Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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Composition and Method for Increasing the Anabolic State of Muscle Cells
Field of the Invention
The present invention relates to a nutritional supplement for inducing an
anabolically-favored state for muscle by substantially simultaneously
maintaining
blood and muscle physiological pH levels as well as increasing cellular
concentrations of branched-chain amino acids. More specifically, the present
invention relates to a nutritional supplement comprising a combination of
ethyl
pyruvate and a-hydroxyisocaproic acid (HICA).
Background of the Invention
During intense periods of exercise, where the rate of demand for energy is
high, pyruvate resulting from the breakdown of glucose is converted into
lactate.
This reduction of pyruvate to lactate is beneficial since it regenerates NAD+
for
the continuation of glycolytic energy production required by the working
muscle.
Increased lactate can be removed in a number of ways; it can be exported from
the oxygen-deficient cell and taken up by an oxygen-rich cell where it can be
oxidized to pyruvate and used directly to fuel the citric acid cycle (Brooks
GA.
Mammalian fuel utilization during sustained exercise. Comp Biochem Physiol B
Biochem Mol Biol. 1998 May;120(1):89-107. Review), or it can be reconverted by
the liver, through the Cori cycle, to glucose.
The recognition of monocarboxylate transport (MCT) proteins in the
mitochondria and the closely associated lactate oxidation complexes (Kirkwood
SP, Munn EA, Brooks GA. Mitochondrial reticulum in limb skeletal muscle. Am J
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Physiol. 1986 Sep;251(3 Pt 1):C395-402), suggests that lactate can be
transported and oxidized in the mitochondria of the same cell.
Contrary to popular belief, increased levels of lactate do not directly cause
acidosis; an elevated presence of acidic species (Robergs R, Ghiasvand F,
Parker D. Biochemistry of exercise-induced metabolic acidosis. Am J Physiol
Regul Integr Comp Physiol. 2004;287:R502-16). Lactate appears to be a
consequence rather then the cause of cellular events which cause acidosis. The
physiological state of muscle cells are such that lactate never has hydrogen
available to decrease pH in the surrounding solution. Acidosis is actually a
result
of the hydrolysis of ATP, wherein hydrogen ions are released into the
surrounding solution. During heavy exercise, ATP is produced and utilized
quickly in the cytoplasm causing a rapid decrease in cellular pH. The
buffering
systems of the tissues are rapidly overcome and pH drops resulting in a state
of
acidosis.
Additionally, several hours after exercise there are dynamic changes in
the rates of both skeletal muscle synthesis and breakdown. The consumption of
specific dietary components are known to further influence the response of
skeletal muscle to exercise. The main components of food which are known to
stimulate increased muscle protein synthesis are amino acids (Rennie MJ. Body
maintenance and repair: how food and exercise keep the musculoskeletal
system in good shape. Exp Physiol. 2005 Jul;90(4):427-36). Increased levels of
circulating essential amino acids have been shown to stimulate protein
synthesis
(Smith K, Reynolds N, Downie S, Patel A, Rennie MJ. Effects of flooding amino
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acids on incorporation of labeled amino acids into human muscle protein. Am J
Physiol. 1998 Jul;275(1 Pt 1):E73-8).
More specifically, the branched-chain amino acids (BCAA) consisting of
Leucine, Isoleucine and Valine, are not only used for the synthesis of other
amino acids, but are also important in the regulation of anabolic processes in
muscle. Furthermore, BCAA not only increase the rate of protein synthesis but
also inhibit the rate of protein degradation (Matthews DE. Observations of
branched-chain amino acid administration in humans. J Nutr. 2005 Jun;135(6
Suppl):1580S-4S).
In situations following extended periods of repetitive, forceful muscular
contractions, such as during exhaustive physical exercise, it would be
advantageous for an individual to both maintain physiological pH levels and
increase cellular concentrations of Leucine. In this regard, the anabolic
state of
muscle is increased, facilitating shorter recovery periods as well as
increasing
strength and muscle size.
Summary of the Invention
The present invention is directed towards a nutritional supplement,
comprising at least a therapeutically effective amount of ethyl pyruvate and a
therapeutically effective amount of a-hydroxyisocaproic acid (HICA). The
ingredients of the present nutritional supplement act to induce an
anabolically-
favored state in muscle by substantially simultaneously maintaining blood and
muscle physiological pH levels as well as increasing cellular concentrations
of
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Leucine. Both a composition and a method are provided by the present
disclosure.
In additional aspects of the present invention the a-hydroxyisocaproic acid
(HICA), may be replaced by other a-hydroxy branched-chain amino acid
metabolites, such as a-hydroxy ,B-methylvaleric acid (HIMVA), and a-hydroxy-
isovaleric acid (HIVA), for example. Additionally, the composition of the
present
invention may include one or more of the a-hydroxyisocaproic acid (HICA), a-
hydroxy,(-methylvaleric acid (HIMVA), and a-hydroxy-isovaleric acid (HIVA).
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
present invention. It will be apparent, however, to one of ordinary skill ih
the art
that the present invention may be practiced without these specific details.
The present invention is directed towards a nutritional supplement, for
inducing an anabolically-favored state in muscle by substantially
simultaneously
maintaining blood and muscle physiological pH levels as well as increasing
cellular concentrations of Leucine.
As used herein, the term 'anabolic' is understood to represent metabolic
processes where complex molecules are synthesized from more simple ones, i.e.
synthesis of muscle proteins from amino acids. Additionally, as used herein,
the
term `anabolic' also includes mechanisms of action which are anti-catabolic,
such
as destructive processes wherein the break down or larger molecules into
smaller molecules occurs.
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As used herein, 'a-hydroxyisocaproic acid' makes reference to the
chemical 2-hydroxy-4-methylvaleric acid, (CAS Registry No. 498-36-2), also
known as HICA, or leucic acid. Additionally, as used herein, the term 'a-
hydroxyisocaproic acid' also includes derivatives of a-hydroxyisocaproic acid
such as esters, and amides, and salts, as well as other derivatives, including
derivatives having substantially similar pharmacoproperties to a-
hydroxyisocaproic acid upon metabolism to an active form.
As used herein, the term 'a-hydroxy branched-chain amino acid
metabolite' includes nitrogen-free metabolites of the branched-chain amino
acids,
Leucine, Isoleucine and Valine. More specifically, the term 'a-hydroxy
branched-
chain amino acid metabolite' refers to a-hydroxyisocaproic acid (HICA), a-
hydroxy,a-methylvaleric acid (HIMVA), and a-hydroxy-isovaleric acid (HIVA).
A used herein, the term 'nutritional supplement' includes dietary
supplements, diet supplements, nutritional compositions, supplemental dietary
and other compositions similarly envisioned and termed not belonging to the
conventional definition of pharmaceutical interventions as is known in the
art.
Furthermore, 'nutritional supplements' as disclosed herein belong to category
of
compositions having at least one physiological function when administered to a
mammal by conventional routes of administration.
Ethyl Pyruvate
Pyruvate, or pyruvic acid, is a simple a-ketocarboxylate which is an
important intermediate of glucose metabolism as well as being an endogenous
antioxidant and free radical scavenger (Brand KA, Hermfisse U. Aerobic
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glycolysis by proliferating cells: a protective strategy against reactive
oxygen
species. FASEB J. 1997 Apr;11(5):388-95). This recognition of pyruvate as an
effective free radical scavenger prompted a surge of investigation for
therapeutic
uses.
However, a limitation with regard to the usefulness of pyruvate is its poor
stability in aqueous solution (Sappington PL, Han X, Yang R, Delude RL, Fink
MP. Ethyl pyruvate ameliorates intestinal epithelial barrier dysfunction in
endotoxemic mice and immunostimulated caco-2 enterocytic monolayers. J
Pharmacol Exp Ther. 2003 Jan;304(1):464-76). Upon dissolution in water
pyruvate undergoes condensation and cyclization type reactions resulting in a
variety of chemical species, some of which may be toxic (Montgomery CM, Webb
JL. Metabolic studies on heart mitochondria. II. The inhibitory action of
parapyruvate on the tricarboxylic acid cycle. J Biol Chem. 1956 Jul;221(1):359-
68). In order to overcome the shortcomings of pyruvate an ester derivative,
ethyl
pyruvate, was developed. Ethyl pyruvate will not undergo the condensation and
cyclization type reactions in water because of the ester protecting group.
Specific enzymes, such as esterases which are present in mammals are required
for the removal of the ethyl ester. Thus the use of ethyl pyruvate enhances
the
uptake of pyruvate by reducing the potential for condensation and cyclization.
Pyruvate is endogenously produced in cells as a result of the metabolism
of glucose by glycolysis. In situations where a cell has an adequate supply of
oxygen the pyruvate is converted into acetyl-coenzyme A, transported into the
mitochondria, and enters a series of reactions collectively known as the Krebs
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cycle. However, in situations of oxygen deficiency, often occurring in muscle
as
a result of extended periods of exercise, the pyruvate is converted into
lactate.
While pyruvate can be transported directly into the mitochondria, most of it
is
reduced to lactate in the cytosol, prior to transport. This reduction of
pyruvate
consumes a free proton from the cytoplasm and so acts as a buffer against
acidosis (Robergs R, Ghiasvand F, Parker D. Biochemistry of exercise-induced
metabolic acidosis. Am J Physiol Regul Integr Comp Physiol. 2004;287:R502-
16).
Lactate, resulting from the conversion of pyruvate, can be transported into
the mitochondria where it can be oxidized (Butz CE, McClelland GB, Brooks GA.
MCT1 confirmed in rat striated muscle mitochondria. J Appl Physiol. 2004
Sep;97(3):1059-66), or it can be exported out of the cell and taken up by
oxygen-
rich muscle cells. The transport of lactate into the mitochondria is
facilitated by
MCT proteins, which are proton-linked transporters, i.e. protons are co-
transported into the mitochondria with lactate (Roth DA, Brooks GA. Lactate
and
pyruvate transport is dominated by a pH gradient-sensitive carrier in rat
skeletal
muscle sarcolemmal vesicles. Arch Biochem Biophys. 1990 Jun:279(2):386-94).
Therefore, as cytostolic pH decreases as a result of ATP hydrolysis and
cytostolic concentrations of lactate increase as a result of ethyl pyruvate
administration, the co-transport of free protons and lactate out of the
cytosol is
increased.
It is herein understood by the inventors that inclusion of ethyl pyruvate in a
nutritional supplement will increase cellular levels of pyruvate. This
increased
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concentration of cellular pyruvate will facilitate greater conversion to
lactate, thus
greater consumption of free protons, and increased regeneration of NAD+. The
increased regeneration of NAD+ will facilitate greater levels of glycolytic
energy
production required by the working muscle and the increased consumption of
free cytosolic protons will buffer against acidosis resulting from the
increased
glycolytic energy production. Additionally, it is also understood by the
inventors
that increased levels of cytosolic lactate will increase the co-transport of
protons
and lactate into the mitochondria, thereby further buffering the cell against
acidosis as well as increasing the substrate inside the mitochondria available
to
fuel the citric acid cycle.
In an embodiment of the present invention, which is set forth in greater
detail in the examples below, the nutritional composition comprises ethyl
pyruvate. A serving of the nutritional composition comprises from about 0.0001
g
to about 0.75 g of ethyl pyruvate.
a-Hydroxyisocaproic acid (HICA)
a-Hydroxyisocaproic acid is an end product of the metabolism of the
branched chain amino acid, Leucine. Foods that are produced by fermentation,
such as some cheeses, may contain small amounts of HICA. HICA is a
reduction product of the a-keto acid analog of Leucine, a-ketoisocaproic acid
(KICA), and as such contributes to the free pools of branched chain amino
acids
(BCAA). HICA belongs to the group collectively known as branched chain amino
acid analogues.
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Branched chain amino acid analogues are essentially nitrogen-free amino
acids and may serve three roles in cases of nitrogen accumulation, 1)
providing the
dietary requirement of the corresponding branched-chain amino acid without
increasing nitrogen intake; 2) reducing the amount of nitrogen that must be
excreted
from the body; and 3) increasing levels of Leucine, which plays a key role in
protein
turnover and prevents wasting of lean body mass. It is important to note that
nitrogen accumulation can result from a number of situations including the
catabolism of proteins in muscle during exercise.
Since branched chain amino acid analogues may be reaminated back to
amino acid, e.g. HICA can be converted to KICA which can subsequently be
converted back to Leucine, they can act to provide the dietary requirements
for
BCAA without increasing level of ingested nitrogen (Boebek KP, Baker DH.
Comparative utilization of the a-keto and D- and L-a-hydroxy analogs of
Leucine,
Isoleucine and Valine by chicks and rats. J Nutr. 1982 Oct; 112(10):1929-39).
This
reamination will act to reduce accumulation of nitrogen in working cells,
which will
result in a reduction in the occurrence of delayed onset muscular soreness.
Administration of about 1.5 g of HICA daily after intense exercise for a
period
of 42 days (Karila T, Seppala T. a-Hydroxyisocaproic acid (HICA) - a Leucine
metabolite for muscle recovery following exercise. Oy Elmomed Ltd.) resulted
in a
statistically significant increase in total lean soft tissue mass.
Additionally it was
noted that subjects receiving HICA experienced little to no delayed onset
muscle
soreness. It is likely that this elimination of delayed onset
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muscle soreness is a result of inhibition of metalloproteinases, which are
responsible for degradation of the extracellular matrix during tissue
remodeling.
Additionally in high catabolic states, such as those induced by intensive
exercise, both a-keto acids and a-hydroxy acid metabolites of branched chain
amino acids may be oxidized for energy instead of the branched chain amino
acids themselves (Staten MA, Bier DM, Matthews DE. Regulation of valine
metabolism in man: a stable isotope study. Am J Clin Nutr. 1984 Dec;40(6):1224-
34). Using the deaminated metabolites, e.g. HICA, over the aminated forms,
e.g.
Leucine, will act to attenuate ammonia accumulation in working muscle. Also, a-
hydroxy acid analogues, like HICA, can be reaminated to yield the
corresponding
branched chain amino acids (Hoffer LJ, Taveroff A, Robitaille L, Marne OA,
Reimer ML. Alpha-keto and alpha-hydroxy branched-chain acid interrelationships
in normal humans. J Nutr. 1993 Sep;123(9):1513-21). Thus, oral administration
of at least one a-hydroxy branched-chain amino acid metabolite will act to
increase levels of the corresponding branched-chain amino acid present in
skeletal muscle.
Leucine is able to stimulate protein synthesis and inhibit protein
breakdown (Tischler ME, Desautels M, Goldberg AL. Does Leucine, leucyl-tRNA,
or some metabolite of Leucine regulate protein synthesis and degradation in
skeletal and cardiac muscle? J Biol Chem. 1982 Feb 25;257(4):1613-21), both of
which would be favorable in working muscle as they result in increased
skeletal
muscle growth and decreased recovery time.
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It is herein understood by the inventors that oral administration of HICA
will act to increase muscular concentrations of Leucine by acting as a
substitute
for Leucine in catabolism for energy as well as potentially being reaminated
to
form Leucine. Increased levels of Leucine will stimulate protein synthesis and
inhibit protein breakdown, thereby inducing an anabolically-favorable state
for the
cell. Additionally, it is herein understood by the inventors that oral
administration
of HIMVA and HIVA will act to increase muscular concentrations of Isoleucine
and Valine, respectively, by at least the mechanisms discussed above.
In an embodiment of the present invention, which is set forth in greater
detail in the examples below, the nutritional composition comprises a-
hydroxyisocaproic acid. A serving of the nutritional composition comprises
from
about 0.00001 g to about 0.75 g of a-hydroxyisocaproic acid.
In a further embodiment of the present invention, which is set forth in
greater detail in the examples below, the nutritional composition comprises a-
hydroxy f-methylvaleric acid. A serving of the nutritional composition
comprises
from about 0.00001 g to about 0.75 g of a-hydroxy-,B-methylvaleric acid.
In an embodiment of the present invention, which is set forth in greater
detail in the examples below, the nutritional composition comprises a-hydroxy-
isovaleric acid. A serving of the nutritional composition comprises from about
0.00001 g to about 0.75 g of a-hydroxy-isovaleric acid.
In various embodiments of the present invention, which are set forth in
greater detail in the examples below, the nutritional supplement comprises
ethyl
pyruvate and at least one a-hydroxy branched-chain amino acid metabolite. The
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nutritional supplement is provided in any acceptable and suitable oral dosage
form as known in the art. A synergistic anabolically-favorable state for the
cell,
via substantially simultaneously maintaining physiological pH levels and
increasing cellular concentrations of branched-chain amino acids, is induced
and
carried out in an individual by administration of the composition of the
present
invention.
The nutritional supplement of the present invention may be administered
in a dosage form having controlled release characteristics, e.g. time-release.
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 constituent release strategies
extend
the period of bioavailability or target a specific time window for optimal
bioavailability. Advantageously the nutritional supplement may be administered
in the form of a multi-compartment capsule which combines both immediate
release and time-release characteristics. Individual components of the
nutritional
supplement may be contained in differential compartments of such a capsule
such that specific components are released rapidly while others are time-
dependently released. Alternatively, a uniform mix of the various components
of
the present invention may be divided into both immediate release and time-
release compartments to provide a multi-phasic release profile.
While not wishing to be bound by theory, the present invention is
comprised of components which act to attenuate acidosis in working muscle by
increasing the conversion of pyruvate to lactate, which consumes free
cytostolic
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protons (H+). Additionally, increased concentrations of lactate, which result,
will
increase the co-transport of lactate and cytostolic H+ into the mitochondria,
thereby decreasing the concentration of cytostolic protons and increasing
substrates in the mitochondria which are available to fuel the citric acid
cycle.
Both of the aforementioned mechanisms will enhance the buffering ability of
the
cell. Since decreased cellular pH has been linked to cell damage (Bevilacqua
L,
Ramsey JJ, Hagopian K, Weindruch R, Harper ME. Effects of short- and
medium-term calorie restriction on muscle mitochondrial proton leak and
reactive
oxygen species production. Am J Physiol Endocrinol Metab. 2004
May;286(5):E852-61 (Abstract)) leading to degradation, attenuation of pH
decreases would be anti-catabolic and as such would act to induce an
anabolically-favorable state for the cell.
Additionally, the present invention comprises components which have
been shown to increase levels of branched-chain amino acids. It is herein
understood by the inventors that inclusion of HICA in the nutritional
supplement
will act to increase muscular concentrations of Leucine by acting as a
substitute
for Leucine in catabolism for energy as well as potentially being reaminated
to
form Leucine. Increased levels of Leucine will stimulate protein synthesis and
inhibit protein breakdown, thereby inducing an anabolically-favorable state
for the
cell.
Further to the aforementioned functions, it is herein understood that
administration of ethyl pyruvate and HICA concomitantly in a single serving of
the
nutritional supplement act substantially simultaneously to induce an
anabolically-
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favorable state in cells. Administration of ethyl pyruvate increases cellular
concentrations of lactate which are preferentially utilized for energy,
thereby sparing
the concentrations of BCAA, such as Leucine. In a manner similar to ethyl
pyruvate,
HICA will spare the concentrations of BCAA, however, HICA has the additional
capability of conversion to Leucine, thereby not only conserving Leucine, but
also
acting to increase the concentrations of Leucine. Therefore, administration of
ethyl
pyruvate and HICA together will conserve and increase Leucine concentrations,
wherein an anabolically-favorable state for the cell is induced.
Additional embodiments of the present invention may also include portions of
the composition as fine-milled ingredients. U.S. Non-Provisional Patent
Publication
20070202166 entitled "Method for Increasing the Rate and Consistency of
Bioavailability of Supplemental Dietary Ingredients" filed Feb 21, 2007
discloses a
method of increasing the rate of bioavailability following oral administration
of
components comprising supplemental dietary compositions by the process of
particle-milling.
Furthermore, additional embodiments of the present invention may be
incorporated into specific controlled-release solid dosage forms. U.S. Non-
Provisional Patent Publication 20070202165 entitled "Method for a Supplemental
Dietary Composition Having a Multi-Phase Dissolution Profile" filed Feb 21,
2007
discloses a method of achieving a solid oral dosage form with multiple
dissolution
characteristics for the release of active ingredients.
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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
beverage, a ready-to-eat bar or drink product, a capsule, a liquid capsule, a
tablet, a caplet, or as a dietary gel. The preferred dosage forms of the
present
invention are as a caplet or as a liquid capsule.
Furthermore, the dosage form of the nutritional supplement may be
provided in accordance with customary processing techniques for herbal and
nutritional supplements in any of the forms mentioned above. Additionally, the
nutritional supplement set forth in the example embodiment herein disclosed
may
contain any appropriate number and type of excipients, as is well known in the
art. By way of ingestion of the composition of the present invention, a method
for
inducing an anabolically-favorable state for the cell by substantially
simultaneously maintaining blood and muscle physiological pH levels and
increasing cellular concentrations of Leucine, is provided. The method of the
present invention comprises at least the step of administering to an
individual a
therapeutically acceptable amount of the composition of the present invention.
Although the following example illustrates the practice of the present
invention in one of its embodiments, the example should not be construed as
limiting the scope of the invention. Other embodiments will be apparent to one
of
skill in the art from consideration of the specifications and example.
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Examples
Example 1
A nutritional supplement comprising the following ingredients per serving is
prepared for consumption as a caplet three times daily prior to meals:
from about 0.0001 g to about 0.75 g of ethyl pyruvate and from about 0.00001 g
to about 0.75 g of a-hydroxyisocaproic acid (HICA).
Example 2
A nutritional supplement comprising the following ingredients per serving is
prepared for consumption as a caplet three times daily prior to meals:
about 0.001 g of ethyl pyruvate and about 0.0001 g of a-hydroxyisocaproic acid
(HICA).
Example 3
A nutritional supplement comprising the following ingredients per serving is
prepared for consumption as a time-release multi-compartmented capsule twice
daily prior to meals, preferably once before the first meal and once before
the last
meal of the day:
about 0.005 g of ethyl pyruvate and about 0.0005 g of a-hydroxyisocaproic acid
(HICA).
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Example 4
A nutritional supplement comprising the following ingredients per serving is
prepared for consumption as a capsule to be taken once daily prior to
exercise:
about 0.01 g of ethyl pyruvate, about 0.001 g of a-hydroxyisocaproic acid
(HICA),
about 0.001 g of a-hydroxyfl-methylvaleric acid (HIMVA), and about 0.001 g of
a-hydroxy-isovaleric acid (HIVA).
Example 5
A nutritional supplement comprising the following ingredients per serving is
prepared for consumption as a caplet to be taken once daily following
exercise:
about 0.01 g of ethyl pyruvate, about 0.001 g of a-hydroxyisocaproic acid
(HICA),
about 0.001 g of a-hydroxy-,B-methylvaleric acid (HIMVA), and about 0.001 g of
a-hydroxy-isovaleric acid (HIVA).
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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.
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