U.S. patent application number 11/139205 was filed with the patent office on 2005-09-29 for treatments for immune systems disorders.
This patent application is currently assigned to NutraCea. Invention is credited to Cherukuri, Reddy Sastry V., Cheruvanky, Rukmini, Mazhar, Mohammed, McPeak, Patricia.
Application Number | 20050214392 11/139205 |
Document ID | / |
Family ID | 26683357 |
Filed Date | 2005-09-29 |
United States Patent
Application |
20050214392 |
Kind Code |
A1 |
McPeak, Patricia ; et
al. |
September 29, 2005 |
Treatments for immune systems disorders
Abstract
This invention provides methods and formulations for treating
disorders of the immune system comprising administering a fortified
formulation comprising stabilized rice bran derivative and a
fortification agent. Preferred rice bran derivatives are rice bran
oil and the solubilized fraction of rice bran. Preferred
fortification agents are glucosamine derivative,
methylsulfonylmethane, yucca concentrate, and grape seed
extract.
Inventors: |
McPeak, Patricia; (El Dorado
Hills, CA) ; Cheruvanky, Rukmini; (Folsom, CA)
; Cherukuri, Reddy Sastry V.; (Folsom, CA) ;
Mazhar, Mohammed; (El Dorado Hills, CA) |
Correspondence
Address: |
TOWNSEND AND TOWNSEND AND CREW, LLP
TWO EMBARCADERO CENTER
EIGHTH FLOOR
SAN FRANCISCO
CA
94111-3834
US
|
Assignee: |
NutraCea
El Dorado Hills
CA
|
Family ID: |
26683357 |
Appl. No.: |
11/139205 |
Filed: |
May 26, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11139205 |
May 26, 2005 |
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10012270 |
Nov 6, 2001 |
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6902739 |
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60307588 |
Jul 23, 2001 |
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Current U.S.
Class: |
424/750 ;
424/766; 514/62 |
Current CPC
Class: |
A61K 31/7008 20130101;
A61K 36/899 20130101; A61P 29/00 20180101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 31/7008 20130101; A61K 31/10 20130101;
A61P 25/04 20180101; A61K 36/899 20130101; A61P 3/02 20180101; A61K
36/87 20130101; A61P 19/02 20180101; A61K 36/87 20130101; A61P
29/02 20180101; A61K 2300/00 20130101; A61K 2300/00 20130101; A61K
31/10 20130101; A61P 19/04 20180101; A61P 19/10 20180101; A61P
43/00 20180101 |
Class at
Publication: |
424/750 ;
424/766; 514/062 |
International
Class: |
A61K 035/78 |
Claims
1-47. (canceled)
48. A method for treating a disorder of the immune system in a
mammal, said method comprising: administering a fortified
formulation comprising a stabilized rice bran derivative and at
least one fortification agent, thereby treating said disorder in
said mammal.
49. The method of claim 48, wherein said stabilized rice bran
derivative is a member selected from the group consisting of rice
bran oil, enzyme-treated stabilized rice bran, a solubilized
fraction, and a mixture thereof.
50. The method of claim 48, wherein said fortification agent is a
member selected from the group consisting of a glucosamine
derivative, methylsulfonylmethane, yucca concentrate, and grape
seed extract.
51. A method for treating a disorder of the immune system in a
mammal, said method comprising: administering a fortified
formulation comprising a stabilized rice bran derivative and at
least one fortification agent, wherein said fortification agent is
a member selected from the group consisting of a glucosamine
derivative, methylsulfonylmethane, yucca concentrate, and grape
seed extract, wherein said stabilized rice bran derivative is a
member selected from the group consisting of rice bran oil,
enzyme-treated stabilized rice bran wherein the enzyme is
carbohydrate cleaving enzyme, a stabilized rice bran solubilized
fraction, and a mixture thereof, thereby treating said immune
system disorder in said mammal.
52. The method of claim 48 or 51, wherein said administering
comprises ingestion of said fortified formulation.
53. The method of claim 48 or 51, wherein said mammal is a member
selected from the group consisting of a human, a horse, a cat, and
a dog.
54. The method of claim 53, wherein said disorder of the immune
system is an autoimmune disease.
55. The method of claim 54, wherein said autoimmune disease is
selected from the group consisting of multiple sclerosis, type-1
diabetes mellitus, Graves' disease, systemic lupus erythematosus,
scleroderma, psoriasis, intestinal bowel disease and celiac
disease/gluten sensitivity.
56. The method of claim 55, wherein said mammal is a human.
57. The method of claim 55, wherein said disorder of the immune
system is intestinal bowel disease, and wherein said formulation is
ingested in the amount of about 2 grams to about 100 grams per day
total.
58. The method of claim 48 or 51, wherein said formulation is a
member selected from the group consisting of a cream, a capsule, a
bar, a powder, a food, a food supplement, a medical food, a liquid,
a beverage, an emulsion or mixture thereof.
Description
[0001] This application claims priority to U.S. Application Ser.
No. 60/307,588, filed Jul. 23, 2001, the disclosure of which is
incorporated herein by reference in its entirety for all
purposes.
FIELD OF THE INVENTION
[0002] This invention relates to fortified formulations and methods
for using such formulations for treating joint inflammation, pain,
and loss of mobility.
BACKGROUND OF THE INVENTION
[0003] Joint disorders and injuries are widespread, can cause
considerable discomfort, and cost billions of dollars in lost days
of work. Symptoms of these diseases and injuries include
inflammation, lameness, loss of mobility, and pain.
[0004] Arthritis is a multifactorial degenerative joint disease,
which progresses with age and results in joint stiffness,
inflammation, and pain. A joint is formed where two bones meet. The
healthy joint bones are lined with spongy cartilage which act as
shock absorbers, and the synovial fluid, which is secreted by the
synovial membrane lining the joint space, acts as a lubricant that
prevents friction. There are two major types of arthritis,
osteoarthritis, and rheumatoid arthritis.
[0005] Osteoarthritis is a condition occurring due to the
progressive degeneration and the wearing away of the cartilage (the
cushion between the joints), especially at the large joints like
the hips and knees. It is a normal age-related degenerative
process, occurring gradually after normal wear and tear. Being
overweight, inheriting the wrong genes, or simply growing old can
make the problem worse. Almost 80% of the people over the age of 60
years, all over the world, suffer from this disorder. OA begins
with the breakdown of cartilage resulting in pain, inflammation,
and progressive stiffness in the joint. Joint strengthening through
exercise, weight maintenance, and non-steroidal anti-inflammatory
drugs (NSAIDs) can alleviate symptoms. Osteoarthritis is often
accompanied by osteoporosis, a condition where the bone calcium
resorption occurs due to hormonal imbalance, making the bone more
brittle, which may lead to frequent fractures.
[0006] Rheumatoid arthritis is a prostaglandin-mediated joint
disease that leads to irreversible crippling of small joints,
especially the fingers and toes. This condition is difficult to
handle and only prostaglandin synthetase inhibitors can give some
relief to patients. Essentially, the body's immune system attacks
the cartilage, and the white blood cells (leukocytes) attack the
collagen. Statin drugs, which are immunomodulators, are used
frequently, though they have serious side effects. Temporary relief
may be obtained from NSAIDs. However, overuse of these drugs can
lead to ulcers.
[0007] Soft tissue rheumatism is a condition where many parts of
the body can be affected for a variety of reasons. In many
instances, soft tissue rheumatism is a sport-induced injury, such
as a sprain, tennis elbow, or runner's knee. Soft tissues are the
ligaments, tendons, and tendon sheath. Ligaments are bands of
tissue that connect bones. Tendons are bands of tissue that connect
muscle to bone. The tendon sheath is the tissue that surrounds and
lubricates the tendon. Injury to any of these soft tissues can
produce inflammation, pain, and stiffness. These conditions
typically clear up quickly, within several days to weeks, and are
usually treated with NSAIDs, icing the affected area, and rest.
Nevertheless, expedited recovery is still desirable, as these
injuries can cause considerable discomfort and reduce workplace
productivity.
[0008] In addition to the use of NSAIDs, the inflammatory response
can be regulated through the use multiple other drugs (see, Goodman
& Gilman's "The Pharmacological Basis of Therapeutics" eds.
Hardman et al. Ninth Edition, McGraw-Hill Publishing, 1996).
Unfortunately, certain anti-inflammatory drugs presently available
produce cytotoxic effects that reflect their initial employment as
cancer chemotherapeutics, typically anti-neoplastics. For example,
corticosteroids, which are often used for treatment of acute
inflammation, manifest significant adverse effects, such as
inducing Cushingoid features, skin thinning, increased
susceptibility to infection, and suppression of the
hypothalamic-pituitary-adrenal axis.
[0009] Joint injuries and conditions as described above also
afflict numerous other mammals, including domesticated animals such
as dogs, cats, and horses. In particular, horses often sustain
considerable joint injuries due to their participation in sporting
events or use for farm work. Lameness due to traumatic joint
disease is a common clinical problem in horses and is one of the
most important sources of financial losses in the equine
industry.
[0010] In recent years NSAIDS (non-steroidal anti-inflammatory
drugs), such as phenylbutazone, have been used to eliminate,
diminish, or at least assist in managing the lameness in
performance horses in all aspects of the horse industry (including
racing, cutting, reigning, hunter-jumper, dressage, rodeo,
barrel-racing). Unfortunately, NSAIDS require prescriptions and/or
veterinary dispensing, are costly, and are accompanied by mild
severe, and sometimes even catastrophic side effects.
[0011] As described previously, such methods for treatment of both
humans and animals only allow temporary relief and/or exhibit side
effects from prolonged use. Therefore, there is a need for safe and
effective treatment which can be used on a long-term basis without
side effects and which also promotes rebuilding of the
injured/diseased joints. The present invention fulfills this and
other needs.
BRIEF SUMMARY OF THE INVENTION
[0012] It has now been surprisingly found that a stabilized rice
bran derivative, having significant amounts of potent
phytonutrients and antioxidants, has an excellent effect in
relieving arthritic pain and pain associated with inflammation.
Fortification of these derivatives with certain minor herbal
components enhances their action. The fortified formulations
comprising a stabilized rice bran derivative and a fortification
agent reduce inflammation, pain, lameness, and loss of mobility.
These fortified formulations are more effective, have more
immediate action, and require lower dosages than currently existing
formulations for such conditions.
[0013] As such, the present invention provides a method for
treating an inflammatory disease or reducing an inflammatory
reaction in a mammal comprising administering a stabilized rice
bran derivative and a fortification agent. The stabilized rice bran
derivative can include, but is not limited to rice bran oil,
enzyme-treated stabilized rice bran, a solubilized fraction, or
mixtures thereof. Preferably, the stabilized rice bran derivative
is rice bran oil or the solubilized fraction. The fortification
agent can include, but is not limited to, a glucosamine derivative,
methylsulfonylmethane, yucca concentrate, or grape seed extract. In
preferred embodiments, the administering comprises ingestion of the
formulation and the inflammatory disease is a disorder of the bone
joint, including, but not limited to, osteoarthritis, osteoporosis,
rheumatoid arthritis, and soft tissue rheumatism. The mammal is
typically a human or a horse.
[0014] In another aspect, the present invention provides a method
for treating lameness or loss of mobility in a mammal comprising
administration of a formulation comprising a stabilized rice bran
derivative and a fortification agent.
[0015] In yet another aspect, the present invention provides a
method for reducing pain in a mammal, the method comprising
administration of a formulation comprising a stabilized rice bran
derivative and a fortification agent.
[0016] In still yet another aspect, the present invention provides
a method for reducing prostaglandin synthetase activity, the method
comprising administration of a formulation comprising a stabilized
rice bran derivative and a fortification agent.
[0017] In another aspect, the invention provides a formulation
comprising a stabilized rice bran derivative and fortification
agent for treating joint inflammation and loss of mobility. In
certain embodiments, the fortification agent is a glucosamine
derivative, methylsulfonylmethane, yucca concentrate, or grape seed
extract.
[0018] Also provided are formulations and methods for treatment of
an inflammatory disease, lameness, and loss of mobility comprising
administering a tocol composition.
[0019] These and other aspects will become more apparent when read
with the detailed description and examples, which follow.
DEFINITIONS
[0020] Unless otherwise specified, the following terms used in the
specification and claims have the meanings given below.
[0021] The term "fortification agent" refers to any agent which
improves the ability of the stabilized rice bran derivative to
treat a pathological condition of the joint, such as an
inflammatory disease, lameness, loss of mobility, pain. etc.
Preferred fortification agents include glucosamine derivatives,
methylsulfonylmethane, yucca concentrate, grape seed extract and
combinations thereof.
[0022] The term "glucosamine derivative" refers to modified
versions of glucosamine, such as glucosamine sulfate and N-acetyl
glucosamine. These derivatives are the building blocks of
proteoglycans, which are used in cartilage synthesis.
[0023] The term "methylsulfonylmethane" (MSM) refers to a chemical
compound that is a sulfur donor. Depletion of sulfur amino acids
leads to arthritis. Sulfur compounds aid in the synthesis of
proteoglycans and glucosaminoglycans, which form the basic matrix
of cartilage.
[0024] The term "yucca concentrate" refers to a preparation of the
root of the yucca plant. Yucca is a desert plant that is rich in a
steroidal saponin (sarasaponin). This saponin helps the body's
production of cortisone and improves the ratio of
cortisol/DHEA.
[0025] The term "grape seed extract" refers to an extract from
grape seeds which has antioxidant effects.
[0026] The term "administering" refers to the manner in which a
therapeutic agent is introduced to a mammal with a particular
condition. Such administration may be by any one of the various
standard routes for administration of drugs, i.e., topical, oral
(ingestion by the mammal), parenteral, etc.
[0027] The term "reducing lameness or loss of mobility" refers to
improving the function of any joint. Joint function can be measured
by evaluating parameters such as the range of motion and presence
of discomfort or pain during movement.
[0028] The term "reducing pain in or around a bone joint" refers to
alleviating pain localized around a bone joint.
[0029] The term "prostaglandin synthetase" (also termed
prostaglandin endoperoxide synthase) refers to the cyclooxygenase
(COX) enzymes which catalyze the conversion of arachidonic acid to
prostanoids.
[0030] The term "disorder of the bone joint" refers to any
condition where the normal function of the joint, i.e., range of
motion, ability to bear weight and the like is impaired, or where
use of the joint is accompanied by discomfort or pain. These
conditions include those where impairment of the joint is the
primary symptom or ones where the disorder has multiple symptoms.
These disorders typically affect the bone, joint capsule,
cartilage, tendons, ligaments, tendon sheaths, bursa, synovial
fluid, etc. Common disorders of the joint include osteoarthritis,
rheumatoid arthritis, gout, lupus, tendonitis, bursitis, carpal
tunnel syndrome, sprains, etc.
[0031] The term "osteoarthritis" refers to a degenerative joint
disease where the cartilage that normally cushions the joint and
protects it from impact erodes.
[0032] The term "rheumatoid arthritis" refers to an autoimmune
disease with inflammation of the lining of the joints (synovial
membrane). The thickened synovial membrane can erode the
surrounding ligaments and bone.
[0033] The term "soft tissue rheumatism" refers to conditions where
the soft tissues of the body, such as ligaments, tendons, and the
tendon sheath are injured.
[0034] The term "tocol" refers to E complex vitamins known as
tocopherols and tocotrienols which have antioxidant properties.
There are at least ten different isomeric forms of these vitamins.
The term "tocol composition" refers to any composition comprising
tocols.
[0035] As used herein the term "enzyme treated stabilized rice bran
derivative" refers to an enzyme-treated stabilized rice bran made
by mixing a stabilized rice bran with an aqueous solution in a 15%
to about a 35% aqueous slurry w/w; adding an enzyme to the aqueous
rice bran slurry to convert starch to dextrin, and then directly
drying the dextrin solution to form an enzyme treated stabilized
rice bran derivative. The enzyme treated stabilized rice bran
comprises about 20% to about 30% total dietary fiber.
[0036] As used herein the term "GRAS" means generally regarded as
safe with respect to food additives.
[0037] As used herein the term "stabilized rice bran derivative
solubilized fraction" refers to a fraction during a partitioning
process. Specifically, after the stabilized rice bran aqueous
slurry is enzymatically treated, it is then pumped into a
centrifuge where the insoluble fraction precipitates out of the
aqueous solution. The aqueous material is pumped to a dryer and
then dried. This dried aqueous portion produces the soluble
fraction. The constituent parts and their percentages are listed in
the Tables below.
BRIEF DESCRIPTION OF THE DRAWINGS
[0038] FIG. 1 illustrates the standards used to score the degree of
lameness of horses. Horses are evaluated for extension, toe
placement, and flexion.
[0039] FIG. 2 illustrates the effect of formulations used in
methods of this invention versus other commercially available
treatments/NSAIDs on lameness of horses. The graph measures
soundness score (measurement of lameness) after treatment with
"bute", Cosequin, Absorbine Flex over 50 days.
[0040] FIG. 3 illustrates the effect of treatment with formulations
of this invention versus other commercially available
nutraceuticals/NSAIDs on levels of carrageenan-induced inflammation
on Day 21 of the clinical trial. Injection is induced and then the
size of inflammation at the site of injection with carrageenan is
measured over 7 hours for horses treated with the negative control,
"bute", Cosequin, Next Level, or Absorbine Flex.
[0041] FIG. 4 illustrates the effect of treatment with formulations
of this invention versus other commercially available
nutraceuticals/NSAIDs on levels of carrageenan-induced inflammation
on Day 50 of the clinical trial.
DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED EMBODIMENTS
I. COMPOUNDS USED IN METHODS AND FORMULATIONS OF THIS INVENTION
[0042] Compounds used in formulations and methods of this invention
include stabilized rice bran derivatives, which can include, but
are not limited to rice bran oil, enzyme-treated stabilized rice
bran, a solubilized fraction, or mixtures thereof. Preferably, the
stabilized rice bran derivative is rice bran oil or the solubilized
fraction. Other compounds used in formulations and methods of this
invention include fortification agents which can include, but are
not limited to, a glucosamine derivative, methylsulfonylmethane,
yucca concentrate, grape seed extract, curcumin, ginger powder,
boswellin, aswagandha, and hempseed oil. The compounds of the
invention may also comprise an extract of active ingredients of
rice bran derivatives, such as tocols.
[0043] A. Stabilized Rice Bran Derivatives
[0044] In harvested rice, also known as rough rice, the kernel is
completely enveloped by the rice hull. The milling process removes
the hull, which yields brown rice. The outer brown layer is then
removed by an abrasive milling process to generate white rice. The
separated brown layer is designated rice bran.
[0045] Rice bran is the mesocarp, i.e., the portion between the
hull and rice grain, obtained by milling or polishing brown rice.
It constitutes about 10% of rough rice. It is generally used as an
animal feed. It contains about 18-24% fat, about 25% dietary fiber,
about 14% protein and about 45% total carbohydrates besides several
potent micronutrients. It is rich in B-complex vitamins, vitamin E
and its isomers, minerals like potassium, magnesium, and
phosphorous besides several potent antioxidants.
[0046] Stabilized rice bran can be commercially purchased or
prepared using various methods. Most stabilization methods of rice
bran result in inactivation of the lipases which are present,
inactivation of the peroxidases, and inactivation of the
microorganisms, while still maintaining the high levels of
antioxidants in the rice bran. For a general discussion of
stabilization and processing see, Rice Science and Technology,
edited by W. E. Marshall and James I Wadswoth, (1994) pages
390-404.
[0047] Stabilized rice bran is available commercially from
Producers Rice Mill Inc. (Stuttgart, Ark.), Riceland Foods
(Stuttgart, Ark.), Riviana Foods, Inc. (Houston, Tex.), Uncle Ben's
Inc. (Houston, Tex.) and The RiceX.TM. Company (El Dorado Hills,
Calif.). Due to different stabilization processes, stabilized rice
bran will differ in composition and stabilization characteristics
when derived from different manufacturers.
[0048] In order to generate the rice bran derivatives for use in
the present invention, the rice bran is first stabilized, and then
it is further separated into at least two fractions. These include,
but are not limited to, a stabilized rice bran soluble derivative
and a stabilized rice bran insoluble derivative. Preferably, the
separation into the rice bran derivatives includes a nonchemical
process i.e., an enzymatic process. In this process, partitioning
or fractionation preferably proceeds as outlined hereinafter.
[0049] The stabilized rice bran is made into about a 15% to about a
35% slurry, preferably, 20-25% slurry with potable water. An
enzyme, which can include, but is not limited to, a dextranase, a
maltase, an a-amylase, and various other carbohydrate cleaving
enzymes, is added to the batch converting the starch to dextrins.
The slurry is heated to about 150.degree. F. to about 200.degree.
F. using for instance, a steam injection cooker, a heat exchanger,
or other heating method. The slurry is then pumped to a horizontal
centrifuge wherein the insoluble fraction is separated. The
insoluble fraction is collected and then dried on a belt dryer, and
subsequently ground into a powder. This powder is the stabilized
rice bran insoluble fraction. The aqueous material is pumped to a
drum dryer and then dried. This dried aqueous portion produces the
stabilized rice bran solubilized fraction.
[0050] The enzyme-treated stabilized rice bran can be generated
using the rice bran slurry as described above. The process for
making an enzyme-treated stabilized rice bran derivative can
comprise admixing stabilized rice bran with an aqueous solution to
form about a 15% to about a 35% aqueous rice bran slurry,
preferably a 20% to about a 30% aqueous rice bran slurry w/w;
adding an enzyme to the aqueous rice bran slurry to convert starch
to dextrin, thereby forming an enzyme-treated slurry, and then
directly drying the enzyme-treated slurry to form an enzyme-treated
stabilized rice bran derivative.
[0051] Preferably, after the enzyme is added to the slurry, the
slurry is heated to about 150.degree. F. to about 200.degree. F.
The slurry is then dried, wherein the drying is accomplished by a
process such as belt drying, spray drying, drum drying and
air-drying.
[0052] These stabilized rice bran derivatives are also available
commercially from NutraStar and the RiceX.TM. Company of
California. For the purpose of the invention, stabilized rice bran
is available from NutraStar as StaBran.TM. or from RiceX.TM. as
Stabilized Rice Bran. The soluble derivative is available from
NutraStar as RiSolubles.TM., non-chemically predigested and
separated from insoluble fiber, or RiceX.TM. Ricelin.TM. from the
RiceX.TM. Company. NutraStar and the RiceX.TM. Company are located
in El Dorado Hills, Calif. Rice bran oil is extracted from rice
bran using standard methods known in the art for extracting oils
such as peanut oil. The insoluble derivative is available as
RiceX.TM. Fiber Concentrate.
[0053] These derivatives have been shown to have more than a
hundred (100) potent anti-oxidants. The major antioxidant vitamin E
and its isomers known as tocopherols (T) and tocotrienols (T.sub.3)
are collectively called tocols. A tocol-rich substance is a mixture
containing one or more compounds selected from tocopherols (T),
tocotrienols (T.sub.3), and tocotrienol-like (T.sub.3-like)
compounds. Stabilized rice bran is the highest natural source of
vitamin E.
[0054] Antioxidants in stabilized rice bran derivatives include,
but are not limited to, .gamma.-oryzanol, .beta.-carotene, several
known flavanoids, phytosterols, lipoic acid, and ferulic acid. Some
of these compounds are present at a high concentration, much more
than in any of the known natural sources.
[0055] The processing of rice bran and the nutritional composition
of rice bran, as well as other aspects of the stabilized rice bran
derivatives used in formulations of this invention, are further
described in issued U.S. Pat. No. 6,126,943, entitled "Method for
Treating Hypercholesterolemia, Hyperlipidemia, and
Atherosclerosis", and allowed U.S. patent application Ser. No.
09/624,474, which are both incorporated herein by reference.
[0056] Table I below sets forth the NutraStar StaBran.TM. Product
Data Sheet comprising stabilized Rice Bran and Germ, non-chemically
treated to deactivate lipase.
1TABLE I NutraStar StaBran .TM. Regular Product Data Sheet
INGREDIENTS: Stabilized Rice Bran and Germ, non-chemically treated
to deactivate Lipase and ensure total stability. GUARANTEED
SPECIFICATIONS: Protein 12-16% Soluble 2-6% Fiber Fat 18-23% Ash
7-10% Total 45-55% Moisture 4-8% Carbohydrates Total Dietary 23-35%
Free Fatty <3% Fiber Acids MICROBIOLOGICAL: Total Plate Maximum
10,000 CFU/g. Count Total Maximum 100 CFU/g. Coliform E. coli
Maximum <10 CFU/g. Salmonella Negative Yeast Maximum 100 CFU/g.
Mold Maximum 100 CFU/g. PHYSICAL: Appearance Granular Solid Color
Light brown/tan .paragraph. Flavor Nutty, Toasted Bulk Density 0.47
(g/cc) ANALYTICAL DATA MACRONUTRIENTS (g/100 g) Protein (N .times.
6.25) 14.50 Fat 20.50 Saturated Fatty Acids 3.70 Total Carbohydrate
51.00 Available Carbohydrate 22.00 Ash 8.00 Moisture (100 degree
vac.) 6.00 Crude Fiber 7.30 Total Dietary Fiber 29.00 Soluble Fiber
2.00 Calories/100 g. 330.50 VITAMINS Vitamin A; Carotenoids
(mcg/100 g) .beta.-Carotene 37.00 .alpha.-Carotene 0.40 Lycopene
2.30 Lutein 63.80 Zeaxanthin 18.40 Precryptoxanthin/Cryptoxanthin
7.40 Total Carotenoids 129.30 Vitamin B Complex (mg/100 g) Vitamin
B1 2.70 Vitamin B2 0.28 Vitamin B3 46.90 Vitamin B5 3.98 Vitamin B6
3.17 Vitamin B12 (mcg/100 g) <0.500 Vitamin C(mg/100 g)
<0.500 Vitamin E Complex (mg/100 g) Tocopherols (T) 12.00
Tocotrienols (T3) 13.60 Total Tocols (T + T3) 25.60 Other
Micronutrients (mg/100 g) Folic Acid (mcg/100 g) 26.60 Biotin
(mcg/100 g) 14.10 Choline 104.80 Inositol 1496.0 .gamma.-Oryzanol
245.15 Phytosterols (mg/100 g) .beta.-Sitosterol 167.67
Stigmasterol 62.64 Campesterol 96.23 Brassicasterol 14.61 Total
Phytosterols 341.15 MINERALS (mg/100 g) Sodium 8.00 Potassium
1573.00 Calcium 40.00 Magnesium 727.00 Phosphorous 1591.00
Manganese 25.60 Iron 7.70 Copper 0.27 Zinc 5.50 Chromium (ppm)
<1 ppm Total Sugars(g/100 g) 8.09 (No Lactose)
[0057] Table II below sets forth NutraStar RiSolubles.TM. product
data sheet that comprises stabilized Rice Bran and Germ,
non-chemically predigested and separated from insoluble fiber.
2TABLE II NutraStar RiSolubles .TM. Product Data Sheet INGREDIENTS:
Stabilized Rice Bran and Germ, non-chemically predigested and
separated from insoluble fiber. GUARANTEED SPECIFICATIONS: Protein
7-12% Ash 3-7% Fat 25-32% Moisture 2-7% Total Carbohydrates 50-60%
Free Fatty Acids <3% Total Dietary Fiber 0-6% MICROBIOLOGICAL:
Total Plate Count Maximum 10,000 CFU/g. Total Coliform Maximum 100
CFU/g. E. coli Maximum <10 CFU/g. Salmonella Negative Yeast
Maximum 100 CFU/g. Mold Maximum 100 CFU/g. PHYSICAL: Appearance
Fine Powder Color Pale Yellow Flavor Sweet, Nutty Bulk Density
(G/Cc) 0.31 ANALYTICAL DATA MACRONUTRIENTS (g/100 g) Protein (N
.times. 6.25) 7.50 Fat 26.50 Saturated Fatty Acid 4.80 Total
Carbohydrate 57.50 Available Carbohydrate 54.50 Ash 5.00 Moisture
(100 degree vac.) 3.00 Crude Fiber 4.60 Total Dietary Fiber 3.00
Soluble Fiber 3.00 Calories/100 g. 486.50 VITAMINS Vitamin A;
Carotenoids (mcg/100 g) .beta.-Carotene 8.10 .alpha.-Carotene 0.00
Lycopene 0.20 Lutein 26.10 Zeaxanthin 10.90
Precryptoxanthin/Cryptoxanthin 1.27 Total Carotenoids 46.57 Vitamin
B Complex (mg/100 g) Vitamin B1 3.60 Vitamin B2 0.46 Vitamin B3
76.60 Vitamin B5 5.82 Vitanin B6 5.81 Vitamin B12 (mcg/100 g)
<0.500 Vitamin C (mg/100 g) <0.500 Vitamin E Complex (mg/100
g) Tocopherols (T) 8.00 Tocotrienols (T3) 10.00 Total Tocols (T +
T3) 18.00 Other Micronutrients (mg/100 g) Folic Acid (mcg/100 g)
36.17 Biotin (mcg/100 g) 14.70 Choline 150.00 Inositol 1490.0
.gamma.-Oryzanol 248.10 Phytosterols (mg/100 g) .beta.-Sitosterol
211.90 Stigmasterol 68.69 Campesterol 117.32 Brassicasterol 15.25
Total Phytosterols 413.16 MINERALS (mg/100 g) Sodium 15.75
Potassium 1562.00 Calcium 8.30 Magnesium 170.80 Phosphorous 763.00
Manganese 3.20 Iron 1.90 Copper 0.07 Zinc 1.75 Chromium (ppm) <1
ppm Total Sugars (g/100 g) 13.83 (No Lactose)
[0058] Table III below sets forth the product data sheet for Rice
bran oil.
3TABLE III Rice Bran Oil Refined Product Data Sheet SPECIFICATIONS
Physicochemical Parameters: Appearance Clear Color (51/4" Lovibond
Red) 3.5 max. Moisture (%) 0.05-0.10 Specific gravity @ 25.degree.
C. 0.910-0.920 Refractive Index @ 40.degree. C. 1.460-1.470 Iodine
Value 90-105 Saponification Value 180-190 Acid Value <0.5
Unsaponifiables <3% Smoke Point >213.degree. C./415.degree.
F. Flash Point >250.degree. C./480.degree. F. Fatty Acid Profile
(%): Myristic Acid (C14:0) 0.3-0.5 Palmitic Acid (C16:0) 14.0-20.0
Stearic Acid (C18:0) 1.2-2.0 Oleic Acid (C18:1) 40-44 Linoleic
(C18:2) 34-40 Linolenic (C18:3) 1.0-2.0 Micronutrients: Tocopherols
& Tocotrienols 500-600 ppm ANALYSIS Physicochemical Parameters:
Appearance Clear Color (51/4" Lovibond red) 3.5 max Specific
gravity @ 25.degree. C. 0.916 Refractive Index @ 40.degree. C. 1.47
Smoke Point 213.degree. C. Iodine Value 104 Saponification Value
187 Acid Value <0.5 Unsaponifiables <3% Fatty Acid Profile
(%): Pahnitic Acid (C 16:0) 16.0 Stearic Acid (C18:0) 2.0 Oleic
Acid (C18:1) 42.0 Linoleic (C18:2) 38.0 Linolenic (C18:3) 2.0
Micronutrients: Tocopherols & Tocotrienols 600 ppm
[0059]
4TABLE IV Antioxidants in NutraStar's Rice Bran Products Gamma
Oryzanol (2200-3000 ppm) Gamma Oryzanol is not a single component.
It is a mixture of 20 components having different antioxidant
properties. Cycloartenol trans-ferulate Cycloartenol cis-ferulate
Cycloartanol trans-ferulate Cycloartanol cis-ferulate
Cycloeucalenol trans-ferulate Cycloeucalenol cis-ferulate
24-Methylenecycloartanol trans-ferulate 24-Methylenecycloartanol
cis-ferulate 24-Methylcholesterol trans-ferulate
24-Methylcholesterol cis-ferulate .beta.-Sitosterol trans-ferulate
.beta.-Sitosterol cis-ferulate .beta.-Sitostenol trans-ferulate
.beta.-Sitostenol cis-ferulate Stigmasterol trans-ferulate
Stigmasterol cis-ferulate Stigmastenol trans-ferulate Stigmastenol
cis-ferulate Campesterol trans-ferulate Campesterol cis-ferulate
Tocopherols & Tocotrienols (220-320 ppm) Tocopherols and
tocotrienols belong to the same chemical group, but exist in 10
different isomeric forms having different antioxidant properties.
.alpha.-Tocopherol .beta.-Tocopherol .gamma.-Tocopherol
.delta.-Tocopherol .alpha.-Tocotrienol .beta.-Tocotrienol
.gamma.-Tocotrienol .delta.-Tocotrienol Desmethyl-tocotrienol
Didesmethyl tocotrienol Polyphenols Ferulic acid .alpha.-Lipoic
acid Methyl ferulate .rho.-Coumaric acid .rho.-Sinapic acid
Isovitexin Proanthocyanidins Metal Chelators Magnesium (6250-8440)
Calcium (303-500) Phosphorous (14,700-17,000) Phytosterols (21
Components) (2230-4400 ppm) .beta.-Sitosterol Campesterol
Stigmasterol Sitostenol .DELTA..sup.5-Avinasterol
.DELTA..sup.7-Stigmastenol Sterol glucoside Acylsterol glucoside
Oligoglycosylsterol Monoglycosylsterol Cellotetraosylsitosterol
Methylsterol Dimethylsterol Gramisterol Isofucosterol Obtusifoliol
Branosterol 28-Homotyphasterol 28-Homosteasteronic acids
6-Deoxycastasterone .beta.-Amyrin Carotenoids (0.9-1.6 ppm)
.alpha.-Carotene .beta.-Carotene Lycopene Lutein Zeaxanthine Amino
Acids Tryptophan (2100) Histidine (3800) Methionine (2500) Cystein
(336-448) Cystine (336-448) Arginine (10800) B Vitamins Thiamin
(22-31) Riboflavin (2.5-3.5) Niacin (370-660) Pantothenic acid
(36-50) Pyridoxine (29-42) Betaine Dimethyl glycine Inositol
(12000-18,800) Biotin (0.1-2.2) Choline (930-1150) Folic acid
(0.20-0.30) Phytates (1500-1750) Polysaccharides
Cycloartenol-ferulic acid glycoside Diferulic acid complex
Diferulic acid-calcium complex Hemicelluloses Arabinogalactan
Arabinoxylan Xyloglucan Proteoglycan Glycoprotein Arabinofuranoside
Phosphollpids Phosphatidylserine PhosphatidylCholine
Phosphatidylethanolamine Lysophophatidylcholine
Lysophosphatidylethanolamine Enzymes Glutathione peroxidase
Methionine reductase Superoxide dismutase Polyphenol oxidase
Catalase Coenzyme Q10 Aspartate amino transferase isozyme AAT-1
& AAT-2
[0060] B. Fortification Agents
[0061] Other compounds used in formulations and methods of this
invention include fortification agents which can include, but are
not limited to, a glucosamine derivative, methylsulfonylmethane,
yucca concentrate, grape seed extract, curcumin, ginger powder,
boswellin, aswagandha, and hempseed oil.
[0062] Glucosamine sulfate and N-acetylglucosamine can be extracted
from crustacean shells. Yucca concentrate is derived from the root
of the yucca plant. Methylsulfonylmethane, curcumin, ginger, and
boswellin, aswagandha, and hempseed oil are derived from plant
sources.
[0063] C. Extracts ofActive Ingredients of Stabilized Rice Bran
[0064] In certain aspects, the active ingredients described above
can also be extracted from the stabilized rice bran derivatives and
used individually to treating inflammation, pain, lameness, and
loss of mobility.
II. ACTIVITY OF COMPOUNDS USED IN THE METHODS AND FORMULATIONS
[0065] The present invention is based on the discovery that rice
bran derivatives and extracts of their active ingredients are
useful for treating inflammatory reactions, pain, lameness, and
loss of mobility. These components seem to act synergistically,
creating an enhanced effect not expected when one evaluates the
individual compounds present in rice bran derivatives. It is
believed that the synergestic activity arises between dense
phytonutrients and antioxidants present in the rice bran
derivatives.
[0066] Without being bound by any particular theory, it is believed
that the mechanisms of action of the individual bioactive
components in the stability of rice bran derivatives include, but
are not limited to, the following:
[0067] 1. Phytosterols: Beta-sitosterol and its glycosides improve
immune function.
[0068] 2. Gamma-oryzanol acts as an antioxidant and
anti-inflammatory agent.
[0069] 3. Tocotrienols inhibit prostaglandin synthetase activity,
which reduces the inflammatory response by suppressing the
pro-inflammatory cytokines (IL-4) and elevating the
anti-inflammatory cytokines (IL-2).
[0070] 4. Tocopherols act as antioxidants.
[0071] 5. Minerals such as magnesium and other trace materials
increase absorption of calcium and help in healthy bone
mineralization.
[0072] 6. Proteoglycans, the matrix of collagen, are synthesized
from the amino acid pool and polysaccharide units from the rice
bran derivative.
[0073] 7. Polyphenols such as ferulic acid, tocopherols and
gamma-oryzanol present in rice bran products are potent
anti-oxidants which help in joint function.
[0074] 8. Omega-3 fatty acids in rice bran products enhance the
anti-inflammatory cytokines and inhibit pro-inflammatory cytokines.
They are also powerful COX-2 inhibitors.
[0075] The invention is further based on the discovery that the
administration of stabilized rice bran derivatives with other
bioactive compounds are particularly useful for treating
inflammatory reactions, pain, lameness, and loss of mobility. Such
fortified formulations are more effective, have more immediate
action, and require lower dosages than currently existing
formulations for such conditions. In certain instances, these
components act synergistically, creating an enhanced effect which
is greater than the individual compounds acting alone or
additively. This synergy is completely unexpected when one
evaluates the individual bioactive compounds present in the
formulation.
[0076] Without being bound by any particular theory, it is believed
that the mechanisms of action of these components include, but are
not limited to, the following:
[0077] 1. Yucca concentrate is rich in steroidal saponins, which
increases cortisone production. Cortisone has an anti-inflammatory
effect.
[0078] 2. Glucosamine derivatives, such as n-acetyl glucosamine and
glucosamine sulfate, promote repair of damaged connective tissues.
Specifically, these derivatives promote cartilage production.
[0079] 3. Methylsulfonylmethane is a sulfur donor for the synthesis
of collagen. It maintains the synovial fluid and facilitates the
absorption of vitamin C, biotin, and panthothenic acid.
[0080] 4. Grape seed extract is a potent antioxidant that helps
prevent free radical damage at the joints.
[0081] 5. Curcumin and ginger powder have anti-inflammatory
effects.
[0082] 6. Boswellin is a prostaglandin synthetase inhibitor.
[0083] 7. Aswagandha is a COX-2 inhibitor.
[0084] 7. Hempseed oil is rich in omega-3 fatty acids, which have
antiinflammatory effects.
[0085] Although the mechanism of action of the compounds of the
present invention discussed above are believed to be correct, it
should in no way be considered as limiting the present invention.
Those of skill in the art will understand that the various
embodiments of the invention may be practiced regardless of the
model used to describe the theoretical underpinnings of the
invention.
[0086] Animal models that are widely viewed to reflect inflammatory
responses and to have predictive value in assessing the efficacy of
various treatments for these disorders can be utilized to evaluate
the therapeutic efficacy of the compounds described herein. As
described in Example 1, the effects of the compounds on joint
inflammation and lameness can be assessed in horses with
trauma-induced inflammation. Alternatively, improvement of
inflammation and mobility can be measured with a collagen-induced
rheumatoid arthritis mouse model (see, Enokida M et aL (2001) Bone
28(1): 87-93)
III. FORMULATION AND DOSAGES USED IN METHODS OF THIS INVENTION
[0087] The present invention also provides various formulations of
stabilized rice bran derivatives and their extracted active
ingredients with fortification agents. These formulations include
both neutraceutical formulations and standard pharmaceutical
compositions.
[0088] A. Nutraceuticals
[0089] The nutraceutical formulations of this invention can take a
variety of forms, such as a powder, a food, a food supplement, a
medical food, a liquid, a beverage, an emulsion or mixtures
thereof.
[0090] To incorporate the fortified formulation of the rice bran
derivative into the diet of a mammal various options include, but
are not limited to, simply sprinkling the formulation on another
food substance (i.e., salad, bread, cereal, etc.), being a major
ingredient in a multigrain ready-to-eat cereal, incorporating it
into a baked product (breads, muffins, waffles, etc.), pasta,
healthy dessert and snacks (athletic bar, healthy drink, etc.) and
high fiber foods.
[0091] For administration to humans, the fortified formulation of
rice bran derivative is preferably a drink, a capsule, or a bar.
These formulations typically comprise the solubilized fraction of
stabilized rice bran or rice bran oil. Such formulations further
comprise a fortification agent which promotes joint health,
including, but not limited to, glucosamine derivatives (such as
n-acetyl glucosamine, glucosamine sulfate), methylsulfonylmethane,
yucca concentrate, and grape seed extract. Additives to improve
flavor and consistency of the product are also included in the
formulation. Especially preferred formulations and dosages are
described in Example 2.
[0092] For administration to animals, the fortified formulation of
rice bran derivative is a powder or liquid. These formulations
typically comprise the solubilized fraction of rice bran or rice
bran oil. Such formulations also comprise a fortification agent,
including, but not limited to, glucosamine derivatives (such as
n-acetyl glucosamine, glucosamine sulfate), methylsulfonylmethane,
and yucca concentrate. Additives to improve flavor and consistency
of the product are also included in the formulation. Especially
preferred formulations and dosages are described in Example 2.
[0093] B. Pharmaceutical Compositions
[0094] In other preferred embodiments, formulations of this
invention are pharmaceutical compositions suitable for
administration via various routes, preferably orally or topically,
and for therapeutic and/or prophylactic administration. A number of
suitable formulations for use in the present invention are found in
Remington's Pharmaceutical Sciences (Mack Publishing Company,
Philadelphia, Pa., 17th ed.,1985) and in Dermatological
Formulations: Percutaneous absorption, Barry (Ed.), Marcel Dekker
Inc., 1983, both incorporated herein by reference. Moreover, for a
brief review of methods for drug delivery, see Langer, Science
249:1527-1533, 1990, which is also incorporated herein by
reference. The pharmaceutical compositions described herein can be
manufactured in a manner that is known to those of skill in the
art, i.e., by means of conventional mixing, dissolving, levigating,
emulsifying, encapsulating, entrapping or lyophilizing processes.
It will be appreciated that the present methods and excipients are
merely exemplary and are in no way limiting.
[0095] 1. Topical Administration
[0096] More particularly, these compounds can be formulated into
preparations in solid, semi-solid, or liquid forms suitable for
local or topical administration, such as gels, water soluble (e.g.,
K-Y) jellies, creams, lotions, suspensions, foams, powders,
slurries, ointments, solutions, oils, pastes, suppositories,
sprays, emulsions, saline solutions, dimethylsulfoxide (DMSO)-based
solutions. In general, carriers with higher densities, such as K-Y
jelly, are capable of providing an area with a prolonged exposure
to the active ingredients. In contrast, a solution formulation
provides more immediate exposure of the active ingredient to the
chosen area, although the effects generally do not last as
long.
[0097] The pharmaceutical compositions also may comprise suitable
solid or gel phase carriers or excipients, which are compounds that
allow increased penetration of, or assist in the delivery of,
therapeutic molecules across the stratum comeum permeability
barrier of the skin. There are many of these penetration-enhancing
molecules known to those trained in the art of topical formulation.
Examples of such carriers and excipients include, but are not
limited to, humectants (e.g., urea), glycols (e.g., propylene
glycol), alcohols (e.g., ethanol), fatty acids (e.g., oleic acid),
surfactants (e.g., isopropyl myristate and sodium lauryl sulfate),
pyrrolidones, glycerol monolaurate, sulfoxides, terpenes (e.g.,
menthol), amines, amides, alkanes, alkanols, ORGELASE, water,
calcium carbonate, calcium phosphate, various sugars, starches,
cellulose derivatives, gelatin, and polymers such as polyethylene
glycols.
[0098] 2. Oral Administration
[0099] For enteral administration the compounds useful in the
methods of the invention can be administered in either single or
multiple dosages. The compounds may be administered in combination
with pharmaceutically acceptable carriers in a variety of dosage
forms. For example, capsules, lozenges, hard candies, powders,
sprays, aqueous suspensions, elixirs, syrups, and the like may be
formulated with various pharmaceutically acceptable inert carriers.
Such carriers include solid diluents or fillers, sterile aqueous
media and various non-toxic organic solvents.
[0100] Tablets may contain various excipients such as sodium
citrate, calcium carbonate and calcium phosphate, along with
various disintegrants such as starch (preferably potato or tapioca
starch), alginic acid and certain complex silicates, together with
binding agents such as polyvinylpyrrolidone, sucrose, gelatin and
acacia. Additionally, lubricating agents such as magnesium
stearate, sodium lauryl sulfate and talc are often very useful for
tabletting purposes. Solid compositions of a similar type may also
be employed as fillers in soft and hard-filled gelatin capsules;
preferred materials in this connection would also include lactose
or milk sugar as well as high molecular weight polyethylene
glycols. When aqueous suspensions and/or elixirs are desired for
oral administration, the essential active ingredient therein may be
combined with various sweetening or flavoring agents, coloring
matter or dyes and, if so desired, emulsifying and/or suspending
agents, together with such diluents as water, ethanol, propylene
glycol, glycerin and various combinations thereof.
[0101] 3. Dosages
[0102] Pharmaceutical compositions suitable for use in the present
methods include compositions wherein the active ingredients are
contained in a therapeutically or prophylactically effective
amount. The amount of compound or composition administered will, of
course, be dependent on the subject being treated, on the subject's
weight, the severity of the affliction, and the manner of
administration. A typical dosage for enteral administration is an
amount from about 2 grams to about 100 grams per day. Determination
of an effective amount is well within the capability of those
skilled in the art, especially in light of the disclosure provided
supra.
[0103] As used herein, "effective amount," or "therapeutically
effective amount" refers to an amount of any of the compounds or
formulations used in methods of the present invention that results
in treatment of the medical condition, i.e., reduction in pain,
redness, inflammation, lameness, or any other symptom.
Alternatively, an "effective amount" may be determined by
monitoring reduction in any detectable symptom of the condition,
such as the degree of swelling, inflammation, redness, size of the
affected area, range of motion, and the like. In the context of the
present invention, "prophylactically effective amount" refers to an
amount of any of the present compounds that prevents the
development or relapse of a medical condition. For example, a
"prophylactically effective amount" is an amount that protects a
subject from developing an inflammatory disorder of the joint.
[0104] For any compound or formulation used in a method of the
invention, a therapeutically effective dose can be estimated
initially from animal models (described supra), well known to those
of skill in the art. Such information can be used to more
accurately determine useful doses in humans. Initial dosages can
also be estimated from in vitro or in vivo data.
[0105] Initial dosages can also be formulated by comparing the
effectiveness of the compounds used in the methods of the present
invention in model assays with the effectiveness of known
compounds. For instance, initial dosages can be formulated by
comparing the effectiveness of the compounds in model assays with
the effectiveness of other compounds that have shown efficacy in
treating the present conditions. In this method, an initial dosage
can be obtained by multiplying the ratio of effective
concentrations obtained in the model assay for the compounds used
in methods of the present invention and the control compound by the
effective dosage of the control compound. For example, if a
compound useful in a present method is twice as effective in a
model assay as a known compound (i.e., the EC50 of the compound is
equal to one-half the EC50 of the known compound in the same
assay), an initial effective dosage of the compound would be
one-half the known dosage for the known compound. Using these
initial guidelines one having ordinary skill in the art could
readily determine an effective dosage in humans or other
mammals.
[0106] Dosage amount and interval may be adjusted individually to
provide levels of the active compound which are sufficient to
maintain therapeutic effect. One having skill in the art will be
able to optimize therapeutically effective local dosages without
undue experimentation.
[0107] 4. Combination with Other Anti-Inflammatory Agents
[0108] In pharmaceutical dosage forms, the present methods can use
formulations where compounds are administered in the form of their
pharmaceutically acceptable salts, or they may also be used alone
or in appropriate association, as well as in combination with other
pharmaceutically active compounds. Agents of particular use in the
formulations of the present invention include, for example, local
anesthetics, counterirritants, anti-inflammatory agents, or any
agent that has a therapeutic effect for inflammatory diseases or
conditions.
[0109] The preferred anti-inflammatory agents include, but are not
limited to, prescription and nonprescription topical and aerosol
corticosteroids, non-steroidal anti-inflammatory agents including
salicylates, colchicine, para-aminophenols, propionic acids,
macrolide immunosuppressives, dapsone, clobetasol, halobetasol,
diflorasone, piroxicam, ketorolac, ketoprofen, indomethacin and
specific cyclooxygenase inhibitors.
[0110] The preferred counterirritants include, but are not limited
to, glycerol, corticosteroids and salicylates. The preferred
anesthetics include, but are not limited to, amide caines and
counterirritants with lidocaine, cocaine, bupivicaine, mepivicaine,
etidocaine, chloroprocaine, proparacaine, tetracaine, benzacaine,
prilocaine, benoxinate, dibucaine, dyclonine, pramoxine, menthol,
resorcinol, thymol and camphor.
[0111] Any other compound that has potential efficacy in the
treatment of the present conditions can also be used.
[0112] 5. Routes of Administration
[0113] The compositions useful in the present methods can be
administered to a patient using a variety of routes, such as oral,
parenteral or local routes. The present compositions are typically
administered to a patient as a local application, where "local
application," or "locally applied," refers to the administration of
a composition at the local site of the disease, whether by local
injection, topical administration, or any such method that results
in a relatively high concentration of the compounds used in methods
of the present invention at the site of the disease. As such,
administration of the compounds can be achieved in various ways,
including by topical application of the composition to the site of
the disease or condition, i.e., direct application of a formulation
to the affected skin or mucous membrane. In addition, compositions
can be formulated for injection and injected locally at the site of
the disease or condition, e.g., local subcutaneous injection at the
site of the disease.
[0114] 6. Areas for Topical Application
[0115] The compositions usefil in the present methods can be
applied to any site of any of the present conditions, including
localized conditions or conditions affecting large areas of the
body or even covering the entire body, can be applied to the skin
and/or to mucous membranes, and can be applied to any affected part
of the body, including the face, forehead, chin, eyes, eyelids,
eyebrows, nose, skin near the nose, cheeks, ears, mouth, tongue,
inside of the cheeks, gums, head, hair, scalp, neck, chest, back,
lower back, armpit, skin folds of armpit, elbow, elbow fold,
wrists, ankles, legs, arms, insides of wrists, insides of arms,
nails, knees, area behind knees, hands, feet, palms, soles,
fingers, toes, genitals, or any other affected part of the
body.
[0116] 7. Frequency of Administration
[0117] The compositions useful in the present methods can be
administered one time or multiple times, depending on the compound,
the severity of the condition, and the initial response of the
condition to the treatment. For example, the compositions can be
administered 1, 2, 4, or more times per day, and can be
administered every 1, 2, 4, 7, or more days. Such treatments can be
administered for a limited duration, or indefinitely until the
condition has resolved. The compositions can be applied locally as
a "leave on" product, meaning that the composition is applied to
the patient and allowed to remain indefinitely at the site of
application, or as a "wash off" product, meaning that the
composition is allowed to remain at the site of application for a
limited amount of time, e.g., for a certain number of seconds,
minutes, hours, etc.
[0118] It will be appreciated that the present methods of treatment
can be applied alone or in combination with other surgical or
non-surgical treatments for these conditions.
IV. THERAPEUTIC APPLICATIONS
[0119] As discussed previously, the formulations and methods of the
invention are useful for treating inflammatory diseases or
reactions. Preferably, these inflammatory diseases are disorders of
the bone joint. In especially preferred embodiments, these methods
are used to treat inflammatory diseases such as osteoarthritis,
osteoporosis, rheumatoid arthritis, and soft tissue rheumatism. The
term "treatment" is used to refer to a reduction of any of the
symptoms associated with these inflammatory conditions, including,
but not limited to, inflammation, redness, pain, swelling,
lameness, and loss of mobility.
[0120] The methods of the present invention can also be used to
prevent and/or treat inflammatory skin diseases (e.g., atopic
dermatitis, eczema, contact dermatitis, allergic dermatitis), skin
irritation, inflammatory pulmonary disease or reactions (e.g.,
asthma, allergic rhinitis, chronic obstructive pulmonary disease,
and adult respiratory distress syndrome), inflammatory
musculoskeletal disease or reaction (e.g., soft tissue rheumatism,
exercise-induced injury, rheumatoid arthritis, psoriatic arthritis,
osteoporosis and osteoarthritis), inflammatory gastrointestinal
disease or urogenital reaction (e.g. enterocolitis, gastritis,
Crohn's disease, interstitial cystitis, vaginitis, and ulcerative
colitis), autoimmune disease or reactions (e.g., inflammatory bowel
disease, and psoriasis), and transplantation rejection
reactions.
[0121] Furthermore, the compounds of the invention can be used to
treat lameness or loss of mobility arising from the inflammatory
musculoskeletal diseases described above, as well as other
conditions. In another preferred embodiment, the methods of this
invention are used to treat pain in or around a bone joint arising
from the inflammatory musculoskeletal diseases described above, as
well as other conditions.
EXAMPLES
[0122] The following examples are offered to illustrate, but not to
limit the claimed invention.
Example 1
[0123] This example illustrates the effectiveness of fortified
formulations of stabilized rice bran derivatives for reducing
lameness and joint inflammation in horses.
[0124] Clinical trials on 51 horses were conducted to determine the
ability of formulations of this invention to improve lameness and
reduce joint inflammation. Horses were fed with formulations of
this invention and other commercially available treatments used to
treat lameness. Horses were videotaped on day 1, day 21, and day 50
of the trial. Lameness was evaluated using the scoring system
illustrated in FIG. 1. The correlation between treatment with the
various formulations and "lameness" is illustrated in FIG. 2. The
effect of treatment with the various formulations on inflammation
was measured by injecting the neck of the horse with carrageenan on
day 21 and day 50, then observing the size of the injection site
over 7 hours (FIG. 3). Prior to initiation of the trial, a range-
finding study was conducted to determine the amount of carageenan
and route of administration necessary to induce measurable amounts
of inflammation.
[0125] A range finding study was conducted to find quantifiable
endpoint of carrageenan induced subcutaneous skin inflammation and
to compare various commercially available oral products designed to
reduce inflammation in horses and to then compare these to
phenylbutazone administration intravenously 12 hours prior to
comparison and again one hour to commencement of comparison, on day
21 and day 50 post commencement feeding of the oral preparations. A
0.1 mL of subcutaneous injection of 2% carrageenan in sterile
water, autoclaved for sterility provided a reproducible challenge
under the study conditions utilized. A dose of 2 grams of
phenylbutazone given intravenously twice prior to each evaluation
day provided by a clinically significant reduction in inflammation
for comparison.
[0126] Preparation of potential horses for the project consisted of
evaluation of general health and history of clinical health and
soundness. Where pertinent, a routine lameness evaluation to verify
presence of arthritic and/or lameness of long-standing duration
rather that short-term lameness (bruises, soft tissue damage, etc.)
that would heal with or without intervention. If an individual
satisfied these criteria, then routine immunizations (tetanus
toxoid, eastern-western encephalitis influenza, and
rhino-pneumonitis) as well as cost to the owners. Once the trial
cell was completed, containing three of five clinically lame horses
(exception trials cells "A" and "B"), then each individual was
given a project number for identification. Each number was then
branded into the antero-lateral aspect of the hoof was on the left
front hoof for identify verification on days #1, day#21 and day
#50.
[0127] Fifty one horses (10 groups of 5 each with one extra) were
fed a basal diet consisting of alfalfa and/or grass hay, oats and
stabilized rice bran as needed for consumption of the various oral
products being evaluated. Horses were not randomly selected for the
trial cells, since at least three of five in each cell had to
exhibit clinical lameness. (Cells "A" and "B" excluded from this
requirement) Horses once identified as a part of the trial were
eliminated if (1) they refused to eat the product (2) were given
medication other than those included in the project, for whatever
reason that might interfere with the results.
[0128] The purpose of this study was to compare the various
products, both available and non-available, alone or in combination
with each other and against phenylbutazone, the "benchmark" NSAID
for tissue inflammation reductions. In addition, videos of lame
horses were taken on day #1, #21, and #50 for subjective
evaluation. Looking for improvement in the lameness condition
presented on day #1.
[0129] A. Materials and Methods
[0130] A range finding study was conducted to find quantifiable
endpoint of carrageenan-induced subcutaneous skin inflammation and
to compare various nutraceutical products both commercially
available and yet-to-be available to phenylbutazone, the
"benchmark" anti-inflammatory drug in equine veterinary
medicine.
[0131] In determining a quantifiable endpoint for carrageean
injection 200 mL of a 2% w/v sterile preparation was made by
certified compounding pharmacist, using autoclave method for final
sterilization. Six horses were chosen for initial evaluation of the
product. An area approximately 8 cm.times.24 cm was clipped on the
side of the neck mid-cervical and approx. 3 cm below the mane and
upon whichever side the mane flows. The area was scrubbed with 70%
alcohol-essential oil skin preparation. Immediately six small dots
were placed using waterproof marker in the clipped area
approximately equidistant horizontally along a line tuberculosis
syringe using a 25 gauge 5/8" needle inserted through the rubber
stopper previously saturated with 70% alcohol and allowed to
partially dry. In addition, 0.2 mL. increment of 2% carrageean
(w/v) was drawn into 5 subsequent tuberculosis syringes and hub
could thus be accounted. Animal was then retrained and a 0.1 cc
increment was then placed intradermally under each skin mark,
starting the 25 gauge 5/8" needle approximately {fraction (3/16)}"
from the mark in a scrubbed area and ending with the point of the
needle under the indelible mark. No surgical gloves were used, but
care was taken to avoid post-scrub skin contamination. All six
horses were injected intradermally as described for dogs in the
canine DCV project. Four of the six horses exhibited inflammatory
lumps of 12.times.16 mm to 18.times.22 mm in size which enlarged in
an elliptical fashion initially within 2-3 hours, but did not
regress measurably past the initial size even after several weeks.
The lumps seemed to be mildly to moderately tender with the horses
apprehensive when even slight digital pressure applied to the test
area. The initial swelling induced by the sterile water
approximated the carrageean swellings, but regressed to near and
normal within several hours. Since the swellings induced did not
fit the trial protocol requirement s, a second route of
administration was performed on 6 different horses. In the second
group of range findings study horses, only route of administration
changed. All 0.1 cc injections were placed subcutaneously rather
intradermally. Restraint again was important as any carrageenan
laced along the route of administration upon needle withdrawal
tended to induce a lingering bleb as well as initial measurable
swelling post injection at the sire of needle induction. In some
instances this was unavoidable under best of circumstances and
restraint due to normal equine response to needle penetration and
material placement under the cutaneous later of skin. In other
instances, some micro bleeding at injection site was unavoidable
and apparent. In each case, swelling was measured and recorded,
since micro bleeding tends to resolve within several hours and
certainly in less than 5 hours in each instance.
[0132] Swelling resulting from subcutaneous placement of 2%
carrageenan were mostly elliptical, measurable, and transient.
Sites ranged from 0 mm.times.Omm, initially to 45 mm.times.60 mm
after 7 hours.
[0133] After 14 days post range finding evaluation, three of the
six horses initially used were then chosen at random, administered
intravenous phenylbutazone at 1.0 gms. Per 500 pounds body weight
and again challenged with 0.1 cc of 2% carrageenan in five sites,
using 0.1 cc sterile water as a control. All injections were given
subcutaneously as previously described. Size of resulting lumps was
measured immediately and again at hourly intervals for 7 hours.
Resulting swellings were minimal, measurable, and provided a
comparison to phenylbutazone given at the recommended dose and
frequency generally used. Note however that phenylbutzone cannot be
used on a race day at racetracks sanctioned for public wagering. In
addition AHSA (American Horse Association) sanctioned shows have
specific guidelines which specify when and how much phenylbutazone
may be administered to a competing horse. This initial evaluation
as well as follow-up evaluation during the test trials suggests the
benefits of phenylbutazone as it may be used to satisfy
drug-testing policy may be long gone prior to competition. This
makes the benefits derived from oral nutraceuticals all the more
important and meaningful in controlling inflammation. The
transitory nature of phenylbutazone benefit when used under present
guidelines places it more in the "band aid therapy" category than
previously recognized by veterinarians and horsemen alike.
Nevertheless it will remain as the standard by which most responses
to inflammation/lameness treatments are measured likely for years
to come.
[0134] Fifty-one horses were selected for ten treatment groups.
Horses picked were selected at random for age, use, breeding,
weight and sex. Age ranged from 1 year to 37 years of age. Use
included retired, broodmares, herd sires, western performance,
hunter-jumper performance, in early training, and untrained.
Breeding included quarter horse and quarter horse cross,
thoroughbred cross, appaloosa, and paints. Weight varied from
approximately 700 lbs to 1200 lbs. There were 7 intact males, 27
castrated males, and 17 intact females. Horses were housed under
typical upper-scale horse facilities in two climates with 26 head
in northwestern Montana and 25 head in central Arizona, USA.
Ambient temperatures during the trial period ranged from 5 F to 106
F. All the products were stored at 73 F to 82 F and fed at ambient
temperatures. All attempt was made to utilize horses typical of
those eventually targeted for the product being evaluated, and
under conditions typically seen when products comes to market.
5 Equiflex Glucosamine Phenylbut. Carageen. Treatment Group Animals
PL-100 fed. A. b. admin. c. admin. A 5 NO NO NO NO YES B 5 NO NO NO
YES YES C 5 NO NO COSEQUIN NO YES D 5 NO NO NEXT LEVEL NO YES E 5
15 gm/day YES YES NO YES F 5 15 gm/day YES YES NO YES G 5 30 gm/day
YES YES NO YES H 5 30 gm/day NO YES NO YES I 5 NO YES YES NO YES J
5 NO YES YES NO YES liquid only A. loading dose will be double dose
for 21 days maintenance dose will be single dose for 30 days b.
Cosequin and next level given at maximum recommended dose c.
Phenylbutazone given at 1 gm/500 lbs 12 hours prior to challenge
and 1 gm/500 lbs 1 hour prior to challenge with carrageean Cells to
contain minimum of 3 clinically lame horses with the exception of
cells "A" and "B".
[0135] Fifty-one horses were selected for ten treatment groups.
Horses picked were selected at random for age, use, breeding,
weight and sex. Age ranged from 1 year to 37 years of age. Use
included retired, broodmares, herd sires, western performance,
hunter-jumper performance, in early training, and untrained.
Breeding included quarter horse and quarter horse cross,
thoroughbred cross, appaloosa, and paints. Weight varied from
approximately 700 lbs to 1200 lbs. There were 7 intact males, 27
castrated males, and 17 intact females. Horses were housed under
typical upper-scale horse facilities in two climates with 26 head
in northwestern Montana and 25 head in central Arizona, USA.
[0136] B. Purpose of the Study
[0137] 1.0 A Study Conducted by White Eagle Toxicology Laboratories
in Doylestown, Pa.
[0138] Under direction of DVC Biologics, L.P. of Wilmington, Del.
described the effect of egg powder on acute carrageean-induced
inflammation in female beagle dogs. Findings indicated a
significant reduction in inflammation. The purpose of this study
was to compare reduction in inflammation induced by carrageean
resulting from various commercial nutraceuticals claiming
inflammation reduction with egg powder and egg powder and Equiflex
combinations using Phenylbutazone as a benchmark comparison
anti-inflammatory in the horse.
[0139] 2.0 Study Schedule
[0140] Study initial date--July, 2000 Experimental
termination--November, 2000
[0141] 3.0 Test System:
[0142] 3.1 Species: Horse
[0143] Breed: Qrt, Qrtx, Qrt-tbx, Appaloosa, Paint
[0144] 3.2 Supplier: Randy Johnson Qrt Horses Diamond B Ranch Mark
ArnoldGary D. Kaufman, D. V. M Scott Davis
[0145] 3.3 Age: 1 year to 37 year
[0146] 3.4 Number:51
[0147] 3.6 Justification for test system selection: Horses in use
in typical life-style of those anticipated as target group for
commercial product.
[0148] 3.7 Justification for the number of animals: This study was
designed to use the fewest number of horses possible consistent
with the objective of the study and scientific needs.
[0149] 3.8 Justification for route of administration: The test
product is intended to be administered as a supplement to the basal
diet.
[0150] 3.9 Acclimation: All animals were acclimated for at least 2
weeks prior to the study.
[0151] 3.10 Selection Criteria
[0152] All the horses were evaluated for general health and
lameness if present. Horses were selected if they healthy, not
likely to require medication for the next two months, and at least
exhibit lameness from chronic rather than acute inflammatory
changes.
[0153] 4.0 Animal Husbandry:
[0154] 4.1 Housing: Horses were housed in individual stalls, runs,
pastures, or combination three.
[0155] 4.2 Environmental conditions: The horses maintained
according to currently acceptable practices of good animal
husbandry.
[0156] 4.3 Food: Alfalfa and/or grass hay with/without oats and
supplemented with a stabilized rice bran as needed to consume test
product.
[0157] 4.4 Water: Available ad libitum
[0158] 4.5 Non contaminants were known to be in the feed or water
that would be expected to alter to outcome of the study.
[0159] 5. Animal Identification and Randomization:
[0160] Each horse was individually identified using a hoof brand
number burned into the anterolateral aspect of the left front hoof.
Horses for each treatment group were not randomly assigned since
three of 5 in groups "C" through "J" required a clinical
lameness
[0161] 6.0 Test products:
[0162] 6.1 Identity: Egg powder, "Cosequin", "Next Level", and
Equiflex.
[0163] 6.2 Source of test product: Wolcott Farms, RiceX, DVC
Biologics, Nutramax Laboratories & Sure Nutrition.
[0164] 6.3 Storage conditions/stability: Test products were stored
at room temperatures in a dry place protected from sunlight.
Stability of the products under conditions of storage as
stated.
[0165] 6.4 Disposition of the test products: At the conclusion of
this study all unused portions are being held or disposed of as
directed by study director.
[0166] 7.0 Study Design
[0167] 7.1 Range finding study: A range finding study was conducted
to: (1) establish a measurable carrageenan induced inflammation on
the neck under the mane and (2) to verify that phenylbutazone given
in an accepted manner would significantly reduce inflammation
caused by carrageenan to use as a comparison.
[0168] During attempts to establish a measurable carrageenan
induced inflammation using route of administration suggested by DCV
Biologics. Multiple attempts were unsuccessful. Inflammation did
not show a patterned increase in size of bleb and transitory
swelling suggested. Rather it result in a small painful lump that
frequently did not increase nor dissipate in any set pattern, but
rather lingered for weeks and sometimes months as a painful,
obvious swelling.
[0169] A second model was evaluated using subcutaneous route of
administration with all other parameters being identical. A
transitory lump was produced with almost all evidence being gone in
24-40 hours. In addition, the swellings were quite measurable over
7 hour period, increasing and in some instances starting to
decrease within that time frame.
[0170] The swellings were for the most part elliptical or at least
not perfectly round. Inflammation in horse skin id often manifest
in a rash" appearance rather than round lumps, so the shape of
swelling was of no real surprise. Measurements were taken using
basically height x width of the swellings to establish a more
representative number to describe the reaction to the
carrageenan-induced inflammation. It should be noted that swelling
is a 3 dimensional phenomenon as each lump had thickness as well as
height and width. In some horses, measurements went from a
significant width x height to 0.times.0. In these cases the
swelling melted away first in the middle of the lump and eventually
dissipated at the borders to the extent no readings were possible.
This again is not surprising, since skin swellings in horses often
vanish in this manner. Strictly reading the numbers in these cases
may be quite confusing. In this trial, swellings were simply
identified, measured and numbered and when identification was no
longer possible, measure was called 0.times.0.
[0171] 7.2 Efficacy study of the tests products: Al tests products
were measured using and electronic gram scale for solids and
graduated dose syringe for liquids. Each product was measured and a
daily amount was measured was placed in an individual plastic
container and heat-sealed using a Food saver Bag Vac Sealer. All
liquids were placed in a bag within a bag and heat-sealed. Each bag
was then labeled with the horse's name and identification number.
Each product was mixed as requested under protocol treatment chart.
Only the product to be given from day 1 to day 21 was issued to
complete the trial. Treatment group "J" did not receive daily
liquid Equiflex after day 21 challenge simply because product did
not arrive form Rice-X as requested in a timely fashion. Upon
arrival, product was fed as a loading dose for 6 days to compensate
and ultimately allow for total amount of product in the horses by
day 51. This variance was discussed with and approved by Win of
Wolcott Farms as the logical correction to make. Cosequin was fed
twice daily three level scoops each time. This was continued
throughout the entire 51 days rather than drop to maintenance dose,
since the label suggests that the horse should be watched for
"movement and attitude" and that maintenance or transitional dosage
of a lesser amount "may be increased at any time if needed". Rather
than get it an issue, since comparisons get into an issue, since
comparisons were being drawn, Cosequin was fed throughout at
maximum recommended dose. Next level was fed at maximum dose of 2
oz throughout the first 21 days rather than the first 10 days, and
then dropping to 1 oz per day as the label suggests. Again this was
to ensure maximum results. Egg biologics was fed at 15 gm/day and
30 gm/day. Each was doubled during the first 21 days. When used in
combination with equiflex/glucosamine both egg biologics and
equiflex/glucosamine were doubled in amount. Equiflex was fed at a
dose of 2.75 oz as a standard dose and a 2.72 oz when fed with egg
biologics. Liquid Equiflex was fed 4.25 oz/day maintenance, 8.5
oz/day loading dose.
[0172] 8.0 Antemortem Observations
[0173] 8.1 Local observations: Each animal was observed at least
twice a day during feeding time and any changes noted. #59 Bowie
was eliminated from the trials after day #5 for refusal to eat the
loading dose of 60 gms/day egg biologics with 5.44 oz day Equiflex
powder. #7 Dolly was given intravenous flunixamine on day 20 and
scratched from the day 21 evaluation, but included in the day 50
evaluation. #1 Copper was eliminated from the trial after
carrageenan included lumps persisted on his neck for more than two
weeks post-injection. #18 Mercedes was eliminated on day 23. He
required continuous medication for gastric ulcers, which could
potentially alter trial results.
[0174] 8.2 Data collection: All measurements were taken by the
study director and recorded by support personnel. Measurements were
made hourly for 7 hours with most being on the hour with few
varying several minutes, depending on the difficulty experienced in
restraining fractious/dangerous animals.
6 TITLE: Evaluation Of The Anti-Inflammatory Efficacy Of PL-100 (a
whole egg product, ex DCV), `Arthreflex`, `Next Level` and
`Cosequin` on Horses versus `Bute` PURPOSE: Assess the statistical
and clinical efficacy of two test products (PL-100 in an inert
carrier base and in combination with `Arthreflex`) versus three
market controls in i) an equine model of inflammation, and ii)
clinically lame horses. ANIMALS: SPECIES: Equine STRAIN/BREED:
Variety - To be documented SEX: To be documented AGE: 3 mo-25 years
SOURCE: to be documented Method of IDENTIFICATION: Total NUMBER: 50
Treatments: 10 Number per 5 horses per TEST GROUP: treatment Number
of 1 REPLICATES : Type and Stabled horses description of HOUSING:
Husbandry, All horses are vaccinations, immunized general
management: according to local standard prior to starting RATION/
BASAL DIETS DIET: Source: Alfalfa roughage diet with additional
grain Nutrient Protein 14%- Specifications: 20% Basal diets are
based upon NRC Formula: Nutritionally adequate and on that is
commonly utilized for feeding the type of horse used in the trial.
Medication: None EXPERIMENTAL Total # 50 Horses: DESIGN: Total #
random Males/Females: Total # 10 Treatment Groups: Total Duration
of 50 days Treatment: Method of To be documented Randomization:
Blinding: None Statistical Student's t-test, Analysis: ANOVA &
Bonferroni's Multiple Comparison post-test.
[0175]
7 TREATMENT GROUPS: Treatment #'s PL-100 Phenylbutazone Carrageenan
Group Animals Fed.sup.a Arthreflex Glucosamine.sup.b
Administered.sup.c Administered A 5 No No No No Yes B 5 No No No
Yes Yes C* 5 No No Cosequin No Yes D* 5 No No Next Level No Yes E*
5 15 gm/day Yes Yes No Yes F* 5 15 gm/day No Yes No Yes G* 5 30
gm/day Yes Yes No Yes H* 5 30 gm/day No Yes No Yes I* 5 No Yes Yes
No Yes J* 5 No Equiflex Yes No Yes Liq .sup.aLoading dose will be
bid for 21 days Maintenance dose will be sid for 30 days .sup.b1800
mg twice a day .sup.cto be documented *Cells to contain minimum of
3 Clinically Lame Horses
[0176] Experimental Design:
[0177] Rangefinding Study: A rangefinding study was conducted to
establish a quantifiable end point of carrageenan induced
inflammation and to determine the dose of phenylbutazone which will
reduce the inflammation by approximately 50%. Based on these
results, the procedure for challenge will be as follows:
[0178] An area on the body will be clipped and six small lines will
be drawn on the neck with marker. Five (5) 0.1 mL intradermal
injections of 2% carrageenan in sterile water will be made in the
clipped area and a single injection of water will serve as a
control. The diameter of the injection sites will be measured with
calipers and recorded immediately after injection and hourly there
after for 7 hours following injection. 5 test sites and 1 control
(water) per horse, therefore 25 test sites per test cell+5
controls
[0179] All PL-100/Glucosamine treatments will be mixed by DuCoa
prior to application and shipped to the test facility. Retention
samples of PL-100 will be retained for assay by DCV. Retention
samples for all products will be retained for assay by WFY. Prior
to initialization of the trial, horses will be selected the basis
of comparable size and general good health. Animals will be grouped
by treatment as described in the treatment table above. Horses
given PL-100 will receive a loading dose (defined as the
maintenance dose given twice a day) for 21 days. Horses on market
control products will follow manufacturer's instructions. On day 21
all horses will be challenged with carrageenan and evaluated for
inflammatory response. Ideally, the same person should do the
intradermal injections and the same person should do the measuring
of the inflammatory response. Starting on day 22, the horses
receiving PL-100 will be given PL-100 once a day as a maintenance
dose. All horses will then be challenged with carrageenan on day
50
[0180] Clinically Lame Horses--Observational Methods
[0181] Prior to the start of the trial each clinically lame horse
will be exercised recorded by videotape. The video evidence should
clearly illustrate the animals' stride, extension and general
mobility. The same record of the animals' physical exhibit should
also be kept at day 21 and day 50 of the trial. The performance
difference should be evaluated across these three intervals by the
same evaluator. The evaluators' comments and observations will be
part of the trial record. The same evaluator should be used for
each of the clinically lame horses.
8 SCHEDULE: Date Treatments August (day 0) House horses Start trial
feeding Size test August (day 21) Challenge horses September (day
50) Measure results
[0182]
9 DATA MEASUREMENTS & RECORDS: Data Collected or Measurement
Record daily observations for general health and disease status
throughout trial period Record reasons for any removal of animals
& specific exclusion criteria met Record disposition of all
animals and materials and attach receipts
[0183] METHOD OF Group mean values (+Std. Devns.). Pre- vs
Post-comparisons within DATA ANALYSIS each treatment group conduct
using paired Student's t-test. Between & STATISTICS: group
comparisons using one-way ANOVA. Where significant differences
(p.ltoreq.0.05) are seen, use Bonferroni's Multiple Comparison
post-test to determine which means are significantly different from
each other.
Example 2
[0184] This example sets forth preferred formulations of this
invention.
10 Human Formulations NutraFlex .TM. (For Humans) Serving Dose 10
grams/day RiSoluble .TM. 79% .+-. 5 Yucca 10% .+-. 1 Glucosamine
Sulfate 5% .+-. 1 N-Acetylglucosamine .sup. 3% .+-. 0.3
Methylsulfonylmethane (MSM) .sup. 2% .+-. 0.3 Grape seed extract
.sup. 1% .+-. 0.3 NutraFlex: Drink NAG .sup. 3% .+-. 0.5 MSM .sup.
2% .+-. 0.3 Yucca 10% .+-. 1 Glucosamine Sulphate 5% .+-. 1 Grape
seed ext .sup. 1% .+-. 0.1 Boswellin .sup. 1% .+-. 0.1 Solubles 78%
.+-. 5 Dose 10.0 g/day NutraFlex: Capsules RiSolubles 28.0% .+-. 3
N-Acetyl Glucosamine 11.11% .+-. 1 Glucosamine sulphate 16.67% .+-.
2 Methyl Sulfonyl Methane 11.11% .+-. 2 Grape seed extract 5.58%
.+-. 1 Yucca Concentrate (70%) 5.58% .+-. 1 Hemp seed oil 5.58%
.+-. 1 Boswellin 5.58% .+-. 1 Ginger powder 5.58% .+-. 1 Curcumin
(95%) 5.58% .+-. 1 Total 1.80 g for 4 capsules two with breakfast
and two with dinner NutraFlex: Bar Solubles 48.39% .+-. 3 N-Acetyl
Glucosamine 8.06% .+-. 2 Glucosamine sulphate 8.06% .+-. 2
Methylsulfonylmethane 16.13% .+-. 2 Yucca concentrate 3.23% .+-.
0.5 Grape seed extract 3.23% .+-. 0.5 Aswagandha 8.06% .+-. 0.7
Ginger powder 3.23% .+-. 0.5 Curcumin 1.61% .+-. 0.3
[0185]
11 Formulations for Horses EquiFlex .TM. (For Horses) Serving size
2.75 oz/day (78 grams/day) Stabilized Rice Bran 88.47% .+-. 5 Yucca
5.13% .+-. 1 Glucosamine Sulfate 3.84% .+-. 0.5 N-Acetylglucosamine
1.28% .+-. 0.3 Methylsulfonylmethane (MSM) 1.28% .+-. 0.3 Total
100% EquiFlex-Liquid Formulation Rice Bran Oil 30-60% Glucosamine
Sulphate 2.62% .+-. 0.5 N-Acetylglucosamine 0.87% .+-. 0.2
Methylsulfonylmethane 0.87% .+-. 0.2 Yucca Concentrate (70%) 3.5%
.+-. 0.3 Water 30-60% Gum (Emulsifier) 0.2% .+-. 0.05 Sodium
benzoate(Stabilizer) 0.15% .+-. 0.03 Citric acid (Preservative)
0.6% .+-. 0.1 Apple flavor 0.25% .+-. 0.05 Dose 155 g/day
[0186] Topical Formulations
[0187] Arthritis Cream
[0188] Distilled Water
[0189] EPA
[0190] Toco
[0191] Emu Oil
[0192] Stabilized Oxygen
[0193] Orange Oil
[0194] Silica
[0195] Colloidal Silver
[0196] Lemon Oil
[0197] Poly Base
[0198] Yucca
[0199] Glucosamine sulfate
[0200] Methylsulfonylmethane
[0201] All publications, patents and patent applications mentioned
in this specification are herein incorporated by reference into the
specification in their entirety for all purposes. It is understood
that the examples and embodiments described herein are for
illustrative purposes only and that various modifications or
changes in light thereof will be suggested to persons skilled in
the art and are to be included within the spirit and purview of
this application and scope of the appended claims. All
publications, patents, and patent applications cited herein are
hereby incorporated by reference in their entirety for all
purposes.
* * * * *