U.S. patent application number 15/455006 was filed with the patent office on 2017-08-24 for glycogen or polysaccharide storage disease treatment method.
The applicant listed for this patent is Baylor Research Institute. Invention is credited to Charles R. ROE.
Application Number | 20170239205 15/455006 |
Document ID | / |
Family ID | 35787579 |
Filed Date | 2017-08-24 |
United States Patent
Application |
20170239205 |
Kind Code |
A1 |
ROE; Charles R. |
August 24, 2017 |
GLYCOGEN OR POLYSACCHARIDE STORAGE DISEASE TREATMENT METHOD
Abstract
A method for treating glycogen storage disease by administering
an effective amount of a composition that includes ketogenic odd
carbon fatty acids that ameliorate the symptoms of these
diseases.
Inventors: |
ROE; Charles R.; (Rockwell,
TX) |
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Applicant: |
Name |
City |
State |
Country |
Type |
Baylor Research Institute |
Dallas |
TX |
US |
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|
Family ID: |
35787579 |
Appl. No.: |
15/455006 |
Filed: |
March 9, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14883981 |
Oct 15, 2015 |
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15455006 |
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14214317 |
Mar 14, 2014 |
9186344 |
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14883981 |
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13343578 |
Jan 4, 2012 |
8697748 |
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14214317 |
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11172693 |
Jul 1, 2005 |
8106093 |
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13343578 |
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60585502 |
Jul 2, 2004 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 9/0053 20130101;
A61K 38/47 20130101; A23L 33/115 20160801; A23K 50/80 20160501;
A61K 31/19 20130101; A23K 20/174 20160501; A61K 31/202 20130101;
C12Y 302/0102 20130101; A23L 33/10 20160801; A23L 33/12 20160801;
A61K 31/225 20130101; A23K 50/75 20160501; A23L 33/16 20160801;
A61K 31/20 20130101; A23K 50/10 20160501; A61P 3/00 20180101; A23L
33/175 20160801; A61K 9/0095 20130101; A23K 20/158 20160501; A61K
31/23 20130101; A23L 2/52 20130101; A61K 31/22 20130101; A23V
2002/00 20130101 |
International
Class: |
A61K 31/225 20060101
A61K031/225; A23L 33/115 20060101 A23L033/115; A23L 33/16 20060101
A23L033/16; A61K 9/00 20060101 A61K009/00 |
Claims
1-100. (canceled)
101. A method of treating a fatty acid oxidation disorder (FOD) in
a subject in need thereof, wherein the method comprises
administering to the subject a composition comprising an active
agent, wherein the active agent consists essentially of a
triheptanoin, and wherein the composition has an acid value of 0.1
or less mg KOH per gram and a hydroxyl value of 2.8 or less mg KOH
per gram.
102. The method of claim 101, wherein the composition is
administered in a range of 1 to 4 grams of triheptanoin per
kilogram body weight per day.
103. The method of claim 101, wherein the composition is
administered in a range of 0.1 to 2 grams of triheptanoin per
kilogram body weight per day.
104. The method of claim 101, wherein the composition is in a
dosage unit, wherein the dosage unit comprises at least 15 grams of
the triheptanoin.
105. The method of claim 104, wherein the dosage unit comprises 15
to 75 grams of the triheptanoin.
106. The method of claim 101, wherein the composition is provided
in an amount that provides 15% to 40% of the daily dietary calories
of the subject.
107. The method of claim 101, wherein the composition is provided
in an amount that provides 20% to 35% of the daily dietary calories
of the subject.
108. The method of claim 101, wherein the composition is provided
in an amount that provides 25% to 35% of the daily dietary calories
of the subject.
109. The method of claim 101, wherein the composition is provided
as a nutritional supplement.
110. The method of claim 101, wherein the composition is provided
as a dietary formulation.
111. The method of claim 101, wherein the composition is a
pharmaceutical composition.
112. The method of claim 101, wherein the composition is
administered parenterally.
113. The method of claim 101, wherein the composition is
administered enterally.
114. The method of claim 101, wherein the composition is
administered orally.
115. The method of claim 101, wherein the FOD is selected from
carnitine palmitoyl transferase I (CPT I) deficiency,
carnitine-acylcarnitine translocase (CACT) deficiency, carnitine
palmitoyl transferase I (CPT II) deficiency, very long chain
acyl-CoA dehydrogenase (VLCAD) deficiency, trifunctional protein
(TFP) deficiency, long chain acyl 3-hydroxy CoA dehydrogenase
(LCHAD) deficiency and short chain acyl CoA dehydrogenase (SCAD)
deficiency.
116. The method of claim 114, wherein the FOD is CPT II
deficiency.
117. The method of claim 114, wherein the FOD is VLCAD
deficiency.
118. The method of claim 114, wherein the FOD is LCHAD
deficiency.
119. The method of claim 114, wherein the FOD is TFP
deficiency.
120. The method of claim 114, wherein the FOD is CACT deficiency.
Description
[0001] This application claims priority to U.S. Provisional Patent
Application Ser. No. 60/585,502 filed Jun. 2, 2004, the entire
contents of which are incorporated herein by reference. Without
limiting the scope of the invention, its background is described in
connection with polysaccharide storage diseases.
TECHNICAL FIELD OF INVENTION
[0002] This invention relates to the treatment of glycogen or
polysaccharide storage diseases affecting humans and other animals,
and more particularly, to formulations that ameliorate the symptoms
of these diseases.
BACKGROUND OF THE INVENTION
[0003] Glycogen-storage disease type II ("GSD II" or "Pompe's
disease"), is a genetic disorder in humans resulting from the
deficiency of acid alpha-glucosidase ("acid maltase"), a lysosomal
hydrolase enzyme. The disease is characterized by the abnormal
accumulation of glycogen in the lysosomes. (Hirschhorn, R and
Reuser, A J J: Glycogen Storage Disease Type II: Acid
.alpha.-Glucosidase Deficiency. In: The Metabolic and Molecular
Bases of Inherited Diseases, 8th edition, Chapter 135, pp
3389-3420, McGraw-Hill, 2001.). Polysaccharide storage diseases
have also been reported in other animals, particularly in horses,
cattle, and sheep.
[0004] With respect to humans, three major forms of GSD II have
been described: infantile, juvenile, and adult-onset. See J.
Ibrahim, et al., Glycogen Storage Disease Type II,
http://www.emedicine.com/PED/topic1866.htm (2003). The infantile
form of II is described as presenting by 6 months of age and
characterized by involvement of cardiac, skeletal and respiratory
muscles with rapid progression to death by respiratory and cardiac
failure. These inherited conditions are due to an enzyme-deficiency
that occurs in the human population at about 1 in 14,000 to 1 in
60,000 live births. The infantile or neonatal form of the disease
usually results in death by about twelve to eighteen months of
age.
[0005] The juvenile (intermediate) form includes infants and
children older than 6 months who present with weakness but
generally have no cardiac disease. Adult-onset GSD II is slowly
progressive and involves progressive muscle weakness that affects
the accessory muscles of respiration and finally leads to
respiratory failure and death. This form of the disease usually
does not involve the heart. The adult-onset form of the disease may
appear in the second or third decade of life, and as late as the
sixth decade. The disease results in loss of weight and muscle
mass. With the loss of muscle mass comes difficulty in breathing
because the muscles have difficulty powering the breathing
mechanism. The deficient enzyme, acid .alpha.-glucosidase, is not
required for the vast majority of cellular glycogen because the
main pathway for glycogen degradation is not deficient in GSD II
disease, energy production is not impaired, and hypoglycemia does
not occur. The main issue of deficiency of acid alpha-glucosidase
enzymatic activity is the accumulation of structurally normal
glycogen in lysosomes and cytoplasm in affected individuals.
Excessive glycogen storage within lysosomes interrupts the normal
functioning of other organelles and leads to cellular injury and
dysfunction of the entire organ involved.
[0006] Presently, there is neither a cure for the disease, nor
alteration of the clinical course of expected fatality, although
some relief has been realized temporarily with high protein diets
and with treatment for cardiac and respiratory failure.
Alternatively, enzyme replacement therapy with recombinant human
acid alpha-glucosidase, (rhGAA), an investigational enzyme
replacement therapy for Pompe disease is now in clinical trials
(www.genzyme.com and www.pompe.com).
[0007] Glycogen and other polysaccharide storage diseases also
affect animals other than humans. For example, muscle disease in
draft horses has been known for over 100 years, and in 1992 was
correlated with polysaccharide storage diseases. Such disorders are
commonly referred to as
[0008] "Monday Morning Disease", "tying" or "locking up" or simply
severe muscle disease have been reported. Draft horses which worked
hard six days a week but were given a high grain diet and a day of
rest on Sunday were found to be prone to massive muscle injury on
Monday. Recently, the polysaccharide storage diseases have been
found to be related to problems with giving a draft horse general
anesthesia. In addition, glycogen branching enzyme deficiency has
been reported as causing muscle weakness in Quarter Horses and
related breeds. It is reportedly a separate disorder from
polysaccharide storage myopathy and is a deficiency in the enzyme
necessary for the formation of normal glycogen. S. Valberg,
"Glycogen Storage Disorders of Quarter Horse Foals," J. Vet.
Intern. Med. 15(6): 572-80 (2001).
[0009] A dietary solution has been suggested for the polysaccharide
storage diseases using a diet with reduced starches and sugars and
added fat as an alternative energy source. Providing fats as
opposed to carbohydrates as an alternative source of energy to
relieve tying up in horses has also been proposed. A subset of
horses with chronic exertional rhabdomyolyis have an abnormal
accumulation of glycogen (polysaccharide) stores in their muscles
and one of the preventative measures suggested is feeding diets
without grains and adding a fat supplement to maintain low blood
glucose and insulin concentrations. L. Gray, "Polysaccharide
Storage Myopathy (Glycogen Storage Disease)," Horse Previews, July
2002. These diets provide only partial relief, if at all, in these
affected animals.
SUMMARY OF THE INVENTION
[0010] It has now been found that administering odd carbon fatty
acids and/or ketones to humans and other animals can treat and
ameliorate the effects of glycogen or polysaccharide storage
diseases. More particularly, the present invention includes
compositions, methods, feeds, diets, additives and the like for the
treatment of polysaccharide storage diseases.
[0011] For example, the present invention includes a pharmaceutical
composition for treating a glycogen storage disease that includes a
pharmaceutically effective amount of an odd carbon fatty acid that
is at least partially water-soluble for the treatment of the
glycogen storage disease. The composition may also include a
carrier, a diluent, e.g., a lipophilic diluent and/or an
emulsifier. Non-limiting examples of useful emulsifiers include
Imwitor 370, Imwitor 375, Imwitor 377, Imwitor 380 and Imwitor 829.
In one embodiment, the odd carbon fatty acid is unneutralized and
may be a C15, C7, C5 and mixtures or combinations thereof. For
example, the odd carbon fatty acid may be 3-hydroxypentanoate,
3-ketopentanoate, tri-heptanoid, n-heptanoic acid and mixtures and
combinations thereof. The compositions of the present invention may
be provided to a patient suspected of having a Glycogen Storage
Disease, such as Pompe's disease. The patient may be suspected of
having an anapleurotic disease, that is, a disease with symptoms
that may be ameliorated with anapleurotic precursors that serve
have therapeutic effects.
[0012] For example, the composition of the present invention may be
an odd carbon fatty acid that is adapted for a dosage of between
0.1, 1, 1.5, 2, 3 or even 4 gr/kg/day. One example of an odd carbon
fatty acid for use with patients may have an acid value of 0.1 or
less mg KOH/gr, a hydroxyl value of 2.8 or less mg KOH/gr. The odd
carbon fatty acid may have a purity of at least about 98 percent,
e.g., triheptanoate that is at least about 97% pure.
[0013] The present invention also includes a method of treating a
patient suffering from Pompe's disease by administering to a
patient in need thereof a composition that with a human recombinant
.alpha.-glucosidase in a pharmaceutically acceptable carrier, in an
amount insufficient to treat the disease and concurrently providing
the patient with a pharmaceutically effective amount of an odd
carbon fatty acid to reduce the amount of the recombinant
.alpha.-glucosidase necessary to treat the patient. The amount of
human recombinant .alpha.-glucosidase may be from about 0.01 to
about 100 milligrams per 100 kilograms of patient per month. The
.alpha.-glucosidase and pharmaceutically effective amount of an odd
carbon fatty acid may be provided in a single-dose, e.g.,
intravenously. The odd carbon fatty acid may dosed at between 1 to
2 gr/kg/day and/or be 30 to 40 percent of total daily Kcalories.
Generally, a patient's total caloric intake is between about 1,200
to 3,000 calories per day.
[0014] Another embodiment of the present invention is a method of
treating a disease in a mammal resulting from deficiencies of fatty
acid oxidaase disease by administering to the mammal a
therapeutically effective amount of a pharmaceutical composition
that includes a recombinant glycosylated enzymatically active
.alpha.-glucosidase A or an enzymatically active fragment thereof
and a pharmaceutically effective amount of an odd carbon fatty
acid. By providing the patient with a supplemental source of
anapleurotic carbon sources, the effective amount recombinant
glycosylated enzymatically active .alpha.-glucosidase A or an
enzymatically active fragment needed to treat a mammal is reduced
by between about 20 and 80 percent. The mammal may be selected from
humans, dogs, cats, horses, hamsters, rats, mice, sheep, goats and
pigs. Depending on the course of treatment, specific disease,
extent of the disease and other biochemical, metabolic and
physiologic factors, the skilled artisan may be able to reduce the
amount and total number of daily doses of pharmaceutical
composition by half or less. The pharmaceutical composition may be
administered orally, intravenously, intramuscularly, intranasally,
intradermally, intraperitoneally, subcutaneously and/or as a
suppository or by any other method known to those of skill in the
pharmaceutical arts. The .alpha.-glucosidase A may further include
a pharmacologically acceptable carrier, e.g., polyethylene glycol
(PEG) and the amount of odd carbon fatty acid is sufficient to be
detectable in serum as free heptanoate and/or metabolites thereof.
By providing additional energetic support, the recombinant
glycosylated enzymatically active .alpha.-glucosidase A or an
enzymatically active fragment thereof is provided in a sub-optimal
amount. The effective amount of the odd carbon fatty acid includes
a nutritionally effective amount, that is, one that provides a
measurable amount of change in a patient, e.g., decreased fatigue,
increased energy, increased protein sparing and the like.
[0015] The present invention also includes a method of treating a
glycogen storage disease by administering to a patient in need
thereof a therapeutically effective amount of a pharmaceutical
composition that includes an odd-chain fatty acid or ketone. The
pharmaceutical composition is administered, e.g., at a daily oral
administration dosage of between about 0.5 to about 2.0 grams, per
day, per kilogram of patient weight. Useful dosage forms include
those adapted for oral administration, e.g., powder, tablet,
gelatin, gelcap, capsule, soft-gel, chewable or liquid form. For
human patient's, therapeutically effective amount are between about
20 and 40% of the patient's daily caloric intake. The compositions
may also be used for the treatment of muscle weakness by
administering to a patient a therapeutically effective amount of a
pharmaceutical composition with an odd-chain fatty acid or ketone,
wherein the patient's muscle weakness is reduced and pulmonary
function is increased. Muscle weakness may be the result of an
underlying disease or may be caused by acute muscle usage, e.g.,
athletes, mountain climbers, spelunking, military operations,
maneuvers, exercises and the like. The present invention will be
particularly useful for chronic muscle weakness. The present
invention may also be used in a treatment for urgency in urination
and defecation by administering to a patient a therapeutically
effective amount of a pharmaceutical composition with an effective
amount of an odd-chain fatty acid or ketone that reduces the
urgency to urinate and/or defecate.
[0016] Yet another method of the present invention includes a
method for protein sparing in which a therapeutically effective
amount of a pharmaceutical composition that includes an odd-chain
fatty acid or ketone is administered to a patient in need of
protein sparing. The composition may further includes one or more
vitamins, minerals, amino acids, lipids, nucleic acids, co-factors,
pro-vitamins, and combinations of mixtures thereof, e.g., a
nutritionally effective amount of branched amino acids.
[0017] The present invention may be formulated into immediate
release product with an at least partially water-soluble ketogenic
odd-chain fatty acid in an immediate release form that becomes
immediately bioavailable in a subject. Bioavailability in a subject
may be determined by serum levels of odd-chain fatty acids, organic
salts, acylcarnitines and carnitine levels from, e.g., dried blood
spots (dbs), saliva, urine, tears, sweat, plasma, amniotic or
cerebro-spinal fluid and the like. The product may be adapted for
parenteral, intravenous, oral, intramuscular, intraaortal,
intrahepatic, intragastric, intranasal, intrapulmonary,
intraperitoneal, subcutaneous, rectal, vaginal, intraosseal or
dermal delivery. The product may be provided together or separately
with one or more vitamins, minerals, amino acids, lipids, nucleic
acids, co-factors, pro-vitamins, and combinations of mixtures
thereof. Examples of odd chain fatty acids may be produced by
solution precipitation methods, anti-solvent precipitation, spray
drying, spray freezing, evaporative precipitation into an aqueous
solution, wet milling, mechanical milling, vacuum-drying,
vacuum-heating or lyophilization. The odd chain fatty acids may be
particles that are amorphous, crystalline or semi-crystalline. An
example is produced by evaporative precipitation into an aqueous
solution and the formulation has a drug-to-excipient ratio greater
than about 3:1. The odd chain fatty acids may be a provided as a
ketogenic odd-chain fatty acid-to-excipient ratio that is greater
than about 1:10, 1:1, 5:1, 7:1 or 10:1. For example, the ketogenic
odd-chain fatty acid-to-emulsifier ratio is about 1:10, 1:1, 4:1,
7:1, 10:1, 13:1, 15:1, 25:1 or 40:1, e.g., C7:Imwittor.
[0018] The present invention may also be used in a method of
providing nutritional support for surgery by providing a patient in
need of supplemental metabolic support a nutritionally effective
amount of a ketogenic odd-chain fatty acid of between about 30-35%
of total Kcal/day. The nutritional supplements may also be used in
a method for ameliorating the effects of physical exertion, the
method includes administration to a person in need of such
amelioration a pharmaceutically or nutritionally effective amount
of one or more of the odd chain fatty acids described herein. The
odd chain fatty acids will also find particular uses as a
post-operative nutritional support that includes a nutritionally
effective amount of a compound that provides both 2-carbon and
3-carbon citric acid cycle metabolites. An example of a 2-carbon
metabolite is acetyl-CoA and the 3-carbon metabolite is
propionyl-CoA. The composition may also include a nutritionally
effective amount of one or more essential fatty acids and/or one or
more essential saccharides.
[0019] The odd chain fatty acids may even be provided in a modified
release product having two portions, wherein a first portion
includes a first quantity of a ketogenic odd-chain fatty acid in an
immediate release form which becomes fully bioavailable in the
subject's stomach and a second portion with a second compound in a
sustained release form wherein the ratio of the first quantity to
the second quantity provides a serum Cmax in a human subject
equivalent to the serum Cmax obtained when the first of one or more
doses of a standard immediate release formulation having one third
the amount of the ketogenic odd-chain fatty acid is dosed every
four hours over a 12 hour period and wherein the product also
provides therapeutically effective bioavailability for at least
twelve hours after a single dose in a human subject according to
serum analysis. The second compound may even by the ketogenic
odd-chain fatty acid and/or one or more vitamins, minerals, amino
acids, lipids, nucleic acids, co-factors, pro-vitamins, and
combinations of mixtures thereof.
[0020] A diet is provided by the present invention that includes a
ketogenic odd-chain fatty acid with between about 5 to 40% of total
Kcal/day, essential fats of between about 5-20% total Kcal/day,
carbohydrates restricted to about 20 to 40% total Kcal/day of the
diet and a protein content of between about 15 to 30% total
Kcal/day. The total Kcal/day of each of the fatty acid, fats,
carbohydrates and/or protein can be varied depending on the
particular need and application. For example, the essential fats
provides between about 10-15% of the total Kcal/day, the daily
carbohydrate intake provides about 30% of the total Kcal/day, the
daily protein intake provides about 20% of the total Kcal/day or
the ketogenic odd-chain fatty acid provides between about 1-4
gm/Kg/day. The skilled artisan will recognize the different
variations and combinations available.
[0021] The ketogenic odd-chain fatty acid includes one or more
odd-chain fatty acids selected from C5, C7, C15, and mixtures or
combinations thereof. In addition, the ketogenic odd-chain fatty
acid includes triheptanoin, n-heptanoic acid, a triglyceride, or a
salt or derivative thereof, or combinations thereof.
[0022] The present invention provides a beverage including water,
carbohydrates, electrolytes and a ketogenic odd-chain fatty acid in
a concentration of between about 0.5% to about 5.0%. The beverage
includes ketogenic odd-chain fatty acid that have one or more
odd-chain fatty acids selected from C5, C7, C15, and mixtures or
combinations thereof The ketogenic odd-chain fatty acid has a
concentration of about 1.0% and in some instances, the fatty acid
is emulsified. The beverage includes ketogenic odd-chain fatty acid
that provides between about 20-30% of the daily caloric intake. The
ingredients of the beverage include the following:
TABLE-US-00001 Ingredient Approximate Concentration Potassium 2
meq/l Sodium 26 meq/l Glucose 4% Pyruvate 1% ketogenic odd-chain
fatty acid 1 to 20% Emulsifier 0.1 to 2.0% water balance.
[0023] A beverage is also provided that provides a source of
immediate energy that has a composition by weight:
TABLE-US-00002 Total Carbohydrate 0.4-3.5% Ketogenic odd-chain
fatty acid 0.1 to 20% Emulsifier 0.1 to 2.0% Sodium Chloride
0.16-0.33% Potassium Chloride 0.03-0.13% Free Citric Acid
0.026-0.26% Water balance
[0024] In addition to beverages, the present invention includes a
dry beverage concentrate for enhanced, immediate energy
requirements in a mammalian body that includes a composition by
weight in water: an emulsified, ketogenic odd-chain fatty acid
0.1-25%, total carbohydrate of 0.4-3.5%, sodium chloride
0.16-0.22%, potassium chloride 0.03-0.13%, and free citric acid
0.026-0.26%, the concentrate suitable for mixture with water to
yield a beverage. The skilled artisan will recognize that the
weight of the component may be varied depending on the particular
application.
[0025] A food is also provided that includes a mixture of
ingredients selected to make one or more snacks, soups, salads,
cakes, cookies, crackers, breads, ice creams, yogurts, puddings,
custards, baby foods, medicinal foods, sports bars, breakfast
cereals and beverages and a ketogenic odd-chain fatty acid in a
concentration of between about 0.5% to about 5.0% of the
composition. The food composition includes grains, fruits, nuts and
supplemental dietary fiber in the form of compressed flakes,
supplemental dietary fiber and combinations thereof. The present
invention provides a food composition that includes compressed
flakes of supplemental dietary fiber in the form of apple fiber,
corn bran, soy fiber, pectin, guar gum, gum ghatti, and gum arabic,
as well as mixtures thereof Some embodiments of the present
invention include one or more binder materials that include rice
flour, wheat flour, oat flour, corn flour, rye flour and potato
flour, as well as mixtures thereof
[0026] The present invention also includes a pre-cooked edible and
chewable product selected from the group consisting of breakfast
cereals, snacks, soups, salads, cakes, cookies, crackers, puddings,
ice creams, yogurts, puddings, custards, baby foods, medicinal
foods, sports bars, and beverages. Additionally, the composition
can be extrusion cooked. The present invention includes an
ingredient for fortification of a food product that has an
emulsified ketogenic odd-chain fatty acid in a concentration of
between about 0.5% to about 5.0% of the food supplement.
[0027] An orally administratable partially dry unit dosage is
provided that includes at least about 10 to 1,000 mg of a ketogenic
odd-chain fatty acid. When in the form of a nutritional supplement
includes a nutritionally effective amount of a ketogenic odd-chain
fatty acid that are metabolized into both 2-carbon and 3-carbon
citric acid cycle metabolites, and the ketogenic odd-chain fatty
acid is selected from the group consisting of a C5, C7, C15, and
mixtures or combinations thereof.
[0028] The present invention also includes a method for increasing
the immediately available energy supply to a cell by exposing the
cell to an effective dosage of a ketogenic odd-chain fatty acid
that represents 20-35% of the cellular requirement of kCals per
day.
[0029] A method for increasing the immediately available energy
supply to a mammal by providing the mammal with an effective dosage
of a ketogenic odd-chain fatty acid that comprises 30-35% of the
mammal's kCal per day is also included. The ketogenic odd-chain
fatty acid includes one or more odd-chain fatty acids selected from
C5, C7, C15, and mixtures or combinations thereof.
[0030] The present invention also includes an animal feed that has
an odd chain fatty acid present in an amount effective to provide
nutritive fat to an animal and a solid nutritive source. The animal
feed is adapted for use by one or more of poultry, livestock,
farm-raised fish, crabs, shrimp and fresh-water turtles. The solid
nutrititive source includes a source selected from the group
consisting of soy, oats, sorghum, whole wheat, a nutritive wheat
fraction, whole rice, a nutritive rice fraction, whole corn a
nutritive corn fraction and whole barley, a nutritive barley
fraction, vitamins and nutritive minerals. When used as an animal
feed the odd chain fatty acid is present in an amount of at least
about 5% to 40% by weight of the feed.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] For a more complete understanding of the features and
advantages of the present invention, reference is now made to the
detailed description of the invention along with the accompanying
figures and in which:
[0032] FIG. 1 is a schematic drawing depicting the metabolic fate
of the odd-carbon fatty acid, heptanoate (C7), derived from the
triglyceride, Triheptanoin;
[0033] FIG. 2 is a schematic drawing depicting the organellar
integration of glycogen metabolism in cells;
[0034] FIG. 3 is a schematic drawing depicting the possible enzyme
deficiencies in inherited defects involving glycogen metabolism in
humans. (GSD I-GSD VII);
[0035] FIG. 4 is a schematic drawing depicting the effect of
odd-carbon therapy on glycogen metabolic disorders, including GSD
II, in humans;
[0036] FIG. 5 depicts the effect of a triheptanoin-containing diet
on the levels of selected amino acids in the blood of a patient
with the adult onset form of glycogen storage disease II (acid
maltase deficiency) with a diet containing Triheptanoin at a dose
of 1.0 to 1.5 gm/kg/day);
[0037] FIG. 6 is a diagram that shows the oxidative sequence for
heptanoate (liver);
[0038] FIG. 7 is a diagram that shows the entry points in
metabolism, e.g., in the CAC of the odd chain fatty acids of the
present invention; and
[0039] FIG. 8 is a graph that compared the acylcarnitine levels
after trihepatoin treatment in horses; and
[0040] FIG. 9 is a graph that shows the effects of a resolution of
rhabdomyolysis with dietary triheptanoin in various types of
FODs.
DETAILED DESCRIPTION OF THE INVENTION
[0041] While the making and using of various embodiments of the
present invention are discussed in detail below, it should be
appreciated that the present invention provides many applicable
inventive concepts that can be embodied in a wide variety of
specific contexts. The specific embodiments discussed herein are
merely illustrative of specific ways to make and use the invention
and do not delimit the scope of the invention.
[0042] To facilitate the understanding of this invention, a number
of terms are defined below. Terms defined herein have meanings as
commonly understood by a person of ordinary skill in the areas
relevant to the present invention. Terms such as "a", "an" and
"the" are not intended to refer to only a singular entity, but
include the general class of which a specific example may be used
for illustration. As such, unless otherwise indicated, the terms
"a" and "an" are taken to mean "one", "at least one" or "one or
more". The terminology herein is used to describe specific
embodiments of the invention, but their usage does not delimit the
invention, except as outlined in the claims.
[0043] As used herein, the terms "subject" or "patient" are
intended to include living organisms that may have one or more
conditions generally referred to as polysaccharide storage
diseases. Examples of subjects include humans, monkeys, horses,
cows, sheep, goats, dogs, cats, mice, rats, and transgenic species
thereof. Other examples of subjects include experimental animals
such as mice, rats, dogs, cats, goats, sheep, pigs, and cows. A
subject can be a human suffering from, or suspected of having, a
polysaccharide storage disease, e.g., a glycogen storage disease
such as Pompe's disease (GSD II).
[0044] As used herein, the phrases "therapeutically effective
dosage" or "therapeutically effective amount" is an amount of a
compound or mixtures of compounds, such as the odd-chain fatty
acids and precursors or derivatives thereof, that reduce the amount
of one or more symptoms of the condition in the infected subject by
at least about 20%, at least about 40%, even more at least about
60%, 80% or even 100% relative to untreated subjects. Active
compounds are administered at a therapeutically effective dosage
sufficient to treat a condition associated with a condition in a
subject. For example, the efficacy of a compound can be evaluated
in patients or animal model systems that may be predictive of
efficacy in treating the disease in humans or animals.
[0045] As used herein the term, "essential fatty acids" is used to
describe fats and oils in foods are made up of basic units called
fatty acids. In the body, these typically travel in three's as
fatty acid chains attached to glycerol, forming a triglyceride.
Based on their chemical structure, fatty acids are classified into
3 major categories: monounsaturated, polyunsaturated, or saturated
fats. The oils and fats that people and animals eat are nearly
always mixtures of these 3 types of fatty acids, with one type
predominating. Two specific types of polyunsaturated fatty acids,
linoleic and alpha-linolenic, are called essential fatty acids.
They must be present in the diet in adequate amounts because they
are considered necessary for proper nutrition and health. Linoleic
acid (LA) is an omeaga-6 fatty acid and is found in many oils,
e.g., corn, safflower, soybean and sunflower, whole grains and
walnuts. Alpha-linolenic acid (ALA) is a plant precursor of
docosahexanoic acid (DHA). Sources of ALA include seaweeds and
green leaves of plants (in very small amounts), soybeans, walnuts,
butternuts, some seeds (flax, chia, hemp, canola) and the oils
extracted from these foods.
[0046] As used herein, the term "nutritionally effective amount" is
used to mean the amount of odd chain fatty acids that will provide
a beneficial nutritional effect or response in a mammal. For
example, as with a nutritional response to vitamin- and
mineral-containing dietary supplements varies from mammal to
mammal, it should be understood that nutritionally effective
amounts of the odd chain fatty acids will vary. Thus, while one
mammal may require a particular profile of vitamins and minerals
present in defined amounts, another mammal may require the same
particular profile of vitamins and minerals present in different
defined amounts. Such is the case with the nutritionally effective
amounts of the odd chain fatty acids of the present invention, in
which the supplementation may be used to add C3 and C2 carbon
chains into the liver and/or the heart, muscle, brain and
kidney.
[0047] When provided as a dietary supplement or additive, the odd
chain fatty acids of the invention has been prepared and
administered to mammals in powdered, reconstitutable powder,
liquid-solid suspension, liquid, capsule, tablet, caplet, lotion
and cream dosage forms. The skilled artisan in the science of
formulations can use the odd chain fatty acids disclosed herein as
a dietary supplement that may be formulated appropriately for,
e.g., irrigation, ophthalmic, otic, rectal, sublingual,
transdermal, buccal, vaginal, or dermal administration. Thus, other
dosage forms such as chewable candy bar, concentrate, drops,
elixir, emulsion, film, gel, granule, chewing gum, jelly, oil,
paste, pastille, pellet, shampoo, rinse, soap, sponge, suppository,
swab, syrup, chewable gelatin form, chewable tablet and the like,
can be used.
[0048] Due to varying diets among people, the dietary odd chain
fatty acids of the invention may be administered in a wide range of
dosages and formulated in a wide range of dosage unit strengths. It
should be noted that the dosage of the dietary supplement can also
vary according to a particular ailment or disorder that a mammal is
suffering from when taking the supplement. For example, a person
suffering from chronic fatigue syndrome or fibromyalgia will
generally require a dose different than an athlete that is wanting
to attain a nutritional benefit. An appropriate dose of the dietary
supplement can be readily determined by monitoring patient
response, i.e., general health, to particular doses of the
supplement. The appropriate doses of the supplement and each of the
agents can be readily determined in a like fashion by monitoring
patient response, i.e., general health to particular doses of
each.
[0049] The odd chain fatty acids may be administered simultaneously
or sequentially in one or a combination of dosage forms. While it
is possible and even likely that the present dietary supplement
will provide an immediate overall health benefit, such benefit may
take days, weeks or months to materialize. Nonetheless, the present
dietary odd chain fatty acid supplement will provide a beneficial
nutritional response in a mammal consuming it.
[0050] The odd-chain fatty acids of the present invention may be
administered, e.g., orally or by subcutaneous, intravenous,
intraperitoneal, etc., administration (e.g. by injection).
Depending on the route of administration, the active compound may
be neutralized, made miscible, at least partially or fully
water-soluble or even coated in a material to protect the odd-chain
fatty acids from the action of bases, acids, enzymes or other
natural conditions that may interfere with their effectiveness,
uptake or metabolic use.
[0051] To administer the therapeutic compound by other than
parenteral administration, it may be necessary to coat the compound
with, or co-administer the compound with, a material to prevent its
inactivation. For example, the therapeutic compound may be
administered to a subject in an appropriate carrier, for example,
emulsifiers, liposomes, or a diluent. Pharmaceutically acceptable
diluents include saline and aqueous buffer solutions. The
therapeutic odd-chain fatty acids may be dispersed in glycerol,
liquid polyethylene glycols, and mixtures thereof and in oils.
Under ordinary conditions of storage and use, these preparations
may contain a preservative to prevent the growth of
microorganisms.
[0052] Pharmaceutical compositions that include the odd-chain fatty
acids of the present invention suitable for injectable use may
include sterile aqueous solutions, dispersions and sterile powders
for the extemporaneous preparation of sterile injectable solutions
or dispersion. In all cases, the composition must be sterile and
must be fluid to the extent that easy syringability exists. It must
be stable under the conditions of manufacture and storage and must
be preserved against the contaminating action of microorganisms
such as bacteria and fungi.
[0053] The odd-chain fatty acids may be provided with a carrier in
a solvent or dispersion medium containing, for example, water,
ethanol, polyol (for example, glycerol, propylene glycol, and
liquid polyethylene glycol, and the like), suitable mixtures
thereof, and vegetable oils. The proper fluidity can be maintained,
for example, by the use of a coating such as lecithin, by the
maintenance of the required particle size in the case of dispersion
and by the use of surfactants. Prevention of the action of
microorganisms can be achieved by various antibacterial and
antifungal agents, for example, parabens, chlorobutanol, phenol,
ascorbic acid, thimerosal, and the like. In many cases, it will be
preferable to include isotonic agents, for example, sugars, sodium
chloride, or polyalcohols such as mannitol and sorbitol, in the
composition. Prolonged absorption of the injectable compositions
can be brought about by including in the composition an agent which
delays absorption, for example, aluminum monostearate or
gelatin.
[0054] The odd-chain fatty acids may be provided in one or more
controlled sizes and characteristics with one or more water-soluble
polymers depending on the size and structural requirements of the
patient, e.g., the particles may be small enough to traverse blood
vessels when provided intravenously. Either synthetic or naturally
occurring polymers may be used, and while not limited to this
group, some types of polymers that might be used are
polysaccharides (e.g. dextran, ficoll), proteins (e.g.
poly-lysine), poly(ethylene glycol), or poly(methacrylates).
Different polymers, because of their different size and shape, will
produce different diffusion characteristics for the odd-chain fatty
acids in the target tissue or organ.
[0055] Sterile injectable solutions can be prepared by
incorporating the therapeutic compound in the required amount in an
appropriate solvent with one or a combination of ingredients
enumerated above, as required, followed by filtered sterilization.
Generally, dispersions are prepared by incorporating the
therapeutic compound into a sterile carrier which contains a basic
dispersion medium and the required other ingredients from those
enumerated above. In the case of sterile powders for the
preparation of sterile injectable solutions, methods of preparation
include: vacuum drying, spray freezing, freeze-drying and the like,
which yield a powder of the active ingredient (i.e., the
therapeutic compound) plus any additional desired ingredient from a
previously sterile-filtered solution thereof.
[0056] The odd-chain fatty acids can be orally administered, for
example, with an inert diluent or an assimilable edible carrier.
The therapeutic compound and other ingredients may also be enclosed
in a hard or soft shell gelatin capsule, compressed into tablets,
or incorporated directly into the subject's diet. The odd-chain
fatty acids may be incorporated with excipients and used in the
form of ingestible tablets, buccal tablets, troches, capsules,
elixirs, suspensions, syrups, wafers, and the like. The amount of
odd-chain fatty acids in the compositions and preparations may, of
course, be varied depending on, e.g., the age, weight, gender,
condition, disease and course of treatment of the individual
patient. Pediatric doses are likely to differ from adult doses as
will be known to the skilled artisan. The amount of the therapeutic
compound in such therapeutically useful compositions is such that a
suitable dosage will be obtained.
[0057] A dosage unit for use with the odd chain fatty acids
disclosed herein may be a single compound or mixtures thereof with
other compounds, e.g., amino acids, nucleic acids, vitamins,
minerals, pro-vitamins and the like. The compounds may be mixed
together, form ionic or even covalent bonds. For pharmaceutical
purposes the odd chain fatty acids (e.g., C5, C7 and C15) of the
present invention may be administered in oral, intravenous (bolus
or infusion), intraperitoneal, subcutaneous, or intramuscular form,
all using dosage forms well known to those of ordinary skill in the
pharmaceutical arts. Depending on the particular location or method
of delivery, different dosage forms, e.g., tablets, capsules,
pills, powders, granules, elixirs, tinctures, suspensions, syrups,
and emulsions may be used to provide the odd chain fatty acids of
the present invention to a patient in need of therapy that includes
a number of conditions, e.g., polysaccharide storage diseases,
fatigue, low energy, wasting and the like. The odd chain fatty
acids may also be administered as any one of known salt forms.
[0058] The total daily amount of odd chain fatty acids will vary
depending on the condition and needs of a patient. For example, the
odd chain fatty acids may be provided as a supplemental source of
immediate, short-term, mid-term or long-term energy and may be
provided in formulations that are immediately available, slow
release or extended release. The dosage amount may be measured in
grams per day, as a percentage of kCalories consumed in a day, as a
percentage of the total daily caloric intake, as part of a fixed, a
modified or a diet that changes over time. For example, a patient
may need immediate intervention that "spikes" the amount of odd
chain fatty acids to approach or reach ketosis. These "ketogenic"
odd chain fatty acids will then be varied to not have other side
effects, e.g., start with 40% of total caloric intake per day and
then reduced over time as the patient's condition, symptoms,
clinical course and/or metabolic conditions improves. The range of
percentage caloric intake may vary from between about 0.01, 0.1, 1,
2, 5, 10, 15, 20, 22, 25, 30, 35, 40 or even higher percent, which
may include one or more of the odd chain fatty acids (e.g., C5, C7
or C15 (available from, e.g., Sassol, Germany). One way to measure
the effect and/or dosing of the odd chain fatty acids is to measure
the amount that is detectable in body solids or fluids, e.g.,
biopsies and blood, respectively. A wide variety of odd chain fatty
acids metabolites may be detected from multiple sources, e.g.,
urine, tears, feces, blood, sweat, breath and the like.
[0059] For example, when using C7 as the source of odd chain fatty
acids these can be provided in the form of a triglyceride, e.g.,
tri-heptanoin. The triglyceride triheptanoin is provided in a
concentration sufficient to provide a beneficial effect is most
useful in this aspect of the present invention. The seven-carbon
fatty acid may be provided, e.g.:
TABLE-US-00003 Infants 1-4 g/kg 35% kcalories Children 3-4 g/kg
33-37% kcalories Adolescent 1-2 g/kg 35% kcalories Adults 0.1-2
g/kg 35% kcalories
[0060] Goals have been set using 4 g/kg (within ideal body weight
(IBW) range) for infants, children, and some adolescents. Goals
have been set using 2 g/kg (within IBW range) for adolescents.
Goals have been set using 2 g/kg (within IBW range) for adults; but
toleration is 1-1.2 g per kg (which is 35% kcal of estimated
needs).
[0061] The odd chain fatty acids are typically administered in
admixture with suitable pharmaceutical salts, buffers, diluents,
extenders, excipients and/or carriers (collectively referred to
herein as a pharmaceutically acceptable carrier or carrier
materials) selected based on the intended form of administration
and as consistent with conventional pharmaceutical practices.
Depending on the best location for administration, the odd chain
fatty acids may be formulated to provide, e.g., maximum and/or
consistent dosing for the particular form for oral, rectal,
topical, intravenous injection or parenteral administration. While
the odd chain fatty acids may be administered alone or pure, they
may also be provided as stable salt form mixed with a
pharmaceutically acceptable carrier. The carrier may be solid or
liquid, depending on the type and/or location of administration
selected.
[0062] Techniques and compositions for making useful dosage forms
using the present invention are described in one or more of the
following references: Ansel, Introduction to Pharmaceutical Dosage
Forms 2nd Edition (1976); Remington's Pharmaceutical Sciences, 17th
ed. (Mack Publishing Company, Easton, Pa., 1985); Advances in
Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds.,
1992); Advances in Pharmaceutical Sciences Vol 7. (David Ganderton,
Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric
Coatings for Pharmaceutical Dosage Forms (Drugs and the
Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989);
[0063] Pharmaceutical Particulate Carriers: Therapeutic
Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain
Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract
(Ellis Horwood Books in the Biological Sciences. Series in
Pharmaceutical Technology; J. G. Hardy, S. S. Davis, Clive G.
Wilson, Eds.); Modern Pharmaceutics Drugs and the Pharmaceutical
Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.),
and the like, relevant portions of each incorporated herein by
reference.
[0064] Odd chain fatty acids may be administered in the form of an
emulsion and/or liposome, e.g., small unilamellar vesicles, large
unilamallar vesicles and multilamellar vesicles, whether charged or
uncharged. Liposomes may include one or more: phospholipids (e.g.,
cholesterol), stearylamine and/or phosphatidylcholines, mixtures
thereof, and the like. Examples of emulsifiers for use with the
present invention include: Imwitor 370, Imwitor 375, Imwitor 377,
Imwitor 380 and Imwitor 829.
[0065] The odd chain fatty acid vesicles may also be coupled to one
or more soluble, biodegradable, bioacceptable polymers as drug
carriers or as a prodrug. Such polymers may include:
polyvinylpyrrolidone, pyran copolymer,
polyhydroxylpropylmethacrylamide-phenol,
polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysine
substituted with palmitoyl residues, mixtures thereof, and the
like. Furthermore, the vesicles may be coupled one or more
biodegradable polymers to achieve controlled release of the odd
chain fatty acids. Biodegradable polymers for use with the present
invention include, e.g., polylactic acid, polyglycolic acid,
copolymers of polylactic and polyglycolic acid, polyepsilon
caprolactone, polyhydroxy butyric acid, polyorthoesters,
polyacetals, polydihydropyrans, polycyanoacylates, and crosslinked
or amphipathic block copolymers of hydrogels, mixtures thereof, and
the like.
[0066] In one embodiment, gelatin capsules (gelcaps) may include
the odd chain fatty acid in its native state. For oral
administration in a liquid dosage form, the oral drug components
may be combined with any oral, non-toxic, pharmaceutically
acceptable inert carrier such as an emulsifier, a diluent or
solvent (e.g., ethanol), glycerol, water, and the like. Examples of
suitable liquid dosage forms include oily solutions or suspensions
in water, pharmaceutically acceptable fats and oils, alcohols or
other organic solvents, including esters, emulsions, syrups or
elixirs, suspensions, solutions and/or suspensions reconstituted
from non-effervescent granules and even effervescent preparations
reconstituted from effervescent granules. Such liquid dosage forms
may contain, for example, suitable solvents, preservatives,
emulsifying agents, suspending agents, diluents, sweeteners,
thickeners, and melting agents, mixtures thereof, and the like.
[0067] Liquid dosage forms for oral administration may also include
coloring and flavoring agents that increase patient acceptance and
therefore compliance with a dosing regimen. In general, water, a
suitable oil, saline, aqueous dextrose (e.g., glucose, lactose and
related sugar solutions) and glycols (e.g., propylene glycol or
polyethylene glycols) may be used as suitable carriers for
parenteral solutions. Solutions for parenteral administration
include generally, a water soluble salt of the active ingredient,
suitable stabilizing agents, and if necessary, buffering salts.
Antioxidizing agents such as sodium bisulfite, sodium sulfite
and/or ascorbic acid, either alone or in combination, are suitable
stabilizing agents. Citric acid and its salts and sodium EDTA may
also be included to increase stability. In addition, parenteral
solutions may include pharmaceutically acceptable preservatives,
e.g., benzalkonium chloride, methyl- or propyl-paraben, and/or
chlorobutanol. Suitable pharmaceutical carriers are described in
multiple editions of Remington's Pharmaceutical Sciences, Mack
Publishing Company, a standard reference text in this field,
relevant portions incorporated herein by reference.
[0068] For direct delivery to the nasal passages, sinuses, mouth,
throat, esophagus, trachea, lungs and alveoli, the odd chain fatty
acids may also be delivered as an intranasal form via use of a
suitable intranasal vehicle. For dermal and transdermal delivery,
the odd chain fatty acids may be delivered using lotions, creams,
oils, elixirs, serums, transdermal skin patches and the like, as
are well known to those of ordinary skill in that art. Parenteral
and intravenous forms may also include pharmaceutically acceptable
salts and/or minerals and other materials to make them compatible
with the type of injection or delivery system chosen, e.g., a
buffered, isotonic solution.
[0069] To the extent that the odd chain fatty acids may be made
into a dry powder or form, they may be included in a tablet.
Tablets will generally include, e.g., suitable binders, lubricants,
disintegrating agents, coloring agents, flavoring agents,
flow-inducing agents and/or melting agents. For example, oral
administration may be in a dosage unit form of a tablet, gelcap,
caplet or capsule, the active drug component being combined with a
non-toxic, pharmaceutically acceptable, inert carrier such as
lactose, gelatin, agar, starch, sucrose, glucose, methyl cellulose,
magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol,
sorbitol, mixtures thereof, and the like. Suitable binders for use
with the present invention include: starch, gelatin, natural sugars
(e.g., glucose or beta-lactose), corn sweeteners, natural and
synthetic gums (e.g., acacia, tragacanth or sodium alginate),
carboxymethylcellulose, polyethylene glycol, waxes, and the like.
Lubricants for use with the invention may include: sodium oleate,
sodium stearate, magnesium stearate, sodium benzoate, sodium
acetate, sodium chloride, mixtures thereof, and the like.
Disintegrators may include: starch, methyl cellulose, agar,
bentonite, xanthan gum, mixtures thereof, and the like.
[0070] Capsules. Capsules may be prepared by filling standard
two-piece hard gelatin capsules each with 10 to 500 milligrams of
powdered active ingredient, 5 to 150 milligrams of lactose, 5 to 50
milligrams of cellulose and 6 milligrams magnesium stearate.
[0071] Soft Gelatin Capsules. The odd chain fatty acids may be
dissolved in an oil, e.g., a digestible oil such as soybean oil,
cottonseed oil or olive oil. Non-digestible oils may also be used
to have better control over the total caloric intake provided by
the oil. The active ingredient is prepared and injected by using a
positive displacement pump into gelatin to form soft gelatin
capsules containing, e.g., 100-500 milligrams of the active
ingredient. The capsules are washed and dried.
[0072] Tablets. A large number of tablets are prepared by
conventional procedures so that the dosage unit was 100-500
milligrams of active ingredient, 0.2 milligrams of colloidal
silicon dioxide, 5 milligrams of magnesium stearate, 50-275
milligrams of microcrystalline cellulose, 11 milligrams of starch
and 98.8 milligrams of lactose. Appropriate coatings may be applied
to increase palatability or delay absorption.
[0073] To provide an effervescent tablet, appropriate amounts of,
e.g., monosodium citrate and sodium bicarbonate, are blended
together and then roller compacted, in the absence of water, to
form flakes that are then crushed to give granulates. The
granulates are then combined with the active ingredient, drug
and/or salt thereof, conventional beading or filling agents and,
optionally, sweeteners, flavors and lubricants.
[0074] Injectable solution. A parenteral composition suitable for
administration by injection is prepared by stirring sufficient
active ingredient in deionized water and mixed with, e.g., up to
10% by volume propylene glycol, salts and/or water to deliver a
composition, whether in concentrated or ready-to-use form. Given
the nature of the odd chain fatty acids (alone, partially or
fully-soluble in water) the amount and final concentration of the
odd chain fatty acids may be varied such that the liquid may be
provided intravenously using syringes and/or standard intravenous
liquids or fluids. The solution will generally be made isotonic
with sodium chloride and sterilized using, e.g.,
ultrafiltration.
[0075] Suspension. An aqueous suspension is prepared for oral
administration so that each 5 ml contain 100 mg of finely divided
active ingredient, 200 mg of sodium carboxymethyl cellulose, 5 mg
of sodium benzoate, 1.0 g of sorbitol solution, U.S.P., and 0.025
ml of vanillin.
[0076] Mini-tablets. For mini-tablets, the active ingredient is
compressed into a hardness in the range 6 to 12 Kp. The hardness of
the final tablets is influenced by the linear roller compaction
strength used in preparing the granulates, which are influenced by
the particle size of, e.g., the monosodium hydrogen carbonate and
sodium hydrogen carbonate. For smaller particle sizes, a linear
roller compaction strength of about 15 to 20 KN/cm may be used.
[0077] Kits. The present invention also includes pharmaceutical
kits useful, for example, for providing an immediate source of
alternative cellular energy, e.g., before, during or after surgery.
The dosage will generally be prepared sterile and ready-to-use,
e.g., one or more containers that may be broken (e.g., sealed glass
ampoules), pierced with a syringe for immediate administration or
even a pressurized container. Such kits may further include, if
desired, one or more of various conventional pharmaceutical kit
components, such as, for example, containers with one or more
pharmaceutically acceptable diluents, carriers, additional
containers, etc., as will be readily apparent to those skilled in
the art. Printed instructions, either as inserts or as labels,
indicating quantities of the components to be administered,
guidelines for administration, and/or guidelines for mixing the
components, may also be included in the kit. It should be
understood that although the specified materials and conditions are
important in practicing the invention, unspecified materials and
conditions are not excluded so long as they do not prevent the
benefits of the invention from being realized.
[0078] Pharmaceutical Dosage Forms. The odd chain fatty acids of
the present invention may be provided in liquid form or may also be
provided in a capsule, gelcap or other encapsulated form.
[0079] Generally, one composition of the present invention is
prepared by adding, e.g., half of the Kaolin clay or other carrier
into the blended followed by addition of a first active salt form,
e.g., the salt form that is less soluble in the final liquid
suspension, e.g., as an emulsion in water. This process is
particularly suitable for very large mixtures, e.g., 500, 1,000,
3,000 or even 5,000 liters.
[0080] One particular method of delivery of the odd chain fatty
acids of the present invention is in a tablet, capsule or gelcap
that is coated for enteric delivery. Enteric coating relates to a
mixture of pharmaceutically acceptable excipients that is applied
to, combined with, mixed with or otherwise added to a carrier to
deliver the medicinal content, in this case one or more odd chain
fatty acids (e.g., C5, C7, C15, mixtures and combinations thereof)
through the stomach unaltered for delivery into the intestines. The
coating may be applied to a compressed or molded or extruded
tablet, a gelatin capsule, and/or pellets, beads, granules or
particles of the carrier or composition. The coating may be applied
through an aqueous dispersion or after dissolving in appropriate
solvent. Additional additives and their levels, and selection of a
primary coating material or materials will depend on the following
properties: resistance to dissolution and disintegration in the
stomach; impermeability to gastric fluids and drug/carrier/enzyme
while in the stomach; ability to dissolve or disintegrate rapidly
at the target intestine site; physical and chemical stability
during storage; non-toxicity; easy application as a coating
(substrate friendly); and economical practicality. Methods for
enteric coating are well known in the art.
[0081] Remington's Pharmaceutical Sciences, discloses that enteric
polymer carries generally include carboxyl groups and hydrophobic
groups in the molecule and the enteric polymer is dissolved in a
solvent having a specific pH value through the dissociation of the
carboxyl groups. For instance, commercially available
hydroxypropylmethyl cellulose acetate succinate is a derivative of
hydroxypropylmethyl cellulose which is substituted with carboxyl
groups (succinoyl groups) and hydrophobic groups (acetyl groups).
Alginic acid, sodium alginate other natural materials may also be
used to provide an enteric coating.
[0082] Other additives and excipients may then be added to the
formulation of the partially water soluble carrier-active odd chain
fatty acids mixture, e.g., adding Povidone (e.g., Povidone 30),
Xantham gum (or other gums) and Sorbitol to a mixture of Kaolin
Clay to provide a specific example of one formulation of the
present invention. As will be apparent to those of skill in the
art, the actual amount of the partially-excipient soluble active
salt (e.g., non or partially water soluble) may be varied in
accordance with the dissolution characteristics of the active,
which may be further varied by addition of agents that affect the
solubility and/or dissolution of the active in, e.g., water. As
regards a pediatric formulation, the amount of active may be
reduced in accordance with the dosage form approved for pediatric
use.
[0083] One example of a liquid odd chain fatty acid(s)
pharmaceutical composition may be prepared with the following
components:
TABLE-US-00004 Components Weight Odd chain fatty acid(s) 1.0 Kg
emulsifier (e.g., Imwitor 375) 100 gr Purified water (USP) 2.0
Kg
The formulation may further include, e.g.:
TABLE-US-00005 Glycerin (USP) 500.0 ml Sorbitol Solution, 70% (USP)
500.0 ml Saccharin Sodium (USP) 10.0 gr Citric Acid (USP) 10.0 gr
Sodium Benzoate (NF) 6.0 gr Kollidon 30 330.0 gr Xanthan Gum 200
Mesh 20.0 gr Bubble Gum Flavor 11.1 gr Methylparaben 1.0 gr
Proplyparaben 100 mg Propylene Glycol (USP) 75 ml Additional
ddH.sub.2O QS to 5 liters.
With appropriate increases of the above for scale-up.
[0084] A batch of mixed release odd chain fatty acids in an
enveloped preparation on a carrier, e.g., beads, may be prepared
with the following components:
TABLE-US-00006 Components Weight Emulsified odd chain fatty acids
8.0 mg Carrier 51.7 mg Calcium Stearate 4.0 mg Talc 4.0 mg
Pharmaceutical Glaze 5.5 mg
[0085] When combining odd chain fatty acids (C5, C7 and/or C15),
these may be formulated as follows. A capsule for extended release
of a first active and extended release of a second active in an
enveloped formulation, in a single capsule:
TABLE-US-00007 First Bead Weight Second Bead Weight odd chain fatty
acid 6.0 mg odd chain fatty acid C15 2.0 mg C7 Bead 162.9 mg Bead
108.5 mg Lacquer 6 mg Lacquer 3.3 mg Talc 12.6 mg Talc 5 mg Calcium
Stearate 12.6 mg Calcium Stearate 5 mg Capsule 1
[0086] When combining the odd chain fatty acids, these may be
formulated as follows. A capsule for extended release of a first
active and extended release of a second active in an enveloped
formulation, in a single capsule:
TABLE-US-00008 First Bead Weight Second Bead Weight odd chain fatty
acid 6.0 mg odd chain fatty acids C7 2.0 mg C5 Bead 162.9 mg Bead
108.5 mg Lacquer 6 mg Lacquer 3.3 mg Talc 12.6 mg Talc 5 mg Calcium
Stearate 12.6 mg Calcium Stearate 5 mg Mini-capsule 1
[0087] A formulation for extended release of odd chain fatty acids
of a second active in an enveloped formulation, in a gelcap:
TABLE-US-00009 Component Weight Component Weight odd chain fatty
acid 6.0 mg odd chain fatty acid 2.0 mg Bead 162.9 mg Bead 108.5 mg
Lacquer 6 mg Lacquer 3.3 mg Talc 12.6 mg Talc 5 mg Calcium Stearate
12.6 mg Calcium Stearate 5 mg Gelcap 1
[0088] A formulation for rectal release of odd chain fatty acids in
a suppository:
TABLE-US-00010 Component Weight Odd chain fatty acids 100 mg
Carrier 10 mg Talc 12.6 mg Calcium Stearate 12.6 mg
beeswax/glycerol 1-2 gr
[0089] An enteric-coated soft gelatin capsule that includes the odd
chain fatty acids (with or without an emulsifier) is made by
coating the odd chain fatty acids with a lipophilic material to
obtain granules, mixing the granules obtained in step with an oily
matrix, antioxidants and preservatives to form a lipid suspension,
mixing the lipid suspension within a soft gelatin film, and coating
the soft gelatin film to obtain an enteric coated soft gelatin
capsule.
[0090] The odd chain fatty acid(s), stearic acid and
triethanolamine are heated and mixed to form an emulsified fluid.
The resulting emulsified fluid is mixed well by a homogenizer to
obtain an emulsified suspension and enterically coated. Examples of
formulations include:
TABLE-US-00011 Component Weight Odd Chain Fatty Acids 360.0 g
Stearic acid 78.6 g Ethanolamine 21.4 g Odd Chain Fatty Acids 360.0
g Stearic acid 30.0 g Triethanolamine 20.0 g Odd Chain Fatty Acids
400.0 g Stearic acid 77.0 g Ethanolamine 23.0 g Cetyl alcohol 50.0
g Odd Chain Fatty Acids 245.0 g Stearic acid 38.5 g Ethanolamine
11.5 g Cetyl alcohol 50.0 g Carboxymethyl cellulose 25.0 g
TABLE-US-00012 TABLE 1 Recommended Daily Nutrient Intake Ranges
RECOMMENDED DAILY NUTRIENT INTAKE RANGES WITH FOD DEFECT NUTRIENT
AGE Protein Energy Fluid C 7 % of energy kcal/kg/day mL/kg % Kcal/d
INFANTS 0-<3 mo 10-12% 120 150-125 35% 3-6 mo 10-12% 115 160-130
35% 6-9 mo 10-12% 110 145-125 35% 9-12 mo 10-12% 105 135-120 35%
g/kg kcal/kg/day mL/day % Kal/d Children 1-3 years 2-2.8 102
900-1800 35% 4-6 years 2 90 1300-2300 35% 7-10 years 1.5 70
1650-3300 35% WOMEN 11-14 years 1 47 1500-3000 35% 15-18 years 0.8
40 2100-3000 35% >18 years 0.8 20-25 1400-2500 35% MEN 11-14
years 1 55 2000-3700 35% 15-18 years 0.9 45 2100-3900 35% >18
years 0.8 20-25 2000-3300 35% If patient is >20% ideal body
weight (IBW), use upper range IBW to calculate needs
[0091] It has now been found that administration of odd carbon
fatty acids that are converted or metabolized to the 5 carbon
ketone bodies 3-hydroxypentanoate (BHP) and 3-ketopentanoate (BKP)
is an effective treatment for GSDII in humans and polysaccharide
storage diseases in horses and other animals. In addition,
administration of the 5 carbon ketone bodies BHP and/or BKP in the
form of either free ketone bodies or in other forms capable of
providing the free ketones in vivo after administration is also an
effective treatment for these diseases. Such forms of BHP and/or
BKP as a triglyceride form, a polymeric form or a salt form could
be used. The method of the invention spares the patient's own
muscle protein because the patient no longer requires amino acids
from the muscle for an energy source. In addition, the patient no
longer requires the glycogen stored in the lysosomes as an energy
source. The glycogen stored in the lysosomes will gradually
decrease, because there is an intact glycogen degradative pathway
in the cytosol that can compensate for the defective lysosomal
pathway due to the absence of acid maltase (alpha glucosidase) in
GSD II. Lysosomal "acid maltase" also has debrancher enzyme
activity as well as alpha glucosidase activity. In the cytosolic
glycogenoses, glycogen can enter the lysosome and degradation can
be carried out by acid maltase since it has both glucosidase and
debrancher activities.
[0092] The effective therapeutic agent in vivo is a product of the
metabolism of C15, C7 or C5, or any other odd-carbon fatty acid
that breaks down metabolically to BHP and BKP in the subject
animal. FIG. 1 depicts the metabolic fate of C7 in humans. In
addition, the C5 ketone bodies BHP and BKP, as stated above, could
also be used directly for therapy. The degradation of C15, C7,
& C5 all lead to the production of BHP and BKP and their export
to other organ systems from the liver, as shown in FIG. 1.
[0093] Referring now to FIG. 2, the glycogen molecule is a polymer
produced from the union of glucose molecules in a straight chain of
glucose molecules joined together in a 1,4 linkage as well as
branching of glucose residues in an .alpha.1,6 linkage between
glucose residues. This structure is represented schematically by a
chain having a branch in this figure involving glucose residues
depicted in red, blue and green. The branch (.alpha.1,6 linkage) is
depicted in (red). In the cytosol, a phosphorylase enzyme is
required to remove the blue residues. A debrancher enzyme is
required to remove and relocate the green glucose residues to the
end of the .alpha.1,4 chain. The debrancher enzyme also clips the 1
to 6 branched residue as glucose-1-phosphate and the final product
is a straight chain of glucose residues (blue) in .alpha.1,4
linkage. This chain is completely degraded to glucose-1-phosphate
residues. Table 2 shows the profound effect of C7 therapy on amino
acid levels in the blood. After 13 and 41 hours the levels of
Alanine, for example, had greatly increased. The patient was then
off the C7 diet during a gastrostomy and the level decreased. Once
the C7 diet was re-initiated, the Alanine levels rapidly increased.
FIG. 5 shows the metabolism of C7 in the cytosol and the lumen of
the mitochondria and demonstrates the locations and entry of the
ketogenic odd chain fatty acids of the present invention.
TABLE-US-00013 TABLE 2 Plasma Amino Acids (uM/L) During Initiation
of Anaplerotic Therapy, Adult-Onset Glycogen Storage Disease II
(Pompe's). Event: Admit C7 = 1.0 gm/Kg NPO* C7 = 1.5 gm/Kg Time:
Baseline 13 hrs 41 hrs 65 hrs 84 hrs 108 hrs 132 hrs ALANINE
(162-572) 129 551 450 189 184 307 277 GLUTAMINE (424-720) 430 827
424 313 483 584 519 LEUCINE (60-204) 104 154 266 158 108 238 220
VALINE (108-295) 188 333 384 200 183 304 269 ISOLEUCINE (39-119) 58
81 138 87 61 144 128 *NPO for gastrostomy - IV glucose only;
**Essential amino acids
[0094] In the lysosome, another pathway exists for glycogen
degradation. The lysosomal .alpha.-glucosidase enzyme (acid
maltase) is not only a glucosidase, but also has the capability to
act as a debrancher enzyme as well. These characteristics of the
lysosomal .alpha.-glucosidase permit complete degradation of
lysosomal glycogen. These features are illustrated schematically in
FIG. 2.
[0095] Now referring to FIG. 3, the (red) bars show the location of
the various inherited deficiencies causing Glycogen Storage Disease
in humans. If any of these defects are present, it affects the
ability of the subject to receive adequate energy from stored
glycogen or polysaccharide. See: Chen, Y T: Glycogen Storage
Diseases. In: The Metabolic and Molecular Bases of Inherited
Diseases, 8th edition, Chapter 71, pp 1521-1551, Mc-Graw-Hill,
2001.
[0096] FIG. 4 shows how odd-carbon therapy serves to circumvent the
problems of glycogen metabolism that may be present. Administration
of odd-carbon fatty acids (for example as triglycerides or as acids
such as heptanoate) provides an alternate energy source, in the
form of intramitochondrial acetyl-CoA and Propionyl-CoA, that can
be utilized directly by the Krebs or "citric acid" cycle regardless
of the existence of any one of the defects of glycogen metabolism
(shown as red bars). Therefore, the demand for energy from glycogen
is eliminated.
[0097] FIG. 5 is a graph that shows the effect of a
triheptanoin-containing diet on the levels of select amino acids in
an adult onset glycogen storage disease--Type II patient (GSD II or
Pompes' Disease) following treatment with the Triheptanoin diet.
The characteristic deficits of Alanine and Glutamine were restored
within 13 hours with only the Triheptanoin diet, which documents
the rapid elimination of the need for degradation of body muscle
protein by provision of energy from Triheptanoin. The rapid
reduction of the serum levels of these amino acids is obvious when
the source of Triheptanoin was interrupted (NPO) awaiting the
placement of a gastrostomy tube. Following that event and the
resumed delivery of Triheptanoin via the gastrostomy tube, these
serum amino acids were rapidly restored to normal.
[0098] FIG. 6 shows the steps and location of, e.g., C7 metabolism.
The abbreviations on the right hand side of the arrows shows the
location of various FODs that are mitigated and/or avoided using
the odd chain fatty acids of the present invention as a single
source of fats or as a supplement. FIG. 7 summarized the "CAC
acceleration" of the present invention. By providing additional
C2-CoA and C3-CoA into the CAC, the odd chain fatty acids of the
present invention circumvent and avoid the enzymes that are
deficient or lacking in FOD patients. Importantly, the odd chain
fatty acids of the present invention may be provided as an adjunct
to current enzyme replacement therapies, e.g., intravenous
PEGylated-.alpha.-glycoside or other like enzymes. By providing
additional support for patients, a larger pool of patient may be
addressed, with reduced cost, increased compliance, reduced
morbidity and mortality and increased patient satisfaction.
[0099] The odd chain fatty acids of the present invention not only
prevented lipolysis they were also found to eliminate chronic
infections in FOD patients. To spare intra-mitochondrial Coenzyme
A, simple carnitine supplementation is required. Importantly, it
was found that the odd chain fatty acids optimized the supply of
propionate and acetate to the CAC and augmented ATP Production.
Furthermore, unlike the use of supplemental propionate, the present
invention did not sterilize the gut. The present invention provides
a readily available energy source via an alternate pathway and may
even be used to evaluate and protect siblings from disease.
[0100] The basic principle of the alternative metabolic pathway of
the present invention is to reduce the need for carbohydrate in the
form of stored glycogen or polysaccharide as substrate for the
Citric Acid (Kreb's) Cycle (CAC). Meeting the energy needs of the
body with Triheptanoin or other odd carbon fatty acids (C15, C5 or
BHP) can permit the gradual reduction of excessive stored glycogen
or polysaccharide resulting from any of the various types of
glycogen storage disease. This is accomplished by the intact
alternate metabolic pathway for glycogen degradation that exists in
either the cytosol and in the lysosome. Patients with GSD II must
remain on a diet containing odd carbon fatty acids, daily, for
life, in order to prevent further occurrence of the clinical
complications of the disease.
[0101] The amount of odd carbon fatty acid or ketone to provide is
25-35% of total Kcal/day for humans and .about.10-20% of total
Kcal/day for horses and other animals. The amount given can be
calculated based on the weight of the patient in kilograms (Kg).
The effective dose for adult humans and horses when triheptanoin is
utilized, for example, is 1-2 gm/Kg/day. The dose for children when
triheptanoin is utilized is approximately 1 to 4, preferably 3-4
gm/Kg/day.
[0102] The odd carbon fatty acid or ketone may be administered
enterally or parenterally. Enteral administration includes oral
administration and administration via a nasogastric tube or via a
gastrostomy tube. The administered substance may be in a food or
beverage or a nutritional composition. It may be desirable to
control or monitor the total caloric intake of the subject and this
may be accomplished through preparation of an appropriate liquid
diet. A parenterally administered preparation may be made by
methods known in the art for adding fatty acids as triglyceride
emulsions to parenteral nutritional preparations.
[0103] For horses as well as such animals as cattle and sheep, one
may use from about 1-2 gm/kg/day of triheptanoin and sufficient
amounts of any other odd carbon fatty acid or ketone to provide
about 10 to 20% of the total Kcal/day. The dose must be maintained
at the optimal level related to Kg body weight for the best
therapeutic result. This is especially important for a young
growing animal or human child. Symptoms return when the dose per
kg/day decreases as a result of growth and weight gain. It can be
given to animals in a variety of ways, as with humans. It can be
mixed with liquids, given straight via tube, or absorbed into "feed
pellets" like "alfalfa" or other animal feeds. Parenteral therapy
for animals is not a frequent treatment approach at this time, but
it could be used very effectively with horses and other large
animals when illness or stress results in serious metabolic
crisis.
[0104] Organic Acids may be measured as follows. Organic acids in
urine specimens are determined with isolation by Liquid Partition
Chromatography (LPC) and quantitative Gas Chromatography-Mass
Spectrometry (GCMS) using an internal standard. The specimen is
derivatized to form pentafluorobenzyl oximes (PFBO) of oxo-acids
and the organic acids isolated by Liquid Partition chromatography
on silicic acid hydrated with dilute sulfuric acid and eluted with
mixtures of tert-Amyl alcohol and chloroform. After drying, the
organic acids are derivatized to form volatile trimethylsilyl (TMS)
derivatives for separation by capillary Gas Chromatography (GC)
with temperature programming. Detection is by Mass Spectrometry
(MS) with identification of the organic acids by their mass
spectra. Organic acids are quantified by peak areas of
reconstructed ion chromatograms with internal standards and
calibration curves.
[0105] Acylcarnitine Profiles. Acylcarnitines and carnitine levels
in dried blood spots (dbs), plasma and amniotic fluid by automated
electrospray tandem mass spectrometry Acylcarnitines are extracted
from blood spots, plasmas and amniotic fluid with methanol
containing stable isotopically labeled internal standards. The
extract is derivatized to form butyl esters. An automated sample
introduction system delivers the samples to the electrospray tandem
mass spectrometer (ESI-MS/MS) operating in the multiple channel
analysis mode in which acylcarnitines are identified, profiled, and
quantified with stable isotopically labeled internal standards.
This test also provides quantification of free, sum of
acylcarnitines, and total carnitine levels.
EXAMPLE 1
Treatment of Human Subject Diagnosed with Acid Maltase
Deficiency
[0106] Clinical Description. A 42 year old caucasian female had
onset of progressive muscle weakness and difficulty breathing
dating back nearly 20 years. She was seen at 40 years for muscle
weakness and a muscle biopsy revealed increased glycogen deposition
in lysosomes and cytosol (2.0% --normal 1.03 +/-0.18%), autophagic
vacuolization, and increased acid phosphatase staining. An assay
for acid-.alpha.-glucosidase confirmed the deficiency as
adult-onset GSD II (0.46--normal 8.13 +/-2.1 nm
MU-hydrolyzed/min/gm). No therapy was initiated.
[0107] During the subsequent 2 years, the patient experienced
progressive muscle weakness, affecting strength and endurance,
swallowing and producing urgency for urination and defecation. The
patient's breathing became a major issue and weight had decreased
from .about.127 to 101 lbs (46 Kg) over this interval. Pulmonary
evaluation a day prior to admission revealed MIP=-70 cm HOH;
MEP=+60 cm HOH; FVC=40% of predicted (1.4 liters); blood gases: pH
7.38, pCO.sub.2 68, pO.sub.2 56 (room air). Home O.sub.2 was
started and upon respiratory arrest was intubated and
unsuccessfully extubated leading to emergency admission lasting one
month. Following informed consent, the patient agreed to enter an
approved Institutional Review Board (IRB) Protocol for dietary
therapy with triheptanoin oil.
[0108] Clinical and Biochemical Course on Therapy. The patient was
admitted with left lower lobe atelectasis and infiltrate, placed on
antibiotics, carnitine supplement (20 mg/Kg/day) and diet
containing triheptanoin was given by nasogastric tube. Esophageal
dysmotility was documented. A percutaneous endoscopic gastrostomy
(PEG) was placed on the sixth hospital day.
[0109] The diet was based on vivonex and .about.1500 Kcal was given
by nasogastric tube, on admission.
[0110] The composition (% Kcal) was: Protein 29%, carbohydrate
(CHO) 30%, total lipid 41% (of which triheptanoin (C7) accounted
for 26%). The C7 dose of 1 gm/Kg/day (8.3 Kcal/gm) was gradually
raised from 26% to 35% and finally 40% of total Kcal corresponding
to 1.0, 1.5, and 2.0 gm C7 per Kg per day. Carbohydrate was
correspondingly reduced from 30 to 27 to 11% of total Kcal/day.
Total caloric intake was raised from .about.1500 to 1700 to 1900
per day with these dietary changes during her 33 day admission.
[0111] Within 13 hours of beginning the C7 diet, pCO.sub.2 fell
from 68 to 43 (normal 35-45), and a long history of urgency for
urination and defecation had ceased and remained normal for the
rest of the admission. The patient began ambulating by the 10th day
that improved steadily, showering with assistance by the 28th day.
Pneumonia on day 17 responded to therapy and was extubated on day
23. She was discharged on bilevel positive airway pressure (BIPAP),
the C7 diet, and carnitine supplement.
[0112] Following discharge, the pateint returned to work half-time
at 5 weeks and was working full-time by 10 weeks. Swallowing was
sufficiently improved that gastrostomy feeds were stopped at 7
months and the PEG was removed at 10 months. By 15 months
post-discharge, patient weight had increased from 46 Kg on
admission to 60 Kg and has remained stable. Patient maintains
normal activities without any significant weakness but continues to
use BIPAP at night. Patient pCO.sub.2 levels remain in the normal
range, however, her full vital capacity remains at 40-45% of
predicted.
[0113] Metabolic Observations. Routine blood chemistries on
admission were largely normal except for pCO.sub.2 of 68.1 (normal
35-45 units). ALT and AST and LDL were mildly elevated while serum
CPK was normal. Hemoglobin was 11 gm/dl and creatinine was reduced
slightly at 0.4 (normal 0.5-1.0). Urinalysis revealed ketosis.
Potassium was reduced, slightly (3.5, normal 3.6-5.0 units). Blood
acylcarnitine analysis revealed normal total carnitine with
increased acetycarnitine (ketosis), and a surprising decrease in
propionylcarnitine (1.25, normal <2.71 uM). There were no other
acylcarnitine abnormalities.
[0114] Quantitative urinary organic acids revealed only ketosis
with a urine pH=8.5. The excretion of 3-OH-butyrate (BOB) and
acetoacetate (AcAc) was 1633 and 1211 mmol/mol creatinine,
respectively, (normal: <17 & 7 respectively). The ratio of
BOB:AcAc was decreased from 2.4 to 1.3 suggesting an alteration in
the intra-mitochondrial redox state. During hospitalization and
follow-up, this ratio was less than 1.17 (range 0.40-1.17) during 4
of 5 episodes of mild to moderate ketosis. However, more recently
with addition of an amino acid supplement to the C7 diet, the
levels and ratio became normal (3.0). Citric acid cycle
intermediate excretion fell into the normal ranges for adults.
[0115] The most significant abnormalities were observed with plasma
amino acid analysis. The levels of Alanine and Glutamine were low
compared to the adult normal ranges. While the levels of the
branched-chain amino acids (BCAA=Leucine, Valine, & Isoleucine)
were within the adult normal ranges. The reduced levels of Alanine
and Glutamine have been observed often in this disease and has led
to the high protein-low carbohydrate diet and/or alanine
supplementation. This patient did not receive any supplementary
alanine. In only thirteen hours after initiating the C7 diet by
nasogastric tube, the plasma alanine level had increased 4-fold.
Glutamine increased nearly 2-fold as did the branched chain amino
acids (BCAAs). The levels of these amino acids remained in the
normal range while receiving the C7 diet. However, the diet was
interrupted on day 4 & 5 while awaiting placement of the PEG
for gastrostomy feeding. The levels of each of these amino acids
promptly fell to admission levels (Table 2: 65 & 84 hour
samples). With resumption of the diet via gastrostomy, all levels
returned to (or above) the normal ranges in 24 hours. These amino
acid levels remained normal on this odd-carbon triglyceride diet
therapy. Further analysis of all of the measured amino acids
revealed the same sudden increase with the C7 diet and the same
sudden decrease when the diet was interrupted. As C7 was increased
from 26 to 40% of total caloric intake (from 1.0 to 2.0 gm/Kg/day).
All amino acids remained normal and propionylcarnitine levels had
increased without any evidence for propionyl overload from the
metabolism of heptanoate.
[0116] Furthermore, the present invention may be used to provide a
viable source of energy to patients with compromised ketogenesis,
gluconeogenesis and even the urea cycle, due to FODs. The odd chain
fatty acids help support the sub-optimal function of the Citric
Acid Cycle, gluconeogenesis, the urea cycle and ATP generation by
the respiratory chain. The anaplerotic substrates support the
Citric Acid Cycle, e.g., by providing pyruvate to malate and
oxaloacetate; glutamate to ketoglutarate and heptanoate to
propionyl-CoA to succinyl-CoA. These types of support may be
provided alone or in combination. Furthermore, the present
invention may be used with, e.g., amino acid supplementation to
reduce or prevent amino acid scavenging as an alternative source of
energy. As the patient improves, muscle and other tissues require
additional amino acid support, which may be supplemented along with
the present invention using amino acid supplements. One example of
an amino acid supplement for use with the present invention is
Amino-Vital, which contains branched chain amino acids (BCAA) via
branched chain ketone acids (BCKA) that may also supplement the
C2-CoA and C3-CoA. Additional co-factors and vitamins (e.g., Biotin
and B-12) may be added to the patient's diet to improve the
metabolism of Propionyl-CoA to Succinyl-CoA and/or entry into the
CAC as Acetyl-CoA.
EXAMPLE 2
Treatment of Human Subject Diagnosed with GSDII
[0117] Clinical History & Description. A 66 year old white male
whose history of muscle weakness dated back to his youth when he
noticed weakness that caused limitations in sports activities. In
his 40's, he became much more aware of weakness that primarily
affected his back and legs. Spirometry, at that time, revealed
reduced lung capacity. At age 54, a muscle biopsy was obtained that
showed glycogen deposition in lysosomes and cytoplasm but was not
increased (0.76%). Acid maltase was 0.94 (normal: 8.13 +/-2.1).
Debrancher enzyme was also assayed in this biopsy and was 1.40
(range 1.88-4.24). Shortly after that, he experienced shortness of
breath at high altitude and has used bilevel positive airway
pressure (BIPAP) since that time also for sleep apnea. Patient
muscle weakness was progressive and he has been using a motorized
scooter for the past 10 years. Patient weakness was generalized, he
had great difficulty standing, and walked with an irregular gait,
using a cane. Patient had no history of impaired swallowing but did
complain of urgency for urination and defecation.
[0118] Clinical Course. Following informed consent, the patient
agreed to enter an approved IRB Protocol for dietary therapy with
triheptanoin oil. Initial pulmonary studies revealed severe
restriction of his vital capacity (2.41 L, 43% of predicted).
Maximal inspiratory and expiratory pressures were 60 cm of water.
Initial arterial blood gases revealed pH=7.39, pCO.sub.2=51. And
pO.sub.2=62.
[0119] The oral diet consisted of .about.2900 Kcal on admission.
The composition (% Kcal) was: Protein 16%, carbohydrate (CHO) 40%,
total lipid 44% (of which triheptanoin (C7) represented 22%). The
C7 dose of 1 gm/Kg/day (8.3 Kcal/gm) was gradually raised from 26%
to 30% of total Kcal corresponding to 1.0, and 1.5, gm C7 per Kg
per day. Carbohydrate was correspondingly reduced from 43 to 22% of
total Kcal/day. Total caloric intake was maintained at .about.2800
per day during his 30 day admission.
[0120] Initially, the patient required assistance and supervision
with grooming, upper and lower extremity dressing, bathing, and
toileting. The patient also required maximal assistance with
ambulation and transfers from bed to wheelchair. With intensive
physical therapy and diet, at discharge, he had became independent
with grooming, upper and lower extremity dressing, toileting, and
locomotion with a manual wheelchair. He continued to require
supervision, but without assistance, with transfers from bed to
wheelchair and to toilet and shower. He had clearly made
improvements in strength and endurance with exercise. Pulmonary
function testing showed an increase of 700 ml in total vital
capacity when compared to his studies on admission.
[0121] The patient was discharged on the triheptanoin diet with
supplements of citrate (bicitra, 15 cc four times a day) and amino
acid supplement ("Amino-Vital", Ajinomoto, Inc) 4.8 gms four times
a day. At four months after discharge, he continues this regimen
and physical therapy three times per week.
[0122] Metabolic Observations. Routine blood chemistries on
admission were largely normal except for pCO.sub.2 of 51 (normal
35-45 units). ALT and AST and lipid profile were normal while.
serum CPK was mildly increased (218, normal: 38-174 IU/L). His BUN
was increased (30, range 9-20 units) and creatinine was reduced
slightly at 0.6 (normal 0.7-1.2 units). All other chemistries were
normal. Blood acylcarnitine analysis revealed normal total
carnitine and a reduced level of propionylcarnitine (1.08, normal
<2.71 uM). There were no other acylcarnitine abnormalities.
[0123] Patient diet was initially adjusted to 2800-3000 Kcal/day
composed of protein 16%, CHO 40%, total lipid 44% of which C7
represented 22% of his daily Kcal intake. The C7 composition was
increased to 30-31% during the admission. His initial response to
C7 reveled a marked decrease in urinary pH (from 7.5 to 6.0) and
acylcarnitine monitoring revealed a marked increase in
propionylcarnitine (1.08 to 5.86 uM) and pimelate, heptanoate and
methylmalonate (MMA) excretion increased significantly (321, 307,
and 38 mol/mol creatinine, respectively. These changes indicated
impaired oxidation of C7 and reduced utilization of propionyl-CoA
for the citric acid cycle
[0124] (CAC). Following supplementation with citrate (bicitra) the
urine pH returned to 7.5, and pimelate, heptanoate, and MMA
decreased immediately to 58, 50, and 24, respectively. The
propionylcarnitine blood level was also reduced to 3.74 uM. These
changes reflected more normal metabolism of dietary C7. Citrate
supplementation was continued throughout the treatment.
[0125] Although there were 5 periods when the quantitative urinary
organic acids revealed mild ketosis, the ratio of BOB:AcAc was
always normal while on citrate supplementation. Unlike patient 1,
there was no indication of an alteration in the intra-mitochondrial
redox state. Although citric acid cycle intermediate excretion was
in the normal adult ranges, there were some abnormalities:
isocitrate was initially increased (134-n1=<82) while succinate
and u-ketoglutarate levels were reduced (6 [n1 <42], and 4
[n1<75], respectively. The sum of the excretion of the CAC
intermediates was 391 (upper limit=1185). This profile persisted
during the study.
[0126] The most significant abnormalities were observed with plasma
amino acid analysis. The levels of Alanine and Glutamine were low
compared to the adult normal ranges. While the levels of the
branched-chain amino acids (BCAA=Leucine, Valine, & Isoleucine)
were within the adult normal ranges. The reduced levels of Alanine
and Glutamine have been observed often in this disease and has led
to the high protein-low carbohydrate diet and/or alanine
supplementation. This patient did not receive any supplementary
alanine. In only thirteen hours after initiating the C7 diet by
nasogastric tube, the plasma alanine level had increased 4-fold.
Glutamine increased nearly 2-fold as did the BCAA's. The levels of
these amino acids remained in the normal range while receiving the
C7 diet. However, the diet was interrupted on day 4 & 5 while
awaiting placement of the PEG for gastrostomy feeding. The levels
of each of these amino acids promptly fell to admission levels.
With resumption of the diet via gastrostomy, all levels returned to
(or above) the normal ranges in 24 hours. These amino acid levels
remained normal on this odd-carbon triglyceride diet therapy.
Further analysis of all of the measured amino acids revealed the
same sudden increase with the C7 diet and the same sudden decrease
when the diet was interrupted. As C7 was increased from 26 to 40%
of total caloric intake (from 1.0 to 2.0 gm/Kg/day). All amino
acids remained normal and propionylcarnitine levels had increased
without any evidence for propionyl overload from the metabolism of
heptanoate.
EXAMPLE 3
Treatment of Horses
[0127] Three horses affected with biopsy proven, polysaccharide
storage myopathy have been successfully treated with the
triheptanoin diet. The first was an 8 year old mare who was unable
to walk or exercise without severe muscle pain, rhabdomyolysis, and
"locking up." The owners were approaching "putting the animal down"
(killing it). Within 30 days of triheptanoin diet incorporated into
feed pellets, this animal has become asymptomatic and has returned
to full training and riding without any symptoms.
[0128] A second horse was encountered at 4 months with proven
polysaccharide storage myopathy (PSSM) that was not able to walk
adequately or nurse from its mother. The owners were prepared to
kill the animal but decided to try the triheptanoin diet first.
Within 24 hours of initiating the triheptanoin diet, the foal was
running and playing with other young horses without any evidence of
muscle pain, etc. All blood enzyme levels related to muscle disease
(including serum creatine phosphokinase) had normalized. As this
foal gained weight, in a normal rate for a young horse, symptoms
returned since the dose of C7 had not been adjusted for the weight
gain. Adjustment of the dose back to 2 grams/Kg body weight
eliminated all symptoms and there is no further evidence of
disease.
[0129] A third horse, also an 8 year old mare, developed PSSM and
"locked up." She was no longer available to ride and could no
longer jump. She started triheptanoin at 1.0 gm/Kg/day added to her
diet pellet feeds and is currently asymptomatic with normal blood
chemistry levels compared to pre-treatment after only one week of
diet treatment.
EXAMPLE 4
Treatment of FOD in Horses and Diets for Treatment of Fatigue
[0130] Horses: Eight healthy, fit 4 year old thoroughbred geldings
may be used in a study. These horses are all sound and trained to
the treadmill. The ability of horses to absorb and digest C7 oil
was first determined. Three horses were fasted for 12 hours prior
to administration of 0.5 gm/kg of triheptanoin via nasogastric
tube. Samples for blood acylcarnitine profile were drawn at 30, 60,
90, 120, 150, and 180 minutes after triheptanoin administration. C7
acyl carnitine peaked at 90 minutes after administration and peak
serum concentration of C5 and C3 acylcarnitines occurred 120
minutes after administration (FIG. 9).
[0131] In humans C7 (triheptanoin) is hydrolyzed to heptanoate
which is then oxidized in the liver to form .beta.-hydroxypentonate
and .beta.-ketopentonate, both 5 carbon ketone bodies (C5). C5 is
metabolized to C3 (propionyl CoA) and C2 or acetyl CoA. These fatty
acids are transported in the bloodstream as acylcarnitines, which
are easily taken up in muscle tissues and cross mitochondrial
membranes to enter the CAC.
[0132] The five carbon ketones .beta.-hydroxypentonate and
.beta.-ketopentonate produced from the breakdown of triheptanoin in
the liver are represented in our preliminary results as plasma C5
acylcarnitine. Horses have a poorly developed ability to produce
ketone bodies, however the presence of 5 carbon acylcarnitines in
our horses, suggests that they were able to metabolize triheptanoin
into the 5 carbon ketone. C3 acylcarnitines were also increased in
our preliminary studies suggesting the break down of the C5 to
propionyl CoA (C3).
[0133] Palatability of C7 has also been studied, in which the C7
was provided as triheptanoin or mixed with corn oil into the diet
of horses. It was found that when horses were fed the oil mixed
into hay cubes soaked in water prior to feeding, they consistently
ate all of the feed.
[0134] Diets: A variety of diets may be formulated. For example,
the diets may be isocaloric and formulated to meet the minimum
daily requirements for all nutrients. In one example of horse feed
is made that includes 8.5 kg of grass hay/grass hay cubes, 0.5 kg
of ration balancer, free choice salt and 750 mls of oil per day
with a total digestible energy of 23 MCal/500 kg horse per day. The
oil will be either triheptanoin (provided by Dr. Roe) or corn oil
(purchased) and will be divided into 3 feedings.
[0135] Exercise study: Horses will generally be rested for one day
prior to and one day after the exercise study. On day 14 of each
diet an IV catheter will be placed and horses will perform a 90 min
submaximal exercise study on a flat treadmill 120 min after
consuming the oil. A loosely fitting mask with a high flow rate may
be placed over the horse's nose and samples of expired gas
collected every 15 minutes during exercise using an open circuit
calorimeter. Oxygen consumption (VO.sub.2) and carbon dioxide
production (VCO.sub.2) will be measured and respiratory exchange
ratio (RER) will be calculated.
[0136] The exercise study protocol may include a 5 min warm up at a
walk, followed by: [0137] Block 1: 15 min 35% VO.sub.2 max
(previously measured in these horses), 5 min walk [0138] Block 2:
15 min 35% VO.sub.2 max, 5 min walk [0139] Block 3: 15 min 35%
VO.sub.2 max, 5 min walk [0140] Block 4: 15 min 35% VO.sub.2 max, 5
min walk [0141] Block 5: 10 min 75% VO.sub.2 max [0142] Block 6: 5
min walk
[0143] Blood samples: Blood samples may be drawn prior to the
initiation of the trial; prior to the exercise study; and at the
end of the highest speed during each exercise block of the exercise
study. Blood samples for glucose, lactate, ketones, FFA, amino
acids and acylcarnitine concentrations will be obtained. Glucose
and lactate will be assayed on an automated lactate analyzer and
ketones and FFA will be assayed spectrophotometrically using the
appropriate kits. Plasma amino acid and aclycarnitine levels may
also be measured by mass spectrometry from four randomly selected
horses; all other values will be analyzed in all study
subjects.
[0144] Muscle Biopsy: Gluteal muscle biopsy specimens may be
obtained by use of a percutaneous needle biopsy technique prior to
initiation of the diet trial; prior to and immediately upon
completion of the exercise study; and 24 hours after exercise
study. Samples are obtained within a 2 inch square at a
standardized site along the gluteal medius muscle and alternating
sides will be used for each sample. Biopsy specimens will be
immediately frozen in liquid nitrogen and stored at -80.degree.
C.
[0145] Muscle Analysis: Frozen muscle specimens will be dissected
free of blood and connective tissue. Glycogen will be assayed
flourometric as glucose residues remaining after 1-2 mg portions of
muscle tissue are boiled for 2 h in 1 M HCl. Lactate, pyruvate,
G-6-P, and CAC intermediates will be assayed in a separate portion
of the muscle sample (8). One 4 mg portion of muscle will be
homogenized by crushing with a glass rod in 1.5M perchloric acid.
The supernatant obtained after centrifugation will be neutralized
with KHCO.sub.3, centrifuged again and the remaining supernatant
removed for analysis of metabolites using fluorometric techniques
and purine nucleotides using high-performance liquid chromatography
(HPLC). Concentrations of CP, ATP, ADP, AMP and IMP in muscle
specimens as well as in external standards will be analyzed by use
of a reverse phase column (10). Separation of nucleotides will be
achieved with a flow rate of 1.0 mL/min, UV-detection at 254 nm,
and an oven temperature of 40.degree. C.
[0146] Urine Samples: A urine collection device is placed the
evening prior to exercise on the 4 horses randomly selected for
plasma acylcarnitnies and amino acid analyses and will again be
placed on the horses after exercise. An aliquot of voided urine
will be collected for measurement of urine organic acids by mass
spectrometry.
EXAMPLE 4
Beverages and Food Supplements
[0147] In one embodiment, the odd chain fatty acids of the present
invention may be provided as a nutritional supplement and/or as
part of an overall patient diet. As a non-limiting example of a
foodstuff that may include the present invention in a
pharmaceutical or nutritional amount, the odd chain fatty acids may
be formulated into a snack bar, such as that described in U.S. Pat.
No. 4,777,045, issued to Vanderveer, relevant portions incorporated
herein by reference. A high bran snack is taught that includes a
typical formulation of graham (whole wheat) flour, 40 percent; rice
flour, 10 percent; whole wheat bran flour, 50 percent; calcium
carbonate, 1.25 percent; reduced iron, 0.013 percent; and
riboflavin (as a tracer), 0.02 percent and between 0.1 and 10% odd
chain fatty acids. Water is added per 80 ounces of dry materials to
provide a semi-solid flowable product that may be fed into a ribbon
blender and then into a cooker extruder, e.g., a twin-screw cooker
extruder. Each of the ingredients in the formulation has a
processing, nutritional or therapeutic purpose. The extruded pieces
may be further flavored, e.g., coated with coconut oil and powdered
flavorant.
[0148] A whole wheat and bran snack may be 50 weight percent of
wheat bran, with approximately 50 percent of whole wheat flour,
including 0.2 weight percent of riboflavin (20 mg per 100 gm of
finished product), have a desired size, density and shape for the
subsequent operation of adding oil, flavor and eventually
packaging. Samples of finished products, with 4 different flavors,
applied at a level of 3 weight percent to the extruded base
product, with the addition of 3 weight percent of odd chain fatty
acids. The extent and formulation of the odd chain fatty acids
(e.g., with an emulsifier) may improved adhesion of the flavor
particles, flavor acceptability and mouth feel organoleptically.
The bar may further include vitamins, minerals and even protein and
carbohydrates to provide a so-called "power-bar."
[0149] Another bar may be a granola bar with supplemental dietary
fiber as taught by Linscott in U.S. Pat. No. 4,871,557, relevant
portions incorporated herein by reference. A granola bar is taught
that includes supplemental dietary fiber added to the granola bar
in the form of compressed flakes, as well as the method of making
such a granola bar, which may further include the odd chain fatty
acids of the present invention. Briefly, a mixture of granola
ingredients such as grains, fruits, nuts and compressed flakes are
mixed with the granola ingredients and the odd chain fatty acids.
The compressed flakes of supplemental dietary fiber are combined
with water and a binder material, such as rice flour, and then
extruded. The extrudate is dried and then ground to the desired
particle size.
[0150] Although sources of supplemental dietary fiber can
contribute both soluble and insoluble fiber, sources generally
known to contribute insoluble fiber include but are not limited to
soy fiber, apple fiber, corn bran, wheat bran, oat bran, barley
bran, rye bran, triticale bran, cellulose, pea fiber, sugar beet
fiber, and peanut fiber. Sources generally known to contribute
soluble fiber include but are not limited to gum arabic, gum
ghatti, guar gum, pectins, psyllium, carrageenans, xanthan,
tragacanth, karaya, locust bean gum, agar, and alginates. These
non-digestible fibers provide additional control over the exact
amount of kilocalories are provided to a patient as the primary
source of energy from fat is provided by the odd chain fatty
acids.
[0151] The extrusion step may be performed by conventional
techniques in conventional extrusion apparatus, e.g., a damp
mixture is heated to a temperature between about 300 and about
330.degree. F., e.g., 315.degree. F. during the extrusion process.
The damp mixture is then extruded at a pressure of between about
100 and about 900 p.s.i. A die through which the mix is extruded
may includes a square, rectangular, triangular, oval or round hole
with a diameter of, e.g., 0.5 inches. Generally, extrudates are
particle sized before being dried in an oven at 270.degree. F. for
about 20 minutes. Often, the dried particles will have a moisture
content of about 7 percent.
[0152] The odd chain fatty acids may also be provided as beverage.
The odd chain fatty acids may be added to a beverage such as that
taught in U.S. Pat. No. 4,981,687, relevant portions incorporated
herein by reference. The beverage taught is used to improve
physiological responses to exercise, however, it uses standard
sources of carbohydrates. The addition of the odd chain fatty acids
taught herein provides a beverage that not only reduces or prevents
adverse physiological effects of physical exercise or environmental
exposure, but provides a new and/or additional sources of readily
available energy. The fluid will generally include water,
electrolytes, and odd chain fatty acids and is non-toxic to man or
animals. Sugar may also be provided or sugar substitutes may be
used without a concomitant loss of new energy production. As such,
the odd chain fatty acids substitute the energy source and can be
absorbed rapidly through the gastrointestinal tract, prevents
decreases in blood volume.
[0153] The odd chain fatty acids will provide a beneficial
physiological effect on cells during exercise by providing a
substitute and/or additional readily available energy source that
may increase blood volume and cardiac output, improved skin blood
flow, prevention or delay of onset of hyperthermia, increased rate
of movement of electrolytes across the gastrointestinal wall,
reduction in the breakdown of proteins and associated metabolism of
essential amino acids, and decreased time needed for repair of body
tissue following strenuous exercise.
[0154] When the fluid of the subject invention is administered, the
body's physiological response to exercise or environmental exposure
is greatly enhanced compared to the response when the body receives
no fluids, receives only water, or receives fluid such as
GATORADE.RTM., which contains electrolytes and a sugar source in
addition to water. The composition described here can be used to
ameliorate the adverse effects of physical exertion or
environmental exposure. The effect of the odd chain fatty acids on
physical exertion may be measured by determining plasma volume,
respiratory quotient, rectal temperature, pulse rate and/or cardiac
output; combined with either enhanced endurance or performance,
lower perceived difficulty of a physical task, or an enhanced
ability to withstand heat exposure or chronic exposure to cold. The
beverage may include: water, odd chain fatty acids (1-4%),
potassium (2 meq/l), sodium (26 meq/l) and phosphate (4 meq/l).
[0155] Addition of a small amount of odd chain fatty acids, given
at frequent intervals, may improve performance and endurance
because it enhances entrance of both acetyl CoA and propionyl CoA
into the Krebs cycle, thereby bypassing the need for additional
oxaloacetate (OAA). The Krebs cycle is a well-known, but very
complicated, biochemical pathway which provides a working muscle
with its energy source. A detailed description of the Krebs cycle
can be found in most biochemistry textbooks.
[0156] The beverage of the present invention may also be used to
improve the physiologic response of any animal undergoing exercise
or being subjected to high temperature conditions. For example,
humans, horses, dogs, cats, mules, oxen, camels, emu, bison,
nilguy, elephants, sheep, cows, chickens, turkey, goats, llamas,
alpaca and pigs are a few of the animals that may benefit from the
administration of the novel fluid composition described here. The
fluid of the subject invention can also be used to alleviate or
prevent dehydration which is known to result from chronic exposure
to cold temperatures.
[0157] The odd chain fatty acids of the present invention may also
be provided with dietetic beverages. An example of a dietetic
beverage that may include the odd chain fatty acids includes, e.g.,
U.S. Pat. No. 4,042,684 that teaches a beverage for supplementing
the dietetic requirements of sugar and essential salts in a
mammalian body depleted through vigorous physical activity. The odd
chain fatty acids may be added and substitute for simple
carbohydrates in an aqueous that includes sodium chloride,
potassium chloride, and free citric acid in physiologic ranges.
[0158] The combination of odd chain fatty acids and essential salts
in a mammalian body will be particularly useful for athletes that
have depleted short-term, immediately available energy through
vigorous physical activity. For example, an athlete engaged in
strenuous activity requires a ready source of energy for endurance,
and replacement of both body fluids and essential salts lost
through perspiration. Likewise, individuals working in a hot, humid
atmosphere have similar requirements to maintain efficiency and
productivity. Long term activity initiates bypass metabolism that
the odd chain fatty acids of the present invention help to
supplement to increase protein sparing.
[0159] Typical diets and sports beverages provide intake of sugars
and sugar-precursor materials such as carbohydrates. When
metabolized in the digestive tract, fuel values are obtained
through enzyme and acid attack on these complex sugars. However,
the process requires time to achieve the desired boost in
metabolism, which is short lived and leads to protein and/or amino
acid scavenging to supplement the citric acid cycle. Increased
metabolic demands exceed the steady-state ability of this natural
metabolic process.
[0160] Rather than rely on well-known sugar metabolism, e.g.,
fructose and glucose to achieve an energy store capable of
providing needed fuel values, odd chain fatty acids provide the
fully utilizable energy at the cellular level for metabolism in
either the liver or muscles, brain, kidney and heart. One problem
with commonly available sports beverages is that, e.g., glucose is
easily and rapidly transported out of the digestive system into the
blood whereas fructose is more passively and slowly transported.
Once into circulation, the fructose is somewhat more efficient
insofar as initial transport requires less energy and subsequent
utilization for energy proceeds more readily. Thus, both immediate
and longer lasting benefits may be attainable. The odd chain fatty
acids disclosed herein may be provided with or partial or complete
substitution for those complex saccharides that have differential
breakdown and uptake. The odd chain fatty acids are provided along
with salt constituents, either as salts with the odd chain fatty
acids themselves, or in the form of additional salts, e.g., sodium
chloride and potassium chloride. The quantity and relative
proportion of the salts may be selected to achieve an isotonic
liquid without a strong acid taste. Thus, the salt components
provide replacement for those essential ions lost in perspiration
while, at the same time, yielding a highly palatable beverage.
[0161] The beverage of the present invention may be compounded with
water and suitably bottled and stored. Alternatively, all the
components, save for water, may be prepared in advance as a
concentrate for ease of handling and transportation. Also, the
beverage may be prepared with carbonated water should such be
desirable.
[0162] During athletic competition, or other strenuous physical
activity, the individual may replace lost body fluids and essential
salts while sustaining a high level of energy through consumption
of the beverage of the present invention. Moreover, contrary to
many of the prior art formulations for such beverages, e.g.,
extended citrus juices, intake of the beverage of the present
invention is not accompanied by a "full" feeling that would present
a serious hindrance in athletic competition. Thus, the present
dietetic beverage successfully replaces and/or maintains individual
body requirements in accordance with the physiological needs
thereof and further provides additional storage of energy.
[0163] Food Compositions and Additives. The odd chain fatty acids
of the present invention may also be formulated into an edible
matrix and/or food composition. One such chewable matrix is taught
in U.S. Pat. No. 6,723,358 that teachers the encapsulation of
edible products, relevant portions incorporated herein by
reference. An encapsulated product is obtained by mixing a
plasticizer, a ground, free-flowing particulate mixture with the
odd chain fatty acids, at least one starch, mixed and heated. A
chewable texture is obtained when the starch is maintained
substantially ungelatinized. A flavorful product may also be
obtained and made part of an overall patient diet without
destroying a heat sensitive encapsulant because the oil and starch
are heated to develop flavor at high temperatures prior to mixing
with a heat sensitive encapsulant. The encapsulated component may
be at least one biologically active component, pharmaceutical
component, nutraceutical component, or microorganism. The mixture
may includes ground cookies that are mixed with the plasticizer,
odd chain fatty acids and water to obtain a formable dough or
crumbly mass that may be further formed into pieces or pellets and
dried to a shelf-stable moisture content.
[0164] The present invention will find particular utility in animal
feeds. For example, the odd chain fatty acids may be used in
conjunction with the feed stocks, e.g., that taught in U.S. Pat.
No. 6,777,396, relevant portions incorporated herein by reference.
The feed for livestock may include a feed for livestock with
additives such as nucleic acids, glutamine and glutamic acid. The
feed is provided as part of a method for increasing body weight
gain efficiency and feed efficiency in livestock.
[0165] The present invention also includes an animal feed
composition that includes the odd chain fatty acids. It is
contemplated that any odd chain fatty acids may be present in the
animal feed in any amount effective to provide nutritive fat to the
animal: It is contemplated that the odd chain fatty acids content
may vary depending upon the animal or upon the intended nutritive
qualities of the feed. Generally, the fat is present in the animal
feed in an amount of at least 5%, 10%, 17%, 20%, 25%, 30% or even
40% by weight of the animal feed. It is contemplated that two or
more odd chain fatty acids sources may be included in the feed.
[0166] The animal feed further includes a solid nutritive source,
such as a whole grain, whole wheat, whole rice, whole corn, or
whole barley. Alternatively, the solid nutritive source may also
include nutritive wheat, nutritive rice, nutritive corn or
nutritive barley fraction. Other nutritive sources include those
derived from soy, oats, sorghum, and the like. The nutritive source
may include other nutritive sources, including sources of
carbohydrates (such as molasses solids) that are also provided in
liquid form. The solid nutritive source may be present in the
animal feed in any suitable amount.
[0167] Another nutritive source is a protein source, which may be
present in any amount effective to provide protein to the animal.
Protein may be present in an amount ranging from about 5% to about
40% by weight of the animal feed. For example, young swine are
particularly needy of protein, and protein contents in the upper
portion of this range (e.g., a protein content of about 36%) are
often used in feeds intended for such swine. For feeds of other
animals, the protein is present in an amount ranging from about 10%
to about 30% by weight of the animal feed, but more often in an
amount ranging from about 15% to about 20% by weight of the animal
feed.
[0168] Animal feeds may also include a source of fiber source.
Sources of fiber may include:
[0169] soybean hulls, rice hulls, corn hulls, cottonseed, wheat
hulls, and the like are considered largely non-nutritive (at least
in the case of non-ruminant animals). When the animal feed is
intended for use by ruminants, the feed very often includes such
fiber source in an amount effective to provide fiber to the animal.
Different feed formulas for different animals vary greatly in the
amount of fiber desired. The fiber source may be prepared in an
amount ranging from about 1% to about 25% by weight of the animal
feed, the percentage being expressed by the bulk weight of the
hulls or other source.
[0170] Pelleted High-Fat Animal Feed. The odd chain fatty acids of
the present invention may be incorporated into any of a number of
animal feeds. Basic types of animal feeds are taught in, e.g., U.S.
Pat. No. 6,746,698, relevant portions incorporated herein by
reference, which teaches an animal feed, method for preparing
animal feed, and method for feeding an animal. Unlike the present
invention, however, the fats taught are standard even chain fatty
acids and fats. A master batch of horse feed ration containing full
fat corn germ from a corn wet-milling process was formulated in a
Hobart mixer. The feed was formulated from the following
components:
TABLE-US-00014 Ground Full-Fat Corn Germ 24.8% Wheat Midds 21.1%
Ground Whole Corn 17.9% Soybean Meal 9.5% Odd Chain Fatty Acid
25.0% Calcium Carbonate 1.1% Dicalcium Phosphate 1.1%
[0171] To an aliquot of the master batch an adhesive may be added,
in liquid and/or dry form in an amount sufficient to constitute 5%
of the total dry mass of the final pelleted feed. The mixture was
then converted into a pelletized horse feed using a pellet
mill.
[0172] An animal feed was formulated as follows:
TABLE-US-00015 Ingredient Weight % Corn Germ 23% Wheat Midds 21.5%
Whole Ground Corn 15% Soybean Meal 9% Calcium Carbonate 1%
Dicalcium Phosphate 1% Odd Chain Fatty Acid 25.0%
[0173] The ingredients may be combined to form a mixture and the
mixture pelletized. In this formulation, the wheat midds were
included to provide a source of fiber, the soybean meal was
included to provide a source of protein, and the calcium carbonate
and dicalcium phosphate were included as mineral calcium sources.
The feed may be evaluated for palatability.
[0174] A master batch horse ration formula having the following
composition was prepared:
TABLE-US-00016 Full-Fat Corn Germ 26% Wheat Midds 21% Ground Corn
10% Soybean Meal 6.5% Distillers Dried Grain 2% Calcium Carbonate
1% Dried Whole Wheat 1% Dehydrated Alfalfa 1% Dicalcium Phosphate
1% Odd Chain Fatty Acid 20.0%
EXAMPLE 5
Use of Odd Chain Fatty Acids to treat Mitochondrial Fat Oxidation
Defects
[0175] Other examples of FODs that derive from errors in metabolism
include: Carnitine Palmitoyl
[0176] Transferase I (CPT I) deficiency, Carnitine-Acylcarnitine
Translocase (CACT) deficiency, Carnitine Palmitoyl Transferase I
(CPT II) deficiency, very long chain acyl-CoA dehydrogenase (VLCAD)
deficiency, Trifunctional Protein (TFP) deficiency, long chain acyl
3-hydroxy CoA dehydrogenase (LCHAD) and short chain acyl CoA
dehydrogenase (SCAD) deficiency. Patients having each of these
diseases were located and placed on an odd chain fatty acid diet,
with informed consent and following under IRB. These patients
exhibit one or more of the following symptoms: recurrent
hypoglycemia (Reye-like); cardiomyopathy: hypertrophic or dilated;
hypotonia and/or delayed development; rhabdomyolysis; muscle
weakness and fatigue; peripheral neuropathy; retinopathy; seizures
and often sudden death due to, e.g., arrhythmias.
[0177] The odd chain fatty acid diet and therapy of the present
invention was compared to conventional treatments, which include:
(1) a low fat, high carbohydrate diet; avoid fasting; frequent or
continuous feeding and night time corn starch; or (2) a medium
chain triglyceride (MCT oil) for long chain disorders; or (3)
insulin and glucose to inhibit lipolysis. However, current
treatments fail to treat the hypoglycemia and hepatomegaly (which
often persist), the frequency and severity of
[0178] Rhabdomyolysis; muscle weakness and fatigue are not improved
and heart function often remains abnormal. Biochemically, the
failure of existing therapies leave patients with impaired
Acetyl-CoA production and ketogenesis, there remains an increase
Acyl-CoA:CoA (mitochondrion) ratio during illness, a decreased
NADH:NAD ratio during illness (mitochondrion), hypoglycemia and
hyperammonemia.
TABLE-US-00017 TABLE 3 Triheptanoin Patient Demographics. # Male/
FOD Patients Female Age @ Entry *Ethnic CPT I 2 1/1 4 yr 9 mo-6 yr
11 mo 2C CACT 1 0/1 birth 1H CPT II 7 2/5 5 yr 6 mo-51 yrs 4C, 3J
VLCAD 19 9/10 Neonate-35 yrs 16C, 1AA, 1ME, 1H LCHAD 9 4/5 6 mo-23
yrs 6C, 2H, 1J TFP 5 5/0 32 mo-8 yr 6 mo 5C "SCAD" 5 5/0 19 mo-6 yr
9 mo 3C, 2J TOTAL: 48 *C = caucasian, H = hispanic, J = jewish, AA
= afro-american, ME = middle eastern
[0179] FIG. 9 is a graph that shows the effects of triheptanoin on
patient creatine kinase activity for selected patient populations.
Tri-C7 rapidly suppresses CPK levels in long-chain defects because
it provides fuel to muscle via 5-carbon ketone bodies, thereby
giving propionate and acetate into the CAC, with an increase in
energy.
[0180] Table 4 summarized a comparison of patent mortality for
patients with FOD of conventional treatment (based on the
literature, see *Neonatal onset; Conventional Diet Rx: JIMD 22:
488,1999), and the odd chain fatty acids of the present invention.
There is a demonstrable increase in patient survival across a
number of FODs.
TABLE-US-00018 TABLE 4 Patient Mortality Comparison Conventional
Diet v. C7 Diet Conventional Diet (41 C7 Diet (48 patients - 6%
Disorder patients - 51% withdrawl) withdrawl) CACT* 5/5 - 100% 1/1
- 100% (rotavirus) CPT II* 4/5 - 80% No patients VLCAD 6/8 - 75%
1/19 - 5% TFP 4/4 - 100% 1/5 - 0% LCHAD 2/10 - 20% 0/9 - 0%
[0181] The following generalized clinical observations demonstrate
the effects of the odd chain fatty acids therapy on the patients.
First, sudden weight gain was at first welcome and prevented by
reduced carbohydrate intake. The use of emulsifiers and recipes
that incorporated the odd chain fatty acids into foodstuffs reduced
gastrointestinal (GI) intolerance. As regards chronic (occult)
infection a vast improvement was noted and rhabdomyolysis was
relieved. In a couple of cases a mild toxicity was noted and
resolved by the occasional increase in biotin or B12 requirement
and was overcome by supplementation in adults, only. Importantly,
no "Propionyl Overload" was observed in any patient. Compliance was
very good with the minimal withdrawal of 3 (6%) of 51 total
patients that withdrew due to complains of GI discomfort and/or
weight gain. Most importantly, the vast majority of patients
elected to continue beyond protocol: 22 (85%) of 26.
[0182] Regarding protein sparing, the following table summarized
the results obtained in a patient with Pompe's upon
TABLE-US-00019 TABLE 5 Plasma Amino Acids (.mu.M) During Initiation
of Anaplerotic Therapy: ''Protein Sparing'' in Adult-Onset Glycogen
Storage Disease II (Pompe's) Event: Admit C7 = 1.0 gm/Kg NPO* C7 =
1.5 gm/Kg Time: Baseline 13 hrs 41 hrs 65 hrs 84 hrs 108 hrs 132
hrs ''Oxaloacetate'': ASPARTATE (0-10) 8 44 112 15 12 50 42 ''Urea
Cycle'': ORNITHINE (36-118) 35 78 163 59 50 122 95 CITRULLINE
(11-51) 20 1 43 21 20 35 25 ARGININE (20-122) 50 96 93 71 84 102 97
''Neurotransmitters'': **PHENYLALANINE (35-83) 62 101 156 69 51 90
84 TYROSINE (30-100) 55 110 62 44 39 56 46 **TRYPTOPHAN (23-86) 25
121 43 20 <10 40 40 Total Amino Acids: (1540-4415) 1927 3673
4160 2176 2174 3568 3296 *NPO for gastrostomy - IV glucose only;
**Essential amino acids
[0183] In conclusion, the odd chain fatty acid diet and therapy of
the present invention was found to successfully treat the following
FOD disorders: CPT I, CACT, CPT II, VLCAD, TFP, LCHAD and SCAD. The
outcome of dietary triheptanoin supplementation included: reduced
mortality, cardiomyopathies were resolved and there was a marked
attenuation of Rhabdomyolysis. Also, normal glucose homeostasis was
observed with the elimination of hepatomegaly. Patients showed
improved muscle strength and endurance. In TFP peripheral
neuropathy was unchanged and in LCHAD the retinopathy
(Choroideremia) was also unchanged.
[0184] It will be understood that particular embodiments described
herein are shown by way of illustration and not as limitations of
the invention. The principal features of this invention can be
employed in various embodiments without departing from the scope of
the invention. Those skilled in the art will recognize, or be able
to ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures described herein. Such
equivalents are considered to be within the scope of this invention
and are covered by the claims.
[0185] All publications and patent applications mentioned in the
specification are indicative of the level of skill of those skilled
in the art to which this invention pertains. All publications and
patent applications are herein incorporated by reference to the
same extent as if each individual publication or patent application
was specifically and individually indicated to be incorporated by
reference.
[0186] In the claims, all transitional phrases such as
"comprising," "including, " "carrying," "having," "containing,"
"involving," and the like are to be understood to be open-ended,
i.e., to mean including but not limited to. Only the transitional
phrases "consisting of" and "consisting essentially of,"
respectively, shall be closed or semi-closed transitional
phrases.
[0187] All of the compositions and/or methods disclosed and claimed
herein can be made and executed without undue experimentation in
light of the present disclosure. While the compositions and methods
of this invention have been described in terms of preferred
embodiments, it will be apparent to those of skill in the art that
variations may be applied to the compositions and/or methods and in
the steps or in the sequence of steps of the method described
herein without departing from the concept, spirit and scope of the
invention. More specifically, it will be apparent that certain
agents which are both chemically and physiologically related may be
substituted for the agents described herein while the same or
similar results would be achieved. All such similar substitutes and
modifications apparent to those skilled in the art are deemed to be
within the spirit, scope and concept of the invention as defined by
the appended claims.
* * * * *
References