U.S. patent application number 11/522511 was filed with the patent office on 2007-05-31 for composition comprising the extract of actinidia arguta and related species for the prevention and treatment of allergic disease and non-allergic inflammatory disease.
Invention is credited to Mirim Jin, Bongcheol Kim, Sunyoung Kim, Hwa Jun Lee, Eun Jin Park.
Application Number | 20070122508 11/522511 |
Document ID | / |
Family ID | 31946849 |
Filed Date | 2007-05-31 |
United States Patent
Application |
20070122508 |
Kind Code |
A1 |
Kim; Sunyoung ; et
al. |
May 31, 2007 |
Composition comprising the extract of actinidia arguta and related
species for the prevention and treatment of allergic disease and
non-allergic inflammatory disease
Abstract
The present invention provides a pharmaceutical composition
comprising the extract of hardy kiwifruit as an active ingredient
in an effective amount to treat and prevent allergic disease and
non-allergic inflammatory disease by reducing inflammation action,
by inhibiting histamine release from mast cell, and by increasing
the level of Th1 cytokines, IgG2a in serum and reducing the level
of Th2 cytokines and IgE in serum. The present invention also
provides a use of above extract for the preparation of
pharmaceutical composition. The present invention also provides a
health food or food additives, a cosmetic composition, a feed or
feed additives comprising above extract for prevention or
alleviation of allergic disease and non-allergic inflammatory
disease by reducing inflammation action, by inhibiting histamine
release from mast cell, and by increasing the level of Th1
cytokines, IgG2a in serum, and reducing the level of Th2 cytokines
and IgE in serum.
Inventors: |
Kim; Sunyoung; (GyeongGi-Do,
KR) ; Park; Eun Jin; (Seoul, KR) ; Kim;
Bongcheol; (GyeongGi-Do, KR) ; Jin; Mirim;
(Seoul, KR) ; Lee; Hwa Jun; (GyoongGi-Do,
KR) |
Correspondence
Address: |
STERNE, KESSLER, GOLDSTEIN & FOX P.L.L.C.
1100 NEW YORK AVENUE, N.W.
WASHINGTON
DC
20005
US
|
Family ID: |
31946849 |
Appl. No.: |
11/522511 |
Filed: |
September 18, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10646145 |
Aug 22, 2003 |
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11522511 |
Sep 18, 2006 |
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60405295 |
Aug 23, 2002 |
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Current U.S.
Class: |
424/777 |
Current CPC
Class: |
A61K 36/185 20130101;
A61P 27/14 20180101; A61P 27/02 20180101; A23V 2002/00 20130101;
A61P 37/08 20180101; A61P 17/04 20180101; A61P 11/06 20180101; A61P
1/04 20180101; A61P 17/00 20180101; A61P 13/12 20180101; A61P 11/02
20180101; A61P 43/00 20180101; A61P 1/18 20180101; A23L 33/105
20160801; A61P 29/00 20180101; A61P 1/16 20180101; A23V 2002/00
20130101; A23V 2250/70 20130101; A23V 2250/21 20130101; A23V
2250/72 20130101; A23V 2250/7056 20130101; A23V 2250/705 20130101;
A23V 2250/156 20130101; A23V 2002/00 20130101; A23V 2250/032
20130101; A23V 2250/28 20130101; A23V 2250/21 20130101; A23V
2250/0644 20130101 |
Class at
Publication: |
424/777 |
International
Class: |
A61K 36/185 20060101
A61K036/185 |
Claims
1-37. (canceled)
38. A method to regulate an immune response in a mammal, comprising
administering a hardy kiwifruit preparation to the mammal in an
amount sufficient to regulate an immune response in the mammal.
39. The method of claim 38, wherein the hardy kiwifruit preparation
is provided in an amount sufficient to regulate a Th2 and a Th1
immune response in the mammal.
40. The method of claim 38, wherein the hardy kiwifruit preparation
is provided in an amount sufficient to regulate the amount of an
antibody isotype produced by the mammal selected from the group
consisting of IgE, IgG2a, and IgG1.
41. The method of claim 38, wherein the hardy kiwifruit preparation
is provided in an amount sufficient to decrease the production
and/or levels of at least one Th2 cytokine in the mammal or to
increase the level of at least one Th1 cytokine in the mammal.
42. The method of claim 38, wherein the hardy kiwifruit preparation
is provided in an amount sufficient to decrease the level of
expression of a transcription factor in the mammal.
43. The method of claim 42, wherein the transcription factor is
selected from the group consisting of: GATA-3 and NFATc2.
44. The method of claim 38, wherein the mammal has or is at risk of
developing a condition in which enhancement of a Th1 response
and/or suppression of a Th2 response is desirable.
45. The method of claim 44, wherein the mammal has or is at risk of
developing an allergic disease or non-allergic inflammatory
disease.
46. The method of claim 45, wherein the allergic disease is
selected from the group consisting of: asthma and dermatitis.
47. The method of claim 44, wherein the mammal has or is at risk of
developing a viral infection.
48. The method of claim 44, wherein the mammal has or is at risk of
developing a cancer.
49. The method of claim 38, wherein the hardy kiwifruit is selected
from the group consisting of: Actinidia arguta, Actinidia kolomikta
and Actinidia polygama.
50. The method of claim 38, wherein the hardy kiwifruit is
Actinidia arguta.
51. The method of claim 38, wherein the hardy kiwifruit preparation
is an extract or concentrate prepared from a part of the hardy
kiwifruit selected from the group consisting of: the leaf, the
stem, the root, and any combination thereof.
52. The method of claim 38, wherein the hardy kiwifruit preparation
is selected from the group consisting of: fresh fruit, crushed
fruit, boiled fruit, cooked fruit, pressed fruit, and condensed
fruit.
53. The method of claim 38, wherein the hardy kiwifruit is dried
fruit.
54. The method of claim 38, wherein the hardy kiwifruit preparation
is produced by a method that includes a step of drying the
fruit.
55. The method of claim 38, wherein the hardy kiwifruit preparation
is a hardy kiwifruit juice concentrate.
56. The method of claim 38, wherein the hardy kiwifruit preparation
is produced by extraction of fruit in water having a temperature of
between 0.degree. C and about 80.degree. C.
57. The method of claim 38, wherein the hardy kiwifruit preparation
is produced by extraction of fruit in room temperature water.
58. The method of claim 38, wherein the hardy kiwifruit preparation
is produced by direct extraction of a water soluble concentrate of
hardy kiwifruit with ethyl acetate.
59. The method of claim 38, wherein the hardy kiwifruit preparation
is provided in a composition in an amount of between about 0.01%
and about 95% by weight based on the total weight of the
composition.
60. The method of claim 38, wherein the step of administering
comprises administering the hardy kiwifruit preparation with a
carrier, adjuvant, or diluent to the mammal.
61. The method of claim 38, wherein the step of administering
comprises providing the hardy kiwifruit preparation to the mammal
as a tablet, a powder, a capsule, a liquid, a suspension, a granule
or a syrup.
62. The method of claim 38, wherein the step of administering
comprises providing the hardy kiwifruit preparation to the mammal
in a health food.
63. The method of claim 62, wherein the health food is selected
from the group consisting of: bread, breakfast cereals, processed
cheese, unprocessed cheese, dairy products, carbonated drinks,
teas, processed fish products, fruit-based drinks, vegetable-based
drinks, chewing gum, hard confectionery, frozen dairy products,
processed meat products, chocolate, cookies, candy, licorice, ice
creams, dehydrated foods, cut food products, spices, alcoholic
beverages, noodles, fermented foods, and vegetable-based
drinks.
64. The method of claim 38, wherein the step of administering
comprises applying a cosmetic composition comprising the hardy
kiwifruit preparation to the mammal.
65. The method of claim 64, wherein the cosmetic composition is
provided in a form selected from the group consisting of: lotion,
cream, essence, toner, emulsion, pack, soap, shampoo, rinse,
cleanser, body washing solution, washing solution or treatment.
66. The method of claim 38, wherein the step of administering
comprises providing the hardy kiwifruit preparation to the mammal
in a food additive.
67. The method of claim 38, further comprising administering to the
mammal an agent selected from the group consisting of: organic
acids; small organic compounds; carbohydrates; steroids; herbal
preparations; spices; minerals; sterilizers; seasonings; vitamins;
and electrolytes.
68. The method of claim 38, further comprising administering to the
mammal an agent selected from the group consisting of taurine,
fatty acids and fatty acid esters.
69. The method of claim 68, wherein the fatty acids and fatty acid
esters are selected from the group consisting of: glyceryl
tri-2-ethyl hexanoic acid, cetyl 2-ethyl hexanoic acid, isopropyl
myristic acid, butyl myristic acid, isopropyl palmitic acid, ethyl
stearic acid, octyl palmitic acid, isocetyl isostearic acid, butyl
stearic acid, ethyl linoleic acid, isopropyl linoleic acid, ethyl
oleic acid, isocetyl myristic acid, isostearyl myristic acid,
isostearyl palmitic acid, octyldodecyl myristic acid, isocetyl
isostearic acid, diethyl sebasic acid, isopropyl adipic acid,
isoalkyl neopetanoic acid, glyceryl tri(capryl, capric acid),
trimethylopropane tri-2-ethyl hexanoic acid, trimethylopropane
triisostearic acid, pentaerythritol tetra-2 ethyl hexanoic acid,
cetyl caprylic acid, decyl lauric acid, hexyl lauric acid, decyl
myristic acid, myristyl myristic acid, cetyl myristic acid, stearyl
stearic acid, decyl oleic acid, cetyl licinoleic acid, isostearyl
lauric acid, isotridecyl myristic acid, isocetyl palmitic acid,
octyl stearic acid, isocetyl stearic acid, isodecyl oleic acid,
octyldodecyl oleic acid, octyldodecyl linoleic acid, isopropyl
isostearic acid, cetostearyl 2-ethyl hexanoic acid, stearyl 2-ethyl
hexanoic acid, hexyl isostearic acid, ethylene glycol dioctanoic
acid, ethylene glycol dioleic acid, propylene glycol dicapric acid,
propylene glycol di(capryl, capric acid), propylene glycol
dicaprylic acid, neopentylglycol dicapric acid, neopentylglycol
dioctanoic acid, glyceryl tricaprylic acid, glyceryl triundecylic
acid, glyceryl triisopalmitic acid, glyceryl triisostearic acid,
octyldodecyl neopentanoic acid, isostearyl octanoic acid, octyl
isononanoic acid, hexyldecyl neodecanoic acid, octyldodecyl
neodecanoic acid, isocetyl isostearic acid, isostearyl isostearic
acid, octyldecyl isostearic acid, polyglycerin oleanoic acid ester,
polyglycerin isostearic acid ester, triisocetyl citric acid,
triisoalkyl citric acid, triisooctyl citric acid, lauryl lactic
acid, myristyl lactic acid, cetyl lactic acid, octyldecyl lactic
acid, triethyl citric acid, acetyltriethyl citric acid, acetyl
tributyl citric acid, trioctyl citric acid, diisostearyl maleic
acid, di 2-ethylhexyl hydroxy stearic acid, 2-ethyl hexyl succinic
acid, diisobutyl adipic acid, diisopropyl sebasinic acid, dioctyl
sebacinic acid, cholesteryl stearic acid, cholesteryl isostearic
acid, cholesteryl hydroxy stearic acid, cholesteryl hydroxy stearic
acid, cholesteryl oleic acid, dihydrocholesteryl oleic acid,
pitsteryl isostearic acid, pitsteryl oleic acid, isocetyl
12-stealoyl hydroxy stearic acid, stearyl 12-stealoyl hydroxy
stearic acid, isostearyl 12-stealoyl hydroxy stearic acid.
70. The method of claim 38, further comprising administering to the
mammal a different Actinidia species preparation.
71. The method of claim 70, wherein the different Actinidia species
is selected from the group consisting of: A. chenensis, A.
deliciosa , A. arguta, A. polygama, and A. kolomikta.
72. A composition for regulating an immune response in a mammal,
comprising a hardy kiwifruit preparation and at least one
additional active compound for regulating an immune response in a
mammal.
73. The composition of claim 72, wherein the additional active
compound is for treating or preventing allergic disease in a
mammal.
74. The composition of claim 72, wherein the additional active
compound is selected from a group consisting of steroids.
75. The composition of claim 72, wherein the additional active
compound is selected from the group consisting of: citric acid;
fumaric acid; steroids; linear peptides; carbohydrates; herbal
preparations; spices; minerals; sterilizers; seasonings; vitamins;
and electrolytes.
76. The composition of claim 72, wherein the additional active
compound is selected from the group consisting of taurine, fatty
acids and fatty acid esters.
77. The composition of claim 76, wherein the fatty acids and fatty
acid esters are selected from the group consisting of: glyceryl
tri-2-ethyl hexanoic acid, cetyl 2-ethyl hexanoic acid, isopropyl
myristic acid, butyl myristic acid, isopropyl palmitic acid, ethyl
stearic acid, octyl palmitic acid, isocetyl isostearic acid, butyl
stearic acid, ethyl linoleic acid, isopropyl linoleic acid, ethyl
oleic acid, isocetyl myristic acid, isostearyl myristic acid,
isostearyl palmitic acid, octyldodecyl myristic acid, isocetyl
isostearic acid, diethyl sebasic acid, isopropyl adipic acid,
isoalkyl neopetanoic acid, glyceryl tri(capryl, capric acid),
trimethylopropane tri-2-ethyl hexanoic acid, trimethylopropane
triisostearic acid, pentaerythritol tetra-2 ethyl hexanoic acid,
cetyl caprylic acid, decyl lauric acid, hexyl lauric acid, decyl
myristic acid, myristyl myristic acid, cetyl myristic acid, stearyl
stearic acid, decyl oleic acid, cetyl licinoleic acid, isostearyl
lauric acid, isotridecyl myristic acid, isocetyl palmitic acid,
octyl stearic acid, isocetyl stearic acid, isodecyl oleic acid,
octyldodecyl oleic acid, octyldodecyl linoleic acid, isopropyl
isostearic acid, cetostearyl 2-ethyl hexanoic acid, stearyl 2-ethyl
hexanoic acid, hexyl isostearic acid, ethylene glycol dioctanoic
acid, ethylene glycol dioleic acid, propylene glycol dicapric acid,
propylene glycol di(capryl, capric acid), propylene glycol
dicaprylic acid, neopentylglycol dicapric acid, neopentylglycol
dioctanoic acid, glyceryl tricaprylic acid, glyceryl triundecylic
acid, glyceryl triisopalmitic acid, glyceryl triisostearic acid,
octyldodecyl neopentanoic acid, isostearyl octanoic acid, octyl
isononanoic acid, hexyldecyl neodecanoic acid, octyldodecyl
neodecanoic acid, isocetyl isostearic acid, isostearyl isostearic
acid, octyldecyl isostearic acid, polyglycerin oleanoic acid ester,
polyglycerin isostearic acid ester, triisocetyl citric acid,
triisoalkyl citric acid, triisooctyl citric acid, lauryl lactic
acid, myristyl lactic acid, cetyl lactic acid, octyldecyl lactic
acid, triethyl citric acid, acetyltriethyl citric acid, acetyl
tributyl citric acid, trioctyl citric acid, diisostearyl maleic
acid, di 2-ethylhexyl hydroxy stearic acid, 2-ethyl hexyl succinic
acid, diisobutyl adipic acid, diisopropyl sebasinic acid, dioctyl
sebacinic acid, cholesteryl stearic acid, cholesteryl isostearic
acid, cholesteryl hydroxy stearic acid, cholesteryl hydroxy stearic
acid, cholesteryl oleic acid, dihydrocholesteryl oleic acid,
pitsteryl isostearic acid, pitsteryl oleic acid, isocetyl
12-stealoyl hydroxy stearic acid, stearyl 12-stealoyl hydroxy
stearic acid, isostearyl 12-stealoyl hydroxy stearic acid.
78. The composition of claim 72, wherein the composition is
selected from the group consisting of: a pharmaceutical
composition, a health food, a food additive, and a cosmetic.
79. The composition of claim 72, wherein the hardy kiwifruit is
selected from the group consisting of: Actinidia arguta, Actinidia
kolomikta and Actinidia polygama.
80. The composition of claim 72, wherein the hardy kiwifruit is
Actinidia arguta.
81. The composition of claim 72, wherein the hardy kiwifruit
preparation is an extract or concentrate prepared from a part of
the hardy kiwifruit selected from the group consisting of: the
fruit, the leaf, the stem, the root, and any combination
thereof.
82. The composition of claim 72, wherein the hardy kiwifruit is
selected from the group consisting of: fresh fruit, crushed fruit,
boiled fruit, cooked fruit, pressed fruit, and condensed fruit.
83. The composition of claim 72, wherein the hardy kiwifruit is
dried fruit.
84. The composition of claim 72, wherein the hardy kiwifruit
preparation is produced by a method that includes a step of drying
the fruit.
85. The composition of claim 72, wherein the hardy kiwifruit
preparation is a hardy kiwifruit juice concentrate.
86. The composition of claim 72, wherein the hardy kiwifruit
preparation is produced by extraction of fruit in water having a
temperature of between 0.degree. C. and about 80.degree. C.
87. The composition of claim 72, wherein the hardy kiwifruit
preparation is produced by extraction of fruit in room temperature
water.
88. The composition of claim 72, wherein the hardy kiwifruit
preparation is produced by direct extraction of a water soluble
concentrate of hardy kiwifruit with ethyl acetate.
89. The composition of claim 72, wherein the extract is prepared by
extraction of hardy kiwifruit in distilled water.
90. The composition of claim 72, wherein the extract is an ethyl
acetate extract of the hardy kiwifruit.
91. A method to treat a disease or condition that is associated
with dysregulation of immune function, comprising administering to
a mammal a combination of hardy kiwifruit or a preparation thereof
and an agent selected from the group consisting of: steroids; fatty
acids and fatty acid esters.
92. The method of claim 91, wherein the fatty acids and fatty acid
esters are selected from the group consisting of: glyceryl
tri-2-ethyl hexanoic acid, cetyl 2-ethyl hexanoic acid, isopropyl
myristic acid, butyl myristic acid, isopropyl palmitic acid, ethyl
stearic acid, octyl palmitic acid, isocetyl isostearic acid, butyl
stearic acid, ethyl linoleic acid, isopropyl linoleic acid, ethyl
oleic acid, isocetyl myristic acid, isostearyl myristic acid,
isostearyl palmitic acid, octyldodecyl myristic acid, isocetyl
isostearic acid, diethyl sebasic acid, isopropyl adipic acid,
isoalkyl neopetanoic acid, glyceryl tri(capryl, capric acid),
trimethylopropane tri-2-ethyl hexanoic acid, trimethylopropane
triisostearic acid, pentaerythritol tetra-2 ethyl hexanoic acid,
cetyl caprylic acid, decyl lauric acid, hexyl lauric acid, decyl
myristic acid, myristyl myristic acid, cetyl myristic acid, stearyl
stearic acid, decyl oleic acid, cetyl licinoleic acid, isostearyl
lauric acid, isotridecyl myristic acid, isocetyl palmitic acid,
octyl stearic acid, isocetyl stearic acid, isodecyl oleic acid,
octyldodecyl oleic acid, octyldodecyl linoleic acid, isopropyl
isostearic acid, cetostearyl 2-ethyl hexanoic acid, stearyl 2-ethyl
hexanoic acid, hexyl isostearic acid, ethylene glycol dioctanoic
acid, ethylene glycol dioleic acid, propylene glycol dicapric acid,
propylene glycol di(capryl, capric acid), propylene glycol
dicaprylic acid, neopentylglycol dicapric acid, neopentylglycol
dioctanoic acid, glyceryl tricaprylic acid, glyceryl triundecylic
acid, glyceryl triisopalmitic acid, glyceryl triisostearic acid,
octyldodecyl neopentanoic acid, isostearyl octanoic acid, octyl
isononanoic acid, hexyldecyl neodecanoic acid, octyldodecyl
neodecanoic acid, isocetyl isostearic acid, isostearyl isostearic
acid, octyldecyl isostearic acid, polyglycerin oleanoic acid ester,
polyglycerin isostearic acid ester, triisocetyl citric acid,
triisoalkyl citric acid, triisooctyl citric acid, lauryl lactic
acid, myristyl lactic acid, cetyl lactic acid, octyldecyl lactic
acid, triethyl citric acid, acetyltriethyl citric acid, acetyl
tributyl citric acid, trioctyl citric acid, diisostearyl maleic
acid, di 2-ethylhexyl hydroxy stearic acid, 2-ethyl hexyl succinic
acid, diisobutyl adipic acid, diisopropyl sebasinic acid, dioctyl
sebacinic acid, cholesteryl stearic acid, cholesteryl isostearic
acid, cholesteryl hydroxy stearic acid, cholesteryl hydroxy stearic
acid, cholesteryl oleic acid, dihydrocholesteryl oleic acid,
pitsteryl isostearic acid, pitsteryl oleic acid, isocetyl
12-stealoyl hydroxy stearic acid, stearyl 12-stealoyl hydroxy
stearic acid, isostearyl 12-stealoyl hydroxy stearic acid.
93. The method of claim 91, wherein the disease or condition is
selected from the group consisting of: atopic dermatitis, asthma,
food allergy, allergic rhinitis, and chronic urticaria.
94. The method of claim 91, wherein the hardy kiwifruit preparation
is selected from the group consisting of: a fruit extract or
concentrate, a leaf extract or concentrate, a stem extract or
concentrate, or concentrate, a root extract or concentrate, fresh
fruit, crushed fruit, boiled fruit, cooked fruit, pressed fruit,
condensed fruit, dried fruit, a hardy kiwifruit juice concentrate,
a preparation produced by extraction of fruit in water having a
temperature of from 0.degree. C. to about 80.degree. C.; a
preparation produced by direct extraction of a water soluble
concentrate of hardy kiwifruit with ethyl acetate, a preparation
produced by extraction of hardy kiwifruit in distilled water, and a
preparation produced by sequential extraction of hardy kiwifruit in
water, chloroform and ethyl acetate.
95. A method to treat a disease or condition that is associated
with dysregulation of immune function, comprising administering to
a mammal a combination of a hardy kiwifruit or a preparation
thereof and an agent selected from the group consisting of: fatty
acids; citric acid; fumaric acid; steroids; linear peptides;
carbohydrates; herbal preparations; spices; minerals; sterilizers;
seasonings; vitamins; and electrolytes.
96. The method of claim 95, wherein the fatty acids and fatty acid
esters are selected from the group consisting of: glyceryl
tri-2-ethyl hexanoic acid, cetyl 2-ethyl hexanoic acid, isopropyl
myristic acid, butyl myristic acid, isopropyl palmitic acid, ethyl
stearic acid, octyl palmitic acid, isocetyl isostearic acid, butyl
stearic acid, ethyl linoleic acid, isopropyl linoleic acid, ethyl
oleic acid, isocetyl myristic acid, isostearyl myristic acid,
isostearyl palmitic acid, octyldodecyl myristic acid, isocetyl
isostearic acid, diethyl sebasic acid, isopropyl adipic acid,
isoalkyl neopetanoic acid, glyceryl tri(capryl, capric acid),
trimethylopropane tri-2-ethyl hexanoic acid, trimethylopropane
triisostearic acid, pentaerythritol tetra-2 ethyl hexanoic acid,
cetyl caprylic acid, decyl lauric acid, hexyl lauric acid, decyl
myristic acid, myristyl myristic acid, cetyl myristic acid, stearyl
stearic acid, decyl oleic acid, cetyl licinoleic acid, isostearyl
lauric acid, isotridecyl myristic acid, isocetyl palmitic acid,
octyl stearic acid, isocetyl stearic acid, isodecyl oleic acid,
octyldodecyl oleic acid, octyldodecyl linoleic acid, isopropyl
isostearic acid, cetostearyl 2-ethyl hexanoic acid, stearyl 2-ethyl
hexanoic acid, hexyl isostearic acid, ethylene glycol dioctanoic
acid, ethylene glycol dioleic acid, propylene glycol dicapric acid,
propylene glycol di(capryl, capric acid), propylene glycol
dicaprylic acid, neopentylglycol dicapric acid, neopentylglycol
dioctanoic acid, glyceryl tricaprylic acid, glyceryl triundecylic
acid, glyceryl triisopalmitic acid, glyceryl triisostearic acid,
octyldodecyl neopentanoic acid, isostearyl octanoic acid, octyl
isononanoic acid, hexyldecyl neodecanoic acid, octyldodecyl
neodecanoic acid, isocetyl isostearic acid, isostearyl isostearic
acid, octyldecyl isostearic acid, polyglycerin oleanoic acid ester,
polyglycerin isostearic acid ester, triisocetyl citric acid,
triisoalkyl citric acid, triisooctyl citric acid, lauryl lactic
acid, myristyl lactic acid, cetyl lactic acid, octyldecyl lactic
acid, triethyl citric acid, acetyltriethyl citric acid, acetyl
tributyl citric acid, trioctyl citric acid, diisostearyl maleic
acid, di 2-ethylhexyl hydroxy stearic acid, 2-ethyl hexyl succinic
acid, diisobutyl adipic acid, diisopropyl sebasinic acid, dioctyl
sebacinic acid, cholesteryl stearic acid, cholesteryl isostearic
acid, cholesteryl hydroxy stearic acid, cholesteryl hydroxy stearic
acid, cholesteryl oleic acid, dihydrocholesteryl oleic acid,
pitsteryl isostearic acid, pitsteryl oleic acid, isocetyl
12-stealoyl hydroxy stearic acid, stearyl 12-stealoyl hydroxy
stearic acid, isostearyl 12-stealoyl hydroxy stearic acid.
97. The method of claim 95, wherein the disease or condition is
selected from the group consisting of: atopic dermatitis, asthma,
food allergy, allergic rhinitis, and chronic urticaria.
98. The method of claim 95, wherein the hardy kiwifruit preparation
is selected from the group consisting of: a fruit extract or
concentrate, a leaf extract or concentrate, a stem extract or
concentrate, a root extract or concentrate, fresh fruit, crushed
fruit, boiled fruit, cooked fruit, pressed fruit, condensed fruit,
dried fruit, a hardy kiwifruit juice concentrate, a preparation
produced by extraction of fruit in water having a temperature of
from 0.degree. C. to about 80.degree. C.; a preparation produced by
direct extraction of a water soluble concentrate of hardy kiwifruit
with ethyl acetate, a preparation produced by extraction of hardy
kiwifruit in distilled water, and a preparation produced by
sequential extraction in water, chloroform and ethyl acetate.
99. A method to treat a disease or condition that is associated
with dysregulation of immune function, comprising administering to
a mammal a combination of hardy kiwifruit or a preparation thereof
and an agent selected from the group consisting of: taurine; fatty
acids and fatty acid esters.
100. The method of claim 99, wherein the disease or condition is
selected from the group consisting of: atopic dermatitis, asthma,
food allergy, allergic rhinitis, and chronic urticaria.
101. The method of claim 99, wherein the hardy kiwifruit
preparation is selected from the group consisting of: a fruit
extract or concentrate, a leaf extract or concentrate, a stem
extract or concentrate, a root extract or concentrate, fresh fruit,
crushed fruit, boiled fruit, cooked fruit, pressed fruit, condensed
fruit, dried fruit, a hardy kiwifruit juice concentrate, a
preparation produced by extraction of fruit in water having a
temperature of from 0.degree. C. to about 80.degree. C.; a
preparation produced by direct extraction of a water soluble
concentrate of hardy kiwifruit with ethyl acetate, a preparation
produced by extraction of hardy kiwifruit in distilled water, and a
preparation produced by sequential extraction in water, chloroform
and ethyl acetate.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation patent application of
U.S. Provisional Application No. 60/405,295 filed on Aug. 23, 2002,
which was now abandoned.
DESCRIPTION
[0002] 1. Field of the Invention
[0003] The present invention relates to an extract of Actinidia
arguta and related species and a composition comprising the same
having preventing and treating activity of allergic disease and
non-allergic inflammatory disease.
[0004] 2. Background of the Invention
[0005] Allergic diseases such as anaphylaxis, allergic rhinitis,
asthma, atopic dermatitis, food allergies and urticaria, inflict up
to 20% of the population in many countries and are increasing in
prevalence (Wuthrich B., Int. Arch. Allergy Appl. Immunol., 90, pp
3-10, 1989).
[0006] Allergic diseases are mediated by immunoglobulin E (IgE),
while the type-2 T helper (Th2) cell, mast cell and eosinophil are
proven to play important roles in that process (Maggi E.,
Immunotechnology, 3, pp 233-244, 1998; Pawankar R., Curr. Opin.
Allergy Clin. Immunol., 1, pp 3-6, 2001; Vercelli D., Clin. Allergy
Immunol., 16, pp 179-196, 2002).
[0007] Without helminth infection or other stimuli, IgE is normally
one of abundant immunoglobulin isotypes found in human serum as
well as several species of experimental animals (Maggi E.,
Immunotechnology, 3, pp 233-244, 1998; Coffman R L and Carty J., J.
Immunotechnology, 136, pp 949-954, 1986). According to the "Th2
hypothesis", IgE production is favored in the immunological
condition in which humoral immunity mediated by Th2 cells and
related cytokines such as IL-4, IL-5 and IL-13, is predominant
(Maggi E., Immunotechnology, 3, pp 233-244, 1998). The current view
of the progress of allergic diseases is that genetic and
environmental factors interact with each other, leading to the
production of IL-4 through the Stat6-mediated signaling pathway and
the activation of specific transcription factors such as c-Maf,
GATA3, NP45 and NFATc in native T cells, and eventually resulting
in the development of allergen-specific T helper 2 CD4+ cells. Once
generated, allergen-activated Th2 cells secrete IL-4, IL-5 and
IL-13. IL-4 and IL-5 induce the production of IgE and IgG1 by B
cells as well as the stimulation of the development of eosinophils
in bone marrow and their recruitment into inflamed tissues (Erb K.
J.; Immunol. Today, 20, pp 317-322, 1999; Rothenberg M E., N. Engl.
J. Med., 338, pp 1592-1600, 1998). IL-13 is a cytokine closely
related to IL-4 and binds to the IL4 receptor alpha chain inducing
allergic phenotypes independently of IL-4, IgE or eosinophils
(Wills-Karp M., et al., Science, 282, pp 2258-2261, 1998; Grunig
G., et al., Science, 282, pp 2261-2263, 1998).
[0008] Circulating IgE binds to two isoforms of IgE receptors:
high-affinity IgE receptors (FC.epsilon.RI) present on the surface
of mast cells and basophils, and low affinity IgE receptors
(Fc.epsilon.838 RII or CD23) present on the surfaces of
lymphocytes, eosinophils, platelets and macrophages. It is believed
that the most important factor governing the pathogenesis of
allergic disorders is the cross-linkage of IgE receptors on mast
cells, after encountering allergen and the consequent degranulation
of mast cells. The molecules released by mast cells include
histamine, heparin, proteases and free radicals, which mediate a
variety of biological effects including vasodilation, intestinal
and/or bronchial smooth muscle contraction, mucous secretion and
local proteolysis. Following initial immediate reaction of the mast
cells, an influx of eosinophils, basophils and lymphocytes occurs
6-24 hours later. This late-phase response can lead to chronic
tissue inflammation continuously exposed to antigens.
[0009] Conventional drugs for the treatment of allergic disorders
include anti-histamines, steroidal or non-steroidal
anti-inflammatory drugs and leukotriene antagonists. These drugs
are useful mainly for symptomatic effects, and fail to provide with
such treatments that the fundamental cure of allergic diseases such
as alleviating excessive humoral immunity or suppressing IgE
production is required. The hypothesis that reducing serum IgE
level could improve allergic symptoms was demonstrated by clinical
trials of the chimeric anti-IgE antibody (CGP-51901) and
recombinant humanized monoclonal antibody (rhuMAB-E25) (Fahy J V et
al., Am. J. Respir. Crit. Care. Med., 155, pp 1828-1834, 1997).
Diacyl benzimidazole analogs and bacterial polysaccharides that
inhibit IgE synthesis and secretion have been described in U.S.
Pat. No. 6,369,091 and U.S. Patent Application No. 20020041885,
respectively.
[0010] The only method for findamental treatment of allergy is
carrying out immunotherapy or desensitization therapy. The
inununotherapy is treatment method, which reduces hypersensitivity
to allergic origin and improves allergic symptom by administering
refined allergen for long period to allergic patient with gradually
increasing their dosage. After their introduction in 1911, the
immunotherapy has been used for treatment of allergic disease such
as allergic rhinitis, allergic asthma, bee poisoning and so on by
using antigen-specific IgE antibody. Major mechanism reducing
hypersensitivity has not been clearly found yet, but it is known
that increase of IgG concentration while reducing IgE concentration
induces normal immunity reaction.
[0011] Actinidia arguta, A. polygama, and A. kolomikta belonged to
Actinidiaceae, are distributed in Siberia, the northern area of
China, North and South Korea. More than 30 species belonged to
Actinidiaceae has been reported. Among those, the fruit of A.
chinensis or A. delicious have been named as kiwi and Actinidia
arguta and other same genus fruit have been used as materials of
Chinese medicine named as `mihudo` to treat liver disease,
gastrointestinal disease and urogenital lithiasis without toxicity
(Seoul National University Natural Products Science, Tradi-Medi
Data Base, dongbang media Co. Ltd. 1999). However, there have been
no report or suggestion about the treatment and prevention of
allergic disease and non-allergic inflammatory disease using by
Actinidia fruit.
[0012] Meanwhile, there has been concentrated effort to investigate
effective anti-allergy and anti-inflammatory natural products.
[0013] Korea Patent Application No. 92-11752 disclosed an
anti-inflammatory, anti-allergic and anti-rheumatic drug comprising
biflavonoid such as 4'-O-methyl ochnaflavone isolated from Lonicera
japonica, which shows various allergy or inflammation treating
activity. Korea Patent Registration No. 100744 disclosed
anti-inflammatory, anti-allergic and anti-rheumatic drug comprising
several biflavonoid compounds isolated from the leaves of Ginko
biloba. Several Oriental medicine recipes comprising Siegesbeckia
glabrescens have been reported to have IgE-reducing activity (Kim
H. M et al., Phytother: Res., 15, pp 572-576, 2001). Furthermore,
lots of medicinal herbs have been found to be rich sources of
histamine release inhibitors or anti-inflammatory drugs.
[0014] However, there has been not reported or disclosed about
anti-allergic and anti-inflammatory action of hardy kiwifruit
extract in any of above literatures.
[0015] To investigate an anti-allergic and anti-inflammatory drugs
among Chinese herbs, the inventors of the present invention have
intensively carried out in vivo and in vitro experiments concerning
the effects of hardy kiwifruit extract on the change of Th1/T2
cytokines and IgE, IgG subtype in human serum as well as inhibition
test of histamine release from mice peritoneal mast cells and rat
paw edema test.
SUMMARY OF THE INVENTION
[0016] The present invention provides a pharmaceutical composition
comprising the extract of hardy kiwifruit as an active ingredient
in an effective amount to treat and prevent allergic disease and
non-allergic inflammatory disease by reducing inflammation action,
by inhibiting histamine release from mast cell, and by increasing
the level of Th1 cytokines, IgG2a in serum and reducing the level
of Th2 cytokines and IgE in serum.
[0017] The present invention also provides a use of above extract
for the preparation of pharmaceutical composition to treat and
prevent allergic disease and non-allergic inflammatory disease in
mammal or human.
[0018] The present invention also provides a health food or food
additives comprising above extract for prevention or alleviation of
allergic disease and non-allergic inflammatory disease by reducing
inflammation action, by inhibiting histamine release from mast
cell, and by increasing the level of Th1 cytokines, IgG2a in serum,
and reducing the level of Th2 cytokines and IgE in serum.
[0019] The present invention also provides a feed or feed additives
comprising above extract for treatment and prevention of allergic
disease and non-allergic inflammatory disease by reducing
inflammation action, by inhibiting histamine release from mast
cell, and by increasing the level of Th1 cytokines, IgG2a in serum,
and reducing the level of Th2 cytokines and IgE in serum.
[0020] The present invention also provides a cosmetic composition
comprising above extract as an active ingredient in an effective
amount to treat, prevent and improve allergic skin disease and
non-allergic inflammatory skin inflammation disease by reducing
inflammation action, by inhibiting histamine release from mast
cell, and by increasing the level of Th1 cytokines, IgG2a in serum,
and reducing the level of Th2 cytokines and IgE in serum.
BRIEF DESCRIPTION OF THE DRAWINGS
[0021] The above and other objects, features and other advantages
of the present invention will more clearly understood from the
following detailed description taken in conjunction with the
accompanying drawings, in which;
[0022] FIG. 1a shows TLC photograph of the extracts and fractions
of hardy kiwifruit;
[0023] FIG. 1b shows 2D-TLC photograph of [1] sub-fraction;
[0024] FIG. 1c shows 2D-TLC photograph of [2] sub-fraction;
[0025] FIG. 2a presents result about symptom of skin itching, which
investigated at the 12 weeks after administration the hardy
kiwifruit extract in the Nc/Nga mouse having atopic dermatitis;
[0026] FIG. 2b presents result about symptom of skin itching, which
investigated at the 14 weeks after administration the hardy
kiwifruit extract in the Nc/Nga mouse having atopic dermatitis;
DETAILED DESCRIPTION OF THE INVENTION
[0027] Accordingly, it is an object of the present invention to
provide a pharmaceutical composition comprising the crude extract
or non-polar solvent soluble extract of the hardy kiwifruit as an
active ingredients for treatment and prevention of allergic disease
and non-allergic inflammatory disease.
[0028] It is an object of the present invention to provide a use of
a crude extract or non-polar solvent soluble extract of the hardy
kiwifruit for the preparation of therapeutic agent for treatment
and prevention of allergic disease and non-allergic inflammatory
disease in human or mammal.
[0029] It is an object of the present invention to provide a method
of treating or preventing allergic disease and non-allergic
inflammatory disease in a mammal comprising administering to said
mammal an effective amount of crude extract or non-polar solvent
soluble extract of the hardy kiwifruit, together with a
pharmaceutically acceptable carrier thereof.
[0030] It is another object of the present invention to provide a
health food or food additives comprising above extract, together
with a sitologically acceptable additive for prevention and
improvement of allergic disease and non-allergic inflammatory
disease.
[0031] It is still another object of the present invention to
provide an animal feed or feed additives comprising above extract
as essential components for treatment, prevention, and improvement
of allergic disease and non-allergic skin inflammatory disease.
[0032] It is still another object of the present invention to
provide a cosmetic composition comprising above extract for
prevention and improvement of allergic disease and non-allergic
inflammatory disease.
[0033] Above described allergic disease or allergic skin disease
comprise anaphylaxis, allergic rhinitis, asthma, allergic
conjunctivitis, allergic dermnatitis, atopic dermatitis, contagious
dermatitis, urticaria, insect allergies, food allergies and drug
allergies.
[0034] Above described non-allergic skin inflammation disease
comprise various skin trouble caused by inflammation such as
pimple, acne and the like.
[0035] Above described cosmetic composition comprising the hardy
kiwifruit extract having preventing and improving activity of skin
inflammation.
[0036] Above described pharmaceutical composition for treatment and
prevention of allergic disease can be used for the purpose of
increasing the efficiency of allergy immunotheraphy.
[0037] Accordingly, the present invention also provides a
pharmaceutical composition comprising the crude extract or
non-polar solvent soluble extract of the hardy kiwifruit as active
ingredients for allergic immunotheraphy helper.
[0038] Also, the present invention also provides a pharmaceutical
composition comprising an effective amount of the crude extract or
non-polar solvent soluble extract of hardy kiwifruit for treatment
and prevention of non-allergic inflammatory disease.
[0039] Above described non-allergic inflammatory disease comprise
various dermatitis, Systemic Lupus Erythematosus (SLE), retinal
inflammation, gastritis, retinopathy, hepatitis, enteritis,
pancreatitis, nephritis and so on.
[0040] Above hardy kiwifruit may comprises Actinidia arguta, A.
kolomikta, A. polygama or and the same genus plant and may use the
fruit, stem and root thereof
[0041] Above crude extract of hardy kiwifruit can be obtained by
using water, lower alcohols such as methanol, ethanol and the like,
or the mixtures thereof, preferably distilled water or 70% ethanol
soluble extract and above non-polar solvent soluble extract
therefrom can be obtained by using non polar solvent such as
hexane, ethyl acetate or dichloromethane solvent.
[0042] The pharmaceutical composition of the present invention can
contain about 0.01.about.50% by weight of the above extract based
on the total weight of the composition.
[0043] The health food of the present invention comprises above
extracts as 0.01 to 80%, preferably 1 to 50% by weight based on the
total weight of the composition.
[0044] Above health food can be contained in health food, health
beverage etc, and may be used as powder, granule, tablet, chewing
tablet, capsule, beverage etc.
[0045] An inventive extract from the hardy kiwifruit may be
prepared in accordance with the following preferred embodiment.
[0046] Hereinafter, the present invention is described in
detail.
[0047] An inventive extract of hardy kiwifruit can be prepared in
detail by following procedures,
[0048] The inventive crude extract of hardy kiwifruit can be
prepared by follows; hardy kiwifruit is dried and crushed; crushed
hardy kiwifruit is mixed with 5 to 25-fold, preferably,
approximately 10 fold volume of distilled water, lower alcohols
such as methanol, ethanol, butanol and the like, or the mixtures
thereof, preferably water or 70% ethanol; the solution is treated
with hot water at the temperature ranging from 20 to 100.degree.
C., preferably from 60 to 100.degree. C., for the period ranging
from 1 to 24 hours with extraction method by the extraction with
hot water, cold water, reflux extraction, or ultra-sonication, with
1 to 5 times, preferably 2 to 3 times, consecutively; the residue
is filtered to obtain the supernatant to be concentrated with
rotary evaporator, at the temperature ranging from 20 to
100.degree. C., preferably from 50 to 70.degree. C. and then dried
by vacuum freeze-drying, hot air-drying or spray drying to obtain
dried crude extract powder of the hardy kiwifruit which can be
soluble in water, lower alcohols, or the mixtures thereof.
[0049] The above crude extract of the hardy kiwifruit is stored in
-20.degree. C. to use as a sample by dissolving in distilled water
to adjust to certain concentration.
[0050] Additionally, non-polar solvent soluble extract of present
invention can be prepared by following procedure; the crude extract
prepared by above step, is suspended with water, and then is mixed
with 1 to 100-fold, preferably, 1 to 5-fold volume of non polar
solvent such as ethyl acetate, chloroform, hexane and the like; the
non-polar solvent soluble layer is collected to obtain non-polar
solvent soluble extract of the present invention.
[0051] Also, above described procedures may be modified or
subjected further step to fractionate or isolate to obtain more
effective fractions or compounds by the procedure well-known in the
art, for example, the procedure disclosed in the literature
(Harborne J. B., A guide to modern techniques of plant analysis,
3.sup.rd Ed. pp 6-7, 1998).
[0052] To investigate the anti-allergic and anti-inflammatory
activity of the hardy kiwifruit extract prepared by above
procedure, in vivo and in vitro experiments such as ELISA method to
determine the level of Th1/Th2 cytokines and IgE, IgG subtype in
serum, inhibition test of histamine release from mast cells and
anti-inflammation assay to test the effects of inventive extract
was carried out and then it is confirmed that inventive extract
shows excellent anti-allergic and anti-inflammatory effect.
[0053] Specifically, the reduction of allergen specific-IgE and the
increase of allergen specific-IgG2a has been main purpose in
immunotherapy field, the only one present fundamental treatment of
allergic disease and through above experiments, therefore, it is
confirmed that hardy kiwifruit can increase treating efficiency if
it is used with immunotherapy as an allergic immunotheraphy
aid.
[0054] In accordance with another aspect of the present invention,
there is provided a pharmaceutical composition comprising the hardy
kiwifrit extract prepared by above preparation method for the
treatment and prevention of allergic disease and non-allergic
inflammation disease as active ingredients.
[0055] It is another of the present invention to provide a treating
and preventing method comprising administering a pharmaceutical
composition comprising said extract prepared by above preparation
method to allergic disease and non-allergic inflammatory disease of
mammals including human.
[0056] The composition for treating and preventing allergic disease
and non-allergic inflammatory disease may comprises above extracts
as 0.01.about.50% by weight based on the total weight of the
composition.
[0057] The inventive composition may additionally comprise
conventional carrier, adjuvant or diluents in accordance with
conventional using method well known in the art.
[0058] Hereinafter, the following formulation methods and
excipients are merely exemplary and in no way limit the
invention.
[0059] The composition according to the present invention can be
provided as a pharmaceutical composition containing
pharmaceutically acceptable carriers, adjuvants or diluents, e.g.,
lactose, dextrose, sucrose, sorbitol, mannitol, xylitol,
erythritol, maltitol, starches, acacia rubber, alginate, gelatin,
calcium phosphate, calcium silicate, cellulose, methyl cellulose,
polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy
benzoate, talc, magnesium stearate and mineral oil. The
formulations may additionally include fillers, anti-agglutinating
agents, lubricating agents, wetting agents, flavoring agents,
emulsifiers, preservatives and the like. The compositions of the
invention may be formulated so as to provide quick, sustained or
delayed release of the active ingredient after their administration
to a patient by employing any of the procedures well known in the
art.
[0060] For example, the compositions of the present invention can
be dissolved in oils, propylene glycol or other solvents that are
commonly used to produce an injection. Suitable examples of the
carriers include physiological saline, polyethylene glycol,
ethanol, vegetable oils, isopropyl myristate, etc., but are not
limited to them. For topical administration, the compounds of the
present invention can be formulated in the form of ointments and
creamns.
[0061] Pharmaceutical formulations containing present composition
may be prepared in any form, such as oral dosage form (powder,
tablet, capsule, soft capsule, aqueous medicine, syrup, elixirs
pill, powder, sachet, granule), or topical preparation (cream,
ointment, lotion, gel, balm, patch, paste, spray solution, aerosol
and the like), or injectable preparation (solution, suspension,
emulsion).
[0062] The composition of the present invention in pharmaceutical
dosage forms may be used in the form of their pharmaceutically
acceptable salts, and also may be used alone or in appropriate
association, as well as in combination with other pharmaceutically
active compounds.
[0063] The desirable dose of the inventive extract or composition
varies depending on the condition and the weight of the subject,
severity, drug form, route and period of administration, and may be
chosen by those skilled in the art. However, in order to obtain
desirable effects, it is generally recommended to administer at the
amount ranging 10 g/kg, preferably, 1 to 3 g/kg by weight/day of
the inventive extract of the present invention. The dose may be
administered in single or divided into several times per day. In
terms of composition, the amount of inventive extract should be
present between 0.01 to 95% by weight, preferably 0.5 to 80% by
weight based on the total weight of the composition.
[0064] The pharmaceutical composition of present invention can be
administered to a subject animal such as mammals (rat, mouse,
domestic animals or human) via various routes. All modes of
administration are contemplated, for example, administration can be
made orally, rectally or by intravenous, intramuscular,
subcutaneous, intracutaneous, intrathecal, epidural or
intracerebroventricular injection.
[0065] Also, the present invention provide a composition of the
health food beverage for prevention and improvement of allergic
disease and non-allergic inflammation disease adding the hardy
kiwifruit 0.01 to 20% by weight, amino acids 0.001 to 5% by weight,
vitamins 0.001 to 2% by weight, sugars 0.001 to 20% by weight,
organic acids 0.001 to 10% by weight, sweetener and flavors of
proper amount.
[0066] Above the extract of hardy kiwi fruit can be added to food
and beverage for the prevention and improvement of allergic disease
and non-allergic inflammatory disease.
[0067] To develop for health food, examples of addable food
comprising above extracts of the present invention are e.g.,
various food, beverage, gum, vitamin complex, health improving food
and the like, and can be used as power, granule, tablet, chewing
tablet, capsule or beverage etc.
[0068] Also, the extract of present invention will be able to
prevent, and improve allergic disease and non-allergic inflammation
disease by comprising to child and infant food, such as modified
milk powder, modified milk powder for growth period, modified food
for growth period.
[0069] Above described composition therein can be added to food,
additive or beverage, wherein, the amount of above described
extract in food or beverage may generally range from about 0.1 to
95 w/w %, preferably 1 to 80 w/w % of total weight of food for the
health food composition and 1 to 30 g, preferably 3 to 10 g on the
ratio of 100 ml of the health beverage composition.
[0070] Providing that the health beverage composition of present
invention contains above described extract as an essential
component in the indicated ratio, there is no particular limitation
on the other liquid component, wherein the other component can be
various deodorant or natural carbohydrate etc such as conventional
beverage. Examples of aforementioned natural carbohydrate are
monosaccharide such as glucose, fructose etc; disaccharide such as
maltose, sucrose etc; conventional sugar such as dextrin,
cyclodextrin; and sugar alcohol such as xylitol, and erydiritol
etc. As the other deodorant than aforementioned ones, natural
deodorant such as taumatin, stevia extract such as levaudioside A,
glycyrrhizin et al., and synthetic deodorant such as saccharin,
aspartam et al., may be useful favorably. The amount of above
described natural carbohydrate is generally ranges from about 1 to
20 g, preferably 5 to 12 g in the ratio of 100 ml of present
beverage composition.
[0071] The other components than aforementioned composition are
various nutrients, a vitamin, a mineral or an electrolyte,
synthetic flavoring agent, a coloring agent and improving agent in
case of cheese chocolate et al., pectic acid and the salt thereof,
alginic acid and the salt thereof, organic acid, protective
colloidal adhesive, pH controlling agent, stabilizer, a
preservative, glycerin, alcohol, carbonizing agent used in
carbonate beverage et al. The other component than aforementioned
ones may be fruit juice for preparing natural fruit juice, fruit
juice beverage and vegetable beverage, wherein the component can be
used independently or in combination. The ratio of the components
is not so important but is generally range from about 0 to 20 w/w %
per 100 w/w % present composition. Examples of addable food
comprising aforementioned extract therein are various food,
beverage, gum, vitamin complex, health improving food and the
like.
[0072] The inventive composition may additionally comprise one or
more than one of organic acid, such as citric acid, fumaric acid,
adipic acid, lactic acid, malic acid; phosphate, such as phosphate,
sodium phosphate, potassium phosphate, acid pyrophosphate,
polyphosphate; natural anti-oxidants, such as polyphenol, catechin,
.alpha.-tocopherol, rosemary extract, vitamin C, green tea extract,
licorice root extract, chitosan, tannic acid, phytic acid etc.
[0073] The above extract of the hardy kiwifruit may be 20 to 90%
high concentrated liquid, power, or granule.
[0074] Similarly, the above extract of the hardy kiwifruit can
comprise additionally one or more than one of lactose, casein,
dextrose, glucose, sucrose and sorbitol.
[0075] Also, in the present invention, there is also provided a
using method of the food additives such as sterilizer, spice,
seasoning, various nutrients, vitamin, a mineral or an electrolyte,
synthetic flavoring agent, a coloring agent and improving agent in
case of cheese chocolate et al., pectic acid and the salt thereof,
alginic acid and the salt thereof, organic acid, protective
colloidal adhesive, pH controlling agent, stabilizer, a
preservative, glycerin, alcohol, carbonizing agent used in
carbonate beverage et al, or as essential component of food
materials.
[0076] Wherein the food additives can be added to food by
deposition, spray, or mixing the ratio of the additives is not so
important but it generally range fiom about 0.01 to 20 w/w % per
100 w/w % present composition. Examples of addable food comprising
aforementioned extract therein are.
[0077] Wherein the food additives can be added to one or one over
food such as fruits, vegetables, food dehydrated foods or cutting
products such as fruits, vegetables; fruit juice, vegetable juices
or the mixture juices thereof; drinks containing acid-beverage;
confectionaries such as cookie, candy, caramel, gum; breads; ice
creams, teas, fermented milk such as yogurt; dairy product, spices,
alcoholic beverages, cans, in-bottles, noodles, processed livestock
products, processed marine products, fermented food, beans food,
cereals food, processed meats, licorices or hubs.
[0078] In accordance with another aspect of the present invention,
there are provided a feed or feed additive essentially comprising
said extract prepared by above preparation method for prevention
and improvement allergic disease and non-allergic disease.
[0079] Above food additives is characterize of mixing amount of 5
to 100 g per 1 kg by weight based on the total dried weight of the
feed.
[0080] Furthermore, the present invention provides a feed
composition comprising above feed additives.
[0081] Also, the present invention also provides a cosmetic
composition comprising an effective amount of the crude extract or
non-polar solvent soluble extract of hardy kiwifruit for prevention
and improvement of allergic disease or non-allergic inflammatory
disease.
[0082] The present cosmetic composition provide cosmetic
composition comprising the above extracts with 0.01 to 30%, more
preferably, 0.01to 5% by the weight of the inventive composition
based on the total weight of the composition for the treatment,
prevention, and improvement allergic skin disease and non-allergic
skin inflammation disease.
[0083] The other components may be a mixture of the ingredients of
a conventional cosmetic composition well known in the art.
[0084] Cosmetic formulations containing above composition may be
prepared in any form such as skin, lotion, cream, essence, toner,
emulsion, pack, soup, shampoo, rinse, cleanser, body washing
solution, washing solution, treatment, gel, balm, spray solution
and the like.
[0085] The cosmetic composition of the present invention can
comprises additional additives selected from the group consisting
of water soluble vitamin, lipid soluble vitamin, peptide polymer,
polysaccharide polymer, sphingolipid and sea-weed extract.
[0086] Preferable water soluble vitamins are any one which can be
mixed with cosmetic, however, various vitamin such as vitamin B1,
B2, B6, pyridoxine, pyridoxine HCl, vitamin B12, pantothenic acid,
nicotinic acid, nicotinamide, folic acid, vitamin C, vitamin H etc,
their salt thereof such as thiamin HCl salt, ascorbic acid Na salt
etc or their derivatives thereof such as ascorbic acid-2-phosphonic
acid Na salt, ascorbic acid-2-phosphonic acid Mg salt are
preferable and those can be obtained by conventional method such as
microbial conversion method, purification method from the microbial
cultivates, enzymatic method or chemical synthetic method.
[0087] Preferable lipid soluble vitamins are any one which can be
mixed with cosmetic, however, various vitamin such as vitamin A,
D2, D3, E (dl-.alpha.-tocopherol, d-.alpha.-tocopherol, d-67
-tocopherol) and their derivatives such as palmitic acid ascorbate,
stearic acid ascorbate, dipalmnitic acid ascorbate, acetic
acid-dl-.alpha.-tocopherol, nicotinic acid dl-.alpha.-tocopherol
vitamin E, dl-pantothenyl alcohol, D-pantothenyl alcohol,
pantothenyl ethylether etc. containing the lipid soluble vitamin
used in examples of present invention are preferable and those can
be obtained by conventional method such as microbial conversion
method, purification method from the microbial cultivates,
enzymatic method or chemical synthetic method
[0088] Preferable peptide polymers are any one which can be mixed
with cosmetic, however, collagen, hydrolysable collagen, gelatin,
elastin, hydrolysable gelatin, keratin etc. containing the peptide
polymer used in examples of present invention are preferable.
[0089] Preferable polysaccharide polymers are any one which can be
mixed with cosmetic, however, hydroxy ethyl cellulose, xanthin gum,
hyaluronic acid Na, chondroitin sulfate or their salt (Na salt etc)
and the like are preferable. For example, chondroitin sulfate or
the salt thereof etc can be used by being purified from mammal or
fishes ordinarily.
[0090] Preferable sphingolipid are any one, which can be mixed with
cosmetic, however, ceramide, pit-sphingosin,
sphingo-lipopolysaccharide and the like are preferable.
Sphingo-lipid can be obtained by being purified from mammal, fish,
shellfish, yeast or plant etc in conventional method.
[0091] Preferable seaweed extract is any one which can be mixed
with cosmetic, however, the extract of brown algae, red algae,
green algae and the like or the purified carrageenan, alginic acid,
arginic acid Na, K isolated therefrom are preferable. Algae extract
can be obtained by being purified from seaweed in conventional
method.
[0092] The cosmetic composition of the present invention may
combine with other ingredients combined with conventional cosmetic
composition, if necessary, together with above described essential
ingredient.
[0093] Preferable above described other ingredients may comprises
oil ingredient, humectants, emollients, surface active agents,
organic or inorganic dye, organic powder, ultraviolet ray absorbing
agent, preservatives, antiseptics, antioxidants, plant extract, pH
controller, alcohol, pigments, perfumes, refrigerants, blood
circulator, antihidrotic, distilled water etc.
[0094] Preferable oil ingredients may comprise ester oil,
hydrocarbon oil, silicone oil, fluoride oil, animal oil, plant oil
and so on.
[0095] Preferable ester oil described above may comprise glyceryl
tri-2-ethyl hexanoic acid, cetyl 2-ethyl hexanoic acid, isopropyl
myristic acid, butyl myristic acid, isopropyl palmitic acid, ethyl
stearic acid, octyl palmitic acid, isocetyl isostearic acid, butyl
stearic acid, ethyl linoleic acid, isopropyl linoleic acid, ethyl
oleic acid, isocetyl myristic acid, isostearyl myristic acid,
isostearyl palmitic acid, octyldodecyl myristic acid, isocetyl
isostearic acid, diethyl sebasic acid, isopropyl adipic acid,
isoalkyl neopetanoic acid, glyceryl tri(capryl, capric acid),
trimethylopropane tri-2-ethyl hexanoic acid, trimethylopropane
triisostearic acid, pentaerythritol tetra-2 ethyl hexanoic acid,
cetyl caprylic acid, decyl lauric acid, hexyl lauric acid, decyl
myristic acid, myristyl myristic acid, cetyl myristic acid, stearyl
stearic acid, decyl oleic acid, cetyl licinoleic acid, isostearyl
lauric acid, isotridecyl myristic acid, isocetyl palmitic acid,
octyl stearic acid, isocetyl stearic acid, isodecyl oleic acid,
octyldodecyl oleic acid, octyldodecyl linoleic acid, isopropyl
isostearic acid, cetostearyl 2-ethyl hexanoic acid, stearyl 2-ethyl
hexanoic acid, hexyl isostearic acid, ethylene glycol dioctanoic
acid, ethylene glycol dioleic acid, propylene glycol dicapric acid,
propylene glycol di(capryl, capric acid), propylene glycol
dicaprylic acid, neopentylglycol dicapric acid, neopentylglycol
dioctanoic acid, glyceryl tricaprylic acid, glyceryl triundecylic
acid, glyceryl triisopalmitic acid, glyceryl triisostearic acid,
octyldodecyl neopentanoic acid, isostearyl octanoic acid, octyl
isononanoic acid, hexyldecyl neodecanoic acid, octyldodecyl
neodecanoic acid, isocetyl isostearic acid, isostearyl isostearic
acid, octyldecyl isostearic acid, polyglycerin oleanoic acid ester,
polyglycerin isostearic acid ester, triisocetyl citric acid,
triisoalkyl citric acid, triisooctyl citric acid, lauryl lactic
acid, myristyl lactic acid, cetyl lactic acid, octyldecyl lactic
acid, triethyl citric acid, acetyltriethyl citric acid, acetyl
tributyl citric acid, trioctyl citric acid, diisostearyl maleic
acid, di 2-ethylhexyl hydroxy stearic acid, 2-ethyl hexyl succinic
acid, diisobutyl adipic acid, diisopropyl sebasinic acid, dioctyl
sebacinic acid, cholesteryl stearic acid, cholesteryl isostearic
acid, cholesteryl hydroxy stearic acid, cholesteryl hydroxy stearic
acid, cholesteryl oleic acid, dihydrocholesteryl oleic acid,
pitsteryl isostearic acid, pitsteryl oleic acid, isocetyl
12-stealoyl hydroxy stearic acid, stearyl 12-stealoyl hydroxy
stearic acid, isostearyl 12-stealoyl hydroxy stearic acid.
[0096] Preferable hydrocarbon oil described above may comprise
squalene, liquid paraffin, .alpha.-olefin oligomer, isoparaffin,
ceresin, paraffin, liquid isoparaffin, polybuden, microcrystalline
wax, vaselin and the like.
[0097] Preferable silicone oil may comprise polymethylsilicone,
methylphenylsilicone, metbylcyclopolysiloxane,
octamethylpolysiloxane, decamethylpolysiloxane,
dodecamnethylcyclosiloxane, dimethyl siloxane-methyl
cetyloxysiloxane copolymer, dimethyl siloxane-methyl
stealloxysiloxane copolymer, alkyl modified silicone oil, amino
modified silicone oil and the like.
[0098] Preferable fluoride oil can comprise perfluoropolyether and
the like.
[0099] Preferable animal or plant oil can comprise avocado oil,
almond oil, olive oil, sesame oil, rice husk oil, safflower oil,
soy-bean oil, corn oil, rape oil, amygdalin oil, palm kernel oil,
palm oil, pimaja oil, sunflower oil, fruite seed oil, cotton seed
oil, coconut palm oil cucui nut oil, wheat embryo bud oil, rice
embryo bud oil, sia butter, evening-primrose oil, marker daymia nut
oil, medo home oil, egg yolk oil, lanolin, hempseed oil, mink oil,
orange ruppy oil, hohoba oil, carnawa wax, liquid lanolin, solid
pimaja wax and the like.
[0100] Preferable humectants can comprise water-soluble low
molecular humectants, lipophilic low molecular humectants,
water-soluble polymer and lipid soluble polymer.
[0101] Specifically, preferable water soluble low molecular
humectants can comprise cerin, glutamine, sorbitol, mannitol,
pyrrolidone-carboxylic acid Na, glycerin, propylene glycol,
1,3-butylene glycol, ethylene glycol, polyethylene glycol
(polymerization index. >2), polypropylene glycol (polymerization
index >2), lactic acid, lactate salt and the like.
[0102] Preferable lipid soluble low molecular humectants can
comprise cholesterol, cholesteryl ester and the like.
[0103] Preferable water-soluble polymer can comprise carboxy vinyl
polymer, poly asparaginic acid salt, tragacanth, xanthin gum, HMC
(hydroxy methyl celluose), HEC (hydroxy ethyl celluose), HPC
(hydroxy propyl celluose), carboxymethylcellulose, water-soluble
chitin, chitosan, dextrin and the like.
[0104] Preferable lipid soluble polymer can comprise
polyvinylpyrrolidone-eicocene copolymer,
polyvinylpyrrolidone-hexadecene copolymer, nitrocellulose, dextrin
fatty acid ester, silicone polymer and the like.
[0105] Preferable emollients can comprise long chain acyl glutamic
acid cholesteryl ester, cholesteryl hydroxy stearic acid,
12-hydroxy stearic acid, rogic acid, lanolin fatty acid cholesteryl
ester and the like.
[0106] Preferable surface-active agent can comprise nonionic
surfactants, anionic surfactants, cationic surfactants, amphivalent
surfactants and the like.
[0107] Specifically, preferable non-ionic surfactants can comprise
self-emulsified monostearic acid glycerin, propylene glycol fatty
acid ester, glycerin fatty acid ester, polyglycerin fatty acid
ester, sorbitan fatty acid ester, polyoxyethylene (POE) sorbitan
fatty acid ester, POE sorbitan fatty acid ester, POE glycerin fatty
acid ester, POE alkyl ether, POE fatty acid ester, POE solid pimaja
oil, POE pimaja oil, POE-POP copolymer, POE-POP alkyl ether,
polyether modified silicone, lauric acid alkanol amide, alkyl amine
oxide, hydrogen addition soybean phospholipid and the like.
[0108] Preferable anionic surfactants can comprise fatty acid soap,
.alpha.-acyl sulfonic acid salt, alkyl sulfonic acid salt, alkyl
ally sulfonic acid, alkyl naphthalene sulfonic acid salt, alkyl
sulfonic acid salt, POE alkylether sulfate salt, alkyl amide
sulfate salt, alkyl phosphate salt, POE alkyl phosphate salt,
alkylamide phospahate salt, alkyloylalkyl taurine salt,
N-acyl-amino acid salt, POE alkyl ether carboxylic acid salt, alkyl
sulfo succinic aid salt, alkyl sulfo-acetic acid salt, acylated
hydrolysable collagen peptide salt, perfluoro alkyl phosphate ester
and the like.
[0109] Preferable cationic surfactant can comprise alkyl trimethyl
ammonium chloride, stearyl trimethyl ammonium chloride, stearyl
trimethyl ammonium bromide, setostearyltrimethyl ammonium chloride,
distearyl dimethyl ammonium chloride, stearyl dimethyl benzyl
ammonium chloride, vehenyltrimethyl ammonium bromide, benzalkonium
chloride, diethylamino ethyl arnide stearic acid,
dimethylaminopropyl amide stearic acid, lanolin derivatives
quatemary ammonium and the like.
[0110] Preferable ambivalent surfactants can comprise carboxy
betaine type, amide betaine type, hydroxy sulfo betaine type,
phosphpobetaine type, aminocarboxylic acid, imidazoline derivatives
type, amide amine type and the like.
[0111] Preferable organic and inorganic dyes can comprise silicic
acid, anhydrous silicic acid, magnesium silicic acid, talc,
ceracyte, mica, kaolin, bengala, clay, bentonite, titan film mica,
oxy chlorine bismuth, zirconium oxide, magnesiun oxide, zinc oxide,
titan oxide, aluminium oxide, calcium sulfate, barium sulfate,
magnesium sulfate, calcium carbonate, magnesium carbonate, ferrous
oxide, chromium oxide, chromium hydroxide, calamine, carbon black
and the combination thereof as an inorganic dyes; polyamide,
polyester, polypropylene, polystyrene, polyurethane, vinyl resin,
urea resin, phenol resin, fluoride resin, silicone resin, acryl
resin, melamine resin, epoxy resin, polycarbonate resin, divinyl
benzene-styrene copolymer, silk powder, cellulose, CI pigmnent
yellow, CI pigment orange as an organic dyes; and their complex
etc.
[0112] Preferable organic powder can comprise metal soap such as
calcium stearate; alkyl phosphonate metal salt such as sodium zinc
cetylic acid, zinc laurylic acid, calcium laurylic acid; acylamino
acid polyvalent metal salt such as calcium
N-lauroyl-.beta.-alanine, zinc N-lauroyl-.beta.-alanine, calcium
N-lauroyl-glycine etc.; amide sulfonic acid polyvalent metal salt
such as calcium N.epsilon.lauroyl-taurine, calcium
N.epsilon.-palmitoyl-taurine; palmitoyl ornitine, N.alpha.-lauroly
arginine, hardened lanolin fatty acid acyl arginine and the like;
N-acylpolypeptide such as N-lauroylglycyl glycine; .alpha.-amino
fatty acid such as .alpha.-amino caprylic acid, .alpha.-amino
lauric acid and the like; polyethylene, polypropylene, nylon,
polymethylmetacrylate, polystyrene, divinylbenzene-styrene
copolymer, ethylene tetrafluoride and so on.
[0113] Preferable ultraviolet absorbing agents can comprise
paraaminobenzoic acid, paraamonoethyl benzoate, paraamino amyl
benzoate, paraamnino octyl benzoate, ethyleneglycol salicylate,
phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl
salicylate, homomentyl salicylate, benzyl cinnamic acid,
paramethoxy 2-ethoxy ethyl cinnamic acid, paramethoxy octyl
cinnamic acid, diparamethoxy mono-2-ethylhexane glyceryl cinnamic
acid, paramethoxy isopropyl cinnamic acid, diisopropyl -diisopropyl
cinnamnate ester mixture, urokanic acid, ethyl urokanic acid,
hydroxy methoxy benzophenone, hydroxymethoxy benzophenone sulfonic
acid and the salt thereof, dihydroxy methoxy benzophenone,
dihydroxy methoxy benzophenone disulfonate Na, dihydroxy
benzophenone, tetrahydroxybenzophenone,
4-tert-butyl-4'-methoxydibenzoylmethane,
2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1,3,5-triazine,
2-(2-hydroxy-5-methylphenyl)benzotriazole and the like.
[0114] Preferable preservatives can comprise hinokitiol, trichloric
acid, trichlorohydroxydiphenylether, chlorohexidine glucuronate,
phenoxyethanol, resorcine, isopropylmethylphenol, azulene,
salicylic acid, zinc pilithione, bezalconium HCl, photosensitizer
301, mononitroguaiacol Na, undecylenic acid etc.
[0115] Preferable antioxidants can comprise butylhydroxyanisole,
propyl gallate, ellisorbate and the like.
[0116] Preferable pH controller can comprise citric acid, sodium
citrate, malic acid, sodium malate, flmaric acid, sodium fumaric
acid, succinic acid, sodium succinic acid, sodium hydroxide, sodium
hydrogen phosphate and the like.
[0117] Preferable alcohol can comprise cetyl alcohol etc.
[0118] Furthermore, other ingredient addable to above described
component and the amount thereof is not limited within the scope of
the purpose and effect of the present invention, however, it is
preferable that the amount of the other ingredients ranges from
0.01 to 5%, more preferably, 0.01 to 3% in that of total
composition.
[0119] The cosmetic composition of the present invention can be
modified as a solution, emulsion, cohesive mixture etc.
[0120] Above described ingredients such as water-soluble vitamin,
lipid soluble vitamin, peptide polymer, polysaccharide polymer,
sphingolipid, sea weed extract and addable ingredients which can be
added other than above described ingredients if necessary, can be
obtained by conventional methods disclosed in the literature
(Matsumoto Mithio, Manual for the development of transdermal
applied preparation. Seisi Press, 1.sup.st Ed., 1985).
[0121] Additionally, the present invention also provides a cosmetic
additives comprising above extract as an essential component for
prevention or improvement of allergic disease and non-allergic
disease.
[0122] Above cosmetic additives can be used by adding to existing
cosmetics and washing solution to prevent, improve or treat
allergic disease and non-allergic skin disease.
[0123] Furthermore, above cosmetic additives can be used to cream,
lotion, message pack, and body washing solution, soup, shampoo and
the like.
[0124] Inventive extract of the present invention have no toxicity
and adverse effect therefor; they can be used with safe.
[0125] It will be apparent to those skilled in the art that various
modifications and variations can be made in the compositions, use
and preparations of the present invention without departing from
the spirit or scope of the invention.
[0126] The present invention is more specifically explained by the
following examples. However, it should be understood that the
present invention is not limited to these examples in any
manner.
EXAMPLES
[0127] The following Examples and Experimental Examples are
intended to further illustrate the present invention without
limiting its scope.
Example 1
Preparation of Hardy Kiwifruit Extract
1-1. Preparation of Water Extract of Hardy Kiwifruit
[0128] 100 g of dried hardy kiwifruit and dried stem of hardy
kiwifruit (Actinidia arguta), dried fruit of A. kolomikta and A.
polygama purchased from Kyung-dong Market located in Seoul was
crushed, mixed with 1 L of distilled water and subjected to reflux
extraction for 3 hrs at 90.about.95.degree. C. with three times and
the extract was filtered with filter paper, concentrated using by
rotary evaporator (N-1000, Eyela Co. Japan) at 55.about.65.degree.
C. under reduced pressure and dried with freezing dryer to obtain
15.6 g of dried fruit extract, 10.4 g of dried stem extract of
kiwifruit (Actinidia arguta), 16.2 g and 17.0 g of dried fruit
extract of A. kolomikta, and A. polygama respectively. The dried
powder was dissolved in distilled water (100 mg/ml).
1-2. Preparation of Water-alcohol Soluble Extract of Hardy
Kiwifruit
[0129] Except using various mix ratio of water-alcohol solvent
mixture such as 30%, 50%, and 70% ethanol solvent with as an
extracting solvent, all the procedure was identical to those of
Example 1-1. As a result, 11 g.about.13 g of dried power of hardy
kiwifruit was obtained at each ratio of solvent mixture and the
dried powder was dissolved in distilled water (100 mg/ml).
Example 2
Preparation of Polar Solvent and Non-polar Solvent Soluble Hardy
Kiwifruit Extract
[0130] The water extract prepared in Example 1-1 was subject to
fractionation by following procedure.
2-1. Preparation of Chloroform Soluble Fraction
[0131] 50 ml of distilled water was added to 5 g of hardy kiwifruit
extract obtained in Example 1-1. 50 ml of chloroform was added
thereto in separatory funnel, shaken vigorously to divide into
chloroform soluble layer and water soluble layer.
2-2. Preparation of Ethyl Acetate Soluble Fraction
[0132] Above water soluble layer obtained in Example 1-1 was mixed
with 50 ml of ethyl acetate and then divided into ethyl acetate
soluble layer and water soluble layer.
[0133] Above chloroform soluble layer, ethyl acetate soluble layer
and water layer were concentrated by rotary evaporator, dried with
freeze dryer to obtain 0.34 g of chloroform soluble fraction, 0.05
g of ethyl acetate soluble fraction and 4.61 g of water fraction
powders respectively.
Example 3
Fractionation of Hardy Kiwifruit Extract by Silica Gel Column
Chromatography
[0134] 2,784 mg of ethyl acetate soluble fraction in Example 2-2
was further subjected to silica gel column chromatography (Daiso
gel IR-60-W-40:63 mm). The developing solvent was started with
chloroform:methanol:water ([1] 90:11:1, [2] 60:10:1, [3] 60:20:2)
solvent mixture and ended with methanol[4] with eluting speed of
300 ml/hr to obtain four sub-fractions([1] 2,381 mg, [2] 135 mg,
[3] 148 mg, [4] 98 mg).
[0135] Above water extract, ethyl acetate soluble fraction and four
sub-fractions were subjected to TLC (TLC plate: Merck Co. Ltd.,
Developing solvent; chloroform:methanol:water=9:5:1) and the
results were shown in FIG. 1a. As shown in FIG. 1a, lane 1 is water
extract, lane 2 is ethyl acetate soluble fraction, lane 3 is [4]
sub-fraction, lane 4 is [3] sub-fraction, lane 5 is [2]
sub-fraction and lane 6 is [1] sub-fraction.
[0136] Above [1] and [2] sub-fractions were subjected to 2D-TLC
using chloroform:methanol:water (9:5:1) solvent mixture as a
1.sup.st developer and chloroform:acetone:water (3:8:0.5) solvent
mixture as a 2.sup.nd developer (See FIG. 1b and FIG. 1c).
Experimental Example 1
Inhibition of IgE Production U266B1 Cell Line by Hardy Kiwifruit
Extract
1-1. Effect of Hardy Kiwifruit Extract on IgE Production
[0137] To confirm the inhibitory effect of hardy kiwifruit
extraction on IgE production, U266B1 cell (lymphoblastoma cell
line) was used. U266B1 is cell line of human B cell, which produces
IgE.
[0138] U266B1 cells (American Type Culture Collection, Manassas,
Va.) was cultured at 37.degree. C. under 5% CO.sub.2 circumstances
in 24-well culture plate containing RPMI-1640 medium (15% FBS, 2mM
L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 50 .mu.g
streptomycin and 100 U/ml penicillin). Cells were adjusted at the
concentration of 2.times.10.sup.5 cell/well and treated with LPS (2
.mu.g/ml) and 100 .mu.g/m of hardy kiwifruit extract prepared in
Example 1-1 and 1-2. Control was treated with 2 .mu.M dexamethasone
or medium (RPMI-1640). After treatment, cells were cultured for 7
days and IgE concentration of medium was measured by ELISA kit
(PharMingen; San Diego, Calif.).
[0139] As shown in the Table 1, all of the water extracts of hardy
kiwifruit, A. kolomikta, and A. polygama showed the inhibitory
effect on IgE production as much as dexamethasone(control). Also,
hardy kiwifriit 70% ethanol extract showed the strongest activity
among all the three kinds of inventive hardy kiwifruit alcohol
extract. The stem extract of hardy kiwifruit showed similar effect
to those of above hardy kiwifruit extracts. TABLE-US-00001 TABLE 1
LPS stimulation Treatment IgE (IU/ml) - Medium 187.8 .+-. 8.4 +
Medium 392.9 .+-. 3.4 + Hardy kiwifruit water extract 228.8 .+-.
7.1 + Actinidia kolomikta water extract 233.2 .+-. 5.8 + Actinidia
polygama water extract 211.7 .+-. 7.9 + Hardy kiwifruit stem
extract 241.5 .+-. 5.8 + Hardy kiwifruit 30% ethanol extract 306.2
.+-. 16.5 + Hardy kiwifruit 50% ethanol extract 266.8 .+-. 17.0 +
Hardy kiwifruit 70% ethanol extract 201.9 .+-. 27.6 + Dexamethasone
203.6 .+-. 30.9
1-2. Effect of Ethyl Acetate Soluble Fraction of Hardy Kiwifruit on
IgE Production
[0140] To compare the inhibitory effect on IgE production of U266B1
cell, chloroform soluble fraction, ethyl acetate soluble fraction
and water soluble fraction which were prepared in above Example 2
were subjected to the identical experiment disclosed in above
Experimental Example 1-1.
[0141] The chloroform soluble fraction, ethyl acetate soluble
fraction and water soluble fraction were treated to U266B1 cell
line (at 30 .mu.g/ml).
[0142] As shown in the Table 2, ethyl acetate soluble fraction
showed stronger inhibitory effect on IgE production than that of
Dexamethasone (control). TABLE-US-00002 TABLE 2 LPS stimulation
Treatment IgE (IU/ml) - Medium 117.4 .+-. 7.6 + Medium 212.5 .+-.
11.8 + Hardy kiwifruit water extract 151.7 .+-. 11.9 + Chloroform
fraction 206.7 .+-. 15.6 + Ethyl acetate fraction 123.3 .+-. 8.1 +
Water fraction 218.9 .+-. 13.3 + Dexamethasone 138.9 .+-. 2.1
1-3. Effect of Silica Gel Column Chromatography Fraction on IgE
Production
[0143] Silica gel column chromatography fraction [1], [2], [3]) and
[4] prepared in Example 3 were subjected to the identical
experiment disclosed in Experimental Example 1-1 to compare the
inhibitory effect of IgE production on U266B1cell.
[0144] Silica gel column chromatography fraction [1], [2], [3] and
[4] were treated to U266B 1 cell line (at 10 .mu.g/ml).
[0145] As the result of Table 3, silica gel column chromatography
fraction [1], [2] showed the inhibitory activity on IgE production.
TABLE-US-00003 TABLE 3 LPS stimulation Treatment IgE (IU/ml) -
Medium 379.7 .+-. 13.2 + Medium 540.4 .+-. 35.1 + Ethyl acetate
fraction 298.1 .+-. 9.7 + Silica gel fraction 1 293.5 .+-. 12.5 +
Silica gel fraction 2 307.6 .+-. 24.1 + Silica gel fraction 3 453.1
.+-. 17.3 + Silica gel fraction 4 396.9 .+-. 26.8 + Dexamethasone
277.6 .+-. 12.4
Experimental Example 2
Anti-allergic Effect of Hardy Kiwifruit Extract in
Ovalbumin-sensitized Mouse Model
2-1. Preparation of Ovalbumin-sensitized Mouse Model
[0146] The ovalbumin-sensitized mouse model is commonly used as
animal model of allergy. Female BALB/c mice, aged 6 weeks (Seoul
national university animal experimental center) were adapted to the
environment for 7 days. At the time of 7 weeks after birth, 100
.mu.l of emulsion, mixed 25 .mu.g of ovalbumin (chicken egg
albumin, crude grade V, Sigma Co., Ltd.) with 2.25 mg of aluminum
hydroxide (ImujectAlum, Pierce Co., Ltd.) was injected into mouse
peritoneal cavity and 14 days after, it was injected once more for
boosting.
[0147] And then 30 mice were divided into 3 groups and each group
was orally administered with the hardy kiwifruit water extract of
Example 1-1(300 .mu.g/mouse/day), dexamethasone (10
.mu.g/mouse/day), or drinking water (100 .mu.g/mouse/day) for 11
days, respectively. On the 25th day, mice were sacrificed and blood
serun and spleen was collected therefrom.
2-2. Concentration Analysis of Ovalbumin-specific IgE, IgG1, IgG2a,
IgG2b in Serum
[0148] In serum collected in Example 2-1, the serum levels of
ovalbumin-specific IgE, IgG1, IgG2a and IgG2b were measured by
ELISA kits (PharMingen Co., Ltd).
[0149] As shown in Table 4, it shows that relative concentration of
each antibody of the ovalbumin-sensitized mouse to that of normal
mouse. In the mouse administered with the water extract of hardy
kiwifluit, ovalbumin-specific IgE level was decreased under 1/3 and
IgG1 level was significantly decreased. The IgE and IgG1 level was
similarly decreased in dexamethasone-administered mouse.
[0150] The IgG2a level associated with cellular immunity, was
increased above 2 times in the mouse administered with the hardy
kiwifruit water extract, however it was not increased in the
dexamethasone-administered mouse.
[0151] It showed that the hardy kiwifruit extract could improve
fundamental allergic constitution by decreasing IgE, which induces
allergic symptoms, and increasing IgG2a related to normal immunity
simultaneously. It is also clear that hardy kiwifluit can increase
treatment efficiency when it is used with immunotherapy as an
allergic immunotheraphy helper since the reduction of allergen
specific-IgE and the increase of allergen specific-IgG2a have been
their main purpose in immunotherapy field. TABLE-US-00004 TABLE 4
Drinking Hardy kiwifruit Normal water extract Dexamethasone IgE 1.0
.+-. 0.0 7.6 .+-. 0.2 2.2 .+-. 0.5 2.0 .+-. 0.6 IgG1 1.0 .+-. 0.0
5.1 .+-. 0.1 3.3 .+-. 0.3 3.1 .+-. 0.3 IgG2a 1.0 .+-. 0.1 2.1 .+-.
0.4 4.6 .+-. 0.3 1.9 .+-. 0.4 IgG2b 1.0 .+-. 0.0 1.4 .+-. 0.1 1.5
.+-. 0.2 1.2 .+-. 0.1 unit: pg/ml
2-3. Expression Analysis of Cytokines by Splenocytes
[0152] To analyze the expression of cytokines by the splenocytes of
mouse administered with the hardy kiwifruit extract, splenocytes
was prepared from spleen prepared in Example 2-1 as follows.
[0153] Each prepared splenocytes was pooled and homogenized under
an aseptic condition.
[0154] Splenocytes were washed with RPMI-1640 medium, filtered
through nylon mesh (60 .mu.m pore size) to eliminate large clots,
centrifuged (1500 rpm, 5 mins) to separate a precipitated cells,
and then cells were added to RPMI-1640 medium supplemented with 10%
FBS.
[0155] Splenocytes prepared as above were inoculated in 24-well
plate (5.times.10.sup.6 cell/ml/well), treated with 100 .mu.g/ml of
ovalbumin, and incubated at 37.degree. C. under an atmosphere
containing 5% C0.sub.2for 3 days. After cultivation was finished, a
culture solution was gathered and then the concentration of
cytokines (IL-4, IL-5, IL-12 and interferon-.gamma.) related to
allergy were measured by commercial ELISA kits.
[0156] As shown in the Table 5, splenocytes of mouse administered
with the hardy kiwifruit extract had the decreased level of IL and
IL-5 (which comes to Th2 cytokines that is to induce allergy) and
has the increased level of IL -12 and interferon-.gamma. (which
comes to Th1 cytokines that is to repress allergy). In the case of
dexamethasone-treated mouse, both of Th1 and Th2 cytokines were
decreased. TABLE-US-00005 TABLE 5 OVA-stimulated mouse Normal
Drinking Hardy kiwifruit mouse water extract Dexamethasone IL-4
21.6 .+-. 11.8 159.5 .+-. 15.7 76.5 .+-. 10.0 23.2 .+-. 8.7 IL-5
0.5 .+-. 0.8 2573.6 .+-. 42.0 1638.3 .+-. 33.3 884.1 .+-. 80.7
IL-12 1626.0 .+-. 58.5 906.2 .+-. 66.0 1321.2 .+-. 92.4 297.7 .+-.
16.4 Interferon-.gamma. 14.1 .+-. 19.6 1016.0 .+-. 25.6 1688.2 .+-.
15.8 470.5 .+-. 38.6 unit: pg/ml
[0157] It is confirmed that the hardy kiwifit extract can prevent
and improve allergic disease by increasing Th1 cytokines as well as
decreasing Th2 cytokines specifically, while dexamethasone
suppresses the whole immune system.
Experimental Example 3
Down Regulation of Th1/Th2 Cytokines in Human PBMC
[0158] Human peripheral blood mononuclear cells (PBMCs) were
prepared with Ficoll-hypaque from the whole blood of an allergy
patient having a high basal serum level of IgE and cultured in RPMI
media with 10% FBS. PBMCs were treated together with the hardy
kiwifruit extract (at 100 .mu.g/ml) and phytohemagglutinin (PHA, at
5 .mu.g/ml), a commonly used lectin with an immune stimulating
effect, and cultured at 5% CO.sub.2 and 37.degree. C. Forty-eight
hours later, the level of IL-5, IL-13, IL10 and Interferon-.gamma.
in cell culture supernatant was measured using ELISA.
[0159] Table 6 shows that the hardy kiwifruit extract significantly
reduced the serum level of Th2 cytokines: the levels of IL-5 and
IL-13 were reduced by 52% and 47%, respectively, whereas the serum
level of Interferon-.gamma., a Th1 cytokine, was increased by 3.2
folds. This result suggested that hardy kiwifruit extract could
increase the level of Th1 cytokines, while simultaneously
decreasing the level of Th2 cytokines. It has been previously
reported that down-regulation of Th2 cytokines could contribute to
the alleviation of IgE synthesis and allergic inflammation.
TABLE-US-00006 TABLE 6 PHA stimulation Treatment IL-5 IL-13 IL-10
Interferon-.gamma. - -- 92.2 .+-. 6.4 0 .+-. 0 0 .+-. 0 53.3 .+-.
30.2 + Medium 518.1 .+-. 120.3 667.8 .+-. 46.5 480.3 .+-. 15.3
334.1 .+-. 277.7 + Hardy 248.5 .+-. 62.0 355.7 .+-. 93.1 570.2 .+-.
56.3 1067 .+-. 345.1 kiwifruit extract unit: pg/ml
Expedmental Example 4
Reduction of the Human Serum Level of IgE
[0160] To test whether hardy kiwifruit extract reduces the serum
level of IgE in humans or not, two allergy patients (Patients K and
E with allergic rhinitis and allergic dermatitis, respectively,
showing high basal serum levels of IgE) were orally administered
with hardy kiwifruit extract (1 g in dry weight) on a daily basis
over a 21-day period, and their serum levels of IgE were measured
every two weeks by ELISA method.
[0161] At the result of experiment, it was shown that the serum
levels of IgE in the two patients were reduced continuously by 2/3
after 42 days and the dermatitis symptom of patient E were also
improved during the period of experiment (See Table 7).
TABLE-US-00007 TABLE 7 1 day 14 day 28 day 42 day Patient K 950.8
IU/ml 855.6 IU/ml 679.1 IU/ml 266.1 IU/ml Patient E 278.3 IU/ml
236.0 IU/ml 189.2 IU/ml 95.0 IU/ml
Experimental Example 5
Inhibition of Histamine Release from Mice Peritoneal Mast Cells
[0162] The release of histamine from mast cells is one of the major
causes of allergic reactions. Therefore, the effects of hardy
kiwifruit extract were tested on histamine release from mast
cells.
[0163] Each mouse was anesthetized with ether, and injected with 20
ml of Tyrode buffer B (NaCl, glucose, NaHCO.sub.3, KCl,
NaH.sub.2PO.sub.4) containing 0.1% gelatin into the peritoneal
cavity; then the abdomen was gently massaged for about 90 seconds.
The peritoneal cavity was carefully opened, and the fluid
containing peritoneal cells was aspirated by pasteur pipette. The
peritoneal cells were then sedimented at 150.times.g for 10 min. at
room temperature and resuspended in Tyrode buffer B. Mast cells
were separated from the major components of rat peritoneal cells as
described in the literature (Yurt et al., J Exp Med., 1;146(5), pp
1405-19, 1977). Peritoneal cells suspended in 1 ml of Tyrode buffer
B were layered onto 2 ml of 0.225 g/ml metrizamide (density 1.120
g/ml) and centrifuged at room temperature for 15 min at
400.times.g. The cells remaining at the buffer-metrizamide
interface were aspirated and discarded; the cells in the pellet
were washed and resuspended in 1 ml of Tyrode buffer A containing
calcium. Splenocytes were seeded into 24-well culture plates
(2.times.10.sup.5 cells/well) in 0.4 ml medium for each well. Cells
were incubated overnight at 37.degree. C. and sensitized with 0.5
.mu.g/ml of anti-DNP.sub.24-BSA IgE. After sensitizing the cells
with IgE, the medium was removed, and the cells were washed twice
with 0.5 ml of PIPES buffer and preincubated with either 200 .mu.l
of PIPES buffer (as control), Cromolyn (10.sup.-4 M) or hardy
kiwifruit extract (100 .mu.g/ml) at 37.degree. C. for 10 min. Mast
cells were stimulated with 20 ng/ml of DNP.sub.24-BSA as an antigen
for 30 min., and histamine released into the medium was measured by
ELISA (ALerCHEK). Inhibition of histamine release was calculated as
following empirical formula 1; Percent
inhibition=100.times.(A-B)/(A-C) [Empirical Formula 1] [0164] A:
Stimulated level (Histamine release with IgE-stimulation). [0165]
B: Suppressed level (Histamine release with IgE-stimulation and
drug treatment). [0166] C: Basal level (Histamine release without
IgE-stimulation)
[0167] Table 8 indicates that the hardy kiwifruit extract inhibited
histamine release by approximately 44% at 100 .mu.g/ml, and it was
effective as much as Cromolyn used as a positive control.
TABLE-US-00008 TABLE 8 Treatment Cromolyn Hardy kiwifruit
Inhibition (%) 52 .+-. 13 44 .+-. 10
Experimental Example 6
[0168] Anti-inflammatory Effects on Edema Induced by Arachidonic
Acid in the Ears of Mice
[0169] 15 mice (8 week old male BALB/c) were fasted for 18 hours
with free access to water and divided into 3 groups. Inflammation
was induced by topical application of arachidonic acid (0.5 mg/20
.mu.l acetone) to the right ear of each mouse. The left ear was
used as a negative control and received the vehicle (20 .mu.l
acetone). The hardy kiwifruit extract (200 mg/kg in water) was
administered p.o. 1 hour prior to arachidonic acid application. The
positive control group received indomethacin (10 mg/kg, p.o.) 1
hour before arachidonic acid application. Inflammation was followed
for 1 hour and thereafter the animals were sacrificed. A section of
6 mm diameter disc from each ear was obtained and weighed. The
edema index was assessed using the increase in the weight of the
treated right ear punch biopsy over that of the untreated left ear.
The edema index of the control mice that did not receive any
treatment was 7.2.+-.1.1 mg. However, when animals were treated
with the hardy kiwifruit extract, the index was decreased by 62.5%
to 2.7.+-.0.8 mg. The ear treated with indomethacin as a positive
control showed 81.9% decrease in edema index compared to the
untreated ear (See Table 9).
[0170] This data suggested that the hardy kiwifruits had
anti-inflammatory activity comparable to indomethacin in this
experimental model. TABLE-US-00009 TABLE 9 Treatment Index of edema
Inhibition rate (%) Control 7.2 .+-. 1.1 -- Hardy kiwifruit 2.7
.+-. 0.8 62.5 extract Indomethacin 1.3 .+-. 0.3 81.9
Experimental Example 7
[0171] Experiment of Mouse Model with Allergic Dermatitis
[0172] To confirm anti-allergic effect of hardy kiwifruit extract
in animals, Nc/Nga mouse model that has been widely used as an
animal model for human atopic dermatitis study was employed. Nc/Nga
mouse has suppressed Th1 immunity because of it's genetic
character, i.e., low level of interferon-.gamma. production and
consequently, Th2 immunity becomes dominant, which predisposes
Nc/Nga mouse to allergic disease, notably atopic dermatitis under
normal circumstance (Vestergaar C H et al., J. Clin. Invest., 104,
pp 1097-1105, 1999).
[0173] For experiment, 15 of Nc/Nga mice (7 weeks after birth,
female) were divided into 3 groups and adapted to new circumstance
for 1 week. Since 8 weeks after birth, the hardy kiwifruit extract
(300 .mu.g/mouse/day) prepared in example 1-1 as a treatment group
had been orally administered and dexamethasone (10 .mu.g/mouse/day)
or drinking water (100 .mu.g/mouse/day) as control groups had been
orally administered for 8 weeks. To compare the progress of
dermatitis symptom, the scratching frequency of mouse (the time
spent scratching during 20 min-observation) was measured when mice
is grown at the age of 12 and 14 weeks. 16 weeks old mouse was
sacrificed and the quantity of IgE, IgG1 and IgG2a level in serum
were measured by ELISA method. Also, to compare the production of
Th1/Th2 cytokines, splenocyte was prepared by conventional method
from each mouse, transferred into 24-well plate (5.times.10.sup.6
cells/ml/well) and incubated with ConA (1 .mu.g/ml) for 3 days and
then the levels of IL-4, IL-5, IL-12, and Interferon-.gamma. were
measured by ELISA method.
[0174] It was confirmed that the hardy kiwifruit extract repressed
itching symptom as much as dexamethasone, a steroidal
antiphlogistic agent, when the frequency of scratching was measured
(See FIG. 2a: the result of 12 weeks after birth, FIG. 2b: the
result of 14 weeks after birth).
[0175] Table 10 shows that the serum level of IgE of the hardy
kiwifruit extract-administered mouse was reduced remarkably.
[0176] In the Table 11, the production of IL-4 and IL-5 was
decreased and the production of interferon-.gamma. and IL-12 was
increased obviously in the splenocytes, which were prepared from
the hardy kiwifruit extract-administered mouse. The hardy kiwifruit
extract improved itching symptom, lowered IgE concentration in
serum, and repressed the Th2 cytokine production by splenocytes
(similarly with dexamnethasone), but unlike dexamethasone, the
hardy kiwifruit extract increased significantly IL-12 and
interferon-.gamma. level, which have been well known to contribute
to alleviation of allergic diseases. TABLE-US-00010 TABLE 10 IgE
IgG1 IgG2a Drinking water 1572.9 .+-. 77.4 242.2 .+-. 14.2 177.5
.+-. 17.2 Hardy kiwifruit 699.0 .+-. 348.5 263.4 .+-. 21.1 263.5
.+-. 16.2 extract Dexamethasone 130.0 .+-. 55.3 247.3 .+-. 10.8
148.0 .+-. 14.8 unit: pg/ml
[0177] TABLE-US-00011 TABLE 11 IL-4 IL-5 IL-12 Interferon-.gamma.
Drinking water 151.9 .+-. 2.2 778.3 .+-. 34.1 1925.7 .+-. 134.5
10407.5 .+-. 130.8 Hardy kiwifruit 24.1 .+-. 5.2 248.0 .+-. 17.8
2346.2 .+-. 98.4 15847.9 .+-. 1693.1 extract Dexamethasone 42.7
.+-. 19.4 646.5 .+-. 51.7 201.2 .+-. 12.1 10096.4 .+-. 192.4 unit:
pg/ml
Experimental Example 8
Toxicity Test
[0178] To examine the toxicity of the hardy kiwifruit extract,
repetitive toxicity tests were performed on mouse.
[0179] The 10 female of Balb/c mice were divided into 2 groups and
the inventive hardy kiwifruit extract (150 mg/kg) was administered
to the mice at 150 mg/kg for 4 weeks and water was administered to
the control group. The symptom of toxicity was observed for 4 weeks
such as the change of weight, the hematological analysis and
histological test.
[0180] As a result of experiment, there was no death example of the
mice administered with 150 mg/kg inventive hardy kiwifruit and
there was no significant abnormality in the gain of weight, the
caloric intake of feed, the hematological analysis or the
histological test etc. In accordance with above results, it was
confirmed that the hardy kiwifruit was safe. [0181] (1) Weight and
behavior observation: the unusual change of weight or behavior was
not observed. [0182] (2) Hematological analysis: No abnormal
symptom was observed in the number of WBC, lymphocyte, monocyte,
neutrophil, eosinophil, basophil, RBC, hemoglobin or platelet.
[0183] (3) Serum biochemical test: No abnormal symptom was observed
in the level of AST, ALT, LDH, bilirubin, creatinine, glucose,
cholesterol, minerals, albumin, BUN, lipase or amylase of serum.
[0184] (4) Histological test: No abnormal symptom was observed in
the tissue of kidneys, the spleen, the liver or the thymus.
[0185] Hereinafter, the formulating methods and kinds of excipients
will be described, but the present invention is not limited to
them. The representative preparation examples were described as
follows. TABLE-US-00012 Preparation of injection Hardy kiwifruit
water extract of Example 1 50 mg Sodium metadisulfite 3.0 mg
Methylparaben 0.8 mg Propylparaben 0.1 mg Distilled water for
injection optimum amount
[0186] Injection preparation was prepared by mixing above
components and making 2 ml by the conventional method and then
filing filling all the components 2 ml ample and sterilizing by
conventional injection preparation method. TABLE-US-00013
Preparation of tablet Hardy kiwifruit water extract of Example 1 50
mg Corn Starch 100 mg Lactose 100 mg Magnesium Stearate 2 mg
[0187] Tablet preparation was prepared by mixing above components
and entabletting. TABLE-US-00014 Preparation of capsule Hardy
kiwifruit water extract of Example 1 100 mg Corn starch 100 mg
Lactose 100 mg Talc 2 mg Magnesium Stearate optimum amount
[0188] Tablet preparation was prepared by mixing above components
and filling gelatin capsule by conventional gelatin preparation
method. TABLE-US-00015 Preparation of liquid The hardy kiwifruit
70% ethanol extract 100 mg Sugar 20 g Fructose 20 g Lemon flavour
optimum amount Distilled water 100 ml
[0189] Liquid preparation was prepared by mixing above components
and then filling 100 ml brown bottle sterilizing by conventional
liquid preparation method. TABLE-US-00016 Preparation of health
care food Hardy kiwifruit water extract of Example 1 1000 mg
Vitamin mixture 20 g Vitamin A acetate 70 .mu.g Vitamin E 1.0 mg
Vitamin B.sub.1 0.13 mg Vitamin B.sub.2 0.15 mg Vitamin B6 0.5 mg
Vitamin B12 0.2 .mu.g Vitamin C 10 mg Biotin 10 .mu.g Amide
nicotinic acid 1.7 mg Folic acid 50 .mu.g Calcium pantothenic acid
0.5 mg Mineral mixture optimum amount Ferrous sulfate 1.75 mg Zinc
oxide 0.82 mg Magnesium carbonate 25.3 mg Monopotassium phosphate
15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium
carbonates 100 mg Magnesium chloride 24.8 mg
[0190] The above-mentioned vitamin and mineral mixture may be
varied in many ways. Such variations are not to be regarded as a
departure from the spirit and scope of the present invention.
TABLE-US-00017 Preparation of health beverage Hardy kiwifruit water
extract of Example 1 1000 mg Citric acid 100 mg Oligosaccharide 100
g Apricot concentration 2 g Taurine 1 g Distilled water 900 ml
[0191] Health beverage preparation was prepared by dissolving
active component, mixing, stirred at 85.degree. C. for 1 hour,
filtered and then filling all the components in 2000 ml ample and
sterilizing by conventional health beverage preparation method.
TABLE-US-00018 Preparation of skin lotion Hardy kiwifruit water
extract of Example 1 1.00(%) Glycerol 3.00 Ethanol 1.00 Propylene
glycol 0.10 Flavour trace amount Distilled water made to 100%
[0192] Skin preparation was prepared by dissolving active component
according to conventional lotion preparation method. TABLE-US-00019
Preparation of lotion Hardy kiwifruit water extract of Example 1
3.00(%) L-ascorbic acid-2-magnesium phosphate 1.00 Soluble collagen
(1% solution) 1.00 Sodium citric acid 0.10 Citric acid 0.05
1,3-butylene glycol 3.00 Distilled water made to 100%
[0193] Lotion preparation was prepared by dissolving active
component according to conventional lotion preparation method.
TABLE-US-00020 Preparation of cream Hardy kiwifruit water extract
of Example 1 3.00(%) Polyethyleneglycomonosterate 2.00 Monostearate
glycerin 1.00 Cetyl alcohol 4.00 Squalene 6.00 Tri 2-glyceryl
ethylhexanoate 6.00 Sphingo-glycolipid 1.00 1,3-butylene glycol
7.00 Distilled water made to 100%
[0194] TABLE-US-00021 Preparation of pack Hardy kiwifruit water
extract of Example 1 .sup. 5.00(%) Polyvinyl alcohol 13.00
L-ascorbic acid-2-magnesium phosphate 1.00 Lauroylhydroxyproline
1.00 Soluble collagen (1% solution) 2.00 1,3-butylene glycol 3.00
Ethanol 5.00 Distilled water made to 100%
[0195] Pack preparation was prepared by dissolving active component
according to conventional pack preparation method. TABLE-US-00022
Preparation of beauty solution Hardy kiwifruit water extract of
Example 1 .sup. 2.00(%) Hydroxyethylenecellulose (2% solution)
12.00 Xanthin gum (2% solution) 2.00 1,3-butylene glycol 3.00 Conc.
Glycerin 4.00 Sodium hyaluronate 5.00 Distilled water made to
100%
[0196] Beauty solution preparation was prepared by dissolving
active component according to conventional beauty solution
preparation method.
[0197] The invention being thus described, it will be obvious that
the same may be varied in many ways. Such variations are not to be
regarded as a departure from the spirit and scope of the present
invention, and all such modifications as would be obvious to one
skilled in the art are intended to be included within the scope of
the following claims
INDUSTRIAL APPLICABILITY
[0198] As described in the present invention, an extract of the
hardy kiwifruit extract prepared by inventive preparation increase
serum levels of Th1 cytokines and IgG2a, reduce serum levels of Th2
cytokines and IgE, inhibits histamine release from mast cells, and
suppresses inflammatory reaction. According to this, the hardy
kiwifruit can be used as a pharmaceutical composition for the
treatment and prevention of allergic diseases, such as anaphylaxis,
allergic rhinitis, asthma, atopic dermatitis, food allergies and
urticaria and non-allergic inflammation disease.
[0199] Furthermore, an extract of the hardy kiwifruit extract can
be used as a composition of healthy food for the treatment and
prevention of allergic diseases, such as anaphylaxis, allergic
rhinitis, asthma, atopic dernatitis, food allergies and urticaria
and non-allergic inflammation disease.
* * * * *