U.S. patent application number 15/478472 was filed with the patent office on 2017-07-20 for stable compositions of peptide epoxy ketones.
This patent application is currently assigned to InnoPharma, Inc.. The applicant listed for this patent is InnoPharma, Inc.. Invention is credited to Ketan Hippalgaonkar, Satish Pejaver, Navneet Puri, Kumaresh Soppimath.
Application Number | 20170202901 15/478472 |
Document ID | / |
Family ID | 50233867 |
Filed Date | 2017-07-20 |
United States Patent
Application |
20170202901 |
Kind Code |
A1 |
Hippalgaonkar; Ketan ; et
al. |
July 20, 2017 |
STABLE COMPOSITIONS OF PEPTIDE EPOXY KETONES
Abstract
The invention relates to pharmaceutical compositions that
provide improved solubility and stability for peptide epoxy
ketones. More specifically, the invention relates to pharmaceutical
compositions comprising the peptide epoxy ketone proteasome
inhibitor carfilzomib.
Inventors: |
Hippalgaonkar; Ketan;
(Highland Park, NJ) ; Soppimath; Kumaresh;
(Monmouth, NJ) ; Pejaver; Satish; (Bridgewater,
NJ) ; Puri; Navneet; (Bridgewater, NJ) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
InnoPharma, Inc. |
New York |
NY |
US |
|
|
Assignee: |
InnoPharma, Inc.
New York
NY
|
Family ID: |
50233867 |
Appl. No.: |
15/478472 |
Filed: |
April 4, 2017 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
14023247 |
Sep 10, 2013 |
9636376 |
|
|
15478472 |
|
|
|
|
61699752 |
Sep 11, 2012 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/04 20180101;
A61K 9/0019 20130101; A61K 9/19 20130101; A61K 9/08 20130101; A61P
35/00 20180101; A61K 47/10 20130101; A61K 38/07 20130101; A61K
47/26 20130101 |
International
Class: |
A61K 38/07 20060101
A61K038/07; A61K 9/19 20060101 A61K009/19; A61K 47/10 20060101
A61K047/10; A61K 47/26 20060101 A61K047/26; A61K 9/08 20060101
A61K009/08; A61K 9/00 20060101 A61K009/00 |
Claims
1. A pharmaceutical composition, comprising: (i) a peptide epoxy
ketone or a pharmaceutically acceptable salt thereof; (ii) a
solvent system comprising a water miscible pharmaceutically
acceptable organic solvent suitable for injection selected from the
group consisting of ethanol, propylene glycol, polyethylene glycol,
and mixtures thereof, and optionally water; and (iii) a
non-volatile sugar acid.
2. The pharmaceutical composition of claim 1, wherein the
non-volatile sugar acid is selected from the group consisting of
lactobionic acid, glycolic acid, malic acid, citric acid, lactic
acid, mandelic acid, and tartaric acid.
3. The pharmaceutical composition of claim 2, wherein the peptide
epoxy ketone is carfilzomib or a pharmaceutically acceptable salt
thereof.
4. The pharmaceutical composition of claim 1, wherein the solvent
system comprises ethanol and propylene glycol.
5. The pharmaceutical composition of claim 1, wherein the solvent
system comprises ethanol and polyethylene glycol.
6. The pharmaceutical composition of claim 1, wherein peptide epoxy
ketone is carfilzomib or a pharmaceutically acceptable salt thereof
and the non-volatile sugar acid is lactobionic acid.
7. The pharmaceutical composition of claim 6, wherein the
concentration of the peptide epoxy ketone in the solvent system
ranges from about 0.1 to about 10.0 mg/mL and the weight ratio of
the peptide epoxy ketone to lactobionic acid ranges from about 2:1
to 15:1.
8. The pharmaceutical composition of claim 7, wherein the solvent
system comprises ethanol, propylene glycol, and water.
9. The pharmaceutical composition of claim 7, wherein the solvent
system comprises ethanol, polyethylene glycol, and water.
10. The pharmaceutical composition of claim 1 comprising: (i)
carfilzomib or the pharmaceutically acceptable salt thereof; (ii)
lactobionic acid, wherein the weight ratio of the lactobionic acid
to carfilzomib ranges from 2:1 to 15:1; and (iii) a solvent system
comprising ethanol, propylene glycol, and water, wherein the
ethanol and propylene glycol comprise more than about 50% (v/v) of
the solvent system, wherein the concentration of carfilzomib ranges
from about 1 to about 6 mg/mL.
11. The pharmaceutical composition of claim 1 comprising: (i)
carfilzomib or the pharmaceutically acceptable salt thereof; (ii)
lactobioic acid, wherein the weight ratio of the lactobionic acid
to carfilzomib ranges from about 4:1 to 7:1; and (iii) a solvent
system comprising ethanol, polyethylene glycol, and water, wherein
the ethanol and polyethylene glycol comprise more than about 50%
(v/v) of the solvent system, wherein the concentration of the
carfilzomib ranges from about 1 to about 6 mg/mL.
12. A pharmaceutical composition comprising a peptide epoxy ketone
and a sugar, wherein the pharmaceutical composition is
lyophilized.
13. The pharmaceutical composition of claim 12, wherein the sugar
acid is lactobionic acid.
14. The pharmaceutical composition of claim 12, wherein the peptide
epoxy ketone is carfilzomib.
15. The pharmaceutical composition of claim 14, sugar acid is
lactobionic acid.
16. The pharmaceutical composition of claim 13, wherein the weight
ratio of the peptide epoxy ketone to lactobionic acid ranges from
about 2:1 to 10:1.
17. The pharmaceutical composition of claim 15, wherein the weight
ratio of the lactobionic acid to carfilzomib ranges from about 2:1
to 10:1.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional
Application No. 61/699,752, filed Sep. 11, 2012, the specification
of which is hereby incorporated by reference in its entirety.
FIELD OF INVENTION
[0002] The present invention relates to pharmaceutical compositions
comprising a peptide epoxy ketone, which can be a proteasome
inhibitor, such as Carfilzomib, wherein the pharmaceutical
compositions provide improved stability and solubility for the
proteasome inhibitor contained therein.
BACKGROUND OF THE INVENTION
[0003] Carfilzomib, which is the active ingredient in
KYPROLIS.RTM., is a peptide epoxy ketone proteasome inhibitor
indicated for the treatment of patients with multiple myeloma who
have received at least two prior therapies, including bortezomib
and an immunomodulatory agent, and have demonstrated disease
progression on or within 60 days of completion of the last therapy.
Carfilzomib irreversibly binds to the N-terminal
threonine-containing active sites of the 20S proteasome, the
proteolytic core particle within the 26S proteasome. Carfilzomib
has been shown to have antiproliferative and proapoptotic
activities in vitro in solid and hematologic tumor cells. The
compound has also been shown to inhibit proteasome activity in
blood and tissue and delay tumor growth in models of multiple
myeloma, hematologic, and solid tumors.
[0004] Peptide epoxy ketone proteasome inhibitors, such as
Carfilzomib, have proven difficult to formulate due to their low
aqueous solubility. In addition, the proteasome inhibitors are very
unstable. Thus, compositions comprising these compounds must
typically be lyophilized before storage and reconstituted before
use. Moreover, the reconstituted compositions are often not stable
themselves (even under refrigerated conditions). As shown in Table
1, certain reconstituted KYPROLIS.RTM. compositions must be used
within 24 hours after reconstitution because of their
instability.
TABLE-US-00001 TABLE 1 Storage Conditions of Reconstituted
Stability*per Container KYPROLIS Vial Syringe IV Bag ( )
Refrigerated (2.degree. C. to 8.degree. C.; 24 hours 24 hours 24
hours 36.degree. F. to 46.degree. F.) Room Temperature (15.degree.
C. to 30.degree. C.; 4 hours 4 hours 4 hours 59.degree. F. to
86.degree. F.) *Total time from reconstitution to administration
should not exceed 24 hours. 5% Dextrose Injection, USP
[0005] The need to lyophilize a composition comprising a peptide
epoxy ketone proteasome inhibitor presents challenges to healthcare
professionals. For example, dosing errors frequently occur during
reconstitution of such products, and safety risks are presented if
the reconstitution is not conducted aseptically. Additionally, the
product after reconstitution is not stable for more than 24 hr, a
product with such stability and storage temperature restrictions
poses a problem for convenient use in a clinical use setting.
Furthermore, lyophilized products allow for a one time
reconstitution and use per patient. This type of presentation does
not allow for injecting multiple doses. Also any unused portion of
the reconstituted composition must be discarded, which leads to
drug wasting. For at least these reasons, there is a need for new
pharmaceutical compositions of peptide epoxy ketone proteasome
inhibitors that provide improved solubility and stability for the
peptide epoxy ketone proteasome inhibitor, as well as for
ready-to-use compositions that eliminate the need for
reconstitution.
SUMMARY OF THE INVENTION
[0006] Accordingly, in one aspect, the invention provides a
lyophilized composition comprising a peptide epoxy ketone and a
non-volatile sugar acid.
[0007] In one embodiment, the non-volatile sugar acid is
lactobionic acid.
[0008] In one embodiment, the peptide epoxy ketone is a proteasome
inhibitor.
[0009] In one embodiment, the peptide epoxy ketone is
Carfilzomib.
[0010] In one embodiment, the peptide epoxy ketone is Carfilzomib
and the non-volatile sugar acid is lactobionic acid.
[0011] The lyophilized pharmaceutical composition may be
administered to a patient in need of such compositions after the
composition is reconstituted with one or more pharmaceutically
acceptable diluents.
[0012] In another aspect, the invention provides a pharmaceutical
composition comprising (i) a peptide epoxy ketone; (ii) a solvent
system comprising one or more water miscible pharmaceutically
acceptable organic solvents suitable for injection, and optionally
water; and (iii) a non-volatile sugar acid.
[0013] In one embodiment, the water miscible pharmaceutically
acceptable organic solvent suitable for injection is selected from
the group consisting of ethanol, propylene glycol, polyethylene
glycol, glycerol, dimethylacetamide, N-methylpyrrolidone,
dimethylsulfoxide, and mixtures thereof.
[0014] In one embodiment, the water miscible pharmaceutically
acceptable organic solvent suitable for injection is selected from
the group consisting of ethanol, propylene glycol, polyethylene
glycol, and mixtures thereof
[0015] In one embodiment, the non-volatile sugar acid is
lactobionic acid.
[0016] In one embodiment, the peptide epoxy ketone is a proteasome
inhibitor.
[0017] In one embodiment, the peptide epoxy ketone is
Carfilzomib.
[0018] In one embodiment, the peptide epoxy ketone is Carfilzomib
and the non-volatile sugar acid is lactobionic acid.
[0019] In one embodiment, the peptide epoxy ketone is Carfilzomib;
the non-volatile sugar acid is lactobionic acid; and the water
miscible pharmaceutically acceptable organic solvent suitable for
injection is selected from the group consisting of ethanol,
propylene glycol, polyethylene glycol, and mixtures thereof
[0020] In one embodiment, the pharmaceutical composition is a ready
to use formulation.
[0021] In one embodiment, the pharmaceutical composition is a ready
to dilute formulation.
[0022] The pharmaceutical compositions of the invention may also
comprise one or more pharmaceutically acceptable excipients, such
as a buffer, surfactant, antioxidant, preservative, isotonicity
agent, and/or a lyoprotectant agent.
[0023] Another aspect of the invention relates to methods of making
the pharmaceutical compositions discussed herein.
[0024] The pharmaceutical compositions of the invention overcome
one or more limitations discussed above.
DETAILED DESCRIPTION OF THE INVENTION
[0025] The invention is generally directed towards pharmaceutical
compositions comprising a peptide epoxy ketone.
[0026] In one embodiment, the composition is a lyophilized
composition comprising the peptide epoxy ketone and a non-volatile
sugar acid.
[0027] In one embodiment, the composition is a liquid formulation
comprising (i) a peptide epoxy ketone; (ii) a solvent system
comprising one or more water miscible pharmaceutically acceptable
organic solvents suitable for injection, and optionally water; and
(iii) a non-volatile sugar acid.
[0028] In each of the embodiments, the peptide epoxy ketone can be
a proteasome inhibitor.
[0029] In each of the embodiments, the peptide epoxy ketone can be
Carfilzomib.
[0030] Where such compositions are lyophilized or concentrated
above the concentration suitable for injection or infusion, the
compositions will be administered after reconstitution with one or
more pharmaceutically acceptable diluents.
[0031] Regardless of the particular form of the preparation, the
pharmaceutical compositions contemplated herein may further
comprise one or more buffers, surfactants, antioxidants,
preservatives, isotonicity agents, and/or lyoprotectants.
[0032] Pharmaceutical compositions according to the instant
disclosure provide improved stability and solubility for peptide
epoxy ketones, such as Carfilzomib.
[0033] Any recitation of ranges of values set forth below is merely
intended to serve as a shorthand method of referring individually
to each separate value falling within the range, unless otherwise
indicated herein, and each separate value is incorporated into the
specification as if it were individually recited herein.
Furthermore, all references, including publications, patent
applications, and patents, cited herein are hereby incorporated by
reference to the same extent as if each reference were individually
and specifically indicated to be incorporated by reference and were
set forth in its entirety herein.
[0034] The terms "a" and "an" and "the," as used herein, are to be
construed to cover both the singular and the plural, unless
otherwise indicated herein or clearly contradicted by context.
[0035] The terms "comprising," "having," "including," and
"containing," as used herein, are to be construed as open-ended
terms (i.e., meaning "including, but not limited to,") unless
otherwise noted.
[0036] The pharmaceutical compositions of the invention comprise at
least one peptide epoxy ketone or a pharmaceutically acceptable
salt thereof. Preferably, the peptide epoxy ketone is a proteasome
inhibitor. Peptide epoxy ketones are epoxide-containing compounds,
which contain a ketone group proximate to an epoxide group. Peptide
epoxy ketones have the general structure:
##STR00001##
wherein:
[0037] R.sup.1 and R.sup.2 are hydrogen or a C.sub.1-C.sub.8 alkyl
group, such as methyl, ethyl, propyl and butyl groups, which can be
further substituted with one or more hydroxy, halogen, amino,
carboxy, carbonyl, thio, sulfide, ester, amide, or C.sub.1-C.sub.6
alkoxy groups; and
[0038] P is a peptide chain containing between 1 and 12 amino acids
that is connected to the nitrogen by an amide bond with the
carboxylic acid terminus of the amino acid. The N-terminal of the
peptide chain nay be protected or derivatized.
[0039] The stereochemistry of the carbons bonded to R.sup.1 and
R.sup.2 can be (R) or (S). In one preferred embodiment the
stereochemistry of the carbon bonded to R.sup.1 and R.sup.2 is as
depicted below:
##STR00002##
For some embodiments, the oxygen atom of the epoxide is configured
as depicted above in order to facilitate interaction with an
N-terminal nucleophilic group in an Ntn hydrolase. For example,
irreversible interactions of enzyme inhibitors with the
.beta.5/Pre2 subunit of 20S proteasome, which lead to inhibition,
appear to be facilitated by the configuration illustrated above. In
the case of other Ntn hydrolases, the opposite stereochemistry of
the epoxide may be preferred.
[0040] Preferably, the peptide chain contains two or more amino
acids. In some embodiments, useful for inhibiting chymotrypsin-like
(CT-L) activity of the proteasome, the peptide chain contains
between two and eight amino acids, and in some preferred
embodiments for CT-L inhibition, between two and six amino acids.
In other embodiments useful for inhibiting the PGPH activity of the
proteasome, the peptide chain contains between two and eight amino
acids, and in some preferred embodiments for PGPH inhibition,
between two and six amino acids. In other embodiments, the peptide
chain contains between two and four amino acid units.
[0041] The amino acids can be a natural occurring essential amino
acid, such as glycine, alanine, valine, isoleucine, leucine,
phenylalanine, threonine, tyrosine, serine, and proline. The amino
acids can be a naturally occurring non-essential amino acid, such
as for example taurine, carnitine, citrulline, cystine, omithine,
norleucine, and others. The amino acids can also be a non-naturally
occurring amino acid of general formula:
##STR00003##
wherein the side chain, R, can be hydrogen, phenyl, or
C.sub.1-C.sub.8 alkyl, wherein the C.sub.1-C.sub.8 alkyl can be
further substituted with one or more phenyl, hydroxy, halogen,
amino, carboxy, carbonyl, thio, sulfide, ester, amide, or
C.sub.1-C.sub.6 alkoxy groups, and the phenyl can be further
substituted with one or more hydroxy, halogen, amino, carboxy,
carbonyl, thio, sulfide, ester, amide, or C.sub.1-C.sub.6 alkoxy
groups.
[0042] Peptide epoxy ketones useful in the compositions of the
invention include those described in U.S. Pat. No. 7,737,112, the
content of which are incorporated herein by reference.
[0043] Most preferably, the peptide epoxy ketone proteasome
inhibitor is Carfilzomib (i.e.,
(2S)-N-((S)-1-(S)-4-methyl-1-((R)-2-methyloxiran-2-yl)-1-oxopentan-2-ylca-
rbamoyl)-2-phenylethyl)-2-((S)-2-(2-morpholinoacetamido)-4-phenylbutanamid-
o)-4-methylpentanamide) which has the following structure:
##STR00004##
[0044] Lyophilized Compositions:
[0045] The lyophilized compositions of the invention comprise the
peptide epoxy ketone and a non-volatile sugar acid. The peptide
epoxy ketone may be a proteasome inhibitor. Preferably, the peptide
epoxy ketone is Carfilzomib.
[0046] The weight ratio of the non-volatile sugar acid to the
peptide epoxy ketone ranges from about 2:1 to 15:1, preferably
about 3:1 to 15:1, more preferably about 4:1 to 15:1, most
preferably about 4:1 to 10:1. Typically, the weight ratio of the
sugar acid to the peptide epoxy ketone is greater than 4:1,
preferably greater than 5:1. Typically, the weight ratio of the
sugar acid to the peptide epoxy ketone is less than 15:1,
preferably less than 10:1.
[0047] In one embodiment, the lyophilized composition comprises
about 60 mg of the peptide epoxy ketone and about 300 mg of the
non-volatile sugar acid. In one embodiment, the lyophilized
composition comprises about 60 mg of Carfilzomib and about 300 mg
of lactobionic acid. In one embodiment, the lyophilized composition
comprises about 60 mg of Carfilzomib and about 400 mg of
lactobionic acid.
[0048] The phrase sugar acid means an oxidized derivative of sugar
having one or more carboxylic acid functional groups. Sugar acids
can be obtained by oxidizing the aldehyde group of a sugar. The
term sugar includes any polyhydroxy carbohydrate moiety, including
monosaccharides, disaccharides, polysaccharides. Preferably, the
sugar acid is derived from a monosaccharide or a disaccharide. In
one embodiment the sugar acid is a monosaccharide. In one
embodiment the sugar acid is a disaccharide. The sugar acids may be
used in the acid form or as a salt. When used as a salt, the salt
is preferably a sodium, calcium, magnesium, or zinc salt. The sugar
acid may be in the form of an ester or a lactone. Sugar acids also
include compounds derived from amino sugars and acetylated amino
sugars.
[0049] Non-volatile sugar acids useful in the compositions of the
invention include, but are not limited to, N-acetylneuraminic acid,
N-acetyltalosaminuronic acid, aldaric acid, aldonic acid,
3-deoxy-D-manno-oct-2-ulosonic acid, galaturonic acid,
D-galacturonic acid, glucaric acid, gluconic acid, glucuronic acid,
glucono-.gamma.-lactone, glyceric acid, N-glycolylneuraminic acid,
iduronic acid, isosaccharinic acid, lactobionic acid, mucic acid,
muramic acid, neuraminic acid, pangamic acid, saccharic acid,
sialic acid, threonic acid, ulosonic acid, uronic acid, X-Gluc,
xylonic acid, ascorbic acid, and mixtures thereof. Preferably, the
non-volatile organic acid is lactobionic acid.
[0050] Preferred sugar acids include, but are not limited to,
gluconic acid, glucono-.gamma.-lactone, lactobionic acid,
glucuronic acid and its mono-or dilactones, pangamic acid,
mannosaccharic acid and its mono-or dilactones, mucic acid and its
mono-or dilactones, and mixtures thereof. Most preferred sugar acid
is lactobionic acid.
[0051] The lyophilized compositions may further include one or more
excipients suitable for inclusion in a lyophilized composition. For
example, the lyophilized compositions may include a lyoprotectant.
Suitable lyoprotectants include amino acids and polymers.
Preferably, the amino acid is selected from the group consisting of
lysine, alanine, and glycine. Suitable polymers include various
proteins (e.g., gelatin and albumin), polyethylene glycol, gelatin,
polyvinyl pyrrolidone, albumin, and Dextran-40. Typically the
lyoprotectant represents less than 50% weight/weight of the
lyophilized composition, and all concentrations above 1%
weight/weight of the total composition are deemed effective to
enhance the stability of the composition. In various embodiments,
the lyoprotectant is present in an amount of at least about 5%
weight/weight, at least about 10% weight/weight, or at least about
20% weight/weight of the total composition.
[0052] The compositions can be prepared by combining the peptide
epoxy ketone, the non-volatile sugar acid, any optional excipients,
and a suitable lyophilization solvent to provide a solution and
then lyophilizing the solution using art-recognized lyophilization
techniques.
[0053] The compositions can be sterilized. Sterilized compositions
can be obtained using art-recognized methods for sterilization,
such as filtration through 0.22 micron filters (e.g., PVDF
filters), heat sterilization, radiation (e.g., gamma, electron
beam, microwave), and/or ethylene oxide sterilization.
[0054] The non-volatile sugar acid compositions are more stable
than a comparable composition that does not include the
non-volatile sugar acid. Preferably the compositions have
sufficient stability to allow storage at a commercially relevant
temperature, such as between about 0.degree. C. and about
60.degree. C., for a commercially relevant period of time, such as
at least one week, preferably at least one month, more preferably
at least three months, and most preferably at least six months. In
some embodiments the compositions have sufficient stability to
allow storage at a commercially relevant temperature for more than
one year, and in some embodiments more than two years. Stability
can be measured using any physiochemical characterization
techniques known to those skilled in the art, such as, for example
high pressure liquid chromatography (HPLC).
[0055] The lyophilized pharmaceutical compositions discussed herein
may be in a lyophilized powder or lyophilized cake form. Before
use, the lyophilized pharmaceutical compositions are diluted or
reconstituted with a pharmaceutically acceptable diluent. Any
diluent known in the art in which the peptide epoxy ketone is
soluble can be used. Preferably, the lyophilized composition is
reconstituted with a solvent system so as to provide a liquid
pharmaceutical composition as described below. In one embodiment,
reconstitution provides a ready to use formulation as described
below. In one embodiment, reconstitution provides a ready to dilute
formulation as described below. The ready to dilute formulation can
then be diluted with a suitable diluent before administration.
Suitable diluents include, but are not limited to, water, saline,
dextrose 5% in water, water for injection, and lactated ringer's
solution. In one embodiment, the lyophilized composition is
reconstituted directly with a suitable diluent.
[0056] The lyophilized pharmaceutical compositions are preferably
contained in a vial or pre-filled syringe before reconstitution
occurs.
[0057] It should be appreciated that the peptide epoxy ketone is
present in the lyophilized pharmaceutical compositions in an amount
that is suitable for administration after reconstitution.
[0058] Liquid Pharmaceutical Compositions:
[0059] The liquid pharmaceutical compositions of the invention
comprise (i) a peptide epoxy ketone, (ii) a solvent system
comprising one or more water miscible pharmaceutically acceptable
organic solvents suitable for injection and optionally water, and
(iii) a non-volatile sugar acid.
[0060] In one embodiment, the liquid pharmaceutical composition is
a ready to use formulation. A ready to use formulation is a
formulation that is suitable for administration without further
dilution.
[0061] In one embodiment, the liquid pharmaceutical composition is
a ready to dilute formulation. A ready to dilute formulation is a
formulation that is suitable for administration after dilution with
a suitable diluent.
[0062] The liquid pharmaceutical compositions can be a multi-dose
formulation, i.e., a formulation that has a volume and/or quantity
of the active pharmaceutical ingredient suitable for at least two
independent and distinct administrations (to the same or a
different patient) of the formulation.
[0063] The liquid pharmaceutical compositions are more stable than
a comparable composition that does not include the non-volatile
sugar acid. Without wishing to be bound by theory, it is believed
that the sugar acid provides improved stability to the liquid
pharmaceutical compositions. Importantly, the sugar acid also helps
prevent the peptide epoxy ketone from precipitating when the liquid
pharmaceutical composition is further diluted with a diluent. Thus,
the liquid pharmaceutical compositions can advantageously be more
readily diluted with a suitable diluent, without the peptide epoxy
ketone precipitating. By reducing the tendency of the peptide epoxy
ketone to precipitate, permits the preparation of more concentrated
formulations to be obtained by diluting the by liquid
pharmaceutical compositions.
[0064] The water miscible pharmaceutically acceptable organic
solvent suitable for injection provides improved solubility for the
peptide epoxy ketone compared to an aqueous composition in the
absence of the pharmaceutically acceptable organic solvent. The
pharmaceutically acceptable organic solvent also provides a solvent
system that reduces degradation of the peptide epoxy ketone
compared to an aqueous solvent in the absence of the organic
solvent
[0065] In one embodiment, the peptide epoxy ketone is a proteasome
inhibitor. In one embodiment, the peptide epoxy ketone is
Carfilzomib.
[0066] Typically, the concentration of the peptide epoxy ketone in
the liquid pharmaceutical composition ranges from about 0.1 to
about 10.0 mg/mL. For example, the concentration of the peptide
epoxy ketone in a ready to use formulation ranges typically from
about 0.1 to about 3.0 mg/mL and the concentration of the peptide
epoxy ketone in a ready to dilute formulation typically ranges from
about 1.0 to about 10.0 mg/mL.
[0067] Any of the non-volatile sugar acids described above can be
used in the liquid pharmaceutical compositions. The weight ratio of
the non-volatile sugar acid to the peptide epoxy ketone ranges from
about 2:1 to 15:1, preferably about 3:1 to 15:1, more preferably
about 4:1 to 15:1, most preferably about 4:1 to 10:1. Typically,
the weight ratio of the non-volatile sugar acid to the peptide
epoxy ketone is greater than 4:1, preferably greater than 5:1.
Typically, the weight ratio of the non-volatile sugar acid to the
peptide epoxy ketone is less than 15:1, preferably less than
10:1.
[0068] Preferably, the non-volatile sugar acid is lactobionic
acid.
[0069] Suitable water miscible pharmaceutically acceptable organic
solvents suitable for injection are non-toxic organic solvents in
which the peptide epoxy ketone is soluble. Suitable solvents
include of ethanol, propylene glycol, polyethylene glycol,
glycerol, dimethylacetamide, N-methylpyrrolidone,
dimethylsulfoxide, and mixtures thereof In one embodiment, the
solvent is selected from the group consisting of ethanol, propylene
glycol, polyethylene glycol, and mixtures thereof.
[0070] Preferably, the water miscible pharmaceutically acceptable
organic solvent suitable for injection is selected from the group
consisting of ethanol, propylene glycol, polyethylene glycol (in
particular PEG 300 and PEG 400), and mixtures thereof
[0071] The liquid pharmaceutical compositions may include water.
Preferably the water miscible pharmaceutically acceptable organic
solvent suitable for injection comprises more than about 25% by
volume of the total amount of liquid used to formulate the
pharmaceutical composition. More preferably, the water miscible
pharmaceutically acceptable organic solvent suitable for injection
comprises more than about 50% by volume of the total amount of
liquid used to formulate the pharmaceutical composition. Most
preferably, the water miscible pharmaceutically acceptable organic
solvent suitable for injection comprises more than about 75% by
volume of the total amount of liquid used to formulate the
pharmaceutical compositions
[0072] In one embodiment, the liquid pharmaceutical composition
comprises at least two pharmaceutically acceptable solvents. The at
least two pharmaceutically acceptable solvents can be present in
any ratio.
[0073] In one embodiment, the liquid pharmaceutical composition
comprises both ethanol and propylene glycol. Preferably, the volume
ratio of ethanol to propylene glycol ranges from about 1:10 to
10:1. In one embodiment, the liquid pharmaceutical composition
comprises ethanol, propylene glycol, and water. In one embodiment,
the volume percent of the water ranges from 0% to about 60% of the
total volume of liquid in the composition.
[0074] In one embodiment, the liquid pharmaceutical composition
comprises both ethanol and polyethylene glycol (e.g., PEG 300 or
PEG 400). Preferably, the volume ratio of ethanol to polyethylene
glycol ranges from about 1:10 to 10:1. In one embodiment, the
liquid pharmaceutical composition comprises ethanol, polyethylene
glycol, and water. In one embodiment, the volume percent of the
water ranges from 0% to about 60% of the total volume of liquid in
the composition.
[0075] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib; a solvent system comprising ethanol,
propylene glycol, and water; and a non-volatile sugar acid.
[0076] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib; a solvent system comprising ethanol,
propylene glycol, and water; and lactobionic acid.
[0077] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib in an amount of about 0.1 to 10 mg/mL; a
solvent system comprising ethanol, propylene glycol, and water; and
lactobionic acid. In one embodiment, the Carfilzomib is present in
an amount of about 1 to 5 mg/mL.
[0078] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib in an amount of about 0.1 to 10 mg/mL; a
solvent system comprising ethanol, propylene glycol, and water; and
lactobionic acid, wherein the weight ratio of lactobionic acid to
Carfilzomib ranges from about 2:1 to about 15:1. In one embodiment,
the Carfilzomib is present in an amount of about 1 to 5 mg/mL.
[0079] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib in an amount of about 0.1 to 10 mg/mL; a
solvent system comprising ethanol, propylene glycol, and water; and
lactobionic acid, wherein the weight ratio of lactobionic acid to
Carfilzomib ranges from about 2:1 to about 15:1, and the ratio of
propylene glycol to ethanol is about 1:10 to 10:1 (v/v). In one
embodiment, the ethanol and propylene glycol comprise more than
about 25% by volume of the total amount of liquid used to formulate
the pharmaceutical composition. In one embodiment, the ethanol and
propylene glycol comprise more than about 50% by volume of the
total amount of liquid used to formulate the pharmaceutical
composition. In one embodiment, the ethanol and propylene glycol
comprise more than about 75% by volume of the total amount of
liquid used to formulate the pharmaceutical composition. In one
embodiment, the Carfilzomib is present in an amount of about 1 to 5
mg/mL.
[0080] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib; a solvent system comprising ethanol,
polyethylene glycol, and water; and a non-volatile sugar acid. In
one embodiment, the polyethylene glycol is selected from PEG 300
and PEG 400.
[0081] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib; a solvent system comprising ethanol,
polyethylene glycol, and water; and lactobionic acid. In one
embodiment, the polyethylene glycol is selected from PEG 300 and
PEG 400.
[0082] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib in an amount of about 0.1 to 10 mg/mL; a
solvent system comprising ethanol, polyethylene glycol, and water;
and lactobionic acid. In one embodiment, the polyethylene glycol is
selected from PEG 300 and PEG 400. In one embodiment, the
Carfilzomib is present in an amount of about 1 to 5 mg/mL.
[0083] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib in an amount of about 0.1 to 10 mg/mL; a
solvent system comprising ethanol, polyethylene glycol, and water;
and lactobionic acid, wherein the weight ratio of lactobionic acid
to Carfilzomib ranges from about 2:1 to about 15:1. In one
embodiment, the polyethylene glycol is selected from PEG 300 and
PEG 400.
[0084] In one embodiment, the liquid pharmaceutical composition
comprises Carfilzomib in an amount of about 0.1 to 10 mg/mL; a
solvent system comprising ethanol, polyethylene glycol, and water;
and lactobionic acid, wherein the weight ratio of Carfilzomib to
lactobionic acid ranges from about 2:1 to about 15:1, and the ratio
of ethanol to polyethylene glycol is about 1:10 to 10:1 (v/v). In
one embodiment, the ethanol and polyethylene glycol comprise more
than about 25% by volume of the total amount of liquid used to
formulate the pharmaceutical composition. In one embodiment, the
ethanol and polyethylene glycol comprise more than about 50% by
volume of the total amount of liquid used to formulate the
pharmaceutical composition. In one embodiment, the ethanol and
polyethylene glycol comprise more than about 75% by volume of the
total amount of liquid used to formulate the pharmaceutical
composition. In one embodiment, the Carfilzomib is present in an
amount of about 1 to 5 mg/mL.
[0085] The liquid pharmaceutical compositions of the instant
invention may further comprise one or more excipients, such as,
buffers, surfactants, antioxidants, preservatives, isotonicity
agents, and/or lyoprotectants.
[0086] For instance, a buffer is typically used in the liquid
pharmaceutical compositions of the invention to control the pH. As
shown in Table 2, at least some peptide epoxy ketones (e.g.,
Carfilzomib) have demonstrated a strong pH-dependent
solubility.
TABLE-US-00002 TABLE 2 Effect of pH on Solubility of Carfilzomib pH
Saturation Solubility (.mu.g/mL) 1.7 (1% L-Cysteine) 459.0 3.5 (15
mM Acetate Buffer) 125.0 5.0 (15 mM Citrate Buffer) 9.0 7.0 (15 mM
Phosphate Buffer) 5.0
Accordingly, a buffer may be used in the liquid pharmaceutical
compositions of the instant invention to maintain a pre-selected pH
level at which the peptide epoxy ketone is soluble.
[0087] Examples of suitable buffers include mixtures of a weak acid
and alkali metal salt (e.g., sodium, potassium) and the conjugate
base of the weak acid. Suitable buffers include, for example,
buffers selected from the group consisting of citric acid, acetic
acid, maleic acid, phosphoric acid, succinic acid, or tartaric
acid, as well as the counter ion salts thereof. The molar
concentration of the buffer typically ranges between about 5
millimolar and about 150 millimolar, and is most preferably about
15 millimolar. The buffer will typically have a pH of about 3.5,
however, modifications to the pH value are also contemplated
herein. It will be recognized by those of skill in the art that
determination of the preferred solubility range of a peptide epoxy
ketone around a pH of 3.5 using any of the aforementioned buffers
can be performed using known techniques.
[0088] The pH level for each pharmaceutical composition should be
selected to provide suitable solubility of the peptide epoxy ketone
used therein. It is generally preferred, however, that the pH of
the compositions be suitable for injection and, therefore, will
typically be between about 2.0 and about 9.0, and even more
typically between about 2.5 and about 8.0.
[0089] The liquid pharmaceutical compositions according the
invention may also comprise an anionic, nonionic, cationic, or
amphoteric surfactant. Anionic surfactants that may be used in the
pharmaceutical compositions of the invention include alkyl ether
sulfates and carboxylic acids, in particular in form of their
alkali salts, as well as protein fatty acid condensates.
[0090] Examples of suitable nonionic surfactants include, but are
not limited to, fatty alcohol acid or amide ethoxylates,
monoglyceride ethoxylates, sorbitan ester ethoxylates alkyl
polyglycosides, mixtures thereof, and the like. Certain preferred
nonionic surfactants include polyoxyethylene derivatives of polyol
esters, wherein the polyoxyethylene derivative of polyol ester (1)
is derived from (a) a fatty acid containing from about 8 to about
22, and preferably from about 10 to about 14 carbon atoms, and (b)
a polyol selected from sorbitol, sorbitan, glucose, .alpha.-methyl
glucoside, polyglucose having an average of about 1 to about 3
glucose residues per molecule, glycerine, pentaerythritol and
mixtures thereof, (2) contains an average of from about 10 to about
120, and preferably about 20 to about 80 oxyethylene units; and (3)
has an average of about 1 to about 3 fatty acid residues per mole
of polyoxyethylene derivative of polyol ester. Examples of such
preferred polyoxyethylene derivatives of polyol esters include, but
are not limited to PEG-80, sorbitan laurate, and Polysorbate
80.
[0091] Another class of suitable nonionic surfactants includes long
chain alkyl glucosides or polyglucosides, which are the
condensation products of (a) a long chain alcohol containing from
about 6 to about 22, and preferably from about 8 to about 14 carbon
atoms, with (b) glucose or a glucose-containing polymer. Preferred
alkyl gluocosides comprise from about 1 to about 6 glucose residues
per molecule of alkyl glucoside. A preferred glucoside is decyl
glucoside, which is the condensation product of decyl alcohol with
a glucose polymer and is available commercially from Cognis
Corporation of Ambler, Pa. under the tradename, "Plantaren
2000."
[0092] Various cationic surfactants may also be suitable for use in
the present compositions. Examples of suitable cationic surfactants
include, but are not limited to alkyl quaternaries (mono, di, or
tri), benzyl quaternaries, ester quaternaries, ethoxylated
quaternaries, alkyl amines, and mixtures thereof, wherein the alkyl
group has from about 6 carbon atoms to about 30 carbon atoms, with
about 8 to about 22 carbon atoms being preferred.
[0093] Also possible is the incorporation of amphoteric
surfactants, such as alkyl betaines, alkyl amido betaines, and
alkyl amphoacetates.
[0094] To still further improve the stability, the liquid
pharmaceutical compositions may also include one or more
anti-oxidants. For example, hydrophobic anti-oxidants include
butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate,
and .alpha..-tocopherol, DL-tocopherol, .alpha.-tocopherol acetate,
.alpha.-tocopherol Tocopherol Polyethylene Glycol Succinate
(Vitamin E TPGS), L-cysteine, or hydrophilic anti-oxidants,
including sodium EDTA and thioglycerol. Most typically, the
concentration of the anti-oxidant will be between 0.005% and 5%
weight/weight of the total composition.
[0095] Additionally, or alternatively, the contemplated liquid
pharmaceutical compositions may include a preservative (e.g.,
phenol, thimerosal, chlorobutanol, benzyl alcohol, m-cresol,
phenoxyethanol, methylparaben and propylparaben), typically at a
concentration of between 0.001% weight/weight and about 5%
weight/weight of the total composition, and is most typically
between about 0.003% and about 2.0% weight/weight of the total
composition.
[0096] The liquid pharmaceutical compositions contemplated herein
may further include isotonicity agents and/or lyoprotectants.
Suitable isotonicity agents include sodium chloride, glycerol, and
thioglycerol. Suitable lyoprotectants are described above.
[0097] The liquid pharmaceutical compositions can be prepared
according to a method comprising the steps:
[0098] (a) placing a desired amount of a peptide epoxy ketone in a
compounding vessel;
[0099] (b) adding one or more water miscible pharmaceutically
acceptable organic solvents suitable for injection to the
compounding vessel;
[0100] (c) optionally adding water to the compounding vessel;
[0101] (d) mixing the contents of the compounding vessel;
[0102] (e) optionally adding one or more excipients to the
compounding vessel;
[0103] (f) adding a non-volatile sugar acid to the compounding
vessel; and
[0104] (g) mixing the contents of the compounding vessel until the
non-volatile sugar acid is dissolved.
[0105] The contents of the compounding vessel may be sterilized
using any known methods of sterilization, including filtration
through 0.22 micron filters (e.g., PVDF filters), heat
sterilization, radiation (e.g., gamma, electron beam, microwave),
and/or ethylene oxide sterilization.
[0106] The pharmaceutical compositions of the instant invention are
suitable for administration to mammals, including humans. When
administered to humans, the compositions are typically administered
by intramuscular, subcutaneous, intra-arterial, or oral routes.
[0107] Regardless of the particular composition, it is preferred
that the composition is packaged in a container suitable for single
or multi-use. Such containers include an ampoule, a vial, a
pre-filled syringe, and an intravenous bag. Multi-use containers
may contain the peptide epoxy ketone in an amount suitable to allow
at least two distinct uses, more typically at least five distinct
uses, and even more typically at least ten distinct uses (each use
which may or may not require the same quantity of composition
administered to the patient). Thus, preferred multi-use containers
will be configured to contain a volume of the composition that is
suitable for multiple and independent administrations. Such
containers include vials with a rubber stopper that can be pierced
with a needle of a syringe.
[0108] The following examples are presented for illustrative
purposes only, and are not intended to limit the scope of the
invention.
EXAMPLES
[0109] For each of the Examples described below, the following HPLC
methodology was used to determine the relative retention time (RRT)
of impurities in the compositions:
[0110] Mobile Phase A: 0.5 mL of TFA in 1000 mL of distilled
water
[0111] Mobile Phase B: 100% HPLC grade Acetonitrile
[0112] Diluent: Acetonitrile: Water (1:1 v/v)
[0113] Elution program:
TABLE-US-00003 Time Min Flow mL % A % B 0.01 1.00 95.0 5.0 5.00
1.00 90.0 10.0 10.00 1.00 85.0 15.0 38.00 1.00 20.0 80.0 40.00 1.00
0.0 100.0 43.00 1.00 0.0 100.0 45.00 1.00 95.0 5.0 50.00 1.00 95.0
5.0
[0114] Flow rate: 1.0 mL/min
[0115] Detector Wavelength: 210 nm
[0116] Injection volume: 10 micro liters
[0117] Run time: 50 min
[0118] Column: Waters Symmetry C18 (250.times.4.6 mm), 5
microns
[0119] Sample temperature: 5.degree. C.
[0120] Column temperature: 25.degree. C.
Example 1
Ready-to-Dilute Composition (60 mg/Vial) to be Diluted Before
Injecting
TABLE-US-00004 [0121] Ingredient Amount Carfilzomib 60 milligrams
Ethanol 5 milliliters Propylene Glycol 2.5 milliliters Lactobionic
Acid 400 milligrams Acetate Buffer (15 millimolar, pH 3.5) 1
milliliter Polysorbate 80 500 milligrams 5% Dextrose 0.5
milliliters
[0122] The composition of Example 1 was prepared by placing 60
milligrams of Carfilzomib in a compounding vessel. 5 milliliters of
ethanol was then added to the compounding vessel and the contents
of the compounding vessel were mixed. 2.5 milliliters propylene
glycol was added to the compounding vessel and the contents of the
vessel were again mixed. Next, 1 milliliter of 15 mM acetate buffer
and 500 milligrams of Polysorbate 80 were added to the compounding
vessel and the contents therein were mixed. 400 milligrams
lactobionic acid was then added to the compounding vessel and the
contents of the vessel were mixed until the lactobionic acid was
dissolved. 0.5 milliliters of a 5% dextrose solution was then added
to the compounding vessel and the contents therein were mixed. Once
mixing was completed, the contents of the compounding vessel were
filtered through a 0.22 micron PVDF filter.
[0123] Physicochemical data for the composition of Example 1 is
shown below:
TABLE-US-00005 Impurities Assay RRT RRT RRT RRT Temperature Time
Appearance (%) 0.82 0.98 1.02 1.18 Total T = 0 CCS 104.9 -- 0.79
0.77 -- 1.56 40.degree. C./75% RH 1 Week CCS 89.3 0.05 1.26 0.77 --
2.05 2 Week CCS 113.4 0.57 2.11 0.85 -- 3.53 1 Month CCS 90.6 0.10
2.67 0.64 4.23 7.64 3 Month CCS 44.64 0.28 4.07 0.28 16.15 20.78
25.degree. C./60%RH 1 Month CCS 105.8 0.15 1.11 0.77 -- 2.03 3
Month CCS 107.9 0.63 2.13 0.73 1.22 4.71 CCS = clear colorless
solution RRT = relative retention time
[0124] As can be seen from the physicochemical data, the
composition of Example 1, which comprises lactobionic acid as well
as ethanol and propylene glycol solvents, remained stable for an
extended period of time even when stored at temperatures of
25.degree. Celsius. The ready to dilute composition is stable at
room temperature for at least 3 months.
Example 2
Ready-to-Dilute Composition (60 mg/Vial) to be Diluted Before
Injecting
TABLE-US-00006 [0125] Ingredient Amount Carfilzomib 60 milligrams
Ethanol 5 milliliters PEG 300 2.5 milliliters Lactobionic Acid 400
milligrams Acetate Buffer (15 millimolar, pH 3.5) 1 milliliter
Polysorbate 80 500 milligrams 5% Dextrose 0.5 milliliters
[0126] The composition of Example 2 was prepared by placing 60
milligrams of Carfilzomib in a compounding vessel. 5 milliliters of
ethanol was then added to the compounding vessel and the contents
of the compounding vessel were mixed. 2.5 milliliters PEG 300 was
added to the compounding vessel and the contents of the vessel were
again mixed. Next, 1 milliliter of 15 mM acetate buffer, and 500
milligrams of Polysorbate 80, were added to the compounding vessel
and the contents therein were mixed. 400 milligrams lactobionic
acid was then added to the compounding vessel and the contents of
the vessel were mixed until the lactiobionic acid was dissolved.
0.5 milliliters of a 5% dextrose solution was then added to the
compounding vessel and the contents therein were mixed. Once mixing
was completed, the contents of the compounding vessel were filtered
through a 0.22 micron PVDF filter.
[0127] Physicochemical data of the composition of Example 2 is
shown below:
TABLE-US-00007 Impurities Assay RRT RRT RRT RRT Temperature Time
Appearance (%) 0.82 0.98 1.02 1.18 Total T = 0 CCS 97.0 -- 0.78
0.77 -- 1.55 40.degree. C./75% RH 1 Week CCS 98.1 0.34 1.35 0.77 --
2.45 2 Week CCS 87.0 0.66 1.96 0.71 -- 3.33 1 Month CCS 84.8 1.33
2.83 0.57 2.93 7.66 3 Month CCS 53.0 0.22 4.19 0.37 8.74 13.52
25.degree. C./60% RH 1 Month CCS 106.0 0.22 1.14 0.75 -- 2.08 3
Month CCS 99.65 0.84 2.14 0.65 0.66 4.29 CCS = clear colorless
solution PPT = precipitate RRT = relative retention time
[0128] As can be seen from the physicochemical data, the
composition of Example 2, which comprises lactobionic acid as well
the solvents ethanol and PEG 300, remained stable for an extended
period of time even when stored at temperatures of 25.degree.
Celsius. The ready to dilute composition is stable at room
temperature for at least 3 months.
Example 3
Ready-to-Use Composition (2 mg/mL)
TABLE-US-00008 [0129] Ingredient Amount Carfilzomib 60 mg Ethanol
12 mL PEG 300 2.5 mL Lactobionic Acid 400 mg Acetate Buffer (15
millimolar, pH 3.5) 2 mL Polysorbate 80 500 mg 5% Dextrose Q.S to
30 g
[0130] The composition of Example 3 was prepared by placing 60
milligrams of Carfilzomib in a compounding vessel. 12 milliliters
of ethanol was then added to the compounding vessel and the
contents of the compounding vessel were mixed. 2.5 milliliters PEG
3000 was added to the compounding vessel and the contents of the
vessel were again mixed. Next, 2 mL of 15 mM acetate buffer and 500
milligrams of Polysorbate 80 were added to the compounding vessel
and the contents therein were mixed. 400 milligrams lactobionic
acid was then added to the compounding vessel and the contents of
the vessel were mixed until the lactobionic acid was dissolved. A
sufficient amount of 5% dextrose solution was then added to the
compounding vessel so as to provide 30 g and the contents therein
were mixed. Once mixing was completed, the contents of the
compounding vessel were filtered through a 0.22 micron PVDF
filter.
[0131] Physicochemical data for the composition of Example 3 is
shown below:
TABLE-US-00009 Impurities Assay RRT RRT RRT RRT Temperature Time
Appearance (%) 0.82 0.98 1.02 1.18 Total T = 0 CCS 107.5 -- 0.85
0.77 -- 1.61 40.degree. C./75% RH 1 Week CCS 100.9 0.47 1.7 0.77 --
2.91 2 Week CCS 96.2 0.82 2.47 0.75 4.03 1 Month CCS 88.4 1.79 3.75
0.6 -- 6.15 2 Month CCS 105.5 0.39 1.21 0.64 -- 2.23 25.degree.
C./60% RH 1 Month CCS 108.1 0.48 1.32 0.75 -- 2.55 2 Month CCS
99.31 0.96 2.53 0.56 -- 4.04 3 Month CCS 97.46 1.07 2.70 0.55 --
4.31 4.degree. C. 2 Month CCS 113.17 0.41 1.33 0.69 -- 2.43 3 Month
CCS 108.29 -- 0.79 0.66 -- 1.44 CCS = clear colorless solution RRT
= relative retention time
[0132] The ready to use composition is stable at room temperature
for at least 3 months.
[0133] The preceding examples, and exemplary language, are merely
illustrate and should not be construed to limit the scope of the
invention discussed herein, unless otherwise claimed. In addition,
no language in the specification should be construed as indicating
any non-claimed element as essential to the practice of the
invention. For instance, while particular embodiments of the
present invention have been illustrated and described, it should be
apparent to those skilled in the art that various other changes and
modifications can be made without departing from the spirit and
scope of the invention. For instance, all methods described herein
can be performed in any suitable order unless otherwise indicated
herein or otherwise clearly contradicted by context.
* * * * *