U.S. patent application number 14/361243 was filed with the patent office on 2015-03-05 for pharmaceutical composition for prevention or treatment of cognitive function disorders comprising spinosyn.
The applicant listed for this patent is DAE HWA PHARMA. CO. LTD, MINISTRY OF FOOD AND DRUG SAFETY. Invention is credited to Sam Sik Kang, Do Hoon Kim, Dong Sup Kim, Ju Sun Kim, Seung Hee Kim, Hyung Eun Lee, So Young Lee, Jong Hoon Ryu.
Application Number | 20150065441 14/361243 |
Document ID | / |
Family ID | 48535792 |
Filed Date | 2015-03-05 |
United States Patent
Application |
20150065441 |
Kind Code |
A1 |
Ryu; Jong Hoon ; et
al. |
March 5, 2015 |
PHARMACEUTICAL COMPOSITION FOR PREVENTION OR TREATMENT OF COGNITIVE
FUNCTION DISORDERS COMPRISING SPINOSYN
Abstract
The pharmaceutical composition of the present invention, which
comprises spinosin or a spinosin-containing herbal extract, is a
composition for preventing and treating cognitive function
disorders, and can effectively improve memory and learning ability,
and effectively treat and prevent cognitive function disorders such
as dementia and amnesia.
Inventors: |
Ryu; Jong Hoon; (Seoul,
KR) ; Lee; Hyung Eun; (Seoul, KR) ; Kang; Sam
Sik; (Seoul, KR) ; Kim; Ju Sun; (Seongnam-si,
KR) ; Lee; So Young; (Seoul, KR) ; Kim; Do
Hoon; (Seoul, KR) ; Kim; Dong Sup; (Seoul,
KR) ; Kim; Seung Hee; (Seoul, KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MINISTRY OF FOOD AND DRUG SAFETY
DAE HWA PHARMA. CO. LTD |
Cheongwon-gun
Hoengseong-gun |
|
KR
KR |
|
|
Family ID: |
48535792 |
Appl. No.: |
14/361243 |
Filed: |
November 30, 2012 |
PCT Filed: |
November 30, 2012 |
PCT NO: |
PCT/KR2012/010318 |
371 Date: |
October 13, 2014 |
Current U.S.
Class: |
514/27 ;
536/18.1 |
Current CPC
Class: |
A61K 36/48 20130101;
A61K 36/47 20130101; A61P 25/28 20180101; A61K 36/42 20130101; A61K
36/725 20130101; A23L 33/135 20160801; A23L 2/52 20130101; A61K
36/185 20130101; A23L 33/10 20160801; A23V 2002/00 20130101; A61K
31/7048 20130101 |
Class at
Publication: |
514/27 ;
536/18.1 |
International
Class: |
A61K 31/7048 20060101
A61K031/7048; A23L 1/30 20060101 A23L001/30 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 30, 2011 |
KR |
10-2011-0127108 |
Claims
1. A pharmaceutical composition comprising spinosin for prevention
or treatment of cognitive impairments.
2. The pharmaceutical composition of claim 1, wherein the spinosin
is obtained from at least one extract selected from the group
consisting of Zizyphus jujuba Mill var. inermis, Zizyphus jujuba
Mill var. hoonensis, Zizyphus jujuba Mill var. spinosa, Passiflora
edulis flavicarpa, Cayaponia tayuya, Desmodium tortuosum,
Wilbrandia ebracteata, Strophioblachia fimbricalyx, Clutia
abyssinica, and Saccharopolyspora spinosa.
3. The pharmaceutical composition of claim 1, wherein the cognitive
impairments comprise dementia and amnesia.
4. A pharmaceutical composition comprising spinosin for prevention
or treatment of cognitive impairments, wherein the pharmaceutical
composition comprises at least one extract selected from the group
consisting of Zizyphus jujuba Mill var. inermis, Zizyphus jujuba
Mill var. hoonensis, Zizyphus jujuba Mill var. spinosa, Passiflora
edulis flavicarpa, Cayaponia tayuya, Desmodium tortuosum,
Wilbrandia ebracteata, Strophioblachia fimbricalyx, Clutia
abyssinica, and Saccharopolyspora spinosa.
5. The pharmaceutical composition of claim 4, wherein the extract
is obtained from the seeds of Ziziphus jujuba Mill var.
spinosa.
6. A food composition comprising spinosin for prevention or
treatment of cognitive impairments.
7. A food composition comprising spinosin for prevention or
improvement of cognitive impairments, wherein the food composition
comprises at least one extract selected from the group consisting
of Zizyphus jujuba var. inermis, Zizyphus jujuba var. hoonensis,
Zizyphus jujuba var. spinosa, Passiflora edulis flavicarpa,
Cayaponia tayuya, Desmodium tortuosum, Wilbrandia ebracteata,
Strophioblachia fimbricalyx, Clutia abyssinica, and
Saccharopolyspora spinosa.
Description
TECHNICAL FIELD
[0001] The present invention relates to a pharmaceutical
composition comprising spinosin for prevention and treatment of
cognitive impairments. More particularly, the present invention
relates to a pharmaceutical composition which can improve memory
and learning ability, thus effectively preventing and treating
disorders such as dementia and amnesia.
BACKGROUND ART
[0002] With the development of medical technology and the
improvement of living standards, the average lifespan of a human
has been almost doubled over the past half century, and thus the
ratio of the elderly population to the total population has
increased rapidly. Along with the development of an aging society,
senile dementia has emerged as one of the major health problems
that need to be addressed in the twenty-first century. Therefore,
the need to develop functional materials, foods, etc. that can
prevent and treat cognitive impairments including dementia has been
increasing.
[0003] Dementia that is a typical disease of cognitive impairments
is a pathological condition that needs to be distinguished from
normal aging and is classified into Alzheimer's disease, vascular
dementia, and other dementias caused by alcoholism, trauma,
sequelae of Parkinson's disease, etc.
[0004] It has been reported that Alzheimer's disease is one of
chronic psychiatric disorders including disorders of higher
cerebral functions such as loss of memory, impaired consciousness,
spatiotemporal chaos, thinking ability, arithmetic ability,
judgment, common sense, etc. Moreover, it has been reported that
Alzheimer's disease also occurs in relatively young people and most
frequently occurs in elderly people as the incidence is increased
two times for each increase of 5 years in the range of 65 to 85
years old. While the pathogenesis of Alzheimer's disease is not
clearly known, a decrease in acetylcholine function in the central
nervous system most commonly occurs, and thus therapeutic methods
of administering acetylcholine precursors or drugs that inhibit the
degradation of acetylcholine to increase the concentration of
acetylcholine in the brain have been used. Therefore,
acetylcholinesterase (hereinafter, `AChE`) inhibitors have been
used alone or in combination with existing cholinesterase
inhibitors, and examples of such drugs include tacrine, donepezil,
rivastigmine, galantamine, etc. While these drugs are
acetylcholinesterase inhibitors, they only slow the progression of
diseases, but have little effect on the treatment and have limited
therapeutic potential at the beginning of the disease, and thus
made efforts have been made to develop drugs that treat the
underlying cause of Alzheimer's disease (Terry and Buccafusco,
2003; Kar et al., 2004; Akhondzadeh et al., 2008; Cummings et al.,
2008; Voss et al., 2008).
[0005] Vascular dementia is mostly caused by damage to the brain
cells due to lack of blood supply to certain parts of the brain
caused by cerebral arteriosclerosis. The causes of the vascular
dementia and Alzheimer's disease are different, but they are the
same in that they cause damage to memory and learning ability.
[0006] Accordingly, the present inventors have studied to develop
an effective medicine for prevention or treatment of cognitive
impairments such as dementia and found herbal extracts and their
active ingredients which can effectively improve memory and
learning ability, thus completing the present invention.
PRIOR ART LITERATURES
Patent Literatures
[0007] KR 10-2010-0034779; [0008] KR 10-2004-0083414; [0009] KR
10-2003-0062963.
Non-Patent Literatures
[0009] [0010] 2009 Alzheimer's disease facts and figures,
Alzheimer's Dement (2009) 5(3): p. 234-70; [0011] Whitehouse, P.
J., Price, D. L., Struble, R. G., Clark, A. W., Coyle, J. T., and
Delong, M. R. (1982) Alzheimer's disease and senile dementia: loss
of neurons in the basal forebrain. Science 215, 1237-12392; [0012]
Hyman, B. T., Van Hoesen, G. W., Damasio, A. R., and Barnes, C. L.
(1984) Alzheimer's disease: cell-specific pathology isolates the
hippocampal formation. Science 225, 1168-11703.
DISCLOSURE
Technical Problem
[0013] An object of the present invention is to provide a
pharmaceutical composition comprising spinosin for prevention or
treatment of cognitive impairments.
[0014] Another object of the present invention is to provide a
pharmaceutical composition comprising a spinosin-containing herbal
extract for prevention or treatment of cognitive impairments.
[0015] Still another object of the present invention is to provide
a food composition comprising spinosin for prevention or
improvement of cognitive impairments.
[0016] Yet another object of the present invention is to provide a
food composition comprising a spinosin-containing herbal extract
for prevention or improvement of cognitive impairments.
[0017] Still yet object of the present invention is to provide a
pharmaceutical composition comprising spinosin for prevention or
treatment of degenerative brain diseases.
[0018] A further object of the present invention is to provide a
pharmaceutical composition comprising a spinosin-containing herbal
extract for prevention or treatment of degenerative brain
diseases.
[0019] Another further object of the present invention is to
provide a method for preventing or treatment of cognitive
impairments or degenerative brain diseases, the method comprising
administering to a subject in need thereof a composition comprising
spinosin.
Technical Solution
[0020] The present invention relates to a pharmaceutical
composition comprising spinosin for prevention or treatment of
cognitive impairments.
[0021] Spinosin
(6-(2-O-beta-D-Glocopyranosyl-beta-D-glucopyranosyl)-5-hydroxy-2-(4-hydro-
xyphenyl)-7-methoxy-4H-1-benzopyran-4-one) is a compound
represented by the following formula 1:
##STR00001##
[0022] Spinosin represented by the above formula 1 can prevent or
treat cognitive impairments. More particularly, the spinosin
represented by the above formula 1 can improve memory and learning
ability, thus effectively preventing and treating cognitive
impairments.
[0023] The pharmaceutical composition comprising spinosin of the
present invention can significantly improve learning ability, space
perception ability, and memory in animal models with memory
impairment induced by scopolamine, and thus exhibits excellent
activity in the prevention or treatment of cognitive impairments
and is particularly useful for the improvement of dementia and
amnesia.
[0024] Spinosin represented by the above formula 1 may be obtained
from various types of herbal medicines. For example, the spinosin
represented by the above formula 1 may be obtained from Zizyphus
jujuba Mill var. inermis, Zizyphus jujuba Mill var. hoonensis,
Zizyphus jujuba Mill var. spinosa, Passiflora edulis flavicarpa,
Cayaponia tayuya, Desmodium tortuosum, Wilbrandia ebracteata,
Strophioblachia fimbricalyx, Clutia abyssinica, Saccharopolyspora
spinosa, or mixtures thereof, and may preferably be obtained from
zizyphi spinosi semen, the seeds of Zizyphus jujuba Mill var.
spinosa.
[0025] Spinosin represented by the above formula 1 may be obtained
from an extract of the above-described herbal medicines or mixtures
thereof, and the spinosin represented by the above formula 1 may
preferably be obtained from a water, methanol, ethanol, butanol, or
hexane extract of the above-described herbal medicines or mixtures
thereof. For example, the spinosin represented by the above formula
1 may be obtained from an extract of zizyphi spinosi semen, the
seeds of Ziziphus jujuba Mill var. spinosa.
[0026] Preferably, the spinosin represented by the above formula 1
may be obtained from an extract of an ethanol extract of zizyphi
spinosi semen, the seeds of Zizyphus jujuba Mill var. spinosa.
[0027] Otherwise, Spinosin represented by the above formula 1 may
be commercially available.
[0028] According to the present invention, the cognitive
impairments refer to diseases caused by impaired functions such as
memory, space perception ability, judgment, executive function,
linguistic ability, etc. and may include, for example, Alzheimer's
disease, vascular dementia, other dementias caused by various
factors alcoholism, trauma, sequelae of Parkinson's disease, etc.,
or amnesia. Preferably, the cognitive impairment may be Alzheimer's
disease.
[0029] The pharmaceutical composition comprising spinosin of the
present invention can effectively inhibit scopolamine-induced
memory impairments in mice even with a small amount.
[0030] The spinosin is an ingredient contained in the
above-described herbal medicines and can effectively prevent or
treat cognitive impairments without side effects.
[0031] In the pharmaceutical composition comprising spinosin for
prevention or treatment of cognitive impairments according to the
present invention, the spinosin may be administered once or several
times at a daily dose of about 1 mg to 120 mg for an adult,
preferably at a daily dose of 30 mg to 120 mg. However, the dose of
the spinosin can be appropriately adjusted depending on the
condition of a patient, such as the patient's severity, age, sex,
weight, etc., and the formulation, administration route, and
administration period.
[0032] The present invention relates to a pharmaceutical
composition for prevention and treatment of cognitive impairments,
comprising spinosin represented by the above formula 1 and an
extract of Zizyphus jujuba Mill var. inermis, Zizyphus jujuba Mill
var. hoonensis, Zizyphus jujuba Mill var. spinosa, Passiflora
edulis flavicarpa, Cayaponia tayuya, Desmodium tortuosum,
Wilbrandia ebracteata, Strophioblachia fimbricalyx, Clutia
abyssinica, Saccharopolyspora spinosa, or mixtures thereof.
[0033] The above-mentioned Zizyphus jujuba Mill var. inermis,
Zizyphus jujuba Mill var. hoonensis, Zizyphus jujuba Mill var.
spinosa, Passiflora edulis flavicarpa, Cayaponia tayuya, Desmodium
tortuosum, Wilbrandia ebracteata, Strophioblachia fimbricalyx,
Clutia abyssinica, and Saccharopolyspora spinosa comprise spinosin
represented by the above formula 1. Therefore, the composition
comprising an extract of the above-described herbal medicines can
prevent or treat cognitive impairments, like the spinosin, and has
no side effects because it is a herbal extract.
[0034] The pharmaceutical composition of the preset invention may
comprise an extract of Zizyphus jujuba Mill var. spinosa,
preferably a water, hexane, ethanol, methanol, butanol, or the
mixture thereof extract of the seeds of Ziziphus jujuba Mill var.
spinosa, more preferably an ethanol extract.
[0035] The spinosin-containing herbal extract can effectively
prevent or treat cognitive impairments without side effects.
[0036] The herbal extract of the present invention comprises
spinosin and thus can significantly improve learning ability, space
perception ability, and memory in animal models with memory
impairment induced by scopolamine, and thus exhibits excellent
activity in prevention or treatment of cognitive impairments and is
particularly useful for the improvement of dementia and
amnesia.
[0037] The pharmaceutical composition of the present invention may
comprise the herbal extract in an amount of 0.1 to 50 wt % with
respect to the total weight of the composition. However, the
content is not necessarily limited thereto, but may vary depending
on the patient's condition and the type and progression of the
disease.
[0038] In the pharmaceutical composition comprising a herbal
extract for prevention or treatment of cognitive impairments
according to the present invention, the herbal extract may be
administered once or several times at a daily dose of about 10 mg
to 1200 mg for an adult, preferably at a daily dose of 300 mg to
1200 mg. However, the dose of the herbal extract may be
appropriately adjusted depending on the condition of a patient,
such as the patient's severity, age, sex, weight, etc., and the
formulation, administration route, and administration period.
[0039] The pharmaceutical composition comprising the spinosin
represented by the above formula 1 or the spinosin-containing
herbal extract has no toxicity and side effects and thus can be
used with confidence even when taken for a long time for the
purpose of prevention or treatment.
[0040] The present invention relates to a pharmaceutical
composition comprising a pharmaceutically acceptable salt of
spinosin for prevention or treatment of cognitive impairments or
degenerative brain diseases. The term "pharmaceutically acceptable
salt" refers to salts commonly used in the pharmaceutical industry,
such as inorganic ion salts formed with potassium, sodium,
magnesium, etc., inorganic acid salts formed with hydrochloric
acid, nitric acid, phosphoric acid, bromic acid, iodic acid,
perchloric acid, tartaric acid, sulfuric acid, etc., organic acid
salts formed with acetic acid, trifluoroacetic acid, citric acid,
maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric
acid, fumaric acid, mandelic acid, propionic acid, lactic acid,
glycolic acid, gluconic acid, galacturonic acid, glutamic acid,
glutaric acid, glucuronic acid, aspartic acid, ascorbic acid,
carbonic acid, vanillic acid, hydroiodic acid, etc., sulfonic acid
salts formed with methanesulfonic acid, ethanesulfonic acid,
benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic
acid, etc., amino acid salts formed with glycine, arginine, lysine,
etc., and amine salts formed with trimethylamine, triethylamine,
ammonia, pyridine, picoline, etc., but the types of salts used in
the present invention are not limited to the above-mentioned
salts.
[0041] The cognitive impairments may be dementia or amnesia, and
the degenerative brain diseases may be Parkinson's disease, stroke,
stroke, or Huntington's disease.
[0042] The pharmaceutical composition of the present invention may
further comprise pharmaceutically acceptable additives such as
diluents, binders, disintegrants, lubricants, pH-adjusting agents,
antioxidants, and solubilizers within the range where effects of
the present invention are not impaired.
[0043] The diluents may include sugar, starch, microcrystalline
cellulose, lactose (lactose hydrate), glucose, D-mannitol,
alginate, alkaline earth metal salt, clay, polyethylene glycol,
calcium anhydrous hydrogen phosphate, and mixtures thereof; and the
binders may include starch, microcrystalline cellulose, highly
dispersive silica, mannitol, D-mannitol, sucrose, lactose hydrate,
polyethylene glycol, polyvinylpyrrolidone (povidone),
polyvinylpyrrolidone copolymer (copovidone), hypromellose,
hydroxypropylcellulose, natural gum, synthetic gum, copovidone,
gelatin, and mixtures thereof.
[0044] The disintegrants may include starches or modified starches
such as sodium starch glycolate, corn starch, potato starch,
pregelatinized starch, etc.; clays such as bentonite,
montmorillonite, veegum, etc.; celluloses such as microcrystalline
cellulose, hydroxypropylcellulose, carboxymethylcellulose, etc.;
algins such as sodium alginate, alginic acid, etc.; crosslinked
celluloses such as croscarmellose sodium, etc.; gums such as guar
gum, xanthan gum, etc.; crosslinked polymers such as crosslinked
polyvinylpyrrolidone (crospovidone), etc.; effervescent agents such
as sodium bicarbonate, citric acid, etc.; and mixtures thereof.
[0045] The lubricants may include talc, stearic acid, magnesium
stearate, calcium stearate, sodium lauryl sulfate, hydrogenated
vegetable oil, sodium benzoate, sodium stearyl fumarate, glyceryl
behenate, glyceryl monorate, glyceryl monostearate, glyceryl
palmitostearate, colloidal silicon dioxide, and mixtures
thereof.
[0046] The pH-adjusting agents may include acidifying agents such
as acetic acid, adipic acid, ascorbic acid, sodium ascorbate,
sodium etherate, malic acid, succinic acid, tartaric acid, fumaric
acid, and citric acid, and basifying agents such as precipitated
calcium carbonate, aqueous ammonia, meglumine, sodium carbonate,
magnesium oxide, magnesium carbonate, sodium citrate, and tribasic
calcium phosphate.
[0047] The antioxidants may include dibutyl hydroxy toluene,
butylated hydroxyanisole, tocopherol acetate, tocopherol, propyl
gallate, sodium hydrogen sulfite, and sodium pyrosulfite. The
solubilizers that may be used in a prior-release compartment of the
present invention may include sodium lauryl sulfate,
polyoxyethylene sorbitan fatty acid esters such as polysorbate,
docusate sodium, poloxamer, etc.
[0048] For oral administration, the pharmaceutical composition
comprising spinosin or the spinosin-containing herbal extract
according to the present invention may be formulated into solid
dosage forms such as tablets, pills, powders, granules, capsules,
etc., and these solid dosage forms may be prepared by mixing
spinosin or the spinosin-containing herbal extract with one or more
excipients such as starch, calcium carbonate, sucrose or lactose,
gelatin, etc. Moreover, lubricants such as magnesium stearate,
talc, etc. may be used in addition to simple excipients.
Furthermore, the pharmaceutical composition may be formulated into
liquid dosage forms such as suspensions, liquid for internal use,
emulsions, syrups, etc., and various excipients such as humectants,
sweeteners, aromatics, preservatives, etc. in addition to water and
liquid paraffin may be used for the formulation of the liquid
dosage forms.
[0049] For parenteral administration, the pharmaceutical
composition comprising spinosin or the spinosin-containing herbal
extract according to the present invention may include sterile
aqueous solutions, non-aqueous solvents, suspensions, emulsions,
lyophilized preparations, and suppositories. Suitable non-aqueous
solutions and suspensions may include propylene glycol,
polyethylene glycol, vegetable oils such as olive oil, and
injectable esters such as ethyl oleate. Bases for the suppositories
may include Witepsol, macrogol, Tween 61, cacao butter, laurin
butter, glycerogelatine, etc.
[0050] As used herein the term "administration" refers to the
introduction of the composition for prevention and treatment of
cognitive impairments according to the present invention to a
patient in any appropriate way, and the composition for prevention
and treatment of cognitive impairments according to the present
invention may be administered via any conventional administration
route as long as the composition can reach a target tissue. For
example, the composition may be administered orally,
intraperitoneally, intravenously, intramuscularly, subcutaneously,
intradermally, intranasally, intrapulmonary, rectally,
intracavitary, intraperitoneally, or intradurally, but not limited
thereto. For example, the composition may be administered by oral,
rectal, or intravenous, intramuscular, subcutaneous, endometrial,
or intracerebroventricular injection.
[0051] The pharmaceutical composition according to the present
invention may be administered once or several times a day at
regular intervals.
[0052] The pharmaceutical composition according to the present
invention may further comprise other active ingredients effective
for the treatment of the cognitive impairments.
[0053] The pharmaceutical composition according to the present
invention may be used alone or in combination with various methods
such as hormone therapy, drug therapy, etc., for the prevention or
treatment of the cognitive impairments.
[0054] The present invention relates to a food composition
comprising spinosin represented by the above formula 1, a
pharmaceutically acceptable salt thereof, or a spinosin-containing
herbal extract for prevention or improvement of the cognitive
impairments.
[0055] The food composition according to the present invention may
further comprise additives commonly used in food compositions,
health functional foods, or beverages.
[0056] For example, the food composition of the present invention
may comprise sweeteners such as white sugar, crystalline fructose,
glucose, D-sorbitol, mannitol, isomaltooligosaccharide, stevioside,
aspartame, acesulfame potassium, sucralose, etc., acidifiers such
as anhydrous citric acid, DL-malic acid, succinic acid, and salts
thereof, preservatives such as benzoic acid and derivatives
thereof, various nutrients, vitamins, minerals (electrolyte),
flavoring agents such as synthetic and natural flavoring agents,
coloring agents and fillers (cheese, chocolate, etc.), pectic acid
and salts thereof, alginic acid and salts thereof, organic acids,
protective colloidal thickeners, pH-adjusting agents, stabilizers,
preservatives, glycerin, alcohol, carbonators used in carbonated
drinks, etc. Moreover, the food composition of the present
invention may comprise fruit pulp for preparation of natural fruit
juices and vegetable juices. This additive may be used in an amount
of less than about 20 parts by weight with respect to 100 parts by
weight of the food composition.
[0057] When the food composition of the present invention is a
beverage, it may further comprise flavoring agents or natural
carbohydrates. Suitable natural carbohydrates may include
monosaccharides such as glucose and fructose, disaccharides such as
maltose and sucrose, polysaccharides such as dextrin and
cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and
erythritol. Moreover, suitable flavoring agents may include natural
flavoring agents such as thaumatin and stevia extracts
(rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents
such as saccharin and aspartame. When the food composition is a
beverage, the natural carbohydrate may be contained in an amount of
about 1 to 20 g, preferably about 5 to 12 g with respect to 100 mL
of the composition.
[0058] The food composition comprising spinosin or the
spinosin-containing herbal extract according to the present
invention may be prepared in the form of powders, granules,
tablets, capsules, or beverages and used as foods, beverages, gums,
teas, vitamin complexes, health supplement foods, etc.
[0059] The composition comprising spinosin or the
spinosin-containing herbal extract according to the present
invention may be added to drugs, foods, beverages, etc. for
prevention and treatment of the cognitive impairments. For example,
the composition comprising spinosin or the spinosin-containing
herbal extract according to the present invention may be added to
food, beverages, gums, teas, vitamin complexes, health supplement
foods, etc.
[0060] The food composition comprising spinosin or the
spinosin-containing herbal extract according to the present
invention may be added to foods or beverages for prevention or
improvement of the cognitive impairments. The composition of the
present invention may be added in an amount of 1 to 5 wt % with
respect to the total weight of the food and may be added in an
amount of 0.02 to 10 g, preferably 0.3 g to 1 g, with respect to
100 mL of the beverage.
[0061] The present invention provides a pharmaceutical composition
comprising spinosin for prevention or treatment of a degenerative
brain diseases. The spinosin represented by the above formula 1 can
effectively prevent or treat cerebral infarction, stroke,
Parkinson's disease, or Huntington's disease.
[0062] The present invention provides a pharmaceutical composition
comprising a spinosin-containing herbal extract for prevention or
treatment of degenerative brain diseases.
[0063] The spinosin-containing herbal extract can effectively
prevent or treat cerebral infarction, stroke, Parkinson's disease,
or Huntington's disease. The herbal extract may be obtained from
Zizyphus jujuba Mill var. inermis, Zizyphus jujuba Mill var.
hoonensis, Zizyphus jujuba Mill var. spinosa, Passiflora edulis
flavicarpa, Cayaponia tayuya, Desmodium tortuosum, Wilbrandia
ebracteata, Strophioblachia fimbricalyx, Clutia abyssinica,
Saccharopolyspora spinosa, or mixtures thereof, and may preferably
be obtained from the seeds of Zizyphus jujuba Mill var.
spinosa.
[0064] The present invention provides a method for preventing or
treatment of cognitive impairments or degenerative brain diseases,
the method comprising administering to a subject in need thereof a
composition comprising spinosin or a pharmaceutically acceptable
salt thereof. The cognitive impairments may be dementia or amnesia,
and the degenerative brain diseases may be cerebral infarction,
stroke, Parkinson's disease, or Huntington's disease.
[0065] The present invention provides a method for preventing or
treatment of cognitive impairments or degenerative brain diseases,
the method comprising administering to a subject in need thereof a
composition comprising a spinosin-containing herbal extract. The
herbal extract may be obtained from Zizyphus jujuba Mill var.
inermis, Zizyphus jujuba Mill var. hoonensis, Zizyphus jujuba Mill
var. spinosa, Passiflora edulis flavicarpa, Cayaponia tayuya,
Desmodium tortuosum, Wilbrandia ebracteata, Strophioblachia
fimbricalyx, Clutia abyssinica, Saccharopolyspora spinosa, or
mixtures thereof, and may preferably be obtained from the seeds of
Zizyphus jujuba Mill var. spinosa.
Advantageous Effects
[0066] The pharmaceutical composition comprising spinosin or a
spinosin-containing herbal extract according to the present
invention can effectively prevent or treat cognitive impairments.
Specifically, the pharmaceutical composition comprising spinosin or
the spinosin-containing herbal extract according to the present
invention can significantly improve memory and learning ability,
thus effectively prevent or treat dementia.
DESCRIPTION OF DRAWINGS
[0067] FIG. 1 shows the effect of a pharmaceutical composition
comprising spinosin according to the present invention on the
improvement of memory and learning ability.
[0068] FIGS. 2 to 5 are shows the effect of a pharmaceutical
composition comprising a spinosin-containing zizyphi spinosi semen
extract according to the present invention on the improvement of
memory and learning ability.
MODE FOR INVENTION
[0069] Hereinafter, the present invention will be described in
detail with reference to the following Examples and Experimental
Examples. However, the following Examples and Experimental Examples
are merely illustrative of the present invention and the present
invention is not limited by the following Examples and Experimental
Examples.
[0070] Moreover, all reagents and solvents mentioned below were
purchased from Sigma unless otherwise stated, and optical rotations
were measured using a JASCO P-1020 polarimeter. UV spectra were
measured using a Hitachi JP/U3010, IR spectra were measured using a
JASCO FT/IR-5300, NMR spectra was measured using a Bruker Avance
400 (400 MHz), and FAB Mass spectra were measured using a JEOL
JMS-700 mass spectrometer.
Example 1
Preparation of Spinosin-Containing Zizyphi Spinosi Semen
Extract
[0071] 12 kg of zizyphi spinosi semen (the seeds of Ziziphus jujuba
Mill var. spinosa Hu ex H. F. Chou) was crushed by a crusher and
distributed into extraction bottles, n-hexane was added to the
crushed zizyphi spinosi semen until it covered the surface of the
crushed zizyphi spinosi semen, and the mixture was left to stand at
room temperature for 3 days and then filtered. The hexane extracts
were combined and concentrated when the hexane solutions were
transparent after the same method was repeated four times.
[0072] After the removal of the hexane extracts, 70% ethanol was
added to the remaining zizyphi spinosi semen until it covered the
surface of zizyphi spinosi semen, and the mixture was left to stand
at room temperature for 3 days and then filtered and concentrated
under reduced pressure. 70% (v/v) ethanol was added to the residue
and then extracted in water bath, followed by filtration and
concentration. The extraction was performed 6 times by the method
of adding 70% (V/v) ethanol and then extraction in water bath and
filtration, and all the extracts were combined and concentrated
under reduced pressure, yielding 70% (v/v) ethanol soft ext. of
zizyphi spinosi semen.
Example 2
Isolation and Purification of Spinosin
[0073] n-hexane, methanol, and water were added at a volume ratio
of 10:9:1 to the ethanol soft ext. of zizyphi spinosi semen
prepared in the above Example 1, the mixture was shaken and left to
stand overnight to remove the hexane soluble fraction, and then the
90% methanol fraction was concentrated under reduced pressure.
Distilled water was added to the concentrated 90% (v/v) methanol
fraction, ethyl acetate in an amount equivalent to the distilled
water was added, and then the ethyl acetate fraction was removed.
n-butanol of an equivalent amount was added to the water layer, and
then the n-butanol fraction was concentrated under reduced
pressure.
[0074] The n-butanol fraction obtained by concentration under
reduced pressure was loaded onto silica gel column and subjected to
chromatography
(7:1:0.5.fwdarw.7:1.5:0.5.fwdarw.7:2:0.5.fwdarw.MeOH) with
solutions of methylene chloride, methanol, and water at volume
ratios of 7:1:0.5, 7:1.5:0.5, and 7:2:0.5 and methanol as an eluent
to obtain thirty small fractions, and the 25.sup.th small fraction
containing spinosin was obtained from the thirty small fractions by
performing TLC.
[0075] The 25.sup.th small fraction was loaded onto silica gel
column and subjected to chromatography
(100:8:6.fwdarw.100:10:7.5.fwdarw.100:12:9) with solutions of ethyl
acetate, methanol, and water at volume ratios of 100:8:6,
100:10:7.5, 100:12:9 as an eluent to obtain small fractions, and
the 3.sup.rd small fraction containing spinosin was obtained from
the small fractions by performing TLC.
[0076] The 3.sup.rd small fraction was recrystallized with methanol
to yield spinosin (3.2 g) as a pale yellow amorphous powder (yield:
0.027%).
[0077] The physicochemical properties of the spinosin were as
follows:
[0078] TLC Rf: 0.27 (absorbent: silica gel GF.sub.254, developing
solvent: chloroform/methanol/water (520:280:80), color reagent: 20%
sulfuric acid solution)
[0079] Melting point: 237-240.degree. C.
[0080] [.alpha.].sub.D.sup.24=-47.5.degree. (c=0.01, MeOH)
[0081] UV, .lamda..sub.max (log .epsilon.) (MeOH) 216 (sh, 4.74),
272 (4.46), 334 (4.52) nm
[0082] IR, u.sub.max 3141 (OH), 1649, 1603, 1484, 1453, 1347, 1196,
1054, 1020, 841 cm.sup.-1
[0083] .sup.1H-NMR (400 MHz, DMSO-d.sub.6+D.sub.2O) .delta.: 2.56
(1H, dt, J=9.0 Hz, H-5'''), 2.74 (1H, dt, J=9.4 Hz, H-5'''), 2.83
(2H, t, J=8.3, H-2'''), 2.93 (2H, br d, 9.4 Hz, H-6'''), 2.94, 2.99
(1H each, t, J=8.5 Hz, H-4'''), 3.39 (2H, br d, J=12.1 Hz, H-6''),
3.70 (2H, t, J=9.0 Hz, H-3'''), 3.88 (6H, s, OCH.sub.3), 4.15 (2H,
d, J=7.8, H-1'''), 4.28, 4.45 (1H each, t, J=9.3 Hz, H-2''), 4.68
(2H, d, J=9.8 Hz, H-1''), 6.77, 6.79 (1H each, s, H-8), 6.80 (2H,
s, H-3), 6.94 (4H, d, J=8.6 Hz, H-3',5'), 7.95 (4H, d, J=8.6 Hz,
H-2',6'), 13.5, 13.6 (1H each, s, 5-OH)
[0084] .sup.13C-NMR (100 MHz, DMSO-d.sub.6+D.sub.2O) .delta.: 56.5,
56.9 (OCH.sub.3), 60.2, 60.8 (C-6'''), 61.6 (C-6''), 69.3, 69.6
(C-4'''), 70.6 (C-4''), 71.1, 71.4 (C-1''), 74.7, 74.8 (0-2'''),
76.4 (C-3'''), 76.6, 76.8 (C-5'''), 78.4, 78.7 (C-3''), 80.9, 81.3
(C-2''), 81.7, 82.0 (C-5''), 90.8, 91.3 (C-8), 103.4, 103.5 (C-3),
104.5, 104.7 (C-10), 105.4, 105.6 (C-1'''), 108.8 (C-6), 116.4
(C-3',5'), 121.4, 121.5 (C-1'), 128.9 (C-2',6'), 157.4, 157.5
(C-9), 159.6, 160.5 (C-5), 161.4 (0-4'), 164.2 (C-2), 164.3, 165.4
(C-7), 182.3, 182.6 (C-4)
[0085] Positive FAB-MS m/z 609 [M+H].sup.+
Experimental Example
[0086] Determination of Effect on Dementia Treatment
[0087] Experiments were performed using models with memory
impairment induced by scopolamine to determine the effect of the
spinosin and the zizyphi spinosi semen extract containing spinosin
as an active ingredient prepared in Examples 1 and 2, and the
detailed experimental methods are as follows:
1) Preparation of Experimental Animals
[0088] 6-week-old ICR mice (Orient Bio Inc., Korea), weighing about
26 to 28 g, were fed with water and food freely available for 5
days in an environment, where the temperature was about
23.+-.2.degree. C., the humidity was about 55.+-.10%, and the
light-dark cycle was 12 hours (Animal Laboratory at the College of
Pharmacy in Kyung Hee university), and used in the experiments.
2) Statistical Analysis
[0089] Statistical analysis of all experimental data was performed
using a one-way analysis of variance (ANOVA), and when the
statistical significance was recognized, the significance was
assessed at P<0.05 using Student-Newman-Keuls Test.
3) Experimental Example 1
Passive Avoidance Test 1
[0090] A passive avoidance test apparatus was prepared for the
experiment. The passive avoidance test apparatus was divided into a
first space and a second space, a guillotine door was placed
between the two spaces, and the first and second spaces are
connected by the door. The first space was kept light using
illumination, and the second space was kept dark. The floor of the
second space kept dark was provided with a grid, and an electric
shock of 0.5 mA was applied for 3 seconds through the grid on the
floor when the experimental animal moved to the dark space.
[0091] The mice prepared in the above section 1) were divided into
7 groups including drug administration groups 1 to 4, to which
spinosin was to be administered, and control groups 1 to 3 (10 mice
per group). The spinosin of Example 2 was dissolved in 10% Tween 80
(Polyoxyethylene sorbitan monooleate: Sigma, U.S.A.) and
administered to drug administration groups 1 to 4 at doses of 2.5
mg/Kg, 5 mg/Kg, 10 mg/Kg, and 20 mg/Kg. Meanwhile, donepezil
(Sigma-Aldrich Chemistry Co.) was administered to control group 1
at a dose of 5 mg/Kg, and 10% Tween 80 was administered to control
groups 2 and 3 at a dose of 0.15 mL.
[0092] After about 30 minutes, scopolamine (Sigma-Aldrich Chemistry
Co.) dissolved in distilled water was intraperitoneally
administered to drug administration groups 1 to 4 and control
groups 1 and 2 at a dose of 1 mg/Kg (Ebert, U. et al., Eur. J.
Clin. Invest., 28, pp 944-949, 1998), and 0.9% saline solution was
intraperitoneally administered to control group 3. After 30
minutes, learning about the passive avoidance test apparatus was
performed on the mice in drug administration groups 1 to 4 and
control groups 1 to 3, respectively. Specifically, the mice were
placed in the first space kept light, and after a seek time of
about 20 seconds, the guillotine door was opened, and the latency
time taken for the mice to move to the second space kept dark was
measured.
[0093] The mice that did not move to the second space kept dark
until 60 seconds elapsed after the guillotine door was opened were
excluded from the experiment.
[0094] After 24 hours since the learning was completed, the present
experiment was performed on drug administration groups 1 to 4 and
control groups 1 to 3, respectively. The latency time taken for all
four feet of the mouse to enter the dark space after the guillotine
door was opened from the seek time of 10 seconds was measured up to
300 seconds. A longer latency time represents that the learning and
memory of the passive avoidance is better.
[0095] In learning and in the present experiment, the average
latency times of the mice in each group are shown in the following
Table 1 and FIG. 1:
TABLE-US-00001 TABLE 1 Present Experiment Experimental Group
Learning (sec) (sec) Drug administration 25.88 .+-. 13.72 59.13
.+-. 46.88 group 1 (2.5 mg/Kg) Drug administration 29.38 .+-. 18.17
107.1 .+-. 91.70 group 2 (5 mg/Kg) Drug administration 20.50 .+-.
9.196 131.3 .+-. 69.94 group 3 (10 mg/Kg) Drug administration 36.00
.+-. 16.45 146.0 .+-. 83.53 group 4 (20 mg/Kg) Control group 1
25.13 .+-. 13.68 151.9 .+-. 37.24 (donepezil) Control group 2 26.50
.+-. 18.16 41.33 .+-. 18.92 Control group 3 29.17 .+-. 18.15
300
[0096] As can be seen from Table 1 and FIG. 1, the latency time
taken for the mice in drug administration groups 1 to 4, to which
the spinosin prepared in Example 2 was administered, to move the
second space was significantly increased compared to that in
control group 2 to which only scopolamine was administered.
Specifically, the latency time in drug administration group 1, to
which the spinosin was administered at a dose of 2.5 mg/Kg, was
increased about 20 seconds compared to that in control group 2, and
the latency time in drug administration group 2, to which the
spinosin was administered at a dose of 5 mg/Kg, was increased about
2.5 times compared to that in control group 2. Moreover, the
latency time in drug administration group 3, to which the spinosin
was administered at a dose of 10 mg/Kg, was increased about 3
times, compared to that in control group 2, and the latency time in
drug administration group 4, to which the spinosin was administered
at a dose of 20 mg/Kg, was increased about 3.5 times compared to
that in control group 2. Particularly, it could be seen that the
memory and learning ability of the mice with memory impairment
induced by scopolamine in drug administration groups 3 and 4 was
improved to an extent similar to that in control group 1 to which
donepezil was administered.
[0097] It can be seen from these results that the spinosin prepared
in Example 2 can effectively prevent or treat cognitive impairments
such as dementia.
4) Experimental Example 2
Passive Avoidance Teat 2
[0098] The passive avoidance test was performed on drug
administration groups 1 to 4 and control groups 1 to 3 by the same
method as Experimental Example 1, except that the zizyphi spinosi
semen extract extracted with 70% ethanol in Example 1 was dissolved
in 10% Tween 80 and administered to drug administration groups 1 to
4 at doses of 25 mg/kg, 50 mg/kg, 100 mg/kg, and 200 mg/kg.
[0099] The results of the experiment are shown in the following
Table 2 and FIG. 2:
TABLE-US-00002 TABLE 2 Present Experiment Experimental Group
Learning (sec) (sec) Drug administration 25.75 .+-. 15.92 49.50
.+-. 46.26 group 1 (25 mg/Kg) Drug administration 23.25 .+-. 18.17
40.00 .+-. 17.51 group 2 (50 mg/Kg) Drug administration 32.13 .+-.
18.92 173.9 .+-. 77.84 group 3 (100 mg/Kg) Drug administration
27.75 .+-. 19.00 112.8 .+-. 89.67 group 4 (200 mg/Kg) Control group
1 17.25 .+-. 5.365 140.5 .+-. 25.47 (donepezil) Control group 2
32.80 .+-. 21.26 26.10 .+-. 25.27 Control group 3 21.00 .+-. 8.380
228.7.00 .+-. 50.65
[0100] As can be seen from Table 2 and FIG. 2, the latency time
taken for the mice in drug administration groups 1 to 4, to which
the spinosin-containing zizyphi spinosi semen extract prepared in
Example 1 was administered, to move the second space was
significantly increased compared to that in control group 2 to
which only scopolamine was administered. Specifically, the latency
time in drug administration groups 1 and 2, to which the spinosin
was administered at doses of 25 mg/Kg and 50 mg/Kg, respectively,
was increased about 2 times compared to that in control group 2.
Moreover, the latency time in drug administration group 3, to which
the spinosin was administered at a dose of 100 mg/Kg, was increased
about 6 times compared to that in control group 2, and the latency
time in drug administration group 4, to which the spinosin was
administered at a dose of 200 mg/Kg, was increased about 4.3 times
compared to that in control group 2. Particularly, it could be seen
that the memory and learning ability of the mice with memory
impairment induced by scopolamine in drug administration groups 3
and 4 was improved to an extent similar to that in control group 1
to which donepezil was administered.
[0101] It can be seen from these results that the
spinosin-containing zizyphi spinosi semen extract prepared in
Example 1 can effectively prevent or treat cognitive impairments
such as dementia.
5) Experimental Example 3
Y-Maze Test
[0102] A Y-maze was prepared for the experiment. The Y-maze had
three arms, in which each arm was 42 cm in length, 3 cm in width,
and 12 cm in height, the angle formed by three arms was 120
degrees, and the Y-maze was made of black polyvinyl resin.
[0103] The mice prepared in the above section 1) were divided into
5 groups including drug administration groups 1 and 2, to which the
zizyphi spinosi semen extract prepared in Example 1 was to be
administered, and control groups 1 to 3 (10 mice per group).
[0104] 160 mg of the zizyphi spinosi semen extract of Example 1 was
dissolved in 4 mL of 10% Tween 80 (Polyoxyethylene sorbitan
monooleate: Sigma, U.S.A.) and administered to drug administration
groups 1 and 2 at doses of 100 mg/Kg and 200 mg/Kg. Meanwhile,
donepezil (Sigma-Aldrich Chemistry Co.) was administered to control
group 1 at a dose of 5 mg/Kg, and 10% Tween 80 was administered to
control groups 2 and 3 at a dose of 0.15 mL.
[0105] After about 30 minutes, scopolamine (Sigma-Aldrich Chemistry
Co.) dissolved in distilled water was intraperitoneally
administered to drug administration groups 1 and 2 and control
groups 1 and 2 at a dose of 1 mg/Kg (Ebert, U. et al., Eur. J.
Clin. Invest., 28, pp 944-949, 1998), and 0.9% saline solution was
intraperitoneally administered to control group 3.
[0106] After 30 minutes, the mice in drug administration groups 1
and 2 and control groups 1 to 3 were carefully placed on one of
three arms of the Y-maze, divided into A, B, and C, respectively,
and allowed to freely move, and then the arms where the mice
entered were recorded. At this time, only the case where the mouse'
tail fully entered the arm was recorded as the arm where the
experimental animal entered, and the case where the mouse entered
the same arm was also recorded.
[0107] 1 point was given to the case where the mouse sequentially
entered three different arms (actual alternation). Alternation
behavior was defined as the case where the mouse entered all three
arms and was converted into a percentage (%) by the following
Formula 1 (Sarter, M. et al., Psychopharmacology., 94, pp 491-495,
1998).
Alternatoin behavior ( % ) = Actual alternation Maximum alternation
.times. 100 [ Formula 1 ] ##EQU00001##
(Maximum alternation: Total entry--2)
[0108] The alternation behaviors converted by the above Formula 1
are shown in the following Table 3 and FIG. 3, and the total
entries, each representing the total number of entries, are shown
in the following Table 4 and FIG. 4:
TABLE-US-00003 TABLE 3 Experimental Group Alternation behavior (%)
Drug administration group 1 63.32 .+-. 9.005 (100 mg/Kg) Drug
administration group 2 60.91 .+-. 6.369 (200 mg/Kg) Control group 1
(Donepezil) 65.71 .+-. 10.19 Control group 2 50.92 .+-. 14.59
Control group 3 76.53 .+-. 8.379
TABLE-US-00004 TABLE 4 Experimental Group Total entry Drug
administration group 1 37.00 .+-. 14.30 (100 mg/Kg) Drug
administration group 2 32.30 .+-. 7.675 (200 mg/Kg) Control group 1
(Donepezil) 34.75 .+-. 7.265 Control group 2 24.75 .+-. 6.519
Control group 3 26.88 .+-. 13.78
[0109] As can be seen from Table 3 and FIG. 3, the alternation
behaviors in drug administration groups 1 and 2, to which the
spinosin-containing zizyphi spinosi semen extract prepared in
Example 1 was administered, were increased compared to those in
control group 2, to which only scopolamine was administered, and
were similar to those in control group 1 to which donepezil was
administered. Moreover, as can be seen from Table 4 and FIG. 4, the
total entries in drug administration groups 1 and 2 and control
groups 1 to 3 were all similar to each other. Therefore, it could
be seen that the increase in the alternation behavior did not
result from the change in the activity of the mice, and thus it
could be seen that the memory and learning ability of the mice in
drug administration groups 1 and 2 was significantly improved by
the spinosin-containing zizyphi spinosi semen extract prepared in
Example 1.
[0110] It can be seen from these results that the
spinosin-containing zizyphi spinosi semen extract prepared in
Example 1 can effectively prevent or treat cognitive impairments
such as dementia.
6) Experimental Example 4
Morris Water Maze Test
[0111] A Morris water maze test apparatus was prepared for the
experiment. Four labels such as star, square, triangle, and circle
were put at regular intervals on a circular pool of 90 cm in
diameter and 45 cm in height, and a platform of 29 cm in height was
positioned below the star. Water was filled up to 0.5 cm from the
platform (water temperature: 21.+-.1.degree. C.) and became cloudy
with pigment such that the platform was not visible from the
surface of the water.
[0112] The mice prepared in the above section 1) were divided into
5 groups including drug administration groups 1 and 2, to which the
zizyphi spinosi semen extract prepared in Example 1 was to be
administered, and control groups 1 to 3 (10 mice per group).
[0113] 160 mg of the zizyphi spinosi semen extract of Example 1 was
dissolved in 4 mL of 10% Tween 80 (Polyoxyethylene sorbitan
monooleate: Sigma, U.S.A.) and administered to drug administration
groups 1 and 2 at doses of 100 mg/Kg and 200 mg/Kg. Meanwhile,
donepezil (Sigma-Aldrich Chemistry Co.) was administered to control
group 1 at a dose of 5 mg/Kg, and 10% Tween 80 was administered to
control groups 2 and 3 at a dose of 0.15 mL.
[0114] After about 30 minutes, scopolamine (Sigma-Aldrich Chemistry
Co.) dissolved in distilled water was intraperitoneally
administered to drug administration groups 1 and 2 and control
groups 1 and 2 at a dose of 1 mg/Kg (Ebert, U. et al., Eur. J.
Clin. Invest., 28, pp 944-949, 1998), and 0.9% saline solution was
intraperitoneally administered to control group 3.
[0115] After 30 minutes, the time taken for the mice, put down in
one section of the pool, in drug administration groups 1 and 2 and
control groups 1 to 3 to reach the platform was measured for 60
seconds. After 30 minutes, the time taken for the mice,
repositioned to the original position, to reach the platform was
measured until 60 seconds. The above-described process was repeated
4 times, and the average values were obtained. The experiment was
performed in the same manner as on the first day while
administering the drugs for 4 days, except that the position where
the mice were first put down was changed.
[0116] The results of the experiment are shown in the following
Table 5 and FIG. 5:
TABLE-US-00005 TABLE 5 Experimental Time (sec) group 1.sup.st day
2.sup.nd day 3.sup.rd day 4.sup.th day Drug 56.10 .+-. 9.088 59.85
.+-. 0.4743 49.85 .+-. 16.64 46.70 .+-. 15.96 administration group
1 (100 mg/Kg) Drug 52.75 .+-. 12.17 46.20 .+-. 12.52 43.45 .+-.
18.19 30.55 .+-. 20.45 administration group 2 (200 mg/Kg) Control
group 1 59.34 .+-. 2.055 48.75 .+-. 14.98 42.80 .+-. 15.16 34.35
.+-. 13.80 (Donepezil) Control group 2 57.75 .+-. 7.115 57.05 .+-.
6.693 53.95 .+-. 11.13 57.30 .+-. 6.701 Control group 3 57.10 .+-.
6.736 40.35 .+-. 14.98 27.65 .+-. 16.27 25.60 .+-. 13.70
[0117] As can be seen from Table 5 and FIG. 5, the time taken for
the mice in drug administration groups 1 and 2, to which the
spinosin-containing zizyphi spinosi semen extract prepared in
Example 1 was administered, to reach the platform was reduced by
more than 10 seconds and 25 seconds, respectively, on the 3.sup.rd
and 4.sup.th day compared to that in control group 2 to which only
scopolamine was administered. Particularly, in the case of drug
administration group 2, the time taken for the mice to reach the
platform was reduced compared to that in control group 1 to which
donepezil was administered, indicating that the spinosin-containing
zizyphi spinosi semen extract prepared in Example 1 had superior
memory and learning ability to the donepezil.
[0118] Preparation of Formulations and Food Compositions
[0119] Preparation of Powders
[0120] Powders were prepared by mixing 20 mg of the
spinosin-containing zizyphi spinosi semen extract prepared in
Example 1, 100 mg of lactose, and 10 mg of talc and packing the
mixture in airtight bags.
[0121] Preparation of Tablets
[0122] Tablets were prepared using the spinosin-containing zizyphi
spinosi semen extract prepared in Example 1 and the ingredients
shown in the following Table 6:
TABLE-US-00006 TABLE 6 Ingredients Content (mg) Zizyphi spinosi
semen extract 10 (Example 1) Corn starch 100 Lactose 100 Magnesium
stearate 2
[0123] Tablets were prepared by mixing the above ingredients and
compressing the mixture into tablets according to a conventional
method for preparing tablets.
[0124] Preparation of Capsules
TABLE-US-00007 TABLE 7 Ingredients Content (mg) Zizyphi spinosi
semen extract 10 (Example 1) Crystalline cellulose 100 Lactose 100
Magnesium stearate 0.2
[0125] Capsules were prepared by mixing the above ingredients and
filling the mixture in gelatin capsules according to a conventional
method for preparing capsules.
[0126] Preparation of Injections
TABLE-US-00008 TABLE 8 Ingredients Content (mg) Zizyphi spinosi
semen extract 10 (Example 1) Mannitol 180 Sterile distilled water
for 2974 injection Na.sub.2HPO.sub.4.cndot.12H.sub.2O 26
[0127] Injections were prepared by mixing the above ingredients to
prepare a solution and filling the solution in ampoules (2 mL),
followed by sterilization, according to a conventional method for
preparing injections.
[0128] Preparation of Liquid Formulations
TABLE-US-00009 TABLE 9 Ingredients Content Zizyphi spinosi semen
extract 20 mg (Example 1) Isomerized sugar 10 g Mannitol 5 g
Purified water 85 ml
[0129] Liquid formulations were prepared by dissolving the above
ingredients in purified water, adding a suitable amount of lemon
flavor, adding purified water to the mixture to prepare a solution
of 100 ml, and loading the solution into brown bottles, followed by
sterilization, according to a conventional method for preparing
liquid formulations.
[0130] Preparation of Health Foods
TABLE-US-00010 TABLE 10 Ingredients Content Zizyphi spinosi semen
extract 1000 mg (Example 1) Nicotinic acid amide 1.7 mg Folic acid
50 .mu.g Calcium pantothenate 0.5 mg Ferrous sulfate 1.75 mg Zinc
oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium Phosphate
monobasic 15 mg calcium Phosphate dibasic 55 mg Potassium citrate
90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg Vitamin
mixture Suitable amount Mineral mixture Suitable amount
[0131] Health food compositions were prepared according to a
conventional method by mixing the above ingredients according to a
conventional method for preparing health foods and then preparing
granules.
[0132] Preparation of Health Beverages
TABLE-US-00011 TABLE 11 Ingredients Content Zizyphi spinosi semen
extract 1000 mg (Example 1) Citric acid 1000 mg Oligosaccharide 100
g Japanese apricot concentrate 2 g Taurine 1 g Purified water Total
900 mL
[0133] Health beverages were prepared by dissolving the above
ingredients in purified water to prepare a solution of a 900 mL,
stirring and heating the solution at 85.degree. C. for about 1
hour, filtering the solution, filling the solution in sterilized 2
L bottles, sealing and sterilizing the bottles, and keeping the
bottles under refrigeration according to a conventional method for
preparing health beverages.
* * * * *