U.S. patent application number 14/747004 was filed with the patent office on 2016-06-16 for synergistic compositions and methods for potentiating anti-oxidative activity.
This patent application is currently assigned to The State of Israel, Ministry of Agriculture & Rural Development, Agricultural Research Organizat. The applicant listed for this patent is Niva Shapira, The State of Israel, Ministry of Agriculture & Rural Development, Agricultural Research Organizat. Invention is credited to Joseph KANNER, Niva SHAPIRA.
Application Number | 20160166533 14/747004 |
Document ID | / |
Family ID | 32772015 |
Filed Date | 2016-06-16 |
United States Patent
Application |
20160166533 |
Kind Code |
A1 |
SHAPIRA; Niva ; et
al. |
June 16, 2016 |
SYNERGISTIC COMPOSITIONS AND METHODS FOR POTENTIATING
ANTI-OXIDATIVE ACTIVITY
Abstract
The present invention relates to synergistic compositions for
protection against oxidative damage, consisting essentially of
antacids in conjunction with antioxidants. The invention further
relates to method for preventing oxidative damage using the
compositions of the invention.
Inventors: |
SHAPIRA; Niva; (Tel-Aviv,
IL) ; KANNER; Joseph; (Rechovot, IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Shapira; Niva
The State of Israel, Ministry of Agriculture & Rural
Development, Agricultural Research Organizat |
Tel-Aviv
Beit-Dagan |
|
IL
IL |
|
|
Assignee: |
The State of Israel, Ministry of
Agriculture & Rural Development, Agricultural Research
Organizat
Beit-Dagan
IL
|
Family ID: |
32772015 |
Appl. No.: |
14/747004 |
Filed: |
June 23, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10543022 |
Sep 14, 2006 |
9060959 |
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PCT/IL2004/000071 |
Jan 25, 2004 |
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14747004 |
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60442049 |
Jan 24, 2003 |
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Current U.S.
Class: |
514/471 |
Current CPC
Class: |
A61K 33/08 20130101;
A61K 31/00 20130101; A61K 33/10 20130101; A61K 31/341 20130101;
A61P 43/00 20180101; A61K 45/06 20130101; A61P 39/06 20180101; A61K
33/06 20130101; A61K 31/00 20130101; A61K 33/06 20130101; A61K
31/05 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 31/341 20060101
A61K031/341 |
Claims
1. A composition comprising: (a) at least one antacid; (b) at least
one antioxidant, wherein the relative amount of the at least one
antioxidant in the composition is from 30 to 95% w/w of the
composition; and, optionally, (c) at least one pharmaceutically
acceptable carrier.
2. The composition of claim 1, being identified for use in
protecting against excess gastric acidity.
3. The composition according to claim 1, wherein said at least one
antacid comprises at least one classical antacid in a dose capable
of neutralizing 100 ml of 0.009 M HCl, such that the composition is
capable of elevating the pH in a stomach by at least one pH unit,
said elevating the pH being from a pH of 2 to a pH of at least
3.
4. The composition of claim 1, wherein said at least one antacid
comprises at least one classical antacid selected from a group
consisting of sodium bicarbonate, potassium hydroxide, aluminum
carbonate, aluminum hydroxide, aluminum phosphate, aluminum
hydroxycarbonate, dihydroxy aluminum sodium carbonate, aluminum
magnesium glycinate, dihydroxy aluminum aminoacetate,
dihydroxyaluminum aminoacetic acid, calcium carbonate, calcium
phosphate, hydrated magnesium aluminate activated sulfate,
magnesium aluminate, magnesium aluminosilicates, magnesium
carbonate, magnesium glycinate, magnesium hydroxide, magnesium
oxide and magnesium trisilicate.
5. The composition of claim 1, wherein said at least one antacid
comprises at least one H2-receptor antagonist and/or proton pump
inhibitor.
6. The composition of claim 1, wherein the relative amount of said
at least one antacid in the composition is from about 5 to about
90% w/w of the total antacid and antioxidant weight.
7. The composition of claim 1, wherein said antioxidant is a
polyphenol.
8. The composition of claim 7, wherein said polyphenol is selected
from the group consisting of chalcones, phenolic acid,
anthocyanins, flavonols, flavanols, flavanones, flavanonols,
hydrolyzed tannins, proanthocyanidins, phenolamines, lignans,
lignine, betalains, stilbenes, cyclic diterpenes, monoterpenes,
sesquiterpenes, sesamolin and isoflavones.
9. The composition of claim 7, wherein said polyphenol is selected
from a group consisting of anthocyanins, flavonols, flavanols,
flavanones, flavanonols, hydrolyzed tannins, proanthocyanidins,
lignine, and stilbenes.
10. A pharmaceutical composition, comprising: (a) at least one
pharmaceutically active ingredient; (b) at least one antacid; (c)
at least one antioxidant, wherein the relative amount of the at
least one antioxidant in the composition is from 30 to 95% w/w of
the composition; and, optionally, (d) at least one pharmaceutically
acceptable carrier, diluent or stabilizer.
11. The composition according to claim 10, wherein said at least
one antacid comprises at least one classical antacid in a dose
capable of neutralizing 100 ml of 0.009 M HCl, such that the
composition is capable of elevating the pH in a stomach by at least
one pH unit, said elevating the pH being from a pH of 2 to a pH of
at least 3.
12. The composition of claim 10, wherein said at least one antacid
comprises at least one classical antacid selected from a group
consisting of sodium bicarbonate, potassium hydroxide, aluminum
carbonate, aluminum hydroxide, aluminum phosphate, aluminum
hydroxycarbonate, dihydroxy aluminum sodium carbonate, aluminum
magnesium glycinate, dihydroxy aluminum aminoacetate,
dihydroxyaluminum aminoacetic acid, calcium carbonate, calcium
phosphate, hydrated magnesium aluminate activated sulfate,
magnesium aluminate, magnesium aluminosilicates, magnesium
carbonate, magnesium glycinate, magnesium hydroxide, magnesium
oxide and magnesium trisilicate.
13. The composition of claim 10, wherein said at least one antacid
comprises said at least one antacid comprises at least one
H2-receptor antagonist and/or proton pump inhibitor.
14. The composition of claim 10, wherein the relative amount of
said at least one antacid in the composition is from about 5 to
about 90% w/w of the total antacid and antioxidant weight.
15. The composition of claim 10, wherein said antioxidant is a
polyphenol.
16. The composition of claim 15, wherein the polyphenol is selected
from the group consisting of chalcones, phenolic acid,
anthocyanins, flavonols, flavanols, flavanones, flavanonols,
hydrolyzed tannins, proanthocyanidins, phenolamines, lignans,
lignine, betalains, stilbenes, cyclic diterpenes, monoterpenes,
sesquiterpenes, sesamolin and isoflavones.
17. The composition of claim 15, wherein the polyphenol is selected
from a group consisting of anthocyanins, flavonols, flavanols,
flavanones, flavanonols, hydrolyzed tannins, proanthocyanidins,
lignine, and stilbenes.
18. A method for protecting a subject from oxidative damage, the
method comprising administering to the subject a composition
comprising: (a) at least one antacid in a dose sufficient to
elevate pH in a stomach; (b) at least one antioxidant in a dose
sufficient to decrease free radical generation in the stomach; and,
optionally (c) at least one carrier, diluent or stabilizer.
19. The method of claim 18, wherein said administering results in
inhibition of peroxidation reactions in a gastrointestinal tract of
the subject.
20. The method of claim 18, wherein said administering results in
attenuation of generation of free radicals and/or peroxides in a
gastrointestinal tract of the subject.
21. The method of claim 18, wherein said at least one antacid
comprises at least one classical antacid in a dose capable of
neutralizing 100 ml of 0.009 M HCl, such that the composition is
capable of elevating the pH in a stomach by at least one pH unit,
said elevating the pH being from a pH of 2 to a pH of at least
3.
22. The method of claim 18, wherein the composition is administered
orally.
23. The method of claim 18, wherein the antacid comprises at least
one classical antioxidant selected from the group consisting of
sodium bicarbonate, potassium hydroxide, aluminum carbonate,
aluminum hydroxide, aluminum phosphate, aluminum hydroxycarbonate,
dihydroxy aluminum sodium carbonate, aluminum magnesium glycinate,
dihydroxy aluminum aminoacetate, dihydroxyaluminum aminoacetic
acid, calcium carbonate, calcium phosphate, hydrated magnesium
aluminate activated sulfate, magnesium aluminate, magnesium
aluminosilicates, magnesium carbonate, magnesium glycinate,
magnesium hydroxide, magnesium oxide and magnesium trisilicate.
24. The method of claim 18, wherein said at least one the antacid
comprises at least one antacid comprises at least one H2-receptor
antagonist and/or proton pump inhibitor.
25. The method of claim 18, wherein the relative amount of the
antacid in the composition is from about 5 to about 90% w/w of the
total antacid and antioxidant weight.
26. The method of claim 18, wherein said antioxidant is a
polyphenol, and the relative amount of said at least one
antioxidant in the composition is from 30 to 95% w/w of the
composition.
27. The method of claim 18, wherein said antioxidant is a
polyphenol selected from the group consisting of chalcones,
phenolic acid, anthocyanins, flavonols, flavanols, flavanones,
flavanonols, hydrolyzed tannins, proanthocyanidins, phenolamines,
lignans, lignine, betalains, stilbenes, cyclic diterpenes,
monoterpenes, sesquiterpenes, sesamolin and isoflavones.
28. A method for protecting a subject from excess gastric acidity,
the method comprising administering to the subject a composition
comprising: (a) at least one antacid in a dose sufficient to
elevate pH in a stomach; (b) at least one antioxidant in a dose
sufficient to decrease free radical generation in the stomach; and,
optionally (c) at least one carrier, diluent or stabilizer.
29. The method of claim 28, wherein said at least one antacid
comprises at least one classical antacid in a dose capable of
neutralizing 100 ml of 0.009 M HCl, such that the composition is
capable of elevating the pH in a stomach by at least one pH unit,
said elevating the pH being from a pH of 2 to a pH of at least
3.
30. The method of claim 28, wherein the antacid comprises at least
one classical antioxidant selected from the group consisting of
sodium bicarbonate, potassium hydroxide, aluminum carbonate,
aluminum hydroxide, aluminum phosphate, aluminum hydroxycarbonate,
dihydroxy aluminum sodium carbonate, aluminum magnesium glycinate,
dihydroxy aluminum aminoacetate, dihydroxyaluminum aminoacetic
acid, calcium carbonate, calcium phosphate, hydrated magnesium
aluminate activated sulfate, magnesium aluminate, magnesium
aluminosilicates, magnesium carbonate, magnesium glycinate,
magnesium hydroxide, magnesium oxide and magnesium trisilicate.
31. The method of claim 28, wherein said at least one antacid
comprises at least one H2-receptor antagonist and/or proton pump
inhibitor.
32. The method of claim 28, wherein the relative amount of the
antacid in the composition is from about 5 to about 90% w/w of the
total antacid and antioxidant weight.
33. The method of claim 28, wherein said antioxidant is a
polyphenol, and the relative amount of said at least one
antioxidant in the composition is from 30 to 95% w/w of the
composition.
34. The method of claim 28, wherein said antioxidant is a
polyphenol selected from the group consisting of chalcones,
phenolic acid, anthocyanins, flavonols, flavanols, flavanones,
flavanonols, hydrolyzed tannins, proanthocyanidins, phenolamines,
lignans, lignine, betalains, stilbenes, cyclic diterpenes,
monoterpenes, sesquiterpenes, sesamolin and isoflavones.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 10/543,022 filed on Sep. 14, 2006, which is a
National Phase of PCT Patent Application No. PCT/IL2004/000071,
having international filing date of Jan. 25, 2004, which claims the
benefit of priority of U.S. Provisional Patent Application No.
60/442,049 filed on Jan. 24, 2003. The contents of the above
applications are all incorporated by reference as if fully set
forth herein in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to compositions for protection
against oxidative damage and for reducing generation of free
radicals and peroxides, consisting essentially of antacids in
conjunction with antioxidants. The invention further relates to
method for preventing oxidative damage using the compositions of
the invention.
BACKGROUND OF THE INVENTION
[0003] Gastric acidity is necessary for food digestion and
protection against pathogenic bacteria, however, over acidity is
also known as a risk factor for stomach ulceration and stomach and
esophagus cancer. Free radicals are associated with induction of
gastric mucosal injury and stomach ulceration. A recent publication
by one of the inventors of the present invention disclosed that the
formation of radicals through lipid peroxidation is inhibited by
polyphenols in a dose-dependent manner and that polyphenols
stimulate the breakdown of hydroperoxides (Kanner et al., Free
Radic. Biol. 11:1388, 2001). Various classes of drugs are useful
for preventing excess acidity in the stomach, and elevating gastric
pH, including classical antacids comprising buffering agents,
proton pump inhibitors and histamine antagonists. U.S. Pat. No.
6,294,586 discloses methods and compositions utilizing
hydroxyomeprazole for the treatment of ulcers in humans.
Hydroxyomeprazole is also useful for the treatment of
gastroesophageal reflux and other conditions related to gastric
hyper secretion such as Zollinger-Ellison Syndrome. U.S. Pat. No.
6,288,081 discloses the use of quinolone- and
naphthyridonecarboxylic acid derivatives which are substituted in
position 7 by a 1-aminomethyl-2-oxa-7-azabicyclo[3.3.0]oct-7-yl
radical, for the therapy of Helicobacter pylori infections and
associated gastro duodenal disorders.
[0004] U.S. Pat. No. 6,284,265 discloses an antacid formulation
comprising antacid, antioxidant, oil and a selected nutritional
acceptable carrier. The antioxidant functions to prevent oxidation
and breakdown of the certain components, most likely the oil, prior
to consumption.
[0005] U.S. Pat. No. 4,806,354 discloses an improved health food
composition comprising B complex vitamins, prostaglandin E-1
precursor, analgesic, antacid and optionally a preservative sodium
benzoate. Compositions comprising the latter ingredient may further
comprise a primary emulsifying agent and a flavoring agent in the
absence or in the presence of a secondary emulsifying agent and an
antioxidant.
[0006] U.S. Pat. No. 4,327,076 discloses a compressed chewable
antacid tablet comprising (a) fat composition comprising edible fat
material and antioxidants; (b) a mixture of active ingredients
comprising an antacid; and (c) direct compaction tabletting aids.
The fat composition component is the essential ingredient of the
compressed chewable antacid tablet as its presence masks
chalkiness, grittiness, dryness and astringent properties of the
tablet.
[0007] Small amounts of antioxidants are added almost invariably to
pharmaceutical and other compositions for protecting the active
ingredients during storage. They are also added commonly to
preserved foods to prolong shelf life. In small quantities,
antioxidants act to prevent oxidation of the active ingredients and
to increase shelf life of these compositions as well as of
preserved foods, while not intended to alter or improve the
efficiency of the active ingredients.
[0008] There is an unmet need to provide means for potentiating
antioxidative activities thereby reducing or preventing production
of peroxides and other free radicals and attenuating or eliminating
oxidative damage.
SUMMARY OF THE INVENTION
[0009] The present invention relates to compositions containing
antioxidants as one of two classes of active ingredient therein and
antacids as the second class of active ingredient, wherein the
antioxidant and antacid of the compositions of the invention act
synergistically.
[0010] The present invention provides compositions and methods
using same for providing protection from oxidative damage, such as
the damage caused by free radicals and peroxides. The present
invention further provides compositions and methods using same for
protecting epithelial cells in the stomach and the esophagus from
oxidative damage, such as the damage caused by peroxides and free
radicals. The compositions of the present invention consist
essentially of antacids in conjunction with antioxidants and a
pharmaceutically acceptable carrier, stabilizer or diluent.
[0011] The term "antacid" as used herein is to be construed in its
most general sense and refers to materials capable of inhibiting
acid production in the stomach or attenuating gastric acidity.
These materials encompass broadly the following major categories:
classical antacids (e.g. magnesium hydroxide and aluminum
carbonate), H.sub.2-receptor antagonists or H.sub.2 antagonists
(e.g. cimetidine, ranitidine, famotidine and nizatidine) and proton
pump inhibitors (e.g. omeprazole, hydroxyomeprazole, lansoprazole,
esomeprazole, pantoprazole and rabeprazole sodium).
[0012] The present invention is based in part on the unexpected
discovery that a formulation consisting essentially of an antacid
and an antioxidant exhibits increased antioxidative activity as
compared with the antioxidative activity of a similar formulation
devoid of the antacid component. The enhancement of anti-oxidative
activity is generally at least two-fold the activity of a similar
formulation devoid of the antacid component, preferably higher.
[0013] The inventors have now surprisingly found that introduction
of a formulation consisting essentially of an antacid in
conjunction with an antioxidant to a gastric system in the presence
of pro-oxidative catalysts, converts the activity of these
catalysts from pro-oxidative to antioxidative. Without wishing to
be bound by any particular theory or mechanism of action this
conversion may be attributed to the increase in the pH of the
system that is induced by the antacid.
[0014] The compositions of the present invention are advantageous
over formulations of an antacid alone, as the protective and
antioxidative activities of the present compositions are more
effective. Thus, the compositions of the present invention may be
administered less frequently and/or at lower doses than other
antacids known in the art. The composition of the invention may
consist of a lower total amount of antacid and hence using the
composition of the invention diminishes the damaging side effects
associated with relatively high levels of antacids.
[0015] According to one embodiment, the surprising antioxidative
activity conferred in situ by the formulation of the present
invention makes these formulations particularly suitable for
preventing or attenuating the formation of gastric mucosal lesions
that are generated or exacerbated by peroxides and other flee
radicals during acid secretion and/or digestion.
[0016] As disclosed herein for the first time the strong and
effective antioxidative activity of the compositions of the present
invention is a synergistic activity per se. Neither of the active
ingredients of the formulation, namely the antacid or the
antioxidant, exerts such an efficient antioxidative activity when
administered alone, in a formulation consisting exclusively or
predominantly of this active ingredient.
[0017] According to one aspect, the present invention provides a
composition for potentiating antioxidative activities, consisting
essentially of: [0018] (a) at least one antacid component in a dose
sufficient to elevate the pH in the stomach; [0019] (b) at least
one antioxidant component in a dose sufficient to decrease free
radical generation in the stomach; and, optionally, [0020] (c) at
least one pharmaceutically acceptable carrier.
[0021] According to the present disclosure the antacid will be
present in a dose sufficient to elevate the gastric pH by at least
one pH unit, preferably the elevation will be greater than one pH
unit.
[0022] The terms "sufficient" or "sufficient dose" are
interchangeable as used herein to describe an amount that is
therapeutically effective for exerting any one of its desired
activities in vivo.
[0023] According to one embodiment, the compositions have the
ability to decrease at least two fold the concentration of free
radicals and peroxides in the stomach or esophagus as compared with
the ability of a composition comprising the same dose of
antioxidant in the absence of antacid.
[0024] According to one embodiment, the compositions of the
invention may further comprise at least one of the following
ingredients: filler, disintegrant, anticaking agent, film coating,
binder, sweetening agent, glidant, flavor, color, lubricant, and
stabilizers for solution or for solid forms, as are well known in
the art of pharmaceuticals.
[0025] According to another aspect the synergistic anti-oxidative
effects of the compositions of the present invention may be
beneficially utilized to prevent oxidation of a pharmaceutically
active ingredient during the passage of the pharmaceutically active
ingredient through the stomach or GI tract.
[0026] According to one embodiment, the present invention provides
a pharmaceutical composition, for preventing oxidation of an active
pharmaceutical ingredient in the stomach consisting essentially of:
[0027] (a) at least one pharmaceutically active ingredient; [0028]
(b) at least one antacid component in a dose sufficient to elevate
the pH in the stomach; [0029] (c) at least one antioxidant
component in a dose sufficient to decrease free radical generation
in the stomach; and [0030] (d) at least one pharmaceutically
acceptable carrier, diluent or stabilizer.
[0031] According to one embodiment, the antacid component comprises
at least one classical antacid selected from the group consisting
of: aluminum carbonate, aluminum hydroxide, aluminum phosphate,
aluminum hydroxy carbonate, dihydroxy aluminum sodium carbonate,
aluminum magnesium glycinato, dihydroxy aluminum aminoacetate,
dihydroxyaluminum aminoacetic acid, calcium carbonate, calcium
phosphate, hydrated magnesium aluminate activated sulfate,
magnesium aluminate, magnesium aluminosilicates, magnesium
carbonate, magnesium glycinate, magnesium hydroxide, magnesium
oxide and magnesium trisilicate.
[0032] According to yet another embodiment, the antacid component
comprises at least one H.sub.2-receptor antagonists selected from
the group consisting of cimetidine, ranitidine, famotidine and
nizatidine.
[0033] According to yet another embodiment, the antacid component
comprises at least one proton pump inhibitor selected from the
group consisting of: omeprazole, hydroxyomeprazole, lansoprazole,
esomeprazole, pantoprazole and rabeprazole sodium.
[0034] Some antacids may act by more than one mechanism such as the
salts of bismuth that may act both as buffers and inhibitors of
acid secretion. Appropriate salts of bismuth include bismuth
aluminate, bismuth carbonate, bismuth subcarbonate, bismuth
subgallate, bismuth subnitrate, and the like.
[0035] According to certain embodiments, the relative amount of the
antacid component in the formulation is within the range of from
about 5 to about 75% by weight and preferably from about 30 to
about 60% by weight of the total antacid and antioxidant
weight.
[0036] According to yet another embodiment, the antioxidant
component comprises one or more ingredients selected from the group
consisting of polyphenols, buffering agents (e.g. ammonium
carbonate, ammonium phosphate, boric acid, potassium citrate,
potassium metaphosphate, potassium phosphate monobasic, sodium
acetate, sodium citrate, sodium lactate solution, dibasic sodium
phosphate and monobasic sodium phosphate) reducing agents (e.g.
ascorbic acid and salts thereof, ascorbyl palmitates, thiols such
as glutathione and derivatives thereof, glucosinolates,
isothiocyanate, .alpha.-tocopherol, tocopherols and tocotrienols,
carotenoids) and plant-derived antioxidants (e.g, polyphenols such
as red wine polyphenols, plant extracts including but not limited
to grape extracts, persimmon extracts, pomegranate extracts,
bilberry extracts, blueberry extracts, rosemary extracts, sage
extracts, oregano extracts, sapodila extracts, curcum extracts,
ginger extracts, olive fruits and olive leave extracts, red beet
extracts).
[0037] According to a currently more preferred embodiment, the
antioxidant component is a polyphenol selected from the group
consisting of: chalcones (e.g. phlonidzin); phenolic acid (e.g.
caffeic acid); anthocyanimns (e.g. malvidin-3-glucoside); flavonol
(e.g. quercetin); flavanols (e.g. catechin); flavanones (e.g.
hesperitin); flavanonols (e.g. astilbin); hydrolyzed tannins (e.g.
punicalin); proanthocyanidin (e.g. epigallocatechin); phenolamine
(e.g. dopamine, L-Dopa); lignans (e.g. curcumin); lignine;
betalains (e.g. betanin); stilbenes (e.g. resveratrol); cyclic
diuterpenes (e.g. carnosolic acid, carnosol); mono and
sesquiterpenes (e.g. thymol); sesamolin (sesamol) and isoflavones
(e.g daidzein, genistein).
[0038] According to certain embodiments, the relative amount of the
antioxidant component in the formulation is within the range of
from about 30 to 95%, preferably 35 to about 65% by weight and more
preferably from about 40 to about 60% by weight of the final
formulation.
[0039] According to yet another embodiment, the at least one
pharmaceutically active ingredient is vulnerable to oxidative
damage. According to yet another embodiment, the at least one
pharmaceutically active ingredient is selected from the group
consisting of: hormones and antibiotics.
[0040] According to yet another embodiment the composition is
provided in any physical form suitable for oral administration, the
composition having a physical form selected from the group
consisting of: tablet, compressed tablet, spheroid, powder,
capsule, suspension and liquid.
[0041] According to yet another aspect the present invention
provides methods for systemic protection from oxidative damage. In
alternative embodiments there are provided methods for protecting
the GI tract from oxidative damage and further for preventing
peroxidation and generation of peroxides and free radicals in the
stomach. According to certain embodiment, the invention provides
method for potentiating antioxidative activity.
[0042] According to one embodiment, the present invention provides
a method for systemic protection from oxidative damage, comprising
administering to a subject a composition consisting essentially of
(a) at least one antacid component in a dose sufficient to elevate
the pH in the stomach; (b) at least one antioxidant component in a
dose sufficient to decrease free radical generation in the stomach;
and, optionally, (c) a carrier, diluent or stabilizer.
[0043] According to yet another embodiment, the method of the
invention is for protecting the stomach and the esophagus from
oxidative damage.
[0044] According to another embodiment, the present invention
provides a method for inhibiting peroxidation reactions in the GI
tract comprising administering to a subject a composition
consisting essentially of: (a) at least one antacid component in a
dose sufficient to elevate the pH in the stomach; (b) at least one
antioxidant component in a dose sufficient to decrease free radical
generation in the stomach; at least one carrier, diluent or
stabilizer.
[0045] According to yet another embodiment, the present invention
provides a method for attenuating generation of peroxides and free
radicals in the GI tract comprising administering to a subject a
composition consisting essentially of: (a) at least one antacid
component in a dose sufficient to elevate the pH in the stomach;
(b) at least one antioxidant component in a dose sufficient to
decrease free radical generation in the stomach; and, optionally
(c) at least one carrier, diluent or stabilizer.
[0046] The inventors previously disclosed (Kanner et al., ibid)
that peroxidation in the stomach is continued even after a meal,
under conditions that favor oxidation such as low pH and the
presence of dissolved oxygen and optionally catalysts, such as
ferrous ions. Thus, it is advantageous to apply the methods of the
invention any time during the day in order to increase the
antioxidative activity and to disintegrate free radicals and
peroxides such as lipid peroxides.
[0047] According to one embodiment, the composition is administered
to a subject before a meal or in parallel to a meal. According to
an alternative embodiment, the composition is administered to a
subject in need thereof several times a day.
[0048] According to another aspect, the principles of the present
invention may be used for improving ulcer and ulcer related
symptoms. Thus, the scope of the invention includes supplementing
administration of an antacid by parallel or consecutive
administration of an antioxidant, preferably an antioxidant which
is compatible to the antacid.
[0049] According to a preferred embodiment, the composition is
preferably administered orally as a nutraceutical consisting of
essentially non-prescription active ingredients.
[0050] Other objects, features and advantages of the present
invention will become clear from the following description and
drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0051] FIG. 1 demonstrates pH dependency of lipid peroxidation in
oxidized muscle in control samples (C) or in samples containing
quercetin (Q; 2 mM).
[0052] FIG. 2 presents .beta.-carotene co-oxidation in simulated
gastric fluid (pH2; triangle) in the presence of Fe.sup.3+ with
catechin (circle) or without catechin (square).
DETAILED DESCRIPTION OF THE INVENTION
[0053] One of the major problems addressed by the present invention
is the need to regulate gastric and esophagus acidity wherein on
one hand gastric acidity is necessary for digestion, protection
against pathogenic bacteria, stimulation of peristaltic movements
and functions of the stomach and digestive system. On the other
hand, over acidity is associated with inflammation, ulceration and
cancer of the stomach and the esophagus. The presence of
non-protected epithelial cells in the stomach and the esophagus is
another factor, which contributes to ulceration processes and the
concomitant painful symptoms and risks.
[0054] Radical production in the gastro-intestinal track is induced
by numerous factors, such as restraint stress, spicy food diet,
high-fat diet and ethanol consumption. These factors promote
superoxide anion generation, hydroxyl radical production, lipid
peroxidation, increased membrane microviscosity and increases in
DNA fragmentation in the gastric mucosa cell (GC; Bagchi D et al D.
S. Dis. Sci 1999 44:2419-28). Nevertheless, stomach acidity is in
fact facilitating and even amplifying peroxidation, which may be
further catalyzed by metmyoglobin or iron ions. The cross reactions
between lipid free radicals, generated during lipid peroxidation,
and other food constituents dramatically enriches the range of
cytotoxic and atherogenic compounds produced in the stomach such as
peroxides, hydroperoxides, oxycholesterols, malondialdehyde, and
hydroxy-alkyls. Hence, lipid peroxides are not only formed in
foods, but may also be generated during digestion, especially in
the gastric fluid, which contains absorbed oxygen and has a low pH
suggesting that the human gastric fluid is an excellent medium for
enhancing the oxidation of lipids and other dietary constituents
(Kanner at al, ibid).
[0055] The present invention provides compositions and methods
using same for potentiating antioxidative activity, for systemic
protection against oxidative damage and for protecting epithelial
cells in the GI tract from oxidative damage, such as the damage
caused by free radicals and peroxides. The compositions of the
present invention consist essentially of antacids, antioxidants and
a pharmaceutically acceptable carrier, stabilizer or diluent.
[0056] Typically, antioxidants are added to pharmaceutical and
nutritional compositions comprising antacids, in order to protect
one or more of the active ingredients from oxidative damage. Such
protection is commonly achieved using relatively low levels of
antioxidants, in the range of a few percentages by weight of the
final formulation. However, the compositions of the present
invention comprise up to 50% antioxidants or even more for the
purpose of protecting the stomach and esophagus from oxidative
damage. Thus, in the presence of at least one additional
pharmaceutical active ingredient or nutritional ingredient to the
formulation of the invention the additional ingredient may be also
protected from oxidative damage as a `by-product` of the overall
antioxidative activity.
[0057] Moreover, antioxidant potency with regard to its
antioxidative activity is affected by environmental conditions.
Under certain condition an antioxidant may function as a
pro-oxidant. Specifically, an antioxidant is in fact more potent in
mild acidic condition than in strong acidic conditions. Thus, for
preserving the effective and valuable antioxidative activity of
antioxidants in the stomach, such molecules should be protected
from the acidic environment in the stomach. The formulations and
method of the present invention provide a practical solution to
this problem, by essentially consisting an antioxidant together
with antacid.
[0058] The term "pharmaceutically acceptable carrier" refers to a
carrier medium that does not interfere with the effectiveness of
the biological activity of the active ingredient, is chemically
inert and is not toxic to the patient to whom it is
administered.
[0059] The present invention provides a formulation consisting
essentially of antacid and an antioxidant component which generally
exhibits at least two-fold of the antioxidative activity as
compared with the antioxidative activity of a similar formulation
devoid of the antacid component, suggesting synergistic connection
between these two components.
[0060] The term "synergistic" as used herein refers to a
combination which is more effective than the additive effects of
any two or more single agents. A determination of a synergistic
interaction between an antacid and an antioxidant such as a
polyphenol, may be based on the results obtained from the ferrous
ion oxidation-xylenol orange (FOX2) method as described in the
Example section, infra. The results of these assays are analyzed
using spectral analysis at 560 am with H.sub.2O.sub.2 standard
curve.
[0061] Another major problem addressed by the present invention is
the two contradicting activities that are associated by
antioxidants and pro-oxidation catalysts. The antioxidative
activity of an antioxidant may be reversed to a pro-oxidative
activity under certain environmental conditions. Antioxidants and
pro-oxidation catalysts act as antioxidants in mild acidic
condition rather than in strong acidic conditions. Thus, elevating
the pH in the stomach by using antacids together with antioxidants,
according to the present invention, enables manifestation of the
desired pro-oxidative activity. In that sense, the synergism
between the antacid and antioxidant component of the formulation of
the invention may be interpreted as the protective role of the
first component on the antioxidative activity of the latter
component. The low pH in the stomach reduces the potent
antioxidative activity of an antioxidant and further encourages
oxidation. The addition of antacids to the acidic environment, in
accordance with the principles of the invention, reduces the rates
of oxidation in the stomach by elevating the antioxidative activity
of the antioxidant.
[0062] An unexpected phenomenon established by the inventors of the
present invention is that that in the stomach, generation or
breakdown of peroxides by endogenous catalysts or catalysts
obtained from food products, or components thereof, and food
supplements can be selected by polyphenols and antacids. In other
words, the appropriate combination of polyphenol and antacids can
invert the catalyzed reaction from pro-oxidation to antioxidation
consequently attenuating lipid peroxidation in the stomach. This
principle was observed in a simulated gastric system comprising
catechin or red wine polyphenols and metmyoglobin catalyst. At low
acidic conditions, which were obtained by the addition of antacid
to the system, the metmyoglobin catalyzed the breakdown of
peroxides to zero, and completely prevented lipid peroxidation and
.beta.-carotene co-oxidation, as exemplified hereinbelow.
[0063] The terms "a gastric-simulating system" or "simulated
gastric system" are interchangeably used herein to describe an
experimental setup comprising components which simulate, in-vitro,
the environment of the GI tracts or components thereto. In a
particular example, a gastric simulating system may comprise the
following components: HCl 0.1N and pepsin (Kanner et al. ibid).
[0064] The compositions of the present invention confer the
following advantages over formulations of an antacid alone: [0065]
(I) as the protective and antioxidative activities of the present
compositions are more effective, the compositions of the present
invention may be administered less frequently than other antacids
alone; [0066] (II) as the protective and antioxidative activities
of the present compositions are more effective, the compositions of
the present invention may be administered at lower doses than
antacids alone; and [0067] (III) the composition of the invention
may consist of a lower total amount of antacid and hence using the
composition of the invention diminishes the damaging side effects
associated with relatively high levels of antacids.
[0068] The notable high antioxidative activity conferred in situ by
the formulation of the present invention makes these formulations
particularly suitable for preventing gastric and esophagus mucosa
lesion and gastric or esophagus associated lesions and cancers that
are caused inter alia by generation of free radicals and
peroxides.
Antacids and Use Thereof for Gastric Acidity
[0069] Approximately 40% of the U.S. population experiences
heartburn at least once a month, and up to 7% experiences heartburn
daily. Heartburn is the most common clinical manifestation of
gastro esophageal reflux disease (GERD) in human. Most people with
GERD self-treat the disorder by avoiding foods that seems to
aggravate symptoms, life-style alteration and acid suppressors
medications such as classical antacid, proton pump inhibitors and
acid-suppressor Hz-receptor antagonists.
[0070] Physiologically, gastric acidity is necessary for food
digestion and protection against pathogenic bacteria and even
cancer. Thus, decreasing gastric acidity confers damaging side
effects, namely, increased risk of imbalanced gastric flora
(increased Helicobacter pylori), inflammation, and cancer of the
stomach and esophagus.
[0071] However, over acidity is also known as a risk factor for
stomach ulceration and stomach esophagus cancer. Common methods for
reducing gastric acidity involve using agents that are capable of
increasing the pH of the stomach, also termed herein "antacid".
Surgical approaches for treating severe heartburn were found less
effective than oral administration of antacid.
[0072] Clinical conditions typically regulated by antacids are:
[0073] 1) Heartburn: Heartburn is commonly described as burning
feeling in the chest after a meal or when reclining. It may also
appear in the course of exercise. The cause of pain evolves from a
back flux of the stomach acid fluids into the esophagus. During
normal food consumption food travels from the mouth down the
esophagus through a one-way valve, the lower esophageal sphincter
(LES) which is the opening to the stomach. It normally opens only
upon swallowing, thus enabling food to enter the stomach, and then
it quickly closes.
[0074] In the event of access acidity or if the LES in not
functioning properly, food and stomach acidic fluids may reflux
back into the esophagus. This fluid reflux irritates the lining
cells of the esophagus and causes heartburn. In cases where the
stomach cannot prevent such reflux, the heartburn may worsen.
[0075] 2) Stomach ulcer: A stomach gastric ulcer is a raw area or
open sore that develops in the layer of cells lining the stomach.
Another layer of cells that produce mucus normally protects the
lining layer. The mucus prevents the stomach from being injured by
stomach acidic fluids. When the protective layer is damaged, an
ulcer may be generated.
[0076] Stomach ulcer is a major problem in racehorses and foals.
Studies in England, Ireland, Hong Kong and the United States report
that 80-90% of racehorses in training, 84% of yearlings and 51% of
foals have stomach ulcers. Other research found that 50% of ponies
on concentrates had ulcers, whereas ponies on hay diets did not. In
addition, 30% of death incidences in foals between 1 and 4 months
of age, are related to stomach ulcers. The period of highest risk
for developing ulcers is from 2 days to 8 weeks after birth,
especially in foals with diarrhea. The two major causes of gastric
ulceration in racehorses and foals are: (1) reduced ability of the
stomach to defend itself against gastric acid and digestive
enzymes. Stress reduces normal protective mechanisms within the
stomach. The major risk period is between 1 and 3 months of age,
that is, before foals have begun eating sufficient solid food for
saliva production to buffer stomach acid production, and (2)
Prolonged exposure of the stomach to high acid levels. High acid
levels result from modern feeding practices. Both fasting and high
grain diets predispose to gastric ulceration. Horses and ponies in
stables and those on high raw grain diets have more ulcers than
those at pasture or on hay diets.
[0077] 3) GERD: Gastroesophageal reflux disease (GERD). This
clinical condition is not only unpleasant, but may also put a
subject in a risk for Barrett's esophagus which is strongly related
to incidences of adenocarcinoma of the esophagus. Heartburn is the
most common clinical manifestation of GERD. For most people, GERD
is more of a nuisance than a threat, as in the majority of the
patients suffering from GERD ulcer, together with the subsequent
risk of bleeding or stricture formation, is not developed in the
esophagus. Other ulcer-associated problem are related to the
upper-airways, including asthma, aspiration pneumonia, or chronic
cough. Only 5% to 10% of patients with complaints of reflux have
demonstrable endoscopic esophagitis.
[0078] 4) Injury of esophageal epithelial cells: injury to
esophagus as opposed to that of stomach and duodenum is generated
due to one or more of the following factors: (i) lack of mucus and
bicarbonate secretion by surface epithelial cells; (ii) lack of
defensive enhancement by prostaglandin release; (iii) lack of an
effective mucus cap after injury; and (iv) lack of capacity to
rapidly heal erosions by the process of epithelial restitution.
[0079] 5) Gastro-esophageal cancer: It is common knowledge that
subjects suffering from chronic heartburn and acid reflux are prone
to develop esophagus cancer, including adenocarcinoma, the most
lethal form of esophageal cancer. Without limitation, it is assumed
that because the esophagus is not protected with a lining
protective layer of cells, like the stomach, stomach acid fluids
irritates the lower esophagus and, over time, trigger pre-cancerous
changes in this site that may progress into cancer. Whereas the
overall, risk for esophageal cancer is still very low (about 12,500
per year as compared with 20 to 30% of the U.S. population
reporting heartburn at least once monthly), it grows among white
men. Barrett esophagus which as mentioned above is a complication
of GERD is strongly correlated with esophageal cancer.
[0080] The term "antacid" as used herein refers to an agent which
is capable of reducing gastric acidity with the proviso that this
agent is approved for clinical or veterinary use. Antacid are
typically orally administered alkaline drugs. Antacids are capable
of raising gastric pH. In certain embodiments, antacids are capable
of raising gastric pH 4.0 or even above pH 4.0. The duration of
action of soluble antacids is usually longer than that of insoluble
antacids.
[0081] It is suggested that classical antacid exerts its activity
through the binding of the antacid cation with the chloride anions
of HC in the stomach, to form a salt. It is suggested that this
mechanism leads to partial neutralization of gastric hydrochloric
acid and inhibition of the proteolytic enzyme, pepsin. Each cation
salt has its own pharmacological characteristics that are important
for determination of which product can be used for certain
indications.
[0082] Classical antacids may be divided into families, as follows:
[0083] (a) Aluminum hydroxide: non-systemic, considerably weak and
raises pH of the stomach to 4-5 pH units. Also known to exhibit
delays gastric emptying rate and may cause constipation. [0084] (b)
Calcium carbonate (e.g. Tums.RTM.): non-systemic, may raise the pH
of the stomach to 7.0, encourages gastric emptying. May cause more
"acid rebound" than other antacids. [0085] (c) Magnesium hydroxide;
non-systemic, used as an antacid and cathartic, raises pH of
stomach contents to 9, increases gastric emptying rate. Should not
be taken by a subject suffering from renal difficulties. [0086] (d)
Sodium bicarbonate (e.g. Alka Seltzer.RTM.): systemic potent
antacid that increases gastric emptying rate. This family of
antacids may not be recommended for co-administration with other
drugs due to its systemic effect. [0087] (e) Natural antacids (e.g.
Gastro-AD.TM., Prelief.RTM.): products obtained by fermentation of
bacteria which are typically added to the food or consumed together
with food. [0088] (f) Tamer combination of the same natural active
ingredients including calcium carbonate, potassium and magnesium
hydroxide are devoid of aluminum, sodium, gelatin, and the like and
are more potent than common antacids, such as Tums.RTM.,
Maalox.RTM., and Mylanta.RTM..
[0089] An important group of antacids that may be used according to
the principles of the present invention is the group of proton pump
inhibitors. In contrast to the activity of classical antacids,
proton pump inhibitors work by completely blocking the production
of stomach acid by inhibiting (shutting down) a system in the
stomach known as the proton pump. This system is also known as
`hydrogen-potassium adenosine triphosphate enzyme system`. Proton
pump inhibitors are used to heal stomach and duodenal ulcers. This
includes stomach ulcers caused by taking nonsteroidal
anti-inflammatory drugs. They are also used to relieve symptoms of
oesophagitis (inflammation of the oesophagus or gullet) and severe
gastro-oesophageal reflux, a condition where acid leaks up from the
stomach into the gullet.
[0090] Substituted benzimidazoles are substituted sulfoxides which
are potent inhibitors of gastric acid secretion. Such substituted
sulfoxides are described for example in European Patent No. 0005129
B1. These compounds are susceptible to degradation and/or
transformation in both acid and neutral media. The acidic
decomposition of these acid labile compounds is due to an acid
catalyzed reaction described by G. Rackur et al., in Biochem.
Biophys. Res. Commun. 1985: 128(1). P477-484.
[0091] In combination with certain antibiotics (e.g, amoxycillin
and clarithromycin), proton pump inhibitors are used for avoiding
Helicobacter pylori infection.
[0092] Proton pump inhibitors are also the drugs of first choice
for a rare condition called Zollinger-Ellison syndrome. This is a
condition where a tumor in the pancreas causes too much stomach
acid to be produced and so leads to severe stomach ulceration.
[0093] Treatment with proton pump inhibitors may be involved with
several side effects. The most common side effects are diarrhoea,
nausea, constipation, wind, abdominal pain and headaches. Very
rarely they can also cause allergic reactions, itching, dizziness,
swollen ankles, muscle and joint pain, blurred vision, depression
and a dry mouth. A problem that can occur with long-term use of
proton pump inhibitors is stomach infections. Stomach acid helps to
kill microscopic organisms (microbes) such as bacteria in the
stomach. Because proton pump inhibitors completely stop acid
production using them can lead to a growth of microbes in the
stomach.
[0094] Use of proton pump inhibitors may have to be strictly
monitored in subjects suffering from liver or kidney problems, or
that are pregnant or breastfeeding. Additionally, it is not
recommended to use proton pump inhibitors in conjunction with other
medicines or herbal remedies. For example, the effects of phenytoin
(an epilepsy medicine) and warfarin (for preventing blood clots)
are affected by some of the proton pump inhibitors. Furthermore,
the absorption of the antifungal drugs ketoconazole and
itraconazole are reduced by proton pump inhibitors and the
breakdown of diazepam in the body may be blocked by some of the
proton pump inhibitors so that there is an increased effect of
diazepam.
[0095] Yet another group of antacids that may be used in the
context of the present invention is that of "H.sub.2-receptor
antagonists" or "H.sub.2 antagonists". The activity of these
compounds is related to histamine signaling. Histamine is a
naturally produced molecule that stimulates certain cells in the
stomach to produce acid by attaching, or binding, to their H.sub.2
receptors. As a result of Histamine binding to the H.sub.2
receptors cellular acid production is enhanced. The H.sub.2
antagonists function by binding to the H.sub.2 receptors without
triggering acid production.
[0096] The H.sub.2 antagonists are taken to heal stomach and
duodenal ulcers including ulcers caused by taking a nonsteroidal
anti-inflammatory drug. In order to avoid ulcer-reoccurrence
treatment with H.sub.2 antagonists may be extended to long-term
periods in a reduced dose. The H.sub.2 antagonists can also be used
to relieve indigestion, heartburn and gastro-oesophageal reflux
(GERD), a condition caused by acid leaking up from the stomach into
the bottom of the gullet. One of the H.sub.2 antagonists is also
available in a form that contains bismuth citrate (ranitidine
bismuth citrate). This is used, along with other drugs, to avoid
Helicobacter pylori infection.
[0097] The most common side effects associated with treatments with
H.sub.2 antagonists are diarrhoea and other digestive disturbances,
headache, dizziness and tiredness, and hair loss with cimetidine
and sweating with nizatidine.
[0098] Cimetidine should be avoided in people taking on warfarin,
phenytoin and other drugs for epilepsy, theophylline or
aminophylline, and some drugs for abnormal heart rhythms because it
can affect the way the body breaks them down.
[0099] Antacid treatment is usually given via oral administration
using orally suitable forms such as tablets or syrups.
[0100] Mild incidences of heartburn respond well to antacid
medications, by relieved heartburn and decrease in the associated
pain and nuisance of reflux esophagitis. Relief with liquid
antacids is most often achieved faster with the corresponding solid
alternatives, such as a tablet. Antacids have been used for
duodenal and gastric ulcers, stress gastritis, gastro-esophageal
reflux disease, pancreatic insufficiency, non-ulcer dyspepsia, bile
acid mediated diarrhea, bile mediated reflux, constipation,
osteoporosis, urinary alkalization and chronic renal failure as a
dietary phosphate binder. Histamine H.sub.2-receptor antagonists
and proton pump inhibitors has significantly reduced usage of
classical antacids for duodenal and gastric ulcers and
gastro-esophageal reflux disease.
[0101] However, antacids can still be useful for stress gastritis
and non-ulcer dyspepsia. Antacids are likely to continue to be used
for non-ulcer dyspepsia, minor episodes of heartburn
(gastro-esophageal reflux disease) among other similar
indications.
[0102] Antacids may also be given in combination with simethicone
(also known as tradenames: Gas-X.RTM., Mylicon.RTM., Phazyme.RTM.
and SonoRX.RTM.), which may relieve the symptoms of flatulence, and
also to treat stomach or duodenal ulcers. With larger doses
magnesium hydroxide (magnesia) and magnesium oxide antacids produce
a laxative effect. Some antacids, e.g. aluminum carbonate and
aluminum hydroxide, are used with a low-phosphate diet to treat
hyperphosphatemia, and can also be used with a low-phosphate diet
to prevent the formation of renal calculi.
[0103] Although most adverse effects from antacids are minor with
periodic use of small amounts, when large doses are taken for long
periods of time, significant adverse effects may occur (see for
example, Maton et al, Drugs 1999, 57:855-70). Several disadvantages
are associated with use of antacids:
[0104] A. Acid rebound. All antacids cause acid "rebound". Although
the use of antacids contributes to immediate relief of the over
acidity sensation, the duration of the effect is sometimes limited
and only relieves the acidity in the lower esophagus, where the
sensitive lining tissues in the stomach are not protected by the
treatment. As a result, antacid treatment is extended to long time
periods while not providing the required complete healing effect.
This may result in acidity "rebound" intensive use of antacids.
[0105] The terms "rebound" or "acid rebound" as used herein
interchangeably in reference to the limited effectiveness of
various antacids in reducing gastric acidity.
[0106] B. Cost effectiveness. Intensive non-efficient use of
antacid is costly as it involves medical appointments, the costs of
prescription and medications and indirect costs such as absence
from work.
[0107] C. Cross-drug interaction. Antacid interaction with other
drugs is well documented, though it may be avoided by rescheduling
administration times. However, rescheduling administration times
can be inconvenient and may discourage compliance of the complete
treatment with all medications. For example, fluoroquinolones
potency is reduced when co-administered with antacid.
Co-administration of cellulose sodium phosphate together with
calcium-containing antacids may reduce the potency of cellulose
sodium phosphate. Magnesium-containing antacids may also prevent
effective drug activity. Aluminum-containing antacids may reduce
the effects of tetracyclines and isoniazid antibiotics (also known
as INH.RTM., Laniazid.RTM. and Nydrazid.RTM.). Antacids may also
decrease the activity of Ketoconazole, Mecamylamine and
Methenamine. Sodium polystyrene sulfonate resin (SPSR) may decrease
the effectiveness of antacids.
[0108] D. Promoting the activity of cancer-related bacteria.
Antacid treatments was shown to correlate with a growth of
Helicobacter pylori ulcer bacteria, which is known to be associated
with cancer. It is speculated that the lower acidity produced by
the antacid promotes the growth of the bacteria. Helicobacter
pylori (H. pylori) has been identified in the pathogenesis of
chronic active gastritis and peptic ulcer disease and is
epidemiologically associated with gastric cancer and lymphoma. It
has been shown that production of reactive oxygen species (ROS) in
cultured gastric adenocarcinoma cells (ATCC CRL/1739) is enhanced
by at least 3.5 folds in the presence with H. pylori and in
comparison to a similar culture devoid of H. pylori. H. pylori was
also associated with DNA fragmentation in gastric mucosal cells.
The cytotoxin 87-kDa rich-H. pylori strain 60190 induced greater
production of ROS and DNA fragmentation in mucosal cells as
compared to the supernatant preparation from H. pylori strain
60190-v1, in which the cytotoxin gene has been disrupted suggesting
that H. pylori damage is related to the cytotoxin induced ROS (see
for example, Dig. Dis. Sci. 2002, 47:1405-12).
[0109] E. Powerful antacids might increase cancer risks. The
gastrin hormone in the stomach stimulates the production of gastric
acid. Certain conditions that increase the gastrin hormone in the
stomach also increase the risk of stomach cancer. Previous research
has shown that strong acid-reducing medications increase the level
of gastrin in the stomach.
[0110] F. Increased inflammatory response. It was found that
extensive treatment with several antacids elevates significantly
the bacteria populations and causes the development of stomach
inflammation. This phenomenon may be however eliminated by
co-administration of antibiotics with antacids.
Protection of the Gastric System by Antioxidants and Antioxidative
Mechanisms
[0111] Free radical production occurs continuously in all cells as
part of normal cellular function. However, excess free radical
production originating from endogenous or exogenous sources might
play a role in many diseases.
[0112] Human gastric mucosa contains a highly active peroxidase in
addition to the myeloperoxidase contributed by neutrophils. The
activity of endogenous peroxidase, a major H.sub.2O.sub.2
metabolizing enzyme, was recently reported showing its critical
role in controlling oxidative damage in gastric mucosa. When
myeloperoxidase level increases due to neutrophil accumulation,
gastric peroxidase (GPO) level decreases significantly. This leads
to further accumulation of endogenous H.sub.2O.sub.2 that can cause
more oxidative damage and aggravate the ulcer. Also mucosal total
superoxide dismutase (Mn and Cu--Zn SOD) level decreases
significantly under accumulation of reactive oxygen metabolites
(ROM), leading to increased accumulation of O.sub.2. Gastric ulcer
is associated with oxidative damage of the mucosa as evidenced by
significant increase in lipid peroxidation, protein oxidation, and
thiol depletion indicating accumulation of ROM. It is also
recognized that many other diseases are caused or provoked by a
free radical oxidation mechanism e.g. cancer, cataracts and
diabetes among others.
[0113] Potent antioxidants are capable of attenuating and even
preventing the formation of radicals by scavenging them, promoting
their decomposition and reversing the harmful actions of reactive
oxygen and nitrogen species, which are generated in vivo. One of
the major damaging effect of reactive oxygen and nitrogen species
is destruction of DNA, lipids, proteins, and other biomolecules.
Endogenous antioxidant defenses, such as superoxide dismutases,
H.sub.2O.sub.2-removing enzymes and metal binding proteins, are
inadequate for a complete prevention of the damage associated with
the reactive species, and thus an exogenous supply of antioxidants
is important. Many dietary compounds have been suggested to be
important antioxidants, including vitamins E and C and especially
flavonoids. Some antioxidants can exert pro-oxidant effects in
vitro, but their physiological relevance is uncertain.
[0114] In considering the biological importance of dietary
antioxidants, attention was usually focused on those that are
absorbed through the gastrointestinal (G) tract into the rest of
the body. Recent publications raised the argument that the high
levels of antioxidants present in certain foods (fruits,
vegetables, grains) and beverages (e.g. green tea) play an
important role in protecting the gastrointestinal tract itself from
oxidative damage, and in delaying the development of stomach, colon
and rectal cancer. Carotenoids and flavonoids do not seem to be as
well absorbed as vitamins C and E. Hence their concentrations can
be much higher in the lumen of the GI tract than in the plasma or
other body tissues, enabling their antioxidative action in the GI
tract. Additional protective mechanisms of these dietary
constituents (e.g. effects on intercellular communication,
apoptosis, cyclooxygenases and telomerase) may also be
important.
[0115] Polyphenols is a class of substituted phenolic compounds
that are also known as flavanols or catechins and are commonly
found in plant extracts. Polyphenols occur abundantly in red wine
and consist of a large number of different chemical substances of
varying molecular weights. The chief polyphenol components of
grapes and wine, and their concentrations. See for example U.S.
Pat. No. 6,642,277. Among the polyphenols are the following
classes: flavonoids (a term often used to denote polyphenols in
general, but more commonly in Europe to denote only the flavones),
the flavanols, proanthocyanidins (also called procyanidols,
procyanins, procyanidins and tannins) and anthocyanins. Within the
general term "polyphenols" are included the dihydroxy- or
tri-hydroxy benzoic acids and the phytoalexins, a typical example
of which is resveratrol.
[0116] Polyphenols may conveniently be obtained from red wine,
grape juice, or other polyphenol-containing liquids by absorption
onto a chromatographic resin column, with elution of the
polyphenol-enriched fraction from the column (typically following a
washing step) by use of a 40-80% ethanol eluent, or other suitable
organic solvent (such as methanol, acetone, ethyl acetate,
dimethylene chloride, and chloroform, which may be in aqueous
solution). The organic solvent is preferably relatively volatile
(i.e. having a boiling point of between 30 and 85.degree. C. at 760
mm Hg pressure) and so readily driven off, to leave a substantially
dry (i.e. less than 10% w/w H.sub.2O) solid composition comprising
polyphenols. Such a method may successfully be used to obtain a
total polyphenol pool from red wine.
[0117] Alternatively, polyphenols may be obtained from red wine,
grape juice, or other polyphenol-containing liquid by solvent
extraction using a suitable organic solvent immiscible with the
wine or other liquid. Alternatively, polyphenols may be obtained
from polyphenol-containing solids by solvent extraction (typically
extraction with an organic solvent such as ethanol or ethyl
acetate) the solids can then be separated from the solvent by
filtration or centrifugation. The solvent may then be evaporated to
leave a substantially dry, solid composition comprising
polyphenols.
[0118] A number of compositions are publicly available which are
prepared from wine or grape by-products and which allegedly contain
polyphenols (albeit at quite low levels in some of the
compositions). Among them is French Paradox.TM. made by preparing
an extract from marc (the grape skin waste remaining after wine
fermentation). Most of the polyphenols present in the grape skins
are alcohol-soluble, and so tend to be extracted into the
fermenting wine. Hence, French Paradox.TM. capsules have actually
rather low polyphenol content. Other publicly available
compositions include an anthocyanin-containing powder (obtainable
from Sefcal.TM.) made from a grape skin extract, and which is used
as a food colorant, and a pro-anthocyanidin-containing composition
(Endotelon.TM.) prepared from grape seeds.
[0119] Quercetin is a naturally occurring bioflavonoid with strong
antioxidant activity. The antioxidant activity of quercetin
protects the gastrointestinal tract in several ways: [0120] (I)
Quercetin prevents oxidation of lipids. The gastrointestinal tract
has an increased exposure to oxidative stress due in part to the
low pH therein. It is important to protect the lipid bilayer of the
cell wall of the gastrointestinal, tract as these cells are an
important component of the immune system. [0121] (II) Quercetin
prevents the depletion of glutathione in the cells of the
intestinal tract. Glutathione is a co-substrate for the antioxidant
enzymes glutathione peroxidase and glutathione reductase. By
preserving the accumulation of glutathione, quercetin protects
metabolic activity and cellular structure of these highly
vulnerable cells from the damage caused by toxic free radical.
Vitamin C synergistically improves the ability of the quercetin to
preserve glutathione. [0122] (III) Quercetin increases mucus
secretion from gastric cells. The mucus polysaccharide provides a
protective buffering layer for the gastric cells from the low pH of
the stomach. Such reduced contact prevents or reduces the
occurrence of gastric lesions. [0123] (IV) Quercetin induces
antispasmodic activity that prevents uncontrolled peristaltic
activity such as the activity that is associated with diarrhea. The
reduced excretion of the intestinal contents provides benefit to
the gastrointestinal tract by preventing overproduction of cellular
protective materials. [0124] (V) Quercetin reduces the immune
response to allergens by inhibiting IgE-mediated release of
molecules from mast cells. Quercetin also inhibits IgG-mediated
histamine release. Furthermore, quercetin is a potent inhibitor of
lipoxygenase, which metabolizes arachidonic acid as the first step
towards pro-inflammatory arachidonic acid mechanism. These
activities of quercetin provide a wide protection of the GI tract
against gastric lesions. These activities are enhanced in the
presence of vitamin C and prevent from antigenic proteins and other
antigenic compounds from entering to the body.
[0125] Anthocyanosides, also known as Bilberry anthocyanoside, is a
family of plant-derived polyphenols. They are sometimes called
anthocyanidins. Typical examples are: cyanidin (hydroxylated at
positions 3, 5, 7, 3', 4'), delphinidin (hydroxylated at positions
3, 5, 7, 3', 4', 5') and pelargonidin (hydroxylated at positions 3,
5, 7, 3). The hydroxyl groups are usually glycosylated and/or
methoxylated (e.g. malvidin at 3', 5). Antocyanosides are used for
treating inflammation and ulcer. Administration of a Bilberry
anthocyanoside extract induces reduction of inflammation, in-vivo.
It has been suggested that this polyphenol may also reduce platelet
aggregation, probably by increasing release of prostacyclin, a
potent blood vessel dilating and platelet anti-aggregation factor.
Orally administered bilberry anthocyanosides prevents and even
cures ulcer activity in various experimental models of gastric
ulcer without affecting gastric secretion. This activity can be
attributed, at least partly, to an increase in gastric mucus.
[0126] Consumptions of polyphenols derived from green tea were
associated with reduced risk of stomach cancers and esophagus
cancers. An epidemiological study conducted in Shanghai, China
(1986) among 18,244 men of 45 to 64 years indicated that only half
of the `heavy` tea-drinkers developed cancer of the stomach or
esophagus as compared with `light` tea drinkers. Among different
kinds of tea, the green tea contains the highest levels of the
antioxidants polyphenolic molecules: catechins. Catechin was shown
to inhibit the growth of tumor cells and to protect healthy cells
from damage associated with cancer. U.S. Pat. No. 6,652,890
discloses methods and compositions of treating cancer or solid
tumors utilizing catechins. The compositions of catechins include
epigallocatechin gallate (EGCg) the major catechin in green tea,
epicatechin (EC), epicatechin gallate (ECG), epigallocatechin
(EGC).
[0127] Case-control study in Spain, on 354 cases of gastric cancer
and 354 controls, supports the hypothesis that the protective
effect of fruit and vegetables against gastric cancer could, in
part, be due to the presence of flavonoids. In addition, a review
of the scientific literature on the relationship between vegetable
and fruit consumption and risk of cancer, resulted from 206 human
epidemiological studies and 22 animal studies showed evidence for a
protective effect of high vegetable and fruit consumption against
cancers of the stomach, esophagus, lung as well as oral cavity and
pharynx, endometrium, pancreas, and colon. Substances present in
vegetables and fruit that may help protect against cancer, and
their mechanisms, were also briefly reviewed; these included
dithiolthiones, isothiocyanates, indole-3-carbinol, allium
compounds, isoflavones, protease inhibitors, saponins,
phytosterols, inositol hexaphosphate, vitamin C, D-limonene,
lutein, folic acid, beta carotene, lycopene, selenium, vitamin E,
flavonoids, and dietary fiber.
[0128] Numerous antioxidants, derived from medicinal plants, were
shown to have gastric cyto-protective activity. Analyses of the
chemical content of medical plants extracts showed that most of
these type of plants mainly contains the following molecules:
tannins, saponins, flavonoids and coumarius. A common family of
medicinal plant is the Stachytarpheta cayennensis. Extracts of
plants from this family showed notable anti-secretion and
anti-ulcer activities. Cholinergic and histaminergic stimulation of
acid secretion were similarly reduced by these extracts suggesting
inhibition of common steps in both pathways, possibly at the level
of histamine release/H2 receptor interaction, or at the proton
pump.
[0129] Among the various beneficial activities attributed to
polyphenols, inhibition of gastric H+, K(+)-ATPase by was also
observed. Gastric H+, K(+)-ATPase plays a pivotal role in the final
step of gastric acid secretion and some flavonoids were found to
inhibit the activity of this enzyme.
[0130] Several types of flavonoids and their metabolites, such as
Ponciretin, hesperatin, naringenin and diosmetin, were also capable
of presenting anti-Helicobacter pylori activity, in vitro.
[0131] Distribution pattern of flavonoids in the gastrointestinal
lumen and wall was shown to be accompanied by partial
deglycosylation that began in the stomach where at first quercetin
and later apigenin, chrysoeriol and isorhamnetin aglycones were
detected.
[0132] Antioxidants are advantageous over antacids in preventing
reflux oesophagitis. Preliminary experiments in rats having
surgically-induced reflux oesophagitis indicated that wormwood,
which has antioxidant properties, successfully managed reflux
esophagitis as compared to treatment with ranitidine (a common acid
suppressant anti-secretory agent). Treatment with wormwood resulted
with less severe blistering, lower levels of inflammation and more
extensive healing.
[0133] Another protective effect of antioxidants is their ability
to prevent gastropathy induced by non-steroidal anti-inflammatory
drugs (NSAID). NSAID treatment is known to induce, in a large
percentage of the subjects treated with these drugs these, gastric
ulcers and related complications, a condition which is also known
as "NSAID gastropathy". NSAID gastropathy differs from classic
peptic ulcer disease in many ways, and traditional peptic ulcer
therapy is largely ineffective in preventing NSAID-induced
gastropathy. The prostaglandin misoprostol has been shown to be
effective and is approved for the prevention of NSAID gastropathy.
Common NSAIDs include phenylbutazone (`bute`) and flunixin
meglumine (Banamine.TM.). NSAIDs act by interrupting the production
of prostaglandins. One particular prostaglandin, PgE2, plays an
important role in preventing gastric ulceration. PgE2 exerts its
preventive activity by reducing gastric acid production and by
increasing blood flow to the gastric epithelium.
[0134] U.S. Pat. No. 6,365,184 discloses an oral pharmaceutical
dosage form comprising an acid susceptible proton pump inhibitor
and one or more NSAIDs in a fixed formulation, wherein the proton
pump inhibitor is protected by an enteric coating layer. The
pharmaceutical dosage form is especially useful in the treatment of
gastrointestinal side-effects associated with NSAID treatment.
[0135] Yet another protective effect of antioxidants is their
ability to inhibit nitrous acid-dependent tyrosine nitration and
DNA base deamination. A vast range of plant polyphenol constituents
was shown to prevent damage mediated by acidic nitrite. The
epicatechin/gallate family of flavonols, constituents of green tea,
red wine, etc., demonstrated the most extensive inhibitory
properties against both tyrosine nitration and base deamination.
The ability of plant polyphenols to scavenge reactive nitrogen
species derived from acidic nitrite may explain their protective:
effects against gastric cancer.
Pharmaceutical Compositions and Modes of Administration
[0136] The formulations of the invention may be formulated into
pharmaceutical preparations for administration to a subject as a
medicament or as food supplements or as food fortifying
ingredients. The formulations may be specifically administered in
the context of pharmaceutical preparations for treatment of
gastric- and esophagus-associated diseases. In a preferred
embodiment, the subject is a mammal.
[0137] According to one embodiment, the present invention provides
a formulation consisting essentially of: [0138] (a) an antacid
component; [0139] (b) an antioxidant component; [0140] (c) a
pharmaceutically acceptable carrier; and, optionally, [0141] (d) at
least one active pharmaceutical ingredient.
[0142] According to another embodiment, the pharmaceutically
acceptable diluent, stabilize or carrier comprises at least one
substance selected from the group consisting of: water, organic
solvent, inorganic solvent, buffering agent, acidifying agent,
alkalizing agent and alcohol denaturant.
[0143] According to one embodiment, the composition of the
invention may further comprise at least one of the following
ingredients: filler (e.g. xylitol, mannitol), disintegrant (e.g.
crosspovidone), anticaking agent (e.g. silicon dioxide), film
coating (e.g. polymethacrylates), coating solution (e.g. denatured
ethanol), binder (e.g. bentonite, corn starch), stabilizer for or
for solid forms, entericoating polymer, sweetening agent (e.g.
aspartame), glidant (e.g. silicon dioxide), flavor, color (e.g.
titanium dioxide), lubricant and plasticizer (e.g.
triethylcitrate).
[0144] Formulations according to the present invention may comprise
any conventional carrier or adjuvant used in pharmaceuticals,
personal, care formulations and compositions or veterinary
formulations. These carriers and adjuvants include, the following:
[0145] (i) Acidifying agents. [0146] (ii) Alcohol denaturants, such
as, methyl isobutyl ketone and sucrose octacetate. [0147] (iii)
Alkalizing agents. [0148] (iv) Antifoaming agents, e.g. dimethicone
and simethicone. [0149] (v) Antimicrobial preservatives, such as,
benzalkonium chloride, benzalkonium chloride solution,
benzelthonium chloride, benzyl alcohol, butylparaben,
cetylpyridinium chloride, chlorobutanol, chlorocresol, cresol,
ethylparaben, methylparaben, phenol, phenylethyl alcohol,
propylparaban and propylparaben sodium. [0150] (vi) Plasticizers,
e.g. castor oil, diacetylated monoglycerides, diethyl phthalate,
glycerin, mono- and di-acetylated monoglycerides, polyethylene
glycol, propylene glycol, triacetin and triethyl citrate. [0151]
(vii) Solvents, for example, acetone, alcohol, diluted alcohol,
benzyl benzoate, butyl alcohol, carbon tetrachloride, chloroform,
corn oil, cottonseed oil, ethyl acetate, glycerin, hexylene glycol,
isopropyl alcohol, methyl alcohol, methylene chloride, methyl
isobutyl ketone, mineral oil, peanut oil, polyethylene glycol,
propylene carbonate, propylene glycol, sesame oil and water. [0152]
(viii) Sorbents, such as, powdered cellulose, charcoal, purified
siliceous earth or carbon dioxide sorbents (e.g. barium hydroxide
lime, soda lime). [0153] (ix) Stiffening agents, for example,
hydrogenated castor oil, cetostearyl alcohol, cetyl alcohol, cetyl
esters wax, hard fat, paraffin, polyethylene excipient, stearyl
alcohol, emulsifying wax, white wax and yellow wax. [0154] (x)
Suppository bases, e.g., cocoa butter, hard fat and polyethylene
glycol.
[0155] Compositions comprising a compound of the invention
formulated in a compatible pharmaceutical carrier may be prepared,
packaged, and labeled for treatment of the indicated disease, such
as sarcomas and carcinomas (e.g., fibrosarcoma, colon carcinoma,
gastric carcinoma, adenocarcinoma, papillary carcinoma, papillary
adenocarcinomas, epithelial carcinoma, gastro-esophageal cancer),
GERD disease, stomach or duodenal ulcer, heartburn, peptic ulcer
disease, chronic-active gastritis, Helicobacter pylori ulcer,
gastric inflammation and reflux oesophagitis.
[0156] The formulations and their physiologically acceptable
carrier may be formulated for oral administration. For oral
administration, the pharmaceutical preparation may be in liquid
form, for example, solutions, syrups or suspensions, or may be
presented as a drug product for reconstitution with water or other
suitable vehicle before use. Such liquid preparations may be
prepared by conventional means with pharmaceutically acceptable
additives such as suspending agents (e.g. sorbitol syrup, cellulose
derivatives or hydrogenated edible fats); emulsifying agents (e.g.,
lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily
esters, or fractionated vegetable oils); and preservatives (e.g.,
methyl or propyl-p-hydroxybenzoates).
[0157] The pharmaceutical compositions may take the form of, for
example, tablets or capsules prepared by conventional means with
pharmaceutically acceptable excipients such as binding agents
(e.g., pregelatinized maize starch, polyvinyl pyrrolidone or
hydroxypropyl methylcellulose); fillers (e.g., lactose,
microcrystalline cellulose or calcium hydrogen phosphate);
lubricants (e.g., talc or silica); disintegrants (e.g., potato
starch or sodium starch glycolate); or wetting agents (e.g., sodium
lauryl sulphate). The tablets may be coated by methods well-known
in the art. In a preferred embodiment, the pharmaceutical
composition may take the form of a capsule or powder to be
dissolved in a liquid for oral consumption.
[0158] The compositions of the invention may be in any edible form
or part of a food products such as a candy bar, chocolates and the
like.
[0159] Novel pharmaceutical compositions for acid labile
substances, particularly proton pump inhibitors such as
lansoprazole or omeprazole, as well as for methods of making such
are disclosed in U.S. Pat. No. 6,296,876.
[0160] According to another embodiment, the antioxidant compound is
a polyphenols, such as a catechin. Catechins may be combined with
any antacid in intimate admixture with a pharmaceutical carrier
according to conventional pharmaceutical compounding techniques.
The carrier may take a wide variety of forms depending on the form
of preparation desired for administration, e.g., oral or parenteral
(including tablets, capsules, powders, intravenous injections or
infusions). In preparing the compositions for oral dosage form any
of the usual pharmaceutical media may be employed, e.g. water,
glycols, oils, alcohols, flavoring agents, preservatives, coloring
agents, and the like; in the case of oral liquid preparations,
e.g., suspensions, solutions, elixirs, liposomes and aerosols;
starches, sugars, micro-crystalline cellulose, diluents,
granulating agents, lubricants, binders, disintegrating agents, and
the like in the case of oral solid preparations e.g., powders,
capsules, and tablets. In preparing the compositions for parenteral
dosage form, such as intravenous injection or infusion, similar
pharmaceutical media may be employed, e.g., water, glycols, oils,
buffers, sugar, preservatives and the like know to those skilled in
the art. Examples of such parenteral compositions include, but are
not limited to Dextrose 5% (w/v), normal saline or other solutions.
The total dose of the catechins and antacids may be administered in
a vial of intravenous fluid, e.g., ranging from about 0.01 to about
1000 mg per kg body weight of catechins. The volume of dilution
fluid will vary according to the total dose administered and over
the length of the period of time of administration.
[0161] Various aspects and embodiments of the present invention are
illustrated in the following examples. It will be appreciated that
modification of detail may be made without departing from the scope
of the invention.
EXAMPLES
Materials and Methods
[0162] Metmyoglobin (metMb, from horse skeletal muscle), soybean
lipoxygenase (type I-B), .beta.-carotene, linoleic acid, Tween 20,
butylated hydroxytoluene (BI-T), catechin, pepsin (A, from porcine
stomach mucosa), ferrous ammonium sulfate. Xylenol orange, and
triphenylphosphine (TTP) were obtained from Sigma Chemical Co. (St.
Louis, Mo., USA). Sodium chloride, hydrogen peroxide (30%) and L
(+)-ascorbic acid (AA) were obtained from Merck (Darmstadt,
Germany). Ferric chloride (Fe) was obtained from Riedel-de-Haen
(Hannover, Germany). Sodium borohydride was obtained from BDH (NJ,
USA). Human gastric fluid (HGF) was collected, with consent, from a
fasting woman (age 25) during regular gastric endoscopic tests, and
was kept at -80.degree. C. Small slices grilled turkey meat was
kept at -80.degree. C. Simulated gastric fluid (SGF) was freshly
prepared according to the U.S. Pharmacopoeia.
[0163] Spectroscopic measurements: The presence of hydroperoxides
was determined by means of the ferrous ion oxidation-xylenol orange
(FOX2) method (Nourooz-Zadeh, J. Methods Enzymol. 1999, 300: 58-62)
including spectral analysis at 560 nm with H.sub.2O.sub.2 standard
curve.
[0164] Hydroperoxides measurement in linoleic acid emulsions:
Catechin was dissolved in 10% ethanol in water and the reaction
tubes were incubated in triplicate in a shaking bath at 37.degree.
C. for 180 min. Samples of 25 or 50 .mu.l were taken from the
reaction tubes during the incubation and added to 475 or 950 .mu.l,
respectively, of FOX2 reagent. Before each experiment, old stock
solution of linoleic acid was mixed with fresh solution in order to
maintain an accurately consistent initial level of
hydroperoxides.
[0165] The presence of metMb, Fe, AA, wine, or catechin, at the
concentrations used in the experiments, did not interfere with the
FOX2 assay.
Example 1
Hydroperoxides Measurement in Turkey Muscle Tissue
[0166] Muscle tissue was ground with three parts of liquid,
containing SGF with red wine, or ethanol solution or water as
controls, for 1 min in a laboratory blender (Waring.RTM., CT, USA)
and adjusted to pH 3.0. The wine was diluted with 12% ethanol
solution, so that the ethanol content (final concentration 6%) was
equal in all treatments, except for the water control.
[0167] The meat-liquid mixture in each treatment was divided among
several tubes and incubated in a shaking bath at 37.degree. C. for
180 min. At five times points, the hydroperoxides in the samples
were extracted by 10-fold dilution in methanol under slow stirring
for 15 min, following paper filtration (cat. No. 1442,
Whatman.RTM., England). Samples, 100 .mu.l, of the filtered
solution were subjected to the FOX2 assay, which included TTP
reagent controls, in order to prevent potential interference from
ferric ions and the red wine pigments.
[0168] The heated red turkey meat was found to contain about 180
.mu.M hydroperoxides and malondialdehydes, a concentration which is
10-20 times higher than that in the fresh muscle (FIG. 1).
[0169] In addition to the control samples, that is samples that
were devoid of antacid, the heated red turkey muscle was ground in
the presence of an antacid that elevated the pH to 6.5. An
antioxidant (catechin or quercetin, 2 mM) was then added (FIGS. 1
and 2).
[0170] The results show rapid lipid peroxidation in the control
samples of the grilled (oxidized) meat and after an incubation of
180 min the level of the hydroperoxides rose to more than 2000
.mu.M. Addition of antacid attenuated the acidity of the samples to
pH 5.0 and further reduced lipid peroxidation (FIG. 1). This
attenuation is also associated with the reversed, i.e. oxidative,
activity of the catalyst metMb (also see, Kanner et al., ibid).
[0171] Moreover, in the sample that contained both antacid and
catechin not only that lipid peroxidation was inhibited, but the
reaction was also reversed and hydroperoxides concentration was
reduced to substantially zero (FIGS. 1 and 2).
Example 2
.beta.-Carotene Oxidation
[0172] Several studies showed that .beta.-carotene may reduce the
risk for prostate cancer. Also, epidemiological studies showed that
this molecule is inversely correlated with risk of lung cancer
although several intervention studies, among a population of
smokers, did not confirm that .beta.-carotene protects against lung
cancer.
[0173] Recent study (Kanner J, ibid) showed a dramatic increase in
.beta.-carotene oxidation under acidic conditions of the gastric
simulated fluid. Here B carotene co-oxidation was determined in
order to identify free radical damage during lipid peroxidation in
the same model system of acid gastric fluid (FIG. 2).
[0174] The addition of metmyoglobin to the system catalyzed the
bleaching of .beta.-carotene, demonstrating free radical damage to
the target molecule. However, the addition of catechin to the same
system containing metmyoglobin inhibited .beta.-carotene oxidation
by 100% (FIG. 2).
Example 3
Antacid (Zantac.RTM.) and Antioxidant Formulation in Babies
[0175] GERD may affect up to several million children. Incidence of
GERD in infants is reported to be approximately 8% to 10%, and
prevalence of reflux esophagitis is reported to be approximately
5%.
[0176] Recurrent regurgitation and/or vomiting as well as common
GERD reflux symptoms in infants, occur in as many as 50% of
newborns (0 to 3 months) and up to 67% of 4-month-old infants.
Other symptoms include abdominal discomfort, frequent hiccups, sour
burps, bad breath, excessive crying/irritability, feeding
refusal/refusal of solid foods.
[0177] Untreated GERD may further cause or exacerbate esophagitis,
esophageal strictures, dysphagia, chronic cough, apnea, anemia,
pneumonia, asthma, failure to thrive.
[0178] ZANTAC.RTM. (GlaxoSmithKline) is a histamine
H.sub.2-receptor antagonist comprising ranitidine which is approved
for the treatment of GERD in infants from the age of 1 month.
[0179] A baby receives 10 mg/kg/day of Zantac with the addition of
an ascorbyl palmitate antioxidant in a dose of 20-50 mg/day. This
combined formulation increases the efficiency of Zantac and reduce
the overall Zantac dose and length of treatment.
Example 4
Antacid and Antioxidant Formulation in Horses
[0180] The stomach of a horse comprises two different parts:
squamous epithelium and glandular epithelium wherein a distinct
margin, called the margo plicatus, separates these two parts.
[0181] Hydrochloric acid (gastric acid) and pepsin are produced in
the glandular part which also secretes factors that help to protect
the stomach, i.e. a mucus-bicarbonate layer serves to protect the
stomach lining both by preventing acid from physical contact with
the stomach surface, and by buffering gastric acid at the level of
the stomach lining.
[0182] This mucous-bicarbonate layer protects only the glandular
part of the stomach, and not the squamous part. Other protective
factors include prostaglandin E, which causes increased blood flow
in the stomach lining, increased secretion of the mucus-bicarbonate
layer, and also causes decreases in hydrochloric acid
production.
[0183] Unlike humans, horses produce gastric acid continually,
regardless of whether they are eating regularly. On average the
gastric pH is 2.0 or less. The stomach of an un-fed horse is more
acidic than usual because acid production cannot be `turned-off`.
The squamous part of the stomach is at great risk at highly acid
environment, because it not as protected as the glandular part of
the stomach.
[0184] Both yearling and adult horses consume pellet forms of food
faster than they eat traditional grain diets. The amount of
bicarbonate in saliva increases as saliva production increases and
so the longer the horse's meal lasts the acidity in the stomach is
reduced.
[0185] Acid-suppressive therapy in horses may be carried out using
drugs that inhibit acid formation. Horses at work respond less
favorably to such therapy than horse at rest. Acid-suppressive
therapy is relatively expensive, however, if the required dose is
reduced (e.g. for economic reasons) healing may not occur.
[0186] For an antacid to work well in horses it must possess
several properties: [0187] (a) easy administration, preferably by
adding to the horse's feed; [0188] (b) a small dose must neutralize
a large amount of acid and it should coat and protect the mucosa
from gastric acid irritation; [0189] (c) the antacids has to be
able to adsorb pepsin and other substances that can damage the
mucosa; [0190] (d) it should not affect fecal consistency or other
harmful side effects.
[0191] Kentucky Equine Research has worked to develop an equine
antacid that would satisfy the above criteria. Unfortunately, the
horses became ataxic and displayed signs of tying-up following high
MgO doses (which is one of the common antacids used for humans).
Aluminum based antacid raised concern regarding interference with
phosphorus absorption, but it appears that aluminum-containing
antacids can be safely fed to horses.
[0192] Using the composition of the present invention provides the
required affect and alleviates ulcer and ulcer-related symptoms
efficiently. Exemplary recommended doses are as follows: antacid
20-200 mg per 1 liter of stomach with antioxidant and antioxidant
800 mg per 1 liter of stomach and volume. The antacid is famotidine
(20-40 mg/l, omeprazole (20 mg/l) or panteprazole (40 mg/l).
[0193] The foregoing description of the specific embodiments will
so fully reveal the general nature of the invention that others
can, by applying current knowledge, readily modify and/or adapt for
various applications such specific embodiments without undue
experimentation and without departing from the generic concept,
and, therefore, such adaptations and modifications should and are
intended to be comprehended within the meaning and range of
equivalents of the disclosed embodiments. It is to be understood
that the phraseology or terminology employed herein is for the
purpose of description and not of limitation. The means, materials,
and steps for carrying out various disclosed functions may take a
variety of alternative forms without departing from the
invention.
* * * * *