Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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FUNCTIONAL FOOD COMPOSITIONS AND METHODS
FIELD AND BACKGROUND OF THE INVENTION
The present invention relates to compositions including bioactive components
carried within
solid lipid particles dispersed in a food based carrier and to methods of
producing such
compositions.
Nutraceutical, a term combining the words "nutrition" and "pharmaceutical", is
a food or
food product that provides health and medical benefits, including the
prevention and
treatment of diseases. Neutraceuticals include, among other categories,
functional foods and
medical foods [see Chen et al. Trends in Food Science & Technology 17 (2006)
272-283;
and Lemes and McClements Trends in Food Science & Technology 20 (2009)].
Functional foods are consumed close to their natural state and include
components or
ingredients that provide a specific medical or physiological benefit beyond
(or in addition to)
the nutritional effects of the food. Garlic, which includes sulfur compounds,
and fish, which
include omega-3 fatty acids, are examples of functional foods. Functional
foods can be
enriched or fortified to restore or supplement the nutrient content or
physiological effect of
food.
Medical foods are formulated to be consumed or administered internally under
the
supervision of a physician and are intended for dietary management of a
disease or condition
for which there are specific nutritional requirements.
Incorporation of bioactive components such as vitamins, probiotics, bioactive
peptides, and
antioxidants into food based carriers provide a simple way to develop novel
functional foods
that may have physiological benefits or reduce the risks of diseases. However,
functional and
medical foods are oftentimes limited in as far as their bioactive agents
loading capacity (i.e,
the concentration of the bioactive agent in the food base carrier), especially
in cases where
the active agent can not be readily solubilized or emulsified in the food base
carrier.
Bioactive food components are biomolecules that are present in foods such as
fruits and
vegetables and which exhibit the capacity to modulate one or more metabolic
processes, and
thus can be used in the treatment of disease and promotion of better health.
Bioactive food components can be ingested as part of the source food or they
can be ingested
in purified form. In many cases it is unclear whether such components are
ingested at
effective dosages required for health benefits due to their limited
bioavailability, however.
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Methods of enhancing the bioavailability of bioactive food components are well
known in the
art and include formulations which are designed to increase the
gastrointestinal (GI)
absorption of the bioactive food components and/or limit their breakdown in
the GI tract or
body (see, Davis, DDT, Volume 10, Number 4, Feb. 2005).
Although such formulations can be used to increase bioavailability of
bioactive food
components, food derived biomolecules are oftentimes difficult to ingest due
to offensive
odors or smells and do not provide the nutritional value associated with the
food from which
the bioactive food components are derived.
Thus, there remains a need for composition that includes a nutritional base
and bioactive
components devoid of the above limitations.
The present invention successfully addresses the shortcomings of the presently
known
configurations by providing food-based physiologically-functional compositions
that are
capable of oral delivery of physiologically effective amounts of a bioactive
agent.
Unless otherwise defined, all technical and scientific terms used herein have
the same
meaning as commonly understood by one of ordinary skill in the art to which
this invention
belongs. Although methods and materials similar or equivalent to those
described herein can
be used in the practice or testing of the present invention, suitable methods
and materials are
described below. In case of conflict, the patent specification, including
definitions, will
control. In addition, the materials, methods, and examples are illustrative
only and not
intended to be limiting.
SUMMARY OF THE INVENTION
It is one object of the present invention to provide a method of producing a
functional food,
comprising:
melting a lipid solvent in the presence of a lipophilic bioactive agent;
heating the melted lipid solvent and bioactive agent to a temperature of at
least 65 C;
adding a food base carrier to said melted lipid solvent and bioactive agent;
and,
emulsifying said melted lipid solvent, said bioactive agent, and said food
base carrier
until an emulsion is formed, said emulsion comprising lipid particles
comprising
said bioactive agent.
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It is another object of the present invention to provide the method as defined
above, wherein
said step of heating the melted lipid solvent and bioactive agent to a
temperature of at least 65
C comprises a step of heating the melted lipid solvent and bioactive agent to
a temperature
of between 70 C and 90 C.
It is another object of the present invention to provide the method as defined
above, wherein
said step of emulsifying is at least partially effected simultaneously with a
step of cooling.
It is another object of the present invention to provide the method as defined
above, wherein
said step of cooling further comprises a step of cooling to about 45 C.
It is another object of the present invention to provide the method as defined
above, wherein
said step of cooling further comprises a step of cooling to a temperature at
least 20 C below
the emulsification temperature.
It is another object of the present invention to provide the method as defined
above, further
comprising heating an emulsifier to a temperature of at least 90 C in the
presence of said
bioactive agent prior to said step of melting said lipid solvent in the
presence of said
bioactive agent.
It is another object of the present invention to provide the method as defined
above, wherein
said emulsifier is chosen from the group consisting of PEG esters and sucrose
esters.
It is another object of the present invention to provide the method as defined
above, wherein
said emulsifier is a PEG ester chosen from PEG6000 esters, PEG 100 stearate,
and PEG40
stearate.
It is another object of the present invention to provide the method as defined
above, further
comprising a step of heating at least part of said food base carrier prior to
said step of adding
said food base carrier to said melted lipid solvent and bioactive agent.
It is another object of the present invention to provide the method as defined
above, wherein
said step of heating at least part of said food base carrier comprises heating
a first part of said
food base carrier to a first predetermined temperature and heating a second
part of said food
base carrier to a second predetermined temperature lower than said first
predetermined
temperature.
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It is another object of the present invention to provide the method as defined
above, wherein
said first predetermined temperature is at least 60 C and said second
predetermined
temperature is not above 50 C.
It is another object of the present invention to provide the method as defined
above, wherein
said step of adding a food base carrier further comprises adding said first
part of said food
base carrier, and further comprising a step of adding said second part of said
food base carrier
during said step of emulsifying, said step of adding said second part taking
place only when
the temperature of the other components being emulsified is below 60 C.
It is another object of the present invention to provide the method as defined
above, further
comprising:
heating a portion of said food base carrier in the presence of potassium
sorbate until a
solution forms;
cooling said solution to room temperature; and,
adding said solution to said melted lipid phase and bioactive agent.
It is another object of the present invention to provide the method as defined
above, wherein
said food base carrier is a semisolid at room temperature.
It is another object of the present invention to provide the method as defined
above, wherein
said food base carrier is a liquid at room temperature.
It is another object of the present invention to provide the method as defined
above, wherein
said bioactive agent comprises a bioactive food component.
It is another object of the present invention to provide the method as defined
above, wherein
said step of melting a lipid solvent in the presence of a lipophilic bioactive
agent comprises
melting a lipid solvent in the presence of a bioactive agent is chosen from
the group
consisting of curcumin, diindolylmethane, quercetin, diadezin, silymarin,
genistein, essential
fatty acids, and phytosterols.
It is another object of the present invention to provide the method as defined
above, wherein
said food base carrier is chosen from the group consisting of saccharide
syrups and
polysaccharide syrups.
It is another object of the present invention to provide the method as defined
above, wherein
said syrup is chosen from the group consisting of honey, date syrup, and maple
syrup.
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It is another object of the present invention to provide the method as defined
above, further
comprising a step of allowing at least part of said lipid to solidify into
particles within said
food base carrier.
It is another object of the present invention to provide the method as defined
above, further
comprising a step of dispersing homogeneously said particles within said food
base carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of allowing at least part of said lipid to solidify into particles
within said food base
carrier further comprises allowing at least part of said lipid to solidify
into microspheres
within said food base carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of allowing at least part of said lipid to solidify into
microspheres within said food
base carrier further comprises a step of allowing at least part of said lipid
to solidify into
microspheres of diameter between 0.5 gm and 5 gm within said food base
carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of allowing said lipid to solidify into particles within said food
base carrier further
comprises a step of allowing said lipid to solidify into particles within said
food base carrier
chosen from the group consisting of solid particles and semisolid particles.
It is another object of the present invention to provide the method as defined
above, wherein
at least 70% of said bioactive agent is associated with said particles.
It is another object of the present invention to provide the method as defined
above, wherein
at least 80% of said bioactive agent is associated with said particles.
It is another object of the present invention to provide the method as defined
above, wherein
between 90% and 95% of said bioactive agent is associated with said particles.
It is another object of the present invention to provide the method as defined
above, further
comprising a step of obtaining at least one lipid solvent chosen from the
group consisting of
hydrogenated castor oil; stearin; palm oil; high-omega-3 sage oil; pomegranate
oil; avocado
oil; olive oil; and any combination thereof.
It is another object of the present invention to provide the method as defined
above, further
comprising a step of mixing a bioactive agent with said food base carrier.
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It is another object of the present invention to provide the method as defined
above, wherein
said step of mixing a bioactive agent with said food base carrier further
comprises a step of
mixing a bioactive agent chosen from the group consisting of vitamins and
minerals with said
food base carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of mixing a bioactive agent with said food base carrier further
comprises a step of
mixing a hydrophilic bioactive agent with said food base carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of mixing a hydrophilic bioactive agent with said food base carrier
further
comprises a step of mixing a hydrophilic bioactive agent chosen from the group
consisting of
cobalamin, folate, and ferrous gluconate to said food base carrier with said
food base carrier.
It is another object of the present invention to provide a functional food
comprising a
lipophilic bioactive agent, a lipid, and a food base carrier, wherein said
lipid is at least
partially dispersed as particles within said food base carrier and said at
least part of lipophilic
bioactive agent is contained within said particles.
It is another object of the present invention to provide the functional food
as defined above,
wherein said particles constitute between 5% and 40% (w/w) of the food.
It is another object of the present invention to provide the functional food
as defined above,
wherein said lipophilic bioactive agent is a bioactive food component.
It is another object of the present invention to provide the functional food
as defined above,
wherein said lipophilic bioactive agent comprises at least one component
chosen from the
group consisting of curcumin, diindolylmethane, quercetin, diadezin,
silymarin, genistein,
essential fatty acids, and phytosterols.
It is another object of the present invention to provide the functional food
as defined above,
wherein said lipid is chosen from the group consisting of hydrogenated castor
oil; steam;
palm oil; high-omega-3 sage oil; pomegranate oil; avocado oil; olive oil; and
any
combination thereof.
It is another object of the present invention to provide the functional food
as defined above,
wherein said food base carrier is a liquid at room temperature.
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It is another object of the present invention to provide the functional food
as defined above,
wherein said food base carrier is a semisolid at room temperature.
It is another object of the present invention to provide the functional food
as defined above,
wherein said food base carrier is chosen from the group consisting of
saccharide syrups and
polysaccharide syrups.
It is another object of the present invention to provide the functional food
as defined above,
wherein said syrup is chosen from the group consisting of honey, date syrup,
and maple
syrup.
It is another object of the present invention to provide the functional food
as defined above,
wherein said particles are homogeneously dispersed within said food base
carrier.
It is another object of the present invention to provide the functional food
as defined above,
wherein said particles comprise microspheres.
It is another object of the present invention to provide the functional food
as defined above,
wherein said microspheres have diameters between 0.5 gm and 5 gm.
It is another object of the present invention to provide the functional food
as defined above,
wherein said particles are chosen from the group consisting of solid particles
and semisolid
particles.
It is another object of the present invention to provide the functional food
as defined above,
wherein at least 70% of said bioactive agent is associated with said
particles.
It is another object of the present invention to provide the functional food
as defined above,
wherein at least 80% of said bioactive agent is associated with said
particles.
It is another object of the present invention to provide the functional food
as defined above,
wherein between 90% and 95% of said bioactive agent is associated with said
particles.
It is another object of the present invention to provide the functional food
as defined above,
further comprising an emulsifier.
It is another object of the present invention to provide the functional food
as defined above,
wherein said emulsifier is chosen from the group consisting of PEG esters and
sucrose esters.
It is another object of the present invention to provide the functional food
as defined above,
wherein said emulsifier is a PEG ester chosen from the group consisting of
PEG6000 esters,
PEG100 stearate, and PEG40 stearate.
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It is another object of the present invention to provide the functional food
as defined above,
wherein said food base carrier is mixed with at least one additional bioactive
agent.
It is another object of the present invention to provide the functional food
as defined above,
wherein said at least one additional bioactive agent is chosen from the group
consisting of
vitamins and minerals.
It is another object of the present invention to provide the functional food
as defined above,
wherein said at least one additional bioactive agent is a hydrophilic
bioactive agent.
It is another object of the present invention to provide the functional food
as defined above,
wherein said hydrophilic bioactive agent is chosen from the group consisting
of cobalamin,
folate, and ferrous gluconate.
It is another object of the present invention to provide a method of providing
at least one
bioactive agent to a subject in need comprising administering a functional
food comprising at
least one lipophilic bioactive agent associated with lipid particles dispersed
within a food
base carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of administering a functional food further comprises a step of
administering a
functional food characterized by at least one of the following:
said lipophilic bioactive agent is chosen from the group consisting of
curcumin,
diindolylmethane, quercetin, diadezin, silymarin, genistein, essential fatty
acids, and
phytosterols; and,
said food base carrier is chosen from the group consisting of saccharide and
polysaccharide syrups.
It is another object of the present invention to provide the method as defined
above, wherein
said step of administering a functional food further comprises a step of
administering a
functional food comprising an emulsifier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of administering a functional food comprising an emulsifier further
comprises
administering a functional food comprising an emulsifier chosen from the group
consisting of
PEG esters and sucrose esters.
It is another object of the present invention to provide the method as defined
above, wherein
said step of administering a functional food further comprises a step of
administering a
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functional food comprising at least one additional bioactive agent, said at
least one additional
bioactive agent mixed with said food base carrier.
It is another object of the present invention to provide the method as defined
above, wherein
said step of administering a functional food comprising at least one
additional bioactive agent
mixed with said food base carrier further comprises a step of administering at
least one
hydrophilic bioactive agent mixed with said food base carrier.
It is another object of the present invention to provide a method of producing
functional food
comprising:
a. melting a lipid solvent including a bioactive agent at a temperature of at
least 65 C;
and,
b. emulsifying the product of step (a) with a semi-solid food base carrier
until the
formation of an emulsion comprising solid lipid particles; said solid lipid
particles
comprising said bioactive agent.
It is another object of the present invention to provide a method of producing
functional food
comprising:
a. melting a lipid solvent including a bioactive agent at a temperature of at
least 65 C;
and,
b. emulsifying (a) with a liquid food base carrier until the formation of an
emulsion
comprising solid lipid particles; said solid lipid particles comprising said
bioactive
agent.
It is another object of the present invention to provide the method as defined
above, wherein
said bioactive agent is a bioactive food component.
It is another object of the present invention to provide the method as defined
above, wherein
said bioactive food component is curcumin.
It is another object of the present invention to provide the method as defined
above, wherein
said bioactive food component is DIM.
It is another object of the present invention to provide the method as defined
above, wherein
said food base carrier is honey.
It is another object of the present invention to provide the method as defined
above, wherein
said lipid solvent includes at least one selected from a group consisting of
hydrogenated
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castor oil, stearin, palm oil, AlinaTM oil, High-omega-3 sage oil, pomegranate
oil, avocado
oil, olive oil, and any combination thereof.
It is another object of the present invention to provide the method as defined
above, wherein
said lipid solvent further includes an emulsifier.
It is another object of the present invention to provide the method as defined
above, wherein
said emulsifier includes a PEG ester or a sucrose ester.
It is another object of the present invention to provide the method as defined
above, wherein
said temperature is 70-90 C.
It is another object of the present invention to provide the method as defined
above, wherein
said food base is heated to at least 65 C prior to step (b).
It is another object of the present invention to provide the method as defined
above, wherein
a diameter of said solid lipid particles is in a range of 0.5 to 5 microns.
It is another object of the present invention to provide the method as defined
above, wherein
step (b) is effected while gradually cooling a mixture of said lipid solvent
including said
bioactive agent and said food base to a temperature of about 45 C.
It is another object of the present invention to provide a composition of
matter comprising a
food-based carrier comprising solid lipid particles homogenously dispersed
therein and at
least one bioactive agent, wherein at least 80% of said at least one bioactive
agent is
associated with said solid lipid particles.
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said bioactive agent is a bioactive food component.
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said bioactive food component is curcumin.
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said bioactive food component is Diindolylmethane (DIM).
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said food base carrier is honey.
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said solid lipid particles includes at least one selected from
a group consisting
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of hydrogenated castor oil, stearin or palm oil, AlinaTM oil, High-omega-3
sage oil,
pomegranate oil, avocado oil, olive oil, and any combination thereof.
It is another object of the present invention to provide the composition of
matter as defined
above, further comprising an emulsifier.
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said emulsifier is PEG.
It is another object of the present invention to provide the composition of
matter as defmed
above, wherein a diameter of said solid lipid particles is in a range of 0.5
to 5 microns.
It is another object of the present invention to provide a method of providing
a biaoactive
agent to a subject in need comprising administering a food-based carrier
including solid lipid
particles homogenously dispersed therein and at least one bioactive agent,
wherein at least
80% of said at least one bioactive agent is associated with said solid lipid
particles.
It is another object of the present invention to provide the method as defined
above, wherein
administering is via an oral route.
It is another object of the present invention to provide a composition of
matter comprising a
honey carrier including solid lipid particles homogenously dispersed therein
and curcumin
and/or DIM entrapped within said solid lipid particles.
It is another object of the present invention to provide the functional food
as defined above,
wherein said functional food is stable to storage at 32 C for three months.
It is another object of the present invention to provide the composition of
matter as defined
above, wherein said composition of matter is stable to storage at 32 C for
three months.
It is a further object of this invention to disclose a method for increasing
the concentration of
a lipophilic bioactive agent within the body of a patient, comprising:
incorporating said
lipophilic bioactive agent into a functional food as defined in any of the
above; and,
administering a predetermined quantity of said functional food to a patient in
need.
It is a further object of this invention to disclose a method for increasing
the concentration of
a lipophilic bioactive agent within the body of a patient, comprising:
administering to a
patient in need a predetermined quantity of a functional food as defined in
any of the above
comprising said lipophilic bioactive agent.
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It is a further object of this invention to disclose such a method for
increasing the
concentration of a lipophilic bioactive agent within the body of a patient as
defined in either
of the above, wherein said lipophilic bioactive agent is a bioactive food
component.
It is a further object of this invention to disclose such a method for
increasing the
concentration of a lipophilic bioactive agent within the body of a patient as
defined in any of
the above, wherein said lipophilic bioactive agent comprises at least one
component chosen
from the group consisting of curcumin, diindolylmethane, quercetin, diadezin,
silymarin,
genistein, essential fatty acids, and phytosterols.
It is a further object of this invention to disclose such a method for
increasing the
concentration of a lipophilic bioactive agent within the body of a patient as
defined in any of
the above, wherein said lipid solvent is chosen from the group consisting of
hydrogenated
castor oil; stearin; palm oil; high-omega-3 sage oil; pomegranate oil; avocado
oil; olive oil;
and any combination thereof.
It is a further object of this invention to disclose such a method for
increasing the
concentration of a lipophilic bioactive agent within the body of a patient as
defined in any of
the above, further comprising a step of raising the concentration of said
lipophilic bioactive
agent in the blood of said patient in need.
It is a further object of this invention to disclose such a method for
increasing the
concentration of a lipophilic bioactive agent within the body of a patient as
defined in any of
the above, further comprising a step of raising the concentration of said
lipophilic bioactive
agent in the liver of said patient in need.
It is a further object of this invention to disclose such a method for
increasing the
concentration of a lipophilic bioactive agent within the body of a patient as
defined in any of
the above, further comprising a step of raising the concentration of said
lipophilic bioactive
agent in the gastrointestinal mucosa of said patient in need.
It is a further object of this invention to disclose a method for treating a
condition ameliorated
by a lipophilic bioactive agent comprising: incorporating said lipophilic
bioactive agent into a
functional food as defined in any of the above and administering a
predetermined quantity of
said functional food to a patient suffering from said condition.
It is a further object of this invention to disclose a method for treating a
condition ameliorated
by a lipophilic bioactive agent, comprising administering to a patient in need
a predetermined
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quantity of a functional food as defined in any of the above comprising said
lipophilic
bioactive agent.
It is a further object of this invention to disclose such a method for
treating a condition
ameliorated by a lipophilic bioactive agent as defined in either of the above,
wherein said
lipophilic bioactive agent is a bioactive food component.
It is a further object of this invention to disclose such a method for
treating a condition
ameliorated by a lipophilic bioactive agent as defined in any of the above,
wherein said
lipophilic bioactive agent comprises at least one component chosen from the
group consisting
of curcumin, diindolylmethane, quercetin, diadezin, silymarin, genistein,
essential fatty acids,
and phytosterols.
It is a further object of this invention to disclose such a method for
treating a condition
ameliorated by a lipophilic bioactive agent as defmed in any of the above,
wherein said lipid
solvent is chosen from the group consisting of hydrogenated castor oil; steam;
palm oil;
high-omega-3 sage oil; pomegranate oil; avocado oil; olive oil; and any
combination thereof.
It is a further object of this invention to disclose such a method for
treating a condition
ameliorated by a lipophilic bioactive agent as defined in any of the above,
wherein said
condition is chosen from the group consisting of immune diseases, heart
disease, respiratory
diseases, inflammation, cancer, leukemia, lymphoma, gastrointestinal cancers,
genitourinary
cancers, breast cancer, ovarian cancer, head and neck squamous cell carcinoma,
lung cancer,
melanoma, neurological cancers, gastrointestinal conditions, gastric ulcers,
colitis, bowel
disease, crohn's disease, colorectal cancer, fatty liver disease and Non-
Alcoholic
Steatohepatitis (NASH), edema, arthritis, pancreatitis, ocular conditions,
inflammatory
diseases and any combination thereof.
It is a further object of this invention to disclose such a method for
treating a condition
ameliorated by a lipophilic bioactive agent as defined in any of the above,
wherein said
condition is chosen from the group consisting of respiratory papillomatosis,
prostatitis,
cataracts, allergies, bronchitis, asthma, celiac disease, non-celiac gluten
sensitivity, and
irritable bowel syndrome.
It is a further object of this invention to disclose a method for providing
adjunct therapy to a
patient suffering from cancer, comprising incorporating said lipophilic
bioactive agent into a
functional food by the method as defined in any of the above and administering
a
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predetermined quantity of said functional food to said patient in conjuction
with another
anticancer therapy.
It is a further object of this invention to disclose a method for providing
adjunct therapy to a
patient suffering from cancer, comprising administering to said patient a
predetermined
quantity of a functional food as defined in any of the above comprising said
lipophilic
bioactive agent in conjunction with another anticancer therapy.
It is a further object of this invention to disclose a method for providing
therapy in term of
Parma, i.e. botanical drug to a patient suffering from solid cancers,
leukemia, lymphoma,
gastrointestinal cancers, genitourinary cancers, breast cancer, ovarian
cancer, head and neck
squamous cell carcinoma, lung cancer, melanoma, neurological cancers, and
sarcoma, also
gastrointestinal conditions such as: gastric ulcers, colitis, bowel disease,
crohn's disease,
colorectal cancer, fatty liver disease and Non-Alcoholic Steatohepatitis
(NASH), edema,
arthritis, pancreatitis, ocular conditions, inflammatory diseases, comprising
incorporating
said lipophilic bioactive agent into a pharmaceutical drug by the method as
defmed in any of
the above.
BRIEF DESCRIPTION OF THE DRAWINGS
The invention is herein described, by way of example only, with reference to
the
accompanying drawings. With specific reference now to the drawings in detail,
it is stressed
that the particulars shown are by way of example and for purposes of
illustrative discussion
of the preferred embodiments of the present invention only, and are presented
in the cause of
providing what is believed to be the most useful and readily understood
description of the
principles and conceptual aspects of the invention. In this regard, no attempt
is made to show
structural details of the invention in more detail than is necessary for a
fundamental
understanding of the invention, the description taken with the drawings making
apparent to
those skilled in the art how the several forms of the invention may be
embodied in practice.
In the drawings:
FIG. 1 illustrates the chemical structure of curcumin and its metabolites.
FIG. 2 is a flow chart outlining a method of producing a solid lipid fine
dispersion of
curcumin (and/or DIM) in honey.
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FIG. 3 is a microscope image of curcumin powder particles (crystals) in honey
with particle
sizes of 5 - 100 m.
FIG. 4 presents microscope images of curcumin solid lipid particles (SLPs) in
honey with a
mean particle size of <5 gm; the curcumin SLPs dispersed in water; and
curcumin powder
dispersed in water.
FIGs. 5a-5c presents graphs of curcumin levels in rats following
administration of 400 mg/kg
unformulated curcumin or curcumin that had been formulated according to one
embodiment
of the present invention.
Fig. 6a illustrates the delta in the plasma curcumin concentration while using
the SLP and
Meriva as a function of time (15 minutes, 30 minutes, 60 minutes and 120
minutes).
FIG. 6b illustrates the delta in the mucosa curcumin concentration while using
the SLP and
Meriva as a function of time (15 minutes, 30 minutes, 60 minutes and 120
minutes).
DETAILED DESCRIPTION OF THE INVENTION
The following description is provided, alongside all chapters of the present
invention, so as to
enable any person skilled in the art to make use of the invention and sets
forth the best modes
contemplated by the inventor of carrying out this invention. Various
modifications, however,
is adapted to remain apparent to those skilled in the art, since the generic
principles of the
present invention have been defined specifically to provides a unique
functional food and a
method of producing the same.
Furthermore, the present invention provides a composition which can be used to
treat
disorders and promote well being. Specifically, the present invention can be
used to treat
subjects suffering from e.g., immune disorders cancer, solid cancers,
leukemia, lymphoma,
gastrointestinal cancers, genitourinary cancers, breast cancer, ovarian
cancer, head and neck
squamous cell carcinoma, lung cancer, melanoma, neurological cancers, and
sarcoma, also
gastrointestinal conditions such as: gastric ulcers, colitis, bowel disease,
crohn's disease,
colorectal cancer, fatty liver disease and Non-Alcoholic Steatohepatitis
(NASH), edema,
arthritis, pancreatitis, ocular conditions, inflammatory diseases, comprising
incorporating
said lipophilic bioactive agent into a pharmaceutical drug or any combination
thereof as will
be described hereinafer.
It is within the scope of the present invention to disclose a method for
providing therapy in
term of Parma, i.e. botanical drug to a patient suffering from solid cancers,
gastrointestinal
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conditions such as: gastric ulcers, colitis, bowel disease, crohn's disease,
colorectal cancer,
edema, arthritis, pancreatitis, ocular conditions, inflammatory diseases,
comprising
incorporating said lipophilic bioactive agent into a pharmaceutical drug by
the method as will
be disclosed hereinafter.
The principles and operation of the present invention may be better understood
with reference
to the drawings and accompanying descriptions.
Before explaining at least one embodiment of the invention in detail, it is to
be understood
that the invention is not limited in its application to the details set forth
in the following
description or exemplified by the examples. The invention is capable of other
embodiments
or of being practiced or carried out in various ways. Also, it is to be
understood that the
phraseology and terminology employed herein is for the purpose of description
and should
not be regarded as limiting.
While reducing the present invention to practice, the present inventors have
devised a novel
to approach for formulating a composition which includes a food based carrier
mixed with solid
lipid particles loaded with a bioactive agent. The present composition
facilitates delivery of
the bioactive agent to the GI tract while providing a nutrient base and
eliminating or reducing
offensive odors or tastes associated with the bioactive agent.
Thus, according to one aspect of the present invention there is provided a
composition which
can be used to treat a condition or disorder or promote or maintain well being
in a subject
such as a human. As is described hereinbelow, the present composition can be
used alone or
as a supplement to other treatment approaches.
It is a further object of this invention to disclose a method for providing
therapy in term of
Parma, i.e. botanical drug to a patient suffering from solid cancers,
gastrointestinal conditions
such as: gastric ulcers, colitis, bowel disease, crohn's disease, colorectal
cancer, edema,
arthritis, pancreatitis, ocular conditions, inflammatory diseases, comprising
incorporating
said lipophilic bioactive agent into a pharmaceutical drug by the method as
defined in any of
the above.
The present composition includes a food-based carrier mixed with solid lipid
particles loaded
with at least one bioactive agent.
As used herein, the phrase "food-based carrier" refers to any liquid or semi-
solid
composition which is defined as food or is derived from food. Examples of food-
based
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carriers include saccharide or polysaccharide syrups such as honey, date syrup
and maple
syrup. Preferred are food based carriers that naturally include a high
nutrient content as well
as beneficial elements such as minerals.
As used herein, the term "particles" when used with reference to solid lipids
refers to nano
or micro particles.
As used herein, the phrase "bioactive agent" refers to a substantially
purified substance that
exerts a local or systemic biological (e.g. physiological) effect. A bioactive
component can
include one or more bioactive agents. A bioactive component can be derived
from an extract
of a vegetable or fruit or other natural sources.
As used herein, with reference to quantities, the term "about" refers to 10
% of the nominal
quantity.
As is further described below and in the Examples section which follows, the
present
inventors have devised a novel approach for producing the present composition.
Such an
approach enables formation of the solid lipid particles (SLPs) loaded with the
bioactive agent
within the food based carrier, thus eliminating the need first to manufacture
the bioactive
agent-loaded SLPs and then to mix them with the carrier. It will be
appreciated that such an
approach substantially reduces the time and effort, as well as costs needed
for production. In
addition, such an approach obviates the need to use harmful organic solvents
such as ethanol,
hexanol, ethyl acetate, acetone, ketones or ethyl methyl ketone (see US Pat.
6,086,915).
As is described in detail in the Examples section which follows, the present
composition is
produced by co-melting lipophilic bioactive agents and lipids (that are solid
at room
temperature), heating to a temperature of between 65 and 90 C and emulsifying
the melt in
honey (or similar food based carrier) while cooling (in preferred embodiments,
to 45 C)
thereby allowing the lipids to solidify into solid or semisolid microspheres
within the honey.
The present composition is characterized by several unique features. The solid
lipid particles
are homogenously dispersed within the food-based carrier and constitute 5-40%
(w/w) of the
composition with. At least 70% (in preferred embodiments, at least 80%; in the
most
preferred embodiments, at least 90-95%) of the bioactive agent or agents are
associated with
the solid lipid particles.
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This enables as much as 30% loading of a carrier such as honey with a
bioactive agent
thereby enabling delivery of large amounts of the bioactive agent using the
present
composition.
In addition, the use of solid lipid particles for entrapping the bioactive
agent masks offensive
odors or tastes associated therewith as well as protecting the bioactive agent
from GI tract
degradation.
Furthermore, the solid lipid particles within the functional food disclosed
herein are small
and uniform in size and thus facilitate dissolution and delivery of the active
ingredient
entrapped therein as well as maintaining the smooth texture of the food
carrier. For example,
in the case of curcumin, SLP packaging of this active agent reduced the
particle size (in
water) severalfold (see Figure 4a-c).
In addition, since release of the bioactive agent from SLPs depends upon
enzymatic
degradation of the lipid, a more controlled and targeted release of the
bioactive agent is
enabled.
Finally, since SLP-carried bioactive agents are dispersed in the stomach as
very fine
micronized particles (See Figure 4b), dissolution and solubility of a
bioactive agent is
substantially enhanced by the present formulation.
As is mentioned above, several types of food based carriers are suitable for
use with the
present composition; carriers such as honey (in raw or mostly raw forms), date
syrup and
maple syrup are presently preferred due to their nutrient and mineral/element
content.
Honey
The end result of the work of the bee community is honey. It contains about
41% fructose.
This makes it the sweetest sugar known to mankind. It also contains about 35%
glucose,
about 17% water, about 2% sucrose, and small amounts of minerals and amino
acids.
Honey also contains nearly all of the trace elements that the human body
needs. The health
benefits of honey depend on its quality which is a function of the pollen
collected by honey
bees. The quality of honey is also dictated by processing which can remove
many of the
phytonutrients found in raw honey.
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Raw honey contains small amounts of the same resins found in propolis which is
a complex
mixture of resins and other substances that honeybees use to seal the hive and
make it safe
from bacteria and other micro-organisms. The resins found in propolis only
represent a small
part of the phytonutrients found in propolis and honey, other phytonutrients
found both in
honey and propolis have been shown to possess cancer-preventing and anti-tumor
properties.
These substances include caffeic acid methyl caffeate, phenylethyl caffeate,
and phenylethyl
dimethylcaffeate. Researchers have discovered that these substances prevent
colon cancer in
animals by shutting down the activity of two intracellular enzymes,
phosphatidylinositol-
specific phospholipase C and lipoxygenase. When raw honey is extensively
processed and
heated, the benefits of these phytonutrients are largely eliminated.
Date Syrup
Dates (Phoenix dactylifera L.) are an important crop in the desert regions of
Middle Eastern
countries.
Date fruit is a highly nutritious food product. It is rich in calories and in
minerals such as iron
and potassium and contains modest amounts of folate, and a small amount of
vitamins A and
B. Dates are considered beneficial for treatment of anemia, constipation and
fatigue.
Date syrup has a consistency similar to that of honey while being darker in
appearance and
having a unique flavor. Date syrup contains about 88% sugars, mainly glucose
and fructose,
and is a good source of essential elements such as calcium, phosphorus,
potassium and
magnesium. Date syrup is low in sodium, and as such it is a suitable food for
low sodium
diets.
Several bioactive agents can be used with the present composition including
for example,
curcumin, Silymarin, DIM, Genistein, Quercetin and Diadezin. Combinations of
bioactive
agents, e.g. curcumin + diindolylmethane (DIM) or essential fatty acids (EPA
and EHA) and
phytosterols such as beta sitosterol, stigmasterol, campesterol, and
brassicasterol can also be
used with the present invention.
The bioactive agent or agents are preferably associated with the solid lipid
particles (e.g.
entrapped therein) as described herein, although in some cases, the
composition can also
include additional bioactive agents (e.g. vitamins or minerals) that are
directly mixed with the
food based carrier.
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Since solid lipid particles are designed for entrapping hydrophobic (water
insoluble)
compounds, hydrophilic bioactive agents would not be suitable for SLP
entrapment and as
such are preferably added directly to the carrier of the present composition.
Thus, bioactive agents such as curcumin and DIM as well as quercetin and
diadezin which
are hydrophobic and thus poorly miscible in a food-based carrier are
preferably entrapped in
the solid lipid particles of the present composition, while other bioactive
agents such as
cobalamin, folate and iron (as ferrous gluconate) are preferably mixed
directly into the food-
based carrier of the present composition.
Curcumin
Curcumin a plant polyphenol is an edible component of the plant Curcumina
longa, a key
ingredient of Indian curry. Curcumin has been shown to be non-toxic, to have
antioxidant
activity, to inhibit the proliferation of cancer cells, and to inhibit
inflammation. Curcumin
inhibits the production of pro-inflammatory cytokines, such as TNFa,
Interferon gamma, and
IL-6 secreted by activated macrophages and T cells during an inflammatory
response. The
mechanism of action of curcumin involves, among other things, the inhibition
of important
intracellular signaling pathways that participate in cancer cells and in the
inflammatory
response mediated by NFkappaB, COX, lipooxygenase (LOX), and inducible nitric
oxide
synthase (iNOS). Curcumin is capable of reducing aberrant up-regulation of COX
and/or
iNOS in early stages of carcinogenesis and to also inhibit key intracellular
elements involved
in cancer cell proliferation. Significant preventive and/or curative effects
by curcumin have
been observed in experimental animal models of a number of diseases, including
cancer.
Because of its lack of apparent toxicity, curcumin has been recently added
(August, 2009) to
FDA's GRAS (Generally Regarded As Safe) list. The recommended maximum safe
daily
dose was set at up to 1.75 grams, per human, per day.
Ingested curcumin exhibits low plasma and tissue levels of mostly due to poor
absorption,
rapid metabolism (see Figure 1), and rapid systemic elimination (Anand et al.
Molecular
Pharmaceutics Vol. 4, No. 6; 2007). Numerous approaches have been undertaken
to improve
the bioavailability of curcumin. These approaches include use of adjuvants
such as piperine
that interfere with glucuronidation; use of liposomal or particulate delivery
approaches; use
of curcumin phospholipid complexes; and use of curcumin structural analogues
(e.g., EF-24).
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Figures 4a-c illustrate SLP-curcumin particles mixed in a honey-based carrier
(Figure 4a),
dispersed in water (Figure 4b) and curcumin crystals resultant from mixing
curcumin powder
with water (Figure 4c).
Diindolylmethane (DIM)
DIM is a bioactive agent derived from Brassica vegetables such as cabbage,
cauliflower,
Brussels sprouts and broccoli. Results of several studies indicate that DIM
exhibits promising
cancer-protective activities, especially against mammary neoplasia (breast
cancer). Oral
intubation of DIM in a single dose before carcinogen treatment reduced the
incidence and
multiplicity of dimethylbenzanthracene-induced mammary tumors in rats by 70 to
80%
(Wattenberg et al., Cancer Res., 38, (1978) 1410-1413 ). Also, repeated oral
administrations
of DIM during the promotion stage of dimethylbenzanthracene-induced mammary
tumorogenesis inhibited tumor growth in rodents by as much as 95% (Chen et
al.,
Carcinogenesis 19 (1998), pp. 1631-1639).
DIM is also widely used as adjunct therapy for recurrent respiratory
papillomatosis (Auborn
et al., Antivir. Ther. 7 (2002) 1-9.), caused by certain types of human
papillomaviruses
(HPVs).
DIM's anti-tumor activity is possibly through modulation of the immune
response. Studies
have shown that exposure to DIM could influence major immune responses,
including natural
killer cell activity, antibody production, and T-cell-mediated immunity.
Importantly, DIM
upregulates the expression of both interferon (IFN)-y and IFN-y receptor, and
potentiates the
effects of IFN-y-induced expression of MHC I antigens in human breast cancer
cells, making
them susceptible to effector T cells.
IFN-y is a central regulator of immune and inflammatory responses that
contributes to the
inhibition of primary and transplanted tumor development, as well as anti
viral response.
Thus, if DIM has generalized immune stimulatory properties, these capacities
might indeed
contribute to its anti-carcinogenic effects. In mice, DIM was shown to induce
splenocyte (B
and T cell) proliferation, reactive oxygen species (ROS) generation, cytokine
production and
resistance to viral infection (Xue et al., J. Nutr. Biochem. 19, (2008), 336-
344). The addition
of DIM to cultured cells enhanced both splenocyte proliferation and ROS
production by
peritoneal macrophages. Since cytokines are major mediators of host defense
they regulate
communication between antigen-presenting cells, lymphocytes and other host
cells during an
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immune response. The cytokine repertoire present at a tissue site determines
the type of host
response directed against a tumor or an infection. Indeed, cytokines promoting
the
development of T-cell mediated immunity can induce or enhance the anti-tumor
and anti-
microbial immunity. Importantly, the hematopoietic growth factor, G-CSF is a
cytokine that
induces the bone marrow to generate more leukocytes, which are essential for
fighting
infection and cancer. Collectively, these data strongly suggest that DIM has
potent
immunomodulating activities that are consistent with the anti-tumor, anti-
viral and anti-
bacterial activities of this dietary Indole.
Quercetin
Quercetin is a plant-derived flavonoid that may have anti-inflammatory and
antioxidant
properties. Quercetin may have positive effects in combating or helping to
prevent cancer,
prostatitis, heart disease, cataracts, allergies/inflammations, and
respiratory diseases such as
bronchitis and asthma. An 8-year study found that the presence of the three
flavonols
kaempferol, quercetin, and myricetin in a person's normal diet was associated
with a reduced
risk of pancreatic cancer. Quercetin has demonstrated significant anti-
inflammatory activity
by inhibiting both manufacture and release of histamine and other
allergic/inflammatory
mediators. In addition, it exerts potent antioxidant activity and vitamin C-
sparing action. In
mice, an oral quercetin dose of 12.5 to 25 mg/kg increased gene expression of
mitochondrial
biomarkers and improved exercise endurance. An in vitro study showed that the
combination
of quercetin and resveratrol inhibited production of fat cells.
Genistein/Diadezin
Genistein and diadzein are isoflavones found in a number of plants including
lupin, fava
beans, soybeans, kudzu, and psoralea. Besides functioning as antioxidants and
anthelmintics,
many isoflavones have been shown to interact with animal and human estrogen
receptors,
causing effects in the body similar to those caused by the hormone estrogen.
Genistein and other isoflavones have been found to have antiangiogenic effects
(blocking
formation of new blood vessels), and may block the uncontrolled cell growth
associated with
cancer, most likely by inhibiting the activity of substances in the body that
regulate cell
division and cell survival. Studies have also found genistein to be useful in
combating
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leukemia and that it can be used in combination with certain other leukemia
combating drugs
to improve their efficacy.
Silymarin
Silymarin (SM) is a lipophilic extract of milk thistle and is composed of
several isomer
flavonolignans. In vitro and animal research suggest that silibinin (major
active component of
silymarin) has hepatoprotective (antihepatotoxic) properties and anti-cancer
effects in human
prostate adenocarcinoma cells, estrogen-dependent and -independent human
breast carcinoma
cells, human ectocervical carcinoma cells, human colon cancer cells, and both
small and
nonsmall human lung carcinoma cells; see, for example,
http ://en.wikipedia.org/wiki/S ilibinin#cite_note-0.
Cobalamin (Vitamin B12)
Cobalamin contributes to the formation of red blood cells and bone marrow, the
metabolism
of carbohydrates, fats and proteins, nerve and cardiovascular functions and
plays a role in
DNA synthesis. Deficiency of this vitamin may cause anemia, exhaustion,
irritation,
depression, shortness of breath, difficulty walking, memory loss, mood swings,
disorientation, dementia and constipation. Cobalamin doses greater than 3
milligrams daily
may cause eye conditions. Cobalamin deficiency occurs primarily in people with
malabsorption and myelodysplastic syndrome. The remaining cases of nutrient-
deficiency
anemia are usually associated with cobalamin, most frequently related to food-
cobalamin
malabsorption, and/or Folate deficiency. Cobalamin deficiency occurs
frequently among
patients with malabsorption, but it is often unrecognized or not investigated
because the
clinical manifestations are subtle.
Folate
Folate plays an important role in the metabolism of nucleic acids and amino
acids.
Consequentially, it is essential for cell growth and development and normal
functionality of
the nervous system. Folate deficiency in the elderly is characterized by
anemia which is
accompanied by symptoms such as, shortness of breath, fatigue and weakness.
Folate
deficiency may also cause a sore tongue, depression, nerve damage and infant
neural tube
defects, heart defects and limb malformations. Folate doses larger than 400
micrograms daily
may cause anemia and may mask symptoms of a cobalamin deficiency.
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Iron
Iron is an essential part of many biological molecules such as hemoglobin. The
human body
absorbs iron from animal sources faster than iron from plant sources. Iron
deficiency is a
common condition and can lead to disorders such as anemia. The recommended
daily dose of
iron for humans is about 10 milligrams; a normal diet typically provides that
amount but
patients with malabsorption or frequent diarrhea require supplementation.
As is mentioned above, the present composition can be used to treat disorders
or promote
good health.
Thus, according to another aspect of the present invention there is provided a
method of
treating a subject having a disorder which would benefit from the
physiological effects of the
curcumin and/or DIM provided by the present composition and the nutritional
effects of the
food based carrier of the composition.
One example of such a disorder is cancer. In cancer treatment, the food based
carrier is
honey in its minimally processed form, since it contains natural anti-cancer
chemicals and
provides nutrition and the physiologically active ingredients are DIM and
optionally
curcumin (both in SLPs).
Such a composition is especially suitable for use as adjunct cancer therapy.
The effect of
chemotherapy is not specific to cancer cells and as such, it leads to severe
side effects, such
as immunosuppression and myelosuppression. DIM would reduce chemotherapy-
induced
granulocytopenia, and stimulate immune responses in chemotherapy-treated
cancer patients
that are immunocompromised. In addition, since honey, DIM and curcumin have
anti-viral
and anti-bacterial properties, consumption of the present composition by
chemotherapy-
treated cancer patients, can help reduce the incidence of viral and bacterial
infections
associated with chemotherapy.
Cancer patients can be treated with three daily doses of the present
composition, with each
dose including 1 gram of honey, about 200 mg of DIM and about 400 mg of
curcumin.
The present invention can also be used to treat disorders associated with
aging and
immunosenescence.
The immune system undergoes characteristic and multifaceted changes with
aging. These
changes occur in all type of leukocytes, including, neutrophils, T-cells, B-
cells,
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monocyte/macrophages, dendritic cells and natural killer cells. Accordingly,
aging affects
both innate and adaptive immune functions. Collectively, this process is
called
"immunosenescence" and it refers to the natural gradual deterioration of the
immune system
with advancing age. It involves both the host's capacity to respond to
infections and the
development of long-term immune memory, especially by vaccination. This age-
associated
immune deficiency is ubiquitous and found in both long- and short-lived
species as a function
of their age relative to life expectancy rather than chronological time. It is
considered a major
contributory factor to the increased frequency of morbidity and mortality
among the elderly.
Immunosenescence is not a random deteriorative phenomenon, rather it appears
to inversely
to repeat an evolutionary pattern and most of the parameters affected by
immunosenescence
appear to be under genetic control. hnmunosenescence can also be sometimes
envisaged as
the result of the continuous challenge of the unavoidable exposure to a
variety of antigens
such as viruses and bacteria. Immunosenescence is a multifactorial condition
leading to many
pathologically significant health problems in the aged population. Age-
dependent biological
changes such as depletion of hematopoietic stem cells, decline in the total
number of
phagocytes and NK cells and a decline in humoral immunity contribute to the
onset of
immunosenescence.
Thus aging is associated with a decline in both the production of new naive
lymphocytes and
the functional competence of memory cell populations. This has been implicated
in the
increasing frequency and severity of diseases such as cancer, chronic
inflammatory disorders
and autoimmunity. A problem of infections in the elderly is that they
frequently present with
non-specific signs and symptoms. Ultimately, this provides problems in
diagnosis and
subsequently, treatment.
Anemia is highly prevalent in the elderly population, including those
individuals who are in
long term care facilities and geriatric wards. Importantly, even mild anemia
is associated with
adverse health outcomes. Therefore, anemia is of a great healthcare concern.
The most
frequent etiologies of anemia in the elderly are anemia of chronic
disease/inflammation; iron,
folate and vitamin B12 (cobalamin).
A composition including honey, DIM and vitamins can be used to treat viral and
bacterial
infections as well as anemia in populations such as the elderly, the
malnourished and the like.
The honey and DIM would provide antibacterial and antiviral functions while
DIM would
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increase hematopoietic cytokine production. Key vitamins such as cobalamin and
folate will
enhance blood production.
Treatment can include five daily doses of a composition including 1 g of
honey, 200 mg of
DIM, 50 lig of cobalamin, 20 pg of folate and 3 mg of iron.
The present invention can also be used to treat disorders such as Celiac
Disease (CD), Non-
Celiac Gluten Sensitivity (NCGS) and Irritable Bowel Syndrome (IBS).
Celiac disease is an immunologically-mediated enteropathy, triggered in
genetically
susceptible subjects, many of whom carry the HLA-DQ (haplotypes 2 or 8). The
intake of
certain proteins, primarily gluten from wheat, and similar proteins from
barley and rye induce
the disease. Up to 1% of the entire population suffers from CD. People with CD
also have
one or more additional food allergies, which may include milk protein, corn
and soy.
Symptoms include chronic diarrhea, abdominal pain, cramping, weight loss or
growth
inhibition, musculoskeletal pain, neurological involvement and fatigue.
There is evidence suggesting that upon exposure to gluten an immune reaction
is triggered
towards gluten-derived peptides generated by the enzyme transglutaminase. In
turn, these
gluten-derived peptides stimulate immune cross-reaction against self
components of the small
bowel. This classical autoimmune response gives rise to an inflammatory
reaction in the
intestines. CD causes small-bowel mucosal villous atrophy with crypt
hyperplasia.
Importantly, the disease significantly increases the risk of adenocarcinoma of
the small
intestines and lymphoma of the small bowel.
The inflammatory process, mediated predominantly by T cells, leads to
disruption of the
structure and function of the small bowel's mucosal lining and causes
malabsorption; it
impairs the body's ability to absorb nutrients, minerals such as calcium and
iron, and fat-
soluble vitamins. In fact, CD can be considered as an archetypal malabsorption
syndrome,
and is a frequent cause of anemia. In many cases anemia has been reported as
the only initial
manifestation, or the most frequent extra-intestinal symptom of CD. Folate
(vitamin B9) and
cobalamin (vitamin B12) deficiencies are known complications of CD. Another
common
nutritional anemia associated with CD is iron deficiency. Iron deficiency
anemia was
reported in up to 46% of patients with subclinical CD. Some CD patients also
develop
osteoporosis and bone fractures due to the malabsorption of vitamin D and
hypocalcaemia.
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Gluten intolerance is a broad term which includes various degrees of gluten
sensitivity. It is
estimated that about 15% of all people are gluten sensitive, but only about 1%
have CD as
determined by testing positive in the currently available immunoassays and
biopsies.
Although antibody serology is an important tool in the confirmation of CD, it
does not
always correlate with the typical mucosal appearance of CD, namely, the villus
atrophy of the
small intestine. Some patients with positive CD serology have normal
intestinal mucosa
without full blown symptoms of CD. However, they are at increased risk of
future CD
development.
The majority of people with gluten sensitivity do not have CD. Rather, they
have a condition
called Non Celiac Gluten Sensitivity (NCGS). People with NCGS exhibit
gastrointestinal
symptoms including, diarrhea, bloatdeness, cramping, etc. Also associated with
NCGS are
headache, fatigue, infertility, miscarriage and malabsorption. It is believed
that, like in CD,
the immune response of people with NCGS is triggered by gluten, but it does
not lead to the
destruction and remodeling of the intestines. A recent publication suggests
that, even in the
absence of fully developed CD, gluten can induce symptoms similar to irritable
bowel
syndrome (IBS). That study presented the hypothesis that gluten sensitivity
and post-
infectious IBS provide two triggers that can explain at least part of the
spectrum that
constitutes IBS.
IBS affects between 5-20% of the population of the western world. The wide
range may be
linked to the fact that the symptoms of non-specific dyspepsia are sometimes
confused with
those of IBS. Importantly, IBS is the second leading cause of work
absenteeism. Women are
four times more likely to suffer from IBS than men. Symptoms of IBS include
irregular
bowel function, bloating, abdominal pain, cramping, diarrhea, and
constipation. Stress and
other psychological factors are known to exacerbate the intensity and
frequency of the
syndrome.
The main characteristic of IBS is irregular defecation, and IBS is divided
into two types
based on the irregularity observed: IBS-D (diarrhea) and IBS-C (constipation).
It is estimated
that about 50% of all IBS patients use alternative medicine because of the
lack of
effectiveness of currently prescribed medications.
Recent scientific data indicate that inflammation plays an important role in
the
pathophysiology of IBS. IBS patients have a significantly elevated level of
the pro-
inflammatory cytokine IL-6 in their circulation, as well as a very high level
of the
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cyclooxygenase (COX) cycle metabolite prostaglandin E2, which is known to be
produced by
inflammatory stimuli. Evidence also indicates that the inflammatory response
is the result of
immune activation. The current scientific working hypothesis for the syndrome
is that there is
a link between the central nerve system and immune activation of unknown
etiology which
interacts and leads to gastrointestinal inflammation.
At present, the only effective treatment of CD, and NCGS is a life-long gluten
free diet. No
medication exists that will prevent the damage or prevent the
immune/inflammatory system
from attacking the gut when gluten is present. With the exception of CD
patients who became
refractory to gluten-free diet, strict adherence to the gluten-free diet
allows the intestines to
to heal, leading to resolution of all symptoms in most cases. Also, depending
on how soon the
gluten-free diet has begun, the increased risk of osteoporosis and the
development of
intestinal cancer are reduced. In many countries, gluten-free products are
available by
physician's prescription and are reimbursed by health insurance plans. The
diet, however,
can be cumbersome; failure to comply with the diet causes the severe relapse
of symptoms.
Gluten-free products are usually more expensive and harder to find than common
gluten-
containing foods. The total market of gluten-free products in the USA is about
$2 billion a
year.
The common pathophysiological manifestation of CD, NCGS and IBS is intestinal
inflammation. Thus, a composition including a food based carrier and curcumin
in solid lipid
particles can be used to treat such disorders. Curcumin inhibits stimulus-
dependent activation
of immune cells and the production of pro-immune/inflammatory cytokines
without causing
the severe side-effects frequently associated with the use of anti-
inflammatories that inhibit
the COX and LOX cycles.
It should be noted that curcumin has a chemopreventive effect against the
development of
cancer. Therefore, its consumption via honey, on a regular basis, may reduce
the high risk of
adenocarcinoma and lymphoma development in the intestines of people with CD,
as well
other patients' populations suffering from intestinal malfunction.
Treatment can include three daily doses of a composition that includes 1 gram
of the honey
and about 400 mg of curcumin (in SLPs).
The present compositions can be presented in a dispenser device, such as a
squeeze bottle or
foil sachet or encapsulated in ingestible capsule (e.g. gel capsule) which may
be packaged in
foil blisters. The dispenser device or pack may include one or more dosage
units of the
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present composition. The dispenser device or capsule pack may be accompanied
by
instructions for consumption. The dispenser device or pack may also be
accompanied by a
notice in a form prescribed by a governmental agency regulating the
manufacture, use, or sale
of neutraceuticals, which notice is reflective of approval by the agency of
the form of the
compositions for human or veterinary administration. Such notice, for example,
may include
labeling approved by the U.S. Food and Drug Administration.
Additional objects, advantages, and novel features of the present invention
will become
apparent to one ordinarily skilled in the art upon examination of the
following examples,
which are not intended to be limiting. Additionally, each of the various
embodiments and
aspects of the present invention as delineated hereinabove and as claimed in
the claims
section below finds experimental support in the following examples.
EXAMPLES
Reference is now made to the following examples, which together with the above
descriptions, illustrate the invention in a non limiting fashion.
Solid Lipid Micronized Dispersions of Curcumin in Honey
A solid lipid dispersion is a delivery system in which the bioactive agent is
solubilized or
dissolved in hot melted lipids that do solidify at room temperature. The hot
melt is emulsified
with an aqueous medium and cooled to room temperature to solidify as particles
which
include the bioactive agent within their core.
The present study tested whether curcumin, a polyphenolic compound derived
from dietary
spice turmeric, can be emulsified within a honey-based carrier using Solid
Lipid Dispersion
technology.
In order to optimize the solid lipid emulsion and enhance its curcumin-
carrying capability,
the present inventors conducted several pre-formulation studies which included
the
following:
(i) testing solubility of curcumin in various oils;
(ii) design of solid lipid formulation matrixes for curcumin in honey;
(iii) testing formulations properties including consistency and organoleptic,
taste and odor
properties; and
(iv) testing the formulation in a pilot process for physical and chemical
stability.
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A general list of components used in the present study is provided in Table 1
below.
Table 1: materials
4141,7ai NneZ--74 7-Synopyrn
Cocoglycerids Novata A
Glyceryl Monostearate GMS V PH Cutina GMS V PH
Mono/Diglycerids of cap rylic acid Capmul MCM C8
Peg-40 stearate Mirj 52s
Polyethylene glycol 100 stea rate Myrj 59P
Polyethylene glycol 400 PEG 400
Polysorbate 80 Montanox 80 G PfTwe
Polyethylene Glycol 6000 PEG 6000
Sucrose Fatty Acid 1811 Surfope 1811
Sucrose Fatty Ester Surfope 1815
Hydrogenated castor oil Cutina HR PH
Capric/Caprilic
MCT oil Triglycerides
Honey
3,3- Diindolylmethane DIM
Curcumin C3 complex Curcumin
EXAMPLE 1
Curcumin Solubility
Curcumin is insoluble in water and therefore its solubility was tested in
various solvents.
Curcumin solubility was measured by vigorously mixing 400 mg of curcumin in 2
g of a test
solvent and heating the sample to 90 C. Samples were cooled to room
temperature and
inspected visually and solubility recorded. Table 2 below provides the results
of the
solubility study. Curcumin showed very limited or poor solubility in the oils
tested, but was
soluble in Polyethylene glycol (PEG).
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Table 2: Curcumin solubility
Curcumin solubility 0.49 Curcumin in 2gr Solvent
Solvent Solubility
Capmul MCM C8 partly soluble, recrystallizes
at low temp
Propylene Glycol Monocaprylate Capryol 90 HLB6 not soluble
MCI oil not soluble
Glycerin not soluble
PEG 400 soluble
Honey room temperature not soluble
Honey 50C not soluble
crushed sesame seeds - 0.4g curcumin in 7g not soluble, dispersive
olive oil not soluble
glycerin mono oleate not soluble
Span 60 not soluble
Triomal (Omega3 and 6)a not soluble
crushed sesame seeds and honey not soluble, dispersive,
colors tong yellow
In order to further test curcumin solubility, a solubility matrix was
constructed (Table 3).
The solubility of curcumin in various lipid mixtures was tested and the
mixture was then
homogenized in honey.
Table 3: Curcumin solubility matrix
CUR1 CUR2 CUR3 CUR4 CURS CUR6 CUR7 CUR8
WAN WAN ,WAN WAN WAN WAN WAN WAN
Curcumin 33.33 33.33 33.33 33.33 33.33 33.33 33.33
33.33
Hydrogenated Castor oil 33.33 33.33
Caprylic/Caprictriglycerids 33.33 33.33
33.33
Cocoglycerids 33.33 33.33
33.33
PEG 100 stearate 33.33 33.33 33.33
Sucrose Ester 1815 33.33 33.33
Polysorbate 80 33.33
33.33 33.33
Total 99.99 99.99 99.99 99.99 99.99 99.99 99.99
99.99
Mx tem p 100.00 100.00 100.00 100.00 100.00 100.00
100.00 100.00
Solubility partly no no partly no partly
partly partly
Crystals Yes in SLN yes yes yes yes yes
yes yes
State solid two phases semi solid solid solid solid
two phases semi solid
1
Using the solubility matrix of Table 3, the present inventors discovered that
curcumin
solubility or partial solubility correlates with the presence of PEG and that
PEG therefore
contributes to curcumin solubility. Microscopic inspection of curcumin in
hydrogenated
castor oil (Cutina FIR) with PEG 100 stearate produced curcumin as solid
lipids. This
combination was then selected as the initial lipid phase of the curcumin-honey
formulation.
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EXAMPLE 2
Development of a Solid Lipid Curcumin in Honey Formulation
'Hot melt emulsification' procedures were developed and tested with respect to
production
of very fine micronized SLP well dispersed in honey. Procedures were optimized
to
maximize entrapment of curcumin in the SLPs and minimize crystallization or
precipitation
of free curcumin in honey. In addition, parameters related to taste, smell and
texture of the
honey carrier were also monitored. The general production process is outlined
in Figure 2.
Procedure A: The lipid phase (curcumin in hydrogenated castor oil with PEG 100
stearate)
was heated to 90 C and the honey was heated to 70 C. The heated lipid and
honey phases
were mixed and homogenized using Heidolph DIAX 900 homogenizer at speed 3
until the
temperature dropped to at least 20 C below the emulsification temperature and
the
formulation appeared semi solid.
Procedure B: Same as procedure A, but a portion of the honey is not heated
above 50 C in
order to preserve unique honey properties that are destroyed above 55C. The
lipid phase was
heated to 90 C; half the honey was heated to 70 C and the rest to 40 C - 50
C. The lipid
and the hot honey (70 C) phases were mixed and homogenized with Heidolph
homogenizer
DIAX 900 at speed 3 until the temperature dropped below 60 C. The rest of the
honey was
then heated to 50 C and added while vigorously mixing and/or homogenizing to
the Lipid-
honey mix until the formulation reached homogeneity.
The formulations produced by the present study were visually inspected for
homogeneity,
observed under a microscope, tested for smell taste and texture, and for
physical and
chemical stability.
Table 5 summarizes the results of three curcumin in honey formulations, CUR13,
14 and 15.
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Table 5: Hydrogenated Castor oil-PEG 100 Stearate Formulations
CUR13 CUR14 CUR15
WNV% WAN WAN
Curcumin 11.11 5.50 0.00
Hydroginated Castor oil 11.11 5.50 11.11
PEG 100 stearate 11.11 5.50 11.11
Hot Honey (70C) 66.67 33.33 75.00
Cold Honey (40-50C) 0.00 50.00 0.00
Total 100.00 99.83 97.22
Production:
Production process A B A
Solubility in oil phase yes yes yes
Lipid phase temp (process B) 100C 90 90
Tests ZT:
Crystals Few and small Few and small Few amorphous
State semi solid semi solid semi solid
Smell Honey Honey Honey
Nixing with water good good good
Taste Honey Honey Honey
CUR13 and CUR15 were produced using procedure A, while CUR14 was produced
using
procedure B. Hydrogenated castor oil was the lipid base and PEG100 stearate
was used as an
emulsifier. CUR15 was a control formulation used for examining the effect of
lipid phase
concentration and the production procedure on particle size. This formulation
was
microscopically compared to CUR13 and CUR14.
Following these tests, the present inventors concluded that PEG100 stearate is
a good
stabilizer and curcumin solvent, but that the mixture solidifies at a
relatively high temperature
(above 70 C), making it difficult to carry out the preparation procedure and
thus encouraging
the present inventors to seek alternative stabilizing emulsifiers.
Table 6: lecithin Formulation
Lecitin 5.698005698
Sucrose Ester 1811
Glyceryl Monostearate 5.698005698
Hot Honey (70C) 38.46153846
Cold Honey (40-50C) 38.46153846
Total 100
Production:
Production process
Solubility in oil phase no
Lipid phase temp (process B) 110
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PEG 100 Stearate was replaced with PEG 40 Stearate which has a lower melting
point
(CUR17, Table 7). This formulation was produced using procedure A and
exhibited
satisfactory results. Processing was easier due to lower temperature
solidification and the
resulting formulation was characterized by good stability and solubility. It
had a typical
honey smell. The observation of a small number of curcumin crystals indicates
that the
curcumin was not completely solubilized. The amount of solubilization was not
quantified,
but it is estimated that the fraction of curcumin that remained unsolubilized
was between 1 to
10%.
Table 7: PEG 40 Stearate Formulation
CUR17
Ingredients %WM
Curcumin 7.14
Hydroginated Custor oil 7.14
PEG 40 stearate 7.14
Hot Honey (70C) 78.57
Cold Honey (40-50C) 0.00
Total 100.00
Production:
Production process A
Solubility in oil phase partly
)Homogenization tern perture (process A 80
Tests ZT:
Crystals few
State semi solid
Smell honey
Nixing with water poor
In the formulation of Table 8, the concentration of PEG 40 stearate was
reduced and PEG
6000 was added to the lipid phase. The formulation was produced using
procedure A. The
result was a honey smelling formulation with very few detectable crystals and
good water
miscibility of the product and homogeneous dispersion of the SLP in water
following honey
dissolution. Particles are typically 0.5 ¨ 5 gm in diameter, indicating that
they are capable of
being dispersed efficiently in gastric fluid.
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Table 8: PEG 6000 Formulation
CUR18
Ingredients YoW/W
Curcumin 7.14
Hydroginated Custor oil 7.14
Polyethylene Glycol 6000 4.29
PEG 40 stea rate 2.86
Hot Honey (70C) 78.57
Cold Honey (40-50C) 0.00
Total 100.00
Production:
Production process A
Solubility in oil phase partly
)Homogenization tern perture (process A 80
Tests Zr:
Crystals few
State semi solid
Smell honey
Nixing with water very good
In the formulations of Table 9, a sucrose ester component was tested as the
sole stabilizing
emulsifier for curcumin using procedure A. The resulting formulation included
fine
micronized solid lipid particles exhibiting poor water miscibility, probably
due to a lack of
hydrophilic components such as the PEG moiety of the PEG stearate component.
Table 9: Sucrose Ester Formulation
CUR1 9
Ingredients VoVV/VV
Curcumin 7.14
Hydroginated Custor oil 7.14
Polyethylene Glycol 6000 4.29
Sucrose Ester 1811 2.86
Hot Honey (70C) 78.57
Cold Honey (40-50C) 0.00
Total 100.00
Production:
Production process A
Solubility in oil phase partly
)Homogenization tern perture (process A 80
Tests Zr:
Crystals few
State semi solid
Smell honey
Mixing with water none
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CUR 20 and CUR21 (Table 10) were produced using procedure A. CUR20 contained
sucrose
ester with PEG40 stearate, while that of CUR21 contained sucrose ester with
Tween 80 as
stabilizing surfactants. Both were well miscible in water; in the CUR21
formulation the
typical smell of honey was lost.
Table 10: Sucrose Ester and PEG 40 Stearate or Tween 80 Formulations
CUR20 CUR21
Ingredients %W/W %W/W
Curcumin 7.14 7.14
Hydrogenated Castor oil 7.14 7.14
Polyethylene Glycol 6000 4.29 4.29
PEG 40 stearate 1.00 0.00
Tween 80 0.00 1.00
Sucrose Ester 1811 1.00 1.00
Hot Honey (70C) 78.57 78.57
Cold Honey (40-50C) 0.00 0.00
Total 99.14 100.14
Production:
Production process A A
Solubility in oil phase partly partly
Homogenization temperture (process A) 80 80
Tests ZT:
Crystals few few
State semi solid semi solid
Smell honey less then honey
Mixing with water good good
particle size 0-6micron 0-6micron
CUR22 was produced using procedure A while CUR 22(2) was produced using
procedure B
(Table 11). The lipid phase was identical for both formulations, and included
0.5% sucrose
ester and 1.0% PEG40 stearate. Both were miscible in water, and both exhibited
a typical
honey smell and a curcumin aftertaste.
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Table 11: Optimization of Sucrose Ester concentration and production procedure
CUR22 CUR22(2)
Ingredients %WM
Curcumin 7.14 7.14
Hy:Iroginated Custor oil 7.14 7.14
Polyethylene Glycol 6000 4.29 4.29
PEG 40 stea rate 1.00 1.00
Sucrose Ester 1811 0.50 0.50
Hot Honey (70C) 78.57 39.20
Cold Honey (40-50C) 0.00 39.20
Total 98.64 98.47
Production:
Production process A
Solubility in oil phase partly partly
Homogenization temperature (process A) 80 80
Tests Zr:
Crystals almost none Few
State semi solid semi solid
CUR23 (Table 12) was produced using procedure A or B. The lipid phase was
identical to the
CUR22 formulation but 4% curcumin was used. The result was a smooth semisolid
formulation with typical honey smell and no aftertaste.
Table 12: 4% Curcumin Formulation
CUR23
Ingredients %WM
Curcumin 4.00
Hydrogenated Castor oil 7.14
Polyethylene Glycol 6000 4.29
PEG 40 stearate 1.00
Sucrose Ester 1811 0.50
Hot Honey (70C) 78.57
Cold Honey (40-50C) 0.00
Total 95.50
Production:
Production process A
Solubility in oil phase partly
Homogenization tem perture (process A) 70
Tests Zr:
Crystals wry little
State semi solid
CUR24 (Table 13) was produced with food grade stearin as the lipid solvent.
CUR25 (Table
13) was produced with food grade palm oil as the lipid solvent. Both
formulations were
produced using procedure A and resulted in a formulation having a smooth
texture, a honey
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smell and no aftertaste. The microscopic observations showed solid lipid
spherical particles
loaded with curcumin and very few free curcumin crystals. The diameter of the
solid lipid
spheres was between 0.5 and 5.0 gm.
Table 13: Stearin of Palm oil (as lipid solvent) formulations
CUR24 CUR25
Ingredients W/W W/VV
Curcum in 4.00 4.00
Stearin 7.00 0.00
Palm oil 0.00 7.00
Polyethylene Glycol 6000 4.00 4.00
PEG 40 stea rate 1.00 1.00
Sucrose Ester 1811 0.50 0.50
Hot Honey (70C) 83.50 83.50
Cold Honey (40-50C) 0.00 0.00
Total 100.00 100.00
Production:
Production process A A
Solubility in oil phase partly partly
)Homogenization tem perture (process A 70C 70C
Tests ZT:
Crystals few few
State semi solid semi solid
Smell honey honey
CUR26 is the same formulation as CUR24 but without PEG6000; CUR27 is the same
formulation as CUR25 but without PEG6000 (Table 14). The purpose of these
formulations
was to determine whether PEG6000 was necessary as the curcumin solvent. Both
formulations were produced using procedure A and in both lipid phases no
curcumin
solubility was observed. Microscopic observation of these formulations showed
spherical
lipid particles with no curcumin entrapment and curcumin crystals of various
size and
unusual shapes. These formulations did not exhibit a homogeneous texture. Both
formulations were characterized by a honey smell and a good taste.
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Table 14: Formulations without PEG 6000
CUR26 CUR27
Ingredients WM WM
Curcumin 4.00 4.00
Stearin 7.00 0.00
Palm oil 0.00 7.00
Polyethylene Glycol 6000 0.00 0.00
PEG 40 stearate 1.00 1.00
Sucrose Ester 1811 0.50 0.50
Hot Honey (70C) 87.50 87.50
Cold Honey (40-50C) 0.00 0.00
Total 100.00 100.00
Production:
Production process A A
Solubility in oil phase no no
)Homogenization temperture (process A 70C 70C
Temp at the end of production 55C 55C
Prod Homogeneity yes yes
Tests Zr:
Crystals yes yes
State semi solid semi solid
Smell
In addition to the above specific examples, the present approach can also be
used to generate
DIM, Genistein, Diadzin or Quercetin SLP in honey formulations. Table 15 below
provides
a list of ingredients that can be used in such formulations.
Table 15
Formulation 131M1 DIM2 DIM3 DIM4 GEN/DI QUE
Ingredients %W/W %W/W %W/W %W/W %W/W %W/W
Diindolylmethane 2 2 3 3 --
--
Genistein or Diadezin -- -- -- -- 1
--
Quercetin -- -- -- -- --
1
Stearin 10 8 10 8 12
12
Sucrose ester HLB 6 2 2 2 2 2
2
Tween 80 1 0.5 1 0.5 0.5
0.5
PEG 6000 -- 8 -- 8 --
--
Honey To 100 To 100 To 100 To 100 To 100 To 100
EXAMPLE 3
CUR64 (Table 16 below) was produced with AlmaTM oil (omega-3 rich sage oil).
CUR64 (Table 16 below) was produced using procedure A and resulted in a
formulation
having a smooth texture, a semisolid product with no smell, a honey sweet
taste, and no
aftertaste.
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Table 16: production with AlinaTM oil (sage omega3 rich oil)
Ingredients CUR64 - production with AlmaTM oil
W/W
Curcumin Curcumin 4
Alina oil 5
Polyethylene Glycol
6000 PEG 6000 6
Sucrose Ester HLB 15 Sisterna SP70 1
Polysorbate 80 Tween 80 0.5
Plurol Oleique
Polyglycery1-3-Dioleate 497 1
Cold Honey 51
Date syrup hot Tamar Kineret 31.5
SUM 100
Tests ZT:
Crystals mostly small, some 25micron
mostly sub-micron, with few particles up to
Emulsification 6micron
State semi solid
Smell good
Mixing with water good
taste sweet
texture smooth
EXAMPLE 4
CUR65, CUR66 and CUR67 (Table 17) were produced with pomegranate oil (CUR65),
with
avocado oil (CUR66) and with olive oil (CUR67).
CUR65, CUR66 and CUR67 (Table 17) were produced using procedure A and resulted
in a
smooth semisolid product having a smooth texture, no smell, a sweet taste, and
no aftertaste.
In all three formulations (CUR65, CUR66 and CUR67) microscopic observation
revealed a
few 10-25 jim curcumin crystals and lipid particles up to 3 [un in diameter.
Table 17: production with Pomegranate oil (CUR65), with Avocado oil (CUR66)
and with
Olive oil (CUR67)
production with production with production with
pomegranate oil avocado oil olive oil
CUR65 CUR66 CUR67
%W/W %W/W %W/W
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Curcumin 4 4 4
Pomegranate seed oil 5
Avocado oil 5
Olive oil 5
Polyethylene Glycol
6000 6 6 6
Sucrose Ester HLB
15 1 1 1
Polysorbate 80 0.5 0.5
0.5
Polyglycery1-3-
Dioleate 1 1 1
Cold Honey 51 51 51
Date syrup hot 31.5 31.5
31.5
SUM 100 100
100
Tests ZT:
Crystals some 10-25micron some 10-25micron
some 10-25micron
mostly sub-lmicron, some submicron to submicron to
Emulsification 3micron 3micron
2micron
State semi solid semi solid
semi solid
Smell Typical pomegranate oil good
good
Mixing with water good good
good
taste sweet with typical smell sweet
sweet
texture smooth smooth
smooth
EXAMPLE 5
The ingredients of CUR66-1, CUR66-2 and CUR66-3 ingredients are given in Table
18.
CUR66-1 was produced as follows (production of 1 kg):
a. Curcumin was melted with PEG6000 to 90 C;
b. Lipid phase was added to the PEG-curcumin solution and heated with stirring
to 80
or ;
c. 90% of the date syrup was heated to 75 C; since 10% was used to solubilize
the
microbial preservative potassium sorbate
d. 10% of the date syrup was heated to 70 C with potassium sorbate,
solubilised and
cooled to room temperature;
e. The mixture was homogenized with "Silverson L4RT emulsor screens";
f. The lipid phase was homogenized at 5000 rpm and date syrup was added slowly
with
homogenization for 3 min;
g. The homogenization speed was reduced to 4000 rpm for 4 min with cooling in
a water
bath;
h. Honey and potassium sorbate were added at room temperature;
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i. The mixture was mixed manually.
CUR66-2 was produced as follows (production of 1.5 kg):
a. Curcumin was melted with PEG6000 and lipid phase to 95 C;
b. Date syrup was heated to 75 C;
c. 10% of the date syrup was heated to 70 C with potassium sorbate,
solubilised, and
cooled to room temperature;
d. The date syrup was added to lipid phase and mixed with a spatula;
e. The mixture was homogenized with SiIverson L4RT emulsor screens;
f. The lipid phase and date syrup were homogenized at 6000 rpm for 1 mm;
g. The lipid phase and date syrup were further homogenized at 5000 rpm for 3
min;
h. The homogenization speed was reduced to 4000 rpm for 4 min with cooling in
a water
bath to 56 C;
i. Room temperature honey and potassium sorbate were added;
J. The mixture was mixed manually.
CUR66-3 was produced as follows (production of 1.5 kg):
a. Curcumin was melted with PEG6000 and lipid phase to 90 C;
b. Date syrup was heated to 70 C;
C. 10% of the date syrup was heated to 70 C with potassium sorbate,
solubilised and
cooled to room temperature;
d. The lipid phase was homogenized at 6000 rpm for about 20 s;
e. Date syrup was added slowly with homogenization for 3 min;
f. The lipid phase and date syrup were homogenized at 5000 rpm for 3 min;
g. The homogenization speed was reduced to 4000 rpm for 4 min with cooling in
a water
bath;
h. Honey and potassium sorbate were added at room temperature;
i. The mixture was mixed manually.
All three formulations produced a smooth, semisolid product with a sweet
taste.
In all three formulations microscopic observation revealed a few 10-25 pm
curcumin crystals
and lipid particles up to 3 pm.
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All the formulations were subjected to an accelerated stability test
consisting of storage at a
temperature of 32 C or 40 C for three months and monitoring of physical
stability by visual
inspection and of curcumin chemical stability by HPLC assay at zero time and
after three
months' storage. All formulations found stable at 32 C and some showed slight
creaming at
40 C. Curcumin was found to be stable with no detectable chemical
degradation.
Table 18: production with Pomegranate oil (CUR65), with Avocado oil (CUR66)
and with
Olive oil (CUR67)
Rational production Pilot production Pilot
production Pilot
CUR66-1 CUR66-2 CUR66-3
%W/W %W/W %W/W
Curcumin 4 4 4
Avocado oil 5 5 5
Polyethylene
Glycol 6000 6 6 6
Sucrose Ester
HLB 15 1 1
Polysorbate 80 0.6 0.5 0.5
Polyglycery1-3-
Dioleate 1 1 1
Potassium sorbate 0.05 0.05 0.05
Cold Honey 51 51 51
hot date syrup 31.35 31.45 31.45
SUM 100 100 100
Curcumin was melted
with PEG6000 to 90C. Curcumin was melted Curcumin was melted
Lipid phase added to with PEG6000 and with PEG6000 and lipid
Production: melted curcumin lipid phase to 95C phase
to 90C
Tests ZT:
Crystals some 10-25 micron some 10-25micron some 10-
25micron
submicron to 3micron, submicron to 3micron,
Emulsification submicron to 3micron very few 6micron very
few 6micron
State semi solid semi solid semi
solid
Smell good good good
Mixing with
water good good good
taste sweet sweet sweet
texture smooth smooth smooth
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CA 02808710 2013-02-18
WO 2012/023142 PCT/1L2011/000676
Curcumin HPLC assay Zero time
1: The quantitative results of Ctretunine:
Tam( % Curcumine
no. Sample name, IIPLCAN flteD
name (w/w)
IAI Room temperature 111975
LL1974/15,31 3.8
66(1) 17/3 production 2011-10996-2
IBI Room temperature 111976
2 1.1.1974A6,232 4.2
66(2) 17/3 production 2011-10996-3
ICI Room temperature LL 1977
3 LL1974/17,33 4.0
66(3) 17/3 production 2011-109964
Curcumin HPLC assay after three months storage at 40 C
1: The quantitative results of Curcurnine:
Tani % Curcu mine
Sample name HPLC/LTV ftles.D
name (wiw)
LL2128
1 CUR 66(1) LL2128q3-18 4.4
2011-11892-1
LL2129
2 66(2) 17/3 2011-11892-2 L1.2128/20-25 4.2
LL2130
3 66(3) 17/3 LL211.27-32 4.3
2011-11892-3
EXAMPLE 6
Curcumin is known to possess poor systemic availability, in his natural form,
i.e. mixed in
drinks or food, therefore an in vivo model was used to determine the extent to
which the
formulations of the present invention increase its oral bioavailability.
In the tests, male Wistar rats received 400 mg/kg of either unformulated
curcumin or
curcumin formulated in the solid lipid particles (SLP) formulation, according
to the present
invention, by oral gavage. Rats were killed at 15, 30, 60 and 120 min post
administration.
Plasma, intestinal mucosa and liver were analyzed for the presence of curcumin
using HPLC
with UV detection.
44
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WO 2012/023142 CA 02808710 2013-02-18PCT/1L2011/000676
Curcumin was identified in plasma, intestinal mucosa and liver of rats which
had received the
solid lipid particles (SLP) formulated curcumin. The curcumin plasma level
peak after
administration of the SLP formulated curcumin was tenfold higher than the
equivalent values
seen after unformulated curcumin. Similarly, liver levels of curcumin were
higher after
administration of SLP formulated curcumin as compared to unformulated
curcumin. In
contrast, curcumin concentrations in the gastrointestinal mucosa after
ingestion of the SLP
formulated curcumin were somewhat lower than those observed after
administration of
unformulated curcumin.
The results indicate that SLP formulated curcumin significantly increases
systemic
absorption of curcumin than the unformulated form. This formulation presents a
great
solution for compounds that posse's high medicinal benefit with poor systemic
availability.
This study emphasis the contribution of this specific formulation as a
delivery formulation, in
comparison to another product named "Mariva8".
The formulation of Mariva is basically Curcumin (CAS 458-37-7) and curcumin
phospholipid complex. The preparation of Meriva was performed byusing
EpiKuronTM 130
P, a de-oiled, powdered soybean lecithin enriched with 30%
phosphatidylcholine. Meriva
contained 16.89% curcuminoids, of which 93.82% was curcumin, the ratio of
curcumin to
Epikuronm4 130 P was 1:4.
"Meriva " is been marketed by a company whom conducted the same protocol of
clinical
study in purpose to published a new formulation for curcumin, whose added
value is to
increase the curcumin absorption through the mucosa tissue into the blood
stream (See
Timothy H. Marczylo et al., Cancer Chemother Pharmacol, 2007, 60:171-177).
When the two studies are being compared the results from the present invention
clinical study
with the SLP formulation showed higher curcumin concentration that was
absorbed into the
plasma and liver than the results of those of the "Meriva ". Furthermore the
incline in the
curcumin absorption that presented in the present invention is moderated and
continuing to
rise up to 60 min after oral administration while the 'Meriva " published
results present a
steep incline that stopped already 15 min after administration. This
comparison emphasis the
innovatively and the benefits of the SLP formulation, as provided by the
present invention,
that enable a continuing ongoing absorption which increase moderately up to 60
min after
oral administration, therefore provides stability to the active compound. The
stability quality
enable long active period for the compound in the tissues.
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WO 2012/023142 CA 02808710 2013-02-18PCT/1L2011/000676
To emphasis the improvement absorption of the SLP formulation versus the
Meriva product
we added two tables that present the delta of the curcumin concentration in
each time point.
Reference is now made to FIG. 5a, which shows plasma curcumin levels in rats
that had
received 400 mg/kg curcumin by oral gavage as a function of time following the
administration of the curcumin. The solid line shows results for rats that
received SLP
formulated curcumin, while the broken line shows results for results that
received
unformulated curcumin (represented as C3).
Reference is now made to FIG. 5b, which shows liver curcumin levels in rats
that had
received 400 mg/kg curcumin by oral gavage as a function of time following the
administration of the curcumin. The solid line shows results for rats that
received SLP
formulated curcumin, while the broken line shows results for results that
received
unformulated curcumin (represented as C3).
Reference is now made to FIG. Sc, which shows curcumin levels in the
gastrointestinal
mucosa of rats that had received 400 mg/kg curcumin by oral gavage as a
function of time
following the administration of the curcumin. The solid line shows results for
rats that
received SLP formulated curcumin, while the broken line shows results for
results that
received from unformulated curcumin (i.e., the control, represented as C3).
Reference is now made to FIG. 6a, which illustrates the delta in the plasma
curcumin
concentration while using the SLP and Meriva as a function of time (15
minutes, 30 minutes,
60 minutes and 120 minutes).
The SLP concentration was calculated was as follows: SLP formulation
concentration minus
C3 (i.e., the control) curcumin concentration.
The Meriva concentration was calculated was as follows: Meriva formulation
concentration
minus C3 (i.e., the control) curcumin concentration.
The delta was calculated for each time point. The units of the numbers are
ng/ml.
The minus mark appears where the C3 curcumin concentration is higher than the
SLP/
Meriva curcumin concentration.
Reference is now made to FIG. 6b, which illustrates the delta in the mucosa
curcumin
concentration while using the SLP and Meriva as a function of time (15
minutes, 30 minutes,
60 minutes and 120 minutes).
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The SLP concentration was calculated was as follows: SLP formulation
concentration minus
C3 (i.e., the control) curcumin concentration.
The Meriva concentration was calculated was as follows: Meriva formulation
concentration
minus C3 (i.e., the control) curcumin concentration.
The delta was calculated for each time point. The units of the numbers are
mg/g. The minus
mark appears where the C3 curcumin concentration is higher than the SLP/
Meriva curcumin
concentration.
Conclusions
Hot melt procedures for emulsification of curcumin-filled solid lipid
particles in honey were
developed and tested. Solid lipid particles produced by hot melt
emulsification were the
method of choice for curcumin incorporation in honey. The present inventors
have shown
that curcumin is soluble in mixtures of ethylene oxide polymers and lipids.
The present
inventors have also shown that free curcumin forms curcumin crystals in honey
and water.
Testing of several product parameters has shown that curcumin solubility is
pivotal to
product quality and that use of hydrogenated castor oil, steam, and palm oil
as lipid solvent
components is preferred since it results in a curcumin-in-honey formulation
that does not
have an aftertaste, exhibits smooth texture, has a honey smell and includes
substantially all of
the curcumin in the lipid microparticles suspended in the honey base.
It is appreciated that certain features of the invention, which are, for
clarity, described in the
context of separate embodiments, may also be provided in combination in a
single
embodiment. Conversely, various features of the invention, which are, for
brevity, described
in the context of a single embodiment, may also be provided separately or in
any suitable
subcombination.
Although the invention has been described in conjunction with specific
embodiments thereof,
it is evident that many alternatives, modifications and variations will be
apparent to those
skilled in the art. Accordingly, it is intended to embrace all such
alternatives, modifications
and variations that fall within the scope of the appended claims, with the
proper scope
determined only by the broadest interpretation of the claims.
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All publications, patents and patent applications mentioned in this
specification are herein
incorporated in their entirety by reference into the specification, to the
same extent as if each
individual publication, patent or patent application was specifically and
individually indicated
to be incorporated herein by reference. In addition, citation or
identification of any reference
in this application shall not be construed as an admission that such reference
is available as
prior art to the present invention.
48
