Note: Descriptions are shown in the official language in which they were submitted.
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PHARMACEUTICAL COMPOSITIONS FOR VEIN IRRITATING DRUGS
BACKGROUND
Technical Field
The present invention relates generally to the field of pharmaceutical
formulations for intravenous administration of vein irritating drugs.
Specifically, the
present invention relates to formulations for delivering vein irritating
antibiotic
drugs, such as vancomycin and clarithromycin.
Description of the Related Art
It is well known that many intravenously injected drugs cause vein
irritation or phlebitis at the injection site. Vancomycin is one of them.
Vancomycin
is an important antibiotic drug that is active against most gram-positive
bacteria,
and is particularly effective in clinically precarious settings, including
against
methicillin-resistent Staphyloceccus aureus. and other multidrug-resistant
organisms, against staphylococcal and streptococcal infections in penicillin-
and
cephalosporin-allergic patients, and against Clostridum difficile in
pseudomembranous colitis. Vancomycin is not absorbed well when given orally,
and intravenous injection is the main route of administration used for
systemic
infections. It is widely reported that vancomycin causes phlebitis, which is a
severe form of vein irritation and inflammation (Clin Pharm. 1988 Oct;
7(10):720-
1), with incidence occurred in up to 13% of patients (Archives of Pathology
and
Laboratory Medicine, 2000 Dec; 124(2)322-3). The prevention and treatment of
vancomycin associated phlebitis add significantly, as much as 45%, to the
total
cost of vancomycin treatment (Pharmacotherapy, 1994 Jul-Aug; 14(4):438-45).
The current intravenous vancomycin products are available in two
formulations. The first formulation is listed in the United State
Pharmacopoeia
(USP) as Sterile Vancomycin Hydrochloride, USP. It is provided in vials
containing
lyophilized (freeze-dried) sterile powder of vancomycin hydrochloride
equivalent to
either 500 mg or 1 g vancomycin activity. The formulation may also contain
hydrochloric acid and/or sodium hydroxide for pH adjustment. When
reconstituted
with Sterile Water for Injection, USP, it forms a clear solution with pH of
about 4.0
(2.5 to 4.5). The second formulation is provided as a frozen, iso-osmotic
sterile,
non-pyrogenic premixed 100 mL or 200 mL solution containing 500 mg or 1 g
Vancomycin, USP, respectively, as vancomycin hydrochloride. Each 100 mL of
solution contains approximately 5 g of Dextrose Hydrous, USP. The pH of the
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solution has been adjusted with hydrochloric acid and/or sodium hydroxide.
Thawed solutions have a pH in the range of 3.0 to 5Ø After thawing to room
temperature, this solution is available for intravenous use. This formulation
is
packaged in a plastic container (e.g., VANCOCIN HCI in GALAXY Plastic
Container (PL2040) for intravenous use Only), also listed in the USP..
Since vancomycin is sensitive to oxidation and hydrolysis
degradation in solution, it must be either lyophilized or frozen to provide
sufficiently
long shelf life. In solution, it is more stable in acidic than neutral or
alkaline
environments, therefore both the lyophilized and the frozen formulations are
adjusted to about pH 4.
While these simple solution formulations are able to preserve the
chemical integrity of vancomycin during storage, they are incapable of
preventing
the phlebitis caused by vancomycin. Vancomycin in its current formulations
must
be given slowly as a diluted solution, and the sites of infusion need be
rotated to
reduce frequency and severity of the vein irritation caused by vancomycin
(Product
Description for Sterile Vancomycin Hydrochloride, USP).
Clarithromycin is another vein irritating antibiotic when administered
intravenously. The currently available intravenous clarithromycin formulation,
KLARICID by Abbott Labs, is approved only in the United Kingdom and certain
other European countries, and is not licensed in the United States. Local
tolerability of intravenous clarithromycin formulation was reported as
problematic.
For example, Zimmerman et a1. (Clinical Drug Investigation 21: 527-36, 2001)
reported various adverse events at the injection sites from the intravenous
clarithromycin (KLARICID ) administration, including phlebitis (50%), vein
inflammation (75%), and vein irritation (100%).
BRIEF SUMMARY
Both freeze-stable and lyophilizable oil-in-water emulsions of
vancomycin are provided. In addition, frozen or lyophilized vancomycin
formulations that are stable upon storage and capable of being reconstituted
to
sub-micron size emulsions are also provided. Methods of preparing the oil-in-
water emulsions and frozen or lyophilized formulations of vancomycin are
further
provided. Similar methods may be used to prepare oil-in-water emulsions and
frozen or lyophilized formulations of clarithromycin. The resulting vancomycin
and
clarithromycin formulations may be used in treating or reducing infection and
other
diseases.
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In one aspect, a freeze-stable oil-in-water emulsion is provided that
comprises: (i) vancomycin at a concentration of about 0.1 % to about 3% by
weight
or a pharmaceutically acceptable salt or analog of vancomycin at an equivalent
concentration; (ii) one or more liquid oils at a total concentration of about
2% to
about 4% by weight; (iii) one or more phospholipids at a total concentration
of
about 2% to about 5% by weight, wherein the weight ratio of the total
phospholipid(s) to the total liquid oil(s) is at least 0.5:1; (iv) about 5% to
about 6%
dextrose by weight; and (v) water.
In certain embodiments, the one or more liquid oils of the freeze-
stable vancomycin oil-in-water emulsion comprise soybean oil.
In certain embodiments, the one or more liquid oils of the freeze-
stable oil-in-water vancomycin emulsion comprise a vegetable oil and a medium
chain triglyceride.
In certain embodiments, the one or more phospholipids of the freeze-
stable vancomycin oil-in-water emulsion comprise lecithin.
In certain embodiments, the freeze-stable oil-in-water emulsion
comprises: (1) vancomycin hydrochloride at a concentration of about 0.5% by
weight, (2) soybean oil at a concentration of about 1% to about 2% by weight,
(3)
medium chain triglyceride at a concentration of about 1% to about 2% by
weight,
(4) lecithin at a concentration of about 2% to about 4% by weight, and (5)
dextrose
at a concentration of about 5% to about 6% by weight.
In certain embodiments, the freeze-stable oil-in-water emulsion
comprises: (1) vancomycin hydrochloride at a concentration of about 0.5% by
weight, (2) soybean oil and medium chain triglyceride, wherein the total
concentration of soybean oil and medium chain triglyceride is about 2% by
weight,
the weight ratio of soybean oil to medium chain triglyceride is between 2:1 to
1:1,
(3) lecithin at a concentration of about 2% by weight, and (4) dextrose at a
concentration of about 5% by weight.
In certain embodiments, the average size of the oil droplets in the
freeze-stable vancomycin oil-in-water emulsion is no more than about 200 nm,
and
the PFAT5 is less than about 0.05.
In certain embodiments, the freeze-stable oil-in-water vancomycin
emulsion does not further comprise a compound that increases the amount of
vancomycin in the oil droplets of the emulsion
In certain embodiments, less than about 30% of vancomycin is in the
oil droplets of the freeze-stable oil-in-water emulsion.
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In another aspect, a frozen composition is provided that comprises:
(i) vancomycin at a concentration of about 0.5% by weight or a
pharmaceutically
acceptable salt or analog of vancomycin at an equivalent concentration; (ii)
one or
more liquid oils at a total concentration of about 2% to about 4% by weight;
(iii) one
or more phospholipids at a total concentration of about 1% to about 5% by
weight;
wherein the weight ratio of the total phospholipid(s) to the total liquid
oil(s) is at
least 0.5:1; (iv) about 5% to about 6% dextrose by weight; and (v) water;
wherein
when thawed, the composition forms an oil-in-water emulsion with an average
diameter of oil droplets no more than about 200 nm and a PFAT of less than
about
0.05.
In certain embodiments, the one or more liquid oils of the frozen
vancomycin composition comprise a vegetable oil and a medium chain
triglyceride.
In certain embodiments, the frozen vancomycin composition does not
further comprise a compound that increases the amount of vancomycin in the oil
droplets of the emulsion formed by thawing the frozen vancomycin composition.
In certain embodiments, when the frozen vancomycin composition is
thawed to form an oil-in-water emulsion, less than about 30% of vancomycin is
present in the oil droplets of the oil-in-water emulsion.
In another aspect, a method for treating or reducing infection is
provided that comprises administering to a patient in need thereof a
pharmaceutically effective amount of the freeze-stable vancomycin oil-in-water
emulsion described herein.
In a related aspect, a method for treating or reducing infection is
provided that comprises administering to a patient in need thereof a
pharmaceutically effective amount of an oil-in-water vancomycin emulsion
formed
by thawing the frozen vancomycin composition described herein.
In another aspect, a freeze-stable oil-in-water emulsion is provided
that comprises: (i) clarithromycin at a concentration of at least about 0.5%
by
weight or a pharmaceutically acceptable salt or ester of clarithromycin at an
equivalent concentration; (ii) one or more liquid oils at a total
concentration of
about 2% to about 4% by weight; (iii) one or more phospholipids at a total
concentration of about 1% to about 5% by weight, wherein the weight ratio of
the
total phospholipid(s) to the total liquid oil(s) is at least 0.5:1; (iv) about
5% to about
6% dextrose by weight; and (v) water.
In certain embodiments, the one or more liquid oils of the freeze-
stable clarithromycin oil-in-water emulsion comprise soybean oil.
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In certain embodiments, the one or more liquid oils of the freeze-
stable clarithromycin oil-in-water emulsion comprise a vegetable oil and a
medium
chain triglyceride.
In certain embodiments, the one or more phospholipids of the freeze-
stable clarithromycin oil-in-water emulsion comprise lecithin.
In certain embodiments, the freeze-stable clarithromycin oil-in-water
emulsion does not further comprise a compound that increases the amount of
clarithromycin in the oil droplets of the emulsion.
In certain embodiments, less than about 30% of clarithromycin is in
the oil droplets of the freeze-stable clarithromycin oil-in-water emulsion.
In certain embodiments, the freeze-stable oil-in-water emulsion
comprises: (1) clarithromycin at a concentration of about 1% to about 5% by
weight, (2) soybean oil and medium chain triglyceride, wherein the total
concentration of soybean oil and medium chain triglyceride is about 2% by
weight,
the weight ratio of soybean oil to medium chain triglyceride is between 2:1 to
1:1;
(3) lecithin at a concentration of about 2% by weight; and (4) dextrose at a
concentration of about 5% by weight.
In certain embodiments, the average size of the oil droplets in the
freeze-stable oil-in-water clarithromycin emulsion is no more than about 200
nm,
and the PFAT5 is less than about 0.05.
In another aspect, a frozen composition is provided that comprises:
(i) clarithromycin at a concentration of at least about 0.5% by weight or a
pharmaceutically acceptable salt or ester of clarithromycin at an equivalent
concentration; (ii) one or more liquid oils at a total concentration of about
2% to
about 4% by weight; (iii) one or more phospholipids at a total concentration
of
about 1% to about 5% by weight, wherein the weight ratio of the total
phospholipid(s) to the total liquid oil(s) is at least 0.5:1; (iv) about 5% to
about 6%
dextrose by weight; and (v) water; wherein when thawed, the composition forms
an
oil-in-water emulsion with an average diameter of oil droplets no more than
about
200 nm and a PFAT5 of less than 0.05.
In certain embodiments, the one or more liquid oils of the frozen
clarithromycin composition comprise a vegetable oil and a medium chain
triglyceride.
In another aspect, a method for treating or reducing infection is
provided that comprises administering to a patient in need thereof a
pharmaceutically effective amount of the freeze-stable oil-in-water
clarithromycin
emulsion as described herein.
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In another aspect, a method for treating or reducing infection
comprising administering to a patient in need thereof a pharmaceutically
effective
amount of an oil-in-water clarithromycin emulsion formed by thawing the frozen
clarithromycin composition as described herein.
In one aspect, the present invention provides a lyophilizable oil-in-
water emulsion comprising: (i) at least about 15 mg/mI vancomycin or a
pharmaceutically acceptable salt or analog thereof, (ii) one or more liquid
oils at a
total concentration of about 2% to about 10% by weight, (iii) one or more
phospholipids at a total concentration of about 1% to about 10% by weight, and
(iv)
dextrose at a concentration of at least about 10% by weight.
In certain embodiments, the one or more liquid oils of the
lyophilizable vancomycin oil-in-water emulsion comprise soybean oil.
In certain embodiments, the one or more phospholipids of the
lyophilizable vancomycin oil-in-water emulsion comprise lecithin.
In certain embodiments, the lyophilizable oil-in-water emulsion
comprises vancomycin hydrochloride at a concentration of about 1% to about 3%
by weight, medium chain triglyceride at a concentration of about 1% to about
5%
by weight, vegetable oil at a concentration of about 1% to about 5% by weight,
lecithin at a concentration of about 1% to about 4% by weight, and dextrose of
about 15% to about 25% by weight.
In certain embodiments, the pH of the lyophilizable vancomycin oil-in-
water emulsion is about 3 to about 8.
In certain embodiments, the average size of the oil droplets in the
lyophilizable vancomycin oil-in-water emulsion is less than about 250 nm.
In certain embodiments, the average size of the oil droplets in the
lyophilizable vancomycin oil-in-water emulsion is less than about 200 nm and
the
PFAT5 of the emulsion is less than about 0.05.
In certain embodiments, the lyophilizable vancomycin oil-in-water
emulsion does not further comprise a compound that increases the amount of
vancomycin in the oil droplets of the emulsion.
In certain embodiments, less than about 30% of vancomycin is
present in the oil droplets of the lyophilizable vancomycin oil-in water
emulsion.
In another aspect, the present invention provides a lyophilized
composition comprising vancomycin or a pharmaceutically acceptable salt or
analog thereof, liquid oil(s), phospholipid(s), and dextrose, wherein the
composition is prepared by removal of water from the oil-in-water emulsion
described herein, the composition can be rehydrated with water to form an
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emulsion suitable for injection, and the average diameter of the re-formed
emulsion droplets is no greater than about 1 micron.
In certain embodiments, the average diameter of the reformed
emulsion droplets is no greater than about 400 nm.
In another aspect, the present invention provides a lyophilized
composition comprising: (i) vancomycin or a pharmaceutically acceptable salt
or
analog thereof at a concentration of about 5% to about 10% by weight, (ii) one
or
more liquid oils at a total concentration of about 10% to about 20% by weight,
(iii)
one or more phospholipids at a total concentration of about 10% to about 20%
by
weight, and (iv) dextrose at a concentration 50% to about 80% by weight.
In certain embodiments, the lyophilized vancomycin composition
forms an oil-in-water emulsion when rehydrated with water, and the average
diameter of the emulsion droplets is no greater than about 1 micron.
In certain embodiments, the lyophilized vancomycin composition
does not further comprise a compound that increases the amount of vancomycin
in
the oil droplets of the emulsion reconstituted from the lyophilized vancomycin
composition.
In certain embodiments, when the lyophilized vancomycin
composition is rehydrated with water to form an oil-in-water emulsion, less
than
about 30% of vancomycin is present in the oil droplets of the oil-in-water
emulsion.
In certain embodiments, the composition forms an oil-in-water
emulsion when rehydrated with water, the average size of the oil droplets in
the
emulsion is no more than about 200 nm, and the PFAT5 of the emulsion is less
than about 0.05.
In another aspect, the present invention provides a method for
treating or reducing the risk of infection comprising administrating to a
patient in
need thereof a pharmaceutically effective amount of the lyophi(izabfe oil-in-
water
emulsion of vancomycin described herein.
In a related aspect, the present invention provides a method for
treating or reducing the risk of infection comprising administering to a
patient in
need thereof a pharmaceutically effective amount of an oil-in-water emulsion
formed by rehydrating the lyophilized vancomycin composition described herein.
In another aspect, the present invention provides a lyophilizable oil-
in-water emulsion comprising: (i) clarithromycin or a pharmaceutically
acceptable
salt or ester thereof at a concentration of at least about 1% by weight, (ii)
one or
more liquid oils at a total concentration of about 2% to about 10% by weight,
(iii)
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one or more phospholipids at a total concentration of about 1% to about 10% by
weight, and (iv) dextrose at a concentration of at least about 10% by weight.
In certain embodiments, the lyophilizable oil-in-water emulsion
comprises clarithromycin or its pharmaceutically acceptable salt or ester at a
concentration of about 1% to about 3% by weight, medium chain triglyceride at
a
concentration of about 1% to about 5% by weight, vegetable oil at a
concentration
of about 1 /a to about 5% by weight, lecithin at a concentration of about 1%
to 10%
by weight, and dextrose of about 15% to 25% by weight.
In certain embodiments of the lyophilizable oil-in-water emulsion of
clarithromycin, the average size of the oil droplets in the emulsion is less
than
about 250 nm.
In certain embodiments of the lyophilizable oil-in-water emulsion of
ctarithromycin, the average size of the oil droplets in the emulsion is less
than
about 200 nm, and the PFAT5 of the emulsion is less than about 0.05.
In certain embodiments of the lyophilizable oil-in-water emulsion of
clarithromycin, the emulsion does not further comprise a compound that
increases
the amount of clarithromycin in the oil droplets of the emulsion.
In certain embodiments of the lyophilizable oil-in-water emulsion of
clarithromycin, less than about 30% of clarithromycin is in the oil droplets
of the
emulsion.
In another aspect, the present invention provides a lyophilized
composition comprising clarithromycin or a pharmaceutically acceptable salt or
ester thereof, liquid oil(s), phospholipid(s), and dextrose, wherein the
composition
is prepared by removal of water from the oil-in-water emulsion of
clarithromycin
described herein, the composition can be rehydrated with water to form an
emulsion suitable for injection, and the average diameter of the re-formed
emulsion droplets is no greater than about 1 micron.
In certain embodiments, the average diameter of oil droplets in the
emulsion reconstituted from the lyophilized clarithromycin composition is no
greater than about 200 nm.
In a related aspect, the present invention provides a lyophilized
composition comprising: (i) clarithromycin or a pharmaceutically acceptable
salt or
ester thereof at a concentration of about 2% to about 8% by weight, (ii)
liquid oil at
a concentration of about 10% to about 20% by weight, (iii) one or more
phospholipids at a concentration of about 10% to about 20% by weight, and (iv)
dextrose at a concentration about 50% to about 80% by weight.
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In certain embodiments, the lyophilized clarithromycin composition as
described herein forms an oil-in-water emulsion when rehydrated with water,
and
the average diameter of the emulsion droplets is no greater than about 200 nm.
In certain embodiments, the lyophitized clarithromycin composition as
described herein forms an oil-in-water emulsion when rehydrated with water,
and
the PFAT of the emulsion is less than about 0.05.
In certain embodiments, the lyophilized clarithromycin composition as
described herein forms an oil-in-water emulsion when rehydrated with water,
the
average diameter of the emulsion droplets is no greater than about 200 nm, and
the PFAT of the emulsion is less than about 0.05.
In certain embodiments, the lyophilized clarithromycin composition
does not further comprise a compound that increases the amount of
clarithromycin
in the oil droplets of the emulsion reconstituted from lyophilized
composition.
In certain embodiments, the lyophilized clarithromycin composition as
described herein forms an oil-in-water emulsion when rehydrated with water,
and
less than about 30% of clarithromycin is in the oil droplets of the emulsion.
In another aspect, the present invention provides a method for
treating or reducing the risk of infection comprising administrating to a
patient in
need thereof a pharmaceutically effective amount of the lyophilizable oil-in-
water
emulsion of clarithromycin described herein.
In a related aspect, the present invention provides a method for
treating or reducing the risk of infection comprising administering to a
patient in
need thereof a pharmaceutically effective amount of an oil-in-water emulsion
formed by rehydrating the lyophilized composition of clarithromycin described
herein.
DETAILED DESCRIPTION
Both freeze-stable and lyophilizable oil-in-water emulsions are
provided for the vein irritating antibiotics vancomycin and clarithromycin.
Methods
for preparing the oil-in-water emulsions and frozen or lyophilized
formulations are
also provided. Methods for using the oil-in-water emulsions directly prepared
or
reconstituted from frozen or lyophilized formulations in treating or reducing
infection are further provided.
A term used in one subsection of this specification has the same
meaning in another subsection unless otherwise noted.
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Freeze-stable Oil-in-Water Emulsions and Frozen Formulations
In one aspect, freeze-stable oil-in-water emulsions are provided for
vancomycin and clarithromycin. Such emulsions may have one or more of the
following advantages: (1) they may be frozen for storage and remain stable
during
storage, (2) they contain sub-micron oil droplets and may be filter
sterilized, (3) the
frozen emulsions may be thawed to re-form oil-in-water emulsions with sub-
micron
oil droplets, and the re-formed emulsions may also be filter sterilized, (4)
the
emulsions that are either directly prepared or re-formed from the frozen
emulsions
are isotonic and ready for administration to a patient in need of such a
treatment,
(5) the oil-in-water emulsions that are either directly prepared or re-formed
from
the frozen emulsions are non-vein irritating; and (6) the oil-in-water
emulsions do
not contain any additional compounds (e.g., stabilizers) that increase the
amount
of vancomycin or clarithromycin in the oil droplets, which avoids any side
effects of
these additional compounds.
Vancomycin Freeze-Stable and Frozen Formulations
Vancomycin is a tricyclic glycopeptide (whose structure is shown
below). In an aqueous solution, it has limited stability and is most stable at
pH
about 4.0-4.5; so as to maximize its stability, the currently marketed
formulation
(Sterile Vancomycin Hydrochloride, USP) is formulated at pH 4.0 to 4.5 in a
lyophilized form. Vancomycin is moderately soluble in water (to about 1% to
about
5%). It is more soluble in an acidic or basic than a pH neutral environment.
Its
solubility reaches a minimum at about pH 7. As the pH changes from pH 4.0-4.5
(i.e., the formulation pH) to pH 7 (i.e., blood pH), the solubility of
vancomycin is
reduced by about 40%. Thus, there is a potential for a partial drug
precipitation in
the vein upon an intravenous injection where the acidic formulation solution
is
mixed with the pH neutral blood. Such a partial drug precipitation might be a
factor
that contributes to the vein irritation caused by the currently marketed
formulation.
CA 02651988 2008-11-12
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HO NH2
CHb C)H
H3C
p 0 OH
0
Ca Ci
c1 I1 LLN 4 ~
C7 0 0
O .,,N~.., ~!~, . NHCH3
HN I~1 H
H2N
HOOC
~ ~= ~ 0
HO OHOH
The oil-in-water emulsions provided herein are a safe and effective
formulation option for intravenous delivery of vancomycin. Although not
wishing to
be bound, it is hypothesized that the oil droplets of vancomycin oil-in-water
emulsions prevent or reduce vein irritation of vancomycin by (1) interfering
with the
binding of vancomycin to the endothelium of the vein, (2) confining a portion
of
vancomycin molecules in the emulsion oil droplets, thus reducing direct
contact
with vein endothelium, and/or (3) improving the vancomycin solubility by
allowing
vancomycin to dissolve in the oil droplets and reducing the possibility of
vancomycin precipitation in the blood stream and depositing or binding to the
vein
at the injection site. `
In one aspect, a freeze-stable oil-in-water emulsion is provided that
comprises: (i) vancomycin at a concentration of about 0.1 % to about 3% by
weight
or a pharmaceutically acceptable salt or analog of vancomycin at an equivalent
concentration; (ii) one or more liquid oils at a total concentration of about
2% to
about 4% by weight; (iii) one or more phospholipids at a total concentration
of
about 2% to about 5% by weight, wherein the weight ratio of the total
phospholipid(s) to the total liquid oil(s) is at least 0.5:1; (iv) about 5% to
about 6%
dextrose by weight; and (v) water.
An "oil-in-water emulsion," as used herein, refers to a colloidal
dispersion system in which liquid oil is dispersed in small droplets (the
discrete
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phase, also referred to as "the oil phase") in an aqueous medium (the
continuous
phase, also referred to as "the aqueous phase").
"Pharmaceutically acceptable salts or analogs," as used herein,
refers to salts or structurally related chemicals, which are, within the scope
of
sound medical judgment, suitable for use in contact with the tissues of humans
and lower animals without undue toxicity, irritation, allergic response, and
the like,
and effective for their intended use in treating or preventing infections.
Pharmaceutically acceptable salts of vancomycin include hydrochloride,
sulfate,
mesylate, acetate, citrate, ascorbate, aspartate, besylate, benzoate,
decanoate,
hexanoate, octanoate, cyclamate, laurylsulfate, formate, fumarate,
glucoheptonate,
glucuronate, glutamate, glutarate, glycolate, hydrobromate, lactate,
lactobionate,
maleate, malonate, nicotinate, nitrate, oleate, orotate, oxalate, palmitate,
pamoate,
phosphate, stearate, succinate, tartarate, etc. Pharmaceutically acceptable
analogs of vancomycin refer to glycopeptide chemicals having a structure and
biological activities similar to vancomycine. Examples of such analogs include
LY333328 and BI 397 (Malabarba A, Ciabatti R. Glycopeptide derivatives. Curr
Med Chem. 2001; 8:1759-1773), Teicoplanin (formerly known as teichmycin A2(1),
Salaria M. Teicoplanin. Indian Pediatr. 2001;38:372-375). Telavancin (TLV)
(Douchin K, Shaw J, Spencer E, Seroogy J, Barriere S, Wilbraham D. Single dose
pharmacokinetics (PK) of telavancin (TLV) in healthy elderly subjects. Clin
Microbio! Infect. 2004;10:275), Oritavancin (Internet press release from
Biospace
Beat. Biotechnology and Pharmaceutical News), Targocid
(http://home.intekom.com/pharm/roussel/targocid.html), and norvancomycin.
"Concentration by weight," as used herein, refers to the ratio (in
percentage) of the weight of a component (e.g., vancomycin) of a composition
(e.g., a vancomycin oil-in-water emulsion) to the total weight of the
composition.
A pharmaceutically acceptable salt of vancomycin is at a
concentration "equivalent" to a specified concentration of vancomycin if at
that
concentration, the vancomycin salt contains the same amount of vancomycin free
base as vancomycin at the specified concentration. For example, 5.16 mg/g
vancomycin hydrochloride is equivalent to 5.0 mg/g vancomycin free base.
A-pharmaceutically acceptable analog of vancomycin is at a
concentration "equivalent" to a specified concentration of vancomycin if at
that
concentration, the vancomycin analog is as effective as vancomycin at the
specified concentration in treating or preventing infection.
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In the freeze-stable oil-in-water emulsions provided herein,
vancomycin is present at a concentration of about 0.1 % to about 3% by weight,
including any value from 0.09% to 3.3%.
The term "about," as used in the present disclosure, refers to any
value in the range of 90% to 110% of a specified value. For example, about
0.1%
refers to any percentage from 0.09% to 0.11 %.
The term "liquid oil" is used in the present disclosure in a general
sense to identify hydrocarbon derivatives, carbohydrate derivatives, or
similar
organic compounds that are liquid at body temperatures, i.e., about 37 C, and
are
pharmacologically acceptable in injectable formulations. This class includes
vegetable oils, animal fats, and synthetic oils, as well as various liquids
that are
obtained by chemical treatment of such oils and fats.
The term "oil component" refers to an oil, or a combination of multiple
oils in an oil-in-water emulsion.
In certain embodiments, the oil component of oil-in-water emulsions
provided herein comprises a monoglyceride, a diglyceride, a triglyceride, or a
mixture thereof. In certain embodiments, the oil component comprises an ester
formed between one or more fatty acids and an alcohol other than glycerol.
"Vegetable oil" refers to oil derived from plant seeds or nuts.
Exemplary vegetable oils include, but are not limited to, almond oil, borage
oil,
black currant seed oil, corn oil, safflower oil, soybean oil, coitonseed oil,
peanut oil,
olive oil, rapeseed oil, coconut oil, palm oil, canola oil, etc. In certain
embodiments, the one or more liquid oils of the oil-in-water emulsion comprise
soybean oil.
Vegetable oils are typically "long-chain triglycerides," formed when
three fatty acids (usually about 14 to about 22 carbons in length, with
unsaturated
bonds in varying numbers and locations, depending on the source of the oil)
form
ester bonds with the three hydroxyl groups on glycerol. In certain
embodiments,
vegetable oils of highly purified grade (also called "super refined") are
generally
used to ensure safety and stability of oil-in-water emulsions.
"Medium chain triglycerides" (MCT's) is another class of triglyceride
oil that can be either naturally derived or synthetic. MCT's are made from
fatty
acids that are usually about 6 to about 12 carbons in length. Like vegetable
oils,
MCT's have been used extensively in emulsions designed for injection as a
source
of calories, for patients requiring parenteral nutrition. Such oil is
commercially
available as Miglyol 812 from SASOL GmbH, Germany, CRODAMOL GTCC-PN
from Croda Inc. of Parsippany, New Jersey, or Neobees M-5 oil from PVO
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International, Inc., of Boonton, New Jersey. Other low-melting medium chain
oils
may also be used in the emulsions provided herein.
"Animal fat" refers to oil derived from an animal source. It also
comprises triglycerides, but the lengths of, and unsaturated bonds in, the
three
fatty acid chains vary, compared to vegetable oils. Animal fats from sources
that
are solid at room temperature (such as tallow, lard, etc.) can be processed to
render them liquid if desired. Other types of animal fats that are inherently
liquid at
room temperature include various fish oils, etc.
The freeze-stable oil-in-water emulsions provided herein contain one
or more liquid oils at a total concentration of about 2% to about 4% by
weight,
including any values from 1.8% to 4.4%.
In certain embodiments, combinations of vegetable oil and MCT oil
are used in the emulsions provided herein. Such combinations generally have a
long record of safe use in combination in injectable emulsions and provide
superior
stability for the emulsions of this invention. The specific type of vegetable
oil used
(i.e., soy bean oil, corn oil, or safflower oil, etc.) is not critical, so
long as it is safe,
well tolerated, pharmaceutically acceptable, chemically stable, and provides
emulsion droplets having a desired size range. Generally, MCT oil is limited
to at
most 50% by weight in the combinations of vegetable oil and MCT.
A "phospholipid," as used herein, refers to a triester of glycerol with
two fatty acids and one phosphate ion. Exemplary phospholipids useful in the
emulsions provided herein include, but are not limited to, phosphatidyl
chlorine,
lecithin (a mixture of choline ester of phosphorylated diacylglyceride),
phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid with about 4
to
about 22 carbon atoms, and more generally from about 10 to about 18 carbon
atoms and varying degrees of saturation. The phospholipid component of the
emulsions of the present inventiori can be either a single phospholipid or a
mixture
of several phospholipids.
The phospholipids useful in the emulsions provided herein can be of
natural origin. Naturally occurring lecithin is a mixture of the diglycerides
of stearic,
palmitic, and oleic acids, linked to the choline ester of phosphoric acid,
commonly
called phosphatidylcholine, and can be obtained from a variety of sources such
as
eggs and soy beans. Soy lecithin and egg lecithin (including hydrogenated
versions of these compounds) have a long history of safety, possess combined
emulsification and solubilization properties, and tend to be broken down into
innocuous substances more rapidly than most synthetic surfactants.
Commercially
available soya phospholipids are the Centrophase and Centrolex products
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marketed and sold by Central Soya, Phospholipon from Phospholipid GmbH,
Germany, Lipoid by Lipoid GmbH, Germany, and EPIKURON by Degussa.
Phospholipids useful in the present invention can also be
synthesized. Exemplary common synthetic phospholipids are listed below:
Diacylglyicerols
1,2-Dilauroyl-sn-glycerol (DLG)
1,2-Dimyristoyi-sn-glycerol (DMG)
1,2-Dipalmitoyl-sn-glycerol (DPG)
1,2-Distearoyl-sn-glycerol (DSG)
Phosphatidic Acids
1,2-Dimyristoyl- sn-glycero-3-phosphatidic acid, sodium salt (DMPA,Na)
1,2-Dipalmitoyl- sn-glycero-3-phosphatidic acid, sodium salt (DPPA,Na)
1,2-Distearoyl- sn-glycero-3-phosphatidic acid, sodium salt (DSPA,Na)
Phosphocholines
1,2-Dilauroyl-sn-glycero-3-phosphocholine (DLPC)
1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC)
1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)
1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)
1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC)
1,2-Distearoyl-sn-glycero-3-phosphochofine (DSPC)
Phosphoethanolamines
1,2-Dilauroyl-sn-glycero-3-phosphoethanolamine (DLPE)
1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE)
1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE)
1,2-Distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)
Phosphoglycerols
1,2-Dilauroyl-sn-glycero-3-phosphoglycerol, sodium salt (DLPG)
1,2-Dimyristoyl-sn-glycero-3-phosphoglycerol, sodium salt (DMPG)
1,2-Dimyristoyl-sn-gfycero-3-phospho- sn-l-glycerol, ammonium salt
(DMP-sn-1-G,NH4)
1,2-Dipalmitoyl- sn-glycero-3-phosphoglycerol, sodium salt (DPPG,Na)
1,2-Distearoyl- sn-glycero-3-phosphoglycerol, sodium salt (DSPG,Na)
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1,2-Distearoyl- sn-glycero-3-phospho- sn-l-glycerol, sodium salt (DSP-sn-1 G,
Na)
Phosphoserines
1,2-Dipalmitoyl- sn-glycero-3-phospho-L-serine, sodium salt (DPPS,Na)
Mixed Chain Phospholipids
1-Palmitoyl-2-oleoyl- sn-glycero-3-phosphocholine (POPC)
1-Paimitoyl-2-oleoyl- sn-glycero-3-phosphoglycerol, sodium salt (POPG,Na)
1-Palmitoyl-2-oleoyl- sn-glycero-3-phosphoglycerol, ammonium salt (POPG,NH4)
Lysophosgholipids
1-Paimitoyl-2-lyso- sn-glycero-3-phosphocholine (P-lyso-PC)
1-Stearoyl-2-lyso- sn-g lycero-3-phosphocho line (S-lyso-PC)
Peqylated Phospholipids
N-(Carbonyi-methoxypolyethyleneglycoi 2000)- MPEG-2000-DPPE
1,2-dipalmitoyl- sn-glycero-3-phosphoethanolamine, sodium salt
N-(Carbonyl-methoxypolyethyleneglycol 5000)- MPEG-5000-DSPE
1,2-distearoyl- sn-glycero-3-phosphoethanolamine, sodium salt
N-(Carbonyl-methoxypolyethyleneglycol 5000)- MPEG-5000-DPPE
1,2-dipalmitoyl- sn-glycero-3-phosphoethanolamine, sodium salt
N-(Carbonyl-methoxypolyethyleneglycol 750)- MPEG-750-DSPE
1,2-distearoyl- sn-glycero-3-phosphoethanolamine, sodium salt
N-(Carbonyl-methoxypolyethyleneglycol 2000)- MPEG-2000-DSPE
1,2-distearoyl- sn-glycero-3-phosphoethanolamine, sodium salt
In certain embodiments, the one or more phospholipids of the freeze-
stable oil-in-water emulsion comprise lecithin, such as soy lecithin or egg
lecithin.
The freeze-stable oil-in-water emulsion provided herein comprises
one or more phospholipids at a total concentration of about 2% to about 5% by
weight (including any value between 1.8% and 5.5%).
The weight ratio of the total phospholipid(s) to the total liquid oil(s) in
the freeze-stable oil-in-water emulsion provided herein is at least 0.5:1. In
certain
embodiments, the weight ratio of the total phospholipid(s) to the total liquid
oil(s) is
1:1.
The freeze-stable oil-in-water emulsion provided herein comprises
about 5% to about 6% dextrose by weight.
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In most embodiments, the freeze-stable oil-in-water emulsion
provided herein is "isotonic" (used interchangeably with "iso-osmotic"). An
oil-in-
water emulsion is isotonic when its measured osmotic pressure equals to that
of
normal saline (0.9% sodium chloride), which is about 296 mOsm. Typical
measurement variability for an emulsion sample is about 10%; therefore, an
emulsion with a measured osmotic pressure between 266 and 326 mOsm is
regarded as iso-osmotic or isotonic.
In certain embodiments, the oil droplets of the freeze-stable oil-in-
water emulsions are of sub-micron size. A "sub-micron size droplet," as used
herein, refers to an oil droplet in an oil-in-water emulsion having an average
diameter of less than 1 micron as measured by conventional sizing techniques
such as laser light scattering spectrometry. Oil droplets of sub-micron size
are
desired for the safe passage of these droplets in capillary blood vessel
circulation.
Droplets of greater than 5 micron in diameter are believed to be unsafe for
intravenous injection since they may block capillary blood vessels resulting
in
pulmonary embolism.
In certain embodiments, the oil droplets of the freeze-stable oil-in-
water emulsions have an average diameter of less than 500, 450, 400, 350, 300,
or 250 nm.
In certain embodiments, the oil droplets of the freeze-stable
formulations have an average diameter of less than 0.2 micron (200 nm) so that
the emulsion may be sterilized by filtering through a 0.2 micron rated filter
membrane. In certain embodiments, the oil droplets of the compositions of the
present invention have an average diameter of less than about 175, 150, 125,
100,
or 75 nm.
In certain embodiments, the freeze-stable oil-in-wateremulsions also
have a PFAT5 value of less than 0.05. "PFAT5" refers to the volume percent of
fat
or oil droplets in diameter greater than 5 pm. A PFAT5 of less than 0.05% is
preferred for an intravenous emulsion (Todd Canada, "Pathological
Consequences From the Infusion of Unstable Lipid Emulsion Admixtures in Guinea
Pigs" Nutrition in Clinical Practice, Vol. 21, No. 6, 2006 636-637). .A PFAT5
value
can be measured by the method of single-particle optical sensing (SPOS), also
called optical particle counting (OPC) such as AccuSizer 780 by Particle
Sizing
System.
In certain embodiments, the freeze-stable oil-in-water emulsion has
an average size of the oil droplets of no more than about 200 nm and a PFAT5
less than about 0.05. The low PFAT5 reduces or eliminates the potential for
large
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oil droplets to block the capillary circulation. The small average size of the
oil
droplets allows the emulsion be filter sterilized, which is particularly
preferred for
vancomycin due to its heat, light and oxygen sensitivities. Such sensitivities
render
other sterilization methods, such as autoclave, gamma radiation, or UV
exposure,
infeasible or less preferred.
An oil-in-water emulsion is "freeze-stable" if after one cycle of freeze
(at -20 C) - thaw (at 4 C or 25 C), the average diameter of the oil droplets
in the
emulsion does not increase by more than 50%.
In certain embodiments, the freeze-stable oil-in-water emulsions
provided herein do not increase their average diameters of the oil droplets by
more
than 50% after 2, 3, 4, or 5 freeze-thaw cycles.
In certain embodiments, the average diameters of the oil droplets of
the freeze-stable oil-in-water emulsions provided herein do not increase by
more
than 40%, 30%, 30%, 20%, or 10% after 1, 2, 3, 4, or 5 freeze-thaw cycles.
In certain embodiments, the freeze-stable oil-in-water emulsion
retains its average oil droplet size of no more than about 200 nm and a PFAT5
of
less than about 0.05 after 1, 2, 3, 4, or 5 freeze-thaw cycles.
The freeze-stable oil-in-water emulsions disclosed herein are stable
when stored at a frozen state (e.g., at -20 C) for an extended period of time
(e.g.,
for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months) as described in more
detail below.
The freeze-stable oil-in-water emulsions disclosed herein when stored at a
frozen
state are also referred to as "frozen compositions," "frozen emulsions," or
"frozen
formulations."
In certain embodiments, the pH of the freeze-stable oil-in-water
emulsions of vancomycin is about 3 to about 8. In certain embodiments, the pH
is
about 4 to about 7.
In certain embodiments, the freeze-stable oil-in-water emulsion does
not further comprise a compound that increases the amount of vancomycin in the
oil droplets of the oil-in-water emulsion. Examples of such compounds include
fatty acids, cholesterol sulfate, riboflavin-5-phosphate, vitamin E succinate,
and a
mixture of two or more of the above-listed agents. The answer is "no"
according to
Andrew.
In certain other embodiments, although the freeze-stable oil-in-water
emulsion may further comprise a compound that increases the amount of
vancomycin in the oil droplets, the stability (physical and/or chemical
stability) of
the emulsion does not require the presence of such a compound.
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In certain embodiments, less than about 70% of vancomycin is in the
oil droplets of the freeze-stable oil-in-water emulsion. In certain
embodiments, less
than about 60%, 50%, 40%, 30%, or 20% of vancomycin is in the oil droplets of
the
freeze-stable oil-in-water emulsion.
In certain embodiments, some or all of the components other than
vancomycin in the freeze-stable oil-in-water emulsion (e.g., an oil component
and
a phospholipid) are safe, well tolerated, and acceptable by the FDA for
intravenous
injection.
A component of oil-in-water emulsions is regarded as "safe" if it does
not cause undesired systemic reactions such as anaphylactic shock in patients.
A component of oil-in-water emulsions is regarded as 'Well tolerated"
if it does not result in substantially adverse effects at the injection site,
such as
phlebitis, vein inflammation or vein irritation.
A component of oil-in-water emulsions is regarded as "acceptable by
the FDA" if it has been approved by the FDA for intravenous injection as of
the
filing date of the present application, and is being used at a concentration
comparable to those used in FDA approved products.
In certain embodiments, some or all of the components other than
the vancomycin in the freeze-stable oil-in-water emulsion (e.g., an oil
component
and a phospholipid) are generally regarded as safe for use in intravenous
injections by a drug regulatory authority.
A component of oil-in-water emulsions is "generally regarded as safe
for use in intravenous injections by a drug regulatory authority" if it has
been used
in intravenous injection products approved by the FDA or a drug regulatory
authority in Europe as of the filing date of the present application, and is
being
used at a concentration comparable to that used in the products approved by
the
FDA in the United States or by a drug regulatory authority in Europe.
In certain embodiments, the freeze-stable oil-in-water emulsions are
vein non-irritable. "Vein non-irritable" refers to the property of a compound
or
composition, when administered intravenously, that does not cause substantial
irritation at the injection site, as evident by, for example, thickened skin,
necrotic
skin, local redness, local swelling, venous dilation with blood clot
formation, or
venous embolism with subcutaneous inflammation. This term is used
interchangeable with "non-vein irritable", "non-vein irritating" or the like.
In certain embodiments, the freeze-stable oil-in-water emulsion or its
component(s) is injectable. "Injectable" refers to acceptance of an ingredient
by a
drug authority agent (e.g., the US FDA) by allowing it for use in an injection
drug.
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In certain embodiments, the freeze-stable oil-in-water emulsion or its
component(s) is biocompatible. "Biocompatible" refers to the capability of
performing functions within or upon a living organism in an acceptable manner,
i.e., without undue toxicity or harmful physiological or pharmacological
effects.
An exemplary freeze-stable oil-in-water emulsion of the present
invention comprises: (1) vancomycin hydrochloride at a concentration of about
0.5% by weight, (2) soybean oil at a concentration of about 9% to about 2% by
weight, (3) medium chain triglyceride at a concentration of about 1% to about
2%
by weight, (4) lecithin at a concentration of about 2% to about 4% by weight,
and
(5) dextrose at a concentration of about 5% to about 6% by weight.
Another exemplary freeze-stable oil-in-water emulsion of the present
invention comprises: (1) vancomycin hydrochloride at a concentration of about
0.5% by weight, (2) soybean oil and medium chain triglyceride, wherein the
total
concentration of soybean oil and medium chain triglyceride is about 2% by
weight,
the weight ratio of soybean oil to medium chain triglyceride is between 2:1 to
1:1,
(3) lecithin at a concentration of about 2% by weight, and (4) dextrose at a
concentration of about 5% by weight.
The present invention also provides methods for preparing freeze-
stable oil-in-water emulsions of vancomycin described herein. Such emulsion
compositions may be prepared by (a) forming a mixture that comprises (i) one
or
more liquid oils (e.g., a vegetable oil, or a combination of a vegetable oil
and a
medium chain triglyceride) and (ii) one or more phospholipids, (b) forming a
mixture that comprises (i) a pharmaceutically effective amount of vancomycin
or a
pharmaceutically acceptable salt or analog thereof, (ii) dextrose, and (iii)
water,
and (c) forming an oil-in-water emulsion with the mixtures of step (a) and
(b).
In certain embodiments, step (a) may be performed by dissolving the
liquid oil(s) and phospholipid(s) in ethanol, and then removing ethanol (e.g.,
via
vacuum) until the residual ethanol is less than 1% of the dry weight to obtain
a
clear oil solution.
In certain embodiments, step (c) may be performed by adding the
aqueous solution of step (b) to the mixture of step (a) to form a primary
emulsion.
The aqueous solution may further contain buffer and/or tonicity modifier(s).
The
formation of the primary emulsion may be performed or facilitated by the use
of
mechanical homogenization (e.g., high shear mixing, high pressure extrusion,
and
microfluidization) or other suitable techniques. In certain embodiments, the
pH of
the primary emulsion is adjusted to about 4. The above-described primary
emulsion may be further refined by cycling through a microfluidizer
homogenizer or
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a similar apparatus to obtain a stable emulsion having fairly uniform oil
droplet
sizes. The resulting refined emulsion may be filter sterilization, for
example,
through a 0.22-micron sterile filter.
An exemplary method of preparing an oil-in-water emulsion of
vancomycin hydrochloride is provided in Examples 3 and 6.
Besides being ready-to-use oil-in-water emulsions, the freeze-stable
vancomycin compositions of the present invention can be frozen for storage and
thawed at a later date before administration. The frozen formulations prevent
or
reduce the rapid degradation of vancomycin by hydrolysis.
In one aspect, the present application provides a frozen composition
that comprises vancomycin or a pharmaceutically acceptable salt or analog
thereof, liquid oil(s), phospholipid(s), dextrose, and water. The compositions
may
be prepared by freezing the freeze-stable oil-in-water emulsions provided
herein.
The resulting compositions may be thawed to form an oil-in-water emulsion
suitable for administration (e.g., injection).
"Frozen emulsion" is an emulsion that is stored at a sub-ambient
temperature (e.g., -20 C to -10 C) at which the aqueous phase of the emulsion
is
completely or partially crystallized (e.g., forms ice).
An exemplary frozen composition comprises: (i) vancomycin at a
concentration of about 0.5% by weight or a pharmaceutically acceptable salt or
analog of vancomycin at an equivalent concentration; (ii) one or more liquid
oils at
a total concentration of about 2% to about 4% by weight; (iii) one or more
phospholipids at a total concentration of about 1% to about 5% by weight,
wherein
the weight ratio of the total phospholipid(s) to the total liquid oil(s) is at
least 0.5:1;
(iv) about 5% to about 6% dextrose by weight; and (v) water.
Another exemplary frozen composition comprises: (1) vancomycin
hydrochloride at a concentration of about 0.5% by weight, (2) soybean oil at a
concentration of about 1%to about 2% by weight, (3) medium chain triglyceride
at
a concentration of about 1% to about 2% by weight, (4) lecithin at a
concentration
of about 2% to about 4% by weight, and (5) dextrose at a concentration of
about
5% to about 6% by weight.
A further exemplary frozen composition comprises: (1) vancomycin
hydrochloride at a concentration of about 0.5% by weight, (2) soybean oil and
medium chain triglyceride, wherein the total concentration of soybean oil and
medium chain triglyceride is about 2% by weight, the weight ratio of soybean
oil to
medium chain triglyceride is between 2:1 to 1:1, (3) lecithin at a
concentration of
about 2% by weight, and (4) dextrose at a concentration of about 5% by weight.
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In certain embodiments, when thawed, the frozen composition re-
forms an oil-in-water emulsion with an average diameter of oil droplets no
more
than about 200 nm.
In certain embodiments, when thawed, the frozen composition re-
forms an oil-in-water emulsion with a PFAT5 of less than about 0.05.
In certain embodiments, when thawed, the frozen composition re-
forms an oil-in-water emulsion with an average diameter of oil droplets no
more
than about 200 nm and a PFAT5 of less than about 0.05.
In certain embodiments, the frozen formulation is physically stable,
chemically stable, or both physically and chemically stable at room
temperature for
at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.
A frozen formulation is "physically stable" if it may be stored in the
frozen state for a defined period of time without increase in average droplet
size of
the oil-in-water emulsion thawed from the frozen formulation after storage by
more
than 50%, or evidence of phase separation or oil droplet aggregation
(coalescence) of the thawed emulsion. In certain embodiments, the average size
of oil droplets of the emulsion re-formed from a frozen formulation of the
present
invention after being stored in the frozen state for at least 1, 2, 3, 4, 5,
6, 7, 8, 9,
10, 11, or 12 months does not increase or does not increase by more than about
10%, 20%, 25%, 30%, or 40% compared with that of the freeze-stable oil-in-
water
emulsion from which the frozen formulation has been prepared.
A frozen formulation is "chemically stable" if the vancomycin
concentration in the formulation does not change by about 20% under
appropriate
storage conditions for a defined period of time. In certain embodiments, the
vancomycin concentration in an emulsion re-formed from the frozen formulation
does not change or does not change by about 5%, 10%, 15% or 20% in the frozen
state compared with that of the freeze-stable oil-in-water emulsion from which
the
frozen formulation has been prepared for at least 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, or
12 months.
In certain embodiments, the one or more liquid oils of the frozen
composition comprise a vegetable oil and a medium chain_triglyceride.
In certain embodiments, the frozen composition does not further
comprise a compound that increases the amount of vancomycin in the oil
droplets
of the emulsion formed by thawing the frozen composition.
In certain embodiments, when the frozen composition is thawed to
form an oil-in-water emulsion, no more than about 70%, 60%, 50%, 40%, 30%, or
20% of vancomycin is present in the oil droplets of the oil-in-water emulsion.
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In a related aspect, oil-in-water emulsions of vancomycin re-formed
by thawing the frozen formulations described herein are also provided.
In certain embodiments, the re-formed oil-in-water emulsions have
the same chemical and physical characteristics as described above for the
freeze-
stable oil-in-water emulsions. For example, in certain embodiments, the re-
formed
oil-in-water emulsion has an average diameter of oil droplets no more than
about
200 nm, and/or a PFAT5 of less than about 0.05. In certain embodiments, the re-
formed oil-in-water emulsion is isotonic and ready for use. In certain
embodiments, the re-formed oil-in-water emulsion does not further comprise a
compound that increases the amount of vancomycin in the oil droplets of the re-
formed emulsion. In certain embodiments, no more than about 70%, 60%, 50%,
40%, 30%, or 20% of vancomycin is present in the oil droplets in the re-formed
oil-
in-water emulsion.
The vancomycin formulations described herein may be used to treat
or prevent (i.e., reduce or eliminate the risk of) infection for which
vancomycin is
effective, including treating or preventing gram-positive infection, treating
peritoneal dialysis-associated peritonitis in patients with end-stage renal
disease,
treating pseudomembranous colitis due to Clostridium difficile or
Staphylococcus
aureus enterocolitis, surgical prophylaxis in patients allergic to penicillin
or other
beta-lactam antibiotics, bacterial endocarditis prophylaxis in patients
undergoing
gastrointestinal/genitourinary procedures and allergic to ampicillin,
amoxicillin,
penicillin, or other beta-lactam antibiotics, treatment of meningitis or
ventriculitis,
nosocomial bacteremia prophylaxis in neonates receiving total parenteral
nutrition.
The vancomycin oil-in-water formulations of the present invention
(either directly prepared or re-formed from frozen formulations) may be
administered to a subject (e.g., human or other mammals) in need thereof at a
pharmaceutically effective amount by various routes, including but not limited
to,
intravenous, intramuscular, intra-arterial, intrathecal, intraocular,
subcutaneous,
intraarticular, intra-peritoneal, oral, topical, intravaginal, and ophthalmic
administration.
"Pharmaceutically effective amount" refers to an amount of an
antibiotic (i.e., vancomycin or clarithromycin) oil-in-water emulsion that is
sufficient
for treating infection.
The vancomycin oil-in-water emulsions provided herein can be
administered in a single daily dosage or in multiple doses per day. Other
periodic
treatment protocols may also be adopted. The treatment may require
administration over extended periods of time, such as for several days or for
from
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about one to four weeks. The amount per administered dose or the total amount
administered will depend on various factors such as the route and frequency of
administration, the nature and severity of the infection, the age, sex,
weight, and
general health of the patient,. and can be determined by a physician according
to
principles of treatment well known in the antibiotic art.
In certain embodiments, the oil-in-water emulsions provided herein
may be administered intravenously to an adult human patient at a daily dose of
about 2 grams of vancomycin divided as about 500 mg every 6 hours or about 1
gram every 12 hours. Other exemplary dosages for intraveneous administration
of
the oil-in-water emulsions provided herein include about 5 mg to 10 mg of
vancomycin per kg of body weight every six hours or about 10 mg to 20 mg of
vancomycin per kg of body weight every twelve hours.
Clarithromycin Freeze-Stable or Frozen Formulations
"Clarithromycin" refers to 6-O-methyl-erythromycin (see, U.S. Pat.
No. 4,331,803) with a structure shown below
H H3C H3C~
, ~ CH3 ~'. H OCH3 ---~
HO -0 H H_ Ci-13
H '-. 0 ..-CHa _=H 0
H,3G="" CH3 N(ct-ta)z
H 0 H CH2CH3 O
OH
HO 0
CH3
OCH3
CH3
"Clarithromycin" also refers to se.misynthetic derivatives of
clarithromycin (e.g., pharmaceutically acceptable salts and esters of
clarithromycin).
In another aspect of the present invention, a freeze-stable oil-in-
water emulsion is provided that comprises: (i) clarithromycin at a
concentration of
at least about 0.5% by weight or a pharmaceutically acceptable salt or ester
of
clarithromycin at an equivalent concentration; (ii) one or more liquid oils at
a total
concentration of about 2% to about 4% by weight; (iii) one or more
phospholipids
at a total concentration of about 1% to about 5% by weight, wherein the weight
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ratio of the total phospholipid(s) to the total liquid oil(s) is at least
0.5:1; (iv) about
5% to about 6% dextrose by weight; and (v) water.
"Pharmaceutically acceptable salts and esters" refers to salts and
esters which are, within the scope of sound medical judgment, suitable for use
in
contact with the tissues of humans and lower animals without undue toxicity,
irritation, allergic response, and the like, and effective for their intended
use in the
chemotherapy and prophylaxis of antimicrobial infections. Among the more
common pharmaceutically acceptable salts and esters of clarithromycin are
acetate, estolate (lauryl sulfate salt of the propionate ester), ethyl
succinate,
gluceptate (glucoheptonate), lactobionate, stearate, and hycrochloride forms.
Other acid salts used in the pharmaceutical art are the following: adipate,
alginate,
aspartate, benzoate, benzene-sulfonate, bisulfate, butyrate, citrate,
camphorate,
camphorsulfonate, cyclopentaneproiponate, digluconate, dodecylsulfate,
ethanesulfonate, fumarate, gluconate, glycerophosphate, hemisulfate,
heptaonate,
hexanoate, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactate,
maleate, methanesulfonate, 2-naphthalene-sulfonate, nicotinate, oxalate,
pamoate, pantothenate, pectinate, persutfate, 3-pheylpropionate, picrate,
pivalate,
propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate. Basic
nitrogen-containing groups can be quaternized with such agents as lower alkyl
halides, such as methyl, ethyl, propyl and butyl chloride, bromides and
iodides;
dialkyl sulfates like dimethyl, diethyl, dibutly, and diamyl sulfates; long
chain
halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and
iodides;
aralkyl halides like benzyl and phenethyl bromides and others. Water or oil-
soluble
or dispersible products are thereby obtained.
A pharmaceutically acceptable salt of clarithromycin is at a
concentration "equivalent" to a specified concentration of clarithromycin if
at that
concentration, the clarithromycin salt contains the same amount of
clarithromycin
free base as clarithromycin at the specified concentration.
A pharmaceutically acceptable ester of clarithromycin is at a
concentration "equivalent" to a specified concentration of clarithromycin if
at that
concentration, the clarithromycin ester is as effective as clarithromycin at
the
specified concentration in treating or preventing infection.
In the freeze-stable oil-in-water emulsions provided herein,
clarithromycin is present at a concentration of at least about 0.5%. In
certain
embodiments, clarithromycin is present at a concentration of at least about
1%,
2%, or 3%.
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The freeze-stable oil-in-water clarithromycin emulsions provided
herein contain one or more liquid oils at a total concentration of about 2% to
about
4% by weight, including any values from 1.8% to 4.4%.
In certain embodiments, the oil component of the oil-in-water
clarithromycin emulsions provided herein comprises a monoglyceride, a
diglyceride, a triglyceride, or a mixture thereof. In certain embodiments, the
oil
component comprises an ester formed between one or more fatty acids and an
alcohol other than glycerol.
In certain embodiments, the one or more liquid oils of the oil-in-water
clarithromycin emulsions comprise a vegetable oil, such as soybean oil.
In certain embodiments, combinations of vegetable oil and MCT oil
are used in the present invention. Generally, MCT oil is limited to at most
50% by
weight in the combinations of vegetable oil and MCT.
In certain embodiments, the one or more phospholipids of the freeze-
stable oil-in-water clarithromycin emulsions comprise lecithin, such as soy
lecithin
or egg lecithin.
The freeze-stable oil-in-water clarithromycin emulsions provided
herein comprise one or more phospholipids at a total concentration of about 2%
to
about 5% by weight (including any value between 1.8% and 5.5%).
The weight ratio of the total phospholipid(s) to the total liquid oil(s) in
the freeze-stable oil-in-water clarithromycin emulsions provided herein is at
least
0.5:1. In certain embodiments, the weight ratio of the total phospholipid(s)
to the
total liquid oil(s) is 1:1.
The freeze-stable oil-in-water clarithromycin emulsions provided
herein comprise about 5% to about 6% dextrose by weight.
In most embodiments, the freeze-stable oil-in-water clarithromycin
emulsion provided herein is isotonic and thus ready for use.
In certain embodiments, the oil droplets of the freeze-stable oil-in-
water clarithromycin emulsions are of sub-micron size. In certain embodiments,
the oil droplets of the freeze-stable oil-in-water clarithromycin emulsions
have an
average diameter of less than about 500, 450, 400, 350, 300, 250, 200, 175,
150,
125, 100, or 75 nm.
In certain embodiments, the freeze-stable oil-in-water clarithromycin
emulsions also have a PFAT5 value less than about 0.05.
In certain embodiments, the freeze-stable oil-in-water clarithromycin
emulsion has an average size of the oil droplets of no more than about 200 nm
and a PFAT5 less than about 0.05.
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In certain embodiments, the freeze-stable oil-in-water clarithromycin
emulsions provided herein do not increase their average diameters of the oil
droplets by more than about 50% after 1, 2, 3, 4, or 5 freeze-thaw cycles.
In certain embodiments, the freeze-stable oil-in-water clarithromycin
emulsions provided herein do not increase their average diameters of the oil
droplets by more than about 40%, 30%, 30%, 20%, or 10% after 1, 2, 3, 4, or 5
freeze-thaw cycles.
In certain embodiments, the freeze-stable oil-in-water clarithromycin
emulsion retains its average oil droplet size of no more than about 200 nm and
a
PFAT5 of less than about 0.05 after 1, 2, 3, 4, or 5 freeze-thaw cycles.
The freeze-stable oil-in-water clarithromycin emulsions disclosed
herein are stable when stored at a frozen state (e.g., at -20 C) for an
extended
period of time (e.g., for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months) as
described
in more detail below.
In certain embodiments, the pH of the freeze-stable oil-in-water
emulsions of clarithromycin is about 3 to about 8 (e.g., about 6 to about 8).
In
certain embodiments, the pH is about 4 or about 7.
In certain embodiments, the oil-in-water emulsion does not further
comprise a compound that increases the amount of clarithromycin in the oil
droplets of a clarithromycin oil-in-water emulsion. Examples of such compounds
include fatty acids, N-methyl pyrrolidone, and benzyl alcohoi.
In certain embodiments, no more than about 70%, 60%, 50%, 40%,
30%, or 20% of vancomycin is present in the oil droplets of the freeze-stable
oil-in-
water clarithromycin emulsion.
In certain embodiments, some or a!l of the components other than
clarithromycin in the freeze-stable oil-in-water emulsion (e.g., an oil
component
and a phospholipid) is safe, well tolerated, and acceptable by the FDA for
intravenous injection.
In certain embodiments, some or all of the components other than
the clarithromycin in the freeze-stable oil-in-water emulsion (e.g., an oil
component
and a phospholipid) is generally regarded as safe for use in intravenous
injections
by a drug regulatory authority.
In certain embodiments, the freeze-stable oil-in-water emulsions are
vein non-irritable, injectable, and/or biocompatible.
An exemplary freeze-stable oil-in-water clarithromycin emulsion
comprises: (1) clarithromycin at a concentration of about 1% to about 5% by
weight, (2) soybean oil and medium chain triglyceride, wherein the total
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concentration of soybean oil and medium chain triglyceride is about 2% by
weight,
and the weight ratio of soybean oil to medium chain triglyceride is between
2:1 to
1:1, (3) lecithin at a concentration of about 2% by weight, and (4) dextrose
at a
concentration of about 5% to about 6% by weight.
Another exemplary freeze-stable oil-in-water clarithromycin emulsion
comprises: (1) clarithromycin at a concentration of about 5% by weight, (2)
soybean oil at a concentration of about 1% by weight, (3) a medium chain
triglyceride at a concentration of about 1 % by weight, (4) lecithin at a
concentration
of about 2% by weight, and (5) dextrose at a concentration of about 5% by
weight.
The present invention also provides methods for preparing freeze-
stable oil-in-water emulsions of clarithromycin described herein. Such
emulsion
compositions may be prepared by (a) forming a mixture that comprises (i) one
or
more liquid oils (e.g., a vegetable oil, or a combination of a vegetable oil
and a
medium chain triglyceride) and (ii) one or more phospholipids, (b) forming a
mixture that comprises (i) a pharmaceutically effective amount of
clarithromycin or
a pharmaceutically acceptable salt or ester thereof, (ii) dextrose, and (iii)
water,
and (c) forming an oil-in-water emulsion with the mixtures of step (a) and
(b).
In certain embodiments, step (a) may be performed by dissolving the
liquid oil(s) and phospholipid(s) in ethanol, and then removing ethanol (e.g.,
via
vacuum) until the residual ethanol is less than 1% of the dry weight to obtain
a
clear oil solution.
In certain embodiments, step (c) may be performed by adding the
aqueous solution of step (b) to the mixture of step (a) to form a primary
emulsion.
The aqueous solution may, further contain buffer and/or tonicity modifier(s).
The
formation of the primary emulsion may be performed or facilitated by the use
of
mechanical homogenization (e.g., high shear mixing, high pressure extrusion,
and
microfluidization) or other suitable techniques. In certain embodiments, the
pH of
the primary emulsion is adjusted to about 5 to about 9 (e.g., about 7). The
above-
described primary emulsion may be further refined by cycling through a
microfluidizer homogenizer or a similar apparatus to obtain a stable emulsion
having fairly uniform oil droplet sizes. The resulting refined emulsion may be
filter
sterilization, for example, through a 0.22-micron sterile filter.
An exemplary method of preparing an oil-in-water emulsion of
clarithromycin is provided in Example 9.
Besides being ready-to-use oil-in-water emulsions, the freeze-stable
clarithromycin compositions of the present invention can be frozen for storage
and
thawed at a later date before injection.
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In one aspect, the present application provides a frozen composition
that comprises clarithromycin or a pharmaceutically acceptable salt or ester
thereof, liquid oil(s), phospholipid(s), dextrose, and water. The compositions
may
be prepared by freezing the freeze-stable oil-in-water emulsions provided
herein.
The resulting compositions may be thawed to form an oil-in-water emulsion
suitable for injection.
An exemplary frozen composition comprises: (i) clarithromycin at a
concentration of at least about 0.5% by weight or a pharmaceutically
acceptable
salt or ester of clarithromycin at an equivalent concentration; (ii) one or
more liquid
oils at a total concentration of about 2% to about 4% by weight; (iii) one or
more
phospholipids at a total concentration of about 1 % to about 5% by weight,
wherein
the weight ratio of the total phospholipid(s) to the total liquid oil(s) is at
least 0.5:1;
(iv) about 5% to about 6% dextrose by weight; and (v) water.
Another exemplary frozen composition comprises: clarithromycin at a
concentration of about 1% to about 5% by weight, (2) soybean oil and medium
chain triglyceride, wherein the total concentration of soybean oil and medium
chain
triglyceride is about 2% by weight, and the weight ratio of soybean oil to
medium
chain triglyceride is between 2:1 to 1:1, (3) lecithin at a concentration of
about 2%
by weight, and (4) dextrose at a concentration of about 5% to about 6% by
weight.
A further exemplary frozen composition comprises: (1) clarithromycin
at a concentration of about 5% by weight, (2) soybean oil at a concentration
of
about 1% by weight, (3) a medium chain triglyceride at a concentration of
about
1% by weight, (4) lecithin at a concentration of about 2% by weight, and (5)
dextrose at a concentration of about 5% by weight.
In certain embodiments, when thawed, the frozen composition re-
forms an oil-in-water emulsion with an average diameter of oil droplets no
more
than about 200 nm.
In certain embodiments, when thawed, the frozen composition re-
forms an oil-in-water emulsion with a PFAT5 less than about 0.05.
In certain embodiments, when thawed, the frozen composition re-
forms an oil-in-water emulsion with an average diameter of oil droplets no
more
than about 200 nm and a PFAT5 less than about 0.05.
In certain embodiments, the frozen formulation is physically stable,
chemically stable, or both physically and chemically stable at room
temperature for
at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.
In certain embodiments, the average size of oil droplets of the
emulsion re-formed from a frozen formulation disclosed herein after being
stored in
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the frozen state for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months
does not
increase or does not increase by more than about 10%, 20%, 25%, 30%, or 40%
compared with that of the freeze-stable oil-in-water emulsion from which the
frozen
formulation has been prepared.
In certain embodiments, the clarithromycin concentration in an the
emulsion re-formed from a frozen formulation disclosed herein after being
stored in
the frozen state for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months
does not
change or does not change by about 5%, 10%, 15% or 20% in the frozen state
compared with that of the freeze-stable oil-in-water emulsion from which the
frozen
formulation has been prepared.
In certain embodiments, the one or more liquid oils of the frozen
composition comprise a vegetable oil and a medium chain triglyceride.
In certain embodiments, the frozen composition does not further
comprise a compound that increases the amount of vancomycin in the oil
droplets
of the emulsion.
In certain embodiments, when the frozen composition is thawed to
form an oil-in-water emulsion, no more than about 70%, 60%, 50%, 40%, 30%, or
20% of clarithromycin is present in the oil droplets of the oil-in-water
emulsion.
In a related aspect, oil-in-water emulsions of clarithromycin re-formed
by thawing the frozen formulations described herein are also provided.
In certain embodiments, the re-formed oil-in-water emulsions have
the same chemical and physical characteristics as described above for the
freeze-
stable oil-in-water emulsions. For example, in certain embodiments, the re-
formed
oil-in-water emulsion has an average diameter of oil droplets no more than
about
200 nm, and/or a PFAT5 less than about 0.05. In certain embodiments, the re-
formed oil-in-water emulsion is isotonic and ready for use. In certain
embodiments, the re-formed oil-in-water emulsion does not further comprise a
compound that increases the amount of clarithromycin in the oil droplets of
the re-
formed emulsion. In certain embodiments, no more than about 70%, 60%, 50%,
40%, 30%, or 20% of clarithromycin is present in the oil droplets in the re-
formed
oil-in-water emulsion.
The clarithromycin formulations provided herein may be used to treat
or reduce the risk of infection for which clarithromycin is effective,
including
infections caused by bacterial, such as pneumonia, bronchitis, and ear, lung,
sinus, stomach, skin and throat infections.
The clarithromycin oil-in-water formulations of the present invention
(either directly prepared or re-formed from frozen formulations) may be
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administered to a subject (e.g., human or other mammals) in need thereof at a
pharmaceutically effective amount by various routes, including but not limited
to,
intravenous, intramuscular, intra-arterial, intrathecal, intra-ocular,
subcutaneous,
intra-articular, oral, topical, intravaginal and intra-peritoneal
administration.
The clarithromycin oil-in-water emulsions provided herein can be
administered in a single daily dosage or in multiple doses per day. Other
periodic
treatment schemes may also be used. The treatment may require administration
over extended periods of time, such as for several days or for from about one
to
four weeks. The amount per administered dose or the total amount administered
will depend on various factors such as the route and frequency of
administration,
the nature and severity of the infection, the age, sex, weight, and general
health of
the patient, and can be determined by a physician according to principles of
treatment well known in the antibiotic art. For example, in certain
embodiments,
the oil-in-water emulsions provided herein may be administered intravenously
to a
human adult patient at a dose of about 250 mg to about 750 mg every 12 hours.
Lyophilizable Oil-in-Water Emulsions and Lyophilized Formulations
In another aspect of the present invention, lyophilizable oil-in-water
emulsions are provided for vancomycin and clarithromycin. Such emulsions may
have one or more of the following advantages: (1) they may be lyophilized for
storage and remain stable during storage, (2) they contain sub-micron oil
droplets
and may be filter sterilized, (3) the lyophilized emulsions may be
reconstituted to
oil-in-water emulsions with sub-micron oil droplets, and the reconstituted
emulsions may also be filter sterilized, (4) the oil-in-water emulsions that
are either
directly prepared or reconstituted from the lyophilized emulsions are non-vein
irritating, and (5) the oil-in-water emulsions do not contain any additional
compounds (e.g., stabilizers), which avoids any side effects of such
compounds_
The word "reformed" is used interchangeably with "reconstituted" in
connection with describing oil-in-water emulsions formed from lyophilized
compositions.
Vancomycin Lyophilizable and Lyophilized Formulations
In one aspect, the present invention provides a lyophilizable oil-in-
water emulsion that comprises: (i) at least 15 mglml vancomycin or a
pharmaceutically acceptable salt or analog thereof, (ii) one or more liquid
oils at a
total concentration of about 2% to about 10% by weight, (iii) one or more
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phospholipids at a total concentration of about 1% to about 10% by weight, and
(iv)
dextrose at a concentration of at least about 10% by weight.
In certain embodiments, vancomycin or a pharmaceutically
acceptable salt or analog is present in the oil-in-water emulsion at a
concentration
at least about 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50
mg/mi.
The amount of the total oil component in the lyophilizable
vancomycin emulsions provided herein may be within a range of about 1% to
about 20% (e.g., about 2% to about 10%) by weight. In certain embodiments, the
total concentration of the oil component is about 2%, 3%, 4%, 5%, 6%, 7%, 8%,
9 Ao, 10%, 12%, 15%, 18%, or 20% by weight.
In certain embodiments, the weight ratio of vegetable oil to MCT oil in
the oil-in-water vancomycin emulsion is within a range of about 9:1 to about
1:1 by
weight. In certain embodiments, the weight ratio of the vegetable oil to MCT
oil is
abut 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 or 1:1.
The amount of phospholipids, by weight, in the vancomycin
emulsions provided herein may be within a range of about 1% to about 10%. In
certain embodiments, the phospholipids in the emulsions are at a
concentration, by
weight, about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.
The lyophilizable oil-in-water emulsions of vancomycin also comprise
dextrose at a concentration of at least about 10% by weight. In certain
embodiments, the concentration of dextrose in the oil-in-water emulsions is
about
12%, 14%, 15%, 18 fo, 20%, 22%, 24%, 25%, 28%, or 30% by weight.
The presence of dextrose in the oil-in-water emulsions prevents the
droplets from forming aggregates or phase separation during lyophilization and
reconstitution, and allows the resulting lyophilized formulations to be
reconstituted
with water quickly (e.g., within about 5 minutes or less) to re-form oil-in-
water
emulsions with average diameter of droplets no greater than about 1 micron. In
addition, because vancomycin is a high dose drug, a relatively large amount of
cryoprotectants is required in the oil-in-water emulsions to ensure that the
formulations after being lyophilized may be reconstituted to re-form oil-in-
water
emulsions with micron size or sub-micron size droplets. Using dextrose as the
cryoprotectant avoids the safety or biological activity concerns associated
with
other cryoprotectants such as mannitol when administered intravenously in a
relatively large quantity.
In certain embodiments, the average size of oil droplets of a
lyophilizable emulsion provided herein does not increase by more than about
10%,
20%, 25%, 30%, 40%, 50%, 75%, 100%, 125%, 150%, 175%, or 200% under
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appropriate storage conditions (e.g., at 2-8 C) for at least 1, 2, 3, 4, 5, 6,
9, 12, 15,
18, 20, 25, or 30 days.
In certain embodiments, the vancomycin concentration in a
iyophilizabte emulsion of the present invention does not change by about 5%,
10%, 15% or 20% under appropriate storage conditions (e.g., at 2-8 C) for at
least
1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days.
In certain embodiments, the oil droplets of the lyophilizable
compositions provided herein have an average diameter of less than about 500,
450, 400, 350, 300, 250, 200, 175, 150, 125, 100, 75, or 50 nm.
In certain embodiments, the oil-in-water emulsion has a PFAT5 of
less than about 0.05. In certain embodiments, the oil-in-water emulsion has an
average
diameter of oil droplets of less than about 200 nm and a PFAT5 of less than
about
0.05.
In certain embodiments, the oil-in-water emulsion is sterilized. For
instance, the oil-in-water emulsion with an average oil droplet diameter of no
more
than about 200 nm may be sterilized via a 0.2 m filter.
In certain embodiments, the pH of the oil-in-water emulsions of
vancomycin is about 3 to about 8). in certain embodiments, the pH is about 7
or
about 7.5.
In certain embodiments, the lyophilizable oil-in-water emulsion does
not further comprise a compound that increases the amount of vancomycin in the
oil droplets of the emulsion.
In certain other embodiments, although the lyophilizable oil-in-water
emulsion may further comprise a compound that increases the amount of
vancomycin in the oil droplets, the stability (physical and/or chemical
stability) of
the emulsion does not require the presence of such a compound.
In certain embodiments, less than about 70%, 60%, 50%, 40%, 30%,
or 20% of vancomycin is in the oil droplets of the oil-in-water emulsions.
"Lyophilizable oil-in-water emulsion" refers to an oil-in-water emulsion
("original oil-in-water emulsion") that can be lyophilized to form a lyophile,
and the
average oil droplet size of the oil-in-water emulsion reconstituted from the
lyophile
does not increase by 50% compared to that of the original oil-in-water
emulsion.
In certain embodiments, the lyophilizable oil-in-water emulsion
retains its average oil droplet size of no more than about 200 nm and a PFAT5
of
less than about 0.05 after being lyophilized to form a lyophile and then
reconstituted from the lyophile.
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Another exemplary lyophilizable oil-in-water emulsion comprises:
vancomycin hydrochloride at a concentration of about 1 % to about 3% by
weight,
medium chain triglyceride at a concentration of about 1% to about 5% by
weight,
vegetable oil at a concentration of about 1% to about 5% by weight, lecithin
at a
concentration of about 1% to 4% by weight, and dextrose of about 15% to 25% by
weight.
The present invention also provides methods for preparing
lyophilizable oil-in-water emulsions of vancomycin described herein. Such
emulsion compositions may be prepared by (a) forming a mixture that comprises
(i) one or more liquid oils (e.g., a vegetable oil, or a combination of a
vegetable oil
and a medium chain triglyceride) and (ii) one or more phospholipids, (b)
forming a
mixture that comprises: (i) a pharmaceutically effective amount of vancomycin
or a
pharmaceutically acceptable salt or analog thereof, (ii) dextrose, and (iii)
water,
and (c) forming an oil-in-water emulsion with the mixtures of step (a) and
(b).
In certain embodiments, step (a) may be performed by dissolving the
liquid oil(s) and phospholipid(s) in ethanol, and then removing ethanol (e.g.,
via
vacuum) until the residual ethanol is less than 1% of the dry weight to obtain
a
clear oil solution.
In certain embodiments, step (c) may be performed by adding the
aqueous solution of step (b) to the mixture of step (a) to form a primary
emulsion.
The aqueous solution may further contain buffer, stabilizer(s) and/or tonicity
modifier(s). The formation of the primary emulsion may be performed or
facilitated
by the use of mechanical homogenization (e.g., high shear mixing, high
pressure
extrusion, and microfluidization) or other suitable techniques. In certain
embodiments, the pH of the primary emulsion is adjusted to about 6 to about 8
(e.g., about 7.4). The above-described primary emulsion may be further refined
by
cycling through a microfluidizer homogenizer or a similar apparatus to obtain
a
stable emulsion having fairly uniform oil droplet sizes. The resulting refined
emulsion may be filter sterilization, for example, through a 0.22-micron
sterile filter.
An exemplary method of preparing an oil-in-water emulsion of
vancomycin hydrochloride is provided in Example 1.
Besides being ready-to-use oil-in-water emulsions, the lyophilizable
vancomycin compositions provided herein can also be lyophilized and then
reconstituted at a later date, prior to injection, by dilution with water, to
reform the
oil-in-water emulsion. As a lyophile, the formulations of vancomycin in
accordance
with the invention prevent the rapid degradation of vancomycin by hydrolysis,
which occurs in the presence of water.
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In another aspect, the present application provides a lyophilized
composition that comprises vancomycin or a pharmaceutically acceptable salt or
analog thereof, liquid oil(s), phospholipid(s), and dextrose. The compositions
may
be prepared by removal of water from the above-described oil-in-water
emulsions.
The resulting compositions may be rehydrated with water or a buffer solution
to
form an oil-in-water emulsion suitable for injection. The average diameter of
the
re-formed emulsion droplets is no greater than about 1 micron.
In certain embodiments, the average diameter of the re-formed
emulsion droplets is no greater than about 1000, 900, 800, 700, 600, 500, 450,
400, 350, 300, 250, 200, 175, 150, 125, 100, 75, or 50 nm.
)n, certain embodiments, the average size of oil droplets of the
emulsion reconstituted from a lyophilized formulation of the present invention
after
the lyophilized formulation is stored under appropriate storage conditions
(e.g., at -
C) for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, or
24
15 months does not increase by more than about 10%, 20%, 25%, 30%, 40%, 50%,
75%, 100%, 125%, 150%, 175%, or 200% compared with that of the lyophilizable
oil-in-water emulsion from which the lyophilized formulation has been
prepared.
In certain embodiments, the lyophilized formulation does not further
comprise a compound that increases the amount of vancomycin in the oil
droplets
20 of the emulsion reconstituted from the lyophilized formulation.
In certain embodiments, when the lyophilized formulation is
reconstituted to form an oil-in-water emulsion, no more than about 70%, 60%,
50%, 40%, 30%, or 20% of vancomycin is present in the oil droplets of the oil-
in-
water emulsion.
In certain embodiments, the vancomycin concentration in the
emulsion reconstituted from a lyophilized formulation provided herein does not
change by about 5%, 10%, 15% or 20% afterthe lyophilized formulation has been
stored under appropriate storage conditions (e.g., at-20 C) for at least 1, 2,
3, 4,
5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, or 24 months compared with that
of the
lyophilizable oil-in-water emulsion from which the lyophilized formulation has
been
prepared.
In certain embodiments, the present application provides a
lyophilized composition comprising: (i) vancomycin or a pharmaceutically
acceptable salt or analog thereof at a concentration of about 5% to 10% by
weight,
(ii) one or more liquid oils at a total concentration of about 10% to 20% by
weight,
(iii) one or more phospholipids at a total concentration of about 10% to 20%
by
weight, and (iv) dextrose at a concentration 50% to 80% by weight.
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The lyophilized vancomycin formulations of the present invention
may be prepared by lyophilizing the oil-in-water emulsions as described herein
via
any appropriate lyophilization technique (e.g., via a shelf freeze-dryer).
"Lyophilization" or "freeze drying" is a process in which water is
removed from a product after it is frozen and placed under a vacuum, allowing
the
ice to change directly from solid to vapor without passing through a liquid
phase.
The process consists of three separate, unique, and interdependent processes:
freezing, primary drying (sublimation), and secondary drying (desorption).
The lyophilized formulation may be reconstituted with an appropriate
amount of an aqueous solution (e.g., water, a buffer solution, or an aqueous
solution with other additives such as tonicity modifiers) to re-form an oil-in-
water
emulsion that comprises vancomycin or a pharmaceutically acceptable salt or
analog thereof at a pharmaceutically effective concentration.
"Pharmaceutically
effective concentration" refers to a concentration of vancomycin or its
pharmaceutically acceptable salt or analog in an oil-in-water emulsion that is
sufficient in treating infections or other diseases that vancomycin or its
pharmaceutically acceptable salt or analog is effective when the oil-in-water
emulsion is administered.
In certain embodiments, the average oil droplet size of the
reconstituted oil-in-water emulsion does not increase by more than about 10%,
20%, 30%, 40 fo, 50%, 60%, 70%, 70%, 80%, 90%, or 100% for at least about 1,
2,
3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 days under appropriate storage condition
(e.g., at 2-
8 C).
In certain embodiments, the concentration of vancomycin in the
reconstituted oil-in-water emulsion does not change by 5%,10%, 15%, or 20% for
at least about 1, 2, 3, 4, 5, fi, 7, 8, 9, 10, 11, or 12 days under
appropriate storage
condition (e.g., at 2-8 C).
In certain embodiments, the reconstituted oil-in-water emulsion is
non-vein irritating.
In certain embodiments, the reconstituted oil-in-water emulsion has a
pH of about 6 to about 8 (e.g., about 7 or about 7.5).
In certain embodiments, the reconstituted oil-in-water emulsions
have the same chemical and physical characteristics as described above for the
lyophilizable oil-in-water emulsions. For example, in certain embodiments, the
reconstituted oil-in-water emulsion has an average diameter of oil droplets no
more
than about 200 nm, and/or a PFAT5 of less than about 0.05. In certain
embodiments, the reconstituted oil-in-water emulsion is isotonic and ready for
use.
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In certain embodiments, the reconstituted oil-in-water emulsion does not
further
comprise a compound that increases the amount of vancomycin in the oil
droplets
of the reconstituted emulsion. In certain embodiments, no more than about 70%,
60%, 50%, 40%, 30%, or 20% of vancomycin is present in the oil droplets in the
reconstituted oil-in-water emulsion.
The lyophilizable vancomycin formulations as well as oil-in-water
emulsions reconstituted from lyophilized formulations provided herein may be
used
to treat or prevent infection for which vancomycin is effective. More
specifically,
they may be administered to a subject (e.g., human or other mammals) in need
thereof at a pharmaceutically effective amount by various routes, including
but not
limited to, intravenous, intramuscular, intra-arterial, intrathecal,
intraocular,
subcutaneous, intra-articular, intra-peritoneal, oral, topical, intravaginal
and
ophthalmic administration.
Clarithromycin Lyophilizable and Lyophilized Formulations
In another aspect, a lyophilizable oil-in-water emulsion is provided
that comprises: (i) clarithromycin at a concentration of at least about 1% by
weight
or a pharmaceutically acceptable salt or ester thereof at an equivalent
concentration, (ii) one or more liquid oils at a total concentration of about
2% to
about 10% by weight, (iii) one or more phospholipids at a total concentration
of
about 1% to about 10% by weight, and (iv) dextrose at a concentration of at
least
about 10% by weight.
In certain embodiments, clarithromycin or a pharmaceutically
acceptable salt or ester thereof is present in the oil-in-water emulsion at a
concentration at least about 0.5% by weight. In certain embodiment, the
concentration of clarithromycin or a pharmaceutically acceptable salt or ester
thereof in the emulsion is about 1%, 1.5%, 2%, 2.5%, 3%, 4%, or 5%.
The content of the total oil component in the lyophilized
clarithromycin emulsions provided herein may be within a range of about 1% to
about 20% (e.g., about 2% to about 10%) by weight. In certain embodiments, the
total concentration of the oil component is about 2%, 3%, 4%, 5%, 6%, 7%, 8%,
9%, 10%, 12%, 15%, 18%, or 20% by weight.
In certain embodiments, the weight ratio of vegetable oil to MCT oil in
the oil-in-water emulsion is within a range of about 91 to about 1:1, by
weight. ln
certain embodiments, the weight ratio of the vegetable oil to MCT oil is abut
9:1,
8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1 or 1:1.
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The amount of phospholipids, by weight, in the lyophilized
clarithromycin emulsions provided herein may be within a range of about 1% to
about 10%. In certain embodiments, the phospholipids in the emulsions are at a
concentration, by weight, about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%.
The lyophilized oil-in-water emulsions of clarithromycin also comprise
dextrose at a concentration of at least about 10% by weight. In certain
embodiments, the concentration of dextrose in the oil-in-water emulsions is
about
12%, 14%, 15%, 18%, 20%, 22%, 24%, 25%, 28%, or 30% by weight.
In certain embodiments, some or all of the components other than
clarithromycin or its pharmaceutically acceptable salt or ester in the oil-in-
water
emulsion (e.g., an oil component, a phospholipid, a stabilizer, and a tonicity
modifier) is safe, well tolerated, and acceptable by the FDA for intravenous
injection.
In certain embodiments, some or all of the components other than
clarithromycin or its pharmaceutically acceptable salt or ester in the oil-in-
water
emulsion (e.g., an oil component, an emulsifier, a stabilizer, and a tonicity
modifier).
is generally regarded as safe for use in intravenous injections by a drug
regulatory
authority.
In certain embodiments, the lyophilizable oil-in-water clarithromycin
emulsions provided herein are vein non-irritable, injectable, and/or
biocompatible.
In certain embodiments, the lyophilizable oil-in-water clarithromycin
emulsions provided herein are stable physically, chemically, or both
chemically
and physically.
In certain embodiments, the average size of oil droplets of a
Iyophilizable oil-in-water clarithromycin emulsions provided herein does not
increase by more than about 10%, 20%, 25%, 30%, 40%, 50%, 75%, 100%,
125%, 150%, 175 fo, or 200% under appropriate storage conditions (e.g., at-20
C
or 2-8 C) for at least 1, 2, 3, 4, 5, 6, 9, 12, 15, 18, 20, 25, or 30 days.
In certain embodiments, the clarithromycin concentration in a
lyophilizable oil-in-water clarithromycin emulsions provided herein does not
change
by about 5%, 10%, 15% or 20% under appropriate storage conditions (e.g., at -
20 C or 2-8 C) for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days.
In certain embodiments, the oil droplets of the lyophilizable oil-in-
water clarithromycin emulsions provided herein have an average diameter of
less
than about 500, 450, 400, 350, 300, 250, 200, 175, 150, 125, 100, 75, or 50
nm.
In certain embodiments, the lyophilizable oil-in-water clarithromycin
emulsion is sterilized. For instance, the oil-in-water emulsion with an
average oil
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droplet diameter of no more than about 200 nm may be sterilized via a 0.2 m
filter.
In certain embodiments, the pH of the lyophilizable oil-in-water
clarithromycin emulsions is about 3 to about 8. In certain embodiments, the pH
is
about 7 or about 7.5.
In certain embodiments, the lyophilizable oil-in-water emulsion does
not further comprise a compound that increases the amount of clarithromycin in
the oil droplets of the emulsion.
In certain embodiments, no more than about 70%, 60%, 50%, 40 fa,
30%, or 20% of clarithromycin is present in the oil droplets of the
lyophilizable oil-
in-water emulsion.
In certain embodiments, the lyophilizable oil-in-water emulsion
retains its average oil droplet size of no more than about 200 nm and a PFAT5
of
less than about 0.05 after being lyophilized to form a lyophilized formulation
and
then reconstituted from the lyophilized formulation.
In certain embodiments, the lyophilizable oil-in-water emulsion
comprises clarithromycin at a concentration of about 1 fo to about 3% by
weight,
medium chain triglyceride at a concentration of about 1% to about 5% by
weight,
vegetable oil at a concentration of about 1% to about 5% by weight, lecithin
at a
concentration of about 1% to 10% by weight, and dextrose of about 15% to 25%
by weight.
The present invention also provides methods for preparing
lyophilizable oil-in-water emulsions of clarithromycin described herein. Such
emulsion compositions may be prepared by (a) forming a mixture that comprises
(i) one or more liquid oils (e.g., a vegetable oil, or a combination of a
vegetable oil
and a medium chain triglyceride) and (ii) one or more phospholipids, (b)
forming a
mixture that comprises (i) a pharmaceutically effective amount of
clarithromycin or
a pharmaceutically acceptable salt or ester thereof, (ii) dextrose, and (iii)
water,
and (c) forming an oil-in-water emulsion with the mixtures of step (a) and
(b).
In certain embodiments, step (a) may be performed by dissolving the
liquid oil(s) and phospholipid(s) in ethanol, and then removing ethanol (e.g.,
via
vacuum) until the residual ethanol is less than 1% of the dry weight to obtain
a
clear oil solution.
In certain embodiments, step (c) may be performed by adding the
aqueous solution of step (b) to the mixture of step (a) to form a primary
emulsion.
The aqueous solution may further contain buffer, stabilizer(s) and/or tonicity
modifier(s). The formation of the primary emulsion may be performed or
facilitated
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by the use of mechanical homogenization (e.g., high shear mixing, high
pressure
extrusion, and microfluidization) or other suitable techniques. In certain
embodiments, the pH of the primary emulsion is adjusted to about 6 to about 8
(e.g., about 7.4). The above-described primary emulsion may be further refined
by
cycling through a microfluidizer homogenizer or a similar apparatus to obtain
a
stable emulsion having fairly uniform oil droplet sizes. The resulting refined
emulsion may be filter sterilization, for example, through a 0.22-micron
sterile filter.
An exemplary method of preparing a clarithromycin oil-in-water
emulsion is provided in Example 2.
The lyophilizable clarithromycin compositions of the present invention
can also be prepared as a lyophilized formulation that can be reconstituted at
a
later date and diluted with water to reform the oil-in-water emulsion before
injection.
In one aspect, the present application provides a lyophilized
composition that comprises clarithromycin or a pharmaceutically acceptable
salt or
ester thereof, liquid oil(s), phospholipid(s), and dextrose. The compositions
may
be prepared by removal of water from the oil-in-water emulsions as described
above. The resulting compositions may be rehydrated with water or a buffer
solution to form an oil-in-water emulsion suitable for injection. The average
diameter of the re-formed emulsion droplets is no greater than about 1 micron.
In certain embodiments, the average diameter of the re-formed
emulsion droplets is no greater than about 1000, 900, 800, 700, 600, 500, 450,
400, 350, 300, 250, 200, 175, 150, 125, 100, 75, or 50 nm.
In certain embodiment, the lyophilized formulation is physically
stable, chemically stable, or both physically and chemically stable at room
temperature for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18,
20, 22, or 24
months.
In certain embodiments, the average size of oil droplets of the
clarithromycin emulsion reconstituted from a lyophilized formulation provided
herein after the lyophilized formulation has been stored under appropriate
storage
conditions (e.g., at-20 C) for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,
14, 16, 18,
20, 22, or 24 months does not increase by more than about 10%, 20%, 25%, 30%,
40%, 50%, 75%, 100%, 125%, 150%, 175%, or 200% compared with that of the
lyophilizable oil-in-water emulsion from which the lyophilized formulation has
been
prepared.
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In certain embodiments, the lyophilized formulation does not further
comprise a compound that increases the amount of clarithromycin in the oil
droplets of the reconstituted emulsion.
In certain embodiments, when the lyophilized formulation is thawed
to form an oil-in-water emulsion, no more than about 70%, 60%, 50%, 40%, 30%,
or 20% of clarithromycin is present in the oil droplets of the oil-in-water
emulsion.
In certain embodiments, the clarithromycin concentration in an oil-in-
water emulsion reconstituted from a lyophilized formulation provided herein
after
the lyophilized formulation has been stored under appropriate storage
conditions
(e.g., at -20 C) for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16,
18, 20, 22, or
24 months does not change by about 5%, 10%, 15% or 20% compared with that of
the lyophilizable oil-in-water emulsion from which the lyophilized formulation
has
been prepared.
In certain embodiments, the present invention provides a lyophilized
composition comprising: (i) clarithromycin or a pharmaceutically acceptable
salt or
ester thereof at a concentration of about 2% to 8% by weight, (ii) liquid oil
at a
concentration of about 10% to 20% by weight, (iii) one or more phospholipids
at a
concentration of about 10% to 20% by weight, and (iv) dextrose at a
concentration
50% to 80% by weight.
. The lyophilized clarithromycin formulation may be reconstituted with
an appropriate amount of an aqueous solution (e.g., water, a buffer solution,
or an
aqueous solution with other additives such as tonicity modifiers) to re-form
an oil-
in-water emulsion that comprises clarithromycin or a pharmaceutically
acceptable
salt or ester thereof at a pharmaceutically effective concentration. In
certain
embodiments, the re-formed oil-in-water emulsion is physically, chemically, or
both
physically and chemically stable for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9,
10, 11, or
12 days under appropriate storage condition (e.g., at 2-8 C).
In certain embodiments, the re-formed oil-in-water clarithromycin
emulsion is non-vein irritating.
In certain embodiments, the re-formed oil-in-water clarithromycin
emulsion has a pH of about 6 to about 8 (e.g., about 7 or about 7.5).
In certain embodiments, the reconstituted oil-in-water emulsions
have the same chemical and physical characteristics as described above for the
lyophilizable oil-in-water emulsions. For example, in certain embodiments, the
reconstituted oil-in-water emulsion has an average diameter of oil droplets no
more
than about 200 nm, and/or a PFAT5 of less than about 0.05. In certain
embodiments, the reconstituted oil-in-water emulsion is isotonic and ready for
use.
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In certain embodiments, the reconstituted oil-in-water emulsion does not
further
comprise a compound that increases the amount of clarithromycin in the oil
droplets of the reconstituted emulsion. In certain embodiments, no more than
about 70%, 60%, 50%, 40%, 30%, or 20% of clarithromycin is present in the oil
droplets in the reconstituted oil-in-water emulsion.
The clarithromycin formulations of the present invention may be used
to treat or reduce the risk of infection for which clarithromycin is
effective. More
specifically, the lyophilizable clarithromycin oil-in-water formulations or
oil-in-water
emulsions reconstituted from lyophilized formulations may be administered to a
subject (e.g., human or other mammals) in need thereof at a pharmaceutically
effective amount by various routes, including but not limited to, intravenous,
intramuscular, intra-arterial, intrathecal, intraocular, subcutaneous, intra-
articular,
oral, topical, intravaginal and intra-peritoneal administration.
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EXAMPLES
EXAMPLE 1
PREPARATION OF LYOPHILIZABLE AND LYOPHILIZED VANCOMYCIN EMULSIONS
A 1000 g batch of vancomycin emulsion was prepared in the
following composition:
Component Su tiers % w/w /1000
Vancomycin hydrochloride, USP Hospira Inc. 2 20
Soy lecithin, EP American lecithin 4 40
com an
Medium chain tri I ceride, USP Sasol 1.5 15
Soybean oil, USP Corda 2.5 25
Dextrose, USP Ro uette 20 200
Water, de-ionized to add to 100 1000
The emulsion was prepared in the following steps:
(1) Weighed out and combined soy lecithin, medium chain
triglyceride and soybean oil in a compounding vessel;
(2) Added an appropriate amount of ethanol to dissolve all
components to obtain a clear solution;
(3) Applied vacuum to remove ethanol until the residual ethanol is
less than 1% the dry weight to obtain a clear oil solution;
(4) Added vancomycin HCI, dextrose and water in another
container, mixed well to dissolve the solids to obtain a clear aqueous
solution;
(5) Added the aqueous solution to the oil solution and applied a
high-shear homogenizer, e.g., an IKA Ultra Turrax mixer, to obtain a primary
emulsion (oil-in-water);
(6) Adjusted pH of the primary emulsion to 7.4 0.1;
(7) Passed the primary emulsion through a high-pressure
homogenizer (e.g., Microfluidizer 110L operating a 100 psi inlet air pressure)
for 3-
5 passes to obtain a sub-micron emulsion with average droplet size of about 77
nanometers;
(8) Passed the emulsion through a 0.2-micron sterilizing filter
(Nalgene) to obtain the lyophilizable emulsion;
(9) Filled 25 g of the sterilized emulsion into each 50 mL sterile
vial (500 mg vancomycin HCI per vial); and
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(10) Lyophiled using a shelf freeze-dryer (e.g., Advantage model
by Virtis) to obtain the lyophilized vancomycin emulsion.
The lyophilized vancomycin emulsion was tested as follow:
Lyophile appearance
The lyophilized emulsion in the vials was observed as off-white,
uniform cake-like, porous mass.
Reconstitution
Each vial of lyophilized emulsion was reconstituted with 15.4 g water
with gentle mixing for about 1-5 minutes to obtain a white, uniform, and
translucent
to opaque emulsion ("reformed emulsion").
The reformed emulsion
The reformed emulsion was observed under an optical microscope at
400x magnification, no crystals, visible droplets, or solid matter was
observed. pH
of the reformed emulsion was measured to be 7.1. Average droplet size of the
reformed emulsion was determined by a laser light scattering spectrometer
(Model
770 by Particle Sizing Systems) to be 82 nm_ The amount and integrity of
vancomycin HCI in the reformed emulsion was confirmed by HPLC.
Stability of the reformed emulsion
The reformed emulsion was diluted to 10 mg/mL with water and
further diluted to 1 mg/mL with 5% dextrose solution and tested for stability
for 7
days. At -20 C, 5 C and 25 C, the I mg/mL diluted emulsion was found to be
stable for at least 1 days and the 10 mg/mL emulsion stable for at least 7
days.
Vein irritation
The reformed vancomycin emulsion was tested for vein irritation
using rabbit marginal ear vein model by slow infusion at 5 mg/mL and 10 mUkg
daily for 7 consecutive days. Appearance and histopathological examinations
did
not reveal any abnormality in vein tissues compared to a negative control (5%
dextrose solution). It was concluded that the vancomycin emulsion was non-vein
irritating.
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EXAMPLE 2
PREPARATION OF LYOPHILIZABLE AND LYOPHILIZED CLARITHROMYCIN EMULSIONS
A 1000 g batch of vancomycin emulsion was prepared in the
following composition:
Component Suppliers % w/w /1000
Clarithromycin, USP Teva 1.5 15
Soy lecithin, EP American lecithin 4 40
com an y
Medium chain tri t ceride, USP Sasol 1.5 15
Soybean oil, USP Corda 2.5 25
Dextrose, USP Ro uette 20 200
Water, de-ionized to add to 100 1000
The emulsion was prepared in the following steps:
(1) Weighed out and combined soy lecithin, medium chain
triglyceride and soybeari oil in a compounding vessel;
(2) Added an appropriate amount of ethanol to dissolve all
components to obtain a clear solution;
(3) Applied vacuum to remove ethanol until the residual ethanol is
less than 1 !o the dry weight to obtain a clear oil solution;
(4) Added clarithromycin, dextrose and water in another
container, mixed well and added hydrochloric acid to about pH 2.3 to dissolve
the
solids to obtain a clear aqueous solution;
(5) Added the aqueous solution to the oil solution and applied a
high-shear homogenizer, e.g., an IKA Ultra Turrax mixer, to obtain a primary
emulsion;
(6) Adjusted pH of the primary emulsion to 6.0 = 6.2;
(7) Passed the primary emulsion through a high-pressure
homogenizer (e.g., Microfluidizer 110L operating a 100 psi inlet air pressure)
for 5
passes to obtain a sub-micron emulsion with average droplet size of about 170
nanometers;
(8) Passed the emulsion through a 0.2-micron sterilizing filter
(Milfipak-20 by Millipore) to obtain the lyophilizable emulsion;
(9) Filled 16.7 g of the sterilized emulsion into each 50 mL sterile
vial (250 mg clarithromycin per vial); and
(10) Lyophiled using a shelf freeze-dryer (e.g., Advantage model
by Virtis) to obtain the lyophilized clarithromycin emulsion.
The lyophilized clarithromycin emulsion was tested as follow:
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Lyophile appearance
The lyophilized emulsion in the vials was observed as off-white,
uniform cotton-candy-like porous mass.
Reconstitution
Each vial of lyophilized emulsion was reconstituted with 11.78 g
water with gentle mixing for about 2-5 minutes to obtain a white, uniform, and
opaque emulsion ("reformed emulsion").
The reformed emulsion
The reformed emulsion was observed under an optical microscope at
400x magnification, no crystals, visible droplets, or solid matter was
observed.
pH of the emulsion was measured to be 6.4_
Average droplet size of the emulsion was determined by a laser light
scattering spectrometer (Model zetasizer by Malvern Instruments) to be 157-180
nm.
The amount of clarithromycin in the reformed emulsion was
confirmed by HPLC.
EXAMPLE 3
DEVELOPMENT OF FREEZE-STABLE, ISOTONIC, AND
SUB-MICRON VANCOMYCIN EMULStONS
A I g batch of each vancomycin emulsion was prepared in the
following composition:
Mg/g F30 F31 F32 F33 F34 F35 F36 F37
Vancomycin HCI 5.16* 5.16 5.16 5.16 5.16 5.16 5.16 5.16
Soy lecithin, EP 10 10 10 10 10 10 10 10.00
Soybean oil, USP 6.25 6.25 6.25 6.25 6.25 6.25 6.25 6.25
Medium chain triglyceride, USP 3.75 3.75 3.75 3.75 3.75 3.75 3.75 3.75
Dextrose, USP 50 60 70 80 90 100 50 60
Glycerin, USP 4.5
Deionized water qs to 1000 1000 1000 1000 1000 1000 1000 1000
pH 4 4 4 4 4 4 4 7.00
Total oil 10 10 10 10 10 10 10 10
Lecithin to oil ratio 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1
* Equivalent to 5.0 mg/g vancomycin free base
The emulsion was prepared in the following steps:
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(1) Weighed out and combined soy lecithin, soybean oil and
medium chain triglyceride in a suitable container;
(2) Added an appropriate amount of ethanol to dissolve all
components to obtain a clear solution;
(3) Applied vacuum to remove ethanol until the residual ethanol is
less than 2% the dry weight to obtain a clear oil solution;
(4) Added vancomycin HCI, dextrose, glycerin and water in
another container, mixed well to dissolve the solids to obtain a clear aqueous
solution;
(5) Added the aqueous solution to the oil solution;
(6) Agitated the mixture of aqueous and oil solutions vigorously
using mini-beadbeater for 10 seconds to form a primary emulsion;
(7) Recorded pH and adjusted pH to 4.0 +/- 0.2 or 7.0 +/- 0.2 with
0.1 N HCI/NaOH if needed;
(8) Agitated the primary emulsion for 200 seconds using the same
mini-beadbeater to obtain a final emulsion;
(9) Passed the emulsion through a 0.2-micron filter (Spin-X); and
(10) Froze the filtered emulsion at - 20 C.
The frozen emulsions were tested as follow:
pH and Osmotic Pressure
Formulation Osmotic pressure (mOsm) pH
F30 261.8 3.80
F31 314.5 4.00
F32 357.0 3.95
F33 415.7 3.90
F34 471.8 3.94
F35 522.3 3.96
F36 325.6 4.07
F37 315.0 6.98
Average Droplet Diameter by Laser Light Scattering (nm) by a Malevrn
Zetasizer Particle Sizing Instrument
1.5 da s 6 da s
Formulation initiai - 20 C (one - 20 C (two
freeze-thaw 5 C 25 C freeze-thaw 5 C 25 C
c cle c cles
F30 143 141 141 143 149 138 146
F31 166 149 153 168 173 150 151
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F32 140 135 191 173 137 140 147
F33 218 146 149 150 222 158 149
F34 184 149 160 155 157 146 155
F35 155 152 152 157 151 161 152
F36 160 157 158 160 150 151 160
F37 147 143 145 157 141 159 141
The above results show that: (1) dextrose concentration at about 5% and about
6% provided iso-osmotic emulsion (F30, F31, F36 and F37); (2) F30, F31, F36
and
F37 also showed no significant change in droplet size after two cycles of
freeze-
thaw; and (3) it appears that sub-micron emulsions can be prepared with about
0.5 fo vancomycin, about 1% oil, about 1% lecithin, about 5% or about 6%
dextrose
at either pH 4 or 7, and such emulsions are freeze-stable.
EXAMPLE 4
DEVELOPMENT OF FREEZE-STABLE, ISOTONIC, AND SUB-MICRON VANCOMYCIN
EMULSIONS
Another set of vancomycin emulsions were prepared in the following
composition:
rng/g F-30 F38 F39 F40 F41 F42 F43 F44 F45 F46 F47 F48 F49
Vancomycin HCI 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16
5.16
Soy lecithin, EP 10 20 30 50 20 30 50 20 30 50 20.0 30 50
Soybean oil, USP 6.25 12.5 18.75 31.25 12.5 18.75 31.25 12.5 18.75 31.25 12.5
18.75 31.25
MCT, USP 3.75 7.5 11.2518.75 7.5 11.2518.75 7.5 11.2518.75 7.5 11.2518.75
Dextrose, USP 50 50 50 50 60 60 60 50 50 50 60 60 60
Glycerin, USP 4.5 4.5 4.5
Deionized water qs to 10001000 1000 1000 1000 1000 1000 1000 1000 1000 1000
1000 1000
pH 4 4 4 4 4 4 4 4 4 7 7 7
Total oil 10 20 30 50 20 30 50 20 30 50 20 30 50
Lecithin to oilratio 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1 1:1
The emulsion was prepared and tested according the procedure and
methods described in Example 3.
Average Droplet Diameter by Laser Light Scattering (nm)
2-day 2-day 2-day
-20 C (2 freeze- 5 C
Sample Name Time-0 thaw c cles 25 C
F-30 133.7 138.7 135.7 137.0
F-38 91.5 96.1 90.0 94.7
F-39 105.3 89.4 82.6 85.2
F-40 112.0 108.3 92.7 105.7
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F-41 99.0 87.5 136.0 100.0
F-42 133.3 107.7 92.6 107.0
F-43 106.3 108.3 93.3 109.0
F-44 104.3 108.0 93.4 106.0
F-45 107.0 108.0 93.8 107.3
F-46 112.3 113.0 99.1 111.0
F-47 106.0 109.0 97.4 107.3
F-48 90.7 93.1 87.1 92.0
F-49 93.4 130.7 87.6 94.3
The results show that formulations at a lecithin to oil ratio of 1:1 with
about 1% to about 5% oil, about 1% to about 5% soy lecithin, and about 5% to
about 6% dextrose, or about 5% dextrose with about 0.45% glycerol at pH 4 or 7
formed sub-micron emulsions that are freeze-stable.
EXAMPLE 5
DEVELOPMENT OF FREEZE-STABLE, 1SOTONIC, AND SUB-MICRON VANCOMYCIN
EMULSIONS
Another set of vancomycin emulsions were prepared in the following
composition:
mg/g F-50 F-51 F-52 F-53 F-54 F-55 F-56 F-57 F-58 F-59 F-60 F-61
Vancomycin HCI 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16 5.16
Egg lecithin 12 12 12 12
Soy lecithin 12 12 12 12
by Lipoid
Soy lecithin 12 12 12 12
b Phos holi id Co.
Soybean oil, USP 100 100 100 75 75 75 50 50 50 50 50 50
Dextrose, USP 50 50 50 50 50 50 50 50 50 50 50 50
Deionized H2O qst 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000
pH 4 4 4 4 4 4 4 4 4 7 7 7
Total oil 100 100 100 75 75 75 50 50 50 50 50 50
Lecithin to oil ratio 0.12:1 0.12:1 0.12:1 0.16:1 0.16:1 0.16:1 0.24:1 0.24:1
0.24:1 0.24:1 0.24:1 0.24:1
The emulsion was prepared and tested according the procedure and
methods described in Example 3.
Appearance and Microscopic Observation for Visible Droplets
Formulation Appearance Microscopic observation
F50 No emulsion formed
F51 No emulsion formed
F52 No emulsion formed
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F53 Coarse emulsion A lot visible dro s>5 m
F54 Coarse emulsion A lot visible dro s>5 m
F55 Coarse emulsion A lot visible dro s>5 m
F56 Coarse emulsion A lot visible dro s>5 1m
F57 Coarse emulsion A lot visible dro s>5 m
F58 Coarse emulsion A lot visible dro s>5 m
F59 Coarse emulsion A lot visible dro s>5 m
F60 Coarse emulsion A lot visible dro s>5 m
F61 Coarse emulsion A lot visible dro s>5 m
The results show that no sub-micron emulsion was formed with about
0.5% vancomycin, about 1.2% lecithin, and about 10% soybean oil using the
method described in Example 3. The compositions tested in this study are
similar
to the conventional fat emulsion, i.e., with oil of about 5% to about 10% and
lecithin
about 1.2 fo. Their failure to form a sub-micron emulsion suggested that high
oil
concentration (> about 5%) does not favor the formation of sub-micron
emulsion,
and the lecithin to oil ratio of greater than 0.24:1 is required for the
formation for
sub-micron emulsion.
EXAMPLE 6
DEVELOPMENT OF FREEZE-STABLE, ISOTONIC, AND SUB-MICRON VANCOMYCIN
EMULSIONS
A 16 g batch of each vancomycin emulsion was prepared according
to following compositions:
Mg/g F-62 F-63 F-64 F-65
Vancomycin HCI 5.16 5.16 5.16 5.16
Egg lecithin, EP 8.26 12.38 16.51 20.12
Soybean oil, USP 50 100 100 100
Dextrose, USP 50 50 50 50
Deionized water gs to 1000 1000 1000 1000
pH 4 4 4 4
Total oil 50 100 100 100
Lecithin to oil ratio 0.17:1 0.12:1 0.17:1 0.20:1
The emulsion was prepared in the following steps:
(1) Weighed out and combined egg lecithin and soybean oil in a
suitable container;
(2) Added an appropriate amount of ethanol to dissolve all
components to obtain a clear solution;
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(3) Applied vacuum to remove ethanol until the residual ethanol is
less than 2% the dry weight to obtain a clear oil solution;
(4) Added vancomycin HCI, dextrose and water in another
container, mixed well to dissolve the solids to obtain a clear aqueous
solution;
(5) Added the aqueous solution to the oil solution;
(6) Recorded pH and adjusted pH to 4.0 +/- 0.2 with 0.1 N
HCI/NaOH if needed;
(7) Agitated the mixture of aqueous and oil solutions by hand
shaking;
(8) Recorded pH and adjusted pH to 4.0 +/- 0.2 or 7.0 +/- 0.2 with
0.1 N HCI/NaOH if needed;
(9) Applied a high-shear homogenizer, e.g., an IKA Ultra Turrax
mixer, to obtain a primary emulsion (oil-in-water);
(10) Passed the primary emulsion through a high-pressure
homogenizer (e.g., Microfluidizer 110L operating at 100 psi inlet air
pressure) for 3-
5 passes to obtain a sub-micron emulsion.
(11) Passed the emulsion through a 0.2-micron sterilizing filter
(Sartoruis, CE); and
(12) Froze the sterilized emulsion at - 20 C.
The frozen emulsions were tested as follow:
Averade Droplet Diameter by Laser Light Scattering (nm)
-20 C, 1 day
Formulation fg Before filtration (One freeze- ~ day'
thaw c cle
F62 192.5 171 185 216
F63 186.8 186 195 184
F64 191 177 208 184
F65 167 164 178 162
Observation under microscope for visible droplets (>5 m)
-20 C, 1 day 250
Formulation filtration After filtration (One freeze- 1 day
thaw c cle
F62 No No Yes Yes
F63 Yes Yes Yes Yes
F64 Yes Yes Yes Yes
F65 Yes Yes Yes Yes
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This study demonstrated that it is possible to produce a sub-micron
emulsion with about 5% oil at lecithin-to-oil ratio of 0.17:1 (F62) using the
preparation method described in Example 6. However, this emulsion was not
freeze-stable.
The presence of large droplets (> 5 m) demonstrated that a high %
of oil (about 10%) accompanied with a low lecithin to oil ratio (0.12:1,
0.17:1 or
0.2:1) did not favor the formation of a sub-micron emulsion.
EXAMPLE 7
DEVELOPMENT OF FREEZE-STABLE, ISOTONIC, AND SUB-MICRON VANCOMYCIN
EMULSIONS
A 16 g batch of each vancomycin emulsion was prepared according
to following compositions:
mg/g F66 F67 F68 F69 F70
Vancornycin HCI 5.16 5.16 5.16 5.16 5.16
Egg lecithin, EP 8.26 12.38 16.51 20.12
Soy lecithin, EP 12.38
Medium chain triglyceride, USP 25.0 50.0 50.0 50.0 50.0
Soybean oil, USP 25.0 50.0 50.0 50.0 50.0
Dextrose, USP 50.0 50.0 50.0 50.0 50.0
Deionized water gs to 1000 1000 1000 1000 1000
PH 4 4 4 4 4
Total oil 50 100 100 100 100
Lecithin to oil ratio 0.17:1 0.12:1 0.17:1 0.20:1 0.12:1
The emulsion was prepared and tested according the procedure and
methods described in Example 6.
Observation Under Microscope for Large Droplets (> 5 m)
-20 C, 1 day Q
Formulation Before at on After filtration (One freeze- ~ day'
thaw c cle
F66 Yes Yes Yes Yes
F67 Yes Yes Yes Yes
F68 Yes Yes Yes Yes
F69 Yes Yes Yes Yes
F70 Yes Yes Yes Yes
This study confirmed that an about 5 mg/mL vancomycin composition
with a high oil concentration (? about 5%) accompanied with a low lecithin-to-
oil
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ratio (< 0.2:1) does not favor formation of a sub-micron emulsion - for either
soybean oil alone or a combination of soybean oil and medium chain
triglyceride,
or for egg lecithin or soy lecithin.
EXAMPLE 8
DEVELOPMENT OF FREEZE-STABLE, ISOTONIC, AND SUB-MICRON VANCOMYCIN
EMULSIONS
A 16 g batch of each vancomycin emulsion was prepared according
to following compositions:
mg/g F-38 F-71 F-72 F-73
Vancomycin HCI 5.16 5.16 5.16 5.16
Soy lecithin, EP 20 20 20 20
Soybean oil, USP 12.5 18.8 25.0 10.0
Medium chain tri I ceride, USP 7.5 11.3 15.0 10.0
Dextrose, USP 50 50 50 50
Deionized water gs to 1000 1000 1000 1000
pH 4 4 4 4
Total oil 20 30 40 20
Lecithin to oil ratio 1:1 0.67:1 0.5:1 1:1
The emulsion was prepared and tested according the procedure and
methods described in Example 6.
pH
Formulation Time-0 - 20 C (One freeze-thaw cycle) 25 C
1 day 2 day 3 day 42 day 1 day 2 day 3 day
F38 3.99 3.97 4.05 4.05 4.04 4.05
F71 4.16 Not 4.12 4.11 4.15 Not 4.31 4.33
F72 4.08 tested 4.06 4.14 4.06 tested 4.2 4.2
F73 4.06 4.11 4.09 3.93 4_ 19 4.26
Average Droplet Diameter by Laser Light Scattering (nm)
Formulation Time-0 - 20 C (One freeze-thaw cycte) 25 C
I day 2 day 3 day 42 day I day" 2 day 3 day
F38 113 123 117 115 127 112 112 108
F71 190 130 141 149 129 128 131 140
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T F72 190 141 138 139 146 185 136 136
F73 127 116 121 124 136 114 127 116
Observation Under Microscope for Large Droplets (> 5 um)
Formulation Time-0 - 20 C (One freeze-thaw cycle) 25 C
1 day 2 day 3 day 42 day I day 2 day 3 day
F38 None None Few Few Few None Few Few
F71 None None Few Few Few None Few Few
F72 None None Few Few Few None Few Few
F73 None None Few Few Few None None None
Percentage of Fat Greater Than 5 Micrometers at Time-0 (i.e., PFAT5)
by Accusizer 380
(%) F-38 F-71 F-72 F-73
AVG 0.002 0.003 0.004 0.003
Std Dev 0.001 0.001 0.002 0.003
The results show that submicron and freeze-stable emulsions were
obtained with the oil concentration no more than about 4% and lecithin-to-oil
ratio
of greater than about 0.5:1 using microfluidzation process. No significant
change
in pH or particle size was observed for all four formulations after 3 days
storage at
either -20 C or 25 C. F38 and F73 appeared to be better formulations based
average droplet size and on the microscope observation where fewer large
droplets were seen in F38 and F73 than the others.
EXAMPLE 9
PREPARATION OF A FROZEN ISOTONIC CLARITHROMYCIN EMULSION
A 16 g batch of each vancomycin emulsion is prepared according to
following compositions:
mg/g F-74 F-75
Clarithromycin 5.0 5.0
Soy lecithin, EP 20 20
Soybean oil, USP 12.5 10.0
Medium chain tri I ceride, USP 7.5 10.0
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Dextrose, USP 50 50
Deionized water gs to 1000 1000
pH 7 7
Total oil 20 20
Lecithin to oil ratio 1:1 1:1
The emulsion is prepared in the following steps:
(1) Weigh out and combine soy lecithin, soybean oil and medium
chain triglyceride in a suitable container;
(2) Add an appropriate amount of ethanol to dissolve all
components to obtain a clear solution;
(3) Apply vacuum to remove ethanol until the residual ethanol is
less than 2% the dry weight to obtain a clear oil solution;
(4) Add clarithromycin, dextrose and water in another container,
mixed well to dissolve the solids to obtain a clear aqueous solution;
(5) Add the aqueous solution to the oil solution;
(6) Record pH and adjust pH to 7.0 +/- 2 with 0.1N HCI/NaOH if
needed;
(7) Agitate the mixture of aqueous and oil solutions by hand
shaking;
(8) Record pH and adjust pH to 7.0 +/- 2 with 0.1 N HCI/NaOH if
needed;
(9) Apply a high-shear homogenizer, e.g., an IKA Ultra Turrax
mixer, to obtain a primary emulsion (oil-in-water);
(10) Pass the primary emulsion through a high-pressure
homogenizer (e.g., Microfluidizer 110L operating at 100 psi inlet air
pressure) for 3-
5 passes to obtain a sub-micron emulsion;
(11) Pass the emulsion through a 0.2-micron sterilizing filter
(Sartoruis, CE); and
(12) Freeze the sterilized emulsion at - 20 C.
The various embodiments described above can be combined to
provide further embodiments. All of the U.S. patents, U.S. patent application
publications, U.S. patent applications, foreign patents, foreign patent
applications
and non-patent publications referred to in this specification are incorporated
herein
by reference, in their entirety. Aspects of the embodiments can be modified,
if
necessary to employ concepts of the various patents, applications and
publications
to provide yet further embodiments.
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These and other changes can be made to the embodiments in light
of the above-detailed description. In general, in the following claims, the
terms
used should not be construed to limit the claims to the specific embodiments
disclosed in the specification and the claims, but should be construed to
include all
possible embodiments along with the full scope of equivalents to which such
claims are entitled. Accordingly, the claims are not limited by the
disclosure.
56
