U.S. patent application number 14/088442 was filed with the patent office on 2014-03-20 for composition and methods for extracting and using phytochemicals for the treatment of influenza.
This patent application is currently assigned to POM Wonderful LLC. The applicant listed for this patent is POM Wonderful LLC. Invention is credited to Harley R. Liker, Mohammad Madjid.
Application Number | 20140080776 14/088442 |
Document ID | / |
Family ID | 43124955 |
Filed Date | 2014-03-20 |
United States Patent
Application |
20140080776 |
Kind Code |
A1 |
Madjid; Mohammad ; et
al. |
March 20, 2014 |
COMPOSITION AND METHODS FOR EXTRACTING AND USING PHYTOCHEMICALS FOR
THE TREATMENT OF INFLUENZA
Abstract
Methods and compositions for treating at least one symptom of a
viral infection, said method of treatment comprising administering
to a subject with a least one symptom of a viral infection an
effective dosage unit of a composition comprising pomegranate
extract. The compositions comprise pomegranate extract comprising
polyphenols. The composition is produced by a process comprising
providing one or more pomegranate solids selected from the group
consisting of a pericarp, inner membrane and seeds, creating a
mixture comprising said one or more pomegranate solids in an
aqueous solution, heating said mixture to a temperature that
permits enzyme catalysis of said pomegranate solids, and removing
residual insoluble solid materials from said mixture to provide
said pomegranate extract. Compositions containing the extract may
be used as a food product, beverage, pharmaceutical preparations,
nutritional supplements, vitamin supplements, food additives, and
food supplements.
Inventors: |
Madjid; Mohammad; (Houston,
TX) ; Liker; Harley R.; (Beverly Hills, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
POM Wonderful LLC |
Los Angeles |
CA |
US |
|
|
Assignee: |
POM Wonderful LLC
Los Angeles
CA
|
Family ID: |
43124955 |
Appl. No.: |
14/088442 |
Filed: |
November 24, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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12730119 |
Mar 23, 2010 |
8609152 |
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14088442 |
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12564878 |
Sep 22, 2009 |
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12730119 |
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11137248 |
May 24, 2005 |
7611738 |
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12564878 |
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Current U.S.
Class: |
514/27 |
Current CPC
Class: |
A61K 31/7048 20130101;
A23L 33/105 20160801; A61K 31/352 20130101; A61K 36/185 20130101;
A61K 2300/00 20130101; A61P 31/16 20180101; A61K 31/7048 20130101;
A61P 35/00 20180101; A61K 9/0095 20130101 |
Class at
Publication: |
514/27 |
International
Class: |
A61K 31/7048 20060101
A61K031/7048; A61K 31/352 20060101 A61K031/352 |
Claims
1. A method of treating a viral infection in a subject to alleviate
one or more symptoms, the method comprising: selecting a subject
having a viral infection, and administering to said subject an
effective amount of a polyphenol composition extracted from one or
more pomegranate solids, wherein said polyphenol composition
comprises punicalagin and punicalin, wherein a ratio of punicalagin
and punicalin to ellagic acid and ellagic acid glycosides in said
polyphenol composition by weight is greater than about 0.16.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of a U.S. patent
application Ser. No. 12/730,119 filed on Mar. 23, 2010 which is a
continuation-in-part of prior-filed and co-pending U.S. patent
application Ser. No. 12/564,878 entitled PROCESS FOR EXTRACTING
PHYTOCHEMICALS FROM POMEGRANATE SOLIDS AND COMPOSITIONS AND METHODS
OF USE THEREOF, filed Sep. 22, 2009, which is a continuation of
U.S. patent application Ser. No. 11/137,248, filed May 24, 2005,
(now U.S. Pat. No. 7,611,738), and this application is also a
continuation-in-part of prior-filed and co-pending U.S. patent
application Ser. No. 11/757,320 entitled METHOD OF USING
COMPOSITION COMPRISING POMEGRANATE EXTRACTS AGAINST INFLUENZA,
filed Jun. 1, 2007, which claims benefit to U.S. Provisional patent
application Ser. No. 60/809,859, filed Jun. 1, 2006, all of the
above which are hereby incorporated by reference in their
entirety.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates generally to pomegranate
extracts, and more particularly, to compositions and methods for
extracting and using phytochemicals for the treatment of
influenza.
[0004] 2. Description of the Related Art
[0005] It is well-known that fruits and vegetables are an essential
part of a healthy diet. Chief, among the reasons, is that fruits
and vegetables are rich sources of important phytochemicals, which
provide essential nutrients and enhance the body's ability to
prevent and fight disease. There is a multitude of phytochemicals,
in unique combinations, in different fruits and vegetables, and
each functions differently in the body: as anti-oxidants, as
anti-allergenic, as anti-carcinogenic, as anti-inflammatory, as
anti-viral, and/or anti-proliferative.
[0006] The pomegranate has recently been acclaimed for its health
benefits and for its disease-fighting antioxidant potential.
Antioxidants are important because they are believed to protect the
body against free radicals, the harmful molecules that can cause
heart disease, premature aging, Alzheimer's disease, blindness, and
a variety of cancers.
[0007] Studies have shown that pomegranate juice has more
polyphenol antioxidants than any other drink, such as red wine,
green tea, blueberry juice, cranberry juice and orange juice.
Currently, the two common ways of consuming pomegranates are by
eating the fleshy arils of the pomegranate and by drinking the
juice obtained from the arils.
[0008] There are many kinds of antioxidants, some produced by the
body and others derived from the foods we eat. When the body's
natural antioxidant defenses are lowered, or greater amounts of
free radicals are being produced, the body becomes more dependent
upon food sources of antioxidants.
[0009] The importance of influenza viruses as worldwide pathogens
for humans and domestic animals is well recognized. Influenza is a
major cause of morbidity and death. According to the Office of
Technology Assessment of the US Congress, each year, in the United
States alone, "the flu" accounts for 110,000 hospitalizations, 1 to
3 billion dollars in direct costs, and 10 to 15 billion dollars in
indirect costs. Influenza has been established as a serious human
affliction that can cause localized epidemics and global pandemic
of acute respiratory infection. There is a growing concern for
potential pandemic outbreak of influenza virus from the strain
currently in birds in Asia or another influenza virus.
[0010] Avian influenza is caused by type A strains of influenza
virus. Avian influenza occurs throughout the world. Infected birds
may display a wide range of symptoms, from a mild illness to a
highly contagious fatal disease. The highly contagious disease is
caused by an especially virulent strain of influenza virus.
Infection by this strain is associated with a sudden onset of
severe symptoms, such as a lack of energy, decreased egg
production, soft shelled eggs, a swelling of the head, eyelids,
etc., nasal discharge, coughing or diarrhea, resulting in death. At
present, 15 subtypes have been identified that can infect birds but
only H7, H5 and H9 subtypes are associated with outbreaks. The
current Asian and British Columbia outbreaks are caused by a H5N1
and H7N3 strains, respectively. As discussed above, influenza
viruses are a public health concern because these viruses lack a
mechanism for proofreading nucleic acid replication as well as a
repair system for correcting such errors. Thus, influenza viruses
are especially prone to a high mutation rate during transcription.
Additionally, influenza viruses are able to exchange or swap
genetic material from other subtypes from different species, thus
allowing subtypes to cross the species barrier that normally
prevents the cross infection of species specific viruses from one
species to another unrelated species. This species barrier normally
prevents avian influenza virus strains from infecting humans, but
occasionally new strains may have genetic material from both avian
and human influenza virus strains. This exchange of genetic
material occurs when there is a close proximity between humans and
domestic poultry and swine. Swine may act as a reservoir for both
human and avian strains. Thus swine act as a natural incubator for
the emergence of new strains that can infect humans as well as
avian species.
[0011] As mentioned above, influenza is prone to minor changes
though genetic material to one or more of the major surface
antigens during replication. The so-called antigenic drift is
responsible for the seasonal epidemics because it can enable the
virus to infect persons with only partial immunity from a prior
exposure to the virus. Influenza A viruses are especially prone to
antigenic drift. Major changes in the H and N antigens result in
antigenic shift. Antigenic shift results in a new viral subtype and
it can cause major epidemics and pandemics due to minimal immunity
in population. Pandemics happen when a novel influenza virus
emerges that infects and can be efficiently transmitted between
humans. Hence, there is a need for a composition that provides for
one or more anti-viral mechanisms that are not affected by
antigenic drift.
[0012] Influenza viral infection may be associated with redox
changes characteristic of oxidative stress. A more oxidized
environment may favor further viral infection and stimulates viral
protein synthesis. Various studies suggest that apoptosis of
mammalian cells may be caused by alteration of intracellular redox
condition induced by influenza virus infection. It was recently
reported that certain antioxidants inhibited the growth of
influenza viruses in Madin-Darby canine kidney (MDCK) cells and in
another study, antioxidants inhibited the replication of viral
strains in peripheral blood lymphocytes. These substances have
proven useful in the field treating various illnesses; however
there has not been any progress in the creation of a prophylactic
method for use with antioxidant compounds. This lack of effect by
other antioxidants on viral titer might represent the difference in
biological availability of these compounds or that anti-viral
activity operates through mechanisms other than anti-oxidation.
Therefore, there exists a need in the field to provide a
prophylactic method for the reduction of the incidence of
contracting an illness caused by influenza virus.
BRIEF SUMMARY OF THE PREFERRED EMBODIMENTS
[0013] Methods are provided for producing an extract containing
phytochemicals from pomegranate solids. The pomegranate solids are
anyone or more of the group consisting of the pericarp, inner
membrane and seeds. The extract produced differs from
commercially-available pomegranate juice in that the extract is
substantially derived from the pomegranate solids, whereas
pomegranate juice is substantially derived from the sweet, fleshy
arils.
[0014] In one preferred embodiment, the method includes the
following steps. Anyone or a combination of the pericarp, inner
membrane and seeds are selected and a mixture is formed comprising
the pomegranate solids and an aqueous solution. The mixture is then
heated to about 60.degree. F. to 210.degree. F., preferably of
about 85.degree. F. to 185.degree. F. and optimally of about
110.degree. F. to 160.degree. F. Enzymes are added to the mixture
in an amount sufficient to at least partially degrade the
pomegranate solids and liberate phytochemicals from the plant
tissues and/or cells. Once liberated, the phytochemicals may react
and/or polymerize to create new phytochemical compounds or reaction
products. The residual insoluble solid materials are removed from
the mixture to provide an extract containing phytochemicals.
[0015] In another preferred embodiment, extracts containing
phytochemicals from a pomegranate are provided. Such extracts are
characterized by a significantly higher total polyphenol content,
particularly of the high molecular weight polyphenol (e.g.,
punicalagin), than is found in pomegranate juice. Such extracts may
be obtained from the methods disclosed herein.
[0016] In a further preferred embodiment, food products and
beverages are provided comprising the extract containing
phytochemicals from a pomegranate.
[0017] In yet a further preferred embodiment, compositions
comprising the extract containing phytochemicals from a pomegranate
are provided. Such compositions may be in form of tablets,
suspensions, implants, solutions, emulsions, capsules, powders,
syrups, liquid compositions, ointments, lotions, creams, pastes,
and gels. Such compositions may also be in form of pharmaceutical
preparations, nutritional supplements, vitamin supplements, food
additives, and food supplements.
[0018] In a further preferred embodiment, compositions containing
the extract and the pomegranate juice are provided. The combination
of the extract and pomegranate juice not only produces a
composition having a higher total polyphenol content, as compared
to the pomegranate juice alone, but it also provides the broad
spectrum of the different polyphenols which predominate the
pomegranate juice and extract.
[0019] It is further an object of certain embodiments of the
invention to provide a method for reducing the incidence of
contracting an illness caused by influenza viruses.
[0020] In the first aspect, the present invention relates to a
method for the prophylactic use of an anti-viral composition to
reduce the incidence of contracting an illness. The method
comprises the steps of administering to a subject that has been, or
will be, exposed to an illness caused by an influenza virus, an
amount of an anti-viral composition having a composition extracted
from fruits of pomegranate; and optionally an acceptable carrier.
The amount of anti-viral composition is effective, when
administered, to reduce the incidence of contracting the
illness.
[0021] In a second aspect of the invention, a prophylactic
anti-viral composition having ingredients obtainable from
pomegranate fruit is disclosed. The anti-viral composition is
effective, when administered as a nasal spray or as an inhalant
spray to a subject that has been, or will be, exposed to an illness
caused by influenza virus, to reduce the incidence of contracting
said illness.
[0022] These and various other advantages and features of novelty
that characterize the invention are pointed out with particularity
in the claims annexed hereto and forming a part hereof. However,
for a better understanding of the invention, its advantages, and
the objects obtained by its use, reference should be made to the
accompanying descriptive matter, in which there is described a
preferred embodiment of the invention.
[0023] Other objects, features and advantages of the present
invention will become apparent to those skilled in the art from the
following detailed description. It is to be understood, however,
that the detailed description and specific examples, while
indicating preferred embodiments of the present invention, are
given by way of illustration and not limitation. Many changes and
modifications within the scope of the present invention may be made
without departing from the spirit thereof, and the invention
includes all such modifications and equivalents thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] The above and other aspects, features and advantages of the
invention will be more apparent from the following more partidcular
description thereof, presented in conjunction with the following
drawings wherein:
[0025] FIG. 1 is illustrates inhibition of viral replication by PJ
after and during viral absorption into cells.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0026] As used herein, the term "phytochemicals" refers
collectively to compounds which are naturally-occurring in the
pomegranate and to reaction products and metabolites of these
compounds, which are considered to have a beneficial effect on the
human health. Examples of such phytochemicals include, but are not
limited to polyphenols, estrogens and phytoestrogens.
[0027] As used herein, the term "polyphenols" refers generally to a
family of naturally-occurring compounds in the pomegranate and
includes phenols and polyphenols. Phenols are a class of chemical
compounds consisting of a single phenol unit in their structure.
Although similar to alcohols, phenols have unique properties
including relatively higher acidities due to the aromatic ring
tightly coupled to the oxygen and a relatively loose bond between
the oxygen and the hydrogen. Examples of phenolic compounds within
this group include ellagic acid and gallic acid. Polyphenols are a
group of compounds, characterized by the presence of more than one
phenolic group. Polyphenols include tannins (e.g., ellagitannins
and gallotannins), flavonoids (e.g., anthocyanins and isoflavones)
and stilbenes (e.g., resveratrol).
[0028] As used herein, the term "pomegranate juice" refers to the
juice that is substantially obtained from the arils of the
pomegranate.
[0029] As used herein, the term "pomegranate solids" refers to
anyone or a combination of the pericarp, the inner membrane and
seeds of a pomegranate.
[0030] It has been surprisingly discovered that extracts obtained
from the pomegranate solids, in accordance with the methods
disclosed herein, have a substantially higher total polyphenol
content than is found in the juice from the pomegranate arils. This
is particularly true with respect to the higher molecular weight
polyphenols and, in particular, punicalagin.
[0031] Punicalagin is a powerful antioxidant, protecting
cardiovascular function and accurate cellular replication. Thus,
punicalagin is responsible, in part, for the high antioxidant
activity of the extract. While the antioxidant and other beneficial
health effects of the extract are due to the presence of
polyphenols, the presence of other phytochemical compounds in the
extract, or the synergistic effect of these phytochemicals, may
also be responsible for the anti-oxidant and other beneficial
health effects of the extract.
[0032] In addition to punicalagin, other high molecular weight
polyphenols have been characterized in the extract of pomegranate
solids. These high molecular weight polyphenols include
ellagitannin and other hydrolysable tannins, such as punicacortein
A, punicalin, pedunculagin, and gallotanin dimmers and trimers.
[0033] Moreover, a large number of anthocyanins have been
characterized in the extract of the pomegranate solids. Examples of
the anthocyanins include pelargonidin 3-glucoside, cyaniding
3-glucoside, delphinidin 3-glucoside, pelargonidin 3,5-diglucoside,
cyaniding 3,5-diglucoside, and delphinidin 3,5-diglucoside.
Although these anthocyanins have been characterized in both the
pomegranate juice and the extract, these lower molecular weight
polyphenols comprise a higher proportion of the total polyphenol
content in pomegranate juice (approximately 50%) than in the
extract.
[0034] Accordingly, methods are provided for producing an extract
containing phytochemicals from pomegranate solids. The extract
produced from the methods disclosed herein differ from the
commercially-available pomegranate juice in that the extract is
substantially derived from the pomegranate solids, whereas
pomegranate juice is substantially derived from the sweet, fleshy
arils that surround the pomegranate seed. The extract is
characterized as containing polyphenols and, particularly, high
molecular weight polyphenols, such as punicalagin.
[0035] In one preferred embodiment, the method comprises providing
anyone or a combination of pomegranate solids selected from the
group consisting of the pericarp, inner membrane and seeds and
creating a mixture comprising the pomegranate solids in an aqueous
solution. In a preferred embodiment, the mixture of the pomegranate
solids is created by adding water in an amount that is about 20-80%
w/v, and more preferably about 50% w/v, of the pomegranate solids.
The mixture is preferably crushed or milled to create a rough grind
of pomegranate solids dispersed in the aqueous solution.
[0036] The mixture is then heated to a temperature of about
60.degree. F. to 210.degree. F., preferably of about 85.degree. F.
to 185.degree. F. and optimally of about 110.degree. F. to
160.degree. F. The temperature to which the mixture is heated
depends upon the selection of enzymes, or combination of enzymes,
that is added to the mixture. Preferably, the mixture is heated to
a temperature that permits the maximum catalysis of the enzyme or
combination of enzymes.
[0037] Alternatively, enzymes may be added before the mixture is
heated. Thus, the order of the steps of heating the mixture and
adding the enzymes is not critical, so long as the mixture is
heated to a temperature that permits the enzymes to at least
partially degrade the pomegranate solids and liberate
phytochemicals from the plant tissues and/or cells. Once liberated,
the phytochemicals may react and/or polymerize to create new
phytochemical compounds or reaction products.
[0038] Enzymes suitable for use in accordance with this embodiment
include those which are capable of at least partially degrading the
plant tissue or cells to liberate the phytochemicals from the
pomegranate solids. Such enzymes include anyone or a combination of
pectinase, cellulase, hemicellulase, amylase, arabanase, and other
hydrolyzing enzymes, to name a few. The enzymes added to the
mixture may be naturally-occurring or synthetic. They may be
derived from any one or a combination of sources, such as animal,
plant, fungal, and bacterial sources. The amount of the enzyme or
combination of enzymes added to the mixture depends on the
temperature of the mixture and the amount of pomegranate solids
present in the mixture.
[0039] After enzymes are added, the mixture is maintained at a
temperature for a time sufficient to allow at least partial
degradation of the pomegranate solids. The temperature and length
of time required depends on the type of enzymes added to the
mixture, the rate of enzyme catalysis and the amount of the
pomegranate solids contained in the mixture.
[0040] Thus, in one preferred embodiment, a combination of
pectinase, cellulase and hemicellulase enzymes are added to the
mixture, which is heated to a temperature of about 60.degree. F. to
210.degree. F., preferably about 110.degree. F. to 160.degree. F.,
and optimally of about 120.degree. F. The mixture is maintained at
these temperatures, preferably with agitation or stirring, for
about 45-195 minutes, preferably for about 45-75 minutes, and
optimally for about 60 minutes.
[0041] After the enzymes have at least partially degraded the
pomegranate solids, the residual insoluble solid materials are
removed from the mixture. Optionally, a clarification agent, such
as bentonite, may be added before the step of removing the residual
insoluble materials from the mixture. The removal of residual
insoluble materials from the mixture may be accomplished by
filtration, centrifugation, chromatographic techniques, and other
techniques. Filtration techniques suitable for the practice of the
methods disclosed herein include micro-filtration at a molecular
weight cut-off of at least 1,000 Da, preferably of about 4,500 Da,
and optimally of about 5,500 Da.
[0042] The resulting liquid extract may be concentrated in an
evaporator under vacuum to about 50-90 Brix (Bx), preferably to
about 60-80 Bx, and optimally to about 70 Bx, and pasteurized at a
temperature and for a length of time sufficient to kill
microorganisms that could cause disease, spoilage or undesired
fermentation. In one preferred embodiment, the extract may be
pasteurized at a temperature of about 140.degree. F.-280.degree.
F., preferably of about 195.degree. F.-240.degree. F., and
optimally of about 205.degree. F. The pasteurization may also
denature the remaining enzymes that were added to the mixture.
[0043] In another preferred embodiment, extracts containing
phytochemicals from a pomegranate are provided. Such extracts are
characterized by a significantly higher total polyphenol content,
particularly of the high molecular weight polyphenol (e.g.,
punicalagin), than is found in pomegranate juice. Such extracts may
be obtained from the methods disclosed herein. In a further
preferred embodiment, extracts containing phytochemicals,
polyphenols, punicalagin, punicalin, ellagic acid, and metabolite
thereof are provided.
[0044] In yet another preferred embodiment, food products and
beverages are provided comprising the extract containing
phytochemicals from a pomegranate. For example, due to the
significantly higher total polyphenol content in the extract, an 8
oz sports beverage containing 0.33 oz of the extract may be
formulated to deliver the same total polyphenols as a 20 oz
single-strength pomegranate juice. The polyphenol content of
pomegranate juice is approximately about 1 to 2.25 mg/mL and the
amount of polyphenols present in 20 oz of juice is approximately
567 to 1,256 mg. In contrast, the extract may contain a polyphenol
content of about 60 to 120 mg/mL, depending on the method
employed.` Thus only 0.33 oz of the 70 Bx extract would be needed
to provide the equivalent amount of polyphenols in 20 oz of the
juice.
[0045] In a further preferred embodiment, compositions comprising
the extract containing phytochemicals from a pomegranate are
provided. The compositions may be formulated in the form of
tablets, suspensions, implants, solutions, emulsions, capsules,
powders, syrups, liquid compositions, ointments, lotions, creams,
pastes, gels, and the like.
[0046] The compositions may also be prepared in forms suitable for
use as pharmaceutical preparations, nutritional supplements,
vitamin supplements, food supplements, and food additives. As such,
the compositions may optionally include a suitable carrier or
excipient.
[0047] Suitable carriers or excipients are inert ingredients and
include, by way of example, fillers, e.g. sugars such as lactose,
glucose or sucrose, sugar alcohols such as mannitol, sorbitol or
xylitol, starch such as wheat, corn or potato starch, modified
starch or sodium starch glycolate, lubricants such as talc,
magnesium stearate, calcium stearate, colloidal silica or stearic
acid, and binders such as polyvinylpyrrolidone, cellulose
derivatives, carboxymethyl cellulose, hydroxylpropyl cellulose,
hydroxypropylmethyl cellulose, methyl cellulose or gelatin.
Conventional procedures for preparing such compositions in
appropriate,dosage forms of the extract may be utilized. Such
compositions may be administered orally or parenterally employing
liquid form preparations containing the extract.
[0048] The compositions may be administered orally, in appropriate
dosage units of the extract in a pharmaceutically acceptable
carrier or excipient. Thus, the compositions may be formulated into
solid or liquid preparations, such as capsules, pills, tablets,
powders, solutions, suspension, or emulsions and may be prepared
according to methods known in the art for the manufacture of such
compositions. The solid unit dosage forms may be in form of a hard
or soft shelled gelatin capsule containing the extract and a
suitable carrier or excipient.
[0049] The composition may also be administered parenterally as
injectable dosages in a physiologically acceptable carrier.
Parenteral administration may be subcutaneous, intravenous,
intramuscular, or interperitoneally.
[0050] The effective amount of a composition is the amount or
dosage unit of the extract sufficient to achieve the intended
beneficial health results. Accordingly, the effective amount of the
composition to be administered depends on considerations such as
the dosage unit employed, the mode of administration, the period of
treatment, the age, sex and weight of the person treated and the
nature and extent of the condition treated. The effective amount
can readily be determined based upon standard techniques known to
evaluate whether the intended effect of the composition has been
achieved, by standard toxicity tests and by standard
pharmacological assays.
[0051] In a further preferred embodiment, compositions containing
the extract and the pomegranate juice are provided. The combination
of the extract and pomegranate juice not only produces a
composition having a higher total polyphenol content, as compared
to the pomegranate juice alone, but it also provides the broad
spectrum of the different polyphenols which predominate the
pomegranate juice and extract, for example the lower molecular
weight polyphenols (e.g., anthocyanins) which is present in greater
quantities in the pomegranate juice and the higher molecular weight
polyphenols (e.g., punicalagin, punicalin, ellagic acid glycosides,
ellagic acid polyphenols, ellagitannin and other hydrolysable
tannins, such as punicacortein A, punicalin, pedunculagin, and
gallotanin dimmers and trimers).
[0052] In yet a further preferred embodiment, methods are provided
for ameliorating disease conditions in a subject by administering
to the subject an effective amount of the composition suitable for
use as a pharmaceutical or nutritional preparation.
[0053] Thus, in one preferred embodiment, methods are provided for
formulating a composition suitable for use as a pharmaceutical or
nutritional preparation for improving the health of a subject
comprising obtaining an extract containing phytochemicals from a
pomegranate and admixing an effective amount of the extract with a
suitable carrier or excipient. In another preferred embodiment,
methods are provided for treating a polyphenol-mediated condition
in a subject comprising selecting a subject having a
polyphenol-mediated condition and administering to the subject an
effective amount of the composition comprising the extract.
[0054] The composition of the present invention may be used to
treat influenza viral infection, since the composition of the
present invention has significant antiviral properties as
demonstrated by the examples of this application. The composition
of the present invention may also be used as a therapeutic
composition to treat one or more symptoms of a viral infection,
including, but not limited to, sore throat, congestion, laryngitis,
mucositis, and/or mucous membrane inflammation by administration to
a subject suffering from one or more of these symptoms or
ailments.
[0055] The composition of the present invention may also be
employed to reduce the incidence of contracting an illness. In this
application of the composition of the present invention, a safe and
effective amount of the composition of the present invention is
administered to a mammal or a bird that has been or will be exposed
to an illness caused by a microbe, to reduce the incidence of
contracting said illness, relative to a mammal or a bird that has
been or will be exposed to an illness caused by a microbe to which
the composition of the present invention has not been
administered.
[0056] The composition of the present invention may also be
formulated with an acceptable carrier. The acceptable carrier may
include, but is not limited to: (a) carbohydrates including
sweeteners, fructose, sucrose, sugar, dextrose, starch, lactose,
maltose, maltodextrins, corn syrup solids, honey solids, commercial
tablet nutritional supplements (b) sugar alcohols including
mannitol, sorbitol and xylitol; and (c) various relatively
insoluble excipients including dicalcium phosphate, calcium
sulfate, calcium carbonate, microcrystalline cellulose and other
tableting ingredients.
[0057] The composition of the present invention may also be
formulated into a nasal aerosol or inhalant composition. Such a
composition may be prepared using well-known techniques. For these
types of formulations, suitable carriers may include the following
ingredients: saline with one or more preservatives, absorption
promoters to enhance bioavailability, fluorocarbons, and/or
conventional solubilizing or dispersion agents.
[0058] For the purpose of the invention, an extract from
pomegranate may be an extract from the whole pomegranate fruit or
from any constituents of pomegranate fruit. Examples of
constituents of pomegranate fruit that may be used to make the
extract of the invention include, but are not limited to, juice,
seed, and the inner and outer peel of pomegranate fruit. In one
embodiment of the invention, the extract is the juice extract of
whole pomegranate fruit. In another embodiment of the invention,
the extract is from the inner or outer peel of pomegranate fruit.
In a further embodiment of the invention, the extract may be a
mixture of two or more extracts of the whole pomegranate or any
constituents of pomegranate.
[0059] In general, any methods that may produce pomegranate juice
that naturally occurs in pomegranate may be used. For the purpose
of the invention, the juice may be concentrated or diluted from its
natural concentration. The juice may also be mixed with extracts of
other constituents of pomegranate to vary the composition. Methods
of making the extract, including the juice from whole pomegranate
fruits are described in U.S. Pat. No. 6,977,089 entitled "METHODS
OF USING POMEGRANATE EXTRACTS FOR CAUSING REGRESSION IN LESIONS DUE
TO ARTERIOSCLEROSIS" which is incorporated herein by reference in
its entirety.
[0060] Extracts from the constituents of pomegranate, i.e., seeds
or the inner or outer peel, may be made by various methods. For
example, the seeds or the inner or outer peel of pomegranate may be
diluted in water and the extract may be made by crushing,
squeezing, or extensive vortexing. The insoluble materials of the
extract may be separated from the soluble supernatant of the
extract. In some instances, the supernatant of the extract is used
for the purpose of the invention, although any oily, lipidic
fraction of the extract may also be used. The extract from
constituents of pomegranate may be concentrated or diluted, or
mixed with each other or with pomegranate juice extract.
[0061] In accordance with one embodiment of the invention, the
extract of the present invention may be prepared by a process
including the steps of: (a) crushing and squeezing the whole fruits
of the pomegranate, including the inner and outer peels and the
seeds, to yield a juice component and an insoluble by-product
component, and (b) separating the juice component from the
insoluble by-product component. The juice component may be used as
a juice extract of the invention. The insoluble by-product
component may be resuspended in an aqueous medium, such as, but not
limited to, water or alcohol, and be further crushed, squeezed, and
mixed to yield a soluble portion and an insoluble portion. Then the
soluble portion may be separated from the insoluble portion to
produce the extract of the constituents of the invention.
Alternatively, the soluble portion may be combined with the juice
extract to produce the extract of the invention.
[0062] In one embodiment of the invention, the whole fruit of the
pomegranate may be enzymatically treated to improve extraction and
filtration. For example, pectinase may be used to treat the whole
fruit to prevent the formation of pectin gels. Other enzymes may
also be used as long as they can improve extraction and filtration
of the extract of the invention.
[0063] The extract of pomegranate used in accordance with one or
more embodiments of the invention may be in a liquid or solid form.
In accordance with one embodiment of the invention, a solid form of
the extract may be made by lyophilizing the liquid extract of the
invention. Alternatively, the constituents of the pomegranate, such
as seeds, inner or outer peels, or any insoluble portion discussed
above, may be processed directly to form the solid form of the
extract of the invention. For example, the constituents of the
pomegranate may be dried, and processed into powder or pill forms
to be used directly as the solid form of the extract of the
invention.
[0064] Compositions of one or more embodiments of the invention may
be a variety of kinds, including, but not limited to, nutritional
supplements, pharmaceutical preparations, vitamin supplements, food
additives, or foods supplements. Compositions of the invention may
be in convenient dosage forms, including, but not limited to,
tablets, suspensions, implants, solutions, emulsions, capsules,
powders, syrups, liquid compositions, ointments, lotions, creams,
pastes, gels, or the like.
[0065] Compositions of one or more embodiments of the invention may
include a carrier. Depending on the kind of compositions of the
invention, a carrier may be a dietary suitable carrier or a
pharmaceutically acceptable carrier, as long as it is compatible
with the particular kind of compositions of the invention. Examples
of a dietary suitable carrier include, but are not limited to,
dietary suitable excipients, diluents, and carriers. Examples of a
pharmaceutically acceptable carrier include, but are not limited
to, biocompatible vehicles, adjuvants, additives, and diluents to
achieve a composition usable as a dosage form. As used herein, the
terms "pharmaceutically acceptable," "physiologically tolerable,"
and grammatical variations thereof, as they refer to compositions,
carriers, diluents, and reagents, are used interchangeably and
represent that the materials are capable of administration to or
upon a mammal without the production of undesirable physiological
effects.
[0066] The compositions of one or more embodiments of the invention
may be used alone or in combination with other biologically active
ingredients. A composition of embodiments of the invention, alone
or in combination with other active ingredients may be administered
to a subject in a single dose or multiple doses over a period of
time, generally by oral administration. Various administration
patterns will be apparent to those skilled in the art. The dosage
ranges for the administration of the compositions of the invention
are those large enough to produce the desired effect. The dosage
should not be so large as to cause any adverse side effects, such
as unwanted cross-reactions and the like. Generally, the dosage
will vary with the age, weight, sex, condition, and extent of a
condition in a subject, and the intended purpose. The dosage can be
determined by one of skill in the art without undue
experimentation. The dosage can be adjusted in the event of any
counter indications, tolerance, or similar conditions. Those of
skill in the art can readily evaluate such factors and, based on
this information, determine the particular effective concentration
of a composition of the invention to be used for an intended
purpose.
[0067] In one embodiment of the invention, a composition contains
the extract of pomegranate in a dosage unit in an amount that
contains at least 30 to 10,000 parts per million or 30 to 3,000 mg
of polyphenols. For the purpose of the invention, polyphenols are
those naturally present in the extract of pomegranate. It should be
appreciated that polyphenols are used herein as a measurement
marker for the amount of extract used in each dosage unit.
Polyphenols are not used herein as being indicative of the only
active, ingredients of the extract. It is possible, for example,
that other elements of the composition or the synergy of
polyphenols and other components of an extract of the invention,
are responsible for the activities of the extract.
[0068] The term "dosage unit" as used herein refers to physically
discrete units suitable as unitary dosages for subjects, each unit
containing a predetermined quantity of active material calculated
to produce the desired therapeutic effect in association with the
required diluent, e.g., a carrier or vehicle. The specifications
for the unit dose of this invention are dictated by and are
directly dependent on (a) the unique characteristics of the active
material and (b) the limitations inherent in the art of compounding
such active material for therapeutical use in subjects.
[0069] Prophylactic treatment is aimed at a subject that will soon
be exposed to a virus or has recently been exposed to a virus. Such
prophylactic treatment may be effective either alone, or to augment
a vaccine or another anti-viral drug. Prophylactic treatment may
also be used against viruses for which there are not yet a vaccine
available. In the case of prophylactic treatment, the composition
of the invention is administered to a subject that will be exposed
to a virus or has recently been exposed to a virus for the purpose
of reducing the incidence of active infection by the virus in that
subject.
[0070] In another aspect, the present invention relates to a method
of reducing, treating or preventing of at least one symptom or
adverse effect of viral infection by administering, to a subject
infected with a virus, a composition of the present invention,
including ingredients that can be obtained from pomegranate.
[0071] In the method, the subject may be a human, an in vitro cell
system, or an animal. Preferably, the subject is a mammal, more
preferably, a human. In the method, the virus that may be inhibited
by administration of the composition of the present invention
includes, among other viruses, rhinoviruses, influenza viruses,
West Nile virus, herpes simplex virus, HIV-1, HIV-2, adenovirus,
cornavirus, influenza virus, rubella virus, yellow fever virus and
respiratory syncytial virus (RSV). In a preferred embodiment, the
viruses that may be inhibited by administration of the composition
include at least Influenza A/Hong Kong/8/68 (H3N2).
[0072] Alternatively, the subject may be a member of the bird
(Avian) species, which includes the common commercial poultry
birds: turkeys, ducks, geese and chickens, less commonly the
ostrich as well as other bird species that are commonly kept as
house pets, for example canaries and parrots. The composition may
be administered by directly spraying the composition into the nasal
passage of the bird or the composition may be administered by
creating a mist through which the birds walk. Thus, the composition
may be given prophylactically to act in a virucidal or virustatic
manner. Alternatively, the composition may be used to reduce the
transmissivity of the virus.
[0073] The symptoms, caused by a viral infection, that may be
treated, reduced, or at least partially prevented by this method of
the present invention, may include one or more of headache, joint
pain, fever, cough, sneezing, muscle ache, running nose, dry mouth,
dizziness, and other symptoms related to viral infection. In birds,
these symptoms include a lack of energy, decreased egg production,
soft shelled eggs, a swelling of the head, eyelids, etc., nasal
discharge, coughing or diarrhea.
[0074] The effective amount of the composition will vary depending
on such factors as the subject being treated, the particular mode
of administration, the activity of the particular active
ingredients employed, the age, bodyweight, general health, sex and
diet of the subject, time of administration, rate of excretion, the
particular combination of ingredients employed, the total content
of the main ingredient of the composition, and the severity of the
illness or symptom. It is within the skill of the person of
ordinary skill in the art to account for these factors.
[0075] When the composition is administered as a spray, the amounts
each of the active ingredients may be reduced as the spray
composition delivers the active ingredients more directly to the
location where they are needed, as compared to a lozenge or capsule
for example.
[0076] The following examples are intended to illustrate, but not
to limit, the scope of the invention. Indeed, those of ordinary
skill in the art can readily envision and produce further
embodiments, based on the teachings herein, without undue
experimentation. The scope of invention is to be determined by the
claims appended hereto.
EXAMPLE 1
[0077] Production of Liquid Extract from Pomegranate Solids
[0078] The starting material for the production of the extract is
the pomegranate solids, which generally comprise the pericarp, the
inner membrane and seeds of the pomegranate. The pomegranate solids
were obtained and collected after the primary juice from the arils
had been substantially expelled or otherwise removed from the
pomegranate by pressing, crushing, or other methods known to the
art for extracting pomegranate juice.
[0079] The pomegranate solids were then transferred to three Reitz
Mills with 3/8-inch screens. The material was milled to a fine
puree and heated to approximately 125.degree. F. This step, coupled
with the following enzyme addition, assisted in breaking down the
colloidal structure of the remaining pomegranate solids, thereby
releasing the remaining soluble solids.
[0080] The mixture was heated to a temperature of about 125.degree.
F. for two hours. Three enzymes were added to the mixture:
pectinase (Rohapect.RTM. DA6L), cellulase/pectinase (Rohapect.RTM.
CL), and hemi-cellulase/pectinase (Rohapect.RTM. B1L). These
enzymes were used to liberate the remaining pomegranate soluble
solids, such as sugars, minerals, anthocyanins, and remaining
polyphenols.
[0081] The mixture was then pumped from the extraction plant to the
primary processing plant where it was held in the mash treatment
tanks for approximately one hour. After one hour, 50-100 pounds of
bentonite in a 125 galloon water slurry, per 8,000 gallons of the
mixture, was added for protein removal. The treated mixture was
then passed through a Westphalia 755 Decanter for removal of
solids. The residual insoluble material was discharged as
waste.
[0082] The remaining liquid extract was processed in a Schmidt
evaporator. In this step, the extract was stripped and rectified.
In addition, the liquid extract was pre-concentrated and then
pasteurized to 205.degree. F. for 45 seconds. The liquid extract
then exited the evaporator and was filtered on Koch
Micro-Filtration membranes at a 4,500 Da molecular weight cut-off
for liquid extract soluble solids.
[0083] The liquid extract then re-entered the evaporator for final
concentration. Initial heat on this step was about 185-195.degree.
F. At about 70 Bx, the liquid extract was cooled to less than about
45.degree. F. and pumped to the concentrate batching room where it
was blended and standardized.
EXAMPLE 2
Comparison of Polyphenol Content in Extracts of Pomegranate Solids
and in Pomegranate Juice
[0084] The concentrations of punicalagin, punicalin, ellagic acid
glycosides, and ellagic acid polyphenols in the pomegranate juice
and the pomegranate extract were analyzed and compared in a
University study.
[0085] All samples (50 mL injection volume) were filtered (0.22 mm)
and analyzed on a Novapak (Waters Corp.) C-18 column, 150.times.3.9
mm i.d., 5 mm. The mobile phase, solvent A (2% CH3COOH/H2O) and
solvent B (2% aqueous CH3COOH/CH3OH) was used under linear gradient
conditions starting with 99% solvent A in solvent B to 40% solvent
A in solvent B over 40 minutes, hold time, 5 minutes with a flow
rate of 1 mL/min. All compounds were detected at 254 nm, and at 378
nm (punicalagins) and 366 (ellagic acid) for quantification.
[00060] Table 1 shows a side-by-side comparison of the
concentration of the polyphenols punicalagins, punicalin, ellagic
acid glycosides, and ellagic acid in the pomegranate extract and
the pomegranate juice.
TABLE-US-00001 TABLE 1 Pomegranate Extract Pomegranate Juice
Concentration Concentration Compound Name (mg/ml) (mg/ml)
Punicalagin (.beta. isomer) 4.79 0.02 Punicalagin (.alpha. isomer)
21.80 0.15 Punicalin 3.62 NA Ellagic Acid 19.65 0.33 Glycosides
Ellagic Acid 18 0.74 Total 67.86 1.24
[0086] The pomegranate extract concentration of table 1 is a
polyphenol composition for comprising punicalagin and punicalin,
wherein a ratio of punicalagin and punicalin to ellagic acid and
ellagic acid glycosides by weight is greater than that found in the
pomegranate juice contentration, which is about 0.16. As shown in
Table 1, the ratio of punicalagin and punicalin to ellagic acid and
ellagic acid glycosides in the pomegranate extract concentration,
by weight, is at least about 3:2. Furthermore, the pomegranate
extract contentration of Table 1 has a ratio of punicalagin and
punicalin to free ellagic acid of at least 4:1.
[0087] Although other polyphenols are present in both the
pomegranate extract and juice, and this example highlights the
unexpected and surprising results in that significantly higher
concentrations of polyphenols, particularly of punicalagin, are
present in the pomegranate extract than in the pomegranate juice.
Table 1 shows a total punicalagin (for both a and .beta. isomers)
concentration for the pomegranate extract that is over 26-fold
greater than for the pomegranate juice.
EXAMPLE 3
Dosages
[0088] While both pomegranate juice and pomegranate solid extract
contain various types of the anti-oxidant polyphenols, pomegranate
solid extract contains a higher total polyphenol content than the
pomegranate juice.
[0089] In preferred one embodiment, the composition may be in form
of a liquid comprising the extract and pomegranate juice. The total
polyphenol content provided by the liquid may be varied by the
changing the amount of the pomegranate extract and pomegranate
juice contained in the liquid. Table 2 provides examples of the
formulations of the liquid composition and the total polyphenol
content in the formulations relative to the total polyphenol
content in standard pomegranate juice. For example, 8 oz of
pomegranate juice by mouth daily, Wonderful variety, is equivalent
to 1.5 mmol of total polyphenols per day.
TABLE-US-00002 TABLE 2 Extract/Pomegranate Juice Liquid Pomegranate
Pomegranate Composition Extract (oz) Juice (oz) Formulation 1 0.13
0.8 2x polyphenol content Formulation 2 0.26 0.8 3x polyphenol
content Formulation 3 0.39 0.8 4x polyphenol content Formulation 4
0.52 0.8 5x polyphenol content
[0090] For purposes of this embodiment, the effective amount of the
extract that is administered to the patient is at least 0.13 oz (or
an equivalent unit or measurement) of the extract administered
daily, whether the extract is provided alone or in a composition.
The dosage of the extract may be increased by administering a
greater dosage or increasing the frequency at which the extract is
administered.
[0091] In addition to the liquid compositions containing the
extract, the extract may also be administered in a solid form, such
as pharmaceutical or nutritional preparation that comprises the
extract and a pharmaceutically acceptable carrier or excipient.
EXAMPLE 4
A Composition of the Present Invention
[0092] Pomegranate juice (PJ) extract (Wonderful variety, POM
Wonderful, Los Angeles) was used in studies to confirm the benefits
of the treatment set forth herein for influenza. Pomegranates were
hand-picked, washed, chilled to 4.degree. C., and stored in tanks
The fruit was then crushed, squeezed, and treated enzymatically
with pectinase to yield the juice and byproducts, which included
the inner and outer peels and the seeds. Pectinase hydrolyzes
-1,4-galacturonide bonds in pectin, improving extraction and
filtration, and prevents formation of pectin gels. Flavonoids
constitute 40% (anthocyanins, catechins, and phenols) of total
polyphenols in PJ. More complex polyphenols are also present in the
juice. The PJ was filtered, pasteurized, concentrated, and stored
at -20.degree. C. until use.
EXAMPLE 5
Treatment of Sore Throat
[0093] Each of seven human subjects, suffering from sore throats,
ingested one cup of PJ prepared according to Example 1 every four
hours.
[0094] The human subjects that were treated reported complete
relief from the symptoms of their sore throats after ingesting from
1 to 6 cups of PJ. It was also found that each cup of PJ can
provide relief from a sore throat for up to 6 hours.
[0095] Transmission of cold/flu virus is from person to person
usually by personal contact by shaking hands and wipe nose or rub
eyes or getting sneeze on and the like.
[0096] Mechanism of infection: Upon entry through the nose and the
eyes, the virus enters the cells lining the nasopharynx (the area
of the upper throat that lies behind the nose) and multiples
rapidly. Once viruses reach a high concentration cold/flu symptoms
(related to inflammation) rapidly appear.
[0097] The nasopharynx is area typically for build high
concentration of viruses needed to cause cold or flu symptoms. This
build-up of viral concentration is because they are either: (1)
trapped by the mucus membranes, which is a natural defense
mechanism and/or (2) the viruses concentrate on the outer part of
the membranes, where they are less exposed to normal body
temperature (since they cannot survive elevated temperatures such
as body temperature). The cold/flu viruses are vulnerable on the
top of nasopharynx membranes on the upper throat.
[0098] Human subjects infected with cold/flu virus not showing
symptoms are selected in this study. It is desirable keep the virus
concentrations low in the upper throat/nasopharynx during the first
few days of infection. This prevents the body's defense mechanism,
e.g. sneezing, runny nose, fever, malaise and cough, from turning
on as the symptoms may confound the study.
[0099] The oral consumption of PJ can strongly inhibit the viruses
on the nasopharynx since it coats the lining of the
throat/nasopharynx with PJ. The varying amount of PJ used to
inhibit viral replication on both surface and inner layers of the
cells are shown in FIG. 1.
[0100] It should be noted that the concentration of polyphenols in
PJ is over 100 times more concentrated than in the diluted PJ as
indicated in the FIG. 1. This high concentrated source of
polyphenols in PJ can immediately inhibit viral replication,
protein transport systems, and residual polyphenols are expected to
remain for some period in on the cell surface or mucosal layer
surrounding the throat/nasopharynx cells. So PJ is used to treat
the nasopharynx which disrupt viral replication and control
cold/flu symptom, typically when caught in time before viral levels
get too high.
[0101] Conclusion: Daily or multi-times per day consumption of PJ
may help keep viral concentrations at levels below threshold for
symptoms. This study indicates prevention during the first 1-2 day
of exposure.
[0102] The invention described and claimed herein is not to be
limited in scope by the specific embodiments herein disclosed,
since these embodiments are intended as illustrations of several
aspects of the invention. Any equivalent embodiments are intended
to be within the scope of this invention. Indeed, various
modifications of the invention in addition to those shown and
described herein will become apparent to those skilled in the art
from the foregoing description. Such modifications are also
intended to fall within the scope of the appended claims.
* * * * *