blood Flow-Increasing Effects Of Components In Crude Drugs
(2):
[0053] Male SD rats (8 to 10 weeks of age) were used. 6-Shogaol was
added to a 1% aqueous solution of Tween 80 to obtain a
concentration of 0.4 mg/mL, and the resulting solution was used as
a test specimen. Furthermore, ginsenoside Rb.sub.1 and 6-shogaol
were added to a 1% aqueous solution of Tween 80 at concentrations
of 0.2 mg/mL and 0.4 mg/mL, respectively, to prepare test
specimens. After preliminarily placing an 18G Surf lo indwelling
needle inserted through the caecum, 5 mL/kg portions of test
specimens which had been kept warm at 37.degree. C. were
administered into the colon 0.5 to 1 hour later, by which time
baseline blood flow had stabilized. A control group was
administered 1% aqueous solution of Tween 80.
[0054] A polyethylene tube with an inner diameter of 0.8 mm was
inserted through the left common carotid artery of an anesthetized
rat, and the tip was left indwelling in the left ventricle. A
tracheal cannula was placed indwelling in the trachea and connected
to a respirator (SN-480-7); artificial respiration was then
performed at a frequency of 60 RPM. Body temperature was maintained
at 37 .+-.0.5.degree. C. by a temperature controller (NS-TC10). The
rate of increase in VC (%) was calculated in the same fashion as in
Example 2. The rates of increase in VC for the respective test
specimens are presented in Table 5 and FIG. 13.
TABLE-US-00006 TABLE 5 Test specimen VC (%) (mg/kg) 15 min 30 min
45 min 60 min 75 min 90 min 6-Shogaol (2) 58.32 61.20 53.07 42.55
46.68 32.92 6-Shogaol (2) + -14.17 13.84 72.66 43.75 63.84 58.82
Ginsenoside Rb.sub.1 (1)
[0055] 6-Shogaol, when administered alone, induced an increase in
blood flow from just after administration that peaked at 30
minutes. When used in combination with ginsenoside Rb.sub.1, it
yielded an increase in blood flow which peaked at 45 minutes.
Example 6
[0056] Anti-Enteritic Effect In An Animal Model Of Crohn'S
Disease:
[0057] Male BALB/c mice (20 to 25 g) were used (n=6 or 7).
Saccharum granorum and powdered Daikenchutou extract were added to
distilled water to obtain concentrations of 80 mg/mL and 10 mg/mL,
respectively, for preparation of the test specimen
(Daikenchutou).
[0058] 2,4,6-Trinitro-benzene sulfonic acid (TNBS, manufactured by
Tokyo Chemical Industry Co., Ltd.) was weighed in a tube made of
PP, a 50% aqueous solution of EtOH was added to the tube, and the
mixture was stirred and dissolved to obtain a concentration of 15
mg/mL. A 1-mL Terumo syringe was connected to a feeding tube
(SF-FT0380FG, manufactured by Terumo Corp., Fr3.5, outer diameter
1.2 mm). A position 3.5 cm from a tip of the cannula was marked
with an oil marker pen. The mouse was left alone in a small wire
mesh cage for 15 to 30 minutes to induce defecation, and anesthesia
was performed by peritoneal administration of pentobarbital at 55
mg/kg and atropine at 0.75 mg/kg (manufactured by Sigma-Aldrich
Company). An additional 15 to 30 minutes later, a cannula in which
TNBS solution had been filled, was slowly inserted to a depth of
3.5 cm after attaching olive oil at the tip (with olive oil added
to the tip). With the cannula still inserted, the anus was closed
with a paper clip, and the intestinal lumen was sealed. The mouse
was suspended in the air, TNBS was slowly infused (2 sec/0.1 mL) in
an amount of 1.5 mg/0.1 mL/head, and the mouse was maintained as
such for 30 seconds to induce enteritis.
[0059] At 8, 24, 32, 48, and 56 hours after the infusion of TNBS,
Daikenchutou was forcibly administered orally in an amount of 900
mL/kg. A control group was administered distilled water in the same
fashion. The mice were subjected to laparotomy on the third day (72
hours) from the time of induction of enteritis by infusion of TNBS
into the intestine, and the presence or absence of adhesions was
observed. The portion from the caecum to the anus was collected,
and photographs were taken (Digital Camera D100, manufactured by
Nikon Corp.). This portion was cut along the longitudinal muscles,
its content was washed away with physiological saline, and
photographs were taken again, with the mucosal surface facing
upward. The photographic images were inputted into image analysis
software (Image J), and the area of necrosis (cm.sup.2) of the
lumen surface was measured. An untreated group which had not been
subjected to induction of enteritis and administration of the test
specimen was evaluated for disease state in the same fashion.
[0060] Measurements of the area of necrosis were examined for
differences between groups by Welch's t-test. A risk rate of 5% or
less was employed as the significance level. The results of
measurement of area of necrosis are presented in FIG. 14, and the
frequency of adhesions is presented in FIG. 15. Furthermore, a
photograph of the large intestine at 72 hours after infusion of
TNBS into the intestine is presented in FIG. 16.
[0061] In the control group, visual evaluation of the large
intestine three days after induction of enteritis was performed,
and formation of mesenteric adhesions in the large intestine and
severe necrosis of the luminal mucosa were observed. On the other
hand, in the group administered Daikenchutou five times orally from
8 hours after TNBS treatment, the frequency of adhesions in the
large intestine and the area of severe necrosis in the inner lumen
were clearly decreased.
Example 7
[0062] Effects Of Daikenchutou In A Rat Model Of Hepatic
Cirrhosis:
[0063] A thioacetamide (TA) solution at a concentration of 300 mg/L
was administered by feeding to a male SD rat for 20 weeks to induce
hepatic cirrhosis. Powdered Daikenchutou extract was administered
by mixing with feed at a dose of 50 or 200 mg/kg/day, from the 10th
week after initiation of TA treatment to the end of the test. For
evaluation of disease state, blood and hepatic tissues were
collected on the 10th and 20th weeks after the TA treatment. The
collected hepatic tissues were subjected to hematoxylin-eosin (HE)
and Sirius red staining, and the pathologic images were inputted to
image analysis software (Image J) to evaluate hepatic fibrosis in
terms of area ratio. The amount of hyaluronic acid in blood was
measured using a hyaluronic acid ELISA kit (manufactured by
CosmoBio Co., Ltd.). Furthermore, quantification of the amount of
hydroxyproline in liver was performed by the Norman-Logan
method.
[0064] All test results are expressed as the mean value.+-.standard
deviation (S.D.). Statistical analysis was performed using
Dunnett's multiple comparison tests between the groups administered
TA solution for 20 weeks, with a risk rate of 5% or less used as
the significance level.
[0065] The results of evaluation of hepatic fibrosis are presented
in FIG. 17, the results of measurement of the amount of hyaluronic
acid in blood in FIG. 18, and the results of quantification of
hydroxyproline in liver in FIG. 19.
[0066] In the hepatitis control group on the 10th and 20th week
after TA treatment, levels of hyaluronic acid in blood and
hydroxyproline in liver, which are markers of hepatic fibrosis,
were each significantly increased compared to the untreated group,
and increases in them were particularly obvious at the 20th week.
In the hepatitis control group, at the 20th week after TA
treatment, Sirius Red-positive pathological changes were clearly
observed, and findings characteristic of hepatic cirrhosis were
exhibited. On the other hand, in the group administered
Daikenchutou from the 10th week after TA treatment and evaluated at
the 20th week, increases in the levels of hyaluronic acid in blood
and hydroxyproline in liver were significantly less than those in
the control group, with clear manifestation of efficacy. These
effects were confirmed by HE and Sirius Red staining. Daikenchutou
thus inhibited progression of hepatic fibrosis and was clearly
efficacious in a TA-induced model of hepatic cirrhosis.
[0067] Preparation Example 1
[0068] A 50 g portion of ginsenoside Rb.sub.1 and 50 g of
hydroxysanshool were mixed with 270 g of lactose, 120 g of
microcrystalline cellulose, and 10 g of magnesium stearate, and
this mixture was tableted with a single punch tableting machine to
produce tablets each weighing 250 mg and having a diameter of 9
mm.
[0069] Each of these tablets contains 25 mg each of ginsenoside
Rb.sub.1 and hydroxysanshool. The tablets are to be taken
internally at a dose of 3 to 10 tablets a day in several divided
portions according to symptoms.
[0070] Preparation Example 2
[0071] A 25 g portion of ginsenoside Rg.sub.1 and 25 g of
hydroxysanshool were mixed with 950 g of corn starch, and the
mixture was kneaded by adding water. The resultant was granulated
using a screen with a mesh size of 1 mm.times.1 mm and dried to
obtain a granule preparation.
[0072] A 1 g portion of this granule preparation contains 25 mg
each of ginsenoside Rg.sub.1 and hydroxysanshool. The granule
preparation is to be taken internally at a dose of 2 to 6 g a day
in several divided portions according to symptoms.
[0073] Preparation Example 3
[0074] A 50 g portion of shogaol and 50 g of hydroxysanshool were
mixed with 210 g of lactose, 120 g of starch, 50 g of talc, and 20
g of magnesium stearate, and the mixture was filled into hard
capsules in amounts of 250 mg each to obtain a capsule
preparation.
[0075] Each of these capsules contains 25 mg each of shogaol and
hydroxysanshool. The capsule preparation is to be taken internally
at a dose of 3 to 10 capsules a day in several divided portions
according to symptoms.
[0076] Preparation Example 4
[0077] A 50 g portion of a ginseng extract and 50 g of a Japanese
pepper extract were mixed with 270 g of lactose, 120 g of
microcrystalline cellulose, and 10 g of magnesium stearate, and
this mixture was tableted with a single punch tableting machine to
produce tablets each weighing 250 mg and having a diameter of 9
mm.
[0078] Each of these tablets contains 25 mg each of the ginseng
extract and the Japanese pepper extract. The tablets are to be
taken internally at a dose of 3 to 10 tablets a day in several
divided portions according to symptoms.
[0079] Preparation Example 5
[0080] A 25 g portion of a ginseng extract, 25 g of a Japanese
pepper extract, and 25 g of a dried ginger rhizome extract were
mixed with 925 g of corn starch, and the mixture was kneaded by
adding water. The resultant was granulated using a screen with a
mesh size of 1 mm.times.1 mm and dried to obtain a granule
preparation.
[0081] A 1 g portion of the present granule preparation contains 25
mg each of the ginseng extract, Japanese pepper extract, and dried
ginger rhizome extract. The granule preparation is to be taken
internally at a dose of 2 to 6 g a day in several divided portions
according to symptoms.
[0082] Preparation Example 6
[0083] Production Of Daikenchutou Extract Granules:
[0084] Crude drugs were cut out, and 3 kg of ginseng, 2 kg of
Japanese pepper, and 5 kg of dried ginger rhizome were weighed and
combined. Purified water was added thereto in an approximately
12-fold amount, and the temperature was raised to 95 to 100.degree.
C. while stirring. Extraction was then performed for about 60
minutes. After completion of the extraction, the extract was
subjected to solid-liquid separation, and the separated liquid was
concentrated under reduced pressure. Subsequently, sucrose fatty
acid ester was added and mixed in an amount corresponding to 1.0%
of the solid fraction, and the mixture was spray-dried to obtain
1.25 kg of dry extract. Then 10 kg of Saccharum granorum (powdered
syrup) , 3.7125 kg of lactose, and 0. 0375 kg of magnesium stearate
were added to 1.25 kg of the dried extract and mixed. This mixture
was tableted with a tableting machine according to a conventional
method, and the tablets were then subjected to pulverization,
granulation, and sieving to obtain a satisfactory granule
preparation.
[0085] A 15 g portion of the present granule preparation contains
1.25 g of a crude drug extract and 10 g of powdered syrup. The
granule preparation is to be taken internally at a dose of 15 g a
day in two to three divided portions according to symptoms.
INDUSTRIAL APPLICABILITY
[0086] The adrenomedullin production-enhancing agent of the
invention effectively promotes the production of adrenomedullin,
which has various physiological effects including increasing blood
flow as well as anti-inflammatory effects. The sustainability of
its effects and its safety are excellent. The adrenomedullin
production-enhancing agent of the invention can thus be used in the
treatment of various diseases, and is effective in the prevention
and treatment of diseases including, for example, inflammatory
bowel diseases such as Crohn's disease, myocardial disorders,
pulmonary hypertension, bone disorders, myometrial contraction,
urinary disorders, non-bacterial inflammatory disorders, and
hepatitis.
BRIEF DESCRIPTION OF THE DRAWINGS
[0087] FIG. 1 is a diagram showing the results of measurement of
adrenomedullin concentration in plasma of portal vein blood
obtained as described in Example 1. Furthermore, * indicates a risk
rate of 5% or less, while ** indicates a risk rate of 1% or
less.
[0088] FIG. 2 is a diagram showing the rates of increase in VC as a
result of administration of Daikenchutou in Example 2.
[0089] FIG. 3 is a diagram showing the rates of increase in VC as a
result of administration of ginseng in Example 2.
[0090] FIG. 4 is a diagram showing the rates of increase in VC as a
result of administration of dried ginger rhizome in Example 2.
[0091] FIG. 5 is a diagram showing the rates of increase in VC as a
result of administration of Japanese pepper in Example 2.
[0092] FIG. 6 is a diagram showing the rates of increase in VC as a
result of administration of dried ginger rhizome and ginseng in
Example 2.
[0093] FIG. 7 is a diagram showing the rates of increase in VC as a
result of administration of ginseng and Japanese pepper in Example
2.
[0094] FIG. 8 is a diagram showing the rates of increase in VC as a
result of administration of dried ginger rhizome and Japanese
pepper in Example 2.
[0095] FIG. 9 is a diagram showing the rates of increase in VC in
the ADM antagonist-pretreated group and the Daikenchutou-treated
group in Example 3.
[0096] FIG. 10 is a diagram showing the rates of increase in VC as
a result of administration of hydroxysanshool alone and combined
administration of hydroxysanshool and a ginseng extract in Example
4.
[0097] FIG. 11 is a diagram showing the rates of increase in VC as
a result of administration of ginsenoside Rb.sub.1 alone and
combined administration of hydroxysanshool and ginsenoside Rb.sub.1
in Example 4.
[0098] FIG. 12 is a diagram showing the rates of increase in VC as
a result of administration of ginsenoside Rg.sub.1 alone and
combined administration of hydroxysanshool and ginsenoside Rg.sub.1
in Example 4.
[0099] FIG. 13 is a diagram showing the rates of increase in VC as
a result of administration of 6-shogaol alone and combined
administration of hydroxysanshool and 6-shogaol in Example 5.
[0100] FIG. 14 is a diagram showing the results of measurement of
necrotic area in Example 6.
[0101] FIG. 15 is a diagram showing the results of measurement of
frequency of adhesions in Example 6.
[0102] FIG. 16 is a photograph of the large intestine three days
after induction of enteritis in Example 6.
[0103] FIG. 17 is a diagram showing the results of measurement of
hepatic fibrosis in Example 7.
[0104] FIG. 18 is a diagram showing the results of quantification
of the amount of hyaluronic acid in blood in Example 7.
[0105] FIG. 19 is a diagram showing the results of quantification
of the amount of hydroxyproline in liver in Example 7.
* * * * *