U.S. patent application number 12/704426 was filed with the patent office on 2010-06-10 for nanoparticulate polycosanol formulations & novel polycosanol combinations.
This patent application is currently assigned to Elan Pharma International Ltd.. Invention is credited to Eugene R. COOPER, Laura Kline, Gary G. Liversidge, Niels P. Ryde.
Application Number | 20100143962 12/704426 |
Document ID | / |
Family ID | 29736318 |
Filed Date | 2010-06-10 |
United States Patent
Application |
20100143962 |
Kind Code |
A1 |
COOPER; Eugene R. ; et
al. |
June 10, 2010 |
NANOPARTICULATE POLYCOSANOL FORMULATIONS & NOVEL POLYCOSANOL
COMBINATIONS
Abstract
The invention provides a culture device comprising a plurality
of culture units, wherein each unit comprises a culture chamber, an
inlet port for liquid supply of the culture and an outlet port for
discharging liquid from the unit, wherein the inlet port is in
fluid communication with the culture chamber and the culture
chamber is in fluid communication with the outlet port for allowing
a liquid flow through the culture chamber. The culture device is
particularly suitable for testing immune cells and immunofunction
in vitro. Aspects of the invention include a culture device and
associated methods for cultivating immune cells and an in vitro
method of analysing the effect of a test compound on immune
cells.
Inventors: |
COOPER; Eugene R.; (Berwyn,
PA) ; Kline; Laura; (Harleysville, PA) ;
Liversidge; Gary G.; (West Chester, PA) ; Ryde; Niels
P.; (Malvern, PA) |
Correspondence
Address: |
Elan Drug Delivery, Inc. c/o Foley & Lardner
3000 K Street, N.W., Suite 500
Washington
DC
20007-5109
US
|
Assignee: |
Elan Pharma International
Ltd.
|
Family ID: |
29736318 |
Appl. No.: |
12/704426 |
Filed: |
February 11, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10457811 |
Jun 10, 2003 |
|
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12704426 |
|
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60387463 |
Jun 10, 2002 |
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Current U.S.
Class: |
435/29 ;
435/289.1; 435/297.1; 435/372 |
Current CPC
Class: |
A61K 31/045 20130101;
A23L 33/105 20160801; A61P 9/00 20180101; A61K 45/06 20130101; A61P
25/28 20180101; A61P 9/10 20180101; A61P 3/06 20180101; A23L 33/10
20160801; A61K 31/56 20130101; A61K 9/145 20130101 |
Class at
Publication: |
435/29 ;
435/289.1; 435/297.1; 435/372 |
International
Class: |
C12N 5/078 20100101
C12N005/078; C12M 3/00 20060101 C12M003/00; C12M 3/06 20060101
C12M003/06; C12Q 1/02 20060101 C12Q001/02 |
Claims
1. A culture device having a top side, a bottom side, and at least
one lateral side, comprising a plurality of culture units, wherein
each unit comprises (i) a culture chamber, (ii) an inlet port for
reversibly connecting the unit with an external liquid supply and
(iii) an outlet port for discharging liquid from the unit, wherein
the inlet port is in fluid communication with the culture chamber
and the culture chamber is in fluid communication with the outlet
port for allowing a liquid flow through the culture chamber,
wherein the inlet port is accessible for connecting an external
liquid supply from the top side or lateral side of the culture
device, and the outlet port is accessible for connecting a
discharge conduct from the top side or lateral side of the
device.
2. The culture device of claim 1, wherein the mainstream of the
liquid flow traverses the culture chamber within the plane of the
culture device.
3. The culture device according to claim 1, wherein the fluid
communication is established by a channel between the culture
chamber and the inlet port and a channel between the culture
chamber and the outlet port.
4. The culture device according to claim 1, wherein the culture
chamber or chambers: (a) are sealed on at least one side with a
gas-permeable foil; (b) are gas-tightly closed except for openings
involved in fluid communication of the culture chamber with the
ports; (c) have a culture volume from 25 to 1000 .mu.l; (d) are
translucent for allowing microscopic inspection of cells present in
the culture chambers; (e) comprises a preformed matrix; (f)
comprises living eukaryotic cell material embedded in a matrix; (g)
comprises living eukaryotic cell material seeded on a matrix; or
(h) any combination thereof.
5. (canceled)
6. The culture device according to claim 5, further comprising a
conduct between the inlet port and the external liquid supply, the
conduct comprising a gas permeable conduct that allows
equilibrating liquid media to a predefined oxygen and carbon
dioxide content.
7. (canceled)
8. The culture device according to claim 4, wherein the culture
chamber has a culture volume from 50 to 250 .mu.l or from, 50 to
100 .mu.l.
9. The culture device according to claim 1, the device comprising 2
to 200 culture units.
10. The culture device of claim 1, wherein test substances and/or
stimulatory agents may be added to living cell material in each
individual unit separately.
11. (canceled)
12. (canceled)
13. (canceled)
14. (canceled)
15. The culture device according to claim 4, wherein the matrix is
selected from a hydrogel, foam or non-woven fibres.
16. (canceled)
17. The culture device according to claim 4, wherein the living
eukaryotic cell material comprises leucocytes or co-cultures of
leucocytes with other cells of interest.
18. The culture device according to claim 17, wherein the
leucocytes are whole peripheral blood mononuclear cells, defined
subpopulations of peripheral blood mononuclear cells, in vitro
differentiated peripheral blood mononuclear cell subpopulations and
any co-cultures of these.
19. The culture device according to claim 17, wherein the other
cells of interest are selected from the group consisting of
endothelial cells, stem cells, follicular dendritic cells, stromal
cells and others.
20. The culture device according to claim 1, any wherein: (a) the
culture unit comprises an additional inlet port for introducing a
suspension comprising eukaryotic cells and/or an aqueous
matrix-forming composition into the culture chamber; (b) the
culture units further comprise hollow fibre membranes traversing
the culture cavity, whereby the hollow fibre membranes are in fluid
communication with the culture chamber and the ports for allowing a
liquid flow through the culture chamber; or (c) any combination
thereof.
21. (canceled)
22. The culture device according to claim 20, wherein the hollow
fibre membranes are mounted in a separate cassette comprising the
culture chamber.
23. The culture device according to claim 1, that provides a
microenvironment.
24. A method of cultivating immune cells, comprising the steps of
(a) introducing a suspension of immune cells in an aqueous
matrix-forming composition into a culture chamber, (b) solidifying
the suspension, thereby forming a matrix having the immune cells
embedded therein, and (c) incubating the immune cells at predefined
culture conditions for a predefined period of time, whereby the
matrix is continuously perfused with liquid culture media and
supplements.
25. A method of cultivating immune cells, comprising the steps of:
(a) introducing a suspension of immune cells in an aqueous
matrix-forming composition into the culture chamber of at least one
culture unit of the culture device according to claim 1, (b)
solidifying the suspension, thereby forming a matrix having the
immune cells embedded therein, and (c) incubating the immune cells
at predefined culture conditions for a predefined period of time,
whereby the matrix is continuously perfused with liquid culture
media and supplements.
26. A method of cultivating immune cells, comprising incubating
immune cells embedded in a matrix at predefined culture conditions
for a predefined period of time, whereby the matrix is continuously
perfused with liquid culture media and supplements.
27. A method of cultivating immune cells, comprising incubating
immune cells embedded in a matrix in the culture chamber of at
least one culture unit of the culture device according to claim 1,
at predefined culture conditions for a predefined period of time,
whereby the matrix is continuously perfused with liquid culture
media and supplements.
28. The method according to claim 25, wherein the individual
culture units provide comparable conditions.
29. The method according to claim 24, wherein the matrix is
selected from a hydrogel, foam, sponge or non-woven fibres.
30. (canceled)
31. The method according to claim 24, wherein the immune cells are
leucocytes or co-cultures of leucocytes with other cells of
interest.
32. The method according to claim 31, wherein: (a) the leucocytes
can be whole peripheral blood mononuclear cells, defined
subpopulations of peripheral blood mononuclear cells, in vitro
differentiated peripheral blood mononuclear cell subpopulations and
any co-cultures of these; (b) the other cells of interest are
selected from the group consisting of endothelial cells, stem
cells, follicular dendritic cells, stromal cells and others; or (c)
any combination thereof.
33. (canceled)
34. The method according to claim 24, wherein: (a) the immune cells
are incubated at predefined culture conditions for a period of time
of up to 8 weeks; (b) the culture conditions are defined by culture
media, supplements, matrices, technically supported
micro-environment and gas supply; (c) cultured cells are provided
with gas through a gas permeable membrane that seals at least one
side of the culture chamber; (d) the matrix, culture media
composition, cell density and cell mixture allow formation of
micro-organoid structures; (e) the matrix, culture media
composition, cell density and cell mixture allow formation of a
microenvironment; or (f) any combination thereof
35. (canceled)
36. (canceled)
37. (canceled)
38. (canceled)
39. A method of analysing the effect of a test compound on immune
cells, which comprises the steps of (a) incubating immune cells
embedded in a matrix in the culture chamber of at least one culture
unit of the culture device according to claim 1, at predefined
culture conditions for a predefined period of time, whereby the
matrix is continuously perfused with liquid culture media and
supplements, (b) adding the test compound to at least one culture
unit through the inlet port, (c) incubating immune cells as
described in step a) in the presence of the at least one test
compound (d) removing at least one sample of the culture media
discharged from the outlet port of the at least one culture unit to
which the test compound was added, and (e) analysing the sample for
an effect of the test compound on the immune cells.
40. The method according to claim 39, wherein the matrix is
selected from a hydrogel, foam, sponge or non-woven fibres.
41. (canceled)
42. (canceled)
43. The method according to claim 39, wherein: (a) the sample is
removed while maintaining incubation of the cells; (b) in step e)
the presence or concentration of soluble factors selected from
cytokines, chemokines and antibodies are analysed in the sample;
(c) the test compound, concentration of test compound, time of
exposing the cells to the test compound are different between
individual culture units; (d) further comprising analysis of the
immune cells by a method selected from the group of immune
fluorescence, light microscopy, flow cytometry, fluorescence
activated cell sorting, histology, molecular biology techniques and
enzyme linked immuno-spot methods after the culture process; or (e)
any combination thereof.
44. (canceled)
45. The method according to claims 25, which further comprises
online monitoring and/or offline monitoring of cell culture
conditions in culture media discharged at the outlet port.
46. The method according to claim 45, wherein the cell culture
conditions comprise oxygen content, pH and metabolites.
47. (canceled)
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Divisional of U.S. patent application
Ser. No. 10/457,811, filed on Jun. 10, 2003, which claims priority
to U.S. Provisional Patent Application No. 60/387,463, filed on
Jun. 10, 2002. The contents of these applications are incorporated
herein by reference in their entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to nanoparticulate
compositions comprising at least one polycosanol and novel
polycosanol combinations. The nanoparticulate polycosanol particles
preferably have an effective average particle size of less than
about 2000 nm. In another aspect, this invention includes novel
combinations of polycosanols and other cholesterol lowering agents
and methods of using the same.
BACKGROUND OF THE INVENTION
I. Background Regarding Nanoparticulate Active Agent
Compositions
[0003] Nanoparticulate active agent compositions, first described
in U.S. Pat. No. 5,145,684 ("the '684 patent"), are particles
consisting of a poorly soluble therapeutic or diagnostic agent
having adsorbed onto, or associated with, the surface thereof a
non-crosslinked surface stabilizer. Many factors can affect
bioavailability including the dosage form and various properties,
e.g., dissolution rate of the drug. Poor bioavailability is a
significant problem encountered in the development of
pharmaceutical compositions, particularly those containing an
active ingredient that is poorly soluble in water. By decreasing
the particle size of an active agent, the surface area of the
composition is increased, thereby generally resulting in an
increased bioavailability. The '684 patent does not teach
nanoparticulate compositions of polycosanols.
[0004] Methods of making nanoparticulate active agent compositions
are described in, for example, U.S. Pat. Nos. 5,518,187 and
5,862,999, both for "Method of Grinding Pharmaceutical Substances;"
U.S. Pat. No. 5,718,388, for "Continuous Method of Grinding
Pharmaceutical Substances;" and U.S. Pat. No. 5,510,118 for
"Process of Preparing Therapeutic Compositions Containing
Nanoparticles." None of these patents teach nanoparticulate
compositions of polycosanols.
[0005] Nanoparticulate active agent compositions are also
described, for example, in U.S. Pat. No. 5,298,262 for "Use of
Ionic Cloud Point Modifiers to Prevent Particle Aggregation During
Sterilization;" U.S. Pat. No. 5,302,401 for "Method to Reduce
Particle Size Growth During Lyophilization;" U.S. Pat. No.
5,318,767 for "X-Ray Contrast Compositions Useful in Medical
Imaging;" U.S. Pat. No. 5,326,552 for "Novel Formulation For
Nanoparticulate X-Ray Blood Pool Contrast Agents Using High
Molecular Weight Non-ionic Surfactants;" U.S. Pat. No. 5,328,404
for "Method of X-Ray Imaging Using Iodinated Aromatic
Propanedioates;" U.S. Pat. No. 5,336,507 for "Use of Charged
Phospholipids to Reduce Nanoparticle Aggregation;" U.S. Pat. No.
5,340,564 for "Formulations Comprising Olin 10-G to Prevent
Particle Aggregation and Increase Stability;" U.S. Pat. No.
5,346,702 for "Use of Non-Ionic Cloud Point Modifiers to Minimize
Nanoparticulate Aggregation During Sterilization;" U.S. Pat. No.
5,349,957 for "Preparation and Magnetic Properties of Very Small
Magnetic-Dextran Particles;" U.S. Pat. No. 5,352,459 for "Use of
Purified Surface Modifiers to Prevent Particle Aggregation During
Sterilization;" U.S. Pat. Nos. 5,399,363 and 5,494,683, both for
"Surface Modified Anticancer Nanoparticles;" U.S. Pat. No.
5,401,492 for "Water Insoluble Non-Magnetic Manganese Particles as
Magnetic Resonance Enhancement Agents;" U.S. Pat. No. 5,429,824 for
"Use of Tyloxapol as a Nanoparticulate Stabilizer;" U.S. Pat. No.
5,447,710 for "Method for Making Nanoparticulate X-Ray Blood Pool
Contrast Agents Using High Molecular Weight Non-ionic Surfactants;"
U.S. Pat. No. 5,451,393 for "X-Ray Contrast Compositions Useful in
Medical Imaging;" U.S. Pat. No. 5,466,440 for "Formulations of Oral
Gastrointestinal Diagnostic X-Ray Contrast Agents in Combination
with Pharmaceutically Acceptable Clays;" U.S. Pat. No. 5,470,583
for "Method of Preparing Nanoparticle Compositions Containing
Charged Phospholipids to Reduce Aggregation;" U.S. Pat. No.
5,472,683 for "Nanoparticulate Diagnostic Mixed Carbamic Anhydrides
as X-Ray Contrast Agents for Blood Pool and Lymphatic System
Imaging;" U.S. Pat. No. 5,500,204 for "Nanoparticulate Diagnostic
Dimers as X-Ray Contrast Agents for Blood Pool and Lymphatic System
Imaging;" U.S. Pat. No. 5,518,738 for "Nanoparticulate NSAID
Formulations;" U.S. Pat. No. 5,521,218 for "Nanoparticulate
Iododipamide Derivatives for Use as X-Ray Contrast Agents;" U.S.
Pat. No. 5,525,328 for "Nanoparticulate Diagnostic Diatrizoxy Ester
X-Ray Contrast Agents for Blood Pool and Lymphatic System Imaging;"
U.S. Pat. No. 5,543,133 for "Process of Preparing X-Ray Contrast
Compositions Containing Nanoparticles;" U.S. Pat. No. 5,552,160 for
"Surface Modified NSAID Nanoparticles;" U.S. Pat. No. 5,560,931 for
"Formulations of Compounds as Nanoparticulate Dispersions in
Digestible Oils or Fatty Acids;" U.S. Pat. No. 5,565,188 for
"Polyalkylene Block Copolymers as Surface Modifiers for
Nanoparticles;" U.S. Pat. No. 5,569,448 for "Sulfated Non-ionic
Block Copolymer Surfactant as Stabilizer Coatings for Nanoparticle
Compositions;" U.S. Pat. No. 5,571,536 for "Formulations of
Compounds as Nanoparticulate Dispersions in Digestible Oils or
Fatty Acids;" U.S. Pat. No. 5,573,749 for "Nanoparticulate
Diagnostic Mixed Carboxylic Anydrides as X-Ray Contrast Agents for
Blood Pool and Lymphatic System Imaging;" U.S. Pat. No. 5,573,750
for "Diagnostic Imaging X-Ray Contrast Agents;" U.S. Pat. No.
5,573,783 for "Redispersible Nanoparticulate Film Matrices With
Protective Overcoats;" U.S. Pat. No. 5,580,579 for "Site-specific
Adhesion Within the GI Tract Using Nanoparticles Stabilized by High
Molecular Weight, Linear Poly(ethylene Oxide) Polymers;" U.S. Pat.
No. 5,585,108 for "Formulations of Oral Gastrointestinal
Therapeutic Agents in Combination with Pharmaceutically Acceptable
Clays;" U.S. Pat. No. 5,587,143 for "Butylene Oxide-Ethylene Oxide
Block Copolymers Surfactants as Stabilizer Coatings for
Nanoparticulate Compositions;" U.S. Pat. No. 5,591,456 for "Milled
Naproxen with Hydroxypropyl Cellulose as Dispersion Stabilizer;"
U.S. Pat. No. 5,593,657 for "Novel Barium Salt Formulations
Stabilized by Non-ionic and Anionic Stabilizers;" U.S. Pat. No.
5,622,938 for "Sugar Based Surfactant for Nanocrystals;" U.S. Pat.
No. 5,628,981 for "Improved Formulations of Oral Gastrointestinal
Diagnostic X-Ray Contrast Agents and Oral Gastrointestinal
Therapeutic Agents;" U.S. Pat. No. 5,643,552 for "Nanoparticulate
Diagnostic Mixed Carbonic Anhydrides as X-Ray Contrast Agents for
Blood Pool and Lymphatic System Imaging;" U.S. Pat. No. 5,718,388
for "Continuous Method of Grinding Pharmaceutical Substances;" U.S.
Pat. No. 5,718,919 for "Nanoparticles Containing the R(-)Enantiomer
of Ibuprofen;" U.S. Pat. No. 5,747,001 for "Aerosols Containing
Beclomethasone Nanoparticle Dispersions;" U.S. Pat. No. 5,834,025
for "Reduction of Intravenously Administered Nanoparticulate
Formulation Induced Adverse Physiological Reactions;" U.S. Pat. No.
6,045,829 "Nanocrystalline Formulations of Human Immunodeficiency
Virus (HIV) Protease Inhibitors Using Cellulosic Surface
Stabilizers;" U.S. Pat. No. 6,068,858 for "Methods of Making
Nanocrystalline Formulations of Human Immunodeficiency Virus (HIV)
Protease Inhibitors Using Cellulosic Surface Stabilizers;" U.S.
Pat. No. 6,153,225 for "Injectable Formulations of Nanoparticulate
Naproxen;" U.S. Pat. No. 6,165,506 for "New Solid Dose Form of
Nanoparticulate Naproxen;" U.S. Pat. No. 6,221,400 for "Methods of
Treating Mammals Using Nanocrystalline Formulations of Human
Immunodeficiency Virus (HIV) Protease Inhibitors;" U.S. Pat. No.
6,264,922 for "Nebulized Aerosols Containing Nanoparticle
Dispersions;" U.S. Pat. No. 6,267,989 for "Methods for Preventing
Crystal Growth and Particle Aggregation in Nanoparticle
Compositions;" U.S. Pat. No. 6,270,806 for "Use of PEG-Derivatized
Lipids as Surface Stabilizers for Nanoparticulate Compositions;"
U.S. Pat. No. 6,316,029 for "Rapidly Disintegrating Solid Oral
Dosage Form," U.S. Pat. No. 6,375,986 for "Solid Dose
Nanoparticulate Compositions Comprising a Synergistic Combination
of a Polymeric Surface Stabilizer and Dioctyl Sodium
Sulfosuccinate;" U.S. Pat. No. 6,428,814 for "Bioadhesive
Nanoparticulate Compositions Having Cationic Surface Stabilizers;"
U.S. Pat. No. 6,431,478 for "Small Scale Mill;" and U.S. Pat. No.
6,432,381 for "Methods for Targeting Drug Delivery to the Upper
and/or Lower Gastrointestinal Tract," all of which are specifically
incorporated by reference. In addition, U.S. Patent Application No.
20020012675 A1, published on Jan. 31, 2002, for "Controlled Release
Nanoparticulate Compositions," describes nanoparticulate
compositions, and is specifically incorporated by reference. None
of these patents teach nanoparticulate compositions of
polycosanols.
[0006] Amorphous small particle compositions are described, for
example, in U.S. Pat. No. 4,783,484 for "Particulate Composition
and Use Thereof as Antimicrobial Agent;" U.S. Pat. No. 4,826,689
for "Method for Making Uniformly Sized Particles from
Water-Insoluble Organic Compounds;" U.S. Pat. No. 4,997,454 for
"Method for Making Uniformly-Sized Particles From Insoluble
Compounds;" U.S. Pat. No. 5,741,522 for "Ultrasmall, Non-aggregated
Porous Particles of Uniform Size for Entrapping Gas Bubbles Within
and Methods;" and U.S. Pat. No. 5,776,496, for "Ultrasmall Porous
Particles for Enhancing Ultrasound Back Scatter."
II. Background Regarding Polycosanols
[0007] Policosanol (polycosanol) is a complex mixture of
concentrated n-alkyl alcohols derived from sugar cane and the wax
of honey bees, among others. The polycosanols are extracted by
known methods. These active substances act to lower cholesterol
levels by several mechanisms, including blocking the formation of
cholesterol in the liver. Well designed clinical trials have
included short and long-term, randomized double-blind studies
comparing policosanol to a placebo, as well as blind comparative
trials versus statin drugs, fibrates, niacin and probucol. The
results from the clinical studies involving nearly 30,000 total
patients demonstrated that policosanol is one of the best known
answers for fighting cholesterol levels and actively lowering them
safely and effectively.
[0008] Policosanol (polycosanol) produces cholesterol lowering
effects within the first 6-8 weeks of use. At a daily dosage of 10
mg taken at night, LDL cholesterol levels typically drop by 20-25%
within the first six months of use. At a dosage of 20 mg, LDL
levels typically drop by 25-30%. HDL levels typically increase by
15-25% only after two months of use. The combined LDL reduction and
HDL increase will produce a significant and dramatic improvement in
the LDL to HDL ratio. See
http://www.firstratemall.com/cholesterolfreeheart/.
[0009] Cholesterol is transported through the bloodstream by
special molecules called lipoproteins. There are three main kinds
of lipoproteins: high-density lipoprotein (HDL), low-density
lipoprotein (LDL), and very low-density lipoprotein (VLDL). LDL
carries fats from the liver to the body cells, while HDL carries
fat back to the liver. The higher the level of LDL the greater the
risk of fat-related illnesses such as atherosclerosis. In contrast,
HDL protects against these illnesses because it removes fats from
circulation and puts them back into storage in the liver.
[0010] The fatty acids in policosanol are primarily 1-Octacosanol,
1-Triacontanol, 1-Tetracosanol, and 1-Hexacosanol. Typical usage
levels range from 500 -10,000 micrograms per serving/dose. Typical
commercially available commercial compositions are 90% minimum
fatty alcohols of (a) 1-Tetracosanol: 0-10%; (b) 1-Hexacosanol:
2-15%; (c) 1-Heptacosanol: 0-0.5%; (d) 1-Octacosanol: 55-70%; (e)
1-Nonacosanol: 0-10%; (f) 1-Triacontanol: 5-20%; (g)
1-Dotriacontanol: 0.1-10%; and (h) 1-Tetratriacontanol:
0.1-10%.
[0011] It would be desirable to provide stable, dispersible
polycosanol particles, up to about the 2000 nm size range, which
can be readily prepared and formulated in pharmaceutically useful
and more convenient, palatable forms for consumption. The present
invention satisfies these needs.
SUMMARY OF THE INVENTION
[0012] The present invention relates to nanoparticulate active
agent compositions comprising at least one polycosanol and novel
polycosanol combinations. The compositions preferably comprise at
least one polycosanol and at least one surface stabilizer adsorbed
on or associated with the surface of the one or more polycosanol
particles. The nanoparticulate polycosanol particles preferably
have an effective average particle size of less than about 2000
nm.
[0013] Another aspect of the invention is directed to
pharmaceutical compositions comprising a nanoparticulate
polycosanol composition of the invention. The pharmaceutical
compositions preferably comprise at least one polycosanol, at least
one surface stabilizer, and at least one pharmaceutically
acceptable carrier, as well as any desired excipients known to
those in the art and formulated into the dosage form desired.
[0014] In another aspect of this invention, novel combinations of
polycosanols and at least one other cholesterol lowering agent are
described and methods of using the same are taught.
[0015] Another aspect of the invention is directed to a
nanoparticulate polycosanol composition having improved
pharmacokinetic profiles as compared to conventional
microcrystalline polycosanol formulations, such as improved
T.sub.max, C.sub.max, and/or AUC parameters.
[0016] One embodiment of the invention encompasses a polycosanol
composition, wherein the pharmacokinetic profile of the polycosanol
is not affected by the fed or fasted state of a subject ingesting
the composition, preferably as defined by C.sub.max and AUC
guidelines given by the U.S. Food and Drug Administration and/or
the corresponding European regulatory agency (EMEA).
[0017] In yet another embodiment, the invention encompasses a
polycosanol composition of the invention, wherein administration of
the composition to a subject in a fasted state is bioequivalent to
administration of the composition to a subject in a fed state, in
particular as defined by C.sub.max and AUC guidelines given by the
U.S. Food and Drug Administration and the corresponding European
regulatory agency (EMEA).
[0018] Other embodiments of the invention include, but are not
limited to, nanoparticulate polycosanol compositions which, as
compared to conventional non-nanoparticulate formulations of the
same polycosanol, preferably have one or more of the following
properties: (1) smaller tablet or other solid dosage form size; (2)
smaller doses of drug required to obtain the same pharmacological
effect; (3) increased bioavailability; (4) an increased rate of
dissolution for the nanoparticulate polycosanol compositions; and
(6) bioadhesive polycosanol compositions. This invention further
discloses a method of making a nanoparticulate polycosanol
composition according to the invention. Such method comprises
contacting at least one polycosanol with at least one surface
stabilizer for a time and under conditions sufficient to provide a
nanoparticulate polycosanol composition. The one or more surface
stabilizers can be contacted with the polycosanol before,
preferably during, or after size reduction of the polycosanol.
[0019] The present invention is also directed to methods of
treatment using the nanoparticulate polycosanol compositions of the
invention for conditions such as hypercholesterolemia,
hypertriglyceridemia, coronary heart disease, and peripheral
vascular disease (including symptomatic carotid artery disease). In
one aspect, the compositions of the invention can be used as
adjunctive therapy to diet for the reduction of LDL-C, total-C,
triglycerides, and Apo B in adult patients with primary
hypercholesterolemia or mixed dyslipidemia (Fredrickson Types IIa
and IIb). In another aspect, the compositions can be used as
adjunctive therapy to diet for treatment of adult patients with
hypertriglyceridemia (Fredrickson Types IV and V hyperlipidemia).
Markedly elevated levels of serum tryglycerides (e.g., >2000
mg/dL) may increase the risk of developing pancreatitis. Other
diseases that may be directly or indirectly associated with
elevated, uncontrolled cholesterol metabolism, e.g., restenosis and
Alzheimer's disease, may also be treated with the compositions of
this invention. Other methods of treatment using the
nanoparticulate polycosanol compositions of the present invention
are known to those of skill in the art.
[0020] Such methods comprise administering to a subject a
therapeutically effective amount of a nanoparticulate polycosanol
pharmaceutical composition according to the invention.
[0021] Both the foregoing general description and the following
detailed description are exemplary and explanatory and are intended
to provide further explanation of the invention as claimed. Other
objects, advantages, and novel features will be readily apparent to
those skilled in the art from the following detailed description of
the invention.
DETAILED DESCRIPTION OF THE INVENTION
[0022] The present invention relates to nanoparticulate active
agent compositions comprising at least one polycosanol and novel
polycosanol combinations. The compositions preferably comprise at
least one polycosanol and at least one surface stabilizer adsorbed
on or associated with the surface of the polycosanol particles. The
nanoparticulate polycosanol particles preferably have an effective
average particle size of less than about 2000 nm.
[0023] As taught in the '684 patent, not every combination of
surface stabilizer and active agent will result in a stable
nanoparticulate composition. It was surprisingly discovered that
stable nanoparticulate polycosanol compositions can be made.
[0024] A need exists for safer and higher potency polycosanols.
Compositions of nanoparticulate polycosanols decrease the amount of
drug needed and this, in turn, decreases adverse side effects while
providing maximum dose response. Additionally, a longer plasma
half-life is believed to be associated with nanoparticulate
polycosanol compositions of the invention. Moreover, increasing the
duration of effect of the polycosanol compositions is expected to
result in even lower serum cholesterol levels, with a further
reduction in dose expected.
[0025] In general, the rate of dissolution of a particulate drug
can increase with increasing surface area, e.g., decreasing
particle size. Consequently, methods of making finely divided drugs
have been studied and efforts have been made to control the size
and size range of drug particles in pharmaceutical compositions.
However, nanoparticulate active agent formulations suitable for
administration as a pharmaceutical require formulation of the
active ingredient into a colloidal dispersion exhibiting the
acceptable nanoparticle size range and the stability to maintain
such size range and not agglomerate. Merely increasing surface area
by decreasing particle size does not assure success. Further
challenges include forming solid dose forms redispersible into the
nanoparticle form upon administration to the patient to maintain
the benefit of the nanoparticle polycosanol over the traditional
dosage form.
[0026] Advantages of the nanoparticulate polycosanol compositions
of the invention as compared to conventional non-nanoparticulate
formulations of the same polycosanol preferably include, but are
not limited to: (1) smaller tablet or other solid dosage form size;
(2) smaller doses of drug required to obtain the same
pharmacological effect; (3) increased bioavailability; (4)
substantially similar pharmacokinetic profiles of the
nanoparticulate polycosanol compositions when administered in the
fed versus the fasted state; (5) improved pharmacokinetic profiles;
(6) bioequivalency of the nanoparticulate polycosanol compositions
when administered in the fed versus the fasted state; (7) an
increased rate of dissolution for the nanoparticulate polycosanol
compositions; (8) bioadhesive polycosanol compositions; and (9) the
nanoparticulate polycosanol compositions can be used in conjunction
with other active agents.
[0027] The present invention also includes nanoparticulate
polycosanol compositions together with one or more non-toxic
physiologically acceptable carriers, adjuvants, or vehicles,
collectively referred to as carriers. The compositions can be
formulated for parenteral injection (e.g., intravenous,
intramuscular, or subcutaneous), oral administration in solid,
liquid, or aerosol form, vaginal, nasal, rectal, ocular, local
(powders, ointments or drops), buccal, intracisternal,
intraperitoneal, or topical administration, and the like.
[0028] A preferred dosage form of the invention is a solid dosage
form, although any pharmaceutically acceptable dosage form can be
utilized. Exemplary solid dosage forms include, but are not limited
to, tablets, capsules, sachets, lozenges, powders, pills, or
granules. The solid dosage form can be, for example, a fast melt
dosage form, controlled release dosage form, lyophilized dosage
form, delayed release dosage form, extended release dosage form,
pulsatile release dosage form, mixed immediate release and
controlled release dosage form, or a combination thereof. A solid
dose tablet formulation is preferred.
[0029] The preferred method by which the composition of the present
invention is used to reduce cholesterol absorption includes the
step of mixing the composition with foods and beverages and mixing.
The novel food additive is also effective as an additive in
margarine, cooking oils or shortening and preferably fruit and
vegetable juices preferably orange or tomato juice for the purpose
of reducing serum cholesterol in humans who ingest food products
made with the novel composition of this invention.
[0030] The present invention is described herein using several
definitions, as set forth below and throughout the application.
[0031] "About" will be understood by persons of ordinary skill in
the art and will vary to some extent on the context in which the
term is used. If there are uses of the term which are not clear to
persons of ordinary skill in the art given the context in which it
is used, "about" will mean up to plus or minus 10% of the
particular term. "Conventional" or "non-nanoparticulate active
agent" shall mean an active agent which is solubilized or which has
an effective average particle size of greater than about 2
microns.
[0032] "Poorly water soluble drugs" as used herein means those
having a solubility of less than about 30 mg/ml, preferably less
than about 20 mg/ml, preferably less than about 10 mg/ml, or
preferably less than about 1 mg/ml. Such drugs tend to be
eliminated from the gastrointestinal tract before being absorbed
into the circulation. Moreover, poorly water soluble drugs tend to
be unsafe for intravenous administration techniques, which are used
primarily in conjunction with highly water soluble drug
substances.
[0033] As used herein with reference to stable polycosanol
particles, "stable" includes, but is not limited to, one or more of
the following parameters: (1) that the polycosanol particles do not
appreciably flocculate or agglomerate due to interparticle
attractive forces, or otherwise significantly increase in particle
size over time; (2) that the physical structure of the polycosanol
particles is not altered over time, such as by conversion from an
amorphous phase to crystalline phase; (3) that the polycosanol
particles are chemically stable; and/or (4) where the polycosanol
has not been subject to a heating step at or above the melting
point of the polycosanol in the preparation of the nanoparticles of
the invention.
[0034] "Therapeutically effective amount" as used herein with
respect to a drug dosage, shall mean that dosage that provides the
specific pharmacological response for which the drug is
administered in a significant number of subjects in need of such
treatment. It is emphasized that "therapeutically effective
amount," administered to a particular subject in a particular
instance will not always be effective in treating the diseases
described herein, even though such dosage is deemed a
`therapeutically effective amount` by those skilled in the art. It
is to be further understood that drug dosages are, in particular
instances, measured as oral dosages, or with reference to drug
levels as measured in blood.
I. Preferred Characteristics of the Polycosanol Compositions of the
Invention
[0035] A. Increased Bioavailability and Lower Dosages
[0036] The polycosanol compositions of the invention preferably
exhibit increased bioavailability, at the same dose of the same
polycosanol, require smaller doses, and show longer plasma
half-life as compared to prior conventional polycosanol
formulations. In one aspect of the invention, pharmaceutical
polycosanol compositions have enhanced bioavailability such that
the polycosanol dosage can be reduced, resulting in a decrease in
toxicity associated with such polycosanols. It has been
surprisingly found in the present invention that stable
compositions of nanoparticulate polycosanols can be formed that
permit therapeutic levels at desirably lower dosage.
[0037] Greater bioavailability of the polycosanol compositions of
the invention can enable a smaller solid dosage size. This is
particularly significant for patient populations such as the
elderly, juvenile, and infant.
[0038] B. Improved Pharmacokinetic Profiles
[0039] The invention also preferably provides polycosanol
compositions having a desirable pharmacokinetic profile when
administered to mammalian subjects. The desirable pharmacokinetic
profile of the polycosanol compositions preferably includes, but is
not limited to: (1) that the T.sub.max of a polycosanol when
assayed in the plasma of a mammalian subject following
administration is preferably less than the T.sub.max for a
conventional, non-nanoparticulate form of the same polycosanol,
administered at the same dosage; (2) that the C.sub.max of a
polycosanol when assayed in the plasma of a mammalian subject
following administration is preferably greater than the C.sub.max
for a conventional, non-nanoparticulate form of the same
polycosanol, administered at the same dosage; and/or (3) that the
AUC of a polycosanol when assayed in the plasma of a mammalian
subject following administration, is preferably greater than the
AUC for a conventional, non-nanoparticulate form of the same
polycosanol, administered at the same dosage.
[0040] The desirable pharmacokinetic profile, as used herein, is
the pharmacokinetic profile measured after the initial dose of a
polycosanol. The compositions can be formulated in any way as
described below and as known to those of skill in the art. A
preferred polycosanol composition of the invention exhibits in
comparative pharmacokinetic testing with a non-nanoparticulate
formulation of the same polycosanol, administered at the same
dosage, a T.sub.max not greater than about 90%, not greater than
about 80%, not greater than about 70%, not greater than about 60%,
not greater than about 50%, not greater than about 30%, not greater
than about 25%, not greater than about 20%, not greater than about
15%, or not greater than about 10% of the T.sub.max, exhibited by
the non-nanoparticulate formulation of the same polycosanol.
[0041] A preferred polycosanol and composition of the invention
exhibits in comparative pharmacokinetic testing with a
non-nanoparticulate formulation of the same polycosanol,
administered at the same dosage, a C.sub.max which is at least
about 10%, at least about 20%, at least about 30%, at least about
40%, at least about 50%, at least about 60%, at least about 70%, at
least about 80%, at least about 90%, or at least about 100% greater
than the C.sub.max exhibited by the non-nanoparticulate formulation
of the same polycosanol.
[0042] A preferred polycosanol composition of the invention
exhibits in comparative pharmacokinetic testing with a
non-nanoparticulate formulation of the same polycosanol,
administered at the same dosage, an AUC which is at least about
10%, at least about 20%, at least about 30%, at least about 40%, at
least about 50%, at least about 60%, at least about 70%, at least
about 80%, at least about 90%, or at least about 100% greater than
the AUC exhibited by the non-nanoparticulate formulation of the
same polycosanol.
[0043] Any formulation giving the desired pharmacokinetic profile
is suitable for administration according to the present methods.
Exemplary types of formulations giving such profiles are liquid
dispersions, gels, aerosols, ointments, creams, solid dose forms,
etc. of a nanoparticulate polycosanol.
[0044] C. The Pharmacokinetic Profiles of the Polycosanol
Compositions of the Invention are not Affected by the Fed or Fasted
State of the Subject Ingesting the Compositions
[0045] The invention encompasses polycosanol compositions wherein
the pharmacokinetic profile of the polycosanol is preferably not
substantially affected by the fed or fasted state of a subject
ingesting the composition, when administered to a human. This means
that there is no substantial difference in the quantity of drug
absorbed or the rate of drug absorption when the nanoparticulate
polycosanol compositions are administered in the fed versus the
fasted state.
[0046] The invention also encompasses a polycosanol composition in
which administration of the composition to a subject in a fasted
state is bioequivalent to administration of the composition to a
subject in a fed state. "Bioequivalency" is preferably established
by a 90% Confidence Interval (CI) of between 0.80 and 1.25 for both
C.sub.max and AUC under U.S. Food and Drug Administration
regulatory guidelines, or a 90% CI for AUC of between 0.80 to 1.25,
and a 90% CI for C.sub.max of between 0.70 to 1.43, under the
European EMEA regulatory guidelines (T.sub.max is not relevant for
bioequivalency determinations under USFDA and EMEA regulatory
guidelines).
[0047] Benefits of a dosage form which substantially eliminates the
effect of food include an increase in subject convenience, thereby
increasing subject compliance, as the subject does not need to
ensure that they are taking a dose either with or without food.
This is significant, as with poor subject compliance an increase in
the medical condition for which the drug is being prescribed may be
observed.
[0048] The difference in absorption of the polycosanol compositions
of the invention, when administered in the fed versus the fasted
state, preferably is less than about 100%, less than about 90%,
less than about 80%, less than about 70%, less than about 60%, less
than about 50%, less than about 40%, less than about 30%, less than
about 25%, less than about 20%, less than about 15%, less than
about 10%, less than about 5%, or less than about 3%.
[0049] D. Dissolution Profiles of the Polycosanol Compositions of
the Invention
[0050] The polycosanol compositions of the invention preferably
have unexpectedly dramatic dissolution profiles. Rapid dissolution
of an administered active agent is preferable, as faster
dissolution generally leads to faster onset of action and greater
bioavailability. To improve the dissolution profile and
bioavailability of polycosanols it would be useful to increase the
drug's dissolution so that it could attain a level close to
100%.
[0051] The polycosanol compositions of the invention preferably
have a dissolution profile in which within about 5 minutes at least
about 20% of the composition is dissolved. In other embodiments of
the invention, at least about 30% or about 40% of the polycosanol
composition is dissolved within about 5 minutes. In yet other
embodiments of the invention, preferably at least about 40%, about
50%, about 60%, about 70%, or about 80% of the polycosanol
composition is dissolved within about 10 minutes. Finally, in
another embodiment of the invention, preferably at least about 70%,
about 80%, about 90%, or about 100% of the polycosanol composition
is dissolved within about 20 minutes.
[0052] Dissolution is preferably measured in a medium which is
discriminating. Such a dissolution medium will produce two very
different dissolution curves for two products having very different
dissolution profiles in gastric juices; i.e., the dissolution
medium is predictive of in vivo dissolution of a composition. An
exemplary dissolution medium is an aqueous medium containing the
surfactant sodium lauryl sulfate at 0.025 M. Determination of the
amount dissolved can be carried out by spectrophotometry. The
rotating blade method (European Pharmacopoeia) can be used to
measure dissolution.
[0053] E. Redispersibility Profiles of the Polycosanol Compositions
of the Invention
[0054] An additional feature of the polycosanol compositions of the
invention is that the compositions preferably redisperse such that
the effective average particle size of the redispersed polycosanol
particles is less than about 2 microns. This is significant, as if
upon administration the nanoparticulate polycosanol compositions of
the invention did not redisperse to a substantially nanoparticulate
particle size, then the dosage form may lose the benefits afforded
by formulating the polycosanol into a nanoparticulate particle
size.
[0055] This is because nanoparticulate active agent compositions
benefit from the small particle size of the active agent; if the
active agent does not redisperse into the small particle sizes upon
administration, then "clumps" or agglomerated active agent
particles are formed, owing to the extremely high surface free
energy of the nanoparticulate system and the thermodynamic driving
force to achieve an overall reduction in free energy. With the
formation of such agglomerated particles, the bioavailability of
the dosage form may fall well below that observed with the liquid
dispersion form of the nanoparticulate active agent.
[0056] Moreover, the nanoparticulate polycosanol compositions of
the invention preferably exhibit dramatic redispersion of the
nanoparticulate polycosanol particles upon administration to a
mammal, such as a human or animal, as demonstrated by
reconstitution/redispersion in a biorelevant aqueous media such
that the effective average particle size of the redispersed
polycosanol particles is less than about 2 microns. Such
biorelevant aqueous media can be any aqueous media that exhibit the
desired ionic strength and pH, which form the basis for the
biorelevance of the media. The desired pH and ionic strength are
those that are representative of physiological conditions found in
the human body. Such biorelevant aqueous media can be, for example,
aqueous electrolyte solutions or aqueous solutions of any salt,
acid, or base, or a combination thereof, which exhibit the desired
pH and ionic strength.
[0057] Biorelevant pH is well known in the art. For example, in the
stomach, the pH ranges from slightly less than 2 (but typically
greater than 1) up to 4 or 5. In the small intestine the pH can
range from 4 to 6, and in the colon it can range from 6 to 8.
Biorelevant ionic strength is also well known in the art. Fasted
state gastric fluid has an ionic strength of about 0.1 M while
fasted state intestinal fluid has an ionic strength of about 0,14.
See e.g., Lindahl et al., "Characterization of Fluids from the
Stomach and Proximal Jejunum in Men and Women," Pharm. Res., 14
(4): 497-502 (1997).
[0058] It is believed that the pH and ionic strength of the test
solution is more critical than the specific chemical content.
Accordingly, appropriate pH and ionic strength values can be
obtained through numerous combinations of strong acids, strong
bases, salts, single or multiple conjugate acid-base pairs (i.e.,
weak acids and corresponding salts of that acid), monoprotic and
polyprotic electrolytes, etc.
[0059] Representative electrolyte solutions can be, but are not
limited to, HCl solutions, ranging in concentration from about
0.001 to about 0.1 M, and NaCl solutions, ranging in concentration
from about 0.001 to about 0.1 M, and mixtures thereof. For example,
electrolyte solutions can be, but are not limited to, about 0.1 M
HCl or less, about 0.01 M HCl or less, about 0.001 M HCl or less,
about 0.1 M NaCl or less, about 0.01 M NaCl or less, about 0.001 M
NaCl or less, and mixtures thereof. Of these electrolyte solutions,
0.01 M HCl and/or 0.1 M NaCl, are most representative of fasted
human physiological conditions, owing to the pH and ionic strength
conditions of the proximal gastrointestinal tract.
[0060] Electrolyte concentrations of 0.001 M HCl, 0.01 M HCl, and
0.1 M HCl correspond to pH 3, pH 2, and pH 1, respectively. Thus, a
0.01 M HCl solution simulates typical acidic conditions found in
the stomach. A solution of 0.1 M NaCl provides a reasonable
approximation of the ionic strength conditions found throughout the
body, including the gastrointestinal fluids, although
concentrations higher than 0.1 M may be employed to simulate fed
conditions within the human GI tract.
[0061] Exemplary solutions of salts, acids, bases or combinations
thereof, which exhibit the desired pH and ionic strength, include
but are not limited to phosphoric acid/phosphate salts+sodium,
potassium and calcium salts of chloride, acetic acid/acetate
salts+sodium, potassium and calcium salts of chloride, carbonic
acid/bicarbonate salts+sodium, potassium and calcium salts of
chloride, and citric acid/citrate salts+sodium, potassium and
calcium salts of chloride.
[0062] In other embodiments of the invention, the redispersed
polycosanol particles of the invention (redispersed in an aqueous,
biorelevant, or any other suitable media) have an effective average
particle size of less than about 1900 nm, less than about 1800 nm,
less than about 1700 nm, less than about 1600 nm, less than about
1500 nm, less than about 1400 nm, less than about 1300 nm, less
than about 1200 nm, less than about 1100 nm, less than about 1000
nm, less than about 900 nm, less than about 800 nm, less than about
700 nm, less than about 600 nm, less than about 500 nm, less than
about 400 nm, less than about 300 nm, less than about 250 nm, less
than about 200 nm, less than about 150 nm, less than about 100 nm,
less than about 75 nm, or less than about 50 nm, as measured by
light-scattering methods, microscopy, or other appropriate
methods.
[0063] By "an effective average particle size of less than about
2000 nm" it is meant that at least 50% of the polycosanol particles
have a particle size of less than the effective average, by weight,
i.e., less than about 2000 nm, 1900 nm, 1800 nm, etc., when
measured by the above-noted techniques. Preferably, at least about
70%, about 90%, about 95%, or about 99% of the polycosanol
particles have a particle size of less than the effective average,
i.e., less than about 2000 nm, 1900 nm, 1800 nm, 1700 nm, etc.
[0064] Redispersibility can be tested using any suitable means
known in the art. See e.g., the example sections of U.S. Pat. No.
6,375,986 for "Solid Dose Nanoparticulate Compositions Comprising a
Synergistic Combination of a Polymeric Surface Stabilizer and
Dioctyl Sodium Sulfosuccinate."
[0065] F. Bioadhesive polycosanol Compositions
[0066] Bioadhesive polycosanol compositions of the invention
comprise at least one cationic surface stabilizer, which are
described in more detail below. Bioadhesive formulations of
polycosanols exhibit exceptional bioadhesion to biological
surfaces, such as mucous. The term bioadhesion refers to any
attractive interaction between two biological surfaces or between a
biological and a synthetic surface. In the case of bioadhesive
nanoparticulate polycosanol compositions, the term bioadhesion is
used to describe the adhesion between the nanoparticulate
polycosanol compositions and a biological substrate (i.e.
gastrointestinal mucin, lung tissue, nasal mucosa, etc.). See e.g.,
U.S. Pat. No. 6,428,814 for "Bioadhesive Nanoparticulate
Compositions Having Cationic Surface Stabilizers," which is
specifically incorporated by reference.
[0067] There are basically two mechanisms which may be responsible
for the bioadhesion phenomena: mechanical or physical interactions
and chemical interactions. The first of these, mechanical or
physical mechanisms, involves the physical interlocking or
interpenetration between a bioadhesive entity and the receptor
tissue, resulting from a good wetting of the bioadhesive surface,
swelling of the bioadhesive polymer, penetration of the bioadhesive
entity into a crevice of the tissue surface, or interpenetration of
bioadhesive composition chains with those of the mucous or other
such related tissues. The second possible mechanism of bioadhesion
incorporates forces such as ionic attraction, dipolar forces, van
der Waals interactions, and hydrogen bonds. It is this form of
bioadhesion which is primarily responsible for the bioadhesive
properties of the nanoparticulate polycosanol compositions of the
invention. However, physical and mechanical interactions may also
play a secondary role in the bioadhesion of such nanoparticulate
compositions.
[0068] The bioadhesive polycosanol compositions of the invention
are useful in any situation in which it is desirable to apply the
compositions to a biological surface. The bioadhesive polycosanol
compositions coat the targeted surface in a continuous and uniform
film which is invisible to the naked human eye.
[0069] A bioadhesive polycosanol composition slows the transit of
the composition, and some polycosanol particles would also most
likely adhere to tissue other than the mucous cells and therefore
give a prolonged exposure to the polycosanol, thereby increasing
absorption and the bioavailability of the administered dosage.
[0070] G. Polycosanol Compositions Used in Conjunction with Other
Active Agents
[0071] The polycosanol compositions of the invention can
additionally comprise one or more non-polycosanol compounds useful:
(1) in treating conditions such as dyslipidemia, hyperlipidemia,
hypercholesterolemia, cardiovascular disorders,
hypertriglyceridemia, coronary heart disease, and peripheral
vascular disease (including symptomatic carotid artery disease), or
related conditions; (2) as adjunctive therapy to diet for the
reduction of LDL-C, total-C, triglycerides, and/or Apo B in adult
patients with primary hypercholesterolemia or mixed dyslipidemia
(Fredrickson Types IIa and IIb); (3) as adjunctive therapy to diet
for treatment of adult patients with hypertriglyceridemia
(Fredrickson Types IV and V hyperlipidemia); (4) in treating
pancreatitis; (5) in treating restenosis; and/or (6) in treating
Alzheimer's disease.
[0072] Exemplary non-polycosanol compositions useful in the
invention include, but are not limited to, cholesterol lowering
agents, alkanoyl L-carnitines, antihypertensives, statins, stanols,
and/or sterols.
[0073] Useful cholesterol lowering agents are well known to those
of skill in the art and include, but are not limited to, ACE
inhibitors, nicotinic acid, niacin, bile acid sequestrants,
fibrates, vitamins, fatty acid derivatives such as fish oil, long
chain plant extract alcohols such as policosinol, ezetimibe, and
celluloses.
[0074] Useful alkanoyl L-carnitines include, but are not limited
to, acetyl L-carnitine, propionyl L-carnitine, butyryl L-carnitine,
valeryl L-carnitine, and isovaleryl L-carnitine, or a
pharmacologically acceptable salt thereof.
[0075] Useful antihypertensives include, but are not limited to
diuretics ("water pills"), beta blockers, alpha blockers,
alpha-beta blockers, sympathetic nerve inhibitors, angiotensin
converting enzyme (ACE) inhibitors, calcium channel blockers,
angiotensin receptor blockers (formal medical name
angiotensin-2-receptor antagonists, known as "sartans" for
short).
[0076] Useful statins include, but are not limited to, atorvastatin
(Lipitor.RTM.) (U.S. Pat. No. 4,681,893) and other
6-[2-(substituted-pyrrol-1-yl)alkyl]pyran-2-ones and derivatives as
disclosed in U.S. Pat. No. 4,647,576); fluvastatin (Lescol.RTM.)
(U.S. Pat. No. 5,354,772); lovastatin (U.S. Pat. No. 4,231,938);
pravastatin (U.S. Pat. No. 4,346,227); simvastatin (U.S. Pat. No.
4,444,784); velostatin; fluindostatin (Sandoz XU-62-320); pyrazole
analogs of mevalonolactone derivatives, as disclosed in PCT
application WO 86/03488; rivastatin and other
pyridyldihydroxyheptenoic acids, as disclosed in European Patent
491226A; Searle's SC-45355 (a 3-substituted pentanedioic acid
derivative); dichloroacetate; imidazole analogs of mevalonolactone,
as disclosed in PCT application WO 86/07054;
3-carboxy-2-hydroxy-propane-phosphonic acid derivatives, as
disclosed in French Patent No. 2,596,393; 2,3-di-substituted
pyrrole, furan, and thiophene derivatives, as disclosed in European
Patent Application No. 0221025; naphthyl analogs of
mevalonolactone, as disclosed in U.S. Pat. No. 4,686,237;
octahydronaphthalenes, such as those disclosed in U.S. Pat. No.
4,499,289; keto analogs of mevinolin (lovastatin), as disclosed in
European Patent Application No. 0,142,146 A2; phosphinic acid
compounds; as well as other HMG CoA reductase inhibitors.
[0077] Useful sterols and/or stanols include, but are not limited
to, plant sterols, plant sterol esters, fish oil, sitosterol,
sitostanol, phytosterol, campestanol, stigmasterol, coprostanol,
cholestanol, and beta-sitosterol.
[0078] The term "stanol" is well known to those skilled in the art
and generally refers to compounds having a saturated
perhydrocyclopentanophenanthrene ring system and having one or more
OH substituents. "Stanols" as used herein mean plant stanol esters,
a food ingredient that can help reduce LDL cholesterol. Plant
stanols are derived from naturally occurring substances in plants
by techniques known to those in the art.
[0079] Such additional compounds can have a conventional
non-nanoparticulate particle size, i.e., an effective average
particle size greater than about 2 microns, or such additional
compounds can be formulated into a nanoparticulate particle size,
i.e., an effective average particle size of less than about 2
microns. If such one or more non-polycosanol compounds have a
nanoparticulate particle size, then preferably such non-polycosanol
compounds are poorly soluble in at least one liquid media (poorly
soluble as defined in the "Definitions" section, above), and have
at least one surface stabilizer adsorbed on or associated with the
surface of the non- polycosanol compound. The one or more surface
stabilizers utilized in the composition of the non- polycosanol
compound can be the same as or different from the one or more
surface stabilizers utilized in the polycosanol composition. A
description of surface stabilizers useful in the invention is
provided below.
II. Compositions
[0080] The present invention is directed to nanoparticulate active
agent compositions comprising at least one polycosanol, and novel
polycosanol combinations. The compositions preferably comprise: (1)
at least one polycosanol or a salt thereof; and (2) at least one
surface stabilizer adsorbed on, or associated with, the surface of
the polycosanol. The nanoparticulate polycosanol particles
preferably have an effective average particle size of less than
about 2000 nm. In another aspect of this invention, novel
combinations of polycosanols and other cholesterol lowering agents
are described and methods of using the same are taught.
[0081] The present invention also includes nanoparticulate
polycosanol compositions together with one or more non-toxic
physiologically acceptable carriers, adjuvants, or vehicles,
collectively referred to as carriers. The compositions can be
formulated for various routes of administration including but not
limited to, oral, rectal, ocular, and parenteral injection (e.g.,
intravenous, intramuscular, or subcutaneous), oral administration
in solid (the preferred route), liquid, or aerosol form, vaginal,
nasal, rectal, ocular, local (e.g., in powder, ointment or drop
form), buccal, intracisternal, intraperitoneal, or topical
administration, and the like.
[0082] A. Polycosanol Particles
[0083] Useful polycosanols include, but are not limited to,
triacontanol, hexacontanol, ecocosanol, hexacosanol, tetracosanol,
dotriacontanol, tetracontanol, or natural products or extracts from
natural products containing such compounds.
[0084] Policosanol (polycosanol) is a complex mixture of
concentrated n-alkyl alcohols derived from, for example, sugar cane
and the wax of honey bees. Polycosanols are extracted by known
methods. These active substances act to lower cholesterol levels by
several mechanisms, including blocking the formation of cholesterol
in the liver.
[0085] As used herein the term "polycosanols" includes polycosanols
or a salt thereof, preferably having a solubility in water of less
than about 30 mg/ml, less than about 20 mg/ml, less than about 10
mg/ml, or more preferably less than about 1 mg/ml.
[0086] The one or more polycosanol particles, or salt thereof, can
be in a crystalline phase, an amorphous phase, a semi-crystalline
phase, a semi-amorphous phase, or a mixture thereof.
[0087] High LDL cholesterol is usually first treated with exercise,
weight loss in obese individuals, and a diet low in cholesterol and
saturated fats. When these measures fail, cholesterol-lowering
medications, such as a polycosanol, can be added. The National
Cholesterol Education Program (NCEP) has published treatment
guidelines for use of polycosanols. These treatment guidelines take
into account the level of LDL cholesterol as well as the presence
of other risk factors such as diabetes, hypertension, cigarette
smoking, low HDL cholesterol level, and family history of early
coronary heart disease.
[0088] B. Surface Stabilizers
[0089] Surface stabilizers especially useful herein physically
adhere on or associate with the surface of the nanoparticulate
polycosanol but do not chemically react with the polycosanol
particles or itself. Preferably, individual molecules of the
surface stabilizer are essentially free of intermolecular
cross-linkages.
[0090] The choice of a surface stabilizer for a polycosanol is
non-trivial and required extensive experimentation to realize a
desirable formulation for the active ingredient's therapeutic
effect desired. For example, the effectiveness of using of a
particular stabilizer with an active ingredient is unpredictable
because the stabilizer among other factors, will affect dissolution
and pharmacokinetic profiles for a polycosanol. Accordingly, the
present invention is directed to the surprising discovery that
stable, therapeutically useful, nanoparticulate polycosanol
compositions can be made.
[0091] Combinations of more than one surface stabilizer can
preferably be used in the invention. Useful surface stabilizers
which can be employed in the invention include, but are not limited
to, known organic and inorganic pharmaceutical excipients. Such
excipients include various polymers, low molecular weight
oligomers, natural products, and surfactants. Preferred surface
stabilizers include nonionic, anionic, cationic, and zwitterionic
surfactants.
[0092] Representative examples of surface stabilizers include
hydroxypropylmethylcellulose (anionic), hydroxypropylcellulose,
polyvinylpyrrolidone, sodium lauryl sulfate, dioctylsulfosuccinate
(anionic), gelatin, casein, lecithin (phosphatides), dextran, gum
acacia, cholesterol, tragacanth, stearic acid, benzalkonium
chloride, calcium stearate, glycerol monostearate, cetostearyl
alcohol, cetomacrogol emulsifying wax, sorbitan esters,
polyoxyethylene alkyl ethers (e.g., macrogol ethers such as
cetomacrogol 1000), polyoxyethylene castor oil derivatives,
polyoxyethylene sorbitan fatty acid esters (e.g., the commercially
available Tweens.RTM. such as e.g., Tween 20.RTM. and Tween 80.RTM.
(ICI Speciality Chemicals)); polyethylene glycols (e.g., Carbowaxs
3550.RTM. and 934.RTM. (Union Carbide)), polyoxyethylene stearates,
colloidal silicon dioxide, phosphates, carboxymethylcellulose
calcium, carboxymethylcellulose sodium, methyl cellulose,
hydroxyethylcellulose, hydroxypropylmethylcellulose phthalate,
noncrystalline cellulose, magnesium aluminium silicate,
triethanolamine, polyvinyl alcohol (PVA),
4-(1,1,3,3-tetramethylbutyl)-phenol polymer with ethylene oxide and
formaldehyde (also known as tyloxapol, superione, and triton),
poloxamers (e.g., Pluronics F68.RTM. and F108.RTM., which are block
copolymers of ethylene oxide and propylene oxide); poloxamines
(e.g., Tetronic 908.RTM., also known as Poloxamine 908.RTM., which
is a tetrafunctional block copolymer derived from sequential
addition of propylene oxide and ethylene oxide to ethylenediamine
(BASF Wyandotte Corporation, Parsippany, N.J.)); Tetronic 1508.RTM.
(T-1508) (BASF Wyandotte Corporation), Triton X-200.RTM., which is
an alkyl aryl polyether sulfonate (Dow Chemical); Crodestas
F-110.RTM., which is a mixture of sucrose stearate and sucrose
distearate (Croda Inc.); p-isononylphenoxypoly-(glycidol), also
known as Olin-10G.RTM. or Surfactant 10-G.RTM. (Olin Chemicals,
Stamford, CT); Crodestas SL-40.RTM. (Croda, Inc.); and SA9OHCO,
which is
C.sub.18H.sub.37CH.sub.2(CON(CH.sub.3)--CH.sub.2(CHOH).sub.4(CH.sub.2OH).-
sub.2 (Eastman Kodak Co.); decanoyl-N-methylglucamide; n-decyl
.beta.-D-glucopyranoside; n-decyl .beta.-D-maltopyranoside;
n-dodecyl .beta.-D-glucopyranoside; n-dodecyl .beta.-D-maltoside;
heptanoyl-N-methylglucamide; n-heptyl-.beta.-D-glucopyranoside;
n-heptyl .beta.-D-thioglucoside; n-hexy .beta.-D-glucopyranoside;
nonanoyl-N-methylglucamide; n-noyl .beta.-D-glucopyranoside;
octanoyl-N-methylglucamide; n-octyl-.beta.-D-glucopyranoside; octyl
.beta.-D-thioglucopyranoside; PEG-derivatized phospholipid, PEG-
derivatized cholesterol, PEG-derivatized cholesterol derivative,
PEG- derivatized vitamin A, PEG- derivatized vitamin E, lysozyme,
random copolymers of vinyl pyrrolidone and vinyl acetate, and the
like such as Plasdone.RTM. S630 in a 60:40 ratio of the pyrrolidone
and vinyl acetate.
[0093] More examples of useful surface stabilizers include, but are
not limited to, polymers, biopolymers, polysaccharides,
cellulosics, alginates, phospholipids, and nonpolymeric compounds,
such as zwitterionic stabilizers, poly-n-methylpyridinium, anthryul
pyridinium chloride, cationic phospholipids, chitosan, polylysine,
polyvinylimidazole, polybrene, polymethylmethacrylate
trimethylammoniumbromide bromide (PMMTMABr),
hexadecyltrimethylammonium bromide (HDMAB), and
polyvinylpyrrolidone-2-dimethylaminoethyl methacrylate dimethyl
sulfate.
[0094] Other useful cationic stabilizers include, but are not
limited to, cationic lipids, sulfonium, phosphonium, and
quarternary ammonium compounds, such as stearyltrimethylammonium
chloride, benzyl-di(2-chloroethyl)ethylammonium bromide, coconut
trimethyl ammonium chloride or bromide, coconut methyl
dihydroxyethyl ammonium chloride or bromide, decyl triethyl
ammonium chloride, decyl dimethyl hydroxyethyl ammonium chloride or
bromide, C.sub.12-15dimethyl hydroxyethyl ammonium chloride or
bromide, coconut dimethyl hydroxyethyl ammonium chloride or
bromide, myristyl trimethyl ammonium methyl sulphate, lauryl
dimethyl benzyl ammonium chloride or bromide, lauryl dimethyl
(ethenoxy).sub.4 ammonium chloride or bromide, N-alkyl
(C.sub.12-18)dimethylbenzyl ammonium chloride, N-alkyl
(C.sub.14-18)dimethyl-benzyl ammonium chloride,
N-tetradecylidmethylbenzyl ammonium chloride monohydrate, dimethyl
didecyl ammonium chloride, N-alkyl and (C.sub.12-14) dimethyl
1-napthylmethyl ammonium chloride, trimethylammonium halide,
alkyl-trimethylammonium salts and dialkyl-dimethylammonium salts,
lauryl trimethyl ammonium chloride, ethoxylated
alkyamidoalkyldialkylammonium salt and/or an ethoxylated trialkyl
ammonium salt, dialkylbenzene dialkylammonium chloride,
N-didecyldimethyl ammonium chloride, N-tetradecyldimethylbenzyl
ammonium, chloride monohydrate, N-alkyl(C.sub.12-14) dimethyl
1-naphthylmethyl ammonium chloride and dodecyldimethylbenzyl
ammonium chloride, dialkyl benzenealkyl ammonium chloride, lauryl
trimethyl ammonium chloride, alkylbenzyl methyl ammonium chloride,
alkyl benzyl dimethyl ammonium bromide, C.sub.12, C.sub.15,
C.sub.17 trimethyl ammonium bromides, dodecylbenzyl triethyl
ammonium chloride, poly-diallyldimethylammonium chloride (DADMAC),
dimethyl ammonium chlorides, alkyldimethylammonium halogenides,
tricetyl methyl ammonium chloride, decyltrimethylammonium bromide,
dodecyltriethylammonium bromide, tetradecyltrimethylammonium
bromide, methyl trioctylammonium chloride (ALIQUAT 336.TM.),
POLYQUAT 10.TM., tetrabutylammonium bromide, benzyl
trimethylammonium bromide, choline esters (such as choline esters
of fatty acids), benzalkonium chloride, stearalkonium chloride
compounds (such as stearyltrimonium chloride and Di-stearyldimonium
chloride), cetyl pyridinium bromide or chloride, halide salts of
quaternized polyoxyethylalkylamines, MIRAPOL.TM. and ALKAQUAT.TM.
(Alkaril Chemical Company), alkyl pyridinium salts; amines, such as
alkylamines, dialkylamines, alkanolamines, polyethylenepolyamines,
N,N-dialkylaminoalkyl acrylates, and vinyl pyridine, amine salts,
such as lauryl amine acetate, stearyl amine acetate,
alkylpyridinium salt, and alkylimidazolium salt, and amine oxides;
imide azolinium salts; protonated quaternary acrylamides;
methylated quaternary polymers, such as poly[diallyl
dimethylammonium chloride] and poly-[N-methyl vinyl pyridinium
chloride]; and cationic guar.
[0095] Such exemplary cationic surface stabilizers and other useful
cationic surface stabilizers are described in J. Cross and E.
Singer, Cationic Surfactants: Analytical and Biological Evaluation
(Marcel Dekker, 1994); P. and D. Rubingh (Editor), Cationic
Surfactants: Physical Chemistry (Marcel Dekker, 1991); and J.
Richmond, Cationic Surfactants: Organic Chemistry, (Marcel Dekker,
1990).
[0096] Nonpolymeric surface stabilizers are any nonpolymeric
compound, such benzalkonium chloride, a carbonium compound, a
phosphonium compound, an oxonium compound, a halonium compound, a
cationic organometallic compound, a quarternary phosphorous
compound, a pyridinium compound, an anilinium compound, an ammonium
compound, a hydroxylammonium compound, a primary ammonium compound,
a secondary ammonium compound, a tertiary ammonium compound, and
quarternary ammonium compounds of the formula
NR.sub.1R.sub.2R.sub.3R.sub.4.sup.(+). For compounds of the formula
NR.sub.1R.sub.2R.sub.3R.sub.4.sup.(+): [0097] none of
R.sub.1-R.sub.4 are CH.sub.3; [0098] (ii) one of R.sub.1-R.sub.4 is
CH.sub.3; [0099] (iii) three of R.sub.1-R.sub.4 are CH.sub.3;
[0100] (iv) all of R.sub.1-R.sub.4 are CH.sub.3; [0101] (v) two of
R.sub.1-R.sub.4 are CH.sub.3, one of R.sub.1-R.sub.4 is
C.sub.6H.sub.5CH.sub.2, and one of R.sub.1-R.sub.4 is alkyl chain
of seven carbon atoms or less; [0102] (vi) two of R.sub.1-R.sub.4
are CH.sub.3, one of R.sub.1-R.sub.4 is C.sub.6H.sub.5CH.sub.2, and
one of R.sub.1-R.sub.4 is alkyl chain of nineteen carbon atoms or
more; [0103] (vii) two of R.sub.1-R.sub.4 are CH.sub.3 and one of
R.sub.1-R.sub.4 is the group C.sub.6H.sub.5(CH.sub.2).sub.n, where
n>1; [0104] (viii) two of R.sub.1-R.sub.4 are CH.sub.3, one of
R.sub.1-R.sub.4 is C.sub.6H.sub.5CH.sub.2, and one of
R.sub.1-R.sub.4 comprises at least one heteroatom; [0105] (ix) two
of R.sub.1-R.sub.4 are CH.sub.3, one of R.sub.1-R.sub.4 is
C.sub.6H.sub.5CH.sub.2, and one of R.sub.1-R.sub.4 comprises at
least one halogen; [0106] (x) two of R.sub.1-R.sub.4 are CH.sub.3,
one of R.sub.1-R.sub.4 is C.sub.6H.sub.5CH.sub.2, and one of
R.sub.1-R.sub.4 comprises at least one cyclic fragment; [0107] (xi)
two of R.sub.1-R.sub.4 are CH.sub.3 and one of R.sub.1-R.sub.4 is a
phenyl ring; or [0108] (xii) two of R.sub.1-R.sub.4 are CH.sub.3
and two of R.sub.1-R.sub.4 are purely aliphatic fragments.
[0109] Such compounds include, but are not limited to,
behenalkonium chloride, benzethonium chloride, cetylpyridinium
chloride, behentrimonium chloride, lauralkonium chloride,
cetalkonium chloride, cetrimonium bromide, cetrimonium chloride,
cethylamine hydrofluoride, chlorallylmethenamine chloride
(Quaternium-15), distearyldimonium chloride (Quaternium-5), dodecyl
dimethyl ethylbenzyl ammonium chloride (Quaternium-14),
Quaternium-22, Quaternium-26, Quaternium-18 hectorite,
dimethylaminoethylchloride hydrochloride, cysteine hydrochloride,
diethanolammonium POE (10) oletyl ether phosphate,
diethanolammonium POE (3)oleyl ether phosphate, tallow alkonium
chloride, dimethyl dioctadecylammoniumbentonite, stearalkonium
chloride, domiphen bromide, denatonium benzoate, myristalkonium
chloride, laurtrimonium chloride, ethylenediamine dihydrochloride,
guanidine hydrochloride, pyridoxine HCl, iofetamine hydrochloride,
meglumine hydrochloride, methylbenzethonium chloride, myrtrimonium
bromide, oleyltrimonium chloride, polyquaternium-1,
procainehydrochloride, cocobetaine, stearalkonium bentonite,
stearalkoniumhectonite, stearyl trihydroxyethyl propylenediamine
dihydrofluoride, tallowtrimonium chloride, and hexadecyltrimethyl
ammonium bromide.
[0110] Most of these surface stabilizers are known pharmaceutical
excipients and are described in detail in the Handbook of
Pharmaceutical Excipients, published jointly by the American
Pharmaceutical Association and The Pharmaceutical Society of Great
Britain (The Pharmaceutical Press, 2000), specifically incorporated
by reference.
[0111] The surface stabilizers are commercially available and/or
can be prepared by techniques known in the art.
[0112] C. Other Pharmaceutical Excipients
[0113] Pharmaceutical compositions according to the invention may
also comprise one or more binding agents, filling agents,
lubricating agents, suspending agents, sweeteners, flavoring
agents, preservatives, buffers, wetting agents, disintegrants,
effervescent agents, and other excipients depending upon the route
of administration and the dosage form desired. Such excipients are
known in the art.
[0114] Examples of filling agents are lactose monohydrate, lactose
anhydrous, and various starches; examples of binding agents are
various celluloses and cross-linked polyvinylpyrrolidone,
microcrystalline cellulose, such as Avicel.RTM. PH101 and
Avicel.RTM. PH102, microcrystalline cellulose, and silicified
microcrystalline cellulose (ProSolv SMCC.TM.).
[0115] Suitable lubricants, including agents that act on the
flowability of the powder to be compressed, are colloidal silicon
dioxide, such as Aerosil.RTM. 200, talc, stearic acid, magnesium
stearate, calcium stearate, and silica gel.
[0116] Examples of sweeteners are any natural or artificial
sweetener, such as sucrose, xylitol, sodium saccharin, cyclamate,
aspartame, and acsulfame. Examples of flavoring agents are
Magnasweet.RTM. (trademark of MAFCO), bubble gum flavor, and fruit
flavors, and the like.
[0117] Examples of preservatives are potassium sorbate,
methylparaben, propylparaben, benzoic acid and its salts, other
esters of parahydroxybenzoic acid such as butylparaben, alcohols
such as ethyl or benzyl alcohol, phenolic compounds such as phenol,
or quarternary compounds such as benzalkonium chloride.
[0118] Suitable diluents include pharmaceutically acceptable inert
fillers, such as microcrystalline cellulose, lactose, dibasic
calcium phosphate, saccharides, and/or mixtures of any of the
foregoing. Examples of diluents include microcrystalline cellulose,
such as Avicel.RTM. PH101 and Avicel.RTM. PH102; lactose such as
lactose monohydrate, lactose anhydrous, and Pharmatose.RTM. DCL21;
dibasic calcium phosphate such as Emcompress.RTM.; mannitol;
starch; sorbitol; sucrose; and glucose.
[0119] Suitable disintegrants include lightly crosslinked polyvinyl
pyrrolidone, corn starch, potato starch, maize starch, and modified
starches, croscarmellose sodium, cross-povidone, sodium starch
glycolate, and mixtures thereof.
[0120] Examples of effervescent agents are effervescent couples
such as an organic acid and a carbonate or bicarbonate. Suitable
organic acids include, for example, citric, tartaric, malic,
fumaric, adipic, succinic, and alginic acids and anhydrides and
acid salts. Suitable carbonates and bicarbonates include, for
example, sodium carbonate, sodium bicarbonate, potassium carbonate,
potassium bicarbonate, magnesium carbonate, sodium glycine
carbonate, L-lysine carbonate, and arginine carbonate.
Alternatively, only the sodium bicarbonate component of the
effervescent couple may be present.
[0121] D. Nanoparticulate Polycosanol Particle Size
[0122] The compositions of the invention contain polycosanol
nanoparticles, such as sitosterol and/or phytosterol nanoparticles,
which have an effective average particle size of less than about
2000 nm (i.e., 2 microns). In a preferred embodiment of the
invention, the polycosanol nanoparticles have an effective average
particle size of less than about 1900 nm, less than about 1800 nm,
less than about 1700 nm, less than about 1600 nm, less than about
1500 nm, less than about 1400 nm, less than about 1300 nm, less
than about 1200 nm, less than about 1100 nm, less than about 1000
nm, less than about 900 nm, less than about 800 nm, less than about
700 nm, less than about 600 nm, less than about 500 nm, less than
about 400 nm, less than about 300 nm, less than about 250 nm, less
than about 200 nm, less than about 150 nm, less than about 100 nm,
less than about 75 nm, or less than about 50 nm, as measured by
light-scattering methods, microscopy, or other appropriate
methods.
[0123] By "an effective average particle size of less than about
2000 nm" it is meant that at least 50% of the polycosanol particles
have a particle size less than the effective average, by weight,
i.e., less than about 2000 nm, about 1900 nm, about 1800 nm, etc.,
when measured by the above-noted techniques. Preferably, at least
about 70%, about 90%, about 95%, or about 99% of the polycosanol
particles have a particle size less than the effective average,
i.e., less than about 2000 nm, about 1900 nm, about 1800 nm,
etc.
[0124] In the present invention, the value for D50 of a
nanoparticulate polycosanol composition is the particle size below
which 50% of the polycosanol particles fall, by weight. Similarly,
D90 is the particle size below which 90% of the polycosanol/stanol
particles fall, by weight.
[0125] E. Concentration of Nanoparticulate Polycosanol and Surface
Stabilizers
[0126] The relative amounts of at least one polycosanol and one or
more surface stabilizers can vary widely. The optimal amount of the
individual components depends, for example, upon one or more of the
physical and chemical attributes of the particular polycosanol
selected and surface stabilizer(s) selected, such as the
hydrophilic lipophilic balance (HLB), melting point, and the
surface tension of water solutions of the stabilizer, etc.
[0127] Preferably, the concentration of the at least one
polycosanol can vary from about 99.5% to about 0.001%, preferably
from about 95% to about 0.1%, preferably from about 90% to about
0.5%, by weight, based on the total combined weight of the
polycosanol and at least one surface stabilizer, not including
other excipients. Higher concentrations of the active ingredient
are generally preferred from a dose and cost efficiency standpoint.
Preferably, the concentration of the at least one surface
stabilizer can vary from about 0.5% to about 99.999%, from about
5.0% to about 99.9%, or from about 10% to about 99.5%, by weight,
based on the total combined dry weight of the polycosanol and at
least one surface stabilizer, not including other excipients.
[0128] Exemplary useful ratios of active ingredient to stabilizers
herein are preferably about 1:1, preferably about 2:1, preferably
about 3:1, preferably about 4:1, preferably about 5:1, preferably
about 6:1, preferably about 7:1, preferably about 8:1, and
preferably about 10:1, by weight, based on the total combined dry
weight of the polycosanol and at least one surface stabilizer, not
including other excipients.
[0129] III. Methods of Making Nanoparticulate Polycosanol
Compositions
[0130] The nanoparticulate polycosanol compositions can be made
using any suitable method known in the art such as, for example,
milling, homogenization, or precipitation techniques. Exemplary
methods of making nanoparticulate compositions are described in the
'684 patent. Methods of making nanoparticulate compositions are
also described in U.S. Pat. No. 5,518,187 for "Method of Grinding
Pharmaceutical Substances;" U.S. Pat. No. 5,718,388 for "Continuous
Method of Grinding Pharmaceutical Substances;" U.S. Pat. No.
5,862,999 for "Method of Grinding Pharmaceutical Substances;" U.S.
Pat. No. 5,665,331 for "Co-Microprecipitation of Nanoparticulate
Pharmaceutical Agents with Crystal Growth Modifiers;" U.S. Pat. No.
5,662,883 for "Co-Microprecipitation of Nanoparticulate
Pharmaceutical Agents with Crystal Growth Modifiers;" U.S. Pat. No.
5,560,932 for "Microprecipitation of Nanoparticulate Pharmaceutical
Agents;" U.S. Pat. No. 5,543,133 for "Process of Preparing X-Ray
Contrast Compositions Containing Nanoparticles;" U.S. Pat. No.
5,534,270 for "Method of Preparing Stable Drug Nanoparticles;" U.S.
Pat. No. 5,510,118 for "Process of Preparing Therapeutic
Compositions Containing Nanoparticles;" and U.S. Pat. No. 5,470,583
for "Method of Preparing Nanoparticle Compositions Containing
Charged Phospholipids to Reduce Aggregation," all of which are
specifically incorporated by reference.
[0131] The resultant nanoparticulate polycosanol compositions or
dispersions can be utilized in solid or liquid dosage formulations,
such as liquid dispersions, gels, aerosols, ointments, creams,
controlled release formulations, fast melt formulations,
lyophilized formulations, tablets, capsules, delayed release
formulations, extended release formulations, pulsatile release
formulations, mixed immediate release and controlled release
formulations, etc. Solid dose forms of the dispersions of novel
polycosanol formulations according to the present invention can be
made as described in U.S. Pat. No. 6,375,986.
[0132] A. Milling to Obtain Nanoparticulate Polycosanol
Dispersions
[0133] Milling a polycosanol to obtain a nanoparticulate
polycosanol dispersion comprises dispersing polycosanol particles
in a liquid dispersion medium in which the polycosanol is poorly
soluble, followed by applying mechanical means in the presence of
grinding media to reduce the particle size of the polycosanol to
the desired effective average particle size. The dispersion medium
can be, for example, water, safflower oil, ethanol, t-butanol,
glycerin, polyethylene glycol (PEG), hexane, or glycol.
[0134] The polycosanol particles can be reduced in size preferably
in the presence of at least one surface stabilizer. Alternatively,
the polycosanol particles can be contacted with one or more surface
stabilizers after attrition. Other compounds, such as a diluent,
can be added to the polycosanol/surface stabilizer composition
during the size reduction process. Dispersions can be manufactured
continuously or in a batch mode.
[0135] B. Precipitation to Obtain Nanoparticulate Polycosanol
Compositions
[0136] Another method of forming the desired nanoparticulate
polycosanol composition is by microprecipitation. This is a method
of preparing stable dispersions of poorly soluble active agents in
the presence of one or more surface stabilizers and one or more
colloid stability enhancing surface active agents free of any trace
toxic solvents or solubilized heavy metal impurities. Such a method
comprises, for example: (1) dissolving a polycosanol in a suitable
solvent; (2) adding the formulation from step (1) to a solution
comprising at least one surface stabilizer; and (3) precipitating
the formulation from step (2) using an appropriate non-solvent. The
method can be followed by removal of any formed salt, if present,
by dialysis or diafiltration and concentration of the dispersion by
conventional means.
[0137] C. Homogenization to Obtain Polycosanol Nanoparticulate
Compositions
[0138] Exemplary homogenization methods of preparing active agent
nanoparticulate compositions are described in U.S. Pat. No.
5,510,118, for "Process of Preparing Therapeutic Compositions
Containing Nanoparticles." Such a method comprises dispersing
polycosanol particles in a liquid dispersion medium in which the
polycosanol is poorly soluble, followed by subjecting the
dispersion to homogenization to reduce the particle size of the
polycosanol to the desired effective average particle size. The
polycosanol particles are preferably reduced in size in the
presence of at least one surface stabilizer. Alternatively, the
polycosanol particles can be contacted with one or more surface
stabilizers either before or after attrition. Other compounds, such
as a diluent, can be added to the polycosanol/surface stabilizer
composition before, during, or after the size reduction process.
Dispersions can be manufactured continuously or in a batch
mode.
IV. Methods of Using Polycosanol Compositions of the Current
Invention
[0139] The polycosanol compositions of the present invention can be
administered to a subject via any conventional means including, but
not limited to, preferably orally, rectally, ocularly, parenterally
(e.g., intravenous, intramuscular, or subcutaneous),
intracisternally, pulmonary, intravaginally, intraperitoneally,
locally (e.g., powders, ointments or drops), or as a buccal or
nasal spray. As used herein, the term "subject" is used to mean an
animal, preferably a mammal, including a human or non-human. The
terms patient and subject may be used interchangeably.
[0140] The present invention provides a method of prolonging plasma
levels of a polycosanol in a subject while achieving the desired
therapeutic effect. In one aspect, such a method comprises orally
administering to a subject an effective amount of a composition of
this invention comprising a polycosanol.
[0141] In one aspect, the compositions of the invention are useful
in treating conditions that may be directly or indirectly
associated with elevated and/or uncontrolled cholesterol metabolism
as described herein and known to those in the art.
[0142] Compositions suitable for parenteral injection may comprise
physiologically acceptable sterile aqueous or nonaqueous solutions,
dispersions, suspensions or emulsions, and sterile powders for
reconstitution into sterile injectable solutions or dispersions.
Examples of suitable aqueous and nonaqueous carriers, diluents,
solvents, or vehicles include water, ethanol, polyols
(propyleneglycol, polyethylene-glycol, glycerol, and the like),
suitable mixtures thereof, vegetable oils (such as olive oil) and
injectable organic esters such as ethyl oleate. Proper fluidity can
be maintained, for example, by the use of a coating such as
lecithin, by the maintenance of the required particle size in the
case of dispersions, and by the use of surfactants.
[0143] The nanoparticulate polycosanol compositions may also
contain adjuvants such as preserving, wetting, emulsifying, and
dispensing agents. Prevention of the growth of microorganisms can
also be ensured by various antibacterial and antifungal agents,
such as parabens, chlorobutanol, phenol, sorbic acid, and the like.
It may also be desirable to include isotonic agents, such as
sugars, sodium chloride, and the like. Prolonged absorption of the
injectable pharmaceutical form can be brought about by the use of
agents delaying absorption, such as aluminum monostearate and
gelatin.
[0144] Solid dosage forms for oral administration are preferred and
include, but are not limited to, capsules, tablets, pills, powders,
caplets, and granules. In such solid dosage forms, the active agent
(i.e., the composition of this invention) is admixed with at least
one of the following: (a) one or more inert excipients (or
carriers), such as sodium citrate or dicalcium phosphate; (b)
fillers or extenders, such as starches, lactose, sucrose, glucose,
mannitol, and silicic acid; (c) binders, such as
carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone,
sucrose, and acacia; (d) humectants, such as glycerol; (e)
disintegrating agents, such as agar-agar, calcium carbonate, potato
or tapioca starch, alginic acid, certain complex silicates, and
sodium carbonate; (f) solution retarders, such as paraffin; (g)
absorption accelerators, such as quaternary ammonium compounds; (h)
wetting agents, such as cetyl alcohol and glycerol monostearate;
(i) adsorbents, such as kaolin and bentonite; and (j) lubricants,
such as talc, calcium stearate, magnesium stearate, solid
polyethylene glycols, sodium lauryl sulfate, or mixtures thereof.
For capsules, tablets, and pills, the dosage forms may also
comprise buffering agents.
[0145] Liquid dosage forms for oral administration include
pharmaceutically acceptable dispersions, emulsions, solutions,
suspensions, syrups, and elixirs. In addition to the active agent,
the liquid dosage forms may comprise inert diluents commonly used
in the art, such as water or other solvents, solubilizing agents,
and emulsifiers. Exemplary emulsifiers are ethyl alcohol, isopropyl
alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl
benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide,
oils, such as cottonseed oil, groundnut oil, corn germ oil, olive
oil, castor oil, and sesame oil, glycerol, tetrahydrofurfuryl
alcohol, polyethyleneglycols, fatty acid esters of sorbitan, or
mixtures of these substances, and the like.
[0146] Besides such inert diluents, the composition can also
include adjuvants, such as wetting agents, emulsifying and
suspending agents, sweetening, flavoring, and perfuming agents.
[0147] The effective amounts of the polycosanol compositions of the
invention can be determined empirically and can be employed in pure
form or, where such forms exist, in pharmaceutically acceptable
salt, ester, or prodrug form. Actual dosage levels of polycosanol
in the nanoparticulate compositions of the invention may be varied
to obtain an amount of polycosanol that is effective to obtain a
desired therapeutic response for a particular composition, method
of administration, and the condition to be treated. The selected
dosage level therefore depends upon the desired therapeutic effect,
the route of administration, the potency of the administered
polycosanol, the desired duration of treatment, and other
factors.
[0148] Dosage unit compositions may contain amounts of submultiples
thereof as may be used to make up the daily dose. It will be
understood, however, that the specific dose level for any
particular patient will depend upon a variety of factors: the type
and degree of the cellular or physiological response to be
achieved; activity of the specific agent or composition employed;
the specific agents or composition employed; the age, body weight,
general health, sex, and diet of the patient; the time of
administration, route of administration and rate of excretion of
the agent; the duration of the treatment; drugs used in combination
or coincidental with the specific agent; and like factors well
known in the medical arts.
V. Polycosanol Combinations
[0149] Polycosanol compositions of the present invention are also
particularly useful when given pursuant to the method of this
invention in combination with a therapeutically effective amount of
at least one other non-polycosanol active agent useful: (1) in
treating conditions such as dyslipidemia, hyperlipidemia,
hypercholesterolemia, cardiovascular disorders,
hypertriglyceridemia, coronary heart disease, and peripheral
vascular disease (including symptomatic carotid artery disease), or
related conditions; (2) as adjunctive therapy to diet for the
reduction of LDL-C, total-C, triglycerides, and/or Apo B in adult
patients with primary hypercholesterolemia or mixed dyslipidemia
(Fredrickson Types IIa and IIb); (3) as adjunctive therapy to diet
for treatment of adult patients with hypertriglyceridemia
(Fredrickson Types IV and V hyperlipidemia); (4) in treating
pancreatitis; (5) in treating restenosis; and/or (6) in treating
Alzheimer's disease.
[0150] Exemplary non-polycosanol compositions useful in the
invention include, e.g., cholesterol lowering agents, alkanoyl
L-carnitines, antihypertensives, statins, sterols, and/or
stanols.
[0151] Useful cholesterol lowering agents are well known to those
of skill in the art and include, but are not limited to, ACE
inhibitors, nicotinic acid, niacin, bile acid sequestrants,
fibrates, vitamins, fatty acid derivatives such as fish oil, long
chain plant extract alcohols such as policosinol, ezetimibe, and
celluloses.
[0152] Useful alkanoyl L-carnitines include, but are not limited
to, acetyl L-carnitine, propionyl L-carnitine, butyryl L-carnitine,
valeryl L-carnitine, and isovaleryl L-carnitine, or a
pharmacologically acceptable salt thereof
[0153] Useful antihypertensives include, but are not limited to
diuretics ("water pills"), beta blockers, alpha blockers,
alpha-beta blockers, sympathetic nerve inhibitors, angiotensin
converting enzyme (ACE) inhibitors, calcium channel blockers,
angiotensin receptor blockers (formal medical name
angiotensin-2-receptor antagonists, known as "sartans" for
short).
[0154] Useful statins include, but are not limited to, atorvastatin
(Lipitor.RTM.) (U.S. Pat. No. 4,681,893) and other
6-[2-(substituted-pyrrol-1-yl)alkyl]pyran-2-ones and derivatives as
disclosed in U.S. Pat. No. 4,647,576); fluvastatin (Lescol.RTM.)
(U.S. Pat. No. 5,354,772); lovastatin (U.S. Pat. No. 4,231,938);
pravastatin (U.S. Pat. No. 4,346,227); simvastatin (U.S. Pat. No.
4,444,784); velostatin; fluindostatin (Sandoz XU-62-320); pyrazole
analogs of mevalonolactone derivatives, as disclosed in PCT
application WO 86/03488; rivastatin and other
pyridyldihydroxyheptenoic acids, as disclosed in European Patent
491226A; Searle's SC-45355 (a 3-substituted pentanedioic acid
derivative); dichloroacetate; imidazole analogs of mevalonolactone,
as disclosed in PCT application WO 86/07054;
3-carboxy-2-hydroxy-propane-phosphonic acid derivatives, as
disclosed in French Patent No. 2,596,393; 2,3-di-substituted
pyrrole, furan, and thiophene derivatives, as disclosed in European
Patent Application No. 0221025; naphthyl analogs of
mevalonolactone, as disclosed in U.S. Pat. No. 4,686,237;
octahydronaphthalenes, such as those disclosed in U.S. Pat. No.
4,499,289; keto analogs of mevinolin (lovastatin), as disclosed in
European Patent Application No. 0,142,146 A2; phosphinic acid
compounds; as well as other HMG CoA reductase inhibitors.
[0155] Useful sterols and/or stanols include, but are not limited
to, plant sterols, plant sterol esters, fish oil, sitosterol,
sitostanol, phytosterol, campestanol, stigmasterol, coprostanol,
cholestanol, and beta-sitosterol.
[0156] The term "stanol" is well known to those skilled in the art
and generally refers to compounds having a saturated
perhydrocyclopentanophenanthrene ring system and having one or more
OH substituents. "Stanols" as used herein mean plant stanol esters,
a food ingredient that can help reduce LDL cholesterol. Plant
stanols are derived from naturally occurring substances in plants
by techniques known to those in the art.
[0157] The following examples are given to illustrate the present
invention. It should be understood, however, that the invention is
not to be limited to the specific conditions or details described
in these examples. Throughout the specification, any and all
references to a publicly available document, including a U.S.
patent, are specifically incorporated by reference.
[0158] In the examples that follow, the particle sizes were
measured using a Horiba LA-910 Laser Scattering Particle Size
Distribution Analyzer (Horiba Instruments, Irvine, Calif.). The
particle mean and D.sub.90 (which is the size below which 90% of
the distribution is located) are obtained from a weight
distribution. All formulations are given in weight % (w/w).
Example 1
[0159] The purpose of this example was to identify formulations
that would produce stable nanoparticulate dispersions of
polycosanol.
[0160] Arriving at a formulation that results in a stable
dispersion having a small particle size is nontrivial and requires
extensive experimentation.
[0161] Two grades of polycosanol were evaluated, labeled OCTA-60
(Formulation A) and OCTA-95 (Formulation B). The 1-octacosanol
content is ca 60% in Formulation A and ca 95% in Formulation B.
Both contain a total of ca 97 to 98% long chain aliphatic alcohols,
such as 1-octacosanol, 1-triacontanol, 1-dotriactontanol,
1-hexacosanol, and 1-heptacosanol.
[0162] The polycosanol was commercially obtained from Garuda
International, Lemon Cove, Calif. and the specifications for each
product used herein are available from the company's web site at
http://www.garudaint.com/.
[0163] Formulation A, comprising 5% (w/w) polycosanol OCTA-60 and
1% (w/w) Tween.RTM. 80, and Formulation B, comprising 5% (w/w)
polycosanol OCTA-95 and 1% (w/w) Tween.RTM. 80, were each processed
in a DYNO.RTM.-Mill KDL (Willy A. Bachofen A G, Maschinenfabrik,
Basel, Switzerland) equipped with a 150 cc batch chamber using a
500 .mu.m milling media of type Polymill.RTM. 500 for 6.5-7 hrs at
10.degree. C.
[0164] The polycosanol particle sizes for Formulations A and B were
measured using a Horiba LA-910 Laser Scattering Particle Size
Distribution Analyzer (Horiba Instruments, Irvine, Calif.). The
polycosanol particle mean and D.sub.90 were obtained from a weight
distribution. The results are shown in Table 2, below.
TABLE-US-00001 TABLE 1 Size after 2 Size at harvest weeks at
25.degree. C. 1 min Active ingredient Stabilizer Mean (nm) D90 (nm)
Mean (nm) D90 (nm) sonication A. 5% Polycosanol 1.0% Tween.sup.
.RTM. 80 292,000 579,000 710 998 N OCTA-60 (Polysorbate 80) 317 496
317 998 Y B. 5% Polycosanol 1.0% Tween.sup. .RTM. 80 240 368 245
385 N OCTA-95 (Polysorbate 80) 236 362 221 342 Y
[0165] The results show that the product of higher purity, OCTA-95,
produces a more stable dispersion as indicated by the size before
and after sonication. It should be noted, however, that while the
OCTA-60 formulation initially seems prone to aggregation, it
relaxes into a more stable dispersion upon aging. Thus, both types
of polycosanol are suitable for the nanoparticulate polycosanol
compositions of the invention.
[0166] It will be apparent to those skilled in the art that various
modifications and variations can be made in the methods and
compositions of the present invention without departing from the
spirit or scope of the invention. Thus, it is intended that the
present invention cover the modifications and variations of this
invention provided they come within the scope of the appended
claims and their equivalents.
* * * * *
References