U.S. patent application number 14/725624 was filed with the patent office on 2015-09-17 for curcumin-er, a liposomal-plga sustained release nanocurcumin for minimizing qt prolongation for cancer therapy.
The applicant listed for this patent is SignPath Pharma Inc., University of North Texas Health Science Center. Invention is credited to Lawrence Helson, Anindita Mukerjee, Amalendu Prakash Ranjan, Jamboor K. Vishwanatha.
Application Number | 20150258026 14/725624 |
Document ID | / |
Family ID | 50184475 |
Filed Date | 2015-09-17 |
United States Patent
Application |
20150258026 |
Kind Code |
A1 |
Ranjan; Amalendu Prakash ;
et al. |
September 17, 2015 |
CURCUMIN-ER, A LIPOSOMAL-PLGA SUSTAINED RELEASE NANOCURCUMIN FOR
MINIMIZING QT PROLONGATION FOR CANCER THERAPY
Abstract
The present invention includes compositions and methods of
making a nanoparticle composition comprising a polymeric core
comprising one or more polymers and one or more active agents, and
at least one layer of one or more lipids on the surface of the
polymeric core; more specifically, the invention relates to the use
of curcumin within such a lipid-polymer nanoparticle formulation
for minimizing QT prolongation associated with curcumin in
treatment of cancer.
Inventors: |
Ranjan; Amalendu Prakash;
(Fort Worth, TX) ; Mukerjee; Anindita; (Fort
Worth, TX) ; Vishwanatha; Jamboor K.; (Fort Worth,
TX) ; Helson; Lawrence; (Quakertown, PA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of North Texas Health Science Center
SignPath Pharma Inc. |
Fort Worth
Quakertown |
TX
PA |
US
US |
|
|
Family ID: |
50184475 |
Appl. No.: |
14/725624 |
Filed: |
May 29, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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14016056 |
Aug 31, 2013 |
|
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14725624 |
|
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61695827 |
Aug 31, 2012 |
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Current U.S.
Class: |
424/1.11 ;
424/130.1; 424/450; 514/44A; 514/679 |
Current CPC
Class: |
A61K 47/34 20130101;
A61K 45/06 20130101; A61P 35/00 20180101; A61K 9/5123 20130101;
A61K 9/145 20130101; A61K 9/1277 20130101; A61K 9/5153 20130101;
A61K 9/1271 20130101; A61K 9/141 20130101; A61K 31/12 20130101 |
International
Class: |
A61K 9/127 20060101
A61K009/127; A61K 45/06 20060101 A61K045/06; A61K 47/34 20060101
A61K047/34; A61K 31/12 20060101 A61K031/12 |
Claims
1. A composition for treating cancer comprising: a polymeric
nanoparticle core comprising one or more polymers and at least one
of curcumin or curcuminoids; and at least one layer of one or more
lipids on the surface of the polymeric core, wherein the at least
one of the curcumin or curcuminoids nanoparticles, wherein the
composition does not cause QT prolongation when provided to a
subject.
2. The nanoparticle composition of claim 1, wherein the one or more
polymers comprise at least one of poly(lactic-co-glycolic acid)
(PLGA), poly(lactic acid), polylactide (PLA), or
poly-L-lactide-co-.epsilon.-caprolactone (PLCL).
3. The nanoparticle composition of claim 1, wherein the composition
further comprises an active agent selected from at least one of an
anti-cancer drug, an antibiotic, an antiviral, an antifungal, an
antihelminthic, a nutrient, a small molecule, a siRNA, an
antioxidant, and an antibody, or a conventional radioisotope.
4. The nanoparticle composition of claim 1, wherein the one or more
lipids comprise at least one of dimyristoyl phosphatidylcholine
(DMPC), dimyristoyl phosphatidylglycerol (DMPG),
1,2-dioctadecanoyl-sn-glycero-3-phosphoethanolamine (DSPE),
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene
glycol) (DSPE-PEG), DMPE PEG, maleimide, lecithin, cholesterol,
1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine
rhodamine B sulfonyl) (ammonium salt), and
1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxa-
diazol-4-yl) (ammonium salt).
5. The nanoparticle composition of claim 1, wherein the one or more
lipids comprise dimyristoyl phosphatidylcholine (DMPC) and
dimyristoyl phosphatidylglycerol (DMPG) in a molar ratio of 9:1,
7:3, 8:2, or 7.5:2.5.
6. The nanoparticle composition of claim 1, further comprising at
least one targeting agent, wherein the targeting agent selectively
targets the nanoparticle to diseased tissue/cells, thereby
minimizing whole body dose.
7. The nanoparticle composition of claim 1, further comprising at
least one targeting agent, wherein the targeting agent comprises an
antibody or functional fragment thereof, a small molecule, a
peptide, a carbohydrate, an siRNA, a protein, a nucleic acid, an
aptamer, a second nanoparticle, a cytokine, a chemokine, a
lymphokine, a receptor, a lipid, a lectin, a ferrous metal, a
magnetic particle, a linker, an isotope and combinations
thereof.
8. The nanoparticle composition of claim 1, wherein the
nanoparticles have a size of 90 to 150 nm.
9. The nanoparticle composition of claim 1, wherein a
bioavailability of the active agent is increased, a QT prolongation
is reduced, and the active agent is released in a sustained
manner.
10. The nanoparticle composition of claim 1, wherein the
nanoparticles are adapted for intramuscular, subcutaneous,
intravascular, or intravenous administration.
11. A method of forming a nanoparticle composition comprising:
forming an organic phase by combining one or more polymers, one or
more solvents and at least one of curcumin or curcuminoids; forming
a lipid aqueous phase by mixing one or more lipids with water;
mixing the organic phase with the lipid aqueous phase, whereby an
emulsion is formed; and incubating the emulsion, whereby
self-assembly of nanoparticles occurs and wherein the curcumin or
curcuminoids nanoparticles does not cause QT prolongation when
provided to a subject.
12. A method for treating a patient suspected of being afflicted
with a disease comprising administering nanoparticles, wherein the
nanoparticles comprise a polymeric core comprising one or more
polymers and one or more active agents and at least one layer of
one or more lipids on the surface of the polymeric core, wherein
the active agent is suspected of causing QT prolongation when
provided to a subject.
13. The method of claim 12, wherein administering nanoparticles
comprises administering the nanoparticle by intramuscular,
subcutaneous, intravascular, or intravenous administration.
14. The method of claim 12, wherein the disease is selected from
the group consisting of neurologic, oncologic, and metabolic
diseases.
15. The method of claim 12, wherein the disease is selected from
the group consisting of Parkinson's disease, Alzheimer's disease,
multiple sclerosis, ALS, sequel, behavioral and cognitive
disorders, autism spectrum, depression, and neoplastic disease.
16. The method of claim 12, wherein the active agent is released in
a sustained manner.
17. A pharmaceutical agent comprising: a nanoparticle for drug
delivery comprising a polymer, an active agent that causes QT
prolongation, and at least one layer of one or more lipids
encapsulating the polymer and the active agent and the agent does
not cause QT prolongation.
18. A method for treating a patient suspected of being afflicted
with a disease, the method comprising administering nanoparticles,
wherein the nanoparticles comprise a polymeric core comprising one
or more polymers, curcumin, and at least one layer of one or more
lipids on the surface of the polymeric core, wherein treating the
patient does not cause QT prolongation.
19. A method of treating a subject suspected of having cancer
comprising: identifying that a patient suspected of having a
cancer; and Providing the subject with an amount of at least one or
curcumin or curcuminoids in an amount sufficient to reduce the
cancer in the subject, wherein the at least one or curcumin or
curcuminoids are in a polymeric nanoparticle core comprising one or
more polymers and at least one of curcumin or curcuminoids; and at
least one layer of one or more lipids on the surface of the
polymeric core, wherein the at least one of the curcumin or
curcuminoids nanoparticles does not cause QT prolongation when
provided to a subject.
20. The method of claim 19, wherein the cancer is a pancreatic, a
prostate, or a breast cancer.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of Ser. No.
14/016,056 filed Aug. 31, 2013, which claims priority to U.S.
Provisional Patent Application Ser. No. 61/695,827 filed Aug. 31,
2012, the entire contents of which are incorporated herein by
reference.
TECHNICAL FIELD OF THE INVENTION
[0002] The present invention relates in general to nanoparticles
comprising a polymeric core comprising one or more polymers and one
or more active agents and at least one layer of one or more lipids
on the surface of the polymeric core. More specifically, the
invention relates to the use of curcumin within such a
lipid-polymer nanoparticle formulation for minimizing QT
prolongation associated with curcumin in treatment of cancer.
STATEMENT OF FEDERALLY FUNDED RESEARCH
[0003] None.
BACKGROUND OF THE INVENTION
[0004] Without limiting the scope of the invention, its background
is described in connection with the delivery of active
pharmaceutical agents.
[0005] U.S. Pat. No. 7,968,115 to Kurzrock (filed Sep. 7, 2005) is
said to provide a compositions and methods for the treatment of
cancer, including pancreatic cancer, breast cancer and melanoma, in
a human patient. The methods and compositions of the present
invention employ curcumin or a curcumin analogue encapsulated in a
colloidal drug delivery system, preferably a liposomal drug
delivery system. Suitable colloidal drug delivery systems also
include nanoparticles, nanocapsules, microparticles or block
copolymer micelles. The colloidal drug delivery system
encapsulating curcumin or a curcumin analogue is administered
parenterally in a pharmaceutically acceptable carrier.
[0006] U.S. Pat. No. 8,202,839 to Sung (filed Jan. 7, 2012) is said
to disclose a pharmaceutical composition of bioactive nanoparticles
composed of chitosan, poly-glutamic acid, and a bioactive agent for
oral delivery. The chitosan-based nanoparticles are characterized
with a positive surface charge and enhanced permeability for oral
drug delivery.
[0007] U.S. Patent Application Publication Number 20120058208 by
Jacob (Synergistic Composition for Enhancing Bioavailability of
Curcumin) (filed Mar. 8, 2012) is said to relate to a composition
to enhance the bioavailability of curcumin. In one embodiment, a
composition comprising plant extracts of curcumin, vanilla and
ginger, wherein the extracts of ginger and vanilla are rich in
gingerol and vanillin respectively, is provided. In other
embodiments, curcumin, and one or more items selected from the
group of vanilla, ginger and capsaicin is provided.
[0008] U.S. Patent Application Publication Number 20120003177 by
Shen (Curcumin-containing polymers and water-soluble curcumin
derivatives as prodrugs of prodrug carriers) (filed Jan. 5, 2012)
is said to describe Curcumin, a polyphenol extracted from the
rhizome turmeric, that has been polymerized to produce a polymer
material having a backbone of one or more repeating structural
units, at least one of which comprises a curcumin monomer residue.
These curcumin-containing polymers have a wide range of
pharmacological activities, including, among others antitumor,
antioxidant, anti-inflammatory, antithrombotic and antibacterial
activities. Certain species of these polymers have exhibited
remarkable antitumor activity. Water-soluble curcumin derivatives
and their use as prodrugs and prodrug carriers are also
disclosed.
SUMMARY OF THE INVENTION
[0009] Problems associated with Curcumin are low solubility, low
bioavailability, QT prolongation, and fast in vivo clearance. The
advantages of liposomal nanocurcumin are no QT prolongation, high
bioavailability, and low in vivo clearance, but the disadvantages
are rapid release. The advantages of polymeric nanocurcumin are
high bioavailability, sustained release, and low in vivo clearance,
but the disadvantages are QT prolongation. The advantages of hybrid
nanocurcumin are high bioavailability, sustained release, no QT
prolongation, and low in vivo clearance.
[0010] The present invention includes methods and compositions
comprising a polymeric nanoparticle core comprising one or more
polymers and one or more active agents; and at least one layer of
one or more lipids on the surface of the polymeric core. The one or
more polymers may comprise PLGA; and/or at least one polymer
selected from the group consisting of poly(lactic acid),
polylactide (PLA), and poly-L-lactide-co-.epsilon.-caprolactone
(PLCL). In certain aspects, the one or more active agents comprise
curcumin or a curcuminoid. The active agent may comprise at least
one anti-cancer drug; and/or be selected from at least one of an
anti-cancer drug, an antibiotic, an antiviral, an antifungal, an
antihelminthic, a nutrient, a small molecule, a siRNA, an
antioxidant, and an antibody. In certain aspects, the nanoparticle
composition does not cause QT prolongation. In certain aspects, the
nanoparticle composition has high bioavailability. In certain
aspects, the active agent may comprise a conventional radioisotope.
The one or more active agents comprise a water-insoluble dye;
and/or a metal nanoparticle, to be used as contrast agents for MRI;
and/or be selected from the group comprising Nile red, iron, and
platinum. In certain aspects, the one or more lipids comprise
1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC); and/or
dimyristoyl phosphatidylglycerol (DMPG);
1,2-dioctadecanoyl-sn-glycero-3-phosphoethanolamine (DSPE),
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene
glycol) (DSPE-PEG), DMPE PEG Maleimide, Lecithin, cholesterol,
1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine
rhodamine B sulfonyl) (ammonium salt), and
1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxa-
diazol-4-yl) (ammonium salt). In various aspects, the nanoparticle
composition may comprise DMPC and DMPG in a molar ratio of 9:1,
7:3, 8:2, or 7.5:2.5. In certain aspects, the nanoparticles may
comprise at least one targeting agent, wherein the targeting agent
selectively targets the nanoparticle to diseased tissue/cells,
thereby minimizing whole body dose; and/or wherein the targeting
agent comprises an antibody or functional fragment thereof that is
capable of recognizing a target antigen; and/or selected from the
group consisting of an antibody, a small molecule, a peptide, a
carbohydrate, an siRNA, a protein, a nucleic acid, an aptamer, a
second nanoparticle, a cytokine, a chemokine, a lymphokine, a
receptor, a lipid, a lectin, a ferrous metal, a magnetic particle,
a linker, an isotope and combinations thereof. In certain aspects,
the nanoparticles have a size of 90 to 150 nm. The bioavailability
of the active agent may be increased, a QT prolongation is reduced,
and the active agent may be released in a sustained manner.
[0011] The invention includes embodiments of methods for forming a
nanoparticle composition comprising forming an organic phase by
combining one or more polymers, one or more solvents and one or
more active agents; forming a lipid aqueous phase by mixing one or
more lipids with water; mixing the organic phase with the lipid
aqueous phase, whereby an emulsion is formed; and incubating the
emulsion, whereby self-assembly of nanoparticles occurs. The one or
more polymers may comprise PLGA; and/or at least one polymer
selected from the group consisting of poly(lactic acid),
polylactide (PLA), and poly-L-lactide-co-.epsilon.-caprolactone
(PLCL). The organic phase may comprise PLGA in a concentration of
2-90 mg/ml; and/or curcumin in a concentration of 1-15
weight/volume %. In various aspects, the one or more solvents may
comprise an organic solvent; acetonitrile; at least one solvent
selected from the group consisting of Acetone, tert butyl alcohol,
Dimethyl formamide, and Hexafluro isopropanol. The one or more
active agents comprise curcumin or a curcuminoid; and/or at least
one anti-cancer drug; and/or a conventional radioisotope; and/or at
least one active agent selected from the group consisting of
selected from the group comprising Nile red, iron, and platinum. In
certain aspects, the one or more lipids may comprise DMPC; and/or
DMPG, and/or at least one lipid selected from the group consisting
of 1,2-dioctadecanoyl-sn-glycero-3-phosphoethanolamine (DSPE),
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethy-
lene glycol) (DSPE-PEG), DMPE PEG Maleimide, Lecithin, cholesterol,
1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine
rhodamine B sulfonyl) (ammonium salt), and
1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxa-
diazol-4-yl) (ammonium salt). In certain aspects, the one or more
lipids comprise DMPC and DMPG in a molar ratio of 9:1, 7:3, 8:2,
7.5:2.5. In certain aspects, mixing the organic phase with the
lipid aqueous phase comprises slowly stirring the organic phase
into the lipid aqueous phase; and/or mixing the organic phase with
the lipid aqueous phase comprises vortexing; and/or mixing the
organic phase with the lipid aqueous phase further comprises
sonicating. In certain aspects, incubating the emulsion comprises
stirring the emulsion for 2 hours. In certain aspects, the method
may further comprise separating the nanoparticles after incubating
the emulsion; and/or filtering the nanoparticles after incubating
the emulsion; and/or freezing the nanoparticles; and/or
lyophilizing the nanoparticles; and/or attaching a targeting agent
to the nanoparticles; and/or attaching at least one targeting
agent, wherein the targeting agent selectively targets the
nanoparticle to diseased tissue/cells, thereby minimizing whole
body dose; and/or attaching at least one targeting agent to the
nanoparticles, wherein the targeting agent comprises an antibody or
functional fragment thereof that is capable of recognizing a target
antigen. In certain aspects, the nanoparticles have a size of 90 to
150 nm.
[0012] The invention includes embodiments of pharmaceutical agents
comprising a nanoparticle for drug delivery comprising a polymer,
an active agent and at least one layer of one or more lipids
encapsulating the polymer and the active agent.
[0013] The invention includes embodiments for treating a patient
suspected of being afflicted with a disease comprising
administering nanoparticles, wherein the nanoparticles comprise a
polymeric core comprising one or more polymers and one or more
active agents and at least one layer of one or more lipids on the
surface of the polymeric core. In certain aspects, administering
nanoparticles comprises administering the nanoparticle by
intramuscular, subcutaneous, intravascular, or intravenous
administration. Disease can be selected from the group consisting
of neurologic, oncologic, and metabolic diseases; and/or from the
group consisting of Parkinson's disease, Alzheimer's disease,
multiple sclerosis, ALS, sequel, behavioral and cognitive
disorders, autism spectrum, depression, and neoplastic disease;
and/or cancer. In certain aspects, the active agent is released in
a sustained manner.
[0014] The invention includes embodiments of composition comprising
a polymeric nanoparticle core comprising one or more polymers and
curcumin and at least one layer of one or more lipids on the
surface of the polymeric core.
[0015] The invention includes embodiments of forming a nanoparticle
composition comprising forming an organic phase by combining one or
more polymers, one or more solvents and curcumin; forming a lipid
aqueous phase by mixing one or more lipids with water; mixing the
organic phase with the lipid aqueous phase, whereby an emulsion is
formed; and incubating the emulsion, whereby self-assembly of
nanoparticles occurs.
[0016] The invention includes embodiments of pharmaceutical agents
comprising a nanoparticle for drug delivery comprising a polymer,
curcumin, and at least one layer of one or more lipids
encapsulating the polymer and the active agent.
[0017] The invention includes embodiments of methods for treating a
patient suspected of being afflicted with a disease, the method
comprising administering nanoparticles, wherein the nanoparticles
comprise a polymeric core comprising one or more polymers,
curcumin, and at least one layer of one or more lipids on the
surface of the polymeric core.
[0018] Another embodiment includes a composition for treating
cancer comprising: a polymeric nanoparticle core comprising one or
more polymers and at least one of curcumin or curcuminoids; and at
least one layer of one or more lipids on the surface of the
polymeric core, wherein the at least one of the curcumin or
curcuminoids nanoparticles, wherein the composition does not cause
QT prolongation when provided to a subject. In one aspect, the one
or more polymers comprise at least one of poly(lactic-co-glycolic
acid) (PLGA), poly(lactic acid), polylactide (PLA), or
poly-L-lactide-co-.epsilon.-caprolactone (PLCL).
[0019] Another embodiment includes a method of forming a
nanoparticle composition comprising: forming an organic phase by
combining one or more polymers, one or more solvents and at least
one of curcumin or curcuminoids; forming a lipid aqueous phase by
mixing one or more lipids with water; mixing the organic phase with
the lipid aqueous phase, whereby an emulsion is formed; and
incubating the emulsion, whereby self-assembly of nanoparticles
occurs and wherein the curcumin or curcuminoids nanoparticles does
not cause QT prolongation when provided to a subject.
[0020] Another embodiment includes a method for treating a patient
suspected of being afflicted with a disease comprising
administering nanoparticles, wherein the nanoparticles comprise a
polymeric core comprising one or more polymers and one or more
active agents and at least one layer of one or more lipids on the
surface of the polymeric core, wherein the active agent is
suspected of causing QT prolongation when provided to a subject. In
one aspect, the method also includes the step of administering the
nanoparticle by intramuscular, subcutaneous, intravascular, or
intravenous administration.
[0021] Another embodiment includes a method of forming a
nanoparticle that prevents the active agent from causing QT
prolongation composition comprising: forming an organic phase by
combining one or more polymers, one or more solvents and the active
agent that causes QT prolongation; forming a lipid aqueous phase by
mixing one or more lipids with water; mixing the organic phase with
the lipid aqueous phase, whereby an emulsion is formed; and
incubating the emulsion, whereby self-assembly of nanoparticles
occurs.
[0022] Another embodiment includes a pharmaceutical agent
comprising: a nanoparticle for drug delivery comprising a polymer,
an active agent that causes QT prolongation, and at least one layer
of one or more lipids encapsulating the polymer and the active
agent and the agent does not cause QT prolongation.
[0023] Another embodiment includes a method for treating a patient
suspected of being afflicted with a disease, the method comprising
administering nanoparticles, wherein the nanoparticles comprise a
polymeric core comprising one or more polymers, curcumin, and at
least one layer of one or more lipids on the surface of the
polymeric core, wherein treating the patient does not cause QT
prolongation.
[0024] In another embodiment, the method of treating a subject
suspected of having cancer includes: identifying that a patient
suspected of having a cancer; and Providing the subject with an
amount of at least one or curcumin or curcuminoids in an amount
sufficient to reduce the cancer in the subject, wherein the at
least one or curcumin or curcuminoids are in a polymeric
nanoparticle core comprising one or more polymers and at least one
of curcumin or curcuminoids; and at least one layer of one or more
lipids on the surface of the polymeric core, wherein the at least
one of the curcumin or curcuminoids nanoparticles does not cause QT
prolongation when provided to a subject. In one aspect, the cancer
is a pancreatic, prostate or breast cancer.
BRIEF DESCRIPTION OF THE DRAWINGS
[0025] For a more complete understanding of the features and
advantages of the present invention, reference is now made to the
detailed description of the invention along with the accompanying
figures and in which:
[0026] FIG. 1 depicts the basic concept of hybrid nanocurcumin
(HNC) formation; Lipids-DMPC and DMPG. Problems associated with
Curcumin are low solubility, low bioavailability, QT prolongation,
and fast in vivo clearance. The advantages of liposomal
nanocurcumin are no QT prolongation, high bioavailability, and low
in vivo clearance, but the disadvantages are rapid release. The
advantages of polymeric nanocurcumin are high bioavailability,
sustained release, and low in vivo clearance, but the disadvantages
are QT prolongation. The advantages of hybrid nanocurcumin are high
bioavailability, sustained release, no QT prolongation, and low in
vivo clearance.
[0027] FIG. 2 demonstrates improved dispersibility in water with
HNC.
[0028] FIG. 3 represents transmission electron micrographs showing
HNC. The TEM scan shows HNC as spherical smooth nanoparticles with
uniform size.
[0029] FIGS. 4A and 4B: FIG. 4A shows formulations of hybrid
nanocurcumin (HNC). Demonstrated are four different formulations of
HNC using different ratio of DMPC and DMPG. FIG. 4B shows particle
size distribution of Batch 3.
[0030] FIG. 5 shows HNC characterization, including average
particle size, drug loading, and encapsulation efficiency.
[0031] FIG. 6 shows hERG current density analysis of curcumin;
liposomal curcumin; and PLGA curcumin.
[0032] FIG. 7 shows hERG current density analysis of
liposomes+curcumin; and liposomes.
[0033] FIG. 8 shows intracellular uptake of HNC in MiaPaCa
cells.
[0034] FIG. 9 shows Western blot analysis of MiaPaCa cells treated
with hybrid nanocurcumin (25 .mu.M (micromolar)). Lane 1:
Untreated; lane 2: Blank nanoparticle; lane 3: Curcumin (24 hrs);
lane 4: HNC (24 hrs) and; lane 5 HNC (48 hrs).
[0035] FIG. 10 shows MTT cell viability using HNC employing a
pancreatic cancer cell line (MiaPaCa cell line) at 48 hours.
[0036] FIG. 11 shows the pulses protocol or the original data
acquisition design: Acquisition Rate(s): 1.0 kHz
[0037] FIG. 12 shows the effect of batch A on hERG current density
from transfected HEK 293 cells at 20 mV.
[0038] FIG. 13 shows the effect of batch A on hERG current density
from transfected HEK 293 cells at 20 mV.
[0039] FIG. 14 shows the relationship (I-V) of hERG current
amplitude from transfected HEK 293 cells exposed to Batch A.
[0040] FIG. 15 shows the effect of batch B on hERG current density
from transfected HEK 293 cells at 20 mV.
[0041] FIG. 16 shows the effect of batch B on hERG current density
from transfected HEK 293 cells at 20 mV.
[0042] FIG. 17 shows the relationship (I-V) of hERG current
amplitude from transfected HEK 293 cells exposed to Batch B.
[0043] FIG. 18 shows the effect of batch C on hERG current density
from transfected HEK 293 cells at 20 mV.
[0044] FIG. 19 shows the effect of Batch C on hERG current density
from transfected HEK 293 cells at 20 mV.
[0045] FIG. 20 shows the relationship (I-V) of hERG current
amplitude from transfected HEK 293 cells exposed to Batch C.
[0046] FIG. 21 shows the effect of batch D on hERG current density
from transfected HEK 293 cells at 20 mV.
[0047] FIG. 22 shows the effect of batch D on hERG current density
from transfected HEK 293 cells at 20 mV.
[0048] FIG. 23 shows the relationship (I-V) of hERG current
amplitude from transfected HEK 293 cells exposed to Batch D.
[0049] FIG. 24 shows effect of batch E on hERG current density from
transfected HEK 293 cells at 20 mV.
[0050] FIG. 25 shows the effect of batch E on hERG current density
from transfected HEK 293 cells at 20 mV.
[0051] FIG. 26 shows relationship (I-V) of hERG current amplitude
from transfected HEK 293 cells exposed to Batch E.
[0052] FIG. 27 shows the effect of tested compounds on hERG current
density at +20 mV.
[0053] FIG. 28 shows the results of the treatment of breast cancer
in a cancer xenograft mouse model system.
[0054] FIG. 29 shows additional results of the treatment of a
different breast cancer in a cancer xenograft mouse model
system.
DETAILED DESCRIPTION OF THE INVENTION
[0055] While the making and using of various embodiments of the
present invention are discussed in detail below, it should be
appreciated that the present invention provides many applicable
inventive concepts that can be embodied in a wide variety of
specific contexts. The specific embodiments discussed herein are
merely illustrative of specific ways to make and use the invention
and do not delimit the scope of the invention.
[0056] To facilitate the understanding of this invention, a number
of terms are defined below. Terms defined herein have meanings as
commonly understood by a person of ordinary skill in the areas
relevant to the present invention. Terms such as "a", "an" and
"the" are not intended to refer to only a singular entity, but
include the general class of which a specific example may be used
for illustration. The terminology herein is used to describe
specific embodiments of the invention, but their usage does not
delimit the invention, except as outlined in the claims.
[0057] Problems associated with Curcumin are low solubility, low
bioavailability, QT prolongation, and fast in vivo clearance. The
advantages of liposomal nanocurcumin are no QT prolongation, high
bioavailability, and low in vivo clearance, but the disadvantages
are rapid release. The advantages of polymeric nanocurcumin are
high bioavailability, sustained release, and low in vivo clearance,
but the disadvantages are QT prolongation. The advantages of hybrid
nanocurcumin are high bioavailability, sustained release, no QT
prolongation, and low in vivo clearance.
[0058] A requirement in commercial drug development is to assay
drug effects on hERG (Ikr) in in vitro assays using transfected
KEK293 cells. The present inventors determined anti-hERG activity
of curcumin (diferuloylmethane) in DMSO, and of three formulated
curcumin compounds: liposomal curcumin, nanocurcumin, and a
sustained release PLGA curcumin. The present inventors recognize
that the K+ current IC.sub.50 of curcumin formulated in DMSO is 3.4
uM. Considered within the context of current clinical Phase 1a
pharmacokinetics in normal subjects where blood plasma levels range
between 5-11 uMol, following a two hour infusion of 4.5 mg/kg,
intravenous, or subcutaneous curcumin formulations for therapeutic
applications can inhibit IKr, lead to Torsade de Points, and
possible clinical mortality. However, neither the liposomal, nor
the nanocurcumin formulation at 12 uMol exhibits this effect on the
K+ channel. The co-administration of empty liposomes to curcumin
was equally effective in prohibiting the hERG blockade, however,
the PLGA-curcumin formulation lacked this effect.
[0059] These observations are one basis for (constructing) a new
curcumin formulation consisting of liposome and PLGA, which allows
sustained release of curcumin without the associated cardiac K+
channel inhibitory properties of curcumin.
[0060] The treatment of cancer is limited by the side effects of
the anti-cancer drugs. Chemotherapy is the only available option
for the treatment of advanced cancers. However, increasing
evidences of drug resistance and non-specific toxicity of these
agents limits their therapeutic outcomes. To overcome this problem
it is important to deliver the drug at the site of cancer in the
body in the right amount. A novel way to approach this problem is
through targeted drug delivery system, which preferentially
delivers the drug to the site of cancer. In certain embodiment,
targeting molecules (e.g., antibodies) that recognize the cancer
cells and direct the drug containing tiny spherical particles
(nanoparticles) to the cancer cells are used.
[0061] In certain embodiments, at least one targeting agent is
attached to the nanoparticles, wherein the targeting agent
comprises an antibody or functional fragment thereof that is
capable of recognizing a target antigen. The targeting agents may
be attached by insertion of hetero/homo bifunctional spacer capable
of reacting with amines of lipids and targeting moieties.
[0062] Curcumin is a potent anticancer agent and is being used for
its pharmacological action for last few decades. However, the major
problems associated with curcumin are (1) low systemic
bioavailability following administration via any route; (2)
curcumin alone brings about QT prolongation; and (3) fast in vivo
clearance of curcumin. The present inventors solved these problems
by formulating curcumin (99% pure) into a hybrid nanoformulation.
See FIG. 1.
[0063] The present inventors recognized that a nanoformulation
provides the solutions to increase bioavailability and that
liposome formulation of curcumin show almost no QT prolongation.
But such formulations lack stability and possess some inherent
toxicity at higher doses. The present inventors recognize that
curcumin has a very rapid clearance when administered in animal
models.
[0064] The present inventors have developed a nanoformulation
system that increases the bioavailability of curcumin, minimizes
the QT prolongation, and releases the drug curcumin in a sustained
manner.
[0065] The hybrid nanocurcumin (HNC) system is a hybrid of lipids
and polymer wherein the polymeric core encapsulates curcumin. The
lipid is present as a continuous layer on the surface of the
polymeric nanoparticle. In other word, the lipid cases the
polymeric nanoparticle. The lipid component of the hybrid
nanocurcumin helps in reducing the QT prolongation while the
polymeric core of the hybrid system facilitates the release of
curcumin in a sustained manner. The hybrid nanocurcumin (HNC)
system solved all the above-mentioned problems of (1)
bioavailability of curcumin, (2) QT prolongation due to curcumin
and (3) sustained release of curcumin simultaneously.
[0066] The advantages of hybrid nanocurcumin (HNC) system are: (1)
in vivo bioavailability of active agents (e.g., curcumin) is
improved; (2) the lipid component of the hybrid nanocurcumin
reduces QT prolongation; (3) the polymeric core of the hybrid
system facilitates the release of the active agent (e.g., curcumin)
in a sustained manner; (4) the formulation itself is simple,
convenient one-step process; and (5) this system can be used to
formulate other similar type of drugs or active agents, which may
comprise hydrophobic molecules. Examples would include curcumin
analogues, docetaxel, paclitaxel etc.
[0067] The commercial potentials of hybrid nanocurcumin formulation
are enormous due to better bioavailability and reduced side
effects.
[0068] An embodiment is a Liposomal-Curcumin-PLGA sustained release
compound for prevention and treatment of neurologic, oncologic, or
metabolic diseases (Hybrid Nanocurcumin formulation).
[0069] Certain embodiments can be described as intravenous and/or
subcutaneous administration of a novel formulation of synthesized
curcumin (diferuloylmethane) bound to PLGA and a liposome. Such
formulation is designed to offer a sustained release of curcumin as
active agent. Reference is made to the prevention of cardiac events
due to the incorporation of a liposomal component of the
formulation.
[0070] In further embodiments the compositions may be used for the
treatment of neurologic-auto-immunological degenerative diseases
(Parkinson's disease, Alzheimer's disease, multiple sclerosis, ALS,
sequel, behavioral and cognitive disorders, autism spectrum, and
depression), neoplastic diseases (cancer).
[0071] In certain embodiments the compositions of the present
invention are administered intramuscular, subcutaneous and or
intravascular.
[0072] Certain embodiments comprise curcumin
(diferuloylmethane)-encapsulated in a liposomal--PLGA envelope
designated hybrid nanocurcumin formulation.
[0073] In one embodiment, the active agent is curcumin, which is a
potent natural anticancer agent, is employed in a
nanoparticle-based delivery system. One limitation is the QT
prolongation effect of curcumin, even when it is associated with
nanoparticle-based systems. This makes it difficult to pass FDA
standards for commercial use. The hybrid nanocurcumin formulation
solves this problem and reduces QT prolongation effect of curcumin,
which makes it ideal for commercial application. In addition, the
hybrid nanocurcumin formulation releases curcumin in a sustained
manner, which improves the systemic availability and decreases fast
clearance of curcumin in animal models. Therefore, the hybrid
nanocurcumin formulation can directly be used to produce
nanotechnology based hybrid dosage forms for curcumin. In other
embodiments curcumin may be replaced by a variety of similar drugs
or active agents. Such compositions may directly go into production
by pharmaceutical companies to test for phase I and phase II.
Example 1
[0074] Hybrid Nanocurcumin Formulation: PLGA was dissolved in
organic solvent, acetonitrile to get a concentration of 10 mg/ml.
Curcumin (5%) was dissolved in this polymer-organic solvent phase.
Lipids (DMPC and DMPG) were mixed in a different molar ratios and
volume was made up to 1 ml. In more detail:
[0075] Hybrid Nanocurcumin Formulation: Polymer PLGA (10 mg) was
dissolved in 1 ml of organic solvent, acetonitrile to get a
concentration of 10 mg/ml. Curcumin (5% with respect to polymer)
was dissolved in this polymer-organic solvent mixture. Lipids (DMPC
and DMPG) were mixed in different molar ratios, and it was found
that a ratio DMPC/DMPG=7.5/2.5 gave the best particles. DMPC (lipid
1) was dissolved in 4% ethanol in water. DMPG (lipid 2) was
dissolved in water and volume was made up to 1 ml. These solutions
were mixed and heated to obtained transparent solutions. Total
lipid content with respect to polymer was varied from 2 mg to 8 mg.
The organic phase was slowly stirred into the lipid aqueous phase
keeping the organic to aqueous volume ratio at 1:1. The emulsion
was vortexed for 30 sec and then sonicated for 5 min. The whole
emulsion system was then stirred for 2-3 hours for self-assembly.
This was then filtered thrice using Amicon filter (10 KD cutoff).
The hybrid particles thus obtained were flash frozen using liquid
nitrogen and lyophilized overnight. These were stored at
-20.degree. C. until further used.
[0076] The organic phase was slowly stirred into the lipid aqueous
phase keeping the organic to aqueous volume ratio at 1:1. The
emulsion was vortexed for 30 sec and then sonicated for 5 min. The
whole emulsion system was then stirred for 2 hours for
self-assembly. This was then filtered thrice using Amicon filter
(10 KD cutoff). The hybrid particles thus obtained were flash
frozen using liquid nitrogen and lyophilized overnight. These were
stored at -20.degree. C. until further used.
[0077] Hybrid Nanocurcumin Characterization: The hybrid
nanoparticles were characterized for particle size, drug loading,
encapsulation efficiency and surface morphology. FIG. 4A shows
results from one set of studies where the total amounts of lipids
were varied keeping the molar ratio of two lipids constant. In
other studies, lipids (DMPC and DMPG) were mixed in different molar
ratios and we found that DMPC/DMPG::7.5/2.5 gave the best
particles. In certain embodiments, the Hybrid Nanocurcumin is
referred to herein as Curcumin ER.
[0078] Particle size distribution: The particle size distribution
is shown in FIG. 4B. The particle sizes for various batches post
lyophilization are listed in Table 1. Particle size analysis of the
lyophilized nanoparticles was carried out using a Nanotrac system
(Mircotrac, Inc., Montgomeryville, Pa., USA). The lyophilized
nanoparticles were dispersed in double distilled water and vortexed
at high for 10 sec and then measured for particle size. The results
were reported as the average of three runs with triplicate runs in
each run.
TABLE-US-00001 TABLE 1 Average particle size distributions for all
batches Batch DMPC + DMPG (mg) Av. Particle Size (nm) Batch 1 2
138.0 Batch 2 4 117.2 Batch 3 6 142.7 Batch 4 8 103.6
[0079] Drug loading and encapsulation efficiency: The hybrid
nanocurcumin was dissolved in acetonitrile and drug loading and
encapsulation efficiency was determined by spectrophotometry.
Values are listed in Table 2. Lyophilized hybrid nanoparticles (5
mg) was dissolved in 2 ml acetonitrile to extract curcumin into
acetonitrile for determining the encapsulation efficiency. The
samples in acetonitrile were gently shaken on a shaker for 4 h at
room temperature to completely extract out curcumin from the
nanoparticles into acetonitrile. These solutions were centrifuged
at 14,000 rpm (Centrifuge 5415D, Eppendorf AG, Hamburg, Germany)
and supernatant was collected. Suspension (20 .mu.l) was dissolved
in ethanol (1 ml) and was used for the estimations. The curcumin
concentrations were measured spectrophotometrically at 450 nm. A
standard plot of curcumin (0-10 .mu.g/ml) was prepared under
identical conditions.
[0080] The encapsulation efficiency (EE) of PLGA-CURC was
calculated using
Encapsulation efficiency ( % ) = Total drug content in
nanoparticles Total drug amount .times. 100 ##EQU00001##
[0081] The percent drug loading was calculated by total amount of
drug extracted from the hybrid nanoparticles to the known weight of
nanoparticles
Drug loading ( % ) = Drug content Weight of nanoparticles .times.
100 ##EQU00002##
TABLE-US-00002 TABLE 2 Drug loading and encapsulation efficiency
for all batches. Batch Drug Loading (%) Encapsulation efficiency
(%) Batch 1 0.5 10 Batch 2 0.6 12 Batch 3 1.0 20 Batch 4 0.3 6
[0082] Surface morphology: Surface morphology of the HNC was
determined by Transmission electron microscopy. The TEM scan is
shown below in FIG. 3. The surface morphology of the hybrid
nanoparticle was studied using transmission electron microscopy,
(TEM). A small quantity of aqueous solution of the lyophilized
hybrid nanoparticles (1 mg/ml) was placed on a TEM grid surface
with a filter paper (Whatman No. 1). One drop of 1% uranyl acetate
was added to the surface of the carbon-coated grid. After 1 minute
of incubation, excess fluid was removed and the grid surface was
air dried at room temperature. It was then loaded into the
transmission electron microscope (LEO EM910, Carl Zeiss SMT Inc,
NY, USA) attached to a Gatan SC 1000 CCD camera. HNC are
characterized, which included determination of average particle
size, drug loading, and encapsulation efficiency and results are
shown in FIG. 5.
[0083] Hybrid Nanocurcumin Evaluation: Hybrid nanocurcumin was
evaluated by intracellular uptake and MTT assays. This study shows
robust uptake of HNC within 1 hour in pancreatic cancer cell,
MiaPaCa cells as shown in FIG. 8. Intracellular uptake of
nanoparticle was determined in pancreatic, prostate and breast
cancer cells using a Confocal Laser Scanning Microscope (CLSM). For
these studies, cells were placed on a cover slip in a 6-well tissue
culture plate and incubated at 37.degree. C. until they reached
sub-confluent levels. The cells were then exposed to 100 .mu.g/ml
concentrations of fluorescent nile red labeled hybrid
nanoparticles. After 2 hrs of incubation, cells were viewed under
the microscope.
[0084] MTT Assay: This assay was carried out in pancreatic cancer
cell line, MiaPaCa. The IC.sub.50 for the HNC formulation was found
to be at 22 .mu.M concentration (FIG. 10). To determine the effect
of hybrid nanoparticles on cell growth, cell viability (MTT) assay
was carried out in pancreatic prostate and breast cancer cell
lines. The inhibition in cell growth was measured by the MTT assay.
For this assay, 2000 cells/well were plated in a 96-well plate and
were treated with different .mu.M concentrations of free drug and
equivalent doses of drug-loaded hybrid nanoparticles. The assay was
terminated after 48 and 72 hours and relative growth inhibition
compared to control cells was measured. All studies were set up in
triplicates and repeated twice for statistical analysis. Results
were expressed as mean.+-.S.D.
[0085] Results of western blot analysis of MiaPaCa cells treated
with hybrid nanocurcumin (25 .mu.M (micromolar)); untreated; blank
nanoparticle; Curcumin (24 hrs); HNC (24 hrs) and; HNC (48 hrs) are
provided in FIG. 9.
Example II
[0086] Evaluation of the effects of Liposoma-PLGA curcumin on the
human potassium channel using human embryonic kidney (HEK) 293
cells transfected with a human ether-a-gogo-related gene (hERG):
The example deals with quantifying the in vitro effects of
Liposoma-PLGA curcumin on the potassium-selective IKr current
generated in normoxic conditions in stably transfected HEK 293
cells. The hERG assay is used to assess the potential of a compound
to interfere with the rapidly activating delayed-rectifier
potassium channel; and is based on current International Conference
on Harmonisation (ICH) Harmonized Tripartite Guidelines [ICH S7a/b]
and generally accepted procedures for the testing of pharmaceutical
compounds.
[0087] Study outline: Test articles: Batch A, Batch B, Batch C,
Batch D and Batch E. Test System: hERG-expressing HEK 293
transfected cell line. Test performed: Whole-cell patch-clamp
current acquisition and analysis. Study Temperature: 35+/-2.degree.
C.
[0088] Application of test articles, positive control and vehicle:
5 minutes of exposure to each concentration in presence of closed
circuit perfusion (2 mL/min). 5 minutes for washout period in
presence of a flow-through perfusion (2 mL/min) in addition to a
closed circuit perfusion (2 mL/min). The positive control (100 nM
E-4031) was added to naive cells obtained from the same cell line
and same passage for a period of 5 minutes in presence of a closed
circuit perfusion (2 mL/min).
[0089] Cells were under continuous stimulation of the pulses
protocol throughout the studies and cell currents were recorded
after 5 minutes of exposure to each condition.
[0090] Original data acquisition design is shown in FIG. 11.
[0091] Design for acquisition when testing the test articles or
vehicle: [0092] 1 recording made in baseline condition [0093] 1
recording made in the presence of concentration 1, 2 or 3 [0094] 1
recording made after washout (only after the concentration 3)
[0095] Design for acquisition when testing the positive control:
[0096] 1 recording made in baseline condition [0097] 1 recording
made in the presence of the positive control [0098] n=number of
responsive cells patched on which the whole protocol above could be
applied
[0099] Statistical analysis: Statistical comparisons were made
using paired Student's t-tests. For the test articles, the currents
recorded after exposure to the different test article
concentrations were statistically compared to the currents recorded
in baseline conditions. Currents recorded after the washout were
statistically compared to the currents measured after the highest
concentration of test articles. In the same way, currents recorded
after the positive control were compared to the currents recorded
in baseline conditions.
[0100] Differences were considered significant when
p.ltoreq.0.05.
[0101] Exclusion Criteria: [0102] 1. Timeframe of drug exposure not
respected [0103] 2. Instability of the seal [0104] 3. No tail
current generated by the patched cell [0105] 4. No significant
effect of the positive control [0106] 5. More than 10% variability
in capacitance transient amplitude over the duration of the
study.
[0107] Effect of the Test Articles on Whole-Cell IKr hERG
Currents:
[0108] Whole-cell currents elicited during a voltage pulse were
recorded in baseline conditions and following the application of
the selected concentrations of test articles. Currents were also
recorded following a washout period. The cells were depolarized for
one second from the holding potential (-80 mV) to a maximum value
of +40 mV, starting at -40 mV and progressing in 10 mV increments.
The membrane potential was then repolarized to -55 mV for one
second, and finally returned to -80 mV.
[0109] Whole-cell tail current amplitude was measured at a holding
potential of -55 mV, following activation of the current from -40
to +40 mV. Current amplitude was measured at the maximum (peak) of
this tail current. Current density was obtained by dividing current
amplitude by cell capacitance measured prior to capacitive
transient minimization. As per protocol, 3 concentrations of each
test article were analyzed for hERG current inhibition.
[0110] Result of the studies showing hERG current density analysis
of curcumin; liposomal curcumin; and PLGA curcumin are provided in
FIGS. 6 and 7, which show hERG current density analysis of
liposomes+curcumin; and liposomes.
[0111] Current run-down and solvent effect correction. All data
points presented in this Results Report have been corrected for
solvent effect and time-dependent current run-down. Current
run-down and solvent effects were measured simultaneously by
applying the study design in test-article free conditions (hERG
external solution or DMSO) over the same time frame as was done
with the test articles. The loss in current amplitude measured
during these so-called vehicle studies (representing both solvent
effects and time-dependent run-down) was subtracted from the loss
of amplitude measured in the presence of the test articles to
isolate the effect of the test articles, apart from the effect of
the solvent and the inevitable run-down in current amplitude over
time.
[0112] This results, as shown in FIG. 11-27, quantify the effect of
Liposomal-PLGA curcumin (Batch A, Batch B, Batch C, Batch D and
Batch E) on the rapidly activating delayed-rectifier potassium
selective current (IKr) generated under normoxic conditions in
stably transfected Human Embryonic Kidney (HEK) 293 cells.
[0113] The concentrations of curcumin (6, 12 and 18 .mu.M) were
selected and reflect a range estimated to exceed the
therapeutic.
[0114] To confirm the reversal effect of the test articles, cells
exposed to the highest concentration (18 .mu.M) were subject to a
washout period of 5 minutes. The current measured after the washout
period was not statistically different when compared to the current
left after highest concentration exposure of the compounds showing
that the effect of these compounds was not reversible.
[0115] E-4031 is one of the most selective hERG inhibitors
available to date. It was selected to demonstrate the sensitivity
of the test system. Three naive HEK293-hERG cells were exposed to
100 nM E-4031. E-4031 induced a significant inhibition of 81.8% of
the current amplitude for I+20.
[0116] Sample Information: Store at -20.degree. C., and protected
from direct sunlight: [0117] 1) Batch A-- [0118] Total weight of
sample--215 mg [0119] Curcumin content--18 micro g/mg of test
sample [0120] Material used--Polymer (PLGA), Lipid (DMPC+DMPG),
Curcumin, sucrose. [0121] 2) Batch B-- [0122] Total weight of
sample--200 mg [0123] Curcumin content--6.8 micro g/mg of test
sample [0124] Material used--Polymer (PLGA), Lipid (DMPC+DMPG),
Curcumin, sucrose. [0125] 3) Batch C-- [0126] Total weight of
sample--200 mg [0127] Curcumin content--18.2 micro g/mg of test
sample [0128] Material used--Polymer (PLGA), Chitosan, Polyvinyl
alcohol (PVA), Lipid (DMPC+DMPG), Curcumin, sucrose. [0129] 4)
Batch D--Pure curcumin [0130] Total weight--50 mg. [0131] 5) Batch
E--Liposomal curcumin [0132] Total volume--5 ml [0133] Curcumin
content--6.4 mg/ml [0134] Material used--Lipid (DMPC+DMPG),
Curcumin [0135] Molecular weight information: [0136] Curcumin
Molecular weight--368.38 g/mol [0137] PLGA (50:50)--Molecular
weight--124 kDa [0138] DMPC (PC (14:0/14:0))--Molecular
weight--677.933 g/mol [0139] DMPG--Molecular weight--688.845 g/mol
[0140] Sucrose--Molecular Weight 342.30 g/mol [0141] Chitosan--Low
Molecular weight--75-85% deacetylated [0142] Polyvinyl alcohol
(PVA)--Average molecular weight--30,000-70,000.
[0143] It is contemplated that any embodiment discussed in this
specification can be implemented with respect to any method, kit,
reagent, or composition of the invention, and vice versa.
Furthermore, compositions of the invention can be used to achieve
methods of the invention.
[0144] Evaluation of the effects of Curcumin ER and liposomal
curcumin on H-460 and A-549 lung cancer mouse xenograft model.
[0145] The purpose of this study was to quantify the mean tumor
volume of the mouse xenograft model over duration of the treatment.
Specifically, the encapsulated and liposomally coated Curcumin ER
and Liposomal Curcumin were tested using the cell lines H-460 and
A-549, lung cancer xenograft model. Briefly, Female Hsd:athymic
Nude-Foxn1nu mice 3-4 weeks old were obtained from Harlan
Laboratories, USA. The cancer cells were injected into the mice and
tumor volume was evaluated. The liposomal curcumins, Curcumin ER
and Liposomal Curcumin, were administered via subcutaneous
injection at a dose of 20 mg/kg body weight once in a week.
[0146] FIG. 28 shows the results of the treatment of the H-460
breast cancer cell line in the Hsd:Athymic Nude-Foxn1nu mice. FIG.
29 shows additional results of the treatment of the A-549 breast
cancer cell line in the Hsd:athymic Nude-Foxn1nu mice.
[0147] It will be understood that particular embodiments described
herein are shown by way of illustration and not as limitations of
the invention. The principal features of this invention can be
employed in various embodiments without departing from the scope of
the invention. Those skilled in the art will recognize, or be able
to ascertain using no more than routine experimentation, numerous
equivalents to the specific procedures described herein. Such
equivalents are considered to be within the scope of this invention
and are covered by the claims.
[0148] All publications and patent applications mentioned in the
specification are indicative of the level of skill of those skilled
in the art to which this invention pertains. All publications and
patent applications are herein incorporated by reference to the
same extent as if each individual publication or patent application
was specifically and individually indicated to be incorporated by
reference.
[0149] The use of the word "a" or "an" when used in conjunction
with the term "comprising" in the claims and/or the specification
may mean "one," but it is also consistent with the meaning of "one
or more," "at least one," and "one or more than one." The use of
the term "or" in the claims is used to mean "and/or" unless
explicitly indicated to refer to alternatives only or the
alternatives are mutually exclusive, although the disclosure
supports a definition that refers to only alternatives and
"and/or." Throughout this application, the term "about" is used to
indicate that a value includes the inherent variation of error for
the device, the method being employed to determine the value, or
the variation that exists among the study subjects.
[0150] As used in this specification and claim(s), the words
"comprising" (and any form of comprising, such as "comprise" and
"comprises"), "having" (and any form of having, such as "have" and
"has"), "including" (and any form of including, such as "includes"
and "include") or "containing" (and any form of containing, such as
"contains" and "contain") are inclusive or open-ended and do not
exclude additional, unrecited elements or method steps.
[0151] The term "or combinations thereof" as used herein refers to
all permutations and combinations of the listed items preceding the
term. For example, "A, B, C, or combinations thereof" is intended
to include at least one of: A, B, C, AB, AC, BC, or ABC, and if
order is important in a particular context, also BA, CA, CB, CBA,
BCA, ACB, BAC, or CAB. Continuing with this example, expressly
included are combinations that contain repeats of one or more item
or term, such as BB, AAA, AB, BBC, AAABCCCC, CBBAAA, CABABB, and so
forth. The skilled artisan will understand that typically there is
no limit on the number of items or terms in any combination, unless
otherwise apparent from the context. In certain embodiments, the
present invention may also include methods and compositions in
which the transition phrase "consisting essentially of" or
"consisting of" may also be used.
[0152] All of the compositions and/or methods disclosed and claimed
herein can be made and executed without undue experimentation in
light of the present disclosure. While the compositions and methods
of this invention have been described in terms of preferred
embodiments, it will be apparent to those of skill in the art that
variations may be applied to the compositions and/or methods and in
the steps or in the sequence of steps of the method described
herein without departing from the concept, spirit and scope of the
invention. All such similar substitutes and modifications apparent
to those skilled in the art are deemed to be within the spirit,
scope and concept of the invention as defined by the appended
claims.
* * * * *