Title: Composition and method
for treating age-related disorders
United States Patent: 7,033,612
Issued: April 25, 2006
Inventors: Kang; David S.
(16 Forest Gate Cir., Oakbrook, IL 60523); Kang; Chunghee Kimberly (16
Forest Gate Cir., Oakbrook, IL 60523)
Appl. No.: 336150
Filed: January 3, 2003
There is provided a composition for the
prevention and treatment of age-related physical and mental disorders that
includes phospholipids that have been extracted from chick embryos aged
between about 6 days old and about 14 days old. Also provided is a method
for extracting these phospholipids by incubating them for between about 6
days to about 14 days. The chick embryos are then prepared for chemical
extraction, and the lipids are extracted from the chick embryos. Further
provided is a method for preventing and treating age-related physical and
mental disorders, in human subjects in need thereof, by administering a
sufficient dosage of a composition including phospholipids extracted from
chick embryos incubated for between about 6 days and about 14 days to a
BRIEF SUMMARY OF THE
According to one aspect of the present
invention, there is provided a composition for the prevention and
treatment of age-related physical and mental disorders. This composition
includes phospholipids that have been extracted from chick embryos aged
between about 6 days old and about 14 days old.
According to another aspect of the present invention, there is provided a
method for extracting phospholipids from chick embryos for use in a
composition for alleviating age-related physical and mental disorders. The
fertilized eggs are incubated for between about 6 days to about 14 days.
The chick embryos are separated and the lipids of chick embryos are
extracted by organic solvents. Phospholipids are separated from solvents
and residual water by evaporation and freeze-drying.
According to yet another aspect of the present invention, there is
provided a method for using a composition for preventing and treating
age-related physical and mental disorders, in a human subject in need
thereof. A dosage of a composition including phospholipids extracted from
chick embryos incubated for between about 6 days and about 14 days is
administered to the subject. The dosage is of a sufficient amount to
increase physical endurance.
According to still another aspect of the present invention there is
provided a method for treating age-related physical and mental disorders,
in a human subject in need thereof. A dosage of the composition including
phospholipids extracted from chick embryos incubated for between about 6
days and about 14 days is administered to the patient. The dosage is of a
sufficient amount to increase physical endurance.
OF THE INVENTION
The present invention relates generally
to the field of therapeutic uses of embryonic phospholipids, particularly
plasmalogens of phosphoglycerides prepared from chicken embryo tissues,
for prevention and treatment of age-related disorders. The present
invention provides a composition for preventing and treating these
age-related physical and cognitive disorders that comprises a phospholipid
extract collected from 6 to 14 day-old chick embryos. The present
invention comprises pharmaceutical compositions containing early chick
embryo phospholipids, particularly those including plasmalogens, mixed
into an inert, nontoxic carrier for oral or sublingual administration, and
methods of preparing and administering these compositions.
There are generally two groups of phospholipids present in embryonic and
post-natal chicken brains. The first group comprises phosphatidylcholine,
phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol. The
second group comprises plasmalogens of choline, ethanolamine and serine.
This second group also includes sphingomyelin, and ethanolamine ether
phosphatides. In young chicken embryos, the phospholipids of the first
group dominate the chemical makeup of the embryos. Up until chick embryos
are about 16 days old, the phospholipids of the first group are
synthesized very rapidly, while there is only a slight increase in the
amount of phospholipids of the second group. When chick embryos are aged
between about 16 and about 20 days old, the phospholipids of the second
group begin to rapidly synthesize as well, bringing the level of
phospholipids of the second group close to that of the first group. For
the purposes of preventing and treating age-related illnesses, the
preferred embryo age is between about 6 and about 14 days old; more
preferably, the embryo age is between about 9 and about 10 days old.
Phospholipids in early to mid-stage embryos—those aged from about 6 to
about 14 days—contain strikingly different phosphoglycerides than
late-stage embryos. These mid-stage phosphogeycerides contain alkenyl and
acyl groups not typically present in late stage phosphoglycerides. For
example, mid-stage phosphoglycerides comprise ethanolamine plasmalogens
with short chain lauric or myristic ester linkages and palmityl ether
Plasmalogens and phospholipids extracted from mid-stage embryos play an
important role in the prevention and treatment of age-related disorders.
For example, the presence of plasmalogens in substantial amounts in the
inner layer of biological membranes is well-documented, indicating that
these lipids are involved in signal transduction following binding
regulatory mechanisms within these membranes. It is believed that there is
specific participation by ether lipids in the chemiosmotic process,
similar to the participation of such inner mitochondrial membrane
components as coenzyme Q, NADH, FMN and cytochrome a 3. It is
also believed that membrane lipid composition is strongly influenced by
lipid shape, and that the observed changes in lipid composition serve to
stabilize the bilayer arrangement of cellular and subcellular membranes.
All these factors can strengthen the membranes of the cell's surface, as
well as those of subcellular organelles, reducing the effects of
Plasmalogens also are potential protectors against the oxidation of LDL
proteins. About 3% to about 5% of total LDL phospholipids are plasmalogens.
In vitro oxidation of LDL using an oxidizing agent induces a selective
reduction of plasmalogen subgroups in phosphatidylcholine and
phosphatidylethanolamine. VLDL and LDL plasmalogens were oxidized to a
greater extent (99% decrease) compared to HDL plasmalogens (35%),
suggesting that plasmalogens play a protecting role with respect to
atherogenesis, the build-up of fats and fatty acids in the blood stream.
This also contributes to the regulation the blood pressure, which is also
affected by age.
The content of cholesterol and phospholipids in 8 and 16 day old chick
embryo fibroblasts has also been compared. Cholesterol content did not
change, whereas total phospholipids decreased with time change, resulting
in an increase of the cholesterol to phospholipid ratio. Further, the
fatty acid composition of phosphatidylcholine and phosphatidylethanolamine
in these fibroblasts showed unique features. For example, the content of
myristic acid was depleted by more than two-thirds from the 8 to 16 day
old fibroblast. This indicates a change in the fluidity of membranes
during development, a change that aids in keeping cells relatively strong
and helps to assuage age-related conditions.
To prepare a composition for preventing and treating age-related
conditions, chick embryos are selected that are aged between about 6 days
and about 14 days; preferably, between about 9 days and about 10 days.
These embryos have been preferably incubated for the duration of aging at
a temperature between about 90° F. to about 100° F., preferably between
about 95° F. to about 100° F., more preferably between about 99° F. to
about 100° F. The embryos are then stored until use by immediately
freezing the embryos and storing them in a freezer at a temperature
between about -70° C. to about -90° C., preferably between about -75° C.
to about -85° C., more preferably between about -80° C. to about -85° C.
To prepare the frozen embryos for use, they are first thawed, and then
briefly minced with a blender. The minced embryo pieces are then
centrifuged by any method known to one of skill in the art. The
centrifugation is performed in order to separate out the liquid portions
of the embryos. Any residual water in the solid phase is then removed by
further centrifugation methods after mixing with ethanol. The residual
water may also be removed by freeze-drying methods, warm air currents, or
any other dessication method known to one of skill in the art. If
centrifugation methods are used with ethanol, acetone may be added to the
ethanol to improve drying properties. In a large scale preparation, minced
embryo tissues may be dried in an evaporator having a partial vacuum. In
such an evaporator, the vacuum pressure should be maintained such that the
boiling point of the minced tissues is between about 50° C. and about 60°
C. Preferably, the vacuum pressure is maintained such that the boiling
point is between about 54° C. and about 57° C. If necessary, a warm air
current is used to further dry the condensed minced embryo mixtures. The
air current should be maintained at about 180° C. to about 200° C.,
preferably at about 188° C. to about 192° C., for best results in drying
the embryo pieces without chemically destroying the biological compounds.
The minced, dried embryo tissues are then mixed with suitable organic
solvents, such as a hexane-ethanol solution, wherein the lipids in the
minced embryo tissues are extracted into the organic solvent. Preferably,
the lipids are extracted under stirring. As a result, a lipid extract is
removed from the liquid phase, and the lipid extract is temporarily set
aside. The liquid phase left after lipid extraction is then concentrated
to about 5% to about 15% of the original volume, preferably concentrated
to about 9% to about 11% of the original volume. This may be done by any
method known in the art, but is preferably performed by using a flash
evaporator at less than about 40° C. to about 50° C., more preferably at
about 43° C. to about 46° C.
The liquid phase is then separated into an aqueous and organic layer. The
organic layer is then further concentrated to create a phospholipid
extract. In order to eliminate any residual water and organic solvents
from the phospholipid extract, the extracted phospholipids can be
lyophilized after mixing them with a portion of inactive ingredients. For
example, an alimentary antioxidizing agent and isoleucine can be added to
the lyophilized mixture. The phospholipid extract is preferably
lyophilized by a standard freeze-drying method, but may be purified by any
method known to those of skill in the art.
A small portion of the phospholipid extract is set aside and saved for
quality analysis of the resulting lipids in the composition. Analysis can
be performed by thin layer chromatography, high performance liquid
chromatography (HPLC), gas chromatography, NMR spectrometry, or any
combination of these techniques. Typically, NMR spectroscopy shows that as
a result of this method, the phospholipid extract contains
phosphatidylcholine comprising about 50% to about 70% of the total
phospholipids, more preferably comprising about 55% to about 66% of the
total phospholipids. Plasmalogens of phosphatidylcholine are generally not
detected in any appreciable amount in the phospholipid extract resulting
from this method. Phosphatidylethanolamine and its plasmalogen comprises
about 20% to about 30% of the total phospholipids in the phospholipid
extract, more preferably 23% to about 26% of the total phospholipids.
Further analysis also shows that the major fatty acids found in the
phospholipid extract include C18:0 and C16:0 (from
phosphatidylethanolamine) and C16:0, C20:4n6, C16:1, C22:6n3, and C18:1
(from phosphatidylcholine). The chemical nomenclature used above indicates
the length of the fatty acid (indicated by the number directly after the
"C"), as well as the number of double bonds along its backbone (indicated
by the number directly after the colon) and the position of these double
bonds (indicated by the number after an "n").
To eliminate any residual water and organic solvent, the phospholipid
extracts are lyophilized after mixing them with an alimentary
antioxidizing agent and amino acids. Amino acids are a basic building
block of the human body, and supplements including these amino acids serve
to help replenish those levels. Any amino acid that is hydrophobic may be
used in this compound. Common amino acids, such as lysine, leucine,
isoleucine and cyteine may be used. More preferably, isoleucine is used. A
particularly hydrophobic amino acid, isoleucine is often found
concentrated in muscle tissues. In particular, isoleucine aids the
formation of hemoglobin in the human body. Isoleucine also serves to
stabilize and regulate blood sugar and energy levels throughout the body.
Further, the phospholipid extract can be lyophilized or otherwise admixed
with an antioxidant. Antioxidant compounds are those that reduce the
oxidization of the cellular structure. This slows the cellular aging
process, and keeps cells stronger. In the present mixture, the addition of
antioxidants serves to help prevent the oxidation of phospholipids.
Preferred antioxidants for use in the composition comprise vitamin E, as
well as its derivatives, and vitamin C, as well as its derivatives. More
preferably, vitamin E and ascorbyl palmitate, a vitamin C derivative, are
used. Vitamin E, a fat-soluble, naturally occurring vitamin, has
particularly good anti-oxidant properties. Vitamin E has been linked to a
reduction in the rate of coronary diseases. Vitamin C serves not only to
prevent cellular oxidation, but also to aid the synthesis of collagen in
the body, which itself helps to repair aged skin and muscle tissues.
Ascorbyl palmitate has been shown to be a particularly effective
derivative of vitamin C in supplements. These antioxidants may also
replace the amino acids in an embodiment of the present invention.
Additionally, a binding element is preferably added to the phospholipid
extract. The binding element serves to hold the ingredients of the
composition together. Any stearate-based compound safe for human
consumption may be used, such as calcium stearate and zinc stearate.
However, a preferred binding element is magnesium stearate. A derivative
of magnesium, this chemical compound is often used as a food additive to
promote binding in food products.
Polysaccharides, which are carbohydrate-based starches comprising a
plurality of sugar molecules, also help to hold the ingredients of the
composition together. Any polysaccharide may be used; however, preferred
polysaccharides in the composition include lactose, cellulose, and
fructose. More preferably, lactose and cellulose are used.
Other ingredients may also be added to improve the taste or consistency of
the final formulation, as is known in the art. For example, a flavoring
agent may be added to the composition. Such a flavoring agent would
improve the taste of the composition upon administration, letting it taste
more like mint, fruit, or any taste preferred by the consumer. Use of a
flavoring agent is preferable in a composition to be administered in a
liquid form, but may also be used in a solid form composition.
After preparing the phospholipid extract, the chosen ingredients are then
mixed together to create doses of the final composition. This composition
may be in a liquid form or a solid capsule form. The composition may be
administered in any form known to those in the art; for example, a solid
capsule may be a film-coated tablet or a soft gelatin tablet. As a further
example, a liquid composition may be administered as a liquid elixir or as
a sublingual or oral spray. While the form of administration may change,
the dosage should be calculated such that a single dosage comprises about
40 mg to about 100 mg of phospholipid extract. More preferably, a single
dosage comprises about 50 mg to about 80 mg of phospholipid extract.
Claim 1 of 31 Claims
1. A composition for
alleviating age-related physical and mental disorders, comprising
phospholipids extracted from chick embryos aged between about 6 days old and
about 14 days old, wherein said phospholipids comprise about 50% to about
70% by weight of phosphatidylcholine and about 20% to about 30% by weight of
phosphatidylethanolamine and its plasmalogens.
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