U.S. patent application number 14/255431 was filed with the patent office on 2014-08-14 for treating inflammation with a combination of an ellagitannin and hydrogen peroxide.
This patent application is currently assigned to LiveLeaf, Inc.. The applicant listed for this patent is LiveLeaf, Inc.. Invention is credited to ALEXANDER L. HUANG, GIN WU.
Application Number | 20140227367 14/255431 |
Document ID | / |
Family ID | 47362044 |
Filed Date | 2014-08-14 |
United States Patent
Application |
20140227367 |
Kind Code |
A1 |
HUANG; ALEXANDER L. ; et
al. |
August 14, 2014 |
TREATING INFLAMMATION WITH A COMBINATION OF AN ELLAGITANNIN AND
HYDROGEN PEROXIDE
Abstract
The teachings provided herein generally relate to site-activated
binding systems that selectively increase the bioactivity of
phenolic compounds at target sites. More particularly, the systems
taught here include a phenolic compound bound to a reactive oxygen
species, wherein the phenolic compound and the reactive oxygen
species react at a target area in the presence of an oxidoreductase
enzyme.
Inventors: |
HUANG; ALEXANDER L.; (Menlo
Park, CA) ; WU; GIN; (San Rafael, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
LiveLeaf, Inc. |
San Carlos |
CA |
US |
|
|
Assignee: |
LiveLeaf, Inc.
San Carlos
CA
|
Family ID: |
47362044 |
Appl. No.: |
14/255431 |
Filed: |
April 17, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14203517 |
Mar 10, 2014 |
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14255431 |
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13135124 |
Jun 24, 2011 |
8722040 |
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14203517 |
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Current U.S.
Class: |
424/616 |
Current CPC
Class: |
A61P 17/00 20180101;
A61P 1/00 20180101; Y02A 50/30 20180101; A61P 1/12 20180101; A61P
39/02 20180101; A61K 36/82 20130101; A61P 1/04 20180101; A61P 1/10
20180101; A61P 1/14 20180101; A61K 31/365 20130101; A61K 36/185
20130101; A61K 36/22 20130101; A61K 31/192 20130101; A61P 37/08
20180101; A61K 31/366 20130101; A61P 1/08 20180101; A61K 31/05
20130101; A61K 31/35 20130101; A61P 43/00 20180101; A61K 33/40
20130101; A61P 3/02 20180101; A61P 29/00 20180101; A61P 31/04
20180101; A61K 31/353 20130101; A61K 31/7032 20130101 |
Class at
Publication: |
424/616 |
International
Class: |
A61K 33/40 20060101
A61K033/40; A61K 31/192 20060101 A61K031/192; A61K 36/82 20060101
A61K036/82; A61K 31/353 20060101 A61K031/353; A61K 31/366 20060101
A61K031/366; A61K 36/185 20060101 A61K036/185 |
Claims
1. A method of treating an inflammation in a subject, comprising:
administering an effective amount of a binding system to a subject
having an inflammation, the binding system comprising: an
ellagitannin; and, hydrogen peroxide; wherein, the hydrogen
peroxide is combined with the ellagitannin at a weight ratio that
ranges from about 1:1000 to about 10:1 of the an extract to the
hydrogen peroxide; and, the administering inhibits, or ameliorates
the symptoms of, the inflammation of the subject.
2. The method of claim 1, wherein the administering of the binding
system includes obtaining the binding system as a pharmaceutical
formulation comprising the binding system of claim 1 and a
pharmaceutically acceptable excipient.
3. The method of claim 1, wherein the ellagitannin is a
punicalagin.
4. The method of claim 1, wherein the binding system further
comprises a catechin.
5. The method of claim 1, wherein the binding system further
comprises gallic acid, epigallic acid, or a combination
thereof.
6. The method of claim 1, wherein the ellagitannin is an extract of
pomegranate.
7. The method of claim 4, wherein the catechin is an extract of
green tea.
8. The method of claim 1, wherein the weight ratio of the
ellagitannin to the hydrogen peroxide ranges from about 1:1 to
about 1:50.
9. The method of claim 1, wherein the administering further
comprises obtaining a kit comprising the binding system, wherein
the kit comprises a dry form of the binding system, as well as
instructions for diluting the binding system for administration
with suggested dilution factors for various target sites.
10. A method of inhibiting a gastrointestinal-mediated autoimmune
response in a subject, comprising: administering an effective
amount of a binding system to a subject having a
gastrointestinal-mediated autoimmune activity, the binding system
comprising: an ellagitannin; and, hydrogen peroxide; wherein, the
hydrogen peroxide is combined with the ellagitannin at a weight
ratio that ranges from about 1:1000 to about 10:1 of the an extract
to the hydrogen peroxide; and, the administering inhibits, or
ameliorates the symptoms of, the gastrointestinal-mediated
autoimmune activity in the subject.
11. The method of claim 10, wherein the administering of the
binding system includes obtaining the binding system as a
pharmaceutical formulation comprising the binding system of claim 1
and a pharmaceutically acceptable excipient.
12. The method of claim 10, wherein the ellagitannin is a
punicalagin.
13. The method of claim 10, wherein the binding system further
comprises a catechin.
14. The method of claim 10, wherein the binding system further
comprises gallic acid, epigallic acid, or a combination
thereof.
15. The method of claim 10, wherein the ellagitannin is an extract
of pomegranate.
16. The method of claim 13, wherein the catechin is an extract of
green tea.
17. The method of claim 10, wherein the weight ratio of the
ellagitannin to the hydrogen peroxide ranges from about 1:1 to
about 1:50.
18. The method of claim 10, wherein the administering further
comprises obtaining a kit comprising the binding system, wherein
the kit comprises a dry form of the binding system, as well as
instructions for diluting the binding system for administration
with suggested dilution factors for various target sites.
19. A composition, comprising: an ellagitannin; hydrogen peroxide;
and, a pharmaceutically acceptable carrier; wherein, the hydrogen
peroxide is combined with the ellagitannin at a weight ratio that
ranges from about 1:1000 to about 10:1 of the ellagitannin to the
hydrogen peroxide.
20. The composition of claim 19, wherein the ellagitannin is a
punicalagin.
21. The composition of claim 19, wherein the composition further
comprises a catechin.
22. The composition of claim 19, wherein the composition further
comprises gallic acid, epigallic acid, or a combination
thereof.
23. The composition of claim 19, wherein the ellagitannin is an
extract of pomegranate.
24. A formulation comprising the composition of claim 19 and a
pharmaceutically acceptable excipient.
25. The formulation of claim 24, wherein the weight ratio of the
ellagitannin to the hydrogen peroxide ranges from about 1:1 to
about 1:50.
26. A kit for generating the composition of claim 19, wherein the
kit comprises a dry form of the punicalagin and a dry form of a
hydrogen peroxide generating material, as well as instructions for
mixing the components for administration and suggested dilution
factors for various target sites.
27. A formulation, comprising: a punicalagin; hydrogen peroxide;
and, a pharmaceutically acceptable excipient; wherein, the hydrogen
peroxide is combined with the punicalagin at a weight ratio that
ranges from about 1:1000 to about 10:1 of the an extract to the
hydrogen peroxide.
28. The formulation of claim 27, wherein the formulation further
comprises a catechin.
29. The formulation of claim 27, wherein the punicalagin is an
extract of pomegranate.
30. A kit for generating the formulation of claim 27, wherein the
kit comprises a dry form of the punicalagin, a dry form of a
hydrogen peroxide generating material, and the excipient, as well
as instructions for mixing the components for administration and
suggested dilution factors for various target sites.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. application Ser.
No. 14/203,517, filed Mar. 10, 2014, which is a continuation of
U.S. application Ser. No. 13/135,124, filed Jun. 24, 2011, each of
which is hereby incorporated by reference in its entirety.
BACKGROUND
[0002] 1. Field of the Invention
[0003] The teachings provided herein relate to site-activated
binding systems that selectively increase the bioactivity of
phenolic compounds at target sites.
[0004] 2. Description of Related Art
[0005] Some phenolic compounds, such as the polyphenols, are
considered beneficial for use as antioxidants in animals, such as
humans, due to their ability to scavenge unwanted reactive oxygen
species in vivo. Such reactive oxygen species can include, for
example, singlet oxygen, peroxynitrite, and hydrogen peroxide. This
ability to scavenge these reactive oxygen species can affect
cell-to-cell signaling, receptor sensitivity, inflammatory enzyme
activity and even gene regulation. An antioxidant molecule can, for
example, inhibit the oxidation of molecules and are characterized
as having a multiplicity of polar moieties that form bonds with
oxidizers such as hydrogen peroxide.
[0006] Nutritionists have long-recognized the unique health
benefits of "live" uncooked fruits and vegetables in the diet. The
main source of polyphenols for humans is currently dietary, since
they are found in a wide array of phytochemical-bearing foods. For
example, honey; most legumes; fruits such as apples, blackberries,
blueberries, cantaloupe, cherries, cranberries, grapes, pears,
plums, raspberries, and strawberries; and vegetables such as
broccoli, cabbage, celery, onion and parsley are rich in
polyphenols. red wine, chocolate, green tea, olive oil, argan oil,
bee pollen and many grains are sources of these compounds. It is
well known that many plant polyphenols ingested or otherwise
introduced to animal physiology vary greatly in bioavailability and
potency. Moreover, many examples of traditional medicines using
living or freshly harvested plant materials have only short lived
potency. In addition, all current extraction methods including
solvent, reflux heating, sonication, maceration and microwave
techniques disrupt intracellular structures, triggering mixing of
oxidoreductase enzymes with polyphenols. The polyphenols typically
oxidize in the process and have a tendency to autopolymerize or
complex indiscriminately with other extracted compounds, destroying
significant bioactive potential in a short period of time. Another
problem is that many medicinally useful polyphenol compounds also
have poor bioavailability. Oxidized polyphenols typically have
increased astringent binding activity but also have the tendency to
complex indiscriminately with body tissues, body fluids, or foods
in the digestive tract. In addition, another problem is that
bioactivation of the phenolic compounds requires reactive oxygen
species and, in some embodiments, the target site is an anaerobic
physiologic environment, and the phenolic compound has difficulty
activating.
[0007] As a result of at least the above, studies have failed to
demonstrate definitive health benefits from dietary supplementation
with antioxidants, such as polyphenols. Others have even shown
negative effects, including toxic effects from an excessive
ingestion of an antioxidant in an attempt to achieve the desired
effects. And, most studies, at best, have shown a low
bioavailability and rapid excretion of orally ingested antioxidant
polyphenol supplements from in vivo systems. As such, the art has
still not found an effective way to utilize the health improving
potential of these natural phenolic compounds.
[0008] One of skill would appreciate having a broad spectrum system
to bind compromised tissues, irritants and pathogens that includes
these seemingly desirable phenolic compounds, particularly a system
that (i) is stable, or at least substantially stable, for storage
or administration; (ii) selectively bioactivates the binding system
at a target site without significant indiscriminate complexing in
undesirable locations; (iii) functions as an astringent, an
antitoxin, an antimicrobial, an anti-inflammatory, an
anti-infectant, and the like, reacting with pathogens, their
virulence factors, pro-inflammatory compounds and damaged host
tissues; and, (iv) functions surprisingly well in small amounts on
dermal, mucosal, or in the GI tract tissue of an animal subject,
whether human or non-human, aeorobic or anaerobic environments, to
target and bind or exclude unwanted materials to treat health
conditions, maintain health, and supplement the health and
nutrition of the subject.
SUMMARY
[0009] The teachings provided herein generally relate to
site-activated binding systems that selectively increase the
bioactivity of phenolic compounds at a target site.
[0010] The teachings include a binding system that selectively
increases the bioactivity of phenolic compounds at a target site.
In some embodiments, the system can include a phenolic compound
component and a reactive oxygen species component. The phenolic
compound component can comprise a tannin having a molecular weight
ranging from about 500 Daltons to about 4000 Daltons; and, the
reactive oxygen species component can comprise hydrogen peroxide.
In some embodiments, the hydrogen peroxide can be releasably bound
to the tannin at a tannin:peroxide weight ratio (a molar weight
ratio) that ranges from about 1:1000 to about 10:1. In some
embodiments, the weight ratio of the tannin:peroxide ranges from
about 1:1 to about 1:50. And, in some embodiments, the binding
system is bioactivated at a target site having an oxidoreductase
enzyme that is expressed in response to a tissue damage. In these
embodiments, the phenolic compound component can bind to the target
site selectively. Moreover, in some embodiments, the binding system
contains no, or substantially no, unbound hydrogen peroxide prior
to the bioactivating at the target site. The teachings also include
a pharmaceutical formulation comprising the binding systems taught
herein and a pharmaceutically acceptable excipient.
[0011] In some embodiments, the binding molecule comprises a
hydrolysable tannin. In some embodiments, the binding molecule
comprises a condensed tannin. And, in some embodiments, the binding
molecule comprises a combination of a hydrolysable tannin and a
condensed tannin.
[0012] In some embodiments, the phenolic compound component
comprises a flavanol. In some embodiments, the phenolic compound
component comprises a catechin. And, in some embodiments, the
phenolic compound component comprises gallic acid, epigallic acid,
or a combination thereof.
[0013] The target site can be a damaged tissue of a subject. As
such, the teachings include a method of treating a damaged dermal,
mucosal, or gastrointestinal tissue. In some embodiments, the
method includes administering an effective amount of a binding
system taught herein to the damaged tissue of the subject. In some
embodiments, the binding system functions as an antitoxin when
bioactivated at the target site of the damaged tissue and assists
in the healing of the damaged tissue by inactivating toxic
compounds at the target site.
[0014] The teachings are also directed to a method of treating a
damaged dermal, mucosal, or gastrointestinal tissue. In some
embodiments, the method can comprise administering an effective
amount of a binding system taught herein to the damaged tissue of
the subject. The binding system can function as an antimicrobial
when bioactivated at the target site of the damaged tissue and
assist in the healing of the damaged tissue by inactivating
compounds that promote infection at the target site.
[0015] The teachings are also directed to a method of treating a
gastrointestinal condition. In some embodiments, the method can
comprise administering an effective amount of a binding system
taught herein to the gastrointestinal tract of the subject. The
binding system can function as an astringent, an anti-toxin, an
anti-inflammatory, or an antimicrobial, for example, when
bioactivated at the target site of the damaged tissue and assists
in the healing of the damaged tissue by inactivating compounds that
promote the condition at the target site.
[0016] The teachings are also directed to a method of treating
acute diarrhea in a subject. In some embodiments, the methods
comprise orally administering an effective amount of a binding
system taught herein to the subject. The binding system can
prevent, inhibit, or ameliorate a symptom of acute diarrhea in the
subject when compared to a second subject in a control group in
which the binding system was not administered. In some embodiments,
the symptom is selected from the group consisting of a stool score,
heartburn, indigestion, urgency of defecation, nausea, vomiting,
stomach pain, and bloating.
[0017] The teachings are also directed to a method of promoting
weight gain in a subject. In some embodiments, the method comprises
orally administering an effective amount of a binding system taught
herein to the subject as a supplement to the diet of the subject.
The binding systems can increase the feed conversion ratio of the
subject when compared to a second subject in a control group in
which the binding system was not administered.
[0018] The teachings are also directed to a method of treating
irritable bowel syndrome in a subject. In some embodiments, the
method comprises orally administering an effective amount of a
binding system taught herein to the subject. The binding system can
prevent, inhibit, or ameliorate the symptoms of irritable bowel
syndrome in the subject when compared to a second subject in a
control group in which the binding system was not administered. In
some embodiments, the symptom is selected from the group consisting
of a stool score, heartburn, indigestion, urgency of defecation,
nausea, vomiting, stomach pain, and bloating.
[0019] The teachings are also directed to a method of treating an
inflammatory bowel disease in a subject. In some embodiments, the
method comprises orally administering an effective amount of a
binding system taught herein to the subject. The binding system can
prevent, inhibit, or ameliorate the symptoms of inflammatory bowel
disease in the subject when compared to a second subject in a
control group in which the binding system was not administered. In
some embodiments, the symptom is selected from the group consisting
of a stool score, heartburn, indigestion, urgency of defecation,
nausea, vomiting, stomach pain, and bloating.
[0020] The teachings are also directed to a method of treating food
poisoning in a subject. In some embodiments, the method comprises
orally administering an effective amount of a binding system taught
herein to the subject. The binding system can prevent, inhibit, or
ameliorate the symptoms of food poisoning in the subject when
compared to a second subject in a control group in which the
binding system was not administered. In some embodiments, the
symptom is selected from the group consisting of a stool score,
heartburn, indigestion, urgency of defecation, nausea, vomiting,
stomach pain, and bloating.
[0021] The teachings are also directed to a method of treating a
wound on a tissue of a subject. In some embodiments, the method
comprises administering an effective amount of a binding system
taught herein to a wound of the subject. The binding system can
enhance the rate of healing in the subject when compared to a
second subject in a control group in which the binding system was
not administered. In some embodiments, the wound is to a dermal
tissue, mucosal tissue, or gastrointestinal tissue.
[0022] The teachings are also directed to a method of improving the
gastrointestinal health of in a subject. In some embodiments, the
method comprises orally administering a binding system taught
herein, wherein, the binding system improves the gastrointestinal
health in the subject when compared to a second subject in a
control group in which the binding system was not administered.
[0023] The teachings are also directed to a stabilized reagent pair
for aqueous transport to a target site. In some embodiments, the
reagent pair comprises a tannin having a molecular weight ranging
from about 500 Daltons to about 4000 Daltons; and, hydrogen
peroxide. The hydrogen peroxide can be hydrogen bonded to the
tannin at a tannin:peroxide weight ratio that ranges from about
1:1000 to about 10:1; the binding system can be bioactivated at a
target site having an oxidoreductase enzyme; and, the binding
molecule binds to the target site.
[0024] The teachings are also directed to a pharmaceutical
formulation comprising the a reagant pair taught herein, and a
pharmaceutically acceptable excipient. The tannin can comprise a
catechin, and the tannin:peroxide ratio can ranges from about 1:10
to about 1:50. In some embodiments, the oxidoreductase can comprise
a peroxidase; and, there can be no, or substantially no, unbound
hydrogen peroxide in the formulation.
[0025] One of skill reading the teachings that follow will
appreciate that the concepts can extend into additional embodiments
that go well-beyond a literal reading of the claims, the inventions
recited by the claims, and the terms recited in the claims.
BRIEF DESCRIPTION OF THE FIGURES
[0026] FIGS. 1A and 1B illustrate the surprising results of adding
the binding system to the drinking water of piglets, according to
some embodiments.
[0027] FIG. 2 shows the minimal inhibitory concentration (MIC)
tests for a composition of 50/50 pomegranate-green tea extract
binding system with hydrogen peroxide at a ratio of 10:1 for the
hydrogen peroxide:plant compound (molar wt/dry wt). compared to the
MIC for other common antimicrobial compounds taken from published
data, according to some embodiments.
[0028] FIG. 3 shows the binding system's the effective inhibition
of a broad spectrum of bacteria by the binding system, according to
some embodiments.
[0029] FIG. 4 shows effective reduction of virus maintaining the
host cell culture viability, according to some embodiments.
[0030] FIGS. 5A and 5B are studies showing significant elevation of
polymixin B inhibition, according to some embodiments.
[0031] FIGS. 6A and 6F show the rapid resolution of acute watery
diarrhea in 86 subjects, according to some embodiments.
DETAILED DESCRIPTION
[0032] The teachings provided herein generally relate to
site-activated binding systems that selectively increase the
bioactivity of phenolic compounds at a target site. More
particularly, the systems taught here include a phenolic compound
bound to a reactive oxygen species, wherein the phenolic compound
and the reactive oxygen species react at a target area in the
presence of an oxidoreductase enzyme to provide a site-specific
bioactivation of the binding system.
[0033] Without intending to be bound by any theory or mechanism of
action, the phenolic compounds taught herein are selected to form
multiple hydrogen bonds with a reactive oxygen species to form a
binding system that is deliverable to a target site as a stable, or
substantially stable, structure. The structure has a targeted and
enhanced effect from the selective and localized, site-activation
of the binding pair at the target site when compared to the effect
observed from administration of the phenolic compound alone. Such a
composition can be delivered to a target site, for example, in a
polar solution such as water or an alcohol. In some embodiments,
the reactive oxygen species is hydrogen peroxide, and at least a
substantial amount of the hydrogen peroxide remains bound, and thus
stable or substantially stable, with the phenolic compound.
[0034] The teachings include a binding system that selectively
increases the bioactivity of phenolic compounds at a target site.
In some embodiments, the system can include a phenolic compound
component and a reactive oxygen species component. The phenolic
compound component can comprise a tannin having a molecular weight
ranging from about 500 Daltons to about 4000 Daltons; and, the
reactive oxygen species component can comprise hydrogen peroxide.
In some embodiments, the hydrogen peroxide can be releasably bound
to the tannin at a tannin:peroxide weight ratio (a molar weight
ratio) that ranges from about 1:1000 to about 10:1. In some
embodiments, the weight ratio of the tannin:peroxide ranges from
about 1:1 to about 1:50. And, in some embodiments, the binding
system is bioactivated at a target site having an oxidoreductase
enzyme that is expressed in response to a tissue damage. In these
embodiments, the phenolic compound component can bind to the target
site selectively. Moreover, in some embodiments, the binding system
contains no, or substantially no, unbound hydrogen peroxide prior
to the bioactivating at the target site. The teachings also include
a pharmaceutical formulation comprising the binding systems taught
herein and a pharmaceutically acceptable excipient.
[0035] In some embodiments, the binding molecule comprises a
hydrolysable tannin. In some embodiments, the binding molecule
comprises a condensed tannin. And, in some embodiments, the binding
molecule comprises a combination of a hydrolysable tannin and a
condensed tannin.
[0036] In some embodiments, the phenolic compound component
comprises a flavanol. In some embodiments, the phenolic compound
component comprises a catechin. And, in some embodiments, the
phenolic compound component comprises gallic acid, epigallic acid,
or a combination thereof.
[0037] The terms "composition," "compound," "binding system," and
"binding pair," can be used interchangeably in some embodiments
and, it should be appreciated that a "formulation" can comprise a
composition, compound, binding system or binding pair presented
herein. Likewise, in some embodiments, the binding systems can also
be referred to as an "agent," a "bioactive agent," or a
"supplement" whether alone, in a pharmaceutically acceptable
composition or formulation, and whether in a liquid or dry form.
Moreover, the term "bioactivity" can refer to the increase in
function of the phenolic compound that occurs through the use of
the binding systems provided herein, where the function can refer
to an increase in the binding of the phenolic compound at a target
site upon bioactivation.
[0038] One of skill will appreciate that the term "bind,"
"binding," "bound," "attached," "connected," "chemically
connected," or "chemically attached" can be used interchangeably,
in some embodiments. Such terms can refer to any chemical bonding
mechanism known to one of skill, such as covalent, ionic,
dipole-dipole interactions, London dispersion forces, and hydrogen
bonding, for example. In some embodiments, the binding system
comprises a phenolic compound sharing hydrogen bonds with a
reactive oxygen species, such as hydrogen peroxide. In some
embodiments, the phenolic compound can comprise a polyphenol that
covalently binds to an amino acid or polyol.
[0039] In some embodiments, the term "target site" can be used to
refer to a select location, that provides, either endogeneously or
exogeneously, an oxidoreductase enzyme that can bioactivate a
binding system taught herein upon contact with the binding system.
In some embodiments, the target system can be in or on a subject.
In some embodiments, the target site can be located on or in a
plant or a non-living material. One of skill will appreciate that
the target can include any site of action in which the phenolic
compound can be site-activated by an oxidoreductase enzyme that is
available at the site. The oxidoreductase enzyme can be produced
endogeneously by a tissue at a target site, produced endogeneously
by a microbe, introduced exogenously to the target site, include
more than one enzyme, co-enzyme, catalyst, or cofactor, or a
combination thereof.
[0040] The target site can be a damaged tissue of a subject. As
such, the teachings include a method of treating a damaged dermal,
mucosal, or gastrointestinal tissue. In some embodiments, the
method includes administering an effective amount of a binding
system taught herein to the damaged tissue of the subject. In some
embodiments, the binding system functions as an antitoxin when
bioactivated at the target site of the damaged tissue and assists
in the healing of the damaged tissue by inactivating toxic
compounds at the target site.
[0041] One of skill will appreciate that the binding systems should
remain stable, or at least substantially stable, until useful or
activated, and this can relate to a shelf life, or a time between
creation of the binding pair and administration of the binding
pair, or some combination thereof. In some embodiments, the binding
pair is stable, or substantially stable, when usable as intended
within a reasonable amount of time. In some embodiments, the
binding pair should be usable within a reasonable time from the
making of the binding pair to the administration of the binding
pair and, in some embodiments, the binding pair should have a
reasonable commercial shelf life.
[0042] The binding pair can be considered as "stable" if the
binding pair loses less than 10% of it's original oxidation
potential, and this can be measured by comparing it's oxidation
potential after making the binding pair to the time of
administration, and this can include a reasonable shelf life, in
some embodiments. In some embodiments, the binding pair can be
considered as stable if the binding pair loses less than 5%, 3%,
2%, or 1% of it's original oxidation potential when comparing it's
oxidation potential after making the binding pair to the time of
administration, and this can include a reasonable shelf life, in
some embodiments.
[0043] The binding pair can be considered as "substantially stable"
if the binding system loses greater than about 10% of it's original
oxidation potential, as long as the composition can perform it's
intended use to a reasonable degree of efficacy. The loss can be
measured, as above, by measured by comparing it's oxidation
potential after making the binding pair to the time of
administration, and this can include a reasonable shelf life, in
some embodiments. In some embodiments, the binding pair can be
considered as substantially stable if a reactive oxygen species
loses greater than about 12%, about 15%, about 25%, about 35%,
about 45%, about 50%, about 60%, or even about 70% of it's original
oxidation potential. The loss may be measured by measured by
comparing it's oxidation potential after making the binding pair to
the time of administration, and this can include a reasonable shelf
life, in some embodiments.
[0044] In some embodiments, the binding pair is stable or
substantially stable, if useful for a period ranging from about 2
minutes to about 10 minutes, from about 10 minutes to about 30
minutes, from about 30 minutes to about one hour, from about one
hour to about 12 hours, from about 12 hours to about 1 day, from
about one day to about one week, from about 1 week to about 1
month, from about 1 month to about 3 months, from about 1 month to
a year, from 3 months to a year, from 3 months to 2 years, from 3
months to 3 years.
[0045] In some embodiments, the binding pair is stable, or
substantially stable for a period ranging from about 1 second to
about 2 days, from about 1 second to about 5 seconds, from about 5
seconds to about 10 seconds, from about 10 seconds to about 30
seconds, from about 30 seconds to about 1 minute, from about 1
minute to about 5 minutes, from about 5 minutes to about 15
minutes, from about 15 minutes to about 30 minutes, from about 30
minutes to about an hour, from about 1 hour to about 12 hours, from
about 12 hours to about 1 day, from about 1 day to about 2 days, or
any range therein. In some embodiments, the binding pair is stable,
or substantially stable for up to about 2 days, about 1 week, or
any range therein.
[0046] The stable structure of the binding system provides for,
over an extended period of time, an improved binding between the
phenolic compound and the target when compared to the binding of
the phenolic compound and the target in a diffuse solution. As
such, the site-activated binding systems generally increase the
bioactivity of the phenolic compounds at the target sites to a
surprising degree, which has been shown to result in a surprising
level of bioactivity and overall potency at target sites.
[0047] One of skill will appreciate that the phenolic compound in
the binding system can be any phenolic compound that functions
consistent with the teachings provided herein, and there are at
least several thousand phenolic compounds known to those of skill.
As such, the teachings provided herein can only include examples of
the general concepts rather than a comprehensive listing of all
possibilities and permutations of the systems that are enabled by
the teachings. Likewise, one of skill will appreciate that there
are numerous reactive oxygen species that can be used in the
systems taught herein, as long as the reactive oxygen species
function consistent with such teachings.
[0048] Generally speaking, phenolic compounds are those that
include a hydroxyl group bonded directly to an aromatic hydrocarbon
group. The simplest of the class is phenol (C.sub.6H.sub.5OH). One
of skill will appreciate that the entire class of phenolic
compounds is very large, and that not all of the phenolic compounds
can be used with the teachings provided herein. For example, phenol
is inoperable with the teachings provided herein, as it cannot
crosslink or polymerize with itself under the conditions in which
the binding systems are used. However, the person of skill will
also appreciate that the teachings provided herein can be used with
many compounds within the entire class of phenolic compounds.
[0049] In some embodiments, the phenolic compounds in the binding
systems (i) have phenolic hydroxyl groups that are oxidizable in
the presence of a reactive oxygen species and an oxidoreductase
enzyme, (ii) can crosslink or polymerize with other phenolic
compounds in the systems; and (iii) are soluble in a polar liquid,
such as water or an alcohol, for example, or at least moderately
soluble. And, in some embodiments, the phenolic compounds should
also be (iv) non-toxic to a subject upon administration.
[0050] In some embodiments, the phenolic compound has at least one
aryl group, or arene moiety, and at least two polar aromatic
groups, such as aromatic hydroxyl groups. In some embodiments, the
polar aromatic groups can be, for example, hydroxyl, amine, amide,
acyl, carboxy, or carbonyl. In some embodiments, the phenolic
compound has at least two aryl groups, and at least two hydroxyl
groups. In some embodiments, the phenolic compounds can be
naturally occurring, such as from a plant or other natural product.
And, in some embodiments, the phenolic compounds can be
synthetically or semi-synthetically produced. The compounds can be
simple monomers, oligomers, or polymers. The polymers can be in the
class of polyphenols or polymeric phenols, where one of skill will
understand that the general difference is typically that
polyphenols generally do not have a repeating unit, whereas
polymeric phenols do. There are exceptions, however, such that
groups of polyphenols and polymeric phenols can overlap. In most
embodiments, the phenolic compound used in the binding system can
be any phenolic compound taught herein, or any prodrugs, codrugs,
metabolites, analogs, homologues, congeners, derivatives, salts,
solvates, and combinations thereof.
[0051] In some embodiments, the phenolic compounds bind to hydrogen
peroxide to form a binding pair and, in some embodiments, the
binding pair remains stable, or substantial stable in water. In
some embodiments, the binding pair remains stable, or substantial
stable in an alcohol. And, in some embodiments, the binding pair
remains stable, or substantial stable, in a polar solvent such as,
for example, a saline solution, an an aqueous emulsion, a hydrogel,
and the like.
[0052] In some embodiments, the phenolic compounds are polyphenols
having molecular weights ranging from about 500 to about 4000
Daltons, having from about 12 to about 16 phenolic hydroxyl groups,
and having from about five to about seven aromatic rings, for every
about 1000 Daltons in molecular weight. In some embodiments, the
phenolic compounds function to precipitate alkaloids and proteins.
In some embodiments, the phenolic compounds can bind to amino
acids, peptides, oligopeptides, polyols, saccharides, or
combinations thereof. In some embodiments, the phenolic compounds
have at least from about 1 to about 20 polyhydroxylated phenolic
units and have at least moderate water solubility.
[0053] In some embodiments, the phenolic compounds are polyphenols
having molecular weights ranging from about 300 to about 4000
Daltons, having from about 2 to about 16 phenolic hydroxyl groups,
and having from about five to about seven aromatic rings, for every
about 1000 Daltons in molecular weight. In some embodiments, the
phenolic compounds function to precipitate alkaloids and proteins.
In some embodiments, the phenolic compounds can bind to amino
acids, peptides, oligopeptides, polyols, saccharides, or
combinations thereof. In some embodiments, the phenolic compounds
have at least from about 1 to about 20 polyhydroxylated phenolic
units and have at least moderate water solubility.
[0054] In some embodiments, the phenolic compounds are polyphenols
having molecular weights ranging from about 500 to about 4000
Daltons, greater than 12 to phenolic hydroxyl groups, and having
from about five to about seven aromatic rings, for every about 1000
Daltons in molecular weight. In some embodiments, the phenolic
compounds function to precipitate alkaloids and proteins. In some
embodiments, the phenolic compounds can bind to amino acids,
peptides, oligopeptides, polyols, saccharides, or combinations
thereof. In some embodiments, the phenolic compounds have at least
from about 1 to about 20 polyhydroxylated phenolic units and have
at least moderate water solubility.
[0055] The term "solubility" can refer to a concentration of a
solute in a solvent, for example, the phenolic compound in water.
The concentration can be expressed by mass, for example, mg of the
phenolic compound per kg of water at ambient temperature and
pressure. This ratio of mg/kg can be used interchangeably with ppm,
and ng/kg can be used interchangeably with ppb. In some
embodiments, the solubility of the phenolic compound can be higher
than about 500,000 ppm or less than about 1 ppm. In some
embodiments, the solubility of the phenolic compound range from
about 10 ppb to about 500,000 ppm, from about 100 ppb to about
250,000 ppm, from about 1 ppm to about 100,000 ppm, from about 10
ppm to about 50,000 ppm, from about 50 ppm to about 25,000 ppm,
from about 100 ppm to about 10,000 ppm, from about 100 ppm to about
100,000 ppm, from about 200 ppm to about 100,000 ppm, from about
250 ppm to about 50,000 ppm, from about 500 ppm to about 25,000 ppm
from about 250 ppm to about 10,000 ppm, or any range therein. In
some embodiments, the solubility can range from about 1 g/L to
about 10,000 g/L, from about 5 g/L to about 5000 g/L, from about 10
g/L to about 3000 g/L, from about 20 g/L to about 2000 g/L, from
about 50 g/L to about 1000 g/L, from about 100 g/L to about 500
g/L, or any range therein. For purposes of the teachings provided
herein, a compound can be considered to have a low solubility if
the solubility is less than about 50 g/L, a moderate solubility if
the solubility ranges from about 50 g/L to about 1000 g/L, and a
high solubility if the solubility is above about 1000 g/L. In some
embodiments, the phenolic compound can have a low solubility. In
some embodiments, the phenolic compound can have a moderate
solubility. And, in some embodiments, the phenolic compound can
have a high solubility.
[0056] One of skill will appreciate that the phenolic compounds can
still be useful at low solubilities in cases where the solubility
is too low to form a true solution. In some embodiments the
phenolic compounds can be ground into particles to form a colloidal
mixture or suspension that will function consistent with the
teachings provided herein. As such, liquid formulations include
colloids and suspensions in some embodiments. The formulations can
be a dispersed phase mixture in the form of colloidal aerosols,
colloidal emulsions, colloidal foams, colloidal dispersions, or
hydrosols. In some embodiments, the liquid formulation can include
particles having sizes ranging, for example, from about 5 nm to
about 200 nm, from about 5 nm to about 500 nm, from about 5 nm to
about 750 nm, from about 50 nm to about 1 um. In some embodiments,
the liquid formulations can be suspensions, in which the particle
size range from about 1 um to about 10 um, from about 1 um to about
7 um, from about 1 um to about 5 um, or any range therein. In some
embodiments, the liquid formulation can include particles having
sizes ranging from about 1 nm to about 10 um.
[0057] The functionality of a phenolic compound in the teachings
herein can, for at least the reason of solubility, depend on
molecular weight, alone or in addition to other factors discussed
herein such as, for example, extent of hydroxylation, presence and
location of ketone or quinine groups, and the presence of other
functional groups. In some embodiments, the molecular weights of
the phenolic compounds can range from about 110 Daltons to about
40,000 Daltons. In some embodiments, the molecular weights of the
phenolic compounds can range from about 200 Daltons to about 20,000
Daltons, from about 300 Daltons to about 30,000 Daltons, from about
400 Daltons to about 40,000 Daltons, from about 500 Daltons to
about 10,000 Daltons, from about 1000 Daltons to about 5,000
Daltons, from about 500 Daltons to about 4000 Daltons, from about
500 Daltons to about 3,000 Daltons, from about 300 Daltons to about
2,000 Daltons, from about 110 Daltons to about 30,000 Daltons, from
about 200 to about 5000 Daltons, or any range therein.
[0058] In some embodiments, the ratio of aromatic rings to
molecular weight of the phenolic compounds can range from about
five to about seven aromatic rings for every about 1000 Daltons. In
some embodiments, the ratio of aromatic rings to molecular weight
of the phenolic compounds can range from about 2 to about 10
aromatic rings for every about 1000 Daltons, from about 3 to about
9 aromatic rings for every about 1000 Daltons, from about 4 to
about 8 aromatic rings for every about 1000 Daltons, from about 5
to about 7 aromatic rings for every about 1000 Daltons, from about
1 to about 5 for every about 500 Daltons, from about 1 to about 4
for every about 500 Daltons, from about 1 to about 3 for every
about 500 Daltons, from about 2 to about 4 for every about 500
Daltons, or any range therein.
[0059] One of skill will appreciate that the phenolic compounds
should have functional groups that are capable of releasably
bonding to a reactive oxygen species, in a stable or substantially
stable form, until released upon bioactivation at a target site. In
some embodiments, a releasable bond can include any bond other than
a covalent bond. In some embodiments, a releasable bond is a
hydrogen bond. As such, the phenolic compounds should be capable of
forming, for example, a hydrogen bond with a reactive oxygen
species upon such bioactivation. In some embodiments, the phenolic
compound shares hydrogen bonding with hydrogen peroxide and is
released through a bioactivation that occurs when the binding pair
comes into contact with an oxidoreductase enzyme or other reducing
agent. In some embodiments, the phenolic compound can have
functional groups that comprise acyl, amido, amino, carbonyl,
carboxyl, hydroxyl, or peroxyl functionality. In some embodiments,
the hydrogen bond between the reactive oxygen species and the
phenolic compound can include any hydrogen donor and any hydrogen
acceptor having an available lone pair of electrons. In some
embodiments, the hydrogen acceptor can include, for example a N, O,
or F atom, or a combination thereof. In some embodiments, the
phenolic compound can have such a functionality, can be derivatized
to have such a functionality, can be linked to another compound
having such a functionality, can be placed in a carrier having such
a functionality, or some combination thereof.
[0060] In some embodiments, phenolic compounds can include simple
phenols, such as those containing 6 carbons, a C6 structure, and 1
phenolic cycle, such as the benzene alcohols, examples of which
include phenol, benzene diols and it's isomers such as catechol,
and the benzenetriols. In some embodiments, phenolic compounds can
include phenolic acids and aldehydes, such as those containing 7
carbons, a C6-C1 structure, and 1 phenolic cycle, examples of which
include gallic acid and salicylic acids. In some embodiments,
phenolic compounds can include, for example, tyrosine derivatives,
and phenylacetic acids, such as those containing 8 carbons, a C6-C2
structure, and 1 phenolic cycle, examples of which include
3-acetyl-6-methoxybenzaldehyde, tyrosol, and p-hydroxyphenylacetic
acid. In some embodiments, phenolic compounds can include
hydroxycinnamic acids, phenylpropenes, chromones, such as those
containing 9 carbons, a C6-C3 structure, and 1 phenolic cycle,
examples of which include caffeic acid, ferulic acids, myristicin,
eugenol, umbelliferone, aesculetin, bergenon, and eugenin. In some
embodiments, phenolic compounds can include naphthoquinones, such
as those containing 10 carbons, a C6-C4 structure, and 1 phenolic
cycle, examples of which include juglone and plumbagin. In some
embodiments, phenolic compounds can include xanthonoids, such as
those containing 13 carbons, a C6-C1-C6 structure, and 2 phenolic
cycles, examples of which include mangiferin. In some embodiments,
phenolic compounds can include stilbenoids, and anthraquinones,
such as those containing 14 carbons, a C6-C2-C6 structure, and 2
phenolic cycles, examples of which include resveratrol and emodin.
In some embodiments, phenolic compounds can include chalconoids,
flavonoids, isoflavonoids, and neoflavonoids, such as those
containing 15 carbons, a C6-C3-C6 structure, and 2 phenolic cycles,
examples of which include quercetin, myricetin, luteolin, cyanidin,
and genistein. In some embodiments, phenolic compounds can include
lignans and neolignans, such as those containing 18 carbons, a
C6-C3-C6 structure, and 2 phenolic cycles, examples of which
include pinoresinol and eusiderin. In some embodiments, phenolic
compounds can include biflavonoids, such as those containing 30
carbons, a (C6-C3-C6).sub.2 structure, and 4 phenolic cycles,
examples of which include amentoflavone. In some embodiments,
phenolic compounds can include polyphenols, polyphenolic proteins,
lignins, and catechol melanins, such as those containing >30
carbons. In these embodiments, the phenolic compounds can have, for
example, a (C6-C3).sub.n structure, a (C6), structure, a
(C6-C3-C6).sub.n structure, or some combination thereof, as well as
greater than about 12 phenolic cycles. Examples of such embodiments
can include, for example, the flavolans, in the class of condensed
tannins.
[0061] In some embodiments, the phenolic compounds are natural
phenols that can be enzymatically polymerized. Derivatives of
natural phenols can also be used in some embodiments. These
embodiments can include phenolic compounds having less than 12
phenolic groups, such that they can range from monophenols to
oligophenols. In some embodiments, the natural phenols are found in
plants, have an antioxidant activity, or a combination thereof.
Examples of the natural phenols include, for example, catechol- and
resorcinol-types (benzenediols) with two phenolic hydroxy groups,
and pyrogallol- and phloroglucinol-types (benzenetriols) with three
hydroxy groups. Natural phenols may have heteroatom substituents
other than hydroxyl groups, ether and ester linkages, carboxylic
acid derivatives, or some combination thereof. In some embodiments,
the natural phenols include natural phenol drugs and their
derivatives. Examples of such drugs include, but are not limited
to, anthraquinone drugs, flavone drugs, and flavonol drugs.
Examples of anthraquinone drugs include, but are not limited to,
aloe emodin, aquayamycin, and diacerein. Examples of flavone drugs
include, but are not limited to, ansoxetine and hidrosmin. Examples
of flavonol drugs include, but are not limited to, monoxerutin and
troxerutin.
[0062] In some embodiments, the phenolic compound is a tannin, a
polyphenolic phenylpropanoid, or a combination thereof. In some
embodiments, the tannin is a hydrolysable tannin, a condensed
tannin, or a combination thereof. Hydrolysable tannins can be
found, for example, in chinese gall, which is almost pure in that
it has no or substantially no condensed tannins. Condensed tannins
can be found, for example, in green tea, which is also almost pure
that it has no or substantially no hydrolysable tannins.
[0063] Examples of hydrolysable tannin can include gallotannic
acids, quercitannic acids, ellagitannins, gallotannin, pentagalloyl
glucose, galloylquinic acid, galloyl-shikimic acid, and
punicalagin. In some embodiments, the hydrolysable tannin is a
gallotannin or ellagitannin, and isomers thereof, such as isomers
that can precipitate protein. Examples of gallotannins include the
gallic acid esters of glucose in tannic acid
(C.sub.76H.sub.52O.sub.46) and pentagalloyl glucose (PGG), and
isomers thereof, such as the isomers of PGG that function to
precipitate proteins. Examples of an ellagitannin include castalin
and punicalagin. In some embodiments, the tannin is a gallic acid
ester having a molecular weight ranging from about 500 Daltons to
about 3000 Daltons. In some embodiments, the tannin is a
proanthocyanidin having a molecular weight of up to about 20,000
Daltons. In some embodiments, the hydrolysable tannins are
derivatives of gallic acid and characterized by a glucose, quinic
acid or shikimic acid core with its hydroxyl groups partially or
totally esterified with gallic acid or ellagic acid groups. The
compounds can have 3 to 12 galloyl residues but may be further
oxidatively crosslinked and complex. Hydrolysable tannins can be
readily synthesized, for example, to obtain a phenolic compound
with a high number of polar functional groups that form multiple,
stable hydrogen bonds between the tannin and hydrogen peroxide in
the binding system.
[0064] It should be appreciated that, while hydrolysable tannins
and most condensed tannins are water soluble, some very large
condensed tannins are insoluble. In some embodiments, the phenolic
compound can comprise a hydrolysable tannin such as, for example,
burkinabin C, castalagin, castalin, casuarictin, chebulagic acid,
chebulinic acid, corilagin, digallic acid, ellagitannin, gallagic
acid, gallotannin, glucogallin, grandinin, hexahydroxydiphenic
acid, pentagalloyl glucose, punicalagin alpha, punicalagins,
raspberry ellagitannin, roburin A, stenophyllanin A, stenophyllanin
A, tannate, tannic acid, tellimagrandin II, terflavin B, or
3,4,5-tri-O-galloylquinic acid.
[0065] In some embodiments, the phenolic compound can be a
flavonoid which includes several thousand natural phenol compounds.
Examples of the flavonoids include the flavonols, flavones,
flavan-3ol (catechins), flavanones, anthocyanidins, isoflavonoids,
and hybrids of any combination of these compounds. In some
embodiments, the phenolic compounds are the hydrolysable tannins
such as, for example, gallic acid. In some embodiments, the
phenolic compounds are the lignins such as, for example, cinnamic
acid. In some embodiments, the phenolic units can be dimerized or
further polymerized to form any of a variety of hybrids. For
example, ellagic acid is a dimer of gallic acid and forms the class
of ellagitannins, or a catechin and a gallocatechin can combine to
form theaflavin or the large class of thearubigins found in tea. In
another example, a flavonoid and a lignan can combine to form a
hybrid, such a flavonolignans.
[0066] In some embodiments, the phenolic compound can be a
flavan-3ol. Examples include the catechins and the catechin
gallates, where the catechin gallates are gallic acid esters of the
catechins. In some embodiments, the phenolic compound is a catechin
or epicatechin compound (the cis- or trans-isomers). In some
embodiments, the phenolic compound is (-)-epicatechin or
(+)-catechin. In some embodiments, the phenolic compound is
epigallocatechin (EGC) or gallocatechin (EC). In some embodiments,
the phenolic compound is a catechin gallate, such as
epigallocatechin gallate (EGCG)
[0067] In some embodiments, the phenolic compound can be selected
from the group of flavones consisting of apigenin, luteolin,
tangeritin, flavonols, isorhamnetin, kaempferol, myricetin (e.g.,
extractable from walnuts), proanthocyanidins or condensed tannins,
and quercetin and related phenolic compounds, such as rutin.
[0068] In some embodiments, the phenolic compound can be selected
from the group of flavanones consisting of eriodictyol, hesperetin
(metabolizes to hesperidin), and naringenin (metabolized from
naringin).
[0069] In some embodiments, the phenolic compound can be selected
from the group of flavanols consisting of catechin, gallocatechin
and their corresponding gallate esters, epicatechin,
epigallocatechin and their corresponding gallate esters, theaflavin
and its gallate esters, thearubigins, isoflavone phytoestrogens
(found primarily in soy, peanuts, and other members of the Fabaceae
family), daidzein, genistein, glycitein, stilbenoids, resveratrol
(found in the skins of dark-colored grapes, and concentrated in red
wine), pterostilbene (methoxylated analogue of resveratrol,
abundant in Vaccinium berries), anthocyanins, cyanidin,
delphinidin, malvidin, pelargonidin, peonidin, and petunidin. And,
In some embodiments, the phenolic compound can be ubiquinol an
electron-rich (reduced) form of coenzyme Q10.
[0070] In some embodiments, the phenolic compound can be selected
from the group of carotenoid terpenoid consisting of
alpha-carotene, astaxanthin (found naturally in red algae and
animals higher in the marine food chain, a red pigment familiarly
recognized in crustacean shells and salmon flesh/roe),
beta-carotene (found in high concentrations in butternut squash,
carrots, orange bell peppers, pumpkins, and sweet potatoes),
canthaxanthin, lutein (found in high concentration in spinach,
kiwifruit and red peppers), lycopene (found in high concentration
in ripe red tomatoes and watermelons) and zeaxanthin (the main
pigment found in yellow corn, also abundant in kiwifruit).
[0071] In some embodiments, the phenolic compound can be selected
from the group of phenolic acids and their esters consisting of
chicoric acid (another caffeic acid derivative, is found only in
the medicinal herb echinacea purpurea), chlorogenic acid (found in
high concentration in coffee (more concentrated in robusta than
arabica beans, blueberries and tomatoes, and produced from
esterification of caffeic acid), cinnamic acid and its derivatives,
such as ferulic acid (found in seeds of plants such as in brown
rice, whole wheat and oats, as well as in coffee, apple, artichoke,
peanut, orange and pineapple), ellagic acid (found in high
concentration in raspberry and strawberry, and in ester form in red
wine tannins), ellagitannins (hydrolysable tannin polymer formed
when ellagic acid, a polyphenol monomer, esterifies and binds with
the hydroxyl group of a polyol carbohydrate such as glucose),
gallic acid (found in gallnuts, sumac, witch hazel, tea leaves, oak
bark, and many other plants), gallotannins (hydrolysable tannin
polymer formed when gallic acid, a polyphenol monomer, esterifies
and binds with the hydroxyl group of a polyol carbohydrate such as
glucose), rosmarinic acid (found in high concentration in rosemary,
oregano, lemon balm, sage, and marjoram), and salicylic acid (found
in most vegetables, fruits, and herbs; but most abundantly in the
bark of willow trees, from where it was extracted for use in the
early manufacture of aspirin).
[0072] In some embodiments, the phenolic compound can be selected
from the group of non-flavonoid phenolics consisting of curcumin
(has low bioavailability, because, much of it is excreted through
glucuronidation, but bioavailability can be substantially enhanced
by solubilization in a lipid (oil or lecithin), heat, addition of
piperine, or through nanoparticularization, flavonolignans, for
example, silymarin which is a mixture of flavonolignans extracted
from milk thistle), eugenol and xanthones (mangosteen, for example,
is purported to contain a large variety of xanthones, some of
which, like mangostin are believed to be present only in the
inedible shell).
[0073] In some embodiments, the phenolic compound can have a low
molecular weight (less than about 400 Daltons), selected from the
group consisting of caffeic acid, gentisic acid, protocatechuic
acid, phenylacetic acid, gallic acid, phloroglucinol carboxylic
acid, and derivatives thereof. Such compounds can form a
sufficiently soluble binding pair, and their relatively high
hydroxyl group to molecular weight ratio creates favorable
conditions for obtaining the intermolecular hydrogen bonds desired
for the binding systems.
[0074] In some embodiments, the phenolic compounds can be from a
natural extract, such as an extract of a plant or other natural
product. See, for example, U.S. Published Patent Application Nos.
20100158885 and 20110070198, each of which is hereby incorporated
by reference herein in its entirety. Those skilled in the art of
such extracts will understand that extracts of plant materials are
not typically pure in one type of phenolic compound. Plant tannin
extracts, for example, typically comprise heterogeneous mixtures
and derivatives of the above classes.
[0075] In some embodiments, the phenolic compound is extracted from
a whole or partial plant tissue selected from the group consisting
of seeds and fruits; ovaries; juice; pulp; galls; husks; bark;
stems; leaves; flowers; sheaths; hulls; sprouts; bulbs; hips;
tubers; roots of grains; grasses; legumes; trees; vegetables;
medicinal herbs; tea leaves; algaes; marine plants; and, forages.
One of skill will appreciate that the type and content of phenolic
compound obtained can be expected to vary with the species, season,
geographical location, cultivation, and storage.
[0076] Generally speaking, the reactive oxygen species include
those that can facilitate the oxidation of a phenol hydroxyl group
to a ketone group and form a reactive quinone structure upon the
bioactivation. In some embodiments, the reactive oxygen species can
include hydrogen peroxide, superoxide anion, singlet oxygen, or a
hydroxyl radical. In some embodiments, the reactive oxygen species
is hydrogen peroxide. In some embodiments, the reactive oxygen
species is hydrogen peroxide.
[0077] In some embodiments, the reactive oxygen species is hydrogen
peroxide or a material that release hydrogen peroxide including,
but not limited to, hydration of adducts of hydrogen peroxide such
as carbamide peroxide, magnesium peroxide, and sodium percarbonate;
amino perhydrates; superoxide dismutase decomposition of ozone,
superoxides or superoxide salts; glucose oxidase and glucose,
aqueous dilution of honey; H.sub.2O.sub.2 production by
lactobacillus; catalytic quinone hydrogenation; superoxides; and,
superoxide dismutase. In some embodiments, the reactive oxygen
species can include peroxide ion, organic peroxides, organic
hydroperoxides, peracid superoxides, dioxygenyls, ozones, and
ozonides.
[0078] And, generally speaking, one of skill will appreciate that
there are a wide variety of enzymes that can activate the binding
system taught herein. And, the enzyme that bioactivates the binding
system is, at least in part, responsible for the selectivity of the
binding systems at a target site. Generally, the enzymes fall into
the classes of oxidoreductases. As such, there are several enzymes
and isozymes that will be present at a target site and capable of
bioactivating the binding systems. In some embodiments, the
oxidoreductases can be categorized into about 22 classes, and the
selectivity of the bioactivation of the binding system at a target
site depends, at least in part, on the selectivity of the
oxidoreductase at the target site. In some embodiments, the
oxidoreductase can include those oxidoreductases that act on the
CH--OH group of donors (alcohol oxidoreductases, for example; EC
Number class 1.1). In some embodiments, the oxidoreductase can
include those oxidoreductases that act on diphenols and related
substances as donors (catechol oxidase, for example, EC Number
class 1.10). In some embodiments, the oxidoreductase can include
those oxidoreductases that act on peroxide as an acceptor
(peroxidases, such as horseradish peroxidase and catalase; EC
Number class 1.11). In some embodiments, the oxidoreductase can
include those oxidoreductases that act on phenols as an acceptor
(tyrosinases, for example; EC Number class 1.14). Examples of other
useful enzymes for the teachings provided herein include, but are
not limited to, glutathione peroxidase 1 and 4 (in many mammalian
tissues), glutathione peroxidase 2 (in intestinal and extracellular
mammalian tissues), glutathione peroxidase 3 (in plasma mammalian
tissues), lactoperoxidase, myeloperoxidase (in salivary &
mucosal mammalian tissues), myeloperoxidase (in neutrophil
mammalian tissues), cytochrome peroxidase (in yeasts such as
Candida albicans) and horseradish peroxidase (common to show in
vitro activity). One of skill will appreciate that oxidoreductases
are selective and, in some embodiments, the oxidoreductase can
include an alternate enzyme that are selective for a binding system
having a phenolic compound that acts as a substrate for the
alternative enzyme.
[0079] In some embodiments, the oxidoreductases include
mono-oxygenases such as, for example, phenylalaning monooxygenase,
tyrosine monooxygenase, and tryptophan monooxygenase. In some
embodiments, the oxidoreductases include dioxygenases such as, for
example, tryptophan dioxygenase, homogentisate dioxygenase,
trimethyl lysine dioxygenase, and nitric oxide synthase. In some
embodiments, the oxidoreductases include peroxidases such as, for
example, catalase, myeloperoxidase, thyroperoxidase. In some
embodiments, the oxidoreductases act in the presence of a co-factor
or co-enzyme, such as nicotinamide adenine dinucleotide phosphate
(NADP) or nicotinamide adenine dinucleotide (NAD).
[0080] The compounds described herein can have one or more chemical
substitutions. In some embodiments, the substitution can be at any
location on the molecule or macromolecule and may be designated as
an "R-group." The R groups can be used to represent nearly any
chemical moiety, or functional group. For example, one of skill
would or could substitute the group and still obtain the functions
consistent with the teachings provided herein. For example, in some
embodiments, an R group can be an alkyl, alkanyl, alkenyl, alkynyl,
alkoxy, acyl, aryl, aralkyl, halo, heteroalkyl, heteroalkanyl,
heteroalkenyl, heteroalkynyl, heteroaryl, heteroaralkyl, and the
like.
[0081] "Alkyl," by itself or as part of another substituent, can
refer to a saturated or unsaturated, branched, straight-chain or
cyclic monovalent hydrocarbon radical derived by the removal of one
hydrogen atom from a single carbon atom of a parent alkane, alkene
or alkyne. Typical alkyl groups can include, but are not limited
to, methyl; ethyls such as ethanyl, ethenyl, ethynyl; propyls such
as propan-1-yl, propan-2-yl, cyclopropan-1-yl, prop-1-en-1-yl,
prop-1-en-2-yl, prop-2-en-1-yl (allyl), cycloprop-1-en-1-yl;
cycloprop-2-en-1-yl, prop-1-yn-1-yl, prop-2-yn-1-yl, etc.; butyls
such as butan-1-yl, butan-2-yl, 2-methyl-propan-1-yl,
2-methyl-propan-2-yl, cyclobutan-1-yl, but-1-en-1-yl,
but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl,
but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl,
cyclobut-1-en-1-yl, cyclobut-1-en-3-yl, cyclobuta-1,3-dien-1-yl,
but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.
The term "alkyl" is specifically intended to include groups having
any degree or level of saturation, i.e., groups having exclusively
single carbon-carbon bonds, groups having one or more double
carbon-carbon bonds, groups having one or more triple carbon-carbon
bonds and groups having mixtures of single, double and triple
carbon-carbon bonds. Where a specific level of saturation is
intended, the expressions "alkanyl," "alkenyl," and "alkynyl" are
used. In some embodiments, an alkyl group comprises from 1 to 20
carbon atoms (C.sub.1-C.sub.20 alkyl). In some embodiments, an
alkyl group comprises from 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6,
1 to 7, 1 to 8, 1 to 9, or 1 to 10 carbon atoms (C.sub.1-C.sub.10
alkyl). In some embodiments, an alkyl group comprises from about 1
to 3 to about 1 to 6 carbon atoms (from C.sub.1-C.sub.3 to
C.sub.1-C.sub.6 alkyl). In some embodiments, an alkyl group
comprises from 1 to 4 carbon atoms (C.sub.1-C.sub.4 alkyl).
[0082] "Alkanyl," by itself or as part of another substituent, can
refer to a saturated branched, straight-chain or cyclic alkyl
radical derived by the removal of one hydrogen atom from a single
carbon atom of a parent alkane. Typical alkanyl groups can include,
but are not limited to, methanyl; ethanyl; propanyls such as
propan-1-yl, propan-2-yl (isopropyl), cyclopropan-1-yl, etc.;
butanyls such as butan-1-yl, butan-2-yl (sec-butyl),
2-methyl-propan-1-yl (isobutyl), 2-methyl-propan-2-yl (t-butyl),
cyclobutan-1-yl, etc.; and the like.
[0083] "Alkenyl," by itself or as part of another substituent, can
refer to an unsaturated branched, straight-chain or cyclic alkyl
radical having at least one carbon-carbon double bond derived by
the removal of one hydrogen atom from a single carbon atom of a
parent alkene. The group may be in either the cis or trans
conformation about the double bond(s). Typical alkenyl groups can
include, but are not limited to, ethenyl; propenyls such as
prop-1-en-1-yl, prop-1-en-2-yl, prop-2-en-1-yl (allyl),
prop-2-en-2-yl, cycloprop-1-en-1-yl; cycloprop-2-en-1-yl; butenyls
such as but-1-en-1-yl, but-1-en-2-yl, 2-methyl-prop-1-en-1-yl,
but-2-en-1-yl, but-2-en-1-yl, but-2-en-2-yl, buta-1,3-dien-1-yl,
buta-1,3-dien-2-yl, cyclobut-1-en-1-yl, cyclobut-1-en-3-yl,
cyclobuta-1,3-dien-1-yl, etc.; and the like.
[0084] "Alkynyl," by itself or as part of another substituent can
refer to an unsaturated branched, straight-chain or cyclic alkyl
radical having at least one carbon-carbon triple bond derived by
the removal of one hydrogen atom from a single carbon atom of a
parent alkyne. Typical alkynyl groups can include, but are not
limited to, ethynyl; propynyls such as prop-1-yn-1-yl,
prop-2-yn-1-yl, etc.; butynyls such as but-1-yn-1-yl,
but-1-yn-3-yl, but-3-yn-1-yl, etc.; and the like.
[0085] "Alkoxy," by itself or as part of another substituent, can
refer to a radical of the formula --O--R.sup.400, where R.sup.400
is alkyl or substituted alkyl as defined herein.
[0086] "Acyl" by itself or as part of another substituent can refer
to a radical --C(O)R.sup.401, where R.sup.401 is hydrogen, alkyl,
substituted alkyl, aryl, substituted aryl, arylalkyl, substituted
arylalkyl, heteroalkyl, substituted heteroalkyl, heteroarylalkyl or
substituted heteroarylalkyl as defined herein. Representative
examples include, but are not limited to formyl, acetyl,
cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl,
benzylcarbonyl and the like.
[0087] "Aryl," by itself or as part of another substituent, can
refer to a monovalent aromatic hydrocarbon group derived by the
removal of one hydrogen atom from a single carbon atom of a parent
aromatic ring system, as defined herein. Typical aryl groups can
include, but are not limited to, groups derived from aceanthrylene,
acenaphthylene, acephenanthrylene, anthracene, azulene, benzene,
chrysene, coronene, fluoranthene, fluorene, hexacene, hexaphene,
hexylene, as-indacene, s-indacene, indane, indene, naphthalene,
octacene, octaphene, octalene, ovalene, penta-2,4-diene, pentacene,
pentalene, pentaphene, perylene, phenalene, phenanthrene, picene,
pleiadene, pyrene, pyranthrene, rubicene, triphenylene,
trinaphthalene and the like. In some embodiments, an aryl group
comprises from 6 to 20 carbon atoms (C.sub.6-C.sub.20 aryl). In
some embodiments, an aryl group comprises from 6 to 15 carbon atoms
(C.sub.6-C.sub.15 aryl). In still other embodiments, an aryl group
comprises from 6 to 15 carbon atoms (C.sub.6-C.sub.10 aryl). In
some embodiments, an aryl group can be an arene moiety that forms
at least a part of a molecule used in the teachings herein.
[0088] "Arylalkyl," by itself or as part of another substituent,
can refer to an acyclic alkyl group in which one of the hydrogen
atoms bonded to a carbon atom, typically a terminal or sp.sup.3
carbon atom, is replaced with an aryl group as, as defined herein.
Typical arylalkyl groups can include, but are not limited to,
benzyl, 2-phenylethan-1-yl, 2-phenylethen-1-yl, naphthylmethyl,
2-naphthylethan-1-yl, 2-naphthylethen-1-yl, naphthobenzyl,
2-naphthophenylethan-1-yl and the like. Where specific alkyl
moieties are intended, the nomenclature arylalkanyl, arylalkenyl
and/or arylalkynyl is used. In some embodiments, an arylalkyl group
is (C.sub.6-C.sub.30) arylalkyl, e.g., the alkanyl, alkenyl or
alkynyl moiety of the arylalkyl group is (C.sub.1-C.sub.10) alkyl
and the aryl moiety is (C.sub.6-C.sub.20) aryl. In some
embodiments, an arylalkyl group is (C.sub.6-C.sub.20) arylalkyl,
e.g., the alkanyl, alkenyl or alkynyl moiety of the arylalkyl group
is (C.sub.1-C.sub.8) alkyl and the aryl moiety is
(C.sub.6-C.sub.12) aryl. In still other embodiments, an arylalkyl
group is (C.sub.6-C.sub.15) arylalkyl, e.g., the alkanyl, alkenyl
or alkynyl moiety of the arylalkyl group is (C.sub.1-C.sub.5) alkyl
and the aryl moiety is (C.sub.6-C.sub.10) aryl.
[0089] "Compounds" can refer to compounds encompassed by structural
formulae disclosed herein and includes any specific compounds
within these formulae whose structure is disclosed herein.
Compounds may be identified either by their chemical structure
and/or chemical name. When the chemical structure and chemical 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 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 depicted herein encompass all possible tautomeric forms
of the illustrated compounds. The compounds described also include
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 or unhydrated 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. In
general, all physical forms are equivalent for the uses
contemplated herein and are intended to be within the scope of the
present invention. Further, it should be understood, when partial
structures of the compounds are illustrated, that brackets indicate
the point of attachment of the partial structure to the rest of the
molecule.
[0090] In some embodiments, the compounds can have one or more
electron withdrawing group. An "electron withdrawing group" can
refer to a chemical functional group that draws electrons away from
a reaction center. Examples of electron withdrawing groups can
include halogens (e.g., Cl,). nitriles (e.g., CN); carbonyls (e.g.,
CO), and nitro groups (NO.sub.2). Any one or any combination of
nitro, acyl, formyl, alkylsulfonyl, arylsulfonyl, trifluoromethyl,
cyano, halo (e.g., fluoro, chloro, bromo, and iodo) moieties, and
other electron-withdrawing groups can be used in some embodiments.
In some embodiments, halo, nitrate and fluoromethyl groups
(CF.sub.3, CHF.sub.2 or CH.sub.2F) can be suitable electron
withdrawing groups. One of skill will appreciate that there are
several atoms, chemical groups, or structures, i.e., chemical
moieties, that can function as an electron withdrawing group for
purposes of the teachings provided herein. Whether a particular
chemical moiety acts as an electron withdrawing group can depend on
the nature of the neighboring chemical moiety or moieties, as an
electron withdrawing group draws electron density from neighboring
atoms towards itself, usually by resonance or inductive effects. In
some embodiments, a weaker base can draw electrons from stronger
base. For purposes of illustration, trifluoroacetate ion is a
weaker base than acetate ion because the trifluoromethyl group is
able to draw electron density away from the carboxylate when in a
neighboring chemical relationship, making the trifluoromethyl group
an electron withdrawing group in this situation. One of skill will
appreciate that electron withdrawing groups can be added in one or
more positions of a chemical structure to produce a cumulative
effect, and each electron withdrawing group can be independently
selected.
[0091] "Halogen", or "halo," by itself or as part of another
substituent can refer to a radical --F, --Cl, --Br or --I.
[0092] "Heteroalkyl," "Heteroalkanyl," "Heteroalkenyl" and
"Heteroalkynyl," by themselves or as part of other substituents,
refer to alkyl, alkanyl, alkenyl and alkynyl groups, respectively,
in which one or more of the carbon atoms (and optionally any
associated hydrogen atoms), are each, independently of one another,
replaced with the same or different heteroatoms or heteroatomic
groups. Typical heteroatoms or heteroatomic groups which can
replace the carbon atoms include, but are not limited to, --O--,
--S--, --N--, --Si--, --NH--, --S(O)--, --S(O).sub.2--, --S(O)NH--,
--S(O).sub.2NH-- and the like and combinations thereof. The
heteroatoms or heteroatomic groups may be placed at any interior
position of the alkyl, alkenyl or alkynyl groups. Typical
heteroatomic groups which can be included in these groups can
include, but are not limited to, --O--, --S--, --O--O--, --S--S--,
--O--S--, --NR.sup.501R.sup.502--, .dbd.N--N.dbd., --N.dbd.N--,
--N.dbd.N--NR.sup.503R.sup.404, --PR.sup.505--, --P(O).sub.2--,
--POR.sup.506--, --O--P(O).sub.2--, --SO--, --SO.sub.2--,
--SnR.sup.507R.sup.508-- and the like, where R.sup.501, R.sup.502,
R.sup.503, R.sup.504, R.sup.505, R.sup.506, R.sup.507 and R.sup.508
are independently hydrogen, alkyl, substituted alkyl, aryl,
substituted aryl, arylalkyl, substituted arylalkyl, cycloalkyl,
substituted cycloalkyl, cycloheteroalkyl, substituted
cycloheteroalkyl, heteroalkyl, substituted heteroalkyl, heteroaryl,
substituted heteroaryl, heteroarylalkyl or substituted
heteroarylalkyl.
[0093] "Heteroaryl," by itself or as part of another substituent,
can refer to a monovalent heteroaromatic radical derived by the
removal of one hydrogen atom from a single atom of a parent
heteroaromatic ring systems, as defined herein. Typical heteroaryl
groups can include, but are not limited to, groups derived from
acridine, .beta.-carboline, chromane, chromene, cinnoline, furan,
imidazole, indazole, indole, indoline, indolizine, isobenzofuran,
isochromene, isoindole, isoindoline, isoquinoline, isothiazole,
isoxazole, naphthyridine, oxadiazole, oxazole, perimidine,
phenanthridine, phenanthroline, phenazine, phthalazine, pteridine,
purine, pyran, pyrazine, pyrazole, pyridazine, pyridine,
pyrimidine, pyrrole, pyrrolizine, quinazoline, quinoline,
quinolizine, quinoxaline, tetrazole, thiadiazole, thiazole,
thiophene, triazole, xanthene, and the like. In some embodiments,
the heteroaryl group comprises from 5 to 20 ring atoms (5-20
membered heteroaryl). In some embodiments, the heteroaryl group
comprises from 5 to 10 ring atoms (5-10 membered heteroaryl).
Exemplary heteroaryl groups can include those derived from furan,
thiophene, pyrrole, benzothiophene, benzofuran, benzimidazole,
indole, pyridine, pyrazole, quinoline, imidazole, oxazole,
isoxazole and pyrazine.
[0094] "Heteroarylalkyl" by itself or as part of another
substituent can refer to an acyclic alkyl group in which one of the
hydrogen atoms bonded to a carbon atom, typically a terminal or
sp.sup.3 carbon atom, is replaced with a heteroaryl group. Where
specific alkyl moieties are intended, the nomenclature
heteroarylalkanyl, heteroarylakenyl and/or heteroarylalkynyl is
used. In some embodiments, the heteroarylalkyl group is a 6-21
membered heteroarylalkyl, e.g., the alkanyl, alkenyl or alkynyl
moiety of the heteroarylalkyl is (C.sub.1-C.sub.6) alkyl and the
heteroaryl moiety is a 5-15-membered heteroaryl. In some
embodiments, the heteroarylalkyl is a 6-13 membered
heteroarylalkyl, e.g., the alkanyl, alkenyl or alkynyl moiety is
(C.sub.1-C.sub.3) alkyl and the heteroaryl moiety is a 5-10
membered heteroaryl.
[0095] "Parent Aromatic Ring System" can refer to an unsaturated
cyclic or polycyclic ring system having a conjugated .pi. electron
system. Specifically included within the definition of "parent
aromatic ring system" are fused ring systems in which one or more
of the rings are aromatic and one or more of the rings are
saturated or unsaturated, such as, for example, fluorene, indane,
indene, phenalene, etc. Typical parent aromatic ring systems
include, but are not limited to, aceanthrylene, acenaphthylene,
acephenanthrylene, anthracene, azulene, benzene, chrysene,
coronene, fluoranthene, fluorene, hexacene, hexaphene, hexylene,
as-indacene, s-indacene, indane, indene, naphthalene, octacene,
octaphene, octalene, ovalene, penta-2,4-diene, pentacene,
pentalene, pentaphene, perylene, phenalene, phenanthrene, picene,
pleiadene, pyrene, pyranthrene, rubicene, triphenylene,
trinaphthalene and the like.
[0096] "Parent Heteroaromatic Ring System" can refer to a parent
aromatic ring system in which one or more carbon atoms (and
optionally any associated hydrogen atoms) are each independently
replaced with the same or different heteroatom. Typical heteroatoms
to replace the carbon atoms include, but are not limited to, N, P,
O, S, Si, etc. Specifically included within the definition of
"parent heteroaromatic ring system" are fused ring systems in which
one or more of the rings are aromatic and one or more of the rings
are saturated or unsaturated, such as, for example, benzodioxan,
benzofuran, chromane, chromene, indole, indoline, xanthene, etc.
Typical parent heteroaromatic ring systems include, but are not
limited to, arsindole, carbazole, .beta.-carboline, chromane,
chromene, cinnoline, furan, imidazole, indazole, indole, indoline,
indolizine, isobenzofuran, isochromene, isoindole, isoindoline,
isoquinoline, isothiazole, isoxazole, naphthyridine, oxadiazole,
oxazole, perimidine, phenanthridine, phenanthroline, phenazine,
phthalazine, pteridine, purine, pyran, pyrazine, pyrazole,
pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine,
quinazoline, quinoline, quinolizine, quinoxaline, tetrazole,
thiadiazole, thiazole, thiophene, triazole, xanthene and the
like.
[0097] "Salt" can refer to a salt of a compound, which possesses
the desired pharmacological activity of the parent compound. Such
salts include: (1) acid addition salts, formed with inorganic acids
such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric
acid, phosphoric acid, and the like; or formed with organic acids
such as acetic acid, propionic acid, hexanoic acid,
cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic
acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric
acid, tartaric acid, citric acid, benzoic acid,
3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid,
methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic
acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,
4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,
4-toluenesulfonic acid, camphorsulfonic acid,
4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic
acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary
butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic
acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic
acid, and the like; or (2) salts formed when an acidic proton
present in the parent compound is replaced by a metal ion, e.g., an
alkali metal ion, an alkaline earth ion, or an aluminum ion; or
coordinates with an organic base such as ethanolamine,
diethanolamine, triethanolamine, N-methylglucamine and the
like.
[0098] "Substituted," when used to modify a specified group or
radical, means that one or more hydrogen atoms of the specified
group or radical are each, independently of one another, replaced
with the same or different substituent(s). Substituent groups
useful for substituting saturated carbon atoms in the specified
group or radical include, but are not limited to --R.sup.a, halo,
--O.sup.-, .dbd.O, --OR.sup.b, --SR.sup.b, --S.sup.-, .dbd.S,
--NR.sup.cR.sup.c, .dbd.NR.sup.b, .dbd.N--OR.sup.b, trihalomethyl,
--CF.sub.3, --CN, --OCN, --SCN, --NO, --NO.sub.2, .dbd.N.sub.2,
--N.sub.3, --S(O).sub.2R.sup.b, --S(O).sub.2NR.sup.b,
--S(O).sub.2O.sup.-, --S(O).sub.2OR.sup.b, --OS(O).sub.2R.sup.b,
--OS(O).sub.2O.sup.-, --OS(O).sub.2OR.sup.b, --P(O)(O.sup.-).sub.2,
--P(O)(OR.sup.b)(O.sup.-), --P(O)(OR.sup.b)(OR.sup.b),
--C(O)R.sup.b, --C(S)R.sup.b, --C(NR.sup.b)R.sup.b, --C(O)O.sup.-,
--C(O)OR.sup.b, --C(S)OR.sup.b, --C(O)NR.sup.cR.sup.c,
--C(NR.sup.b)NR.sup.cR.sup.c, --OC(O)R.sup.b, --OC(S)R.sup.b,
--OC(O)O.sup.-, --OC(O)OR.sup.b, --OC(S)OR.sup.b,
--NR.sup.bC(O)R.sup.b, --NR.sup.bC(S)R.sup.b,
--NR.sup.bC(O)O.sup.-, --NR.sup.bC(O)OR.sup.b,
--NR.sup.bC(S)OR.sup.b, --NR.sup.bC(O)NR.sup.cR.sup.c,
--NR.sup.bC(NR.sup.b)R.sup.b and
--NR.sup.bC(NR.sup.b)NR.sup.cR.sup.c, where R.sup.a is selected
from the group consisting of alkyl, cycloalkyl, heteroalkyl,
cycloheteroalkyl, aryl, arylalkyl, heteroaryl and heteroarylalkyl;
each R.sup.b is independently hydrogen or R.sup.a; and each R.sup.c
is independently R.sup.b or alternatively, the two R.sup.cs are
taken together with the nitrogen atom to which they are bonded form
a 4-, 5-, 6- or 7-membered cycloheteroalkyl which may optionally
include from 1 to 4 of the same or different additional heteroatoms
selected from the group consisting of O, N and S. As specific
examples, --NR.sup.cR.sup.c is meant to include --NH.sub.2,
--NH-alkyl, N-pyrrolidinyl and N-morpholinyl.
[0099] Similarly, substituent groups useful for substituting
unsaturated carbon atoms in the specified group or radical include,
but are not limited to, --R.sup.a, halo, --O.sup.-, --OR.sup.b,
--SR.sup.b, --S--, --NR.sup.cR.sup.c, trihalomethyl, --CF.sub.3,
--CN, --OCN, --SCN, --NO, --NO.sub.2, --N.sub.3,
--S(O).sub.2R.sup.b, --S(O).sub.2O.sup.-, --S(O).sub.2OR.sup.b,
--OS(O).sub.2R.sup.b, --OS(O).sub.2O.sup.-, --OS(O).sub.2OR.sup.b,
--P(O)(O.sup.-).sub.2, --P(O)(OR.sup.b)(O.sup.-),
--P(O)(OR.sup.b)(OR.sup.b), --C(O)R.sup.b, --C(S)R.sup.b,
--C(NR.sup.b)R.sup.b, --C(O)O.sup.-, --C(O)OR.sup.b,
--C(S)OR.sup.b, --C(O)NR.sup.cR.sup.c,
--C(NR.sup.b)NR.sup.cR.sup.c, --OC(O)R.sup.b, --OC(S)R.sup.b,
--OC(O)O.sup.-, --OC(O)OR.sup.b, --OC(S)OR.sup.b,
--NR.sup.bC(O)R.sup.b, --NR.sup.bC(S)R.sup.b,
--NR.sup.bC(O)O.sup.-, --NR.sup.bC(O)OR.sup.b,
--NR.sup.bC(S)OR.sup.b, --NR.sup.bC(O)NR.sup.cR.sup.c,
--NR.sup.bC(NR.sup.b)R.sup.b and
--NR.sup.bC(NR.sup.b)NR.sup.cR.sup.c, where R.sup.a, R.sup.b and
R.sup.c are as previously defined.
[0100] Substituent groups useful for substituting nitrogen atoms in
heteroalkyl and cycloheteroalkyl groups can include, but are not
limited to, --R.sup.a, --O.sup.-, --OR.sup.b, --SR.sup.b,
--S.sup.-, --NR.sup.cR.sup.c, trihalomethyl, --CF.sub.3, --CN,
--NO, --NO.sub.2, --S(O).sub.2R.sup.b, --S(O).sub.2O.sup.-,
--S(O).sub.2OR.sup.b, --OS(O).sub.2R.sup.b, --OS(O).sub.2O.sup.-,
--OS(O).sub.2OR.sup.b, --P(O)(O.sup.-).sub.2,
--P(O)(OR.sup.b)(O.sup.-), --P(O)(OR.sup.b)(OR.sup.b),
--C(O)R.sup.b, --C(S)R.sup.b, --C(NR.sup.b)R.sup.b, --C(O)OR.sup.b,
--C(S)OR.sup.b, --C(O)NR.sup.cR.sup.c,
--C(NR.sup.b)NR.sup.cR.sup.c, --OC(O)R.sup.b, --OC(S)R.sup.b,
--OC(O)OR.sup.b, --OC(S)OR.sup.b, --NR.sup.bC(O)R.sup.b,
--NR.sup.bC(S)R.sup.b, --NR.sup.bC(O)OR.sup.b,
--NR.sup.bC(S)OR.sup.b, --NR.sup.bC(O)NR.sup.cR.sup.c,
--NR.sup.bC(NR.sup.b)R.sup.b and
--NR.sup.bC(NR.sup.b)NR.sup.cR.sup.c, where R.sup.a, R.sup.b and
R.sup.c are as previously defined.
[0101] Substituent groups from the above lists useful for
substituting other specified groups or atoms will be apparent to
those of skill in the art. The substituents used to substitute a
specified group can be further substituted, typically with one or
more of the same or different groups selected from the various
groups specified above.
Methods of Making the Binding Systems
[0102] The design of the binding systems include (i) selecting the
phenolic compound, (ii) selecting the reactive oxygen species,
(iii) selecting the ratio of phenolic compound to reactive oxygen
species, and (iv) selecting a carrier. In some embodiments, the
phenolic compound can be derivatized or attached to another
chemical moiety via a linker, or another known method such as, for
example, esterification to facilitate or improve an association
between the phenolic compound and the reactive oxygen species, as
well as to potentially modify, solubility, tissue absorption, or
toxicity.
[0103] One of skill will appreciate that, at least from the
teachings provided herein, there are a vast number of binding
systems that can be selected for bioactivation at a given target
site, the selection of which is, at least in part, dependent on
type of enzyme, co-enzymes, cofactors or catalysts present at the
target site for the bioactivation of the binding system. The design
of the binding system can include for example, (i) identifying the
target site; (ii) identifying an enzyme, co-enzymes, cofactors, or
catalysts present at the target site but not present at tissue
surrounding the target site; (iii) selecting a binding pair for
activation at the target site by the enzyme, co-enzymes, cofactors,
or catalysts; and, (iv) selecting a carrier in which the binding
pair is stable or substantially stable.
[0104] Identifying the target site includes, for example, select a
target tissue for treatment, such as a damaged tissue at which the
enzyme, co-enzymes, cofactors or catalysts present. In some
embodiments, the target site is a damaged mucosal tissue, such as a
damaged GI tissue, at which peroxidase or oxidase may be
present.
[0105] Identifying an enzyme, co-enzymes, cofactors, or catalysts
present at the target site but not present at tissue surrounding
the target site can include identifying the tissue type, and the
type of damage, as well as the presence of a microbe, for example.
Anaerobic pathogens such as Pseudomonas and Vibrio can express a
peroxide or an oxidase, making these enzymes available at the
target site.
[0106] Given the teachings provided herein, one of skill can the
select a binding pair for activation at the target site by the
enzyme, co-enzymes, cofactors, or catalysts. After the binding pair
and environment of use are known, one of skill can a carrier in
which the binding pair is stable or substantially stable. In one
example, the binding system can comprise a mixture of phenolic
compounds in a desired ratio with hydrogen peroxide. The phenolic
compounds include a mixture of a pomegranate extract and a green
tea extract, and the ratio of phenolic compound to hydrogen
peroxide can range from about 1:2 to about 1:20 on a wt/wt basis
(molar weight). The hydrogen peroxide can be added to the phenolic
compound using a concentration of about 0.1% to about 10% hydrogen
peroxide solution. One of skill can easily selected the dose for a
particular use, which will vary according to use, due to
environmental conditions at the site of use. In another example,
the binding system can comprise a mixture of phenolic compounds in
a desired ratio with hydrogen peroxide. The phenolic compounds
include a mixture of a pomegranate extract and a green tea extract,
and the ratio of phenolic compound to hydrogen peroxide can range
from about 3:1 to about 1:3 on a wt/wt basis (molar weight). The
hydrogen peroxide can be added to the phenolic compound using a
concentration of about 0.1% to about 10% hydrogen peroxide
solution. One of skill can easily selected the dose for a
particular use, which will vary according to use, due to
environmental conditions at the site of use. In some embodiments,
this formulation has worked well for uses in animals that are
non-humans.
[0107] As such, the binding system will selectively target damaged
tissues and pathogens infecting those tissues, whereas the same
microbes passively occupying healthy surrounding tissues and
healthy surround tissues themselves will not activate the binding
system. The same type of localized and selective response can be
expected, for example, for inflammations and infections as with
toxins.
[0108] The binding system can be carried in a liquid, powder,
capsule, tablet, or gas for administration to a subject. The
selection of the phenolic compound should take into consideration
the manner in which the reactive oxygen species will bind to the
phenolic compound to form a stable, or substantially stable,
binding pair. The binding pair can be considered substantially
stable where the reactive oxygen species retains all, most, or at
least a predictable amount of oxidation strength for the uses and
functions recited herein.
[0109] One of skill will appreciate that a phenolic compound can be
derivatized to introduce or enhance a desired function. The
phenolic compound can be derivatized, for example, to increase it's
functionality for binding to the reactive oxygen species,
maintaining stability or miscibility in a carrier, or binding to a
target site, using any method known to one of skill. In some
embodiments, the phenolic compound can be bound to a polyol,
pegylated, attached to a saccharide, or attached to glucose, for
example,
[0110] One of skill will appreciate that a phenolic compound can be
linked to another chemical entity by a linker in order to introduce
or enhance a desired function. In some embodiments, a linker can
include, for example, from 1 to 4 amino acids, natural or
synthetic. In some embodiments, a synthetic linker can include an
aminoalkanoic acid having from about 1 to about 20 carbons, from
about 2 to about 14 carbons from about 3 to about 12 carbons, from
about 4 to about 11 carbons, from about 5 to about 10 carbons, or
any range therein. Examples can include, but are not limited to
4-aminobutanoic acid, 5-aminopentanoic acid, 6-aminohexanoic acid,
7-aminoheptanoic acid, 8-amino-octanoic acid, 9-aminononanoic acid
10-aminodecanoic acid, 11-aminoundecanoic, and the like. One of
skill will appreciate that these linkers can be substituted, as
long as the linker functions in accordance with the teachings
provided herein. In some embodiments, the binding system can be
cross linked onto a microbead, magnetic particle, nano-particle or
other substrate to form a reaction enhanced, tissue specific or
steerable ligand or therapeutic system.
[0111] The binding systems can include, for example, a weight ratio
of phenolic compound to reactive oxygen species that ranges from
about 1:1000 to about 1000:1. In some embodiments, the ratio of
phenolic compound to reactive oxygen species can range from about
1:1000 to about 500:1, from about 1:500 to about 500:1, from about
1:250 to about 500:1, from about 1:500 to about 250:1, from about
1:250 to about 250:1, from about 1:100 to about 250:1, from about
1:250 to about 100:1, from about 1:100 to about 100:1, from about
1:100 to about 50:1, from about 1:50 to about 100:1, from about
1:50 to about 50:1, from about 1:25 to about 50:1, from about 1:50
to about 25:1, from about 1:25 to about 25:1, from about 1:10 to
about 10:1, from about 1:1000 to about 250:1, from about 1:1000 to
about 100:1, from about 1:1000 to about 50:1, from about 1:1000 to
about 25:1, from about 1:1000 to about 10:1, from about 1:1000 to
about 5:1, from about 1:10 to about 1:20, from about 1:10 to about
1:30, from about 1:10 to about 1:40, from about 1:10 to about 1:50,
from about 1:10 to about 1:60, from about 1:10 to about 1:70, from
about 1:10 to about 1:80, from about 1:10 to about 1:90, from about
1:20 to about 1:30, from about 1:20 to about 1:40, from about 1:20
to about 1:50, from about 1:20 to about 1:60, from about 1:20 to
about 1:70, from about 1:20 to about 1:80, from about 1:20 to about
1:90, from about 1:30 to about 1:90, or any range therein.
[0112] In some embodiments, the binding system comprises a ratio of
a tannin and hydrogen peroxide, a phenylpropanoid and a hydrogen
peroxide, a catechin and hydrogen peroxide, an epigallic acid and a
hydrogen peroxide, or a combination thereof an of these phenolic
compounds with hydrogen peroxide.
[0113] In some embodiments, the binding systems include a stable
hydrogen bonded complex between the phenolic compound and the
reactive oxygen species. For example, a highly hydroxylated
polyphenol compound can be combined with a high concentration of
hydrogen peroxide, the combination leading to binding the hydrogen
peroxide to the phenolic compound to produce the binding system.
The binding system can be intended for dilution in water or a solid
excipient. One of skill will appreciate that such a complex can be
referred to as a polyphenol peroxysolvate, in some embodiments,
when in a liquid form for storage or administration to a subject,
and a phenolic perhydrate when in an anhydrous, or substantially
anhydrous, form for storage or administration to a subject.
[0114] One of skill will appreciate that the binding systems should
be produced free of compounds that can lead to degradation of the
otherwise stable, or substantially stable, binding pairs. As such,
in some embodiments, the compositions comprise solutes that are
substantially free of transition metals, metal ions, heavy metals,
oxidoreductase enzymes, other strong oxidizers, reactive halogen
compounds, hydrogen halides, and other compounds that can cause a
decomposition of the reactive oxygen species, or its disassociation
from the phenolic compound with which it forms a binding pair.
Methods of Using the Binding Systems
[0115] The compositions taught herein can be used for medicinal
purposes, as a health supplement, or a nutritional composition. The
compositions can provide a therapeutic and/or prophylactic effect
in the treatment of a condition in a subject. The targeted action
of the binding systems allows for the administration of
surprisingly low effective doses of the phenolic compounds. As a
result, the binding systems also improve safety by substantially
increasing the separation between an effective dose and any
toxic/side effects.
[0116] The terms "treat," "treating," and "treatment" can be used
interchangeably and refer to the administering or application of
the binding systems taught herein, including such administration as
a health or nutritional supplement, and all administrations
directed to the prevention, inhibition, amelioration of the
symptoms, or cure of a condition taught herein. The terms
"disease," "condition," "disorder," and "ailment" can be used
interchangeably in some embodiments. The term "subject" and
"patient" can be used interchangeably and refer to an animal such
as a mammal including, but not limited to, non-primates such as,
for example, a cow, pig, horse, cat, dog, rat and mouse; and
primates such as, for example, a monkey or a human. As such, the
terms "subject" and "patient" can also be applied to non-human
biologic applications including, but not limited to, veterinary,
companion animals, commercial livestock, aquaculture, and the like.
Many of the applications can include control environmental
pathogens that are on or in plants, as well as places not
necessarily in living hosts, such as those that are in water and
water systems, for example, as well as soil, air, and food for
microbial control, alteration of surface characteristics, or
anywhere that can benefit from a supply of a stable oxidizer
source.
[0117] In some embodiments, the binding system includes (i) a
phenolic compound selected from the group consisting of condensed
tannins, hydrolysable tannins, complex tannins, phlorotannins,
psuedotannins, and derivatives thereof; and, (ii) hydrogen peroxide
in a stable, or substantially stable, non-covalent association.
When the binding system is combined with an oxidoreductase enzyme
at a target site, the combination promotes increased binding,
complexing, metabolizing or crosslinking of the phenolic compound
with the tissues, pathogens and toxins in or on a subject. The
binding systems can be administered to increase the bioactivity of
the phenolic compound in a binding reaction. It should be noted
that the bioactivity at a GI, dermal, or mucosal target site can be
detrimentally affected by a reduced bioavailability, such as by
absorption rates of the phenolic compound into the systemic
circulation. And, the adverse effects of such absorption on a
subject, the phenolic compounds that create them, and the amounts
at which they occur, remain unknown. It is known, however, that
gallic acid and isoflavones, for example, can be considered as the
most well-absorbed phenols, followed by catechins (flavan-3-ols),
flavanones, and quercetin glucosides, each having different
kinetics. In contrast, the least well-absorbed phenols are the
proanthocyanidins, galloylated tea catechins, and anthocyanins.
[0118] Generally speaking, the binding systems provided herein
selectively bind to, and reduce, the infectivity or propogation of
virus, bacteria, yeast or fungi; and, upon enzymatic bioactivation
by pathogens or damaged tissues, exhibit increased binding
inactivation of endotoxins, such as lipopolysaccharides, and
exotoxins, such as cholera toxin, botulism, and other virulence
factors of bacteria that are pathogenic to a subject, human or
non-human. Likewise, the binding systems exhibit a localized
astringent effect upon a damaged tissue of a subject. Without
intending to be bound by any theory or mechanism of action, this is
believed to be due to the tissue presenting higher levels of
oxidoreductase enzymes than comparable undamaged tissues, making
the action of the binding system serve as a localized and targeted
action that is selective to the damaged tissue. Moreover, the
binding systems function to treat a tissue suffering from
inflammation by reducing the inflammation in, also in a targeted
manner upon bioactivation of the binding system at the target site.
The systems can be used, for example, to treat a GI condition, a
dermal condition, or a mucosal condition. And, it should be
appreciated that, in some embodiments, the binding systems can be
used as a health or nutritional supplement as a prophylactic method
of treatment to prevent the onset of a condition; a treatment.
management, or cure of a condition that has already onset; or a way
to ameliorate the symptoms of such a condition that has already
onset.
[0119] In some embodiments, the binding systems taught herein can
be used to protect, maintain, improve, or restore a digestive
health of a subject when administered orally in an effective
amount, the effectiveness measured by comparing to a control group
that did not receive the binding system. The binding systems can be
used to prevent, inhibit, or ameliorate the symptoms associated
with a loss of digestive tract homeostasis. In some embodiments,
the binding systems can be used to prevent, treat, ameliorate the
symptoms of, or even cure, a chronic gastrointestinal condition.
Such conditions can include, but are not limited to, hyperacidity,
colitis, irritable bowel syndrome, crohn's disease, necrotic
enteritis, functional colonic diseases, malabsorption, a peptic
ulcer, gastro-esophageal reflux disease, ulcerative colitis, and
diverticulitis. In some embodiments, the binding systems can be
used to reduce mucosal tissue inflammation, dysfunction, or damage.
Such conditions can be induced, for example, by drug side effects,
chemotherapy, dysbiosis, radiation, changes in normal flora,
hyperimmunity, autoimmune reactions, immune deficiencies,
nervousness, allergies, chemical irritation, and stress. In some
embodiments, the binding systems can be administered for
selectively inhibiting the growth of gastrointestinal pathogens. It
should be appreciated that there may be lesser inhibition of
non-pathogenic strains, particularly common probiotic bacteria such
as bifidobacteria and lactobacilli. And, in some embodiments,
administration of the binding systems can produce short term immune
modulation effects as well as potentially change the chronic
expression of the activating enzymes associated with some
conditions with longer term use of the binding systems.
[0120] In some embodiments, the symptoms of a gastrointestinal
condition can include, for example, diarrhea, dehydration,
malnutrition, constipation, nausea, and/or cramping. And, in some
embodiments, the symptoms of a gastrointestinal condition can be
temporary and include acid irritation, indigestion, bloating,
cramps, spasmodic peristalsis, diarrhea, and constipation.
Administering the binding systems for the treatment and/or
management of gastrointestinal conditions can be considered a
nutritional or health supplement, in some embodiments. In some such
embodiments, for example, the binding pair can be administered to
prevent, inhibit, or ameliorate the effect, infectivity, and
virulence of pathogens including bacteria, virus, fungi, yeast,
prions, protazoa and parasites in a subject orally taking an
effective amount of the supplement.
[0121] As such, in some embodiments, the teachings are directed to
a system to facilitate an improved bioactivity and increased enzyme
activation rates upon contact of the binding system with damaged
cells, white blood cells or bacterial infection, while remaining
passive to tissues that do not present such enzymes and
non-pathogenic microbiota. In these embodiments, the bioactivation
can be mediated by oxidoreductase enzymes, for example, which
modify phenolic compounds in-situ. The reaction rate can be
limited, for example, by availability of hydrogen peroxide or one
of its degradation products. In some embodiments, the
oxidoreductase enzymes may be native to damaged animal cells or
pathogenic bacteria. The system can, therefore, provide a localized
increase in ability of the phenolic compounds taught herein to form
covalent complexes with a target. The target can include, for
example, amino acids, alcohols, peptides, oligopeptides, proteins,
saccharides, polyols, and the like, as well as other macromolecules
involved with bacterial infection, inflammatory response, tissue
damage, tissue healing.
[0122] The binding systems are also useful in treating wounds.
Generally speaking, the binding systems can protect, seal,
disinfect, promote healing, or improve function of skin or mucosa.
In some embodiments, for example, a wound and a chronic
inflammatory condition can be treated including, but not limit to,
a wound by (i) physical damage, (ii) adiabetic skin lesion, (iii)
abed sore, (iv) a burn, (v) a cold sore, (vi) psoriasis, (vii)
eczema, and (viii) dermatological inflammation caused by pathogens,
to name a few.
[0123] The binding systems are also useful in treating
inflammations. In some embodiments, the binding systems are useful
in treating inflammations of gastrointestinal system, urinary
tract, reproductive system, or respiratory system inflammations in
a subject, in which the binding systems can be administered, for
example, in the form of an enema, nasal spray, or respiratory mist
to prevent, treat, inhibit, or ameliorate the symptoms of an
inflammation of a mucosal tissue.
[0124] The binding systems are also useful in treating infections.
In some embodiments, the binding systems can be used to treat
infections of gastrointestinal system, urinary tract, reproductive
system, or respiratory system infections in a subject, in which the
binding systems can be administered, for example, in the form of an
enema, nasal spray, or respiratory mist to prevent, treat, inhibit,
or ameliorate the symptoms of an infection of a mucosal tissue. In
some embodiments, the binding systems find a particularly useful
application in women, children, and pets.
[0125] In some embodiments, the binding system is in a liquid form
as a general health tonic. Liquid systems can include, but are not
limited to, any liquid formulation known to one of skill. In some
embodiments, the liquid formulation can include a solution, a
colloid, a suspension, an emulsion, a liposomal formulation, and
the like. In some embodiments, the binding system is in a liquid
form for treatment of a short term acute digestive condition.
Examples of such conditions include, but are not limited to,
diarrhea, food poisoning, and traveler's diarrhea. And in some
embodiments, the binding system is in a liquid form for treatment
of a chronic digestive condition. Examples of such conditions
include, but are not limited to, gastroesophageal reflux disease,
inflammatory bowel disease, irritable bowel syndrome, and food
allergies.
[0126] In some embodiments, the binding system is a dry system. For
example, the system can be in the form of a powder, pill, tablet,
capsule, or a separate dry components for mixing into a liquid
form. In these embodiments, both the phenolic compound and the
reactive oxygen species are in a dry form either before or after
creation of the binding pair, and the binding system can be used in
the dry form, or converted to a liquid form, for any of the uses
taught herein. The advantages of the dry compositions can include,
for example, the ease of storage and transport. In some
embodiments, the binding systems, whether in liquid or dry form,
can be combined with vitamins, electrolytes, and/or other nutrients
in either liquid or dry form. The dry form of the binding system
can be manufactured using any drying process known to one of skill,
such as solvent exchange, vacuum drying, critical point drying,
heating, dessication, or a combination thereof. In some
embodiments, the phenolic compound is dried as a single component.
In some embodiments, the binding pair is formed, and the binding
pair is dried together. And, in some embodiments, the reactive
oxygen species can be, independently, in any dry form known to one
of skill, such as the dry forms taught herein. In embodiments
having the reactive oxygen species in an independent dry form, the
dry phenolic compound and the dry reactive oxygen species can be
combined in a polar solvent, for example, to create the binding
pair prior to use.
[0127] The binding systems can be in the form of a kit. In some
embodiments, the kit can comprise a binding system taught herein,
wherein the kit comprises a dry form of the phenolic compound
component and a dry form of the reactive oxygen species component,
as well as instructions for mixing the components to create the
binding system for administration and suggested dilution factors
for various target sites. In some embodiments, the kit can comprise
a dry form of the binding system, as well as instructions for
diluting the binding system for administration with suggested
dilution factors for various target sites. The suggested dilution
factors can be selected from the ranges taught herein.
[0128] As described herein, the binding systems can be used in a
method of treating a damaged dermal, mucosal, or gastrointestinal
tissue. In some embodiments, the method can comprise administering
an effective amount of a binding systema taught herein to the
damaged tissue of the subject. The binding system can function as
an antimicrobial when bioactivated at the target site of the
damaged tissue and assist in the healing of the damaged tissue by
inactivating compounds that promote infection at the target
site.
[0129] As described herein, the binding systems can be used in a
method of treating a gastrointestinal condition. In some
embodiments, the method can comprise administering an effective
amount of a binding system taught herein to the gastrointestinal
tract of the subject. The binding system can function as an
astringent, an anti-toxin, an anti-inflammatory, or an
antimicrobial, for example, when bioactivated at the target site of
the damaged tissue and assists in the healing of the damaged tissue
by inactivating compounds that promote the condition at the target
site.
[0130] As described herein, the binding systems can be used in a
method of treating acute diarrhea in a subject. In some
embodiments, the methods comprise orally administering an effective
amount of a binding system taught herein to the subject. The
binding system can prevent, inhibit, or ameliorate a symptom of
acute diarrhea in the subject when compared to a second subject in
a control group in which the binding system was not administered.
In some embodiments, the symptom is selected from the group
consisting of a stool score, heartburn, indigestion, urgency of
defecation, nausea, vomiting, stomach pain, and bloating.
[0131] As described herein, the binding systems can be used in a
method of promoting weight gain in a subject. In some embodiments,
the method comprises orally administering an effective amount of a
binding system taught herein to the subject as a supplement to the
diet of the subject. The binding systems can increase the feed
conversion ratio of the subject when compared to a second subject
in a control group in which the binding system was not
administered.
[0132] As described herein, the binding systems can be used in a
method of treating irritable bowel syndrome in a subject. In some
embodiments, the method comprises orally administering an effective
amount of a binding system taught herein to the subject. The
binding system can prevent, inhibit, or ameliorate the symptoms of
irritable bowel syndrome in the subject when compared to a second
subject in a control group in which the binding system was not
administered. In some embodiments, the symptom is selected from the
group consisting of a stool score, heartburn, indigestion, urgency
of defecation, nausea, vomiting, stomach pain, and bloating.
[0133] As described herein, the binding systems can be used in a
method of treating an inflammatory bowel disease in a subject. In
some embodiments, the method comprises orally administering an
effective amount of a binding system taught herein to the subject.
The binding system can prevent, inhibit, or ameliorate the symptoms
of inflammatory bowel disease in the subject when compared to a
second subject in a control group in which the binding system was
not administered. In some embodiments, the symptom is selected from
the group consisting of a stool score, heartburn, indigestion,
urgency of defecation, nausea, vomiting, stomach pain, and
bloating.
[0134] As described herein, the binding systems can be used in a
method of treating food poisoning in a subject. In some
embodiments, the method comprises orally administering an effective
amount of a binding system taught herein to the subject. The
binding system can prevent, inhibit, or ameliorate the symptoms of
food poisoning in the subject when compared to a second subject in
a control group in which the binding system was not administered.
In some embodiments, the symptom is selected from the group
consisting of a stool score, heartburn, indigestion, urgency of
defecation, nausea, vomiting, stomach pain, and bloating.
[0135] As described herein, the binding systems can be used in a
method of treating a wound on a tissue of a subject. In some
embodiments, the method comprises administering an effective amount
of a binding system taught herein to a wound of the subject. The
binding system can enhance the rate of healing in the subject when
compared to a second subject in a control group in which the
binding system was not administered. In some embodiments, the wound
is to a dermal tissue, mucosal tissue, or gastrointestinal
tissue.
[0136] As described herein, the binding systems can be used in a
method of improving the gastrointestinal health of in a subject. In
some embodiments, the method comprises orally administering a
binding system taught herein, wherein, the binding system improves
the gastrointestinal health in the subject when compared to a
second subject in a control group in which the binding system was
not administered.
Methods of Administration
[0137] In some embodiments, the binding systems can be administered
to a subject in any non-parenteral manner known to one of skill,
where a parenteral administration involves piercing the skin or a
mucous membrane. In these embodiments, the administration can be
oral, ocular, otologic, nasal, urogenital, rectal, dermal, or to a
mucous membrane. In some embodiments, the administration can be
oral or topical, using any manner of administration known to one of
skill. Oral administration can include digestive tract, buccal,
sublingual, sublabial, and respiratory tract administration, and a
carrier such as a solid or liquid can be used. One of skill will
appreciate that the therapeutic program selected, the agents
administered, the condition of the subject, and the effects
desired, can affect the administration schedule and program
used.
[0138] In many embodiments, the binding systems can be administered
orally in diluted in aqueous solutions, or incorporated with
excipients. The binding systems can be contained in forms that
include tablets, troches, capsules, elixirs, beverages,
suspensions, syrups, wafers, chewing gums, gels, hydrogels, and the
like. Tablets, pills, capsules, troches liquids and the like may
also contain binders, excipients, disintegrating agent, lubricants,
glidants, chelating agents, buffers, tonicity modifiers,
surfactants, sweetening agents, and flavoring agents. Some examples
of binders include microcrystalline cellulose, gum tragacanth or
gelatin. Some examples of excipients include starch or
maltodextrin. Some examples of disintegrating agents include
alginic acid, corn starch and the like. Some examples of lubricants
include magnesium stearate or potassium stearate. An example of a
chelating agent is EDTA. Some examples of buffers are acetates,
citrates or phosphates. Some examples of tonicity modifiers include
sodium chloride and dextrose. Some examples of surfactants for
micellation or increasing cell permeation include coconut soap,
anionic, cationic or ethoxylate detergents. An example of a glidant
is colloidal silicon dioxide. Some examples of sweetening agents
include sucrose, saccharin and the like. Some examples of flavoring
agents include peppermint, chamomile, orange flavoring and the
like. It should be appreciated that the materials used in preparing
these various compositions should be pharmaceutically pure and
non-toxic in the amounts used.
[0139] In the digestive tract, a solid can include a pill, capsule,
tablet, or time-release technology in some embodiments; and, a
liquid can include a solution, soft gel, suspension, emulsion,
syrup, elixir, tincture, or a hydrogel. Digestive tract
administration can include oral or rectal administration using any
method known to one of skill. For buccal, sublingual, and sublabial
administration, a solid can include an orally disintegrating
tablet, a film, a lollipop, a lozenge, or chewing gum; and, a
liquid can include a mouthwash, a toothpaste, an ointment, or an
oral spray.
[0140] For respiratory tract administration, which also includes
any tissue or cavity in communication with the respiratory track,
such as the sinuses, a solid can be administered using a smoking
device; and, a liquid can be administered using a pressurized
metered dose inhaler, a nebulizer, or a vaporizer. In some
embodiments, nasal administration can be used and includes
administering the binding system to the mucus membranes of the
nasal passage or nasal cavity of a subject. Any method of nasal
administration known to one of skill to be suitable for the
compositions provided herein can be used. In some embodiments, the
nasal administration can include nasal spray, nasal drop,
suspension, gel, ointment, cream or powder. In some embodiments, a
nasal tampon or nasal sponge can be used.
[0141] For ocular, otologic, and nasal administrations, the
compounds can be administered using a nasal spray, ear drops, eye
drops, an ointment, a hydrogel, nanosphere suspension, or a
mucoadhesive microdisc. For urogenital administrations, the
compounds can be administered using an ointment, a pessary such as
a vaginal suppository, or a vaginal douche. For rectal
administrations, which also includes administration into the large
intestine in some embodiments, the compounds can be administered
using an ointment, a suppository, an enema, a Murphydrip, a
nutrient enema, or using an endoscopic device. For Dermal
administrations, the compounds can be administered using an
ointment, a liniment, a paste, a film, a hydrogel, liposomes,
transfersome vesicals, cream, lotion, lip balm, medicated shampoo,
a dermal patch, or a dermal spray.
[0142] One of skill understands that the amount of the agents
administered can vary according to factors such as, for example,
the type of disease, age, sex, and weight of the subject, as well
as the method of administration. For example, an administration can
call for substantially different amounts to be effective. Dosage
regimens may also be adjusted to optimize a therapeutic response.
In some embodiments, a single bolus may be administered; several
divided doses may be administered over time; the dose may be
proportionally reduced or increased; or, any combination thereof,
as indicated by the exigencies of the therapeutic situation and
factors known one of skill in the art. It is to be noted that
dosage values may vary with the severity of the condition to be
alleviated, as well as whether the administration is prophylactic,
such that the condition has not actually onset or produced
symptoms. Dosage regimens may be adjusted over time according to
the individual need and the professional judgment of the person
administering or supervising the administration of the
compositions, and the dosage ranges set forth herein are exemplary
only and do not limit the dosage ranges that may be selected by
medical practitioners.
[0143] The terms "administration" or "administering" can be used to
refer to a method of incorporating a composition into the cells or
tissues of a subject, either in vivo or ex vivo to test the
activity of a system, as well as to diagnose, prevent, treat, or
ameliorate a symptom of a disease. In one example, a compound can
be administered to a subject in vivo using any means of
administration taught herein. In another example, a compound can be
administered ex vivo by combining the compound with cell tissue
from the subject for purposes that include, but are not limited to,
assays for determining utility and efficacy of a composition. And,
of course, the binding systems can be used in vitro to test their
stability, activity, toxicity, efficacy, and the like. When the
compound is incorporated in the subject in combination with one or
active agents, the terms "administration" or "administering" can
include sequential or concurrent incorporation of the compound with
the other agents such as, for example, any agent described above. A
pharmaceutical composition of the invention can be formulated, in
some embodiments, to be compatible with its intended route of
administration.
[0144] An "effective amount" of a compound can be used to describe
a therapeutically effective amount or a prophylactically effective
amount. An effective amount can also be an amount that ameliorates
the symptoms of a disease. A "therapeutically effective amount" can
refer to an amount that is effective at the dosages and periods of
time necessary to achieve a desired therapeutic result and may also
refer to an amount of active compound, prodrug or pharmaceutical
agent that elicits any biological or medicinal response in a
tissue, system, or subject that is sought by a researcher,
veterinarian, medical doctor or other clinician that may be part of
a treatment plan leading to a desired effect. In some embodiments,
the therapeutically effective amount should be administered in an
amount sufficient to result in amelioration of one or more symptoms
of a disorder, prevention of the advancement of a disorder, or
regression of a disorder. In some embodiments, for example, a
therapeutically effective amount can refer to the amount of an
agent that provides a measurable response of at least 5%, at least
10%, at least 15%, at least 20%, at least 25%, at least 30%, at
least 35%, at least 40%, at least 45%, at least 50%, at least 55%,
at least 60%, at least 65%, at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, or at least 100% of
a desired action of the composition.
[0145] A "prophylactically effective amount" can refer to an amount
that is effective at the dosages and periods of time necessary to
achieve a desired prophylactic result, such as prevent the onset of
an inflammation, allergy, nausea, diarrhea, infection, and the
like. Typically, a prophylactic dose is used in a subject prior to
the onset of a disease, or at an early stage of the onset of a
disease, to prevent or inhibit onset of the disease or symptoms of
the disease. A prophylactically effective amount may be less than,
greater than, or equal to a therapeutically effective amount.
[0146] Any administration vehicle known to one of skill to be
suitable for administration of the compounds, compositions, and
formulations taught herein can be used. A "vehicle" can refer to,
for example, a diluent, excipient or carrier with which a compound
is administered to a subject.
[0147] The compounds can be administered in dosage units. The term
"dosage unit" can refer to discrete, predetermined quantities of a
compound that can be administered as unitary dosages to a subject.
A predetermined quantity of active compound can be selected to
produce a desired therapeutic effect and can be administered with a
pharmaceutically acceptable carrier. The predetermined quantity in
each unit dosage can depend on factors that include, but are not
limited to, (a) the unique characteristics of the active compound
and the particular therapeutic effect to be achieved, and (b) the
limitations inherent in the art of creating and administering such
dosage units.
[0148] A "pharmaceutically acceptable carrier" is a diluent,
adjuvant, excipient, or vehicle with which the composition is
administered. A carrier is pharmaceutically acceptable after
approval by a state or federal regulatory agency or listing in the
U.S. Pharmacopeial Convention or other generally recognized sources
for use in subjects.
[0149] The pharmaceutical carriers include any and all
physiologically compatible solvents, dispersion media, coatings,
antibacterial and antifungal agents, isotonic and absorption
delaying agents, and the like. Examples of pharmaceutical carriers
include, but are not limited to, sterile liquids, such as water,
oils and lipids such as, for example, phospholipids and
glycolipids. These sterile liquids include, but are not limited to,
those derived from petroleum, animal, vegetable or synthetic origin
such as, for example, peanut oil, soybean oil, mineral oil, sesame
oil, and the like.
[0150] Suitable pharmaceutical excipients include, but are not
limited to, starch, sugars, inert polymers, glucose, lactose,
sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium
stearate, glycerol monostearate, talc, sodium chloride, dried skim
milk, glycerol, propylene, glycol, water, ethanol, and the like.
The composition can also contain minor amounts of wetting agents,
emulsifying agents, pH buffering agents, or a combination thereof.
The compositions can take the form of solutions, suspensions,
emulsion, tablets, pills, capsules, powders, sustained-release
formulations and the like. Oral formulations can include standard
carriers such as, for example, pharmaceutical grades mannitol,
lactose, starch, magnesium stearate, sodium saccharine, cellulose,
magnesium carbonate, and the like. See Martin, E.W. Remington's
Pharmaceutical Sciences. Supplementary active compounds can also be
incorporated into the compositions.
[0151] In some embodiments, an administration, such as an oral or
topical administration, may include liposomes. In some embodiments,
the liposome may assist in a targeted delivery system. The
liposomes can be designed, for example, to bind to a target protein
and be taken up selectively by the cell expressing the target
protein.
[0152] Therapeutic compositions typically must be sterile and
stable under the conditions of manufacture and storage. The
composition can be formulated as a solution, microemulsion,
liposome, or other ordered structure suitable for a desired
concentration of the compound. In some embodiments, the carrier can
be a solvent or dispersion medium including, but not limited to,
water; ethanol; a polyol such as for example, glycerol, propylene
glycol, liquid polyethylene glycol, and the like; and, combinations
thereof. The proper fluidity can be maintained in a variety of ways
such as, for example, using a coating such as lecithin, maintaining
a required particle size in dispersions, and using surfactants.
[0153] In some embodiments, isotonic agents can be used such as,
for example, sugars; polyalcohols that include, but are not limited
to, mannitol, sorbitol, glycerol, and combinations thereof; and
sodium chloride. Sustained absorption characteristics can be
introduced into the compositions by including agents that delay
absorption such as, for example, monostearate salts, gelatin, and
slow release polymers. Carriers can be used to protect against
rapid release, and such carriers include, but are not limited to,
controlled release formulations in implants and microencapsulated
delivery systems. Biodegradable and biocompatible polymers can be
used such as, for example, ethylene vinyl acetate, polyanhydrides,
polyglycolic acid, collagen, polyorthoesters, polylactic acid,
polycaprolactone, polyglycolic copolymer, and the like. Such
formulations can generally be prepared using methods known to one
of skill in the art.
[0154] The compounds may be administered as suspensions or
emulsions. Lipophilic solvents or vehicles include, but are not
limited to, fatty oils such as, for example, sesame oil; synthetic
fatty acid esters, such as ethyl oleate or triglycerides; and
liposomes. Suspensions that can be used for injection may also
contain substances that increase the viscosity of the suspension
such as, for example, sodium carboxymethyl cellulose, sorbitol, or
dextran. Optionally, a suspension may contain stabilizers or agents
that increase the solubility of the compounds and allow for
preparation of highly concentrated solutions.
[0155] In some embodiments, a therapeutically or prophylactically
effective amount of a composition may range in concentration from
about 0.01 nM to about 0.10 M; from about 0.01 nM to about 0.5 M;
from about 0.1 nM to about 150 nM; from about 0.1 nM to about 500
.mu.M; from about 0.1 nM to about 1000 nM, 0.001 .mu.M to about
0.10 M; from about 0.001 .mu.M to about 0.5 M; from about 0.01
.mu.M to about 150 .mu.M; from about 0.01 .mu.M to about 500 .mu.M;
from about 0.01 .mu.M to about 1000 nM, or any range therein. In
some embodiments, the compositions may be administered in an amount
ranging from about 0.005 mg/kg to about 100 mg/kg; from about 0.005
mg/kg to about 400 mg/kg; from about 0.01 mg/kg to about 300 mg/kg;
from about 0.01 mg/kg to about 250 mg/kg; from about 0.1 mg/kg to
about 200 mg/kg; from about 0.2 mg/kg to about 150 mg/kg; from
about 0.4 mg/kg to about 120 mg/kg; from about 0.15 mg/kg to about
100 mg/kg, from about 0.15 mg/kg to about 50 mg/kg, from about 0.5
mg/kg to about 10 mg/kg, or any range therein, wherein a human
subject is often assumed to average about 70 kg.
[0156] In some embodiments, the compounds can be administered by
inhalation through an aerosol spray or a nebulizer that may include
a suitable propellant such as, for example,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide, or a combination
thereof. In one example, a dosage unit for a pressurized aerosol
may be delivered through a metering valve. In another embodiment,
capsules and cartridges of gelatin, for example, may be used in an
inhaler and can be formulated to contain a powderized mix of the
compound with a suitable powder base such as, for example, starch
or lactose.
[0157] Rectal administrations can be made using any method known to
one of skill. For example, a suppository formulation can be
prepared by heating glycerin to about 120.degree. C., combining the
binding system with the heated glycerin, mixing the combination,
adding purified water to a desired consistency, and pouring the
desired consistency into a mold to form the suppository.
[0158] For topical administration, suitable formulations may
include a biocompatible oil, wax, gel, powder, emulsion, polymer,
or other liquid or solid carriers. Such formulations may be
administered by applying directly to affected tissues. For example,
a liquid formulation to treat infection of aural canal can be
administered dropwise into the subject's ear. In another example, a
hydrogel infused with the binding system can be applied to a burn.
In another example, a cream formulation can be administered to an
area of psoriasis. Transdermal administration includes percutaneous
absorption of the composition through the skin. Transdermal
formulations include patches, ointments, creams, gels, salves, and
the like.
[0159] In some embodiments, the binding system is administered in a
sustained release formulation, and the formulation can include one
or more agents in addition to the binding system. In some
embodiments, the sustained release formulations can reduce the
dosage and/or frequency of the administrations of such agents to a
subject. In some embodiments, an exogenous catalyst or enzyme is
introduced to a target and one or more of the reactive oxygen
species, phenolic compound, or the exogeneous catalyst or enzyme
are segregated by encapsulation or micellation to delay the
bioactivation until target site is reached by all components.
[0160] The amount of the compound administered may vary widely
depending on the type of formulation, size of a unit dosage, kind
of excipients, and other factors well known to those of ordinary
skill in the art. The formulation may comprise, for example, from
about 0.0001% to about 6% (w/w), from about 0.01% to about 1%, from
about 0.1% to about 0.8%, or any range therein, with the remainder
comprising the excipient or excipients.
[0161] In some embodiments, the composition can be administered in
conjunction with at least one other therapeutic agent for the
condition being treated. The amounts of the agents can be reduced,
even substantially, such that the amount of the agent or agents
desired is reduced to the extent that a significant response is
observed from the subject. A significant response can include, but
is not limited to, a reduction or elimination of nausea, a visible
increase in tolerance, a faster response to the treatment, a more
selective response to the treatment, or a combination thereof.
[0162] In some embodiments, the compounds, compositions, and
formulations can be administered in combination with a composition
taught herein using any amount, time, and method of administration
known to be effective by one of skill. The compound can be
administered, for example, in an amount ranging from about 0.1
.mu.g/kg to about 1 mg/kg, from about 0.5 .mu.g/kg to about 500
.mu.g/kg, from about 1 .mu.g/kg to about 250 .mu.g/kg, from about 1
.mu.g/kg to about 100 .mu.g/kg from about 1 .mu.g/kg to about 50
.mu.g/kg, or any range therein. One of skill can readily select the
frequency and duration of each administration.
[0163] In some embodiments, the methods taught herein can further
include the administration of an effective amount of an additional
bioactive agent or therapeutic treatment. In some embodiments, the
terms "agent" and "therapy" can be interchangeable. In many
embodiments, the molecular weight of an agent should be at or below
about 40,000 Daltons to ensure elimination of the agent from a
subject. In some embodiments, the molecular weight of the agent
ranges from about 300 Daltons to about 40,000 Daltons, from about
8,000 Daltons to about 30,000 Daltons, from about 10,000 Daltons to
about 20,000 Daltons, or any range therein.
[0164] Combinations therapies can be administered, for example, for
30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 12 hours, 18 hours,
1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9
days, 10 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 3 months, 6
months 1 year, any combination thereof, or any amount of time
considered necessary by one of skill. The agents can be
administered concomitantly, sequentially, or cyclically to a
subject. Cycling therapy involves the administering a first agent
for a predetermined period of time, administering a second agent or
therapy for a second predetermined period of time, and repeating
this cycling for any desired purpose such as, for example, to
enhance the efficacy of the treatment. The agents can also be
administered concurrently. The term "concurrently" is not limited
to the administration of agents at exactly the same time, but
rather means that the agents can be administered in a sequence and
time interval such that the agents can work together to provide
additional benefit. Each agent can be administered separately or
together in any appropriate form using any appropriate means of
administering the agent or agents.
[0165] As described herein, a stabilized reagent pair can be
administered for aqueous transport to a target site. In some
embodiments, the reagent pair comprises a tannin having a molecular
weight ranging from about 500 Daltons to about 4000 Daltons;
and,
[0166] hydrogen peroxide. The hydrogen peroxide can be hydrogen
bonded to the tannin at a tannin:peroxide weight ratio that ranges
from about 1:1000 to about 10:1; the binding system can be
bioactivated at a target site having an oxidoreductase enzyme; and,
the binding molecule binds to the target site. In some embodiments,
a pharmaceutical formulation comprising a reagant pair taught
herein can be used in an administration, and a pharmaceutically
acceptable excipient. The tannin can comprise a catechin, and the
tannin:peroxide ratio can ranges from about 1:10 to about 1:50. In
some embodiments, the oxidoreductase can comprise a peroxidase;
and, there can be no, or substantially no, unbound hydrogen
peroxide in the formulation.
Articles of Manufacture
[0167] The present invention provides for articles of manufacture
that encompass finished, packaged and labelled products. The
articles of manufacture include the appropriate unit dosage form in
an appropriate vessel or container such as, for example, a glass
vial or other container that is hermetically sealed. In the case of
dosage forms suitable for oral administration, the active
ingredient, e.g. one or more agents including a dosage form taught
herein, may be suitable for administration orally, rectally,
vaginally, or the like. Alternatively, the unit dosage form may be
a solid suitable for oral, transdermal, topical or mucosal
delivery.
[0168] In some embodiments, the unit dosage form is suitable for
oral or topical delivery. Thus, the invention encompasses
solutions, which are preferably stable or substantially stable,
sterile, and suitable for such administrations. The concentration
of agents and amounts delivered are included as described
herein.
[0169] As with any such product, the packaging material and
container are designed to protect the stability of the product
during storage and shipment. In addition, the articles of
manufacture can include instructions for use or other information
material that can advise the user such as, for example, a
physician, technician or patient, regarding how to properly
administer the composition as a prophylactic, therapeutic, or
ameliorative treatment of the disease of concern. In some
embodiments, instructions can indicate or suggest a dosing regimen
that includes, but is not limited to, actual doses and monitoring
procedures.
[0170] In some embodiments, the instructions can include
informational material indicating how to administer the binding
systems for a particular use or range of uses, as well as how to
monitor the subject for positive and/or negative responses to the
binding systems.
[0171] In some embodiments, the articles of manufacture can
comprise one or more packaging materials such as, for example, a
box, bottle, tube, vial, container, sprayer, insufflator,
intravenous (I.V.) bag, envelope, and the like; and at least one
unit dosage form of an agent comprising an extract taught herein
within the packaging material. In some embodiments, the articles of
manufacture may also include instructions for using the composition
as a prophylactic, therapeutic, or ameliorative treatment for the
disease of concern.
[0172] In some embodiments, the articles of manufacture can
comprise one or more packaging materials such as, for example, a
box, bottle, tube, vial, container, sprayer, insufflator, envelope,
and the like; and a first composition comprising at least one unit
dosage form of an agent comprising a binding system as taught
herein within the packaging material, along with a second
composition comprising a second agent such as, for example, any
other bioactive agent that may be administered in combination with
the binding system, or any prodrugs, codrugs, metabolites, analogs,
homologues, congeners, derivatives, salts, solvates, and
combinations thereof. In some embodiments, the articles of
manufacture may also include instructions for using the composition
as a diagnostic, prophylactic, therapeutic, or ameliorative
treatment for the condition of concern.
[0173] In some embodiments, the article of manufacture can include
a substantially anhydrous binding system. For example, a kit can be
assembled which includes the anhydrous binding system comprising an
anhydrous tannin with instructions combining the tannin with and an
anhydrous reactive species generating component that forms a
therapeutically, prophylactically, or nutritionally useful
composition upon hydration.
[0174] Without intending to be limited to any theory or mechanism
of action, the following examples are provided to further
illustrate the teachings presented herein. It should be appreciated
that there are several variations contemplated within the skill in
the art, and that the examples are not intended to be construed as
providing limitations to the claims.
Example 1
Making a Binding System of Hydrolysable Tannin Bound to Hydrogen
Peroxide and Showing a Stable, or Substantially Stable, Binding
Pair
[0175] Chinese Gall is an excellent source of a hydrolysable
tannin. Chinese Gall (GALLAE CHINENSES from the Rhus semialata
galls), contains 60% to 75% tannic acids and 2% to 4% of gallic
acid. Gall extracts characteristically do not contain significant
flavanoids. The polygalloyl glucoses or polygalloyl quinic acid
esters presenting 2-12 gallate residues with a relatively open and
conformable steric arrangement are favorable for forming stable
multiple hydrogen bonds with hydrogen peroxide.
[0176] In this experiment, 1 to 10 grams a serial different
quantity of gallotannic acid from Chinese Gall (Sigma-Aldrich
Chinese Gall) was dissolved in 20 cc of 35% food grade hydrogen
peroxide. Comparisons of oxidizing potential were made
colorimetrically using WATERWORKS peroxide check strips (Industrial
Test Systems, Inc., Rock Hill, S.C.). The solution was desiccated
by heating at 80.degree. C. until the solution was a dark highly
viscous mass. Half of the solution was reconstituted to its
original volume. After 2 hr equilibration time. measurement of
oxidative potential of this solution showed less than 10%
difference from pre-dessicated state, indicating preferential
binding. A minimum molar ratio of H.sub.2O.sub.2 to tannin
compounds required to retain greater than 90% of H.sub.2O.sub.2
potential was used to define an optimal ratio. We find that this
minimum molar ratio varies significantly with the choice and/or
combination of phenolic compounds.
[0177] The other half of the solution was placed in cold
temperature (ice bath) until a precipitate formed. After
centrifuging and removing 50 cc of liquid containing the
precipitate from the solution and returning the samples to room
temperature, the balance of the solution showed significantly lower
peroxide concentration than can be accounted for by the fluid
volume removed. Adding back the 50 cc liquid containing the
precipitate restored free peroxide levels to original levels,
clearly indicating the incorporation of a high concentration of
hydrogen peroxide on the precipitate.
Example 2
Comparing Binding Systems Using a Hydrolysable Tannin, a Condensed
Tannin, a Mixture of Hydrolysable and Condensed Tannins, and
Resveratrol Bound to Hydrogen Peroxide to Compare the Binding
Pairs
[0178] Hydrolysable Tannin--For this example, the Chinese gall of
Example 1 was used as the hydrolysable tannin, in addition to the
following:
[0179] Condensed Tannin--Green tea (Camilla Sinensis) extract
contains catechins and other flavanoid compounds but
characteristically does not contain significant tannic acid
content. Multiple gallate and catechol residues of various catechin
and flavanol dimmers, trimers oligomers and polymers are favorable
structures for stable hydrogen peroxide aggregate formation, though
the flavan structure is more likely to cause steric blocking than
the gallotannic structure.
[0180] Mix of Hydrolysable and Condensed Tannin--Pomegranate POMx
(Punica granatum L., POM Wonderful brand) extract of fruit residue
after pressing containing 86.0% ellagitannins, The approximate
distribution of polyphenols is 19% ellagitannins as punicalagins
and punicalins, 4% free ellagic acid, and 77% heterogenous
oligomers of gallic acid, ellagic acid, and glucose with 2-8 phenol
moieties. The planar structure of the punicaligins and the
generally high number of gallate residues on the ellagitannins
provide abundant opportunities for stable hydrogen
perhydration.
[0181] Resveratrol from polygonum cuspidatum (NutraBio
99.87%--standardized to 50% active trans-resveratrol) a stilbenoid
monomer with only three hydroxyls and low water solubility (0.003
g/l). It has a low binding site ratio of 0.013. Monomers, and lower
molecular weight phenolics with separated hydroxyl groups such as
resorcinol moieties are unfavorable structures for stable
perhydrate formation.
[0182] In order to compare the samples provided above, a test
series was prepared in 30 ml tubes containing from 1 g.about.10 g
of the above extracts of Chinese Gall, Green Tea, Pomegranate and
Resveratrol. Each was dissolved in 20 ml of 35% hydrogen peroxide
then separate to two aliquots. One part was heated at 80.degree. C.
to a dark viscous semi-liquid and allowed to desiccate to a final
volume of 5 ml. Each was rehydrated and serially diluted to
detection range. Hydrogen peroxide colorimetric strips showed
qualitatively different concentrations of hydrogen peroxide were
retained by the different types of polyphenolic compounds.
[0183] Chinese gall and pomegranate extracts showed the highest
peroxide retention capability, green tea extracts also showed good
retention (approximately 1/2) and the resveratrol showed relatively
less ability to form stable perhydrates. The results support the
hypothesized molecular characteristics for formation of useful
binding systems.
[0184] The other aliquot of was placed in an ice bath to
precipitate the binding systems. After centrifuging and removing
the precipitate to a separate tube and re-dissolved in 10 ml of
water, the material went into solution, but the initial level of
oxidation was surprisingly below detection limits. Measurements
taken every 10 minutes showed a gradual increase of oxidation,
reaching equivalence to the other aliquot after approximately 50
minutes. This was determined using WATERWORKS peroxide check strips
(Industrial Test Systems, Inc., Rock Hill, S.C.). This demonstrates
that the binding systems are not covalent complexes and can also be
used as a timed release medium.
Example 3
Data Showing Enzyme Selectivity and Targeting
[0185] A key aspect of the invention is that polyphenol-hydrogen
peroxide aggregates are generally nonreactive with digestive
enzymes such as proteases and peptidases that split proteins into
their monomers, the amino acids, lipases that split fat into three
fatty acids and a glycerol molecule, carbohydrases that split
carbohydrates such as starch and sugars into simple sugars, or
nucleases hat split nucleic acids into nucleotides.
[0186] Binding systems responding to target specific enzymes
exhibit orders of magnitude (500.times. or more) differential
between active and passive states providing focused toxin binding,
pathogen or damage specific effects with a reduction in undesirable
collateral effects. In the animal body, the activated binding
systems can actively form glycosydic bonds, as well as complex
proteins and amino acids. The binding of the phenolic compound to,
for example, glucuronic acid or other glucose moieties can
neutralize the activity of lipopolysaccharides and other important
toxins.
[0187] In this experiment, first, a serial dilution of a binding
system of Chinese Gall-hydrogen peroxide (from 0 to 10 .mu.g/ml)
was incubated with a lipopolysaccharide, then reacted with standard
polymixin B with and without horseradish peroxidase at 37.degree.
C. The result showed that, when combined with horseradish
peroxidase, the Chinese Gall-hydrogen peroxide binding system
exhibited over 500.times. increase in lipopolysaccharide binding
compared to the composition without horseradish peroxidase as
determined by ELISA measurements of polymixin B binding inhibition
test.
[0188] Next, we performed an anti-cholera toxin B antibody binding
inhibition experiment. A serial dilution of a binding system of
Chinese Gall-hydrogen peroxide (from 0 to 10 .mu.g/ml) was combined
with cholera toxin, then reacted with anti-cholera toxin B antibody
with and without horseradish peroxidase at 37.degree. C. The result
showed that the combination of horseradish peroxidase and the
Chinese Gall-hydrogen peroxide binding system exhibited over
500.times. increase over the composition without the peroxidase in
anti-cholera toxin B antibody binding as determined by ELISA
measurements.
[0189] These results clearly demonstrate a surprising and
extraordinarily efficient binding of two distinctly different
toxins upon enzyme activation. The large differential in activity
indicates the viability of delivering a stable polyphenol
perhydrate for localized and aggessive remote activation by
tissues, tissue conditions, or pathogens that express peroxidase
enzymes or other site specific enzymes utilizing hydrogen peroxide
or its decomposition products as a reaction promoting
substrate.
Example 4
Data Showing that an Increase in Bound H.sub.2O.sub.2 on Chinese
Gall Results in a Higher Inhibitory Activity of the Chinese
Gall
[0190] Sample A, which contained 100 mg of Chinese gall (Sigma
Aldrich, Chinese Gall) dissolved in 100 ml of 10% hydrogen
peroxide, then diluted to a total volume of 1000 ml was compared
against Sample B, which contained 1 mg of Chinese gall dissolved in
100 ml of 0.1% hydrogen peroxide (a dilution of the 10% hydrogen
peroxide by 100.times.) and was then also diluted to 1000 ml. Due
to the dilution, the molarity of the diluted hydrogen peroxide was
1/100 of the sample A solution, such that 100.times. less
H.sub.2O.sub.2 was available to bind with the Chinese gall. It was
suspected that a proportionally lower amount of H.sub.2O.sub.2
would be bound on Sample A as compared to Sample B. To compare the
relative effects of the different amounts of available
H.sub.2O.sub.2, the activity of the two binding systems was
compared using a binding assay. Sample A had a higher inhibition
effect, showing that the higher amount of available H.sub.2O.sub.2
resulted in a higher activity of the phenolic compound in the
binding system.
Example 5
Data Showing Treatment of Diarrhea
[0191] Data has shown that the binding systems can protect,
improve, maintain or restore body homeostasis, especially
gastrointestinal health. The binding systems provide antisecretory,
anti-infective, anti-pathogenic, anti-adhesion, anti-allergenic and
anti-toxin functions; as well as promote a localized tissue barrier
formation, tissue healing, gross permeability reduction,
astringency, and a restoration of homeostasis.
[0192] This example illustrates how the binding systems can restore
gastrointestinal health through these overlapping damage specific
functions to synergistically defeat pathogen defenses without
involving typical antibiotic resistance mechanisms. The binding
system was shown to provide a highly effective resolution of
infections and the disruption of homeostasis caused by a microbial
diarrhea.
[0193] In a double blind test, 86 human subjects suffering from
moderate to severe acute diarrhea. The subjects were given either a
binding system or placebo on the first day, and then the opposite
on the next day. The binding system contained less than 5 milligram
dry weight equivalent of a binding system of a mixture of
pomegranate and green tea extracts with hydrogen peroxide. The time
to resolution (last loose stool) was 7 hours with a P<0.06. 43%
of the subjects receiving active product on the first day had no
further loose stools after single dose. Most subjects also reported
rapid cessation of discomfort symptoms.
[0194] This example shows that the binding systems can treat
digestive health conditions associated with pathogen colonization,
toxins, overgrowth of bacteria (dysbiosis) or fungal organisms
(Candida).
Example 5
Data Showing Treatment of a Chronic Candida albicans Infection with
Related Symptoms
[0195] A 42 year old Male with a diagnosed chronic Candida Albicans
infection, or intestine expressing also as skin rashes, experienced
significant reduction of both the rash and abdominal discomfort
after 5 days of ingesting a pomegranate/green tea binding system
with hydrogen peroxide. The symptoms gradually returned to original
severity over 2 weeks after termination of regimen.
[0196] This example shows that the binding systems can not only
treat a GI condition, but they can also reduce symptoms associated
with the GI condition. Such symptoms can include, but are not
limited to, inflammation, sepsis, allergic reaction, pain,
cramping, intestinal spasms, stomach upset, acid irritation,
diarrhea, constipation, bloating, nausea and fatigue.
Example 6
Data Showing Treatment of GI Condition with Near-Immediate
Relief
[0197] 43 adults in a placebo-controlled 24 hr crossover study were
given a 25 ml solution of a green tea/pomegranate binding system
with hydrogen peroxide and observed for 2 hrs after ingestion. The
subjects reported significant reduction in upper gastric acid
discomfort, nausea, bloating and abdominal pain within 2 hrs of
active ingestion vs no notable reduction on placebos. P<0.05 in
all categories.
Example 7
Data Showing Treatment of a Variety of GI Conditions
[0198] In a study of adult subjects given the binding system, the
subjects reported benefits related to treatment of ulcers,
fistulas, irritable bowel syndrome, acid reflux, food poisoning,
inflammatory bowel diseases, food sensitivity, travelers diarrhea,
dietary change, and physical agitation (i.e, agitation to GI track
from running).
[0199] 46 volunteer subjects not experiencing acute diarrhea, but
experience frequent symptoms such as those in FIG. 6b though 6f
(including 6 persons with diagnosed IBS or IBD) ingested the
polyphenol/peroxide binding composition as needed for relief of
symptoms. 78% reported significant benefit.
[0200] The success in such a wide variety of GI conditions suggests
that the binding system may also be helpful in treating the GI
symptoms and conditions related to the administration of
chemotherapy and radiation therapy. Also, the binding systems would
appear to be useful in the treatment of chronic gastrointestinal
conditions including, but not limited to, colitis, irritable bowel
syndrome, Crohn's disease, necrotic enteritis, functional colonic
diseases, malabsorbtion, peptic ulcer, gastro-esophogeal reflux
disease, ulcerative colitis, diverticulitis, and ameliorating their
symptoms.
Example 8
Data Showing Treatment of an Immune-Response GI Condition
[0201] The binding systems can efficiently bind, block, or
neutralize inflammatory agents, as well as immune complements,
antibodies and receptors. This activity facilatates modulating
animal inflammatory response to biotic and abiotic factors,
including reducing autoimmune activity. Bacteria can influence the
phenomenon known as oral tolerance, in which the immune system is
less sensitive to an antigen, including those produced by gut
bacteria, once it has been ingested. This tolerance, mediated in
part by the gastrointestinal immune system and in part by the
liver, can induce a hyper-reactive immune response like those found
in allergies and auto-immune disease.
[0202] Some suspect that inflammation in inflammatory bowel
disease, for example, is due to increased permeability of the inner
lining of the colon. This permeability may allow bacteria to invade
the tissues and cause an immune reaction that leads to prolonged
inflammation. Tissue damage in inflammatory bowel disease results
from the immunological misperception of danger within the naturally
occurring flora or a failure of normal tolerance to pathogenic
bacteria. It is still unclear whether the inflammation that occurs
is due to a specific subset of intestinal microbes or due to a
problem with the tolerance of commensal gut flora. Abnormal leaky
cellular junctions, which are supposed to prevent permeability,
have been found in the cells of patients with inflammatory disease.
Several studies have reported the inhibitory effect of green tea
catechins. For example, epicatechin gallate (ECG) and
eligallocatechin gallate (EGCG) can be incorporated into a binding
system for oral or distal delivery to the intestinal tract to
provide greater anti-inflammatory effect than EGCG or EGC
alone.
[0203] In order to support this theory that the binding systems can
treat such an immune response GI condition, several volunteers were
treated. The volunteers experienced symptoms that suggested such an
immune response problem. They had frequent painful lower abdominal
pain, and they ingested a 1 milligram dry weight equivalent of a
pomegranate/hydrogen peroxide binding system formulation for 5
consecutive days. All reported significant reduction in pain with a
continuing effect lasting for 2-5 days after the last dose.
Example 9
Data Showing Treatment of a GI Condition Relating to an Innate
Immune Response
[0204] Most allergy symptoms tie to the innate immune system.
Sometimes the body over responds to allergens by releasing excess
amounts of histamine, serotonin, prostaglandin, interleukins, etc
causing allergy symptoms. Because of the structural and behavioral
similarities of certain portions of these immune molecules to
phenolic compounds or proteins, an enzyme activated binding system
can have the potential to directly complex to, and inactivate,
immune response compounds or inhibit their receptors.
[0205] In order to support this theory that the binding systems can
treat such an innate immune response in the GI tract, several
people with frequent food allergies were treated. The allergies
related to gluten, dairy and unidentified compounds, and the
subjects were expressing a variety of symptoms such as headaches,
diarrhea, bloating, nausea, rash, or fatigue, anecdotally reported
a consistent reduction or elimination of symptoms after ingestion
of the binding system.
Example 10
Data Showing Topical Treatment of a Dermal Wound
[0206] Without intending to be bound by any theory or mechanism of
action, it is believed that the binding systems can facilitate the
wound healing by at least two mechanisms. The first mechanism is
the activation of binding system at the wounded tissue by the
peroxidase from the damaged tissue. This activation will initiate
the release of reactive oxygen and oxygen molecule to either damage
the potential harmful pathogens at wounded site or initiate the
crosslinking or binding function to neutralize the toxin and
interfere with the pathogen's normal growth function to reduce the
potential infection. The second mechanism is the rapid crosslinking
of damaged tissue surface with a similar function to protein
crosslinking mechanisms during the normal growth and healing
process. The astringent effects and rapid formation of a refractory
barrier by the binding system help to reduce fluid loss and act as
a substrate for facilitate faster healing of the epithelial
tissue.
[0207] In order to support this theory, a controlled wound healing
test was done by providing 0 to 20 .mu.g/ml of a perhydrated green
tea extract directly to bilateral lancet wounds on the backs of
nude mice. Sub-dermal healing was measured electronically using a
BioelectricMed skin potential scanner and visual observation. The
healing time was 3.times. lower than the healing time of comparison
Neosporin treated wounds and equivalent to the healing time
observed using a O2Cure hyperbaric oxygen emulsion.
[0208] To further support this theory, a 10 ug/mg solution of a
green tea extract/peroxide binding system was applied by spraying
twice daily to the injuries of a 62 year old man with full depth
skin abrasions on calf and thigh. Exudate from the injuries
substantially stopped within 12 hours. Epithelialization was 95%
complete within 21 days, and a 3 month follow-up showed only minor
discoloration, as well as normal hair follicles and skin
texture.
[0209] To further support this theory, a 12 year old boy with a
large 2.sup.nd degree burn on his calf and a 9 year old girl with a
2.sup.nd degree burn on her upper arm. Both subjects exhibited
cessation of exudation from the burns within one day of application
and unusually rapid epithelialization. The wounds healed without
visible infection or scarring.
[0210] This experiment shows that the binding systems can
facilitate the wound healing process for cuts, abrasions, and burns
of dermal tissue.
Example 11
Data Showing Topical Treatment of an Inflammatory Condition
[0211] The synergistic combination of antimicrobial,
anti-inflammatory and tissue repair effects presented by direct or
indirect application of the binding systems to compromised tissues
have valuable application in correcting abnormal conditions on any
dermal, epidermal tissue or mucosal tissue. These include
inflammatory or autoimmune conditions of the alimentary canal,
urinary tract, reproductive tract, respiratory tract, sinuses,
aural canal, tear ducts, peritoneum and skin.
[0212] To illustrate the applicability of the binding systems to
inflammatory conditions, anecdotal observation of complete and
permanent resolution of long standing scaly psoriasis isolated to
the legs and hands, face or scalp of 5 individuals after direct
topical application of pomegranate/green tea extract binding system
with hydrogen peroxide for 7 days. Twice daily, a spray of 20 ug/ml
solution was administered and caused the scale to begin sloughing
off within 2 days with significant reduction in itching. Within 5
days healthy skin with normal barrier function was emerging, and
substantially complete resolution was observed in 7 days. The
administration was terminated and a follow-up on all subjects
showed complete restoration of normal skin with no visible
indication or previous disorder. Similar results were observed upon
application to skin sores, and abnormal skin areas of a number of
domestic pets.
Example 12
Data Showing how Maintaining a Healthy Digestive Tract in Animals
Promotes Growth, Reduces the Mortality Rate, and Improves the
General Health of the Animals
[0213] The binding systems interact with animal digestive mucosa to
promote healthy digestive function; provide prophylactic effect
against intestinal infection; reduce incidence and duration or
scour, improve fecal scores; reduce mortality rate; improve weight
gain rate and feed/gain ratio; improve vigor; reduce fecal shedding
of pathogens; and, reduce the effect of endotoxins. The binding
system can be used as an alternative to animal production
antibiotics, and particularly feed additives. The binding system
has a method of action distinct from current antibiotics, making it
useful against antibiotic resistant bacteria and unlikely to
promote antibiotic resistance.
[0214] The effect of the binding systems on damaged gut tissue is
to reduce irritation and inflammatory stimuli while providing
protection against further assaults until compromised tissue is
healed. As such, the use of the binding systems is an efficient
strategy for improving feed conversion ratios without the use of
antibiotics. A healthy digestive tract remains available for
maximum nutritional uptake. In comparison, appetite and immune
system stimulating additives can be counterproductive to feed
conversion maximization. Moreover, the convention wisdom is that
the use of tannin compounds in effective quantities in animal feeds
is counter-nutritional. The following represents surprising results
to those of ordinary skill in the art.
[0215] FIGS. 1A and 1B illustrate the surprising results of adding
the binding system to the drinking water of piglets, according to
some embodiments. In FIG. 1A, a binding system of green tea 50/50
extract/pomegranate in a 1:10 ratio hydrogen peroxide was
introduced in drinking water to weaned piglets to achieve a target
dosage of 2 ug total dry-plant weight equivalent per kg animal
weight). After 5 weeks, the supplemented animals cumulatively
gained 26% more weight during the period than controls. FIG. 1B
shows 93 pre-weaned piglets receiving the same target dosage in
drinking water provides reduced mortality by over 40% and improved
stool scores.
[0216] Other experiments were performed on other animals to see if
the results would be obtained in a different species. Several
hundred free range chickens were fed antibiotic free diets that
were supplemented with a similar relative quantity of the binding
system. The supplement reduced the variation in individual animal
weight, improved stool consistency, and again reduced mortality
over a control flock.
Example 13
Data Showing In Vitro Microbiologic Performance
[0217] FIG. 2 shows the minimal inhibitory concentration (MIC)
tests for a composition of 50/50 pomegranate-green tea extract
binding system with hydrogen peroxide at a ratio of 10:1 for the
hydrogen peroxide:plant compound (molar wt/dry wt) compared to the
MIC for other common antimicrobial compounds taken from published
data, according to some embodiments. The binding system has very
strong antimicrobial activity, having MIC levels similar to the
most potent of industrial biocides (Kathon). Moreover, the
performance of the binding system is notably consistent against the
gram positive and the gram negative bacteria. It's worthy to note
that all of the compounds have very different chemistry and modes
of action. All are relatively slow acting bacteriostatic compounds,
and it's important to emphasize that only RIFAXAMIN and the binding
system are intended for human consumption. The MIC range of the
binding system is also significantly times lower than MIC for
hydrogen peroxide alone.
[0218] The binding system is a 50/50 pomegranate-green tea binding
system with hydrogen peroxide at a ratio of 10:1 for the hydrogen
peroxide:phenolic compound (dry wt/dry wt). The binding system has
very strong antimicrobial activity, having MIC levels similar to
the most potent of industrial biocides (Kathon). Moreover, the
performance of the binding system is notably consistent against the
gram positive and the gram negative bacteria. It's worthy to note
that all of the compounds have very different chemistry and modes
of action. All are relatively slow acting bacteriostatic compounds,
and it's important to emphasize that only RIFAXAMIN and the binding
system are intended for human consumption. RIFAXAMIN performed
poorly compared to the binding system.
[0219] FIG. 3 shows the binding system's the effective inhibition
of a broad spectrum of bacteria by the binding system, according to
some embodiments. The binding system of FIG. 2 was used in this
example, and the selection of bacteria represent different classes
of pathogens including gram positive and gram negative types.
Similar results were obtained with several different formulations
using green tea extract, pomegranate extract and combinations
thereof. The system showed that 3-23 ug/ml of plant extract to
water was the minimal inhibitory concentration against the entire
range of bacteria. One of skill will also appreciate that this
again shows a very low concentration is needed to be effective as
an antimicrobial. The identical performance of the binding system
between the non-resistant and resistant staphylococcus strains is
an indication that the mechanism of action is unlike that of
antibiotics. Legend: the `+` indicates visible growth in broth
culture (turbidity), the `-` indicates no growth (no turbidity),
and the MIC falls within the first `+`.
[0220] FIG. 4 shows effective reduction of virus maintaining the
host cell culture viability, according to some embodiments. The
binding system of FIG. 2 was used in this example, and this figure
indicates that the binding system is not dependent on cellular
metabolism and is able to kill a virus.
[0221] FIGS. 5A and 5B are studies showing significant elevation of
polymixin B inhibition, according to some embodiments. The binding
system of FIG. 2 was used in this example, and this figure shows
that when horseradish peroxidase is added to the binding system,
effectiveness on both lipopolysaccharide endotoxin, a common food
poisoning toxin, and the cholera exotoxin, a typical protein-based
bacterial toxin, indicating the ability to inactivate a wide range
of pathogen virulence factors responsible for tissue damage,
inflammation and other undesirable physiologic effects. As such,
this is an in vitro demonstration of the increased activation
effect of the enzymes on the binding system. It also shows the
highly effecting binding on a range of toxins, a lipopolysaccharide
(has no protein structure but, rather a glucose structure) and a
protein structure, the endotoxin.
[0222] FIGS. 6A and 6F show the rapid resolution of acute watery
diarrhea in 86 subjects, according to some embodiments. The study
is a crossover study of 86 people from ages 2 and up with acute
watery diarrhea and shows rapid reduction in duration compared to a
placebo group which received treatment 24 hours later. The time
scale is last time to watery or unformed stool.
[0223] In FIG. 6A, it can be seen that upon receiving a single
1.125 mg dose of the binding system either on the first day or
second day, the mean time to the last unformed stool was 7 hours
for the subjects. FIG. 6B through 6F show significant reduction of
various secondary symptoms in the same study as FIG. 6A. In FIG.
6B, heartburn and indigestion symptoms in patients with acute
infectious diarrhea were rapidly reduced in duration compared to a
placebo group which received treatment 24 hours later. In FIG. 6C,
nausea symptoms in patients with acute infectious diarrhea were
significantly reduced compared to a placebo group which received
treatment 24 hours later. In FIG. 6D, vomiting symptoms in patients
with acute infectious diarrhea were significantly reduced compared
to placebo group which received treatment 24 hours later. In FIG.
6E, abdominal pain in patients with acute infectious diarrhea were
significantly reduced compared to a placebo group which received
treatment 24 hours later. In FIG. 6F, bloating in patients with
acute infectious diarrhea were significantly reduced compared to a
placebo group which received treatment 24 hours later.
[0224] Although these symptoms are associated with pathogen induced
acute diarrhea, those skilled will recognize that some of these
symptoms are typical of many chronic gastrointestinal conditions
such as irritable bowel syndrome (IBS), inflammatory bowel diseases
(IBD) and gastroesophogeal reflux disease. Based on the highly
effective amelioration of such systems by the polyphenol/peroxide
binding system, it is reasonable to expect similar benefits to
those suffering these other gastrointestinal conditions.
[0225] It should be appreciated that the experimental conditions
and components provided herein are for illustration and example
only. One of skill can vary the experimental conditions and
components to suit a particular or alternate experimental design.
The experimental conditions can be in vitro or in vivo, or designed
for any subject, for example, human or non-human. For example,
animal testing can be varied to suit a desired experimental method.
cm We claim:
* * * * *