U.S. patent application number 17/666305 was filed with the patent office on 2022-09-22 for compositions and methods for acutely raising nitic oxide levels.
This patent application is currently assigned to Nature's Sunshine Products, Inc.. The applicant listed for this patent is Nature's Sunshine Products, Inc.. Invention is credited to John G. Babish, Clinton J. Dahlberg, Wei Gao, Mohan Kaadige, Matthew L. Tripp.
Application Number | 20220296552 17/666305 |
Document ID | / |
Family ID | 1000006391077 |
Filed Date | 2022-09-22 |
United States Patent
Application |
20220296552 |
Kind Code |
A1 |
Tripp; Matthew L. ; et
al. |
September 22, 2022 |
COMPOSITIONS AND METHODS FOR ACUTELY RAISING NITIC OXIDE LEVELS
Abstract
Disclosed herein are compositions and methods for acutely
raising nitric oxide levels in a subject. In one example, the
composition can include, an effective amount of a NOS dependent
source of nitric oxide; an effective amount of a NOS independent
source of nitric oxide; and an effective amount of a
myeloperoxidase inhibitor; wherein the composition acutely raises
nitric oxide levels in a subject above a level provided by the
available sources of nitric oxide in the subject prior to
administration of the composition. Further presented is a method of
treating a subject for a condition or disorder that is response to
nitric oxide therapy, including: acutely raising nitric oxide
levels in a subject by simultaneously increasing biosynthesis of
nitric oxide, increasing nitrate/nitrite levels, and inhibiting
meyloperoxidase activity.
Inventors: |
Tripp; Matthew L.; (Camano
Island, WA) ; Dahlberg; Clinton J.; (Saratoga
Springs, UT) ; Babish; John G.; (Brooktondale,
NY) ; Kaadige; Mohan; (Salt Lake City, UT) ;
Gao; Wei; (Lehi, UT) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Nature's Sunshine Products, Inc. |
Lehi |
UT |
US |
|
|
Assignee: |
Nature's Sunshine Products,
Inc.
Lehi
UT
|
Family ID: |
1000006391077 |
Appl. No.: |
17/666305 |
Filed: |
February 7, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15756548 |
Feb 28, 2018 |
11241406 |
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PCT/US2016/049306 |
Aug 28, 2016 |
|
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17666305 |
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62211673 |
Aug 28, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 36/82 20130101;
A61K 31/51 20130101; A61K 36/63 20130101; A61K 36/185 20130101;
A61K 36/87 20130101; A61K 31/555 20130101; A61K 36/534 20130101;
A61K 36/61 20130101; A61K 31/198 20130101; A61K 36/42 20130101 |
International
Class: |
A61K 31/198 20060101
A61K031/198; A61K 36/87 20060101 A61K036/87; A61K 36/82 20060101
A61K036/82; A61K 36/63 20060101 A61K036/63; A61K 36/185 20060101
A61K036/185; A61K 36/42 20060101 A61K036/42; A61K 31/51 20060101
A61K031/51; A61K 31/555 20060101 A61K031/555; A61K 36/534 20060101
A61K036/534; A61K 36/61 20060101 A61K036/61 |
Claims
1. A method of treating a subject for a condition or disorder that
is responsive to nitric oxide therapy, comprising: acutely raising
nitric oxide levels in a subject by simultaneously increasing
biosynthesis of nitric oxide, increasing nitrate/nitrite levels,
and inhibiting meyloperoxidase activity.
2. The method of claim 1, wherein the condition or disorder is a
nitric oxide related pathology.
3. The method of claim 2, wherein the nitric oxide related
pathology comprises a member selected from the group consisting of
Alzheimer's disease, angina, asthma, congestive disorders, Crohn's
disease, deep vein thrombosis, dementia, diabetes (types, 1, 2 and
3), diabetic foot disorders, diminished exercise capacity,
endothelial dysfunction, endotoxemia, erectile dysfunction,
fibromyalgia, heart attack, heart failure, hypertension,
inflammatory bowel disease, leaky gut, macular degeneration,
monocyte-mediated arterial plaque formation, motor dysfunction,
multiple sclerosis, obesity, oxidation of LDL, peridontal disease,
peripheral arterial disease, platelet stickiness, portal
hypertension, pregnancy/pre-eclampsia, premature ejaculation,
pulmonary hypertension, Raynaud's disease, renal failure, sleep
apnea, smooth muscle cell proliferation, stroke, and
vasculitis.
4. The method of claim 1, wherein the condition or disorder is a
cardio-metabolic disorder.
5. The method of claim 4, wherein the cardio-metabolic disorder
comprises a member selected from the group consisting of:
Alzheimer's disease, angina, asthma, congestive disorders, Crohn's
disease, deep vein thrombosis, dementia, diabetes (types, 1, 2 and
3), diabetic foot disorders, diminished exercise capacity,
endothelial dysfunction, endotoxemia, erectile dysfunction,
fibromyalgia, heart attack, heart failure, hypertension,
inflammatory bowel disease, leaky gut, macular degeneration,
monocyte-mediated arterial plaque formation, motor dysfunction,
multiple sclerosis, obesity, oxidation of LDL, peridontal disease,
peripheral arterial disease, platelet stickiness, portal
hypertension, pregnancy/pre-eclampsia, premature ejaculation,
pulmonary hypertension, Raynaud's disease, renal failure, sleep
apnea, smooth muscle cell proliferation, stroke, and
vasculitis.
6. The method of claim 4, wherein the cardio-metabolic disorder
comprises a member selected from the group consisting of
hypertension, cardiovascular dysfunction, neurodegeneration,
arthritis, asthma, and septic shock.
7. The method of claim 4, wherein the cardio-metabolic disorder
comprises preventing the formation of arterial plaque.
8. The method of claim 4, wherein the treating of the subject is
prophylactic.
9. The method of claim 1, wherein the condition or disorder is a
myeloperoxidase related pathology.
10. The method of claim 9, wherein the myeloperoxidase-related
pathology is Alzheimer's disease, angina, asthma, general
congestive disorders, Crohn's disease, deep vein thrombosis,
dementia, diabetes (types, 1, 2 and 3), diabetic foot disorders,
diminished exercise capacity, endothelial dysfunction, endotoxemia,
erectile dysfunction, fibromyalgia glomerulonephritis, heart
attack, heart failure, hypertension, immune deficiency,
inflammatory bowel disease, leaky gut, macular degeneration,
monocyte-mediated arterial plaque formation, motor dysfunction,
multiple sclerosis, obesity, oxidation of LDL, peridontal disease,
peripheral arterial disease, platelet stickiness, portal
hypertension, pregnancy/pre-eclampsia, premature ejaculation,
pulmonary hypertension, Raynaud's disease, renal failure, sleep
apnea, smooth muscle cell proliferation, stroke, vasculitis and
diseases associated with skin such as slow wound healing, wrinkles,
and premature signs of aging.
11. The method of claim 9, wherein the myeloperoxidase related
pathology comprises increased oxidized LDL cholesterol.
12. The method of claim 9, wherein the myeloperoxidase related
pathology comprises metabolic syndrome, type 1 diabetes, type 2
diabetes, type 3 diabetes, or a combination thereof.
13. The method of claim 9, wherein the myeloperoxidase related
pathology comprises leaky gut, endotoxemia, inflammatory bowel
disease or a combination thereof.
14. The method of claim 9, wherein the myeloperoxidase related
pathology comprises a dermatopic pathology including slow wound
healing, wrinkles, sun spots, and premature signs of aging.
15. The method of claim 9, wherein the treating of the subject is
prophylactic.
16. The method of claim 1, wherein the condition or disorder is
penile dysfunction.
17. The method of claim 1, wherein acutely raising of nitric oxide
levels in the subject enhances endothelial functioning, decreases
monocyte-mediated arterial plaque formation, decreases the
development of peripheral arterial disease, or a combination
thereof, wherein an increase or decreases refers to a level in the
subject prior to the administering of the therapeutically effective
combination.
18. The method of claim 1, wherein the subject is a human.
19. The method of claim 1, wherein the treating of the subject is
prophylactic.
20. The method of claim 1, wherein acutely raising nitric oxide
levels in the subject comprises raising salivary nitrite levels in
the subject beyond a level of the salivary nitrite in the subject
as compared to a level prior to administering the therapeutically
effective combination.
21. A system for acutely raising nitric oxide levels in a subject,
comprising: an effective amount of a NOS dependent source of nitric
oxide; an effective amount of a NOS independent source of nitric
oxide; and an effective amount of a myeloperoxidase inhibitor.
22. The system of claim 21, wherein at least one of the NOS
dependent source of nitric oxide, the NOS independent source of
nitric oxide, and the myeloperoxidase inhibitor are separate from
one another.
23. The system of claim 21, wherein at least one of the NOS
dependent source of nitric oxide, the NOS independent source of
nitric oxide, and the myeloperoxidase inhibitor are in separate
formulations.
24. The system of claim 21, wherein a level of nitric oxide in the
subject following administration of the composition is greater than
an amount provided by an equivalent amount of any one of the NOS
dependent source of nitric oxide, the NOS independent source of
nitric oxide, or the myeloperoxidase inhibitor.
25. The system of claim 24, wherein the level of nitric oxide is
greater than an additive amount of an equivalent amount of any one
of the NOS dependent source of nitric oxide, the NOS independent
source of nitric oxide, or the myeloperoxidase inhibitor.
26. The system of claim 21, wherein the system is formulated as a
kit.
Description
PRIORITY DATA
[0001] This application is a continuation of U.S. patent
application Ser. No. 15/756,548, filed Feb. 28, 2018, now issued as
U.S. Pat. No. 11,241,406, which claims the benefit of U.S.
Provisional Application Ser. No. 62/211,673, filed on Aug. 28,
2015, each of which is herein incorporated by reference.
BACKGROUND
[0002] Nitric oxide influences a number of metabolic pathways and
plays a role in vascular signal transduction, neuronal signal
transduction, smooth muscle contraction, bioenergetics, platelet
adhesion, platelet aggregation, immunity, and cell death. Reduced
bioavailable levels of nitric oxide are implicated in numerous
conditions and disorders. Nitric oxide plays a key role in the
functioning of the cardiovascular, nervous, pulmonary,
gastrointestinal, renal, and immune systems. Formulations and
methods that provide an acute increase in bioavailable NO would
provide a health benefit.
BRIEF DESCRIPTION OF THE DRAWINGS
[0003] Features and advantages of the invention will be apparent
from the detailed description that follows, and which taken in
conjunction with the accompanying figures together illustrate
features of the invention. It is understood that the figures merely
depict exemplary embodiments and are therefore, not to be
considered limiting in scope.
[0004] FIG. 1 schematically displays pathophysiologic conditions
related to poor eNOS-related NO biosynthesis;
[0005] FIG. 2 schematically displays the positive effects of NO in
peripheral vascular diseases related to cardiometaboic risk
factors;
[0006] FIG. 3 schematically displays NO production by NOS-dependent
(L-arginine) and NOS independent (NO3/NO2) pathways;
[0007] FIG. 4 schematically displays the nitric oxide cycle;
[0008] FIG. 5 schematically displays the pathways for NO2 reduction
to NO at specific sites of action addressing system
pathologies;
[0009] FIG. 6 schematically displays the multiple roles of
myeloperoxidase in cardiometabolic pathologies and oxidative
stress;
[0010] FIG. 7 schematically displays the dual role of
myeloperoxidase in acute disease prevention and chronic disease
causation;
[0011] FIGS. 8A and 8B depict the multifunctional role of
myeloperoxidase in the nitration of key proteins in cardiovascular
disease [8A] and the three putative sites of action inhibited
simultaneously by the novel phytocomplexes described by the instant
invention [8B];
[0012] FIGS. 9A and 9B graphically present the relative salivary NO
biomarker concentrations at 0, 30, 60, and 90 minutes post-dosing
for [9A] n=5 and n=4 subjects, respectively, for F1 and F2 and [9B]
a single individual consuming a double-dose of F1 and F2 on two
different occasions, in accordance with Examples 1 & 2; and
[0013] FIG. 10 graphically presents the relative salivary NO
biomarker concentrations at 0, 30, 60, and 90 minutes following
post-dosing of F3 for n=5 subjects in accordance with Example
3.
DETAILED DESCRIPTION
[0014] Before invention embodiments are disclosed and described, it
is to be understood that no limitation to the particular
structures, process steps, or materials disclosed herein is
intended, but also includes equivalents thereof as would be
recognized by those ordinarily skilled in the relevant arts. It
should also be understood that terminology employed herein is used
to describe particular examples only and is not intended to be
limiting. The same reference numerals in different drawings
represent the same element. Numbers provided in flow charts and
processes are provided for clarity in illustrating steps and
operations and do not necessarily indicate a particular order or
sequence. Unless defined otherwise, all technical and scientific
terms used herein have the same meaning as commonly understood by
one of ordinary skill in the art to which this disclosure belongs.
In the specification and the appended claims, the singular forms
include plural referents unless the context clearly dictates
otherwise.
[0015] As used in this written description, the singular forms "a,"
"an," and "the" specifically also provide express support for
plural referents, unless the content clearly dictates otherwise.
For example, "an excipient" provides support for one or more
excipients.
[0016] The term "about" is used herein refers to a degree of
deviation. It means approximately, in the region of, roughly, or
around. When the term "about" is used in conjunction with a
numerical range, it modifies that range by extending the boundaries
above and below the numerical values set forth. It is understood
that support in this specification for numerical values used in
connection with the term "about" is also provided for the exact
numerical value itself as though "about" were not used.
[0017] As used herein "acute" refers to obtaining the desired
effect within a set, or specifically intended, amount of time. In
one example, acute refers to an increase or decrease in the
concentration, production, secretion, biosynthesis, and/or
bioavailability occurring within 30-120 minutes post dosing and
lasting for up to 8 hours.
[0018] "Bergamot" refers to bergamot orange (Citrus bergamia
Risso). This citrus species, grows abundantly in the Calabria
region of southern Italy, and has been used in Calabrian folk
medicine to treat cardiovascular ailments for centuries. Bergamot
comprises two 3-hydroxymethylglutaryl (HMG) derivatives of
naturally occurring flavonoid glycosides brutieridin and melitidin.
These glycosides are the HMG derivatives of glucosylated hesperetin
and naringenin, respectively, and have a structural similarity to
the commercially available HMG-CoA reductase inhibitors known as
the statins. As used herein bergamot can be used interchangeably to
refer to the fruit and/or the extract.
[0019] As used herein "cardiometabolic-associated pathologies" or
"cardiometabolic risk factors" refers to any condition that
increases the risk of those pathologies associated with
cardiovascular dysfunction. This generally results from a
combination of decreasing the localized production of NO and
increasing MPO activity at the same site. A non-limiting example of
such pathologies, include: angina, arterial plaque buildup, deep
vein thrombosis, dementia, diabetes (types, 1, 2, and 3), diabetic
foot disorders, elevated glucose, insulin or HOMA score, elevated
hs-CRP (levels greater than 1.0 pmol/L), elevated myeloperoxidase
(levels greater than 350 pmol/L), endothelial dysfunction, erectile
dysfunction, fibrinogen levels greater than 370 pmol/L, HDL
modification, heart attack, heart failure, hypertension (BP greater
than 140/90), lipoprotein-associated phospholipase A2 (LpPLA2
levels greater than 200 pmol/L), macular degeneration,
monocyte-mediated arterial plaque formation, oxidation of LDL,
peridontal disease, peripheral arterial disease, platelet
stickiness, portal hypertension, pregnancy/pre-eclampsia, pulmonary
hypertension, renal failure, serum low density lipoprotein (LDL)
greater than 150 mg/dL, serum triglycerides greater than 150 mg/dL,
sleep apnea, smooth muscle cell proliferation, stroke, and
vasculitis.
[0020] As used herein, chemical compounds or simply "compounds" may
be identified either by their chemical structure, chemical name, or
common name. In the event that the chemical structure, chemical
name, or common name conflict, the chemical structure is
determinative of the identity of the compound. The compounds
described herein may contain one or more chiral centers and/or
double bonds and therefore, may exist as stereoisomers, such as
double-bond isomers (i.e., geometric isomers), enantiomers, or
diastereomers. Accordingly, the chemical structures depicted herein
encompass all possible enantiomers and stereoisomers of the
illustrated or identified compounds including the
stereoisomerically pure form (e.g., geometrically pure,
enantiomerically pure, or diastereomerically pure) and enantiomeric
and stereoisomeric mixtures. Enantiomeric and stereoisomeric
mixtures can be resolved into their component enantiomers or
stereoisomers using separation techniques or chiral synthesis
techniques well known to the skilled artisan. The compounds may
also exist in several tautomeric forms including the enol form, the
keto form and mixtures thereof. Accordingly, the chemical
structures encompass all possible tautomeric forms of the
illustrated or identified compounds. The compounds described also
encompass isotopically labeled compounds where one or more atoms
have an atomic mass different from the atomic mass conventionally
found in nature. Examples of isotopes that may be incorporated into
the compounds of the invention include, but are not limited to,
.sup.2H, .sup.3H, .sup.13C, .sup.14C, .sup.15N, .sup.18O, .sup.17O,
etc. Compounds may exist in unsolvated forms as well as solvated
forms, including hydrated forms and as N-oxides. In general,
compounds may be hydrated, solvated, or N-oxides. Certain compounds
may exist in multiple crystalline or amorphous forms. Also
contemplated are congeners, analogs, hydrolysis products,
metabolites, and precursor or prodrugs of the compound. In general,
all physical forms are equivalent for the uses contemplated herein
and are intended to be within the scope of the present
disclosure.
[0021] Concentrations, amounts, and other numerical data may be
expressed or presented herein in a range format. It is to be
understood that such a range format is used merely for convenience
and brevity and thus should be interpreted flexibly to include not
only the numerical values explicitly recited as the limits or
endpoints of the range, but also to include all the individual
numerical values and/or sub-ranges encompassed within that range as
if each numerical value (including fractions) and sub-range is
explicitly recited. As an illustration, a numerical range of "about
1 to about 5" should be interpreted to include not only the
explicitly recited values of about 1 to about 5, but also include
individual values and sub-ranges within the indicated range. Thus,
included in this numerical range are individual values such as 2,
2.6, 3, 3.8, and 4 and sub-ranges such as from 1-3, from 2-4, and
from 3-5, etc., as well as 1, 2, 3, 4, and 5, individually.
[0022] As used herein a "concentrate" refers to an extract of a
source that contains at least the same amount of active fractions,
compounds, or other constituents, in a smaller volume than in the
source itself. In one example, a "concentrate" may be a dried
powder derived from a component that does not include the use of
any solvents during the concentration process.
[0023] Comparative terms such as "more effectively," "greater
than," "improved," "enhanced," and like terms can be used to state
a result achieved or property present in a formulation or process
that has a measurably better or more positive outcome than the
thing to which comparison is made. In some instances comparison may
be made to the prior art.
[0024] As used herein, "comprises," "comprising," "containing," and
"having" and the like can have the meaning ascribed to them in U.S.
Patent law and can mean "includes," "including," and the like, and
are generally interpreted to be open ended terms. The terms
"consisting of" or "consists of" are closed terms, and include only
the components, structures, steps, or the like specifically listed
in conjunction with such terms, as well as that which is in
accordance with U.S. Patent law. "Consisting essentially of" or
"consists essentially of" have the meaning generally ascribed to
them by U.S. Patent law. In particular, such terms are generally
closed terms, with the exception of allowing inclusion of
additional items, materials, components, steps, or elements, that
do not materially affect the basic and novel characteristics or
function of the item(s) used in connection therewith. For example,
trace elements present in a composition, but not affecting the
composition's nature or characteristics would be permissible if
present under the "consisting essentially of" language, even though
not expressly recited in a list of items following such
terminology. When using an open ended term in the written
description, like "comprising" or "including," it is understood
that direct support should be afforded also to "consisting
essentially of" language, as well as, "consisting of" language as
if stated explicitly and vice versa.
[0025] As used herein, a "derivative" is a compound obtained from a
source compound an analog, homolog tautomeric form, stereoisomer,
polymorph, hydrate, pharmaceutically acceptable salt or
pharmaceutically acceptable solvate thereof, by a simple chemical
process converting one or more functional groups, by means of
oxidation, hydrogenation, alkylation, esterification, halogenation
and the like. The term "analog" refers to a compound having a
structure similar to that of another one, but differing from it
with respect to a certain component. The compound may differ in one
or more atoms, functional groups, or substructures, which may be
replaced with other atoms, groups, or substructures. In one aspect,
such structures possess at least the same or a similar therapeutic
efficacy for a given indication. The term "tautomer" or "tautomeric
form" refers to structural isomers of different energies which are
interconvertible via a low energy barrier. The term "stereoisomer"
refers to one of a set of isomers whose molecules have the same
number and kind of atoms bonded to each other, but which differ in
the way these atoms are arranged in space. The term "polymorph"
refers to crystallographically distinct forms of a substance. In
addition, an agent can be said to be "derived" from a source
containing many compounds or agents, such as a plant, fungus,
bacteria, or other organism. In this context, the agent can be
described or otherwise referred to in terms of its source, rather
than by its own properties, characteristics, name, or attributes
per se. For example, an extract obtained from a plant may be
described as "derived" from the plant.
[0026] The term "dosage unit" is understood to mean a unitary, i.e.
a single dose which is capable of being administered to a subject
or patient, and that may be readily handled and packed, remaining
as a physically and chemically stable unit dose comprising either
the active ingredient as such or a mixture of it with solid or
liquid pharmaceutical vehicle materials.
[0027] The phrase "effective amount," "therapeutically effective
amount," or "therapeutically effective rate(s)" of an active
ingredient refers to a non-toxic, but sufficient amount or delivery
rates of the active ingredient, to achieve therapeutic results in
treating a disease or condition for which the drug is being
delivered. It is understood that various biological factors may
affect the ability of a substance to perform its intended task.
Therefore, an "effective amount," "therapeutically effective
amount," or "therapeutically effective rate(s)" may be dependent in
some instances on such biological factors. Further, while the
achievement of therapeutic effects may be measured by a physician
or other qualified medical personnel using evaluations known in the
art, it is recognized that individual variation and response to
treatments may make the achievement of therapeutic effects a
subjective decision. The determination of a therapeutically
effective amount or delivery rate is well within the ordinary skill
in the art of pharmaceutical sciences and medicine.
[0028] The term "extract" refers to those substances prepared using
a solvent, e.g., ethanol, water, steam, superheated water,
methanol, hexane, chloroform liquid, liquid CO.sub.2, liquid
N.sub.2, propane, supercritical CO.sub.2, or any combination
thereof. Extracts, as used herein, can refer to an extract in a
liquid form, or can refer to a product obtained from further
processing of the liquid form, such as a dried powder or other
solid form. Extracts may take many forms including but not limited
to: solid, liquid, particulate, chopped, distillate, etc. and may
be performed by any number of procedures or protocols, such as
chopping, grinding, pulverizing, boiling, steaming, soaking,
steeping, infusing, applying a gas, etc., and may employ any
suitable reagents, such as water, alcohol, steam, or other organic
materials. Extracts typically have a given purity percentage and
can be relatively to highly pure. In some embodiments, extracts can
be phytoextracts made from specific parts of a source, such as the
skin, pulp, leaves, flowers, fruits of a plant etc., or can be made
from the whole source. In some aspects an extract may include one
or more active fractions or active agents. In some extracts,
maltodextrin can be added as a carrier. In some aspects, the purity
of an extract can be controlled by, or be a function of the
extraction process or protocol.
[0029] As used herein, "formulation" and "composition" can be used
interchangeably and refer to a combination of at least two
ingredients. In some embodiments, at least one ingredient may be an
active agent or otherwise have properties that exert physiologic
activity when administered to a subject.
[0030] Formulation or compositional ingredients included or recited
herein are to be presumed to be in wt % unless specifically stated
otherwise. In addition, ingredient amounts presented in the form of
ratios are to be presumed to be in wt % (e.g. % w/w) ratios. As
such, a composition containing four ingredients at a 1:1:1:1 ratio
would indicate that each ingredient is present in an amount of 25
wt %. Accordingly, in some aspects, the amount of an ingredient in
a composition or formulation in terms of wt % can be derived from a
numerical ratio value.
[0031] As used herein, an "increase" or a "decrease" in
concentrations or levels means a change of at least 5%.
[0032] As used herein, "inhibit," "inhibiting," "inhibition," and
like terms refer to the act of reducing, minimizing, stopping or
arresting a function, role, or activity. For example, these terms
can mean reducing, minimizing, stopping, arresting, or effectively
reducing, minimizing, stopping, or arresting myeloperoxidase
activity in a subject.
[0033] As used herein, "Leaky Gut Syndrome (LGS)" is an increase in
permeability of the intestinal mucosa to luminal macromolecules,
antigens and toxins associated with inflammatory degenerative
and/or atrophic mucosal damage. LGS can lead to any number of
seemingly unrelated symptoms affecting every organ system in the
body. LGS has also been linked with having a causative role in a
large number of distinct illnesses. Many of these are autoimmune
diseases, which means the immune system attacks the body's own
cells. LGS plays a role in these types of illness because it
increases immune reactions to food particles and then cross
reactivity may occur meaning that the immune system attacks body
tissues that are chemically similar to the foods to which it has
become sensitized. A sampling of the many diseases in which leaky
gut syndrome may have a role includes: rheumatoid arthritis,
osteoarthritis, asthma, multiple sclerosis, vasculitis, Crohn's
Disease, colitis, Addison's disease, lupus, thyroiditis, chronic
fatigue syndrome, and fibromyalgia.
[0034] As used herein, "linear inhibitory effect" or
"dose-response" refers to a linear decrease in secretion or
biosynthesis resulting from all concentrations of the inhibiting
material over a dose-response curve. For example, inhibition at low
concentrations followed by a failure of inhibition or increased
secretion at higher concentrations represents a lack of a linear
inhibitory effect.
[0035] The term "or" is used in the "inclusive" sense of "and/or"
and not the "exclusive" sense of "either/or."
[0036] As used herein, "pharmaceutically acceptable" refers
generally to materials which are suitable for administration to a
subject in connection with an active agent or ingredient. For
example, a "pharmaceutically acceptable carrier" can be any
substance or material that can be suitably combined with an active
agent to provide a composition or formulation suitable for
administration to a subject. Excipients, diluents, and other
ingredients used in or used to prepare a formulation or composition
for administration to a subject can be used with such term.
[0037] The term "prevent" and its variants refer to prophylaxis
against a particular undesirable physiological condition. The
prophylaxis may be partial or complete. Partial prophylaxis may
result in the delayed onset of a physiological condition. The
person skilled in the art will recognize the desirability of
delaying onset of a physiological condition, and will know to
administer the compositions of the invention to subjects who are at
risk for certain physiological conditions in order to delay the
onset of those conditions. For example, the person skilled in the
art will recognize that obese subjects are at elevated risk for
coronary artery disease. Thus, the person skilled in the art will
administer compositions of the invention in order to increase
insulin sensitivity in an obese, whereby the onset of diabetes
mellitus or dyslipemia may be prevented entirely or delayed.
[0038] As used herein, a "subject" refers to an organism that
produces nitric oxide and myloperoxidase in the course of its
cellular function. In one aspect, a subject can be a mammal. In
another aspect, a subject can be a human. In another aspect, the
subject can be a domesticated animal or livestock.
[0039] As used herein, "substantial" or "substantially" when used
in reference to a quantity or amount of a material, or a specific
characteristic thereof, refers to an amount that is sufficient to
provide an effect that the material or characteristic was intended
to provide. The exact degree of deviation allowable may in some
cases depend on the specific context. Similarly, "substantially
free of" or the like refers to the lack of an identified element or
agent in a composition. Particularly, elements that are identified
as being "substantially free of" are either completely absent from
the composition, or are included only in amounts which are small
enough so as to have no measurable effect on the composition.
[0040] The terms "treat," "treating," or "treatment" as used herein
and as well understood in the art, mean an approach for obtaining
beneficial or desired results, including without limitation
clinical results in a subject being treated. Beneficial or desired
results can include, but are not limited to, alleviation or
amelioration of one or more signs or symptoms of a condition,
diminishment of extent of disease, stabilizing (i.e. not worsening)
the state of a disease or condition, delaying or slowing of disease
progression, amelioration or palliation of the disease state,
diminishment of the reoccurrence of disease, and remission (whether
partial or total), whether detectable or undetectable. "Treat,"
"treating" and "treatment" can also mean prolonging survival as
compared to expected survival if not receiving treatment and can be
prophylactic. Such prophylactic treatment can also be referred to
as prevention or prophylaxis of a disease or condition. The
prophylaxis may be partial or complete. Partial prophylaxis may
result in the delayed onset of a physiological condition.
[0041] As used herein, the term "solvent" refers to a liquid of
gaseous, aqueous, or organic nature possessing the necessary
characteristics to extract solid material from a plant product.
Examples of solvents would include, but not limited to, water,
steam, superheated water, methanol, ethanol, ethyl acetate, hexane,
chloroform, liquid CO.sub.2, liquid N.sub.2, propane, or any
combinations of such materials.
[0042] As used herein, "synergistic" means more than the additive
effect of the individual components against a mechanism of action.
For example if F1 produces response X, F2 produces response Y, then
the combination of F1+F2>X+Y. In some situations F2 produces no
response and the value for Y is equal to zero.
[0043] As used herein, a plurality of items, structural elements,
compositional elements, and/or materials may be presented in a
common list for convenience. However, these lists should be
construed as though each member of the list is individually
identified as a separate and unique member. Thus, no individual
member of such list should be construed as a de facto equivalent of
any other member of the same list solely based on their
presentation in a common group without indications to the
contrary.
[0044] Any steps recited in any method or process claims may be
executed in any order and are not limited to the order presented in
the claims unless otherwise stated.
[0045] Nitric Oxide (NO) is a free radical, actively produced in
the human body. NO plays a role in the normal functioning of the
cardiovascular, nervous, pulmonary, gastrointestinal, renal, and
immune systems. See Table 1.
TABLE-US-00001 TABLE 1 Exemplary Established Functions of Nitric
Oxide System Function Cardiovascular Controls vascular tone.
Relaxes vascular smooth muscles and reduces blood pressure. Dilates
vessels and relieves the pain of angina. Inhibits the aggregation
of platelets within the vessels and prevents thrombotic events.
Controls vascular tone. Relaxes vascular smooth muscles and reduces
blood pressure. Dilates vessels and relieves the pain of angina.
Inhibits the aggregation of platelets within the vessels and
prevents thrombotic events. Nervous Acts as a neurotransmitter,
including in the autonomic nervous system. Increases cerebral blood
flow and oxygenation to the brain. Important mediator in penile
erection during sexual arousal. Pulmonary Dilates pulmonary
vessels. Benefits Adult Respiratory Distress Syndrome, pulmonary
hypertension and Chronic Obstructive Airway Disease. Produced in
abnormal amounts in inflammatory lung conditions. Concentration of
NO in exhaled air can be taken as a marker of airway inflammation.
Gastrointestinal Regulates the relaxation of smooth muscles.
Controls peristalsis and the function of sphincters. Renal
Increases blood flow to the kidney due to its vasodilatory effect.
Increases the glomerular filtration rate and the production of
urine. Immune Modulates T cell-mediated immune response.
[0046] Defective control of nitric oxide levels can play a role in
numerous pathologies as shown in FIG. 1. These pathologies include:
angina, Alzheimer's disease, dementia, diabetic foot, diabetes,
erectile dysfunction, heart attack, heart failure, hypertension,
portal hypertension, peripheral arterial disease, pulmonary
hypertension, macular degeneration, periodontal disease, pregnancy
related pre-eclampsia, Raynaud's disease, renal failure, sleep
apnea, and stroke. With respect to cardiovascular pathophysiology,
nitric oxide can activate soluble guanylate cyclase (sGC)-cGMP
signal transduction pathways which can mediate various beneficial
physiological effects in the cardiovascular system including
vasodilation, inhibition of platelet aggregation, reduction of
monocyte "stickiness" to prevent formation of plaque, reduction of
smooth muscle cell proliferation, inhibition of superoxide radical
formation, and reduction of LDL oxidation. See FIG. 2.
[0047] Reference is made hereinafter in detail to specific
embodiments of the invention. While the invention will be described
in conjunction with these specific embodiments, it will be
understood that it is not intended to limit the invention to such
specific embodiments. On the contrary, it is intended to cover
alternatives, modifications, and equivalents as may be included
within the spirit and scope of the invention as defined by the
appended claims. In the following description, numerous specific
details are set forth in order to provide a thorough understanding
of the present invention. The present invention may be practiced
without some or all of these specific details. In other instances,
well known process operations have not been described in detail, in
order not to unnecessarily obscure the present invention.
[0048] The present disclosure relates to compositions and methods
for acutely raising nitric oxide levels in a subject. The acute
raising of nitric oxide levels can be beneficial to the health of a
subject. The compositions can include: an effective amount of a NOS
(nitric oxide synthase) dependent source of nitric oxide, an
effective amount of a NOS independent source of nitric oxide, and a
myeloperoxidase (MPO) inhibitor. When administered to a subject,
the composition can acutely raise nitric oxide levels in the
subject above the level of nitric oxide that is available in the
subject prior to the administration of the composition. The NOS
dependent source of NO and the NOS independent source of NO can
function to increase NO levels in the subject, see FIG. 3, while
the MPO inhibitor can prevent oxidative stress associated with MPO.
In one example, the composition can acutely raise the level of
nitric oxide in the subject following administration of the
composition to an amount that is greater than the amount provided
by an equivalent amount of any one of the NOS dependent source of
nitric oxide, the NOS independent source of nitric oxide, or the
myeloperoxidase inhibitor.
[0049] NOS dependent sources of nitric oxide (NO) can include
compounds that can be catalyzed by NOS to produce NO. The nitric
oxide cycle is shown in FIG. 4. As shown in FIG. 4 L-arginine,
L-citrulline, and ornithine are all components in the nitric oxide
cycle. Nitric oxide produced from NOS dependent sources have a
half-life of approximately one second and are quickly oxidized to
NO.sub.3/NO.sub.2 or react with thiols or amines in the body. In
one example, the NOS dependent source of nitric oxide in the
composition, can include L-arginine, L-citrulline, ornithine, or a
combination thereof. In one example, the NOS dependent sources of
nitric oxide can comprise from about 40 wt % to about 98 wt % of
the composition. In another example, the NOS dependent sources of
nitric oxide can comprise from 0.01 wt % to 15 wt % of the
composition.
[0050] In another example, the NOS dependent source of nitric oxide
comprises L-arginine. In one example, the L-arginine in the
composition can range from about 1 wt % to about 80 wt %. In
another example, the L-arginine in the composition can range from
about 0.5 wt % to about 90 wt %. In yet another example, the
L-arginine in the composition can range from about 0.25 wt % to
about 95 wt %. In a further example, the L-arginine can be in the
composition from about 0.125 wt % to about 99 wt %. In yet a
further example, the L-arginine can be in the composition from
about 40 wt % to about 95 wt %. In another example, the L-arginine
can be in the composition from 0 wt % to about 10 wt %.
[0051] In another example, the NOS dependent source of nitric oxide
can include L-citrulline. In one example, the L-citrulline can be
present in the composition from about 1 wt % to about 80 wt %. In
another example, the L-citrulline can be present in the composition
from about 0.5 wt % to about 90 wt %. In yet another example, the
L-citrulline can be present in the composition from about 0.25 wt %
to about 95 wt %. In a further example, the L-citrulline can be
present in the composition from about 0.125 wt % to about 99 wt %.
In yet a further example, the L-citrulline can be present in the
composition from about 0.1 wt % to about 5 wt %. In yet another
example, the L-citrulline can be present from about 0.5 wt % to
about 2.5 wt %.
[0052] In some examples, the source of the L-citrulline can be a
watermelon extract. In one example, the watermelon extract can
include about 20 wt % citrulline. In another example, the
watermelon extract can include about 10 wt % citrulline, about 15
wt % citrulline, about 25 wt % citrulline, about 30 wt %
citrulline, about 40 wt % citrulline, or about 50 wt %
citrulline.
[0053] In yet another example, the NOS dependent source of nitric
oxide can include L-arginine and L-citrulline. In one example, the
L-arginine can range from about 1 wt % to about 80 wt % and the
L-citrulline can range from about 1 wt % to about 80 wt %. In
another example, the L-arginine can range from about 0.5 wt % to
about 90 wt % in and the L-citrulline can range from about 0.5 wt %
to about 90 wt %. In yet another example, the L-arginine can range
from about 0.25 wt % to about 95 wt % and the L-citrulline can
range from about 0.25 wt % to about 95 wt %. In yet a further
example, the L-arginine can range from about 40 wt % to about 95 wt
% and the L-citrulline can range from about 0.1 wt % to about 5 wt
%.
[0054] The ratio of the weight percentage of L-arginine and
L-citrulline can also vary. In one example, the L-arginine and
L-citrulline can be present in the composition at a weight percent
concentration ranging from about 1:5 to about 5:1, respectively. In
another example, the L-arginine and L-citrulline can be present in
the composition at a weight percent concentration ranging from
about 1:1 to about 5:1, respectively. In yet another example, the
L-arginine and L-citrulline can be present in the composition at a
weight percent concentration of about 2:1, respectively.
[0055] Turing now to the NOS independent source of nitric oxide.
NOS independent sources of nitric oxide, can include dietary
sources of nitrates/nitrites. NO.sub.3 (nitrate) can be reduced by
facultative anaerobic bacteria on the dorsal surface of the tongue
to nitrite (NO.sub.2). The NO.sub.2 can then be enzymatically
reduced to NO. See FIG. 5. The bioavailability of dietary NO.sub.3
is 100%. Orally consumed NO.sub.3 can reach peak plasma
concentration in about 1 hour and has a half-life in plasma of
about 5 hours.
[0056] In one example, the NOS independent source of nitric oxide
can include a member selected from the group consisting of beet
root extract, vitamin B1, collard green extract, nut powders,
spinach extract, broccoli extract, lettuce extract, celery, kale,
watercress, carrot, arugula, mustard greens, or a combination
thereof.
[0057] In one example, the NOS dependent source of nitric oxide can
be in the composition at a range from about 1 wt % to about 80 wt
%. In another example, the NOS dependent source of nitric oxide can
be in the composition at a range from about 0.01 wt % to about 90
wt %. In another example, the NOS dependent source of nitric oxide
can be present in the composition from about 15 wt % to about 30 wt
%.
[0058] In one example, the NOS independent source of nitric oxide
can include beet root extract, vitamin B1, or a combination
thereof. In another example, the NOS independent source of nitric
oxide can include the beet root extract and the vitamin B1. In one
example, including beet root extract and vitamin B1, the beet root
extract can range from about 5 wt % to about 90 wt % and the
vitamin B1 can range from about 0.01 wt % to about 80 wt % of the
composition. In another example, including beet root extract and
vitamin B1, the beet root extract can range from about 1 wt % to
about 95 wt % and the vitamin B1 can range from about 0.005 wt % to
about 99 wt % of the composition. In yet another example, including
beet root extract and vitamin B1, the beet root extract can range
from about 0.1 wt % to about 99 wt % and the vitamin B1 can range
from about 0.0025 wt % to about 0.995 wt % of the composition. In a
further example, the beet root extract can range from about 15 wt %
to about 30 wt % and the vitamin B1 can range from about 0.5 wt %
to about 5 wt %.
[0059] In formulations including beet root extract and vitamin B1,
the ratio of these components can also vary. In one example, the
beet root extract and vitamin B1 can be present in the composition
at a weight percent concentration ratio ranging from about 50:1 to
about 10:1, respectively. In another example, the beet root extract
and vitamin B1 can be present in the composition at a weight
percent concentration ratio ranging from about 100:1 to about 1:1,
respectively. In a further example, the beet root extract and
vitamin B1 can be present in the composition at a weight percent
concentration ratio ranging from about 1,000:1 to about 1:2,
respectively.
[0060] Turning now to the myeloperoxidase inhibitor.
Myeloperoxidase (MPO) is a heme containing enzyme that produces
hypochlorous acid (HOCl) as part of the body's defense mechanism on
invading organisms. See FIGS. 6 and 7. MPO also exacerbates
inflammatory diseases and promotes oxidative stress. Accordingly,
MPO inhibitors can be useful in the treatment of conditions and
pathologies associated with oxidative stress.
[0061] In one example, the myeloperoxidase inhibitor can be
pomegranate fruit extract, red grape polyphenols, apple extract,
blueberry extract, capsicum extract, grape extract, green tea
extract, olive extract, bergamot extract, mangosteen extract,
mangosteen fruit, or a combination thereof. In one example, the
myeloperoxidase inhibitor can range from about 1 wt % to about 90
wt %. In another example, the myeloperoxidase inhibitor can range
from about 0.5 wt % to about 95 wt %. In yet another example, the
myeloperoxidase inhibitor can range from about 0.25 wt % to about
99 wt %. In another example, the myeloperoxidase inhibitor can
range from about 1 wt % to about 15 wt %. In yet a further example,
the myeloperoxidase inhibitor can range from about 3 wt % to about
12 wt %.
[0062] In one example, the myeloperoxidase inhibitor can include
pomegranate fruit extract, red grape polyphenols, or a combination
thereof. In another example, of the composition, the
myeloperoxidase inhibitor can include pomegranate fruit extract and
red grape polyphenols. In one example, the pomegranate fruit
extract can range from about 1 wt % to about 80 wt % and the red
grape polyphenols can range from about 1 wt % to about 80 wt % of
the composition. In another example, the pomegranate fruit extract
can range from about 0.5 wt % to about 90 wt % and the red grape
polyphenols can range from about 0.5 wt % to about 90 wt % of the
composition. In yet another example, the pomegranate fruit extract
can range from about 0.25 wt % to about 95 wt % and the red grape
polyphenols can range from about 0.25 wt % to about 95 wt % of the
composition. In one example, the pomegranate fruit extract can
range from about 0.25 wt % to about 10 wt % and the red grape
polyphenols can range from about 0.01 wt % to about 2 wt % of the
composition.
[0063] In another example, the myeloperoxidase inhibitor can
include apple extract, grape extract, green tea extract, and olive
extract. In one example, the apple extract, the grape extract, the
green tea extract, and the olive extract collectively can range
from about 1 wt % to about 80 wt % of the composition. In another
example, the apple extract can range from about 0.01 wt % to about
80 wt %, the grape extract can range from about 0.01 wt % to about
80 wt %, the green tea extract can range from about 0.01 wt % to
about 80 wt %, and the olive extract can range from about 0.01 wt %
to about 80 wt % of the claimed composition. In yet another
example, the apple extract can range from about 0.01 wt % to about
5 wt %, the grape extract can range from about 0.01 wt % to about 7
wt %, the green tea extract can range from about 0 wt % to about 1
wt %, and the olive extract can range from about 0 wt % to about 1
wt % of the claimed composition. In another example, the apple
extract, the grape extract, the green tea extract, and the green
tea leaf extract, are each present in the composition from about
0.01 wt % to about 1 wt %.
[0064] In one example, the apple extract can comprise an extract
derived from a member selected from the group consisting of Malus
domestica, Malta sieversii, Malta sylvestris, Malus pumila, and
combinations thereof. In one example the apple extract can be
derived from the species Malus pumila. In one example, the apple
extract can be derived from a combination of Malus domestica and
Malus pumila. In some embodiments the apple extract can comprise
any or all parts of the apple, including but not limited to the
skin, flesh/fruit (exocarp, mesocarp, and/or endocarp), seed,
stalk, stem, leaf, or a combination thereof. In one example, the
apple extract comprises the skin and fruit of the apple. In some
embodiments, the extract can be derived from immature apples. In
one embodiment, an extraction solvent can be ethanol.
[0065] In one example, the grape extract can comprise a member
selected from the group consisting of Vitis vinifera, Vitis
labrusca, Vitis riparia, Vitis rotundifolia, Vitis rupestris, Vitis
aestivalis, Vitis mustangensis, and combinations thereof. In one
example, the grape extract can be derived from Vitis vinifera. In
some embodiments, the grape extract can comprise any or all parts
of the grape including but not limited to the skin, flesh/fruit,
seed, vascular bundles, vine, leaves, or combination thereof. In
one embodiment, the grape extract can be derived from the seeds. In
another embodiment, the grape extract can be derived from the skin.
In yet another embodiment, the grape extract can be derived from
the seeds and skin of the grape. In some embodiments, the grape
extract comprises from about 75 wt % to about 95 wt % phenolics on
a dry weight basis. In other embodiments, the grape extract can
comprise from about 80 wt % to 97 wt % phenolics on a dry weight
basis. In one example, the extraction solvent can be ethanol,
water, or a mixture thereof.
[0066] In one example, the green tea extract can be derived from
Camellia sinensis. In some embodiments, the green tea extract can
comprise any or all parts of the tea including but not limited to
the leaf, seed, stem, flower, or combination thereof. In one
embodiment, the green tea extract can be derived from the leaves.
In another example, the extract solvent can be water, ethanol,
ethyl acetate, or combinations thereof.
[0067] In one example, the olive extract comprises a subspecies of
Olea europea selected from the group consisting of the subspecies
europea, cuspidiata, guanchica, cerasiformis, maroccana,
laperrinei, cerasiformis, or a combination thereof. In some
embodiments, the olive extract can comprise any or all parts of the
olive including but not limited to the leaf, seed, pulp, fruit,
stem, or combination thereof. In one embodiment, the olive extract
can be derived from the leaves. In another example, the extraction
solvent can be an ethanol and water solution.
[0068] In some embodiments, the plant or herb to extract ratio can
range from about 1 to about 10. In other examples, the raw plant or
herb to extract ratio can be from about 2 to about 5, from about 4
to about 7, or from about 8 to about 10.
[0069] By way of example, in some embodiments, each extract can be
present at a ratio of from about 1 to about 50 times the amount of
another extract. In one aspect, the apple extract can be present in
the formulation at a ratio of from 1 to 50 times the amount of a
grape, green tea, or olive extract. In another aspect, the apple
extract can be present in the formulation at a ratio of from about
1 to 25 times the amount of a grape, green tea, or olive extract.
In a further aspect, the apple extract in the formulation can be
present at a ratio of from 1 to 10 times the amount of a grape,
green tea, or olive extract. In an additional aspect, the apple
extract can be present in at a ratio of from 1 to 5 times the
amount of a grape, green tea, or olive extract. In yet another
aspect, the apple extract can be present in the formulation at a
ratio of 1 times the amount of a grape, green tea, or olive
extract.
[0070] Any specific numerical value within the numerical range is
included. In fact, each of the apple, grape, green tea, and olive
extracts may be present in a ratio of anywhere between 1 to 50
times and 1 times the amount of the other extracts. For example,
the amount of apple extract to grape extract to green tea extract
to olive extract may in some embodiments be 1-25:1-25:1-25:1-25
respectively. As such, any number given specific ratio that yields
a synergistic effect as recited herein can be used, for example
25:1:1:1, 1:25:1:1, 1:1:25:1, or 1:1:1:25. When considered in terms
of wt %, this would equate to one ingredient being present in an
amount of 89.28 wt % and the other three ingredients being present
in amounts of 3.57 wt %. This can be considered either in terms of
the formulation as a whole, or in terms of the myeloperoxidase
inhibitor in the formulation only. For example, these four
extracts, at a 1:1:1:1 ratio would result in a relative amount of
each at 25 wt % each (i.e. 100/4=25). In one example, the apple
extract, grape extract, green tea extract, and olive extract can be
present in the composition at a weight ratio of about 1:1:1:1. In
another example, the apple extract, grape extract, green tea
extract, and olive extract can be present in the composition at a
weight ratio of about 6:1:3:1. In a further example, at least one
of the apple extract, the grape extract, the green tea extract, and
the olive extract are present in the composition in a different
amount.
[0071] In one example, the myeloperoxidase inhibitor can further
include pomegranate fruit extract. In another example, the
myeloperoxidase inhibitor can further include blueberry fruit
extract, capsicum fruit extract, and turmeric root extract. In one
example, the blueberry extract/concentrate can be obtained from
Vaccinium angustifolium. In one example the blueberry concentrate
can be a dried powder created without the use of a solvent. In one
embodiment, it can take about 5 kg, about 8 kg, about 10 kg, or
about 12 kg of blueberries to obtain 1 kg of dried powder. In one
embodiment, the capsicum extract can be obtained from Capsicum
annuum. In some embodiments, capsicum extract can be derived from
powdered dried ripe fruits. In one example the turmeric extract can
be obtained from Curcuma longa. In some embodiments, the turmeric
extract can be derived from a turmeric powder. In on embodiment,
the turmeric powder can have from about 1 to about 10%
curcuminoids, from about 3 to about 5% curcuminoids, from about 2%
to about 8% curcuminoids, or from about 4% to about 12%
curcuminoids.
[0072] The amount of the myeloperoxidase inhibitor can also vary.
In one example, the blueberry fruit extract can range from about
0.01 wt % to about 80 wt %, the capsicum fruit extract can range
from about 0.01 wt % to about 80 wt %, and the turmeric root
extract can range from about 0.01 wt % to about 80 wt % of the
composition. In another example, the blueberry fruit extract can
range from about 0.005 wt % to about 90 wt %, the capsicum fruit
extract can range from about 0.005 wt % to about 90 wt %, and the
turmeric root extract can range from about 0.005 wt % to about 90
wt % of the composition. In yet another example, the blueberry
fruit extract, the capsicum fruit extract, and the turmeric root
extract can each range from about 0.005 wt % to about 5 wt % of the
composition. In a further example, the blueberry fruit extract, the
capsicum fruit extract, and the turmeric root extract can each
range from about 0.1 wt % to about 2.5 wt % of the composition.
[0073] When present, in one example, the apple extract, the grape
extract, the green tea extract, the olive extract, the blueberry
fruit extract, the capsicum fruit extract, and the turmeric root
extract can be present in the composition at a weight ratio of
about 1:1:1:1:1:1:1. In another example, at least one of the
extracts can be present in a different amount.
[0074] In yet a further example, the myeloperoxidase inhibitor can
further include turmeric rhizome extract and mangosteen. The
mangosteen can include a Garcinia mangostana and can be in the form
of a fruit, a fruit extract, a pericarp extract, or a combination
thereof. In one example, the mangosteen can include a mangosteen
fruit extract, a mangosteen pericarp extract, or a combination
thereof. In one example, the turmeric rhizome extract can range
from about 0.01 wt % to about 80 wt % and the mangosteen can range
from about 0.01 wt % to about 80 wt % of the composition. In
another example, the turmeric rhizome extract can range from about
0.005 wt % to about 90 wt % and the mangosteen can range from about
0.005 wt % to about 90 wt % of the composition. In yet another
example, the turmeric rhizome extract can range from about 0.0025
wt % to about 95 wt % and the mangosteen can range from about
0.0025 wt % to about 95 wt % of the composition. In yet another
example, the turmeric rhizome extract and the mangosteen can each
range from about 0.005 wt % to about 5 wt % of the composition. In
one example, the apple extract, the grape extract, the green tea
extract, the olive extract, the blueberry fruit extract, the
capsicum fruit extract, the turmeric root extract, the turmeric
rhizome extract, and the mangosteen can be present in the
composition at a weight ratio of about 1:1:1:1:1:1:1:1:1. In
another example, at least one of the extracts can be present at a
different amount than the other extracts.
[0075] In one example, the myeloperoxidase inhibitor can further
include bergamot fruit extract. In one example, the bergamot fruit
extract can be derived from Citrus bergamia Risso. In another
example, the bergamot fruit extract can range from about 0.01 wt %
to about 80 wt % of the composition. In yet another example, the
bergamot fruit extract can range from about 0.005 wt % to about 90
wt % of the composition. In a further example, the bergamot fruit
extract can range from about 0.0025 wt % to about 90 wt % of the
composition. In yet another example, the bergamot fruit extract can
comprise from about 0.0025 wt % to about 5 wt % of the composition.
In one example, the apple extract, the grape extract, the green tea
extract, the olive extract, the blueberry fruit extract, the
capsicum fruit extract, the turmeric root extract, the turmeric
rhizome extract, the mangosteen, and the bergamot fruit extract can
be present in the composition at a weight ratio of about
1:1:1:1:1:1:1:1:1:1.
[0076] In yet another example, the myeloperoxidase inhibitor can
include sodium copper chlorophyllin, spearmint (Mentha spicada)
oil, or a combination thereof. In one example, the sodium copper
chlorophyllin can be present in the composition from about 10 wt %
to about 80 wt %. In another example, the sodium copper
chlorophyllin can be present in the formulation from about 5 wt %
to about 90 wt %. In one example, the spearmint oil can be present
in the composition from about 10 wt % to about 50 wt %. In another
example, the spearmint oil can be present in the formulation from
about 5 wt % to about 25 wt %.
[0077] The ratio of sodium copper chlorophyllin to spearmint oil
can also vary in the composition. In one example, the ratio can
range from about 1:1 to about 10:1, respectively. In one example,
the spearmint oil can be derived from the aerial parts, roots,
leaves, flowers, or a combination thereof.
[0078] In some examples, the chlorophyllin and spearmint
composition can acutely raise NO levels without administering a NOS
dependent sources of nitric oxide, such as L-arginine, citrulline
and/or without administering a NOS independent source of nitric
oxide, such as beet root extract. In some examples, the sodium
copper chlorophyllin and the spearmint oil can be administered in a
composition alone. In some examples, the composition can include
sodium copper chlorophyllin, spearmint oil, proplparben, and
methylparaben without a NOS dependent or independent source of
nitric oxide. A composition comprising sodium copper chlorophyllin,
spearmint (Mentha spicada) oil, or a combination thereof, with or
without a NOS dependent and independent source of NO, can also be
used in any of the methods described herein.
[0079] In one specific example of the composition, the NOS
dependent source of nitric oxide can include a member selected from
the group consisting of L-arginine, L-citrulline, or a combination
thereof and the myeloperoxidase inhibitor can include red grape
seed extract and pomegranate fruit extract. In yet another example,
the NOS dependent source of nitric oxide can include a member
selected from the group consisting of L-arginine, L-citrulline, or
a combination thereof the myeloperoxidase inhibitor can include red
grape seed extract and pomegranate fruit extract; and the NOS
independent source of nitric oxide can include red beet extract and
vitamin B1.
[0080] In some examples, the composition can further include
d-ribose, folic acid, malic acid, vitamin B6, vitamin B12, vitamin
D3, magnesium oxide, calcium, inulin, chicory root extract, cherry
extract, or a combination thereof. In some examples, the
composition can further include a pharmaceutically acceptable
carrier. In one example, the composition can further include
coatings, isotonic agents, absorption delaying agents, binders,
adhesives, lubricants, disintergrants, coloring agents, flavoring
agents, sweetening agents, absorbants, detergents, emulsifying
agents, antixoidants, vitamins, minerals, proteins, fats,
carbohydrates, or a combination thereof. In one example, the
composition can further include a sweetener, a preservative, a
flavoring, or a combination thereof. In some examples, the
formulation can include a polymers for sustained release of a given
compound.
[0081] In another example, the formulation can include emulsifiers.
In one example, the emulsifier can add stability to the final
product. Examples of suitable emulsifiers include, but are not
limited to, lecithin (e.g., from egg or soy), or mono- and
di-glycerides. Other emulsifiers are readily apparent to the
skilled artisan and selection of suitable emulsifier(s) will
depend, in part, upon the formulation and final product.
[0082] In yet another example, the formulation can include a
preservative. In one example, the preservatives such as potassium
sorbate, sodium sorbate, potassium benzoate, sodium benzoate, or
calcium disodium EDTA are used.
[0083] In a further example, the nutritional supplement can contain
natural or artificial sweeteners, e.g., glucose, sucrose, fructose,
saccharides, cyclamates, aspartamine, sucralose, aspartame,
acesulfame K, or sorbitol.
[0084] In one example, the composition can be in the form of an
oral dosage formulation. In another example, the oral dosage
formulation can be a capsule, a tablet, a soft gel, a lozenge, a
sachet, a powder, a beverage, a syrup, a suspension, or a food. In
another example, the compositions can be formulated into a food or
drink, and provided, for example, as a snack bar, a cereal, a
drink, a gum, or in any other easily ingested form. In one example,
the composition can be incorporated into a liquid beverage such as
water, milk, juice, or soda. In another example, the composition
can be formulated into a nutritional beverage. The nutritional
beverage can be in a premixed formulation or can be a powdered mix
in that can be added to a beverage. In another example, the powder
mix in can be in the form of granules. In one example, the
composition can be dried and made readily soluble in water.
[0085] In yet another example, the oral dosage form can be in a
solution or a suspension in an aqueous liquid or non-aqueous
liquid, such as ethanol, glycerol, vegetable oil, salt solutions,
or hydroxymethyl cellulose; or in the form of an oil-in-water
emulsion or a water-in-oil emulsion, or a combination thereof. In
examples where the oral dosage form includes oils, the oils can be
edible oils, such as e.g. cottonseed oil, sesame oil, coconut oil,
or peanut oil. In some examples, the composition can include
suitable dispersing or suspending agents for aqueous suspensions
include synthetic or natural gums such as tragacanth, alginate, gum
arabic, dextran, sodium carboxymethylcellulose, gelatin,
methylcellulose, and polyvinylpyrrolidone.
[0086] In another example, the composition can be formulated into a
food product. In one example, the food product can be a pudding,
confections, (i.e., candy), ice cream, frozen confections and
novelties, or non-baked extruded food products such as bars. In one
example, the composition can be a powder that is added to non-baked
goods. For, example a nutritional bar can be manufactured by adding
the powder to the dry ingredients and then incorporating the dry
and wet. The wet and dry ingredients can be mixed until the dough
phase is reached. The dough can then be put into an extruder and
extruded; the extruded dough can be cut into appropriate lengths;
and the product can be cooled.
[0087] Flavors, coloring agents, spices, nuts, and the like can be
incorporated into the product. Flavorings can be in the form of
flavored extracts, volatile oils, chocolate flavorings (e.g.,
non-caffeinated cocoa or chocolate, chocolate substitutes such as
carob), peanut butter flavoring, cookie crumbs, crisp rice,
vanilla, or any commercially available flavoring. Flavorings can be
protected with mixed tocopherols. Examples of useful flavorings
include but are not limited to pure anise extract, imitation banana
extract, imitation cherry extract, chocolate extract, pure lemon
extract, pure orange extract, pure peppermint extract, imitation
pineapple extract, imitation rum extract, imitation strawberry
extract, or pure vanilla extract; or volatile oils, such as balm
oil, bay oil, bergamot oil, cedarwood oil, cherry oil, walnut oil,
cinnamon oil, clove oil, or peppermint oil; peanut butter,
chocolate flavoring, vanilla cookie crumb, butterscotch, or toffee.
In one example, the nutritional supplement contains berry or other
fruit flavor. The food compositions may further be coated, for
example with a yogurt coating if it is as a bar.
[0088] In one example, the oral dosage form can include from about
5 mg to about 1,000 mg of the NOS dependent source of nitric oxide,
from about 5 mg to about 1,000 mg of the NOS independent source of
nitric oxide, and from about 5 mg to about 100 mg of the
myeloperoxidase inhibitor. In another example, the oral dosage form
can include from about 1 mg to about 100 mg of the NOS dependent
source of nitric oxide, from about 1 mg to about 100 mg of the NOS
independent source of nitric oxide, and from about 1 mg to about
100 mg of the myeloperoxidase inhibitor. In yet another example,
the oral dosage form can include from about 0.5 mg to about 100 mg
of the NOS dependent source of nitric oxide, from about 0.5 mg to
about 100 mg of the NOS independent source of nitric oxide, and
from about 0.5 mg to about 100 mg of the myeloperoxidase inhibitor.
In yet another example, the oral dosage form is prepared for
administration to the subject according to a predetermined regimen.
In a further example, the oral dosage form can be formulated to be
administered to the subject once per day.
[0089] In one example, the formulation can be in the form of a
cream or lotion for topical application. In another example the
active ingredient can be in the form of a bolus, electuary, or
paste. In yet another example, the composition can be formulated as
a depot preparation. In one example, the depot can be for
implantation (e.g. subcutaneously, intra-abdominally, or
intramuscularly) or intramuscular injection. In one example, the
formulation can be formulated as an ion exchange resin.
[0090] In one example, the composition can provide an acute
increase in biosynthesis of nitric oxide and/or inhibit the
activity of myeloperoxidase. In another example, the composition
can acutely raise nitric oxide levels in the subject by enhancing
biosynthesis production of nitric oxide. In yet another example,
the composition can acutely raise nitric oxide levels in the
subject by inhibiting myeloperoxidase activity. In a further
example, the composition can acutely raise nitric oxide levels in
the subject by providing a nitrite/nitrate source for conversion to
nitric oxide.
[0091] Further presented herein, is a method for acutely raising
nitric oxide levels in a subject. In one example, the method can
include administering to the subject a therapeutically effective
amount of any of the compositions previously presented or within
the examples section.
[0092] In another example, a method of treating a subject for a
condition or disorder that is response to nitric oxide therapy is
presented. In one example, the method can include: acutely raising
nitric oxide levels in a subject by simultaneously increasing
biosynthesis of nitric oxide, increasing nitrate/nitrite levels,
and inhibiting myeloperoxidase activity. In one example, the method
can include administering a therapeutically effective amount of the
composition above, the NOS dependent source of nitric oxide, the
NOS independent source of nitric oxide, the myeloperoxidase
inhibitor, or a combination thereof.
[0093] In one example, the condition or disorder is a nitric oxide
related pathology. In another example, the nitric oxide related
pathology can include: Alzheimer's disease, angina, asthma,
congestive disorders, Crohn's disease, deep vein thrombosis,
dementia, diabetes (types, 1, 2 and 3), diabetic foot disorders,
diminished exercise capacity, endothelial dysfunction, endotoxemia,
erectile dysfunction, fibromyalgia, heart attack, heart failure,
hypertension, inflammatory bowel disease, leaky gut, macular
degeneration, monocyte-mediated arterial plaque formation, motor
dysfunction, multiple sclerosis, obesity, oxidation of LDL,
periodontal disease, peripheral arterial disease, platelet
stickiness, portal hypertension, pregnancy/pre-eclampsia, premature
ejaculation, pulmonary hypertension, Raynaud's disease, renal
failure, sleep apnea, smooth muscle cell proliferation, stroke, and
vasculitis.
[0094] In a further example, the condition or disorder can be a
cardio-metabolic disorder. In one example, the cardio-metabolic
disorder can include: Alzheimer's disease, angina, asthma,
congestive disorders, Crohn's disease, deep vein thrombosis,
dementia, diabetes (types, 1, 2 and 3), diabetic foot disorders,
diminished exercise capacity, endothelial dysfunction, endotoxemia,
erectile dysfunction, fibromyalgia, heart attack, heart failure,
hypertension, inflammatory bowel disease, leaky gut, macular
degeneration, monocyte-mediated arterial plaque formation, motor
dysfunction, multiple sclerosis, obesity, oxidation of LDL,
periodontal disease, peripheral arterial disease, platelet
stickiness, portal hypertension, pregnancy/pre-eclampsia, premature
ejaculation, pulmonary hypertension, Raynaud's disease, renal
failure, sleep apnea, smooth muscle cell proliferation, stroke, and
vasculitis. In yet another example, the cardio-metabolic disorder
can include hypertension, cardiovascular dysfunction,
neurodegeneration, arthritis, asthma, and septic shock. In a
further example, the cardio-metabolic disorder can be preventing
the formation of arterial plaque. In one example, the treatment of
the cardio-metabolic disorder can be prophylactic.
[0095] In one example, the condition or disorder can be a
myeloperoxidase related pathology. In another example, the
myeloperoxidase-related pathology can be Alzheimer's disease,
angina, asthma, general congestive disorders, Crohn's disease, deep
vein thrombosis, dementia, diabetes (types, 1, 2 and 3), diabetic
foot disorders, diminished exercise capacity, endothelial
dysfunction, endotoxemia, erectile dysfunction, fibromyalgia
glomerulonephritis, heart attack, heart failure, hypertension,
immune deficiency, inflammatory bowel disease, leaky gut, macular
degeneration, monocyte-mediated arterial plaque formation, motor
dysfunction, multiple sclerosis, obesity, oxidation of LDL,
periodontal disease, peripheral arterial disease, platelet
stickiness, portal hypertension, pregnancy/pre-eclampsia, premature
ejaculation, pulmonary hypertension, Raynaud's disease, renal
failure, sleep apnea, smooth muscle cell proliferation, stroke,
vasculitis and diseases associated with skin such as slow wound
healing, wrinkles, and premature signs of aging. In a further
example, the myeloperoxidase related pathology can be increased
oxidized LDL cholesterol. In yet a further example, the
myeloperoxidase related pathology can be metabolic syndrome, type 1
diabetes, type 2 diabetes, type 3 diabetes, or a combination
thereof. In one example, the myeloperoxidase related pathology can
be leaky gut, endotoxemia, inflammatory bowel disease, or a
combination thereof. In a further example, the myeloperoxidase
related pathology can be a dermatopic pathology including slow
wound healing, wrinkles, sun spots, and premature signs of
aging.
[0096] In some examples, the treatment of the subject can be
prophylactic. In one example, the condition or disorder can be
penile dysfunction. In some examples, acutely raising of nitric
oxide levels in the subject can enhance endothelial functioning,
decrease monocyte-mediated arterial plaque formation, decrease the
development of peripheral arterial disease, or a combination
thereof, wherein an increase or decrease refers to a level in the
subject prior to the administering of the therapeutically effective
combination.
[0097] In one example, the method can be used to treat a mammal. In
one example, the mammal can be a human. In another example, the
method can be used to treat a domestic animal, such as a dog or
cat. In yet another example, the method can be used to treat
livestock, such as cows, horses, donkeys, or pigs. The treatment
can in some examples be prophylactic. In some examples, acutely
raising nitric oxide levels in the subject can raise salivary
nitrite levels in the subject beyond a level of the salivary
nitrite in the subject as compared to a level prior to
administering the therapeutically effective combination.
[0098] Further presented herein is a system for acutely raising
nitric oxide levels in a subject. In one example, the system can
include: an effective amount of a NOS dependent source of nitric
oxide, an effective amount of a NOS independent source of nitric
oxide, and an effective amount of a myeloperoxidase inhibitor. In
one example, the at least one of the NOS dependent source of nitric
oxide, the NOS independent source of nitric oxide, and the
myeloperoxidase inhibitor can be separate from one another. In
another example, the at least one of the NOS dependent source of
nitric oxide, the NOS independent source of nitric oxide, and the
myeloperoxidase inhibitor can be in separate formulations.
[0099] In some examples, the level of nitric oxide in the subject
following administration of the composition in the system can be
greater than an amount provided by an equivalent amount of any one
of the NOS dependent source of nitric oxide, the NOS independent
source of nitric oxide, or the myeloperoxidase inhibitor. In
another example, the level of nitric oxide can be greater than an
additive amount of an equivalent amount of any one of the NOS
dependent source of nitric oxide, the NOS independent source of
nitric oxide, or the myeloperoxidase inhibitor.
[0100] In yet a further example, the system can be formulated as
kit. In one example, the kit can include one or more containers
filled with one or more of the ingredients of the compositions.
Optionally associated with such container(s), can be a notice in
the form prescribed by a government agency regulating the
manufacture, use or sale of pharmaceutical products, which notice
reflects approval by the agency of manufacture, use of sale, and/or
approval for human administration. The pack or kit can be labeled
with information and instructions regarding mode of administration,
sequence of administration (e.g., separately, sequentially, or
concurrently), dosing regimen, or the like. The pack or kit may
also include means for reminding the patient to take the therapy.
The pack or kit can be a single unit dosage of the combination
therapy or it can be a plurality of unit dosages. In particular,
the agents can be separated, mixed together in any combination,
present in a formulation or tablet.
[0101] The recommended daily amounts of each ingredient, can serve
as a guideline for formulating the compositions and systems of this
invention. The actual amount of each ingredient per unit dosage
will depend upon the number of units administered daily to the
individual in need thereof. This is a matter of product design and
is well within the skill of the nutritional supplement
formulator.
[0102] The phytocomplexes described herein have few obvious
physiochemical properties in common. Functionally, however, all
have demonstrated a combination of chemical and biological
characteristics necessary for attenuating the role of MPO in eNOS
biosynthesis of NO. As depicted in FIG. 8A, MPO can functionally
uncouple eNOS resulting in the production of singlet state
O.sub.2.sup.-, as well as, NO by eNOS. The NO and O.sub.2.sup.- can
then combine to form peroxynitrite (ONOO..sup.-), which in turn
nitrates proteins and disrupts metabolic homeostasis. MPO can also
form hypochlorous acid (HOCL) as previously described, which also
functions to disrupt metabolic homeostasis through protein
nitration.
[0103] The phytocomplexes of the instant invention all appear to
simultaneously function (1) to chemically eliminate ONOO..sup.- (X1
site in FIG. 8B), (2) to enzymatically inhibit MPO formation of
hypochlorite (X2 site in FIG. 8B), and (3) to enzymatically inhibit
MPO uncoupling of eNOS (X3 site in FIG. 8B). This novel,
multi-targeted effect of the phytocomplexes described herein offers
a cogent explanation for the ability of formulations that do not
contain dietary NO.sub.3 or the amino acids arginine and citrulline
to rapidly increase NO. Inhibiting the uncoupling of eNOS by MPO
would prevent the loss of NO through the O.sub.2.sup.- and
ONOO..sup.- pathway. Similarly, the phytocomplexes would be
expected to enhance the effects of either dietary NO.sub.3 or
arginine and citrulline, as was demonstrated herein in clinical
studies. Thus, a unique multi-targeted property of specific
phytocomplexes has been discovered with uses in the rapid
enhancement of NO biosynthesis or bioavailability.
EMBODIMENTS
[0104] In one embodiment presented herein, is a composition for
acutely raising nitric oxide levels in a subject, comprising: an
effective amount of a NOS dependent source of nitric oxide; an
effective amount of a NOS independent source of nitric oxide; and
an effective amount of a myeloperoxidase inhibitor; wherein the
composition acutely raises nitric oxide levels in a subject above a
level provided by the available sources of nitric oxide in the
subject prior to administration of the composition.
[0105] In one embodiment of the composition, the level of nitric
oxide in the subject following administration of the composition is
greater than an amount provided by an equivalent amount of any one
of the NOS dependent source of nitric oxide, the NOS independent
source of nitric oxide, or the myeloperoxidase inhibitor.
[0106] In one embodiment of the composition, the NOS dependent
source of nitric oxide comprises from about 1 wt % to about 80 wt %
of the composition.
[0107] In one embodiment of the composition, the NOS dependent
source of nitric oxide comprises a member selected from the group
consisting of L-arginine, L-citrulline, ornithine, or a combination
thereof.
[0108] In one embodiment of the composition, the NOS dependent
source of nitric oxide comprises L-arginine.
[0109] In one embodiment of the composition, the L-arginine in the
composition ranges from about 1 wt % to about 80 wt %.
[0110] In one embodiment of the composition, the NOS dependent
source of nitric oxide comprises L-citrulline.
[0111] In one embodiment of the composition, the L-citrulline in
the composition ranges from about 1 wt % to about 80 wt %.
[0112] In one embodiment of the composition, a source of the
L-citrulline comprises watermelon extract.
[0113] In one embodiment of the composition, the NOS dependent
source of nitric oxide comprises L-arginine and L-citrulline.
[0114] In one embodiment of the composition, the L-arginine and the
L-citrulline each comprise from 1 wt % to about 80 wt %.
[0115] In one embodiment of the composition, the L-arginine and
L-citrulline are present in the composition at a weight percent
concentration range from about 1:5 to about 5:1, respectively.
[0116] In one embodiment of the composition, the L-arginine and
L-citrulline are present in the composition at a weight percent
concentration ratio of about 2:1, respectively.
[0117] In one embodiment of the composition, the NOS independent
source of nitric oxide comprises a member selected from the group
consisting of beet root extract, vitamin B1, collard green extract,
nut powders, spinach extract, broccoli extract, lettuce extract,
celery, kale, watercress, carrot, arugula, mustard greens, or a
combination thereof.
[0118] In one embodiment of the composition, the NOS independent
source of nitric oxide comprises beet root extract, vitamin B1, or
a combination thereof.
[0119] In one embodiment of the composition, the NOS independent
source of nitric oxide comprises the beet root extract and the
vitamin B1.
[0120] In one embodiment of the composition, the beet root extract
comprises from about 5 wt % to about 90 wt % and wherein the
vitamin B1 comprises from about 0.01 wt % to about 80 wt % of the
composition.
[0121] In one embodiment of the composition, the beet root extract
and vitamin B1 are present in the composition at a weight percent
concentration ratio ranging from about 50:1 to about 10:1,
respectively.
[0122] In one embodiment of the composition, the myeloperoxidase
inhibitor comprises from about 1 wt % to about 90 wt %.
[0123] In one embodiment of the composition, the myeloperoxidase
inhibitor comprises a member selected from the group consisting of
pomegranate fruit extract, red grape polyphenols, apple extract,
blueberry extract, capsicum extract, grape extract, green tea
extract, olive extract, bergamot extract, mangosteen, or a
combination thereof.
[0124] In one embodiment of the composition, the mangosteen can
comprise a member selected from the group consisting of mangosteen
fruit, mangosteen extract, or a combination thereof.
[0125] In one embodiment of the composition, the myeloperoxidase
inhibitor comprises a member selected from the group consisting of
pomegranate fruit extract, red grape polyphenols, or a combination
thereof.
[0126] In one embodiment of the composition, the myeloperoxidase
inhibitor comprises pomegranate fruit extract, and red grape
polyphenols.
[0127] In one embodiment of the composition, the pomegranate fruit
extract comprises from about 1 wt % to about 80 wt % and wherein
the red grape polyphenols comprises from about 1 wt % to about 80
wt % of the composition.
[0128] In one embodiment of the composition, the myeloperoxidase
inhibitor comprises apple extract, grape extract, green tea
extract, and olive extract.
[0129] In one embodiment of the composition, the apple extract,
grape extract, green tea extract, and olive extract collectively
comprise from about 1 wt % to about 80 wt % of the composition.
[0130] In one embodiment of the composition, the apple extract
comprises from about 0.01 wt % to about 80 wt %, the grape extract
comprises from about 0.01 wt % to about 80 wt %, the green tea
extract comprises from about 0.01 wt % to about 80 wt % and the
olive extract comprises from about 0.01 wt % to about 80 wt % of
the claimed composition.
[0131] In one embodiment of the composition, the apple extract,
grape extract, green tea extract, and olive extract are present in
the composition at a weight ratio of about 1:1:1:1.
[0132] In one embodiment of the composition, the apple extract,
grape extract, green tea extract, and olive extract are present in
the composition at a weight ratio of about 6:1:3:1.
[0133] In one embodiment of the composition, at least one of the
apple extract, the grape extract, the green tea extract, and the
olive extract are present in the composition in a different
amount.
[0134] In one embodiment of the composition, the grape extract
comprises grape seed extract and grape skin extract.
[0135] In one embodiment of the composition, the myeloperoxidase
inhibitor further comprises pomegranate fruit extract.
[0136] In one embodiment of the composition, the myeloperoxidase
inhibitor further comprises blueberry fruit extract, capsicum fruit
extract, and turmeric root extract.
[0137] In one embodiment of the composition, the blueberry fruit
extract comprises from about 0.01 wt % to about 80 wt %, the
capsicum fruit extract comprises from about 0.01 wt % to about 80
wt %, and the turmeric root extract comprises from about 0.01 wt %
to about 80 wt % of the composition.
[0138] In one embodiment of the composition, the apple extract, the
grape extract, the green tea extract, the olive extract, the
blueberry fruit extract, the capsicum fruit extract, and the
turmeric root extract are present in the composition at a weight
ratio of about 1:1:1:1:1:1:1.
[0139] In one embodiment of the composition, the composition can
further comprises turmeric rhizome extract, and mangosteen
extract.
[0140] In one embodiment of the composition, the turmeric rhizome
extract comprises from about 0.1 wt % to about 80 wt % and the
mangosteen extract comprises from about 0.1 wt % to about 80 wt %
of the composition.
[0141] In one embodiment of the composition, the apple extract, the
grape extract, the green tea extract, the olive extract, the
blueberry fruit extract, the capsicum fruit extract, the turmeric
root extract, the turmeric rhizome extract, and the mangosteen are
present in the composition at a weight ratio of about
1:1:1:1:1:1:1:1:1.
[0142] In one embodiment of the composition, the mangosteen
comprises a mangosteen fruit, a mangosteen pericarp, a manogsteen
extract, or a combination thereof.
[0143] In one embodiment of the composition, the formulation
further comprises bergamot fruit extract.
[0144] In one embodiment of the composition, the bergamot fruit
extract comprises from about 0.01 wt % to about 80 wt % of the
composition.
[0145] In one embodiment of the composition, the myeloperoxidase
inhibitor can comprise sodium copper chlorophyllin, spearmint oil,
or a combination thereof.
[0146] In one embodiment of the composition, the NOS dependent
source of nitric oxide comprises a member selected from the group
consisting of L-arginine, L-citrulline, or a combination thereof
and wherein the myeloperoxidase inhibitor comprises red grape seed
extract and pomegranate fruit extract.
[0147] In one embodiment of the composition, the composition
further comprises a member selected from the group consisting of
d-ribose, folic acid, malic acid, vitamin B6, vitamin B12, vitamin
D3, magnesium oxide, calcium, inulin, chicory root extract, cherry
extract, or a combination thereof.
[0148] In one embodiment of the composition, the composition
further comprises a pharmaceutically acceptable carrier.
[0149] In one embodiment of the composition, the composition
further comprises a member selected from the group consisting of a
sweetener, a preservative, a flavoring, or a combination
thereof.
[0150] In one embodiment of the composition, the composition is an
oral dosage formulation
[0151] In one embodiment of the composition, the oral dosage form
comprises a capsule, a tablet, a powder, a beverage, a syrup, a
suspension, or a food.
[0152] In one embodiment of the composition, the oral dosage form
comprises from about 5 mg to about 1,000 mg of the NOS dependent
source of nitric oxide, from about 5 mg to about 1,000 mg of the
NOS independent source of nitric oxide, and from about 5 mg to
about 1,000 mg of the myeloperoxidase inhibitor.
[0153] In one embodiment of the composition, the oral dosage form
is prepared for administration to the subject according to a
predetermined regimen.
[0154] In one embodiment of the composition, the oral dosage form
is administered to the subject once per day.
[0155] In one embodiment of the composition, the composition
acutely raises nitric oxide levels in the subject by enhancing
biosynthesis production of nitric oxide.
[0156] In one embodiment of the composition, composition acutely
raises nitric oxide levels in the subject by inhibiting
myeloperoxidase activity.
[0157] In one embodiment of the composition, the composition
acutely raises nitric oxide levels in the subject by providing a
nitrite/nitrate source for conversion to nitric oxide.
[0158] In another embodiment presented herein is a method for
acutely raising nitric oxide levels in a subject, comprising:
administering to the subject a therapeutically effective amount of
any one of the compositions recited above.
[0159] In yet another embodiment presented herein, is a method of
treating a subject for a condition or disorder that is response to
nitric oxide therapy, comprising: acutely raising nitric oxide
levels in a subject by simultaneously increasing biosynthesis of
nitric oxide, increasing nitrate/nitrite levels, and inhibiting
myeloperoxidase activity.
[0160] In one embodiment of the method, the condition or disorder
is a nitric oxide related pathology.
[0161] In one embodiment of the method, the nitric oxide related
pathology comprises a member selected from the group consisting of
Alzheimer's disease, angina, asthma, congestive disorders, Crohn's
disease, deep vein thrombosis, dementia, diabetes (types, 1, 2 and
3), diabetic foot disorders, diminished exercise capacity,
endothelial dysfunction, endotoxemia, erectile dysfunction,
fibromyalgia, heart attack, heart failure, hypertension,
inflammatory bowel disease, leaky gut, macular degeneration,
monocyte-mediated arterial plaque formation, motor dysfunction,
multiple sclerosis, obesity, oxidation of LDL, peridontal disease,
peripheral arterial disease, platelet stickiness, portal
hypertension, pregnancy/pre-eclampsia, premature ejaculation,
pulmonary hypertension, Raynaud's disease, renal failure, sleep
apnea, smooth muscle cell proliferation, stroke, and
vasculitis.
[0162] In one embodiment of the method, the condition or disorder
is a cardio-metabolic disorder.
[0163] In one embodiment of the method, the cardio-metabolic
disorder comprises a member selected from the group consisting of:
Alzheimer's disease, angina, asthma, congestive disorders, Crohn's
disease, deep vein thrombosis, dementia, diabetes (types, 1, 2 and
3), diabetic foot disorders, diminished exercise capacity,
endothelial dysfunction, endotoxemia, erectile dysfunction,
fibromyalgia, heart attack, heart failure, hypertension,
inflammatory bowel disease, leaky gut, macular degeneration,
monocyte-mediated arterial plaque formation, motor dysfunction,
multiple sclerosis, obesity, oxidation of LDL, peridontal disease,
peripheral arterial disease, platelet stickiness, portal
hypertension, pregnancy/pre-eclampsia, premature ejaculation,
pulmonary hypertension, Raynaud's disease, renal failure, sleep
apnea, smooth muscle cell proliferation, stroke, and
vasculitis.
[0164] In one embodiment of the method, the cardio-metabolic
disorder comprises a member selected from the group consisting of
hypertension, cardiovascular dysfunction, neurodegeneration,
arthritis, asthma, and septic shock.
[0165] In one embodiment of the method, the cardio-metabolic
disorder comprises preventing the formation of arterial plaque.
[0166] In one embodiment of the method, the treating of the subject
is prophylactic.
[0167] In one embodiment of the method, the condition or disorder
is a myeloperoxidase related pathology.
[0168] In one embodiment of the method, the myeloperoxidase-related
pathology is Alzheimer's disease, angina, asthma, general
congestive disorders, Crohn's disease, deep vein thrombosis,
dementia, diabetes (types, 1, 2 and 3), diabetic foot disorders,
diminished exercise capacity, endothelial dysfunction, endotoxemia,
erectile dysfunction, fibromyalgia glomerulonephritis, heart
attack, heart failure, hypertension, immune deficiency,
inflammatory bowel disease, leaky gut, macular degeneration,
monocyte-mediated arterial plaque formation, motor dysfunction,
multiple sclerosis, obesity, oxidation of LDL, peridontal disease,
peripheral arterial disease, platelet stickiness, portal
hypertension, pregnancy/pre-eclampsia, premature ejaculation,
pulmonary hypertension, Raynaud's disease, renal failure, sleep
apnea, smooth muscle cell proliferation, stroke, vasculitis and
diseases associated with skin such as slow wound healing, wrinkles,
and premature signs of aging.
[0169] In one embodiment of the method, the myeloperoxidase related
pathology comprises increased oxidized LDL cholesterol.
[0170] In one embodiment of the method, the myeloperoxidase related
pathology comprises metabolic syndrome, type 1 diabetes, type 2
diabetes, type 3 diabetes, or a combination thereof.
[0171] In one embodiment of the method, the myeloperoxidase related
pathology comprises leaky gut, endotoxemia, inflammatory bowel
disease or a combination thereof.
[0172] In one embodiment of the method, the myeloperoxidase related
pathology comprises a dermatopic pathology including slow wound
healing, wrinkles, sun spots, and premature signs of aging.
[0173] In one embodiment of the method, the treating of the subject
is prophylactic.
[0174] In one embodiment of the method, the condition or disorder
is penile dysfunction.
[0175] In one embodiment of the method, acutely raising of nitric
oxide levels in the subject enhances endothelial functioning,
decreases monocyte-mediated arterial plaque formation, decreases
the development of peripheral arterial disease, or a combination
thereof, wherein an increase or decreases refers to a level in the
subject prior to the administering of the therapeutically effective
combination.
[0176] In one embodiment of the method, the subject is a human.
[0177] In one embodiment of the method, the treating of the subject
is prophylactic.
[0178] In one embodiment of the method, acutely raising nitric
oxide levels in the subject comprises raising salivary nitrite
levels in the subject beyond a level of the salivary nitrite in the
subject as compared to a level prior to administering the
therapeutically effective combination.
[0179] In another embodiment presented herein is a system for
acutely raising nitric oxide levels in a subject, comprising: an
effective amount of a NOS dependent source of nitric oxide; an
effective amount of a NOS independent source of nitric oxide; and
an effective amount of a myeloperoxidase inhibitor.
[0180] In one embodiment of the system, at least one of the NOS
dependent source of nitric oxide, the NOS independent source of
nitric oxide, and the myeloperoxidase inhibitor are separate from
one another.
[0181] In one embodiment of the system, at least one of the NOS
dependent source of nitric oxide, the NOS independent source of
nitric oxide, and the myeloperoxidase inhibitor are in separate
formulations.
[0182] In one embodiment of the system, a level of nitric oxide in
the subject following administration of the composition is greater
than an amount provided by an equivalent amount of any one of the
NOS dependent source of nitric oxide, the NOS independent source of
nitric oxide, or the myeloperoxidase inhibitor.
[0183] In one embodiment of the system, the level of nitric oxide
is greater than an additive amount of an equivalent amount of any
one of the NOS dependent source of nitric oxide, the NOS
independent source of nitric oxide, or the myeloperoxidase
inhibitor.
[0184] In one embodiment of the system, the system is formulated as
a kit.
[0185] In one embodiment presented herein is, a composition for
acutely raising nitric oxide levels in a subject, comprising an
effective amount of sodium copper chlorophyllin, spearmint oil, or
a combination thereof.
[0186] In one embodiment, the composition comprises sodium copper
chlorophyllin and spearmint oil.
[0187] In one embodiment, the composition comprises a weight ratio
of sodium copper chlorophyllin to spearmint oil ranges from about
1:1 to about 10:1.
[0188] In one embodiment, the composition comprises spearmint oil
and the spearmint oil is derived from the aerial parts, roots,
leaves, flowers, or a combination thereof.
[0189] In one embodiment, a method of treating a subject for a
condition or disorder that is response to nitric oxide therapy,
comprising inhibiting meyloperoxidase activity is provided.
[0190] In one embodiment, the method can comprise administering a
therapeutically effective amount of sodium copper chlorophyllin,
spearmint oil, or a combination thereof to the subject.
EXAMPLES
Example 1
Acute Nitric Oxide Production by an
Arginine/Citrulline/Phytochemical Formulation
[0191] Two NO-generating formulations, shown below, were tested to
assess their acute, enhanced biosynthesis of the NO biomarker,
NO.sub.2 in saliva in subjects with normal plasma concentrations of
ADMA.
TABLE-US-00002 TABLE 2 Active Ingredients in Test Formulations
Formula 1 Formula 2 [Active [Active Active Ingredient [mg]
Fraction] [mg] Fraction] L-Arginine 5,300 0.92 5,008 0.87
L-Citrulline 250 0.043 209 0.036 Pomegranate Juice 50 0.0087 491
0.085 Concentrate Grape Skin Extract 185 0.0321 73.6 0.0126 Red
Grape Polyphenol 0.00 0.00 49.1 0.87
Methods--The Nitrate/Nitrite Fluorometric Assay Kit (Cayman
Chemicals Item No: 780051, Ann Arbor, Mich.) was used to measure
the NO.sub.3/NO.sub.2 content of the formulations. Fluorescence was
analyzed with an excitation wavelength of 375 nm and an emission
wavelength of 417 nm using a Cytation5 microplate fluorometer (Bio
Tek Instruments, Winooski, Vt.). The assay has a reported limit of
detection for NO.sub.3 of approximately 0.2 .mu.M in the final
solution. Over three days, subjects, with normal plasma
concentrations of ADMA, were instructed to consume the 10 g of
either F1 or F2 with 500 mL of water subsequent to developing the
first, pre-dose NO strip. At post-dosing times 30, 60 and 90
minutes, NO strips were again developed for all subjects. One
subject was given a 2.times. dose (20 g) of both formulations over
the experimental period. The salivary NO strips, which detect
salivary NO.sub.2, were nitric Oxide Test Strips (Berkeley Test,
Berkeley, Calif.). Color development of NO strips was quantitated
using densitometry. Means and 95% confidence intervals were
computed using Excel [Microsoft, Redman, Wash.]. Results--As seen
in FIG. 9A, F2 unexpectedly produced an average increase of 21%
relative to F1 at 30 min (p<0.5). The acute increase in NO
biomarker with F2 continued with a 35% increase relative to zero
time F2 and 66% relative to F1 at 60 min (p<0.05). While F2 NO
biomarker levels remained 19% above F2 zero time at 90 min,
NO.sub.2 levels were 49% above F1 concentrations (p<0.05) at
this final time point.
[0192] NO biomarker results for the subject consuming the
double-dose of both F1 and F2 are presented in FIG. 9B. As seen
with single-doses, F2 produced an acute NO strip response of 73%
within 30 min that continued increasing through 60 min to 2.5-times
the pre-dose value. Unlike the single-dose, however, the NO
biomarker concentration did not fall at 90 min, but remained at the
60-min level. Interestingly, the double dose of F1 resulted in a
42% increase in NO biomarker at 90 min post-dosing, but remained at
pre-dose levels at 30 and 60 min post-dosing.
[0193] Nitrate levels for F1 and F1 were below the limit of
detection of the assay. Samples of pomegranate juice concentration
and red grape polyphenol contained, respectively, 24 and 70 .mu.g
NO.sub.3/g. The addition of pomegranate juice concentration and red
grape polyphenol to F2 only increased the NO.sub.3 concentration to
approximately 1 mg/10 g dosing. Based upon current pharmacodynamic
studies in this laboratory, sample NO.sub.3 content below 12.5 mg
NO.sub.3/dose do not produce a detectable increase in NO strip
color development. Therefore, it is unlikely that the level of
NO.sub.3 in F2 could be responsible for the acute increase in NO
biomarker observed for F2.
[0194] A unique blend of arginine, citrulline and antioxidants
produced an acute increase in the salivary NO biomarker NO.sub.2,
indicative of a rapid biosynthesis of NO.
Example 2
Synergistic Inhibition of Myeloperoxidase Activity by an
Arginine/Citrulline Formulation
[0195] Formulations 1 and 2, as described in Table 2 above, were
tested for their synergistic activity. Additional samples of F1 and
F2 were prepared at an initial concentration of 100 mg/mL in an
acetone:water (1:1) solution, sonicated in an ice bath for 60 min
with vortexing every 10 min, and centrifuged at 3000 rpm at ambient
temperature for 5 min. The supernatant liquid was transferred to
fresh tubes and centrifuged again at 13,300 rpm at ambient
temperature for 10 min. This final supernatant fraction was
transferred to fresh tubes and stored at -80.degree. C. until
assayed.
[0196] The acetone used in this example was obtained through Fisher
Chemical Co (Pittsburgh, Pa.) and was the highest purity available.
All other chemicals and reagents were supplied with the
Myeloperoxidase Inhibitor Screening Assay Kit. The Myeloperoxidase
Control was stored at -20.degree. C., while the remainder of the
kit was stored at -4.degree. C. The positive control
4-aminobenzyhdrazide, supplied with the kit, was run with each
assay.
Methodology--The Myeloperoxidase Inhibitor Screening Assay Kit Item
No. 700170 from Cayman Chemical (Ann Arbor, Mich.) was used to
assess the ability of the two arginine/citrulline formulations and
their components individually to inhibit the production of
hypochlorous acid by MPO, as can be seen in FIG. 9 (A&B).
[0197] The chlorination assay utilizes the non-fluorescent
2-[6-(4-aminophenoxy)-3-oxo-3H-xanthen-9-yl]-benzoic acid (APF),
which is selectively cleaved by hypochlorite to yield the highly
fluorescent compound fluorescein. Fluorescein fluorescence was
analyzed with an excitation wavelength of 480-495 nm and an
emission wavelength of 515-525 nm using a Cytation5 microplate
fluorescence reader (Bio Tek Instruments, Winooski, Vt.).
Calculations--The median inhibitory concentration (IC50) for the
inhibition of MPO activity was calculated using CalcuSyn (BIOSOFT,
Ferguson, Mo.). This statistical package performs multiple drug
dose-effect calculations using the median effect methods described
by T-C Chou and P. Talaly [Chou, T-C, Talaly, P. (1984)
Quantitative analysis of dose-effect relationships: the combined
effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul
22, 27-55].
[0198] Briefly, the analysis correlates the "Dose" and the "Effect"
in the simplest possible form: fa/fu=(C/Cm)m, where C is the
concentration or dose of the compound and Cm is the
median-effective dose signifying the potency. Cm is determined from
the x-intercept of the median-effect plot. The fraction affected by
the concentration of the test material is fa and the fraction
unaffected by the concentration is fu (fu=1-fa). The exponent m is
the parameter signifying the sigmoidicity or shape of the
dose-effect curve; it is estimated by the slope of the
median-effect plot.
[0199] The median-effect plot is a graph of x=log(C) vs
y=log(fa/fu) and is based on the logarithmic form of Chou's
median-effect equation. The goodness of fit for the data to the
median-effect equation is represented by the linear correlation
coefficient r of the median-effect plot. Usually, the experimental
data from enzyme or receptor systems have an r>0.96, from tissue
culture an r>0.90 and from animal systems r>0.85.
[0200] Synergy of test components was then quantified using the
combination index (CI) parameter. This parameter defines only the
additive effect rather than synergism or antagonism. Synergy,
however, was defined as a more than expected additive effect
(CI>1), and antagonism as a less than expected additive effect
(CI<1) as described below.
[0201] Expected median inhibitory concentrations of any
multi-component combination were estimated using the
relationship:
[1/Expected IC50]=[Fa/IC50A]+[Fb/IC50B]+ . . . +[Fn/IC50N]
Fa=mole fraction of component A in the combination and Fn=the mole
fraction of the n.sup.th component combination where Fa+Fb+ . . .
+Fn=land where IC50A=the observed IC50 of the component A, etc. The
CI was then calculated thusly, CI=Expected [IC50]/Observed [IC50].
Using the designation of CI=1 as the additive effect, we obtain for
mutually exclusive compounds that have the same mode of action or
for mutually non-exclusive drugs that have totally independent
modes of action the following relationships: CI<1, =1, and >1
indicating antagonism, additivity and synergy, respectively.
Results--The observed median inhibitory concentration (IC.sub.50)
of F1 was 9.31 .mu.g/mL, while the calculated, expected IC.sub.50
was 8.57 .mu.g/mL resulting in a CI=0.92. Thus, the Formulation F1
was no better inhibiting MPO than the sum of its active components
(Table 3). On the other hand, Formula 2 inhibited MPO more
effectively than the sum of its components with an observed IC50 of
3.57 .mu.g/mL versus an expected IC50 of 8.92 and a CI=2.50 (Table
4).
TABLE-US-00003 TABLE 3 Combination Index of Formulation 1 for
Myeloperoxidase Inhibition Observed IC.sub.50.dagger. Fraction of
Test Material [.mu.g/mL] Formulation 1 L-Arginine 8.09 (6.20-10.6)
0.909 L-Citrulline 18.0 (13.7-23.6) 0.043 Pomegranate Juice 135
(92.3-197) 0.0086 Concentrate Grape Skin 34.5 (1.17-1.54) 0.0317
Extract Red Grape 1.35 (1.18-1.55) 0.00 Polyphenol Combination
Index Expected [Expected IC.sub.50 IC.sub.50/ [.mu.g/mL] Observed
IC.sub.50] Formula F1 9.31* (8.05-10.8) 8.57 0.921
.dagger.Parenthetic values are 95% confidence interval. *Observed
IC50 of the Formulation1 is not different from the Expected IC50
with a CI = 0.921 indicating a lack of synergy.
TABLE-US-00004 TABLE 4 Combination Index of Formulation 2 for
Myeloperoxidase Inhibition Observed IC.sub.50.dagger. Fraction of
Test Material [.mu.g/mL] Formulation 2 L-Arginine 8.09 (6.20-10.6)
0.859 L-Citrulline 18.0 (13.7-23.6) 0.036 Pomegranate Juice 135
(92.3-197) 0.084 Concentrate Grape Skin Extract 34.5 (1.17-1.54)
0.0126 Red Grape 1.35 (1.18-1.55) 0.0087 Polyphenol Combination
Index Expected [Expected IC.sub.50 IC.sub.50/ [.mu.g/mL] Observed
IC.sub.50] Formula F2 3.57* (2.63-4.84) 8.92 2.50
.dagger.Parenthetic values are 95% confidence interval. *Observed
IC.sub.50 significantly lower (p < 0.05) than the expected IC50
and CI = 2.50 indicating synergy of the five-component
formulation.
Conclusion--With a CI=2.50, F2 unexpectedly inhibited MPO 2.5-times
more effectively than the sum of its components indicating synergy
of the combination. F1 did not inhibit MPO more or less effectively
than the sum of its components representative of an additive
response.
Example 3
Acute Nitric Oxide Production by a Copper Chlorophyllin/Spearmint
Oil Formulation
[0202] A liquid chlorophyllin/spearmint oil, shown below, was
tested to assess the acute, biosynthesis of the NO biomarker
NO.sub.2 in saliva in subjects with normal plasma concentrations of
ADMA.
TABLE-US-00005 TABLE 5 Ingredients in Test Formulation 3 Formula 3
Ingredient [mg] [Active Fraction] Propylparaben [NF] 189 --
Methylparaben 476 -- Spearmint Aerial Parts Oil (active) 414 0.226
Sodium Copper Chlorophyllin (active) 1,419 0.774 Ingredients listed
were dissolved in 473.1 mL of purified water.
Methods--The assay of nitrite in saliva and nitrate in Test
Formulation 3 (F3) was conducted as described in Example 1. Color
development of NO strips was quantitated as described in Example 1.
Means and 95% confidence intervals were computed as described in
Example 1. During the morning period, subjects (n=5), with normal
plasma concentrations of ADMA, were instructed to consume 5 mL of
the F3 with 500 mL of water subsequent to developing the first,
pre-dose NO strip. At post-dosing times 30, 60 and 90 minutes, NO
strips were again developed for all subjects. Results--As seen in
FIG. 10, F3 unexpectedly produced an average, acute increase of 42%
relative to zero time at 30 min (p<0.05). The acute increase in
NO biomarker with F3 continued with a 22% increase relative to zero
time at 60 min (p<0.05) and returned to approximately 15% below
pre-test concentrations at 90 min. The blend of sodium copper
chlorophyllin and spearmint oil (aerial parts) produced an acute
increase in the salivary NO biomarker NO.sub.2, indicative of a
rapid biosynthesis of NO.
Example 4
Synergistic Inhibition of Myeloperoxidase Activity by a
Ten-Component Phytocomplex
[0203] A 10-component phytocomplex was tested for synergy in in
vitro inhibition of MPO. All chemicals, methods, samples, assay,
and calculations were performed as described in Example 2.
[0204] The PC10 test material was formulated by combining apple
fruit extract, bergamot fruit extract, blueberry fruit concentrate,
capsicum fruit, grape seed extract, grape skin extract, green tea
leaf extract, mangosteen pericarp extract, olive leaf extract, and
turmeric root & rhizome extract in a number of the ratios
beginning with 1:1:1:1:1:1:1:1:1:1 and increasing or decreasing the
relative amount of a component originally based upon antioxidant
activity and cost of ingredient to arrive at the
6:50:1:1:1:1:3:1:1:6 ratio exhibiting the best overall synergy.
This formulation served as the basis for testing MPO inhibition and
is listed in Table 6.
TABLE-US-00006 TABLE 6 Ten-component Phytocomplex (PC10) Observed
IC.sub.50 Relative Amount Test Material [.mu.g/mL] [F] Apple
fruit.dagger. 3.86 (3.19-4.66) 0.085 Bergamot fruit.dagger. 42.0
(28.5-61.7) 0.704 Blueberry fruit* 140 (92.4-212) 0.014 Capsicum
fruit.dagger. 34.5 (31.5-42.3) 0.014 Grape seed.dagger. 2.40
(1.81-3.18) 0.014 Grape skin.dagger. 34.5 (28.9-41.1) 0.014 Green
tea leaf.dagger. 0.883 (0.557-1.40) 0.042 Mangosteen 3.35
(2.46-4.56) 0.014 pericarp.dagger. Olive leaf.dagger. 8.82
(6.27-12.4) 0.014 Turmeric root & 95.5 (75.0-122) 0.085
rhizome.dagger. Combination Index Expected [Expected IC.sub.50
IC.sub.50/ [.mu.g/mL] Observed IC.sub.50] Phytocomplex 2.95*
(2.17-4.03) 10.00 3.39 (PC10) .dagger.extract/ *concentrate/ **
Phytocomplex PC10 contains relative amounts
Results--As seen in Table 6 above, the observed IC50 of the
ten-component phytocomplex was 2.95 .mu.g/mL, while the calculated,
expected IC50 value was 10.0 .mu.g/mL resulting in a combination
index (CI) of 3.39. Conclusion--The ten-component phytocomplex
(PC10) in the ratios of about 6:50:1:1:1:1:3:1:1:6 exhibited a
dramatic, unexpected increase 3.4-times the MPO-inhibitory activity
of its individual components thus exhibiting synergy of the
formulation.
Example 5
Clinical Assessment of PC10 in Normal and Pre-Diabetic Subjects
[0205] The clinical safety and efficacy of the PC10 formula was
determined in an open-label, observational clinical trial. The
study population included males and females between the ages of 18
to 72 inclusive exhibiting the following lipid variables: serum
triglycerides.gtoreq.150 mg/dl and/or serum low density lipoprotein
cholesterol (calculated).gtoreq.150 mg/dl. During the 12-week
study, subjects were assigned to one of three groups to receive,
respectively, 500, 750, or 1000 mg of PC10 to be taken as 2, 3, or
4 capsules taken once daily with the evening meal.
[0206] Subjects were instructed to maintain their current
lifestyles including diet, exercise, and mind body spirit practices
without change during trial participation. Subjects were also
instructed not to make changes to their current prescription,
non-prescription medications, medical foods or nutritional
supplements while on the study.
[0207] At one, two and three months, blood was drawn for analysis
including complete blood count (CBC), complete metabolic panel
(CMP), fasting lipid panel including total cholesterol,
triglycerides, HDLc, LDLc, oxLDL, MPO, PAI-1 and HbA1c.
[0208] During this 12-week trial, there were no reported adverse
events related to the PC10 in the 500, 750, or 1000 mg/day groups
(N=35). Efficacy was assessed only for the potential commercial
formulation of 500 mg PC10 (n--11).
[0209] Table 7 summarizes the changes (Initial--3 Months) in median
lipid variables in subjects consuming 500 mg daily of the PC10
formulation over three months. Statistically significant changes
were noted in Total Cholesterol, Total Cholesterol/HDL ration,
LDL-c (calculated), Apo B and non-HDL for the group of eleven. The
change in Total Cholesterol of 7% and LDL-c (calculated) of 10% are
considered clinically meaningful.
[0210] Additionally, a subgroup analysis was conducted for subjects
with a HbA1C.gtoreq.5.5% (all of whom were insulin resistant with
HOMA scores greater than 2). Statistically significant changes were
noted in Total Cholesterol, Total Cholesterol/HDL ration, LDL-c
(calculated), Apo B, LDL-c (calculated)/HDL ratio, oxLDL, oxLDL/HDL
ratio, non-HDL, Triglycerides, TG/HDL ratio and plasminogen
activator inhibitor-1 (PAI-1) in this group of eight subjects. The
change in Total Cholesterol of 10%, LDL-c (calculated) of 10%,
oxLDL of 19%, TG of 27% and PAI-1 of 37% are clinically meaningful
and demonstrate the broad spectrum of action of the PC10
formulation.
TABLE-US-00007 TABLE 7 Median Changes in Lipid Biomarkers in all
Subjects and Subset of Subjects with Elevated HbA1c Consuming 500
mg Daily of The PC10 Formulation over Three Months Total (n = 11)
HbA1c >5.4 (n = 8).dagger. Median Change Median Change Variable
(% Change) P* (% Change) P* Weight 0.0 (0.0) NS 1.0 (0.0) NS Total
-20 (-7.0) 0.003 -23 (-10) 0.008 Cholesterol HDL -10 (-3.0) NS 4.0
(8.0) NS Cholesterol/HDL -2.0 (-26) 0.024 -2.5 (-45) 0.016 LDL -19
(-10) 0.012 -21 (-10) 0.031 oxLDL -6.0 (-10) NS -14 (-19) 0.047
APOB -4.0 (-3.0) 0.037 -7.0 (-4.0) 0.016 oxLDL/HDL -0.2 (-17) NS
-0.3 (-25) 0.039 Non-HDL -16 (-7.0) 0.007 -21 (-11) 0.008
Triglycerides -24 (-9.0) NS -35 (-27) 0.039 LDL/HDL -0.3 (-7.0) NS
-0.4 (-10) 0.031 PAI-1 -4.0 (-20) NS -7.0 (-37) 0.047 *P-values
were computed using the log-normal distribution of the ratio of
change from baseline to 12 weeks using the Wilcoxon Signed Rank
test of the median. The Null Hypothesis assumed a mean change from
baseline of zero. NS = nonsignificant (P > 0.05)
.dagger.Subgroup of subjects selected with HbA1c greater than 5.4;
bolding highlights subgroup differences.
[0211] In both groups, the effect of PC10 on oxLDL levels was
significant. For the group overall, there was a 10% reduction that
nearly placed subjects at completion in the low risk group. The
subgroup began the trial at moderate risk and had improved with a
fall to the low risk group by completion. This reduction in an
important risk factor for the development of coronary artery
disease offers an additional opportunity to promote healthy aging.
The antioxidant components of the formula function to assist in
lowering oxLDL levels and promote a healthy cholesterol metabolism
to offer organ system protection.
Example 6
Synergistic Inhibition of Myeloperoxidase Activity by a
Nine-Component Phytocomplex
[0212] A trial was conducted to assess the ability of a 9-component
phytocomplex (PC9p) to exhibit synergy in the in vitro inhibition
of MPO. All Chemicals, Methods and Calculations were performed as
described in Example 2. PC9p Test material--The PC9p test material
was formulated as a reduced version of PC10 lacking only the
bergamont orange fruit extract (Table 8).
TABLE-US-00008 TABLE 8 Nine-component Phytocomplex (PC9) Observed
IC50 Relative Amount Test Material [.mu.g/mL] [F] Apple
fruit.dagger. 3.86 (3.19-4.66) 0.286 Blueberry fruit* 140
(92.4-212) 0.048 Capsicum fruit.dagger. 34.5 (31.5-42.3) 0.048
Grape seed.dagger. 2.40 (1.81-3.18) 0.048 Grape skin.dagger. 34.5
(28.9-41.1) 0.048 Green tea leaf.dagger. 0.883 (0.557-1.40) 0.143
Mangosteen 3.35 (2.46-4.56) 0.048 pericarp.dagger. Olive
leaf.dagger. 8.82 (6.27-12.4) 0.048 Turmeric root & 95.5
(75.0-122) 0.286 rhizome.dagger. Combination Index Expected
[Expected IC.sub.50 IC.sub.50/ [.mu.g/mL] Observed IC.sub.50]
Phytocomplex 1.95* (1.19-4.03) 3.68 1.89 (PC9) .dagger.extract/
*concentrate/ ** Phytocomplex PC9 contains relative amounts
*Observed IC50 of the PC9p formulation is significantly (p <
0.05) less than the Expected IC50 with a CI = 1.89 indicating
synergy.
Conclusion--The nine-component phytocomplex (PC9p) exhibited an
unexpected increase 1.9-times the MPO-inhibitory activity of its
individual components thus exhibiting synergy of the formulation
(Table 8).
Example 7
Synergistic Inhibition of Myeloperoxidase Activity by
Four-Component Phytocomplexes
[0213] Two 4-component phytocomplexes, differing only in the ratios
of their components were tested, to exhibit synergy in the in vitro
inhibition of MPO. All Chemicals, Methods and Calculations were
performed as described in Example 2.
PC9p Test material--The PC4x test combinations were formulated as a
further reduced version of PC10 as described in Tables 9 and
10.
TABLE-US-00009 TABLE 9 CI of Four-Component Phytocomplex (PC4.1)
for Myeloperoxidase Inhibition Observed IC.sub.50.dagger. Fraction
of Test Material [.mu.g/mL] PC4.1 Formulation Apple Fruit Extract
3.86 (3.19-4.66) 0.25 Grape Seed Extract 2.40 (1.81-3.18) 0.25
Green Tea Leaf 0.883 (0.557-1.40) 0.25 Extract Olive Leaf Extract
8.82 (6.27-12.4) 0.25 Combination Index Expected [Expected
IC.sub.50 IC.sub.50/ [.mu.g/mL] Observed IC.sub.50] PC4.2 1.08*
(0.778-1.49) 2.08 1.93 .dagger.Parenthetic values are 95%
confidence interval. *Observed IC50 significantly lower (p <
0.05) than the expected IC50 and CI = 1.93 indicating synergy of
the four-component formulation.
[0214] The four-component phytocomplex (PC4.1) in the ratios of
about 1:1:1:1 exhibited a dramatic, unexpected increase nearly
2-times the MPO-inhibitory activity of its individual components
thus exhibiting synergy of the formulation (Table 9).
TABLE-US-00010 TABLE 10 CI of a Four-Component Phytocomplex (PC4.2)
for Myeloperoxidase Inhibition Observed IC.sub.50.dagger. Fraction
of Test Material [.mu.g/mL] PC4.2 Formulation Apple Fruit Extract
3.86 (3.19-4.66) 0.55 Grape Seed Extract 2.40 (1.81-3.18) 0.09
Green Tea Leaf 0.883 (0.557-1.40) 0.27 Extract Olive Leaf Extract
8.82 (6.27-12.4) 0.09 Combination Index Expected [Expected
IC.sub.50 IC.sub.50/ [.mu.g/mL] Observed IC.sub.50] PC4.2 0.42*
(0.29-0.62) 2.05 4.83 .dagger.Parenthetic values are 95% confidence
interval. *Observed IC50 significantly lower (p < 0.05) than the
expected IC50 and CI = 4.83 indicating synergy of the
four-component formulation.
[0215] The four-component phytocomplex (PC4.2) in the ratios of
about 6:1:3:1 exhibited a dramatic, unexpected increase nearly
5-times the MPO-inhibitory activity of its individual components
thus exhibiting synergy of the formulation (Table 10).
Example 8
Salivary Nitrite Levels after Oral Administration of Prototype and
Commercial Nitric Oxide Generating Products in Healthy
Volunteers
[0216] Saliva nitrite production levels in healthy subjects were
evaluated after taking commercial products containing putative NO
generating actives. In the study, eight subjects between the ages
of 18 to 72 performed a Berkeley Test.RTM. nitric oxide biomarker
strip (NOBS) test early in the morning to determine their baseline
level of salivary NO.sub.2. If the baseline score was below 5 on
the Visual scale, the subjects then proceeded to take one of the
test articles listed in Table 11. After consumption of the test
article, NOBS were developed at 30, 60, 90, 120 and 240
minutes.
TABLE-US-00011 TABLE 11 Description of the 15 Test Articles Used
Lot Serving Test Article Vendor Product Code Number (g) ProArgi9
Synergy 72840 10.5 Purple Wave ProArgi9 P1.sup.a Synergy 11.1
Niteworks Herbalife 3150US 475562B19 10.0 BeetElite Neogenis NA
17115 10.0 NO3 Nutrabolt 101742 0502C5 Chrome.sup.b L-Arginine
Elements of LPLUS-01 151001 12.7 Plus Health Care LLC L-Arginine
Nuvo Soma NA U1510826 10.6 Cardio Labs Power Perfusia-SR Thome
LSA52508 316622 Research L-Arginine Fenix 9015400288 47236300 10.0
Complete Nutrition Arginine Sante Global NA 15371 10.3 Infusion
Arginext.sup.c High Desert NA NA 13.5 Heart Inst. Nitroxyl
Nitrosolution XOOOMTPKU 1502077 LRG9 NA LRG912-BX30- 1420302 12.0
V01 Neo40 Neogenis NA 142591S 1.4 Arginine Dr. Boger 8104519792
15513 8 Cardio.sup.d .sup.aProArgi9 P1 contains FCC beet root
powder (lot 45). .sup.b1250 mg of arginine nitrate per serving
(approximately 919 mg of arginine and 331 mg of nitrate based on
molecular weight). .sup.cDr. Siva Arunasalam test article.
.sup.dDr. Rainer Boger test article.
Berkeley Nitric Oxide Biomarker Strips (NOBS)--NOBS were developed
and immediately scored with Visual scoring card. Subjects with
access to Argus app scored NOBS immediately. Subject (s) without
access to Argus app submitted NOBS to the study monitor for
scoring. Argus readings [units] were converted to .mu.M Nitrite
with an equation (y=0.0027x2+1.2755x+37.805; R2=0.9994) that
described a 2nd degree polynomial relationship between Argus units
and NOBS color development.
[0217] 10 of the 15 test materials contained arginine or citrulline
as a source of salivary NO.sub.2, 4 contained only NO.sub.3 as the
source of salivary NO.sub.2, and ProArgi9 P1 contained both the
amino acids and red beet root as potential sources of salivary
NO.sub.2 (Table 12). The complete compositions of ProArgi9 Purple
Wave and ProArgi9 P1 are presented in Tables 13 and 14,
respectively.
Arginine/Citrulline Content of Test Materials--Arginine or
citrulline content of the test materials was obtained from labels
on the commercial products. All commercial products were
administered per serving instructions on the container.
Determination of Nitrate and Nitrite in Test
Articles--Nitrate/Nitrite (NOx) fluorometric assays were performed
according to the manufacturer's instructions (Cayman Chemicals,
Item No. 780051, Ann Arbor, Mich.). Results were tabulated as mg
NOx/serving. Pharmacokinetic Parameters and Calculations--The
pharmacokinetic parameter of area-under-the-curve (AUC) was
computed using the trapezoidal method from 0 to 240 minutes. The
extent of NO.sub.2 increase relating to product administration was
computed using the ratio of AUC.sub.(0-240)/eAUC, where eAUC
represented the endogenous salivaryNO2 and was computed using
C.sub.0.times.240 min. Thus, AUC.sub.(0-240)/eAUC represented the
systemic NO increase provided by the product. If the 95% Confidence
Interval (CI) of the AUC.sub.(0-240)/eAUC included a value<1.0,
it was assumed that the test material had no systemic impact on NO
during the 240 minutes of the observation period.
TABLE-US-00012 TABLE 12 Arginine, Citrulline, Nitrate and Nitrite
Content of the 15 Test Articles Nitrate Nitrite Serving Arginine
Citrulline (mg/ (mg/ Test Article (g) (g/serving) (g/serving)
serving) serving) ProArgi9 Purple 10.5 5.1 0.044 <0.02 <0.02
Wave ProArgi9 P1 11.1 5.1 0.005 76.7 <0.02 Niteworks 10.0
5.2.sup.a <0.02 <0.02 BeetElite 10.0 200 6.67 NO3 Chrome 2.10
307 <0.03 L-Arginine Plus 12.7 5.1 1.01 <0.03 <0.03
L-Arginine 11.6 5.0 0.2 <0.03 <0.03 Cardio Power Perfusia-SR
1.39 1.0 <0.03 <0.03 L-Arginine 10.0 5.0 1.0 <0.03
<0.03 Complete Arginine Infusion 10.3 5.0 1.0 <0.03 <0.03
Arginext.sup.d 13.5 10.0.sup.a <0.02 <0.02 Nitroxyl 0.657
0.25 1.21 <0.03 LRG9 12.0 5.0 1.0 <0.03 <0.03 Neo40 1.4
1.49 9.10 Arginine Cardio 8.0 7.58.sup.a <0.02 <0.02
.sup.aProprietary blend of L-Arginine/L-Citrulline
TABLE-US-00013 TABLE 13 Composition of ProArg9 Purple Wave Amount
Ingredient (mg) ProArgi-9 Plus Vitamin Base 614 Red Grape
Polyphenol Extract [ExGrape(TM) red wine extract] 50 Grape Skin
Extract/Vitis vinifera 75 Watermelon Whole Fruit Extract [20%
Citrulline]/Citrullus 220.1 lanatus, Watermelon Powdered Extract,
20% Vitamin C (ascorbic acid) [100%, fine powder] 75 Vitamin B12
(cyanocobalamin) [1%, trituration] 0.9 Folic Acid [10%,
trituration] 3 Vitamin B6 (pyridoxine hydrochloride) [82% B6] 3
Silicon Dioxide [Syloid .RTM. 244] 50 D-Ribose 100 Vitamin D3
(cholecalciferol) [100,000 IU/g, 100 SD/S] 37 L-Arginine [granular]
5100 Pomegranate Fruit Juice Concentrate/Punica granatum 500 Stevia
Leaf Extract/Stevia rebaudiana 66 Natural Citrus Sweetener
[CitriSweet(TM)] 430 Xylitol [bulk] 500 Citric Acid 2500 Citrus
Blend Natural Flavor [WONF] 85 Malic Acid 500 Huckleberry Natural
Flavor 85 Silicon Dioxide [Syloid .RTM. 244] 120
TABLE-US-00014 TABLE 14 Composition of ProArgi9P1 Ingredient
ProArgi9P1 (mg) L-Arginine 5100 Citric Acid 2000 Red Beet Root 2000
Natural Citrus Sweetner 430 Malic Acid 400 Pomogrenate Fruit Juice
Concentrate 375 Silicon Dioxide 170 Thiamin (B1) (Thiamine
mononitrate) 110 Calcium Ascorbate 95 Citrus Blend Natural Flavor
85 Huckleberry Natural Flavor 85 Magnesium Oxide 84 Stevia Leaf
Extract* 66 Apple Fruit Extract 30 Watermelon Whole Fruit Extract
23 Vitamin D3 (Cholecalciferol) 2.4 Green Tea Leaf Extract 15
D-Ribose 10 Grape Skin Extract 5.0 Red Grape Polyphenol Extract 5.0
Grape Seed Extract 5.0 Olive Leaf Extract 5.0 Folic Acid 3.0
Vitamin B6 2.9 Vitamin B12 0.345
TABLE-US-00015 TABLE 15 AUC and Relative Increases in AUC for the
11 Test Articles Containing Arginine or Citrulline
AUC.sub.(0-24).dagger. AUC.sub.(0-24)/ [mmol-
AUC.sub.(0-24).dagger. AUC/ eAUC Test Article min/L] (95% CI)
eAUC.dagger..dagger. (95% CI) ProArgi9 Purple 25.9 20.3-33.2
1.23.sup.a 1.03-1.47 Wave ProArgi9 P1 65.7 41.4-105 3.19.sup.a
1.99-5.11 Niteworks 22.4 20.1-24.8 1.11 0.98-1.24 L-Arginine Plus
22.3 19.2-25.9 1.06 0.86-1.30 L-Arginine Cardio 21.1 19.6-22.7 1.00
0.92-1.10 Power Perfusia 22.9 20.9-25.2 0.96 0.77-1.19 L-Arginine
21.1 19.7-22.7 1.07 0.98-1.17 Complete Arginine Infusion 21.2
19.9-22.7 1.02 0.92-1.13 Arginext 22.0 20.0-24.2 1.03 0.99-1.07
LRG9 24.4 18.8-31.8 1.14 0.92-1.42 Arginine Cardio 20.9 19.7-22.2
1.02 0.97-1.07 .dagger.Values are means of 8 independent
observations; parenthetic values are 95% confidence limits
.dagger..dagger.The extent of NO2 increase relating to product
administration was computed using the ratio of
AUC.sub.(0-240)/eAUC, where eAUC represented the endogenous
salivaryNO2 and was computed using C.sub.0 .times. 240 min. Thus,
AUC.sub.(0-240)/eAUC represented the systemic NO increase provided
by the product. .sup.aSignificant increase over endogenous NO2.
[0218] ProArgi9 P1 produced the greatest relative increase in
salivary NO.sub.2 with an AUC.sub.(0-240)/eAUC of 3.19 or a 3-fold
increase in the endogenous level of NO.sub.2. This effect was
likely enhanced by the NO.sub.3 content of ProArgi9P1 combined with
MPO-inhibiting polyphenols. The only test material without NO.sub.3
that produced a significant (P<0.05) increase over endogenous
NO.sub.2 was ProArgi9 Purple Wave, which demonstrated an enhanced
systemic production of NO relative to formulations that do not
contain MPO-inhibiting polyphenols.
TABLE-US-00016 TABLE 16 AUC and Relative Increases in AUC for the 5
Test Articles Containing NO.sub.3 AUC.sub.(0-24) .dagger. AUC/
[mmol- AUC.sub.(0-24) .dagger. AUC/ eAUC Test Article min/L] (95%
CI) eAUC.dagger..dagger. (95% CI) ProArgi9 P1 65.7 41.4-105
3.19.sup.a 1.99-5.11 BeetElite 102 61.3-169 4.40.sup.a 2.51-7.71
NO3 Chrome 107 60.5-189 5.00.sup.a 2.84-8..80 Nitroxyl 20.2
19.4-21.1 1.00 0.94-1.06 Neo40 24.3 20.6-28.6 1.14 0.96-1.35
.dagger. Values are means of 8 independent observations;
parenthetic values are 95% confidence limits .dagger..dagger.The
extent of NO2 increase relating to product administration was
computed using the ratio of AUC.sub.(0-240)/eAUC, where eAUC
represented the endogenous salivaryNO2 and was computed using
C.sub.0 .times. 240 min. Thus, AUC.sub.(0-240)/eAUC represented the
systemic NO increase provided by the product. .sup.aSignificant
increase over endogenous NO2.
Of the five test materials, only ProArgi9 P1, BeetElite and
NO.sub.3 Chrome produced significant (P<0.05) increases above
endogenous NO.sub.2 as represented by the lower limit of the 95% CI
of the AUC.sub.(0-240)/eAUC>1.0. The NO.sub.3 content of these
three products, however, varied greatly, 76.7, 200, and 307,
respectively, for ProArgi9 P1, BeetElite, and NO.sub.3 Chrome. To
determine the effect of formulation on the increase in salivary
NO.sub.2, the AUC/eAUC ratio was adjusted for NO.sub.3 dose (Table
17).
TABLE-US-00017 TABLE 17 NO3 Dose-adjusted Increases in AUC/eAUC for
Test Materials Producing Salivary NO.sub.2 Above Endogenous Levels
Test Article AUC/eAUC.dagger.# (AUC/eAUC)/g NO3.dagger..dagger.
ProArgi9 P1 3.19 42 (26-68).sup.a BeetElite 4.40 22 (13-39).sup.b
NO3 Chrome 5.00 16 (9.3-29).sup.b .dagger.Values are means of 8
independent observations. #The extent of NO2 increase relating to
product administration was computed using the ratio of
AUC.sub.(0-240)/eAUC, where eAUC represented the endogenous
salivaryNO2 and was computed using C.sub.0 .times. 240 min. Thus,
AUC.sub.(0-240)/eAUC represented the systemic NO increase provided
by the product. .dagger..dagger.Ratio adjusted for 76.7, 200 and
307 mg dosing respectively for ProArgi9 P1, BeetElite and NO3
Chrome; parenthetic value is 95% confidence interval.
.sup.a,bUncommon superscripts indicate significant differences (P
< 0.05).
When adjusted for NO.sub.3 dose, the ProArgi9 P1 formulation was
1.9- and 2.6-fold more active than BeetElite and NO.sub.3 Chrome,
respectively. It can be inferred from this example that a
formulation containing MPO-inhibiting polyphenols can enhance
systemic production of NO from similar doses of NO.sub.3 relative
to formulations that do not contain MPO-inhibiting polyphenols.
Example 9
Formulations for Enhancing Nitric Oxide Production from Dietary
NO.sub.3/NO.sub.2
[0219] Based upon results presented, the following formulations
would be expected to perform as effectively as PA9-PW or
ProArgi9P1.
TABLE-US-00018 TABLE 18 Composition of PA (NO+ (MY)) Ingredient
(mg) L-Arginine 5100 Citric Acid 2200 Red Beet Root (Nitrate 2%)
2000 Malic Acid 450 Natural Citrus Sweetener (CitriSweet .TM.) 430
Pomegranate Fruit Juice Concentrate 375 Silicon Dioxide (Syloid
.RTM. 244) 170 Thiamin (B1) (thiamie mononitrate) 110 Calcium
Ascorbate (83% vit C, 9% Ca) 95 Citrus Blend Natural Flavor (WONF)
90 Natural Fresh Fruit Cherry 90 Magnesium Oxide (60% Mg, powder)
84 Stevia Leaf Extract 66 Apple Fruit Extract (75% polyphenols) 30
Watermelon Whole Fruit Extract (20% Citru 23 Green Tea Leaf Extract
(80% decaffeinate) 15 D-Ribose 10 Grape Skin Extract 5.0 Red Grape
Polyphenol Extract (ExGrape .TM.) 5.0 Grape Seed Extract
(MegaNatural .RTM.) 5.0 Olive Leaf Extract (12%, 7:1) 5.0 Folic
Acid (10% trituration) 3.0 Vitamin B6 (pyridoxine hydrochloride)
82% 2.9 Vitamin D3 (cholecalciferol) (100,000 IU/g, 2.4 Vitamin B12
(cyanocobalamin) 0.345
TABLE-US-00019 TABLE 19 Composition of ProArg9P2 Ingredient (mg)
L-Arginine 5100 Citric Acid 2000 Red Beet Root (about 2% NO3) 2000
Natural Citrus Sweetner 430 Malic Acid 400 Pomogrenate Fruit Juice
Concentrate 375 Silicon Dioxide 170 Thiamin (B1) (Thiamine
mononitrate) 110 Calcium Ascorbate 95 Citrus Blend Natural Flavor
85 Huckleberry Natural Flavor 85 Magnesium Oxide 84 Stevia Leaf
Extract* 66 Apple Fruit Extract 30 Watermelon Whole Fruit Extract
23 Vitamin D3 (Cholecalciferol) 2.4 Green Tea Leaf Extract 15
D-Ribose 10 Grape Skin Extract 5.0 Red Grape Polyphenol Extract 5.0
Grape Seed Extract 5.0 Olive Leaf Extract 5.0 Folic Acid 3.0
Vitamin B6 2.9 Vitamin B12 0.345
TABLE-US-00020 TABLE 20 Composition of Biome NO+ Ingredient (mg)
Red Grape Polyphenol Extract 5.0 Apple Fruit Extract [75%
polyphenols]/Malus pumila 30 Grape Seed Extract [MegaNatural
.RTM.]/Vitis vinifera 5.0 Grape Skin Extract/Vitis vinifera 40
Green Tea Leaf Extract [80%, decaffeinated]/Camellia sinensis 15
L-Arginine [granular] 1000 Olive Leaf Extract [12%, 7:1]/Olea
europaea 5.0 Pomegranate Fruit Juice Concentrate/Punica granatum
450 Red Beet Root (Nitrate 2%) (KR)/Beta vulgaris 3000 Stevia Leaf
Extract/Stevia rebaudiana 70 Natural Citrus Sweetener
[CitriSweet(TM)] 450 Watermelon Whole Fruit Extract [20%
Citrulline]/Citrullus 23 lanatus Watermelon Powdered Extract, 20%
Xylitol [bulk] 500 Vitamin C (ascorbic acid) [100%, fine powder] 75
Citric Acid 1250 Citrus Blend Natural Flavor [WONF] 100 Vitamin B12
(cyanocobalamin) [1%, trituration] 0.9 Folic Acid [10%,
trituration] 3.0 L-Glutamine 1000 Inulin (chicory root extract) [HD
food grade] 2021 Magnesium Oxide [60% Mg, powder] 84 Malic Acid 275
Natural Fresh Fruit Cherry 89 Vitamin B6 (pyridoxine hydrochloride)
[82% B6] 3.0 Silicon Dioxide [Syloid .RTM. 244] 150 Thiamin (B1)
(thiamine mononitrate) [91% B1] 110 D-Ribose 30 Vitamin D3
(cholecalciferol) [100,000 IU/g, 100 SD/S] 16
[0220] Methods for the production of these formulations and uses
have been described. It will be readily apparent to those skilled
in the art, however various changes and modifications of an obvious
nature may be made without departing from the spirit of the
invention, and all such changes and modifications are considered to
fall within the scope of the invention as defined by the appended
claims. Such changes and modifications would include, but not be
limited to, the incipient ingredients added to affect the capsule,
tablet, powder, lotion, food, powder, or bar manufacturing process
as well as vitamins, flavorings, and carriers. Other such changes
or modifications would include the use of herbs or other botanical
products containing the combinations of the preferred embodiments
disclosed above. Many additional modifications and variations of
the embodiments described herein may be made without departing from
the scope, as is apparent to those skilled in the art. The specific
embodiments described herein are offered by way of example
only.
* * * * *