U.S. patent application number 10/572895 was filed with the patent office on 2007-05-10 for method of inhibiting alveolar bone resorption and periodontal membrane loss and composition for internal use to be used therein.
Invention is credited to Hidehiko Otsuki, Yukiyo Sekimoto, Akane Takemura.
Application Number | 20070104802 10/572895 |
Document ID | / |
Family ID | 34587149 |
Filed Date | 2007-05-10 |
United States Patent
Application |
20070104802 |
Kind Code |
A1 |
Sekimoto; Yukiyo ; et
al. |
May 10, 2007 |
Method of inhibiting alveolar bone resorption and periodontal
membrane loss and composition for internal use to be used
therein
Abstract
Alveolar bone resorption, periodontal membrane loss, and
gingival recession can be inhibited by administering 500 mg to 2000
mg per day of calcium, 10 mg to 40 mg per day of soy isoflavone
aglycone, and vitamin D.sub.3 to persons having a tendency for
decreased bone density, postmeopausal women, and periodontal
disease patients in a maintenance phase; and even an extended
period of administration provides a high degree of safety.
Inventors: |
Sekimoto; Yukiyo;
(Takatsuki-shi, JP) ; Otsuki; Hidehiko;
(Takatsuki-shi, JP) ; Takemura; Akane;
(Takatsuki-shi, JP) |
Correspondence
Address: |
KNOBBE MARTENS OLSON & BEAR LLP
2040 MAIN STREET
FOURTEENTH FLOOR
IRVINE
CA
92614
US
|
Family ID: |
34587149 |
Appl. No.: |
10/572895 |
Filed: |
September 17, 2004 |
PCT Filed: |
September 17, 2004 |
PCT NO: |
PCT/JP04/14123 |
371 Date: |
March 20, 2006 |
Current U.S.
Class: |
424/682 ;
514/167; 514/456 |
Current CPC
Class: |
A61P 1/02 20180101; A61P
19/00 20180101; A61K 31/352 20130101; A61K 31/593 20130101; A61K
33/06 20130101; A61K 36/48 20130101; A61K 31/352 20130101; A61K
2300/00 20130101; A61K 31/593 20130101; A61K 2300/00 20130101; A61K
33/06 20130101; A61K 2300/00 20130101; A61K 36/48 20130101; A61K
2300/00 20130101 |
Class at
Publication: |
424/682 ;
514/167; 514/456 |
International
Class: |
A61K 33/06 20060101
A61K033/06; A61K 31/59 20060101 A61K031/59; A61K 31/353 20060101
A61K031/353 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 19, 2002 |
JP |
2003-32822 |
Claims
1. An oral composition for alveolar bone resorption inhibition and
periodontal membrane loss inhibition, comprising a soy isoflavone
aglycone, calcium, and vitamin D.sub.3.
2. An agent for preventing or treating gingival recession,
comprising a soy isoflavone aglycone, calcium, and vitamin
D.sub.3.
3. An agent for preventing or treating alveolar bone resorption and
periodontal membrane loss, comprising a soy isoflavone aglycone,
calcium, and vitamin D.sub.3.
4. A composition or agent according to claim 1, wherein the
proportion of soy isoflavone aglycone in the composition or agent
is 0.001% to 10% by weight; and the proportion of calcium in the
composition or agent is 0.01% to 50% by weight.
5. A composition or agent according to claim 1, wherein the
composition or agent is for persons having decreased bone density,
postmenopausal women, or periodontal disease patients in a
maintenance phase.
6. A composition according to claim 1, wherein the soy isoflavone
aglycone is an extract from whole-grain soy; the genistein/daidzein
weight ratio in the soy isoflavone aglycone is in the range of 1/1
to 1.5/1; and the proportion of the total weight of genistein and
daidzein in the soy isoflavone aglycone is at least 90%.
7. A method for inhibiting alveolar bone resorption and periodontal
membrane loss, comprising orally administering a composition
according to claim 1.
8. A method for preventing or treating gingival recession,
comprising orally administering a soy isoflavone aglycone, calcium,
and vitamin D.sub.3.
9. A method for preventing or treating alveolar bone resorption and
periodontal membrane loss, comprising orally administering a soy
isoflavone aglycone, calcium, and vitamin D.sub.3.
10. A method according to claim 8, wherein the soy isoflavone
aglycone, calcium, and vitamin D.sub.3 are administered to persons
having decreased bone density, postmenopausal women, or periodontal
disease patients in a maintenance phase.
11. A method according to claim 9, wherein the soy isoflavone
aglycone is administered in an amount of 10 mg to 40 mg per day;
and calcium is administered in an amount of 500 mg to 2000 mg per
day.
12. A composition or agent according to claim 3, wherein the
proportion of soy isoflavone aglycone in the composition or agent
is 0.001% to 10% by weight; and the proportion of calcium in the
composition or agent is 0.01% to 50% by weight.
13. A composition or agent according to claim 3, wherein the
composition or agent is for persons having decreased bone density,
postmenopausal women, or periodontal disease patients in a
maintenance phase.
14. A method for inhibiting alveolar bone resorption and
periodontal membrane loss, comprising orally administering a
composition according to claim 6.
15. A method according to claim 9, wherein the soy isoflavone
aglycone, calcium, and vitamin D.sub.3 are administered to persons
having decreased bone density, postmenopausal women, or periodontal
disease patients in a maintenance phase.
16. A method according to claim 10, wherein the soy isoflavone
aglycone is administered in an amount of 10 mg to 40 mg per day;
and calcium is administered in an amount of 500 mg to 2000 mg per
day.
Description
TECHNICAL FIELD
[0001] The present invention relates to a method for inhibiting
alveolar bone resorption and periodontal membrane loss of persons
having a tendency for decreased bone density, postmenopausal women,
and periodontal disease patients in a maintenance phase; a method
for inhibiting gingival recession of such persons; and an oral
composition, an agent for preventing or treating gingival
recession, and an agent for preventing or treating alveolar bone
resorption and periodontal membrane loss, each comprising a soy
isoflavone aglycone, calcium, and vitamin D.sub.3.
BACKGROUND ART
[0002] Decreased bone density is caused by aging, nutrition/diet
problems such as shortage of calcium intake, lack of exercise,
medicines such as adrenal corticosteroids, etc. In particular,
women tend to suffer a rapid decrease of bone mass because of
decreased estrogen secretion due to menopause. Here, a person
having decreased bone density is one whose bone mineral density is
1 SD (Standard Deviation) or more below the young adult mean
(>-1 SD), according to the criteria proposed by the WHO research
group in 1994; or one whose bone density is 80% or less of the
young adult mean (YAM), according to the criteria proposed by the
Japanese Society for Bone and Mineral Research in 1996.
[0003] Periodontal disease is an infection caused by specific
periodontal disease-causing bacteria. Examples of such periodontal
disease-causing bacteria include gram-negative anaerobic bacteria
such as Porphyromonas gingivalis. Periodontal disease-causing
bacteria increase in the plaque that forms around the root of a
tooth cervix, inducing chronic inflammation of the surrounding
tissues including gingiva, periodontal membrane, and alveolar bone,
and thus developing symptoms of periodontal disease. Because of
chronic inflammation induced by periodontal disease-causing
bacteria, periodontal disease advances resorption of alveolar bone,
which supports the teeth, and it is the greatest cause of lost
teeth in developed countries.
[0004] With respect to postmenopausal women, it has been revealed
that there is a correlation between the drop in bone density and
the progression of periodontal disease (Yasunari Kurosu et al.,
Nihon Shika Hozongaku Zassi (The Japanese Journal of Conservative
Dentistry), 1998), and decreased bone mass is regarded as a risk
factor for periodontal disease.
[0005] The finding that a drop in bone density relates to the
progression of periodontal disease led to an animal experiment
using bisphosphonate, which is a bone resorption inhibitor. It was
thus reported that bisphosphonate is useful for inhibiting alveolar
bone resorption in experimental periodontitis (Reddy et al., J
Periodontol, 66 (3), 211-217, 1995). However, since bisphosphonate
has strong side effects, it is not suitable for periodontal disease
treatment, which requires long-term medication. Therefore, the
development of a pharmaceutical preparation having a high degree of
safety has been desired.
[0006] Since periodontal tissues do not recover completely even
after periodontal disease treatment, the dental root surface of a
treated area is exposed. Because inflamed parts and affected
tissues are physically removed in the process of treatment, the
treated area tends to suffer gingival recession. Accordingly, in a
maintenance phase, when the symptoms are stable after periodontal
disease treatment, gingival recession and a relapse of periodontal
disease are highly likely to occur. Therefore, the development of a
pharmaceutical preparation that has a high degree of safety and
that is useful for inhibiting gingival recession and maintaining
periodontal tissues such as alveolar bone and periodontal membrane
is desired.
[0007] It is well known that calcium is a necessary nutritional
component for growing children, pregnant women, etc. to maintain or
enhance the bone calcium density of alveolar bone ("X-ray, optical
microscopic, and scanning electron microscopic research on alveolar
bone changes due to vitamin D deficiency", The Journal of the
Kyushu Dental Society, 46, 67-88, 1992). Accordingly, there are a
large number of calcium food supplements available. It is known
that specific calcium salts (such as calcium pantothenate) inhibit
alveolar bone resorption, which is related to periodontal disease
(Japanese Unexamined Patent Publication No. 1996-133969). It is
thus accepted that intake of calcium and calcium salts is effective
to some extent in the prevention of periodontal disease.
[0008] In contrast, it is known that soy isoflavones abundantly
contain components having antibacterial properties and natural bone
metabolism improvement actions. Therefore, soy isoflavones have
been used together with calcium salts and vitamin D in oral
nutritional supplements for osteoporosis prevention and whole-body
bone strengthening, and, in the United States of America, they have
been sold under the trade names of "Osteo Soy" (FreeLife, U.S.A.)
and "Soylife" (Schouten, U.S.A.). Like ordinary calcium
supplements, these are in oral tablet, capsule, and powder forms
and are not intended for the prevention of periodontal disease.
Among soy isoflavones, genistein shows antibacterial activity
against periodontal disease-causing bacteria; therefore, it has
been used as an antiperiodontitis agent (Japanese Unexamined Patent
Publication No. 1992-283518) and has been used together with
high-solubility calcium in a food composition for the prevention of
periodontal disease or the prevention of periodontal disease
progression (Japanese Unexamined Patent Publication No.
1999-243910). However, the above periodontal disease prevention is
based on the antibacterial properties of soy isoflavones, and there
has been no reported case of inhibited periodontal membrane
loss.
DISCLOSURE OF THE INVENTION
[0009] An object of the present invention is to provide a method,
oral composition, and prevention or treatment agent for inhibiting
alveolar bone resorption and periodontal membrane loss or
inhibiting gingival recession in persons having a tendency for
decreased bone density and postmenopausal women, while having a
high degree of safety even when the composition or agent is taken
for extended periods of time.
[0010] The present inventors conducted extensive research and, as a
result, found that alveolar bone resorption, periodontal membrane
loss, and gingival recession can be inhibited by the combined use
of a soy isoflavone aglycone, calcium, and vitamin D.sub.3. Based
on this finding, the inventors have accomplished the present
invention.
[0011] Specifically, the present invention provides the following
oral compositions, prevention or treatment agents, and methods.
[0012] 1. An oral composition for alveolar bone resorption
inhibition and periodontal membrane loss inhibition, comprising a
soy isoflavone aglycone, calcium, and vitamin D.sub.3.
[0013] 2. An agent for preventing or treating gingival recession,
comprising a soy isoflavone aglycone, calcium, and vitamin
D.sub.3.
[0014] 3. An agent for preventing or treating alveolar bone
resorption and periodontal membrane loss, comprising a soy
isoflavone aglycone, calcium, and vitamin D.sub.3.
[0015] 4. A composition or agent according to any one of items 1 to
3, wherein the proportion of soy isoflavone aglycone in the
composition or agent is 0.001% to 10% by weight; and the proportion
of calcium in the composition or agent is 0.01% to 50% by
weight.
[0016] 5. A composition or agent according to any one of items 1 to
3, wherein the composition or agent is for persons having decreased
bone density, postmenopausal women, or periodontal disease patients
in a maintenance phase.
[0017] 6. A composition according to item 1, wherein the soy
isoflavone aglycone is an extract from whole-grain soy; the
genistein/daidzein weight ratio in the soy isoflavone aglycone is
in the range of 1/1 to 1.5/1; and the proportion of the total
weight of genistein and daidzein in the soy isoflavone aglycone is
at least 90%.
[0018] 7. A method for inhibiting alveolar bone resorption and
periodontal membrane loss, comprising orally administering a
composition according to any one of items 1 to 6.
[0019] 8. A method for preventing or treating gingival recession,
comprising orally administering a soy isoflavone aglycone, calcium,
and vitamin D.sub.3.
[0020] 9. A method for preventing or treating alveolar bone
resorption and periodontal membrane loss, comprising orally
administering a soy isoflavone aglycone, calcium, and vitamin
D.sub.3.
[0021] 10. A method according to item 8 or 9, wherein the soy
isoflavone aglycone, calcium, and vitamin D.sub.3 are administered
to persons having decreased bone density, postmenopausal women, or
periodontal disease patients in a maintenance phase. 11. A method
according to item 9 or 10, wherein the soy
[0022] isoflavone aglycone is administered in an amount of 10 mg to
40 mg per day; and calcium is administered in an amount of 500 mg
to 2000 mg per day.
[0023] In the present specification, gingival recession includes a
gingival margin receding down from the boundary between the tooth
cementum and enamel, i.e., the cement/enamel junction, toward the
root apex, thereby exposing a dental root. Gingival recession can
be quantified by, for example, the numerical value obtained by
subtracting the periodontal pocket depth (PD) from the clinical
attachment level (CAL) (see FIG. 1). In the present specification,
periodontal disease patients in a maintenance phase encompass those
who, having finished periodontal disease treatment, are in a
clinically recovered and stable condition but need observation.
[0024] Soy isoflavone aglycones in the present invention are soy
isoflavone nonglycosides such as genistein, daidzein, glycitein,
etc., and can be usually obtained as soy isoflavone glycoside
hydrolysates. Such soy isoflavone aglycones can be obtained by, for
example, extracting a glycoside from seeds (whole-grain soybeans)
of Glycine max Merrill (Leguminosae family) according to such a
known method as disclosed in Japanese Unexamined Patent Publication
No. 1987-126186, and subjecting the obtained glycoside to acid
heating or .beta.-glucuronidase enzyme hydrolysis in a purification
step.
[0025] There are no limitations on the methods for obtaining
soybean extracts from whole-grain soybeans or isoflavone glycosides
from ground soybeans, and obtaining aglycones from the glycosides.
If soybean hypocotyls are used instead of whole-grain soybeans,
isoflavones can be efficiently obtained; however, the content of
glycosides such as daidzin, glycitin, etc., is high, and such
glycosides are not easily converted into nonglycosides such as
genistein, daidzein, etc. by hydrolysis. Therefore, whole-grain
soybeans are more advantageous than soybean hypocotyls as starting
materials for soy isoflavone aglycones such as genistein, daidzein,
etc.
[0026] The soy isoflavone aglycone used in the present invention is
preferably at least one aglycone selected from the group consisting
of genistein and daidzein. It is preferable to use genistein and
daidzein in combination such that the genistein/daidzein weight
ratio is in the range of 1/1 to 1.5/1. In the soy isoflavone
aglycone, the content of genistein is preferably greater than that
of daidzein. It is also preferable that the proportion of the total
weight of genistein and daidzein in the soy isoflavone aglycone is
at least 90% by weight. Soy isoflavone aglycones may be powdered
with excipients added thereto to be in forms suitable for various
oral preparations or may be used in forms suitable for beverages,
such as preparations with emulsifiers and solubilizers added
thereto, cyclodextrin inclusion preparations, etc.
[0027] The proportion of soy isoflavone aglycone in the composition
or the prevention/treatment agent of the present invention is not
limited as long as it allows the objects of the invention to be
achieved. The proportion of soy isoflavone aglycone in the
composition or the prevention/treatment agent of the invention is
usually 0.001% to 10% by weight, and preferably 0.005% to 5.0% by
weight. In the method for inhibition of alveolar bone resorption
and periodontal membrane loss and the method for prevention or
treatment thereof according to the present invention, soy
isoflavone aglycone is usually administered to an adult in an
amount of 10 mg to 40 mg per day, and preferably in an amount of 10
mg to 20 mg per day; and the administration of soy isoflavone
aglycone may be carried out in one dose a day or in two or more
divided doses a day.
[0028] Any natural or synthetic calcium may be used in the present
invention. Examples of natural forms of calcium include those
derived from oyster shells, egg shells, corals, cattle bones, milk,
etc.; and these items themselves and crude products of these such
as ground products, dried products, etc, can be used as calcium
sources. Examples of synthetic forms of calcium include calcium
gluconate, calcium lactate, calcium chloride, calcium
glycerophosphate, calcium pantothenate, calcium tertiary phosphate,
calcium carbonate, calcium citrate, etc. Such forms of calcium may
be used singly or in combination of two or more.
[0029] The proportion of calcium in the composition or the
prevention/treatment agent of the present invention is not limited
as long as it allows the objects of the invention to be achieved.
The proportion of calcium in the composition or the
prevention/treatment agent of the invention is usually 0.01% to 50%
by weight, and preferably 0.1% to 20% by weight, calculated as the
Ca content. In the method for inhibition of alveolar bone
resorption and periodontal membrane loss and the method for
prevention or treatment thereof according to the present invention,
calcium is usually administered to an adult in an amount of 500 mg
to 2000 mg per day, and preferably in an amount of 500 mg to 1000
mg per day; and the administration of calcium may be carried out in
one dose a day or in two or more divided doses a day.
[0030] Vitamin D.sub.3, which is used in the present invention, is
also called cholecalciferol. When used in the composition or the
prevention/treatment agent of the invention, vitamin D.sub.3 may be
in crystalline form or may be in the form of a preparation for
stabilization. The proportion of vitamin D.sub.3 in the composition
or the prevention/treatment agent of the present invention is not
limited as long as it allows the objects of the invention to be
achieved. The proportion of vitamin D.sub.3 in the composition or
the prevention/treatment agent of the invention is usually
10.sup.-8% to 10.sup.-1% by weight, and preferably 10.sup.-7% to
10.sup.-2% by weight. In the method for inhibition of alveolar bone
resorption and periodontal membrane loss and the method for
prevention or treatment thereof according to the present invention,
vitamin D.sub.3 is usually administered to an adult in an amount of
200 IU to 800 IU per day, and preferably in an amount of 200 IU to
400 IU per day; and the administration of vitamin D.sub.3 can be
carried out in one dose a day or in two or more divided doses a
day.
[0031] The composition of the present invention is an oral
composition for alveolar bone resorption inhibition and periodontal
membrane loss inhibition. Examples of oral compositions include
food compositions, pharmaceutical compositions, etc.; and
preferable examples thereof are food compositions. The forms of
oral compositions of the present invention encompass those used in
the fields of foods and pharmaceuticals; however, preferable forms
are those which allow the composition to stay in the oral cavity
for a long time, such as troches, chewing gums, chewable tablets,
gummy candies, candies, etc. Among such forms, troches and chewable
tablets are especially preferable. Tablets such as troches and
chewable tablets are preferably produced by the following
method.
[0032] A powder mixture (powder particle diameter: no more than 250
.mu.m) of soy isoflavone aglycone with other components such as
calcium, sugar alcohol, and excipient are wetted by alcohol, water,
and optionally a disaccharide-or-higher polymerized reducing sugar
such as polydextrose, reduction dextrin, maltitol, maltotriitol,
etc. or a combination of such disaccharide-or-higher polymerized
reducing sugars, and granulated. The resulting granules are mixed
with flavor, vitamin D.sub.3 preparation, lubricant, useful
components that are sensitive to heat or moisture, etc. as needed,
and tableted. The resulting tablets were then formed into tablets
having a diameter of 12 mm to 20 mm, a thickness of 3 mm to 6 mm,
and a weight of 500 mg to 3000 mg per tablet, using a rotary
tableting machine. The tablet hardness as measured in the diameter
direction using a Kiya hardness tester is preferably within the
range of 5 kg to 15 kg, and most preferably within the range of 7
kg to 13 kg.
[0033] The prevention/treatment agent of the present invention is
useful for preventing or treating alveolar bone resorption and
periodontal membrane loss or for preventing or treating gingival
recession, and the agent comprises a soy isoflavone aglycone,
calcium, and vitamin D.sub.3 as effective components. The form of
the prevention/treatment agent of the invention is not limited as
long as it is suitable for oral ingestion, and the
prevention/treatment agent may be, for example, in forms that can
be used for oral ingestion in foods, pharmaceuticals, etc.
[0034] The oral composition and the prevention/treatment agent of
the present invention may contain additives typically used in
foods, such as pH adjusters, organic acids, sugar alcohols,
sweeteners, flavors, dental plaque formation inhibiting materials,
and bad breath inhibiting materials; edible additives used in
pharmaceuticals, such as excipients, other effective components,
and carriers; etc. as needed, as long as the effects of the present
invention are not impaired thereby. Examples of materials for such
additives include phosphates, malic acid, citric acid, lactic acid,
pantothenic acid, sugar alchols such as sorbitol, xylitol,
erythritol, palatinit, palatinose, maltitol, and reducing starch
sugars, isomerized sugars such as reducing
isomaltooligosaccharides, gum bases, gum arabic, gelatin, cetyl
methyl cellulose, sodium saccharin, aspartame, magnesium stearate,
granulated sugar, powdered sugar, starch syrup, microcrystalline
cellulose, 1-menthol, D-group vitamins other than vitamin D.sub.3,
K-group vitamins such as vitamin K, vitamin P, lysine, magnesium
salts, calcitonin, ipriflavone, etc. The amounts of such additives
are not limited and can be suitably determined in accordance with
the purpose of use, etc.
[0035] The method for inhibiting alveolar bone resorption and
periodontal membrane loss according to the present invention
comprises orally administering at least one member selected from
the group consisting of the above oral compositions and
prevention/treatment agents. The method for preventing or treating
gingival recession and the method for preventing or treating
alveolar bone resorption and periodontal membrane loss according to
the present invention comprise orally administering a soy
isoflavone aglycone, calcium, and vitamin D.sub.3.
[0036] In these methods, the subject for administration is not
limited as long as it is a mammal; however, preferable subjects for
administration are persons having decreased bone density,
postmenopausal women, and periodontal disease patients in a
maintenance phase. A person having decreased bone density is one
whose bone mineral density is 1 SD (Standard Deviation) or more
below the young adult mean (>-1 SD).
[0037] The administration amount, form, etc. of the oral
composition and the prevention/treatment agent of the present
invention are described above.
BRIEF DESCRIPTION OF THE DRAWINGS
[0038] FIG. 1 is a sectional view of a tooth crown, dental root,
and periodontal tissue, showing the relationship between CAL, PD,
and gingival recession.
[0039] FIG. 2 is a graph showing the change of MMP-8 level with
time in the gingival crevicular fluid (GCF) obtained in Test
Example 1.
BEST MODE FOR CARRYING OUT THE INVENTION
[0040] The present invention is described below in more detail with
reference to Examples and Test Examples. However, the present
invention is not limited to these examples. In these examples,
percentages are by weight unless otherwise indicated.
TEST EXAMPLE 1
Test for Bone Resorption Inhibition and Periodontal Membrane Loss
Inhibition Using Periodontal Maintenance Patients
1. Test Subjects
[0041] The test subjects were 50 women periodontal disease patients
at least one year after menopause. In selecting the test subjects,
it was confirmed that they did not have any critical systemic
underlying disease, were not under hormone replacement therapy or
osteoporosis medicine treatment, and, within the past three months,
had not undergone surgical periodontal treatment or antibiotic
medication. The 50 test subjects were randomly assigned to two
groups: a test group and a placebo group.
2. Test Pharmaceutical Preparation
[0042] Prescription of Test Pharmaceutical Preparation:
TABLE-US-00001 Non-calcinated shell calcium 650 mg (calcium
content: 250 mg) Soy isoflavone extract 17 mg (containing 5 mg of
soy isoflavone aglycon) (genistein:daidzein = 1.3:1) Vitamin
D.sub.3 2.5 .mu.g (100 IU) Sugar alcohol Balance Sweetener, Flavor,
etc. 140 mg Total 2000 mg/tablet
3. Test Method
[0043] The subjects ingested two tablets of the test pharmaceutical
preparation after breakfast for 24 weeks. At the time of tablet
distribution (hereinafter referred to as baseline), and in the 12th
and 24th weeks after the start of tablet ingestion, they were
subjected to inquiries, standard X-ray photography of the oral
cavity, periodontal tissue examination, and sampling of gingival
crevicular fluid (hereinafter referred to as GCF) for biochemical
marker measurement. The amounts of Ca and soy isoflavone daidzein
and genistein that the subjects ingested from foods, etc. were
calculated from meal records by registered dietitians.
4. Periodontal Tissue Examination
[0044] 1) Periodontal Pocket Depth (hereinafter referred to as PD)
and clinical attachment level (hereinafter referred to as CAL)
[0045] PD and CAL of all the teeth except the third molars were
measured in units of 0.5 mm by the six-point method, using a 15 mm
probe (UNC15, Hu-Friedy, USA) with a scale graduated in 1 mm
increments. CAL is the distance from the cement/enamel junction to
the periodontal pocket bottom (see FIG. 1). [0046] 2) Gingival
Recession
[0047] Gingival recession was indicated by the numerical value
obtained by subtracting the PD from the CAL at each point. [0048]
3) Alveolar Bone Height (Hereinafter Referred to as ACH)
[0049] Standard X-ray photography was performed on the molar sites
on both sides by the bite wing method, and the distance from the
cement/enamel junction to the alveolar bone crest along the dental
root surface was measured by the same skilled person. 5.
Measurement of Biochemical Marker in GCF
[0050] GCF was sampled from mesiobuccal sites of the third molars
on the upper jaw right side and the lower jaw left side, using
periopaper (Proflow.TM. Incorporated, USA). Periopaper was inserted
to the bottom of a pocket, left for 10 seconds, taken out to
measure the amount of GCF by a Periotron 8000 (Harco Electronics,
USA). After periopaper was placed into a microtube, it was used to
measure matrix metalloprotease-8 (hereinafter referred to as
MMP-8), which is an index of connective tissue damage. After
extraction processing with Tris chloride buffer solution (pH 7.6),
MMP-8 was measured using MMP-8 Human Biotrak ELISA System (Amersham
Biosciences, USA). The whole amount of MMP-8 of each GCF sample was
calculated from the measurement results.
6. Statistics Analysis
[0051] The subjects' background factors, ACH, PD, CAL, and gingival
recession were all analyzed by a one-way layout analysis of
variance (ANOVA) and subsequent multiple comparison test. PD and
CAL values were used for analysis, with the figure at the first
decimal place being ignored. Change with time within group was
analyzed by a Bonferroni-correction nonparametric test. For all
analyses, a two-sided significance level of 5% was applied.
[0052] 7. Results [0053] 1) General Condition
[0054] No significant differences were recognized between groups in
terms of background factors: age, BMI, menopause age, the number of
years after menopause, the number of smokers, the number of teeth
present, and the average amounts of ingestion of Ca, daidzein, and
genistein from foods during the test period. [0055] 2) Periodontal
Tissue Examination
[0056] No significant differences were recognized between the
groups in terms of the subject average values of CAL, gingival
recession, and ACH at the time of baseline. [0057] 2-1) CAL
[0058] Table 1 shows CAL average values and average rates of change
(which are obtained by dividing the amount of change in the CAL
average value at 12 or 24 weeks by the baseline CAL average value).
In both groups, areas that had at least 3 mm CAL at the time of
baseline, when they were under maintenance care after treatment,
showed significant CAL decrease from the baseline at the 12th and
24th weeks. In order to adjust for the difference between the
groups at the time of baseline, analysis was conducted on the CAL
average rates of change obtained by dividing the amount of change
in CAL at 12 or 24 weeks by the baseline CAL value. As the result,
in comparison with the placebo group, the test group showed
significant improvement in CAL at the 24th week. TABLE-US-00002
TABLE 1 CAL average value(mm) CAL rate of change(%) Group Baseline
12th week 24th week .DELTA.12 weeks .DELTA.24 weeks Test 3.90 3.42
3.34 -12.5 -14.2 group Placebo 4.08 3.75 3.74 -8.9 -8.9 group
[0059] 2-2) Gingival Recession
[0060] Table 2 shows gingival recession average values and rates of
change (which are obtained by dividing the amount of change in the
gingival recession average value at 12 or 24 weeks by the baseline
gingival recession average value). In the test group and the
placebo group, areas that had gingival recession at the beginning
of the test showed significant improvement in gingival recession at
the 12th and 24th weeks. However, the test group showed significant
difference from the placebo group in the gingival recession rate of
change at the end of the test. TABLE-US-00003 TABLE 2 Gingival
recession Gingival recession average value(mm) rate of change(%)
Group Baseline 12th week 24th week .DELTA.12 weeks .DELTA.24 weeks
Test 1.70 1.19 1.08 -30.1 -37.8 group Placebo 1.97 1.74 1.74 -15.0
-16.4 group
[0061] 2-3) ACH
[0062] Table 3 shows ACH rates of change (which are obtained by
dividing the amount of change in the ACH average value at 12 or 24
weeks by the baseline ACH average value). In order to adjust for
the difference between the groups at the time of baseline, analysis
was conducted on the ACH average rates of change obtained by
dividing the amount of change in ACH at 12 or 24 weeks by the
baseline ACH value. As the result, in comparison with the placebo
group, the test group showed a tendency to inhibit ACH and, in
particular, there was significant difference from the placebo group
at the 12th week. TABLE-US-00004 TABLE 3 ACH rate of change (%)
Group .DELTA.12 weeks .DELTA.24 weeks Test group 0.3 0.3 Placebo
group 7.1 9.5
[0063] 2-4) Level of MMP-8 in GCF
[0064] FIG. 2 shows the MMP-8 average values at the time of
baseline, the 12th week, and the 24th week. The placebo group did
not show any changes, while the test group showed significant
decreases from the baseline at the 12th and 24th weeks.
[0065] The above test results reveal the following points.
[0066] The present test pharmaceutical preparation promoted
improvement in the CAL of postmenopausal periodontal disease
patients in a maintenance phase. It was thus found that the
pharmaceutical preparation has the action of improving connective
tissue attachment by the periodontal membrane, that is, the normal
periodontal tissue attachment obtained by the periodontal membrane
lying between the alveolar bone and dental root surface (cementum).
Since the present test pharmaceutical preparation inhibited the
level of MMP-8, it was found that the pharmaceutical preparation
has the action of preventing loss of connective tissue mainly
composed of periodontal membrane, thus being useful for inhibiting
periodontal membrane loss.
[0067] Although alveolar bone resorption tends to make rapid
progress in postmenopausal women, the present test group showed an
inhibited increase in ACH in comparison with the placebo group. An
effect of inhibiting alveolar bone resorption was thus found.
[0068] Although gingival recession tends to occur easily in treated
areas, it was improved in the present test. It was thus found that
not only a relapse of periodontal disease but also dental root
surface caries and hypersensitivity due to exposed roots can be
prevented and/or improved, and that aesthetic aspects can also be
improved.
Example 1
Granule (Ingestion: 4 g per Day)
[0069] Granules were produced by a standard method, using the
following components: TABLE-US-00005 Component Amount (%) Whey
calcium 50.00 (calcium content: 26%) Soy isoflavone extract 1.00
(aglycon content: 30%) (genistein:daidzein = 1.5:1) Oil-soluble
licorice extract 1.00 Vitamin D.sub.3 (vitamin D.sub.3 content:
0.25%) 0.05 Xylitol 40.00 Palatinit Balance Aspartame 0.10 Gum
arabic 1.00 Flavor 2.50 Total 100.00
Example 2
Intraoral Solution Tablet (Ingestion: 4 g per Day)
[0070] Intraoral solution tablets were produced by a standard
method, using the following components: TABLE-US-00006 Component
Amount (%) Oyster shell calcium 35.00 (calcium content: 39%) Soy
isoflavone extract 0.5 (aglycon content: 70%) (genistein:daidzein =
1.3:1) Palatinit Balance Maltitol 30.00 Vitamin D.sub.3 (vitamin
D.sub.3 content: 0.25%) 0.05 Polydextrose 5.00 Sucrose fatty acid
ester 4.00 Cocoa powder 5.00 Sucralose 1.00 Flavor 2.50 Total
100.00
Example 3
Intraoral Solution Tablet (Ingestion: 4 g per Day)
[0071] Intraoral solution tablets were produced by a standard
method, using the following components: TABLE-US-00007 Component
Amount (%) Calcium tertiary phosphate 35.00 (calcium content:
39.1%) Soy isoflavone extract 1.50 (aglycon content: 33%)
(genistein:daidzein = 1.3:1) Vitamin K 0.01 Vitamin D.sub.3
preparation 0.1 (vitamin D.sub.3 content: 0.25%) Xylitol 31.00
Palatinit Balance Citric acid 0.50 Gum Arabic 1.20 Magnesium
stearate 1.00 Flavor 3.00 Total 100.00
Example 4
Chewing Gum (Ingestion: 20 g per Day; 40% of Soy Isoflavone Eluted;
85% of Calcium Eluted; and 40% of Vitamin D.sub.3 Eluted)
[0072] A chewing gum was produced by a standard method, using the
following components: TABLE-US-00008 Component Amount (%) Calcium
gluconate-calcium lactate amorphous 30.00 material (calcium
content: 10%) Tea extract 0.05 Soy isoflavone extract 0.25 (aglycon
content: 70%) (genistein:daidzein = 1.3:1) Gum base 27.00
Erythritol 10.00 Xylitol 38.00 Vitamin D.sub.3 (vitamin D.sub.3
content: 0.25%) 0.0001 Reducing malt sugar syrup Balance Flavor
5.00 Total 100.00
Example 5
Candy (Ingestion: 30 g per Day)
[0073] A candy was produced by a standard method, using the
following components.
[0074] Oyster shell calcium, xylitol, oil-soluble licorice extract,
tea polyphenol extract, vitamin D.sub.3, and powder flavor were
incorporated in the center of the candy in powdered forms.
TABLE-US-00009 Component Amount (%) Oyster shell calcium 5.00
(calcium content: 39%) Oil-soluble licorice extract 0.05 Soy
isoflavone extract 0.1 (aglycon content: 70%) (genistein:daidzein =
1.3:1) Xylitol 5.00 Tea polyphenol extract 0.05 Palatinit Balance
Maltitol 10.00 Aspartame 0.04 Citric acid 0.50 Oil flavor 0.20
Powder flavor 0.30 Vitamin D.sub.3 (vitamin D.sub.3 content: 0.25%)
0.0001 Total 100.00
Example 6
Troche (Ingestion: 8 g per Day)
[0075] A troche was produced by the standard method, using the
following components: TABLE-US-00010 Component Amount (%) Citrate
calcium 10.00 (calcium content: 21%) Calcium tertiary phosphate
15.00 (calcium content: 39%) Soy isoflavone extract 0.25 (aglycon
content: 70%) (genistein : daidzein = 1:1) Vitamin D.sub.3 (vitamin
D.sub.3 content: 0.25%) 0.05 Palatinit 33.00 Xylitol Balance Sodium
carboxymethylcellulose 1.00 Citric acid 2.00 Malic acid 1.00
Aspartame 0.50 Sucrose fatty acid ester 4.00 Flavor 2.00 Total
100.00
Example 7
Drink Formulation (Ingestion: 100 ml per Day)
[0076] A drink formulation was produced by a standard method, using
the following components: TABLE-US-00011 Component Amount
Fermentation lactic acid (50% aqueous solution) 0.97 g Gluconic
acid (50% aqueous solution) 3.88 g Calcium gluconate-calcium
lactate amorphous 5.00 g material (calcium content: 10%) Soy
isoflavone aglycon-.beta. cyclodextrin inclusion 0.350 g material
(aglycon content: 3%) (genistein:daidzein = 1.5:1) Erythritol 7.00
g Sucralose 0.03 g Stevia 0.01 g Vitamin D.sub.3 (vitamin D.sub.3
content: 0.2%) 0.003 g Purified water Balance Total 100 ml
* * * * *