Note: Descriptions are shown in the official language in which they were submitted.
CA 02612595 2007-12-21
Compositions and methods for inducing the expression of heat shock
proteins
Field of the Invention
The present invention relates to a nutritional composition and method for
enhancing heat shock protein expression in cells. Specifically, the present
invention relates to a composition and method comprising a synergistic
combination of at least one substance capable of activating heat shock
transcription factors and Schisandrin B, which act substantially
simultaneously
via differing mechanisms to increase the expression of heat shock proteins in
cells, particularly heat shock protein 72 in skeletal muscle, to facilitate
increased
hypertrophy as a result of exercise.
Background of the Invention
When a mammalian cell is exposed to a sudden elevation in temperature
the expression of most cellular proteins is decreased. However, some proteins,
specifically heat shock proteins (HSP), show increased levels of expression
when cells are subjected to elevated temperatures and other metabolic
stresses.
Examples of metabolic stresses which elicit elevated expression of heat shock
proteins include: decreased glucose availability; increased intercellular
calcium
levels; and decreased blood flow.
Heat shock proteins function as molecular chaperones to prevent protein
aggregation and facilitate the folding of non-native proteins, particularly
new
peptides emerging from ribosomes. Molecular chaperones recognize non-native
proteins, predominantly via exposed hydrophobic residues, and bind selectively
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to those proteins to form relatively stable complexes. In these complexes, the
protein is protected and able to fold into its native form.
Among the many families of heat shock proteins, HSP72, the stress-
inducible protein of the HSP70 family, is one of the best known endogenous
35 factors protecting cells against tissue injury. Research of exercise-
induced
stress response has shown that exercise results in increased expression of
HSP72 mRNA and subsequently in HSP72 protein.
Repetitive, forceful muscular contractions, i.e. physical exercise, cause
changes in the expression patterns of genes and proteins. These changes can
40 result in muscle adaptations such as muscle atrophy via muscle protein
catabolism or muscle hypertrophy via muscle protein accretion. During
hypertrophy, numerous nascent proteins are formed. An increase in the
presence of molecular chaperones, such as HSP72, will act to enhance the
stability of these nascent proteins until they can fold into their native
forms.
45 In situations of enhanced protein turnover, such as the environment in
muscle following exercise, it would be advantageous for an individual to have
a
means of increasing the stability of rapidly forming proteins in order to
reduce the
catabolism of these new non-native state proteins.
Summary of the Invention
50 The present invention relates to a nutritional composition and method for
enhancing heat shock protein expression in cells. The nutritional composition,
comprising an effective amount of at least one substance capable of activating
heat shock transcription factors and an effective amount of Schisandrin B
acting
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synergistically, via differing mechanisms, to increase expression of heat
shock
55 proteins in cells, particularly heat shock protein 72 in skeletal muscle.
Both a
composition and a method are provided by the present disclosure.
Detailed Description of the Invention
In the following description, for the purposes of explanations, numerous
specific details are set forth in order to provide a thorough understanding of
the
60 present invention. It will be apparent, however, to one of ordinary skill
in the art
that the present invention may be practiced without these specific details.
The present invention is directed towards a nutritional composition and
method for enhancing heat shock protein expression in cells. The nutritional
composition, comprising an effective amount of geranylgeranylacetone,
65 paeoniflorin, or a combination thereof, and an effective amount of
Schisandrin B
functioning synergistically, via differing mechanisms, to increase the
expression
of heat shock proteins in cells, particularly heat shock protein 72 in
skeletal
muscle, to facilitate increased hypertrophy as a result of exercise.
A used herein, the term 'nutritional composition' includes dietary
70 supplements, diet supplements, nutritional supplements, supplemental
compositions and supplemental dietary compositions or those similarly
envisioned and termed compositions not belonging to the conventional
definition
of pharmaceutical interventions as is known in the art. Furthermore,
'nutritional
compositions', as disclosed herein, belong to a category of compositions
having
75 at least one physiological function when administered to a mammal by
conventional routes of administration.
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Alternatively, formulations and nutritional compositions belonging to the
present invention may be considered to be nutraceuticals. As used herein, the
term 'nutraceutical' is recognized and used in the art to describe a specific
80 chemical compound or combination of compounds found in, organic matter for
example, which may prevent, ameliorate or otherwise confer benefits against an
undesirable condition. As is known in the art, the term 'nutraceutical' is
used to
refer to any substance that is a food, a part of food, or an extract of food
which is
suitable for consumption by an individual and provides a physiological benefit
85 which may be medical or health-related. Furthermore, the term has been used
to
refer to a product isolated, extracted or purified from foods or naturally-
derived
material suitable for consumption by an individual and usually sold in
medicinal
forms, such as capiets, tablet, capsules, softgel capsules, gelcaps and the
like,
not associated with food.
90 Extracts suitable for use in the present invention may be produced by
extraction methods as are known and accepted in the art such as alcoholic
extraction, aqueous extractions, carbon dioxide extractions, for example.
As used herein, the term 'heat shock protein' is understood to encompass
both proteins that are expressly labeled as such as well as other stress
proteins,
95 including homologs of such proteins that are expressed in the absence of
stressful conditions. Furthermore, as used herein, the term 'heat shock
protein'
is understood to encompass the mRNA species corresponding to expressly
labeled heat shock proteins as well as other stress proteins, which are known
to
be translated into proteins.
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100 Geranylgeranylacetone (GGA)
Geranylgeranylacetone is an acyclic polyisoprenoid that has been used to
protect gastric mucosa. GGA has been shown to activate transcription factors,
particularly heat shock transcription factor (HSF)-1, which are able to bind
to
DNA and induce transcription. HSF-1 is normally suppressed since it is
typically
105 bound to the C-domain of constitutively active HSP70. GGA is able to bind
to the
C-domain of the HSP70 thereby causing HSF-1 to dissociate. HSF-1 is now able
to undergo trimerization and be translocated to the nucleus, where it binds to
the
heat shock-responsive element (HSE) in the promoter region of inducible HSP70
(i.e. HSP72) genes.
110 Recent experiments using cultured mouse skeletal cells, showed that
treatment with GGA up-regulated the expression of HSP 72, and increased
muscular protein content in a dose-dependent manner. Additionally GGA was
shown to facilitate the differentiation of myoblasts into myotubules.
Non-differentiated myoblasts, often referred to as satellite cells, are a
115 small population of quiescent muscle precursor cells that occupy a
"satellite"
position immediately outside of muscle fibers. They are normally maintained in
a
quiescent state and become activated to fulfill roles of routine maintenance,
repair and hypertrophy. Satellite cells are thought to be muscle-specific stem
cells which are capable of producing large numbers of differentiated progeny
as
120 well as being capable of self-renewal. Such that satellite cells can
fulfill their
biological role, they must become activated, proliferate, differentiate and
fuse to
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existing muscle cells. In this way, multinucleate muscle fibers are maintained
or
increased in size in response to stimuli.
It is herein understood by the inventors that inclusion of
125 geranylgeranylacetone or derivatives of geranylgeranylacetone in a
nutritional
composition, will act to increase the expression of heat shock proteins,
particularly HSP72, via directly activating HSF-1. Enhanced expression of heat
shock proteins, particularly HSP72, will act to increase protein accretion via
increased stabilization of nascent proteins. The increased expression of
130 chaperone proteins, i.e. HSP72, in working muscle is important in order to
stabilize the large number of new proteins being synthesized by working
muscle,
leading to increased accumulation of contractile protein, i.e. muscle
hypertrophy.
Additionally, it is herein understood by the inventors that administration of
GGA will have the added benefit of facilitating the differentiation of
myoblasts to
135 myofibers. These myofibers fuse with existing muscle cells thereby
increasing
the size of the muscle cells and ultimately muscle tissue.
As used herein, a serving of the present nutritional composition comprises
from about 1 mg to about 300 mg of geranylgeranylacetone or derivatives of
geranylgeranylacetone. More preferably, a serving of the present nutritional
140 composition comprises from about 25 mg to about 150 mg of
geranylgeranylacetone or derivatives of geranylgeranylacetone. A serving of
the
present nutritional composition most preferably comprises from about 25 mg to
about 75 mg of geranylgeranylacetone or derivatives of geranylgeranylacetone.
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145 Paeoniflorin
Paeoniflorin is a major constituent of peony plants, such as Paeonia
lactoflora, P. suffruticosa, P. obovata, and P. veitchii. The roots of peony
plants
have commonly been used in Chinese medicine to reduce fever and pain, stop
bleeding, prevent infection, and as an antispasmodic.
150 In vitro studies showed that cells treated with paeoniflorin have enhanced
levels of expression of heat shock proteins. Paeoniflorin treatment resulted
in
phosphorylation of HSF-1 allowing HSF-1 to translocate to the nucleus. Inside
the nucleus phosphorylated HSF-1 proteins combine to form granules (trimers)
which have the ability to bind to the HSE region of inducible heat shock
protein
155 genes, thereby inducing transcription of these genes.
It is herein understood by the inventors that inclusion of paeoniflorin or
derivatives of paeoniflorin in a nutritional composition, will act to increase
the
expression of heat shock proteins, particularly HSP72, via directly activating
HSF-1. Paeoniflorin or derivatives of paeoniflorin will enhance the expression
of
160 HSP by increasing the phosphorylation and DNA-binding ability of HSF-1
thereby
facilitating the induction of heat shock proteins. Enhanced expression of heat
shock proteins, particularly HSP72, will act to increase protein accretion via
increased stabilization of nascent proteins. The increased expression of
chaperone proteins, i.e. HSP72, in working muscle is important in order to
165 stabilize the large number of new proteins being synthesized by working
muscle,
leading to increased accumulation of contractile protein, i.e. muscle
hypertrophy.
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As used herein, a serving of the present nutritional composition comprises
from about 1 mg to about 300 mg of paeoniflorin or derivatives of
paeoniflorin.
More preferably, a serving of the present nutritional composition comprises
from
170 about 1 mg to about 150 mg of paeoniflorin or derivatives of paeoniflorin.
A
serving of the present nutritional composition most preferably comprises from
about 1 mg to about 75 mg of paeoniflorin or derivatives of paeoniflorin.
Schisandrin B
Schisandrin B (Sch B) is a dibenzocyclooctadiene compound that is
175 isolated from Schisandrae chinensis. Sch B has been used to enhance the
detoxification of xenobiotics in the liver and assist in liver regeneration.
Recent
studies have shown that Sch B can protect various organs from free-radical
induced damage.
In a study using mice, administration of Sch B was shown to increase the
180 production of HSP70, i.e. HSP72. Treatment with Sch B produces oxidants
via
cytochrome p-450 metabolism, which act as mild stressors to induce HSP70
(HSP72) production.
It is herein understood by the inventors that inclusion of Schisandrin B in a
nutritional composition, will act to increase the production of HSP72, by
185 increasing the production of oxidants from cytochrome P-450 metabolism.
Enhanced expression of HSP72, will act to increase protein accretion via
increased stabilization of nascent proteins. The increased expression of
chaperone proteins, i.e. HSP72, in working muscle is important in order to
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stabilize the large number of new proteins being synthesized by working
muscle,
190 leading to increased accumulation of contractile protein, i.e. muscle
hypertrophy.
As used herein, a serving of the present nutritional composition comprises
from about 1 mg to about 150 mg of Schisandrin B. More preferably, a serving
of
the present nutritional composition comprises from about 1 mg to about 75 mg
of
Schisandrin B. A serving of the present nutritional composition most
preferably
195 comprises from about 1 mg to about 25 mg of Schisandrin B.
In embodiments of the present invention, which are set forth in detail in the
Examples below, the nutritional composition of the present invention comprises
geranylgeranylacetone, paeoniflorin, or a combination thereof, and Schisandrin
B. The nutritional composition is provided in any acceptable and suitable oral
200 dosage form as known in the art. Increased expression of heat shock
proteins is
induced and carried out in an individual by administration of the composition
of
the present invention.
The nutritional composition of the present invention may be administered
in a dosage form having controlled release characteristics, e.g. time-release.
205 Furthermore, the controlled release may be in forms such as a delayed
release
of active constituents, gradual release of active constituents, or prolonged
release of active constituents. Such active constituents release strategies
extend
the period of bioavailability or target a specific time window for optimal
bioavailability. Advantageously the nutritional composition may be
administered
210 in the form of a multi-compartment capsule which combines both immediate
release and time-release characteristics. Individual components of the
nutritional
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composition may be contained in differential compartments of such a capsule
such that the specific components may be released rapidly while others are
time-
dependently released. Alternatively, a uniform mixture of the various
215 components of the present invention may be divided into both immediate
release
and time-release compartments to provide a multi-phasic release profile.
According to various embodiments of the present invention, the nutritional
supplement may be consumed in any form. For instance, the dosage form of the
nutritional supplement may be provided as, e.g., a powder beverage mix, a
liquid
220 beverage, a ready-to-eat bar or drink product, a capsule, a liquid
capsule, a
softgel capsule, a tablet, a caplet, or as a dietary gel. The preferred dosage
form
of the present invention is as a softgel capsule.
Furthermore, the dosage form of the nutritional supplement may be
provided in accordance with customary processing techniques for herbal and
225 nutritional supplements in any of the forms mentioned above. Additionally,
the
nutritional supplement set forth in the example embodiment herein may contain
any appropriate number and type of excipients, as is well known in the art.
The present nutritional composition or those similarly envisioned by one of
skill in the art, may be utilized in methods to enhance the expression of heat
230 shock proteins in cells, particularly heat shock protein 72 in skeletal
muscle,
thereby increasing hypertrophy as a result of exercise.
Although the following examples illustrate the practice of the present
invention in various embodiments, the examples should not be construed as
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limiting the scope of the invention. Other embodiments will be apparent to one
of
235 skill in the art from consideration of the specifications and examples.
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Examples
Example 1:
A nutritional composition comprising the following ingredients per serving are
prepared for consumption as three Softgel Capsules, to be taken twice daily:
240
from about 1 mg to about 300 mg of geranylgeranylacetone, and from about 1
mg to about 150 mg of Schisandrin B.
Example 2:
A nutritional composition comprising the following ingredients per serving are
245 prepared for consumption as four Softgel Capsules, to be taken twice
daily:
from about 1 mg to about 300 mg of geranylgeranylacetone, from about 1 mg to
about 300 mg of paeoniflorin, and from about 1 mg to about 150 mg of
Schisandrin B.
250 Example 3:
A nutritional composition comprising the following ingredients per serving are
prepared for consumption as four Softgel Capsules, to be taken twice daily:
about 50 mg of geranylgeranylacetone, and about 5 mg of Schisandrin B.
255 Example 4:
A nutritional composition comprising the following ingredients per serving are
prepared for consumption as four Softgel Capsules, to be taken twice daily:
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about 50 mg of geranylgeranylacetone, about 10 mg of Schisandrin B, and about
260 100 mg of Ethyl pyruvate.
Example 5:
A nutritional composition comprising the following ingredients per serving are
prepared for consumption as four Softgel Capsules, to be taken twice daily:
265 about 100 mg of geranylgeranylacetone, about 25 mg of Schisandrin B, about
100 mg of Ethyl pyruvate, and about 1 mg of Sulbutiamine.
Example 6:
A nutritional composition comprising the following ingredients per serving are
prepared for consumption as three Softgel Capsules, to be taken twice daily:
270
about 10 mg of geranylgeranylacetone, about 1 mg of Schisandrin B, and about
75 mg of Ethyl pyruvate.
Example 7:
A nutritional composition comprising the following ingredients per serving are
275 prepared for consumption as three Softgel Capsules, to be taken twice
daily:
about 50 mg of geranylgeranylacetone, about 1 mg of paeoniflorin, and about 1
mg of Schisandrin B.
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280 Extensions and Alternatives
In the foregoing specification, the invention has been described with a
specific embodiment thereof; however, it will be evident that various
modifications and changes may be made thereto without departing from the
broader spirit and scope of the invention.
285
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