U.S. patent application number 14/361012 was filed with the patent office on 2014-10-30 for composition comprising vicenin-2 having a beneficial effect on neurological and/or cognitive function.
The applicant listed for this patent is AMINO UP CHEMICAL CO., LTD.. Invention is credited to Sybille Buchwald-Werner, Hajime Fujii.
Application Number | 20140322198 14/361012 |
Document ID | / |
Family ID | 48534700 |
Filed Date | 2014-10-30 |
United States Patent
Application |
20140322198 |
Kind Code |
A1 |
Buchwald-Werner; Sybille ;
et al. |
October 30, 2014 |
COMPOSITION COMPRISING VICENIN-2 HAVING A BENEFICIAL EFFECT ON
NEUROLOGICAL AND/OR COGNITIVE FUNCTION
Abstract
The invention relates to an active ingredient and/or to a
composition having beneficial effect on neurological and cognitive
functions.
Inventors: |
Buchwald-Werner; Sybille;
(Dusseldorf, DE) ; Fujii; Hajime; (Ebetsu,
JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
AMINO UP CHEMICAL CO., LTD. |
Sapporo |
|
JP |
|
|
Family ID: |
48534700 |
Appl. No.: |
14/361012 |
Filed: |
November 29, 2012 |
PCT Filed: |
November 29, 2012 |
PCT NO: |
PCT/EP2012/074010 |
371 Date: |
May 28, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61564374 |
Nov 29, 2011 |
|
|
|
61622260 |
Apr 10, 2012 |
|
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Current U.S.
Class: |
424/130.1 ;
424/184.1; 424/641; 424/725; 424/728; 424/730; 424/734; 424/745;
424/746; 424/752; 424/769; 424/94.1; 424/94.67; 514/23;
536/1.11 |
Current CPC
Class: |
A61K 31/7048 20130101;
A61P 25/22 20180101; A61P 25/14 20180101; A61K 36/00 20130101; A61P
25/20 20180101; A61K 31/7048 20130101; A61K 36/535 20130101; A61P
25/28 20180101; A61K 45/06 20130101; A61P 25/24 20180101; A61P
25/16 20180101; A61K 2300/00 20130101 |
Class at
Publication: |
424/130.1 ;
514/23; 424/745; 424/184.1; 424/94.1; 424/641; 424/746; 424/728;
424/752; 424/734; 424/730; 424/725; 424/769; 424/94.67;
536/1.11 |
International
Class: |
A61K 31/7048 20060101
A61K031/7048; A61K 45/06 20060101 A61K045/06; A61K 36/535 20060101
A61K036/535 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 29, 2011 |
EP |
11009430.7 |
Apr 10, 2012 |
EP |
12163578.3 |
Claims
1. A method of maintaining and/or improving neurological and/or
brain function and/or preventing, delaying onset, controlling
and/or treating a neurological dysfunction, condition, disorder or
disease, the method comprising administering vicenin 2 or a
biologically active analogue thereof as an active ingredient to a
subject in need thereof.
2. The method of claim 1, for maintaining and/or improving
neurological and/or brain function.
3. The method of claim 1, wherein the neurological dysfunction,
condition, disorder or disease is an age or stress related
neurological dysfunction, anxiety, a cognitive disorder, depression
or dementia.
4. The method of claim 1, wherein the active ingredient is derived
from a plant.
5. The method of claim 4, wherein the active ingredient is
comprised in a plant preparation enriched for the active
ingredient.
6. The method of claim 4, wherein the plant is selected of one or
more plants from a group consisting of a Anethum, Perilla, Urtica,
Passiflora, Camelia, Cayaponia, Colocasia, Cydonia, Desmodium,
Hordeum, Origanum, Ocimum, Jatropha, Parkinsonia, Peperomia,
Piheranthos, Centaurea, Indigo, Bomba, Lychnophera, Asplenium,
Chinotto, Citrus, Viola, Trigonella, Rosemary, Peppermint, Thyme,
Basil, Sage, Oregano, Lavandula, Nipponanthemum, Abrus, Viola,
Santalum, Oryza, Scleropyrum, Tulsi, Centaurea, Indigofera, Bombax,
Glinus, Lychnophora, other species belonging to the Lamiacea,
Labiatae, and Urticaceae, Rosales or Malpighiales or a combination
thereof.
7. The method of claim 1, wherein the active ingredient is derived
from a plant preparation selected from the group consisting of a
leaf preparation, a fruit preparation, a seed preparation, a stem
preparation, a flower preparation, a bud preparation, a root
preparation and a mixture of different parts of the plant.
8. The method of claim 1, wherein the active ingredient is an
isolated vicenin 2 or biologically active analogue thereof obtained
by isolation or chemical synthesis.
9. The method of claim 1, wherein the neurological and/or brain
function is selected from the group consisting of memory,
attention, concentration, alertness, mental flexibility and/or
speed, learning, intelligence, language skills, problem solving
capacity, consciousness, coping with psychological stress or
tension, motivation, mobility, decision making capacity, reaction
time and regulation of emotions.
10. The method of claim 1, wherein the active ingredient is
comprised in a composition and the composition is a food product, a
dietary supplement or a medicament.
11. The method of claim 10, wherein the concentration of the active
ingredient is from 0.1 .mu.g to 500 .mu.g.
12. The method of claim 10, wherein the composition is
substantially free of flavonoids of other plants, other plant
extracts or other plant flavonoids.
13. The method of claim 10, further comprising an agent other than
vicenin 2 or the biologically active analogue thereof capable of
maintaining and/or improving neurological and/or brain function
and/or preventing, delaying onset, controlling and/or treating a
neurological dysfunction, condition, disorder or disease.
14. The method of claim 13, wherein the agent is a lipid, a lipid
containing omega-3-fatty acid, physiological active fatty acid, an
antioxidant, an anti-inflammatory agent, a bulking agent, an immune
system modulatory agent or a vaccine, an antibody, a metal-protein
interaction attenuation agent, a plant preparation, curcumin,
coenzym Q10, L-carnitine, zinc, epigallocatechingallate, thymol,
p-cymere, vinpocetine, hyperzine A, phosphatidylserine, a vitamin,
alpha liponic acid, a TNF alpha inhibitor, a flavonoid, an
anthocyanidin, a biflavonoid, a flavon, a flavonglycoside or a
carboxylic acid.
15. The method of claim 14, wherein the plant preparation is
selected from one or more extracts from a group consisting of an
extract of Ginkgo biloba, Hypericum perforatum, Hyperzia serrata,
Galanthus nivalis, Salvia officinalis, Panex ginseng, Lippia
citriodora, Melissa officinalis, Passiflora incarnate, Passiflora
edulis, Bacopa monniera, Zingiber officinalis, Leucojum aestrum,
Concolulus pluricaulis, Centella asiatica, Emblica officinalis,
Coptidis Rhizoma, Salvia triloba, Piper nigrum, Trigonella
foenum-graecum, Cimicifuga racemosa, Salvia miltiorrhiza, Rhodiola
rosea, Habranthus jamesonii, Phycella herbertiana, Rhodophiala
mendocina, Zephyranthes filifolia, Stephania pierrei, Kaempfera
parviflora, Stephania venosa, Crocus sativus, Salvia species,
Bacopa monnieri, Centella asiatica, Ptychopetalum olacoides,
Withania somnifera, Coptis chinensis, Mangifera indica, Polygala
caudata, Polygala tenuifolia, Halenia elliptica, Evolvilus
alsinoides, Celastrus paniculatus, Clitoria ternatea, Curcuma
longa, Acorus calamus, Terminalia chebula, Lycoris radiata,
Magnolia officinalis, Biota orientalis, Codonopsis pilosula, Evodia
rutaecarpa, Polygonum multiflorum, Aspalathus linearis, Cyclopia
species, Adansonia digitata, Sclerocarya birrea, Mangifera indica,
Actinidia chinensis and/or Matricaria recutita or combinations
thereof.
16. The method of claim 13, wherein the agent is an
acetylcholinesterase inhibitor, a NMDA (N-methyl-D-aspartate)
receptor inhibitor, a non steroidal anti-rheumatic agent, a
neuroleptic agent, a tricyclic antidepressant, an anti-psychotic
agent, a gab junction inhibitor, a non selective monoaminooxidase
inhibitor, an ABCC1 Transporter, an ADAM 10 protein,
methylthioninium chloride, an antibiotic agent, an antiviral agent,
a gamma secretase inhibitor, a beta secretase inhibitor, an
angiotensin receptor antagonist (AT1 antagonist), a cannabinoid,
allopregnanolone or an insulin sensitizer.
17. A process of producing a preparation of vicenin 2 or a
biologically active analogue thereof as an active ingredient
comprising the steps of: a) selecting a raw plant material, b)
preparing the raw plant material, c) applying an extraction process
using aqueous solvent extraction and/or filtration techniques, d)
determining the concentration of the active ingredient; and e)
selecting the preparation comprising the active ingredient at a
concentration of at least 0.01%.
18. The process of claim 17, wherein the plant is selected from the
group consisting of Anethum, Perilla, Urtica, Passiflora, Camelia,
Cayaponia, Colocasia, Cydonia, Desmodium, Hordeum, Origanum,
Ocimum, Jatropha, Parkinsonia, Peperomia, Piheranthos, Centaurea,
Indigo, Bomba, Lychnophera, Asplenium, Chinotto, Citrus, Viola,
Trigonella, Rosemary, Peppermint, Thyme, Basil, Sage, Oregano,
Lavandula, Nipponanthemum, Abrus, Viola, Santalum, Oryza,
Scleropyrum, Tulsi, Centaurea, Indigofera, Bombax, Glinus,
Lychnophora, other species belonging to the Lamiacea, Labiatae, and
Urticaceae, Rosales or Malpighiales or a combination thereof.
19. A preparation of the active ingredient obtained by the process
of claim 17.
20. (canceled)
21. (canceled)
22. The method of claim 3, wherein the the dementia is Alzheimer
disease.
23. The method of claim 1, wherein the neurological or brain
function is selected from the group consisting of memory,
attention, concentration, alertness, mental flexibility and/or
speed, learning, intelligence, language skills, problem solving
capacity, consciousness, coping with psychological stress or
tension, motivation, mobility, decision making capacity, reaction
time and regulation of emotions.
24. A kit comprising vicenin 2 or a biologically active analogue
thereof as an active ingredient and instructions for administering
said composition.
25. The method of claim 1, wherein the neurological disorder is
associated with autonomous neuronal death by blocking the gap
junction hemichannel of activated microglia and reducing their
glutamate release.
26. The method of claim 25, wherein the disorder is Alzheimer
disease, Parkinson disease, Huntington disease, multiple sclerosis,
amylotrophic lateral sclerosis (ALS), encephalitis or AIDS.
27. The method of claim 25, wherein the composition comprises or is
derived of a plant preparation comprising the active
ingredient.
28. The method of claim 27, wherein the plant preparation is
derived from a plant selected from a group consisting of a Anethum,
Perilla, Urtica, Passiflora, Camelia, Cayaponia, Colocasia,
Cydonia, Desmodium, Hordeum, Origanum, Ocimum, Jatropha,
Parkinsonia, Peperomia, Piheranthos, Centaurea, Indigo, Bomba,
Lychnophera, Asplenium, Chinotto, Citrus, Viola, Trigonella,
Rosemary, Peppermint, Thyme, Basil, Sage, Oregano, Lavandula,
Nipponanthemum, Abrus, Viola, Santalum, Oryza, Scleropyrum, Tulsi,
Centaurea, Indigofera, Bombax, Glinus, Lychnophora, other species
belonging to the Lamiacea, Labiatae, and Urticaceae, Rosales or
Malpighiales or a combination thereof and/or is plant preparation
selected from the group consisting of a leaf preparation, a fruit
preparation, a seed preparation, a stem preparation, a flower
preparation, a bud preparation, a root preparation and a mixture of
different parts of the plant.
29. The method of claim 1, wherein the active ingredient has more
than one effect selected from the group consisting of reversible
acetylcholinesterase inhibition, blockade of gap junction
hemichannels, neuroleptic effect, anti-depressive effect,
enkephaline like effect and reduction in TNF alpha.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a U.S. National Stage application of
PCT/EP2012/074010 filed 29 Nov. 2012, which claims priority to
European Patent Application 11009430.7 filed 29 Nov. 2011, European
Patent Application 12163578.3 filed 10 Apr. 2012, U.S. Provisional
Patent Application 61/564,374 filed 29 Nov. 2011, and U.S.
Provisional Patent Application 61/622,260 filed 10 Apr. 2012, the
entire disclosures of which are hereby incorporated by
reference.
TECHNICAL FIELD
[0002] The invention relates to a food, a dietary supplement and a
drug composition comprising as an active ingredient vicenin 2 or a
biologically active analogue thereof, and especially its beneficial
effects to maintain and/or improve neurological and brain function
and to prevent, delay onset, control and/or treat a cognitive
dysfunction, condition, disorder or disease, e.g. dementia such as
Alzheimer disease.
BACKGROUND OF THE INVENTION
[0003] Maintenance or improvement of neurological and brain
functions is important to vitality and well being throughout all
stages of life but particularly important for elderly people. The
efficiency of each person's neurological and brain functions can
make all the difference in daily and overall health. The ability of
cognitive functions, including memory, attention, concentration,
alertness, mental flexibility and speed, learning, intelligence,
language, problem solving capacity, consciousness, coping with
psychological stress or tension, motivation, mobility, decision
making and reaction time as well as emotions like anxiety and mood
swings affect social and economical status and the overall quality
of life. The wish to extend cognitive capacity longer, frame a
lifelong challenge, as everybody is effected by aging. In addition,
there is an increasing number of individuals, who develop
neurodegenerative diseases, like dementia or Alzheimer disease.
[1,2]
[0004] Dementia can be divided into three types, referring to their
cause of development. The first type is called vascular dementia,
caused by reduced circulation; the second type is called secondary
dementia, generated as a side effect of e.g. hormonal disorders and
the third one, is Alzheimer. Alzheimer disease is the most common
form of dementia, forming up to 60% of the cases. There is no cure
for the disease, which worsens as it progresses, and eventually
leads to death.
[0005] Alzheimer is a devastating neurological disorder that
affects more than 37 million people worldwide. The economic burden
of Alzheimer's disease is massive and currently approved drugs do
not prevent or reverse the disease and provide only modest
symptomatic benefits. [3] The pathogenesis of Alzheimer disease is
not completely understood. Major characteristics of Alzheimer's
disease (AD) are synaptic loss, cholinergic dysfunction, and
abnormal protein depositions in the brain. A combination of several
causes may lead to the development of plaques in the brain, which
decrease the efficiency of neuronal communication leading to death
of neurons. Scientifically these plaques are called "beta amyloid
peptides" or "Tau-proteins". [3,4] Recent studies confirmed that a
genetic disposition may be involved and/or infection may contribute
to the formation of plaques and the development of Alzheimer
disease. The decline in neurological and brain functions is linked
to the reduced number of neurons, which lead to a reduced synthesis
of the neurotransmitter acetylcholine and glutamine. The increase
of neurotransmitter concentration, by inhibiting its metabolism is
a possibility to promote beneficially the neurological and brain
function or to reduce the development of dementia like Alzheimer
disease. [4]
[0006] Not many drugs are approved for the treatment of dementia
such as Alzheimer. Four approved drugs target the increase of
neurotransmitter concentration. Recent drug developments and areas
of clinical research focus on treating the cause of the disease by
reduction of amyloid beta levels. Immunotherapy or vaccinations
with anti-amyloid antibodies are being investigated. Another
substance, called PBT2 binds metals which are necessary for
building of protein structures forming the plaques. Other proteins
may have the ability to cut amyloid beta into small pieces, which
can be eliminated. In addition, gab junction antagonists are being
investigated. They block confused communication between cells to
promote a healthy neurological and brain functionality. [3,4] As of
2012, more than 1000 clinical trials have been or are being
conducted to find ways to treat Alzheimer disease. [1,1] Despite
considerable efforts by academic researchers and pharmaceutical and
food industry, the development of novel ingredients for the
maintenance and improvement of neurological and brain function and
to prevent the development and to slow down the progression of
dementia and Alzheimer disease has been dragging and did not lead
to many innovations. Thus, there is a need to develop drugs,
functional foods and dietary supplement ingredients which may
contribute to the maintenance and improvement of neurological and
brain function and which may prevent the development of cognitive
disorders or slow down the progression of dementia and Alzheimer
disease.
[0007] The object for the present invention was to provide novel
active ingredients suitable as functional food, dietary supplement
ingredients and drugs, which are able to maintain and improve
neurological and brain function and to prevent, delay onset,
control and treat cognitive dysfunctions, conditions, disorders or
diseases, in particular dementia such as Alzheimer disease.
SUMMARY OF THE INVENTION
[0008] In accordance with the present invention it has been
surprisingly found that the active ingredient vicenin 2 or
biologically active analogues thereof and compositions comprising
the same have beneficial effects on cognitive functions including
memory, attention, concentration, alertness, mental flexibility
and/or speed, learning, intelligence, language, problem solving
capacity, consciousness, coping with psychological stress or
tension, motivation, mobility, decision making capacity and
reaction time as well as emotions like anxiety and mood swings.
[0009] Thus, the invention describes a new neurological and
cognitive active agent which helps to maintain and/or improve
neurological and brain function and/or to prevent and/or improve
cognitive changes and to prevent and/or treat cognitive disorders
like dementia. Preferably, dementia is Alzheimer disease. The
cognitive changes may be stress or age related cognitive changes.
These changes may be mild.
[0010] In particular, the invention relates to vicenin 2 or a
biologically active analogue thereof as an active ingredient for
use in maintaining and/or improving neurological and/or brain
function and/or preventing, delaying onset, controlling and/or
treating a neurological dysfunction, condition, disorder or
disease.
[0011] Preferably, the neurological condition is associated with
impaired neurological and/or brain function. In other words,
preferably, the maintaining and/or improving neurological and/or
brain function may be associated with a neurological dysfunction,
condition, disorder or disease.
[0012] Thus, the present invention may relate to vicenin 2 or a
biologically active analogue thereof as an active ingredient for
use in preventing, delaying onset, controlling and/or treating a
neurological dysfunction, disorder, disease or condition, which is
associated with impaired neurological and/or brain function.
[0013] In a further embodiment, the invention relates to a use of
vicenin 2 or a biologically active analogue thereof as an active
ingredient for maintaining and/or improving neurological and/or
brain function. This embodiment relates to a use of vicenin 2 or a
biologically active analogue thereof for healthy individuals, who
wish to e.g. enhance cognitive performance and/or improve
well-being at stress situations.
[0014] In a further embodiment, the invention relates to vicenin 2
or a biologically analogue thereof as an active ingredient for use
in improving cognitive functions, memory, attention, concentration,
alertness, mental flexibility and/or speed, learning, intelligence,
language, problem solving capacity, consciousness, coping with
psychological stress or tension, motivation, mobility, decision
making capacity, reaction time, anxiety and/or mood swings in a
patient suffering from a neurological dysfunction, disorder or
disease.
[0015] Preferably, the neurological dysfunction, condition,
disorder or disease is an age or stress related neurological
dysfunction, anxiety, a cognitive disorder, depression or dementia,
wherein dementia is preferably Alzheimer disease. Furthermore, the
active ingredient is preferably derived from a plant and the active
ingredient is a plant preparation enriched for the active
ingredient. Preferably, the plant is selected from a group
consisting of a Anethum, Perilla, Urtica, Passiflora, Camelia,
Cayaponia, Colocasia, Cydonia, Desmodium, Hordeum, Origanum,
Ocimum, Jatropha, Parkinsonia, Peperomia, Piheranthos, Centaurea,
Indigo, Bomba, Lychnophera, Asplenium, Chinotto, Citrus, Viola,
Trigonella, Rosemary, Peppermint, Thyme, Basil, Sage, Oregano,
Lavandula, Nipponanthemum, Abrus, Viola, Santalum, Oryza,
Scleropyrum, Tulsi, Centaurea, Indigofera, Bombax, Glinus,
Lychnophora and other species belonging to the Lamiacea, Labiatae,
and Urticaceae, Rosales or Malpighiales or a combination of said
plants.
[0016] The plant preparation may be selected from the group
consisting of a leaf preparation, a fruit preparation, a seed
preparation, a stem preparation, a flower preparation, a bud
preparation, a root preparation or a mixture of different parts of
the plant.
[0017] In one embodiment, the active ingredient is an isolated
vicenin 2 or a biologically active analogue thereof obtained by
isolation or chemical synthesis.
[0018] In a preferred aspect of the present invention, the
neurological and/or brain function is selected from the group
consisting of memory, attention, concentration, alertness, mental
flexibility and/or speed, learning, intelligence, language skills,
problem solving capacity, consciousness, coping with psychological
stress or tension, motivation, mobility, decision making capacity,
reaction time and regulation of emotions. The emotions may be e.g.
anxiety and/or mood swings. These functions may be impaired in
association with a cognitive disorder such as neurodegenerative
disorder or dementia.
[0019] The active ingredient may be comprised in a preferred
embodiment in a composition such as a food product, dietary
supplement or medicament, preferably wherein the medicament 1 to
comprises a pharmaceutically acceptable carrier. In these aspects,
the concentration of the active ingredient is from 0.1 .mu.g to 500
.mu.g, preferably from 2.5 .mu.g to 50 .mu.g, preferably from 5
.mu.g to 15 .mu.g or 12 .mu.g to 30 .mu.g, most preferably about 24
.mu.g.
[0020] Most preferably, the active ingredient is administered at a
dose of 0.1-0.5 .mu.g/kg, 0.17-0.42 .mu.g/kg or 0.07-0.3
.mu.g/kg.
[0021] Furthermore, in a preferred embodiment the active ingredient
may be comprised in globules, pellets, powder formulations,
tablets, capsules, stick formulations, sachet formulations or a
fluid. The fluid may be in a bottle with a dropper.
[0022] Preferably, the composition is substantially free of other
plant flavonoids derived from the same plant or other plant
extracts or free of flavonoids of other plants.
[0023] In another preferred embodiment, the composition comprises
one or more further agents capable of maintaining and/or improving
neurological and/or brain function and/or preventing, delaying
onset, controlling and/or treating a neurological dysfunction,
condition, disorder or disease as defined in any one of the
preceding claims. This agent may be a lipid, a lipid containing
omega-3-fatty acid, a physiologically active fatty acid, an
antioxidant, an anti-inflammatory agent, a bulking agent, an immune
system modulatory agent or a vaccine, an antibody, a metal-protein
interaction attenuation agent, a plant preparation, curcumin,
coenzym Q10, L-carnitine, zinc, epigallocatechingallate, thymol,
p-cymere, vinpocetine, hyperzine A, phosphatidylserine, a vitamin,
alpha liponic acid, a TNF alpha inhibitor, a flavonoid, an
anthocyanidin, a biflavonoid, a flavon, a flavonglycoside or a
carboxylic acid.
[0024] Preferably, the plant preparation is selected from one or
more extracts from a group consisting of an extract of Ginkgo
biloba, Hypericum perforatum, Hyperzia serrata, Galanthus nivalis,
Salvia officinalis, Panex ginseng, Lippia citriodora, Melissa
officinalis, Passiflora incarnate, Passiflora edulis, Bacopa
monniera, Zingiber officinalis, Leucojum aestrum, Concolulus
pluricaulis and Centella asiatica, Emblica officinalis, Coptidis
Rhizoma, Salvia triloba, Piper nigrum, Trigonella foenum-graecum,
Cimicifuga racemosa, Salvia miltiorrhiza, Rhodiola rosea,
Habranthus jamesonii, Phycella herbertiana, Rhodophiala mendocina,
Zephyranthes filifolia, Stephania pierrei, Kaempfera parviflora,
Stephania venosa, Crocus sativus, Salvia species, Bacopa monnieri,
Centella asiatica, Ptychopetalum olacoides, Withania somnifera,
Coptis chinensis, Mangifera indica, Polygala caudata, Polygala
tenuifolia, Halenia elliptica, Evolvilus alsinoides, Celastrus
paniculatus, Clitoria ternatea, Curcuma longa, Acorus calamus,
Terminalia chebula, Lycoris radiata, Magnolia officinalis, Biota
orientalis, Codonopsis pilosula, Evodia rutaecarpa, Polygonum
multiflorum, Aspalathus linearis, Cyclopia species, Adansonia
digitata, Sclerocarya birrea, Actinidia chinensis, Matricaria
recutita or combinations thereof. In a more preferred embodiment,
the plant preparation is a preparation of Melissa officinalis,
Rhodiole rosea, Magnifera indica or a Cyclopie species or any
combination thereof.
[0025] In a preferred embodiment, the agent capable of maintaining
and/or improving neurological and/or brain function and/or
preventing, delaying onset, controlling and/or treating a
neurological dysfunction, condition, disorder or disease is an
acetylcholinesterase inhibitor, a NMDA (N-methyl-D-aspartate)
receptor inhibitor, a non steroidal anti-rheumatic agent, a
neuroleptic agent, a tricyclic antidepressant, an anti-psychotic
agent, a gab junction inhibitor, a non selective monoaminooxidase
inhibitor, an ABCC1 Transporter, an ADAM 10 protein,
methylthioninium chloride, an antibiotic agent, an antiviral agent,
a gamma secretase inhibitor, a beta secretase inhibitor, an
angiotensin receptor antagonist (AT1 antagonist), a cannabinoid,
allopregnanolone or an insulin sensitizer.
[0026] In a further embodiment the present invention relates to a
process of producing a preparation of the active ingredient as
defined above comprising the steps of: [0027] a) selection of the
raw plant material, [0028] b) preparation of the raw plant
material, [0029] c) applying an extraction process using solvent
extraction and/or filtration techniques, preferably followed by
concentration and/or spray drying of the liquid extract into a
powder, [0030] d) determining the concentration of the active
ingredient; and [0031] e) selecting the preparation comprising the
active ingredient at a concentration of at least 0.01%.
[0032] Preferably, the process comprises an extraction process
using aqueous solvent extraction, more preferably water
extraction.
[0033] Preferably, the solvent is water. In a more preferred
embodiment, the extraction is carried out at a temperature of
90.degree. C. to 125.degree. C. and/or the temperature of the
solvent is 55.degree. C. or above, preferably 70.degree. C. or
above.
[0034] Preferably, the plant is selected from one or more plants of
the group consisting of Anethum, Perilla, Urtica, Passiflora,
Camelia, Cayaponia, Colocasia, Cydonia, Desmodium, Hordeum,
Origanum, Ocimum, Jatropha, Parkinsonia, Peperomia, Piheranthos,
Centaurea, Indigo, Bomba, Lychnophera, Asplenium, Chinotto, Citrus,
Viola, Trigonella, Rosemary, Peppermint, Thyme, Basil, Sage,
Oregano, Lavandula, Nipponanthemum, Abrus, Viola, Santalum, Oryza,
Scleropyrum, Tulsi, Centaurea, Indigofera, Bombax, Glinus,
Lychnophora and other species belonging to the Lamiacea, Labiatae,
Urticaceae, Rosales or Malpighiales or combinations thereof.
[0035] In addition, the invention relates to a preparation of the
active ingredient obtainable by the above described process. The
preparation may be used as an active ingredient for use in
maintaining and/or improving neurological and/or brain function
and/or for preventing, delaying onset, controlling and/or treating
a neurological dysfunction, condition, disorder or disease as
defined in any one of the preceding claims.
[0036] Preferably, the preparation comprises the active ingredient
at a concentration of at least, 0.02%, more preferably, at a
concentration of at least 0.1%, 0.15%, 0.2% or most preferably at a
concentration of at least 0.2%, 0.25% or 0.3%.
[0037] In some cases, the preparation may comprise the active
ingredient at a concentration of at least 0.001%, more preferably,
at a concentration of at least 0.002%, 0.005%, 0.008% or preferably
at a concentration of at least 0.010%, 0.015% or 0.02%.
[0038] In some cases, the preparation may comprise the active
ingredient at a concentration of at least 0.2%, more preferably, at
a concentration of at least 0.5%, 1.5%, 3.0% or most preferably at
a concentration of at least 2.0%, 3.0% or 5.0%.
[0039] Detailed descriptions of conventional extraction methods,
such as those employed herein can be found in the literature, for
example in the book "Industrial Scale Natural Products Extraction",
published in 2011 by Wiley-VCH.
[0040] The analytical method to determine the vicenin 2
concentration may be e.g. a chromatographic method called high
performance liquid chromatography. High performance liquid
chromatography (HPLC) is one of the most popular techniques of
analytical chemistry. A UV detector is used, detecting at 320 nm.
For determining the concentration of vicenin 2, a vicenin 2
reference or its derivates may be used as reference material. For
example, apigenin may be used as reference substance and the
vicenin 2 content is calculated via the difference in molecule
weight and the slope and intercept of the calibration curve for
apigenin.
[0041] In the process, the raw plant material is preferably Perilla
species, more preferably Perilla frutescens Britton var. crispa or
var. acuta Kudo. The raw plant material is preferably prepared by
drying, cutting and/or milling. Extraction can be preferably done
with a raw material which particle size is reduced to lower than 2
mm.sup.2. The solvent may be, preferably, water, methanol, ethanol,
propanol, isopropanol, ethyl acetate, hexane, chloroform or
dichloromethane. Most preferably an aqueous solvent, preferably
water, is used since glycosides, like vicenin 2, which have
hygroscopic properties, are extracted by hot water more efficiently
than alcohol. Moreover, some volatile, allergenic compounds like
Perillaldehyde, Methyleugenol and Myristicin, which may occur in
targeted species containing vicenin 2, can be eliminated by hot
water.
[0042] Ratio of raw material to solvent is preferably between 1:100
to 20:100 and more preferably 2:100 to 10:100.
[0043] Extraction can be preferably done by room temperature to up
to 150.degree. C. Extraction is more preferably carried out from
90.degree. C. to 125.degree. C. In a further preferred embodiment,
heat with additional pressure can be used.
[0044] In a preferred embodiment the extraction time is 10 min to 2
hours, more preferably from 20 min to 50 min.
[0045] Preferably, the plant for the process is selected from one
or more plants from the group consisting of Anethum, Perilla,
Urtica, Passiflora, Camelia, Cayaponia, Colocasia, Cydonia,
Desmodium, Hordeum, Origanum, Ocimum, Jatropha, Parkinsonia,
Peperomia, Piheranthos, Centaurea, Indigo, Bomba, Lychnophera,
Asplenium, Chinotto, Citrus, Viola, Trigonella, Rosemary,
Peppermint, Thyme, Basil, Sage, Oregano, Lavandula, Nipponanthemum,
Abrus, Viola, Santalum, Oryza, Scleropyrum, Tulsi, Centaurea,
Indigofera, Bombax, Glinus, Lychnophora and other species belonging
to Lamiaceae, Labiatae, Urticaceae, Rosales or Malpighiales or
combinations thereof.
[0046] The process of the invention provides an extract with a
higher concentration of the active ingredient as the prior art
processes due to concentration of the active ingredient. The higher
concentration is achieved using state of the art extraction
equipment, which allows an optimal interaction between the raw
material and the extraction solvents. All critical process steps,
for example temperature, are controlled in process at any times and
adaptations are continuously possible to ensure highest yield for
the active ingredient. Due to the selection of the extraction
solvents combined with optimized physical conditions it is possible
to diminish unwanted substances which may occur naturally in the
different raw materials.
[0047] In one embodiment, the invention provides a method for
maintaining and/or improving neurological and/or brain function
and/or for preventing, delaying onset, controlling and/or treating
a neurological dysfunction, condition, disorder or disease in a
subject in need thereof comprising administering an active
ingredient as defined above, the composition as defined above or
the preparation as defined above. Preferably, the neurological
dysfunction, condition, disorder or disease is an age or stress
related neurological dysfunction, anxiety, a cognitive disorder,
depression or dementia, wherein preferably the dementia is
Alzheimer disease.
[0048] In a preferred embodiment, the neurological and/or brain
function is selected from the group consisting of memory,
attention, concentration, alertness, mental flexibility and/or
speed, learning, intelligence, language, problem solving capacity,
consciousness, coping with psychological stress or tension,
motivation, mobility, decision making capacity, reaction time and
regulation of emotions.
[0049] In addition, the present invention provides a kit comprising
an active ingredient as defined above, the composition as defined
above or the preparation as defined above and instructions for
administering said composition. Preferably, the kit is for use in
maintaining and/or improving neurological and/or brain function
and/or preventing, delaying onset, controlling and/or treating a
neurological dysfunction, condition, disorder or disease.
[0050] The present invention further provides a composition
comprising the active ingredient as defined above, the preparation
as defined above or the composition as defined above for use in
preventing, delaying onset, controlling and/or treating a disorder
or disease associated with autonomous neuronal death by blocking
the gap junction hemichannel of activated microglia and reducing
their glutamate release. Preferably, said disorder or disease is a
neurodegenerative disorder, such as Alzheimer disease, Parkinson
disease, Huntington disease, multiple sclerosis, amylotrophic
lateral sclerosis (ALS), viral encephalitis or AIDS. Preferably,
said composition comprises or is derived from a plant preparation
comprising the active ingredient. More preferably, the plant
preparation is derived from a plant and/or plant preparation as
defined above. In this embodiment the composition is preferably
Perilla extract.
[0051] In a further embodiment, the present invention relates to a
composition comprising the active ingredient, i.e. vicenin 2 or
biologically active analogue as thereof as defined above or the
preparation as defined above, or the composition as defined above
for use in preventing, delaying onset, controlling and/or treating
a condition associated with glutamate and tumor necrosis factor
.alpha. released by activated microglia inducing excitotoxic
neuronal death. In this embodiment, the composition is preferably
Perilla extract.
[0052] Furthermore, the present invention relates to a composition
comprising the active ingredient having one or more effects,
preferably more than one, more preferably more than two, more than
three, more than four or more than five effects, selected from the
group consisting of reversible acetylcholinesterase inhibition,
blockade of gap junction hemichannels, neuroleptic effect,
anti-depressive effect, enkephalin like effect and reduction in TNF
alpha for use in maintaining and/or improving neurological and/or
brain function and/or preventing, delaying onset, controlling
and/or treating neurological dysfunction, condition, disorder or
disease. Preferably, said condition is associated with impaired
neurological and/or brain function.
[0053] In a further embodiment, the invention relates to a vicenin
2 or a functionally active derivative thereof as an active
ingredient for use in maintaining and/or improving neurological
and/or brain function and/or preventing, delaying onset,
controlling and/or treating a neurological dysfunction, condition,
disorder or disease, wherein the active ingredient is not derived
from Perilla.
BRIEF DESCRIPTION OF THE FIGURES
[0054] FIG. 1: Acute effects of vicenin 2 (AUC-V) on the cortical
network activity in vitro. Plotted are the spike rate changes for
treatment of 9 accumulating concentrations in the range of 10 pg/ml
to 300 .mu.l/ml (mean.+-.standard error, n=17; Student's paired
t-test: * p.ltoreq.0.05: ** p.ltoreq.0.01; *** p.ltoreq.0.001).
[0055] FIG. 2: Acute effects of Perilla leaf extract (AUC-P) on the
cortical network activity in vitro. Plotted are the spike rate
changes for treatment of 9 accumulating concentrations in the range
of 10 ng/ml to 1 mg/ml (mean.+-.standard error, n=5; Student's
paired t-test: * p.ltoreq.0.05; ** p.ltoreq.0.01; ***
p.ltoreq.0.001).
[0056] FIG. 3: A dose-depending effect of a plant extract
comprising vicenin 2 was shown to reduce TNF.alpha.. TNF alpha
secretion [pg/ml] after co-incubation with LPS and Perilla extract
of different concentration (1 .mu.g/ml, 2.5 .mu.g/ml and 50
.mu.g/ml) against 450 .mu.g/ml a benchmark blend, containing e.g.
Boswellia extract, Omega 3 fatty acids, ALA and Curcumin.
DETAILED DESCRIPTION OF THE INVENTION
[0057] The inventors surprisingly found an agent having beneficial
effects on neurological function and brain health, like cognitive
functions, including memory, attention, concentration, alertness,
mental flexibility and/or speed, learning, intelligence, language,
problem solving capacity, consciousness, coping with psychological
stress or tension, motivation, mobility, decision making capacity
and reaction time as well as emotions like anxiety and mood
swings.
[0058] In particular, the inventors surprisingly found that vicenin
2 or functionally active derivatives, i.e. biologically active
analogues, thereof have beneficial effects to maintain and improve
neurological and brain function and to prevent and improve age or
stress related cognitive changes, which may be mild, and to prevent
and treat cognitive disorders, for example dementia, in particular
Alzheimer disease. This efficacy is very beneficial for cognitive
functions, including memory, attention, concentration, alertness,
mental flexibility and/or speed, learning, intelligence, language
skills, problem solving capacity, consciousness, coping with
psychological stress or tension, motivation, mobility, decision
making and reaction time as well as emotions like anxiety and mood
swings.
[0059] Although Alzheimer disease develops differently for every
individual, there are many common symptoms. Early symptoms are
decrease in memory, concentration and alertness, escalating into
confusion, mood swings and emotional difficulties, loss in
orientation, language problems and loss of long-term memory. Even
early symptoms lead to a disconnection to family and society, which
triggers mood swings, including aggression as well as anxiety. In
the advanced status of Alzheimer disease body functions decrease
ultimately leading to death. [4]
[0060] The pathogenesis of Alzheimer disease is not completely
understood and a combination of several causes may lead to the
development of plaques in the brain, which decrease the efficiency
of neuronal communication leading to die off of neurons.
Scientifically these plaques are called "beta amyloid peptides" or
"Tau-proteines". Recent studies confirmed that a genetic
disposition may be involved and/or infection may contribute to the
formation of plaques and the development of Alzheimer disease.
[3,4]
[0061] First-line therapy for all forms of dementia, including
Alzheimer, is intellectual activities, such as reading, playing
board games, completing crossword puzzles, playing musical
instruments, or regular social interaction. Life experiences result
in more efficient neural functioning providing the individual a
cognitive reserve that delays the onset of dementia manifestations.
Physical activity may also be associated with a reduced risk of
dementia and the slowdown in progression. Psychosocial therapies to
reduce daily stress and anxiety are as important as dietary
modifications to increase the intake of antioxidants. [4]
[0062] Several dietary supplement products are available to support
brain health. Most of them are linked to antioxidant and anti-aging
efficacy, for example, long chain omega 3 fatty acids, an
physiological active fatty acid, eicosapenteanoic acid EPA,
docosahexeanoic acid DHA, Ginkgo biloba extract, Curcumin or other
polyphenols, catechins, flavonoids or phenolic carboxylic acid
based plant preparations, including TCMs. Choline and Uridine
monophosphate are also used as well as Vitamins, like folic acid
and the pyridoxine (B6) and B12, vitamin C, vitamin E and selenium.
The functional food products for these areas can also be defined as
medical food which use and intake is to be recommended and
supervised by a medical doctor. The medical food market segment is
a developing market segment.
[0063] The invention provides a new important possibility of
application for brain health, to maintain and improve neurological
and brain function and to prevent and improve age or stress related
associated cognitive changes, which may be even mild, and to
prevent and treat cognitive disorders, dementia and Alzheimer
disease.
[0064] Surprisingly, as shown by the Examples the active ingredient
of the invention and the composition comprising the active
ingredient demonstrate a plurality of modes of action beneficial
for neurological and/or brain function. Thus, the composition
comprising the active ingredient can be used for treating an
individual suffering from several conditions. These may include
conditions associated with reversible acetylcholinesterase
inhibition, blockade of gap junction hemichannels and reduction in
TNF alpha. Furthermore, the active ingredient as well as the
composition comprising the same show neuroleptic effect and
anti-depressive effects. Therefore, the active ingredient and
compositions comprising the same result in a higher success rate in
the treatment of an individual suffering from a neurological
dysfunction, condition, disorder or disease. The disorder may be a
neurodegenerative disorder, preferably Alzheimer disease, Parkinson
disease, Huntington disease, multiple sclerosis, amylotrophic
lateral sclerosis (ALS), encephalitis or AIDS.
[0065] In addition to above, the active ingredient may be used in
treatment of individuals suffering from more than one dysfunctions,
conditions, diseases or disorders. These may include e.g. dementia,
in particular Alzheimer disease, a disorder associated to neuronal
cell death or inflammation.
[0066] A further advantage of the composition of the invention
comprising the active ingredient is that it has a plurality of
therapeutic properties; i.e. the active ingredient or composition
comprising the same has one or more of the following properties (i)
an anti-inflammatory effect, (ii) anti-nociceptic effect, (iii)
sedative effect, (iv) anxiolytic effect, (v) anti-cancer effect,
(vi) immuno-modulation inducing effect and/or (vii) a beneficial
effect on cognitive behavior and/or mood.
DEFINITIONS
[0067] It is to be noted that the term "a" or "an" entity refers to
one or more of that entity; for example, "an antibody", is
understood to represent one or more antibodies. As such, the terms
"a" (or "an"), "one or more", and "at least one" can be used
interchangeably herein.
[0068] By an "isolated" ingredient, variant, or derivative thereof
is intended an agent that is not in its natural milieu. No
particular level of purification is required. For example, an
isolated active ingredient can be removed from its native or
natural environment. Synthetically produced active ingredients are
considered "isolated" for purpose of the invention, as are native
or synthetic active ingredients which have been separated,
fractionated, or partially or substantially purified by any
suitable technique.
[0069] By "subject" or "individual" or "animal" or "patient" or
"mammal", is meant any subject, particularly a mammalian subject,
e.g., a human patient, for whom diagnosis, prognosis, prevention,
or therapy is desired.
[0070] "Maintaining a healthy brain" according to the invention can
be understood as maintaining a normal neurological and brain
function, wherein the normal cognitive function encompasses several
domains, including memory, attention, concentration, alertness,
mental flexibility and/or speed, learning, intelligence, language,
problem solving capacity, consciousness, coping with psychological
stress or tension, motivation, mobility, decision making and
reaction time as well as emotions like anxiety and mood swings.
[0071] "Cognitive changes" relate to changes, for example, in
memory, attention, concentration, alertness, mental flexibility
and/or speed, learning, intelligence, language, problem solving
capacity, consciousness, coping with psychological stress or
tension, motivation, mobility, decision making and reaction time as
well as emotions like anxiety and mood swings. These changes may be
age or stress related. Furthermore, the changes may be mild,
moderate or strong.
[0072] "A neurological condition" may refer to a state of an
individual having one or more impaired neurological and/or
cognitive functions. This state is milder than a disorder or
disease, and may be caused, for example, by age or stress.
[0073] "Prevention of dementia, in particular Alzheimer disease, or
improvement of the status of dementia, in particular Alzheimer
disease" according to the invention can be understood as that the
active ingredient has beneficial physiological effects to prevent
or improve neurological function, cognitive functions, including
memory, attention, concentration, alertness, mental flexibility
and/or speed, learning, intelligence, language, problem solving
capacity, consciousness, coping with psychological stress or
tension, motivation, mobility, decision making and reaction time as
well as emotions like anxiety and mood swings.
[0074] "Delaying onset of a dysfunction, condition, disorder or
disease" can be understood as shifting the beginning of the
dysfunction, condition, disorder or disease to a later time
point.
[0075] "Controlling a dysfunction, condition, disorder or disease"
means keeping or stabilizing a dysfunction, condition, disorder or
disease at a level, which does not worsen dramatically.
[0076] "Dementia" means in this context a significant loss of
intellectual abilities such as memory capacity, severe enough to
interfere with social or occupational functioning.
[0077] "Alzheimer disease" means in this context a progressive
neurologic disease of the brain that leads to the irreversible loss
of neurons and dementia. The clinical hallmarks of Alzheimer's
disease are progressive impairment in memory, judgment, decision
making, orientation to physical surroundings, and language.
[0078] "Neurodegenerative diseases" include i.a. Alzheimer disease,
Parkinson disease, Huntington disease, multiple sclerosis,
amylotrophic lateral sclerosis (ALS), encephalitis or AIDS.
[21]
[0079] Vicenin 2 is a flavonoid having a number of derivatizations.
Vicenin 2 or its synonyms Apigenin-6,8-di-C-glycoside,
5,7,4'-Trihydroxyflavone-6,8-di-C-glucoside,
5,7-Dihydroxy-2-(4-hydroxyphenyl)-6,8-bis[(2S,3R,4R,5S,6R)-3,4,5-trihydro-
xy-6 (hydroxymethyl)oxan-2-yl]chromen-4-one with the CAS Registry
Number 23666-13-9 is a flavonoid found in a number of plant
species. Vicenin 2 can be found i.a. in Anethum, Perilla species,
Urtica species, Passiflora species, Camelia species, Cayaponia
species, Cydonia species, Colocasia species, Desmodium species,
Hordeum species, Origanum species, Ocimum species, Jatropha
species, Parkinsonia species, Peperomia species, Piheranthos
species, Centaurea species, Indigo species, Bomba species,
Lychnophera species, Asplenium species, Chinotto species, Citrus
species, Viola species, Trigonella species, species belonging to
the Lamiacea, Labiatae and e.g. Rosemary, Peppermint, Thyme, Basil,
Sage, Oregano, Lavandula, Nipponanthemum, Abrus, Viola, Santalum,
Oryza, Scleropyrum, Tulsi, Centaurea, Indigofera, Bombax, Glinus,
Lychnophora and species belonging to Urticaceae, Rosales or
Malpighiales.
[0080] The standard process to purify vicenin 2 out of plant
material is based on different chromatographic methods known by
person skilled in the art and consequently, vicenin 2 may be
identified by spectroscopy. These processes can be reviewed in
several publications. [1,3]
[0081] So far the literature describes that vicenin 2 has an
anti-cancer, anti-inflammatory and anti-nociceptive effect. [12,
13, 14]
[0082] "A plant extract comprising vicenin 2 or biologically active
analogue thereof" is herein understood according to this invention
to be an extract comprising vicenin 2 at a concentration which can
be measured. The plant extract or preparation preferably comprises
the active ingredient preferably at a concentration of at least
0.001%, 0.002%, 0.005%, 0.008%, 0.010% or at least 0.015% or from
0.02% to 0.3%, more preferably from 0.1% to 0.2% and preferably
from 0.15% to 0.2%.
[0083] In some cases, the preparation may comprise the active
ingredient at a concentration of at least 0.001%, more preferably,
at a concentration of at least 0.002%, 0.005%, 0.008% or preferably
at a concentration of at least 0.010%, 0.015% or 0.02%.
[0084] In some cases, the preparation may comprise the active
ingredient at a concentration of at least 0.2%, more preferably, at
a concentration of at least 0.5%, 1.5%, 3.0% or most preferably at
a concentration of at least 2.0%, 3.0% or 5.0%.
[0085] As used herein "biologically active derivatives",
"biologically active analogues", "functionally active derivatives"
or "functionally active analogues", which are used interchangeably
herein, of vicenin 2 relate to structurally similar compounds to
vicenin 2, e.g. flavone c-glycosides or flavone o-glycosides, in
particular, apigenin 7-O-.beta.-glucuronide,
apigenin-7-O-[.beta.-glucuronosyl (1.fwdarw.2 .beta.-glucuronide],
luteolin 7-O-[.beta.-glucuronosyl(1.fwdarw.2) .beta.-glucuronide],
luteolin 7-O-.beta.-glucuronide or scutellarein
O-.beta.-glucuronide (scutellarin), which also show effects
beneficial for neurological function and brain health. These
effects may include reversible acetylcholinesterase inhibition,
blockade of gap junction hemichannels and reduction in TNF alpha.
The beneficial effects to neurological function and brain health
can be measured as shown in the examples.
[0086] There are several possibilities to investigate neurological
effects, which are known to the skilled person. In vitro studies,
like receptor binding or enzyme inhibition studies, are standard
methods known to the person skilled in the art to identify activity
and its mode of actions. This kind of test is shown in example 2.
In addition it is possible to apply methods, which are known to the
person skilled in the art and commonly used within the preclinical
development phase of pharmaceutical agents in order to determine
neurological activity when testing the acute neuroactive effects of
substances on neuronal networks of murine cortex being grown on
micro chips. By means of electrophysiological multi-channel
recording, electrical potential are compared to benchmarks to
identify potential effects as well as side effects. This kind of
test is shown in example 1. [15, 16, 17]
[0087] Furthermore, in vivo animal and in vivo human studies are
used to confirm the effect within the biological system. There are
many well established animal tests to investigate neurological
abilities like for example the forced swimming test. [1,8]
Computer-based test for the assessment of working and short-term
memory and selective attention are many times used to assess
cognitive function and progression in cognitive disorders. [1,9]
These kinds of studies are well known for the person skilled in the
art.
[0088] "The active ingredient" as used herein relates to vicenin 2
or functionally or biologically active derivative or analogue
thereof.
[0089] In order to induce the beneficial neurological and brain
health effects, the concentration of the active ingredient in the
composition is 0.1 .mu.g to 500 .mu.g, preferably of 2.5 .mu.g to
50 .mu.g and more preferably of 5 .mu.g to 15 .mu.g or 12 .mu.g to
30 .mu.g, most preferably about 24 .mu.g vicenin 2.
[0090] The composition may be a plant preparation or plant extract,
most preferably a liquid or powder extract obtained by extraction
and comprising vicenin 2.
[0091] The composition according to the present invention may be
comprised in a functional food product, dietary supplement or in a
drug.
[0092] "A functional food product" according to this invention is
understood to be a food, beverage or infant formular product, which
offers, in addition, to nutritional value a health benefit, which
supports and improves health and wellbeing or helps to reduce the
risk to develop a disease.
[0093] "A dietary supplement product" according to this invention
is a food product in form of a pill, tablet, capsule, pellet,
globule, stick formulation, powder formulation, sachet formulation,
powder or liquid form, which are meant to be taken by mouth, and
contain substances like vitamins, minerals, foods, plant
preparations, amino acids and are intended to supplement the usual
intake of these substances via the normal diet.
[0094] "A medicament/drug/medicine" according to this invention is
any substance with the potential to prevent or cure disease or
enhance physical or mental welfare. If not stated otherwise the
term "drug", "medicine", or "medicament" are used interchangeably
herein and shall include but are not limited to all (A) articles,
medicines and preparations for internal or external use, and any
substance or mixture of substances intended to be used for
diagnosis, cure, mitigation, treatment, or prevention of disease of
either man or other animals; and (B) articles, medicines and
preparations (other than food) intended to affect the structure or
any function of the body of man or other animals; and (C) articles
intended for use as a component of any article specified in clause
(A) and (B). The term "drug", "medicine" or "medicament" shall
include the complete formula of the preparation intended for use in
either man or other animals containing one or more "agents",
"ingredients", "compounds", "substances" or "(chemical)
compositions" as and in some other context also other
pharmaceutically inactive excipients as fillers, disintegrants,
lubricants, glidants, binders or ensuring easy transport,
disintegration, disaggregation, dissolution and biological
availability of the "drug", "medicine", or "medicament" at an
intended target location within the body of man or other animals,
e.g., at the skin, in the stomach or the intestine. The terms
"agent", "compound" or "substance" are used interchangeably herein
and shall include, in a more particular context, but are not
limited to all pharmacologically active agents, i.e. agents that
induce a desired biological or pharmacological effect or are
investigated or tested for the capability of inducing such a
possible pharmacological effect by the methods of the present
invention.
[0095] "Pharmaceutically acceptable carrier" may include, but are
not limited to aqueous non-aqueous base solutions, suspensions,
emulsions, microemulsions, micellar solutions, gels and ointments.
The pharmaceutically acceptable carrier may also contain
ingredients that include, but are not limited to, saline and
aqueous electrolyte solutions; ionic and nonionic osmotic agents
such as sodium chloride, potassium chloride, glycerol, and
dextrose; pH adjusters and buffers such as salts of hydroxide,
hydronium, phosphate, citrate, acetate, borate, and tromethamine;
antioxidants such as salts, acids and/or bases of bisulfite,
sulfite, metabisulfite, thiosulfite, ascorbic acid, acetyl
cysteine, cystein, glutathione, butylated hydroxyanisole, butylated
hydroxytoluene, tocopherols, and ascorbyl palmitate; surfactants
such as lecithin, phospholipids, including but not limited to
phosphatidylcholine, phosphatidylethanolamine and phosphatidyl
inositiol; poloxamers and poloxamines, polysorbates such as
polysorbate 80, polysorbate 60, and polysorbate 20, polyethers such
as polyethylene glycols and polypropylene glycols; polyvinyls such
as polyvinyl alcohol and povidone; cellulose derivatives such as
methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose,
carboxymethyl cellulose and hydroxypropyl methylcellulose and their
salts; petroleum derivatives such as mineral oil and white
petrolatum; fats such as lanolin, peanut oil, palm oil, soybean
oil; mono-, di-, and triglycerides; polymers of acrylic acid such
as carboxypolymethylene gel, and polysaccharides such as dextrans,
and glycosaminoglycans such as sodium hyaluronate. Such
pharmaceutically acceptable carriers may be preserved against
bacterial contamination using well-known preservatives, these
include, but are not limited to, benzalkonium chloride, ethylene
diamine tetra-acetic acid and its salts, benzethonium chloride,
chlorhexidine, chlorobutanol, methylparaben, thimerosal, and
phenylethyl alcohol, or may be formulated as a non-preserved
formulation for either single or multiple use.
[0096] As used herein, the terms "treat" or "treatment" refer to
both therapeutic treatment and prophylactic or preventative
measures, wherein the object is to prevent or slow down (lessen) an
undesired physiological change or disorder, such as the development
of a cognitive dysfunction, condition, disorder or disease.
Beneficial or desired clinical results include, but are not limited
to, alleviation of symptoms, diminishment of extent of disease,
stabilization (i.e., not worsening) state of disease, delay or
slowing of disease progression, amelioration or palliation of the
disease state, and remission (whether partial or total), whether
detectable or undetectable. Those in need of treatment include
those already with the condition or disorder as well as those prone
to have the condition or disorder or those in which the
manifestation of the condition or disorder is to be prevented.
[0097] These and other embodiments are disclosed and encompassed by
the description and examples of the present invention. Further
literature concerning any one of the materials, methods, uses and
compounds to be employed in accordance with the present invention
may be retrieved from public libraries and databases, using for
example electronic devices. For example the public database
"Medline" or "Pubmed" may be utilized, which is hosted by the
National Center for Biotechnology Information and/or the National
Library of Medicine at the National Institutes of Health. Further
databases and web addresses, such as the virtual library
"Martindale's center" are known to the person skilled in the art
and can also be obtained using internet search engines.
[0098] Several documents are cited throughout the text of this
specification. The contents of all cited references (including
literature references, issued patents, published patent
applications as cited throughout this application and
manufacturer's specifications, instructions, etc.) are hereby
expressly incorporated by reference; however, there is no admission
that any document cited is indeed prior art as to the present
invention.
[0099] The above disclosure generally describes the present
invention. A more complete understanding can be obtained by
reference to the following specific examples which are provided
herein for purposes of illustration only and are not intended to
limit the scope of the invention.
EXAMPLES
[0100] The examples which follow further illustrate the invention,
but should not be construed to limit the scope of the invention in
any way.
[0101] The practice of the present invention will employ, unless
otherwise indicated, conventional techniques of plant biology,
chemistry, biochemistry, physiology and pharmacology which are
within the skill of the art.
Example 1
Vicenin 2 and Perilla Extract Comprising Vicenin 2 and their
Neurological Efficacy
Methods and Compounds
[0102] Purpose of this example was the evaluation of the acute
neuroactive effects of vicenin 2 (named AUC-V) and of Perilla
extract (named AUC-P) comprising and standardized on Vicenin 2 on
the neuronal activity of murine frontal cortex networks in vitro by
means of electrophysiological multi-channel recordings. In this
system, cells or tissues are grown directly on the chip surface and
communicate via chemical and electrical signals. The MEA-neurochip
allows the non-invasive, long-term, multisite recording of
electrical signal patterns of primary neuronal networks. Said
MEA-neurochip technology enables the characterization of network
activity pattern on the single-cell action-potential level and on
the level of complex neuronal networks as the basic functional
units. This can also be used as a test system for investigating
neuro-physiological properties of compounds. Despite the
complexity, the neurophysiological action profiles of neuroactive
compounds are sensitive and selective as well as robust and stable,
allowing a precise pharmacological "fingerprinting" and the
creation of a database of information on well-characterized
neuroactive substances. The multiparametric description of the
activity pattern changes caused by treatment of a
brain-region-specific neuronal network is a sophisticated approach
to quantify the complex effects of neuroactive agents, of unknown
compounds, and of complex mixtures such as biological extracts.
Correlation with well-known neuroactive substances and their
specific pattern, available in specific databased, provides novel
insights into the possible pharmacological and physiological
mechanisms of modes of action of herbal preparations or isolated
substances. [15, 16, 17]
[0103] In a first step the dose-effect curve of vicenin 2 (AUC-V)
and the plant extract (AUC-P) were plotted by means of cumulatively
increasing the substance concentration, so that the spectrum of
activity of the substance is optimally covered with 9
concentrations. Concentration for Vicenin 2 from 100 fg/ml, 10
pg/ml, 100 pg/ml, 1 ng/ml, 100 ng/ml, 1 .mu.g/ml, 10 .mu.g/ml, 100
.mu.g/ml and 300 .mu.g/ml were used. Concentration for the plant
extract from 10 ng/ml, 100 ng/ml, 1 .mu.g/ml, 10 .mu.g/ml, 30
.mu.g/ml, 100 .mu.g/ml, 200 .mu.g/ml, 500 .mu.g/ml, 1 mg/ml were
used.
[0104] Subsequently, the measurements of the dose-effect curve of
the test substances were repeated at least 10 times. The recorded
electrical activity patterns were characterized by 200 features and
their changes were statistically evaluated.
[0105] Afterwards, a further analysis was carried out through a
pattern-recognition analysis and through comparison with NeuroProof
database to determine relevant mechanisms involved in the frontal
cortex activity pattern induced changes by each substance.
Materials
[0106] The chemicals 5-fluoro-2'-deoxyuridine+uridine (FDU), and
poly-D-lysine were ordered from Sigma-Aldrich Chemical GmbH
(Steinheim, Taufkirchen, Germany). DNase I (from bovine pancreas),
and laminin were purchased from Roche (Mannheim, Germany), fetal
bovine serum from Pan Biotech GmbH (Aidenbach, Germany), and
accutase from PAA (Germany). Horse serum and Dulbecco's Modified
Essential Medium (DMEM) were ordered from GIBCO BRL (Paisley,
UK).
Microelectrode Array Neurochips
[0107] The microelectrode array neurochips (MEA neurochips) were
provided by the Center for Network Neuroscience (CNNS) at the
University of North Texas. These 5.times.5 cm.sup.2 glass chips
have a central recording matrix with 64 passive electrodes and
indium tin oxide conductors. The hydrophobic insulation material
surface was activated by a brief butane flame pulse through a
stainless steel mask. Thus, cell attachment on a confined adhesive
region (5 mm diameter centered on the electrode array) is ensured.
The activated surface regions were coated with poly-D-lysine (25
.mu.g/ml; 30-70 kD) and laminin (16 .mu.g/ml). Fabrication
techniques and culture methods have been described previously.
Cell Culture
[0108] Frontal cortex tissue was harvested from embryonic day 15,
or day 14 chr:NMRI mice. After ethyl ether anesthesia, mice were
sacrificed by cervical dislocation according to the German Animal
Protection Act .sctn.4. Neuronal tissue was cultured including the
use of DNase I (8000 units/ml) and accutase (10 U/ml) for tissue
dissociation. The tissue was dissociated enzymatically with
accutase and mechanically with transfer pipettes. The cells were
resuspended in DMEM 10/10 (10% horse and 10% fetal calf serum) at a
density of 1.0.times.10.sup.6 cells/ml, and 400 .mu.l were seeded
onto MEA surfaces. Cultures were incubated at 37.degree. C. in a
10% CO.sub.2 atmosphere until ready for use, which usually is four
weeks to three months after seeding. Culture media were replenished
three times a week with DMEM containing 10% horse serum. Like in
the tissue of origin, networks develop from a mixture of different
types of postmitotic neurons and glial cells. The glial cells have
important auxiliary functions for the metabolism and for supplying
the neurons with ions and nutrients. The developing co-cultures
were treated with 5-fluoro-2'-deoxyuridine (25 .mu.M) and uridine
(63 .mu.M) for 48 h to prevent further glial proliferation.
[0109] The cells growing directly on the neurochips emerge as
natural neuronal networks. These are composed of a mixture of
neurons and glial cells comparable to the tissue of origin, whereas
in interaction with the neurons, the glial cells fulfill various
metabolism and transport functions. The neurons were coupled
electrically to the neurochip electrodes whereby the action
potentials of the cells can be recorded and their amplitudes and
the electrical activity pattern can be evaluated.
[0110] Activity starts after approximately three to four days in
vitro in form of random spiking. Only after establishing a stable
activity pattern after 4 weeks, the neuronal networks are employed
in substance testing. For this study, cultures between 26 and 29
days in vitro were used.
Multichannel Recording
[0111] For extracellular recording, MEA neurochips were placed into
sterilized constant-bath recording chambers and maintained at
37.degree. C. Recordings were made in DMEM/10% horse serum. The pH
was maintained at 7.4 with a continuous stream of filtered,
humidified airflow with 10% CO.sub.2. Sets of preamplifiers were
positioned to either side of the recording chamber. Recording was
performed with the multichannel acquisition processor system, a
computer-controlled 64-channel amplifier system (Plexon, Inc.,
Dallas, Tex., USA) providing programmable amplification, filtering,
switching, and digital signal processing of microelectrode signals.
The total system gain used was 10K with a simultaneous 40 kHz
sampling rate. The signals routinely recorded by these neurochips
are located in a range of 15-1800 .mu.V.
[0112] The multichannel signal acquisition system delivered single
neuron spike data. Spike identification and separation were
accomplished with a template-matching algorithm in real time. This
allowed the extracellular recording of action potentials from a
maximum of 256 neurons simultaneously.
[0113] The action potentials, or "spikes", were recorded in spike
trains and are clustered in so-called bursts. Bursts were
quantitatively described via direct spike train analysis using the
program NeuroEXplorer (Plexon Inc., Dallas, Tex., USA) and in-house
programs. Bursts were defined by the beginning and end of short
spike invents. Maximum spike intervals defining the start of a
burst were adjusted from 50 to 150 ms and maximum intervals to end
a burst from 100 to 300 ms.
Multiparametric Data Analysis
[0114] The high content analysis of the network activity patterns
provided a multiparametric description characterizing the changes
in four categories: general activity, burst structure,
synchronicity and oscillatory behavior. The substance-specific
activity changes were quantified by calculating for each stable
activity phase after substance application a total of 200
activity-describing spike train parameters for these four
categories below.
Statistical Analysis
[0115] Results are expressed as series means.+-.SEM. The absolute
parameters' distributions were tested for normality. The level of
significance after compound application was assessed using
Student's paired t-test. Significance between two substances or
relative to a contained solvent (e.g. DMSO) was assessed using
Student's unpaired t-test. P<0.05 was considered statistically
significant.
[0116] For direct comparability all parameters were normalized for
each experiment and each experimental treatment with regard to the
corresponding values of the reference activity (native or after
receptor blockade if applicable set to 100%).
[0117] For each experiment, the changing spike rate as a function
of the concentration was fitted to a one-sigmoidal or
multiphasic-sigmoidal dose-response curve given by the
equation:
y = y START + y END - y START 1 + 10 [ log ( EC 90 ) - log ( x ) ]
* nH ##EQU00001##
determining the values of the effective concentration causing 10,
50, and 90% effect (EC.sub.10, EC.sub.50, and EC.sub.90) and of the
slope (Hill coefficient nH; describes the slope of the curve: a
high value corresponds to steep decline which might correspond to
functional neurotoxicity). In case of multiphasic response due to
several mechanisms of action, the right term is repeatedly added
for further phases.
Pattern Recognition and Classification
[0118] To clarify the mode of action of the test substance on the
activity of cortical networks these experiments were further
analyzed using methods of pattern recognition. For each stable
concentration activity phase the 200 spike train parameters were
normalized by the native reference activity. These data records
were computed for the test substances and the reference
substances.
[0119] Using a feature selection algorithm, on the basis of the
reference substances, the 40 most descriptive parameters for all
200 spike train parameters were selected. The rankings of activity
features using various score methods based on classification
experiments and comparing their total correct predictions were
calculated. In this manner, the best results for a MDL (minimal
description length) modified algorithm were obtained. A training
data set with these 40 spike train parameters was established in
the form of data records from the reference substances. An
artificial neuronal network, multi layer feed forward network and
back propagation algorithm without hidden units was then trained.
The respective data records of the four substances were all
subsequently classified. A classification against 105 substances in
the database was carried out.
Results:
[0120] Vicenin 2 showed an effect to induce a decrease of the
cortical network activity with an EC.sub.50 at 2 pg/ml and a
maximum decrease to 80% of the native activity with no affects on
the burst structure, however on the oscillatory behaviour of the
network. This means that the undesired overload of the cortical
networks can be diminished by vicenin 2. These results are shown in
FIG. 1, which illustrates the concentration depending changes in
the activity of the cortical network. It could be seen that vicenin
2 has an influence on the neuroactivity.
[0121] In addition, the extract comprising vicenin 2 has an effect
to induce a decrease of the cortical network activity with an
EC.sub.50 at 90 ng/ml and 200 .mu.g/ml and a maximum decrease to
16% of the native activity with no affects on the burst structure,
however on the oscillatory behaviour of the network. These results
are shown in FIG. 2, which illustrates the concentration depending
changes in the activity of the cortical network. It could be seen
that the extract comprising vicenin 2 has an influence on the
neuroactivity.
[0122] In a second step, these activities were further evaluated by
a classification against positive controls.
Classifications for Vicenin 2 and the Perilla Extract
[0123] The classification was carried out against 105 substances
using the electrophysiological multi-channel recording of vicenin 2
as well as the Perilla extract.
TABLE-US-00001 TABLE 1 Classification of vicenin 2 (AUC-V) and the
extract (AUC-P) against the NeuroProof database. Shown is a ranking
of classification results, which means that x-% of all data records
of this substance were classified as the respective substance in
the left column; DS corresponds to the relative overall ranking.
AUC-P, all 122 AUC-V, all 143 reference # reference # Muscimol 25
Olanzapine 17 Eserine 24 Eserine 15 DPDPE 19 Amisulpride 15
Acetaminophen 18 Enkephalin 14 Cortisol 14 Sodium dodecyl sulfate
12 Amitriptyline 13 Atropinemethylbromide 12 Amisulpride 12
Nicotine 12 Sodium propionate 12 Indatraline 12 Quetiapine 11
Modafinil 12 Carbenoxolone 11 DPDPE 11
[0124] For example, it was possible to identify a sedative and
anxiolytic activity of vicenin 2 as well as of the Perilla extract
by the classification against Amisulpride, which is a known
neuroleptica. It was also possible to identify a sedative and
anxiolytic activity for Perilla extract by classification against
Muscimol, which is a known GABA receptor agonist.
[0125] Surprisingly, it was also possible to identify an
enkephalin-like activity for vicenin 2. Enkephalins are endogenous
ligands that bind to the body's opioid receptors to modulate
neuronal function, like learning or emotional behaviour, which can
be altered by neurodegenerative disorders. In Alzheimer disease,
elevated enkephalin levels may reflect compensatory or contribute
to cognitive impairments.
[0126] Surprisingly, vicenin 2 a well as the extract comprising
vicenin 2 showed also an electrophysiological multi-channel
recording comparable with the pattern of the positive control
eserine and nicotine. The extract shows also an
electrophysiological multi-channel recording comparable with the
pattern of the positive control eserine. Eserine and analoga are
used in the treatment of Alzheimer disease as reversible
acetylcholinesterase inhibitors to improve short term memory.
[6,7]
[0127] In addition, surprisingly vicenin 2 and the plant extract
comprising the same demonstrated an electrophysiological
multi-channel recording comparable with the pattern of the positive
control of carboxolone. Carbenoxolone blocks gap junction
hemichannel of activated microglia and reduces their glutamate
release and consequently diminishes cell autonomous neuronal death
in neurodegenerative diseases. Therefore, gap junction inhibitors
are beneficial in treatment of neurodegenerative diseases.
Carboxolone and analoga are used in the treatment of Alzheimer
disease. [10]
[0128] Further, both the standardized extract as well as the
isolated vicenin 2 demonstrated effects similar to amisulpride,
which is a known neuroleptic agent. In addition, the extract showed
an effect to anti-depressive amitripthyline and the active
ingredient vicenin 2 to the anti-depressives and antipsychotics
olanzapine and indatraline. Neuroleptika and antipsychotics are
also used as companying therapy of Alzheimer to increase mood and
motivation and to decrease anxiety and depression. This
demonstrates further the effect of vicenin 2 and a composition
comprising it in neurological and/or brain function and/or in
maintaining and/or improving neurological and/or brain function
and/or preventing, delaying onset, controlling and/or treating a
neurological dysfunction, condition, disorder or disease.
[0129] Thus, example 1 surprisingly shows for the first time that
vicenin 2 and an extract comprising the same (Perilla extract) can
be used as a beneficial agent for neurological and brain functions
or for the treatment of a cognitive disorder. Cognitive functions
include, memory, attention (concentration), alertness, learning,
intelligence, language, problem solving capacity, coping with
psychological stress or tension, anxiety and mood alterations.
Example 2
Perilla Extract Comprising and Standardized on Vicenin 2 has the
Ability to Reduce TNF.alpha.
[0130] This study investigated the efficacy of Perilla extract
comprising vicenin 2 to contribute to reduce TNF.alpha. levels. The
study was designed as an ex vivo study, where human whole blood
samples were treated with LPS, a bacterial blend, to stimulate
inflammation which leads to an increase of the cytokine TNF.alpha..
Furthermore, the effect to reduce TNF.alpha. by Perilla extract was
measured in comparison to a negative and positive control. Perilla
extract demonstrated a dose-depending effect to reduce TNF.alpha.
(see FIG. 3).
[0131] Surprisingly, the extract as well as the isolated vicenin 2
demonstrated anti-inflammatory effects. Perilla extract
demonstrated TNF.alpha. inhibiting efficacy within this ex vivo
study showing a further beneficial effect in treatment if a
cognitive disorder. In addition, TNF.alpha. has been recognized to
be a gliatransmitter that regulates synaptic function in neural
networks. Anti-TNF .alpha. treatment may thus lead to cognitive
improvement. [8]
[0132] Even though anti-inflammatory agents have already been
connected with Alzheimer disease, classical anti-inflammatory
drugs, like NSAR, which act via the prostaglandin pathway most
probably do not work in Alzheimer disease. Inflammation in the
brain is mediated rather by activated microglial cells having an
increased metabolic activity resulting in the production of
proteins, which may contribute to the invasion of plaques. In
addition, they produce inflammatory markers, like cytokines.
Therefore, inhibition of cytokines, like tumor necrose factor-alpha
(TNF.alpha.), antioxidants and anti-inflammatory agents may also
prevent the formation of amyloid beta, thus having a beneficial
effect in Alzheimer disease [8].
[0133] Alzheimer is a multiplex disease and each person reacts
differently on possible treatments. It is difficult to select the
right treatment and therefore, ingredients which combine several
mode of actions are preferred. However, before the present
invention such agents having more than one effective mode of
actions were lacking. Surprisingly, the present inventors showed
that vicenin 2 and functionally active derivatives thereof as well
as plant extracts comprising the same have more than one beneficial
mode of actions for neurological and brain functions. This
inventive agent may combine synergistically several mode of
actions, i.e. at least blocking activity of gap junction
hemichannel of activated microglia, inhibiting activity of tumor
necrose factor .alpha. and inhibiting activity of
acetylcholinesterase as well as having neuroleptic, anti-depressive
effects and enkephaline like effects, and provides a new treatment
possibility to maintain and improve neurological and/or brain
function and to prevent and/or improve cognitive changes and to
prevent and/or treat cognitive disorders, dementia including
Alzheimer disease. The cognitive changes may be age or stress
related and even mild.
[0134] Isolated vicenin 2 and derivates thereof as well as plant
extracts comprising the same may combine synergistically several
mode of actions being beneficial for maintenance and improvement of
neurological and brain functions and for preventing and treating a
cognitive disorder, dementia such as Alzheimer disease. The
combination to offer several mode of actions within one agent is
surprising and extremely beneficial as it contributes to a higher
rate of success in the treatment of such a multi-complex and
inter-individually disease like Alzheimer disease. The invention
offers a highly effective beneficial agent and composition with a
high compliance as individuals prefer to take one agent at once
instead of many.
SUMMARY
[0135] Summarizing, vicenin 2 and biologically active derivatives
thereof as well as composition comprising the same demonstrated
surprisingly beneficial neurological efficacy.
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* * * * *
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