U.S. patent application number 12/341780 was filed with the patent office on 2009-11-26 for antibiotic formulations, unit doses, kits, and methods.
This patent application is currently assigned to Nektar Therapeutics. Invention is credited to Chatan Charan, Sarvajna Dwivedi.
Application Number | 20090288658 12/341780 |
Document ID | / |
Family ID | 37744353 |
Filed Date | 2009-11-26 |
United States Patent
Application |
20090288658 |
Kind Code |
A1 |
Charan; Chatan ; et
al. |
November 26, 2009 |
Antibiotic Formulations, Unit Doses, Kits, and Methods
Abstract
An aqueous or powder composition includes anti-gram-negative
antibiotic or salt thereof being present at an amount ranging from
about 100 mg/ml to about 200 mg/ml. Another aqueous or powder
composition includes anti-gram-positive antibiotic or salt thereof
being present at a concentration ranging from about 0.6 to about
0.9 of the water solubility limit, at 25.degree. C. and 1.0
atmosphere, of the anti-gram-positive antibiotic or salt thereof.
Other embodiments include unit doses, kits, and methods.
Inventors: |
Charan; Chatan; (San Jose,
CA) ; Dwivedi; Sarvajna; (Redwood City, CA) |
Correspondence
Address: |
NEKTAR THERAPEUTICS
201 INDUSTRIAL ROAD
SAN CARLOS
CA
94070
US
|
Assignee: |
Nektar Therapeutics
San Carlos
CA
|
Family ID: |
37744353 |
Appl. No.: |
12/341780 |
Filed: |
December 22, 2008 |
Related U.S. Patent Documents
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
|
|
11529128 |
Sep 28, 2006 |
|
|
|
12341780 |
|
|
|
|
60722564 |
Sep 29, 2005 |
|
|
|
Current U.S.
Class: |
128/200.14 ;
514/40; 536/16.8 |
Current CPC
Class: |
A61P 29/00 20180101;
A61M 16/04 20130101; A61J 1/065 20130101; A61K 9/0019 20130101;
A61K 9/19 20130101; A61P 43/00 20180101; A61J 1/1412 20130101; A61K
31/7034 20130101; A61M 11/001 20140204; A61P 31/04 20180101; A61P
11/08 20180101; A61P 31/00 20180101; A61M 2202/064 20130101; A61P
11/00 20180101; A61K 31/573 20130101; A61K 9/08 20130101; A61K
31/7036 20130101; A61K 9/0078 20130101 |
Class at
Publication: |
128/200.14 ;
514/40; 536/16.8 |
International
Class: |
A61M 11/00 20060101
A61M011/00; A61K 31/7036 20060101 A61K031/7036; C07G 11/00 20060101
C07G011/00 |
Claims
1. An aqueous composition for aerosolization comprising amikacin
being present at an amount from about 100 mg/ml to about 200
mg/ml.
2. The aqueous composition of claim 1, wherein the aqueous
composition consists essentially of amikacin and water.
3. The aqueous composition of claim 1, wherein the amount amikacin
is from about 110 mg/ml to about 150 mg/ml, or a potency from about
500 .mu.g/mg to about 1100 .mu.g/mg, or both.
4. The aqueous composition of claim 1, wherein the aqueous
composition has a pH from about 4 to about 6.
5. The aqueous composition of claim 1, wherein the aqueous
composition has an osmolality ranging from about 90 mOsmol/kg to
about 500 mOsmol/kg.
6. The aqueous composition of claim 1, wherein the aqueous
composition is preservative-free.
7. The aqueous composition of claim 1, further comprising an
additional active agent.
8. The aqueous composition of claim 1, further comprising an
additional active agent selected from an anti-inflammatory and a
bronchodilator.
9. The aqueous composition of claim 1, further comprising a
bronchodilator selected from .beta.-agonist, anti-muscarinic agent,
and steroid.
10. The aqueous composition of claim 1, further comprising
albuterol.
11. The aqueous composition of claim 1, further comprising an
osmolality adjuster.
12. The aqueous composition of claim 1, wherein no precipitate
forms in the aqueous composition when the aqueous composition is
stored for 1 year at 25.degree. C.
13. The aqueous composition of claim 1, further comprising an
anti-gram positive antibiotic.
14. An aqueous composition, consisting essentially of amikacin, a
bronchodilator, and water.
15. The aqueous composition of claim 14, wherein the amikacin is
present in an amount ranging from about 100 mg/ml to about 200
mg/ml.
16. The aqueous composition of claim 14, wherein the aqueous
composition has a pH from about 3 to about 7, and an osmolality
from about 90 mOsmol/kg to about 500 mOsmol/kg.
17. The aqueous composition of claim 14, wherein the bronchodilator
is present in an amount of at least about 1 mg/ml.
18. The aqueous composition of claim 17, wherein the bronchodilator
is selected from a .beta.-agonist, anti-muscarinic agent, and
steroid.
19. The aqueous composition of claim 18, wherein the bronchodilator
comprises albuterol or salt thereof.
20. A unit dose, comprising a container and an aqueous composition
comprising amikacin, wherein the composition is preservative
free.
21. The unit dose of claim 20, wherein the amikacin is present at a
concentration from about 90 mg/ml to about 200 mg/ml.
22. A kit, comprising: a first container containing a first aqueous
solution comprising a first antibiotic; and a second container
containing a second aqueous solution comprising a second
antibiotic, wherein the first antibiotic is amikacin and the second
antibiotic is the same or different, and wherein a concentration,
or an amount, or both of the first antibiotic is the same as, or
different from a concentration, or an amount, or both of the second
antibiotic.
23. The kit of claim 22, wherein an amount of the first aqueous
solution is from about 2 ml to about 5 ml, an amount of the second
aqueous solution is from about 5 ml to about 8 ml, a concentration
of first antibiotic is from about 100 mg/ml to about 120 mg/ml, and
a concentration of second antibiotic is from about 120 mg/ml to
about 140 mg/ml.
24. The kit of claim 23, further comprising an aerosol
introducer.
25. The kit of claim 28, further comprising a vibrating mesh
nebulizer.
26. A kit, comprising: a first container containing a first aqueous
solution comprising amikacin; and a second container containing a
second aqueous solution comprising an anti-gram-negative
antibiotic, wherein a concentration, an amount, or both, of the
amikacin of the first aqueous solution is different from a
concentration, or an amount, or both, of the anti-gram-negative
antibiotic of the second aqueous solution.
27. The kit of claim 26, further comprising an aerosol
introducer.
28. The kit of claim 27, further comprising a vibrating mesh
nebulizer.
29. A kit, comprising: a first container containing a first
antibiotic comprising liquid amikacin; and a second container
containing a second antibiotic or salt thereof, wherein the first
and second antibiotics are the same or different, and wherein a
concentration, or an amount, or both of the first antibiotic in the
first container is different from a concentration, or an amount, or
both, of the second antibiotic or salt thereof in the second
container.
30. The kit of claim 29, wherein the second antibiotic comprises a
liquid.
31. The kit of claim 29, wherein the second antibiotic comprises a
powder.
32. The kit of claim 30, wherein the liquid amikacin is
preservative-free.
33. The kit of claim 29, further comprising an aerosol
introducer.
34. The kit of claim 33, further comprising a vibrating mesh
nebulizer.
35. A method of administering an antibiotic formulation to a
patient in need thereof, comprising: aerosolizing a liquid amikacin
formulation to administer the antibiotic formulation to the
pulmonary system of the patient, wherein the antibiotic formulation
has a concentration of amikacin ranging from about 100 mg/ml to
about 200 mg/ml.
36. The method of claim 35, wherein the antibiotic formulation
consists essentially of amikacin and water.
37. The method of claim 35, wherein the aerosolizing comprises
forming droplets having a mass median aerodynamic diameter of less
than about 10 .mu.m.
38. The method of claim 37, wherein the aerosolizing comprises
aerosolizing the antibiotic formulation in a vibrating mesh
nebulizer.
39. The method of claim 35, wherein the pulmonary administration
comprises prophylactic treatment of ventilator associated
pneumonia.
40. A method of administering an antibiotic formulation to a
patient in need thereof, comprising: inserting a tube into a
trachea of a patient; and aerosolizing an antibiotic formulation to
administer the antibiotic formulation to the lungs of the patient,
wherein the antibiotic formulation consists essentially of amikacin
and water.
41. The method of claim 40, wherein the pulmonary administration
comprises prophylactic treatment of ventilator associated
pneumonia, hospital associated pneumonia, community acquired
pneumonia, or combinations thereof.
42. The method of claim 40, wherein the aerosolizing comprises
aerosolizing the antibiotic formulation in a vibrating mesh
nebulizer.
Description
[0001] This application is a continuation of U.S. application Ser.
No. 11/529,128, filed Sep. 28, 2006, which claims priority to U.S.
Provisional Application No. 60/722,564, filed Sep. 29, 2005, which
is incorporated herein by reference in its entirety.
BACKGROUND
[0002] 1. Field of the Invention
[0003] The present invention relates to anti-infective, such as
antibiotic formulations, unit doses, kits, and methods, and in
particular to aminoglycoside formulations, unit doses, kits, and
methods
[0004] 2. Background of the Invention
[0005] The need for effective therapeutic treatment of patients has
resulted in the development of a variety of pharmaceutical
formulation delivery techniques. One traditional technique involves
the oral delivery of a pharmaceutical formulation in the form of a
pill, capsule, elixir, or the like. However, oral delivery can in
some cases be undesirable. For example, many pharmaceutical
formulations may be degraded in the digestive tract before the body
can effectively absorb them. Inhaleable drug delivery, where a
patient orally or nasally inhales an aerosolized pharmaceutical
formulation to deliver the formulation to the patient's respiratory
tract, may also be effective and/or desirable. In one inhalation
technique, an aerosolized pharmaceutical formulation provides local
therapeutic treatment and/or prophylaxis to a portion of the
respiratory tract, such as the lungs, to treat respiratory diseases
such as asthma and emphysema and/or to treat local lung infections,
such as fungal infections and cystic fibrosis. In another
inhalation technique, a pharmaceutical formulation is delivered
deep within a patient's lungs where it may be absorbed into the
bloodstream for systemic delivery of the formulation throughout the
body. Many types of aerosolization devices exist including devices
comprising a pharmaceutical formulation stored in or with a
propellant, devices that aerosolize a powder, devices which use a
compressed gas or other mechanism to aerosolize a liquid
pharmaceutical formulation, and similar devices.
[0006] One known aerosolization device is commonly referred to as a
nebulizer. A nebulizer imparts energy into a liquid pharmaceutical
formulation to aerosolize the liquid, and to allow delivery to the
pulmonary system, e.g. the lungs, of a patient. A nebulizer
comprises a liquid delivery system, such as a container having a
reservoir that contains a liquid pharmaceutical formulation. The
liquid pharmaceutical formulation generally comprises an active
agent that is either in solution or suspended within a liquid
medium. In one type of nebulizer, generally referred to as a jet
nebulizer, compressed gas is forced through an orifice in the
container. The compressed gas forces liquid to be withdrawn through
a nozzle, and the withdrawn liquid mixes with the flowing gas to
form aerosol droplets. A cloud of droplets is then administered to
the patient's respiratory tract. In another type of nebulizer,
generally referred to as a vibrating mesh nebulizer, energy, such
as mechanical energy, vibrates a mesh. This vibration of the mesh
aerosolizes the liquid pharmaceutical formulation to create an
aerosol cloud that is administered to the patient's lungs. In still
another type of nebulizer, ultrasonic waves are generated to
directly vibrate and aerosolize the pharmaceutical formulation.
[0007] Nebulizers are often used to deliver (1) an aerosolized
pharmaceutical formulation to a hospitalized or non-ambulatory
patient; (2) large doses of aerosolized active agent; and/or (3) an
aerosolized pharmaceutical formulation to a child or other patient
unable to receive a dry powder or propellant based pharmaceutical
formulation.
[0008] Nebulizers are useful for delivering an aerosolized
pharmaceutical formulation to the respiratory tract of a patient
who is breathing under the assistance of a ventilator. But there
are problems associated with the introduction of aerosolized
pharmaceutical formulation into ventilator circuits. For example,
by introducing the aerosolized pharmaceutical formulation into the
inspiratory line of the ventilator, significant residence volume
exists between the point of introduction and the patient's lungs.
Accordingly, large amounts of aerosolized pharmaceutical
formulation are needed and much of the formulation is lost to the
exhalation line. This problem is exacerbated when the nebulizer is
used in conjunction with ventilators having continual bias flows.
In addition, the large residence volume in the ventilator line may
dilute the aerosolized pharmaceutical formulation to an extent
where the amount delivered to the patient is difficult to reproduce
consistently.
[0009] U.S. Published Application Nos. 2004/0011358, 2004/0035490,
and 2004/0035413, which are incorporated herein by reference in
their entireties, disclose methods, devices, and formulations for
targeted endobronchial therapy. Aerosolized antibiotics are
delivered into a ventilator circuit. The aerosol generator, e.g.,
nebulizer, may be placed in the lower part of a Y-piece, for
example, distal to the Y, to be proximal to a patient airway and/or
endotracheal tube.
[0010] U.S. Pat. Nos. 5,508,269 and 6,890,907, which are
incorporated herein by reference in their entireties, disclose
aminoglycoside solutions for nebulization. The '269 patent
discloses that if the solution approaches the solubility of
tobramycin, 160 mg/ml, precipitation on storage is expected. The
'269 patent also discloses that a higher concentration of
tobramycin than is clinically needed is economically
disadvantageous. Further the '269 patent discloses that a more
concentrated solution will increase the osmolarity of the solution,
thus decreasing the output of the formulation with both jet and
ultrasonic nebulizers. The '269 patent discloses that the
alternative of a more concentrated solution in a smaller total
volume is also disadvantageous. The '269 patent further discloses
that most nebulizers have a dead space volume of 1 ml, i.e., that
of the last 1 ml of solution is wasted because the nebulizer is not
performing. Therefore, while for example, a 2 ml solution would
have 50% wastage, the 5 ml solution (the capacity of the nebulizer)
has only 20% wastage. Additionally, the '269 patent discloses that
since there is no sufficient aerosolization of the drug into the
small particles, the drug in large particles or as a solution is
deposited in the upper airways and induces cough and may also cause
bronchospasm. According to the '269 patent, large aerosol particles
also limit the drug delivery
[0011] There remains, however, a need for improved antibiotic
formulations, such as antibiotic formulations for nebulization.
There also remains a need for improved unit doses and kits of
antibiotic formulations. Accordingly, there also remains a need for
improved methods of making and/or using such antibiotic
formulations.
SUMMARY OF THE INVENTION
[0012] Accordingly, one or more embodiments of the present
invention satisfies one or more of these needs. Thus the present
invention provides antibiotic formulations, such as antibiotic
formulations for nebulization. The present invention also provides
unit doses and kits of antibiotic formulations. The present
invention further provides methods of making and/or using such
antibiotic formulations. Other features and advantages of the
present invention will be set forth in the description of invention
that follows, and will be apparent, in part, from the description
or may be learned by practice of the invention. The invention will
be realized and attained by the devices and methods particularly
pointed out in the written description and claims hereof.
[0013] In one aspect, one or more embodiments are directed to an
aqueous composition, comprising an antibiotic or salt thereof being
present at a therapeutic-effective (including
prophylatic-effective) amount. In one or more embodiments, the
therepautic-effective amount is based upon aerosolized
administration to the pulmonary system.
[0014] In one aspect, one or more embodiments are directed to an
aqueous composition, comprising anti-gram-negative antibiotic or
salt thereof being present at an amount ranging from about 90 mg/ml
to about 300 mg/ml.
[0015] In another aspect, an aqueous composition comprises
anti-gram-negative antibiotic or salt thereof, and optionally, a
bronchodilator.
[0016] In still another aspect, an aqueous composition comprises
anti-gram-positive antibiotic or salt thereof being present at a
concentration ranging from about 0.6 to about 0.9 of the water
solubility limit, at 25.degree. C. and 1.0 atmosphere, of the
anti-gram-positive antibiotic or salt thereof.
[0017] In yet another aspect, a unit dose comprises a container and
an aqueous composition, comprising anti-gram-negative antibiotic or
salt thereof being present at a concentration ranging from about
100 mg/ml to about 200 mg/ml.
[0018] In still another aspect, a kit comprises a first container
containing a first aqueous solution comprising an anti
gram-negative antibiotic or salt thereof; and a second container
containing a second aqueous solution comprising an anti
gram-negative antibiotic or salt thereof. A concentration, or an
amount, or both of the first aqueous solution is different from a
concentration, or an amount, or both, of the second aqueous
solution.
[0019] In yet another aspect, a kit comprises a first container
containing a first aqueous solution comprising anti gram-negative
antibiotic or salt thereof, and a second container containing a
second aqueous solution comprising anti gram-positive antibiotic or
salt thereof.
[0020] In another aspect, a unit dose comprises a container and a
powder comprising an antibiotic or salt thereof, wherein the powder
is present in an amount ranging from about 550 mg to about 900
mg.
[0021] In still another aspect, a unit dose comprises a container;
and a powder comprising an antibiotic or salt thereof, wherein the
powder is present in an amount ranging from about 150 mg to about
450 mg.
[0022] In yet another aspect, a kit comprises a first container
containing a first composition comprising anti-gram-positive or an
anti gram-negative antibiotic or salt thereof and a second
container containing a second composition comprising water. The
first composition and/or the second composition comprises
osmolality adjuster.
[0023] In another aspect, a kit comprises a first container
containing a powder comprising anti-gram-positive antibiotic or
salt thereof and a second container containing a powder comprising
anti gram-positive antibiotic or salt thereof. A concentration, or
an amount, or both, of the anti gram-positive antibiotic or salt
thereof in the first container is different from a concentration,
or an amount, or both, of the anti gram-positive antibiotic or salt
thereof in the second container.
[0024] In a further aspect, a kit comprises a first container
containing a solution comprising anti gram-negative antibiotic or
salt thereof and a second container containing a powder comprising
anti gram-positive antibiotic or salt thereof.
[0025] In still another aspect, a method of administering an
antibiotic formulation to a patient in need thereof comprises
aerosolizing an antibiotic formulation to administer the antibiotic
formulation to the lungs of the patient. The antibiotic formulation
has a concentration of antibiotic or salt thereof ranging from
about 90 mg/ml to about 300 mg/ml.
[0026] In another aspect, a method of administering an antibiotic
formulation to a patient in need thereof comprises inserting a tube
into a trachea of a patient. The method also comprises aerosolizing
an antibiotic formulation to administer the antibiotic formulation
to the lungs of the patient. The antibiotic formulation consists
essentially of anti-gram-negative antibiotic or salt thereof and
water.
[0027] In yet another aspect, a method of administering an
antibiotic formulation to a patient in need thereof comprises
aerosolizing an antibiotic formulation to administer the antibiotic
formulation to the lungs of the patient. The antibiotic formulation
comprises an antibiotic or salt thereof at a concentration ranging
from about 0.7 to about 0.9 of the water solubility limit, at
25.degree. C. and 1.0 atmosphere, of the antibiotic or salt
thereof.
[0028] In a further aspect, a method of administering an antibiotic
formulation to a patient in need thereof comprises dissolving an
antibiotic or salt thereof in a solvent to form an antibiotic
formulation, wherein the antibiotic or salt thereof is present at a
concentration ranging from about 0.6 to about 0.9 of the water
solubility limit, at 25.degree. C. and 1.0 atmosphere, of the
antibiotic or salt thereof. The method also includes aerosolizing
the antibiotic formulation to administer the antibiotic formulation
to the lungs of the patient.
[0029] In yet another aspect, a method of administering an
antibiotic formulation to a patient in need thereof comprises
dissolving an antibiotic or salt thereof in a solvent to form an
antibiotic formulation. The method also includes aerosolizing the
antibiotic formulation to administer the antibiotic formulation to
the lungs of the patient, wherein the aerosolizing is conducted
within about 16 hours of the dissolving.
[0030] In another aspect, a method involves forming a powder
comprising an antibiotic or salt thereof. The method includes
dissolving an antibiotic or salt thereof in a solvent to form a
solution having a concentration ranging from about 60 mg/ml to
about 120 mg/ml. The method also includes lyophilizing the solution
to form the powder.
[0031] In another aspect, a method involves forming a powder
comprising an antibiotic or salt thereof. The method comprises
dissolving an antibiotic or salt thereof in a solvent to form a
solution having a volume ranging from about 4.5 ml to about 5.5 ml.
The method also includes lyophilizing the solution to form the dry
powder.
[0032] In another aspect, any method which comprises forming a
powder may also include a method of reconstituting the powder to
form a liquid. Similarly any method which comprises forming a
liquid comprising an antibiotic (such as a solution) may also
include a method of removing the liquid to yield a powder.
[0033] In another aspect, any two or more of any of the foregoing
features, aspects versions or embodiments are combined.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] The present invention is further described in the
description of invention that follows, in reference to the noted
plurality of non-limiting drawings, wherein:
[0035] FIG. 1A illustrates components of a pulmonary drug delivery
system according to embodiments of the present invention.
[0036] FIG. 1B shows an embodiment of a device that can be used in
a pulmonary drug delivery system according to embodiments of the
invention.
[0037] FIG. 2A shows an exemplary off-ventilator configuration of a
pulmonary drug delivery system according to embodiments of the
invention.
[0038] FIG. 2B is a schematic view of a pharmaceutical delivery
device of one or more embodiments of the present invention, useful
for delivery of aerosolized medicaments.
[0039] FIG. 3 shows total drug recovered (nebulizer+filters) for
gentamicin as a function of fill mass and solution strength.
[0040] FIGS. 4a-b show emitted dose of gentamicin as a function of
solution strength and fill volume, after nebulization (FIG. 4a) for
15 minutes, and (FIG. 4) 30 minutes.
[0041] FIG. 5 shows gentamicin residual dose retained in a
nebulizer as a function of fill volume and solution strength.
[0042] FIG. 6 shows distribution of nebulized vancomycin (60 mg/ml
solution in normal saline) as a function of fill volume.
[0043] FIG. 7 shows emitted dose as a function of solution strength
and fill volume, for the case of vancomycin solution in 0.45%
saline.
[0044] FIG. 8 shows emitted dose as a function of solution strength
and fill volume, for the case of vancomycin solution in water for
injection (WFI).
[0045] FIG. 9 shows volume median diameter for nebulized gentamicin
as a function of solution strength and fill volume.
[0046] FIG. 10 shows cumulative particle size distributions for
gentamicin at different solution strengths and nebulizer fill
volumes.
[0047] FIG. 11 shows volume median diameter for nebulized
vancomycin (solution in WFI) as a function of solution strength and
fill volume.
[0048] FIG. 12 shows cumulative particle size distributions for
nebulized vancomycin (solution in WFI) at different solution
strengths and nebulizer fill volumes
[0049] FIG. 13 shows volume median diameter for nebulized
vancomycin (60 mg/ml solution in normal saline) as a function of
nebulizer fill volume.
[0050] FIG. 14 shows volume median diameter for nebulized
vancomycin (solution in 0.45% saline) as a function of solution
strength and fill volume.
[0051] FIG. 15 shows volume median diameter for antibiotic drug and
placebo solutions.
[0052] FIG. 16 is a graph showing amikacin stability over time (as
% related substance) for a formulation according to one or more
embodiments of the present invention, wherein the formulation was
stored at three different storage conditions.
DESCRIPTION OF THE INVENTION
[0053] Unless otherwise stated, a reference to a compound or
component includes the compound or component by itself, as well as
in combination with other compounds or components, such as mixtures
of compounds.
[0054] As used herein, the singular forms "a," "an," and "the"
include the plural reference unless the context clearly dictates
otherwise.
[0055] Reference herein to "one embodiment", "one version" or "one
aspect" shall include one or more such embodiments, versions or
aspects, unless otherwise clear from the context.
[0056] "Mass median diameter" or "MMD" is a measure of mean
particle size, since the powders of the invention are generally
polydisperse (i.e., consist of a range of particle sizes). MMD
values as reported herein are determined by centrifugal
sedimentation, although any number of commonly employed techniques
can be used for measuring mean particle size.
[0057] "Mass median aerodynamic diameter" or "MMAD" is a measure of
the aerodynamic size of a dispersed particle. The aerodynamic
diameter is used to describe an aerosolized powder in terms of its
settling behavior, and is the diameter of a unit density sphere
having the same settling velocity, generally in air, as the
particle. The aerodynamic diameter encompasses particle shape,
density and physical size of a particle. As used herein, MMAD
refers to the midpoint or median of the aerodynamic particle size
distribution of an aerosolized powder determined by cascade
impaction.
[0058] Anti-gram negative, and gram-negative antibiotic are used
interchangeably to refer to antibiotic active agents (and
formulations comprising such active agents) which have
effectiveness against gram negative bacteria. Similarly, anti-gram
positive, and gram-positive antibiotic are used interchangeably to
refer to antibiotic active agents (and formulations comprising such
active agents) which have effectiveness against gram positive
bacteria.
[0059] "Antibiotic" moreover includes anti-infectives, such as
antivirals and antifungals, as well as antibiotics, unless the
context indicates otherwise.
[0060] "Pharmaceutic formulation" and "composition" may be
sometimes used interchangeably to refer to a formulation comprising
an antibiotic.
[0061] As an overview, in one or more embodiments, an aqueous
composition comprises anti-gram-negative and/or anti-gram positive
antibiotic or salt thereof being present at an amount ranging from
about 100 mg/ml to about 200 mg/ml.
[0062] In one or more embodiments, an aqueous composition comprises
an antibiotic or salt thereof, and bronchodilator.
[0063] In one or more embodiments, an aqueous composition comprises
an antibiotic or salt thereof being present at a concentration
ranging from about 0.6 to about 0.9 of the water solubility limit,
at 25.degree. C. and 1.0 atmosphere, of the antibiotic or salt
thereof.
[0064] In one or more embodiments, a unit dose comprises a
container and an aqueous composition, comprising an
anti-gram-negative antibiotic or salt thereof at a concentration
ranging from about 100 mg/ml to about 200 mg/ml.
[0065] In one or more embodiments, a kit comprises a first
container containing a first aqueous solution comprising
anti-gram-negative antibiotic or salt thereof; and a second
container containing a second aqueous solution comprising
anti-gram-negative antibiotic or salt thereof. A concentration, or
an amount, or both, of the first aqueous solution is different from
a concentration, or an amount, or both, of the second aqueous
solution.
[0066] In one or more embodiments, a kit comprises a first
container containing a first aqueous solution comprising
anti-gram-negative antibiotic or salt thereof, and a second
container containing a second aqueous solution comprising
anti-gram-positive antibiotic or salt thereof.
[0067] In one or more embodiments, a unit dose comprises a
container and a powder comprising an antibiotic or salt thereof,
wherein the powder is present in an amount ranging from about 550
mg to about 900 mg.
[0068] In one or more embodiments, a unit dose comprises a
container; and a powder comprising an antibiotic or salt thereof,
wherein the powder is present in an amount ranging from about 150
mg to about 450 mg.
[0069] In one or more embodiments, a kit comprises a first
container containing a first composition comprising an
anti-gram-positive or an anti gram-negative antibiotic or salt
thereof and a second container containing a second composition
comprising water. The first composition and/or the second
composition comprises an osmolality adjuster.
[0070] In one or more embodiments, a kit comprises a first
container containing a powder comprising an anti-gram-positive
antibiotic or salt thereof and a second container containing a
powder comprising an anti-gram-positive antibiotic or salt thereof.
A concentration, or an amount, or both, of the anti-gram-positive
antibiotic or salt thereof in the first container is different from
a concentration, or an amount, or both, of the anti-gram-positive
antibiotic or salt thereof in the second container.
[0071] In one or more embodiments, a kit comprises a first
container containing a solution comprising an anti-gram-negative
antibiotic or salt thereof and a second container containing a
powder comprising anti-gram-positive antibiotic or salt
thereof.
[0072] In one or more embodiments, a method of administering an
antibiotic formulation to a patient in need thereof comprises
aerosolizing an antibiotic formulation to administer the antibiotic
formulation to the pulmonary system of the patient. The antibiotic
formulation has a concentration of anti-gram-negative antibiotic or
salt thereof ranging from about 100 mg/ml to about 200 mg/ml.
[0073] In one or more embodiments, a method of administering an
antibiotic formulation to a patient in need thereof comprises
inserting a tube into a trachea of a patient. The method also
comprises aerosolizing an antibiotic formulation to administer the
antibiotic formulation to the pulmonary system of the patient. The
antibiotic formulation consists essentially of an
anti-gram-negative antibiotic or salt thereof and water.
[0074] In one or more embodiments, a method of administering an
antibiotic formulation to a patient in need thereof comprises
aerosolizing an antibiotic formulation to administer the antibiotic
formulation to the pulmonary system of the patient. The antibiotic
formulation comprises an anti-gram-positive antibiotic or salt
thereof at a concentration ranging from about 0.7 to about 0.9 of
the water solubility limit, at 25.degree. C. and 1.0 atmosphere, of
the anti-gram-positive antibiotic or salt thereof.
[0075] In one or more embodiments, a method of administering an
antibiotic formulation to a patient in need thereof comprises
aerosolizing an antibiotic formulation using a vibrating mesh
nebulizer, and administering the antibiotic formulation to the
pulmonary system of the patient via an endotracheal tube, wherein
the nebulizer is positioned in close proximity to the endotracheal
tube.
[0076] In one or more embodiments, a method of administering an
antibiotic formulation to a patient in need thereof comprises
dissolving an anti-gram-positive antibiotic or salt thereof in a
solvent to form an antibiotic formulation, wherein the
anti-gram-positive antibiotic or salt thereof is present at a
concentration ranging from about 0.6 to about 0.9 of the water
solubility limit, at 25.degree. C. and 1.0 atmosphere, of the
anti-gram-positive antibiotic or salt thereof. The method also
includes aerosolizing the antibiotic formulation to administer the
antibiotic formulation to the pulmonary system of the patient.
[0077] In one or more embodiments, a method of administering an
antibiotic formulation to a patient in need thereof comprises
dissolving an antibiotic or salt thereof in a solvent to form an
antibiotic formulation. The method also includes aerosolizing the
antibiotic formulation to administer the antibiotic formulation to
the pulmonary system of the patient, wherein the aerosolizing is
conducted within about 16 hours of the dissolving.
[0078] In one or more embodiments, a method involves forming a
powder comprising an antibiotic or salt thereof. The method
includes dissolving an antibiotic or salt thereof in a solvent to
form a solution having a concentration ranging from about 60 mg/ml
to about 120 mg/ml. The method also includes lyophilizing the
solution to form the powder.
[0079] In one or more embodiments, a method involves forming a
powder comprising an antibiotic or salt thereof. The method
comprises dissolving an antibiotic or salt thereof in a solvent to
form a solution having a volume ranging from about 4.5 ml to about
5.5 ml. The method also includes lyophilizing the solution to form
the dry powder.
[0080] Therefore, in one or more embodiments, the present invention
involves concentrated antibiotic formulations. The antibiotic
formulations may comprise an aqueous composition of antibiotic or
salt thereof being present at a concentration ranging from about
0.6 to about 0.9, such as about 0.7 to about 0.8, of the water
solubility limit, at 25.degree. C. and 1.0 atmosphere, of the
antibiotic or salt thereof.
[0081] The concentration of the antibiotic, corrected for potency,
in one or more embodiments, may range from about 40 mg/ml to about
200 mg/ml, such as about 60 mg/ml to about 140 mg/ml, or about 80
mg/ml to about 120 mg/ml. For example, in the case of
anti-gram-negative antibiotics or salts thereof, the concentration
as corrected for potency may range from about 40 mg/ml to about 200
mg/ml, such as from about 90 mg/ml to about 200 mg/ml, about 110
mg/ml to about 150 mg/ml, or about 120 mg/ml to about 140 mg/ml. As
another example, in the case of anti-gram-positive antibiotics or
salts thereof, the concentration as corrected for potency may range
from about 60 mg/ml to about 140 mg/ml, such as about 80 mg/ml to
about 120 mg/ml.
[0082] The aqueous compositions typically have a pH that is
compatible with physiological administration, such as pulmonary
administration. For example, the aqueous composition may have a pH
ranging from about 3 to about 7, such as about 4 to about 6.
[0083] In addition, the aqueous compositions typically have an
osmolality that is compatible with physiological administration,
such as pulmonary administration. In one or more embodiments, the
aqueous composition may have an osmolality ranging from about 90
mOsmol/kg to about 500 mOsmol/kg, such as 120 mOsmol/kg to about
500 mOsmol/kg, or about 150 mOsmol/kg to about 300 mOsmol/kg.
[0084] In one or more embodiments, the aqueous compositions are
stable. For instance, in some cases, no precipitate forms in the
aqueous composition when the aqueous composition is stored for 1
year, or even 2 years, at 25.degree. C.
[0085] The potency of the antibiotic or salt thereof may range from
about 500 .mu.g/mg to about 1100 .mu.g/mg. In one or more
embodiments, the potency of anti-gram-negative antibiotics or salts
thereof, such as gentamicin, typically ranges from about 500
.mu.g/mg to about 1100 .mu.g/mg, such as about 600 .mu.g/mg to
about 1000 .mu.g/mg, or about 700 .mu.g/mg to about 800 .mu.g/mg.
The potency of anti-gram-positive antibiotics or salts thereof,
such as vancomycin, typically ranges from about 900 .mu.g/mg to
about 1100 .mu.g/mg, such as from about 950 .mu.g/mg to about 1050
.mu.g/mg.
[0086] The chromatographic purity level of the antibiotic or salt
thereof typically greater than about 80%, such as greater than
about 85%, greater than about 90%, or greater than about 95%. In
this regard, there is generally no major impurity greater than
about 10%, such as no greater than about 5% or no greater than
about 2%. For instance, the amount of heavy metals is typically
less than about 0.005 wt %, such as less than about 0.004 wt %,
less than about 0.003 wt %, less than about 0.002 wt %, or less
than about 0.001 wt %.
[0087] In the case of gentamicin, the compositions typically have a
gentamicin C.sub.1 content ranging from about 25% to about 50%,
such as about 30% to about 55%, about 35% to about 50%, or about
40% to about 45%, based on the total amount of gentamicin. The
compositions typically have a gentamicin C.sub.1a content ranging
from about 10% to about 35%, such as about 15% to about 30%, about
20% to about 25%, based on the total amount of gentamicin. The
compositions typically have a gentamicin C.sub.2 and C.sub.2a
content ranging from about 25 wt % to about 55 wt %, such as about
30% to about 50%, about 30% to about 45%, or about 35% to about
40%, based on the total amount of gentamicin.
[0088] In embodiments of the present invention comprising amikacin,
the compositions typically have an amikacin content ranging from
about 25% to about 50%, such as about 30% to about 55%, about 35%
to about 50%, or about 40% to about 45%, based on the total amount
of amikacin.
[0089] Nearly any anti-gram-negative, anti-gram-positive
antibiotic, or combinations thereof may be used. Additionally,
antibiotics may comprise those having broad spectrum effectiveness,
or mixed spectrum effectiveness. Antifungals, such as polyene
materials, in particular, amphotericin B are also suitable for use
herein. Examples of anti-gram-negative antibiotics or salts thereof
include, but are not limited to, aminoglycosides or salts thereof.
Examples of aminoglycosides or salts thereof include gentamicin,
amikacin, kanamycin, streptomycin, neomycin, netilmicin, paramecin,
tobramycin, salts thereof, and combinations thereof. For instance,
gentamicin sulfate is the sulfate salt, or a mixture of such salts,
of the antibiotic substances produced by the growth of
Micromonospora purpurea. Gentamicin sulfate, USP, may be obtained
from Fujian Fukang Pharmaceutical Co., LTD, Fuzhou, China. Amikacin
is typically supplied as a sulfate salt, and can be obtained, for
example, from Bristol-Myers Squibb. Amikacin may include related
substances such as kanamicin.
[0090] Examples of anti-gram-positive antibiotics or salts thereof
include, but are not limited to, macrolides or salts thereof.
Examples of macrolides or salts thereof include, but are not
limited to, vancomycin, erythromycin, clarithromycin, azithromycin,
salts thereof, and combinations thereof. For instance, vancomycin
hydrochloride is a hydrochloride salt of vancomycin, an antibiotic
produced by certain strains of Amycolatopsis orientalis, previously
designated Streptomyces orientalis. Vancomycin hydrochloride is a
mixture of related substances consisting principally of the
monohydrochloride of vancomycin B. Like all glycopeptide
antibiotics, vancomycin hydrochloride contains a central core
heptapeptide. Vancomycin hydrochloride, USP, may be obtained from
Alpharma, Copenhagen, Denmark.
[0091] In some embodiments, the composition comprises an antibiotic
and one or more additional active agents. The additional active
agent described herein includes an agent, drug, or compound, which
provides some pharmacologic, often beneficial, effect. This
includes foods, food supplements, nutrients, drugs, vaccines,
vitamins, and other beneficial agents. As used herein, the terms
further include any physiologically or pharmacologically active
substance that produces a localized or systemic effect in a
patient. An active agent for incorporation in the pharmaceutical
formulation described herein may be an inorganic or an organic
compound, including, without limitation, drugs which act on: the
peripheral nerves, adrenergic receptors, cholinergic receptors, the
skeletal muscles, the cardiovascular system, smooth muscles, the
blood circulatory system, synoptic sites, neuroeffector junctional
sites, endocrine and hormone systems, the immunological system, the
reproductive system, the skeletal system, autacoid systems, the
alimentary and excretory systems, the histamine system, and the
central nervous system.
[0092] Examples of additional active agents include, but are not
limited to, anti-inflammatory agents, bronchodilators, and
combinations thereof.
[0093] Examples of bronchodilators include, but are not limited to,
.beta.-agonists, anti-muscarinic agents, steroids, and combinations
thereof. For instance, the steroid may comprise albuterol, such as
albuterol sulfate.
[0094] Active agents may comprise, for example, hypnotics and
sedatives, psychic energizers, tranquilizers, respiratory drugs,
anticonvulsants, muscle relaxants, antiparkinson agents (dopamine
antagnonists), analgesics, anti-inflammatories, antianxiety drugs
(anxiolytics), appetite suppressants, antimigraine agents, muscle
contractants, additional anti-infectives (antivirals, antifungals,
vaccines) antiarthritics, antimalarials, antiemetics, anepileptics,
cytokines, growth factors, anti-cancer agents, antithrombotic
agents, antihypertensives, cardiovascular drugs, antiarrhythmics,
antioxicants, anti-asthma agents, hormonal agents including
contraceptives, sympathomimetics, diuretics, lipid regulating
agents, antiandrogenic agents, antiparasitics, anticoagulants,
neoplastics, antineoplastics, hypoglycemics, nutritional agents and
supplements, growth supplements, antienteritis agents, vaccines,
antibodies, diagnostic agents, and contrasting agents. The active
agent, when administered by inhalation, may act locally or
systemically.
[0095] The active agent may fall into one of a number of structural
classes, including but not limited to small molecules, peptides,
polypeptides, proteins, polysaccharides, steroids, proteins capable
of eliciting physiological effects, nucleotides, oligonucleotides,
polynucleotides, fats, electrolytes, and the like.
[0096] Examples of active agents suitable for use in this invention
include but are not limited to one or more of calcitonin,
amphotericin B, erythropoietin (EPO), Factor VIII, Factor IX,
ceredase, cerezyme, cyclosporin, granulocyte colony stimulating
factor (GCSF), thrombopoietin (TPO), alpha-1 proteinase inhibitor,
elcatonin, granulocyte macrophage colony stimulating factor
(GMCSF), growth hormone, human growth hormone (HGH), growth hormone
releasing hormone (GHRH), heparin, low molecular weight heparin
(LMWH), interferon alpha, interferon beta, interferon gamma,
interleukin-1 receptor, interleukin-2, interleukin-1 receptor
antagonist, interleukin-3, interleukin-4, interleukin-6,
luteinizing hormone releasing hormone (LHRH), factor IX, insulin,
pro-insulin, insulin analogues (e.g., mono-acylated insulin as
described in U.S. Pat. No. 5,922,675, which is incorporated herein
by reference in its entirety), amylin, C-peptide, somatostatin,
somatostatin analogs including octreotide, vasopressin, follicle
stimulating hormone (FSH), insulin-like growth factor (IGF),
insulintropin, macrophage colony stimulating factor (M-CSF), nerve
growth factor (NGF), tissue growth factors, keratinocyte growth
factor (KGF), glial growth factor (GGF), tumor necrosis factor
(TNF), endothelial growth factors, parathyroid hormone (PTH),
glucagon-like peptide thymosin alpha 1, IIb/IIIa inhibitor, alpha-1
antitrypsin, phosphodiesterase (PDE) compounds, VLA-4 inhibitors,
bisphosphonates, respiratory syncytial virus antibody, cystic
fibrosis transmembrane regulator (CFTR) gene, deoxyreibonuclease
(Dnase), bactericidal/permeability increasing protein (BPI),
anti-CMV antibody, 1 3-cis retinoic acid, oleandomycin,
troleandomycin, roxithromycin, clarithromycin, davercin,
azithromycin, flurithromycin, dirithromycin, josamycin, spiromycin,
midecamycin, leucomycin, miocamycin, rokitamycin, andazithromycin,
and swinolide A; fluoroquinolones such as ciprofloxacin, ofloxacin,
levofloxacin, trovafloxacin, alatrofloxacin, moxifloxicin,
norfloxacin, enoxacin, grepafloxacin, gatifloxacin, lomefloxacin,
sparfloxacin, temafloxacin, pefloxacin, amifloxacin, fleroxacin,
tosufloxacin, prulifloxacin, irloxacin, pazufloxacin,
clinafloxacin, and sitafloxacin, teicoplanin, rampolanin,
mideplanin, colistin, daptomycin, gramicidin, colistimethate,
polymixins such as polymixin B, capreomycin, bacitracin, penems;
penicillins including penicllinase-sensitive agents like penicillin
G, penicillin V, penicillinase-resistant agents like methicillin,
oxacillin, cloxacillin, dicloxacillin, floxacillin, nafcillin; gram
negative microorganism active agents like ampicillin, amoxicillin,
and hetacillin, cillin, and galampicillin; antipseudomonal
penicillins like carbenicillin, ticarcillin, azlocillin,
mezlocillin, and piperacillin; cephalosporins like cefpodoxime,
cefprozil, ceftbuten, ceftizoxime, ceftriaxone, cephalothin,
cephapirin, cephalexin, cephradrine, cefoxitin, cefamandole,
cefazolin, cephaloridine, cefaclor, cefadroxil, cephaloglycin,
cefuroxime, ceforanide, cefotaxime, cefatrizine, cephacetrile,
cefepime, cefixime, cefonicid, cefoperazone, cefotetan,
cefinetazole, ceftazidime, loracarbef, and moxalactam, monobactams
like aztreonam; and carbapenems such as imipenem, meropenem,
pentamidine isethiouate, lidocaine, metaproterenol sulfate,
beclomethasone diprepionate, triamcinolone acetamide, budesonide
acetonide, fluticasone, ipratropium bromide, flunisolide, cromolyn
sodium, ergotamine tartrate and where applicable, analogues,
agonists, antagonists, inhibitors, and pharmaceutically acceptable
salt forms of the above. In reference to peptides and proteins, the
invention is intended to encompass synthetic, native, glycosylated,
unglycosylated, pegylated forms, and biologically active fragments,
derivatives, and analogs thereof.
[0097] Active agents for use in the invention further include
nucleic acids, as bare nucleic acid molecules, vectors, associated
viral particles, plasmid DNA or RNA or other nucleic acid
constructions of a type suitable for transfection or transformation
of cells, i.e., suitable for gene therapy including antisense.
Further, an active agent may comprise live attenuated or killed
viruses suitable for use as vaccines. Other useful drugs include
those listed within the Physician's Desk Reference (most recent
edition), which is incorporated herein by reference in its
entirety.
[0098] The amount of antibiotic or other active agent in the
pharmaceutical formulation will be that amount necessary to deliver
a therapeutically or prophylactically effective amount of the
active agent per unit dose to achieve the desired result. In
practice, this will vary widely depending upon the particular
agent, its activity, the severity of the condition to be treated,
the patient population, dosing requirements, and the desired
therapeutic effect. The composition will generally contain anywhere
from about 1 wt % to about 99 wt %, such as from about 2 wt % to
about 95 wt %, or from about 5 wt % to 85 wt %, of the active
agent, and will also depend upon the relative amounts of additives
contained in the composition. The compositions of the invention are
particularly useful for active agents that are delivered in doses
of from 0.001 mg/day to 100 mg/day, such as in doses from 0.01
mg/day to 75 mg/day, or in doses from 0.10 mg/day to 50 mg/day. It
is to be understood that more than one active agent may be
incorporated into the formulations described herein and that the
use of the term "agent" in no way excludes the use of two or more
such agents.
[0099] Generally, the compositions are free of excessive
excipients. In one or more embodiments, the aqueous composition
consists essentially of the anti-gram-negative antibiotic, such as
amikacin, or gentamicin or both, and/or salts thereof and
water.
[0100] Further, in one or more embodiments, the aqueous composition
is preservative-free. In this regard, the aqueous composition may
be methylparaben-free and/or propylparaben-free. Still further, the
aqueous composition may be saline-free.
[0101] In one or more embodiments, the compositions comprise an
anti-infective and an excipient. The compositions may comprise a
pharmaceutically acceptable excipient or carrier which may be taken
into the lungs with no significant adverse toxicological effects to
the subject, and particularly to the lungs of the subject. In
addition to the active agent, a pharmaceutical formulation may
optionally include one or more pharmaceutical excipients which are
suitable for pulmonary administration. These excipients, if
present, are generally present in the composition in amounts
sufficient to perform their intended function, such as stability,
surface modification, enhancing effectiveness or delivery of the
composition or the like. Thus if present, excipient may range from
about 0.01 wt % to about 95 wt %, such as from about 0.5 wt % to
about 80 wt %, from about 1 wt % to about 60 wt %. Preferably, such
excipients will, in part, serve to further improve the features of
the active agent composition, for example by providing more
efficient and reproducible delivery of the active agent and/or
facilitating manufacturing. One or more excipients may also be
provided to serve as bulking agents when it is desired to reduce
the concentration of active agent in the formulation.
[0102] For instance, the compositions may include one or more
osmolality adjuster, such as sodium chloride. For instance, sodium
chloride may be added to solutions of vancomycin hydrochloride to
adjust the osmolality of the solution. In one or more embodiments,
an aqueous composition consists essentially of the
anti-gram-positive antibiotic, such as vancomycin hydrochloride,
the osmolality adjuster, and water.
[0103] Pharmaceutical excipients and additives useful in the
present pharmaceutical formulation include but are not limited to
amino acids, peptides, proteins, non-biological polymers,
biological polymers, carbohydrates, such as sugars, derivatized
sugars such as alditols, aldonic acids, esterified sugars, and
sugar polymers, which may be present singly or in combination.
[0104] Exemplary protein excipients include albumins such as human
serum albumin (HSA), recombinant human albumin (rHA), gelatin,
casein, hemoglobin, and the like. Suitable amino acids (outside of
the dileucyl-peptides of the invention), which may also function in
a buffering capacity, include alanine, glycine, arginine, betaine,
histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine,
isoleucine, valine, methionine, phenylalanine, aspartame, tyrosine,
tryptophan, and the like. Preferred are amino acids and
polypeptides that function as dispersing agents. Amino acids
falling into this category include hydrophobic amino acids such as
leucine, valine, isoleucine, tryptophan, alanine, methionine,
phenylalanine, tyrosine, histidine, and proline.
[0105] Carbohydrate excipients suitable for use in the invention
include, for example, monosaccharides such as fructose, maltose,
galactose, glucose, D-mannose, sorbose, and the like;
disaccharides, such as lactose, sucrose, trehalose, cellobiose, and
the like; polysaccharides, such as raffinose, melezitose,
maltodextrins, dextrans, starches, and the like; and alditols, such
as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol
(glucitol), pyranosyl sorbitol, myoinositol and the like.
[0106] The pharmaceutical formulation may also comprise a buffer or
a pH adjusting agent, typically a salt prepared from an organic
acid or base. Representative buffers comprise organic acid salts of
citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric
acid, succinic acid, acetic acid, or phthalic acid, Tris,
tromethamine hydrochloride, or phosphate buffers.
[0107] The pharmaceutical formulation may also include polymeric
excipients/additives, e.g., polyvinylpyrrolidones, celluloses and
derivatized celluloses such as hydroxymethylcellulose,
hydroxyethylcellulose, and hydroxypropylmethylcellulose, Ficolls (a
polymeric sugar), hydroxyethylstarch, dextrates (e.g.,
cyclodextrins, such as 2-hydroxypropyl-.beta.-cyclodextrin and
sulfobutylether-.beta.-cyclodextrin), polyethylene glycols, and
pectin.
[0108] The pharmaceutical formulation may further include flavoring
agents, taste-masking agents, inorganic salts (for example sodium
chloride), antimicrobial agents (for example benzalkonium
chloride), sweeteners, antioxidants, antistatic agents, surfactants
(for example polysorbates such as "TWEEN 20" and "TWEEN 80"),
sorbitan esters, lipids (for example phospholipids such as lecithin
and other phosphatidylcholines, phosphatidylethanolamines), fatty
acids and fatty esters, steroids (for example cholesterol), and
chelating agents (for example EDTA, zinc and other such suitable
cations). Other pharmaceutical excipients and/or additives suitable
for use in the compositions according to the invention are listed
in "Remington: The Science & Practice of Pharmacy", 19.sup.th
ed., Williams & Williams, (1995), and in the "Physician's Desk
Reference", 52.sup.nd ed., Medical Economics, Montvale, N.J.
(1998), both of which are incorporated herein by reference in their
entireties.
[0109] For MDI applications, the pharmaceutical formulation may
also be treated so that it has high stability. Several attempts
have dealt with improving suspension stability by increasing the
solubility of surface-active agents in the HFA propellants. To this
end U.S. Pat. No. 5,118,494, WO 91/11173 and WO 92/00107 disclose
the use of HFA soluble fluorinated surfactants to improve
suspension stability. Mixtures of HFA propellants with other
perfluorinated cosolvents have also been disclosed as in WO
91/04011. Other attempts at stabilization involved the inclusion of
nonfluorinated surfactants. In this respect, U.S. Pat. No.
5,492,688 discloses that some hydrophilic surfactants (with a
hydrophilic/lipophilic balance greater than or equal to 9.6) have
sufficient solubility in HFAs to stabilize medicament suspensions.
Increases in the solubility of conventional nonfluorinated MDI
surfactants (e.g. oleic acid, lecithin) can also reportedly be
achieved with the use of co-solvents such as alcohols, as set forth
in U.S. Pat. Nos. 5,683,677 and 5,605,674, as well as in WO
95/17195. A particularly useful class of MDIs are those which use
hydrofluoroalkane (HFA) propellants. The HFA propellants are
further particularly well suited to be used with stabilized
dispersions of an active agent such as formulations and composition
of aminoglycoside antibiotics. Suitable propellants, formulations,
dispersions, methods, devices and systems comprise those disclosed
in U.S. Pat. No. 6,309,623, the disclosure of which is incorporated
by reference in its entirety. All of the aforementioned references
being incorporated herein by reference in their entireties.
[0110] In one or more embodiments, the compositions comprise an
aerosol having a particle or droplet size selected to permit
penetration into the alveoli of the lungs, such as a mass median
aerodynamic diameter, less than about 10 .mu.m, less than about 7.5
.mu.m, less than about 5 m, and usually being in the range of about
0.1 .mu.m to about 5 .mu.m.
[0111] The compositions of the present invention may be made by any
of the various methods and techniques known and available to those
skilled in the art. In this regard, procedures such as lyophilizing
antibiotics to make powders and/or dissolving antibiotics in
solvents are known in the art.
[0112] For instance, a solution of antibiotic, e.g., amikacin
sulfate or gentamicin sulfate, may be made using the following
procedure. Typically, manufacturing equipment is sterilized before
use. A portion of the final volume, e.g., 70%, of solvent, e.g.,
water for injection, may be added into a suitable container.
Antibiotic or salt thereof may then be added. The antibiotic or
salt thereof may be mixed until dissolved. Additional solvent may
be added to make up the final batch volume. The batch may be
filtered, e.g., through a 0.2 .mu.m filter into a sterilized
receiving vessel. Filling components may be sterilized before use
in filling the batch into vials, e.g., 10 ml vials.
[0113] As an example, the above-noted sterilizing may include the
following. A 5 liter type 1 glass bottle and lid may be placed in
an autoclave bag and sterilized at elevated temperature, e.g.,
121.degree. C. for 15 minutes, using an autoclave. Similarly, vials
may be placed into suitable racks, inserted into an autoclave bag,
and sterilized at elevated temperature, e.g., 121.degree. C. for 15
minutes, using an autoclave. Also similarly, stoppers may be placed
in an autoclave bag and sterilized at elevated temperature, e.g.,
121.degree. C. for 15 minutes, using an autoclave. Before
sterilization, sterilizing filters may be attached to tubing, e.g.,
a 2 mm length of 7 mm.times.13 mm silicone tubing. A filling line
may be prepared by placed in an autoclave bag and sterilized at
elevated temperature, e.g., 121.degree. C. for 15 minutes, using an
autoclave.
[0114] The above-noted filtration may involve filtration into a
laminar flow work area. The receiving bottle and filters may be set
up in the laminar flow work area.
[0115] The above-noted filling may also be conducted under laminar
flow protection. The filling line may be unwrapped and placed into
the receiving bottle. The sterilized vials and stoppers may be
unwrapped under laminar flow protection. Each vial may be filled,
e.g., to a target fill of 5.940 g, and stoppered. A flip off collar
may be applied to each vial. The sealed vials may be inspected for
vial leakage, correct overseals, and cracks.
[0116] As another example, one or more antibiotics, e.g.,
vancomycin, gentamicin or amikacin, and/or a salt thereof, may be
prepared by lyophilizing the antibiotic to form a powder for
storage. The powder is then reconstituted prior to use. This
technique may be used when the antibiotic is unstable in
solution.
[0117] In one or more embodiments, the powder making process may
begin with forming a solution to be lyophilized. For example, an
antibiotic or salt thereof, such as amikacin, gentamicin or
vancomycin and/or salts thereof, may be dissolved in a solvent to
form a solution having an antibiotic concentration ranging from
about 80 mg/ml to about 150 mg/ml, such as about 90 mg/ml to about
130 mg/ml, or about 100 mg/ml to about 124 mg/ml. The solution to
be lyophilized may have a volume ranging from about 4.5 ml to about
5.5 ml, such as about 5 ml.
[0118] In other embodiments, the powder making process may begin
with forming a solution of an anti-gram negative antibiotic or salt
thereof, such as amikacin or salt thereof. The antibiotic and/or
salt may be dissolved in a solvent to form a solution having a
concentration ranging from about 80 mg/ml to about 130 mg/ml, such
as about 90 mg/ml to about 120 mg/ml, or about 100 mg/ml to about
110 mg/ml. The solution to be lyophilized may have a volume ranging
from about 4.5 ml to about 5.5 ml, such as about 5 ml.
[0119] The solvent for the solution to be lyophilized may comprise
water. The solution may be excipient-free. For instance, the
solution may be cryoprotectant-free.
[0120] In one or more embodiments, a suitable amount (e.g., 120 g
per liter of final solution) of drug substance (for example
vancomycin hydrochloride) may be dissolved, e.g., in about the 75%
of the theoretical total amount of water for injection under
nitrogen bubbling. The dissolution time may be recorded and
appearance may be evaluated.
[0121] Then, the dilution to the final volume with WFI may be
carried out. Final volume may be checked. Density, pH, endotoxin,
bioburden, and content by UV may be measured both before and after
sterile filtration.
[0122] The solution may be filtered before lyophilizing. For
instance, a double 0.22 .mu.m filtration may be performed before
filling. The filters may be tested for integrity and bubble point
before and after the filtration.
[0123] Pre-washed and autoclaved vials may be aseptically filled
using an automatic filling line to a target of 5 ml per vial and
then partially stoppered. In process check for fill volumes may be
done by checking the fill weight every 15 minutes.
[0124] The lyophilizing is generally conducted within about 72
hours, such as within about 8 hours, or within about 4 hours, of
the dissolving.
[0125] In one or more embodiments, the lyophilizing comprises
freezing the solution to form a frozen solution. The frozen
solution is typically held at an initial temperature ranging from
about -40.degree. C. to about -50.degree. C., such as about
-45.degree. C. During the initial temperature period, the pressure
around the frozen solution is typically atmospheric pressure. The
initial temperature period typically ranges from about 1 hour to
about 4 hours, such about 1.5 hours to about 3 hours, or about 2
hours.
[0126] The lyophilizing may further comprise raising a temperature
of the frozen solution to a first predetermined temperature, which
may range from about 10.degree. C. to about 20.degree. C., such as
about 15.degree. C. The time for the heat ramp from the initial
temperature to the first predetermined temperature generally ranges
from about 6 hours to about 10 hours, such as about 7 hours to
about 9 hours.
[0127] During the first predetermined temperature period, the
pressure around the solution typically ranges from about 100
.mu.bar to about 250 .mu.bar, such as about 150 .mu.bar to about
225 .mu.bar. The solution may be held at the first predetermined
temperature for a period ranging from about 20 hours to about 30
hours, such as from about 24 hours.
[0128] The lyophilizing may still further comprise raising a
temperature of the solution to a second predetermined temperature,
which may range from about 25.degree. C. to about 35.degree. C.,
such as about 30.degree. C. During the second predetermined
temperature period, the pressure around the frozen solution
typically ranges from about 100 .mu.bar to about 250 .mu.bar, such
as about 150 .mu.bar to about 225 .mu.bar. The solution may be held
at the second predetermined temperature for a period ranging from
about 10 hours to about 20 hours.
[0129] In view of the above, the lyophilization cycle may comprise
a freezing ramp, e.g., from 20.degree. C. to -45.degree. C. in 65
minutes, followed by a freeze soak, e.g., at -45.degree. C. for 2
hours. Primary drying may be accomplished with a heating ramp,
e.g., from -45.degree. C. to 15.degree. C. in 8 hours, followed by
a temperature hold, e.g., at 15.degree. C. for 24 hours at a
pressure of 200 .mu.bar. Secondary drying may be accomplished with
a heating ramp, e.g., from 15.degree. C. to 30.degree. C. in 15
minutes, followed by a temperature hold at 30.degree. C. for 15
hours at a pressure of 200 .mu.bar. At the end of the
lyophilization cycle, the vacuum may be broken with sterile
nitrogen, and the vials may be automatically stoppered.
[0130] The water content of the powder e.g., vancomycin powder, or
amikacin powder, is typically less than about 7 wt %, such as less
than about 5 wt %, less than about 4 wt %, less than about 3 wt %,
or less than about 2 or 1 wt %.
[0131] The chromatographic purity level of the powder, e.g.,
vancomycin powder, or amikacin powder, typically greater than about
80%, such as greater than about 90%, greater than about 95%, or
greater than about 97%. In this regard, there is generally no major
impurity greater than about 10%, such as no greater than about 7%
or no greater than about 5%. For instance, the amount of heavy
metals is typically less than about 0.005 wt %, such as less than
about 0.004 wt %, less than about 0.003 wt %, less than about 0.002
wt %, or less than about 0.001 wt %.
[0132] The powder is capable of being reconstituted with water at
25.degree. C. and 1.0 atmosphere and with manual agitation, in less
than about 60 seconds, such as less than about 30 seconds, less
than about 15 seconds, or less than about 10 seconds.
[0133] The powder typically has a large specific surface area that
facilitates reconstitution. The specific surface area typically
ranges from about 5 m.sup.2/g to about 20 m.sup.2/g, such as about
8 m.sup.2/g to 15 m.sup.2/g, or about 10 m.sup.2/g to 12
m.sup.2/g.
[0134] Upon reconstitution with water, the antibiotic solution
(such as vancomycin or amikacin) typically has a pH that ranges
from about 2.5 to about 7, such as about 3 to about 6. Amikacin in
particular may have a pH of about 5.5 to about 6.3.
[0135] In addition to use formulations for nebulization, the
formulations of the present invention may be administered other
routes, e.g., parenteral administration.
[0136] One or more embodiments involve methods for treating or
preventing pulmonary infections, including nosocomial infections,
in animals, including, especially, humans. The method generally
comprises administering to an animal subject or human patient in
need thereof, as an aerosol, a therapeutically or prophylactically
effective amount of the antibiotic or salt thereof. Several
antibiotics may be delivered in combination according to the
invention, or in seriatim. In one or more embodiments, the amounts
delivered to the airways, if delivered systemically in such
amounts, would not be sufficient to be therapeutically effective
and would certainly not be enough to induce toxicity. At the same
time, in such embodiments, such amounts can result in sputum levels
of antibiotic of more than about 10-100 times the minimum
inhibitory concentration ("MIC").
[0137] In one particular embodiment, the pharmaceutical formulation
comprises an antibiotic for administration to a ventilated patient
to treat or prevent ventilator associated pneumonia (VAP) and/or
hospital-acquired pneumonia (HAP) and/or community acquired
pneumonia (CAP) as well as other forms of pneumonia, and other
respiratory infections or conditions. Such administration is
described in U.S. patent application Ser. Nos. 10/430,658;
10/430,765; and 10/991,092, and in U.S. Provisional Application
Nos. 60/378,475; 60/380,783; 60/420,429; 60/439,894; 60/442,785;
60/682,099, and in U.S. Patent Application Publication No.
2005/021766, all of which are incorporated herein by reference in
their entireties.
[0138] In one aspect, the aerosolized particles are prevented from
undergoing significant hygroscopic enlargement, since particles
enrobed in water will tend to condense on the walls. For instance,
the method may involve reducing humidity in the ventilator circuit
by a predetermined amount before nebulization begins. In this
embodiment, the humidity may facilitate an MMAD of less than about
3 .mu.m or less than about 1.5 .mu.m. In another embodiment, each
aerosol particle is delivered enrobed in a substantially
anhygroscopic envelope.
[0139] Of course, embodiments can be used where diameters are
greater. Moreover, in some cases, the present invention
contemplates adjustments to the surface electrical charges on the
particles or the walls. For example, assuming surface charge on the
device is important, the present invention contemplates embodiments
wherein the components of the device connectors are made of metal
(or at least coated with metal). Alternatively, the components can
be treated with agents (e.g. wetting agents, detergents, soaps) to
adjust surface charge.
[0140] In one aspect, the method comprises inserting an aerosol
delivery end of the device within said patient's trachea to create
a positioned device. The antibiotic composition is aerosolized
under conditions such that the composition is delivered through
said aerosol delivery end of the device to the patient, wherein the
aerosol first contacts the patient's trachea (thereby bypassing the
oro-pharynx). The method may involve administering a mixture of
antibiotics and is particularly appropriate for intubated
patients.
[0141] In another aspect, a method of administering comprises
administering to free breathing patients by way of an aerosol
generator device and/or system for administration of aerosolized
medicaments such as those disclosed in U.S. Patent Application
Publication Nos. 20050235987, 20050211253, 20050211245,
20040035413, and 20040011358, the disclosures of which are
incorporated herein by reference in their entirities.
[0142] Such devices may deliver medicament phasically or
non-phasically. Additionally or alternatively, such devices may
incorporate a chamber or reservoir to accumulate and periodically
dispense the aerosolized medicament. In one or more embodiments, an
aerosolized medicament comprises amikacin.
[0143] In one or more embodiments, the method of administering an
antibiotic formulation involves dissolving an antibiotic or salt
thereof in a solvent to form an antibiotic formulation. The
aerosolizing is conducted within about 16 hours, such as with about
12 hours, or within about 8 hours, of the dissolving.
[0144] In another aspect, particular with respect to
"constant-flow" ventilators, the present invention contemplates
limiting the delivery event to the inspiratory phase of the
ventilator cycle and, if possible, at a reduced flow-rate. Thus, in
one embodiment, aerosolization is actuated during (or in fixed
relation to) the inspiration phase of the breathing cycle.
[0145] It is not intended that the present invention be limited to
particular dosages. On the other hand, the efficiency of the
aerosol systems and methods described herein permit amounts to be
delivered that are too low to be generally effective if
administered systemically, but are nonetheless effective amounts
when administered in a suitable and pharmaceutically acceptable
formulation directly to the airway. Importantly, while efficiencies
can be increased, in some embodiments efficiencies are not
increased at the expense of control over the dose. Thus, lower
efficiencies are contemplated as preferred when delivery is more
reproducible.
[0146] It is not intended that the present invention be limited to
antimicrobials that only kill particular organisms. The present
invention contemplates drugs and drug combinations that will
address a wide variety of organisms. In one or more embodiments,
the present invention contemplates drugs or drug combinations
effective in the treatment of infections caused by P. aeruginosa,
S. aureus, H. influenza, and S. pneumoniae and/or
antibiotic-resistant strains of bacteria such as
methicillin-resistant S. aureus, and Acetinobacter species, among
others.
[0147] Moreover, while certain embodiments of the present invention
are presented in the context of the intubated patient, other
patients at risk for infection are contemplated as treatable with
the compositions, methods, and devices of the present invention.
For example, the elderly (particularly those in nursing homes),
horses, dogs and cats in competitions (show and racing animals),
animals that frequently travel (e.g., circus animals), animals in
close quarters (e.g., zoos or farms), humans and animals in general
are at risk for lung infections. The present invention contemplates
delivery of aerosols to the trachea and/or deep lung for such
individuals--both prophylactically (i.e., before symptoms) and
under acute conditions (i.e., after symptoms)--wherein said
aerosols comprise antimicrobials, and in particular, the antibiotic
mixtures described above.
[0148] In one embodiment, the present invention contemplates
administering the appropriate medication to a patient diagnosed
with ARDS or chronic obstructive pulmonary disease (COPD).
[0149] One or more embodiments are directed to unit doses
comprising a container and the compositions.
[0150] Examples of the container include, but are not limited to,
vials, syringes, ampoules, and blow fill seal. For instance, the
vial may be a colorless Type I borosilicate glass ISO 6R 10 mL vial
with a chlorobutyl rubber siliconized stopper, and rip-off type
aluminum cap with colored plastic cover.
[0151] The amount of the composition in the unit dose typically
ranges from about 2 ml to about 15 ml, such as from about 3 ml to
about 10 ml, about 4 ml to about 8 ml, or about 5 ml to about 6
ml.
[0152] The amount of the antibiotic in the unit dose, adjusted for
potency, typically ranges from about 150 mg to about 900 mg, such
as about 400 mg to about 750 mg. For instance, an amount of the
anti-gram-negative antibiotic or salt thereof may range from about
400 mg to about 750 mg. As another example, the amount of
anti-gram-positive antibiotic or salt thereof may range from about
150 mg to about 450 mg, or from about 550 mg to about 900 mg.
[0153] One or more embodiments are directed to kits. For instance,
the kit may includes a first container containing a first aqueous
solution comprising anti-gram-negative antibiotic or salt thereof
and a second container containing a second aqueous solution
comprising anti-gram-negative antibiotic or salt thereof. A
concentration, or an amount, or both, of the first aqueous solution
is different from a concentration, or an amount, or both, of the
second aqueous solution. For instance, the amount of the first
aqueous solution may range from about 2 ml to about 5 ml, and the
amount of the second aqueous solution may range from about 5 ml to
about 8 ml.
[0154] In one or more embodiments, the kit includes a first
container containing a first aqueous solution comprising
anti-gram-negative antibiotic or salt thereof. A second container
contains a second aqueous solution comprising anti-gram-positive
antibiotic or salt thereof. The concentrations and/or amounts of
the anti-gram-negative antibiotic or salt and the
anti-gram-positive antibiotic or salt may be the same or
different.
[0155] In one or more embodiments, a kit includes a first container
containing a first composition comprising an antibiotic or salt
thereof. A second container contains a second composition
comprising water. The first composition and/or the second
composition comprises an osmolality adjuster.
[0156] In one or more embodiments, a kit includes a first container
containing a powder comprising anti-gram-positive antibiotic or
salt thereof. A second container contains a powder comprising
anti-gram-positive antibiotic or salt thereof. A concentration, or
an amount, or both of the anti-gram-positive antibiotic or salt
thereof in the first container is different from a concentration,
or an amount, or both of the anti-gram-positive antibiotic or salt
thereof in the second container.
[0157] For instance, the amount of the anti-gram-positive
antibiotic or salt thereof in the first container may range from
about 400 mg to 600 mg. The amount of the anti-gram-positive
antibiotic or salt thereof in the second container may range from
about 600 mg to about 800 mg.
[0158] In another aspect, a kit may include a first container
containing a solution comprising anti-gram-negative antibiotic or
salt thereof. A second container may contain a powder comprising
anti-gram-positive antibiotic or salt thereof. Alternatively, the
anti-gram-negative antibiotic or salt thereof may be a powder, and
the anti-gram-positive antibiotic or salt thereof may be a solution
or dispersion. An amount of the anti-gram-positive antibiotic or
salt thereof generally ranges from about 150 mg to about 900
mg.
[0159] The kits may further comprise a package, such as a bag, that
contains the first container and the second container.
[0160] The kits may further comprise an aerosolization apparatus.
The aerosolization apparatus may be of any type that is capable of
producing respirable particles or droplets. Alternatively, the
antibiotic may be dissolved in or suspended in a liquid propellant,
as described in U.S. Pat. Nos. 5,225,183; 5,681,545; 5,683,677;
5,474,759; 5,508,023; 6,309,623; or 5,655,520, all of which are
incorporated herein by reference in their entireties. In such
cases, the aerosolization apparatus may comprise a metered dose
inhaler (MDI).
[0161] Alternatively or additionally, the pharmaceutical
formulation may be in a liquid form and may be aerosolized using a
nebulizer as described in WO 2004/071368, which is herein
incorporated by reference in its entirety, as well as U.S.
Published Application Nos. 2004/0011358 and 2004/0035413, which are
both herein incorporated by reference in their entireties. Other
examples of nebulizers include, but are not limited to, the
Aeroneb.RTM.Go or Aeroneb.RTM.Pro nebulizers, available from
Aerogen, Inc. of Mountain View, Calif.; the PARI eFlow and other
PARI nebulizers available from PARI Respiratory Equipment, Inc. of
Midlothian, Va.; the Lumiscope.RTM. Nebulizer 6600 or 6610
available from Lumiscope Company, Inc. of East Brunswick, N.J.; and
the Omron NE-U22 available from Omron Healthcare, Inc. of Kyoto,
Japan.
[0162] It has been found that a nebulizer of the vibrating mesh
type, such as one that that forms droplets without the use of
compressed gas, such as the Aeroneb.RTM. Pro provides unexpected
improvement in dosing efficiency and consistency. By generating
fine droplets by using a vibrating perforated or unperforated
membrane, rather than by introducing compressed air, the
aerosolized pharmaceutical formulation can be introduced into the
ventilator circuit without substantially affecting the flow
characteristics within the circuit and without requiring a
substantial re-selection of the ventilator settings. In addition,
the generated droplets when using a nebulizer of this type are
introduced at a low velocity, thereby decreasing the likelihood of
the droplets being driven to an undesired region of the ventilator
circuit. Furthermore, the combination of a droplet forming
nebulizer and an aerosol introducer as described is beneficial in
that there is a reduction in the variability of dosing when the
ventilator uses different tidal volumes, thus making the system
more universal.
[0163] Using an adaptor, device or system as disclosed in U.S.
application Ser. No. 10/991,092 and/or U.S. Provisional Application
No. 60/682,099, and/or U.S. Application Publication No.
2005/0217666, all of which are incorporated herein by reference in
their entireties, in connection with the administration of
aerosolized antibiotics offers substantial benefits. For example,
when using such adaptors, substantially less pharmaceutical
formulation is lost to the environment which results in a reduction
in bacterial resistance against the antibiotic. In addition, the
adaptors, devices or systems are able to deliver a more consistent
dose which is particularly useful for antibiotic therapy.
[0164] FIG. 1A shows an embodiment of an adapter or system for
aerosol delivery of medicaments, comprising a pulmonary drug
delivery system ("PDDS") 100 suitable for use with the present
invention. The PDDS 100 may include a nebulizer 102 (also called an
aerosolizer), which aerosolizes a liquid medicament stored in
reservoir 104. The aerosol exiting nebulizer 102 may first enter
the T-adaptor 106 that couples the nebulizer 102 to the ventilator
circuit. The T-adaptor 106 is also coupled to the circuit wye 108
that has branching ventilator limbs 110 and 112.
[0165] Coupled to one of the ventilator limbs 110 or 112 may be an
air pressure feedback unit 114, which equalizes the pressure in the
limb with the air pressure feedback tubing 116 connected to the
control module 118. In the embodiment shown, feedback unit 114 has
a female connection end (e.g., an ISO 22 mm female fitting)
operable to receive ventilator limb 112, and a male connection end
(e.g., an ISO 22 mm male fitting) facing opposite, and operable to
be inserted into the ventilator. The feedback unit may also be
operable to receive a filter 115 that can trap particulates and
bacteria attempting to travel between the ventilator circuit and
tubing 116.
[0166] The control module 118 may monitor the pressure in the
ventilator limb via tubing 116, and use the information to control
the nebulizer 102 through system cable 120. In other embodiments
(not shown) the control module 118 may control aerosol generation
by transmitting wireless signals to a wireless control module on
the nebulizer 102.
[0167] During the inhalation phase of the patient's breathing
cycle, aerosolized medicament entering T-adaptor 106 may be mixed
with the respiratory gases from the inspiratory ventilator limb 112
flowing to the patient's nose and/or lungs. In the embodiment
shown, the aerosol and respiratory gases flow through nose piece
122 and into the nasal passages of the patient's respiratory
tract.
[0168] Other embodiments of the circuit wye 108 shown in FIG. 1A
are also contemplated in embodiments of the invention.
[0169] Referring to FIG. 1B, a nebulizer 85, which may have a top
portion 93 through which liquid may be provided may be incorporated
into a ventilator breathing circuit of a ventilated patient. The
breathing circuit may comprise a "Y" connector 88, which may in
turn have an inlet portion 89, an endotracheal tube portion 90 and
an outlet portion 91. The inlet portion 89 carries air provided
from the ventilator 92 toward the patient. The endotracheal tube
portion 90 of the Y connector 88 carries the incoming air to the
patient's respiratory tract; this direction is represented by arrow
"a". The endotracheal tube portion 90 also carries the patient's
exhalation to the outlet portion 91 of the Y connector 88, and the
outlet portion may lead to an exhaust, represented by arrow "b", to
remove the patient's exhalation from the system. The nebulizer 85
of the present invention aerosolization element generates an
aerosol cloud 94 that remains substantially within the inlet
portion 89 of the Y connector 88 when there is no inspiratory air
flowing through the inlet portion, by virtue of the aerosolization
element, as described above, producing a low velocity mist. In this
manner, aerosol that is generated when there is no inhalation air
being provided will not be carried out through the outlet portion
91 of the Y connector and lost to the ambient environment.
Accordingly, a dose of aerosolized medication may be preloaded,
i.e., produced and placed substantially within the inlet portion 89
prior to an inhalation phase being sent by the ventilator 92. In
this manner, such medication can be swept into a patient's
respiratory system at the very start of the inhalation cycle. This
may be of particular benefit in the case of neonatal patients and
in other instances in which only the initial blast of inhalation
phase will reach the target portion of the respiratory system. In
alternate embodiments, the ventilator may generate a continuous
bias flow of gas through the ventilator circuit. The bias flow may
push some of the aerosolized medicament through the outlet portion
91, but there is still an overall benefit from having the
aerosolized medicament preloaded through the ventilator
circuit.
[0170] Referring now to FIG. 2A, an embodiment of an off-ventilator
configuration of an adapter and/or system for pulmonary delivery is
shown. In FIG. 2A, the adapter 400 is intended for off-ventilator
use, and includes an endpiece 402 that is coupled to a nebulizer
404 and wye 406. The nebulizer 404 may include reservoir 408, which
supplies the liquid medicament that is aerosolized into connector
410. The connector 410 can provide a conduit for the aerosolized
medicament and gases to travel from the wye 406 to endpiece 402,
and then into the patient's mouth and/or nose. The first wye limb
412 may be connected to a pump or source of pressurized respiratory
gases (not shown), which flow through the wye limb 412 to the
endpiece 402. A one-way valve 413 may also be placed in the limb
412 to prevent respired gases from flowing back into the pump or
gas source. The limb 412 may also include a pressure feedback port
414 that may be connected to a gas pressure feedback unit (not
shown). In the embodiment shown, a feedback filter 416 may be
coupled between the port 414 and feedback unit.
[0171] The off-ventilator adapter 400 may also include a second wye
limb 420, which includes a filter 422 and one-way valve 424,
through which gases may pass during an exhalation cycle. The filter
422 may filter out aerosolized medicament and infectious agents
exhaled by the patient to prevent these materials from escaping
into the surrounding atmosphere. The one-way valve 424 can prevent
ambient air from flowing back into the adapter 400.
[0172] A general form of an aerosolized composition delivery system
1100 is shown in FIG. 2B. The aerosolized composition delivery
system 1100 delivers an aerosolized composition to a portion of a
user's respiratory tract, such as the user's lungs. The aerosolized
composition delivery system 1100 is useful in delivering the
aerosolized composition to a patient whose breathing is being
assisted by a ventilator 1105 but may also be configured to be used
to deliver a composition to a non-ventilated patient. The
ventilator circuit 1110 is shown diagrammatically in FIG. 2B.
Extending from the ventilator 1105 is an inhalation line 1115 and
an exhalation line 1120. The inhalation line 1115 and the
exhalation line 1120 are both composed of tubing having an airflow
lumen extending therethrough. The inhalation line 1115 and the
exhalation line 1120 meet at an adaptor 1145 remote from the
ventilator 1105. At the adapter 1145 the lumen of the inhalation
line 1115 is in communication with the lumen from the exhalation
line 1120, and both lumens are in communication with a patient line
1130. The patient line 1130 comprises a lumen that extends to the
lumen of an endotracheal or tracheotomy tube 1135, which is
inserted into a patient. The tube 1135 has an opposite end that may
extend into or near the lungs of the user. Accordingly, in use,
oxygenated air is introduced into the inhalation line 1115 by the
ventilator 1105. The oxygenated air passes through the lumen of the
inhalation line 1115, into the patient line 1130, through the lumen
of the tube 1135, and into the lungs of the patient. The patient
then exhales, either naturally or by applying negative pressure
from the ventilator, and the exhaled air passes through the tube
1135, through the patient line 1130, and through the exhalation
line 1120 to the ventilator 1105. The cycle is continuously
repeated to assist the patient's breathing or to entirely control
the breathing of the patient.
[0173] The adapter 1145 introduces aerosolized composition into the
ventilator circuit 1110. The aerosol that is introduced by the
adapter 1145 is generated by an aerosolization apparatus 1150,
which comprises a reservoir for containing a composition. Thus, in
one or more embodiments, aerosolization energy is supplied to the
aerosolization device by an energy source 1160 to generate the
aerosolized composition. The aerosolized pharmaceutical formulation
passes through a passage 1165 to the adapter 1145 where it may be
introduced into the ventilator circuit 1110. The aerosolization
apparatus 1150 may be, for example, a jet nebulizer where the
energy source is compressed air, a vibrating mesh nebulizer where
the energy source is wave energy, an ultrasonic nebulizer, or a
metered dose inhaler where the energy source is a propellant that
boils under ambient conditions.
[0174] Examples of the adaptor 1145 for introducing the aerosolized
pharmaceutical formulation are disclosed in U.S. application Ser.
No. 10/991,092, filed Nov. 17, 2004, and U.S. Provisional
Application No. 60/682,099, which applications are herein
incorporated by reference in their entirety.
[0175] The introduction of the aerosolized pharmaceutical
formulation at the adapter 1145 is advantageous in many respects
over systems where the aerosol is introduced into the inhalation
line 1115 or within the ventilator 1105. For example, by
introducing the aerosolized pharmaceutical formulation at the
adapter 1145, the ventilator circuit volume from the point of
introduction to the patient's lungs is substantially reduced.
Accordingly, the aerosolized pharmaceutical formulation is more
concentrated and is less diffused throughout the ventilator circuit
1110. In addition, if the formulation is added in the inhalation
line 1115, much of the formulation is drawn into the exhalation
line 1120, further limiting the efficiency of the administration.
Because of this diffusion and reduced efficiency, the consistency
of dosing is difficult to control in known systems. Also, the
presence of high quantities of the aerosolized pharmaceutical
formulation that are not administered to the lungs of the patient
may be undesirable in that much of the aerosol may be introduced
into the environment where it may be inhaled by healthcare workers
or others.
[0176] Therefore, the adaptor 1145 of the invention has been
designed to introduce the aerosolized pharmaceutical formulation in
an improved manner to increase the efficiency and/or the
consistency of the dosing. The adaptor 1145 serves to reduce the
amount of aerosolized pharmaceutical formulation that is drawn into
the exhalation line 1120 of the ventilator circuit 1120.
[0177] The adaptors of the present invention when used in a
ventilator circuit are often able to reproducibly and efficiently
deliver pharmaceutical formulation. For instance, the present
invention is typically able to reproduce the delivered dose within
about .+-.10%, .+-.8%, .+-.6%, .+-.4%, .+-.2%, or .+-.1%, of the
total nominal dose. The present invention is often able to achieve
a delivered efficiency of at least about 30%, such as at least
about 40%, at least about 50%, at least about 60%, at least about
70%, at least about 80%, or at least about 90%.
[0178] The adaptor of the present invention typically has minimal
impact on the patient to ventilator interface. The minimal impact
allows the ventilator to react more efficiently to the patient. The
adaptor and valves are arranged so that at an air flow rate of 60
L/min, the pressure drop between the first end and the second end
of the adaptor is often less than about 50 cm H.sub.2O, such as
less than about 30 cm H.sub.2O, less than about 5 cm H.sub.2O, less
than about 4 cm H.sub.2O, less than about 3 cm H.sub.2O, less than
about 2H.sub.2O, less than about 1 cm H.sub.2O, less than about 0.5
cm H.sub.2O, or less than about 0.1 cm H.sub.2O, and may range from
about 0.05 cm H.sub.2O to about 10 cm H.sub.2O, about 1 cm H.sub.2O
to about 5 cm H.sub.2O, or about 2 cm H.sub.2O to about 4 cm
H.sub.2O. At an air flow rate of 30 L/min, the pressure drop
between the first end and the second end of the adaptor is
typically ranges from about 1 cm H.sub.2O to about 2 cm
H.sub.2O.
[0179] The adaptor may be made of a transparent, translucent, or
opaque material. Using a transparent material is advantageous
because the user can visually inspect the functioning of the
adaptor. Examples of materials for the adaptor include, but are not
limited, to polymers, such as polypropylene, SAN (styrene
acrylonitrile copolymer), ABS (acrylonitrile-butadiene-styrene),
polycarbonate, acrylic polysulfone, K-resin.RTM.
styrene-butadiene-copolymer (available from Chevron Phillips
Chemical), polyethylene, PVC (polyvinyl chloride), polystyrene, and
the like.
[0180] For vibrating mesh nebulizers, such as the Aeroneb Pro and
the PARI eFlow, reproducible administrations can result from
smaller first channel volumes. It has been determined, for example,
that the first channel volume for an adaptor 1145 used with a
vibrating mesh nebulizer may be any volume greater than about 10
ml, such as from about 10 ml to about 1000 ml, about 50 ml to about
200 ml, or about 90 ml. Both the stored volume and valving affect
the performance of the present invention.
[0181] Additional examples of devices and methods are disclosed in
U.S. patent application Ser. No. 11/436,329, "Valves, Devices, and
Methods for Endobronchial Therapy," filed May 18, 2006, which is
incorporated herein by reference in its entirety.
[0182] The present invention is not limited to any precise desired
outcome when using the above-described compositions, devices, and
methods. However, it is believed that the compositions, devices,
and methods of the present invention may result in a reduction in
mortality rates of intubated patients, a decrease in the incidence
of resistance (or at least no increase in resistance) because of
the reduced systemic antibiotic exposure and elevated exposure at
the targeted mucosal surface of the lung caused by local
administration. As noted above, it is contemplated that the
compositions, devices, and methods of the present invention are
useful in the treatment of pneumonia (and may be more effective
than systemic treatment--or at the very least, a useful adjunct).
It is believed that related infections may also be prevented or
reduced (e.g., prevention of sepsis, suppression of urinary tract
infections, etc.)
[0183] Of course, a reduced use of systemic antibiotics because of
the efficacy of the compositions, devices, and methods of the
present invention may result in reduced cost, reduced time on IV
lines, and/or reduced time on central lines). Moreover, such a
reduction should reduce antibiotic toxicity (as measured by reduced
incidence of diarrhea and C. difficile infection, better nutrition,
etc.)
[0184] It is believed that the compositions, devices, and methods
of the present invention will locally result in a reduction of the
ET/Trach tube biofilm. This should, in turn, get rid of secretions,
decrease airway resistance, and/or decrease the work of breathing.
The latter should ease the process of weaning the patient off of
the ventilator.
[0185] The present invention contemplates specific embodiments that
can replace commonly used elements of a ventilator system. In one
or more embodiments, the present invention contemplates an adapter
attachable to a ventilator circuit and to an endotracheal tube,
wherein the adaptor comprises an aerosol generator. While not
limited to any precise desired outcome, it is contemplated that the
adapter with integral generator will reduce the effects of the
ventilator on all conventional aerosol systems (jet, ultrasonic and
MDI), and at the same time enhance the positive qualities of a
device like the Aerogen.TM. pro. Again, while not limited to any
precise desired outcome, it is contemplated that the adapter with
integral generator will (1) reduce variability in delivery (reduced
effects of humidification, bias flow, continuous vs
breath-actuated) so as to achieve the same delivery (no matter what
commercial ventilator system is used); (2) allow for maximal
effects of breath actuation; and (3) allow for maximal effect to
enhanced nebulizer efficiency using nebulizers having no dead
volume.
[0186] The present invention is not limited to the precise
configuration or nature of the circuit. In one embodiment, said
circuit is a closed circuit. In another embodiment, said circuit is
an open circuit.
[0187] Again, the present invention is not limited to particular
vent configurations. In one embodiment, said inspiratory and said
expiratory lines are connected to a mechanical ventilator. In one
embodiment, said mechanical ventilator controls a breathing cycle,
said cycle comprising an inspiration phase. In one embodiment, the
aerosol is administered during the inspiration phase of the
breathing cycle.
[0188] Although the present invention has been described in
considerable detail with regard to certain versions thereof, other
versions are possible, and alterations, permutations and
equivalents of the version shown will become apparent to those
skilled in the art upon a reading of the specification and study of
the drawings. For example, the relative positions of the elements
in the aerosolization device may be changed, and flexible parts may
be replaced by more rigid parts that are hinged, or otherwise
movable, to mimic the action of the flexible part. In addition, the
passageways need not necessarily be substantially linear, as shown
in the drawings, but may be curved or angled, for example. Also,
the various features of the versions herein can be combined in
various ways to provide additional versions of the present
invention. Furthermore, certain terminology has been used for the
purposes of descriptive clarity, and not to limit the present
invention. Therefore, any appended claims should not be limited to
the description of the preferred versions contained herein and
should include all such alterations, permutations, and equivalents
as fall within the true spirit and scope of the present
invention.
[0189] The foregoing description will be more fully understood with
reference to the following Examples. Such Examples, are, however,
merely representative of methods of practicing one or more
embodiments of the present invention and should not be read as
limiting the scope of the invention.
Example 1
[0190] This Example involves determining the solubility of
gentamicin sulfate in water and saline. The required strengths were
initially set at 20 mg/ml, 40 mg/ml, and up to 200 mg/ml.
[0191] Water Solubility Determinations
[0192] Solubility in water was determined via visual assessment.
Osmolality and pH were also determined.
[0193] The batch size of all the solutions manufactured for the
solubility determination studies was 10 ml. The method of
manufacture consisted of weighing the appropriate amount of
gentamicin sulfate and then taking to final volume with water. It
was noted that especially for higher concentrations, the solution
was first shaken by hand and then placed on a magnetic stirrer to
ensure complete dissolution.
[0194] Table 1 lists the pH and osmolality values obtained for
solutions of gentamicin sulfate in water for injection (WFI) with
concentrations ranging from 20 mg/ml to 400 mg/ml.
TABLE-US-00001 TABLE 1 Test Matrix for Gentamicin Solution in WFI
Active Weight of Gentamicin Concentration Sulfate
Dispensed.sup.(*.sup.) Osmolality (mg/ml) (mg/ml) pH (mOsmol/kg) 20
34 4.81 61 40 68 4.72 101 80 136 4.92 197 120 204 4.93 275 200 340
5.01 524 250 425 5.06 1178 300 510 5.13 2013 350 595 5.20 NR 400
680 5.26 NR .sup.(*.sup.)Activity of gentamicin sulfate = 58.8%, so
conversion factor = 1.701 NR: No result, sample did not freeze
[0195] As seen in Table 1, all the solutions had a pH that was
higher than 4, which is considered to be acceptable for drug
delivery to the lungs. However, with regard to osmolality readings,
doses greater than 200 mg/ml exceeded the targeted range.
[0196] Gentamicin Sulfate in 0.9% Saline Solution
[0197] The solubility, pH, and osmolality of gentamicin sulfate
solutions prepared with 0.9% saline solution were determined. The
solubility was determined by visual assessment. Only three
concentrations of gentamicin were investigated (20, 40, and 80
mg/ml).
[0198] Table 2 lists the parameters measured for gentamicin
solutions and the observations recorded during manufacture.
TABLE-US-00002 TABLE 2 Osmolality and pH of Gentamicin Sulfate in
0.9% Saline Active Weight of Gentamicin Concentration Sulfate
Dispensed Osmolality (mg/ml) (mg/ml) pH (mOsmol/kg) 20 34 4.94 318
40 68 4.72 353 80 136 4.82 445
Example 2
[0199] This Example involves developing the freeze-drying cycle for
the clinical manufacture of the Vancomycin HCl lyophilisate. A 120
mg, 240 mg, and 480 mg of Vancomycin HCl/vial strength were
investigated.
[0200] Materials/Equipment
[0201] Materials [0202] Vancomycin hydrochloride, USP,
Alpharma--Denmark [0203] ISO 6R clear type I glass vials, Nuova
Ompi--Italy [0204] 20 mm freeze-drying stoppers, West
Pharmaceutical Service--USA [0205] 20 mm flip-off caps, Capsulit
S.p.A.--Italy [0206] 13 mm freeze-drying stoppers, West
Pharmaceutical Service--USA [0207] 13 mm flip-off caps, West
Pharmaceutical Service--USA
[0208] Equipment [0209] Glassware for Vancomycin solution before
and after filtration (bottles). [0210] Pressure vessel,
Sartorius--Germany [0211] Balance to check the filling weight (10
mg sensitivity), Sartorius--Germany [0212] Digital pH meter,
Mettler Toledo--Switzerland [0213] Karl Fischer automatic titrator
DL38, Mettler Toledo--Switzerland. [0214] 0.22 .mu.m sterilizing
PVDF filter, Pall [0215] Manual doser, Hirschmann [0216] Isolator,
E.Co.Tec--Italy [0217] Lyophilizer, BOC Edwards Lyoflex 04 (or
Minifast 8000) with the following characteristics: 0.4 m2 (or 0.8
m2) shelf surface; temperature range -50.degree. C. to 50.degree.
C.; PT 100 temperature probes; Pirani gauge for vacuum monitoring;
coil condenser with ice capacity of 8 kg; condenser coil inlet
temperature to -60.degree. C., stainless steel trays with a
thickness of about 2 mm; semiautomatic crimping machine
(Flexseal--Denmark) [0218] DSC Pyris Diamond--USA
[0219] Composition
[0220] Solubility Study
[0221] The solubility of the Vancomycin HCl has been evaluated in
order to establish a suitable formulation to obtain a final
lyophilised product which matches all the criteria required by its
use as pharmaceutical form.
[0222] The solubility coupled with a pH evaluation of Vancomycin
HCl solutions at different concentration was the first step to
focus the suitable final formulation for a better development of
the lyophilization cycle.
[0223] A saturated solution of Vancomycin HCl in water for
injection was prepared by adding under stirring the active agent to
the solvent.
[0224] At first, the solution was clear with the solid suspended as
an agglomerate; after the solid worked as crystallization nucleus
and a new precipitation occurred; so the solutions became white and
more viscous because the solid partially swells.
[0225] Suspension was stirred for 48 h in order to reach the
equilibrium conditions for the dissolution.
[0226] Suspensions was filtered first through a paper filter and
then through a 0.45 .mu.m PVDF filter discarding the first drops of
solution which could have been diluted because of the binding of
the product to the membrane.
[0227] The resulting solution obtained after the two filtrations
was stored at 2-8.degree. C. in order to evaluate if precipitation
of the solid occurs.
[0228] The filtrated solutions of Vancomycin HCl in water coming
from the respective saturated solutions, was diluted to reach a
final concentration which gave an Abs value at k=280 nm included
into the calibration curve.
[0229] Each diluted solution was analysed in triplicate with UV at
.lamda.=280 nm. For each solution the absorbances have been
mediated and the final value has been substituted in the respective
calibration curve equation to calculate the concentration.
[0230] The maximum solubility of Vancomycin HCl in water is 140.9
mg/mL.
[0231] pH of Vancomycin HCl Solution in Water
[0232] Besides the solubility evaluation it was also measured the
pH and density of Vancomycin HCl solutions at different
concentrations which could have been taken into account for the
development of the formulation and of the lyophilization cycle.
TABLE-US-00003 Solution Concentration Density (mg/mL) pH (g/mL)
140.8 3.4-3.5 1.046 130.5 3.5-3.7 1.042 120.26 3.6-3.8 1.037 110.16
3.7-3.9 1.034 100.12 3.8-4.1 1.027
[0233] The pH varied within a restricted range for each
concentration and the overall pH within 140 mg/mL and 100 mg/mL was
stable around the acid value.
[0234] Formulation
[0235] Vancomycin hydrochloride was dissolved in water for
injection to form 100 mg/ml formulations in 1.00 ml and 1.20 ml
amounts, as shown below.
TABLE-US-00004 Quantity Ingredients Amount/ml Amount/Unit
Vancomycin HCl 100.00 120.00 mg Water for injection to 1.00 ml to
1.20 ml
[0236] Vancomycin hydrochloride was also dissolved in water for
injection to form 120 mg/ml formulations in 1.00 ml, 2.00 ml, and
4.00 ml amounts, as shown below.
TABLE-US-00005 Quantity Amount/Unit Ingredients Amount/ml 120
mg/vial 240 mg/vial 480 mg/vial Vancomycin 120.0 120.00 mg 240.00
mg 480.00 mg HCl Water for to 1.00 ml to 1.00 ml to 2.00 ml to 4.00
ml injection
[0237] DSC Studies
[0238] DSC was performed on the ready to fill solution with a
concentration of 100 mg/ml and 120 mg/ml.
[0239] The DSC runs were performed by cooling the samples to
-50.degree. C. at a cooling rate of 1.degree. C./min, and by
heating them back to 20.degree. C. at different scan rates after a
period of few minutes of isothermal step.
[0240] Samples amount ranged approximately from 1 to 3 mg.
[0241] All the peaks corresponding to the detected thermal events
were calculated as onset temperature.
[0242] The DSC studies showed that there was a main event of
crystallization during freezing and that there is no evidence of
smaller crystallization events. These phenomena seem to indicate an
absence of amorphous phase during freezing and a complete retention
of crystalline structure by vancomycin, as confirmed by the lack of
glass transitions events during the heating steps in all cases.
[0243] As expected, crystallization peak was displaced to lower
temperatures when increasing the weight of the sample or the
concentration of the solution.
[0244] However no significant difference was detected among the
different concentrations.
[0245] Detected differences are more linked to the internal
variability of samples.
[0246] A freezing end temperature of -45.degree. C. as well as a
freezing rate of 1.degree. C./min was chosen to ensure a full
crystalline state of the Vancomycin HCl during freezing.
[0247] Since the maximum allowable product temperature during
initial primary drying was -25.degree. C., the pressure during
primary drying was within 1/4 to 1/2 of the vapor pressure of ice
at -25.degree. C. Vapor of ice at -25.degree. C. is 630 .mu.bar.
The average of the thresholds, 230 .mu.bar, was selected as the
maximum allowed chamber pressure for primary drying.
[0248] Manufacturing Process
[0249] Water for injection was weighed out in a glass container on
calibrated balances.
[0250] Vancomycin HCl was added under stirring; the solution was
agitated until vancomycin was completely dissolved and the
dissolution time was recorded.
[0251] Then, water for injection was added until the required final
amount was reached.
[0252] On the final solution, pH and density were measured and
appearance was evaluated.
[0253] The solution was filtered through a 0.22 .mu.m PVDF
membrane.
[0254] The vials were washed with distilled water and dried in an
oven at 120.degree. C. for 2 h.
[0255] The filling was performed by mass and the in process
controls were carried out by weighing the filled vials every 20
vials.
[0256] After lyophilization the following analyses were performed
on the final product:
[0257] water content by Karl Fischer titration; appearance of the
cake, reconstitution time, appearance/clarity, pH after
reconstitution.
[0258] RP-HPLC was run to confirm processing did not influence
purity of vancomycin.
[0259] Twenty (20) ml of reconstituted drug product were passed
through the sterility testing membrane to confirm formulation
compatibility.
Example 2A
[0260] After evaluation of the DSC results, the following
lyophilization cycle nominal parameters were planned for use on the
100 mg/ml solution:
TABLE-US-00006 Step Time N.degree. Description Temperature
(.degree. C.) Pressure (hh:mm) 1 Load 20 Atmospheric NA 2 Product
freezing 20.fwdarw.-45 Atmospheric 01:05 3 Freeze soak time -45
Atmospheric 03:00 4 Evacuation -45 100 .mu.bar 00:01 5 Primary
drying -45.fwdarw. 10 100 .mu.bar 08:00 6 Primary drying 10 100
.mu.bar 14:00 7 Secondary drying 10 .fwdarw. 40 100 .mu.bar 00:30 8
Secondary drying 40 100 .mu.bar 09:00 9 Pre-aeration 0.95 bar NA 10
Stoppering 0.95 bar NA 11 Aeration Atmospheric NA Total length
35:36
[0261] The freezing soak and primary drying times were shortened
with respect to the set lyophilization program.
[0262] Actually, the product reached -45.degree. C. after 80
minutes of the freezing soak step. It was been kept at -45.degree.
C. one hour more and then the vacuum was pulled in the chamber to
start primary drying.
[0263] During step 6 (primary drying), all the product temperature
probes reached the temperature of the shelves (10.degree. C.) after
450 minutes.
[0264] The product was left at 10.degree. C. for 1 hour; afterwards
several pressure raise tests were performed to evaluate the
sublimation rate. The positive results of these tests allowed to
start heating to 40.degree. C. for secondary drying. Step 6 lasted
510 minutes instead of 840 minutes.
[0265] Total length of the cycle was 29 hours.
[0266] The cake had a cohesive structure that prevented loss of
friable material from the container during sublimation; lyophilised
product was not really elegant because of some cracks in the cake
(see the picture 1).
Example 2B
[0267] In this Example involving 100 mg/ml solution, 6R vials were
used. In this regard, twenty (20) mm neck vials enable a faster
sublimation than the 13 mm neck vials.
[0268] An intermediate step at 0.degree. C. during the primary
drying was inserted to have slower water vapor flow during
sublimation. In this way less cracks in the lyophilization cake
were observed.
[0269] The secondary drying temperature was reduced from 40.degree.
C. to 30.degree. C. according a client's request.
[0270] Final primary drying temperature was increased from
10.degree. C. to 15.degree. C. to try to maintain the total length
of the cycle to about 29 hours.
[0271] The nominal lyophilization parameters for this Example
were:
TABLE-US-00007 Step Time N.degree. Description Temperature
(.degree. C.) Pressure (hh:mm) 1 Load 20 Atmospheric NA 2 Product
freezing 20.fwdarw.-45 Atmospheric 01:05 3 Freeze soak time -45
Atmospheric 03:00 4 Evacuation -45 100 .mu.bar 00:01 5 Primary
drying -45.fwdarw. 0 100 .mu.bar 04:00 6 Primary drying 0 100
.mu.bar 02:00 7 Primary drying 0 .fwdarw. 15 100 .mu.bar 02:00 8
Primary drying 15 100 .mu.bar 10:00 9 Secondary drying 15 .fwdarw.
30 100 .mu.bar 00:15 10 Secondary drying 30 100 .mu.bar 09:00 11
Pre-aeration 0.95 bar NA 12 Stoppering 0.95 bar NA 13 Aeration
Atmospheric NA
[0272] The lower secondary drying temperature did not allow the
product to maintain a relatively low residual moisture. The overall
average value was 3.61 wt %, while the average moisture content of
previous batch was 1.71 wt %.
Example 2C
[0273] In this Example involving 100 mg/ml solution, the pressure
in the chamber was increased from 100 .mu.bar to 200 .mu.bar; a
higher pressure will favor the thermal exchanges at the gas/product
interface and the thermal conductivity from the shelf to the tray.
The bigger amount of heat transported to the product should result
in a rise of product temperature and consequently in a faster ice
sublimation.
[0274] Furthermore, after the evaluation of the lyophilization
printout, 4 hours were cut from the primary drying and added the
secondary drying step.
[0275] This Example involved the following nominal parameters:
TABLE-US-00008 Step Time N.degree. Description Temperature
(.degree. C.) Pressure (hh:mm) 1 Load 20 Atmospheric NA 2 Product
freezing 20.fwdarw.-45 Atmospheric 01:05 3 Freeze soak time -45
Atmospheric 03:00 4 Evacuation -45 200 .mu.bar 00:01 5 Primary
drying -45.fwdarw. 0 200 .mu.bar 04:00 6 Primary drying 0 200
.mu.bar 02:00 7 Primary drying 0 .fwdarw. 15 200 .mu.bar 02:00 8
Primary drying 15 200 .mu.bar 06:00 9 Secondary drying 15 .fwdarw.
30 200 .mu.bar 00:15 10 Secondary drying 30 200 .mu.bar 13:00 11
Pre-aeration 0.95 bar NA 12 Stoppering 0.95 bar NA 13 Aeration
Atmospheric NA Total length 31:21
[0276] Secondary drying was shortened from the programmed 780
minutes to 450 minutes. Actually, the product temperature matched
the shelf temperature very soon due to the better heat exchange by
drying at 200 .mu.bar.
[0277] The total length of the cycle was 24.5 hours.
[0278] Average moisture content was 1.82 wt %.
[0279] The lyophilization product still showed cracks in the
cake.
Example 2D
[0280] This Example involves a filling solution of 120 mg/ml to
allow doses of 120 mg, 240 mg, and 480 mg per vial.
[0281] All three fill volumes were lyophilized using the cycle for
the larger fill sample without paying attention to a possible over
drying of the lower fill volume samples.
[0282] The vancomycin 120 mg/mL filling solution was investigated
by performing a scansion with the differential calorimeter, and it
has been verified that the main thermal events were very close to
the ones detected on the 100 mg/mL filling solution.
[0283] This meant that the same lyophilization cycle conditions
were used for the 100 mg/mL could be applied to the 120 mg/mL.
[0284] New holding time studies were also performed on the 120
mg/mL concentration. The new cycle was tested on the 480 mg/vial
presentation that had the higher fill volume: 4 mL/vial.
[0285] The following nominal parameters were tested:
TABLE-US-00009 Step Time N.degree. Description Temperature
(.degree. C.) Pressure (hh:mm) 1 Load 20 Atmospheric NA 2 Product
freezing 20.fwdarw.-45 Atmospheric 01:05 3 Freeze soak time -45
Atmospheric 02:00 4 Evacuation -45 200 .mu.bar 00:01 5 Primary
drying -45.fwdarw. 0 200 .mu.bar 04:00 6 Primary drying 0 200
.mu.bar 02:00 7 Primary drying 0 .fwdarw. 15 200 .mu.bar 02:00 8
Primary drying 15 200 .mu.bar 24:00 9 Secondary drying 15 .fwdarw.
30 200 .mu.bar 00:15 10 Secondary drying 30 200 .mu.bar 15:00 11
Pre-aeration 0.95 bar NA 12 Stoppering 0.95 bar NA 13 Aeration
Atmospheric NA Total length 50:21
[0286] An overall average moisture content value of 0.97 wt % was
found by Karl Fisher titration.
Example 2E
[0287] Following the evaluation of the product temperature profile
versus the shelves temperature, the following run was cut three to
four hours in the primary drying step and four hours in the
secondary drying.
[0288] The 120 mg and the 240 mg units were placed in the
lyophilizer during the 480 mg cycle to check if over drying will
affect the chemical stability of the 120 mg and 240 mg vials.
[0289] The average residual moisture was 0.97 wt % for the 4 ml
fill, 1.23 wt % for the 2 ml fill, and 1.34 wt % for the 1 ml.
[0290] The lyophilization cycle had a total length of nearly 42
hours.
TABLE-US-00010 Step Time N.degree. Description Temperature
(.degree. C.) Pressure (hh:mm) 1 Load 20 Atmospheric NA 2 Product
freezing 20.fwdarw.-45 Atmospheric 01:05 3 Freeze soak time -45
Atmospheric 02:00 4 Evacuation -45 200 .mu.bar 00:01 5 Primary
drying -45.fwdarw. 0 200 .mu.bar 04:00 6 Primary drying 0 200
.mu.bar 02:00 7 Primary drying 0.fwdarw. 15 200 .mu.bar 02:00 8
Primary drying 15 200 .mu.bar 20:00 9 Secondary drying 15 .fwdarw.
30 200 .mu.bar 00:15 10 Secondary drying 30 200 .mu.bar 11:00 11
Pre-aeration 0.95 bar NA 12 Stoppering 0.95 bar NA 13 Aeration
Atmospheric NA Total length 42:21
[0291] All three presentations had cake with a very cohesive
structure even if some cracks were present.
Analytical Results
TABLE-US-00011 [0292] In Process Controls Results Process step
Analytical Test 2A 2B 2C 2D 2E Formulated pH 3.90 3.86 3.83 3.69
3.72 Bulk Density (g/mL) 1.027 1.028 1.030 1.0394 1.0389
solution
TABLE-US-00012 Tests on Freeze-dried drug product Results Process
step Analytical Test 2A 2B 2C 2D 2E Final Water content 1.71 3.61
1.82 0.97 See Lyophilizate by KF [% w/w] below Visual aspect of
Conform Conform Conform Conform Conform the cake Reconstitution
~30'' ~30'' ~30'' ~5'' See below time Appearance of Conform Conform
Conform Conform Conform reconstituted solution (water, 50 mg/ml) pH
3.54 3.53 3.54 3.30 See below % Vancomycin 93.0 92.9 92.9 92.0 See
B HCl [HPLC] below % impurities 7.0 7.2 6.9 8.0 See below
TABLE-US-00013 Moisture Content (K.F.) [wt %] Results 2E Sample 2A
2B 2C 2D 120 mg 240 mg 480 mg Front sample 2.06 3.54 1.92 1.02 1.41
1.26 0.96 Middle sample 1.40 3.12 1.80 1.01 1.27 1.15 0.97 Back
sample 1.68 4.17 1.74 0.92 1.27 1.27 0.97 Average 1.71 3.61 1.82
0.97 1.34 1.23 0.97
TABLE-US-00014 pH Results 2E Sample 2A 2B 2C 2D 120 mg 240 mg 480
mg Front sample 3.49 3.54 3.56 3.32 3.36 3.39 3.31 Middle sample
3.57 3.52 3.52 3.29 3.36 3.38 3.33 Back sample 3.58 3.52 3.54 3.30
3.37 3.39 3.31 Average 3.54 3.53 3.54 3.30 3.36 3.39 3.32
TABLE-US-00015 % Content Vancomycin B Hydrochloride (% VMB) Results
Reference Std. Sample Vancomycin 2A 2B 2C 2E 2D Front sample 93.7
92.9 92.9 92.9 93.7 120 mg 91.3 240 mg 91.6 480 mg 92.5 Middle
sample 93.0 93.0 93.0 92.1 120 mg 91.8 240 mg 91.7 480 mg 92.1 Back
sample 93.1 93.0 93.0 91.9 120 mg 92.1 240 mg 91.2 480 mg 91.8
Average 93.0 92.9 92.9 92.0 120 mg 91.7 240 mg 91.5 480 mg 92.1
Standard Dev. 0.104 0.0327 0.0327 0.108 120 mg 0.415 240 mg 0.261
480 mg 0.343 % RSD 0.112 0.0352 0.0352 0.117 120 mg 0.453 240 mg
0.285 480 mg 0.372
TABLE-US-00016 Related substances (% impurities) Results Reference
Std. Sample Vancomycin 2A 2B 2C 2D 2E Front sample 6.3 7.1 7.2 6.9
6.3 120 mg 8.8 240 mg 8.4 480 mg 7.5 Middle 7.1 7.3 6.9 7.9 120 mg
8.2 sample 240 mg 8.3 480 mg 7.9 Back 6.9 7.1 7.0 8.1 120 mg 7.9
sample 240 mg 8.8 480 mg 8.3 Average 7.0 7.2 6.9 8.0 120 mg 8.3 240
mg 8.5 480 mg 7.9 Standard 0.12 0.09 0.013 0.11 120 mg 0.42 Dev.
240 mg 0.26 480 mg 0.42 % RSD 1.7 1.2 0.23 1.3 120 mg 5.0 240 mg
3.1 480 mg 5.3
[0293] Reconstitution Time
[0294] Reconstitution time measurement was carried out adding:
[0295] 1.0 mL of WFI to the 120 mg/vial strength [0296] 2.0 mL of
WFI to the 240 mg/vial strength [0297] 4.0 mL of WFI to the 480
mg/vial strength
[0298] The observed reconstitution time on the product of Example
2E was quite short relative to all the tested vials; about 10
seconds were needed to completely reconstitute the 120 mg
freeze-dried drug product; 10 to 15 seconds were needed to
completely reconstitute the 240 mg/vial presentation, while about
20 seconds were needed to completely reconstitute the 480 mg
units.
[0299] The reconstituted solution had a clear, light pinkish
appearance and was particle free.
[0300] Compatibility with Sterility Testing Membrane
[0301] 20 mL of reconstituted drug product were passed through the
sterility testing membrane to confirm the formulation
compatibility.
[0302] The solution passed through the filter membrane, and 17 ml
of the 20 ml were collected below the membrane.
Example 3
Summary
[0303] This Example involves a freeze-drying cycle for a 600 mg of
Vancomycin HCl/vial strength.
[0304] Materials/Equipment
[0305] Materials [0306] Vancomycin hydrochloride, USP,
Alpharma--Denmark [0307] ISO 6R clear type I glass vials, Nuova
Ompi--Italy [0308] 20 mm freeze-drying stoppers, West
Pharmaceutical Service--USA [0309] 20 mm flip-off caps, Capsulit
S.p.A.--Italy [0310] 13 mm freeze-drying stoppers, West
Pharmaceutical Service--USA [0311] 13 mm flip-off caps, West
Pharmaceutical Service--USA
[0312] Equipment [0313] Glassware for Vancomycin solution before
and after filtration (bottles). [0314] Pressure vessel,
Sartorius--Germany [0315] Balance to check the filling weight (10
mg sensitivity), Sartorius--Germany [0316] Digital pH meter,
Mettler Toledo--Switzerland [0317] Karl Fischer automatic titrator
DL38, Mettler Toledo--Switzerland [0318] 0.22 .mu.m sterilizing
PVDF filter, Pall [0319] Semiautomatic filling machine, Flexicon
PF6--Denmark [0320] Isolator, E.Co.Tec--Italy [0321] Lyophilizer,
BOC Edwards Lyoflex 04 (or Minifast 8000) with the following
characteristics: 0.4 m.sup.2 (or 0.8 m.sup.2 shelf surface, shelf
temperature range was -50.degree. C. to +50.degree. C., PT 100
temperature probes, Pirani gauge for vacuum monitoring, coil
condenser with ice capacity of 8 kg, condenser coil inlet,
temperature arrives to -60.degree. C., stainless steel trays with a
thickness of about 2 mm [0322] Semiautomatic crimping machine,
Flexseal--Denmark
[0323] Formulation
[0324] Vancomycin hydrochloride was dissolved in water for
injection to form a 120 mg/ml formulation, as shown below.
TABLE-US-00017 Quantity Ingredients Amount/ml Amount/Unit
Vancomycin HCl 120.00 600.00 mg Water for injection to 1.00 ml to
5.00 ml
[0325] Manufacturing Process
[0326] Water for injection was weighed out in a glass container on
calibrated balances.
[0327] Vancomycin HCl was added under stirring; the solution was
agitated until vancomycin was completely dissolved and the
dissolution time was recorded.
[0328] Then, water for injection was added until the required final
amount was reached.
[0329] On the final solution, pH and density were measured and
appearance was evaluated.
[0330] The solution was filtered through a 0.22 .mu.m PVDF
membrane.
[0331] The vials were washed with distilled water and dried in an
oven at 120.degree. C. for 2 h.
[0332] The filling was performed by mass and the in process
controls were carried out by weighing the filled vials every 20
vials.
[0333] After lyophilization the following analyses were performed
on the final product:
[0334] water content by Karl Fischer titration;
[0335] appearance of the cake,
[0336] reconstitution time,
[0337] appearance/clarity,
[0338] pH after reconstitution.
[0339] RP-HPLC was run to confirm processing didn't influence
purity of Vancomycin.
[0340] Lyophilization Cycle
[0341] The product was freeze-dried according the following nominal
lyophilization cycle parameters:
TABLE-US-00018 Step Time N.degree. Description Temperature
(.degree. C.) Pressure (hh:mm) 1 Load 20 Atmospheric NA 2 Product
freezing 20.fwdarw.-45 Atmospheric 01:05 3 Freeze soak time -45
Atmospheric 02:00 4 Evacuation -45 200 .mu.bar 00:01 5 Primary
drying -45.fwdarw. 0 200 .mu.bar 04:00 6 Primary drying 0 200
.mu.bar 02:00 Primary drying 0.fwdarw. 15 200 .mu.bar 02:00 Primary
drying 15 200 .mu.bar 24:00 7 Secondary drying 15 .fwdarw. 30 200
.mu.bar 00:15 8 Secondary drying 30 200 .mu.bar 15:00 9 Preaeration
0.95 bar NA 10 Stoppering 0.95 bar NA 11 Aeration Atmospheric NA
Total length (without stoppering) 50:21
[0342] Results
[0343] An overall average moisture content value of 1.04 wt % was
found by Karl Fisher titration.
[0344] Cakes had a very cohesive structure even if some cracks were
present.
[0345] Analytical Results
TABLE-US-00019 In Process Controls Process step Analytical Test
Results Formulated pH 3.69 Bulk Density (g/mL) 1.0384 solution
Concentration (UV) 116.88 mg/mL
TABLE-US-00020 Tests on Freeze-dried Drug Product Process step
Analytical Test Results Final Water content by K F 1.04% w/w
Lyophilizate Visual aspect of the cake Whitish solid compact mass
Reconstitution time 30 seconds Appearance of Clear, colourless
solution reconstituted solution (water, 50 mg/ml) pH 3.44 %
Vancomycin B by 93.3% RP-HPLC
TABLE-US-00021 Moisture content (K. F.) Sample Moisture Sample 1
(back) 1.07% Sample 2 (middle) 1.02% Sample 3 (front) 1.02% Overall
average 1.04%
TABLE-US-00022 HPLC Assay (% Vancomycin B) Sample Vancomycin B
Sample 1 (back) 93.3% Sample 2 (middle) 93.3% Sample 3 (front)
93.3% Overall average 93.3%
TABLE-US-00023 pH Sample pH Sample 1 (back) 3.45 Sample 2 (middle)
3.44 Sample 3 (front) 3.44 Overall average 3.44
[0346] Reconstitution Time
[0347] About 30 seconds were needed to completely reconstitute the
freeze-dried drug product with 5.0 mL of WFI. The reconstituted
solution had a clear, colorless appearance and was particle
free.
Example 4
Summary
[0348] Nebulization characteristics of gentamicin and vancomycin
solutions were evaluated as a function of solution strength,
nebulizer fill volume, and saline concentration. Key aerosol
attributes measured were emitted dose and particle size
distributions. All experiments were performed using Aerotech II jet
nebulizers operated continuously at 8 LPM. For gentamicin solutions
in WFI, the range of solution strengths varied from 40 to 120
mg/ml, and fill volumes ranged from 2 to 4 ml. The resulting
aerosol dose emitted over 30 minutes of nebulization was found to
vary from 40 mg to over 300 mg, with the dose increasing
proportionally with increasing fill volume and solution strength.
Emitted dose measurements for vancomycin were performed for
solutions in normal saline, in 0.45% saline, and in water for
injection. The range of solution concentrations tested ranged from
60 mg/ml to 140 mg/ml. The cumulative aerosol dose emitted for a 30
minute nebulization period varied from about 50 mg to over 300 mg,
with the dose increasing proportionally with solution strength and
fill mass.
[0349] Particle size distributions were measured for the above drug
solutions using a laser diffraction spectrometer. The median
particle size for all solutions tested was in the range 2-3 .mu.m,
well within the respirable size range. Particle size distributions
for these antibiotic drugs were found to be relatively insensitive
to solution strength and fill volume. Follow-on measurements with
drug and normal saline solutions indicated that the size
distribution of nebulized antibiotics were comparable to that for
the normal saline solution.
[0350] Combined together, the above results indicate that a broad
range of aerosol doses in the respirable range may be achieved for
nebulized vancomycin and gentamicin by suitably selecting nebulizer
fill volume and solution strengths.
[0351] Objectives
[0352] To determine the amount of drug aerosol emitted during the
nebulization of gentamicin and vancomycin solutions, as a function
of nebulizer fill volume and solution strength.
[0353] To determine the size distribution of aerosols produced
during the nebulization of gentamicin, vancomycin, and saline
solutions as a function of nebulizer fill volume and solution
strength.
[0354] Introduction
[0355] This Example involves assessing nebulization characteristics
such as the emitted dose and droplet size distribution for
antibiotic drug solutions of different strengths and at different
nebulizer fill volumes. The emitted dose information is useful in
selecting solution strengths and fill masses to deliver a chosen
target dose. The particle size information is useful in determining
whether the aerodynamic size of the aerosol produced is in the
range required for effective lung deposition (1-5 .mu.m). Results
for a placebo solution (i.e., normal saline) are also reported for
comparison. All of the experiments were performed using an Aerotech
II jet nebulizer operated continuously at a nominal flow rate of 8
LPM. Aerosol emitted dose was estimated by using filters to collect
the aerosol output generated by the nebulizer, and assaying the
amount of drug deposited. Particle size distributions of the
generated aerosol were measured using a Sympatec laser diffraction
spectrometer.
[0356] Study Design
[0357] Characterization of Emitted Dose
[0358] For the case of gentamicin solution in water, a full
factorial experiment was performed to characterize emitted mass of
aerosol as a function of two factors, i.e. nebulizer fill volume
and fill mass. The range of solution strengths and fill volume was
chosen to provide a broad range of target doses achievable with a
nebulization time of 30 minutes.
[0359] The test matrix for this experiment is presented in Table 1.
Gentamicin solution strength (based on mass of drug) was varied
from 40 mg/ml to 120 mg/ml, while the nebulizer fill volume was
varied from 2 to 4 ml. Each of the 9 treatment combination was
repeated twice, for a total of 18 runs. The gentamicin solutions
were prepared in water for injection (WFI), and were preservative
free.
[0360] For the case of vancomycin, the emitted mass of aerosol was
characterized for following three cases: [0361] Vancomycin in
normal saline, solution strength of 60 mg/ml, nebulizer fill volume
ranging from 2-4 ml. [0362] Vancomycin in 0.45% saline, solution
strength ranging from 60-90 mg/ml, nebulizer fill volume ranging
from 2-4 ml. [0363] Vancomycin in WFI, solution strength ranging
from 60-140 mg/ml, nebulizer fill volume ranging from 2-4 ml.
[0364] In the case of vancomycin, addition of salt to the
formulation allows for tuning of solution properties such as
osmolality. Test matrices for the above three experiments are
presented in Tables 2-4.
TABLE-US-00024 TABLE 1 Test Matrix for Gentamicin Solution in WFI
Fill Volume Solution Strength Pattern [ml] [mg/mL] 13 2 120 31 4 40
22 3 80 12 2 80 11 2 40 21 3 40 21 3 40 13 2 120 23 3 120 31 4 40
33 4 120 11 2 40 22 3 80 12 2 80 23 3 120 33 4 120 32 4 80 32 4
80
TABLE-US-00025 TABLE 2 Test Matrix for Vancomycin Solution (60
mg/ml) in Normal Saline Vancomycin at 60 mg/ml (in normal saline)
Fill volume 2 ml Fill volume 3 ml Fill volume 2 ml Fill volume 4 ml
Fill volume 4 ml Fill volume 3 ml Fill volume 3 ml Fill volume 4 ml
Fill volume 2 ml
[0365] The responses measured for all of the above experiments
included:
[0366] (i) the mass of drug delivered in 15 mins
[0367] (ii) the cumulative mass of drug delivered in 30 mins,
and
[0368] (iii) the mass of drug remaining in the nebulizer after 30
mins of operation.
TABLE-US-00026 TABLE 3 Test Matrix for Vancomycin Solution in 0.45%
Saline Fill Volume Solution Strength Pattern [ml] [mg/L] 11 2 60 13
2 90 13 2 90 11 2 60 32 4 75 23 3 90 12 2 75 21 3 60 23 3 90 32 4
75 21 3 60 33 4 90 12 2 75 33 4 90 22 3 75 31 4 60 31 4 60 22 3
75
TABLE-US-00027 TABLE 4 Test Matrix for Vancomycin Solution in WFI
Fill Volume Solution Strength Pattern [ml] [mg/ml] 11 2 60 13 2 140
13 2 140 11 2 60 32 4 100 23 3 140 12 2 100 21 3 60 23 3 140 32 4
100 21 3 60 33 4 140 12 2 100 33 4 140 22 3 100 31 4 60 31 4 60 22
3 100
[0369] Characterization of Particle Size Distribution
[0370] For the case of gentamicin solution in water, a full
factorial experiment experiment was performed to characterize the
particle size distribution of aerosol as a function of two factors,
i.e. nebulizer fill volume and fill mass. The test matrix for this
experiment is presented in Table 5. Gentamicin solution strength
(based on mass of drug) was varied from 40 mg/ml to 120 mg/ml,
while the nebulizer fill volume was varied from 2 to 4 ml. The 9
treatment combinations were run in a random order. A fresh
nebulizer was used for each run. The nebulizers in this experiment
were prequalified using a flow rate test to minimize variability in
the test results.
TABLE-US-00028 TABLE 5 Test Matrix for Gentamicin Solution in WFI
Fill Volume Solution Strength Run Pattern [mL] [mg/mL] 1 31 4 40 2
32 4 80 3 21 3 40 4 23 3 120 5 22 3 80 6 13 2 120 7 11 2 40 8 33 4
120 9 12 2 80
[0371] For the case of vancomycin, the emitted mass of aerosol was
characterized for following three cases: [0372] Vancomycin in
normal saline, solution strength of 60 mg/ml, nebulizer fill volume
ranging from 2-4 ml. [0373] Vancomycin in 0.45% saline, solution
strength ranging from 60-90 mg/ml, nebulizer fill volume ranging
from 2-4 ml. [0374] Vancomycin in WFI, solution strength ranging
from 60-140 mg/ml, nebulizer fill volume ranging from 2-4 ml.
[0375] The test matrices for the above three experiments are
presented in Tables 6-8. A fresh nebulizer was used for each run.
The nebulizers in these experiments were pre-screened using a flow
rate test to minimize variability in the test results.
TABLE-US-00029 TABLE 6 Test Matrix for Vancomycin Solution (60
mg/ml) in Normal Saline Fill Volume Solution Strength Run Pattern
[mL] [mg/mL] 1 1 2 60 2 3 4 60 3 2 3 60
TABLE-US-00030 TABLE 7 Test Matrix for Vancomycin Solution in 0.45%
Saline Fill Volume Solution Strength Run Pattern [mL] [mg/mL] 1 12
2 75 2 31 4 60 3 22 3 75 4 33 4 90 5 13 2 90 6 11 2 60 7 32 4 75 8
21 3 60 9 23 3 90
TABLE-US-00031 TABLE 8 Test Matrix for Vancomycin Solution in WFI
Fill Volume Solution Strength Run Pattern [mL] [mg/mL] 1 33 4 140 2
22 3 100 3 13 2 140 4 12 2 100 5 11 2 60 6 32 4 100 7 21 3 60 8 23
3 140 9 31 4 60
[0376] A follow on experiment was performed to characterize
particle size distributions of aerosols generated using vancomycin
and gentamicin solutions in water at a fixed solution strength of
120 mg/ml, and a fixed fill volume of 5 ml. Particle size
distributions of drug aerosol were compared against those obtained
by nebulizing normal saline solution at a fill volume of 5 ml. The
test matrix for this follow on experiment is presented in Table 9.
Each treatment was repeated 3 times.
TABLE-US-00032 TABLE 9 Test Matrix for Evaluation of Drug and
Placebo Solutions Fill Volume Run Drug [mL] 1 Normal Saline 5 2
Vancomycin 5 3 Gentamicin 5 4 Gentamicin 5 5 Normal Saline 5 6
Gentamicin 5 7 Vancomycin 5 8 Vancomycin 5 9 Normal Saline 5
[0377] Equipment and Materials
[0378] Equipment [0379] Sympatec HELOS Magic BFS laser diffraction
spectrometer, Ser. No. 085 [0380] Mass flow meter (TSI 4000 series)
[0381] Rotameter [0382] Volumetric flow meter, Dry Cal [0383]
Pressure regulator [0384] Flow regulating valve [0385] Flow
shut-off valve [0386] Pipet
[0387] Materials [0388] Aerotech II Nebulizer [0389] Tee connector
and mouthpiece from Hudson RCI MicroMist Nebulizer (Cat No. 1882)
[0390] Inspiratory filter (PARI electret filter) [0391] Filter
holder [0392] One way valve [0393] 50 ml centrifuge tubes [0394]
HPLC water [0395] HPLC water dispenser [0396] Vancomycin HCl [0397]
Gentamycin Sulfate
[0398] Procedure
[0399] Characterization of Emitted Dose
[0400] The nebulizer was connected to a standard "T" piece coupled
to a filter holder on one end, and a flow inlet channel provided
with a one-way valve on the other end. The filter holder supported
a PARI electret filter used to collect the aerosol dose emitted by
the nebulizer.
[0401] The nebulizer was operated using clean, dry compressed air
from a source regulated to a pressure of about 50 psig. The flow
rate of air through the nebulizer was controlled using a rotameter
and set to a nominal flow rate 8 LPM. The drug laden air from the
nebulizer passed through the collection filter into an exhaust line
provided with a backup filter and a flow regulating valve, and
connected to a vacuum source. The flow regulating valve was set so
that the vacuum suction flow was slightly higher than the nebulizer
output flow. A small amount of clean make up air was allowed to
enter through the one way valve to make up for the flow deficit.
This arrangement enabled efficient collection of the nebulizer drug
output by the filter. The emitted dose experiments were performed
with the nebulizer operating continuously at 8 LPM for a total
nebulization time of 30 minutes. The filter/filter holder were
replaced with a fresh filter/filter holder at the 15 minute point,
so that the accumulated drug output at 15 minutes and 30 minutes
could be evaluated. The filter samples were placed in centrifuge
tubes and rinsed with a pre-determined amount of HPLC water
(ranging from 30-40 ml). Residual drug from each filter holder was
also rinsed into the corresponding centrifuge tube using some of
the filter rinsate. The residual drug from the nebulizer was also
rinsed into a 50 ml centrifuge tube using a pre-determined amount
of HPLC water (ranging from 30-40 ml). The drug content of the
filter and nebulizer samples were assessed by drug specific HPLC
assays. Note that the measurement of filter and nebulizer samples
permit a full mass balance to be performed for each run.
[0402] Characterization of Particle Size Distribution
[0403] Droplet size distributions for aerosolized drug and placebo
solutions were measured using the Sympatec HELOS laser diffraction
spectrometer. In preparation for a run, the nebulizer was connected
to the compressed air line, the flow turned on and the pressure
regulator set to a driving pressure to generate a flow rate of 8
LPM through the nebulizer. The flow was then turned off by closing
the flow shut-off valve. Next, the nebulizer was connected to a "T"
piece with one port plugged, and the other port coupled to a
mouthpiece. The nebulizer was then filled with drug solution, and
mounted so that nebulizer mouthpiece was aligned parallel to the
nozzle of the Rodos dry powder disperser apparatus already
installed in the spectrometer. The laser diffraction system was
setup to automatically trigger when it sensed the presence of the
aerosol generated by the nebulizer. Measurements were initiated by
opening the shut-off valve to pressurize the nebulizer and generate
the aerosol. A total of 6 particle size distribution scans were
taken for each nebulizer run, and then averaged to provide
representative size distribution results.
Results and Discussion
[0404] Characterization of Emitted Dose
[0405] Summarized dose delivery results for the case of gentamicin
solutions are presented in FIGS. 3-5. FIG. 3 is a bar graph showing
the total drug recovered from the nebulizer and as a function of
nebulizer fill volume and solution strength. Each recovery value is
the average of two replicate runs (run order listed in Table1). The
drug recovery was very consistent across solution strengths and
fill volumes, varying in the range 97.1%-101.2% of fill mass,
indicating that a full mass balance was achieved from these
measurements.
[0406] FIGS. 4a and 4b present the cumulative emitted dose of
gentamicin, respectively at the 15 min and 30 min time points, as a
function of fill volume and solution strength. Again, each value
reported is the average of two replicate runs. The delivered dose
was observed to increase with an increase in both fill volume and
solution strength, consistent with expectation. A comparison of
these two figures shows that the collected dose at 15 minutes was
comparable to that at 30 minutes for 2 and 3 ml fill volumes,
indicating that the dose emission at these fill volumes occurred
within 15 minutes. For the 4 ml fill volume, the collected dose at
30 mins was only slightly larger than the value at 15 minutes,
indicating that nebulization was largely completed within the 15
minute period. From this it can be concluded that fill volumes of
up to 4 ml of gentamicin solution of strengths up to 120 mg/ml can
be effectively nebulized within a duration of 30 minutes. FIG. 4b
also indicates that a gentamicin aerosol doses spanning a factor of
up to 7 can be delivered from the nebulizer by suitably tuning the
solution strength and fill volume within the ranges tested.
[0407] FIG. 5 presents the gentamicin dose retained by the
nebulizer at the end of 30 minutes, as a function of solution
strength and fill volume. The values reported are averages of two
replicate runs. The retained dose was found to increase with
increasing solution strength and fill volume, with a steeper
increase observed with increasing solution strength.
[0408] Similar trends in emitted dose as a function of solution
strength and fill volume were obtained for the case of vancomycin.
Illustrative emitted dose measurements for vancomycin are presented
in FIGS. 6-8.
[0409] For the case of 60 mg/ml solution in normal saline (see
Table 2), FIG. 6 plots the distribution of vancomycin drug after 30
minutes of nebulization as a function of fill volume. The plot
shows the dose retained in the nebulizer and that collected at the
15 minute (filter 1) and 30 minute (filter 2) timepoint. The
reported values are averages calculated for 3 replicate runs. As
with the case of gentamicin, dose emission was found to be largely
completed within 15 minutes, and the accumulated dose (i.e. filter
1+filter 2) at the end of 30 minutes was found to increase with
fill volume.
[0410] For the case of vancomycin solutions in 0.45% saline (see
test matrix in Table 3), FIG. 7 plots the cumulative emitted dose
after 30 minutes of nebulization as a function of solution strength
and fill volume. The delivered dose was observed to increase with
increasing fill volume and solution strength, as expected. FIG. 8
plots similar results for the case of vancomycin solutions in WFI,
obtained for the test matrix presented in Table 4.
[0411] It is clear from FIGS. 6-8 that aerosol doses of vancomycin
spanning a six fold range can be obtained from the nebulizer by
suitably tuning the fill volume and solution strength within the
ranges tested.
[0412] Characterization of Particle Size Distribution
[0413] Representative laser diffraction particle size measurements
for the case of gentamicin solutions (test matrix of Table 5) are
summarized in FIGS. 9 and 10. FIG. 9 plots the volume median
diameter for aerosolized gentamicin as a function of fill volume
and solution strength (test matrix in Table 5). Each reported value
was obtained by averaging 6 replicate laser diffraction
measurements for each nebulization run. The measured median
particle size for all of the gentamicin solutions varied slightly
in the 2-3 .mu.m range, and appeared to be relatively insensitive
to fill volume or solution strength. In all cases, the median
particle diameter was well within the "respirable size range"
considered to be suitable for pulomary drug delivery (1-5 .mu.m).
FIG. 10 plots the cumulative volume weighted particle size
distributions for gentamicin aerosol for all of the solution
strengths and fill volumes tested. The size distributions obtained
for these solutions were observed to vary within a narrow range
over the fill volumes and solution strengths tested. FIG. 10 also
provides a measure of the spread of the aerosol size distribution,
and it was observed that a major fraction of the aerosol was within
the respirable size range.
[0414] Representative particle sizing measurements for vancomycin
solutions in WFI (test matrix in Table 8) are presented in FIGS. 11
and 12, and are roughly comparable to that obtained for gentamicin
solutions.
[0415] FIG. 11 indicates that the volume weighted median sizes for
these vancomycin solutions were largely within the range of 2-3
.mu.m, also well within the respirable range. The spreads of the
aerosol size distribution, shown in FIG. 12, were similar to that
obtained for nebulized gentamicin.
[0416] FIGS. 13 and 14 are plots of volume median diameter for the
case of vancomycin solutions in normal saline (test matrix in Table
6), and 0.45% saline (test matrix in Table 7) respectively,
obtained at different solution strengths and fill volumes. The size
distributions were found to be comparable to that obtained for the
vancomycin solutions in water. In general, the size distributions
of vancomycin solutions were largely insensitive to fill volume,
solution strength, and saline concentration.
[0417] Finally, results from the follow-on particle sizing study
with the test matrix listed in Table 9 are presented in FIG. 15.
This figure plots volume median diameters for solutions of
vancomycin (120 mg/ml), gentamicin (120 mg/ml) and normal saline,
all obtained for nebulizer fill volumes of 5 ml. For each solution,
results from three nebulizer runs are provided. It is seen from
this plot that the median particle size for all three solutions
were comparable and were in the 2-3 .mu.m range, well within the
respirable size range.
[0418] Conclusions
[0419] The emitted dose of nebulized gentamicin and vancomycin was
measured as a function of solution strength, fill volume, and
saline concentration. All experiments were performed using Aerotech
II jet nebulizers operated continuously at 8 LPM. For gentamicin
solutions in WFI, the range of solution strengths varied from 40 to
120 mg/ml, and fill volumes ranged from 2 to 4 ml. The resulting
aerosol dose emitted over 30 minutes of nebulization was found to
vary from 40 mg to over 300 mg, with the dose increasing with
increasing fill volume and solution strength. Emitted dose
measurements for vancomycin were performed for solutions in normal
saline, in 0.45% saline, and in water for injection. The range of
solutions tested ranged from 60 mg/ml to 140 mg/ml. The cumulative
aerosol dose emitted over a 30 minute nebulization period varied
from about 50 mg to over 300 mg, with the dose increasing with
solution strength and fill mass.
[0420] Particle size distributions were measured for the above drug
solutions using a laser diffraction spectrometer. The median
particle size for all solutions tested was in the range 2-3 .mu.m,
well within the respirable size range. Particle size distributions
for these antibiotic drugs were found to be relatively insensitive
to solution strength and fill volume. Follow-on measurements with
drug and normal saline solutions indicated that the size
distribution of nebulized antibiotics were comparable to that for
the normal saline solution.
[0421] Combined together, the above results demonstrate that a
broad range of aerosol doses in the respirable range may be
achieved for nebulized vancomycin and gentamicin by suitably
selecting nebulizer fill volume and solution strengths.
Example 5
[0422] This Example involved evaluating the potential toxicity and
recovery resulting from a 14-consecutive day, nose-only inhalation
administration of vancomycin hydrochloride (vancomycin) to CD
rats.
[0423] Within 2 hours prior to usage, a vancomycin nebulizer
solution having a concentration of 120 mg/ml (based on vancomycin
potency of bulk material) was formed by dissolving vancomycin
hydrochloride (available from Alpharma, Copenhagen, Denmark) in
sterile water for injection USP (available from B. Braun Medical
Inc., Bethlehem, Pa.). The solution was used to generate
aerosolized vancomycin for all vancomycin exposure groups.
[0424] Nose-only exposures were conducted in a "flow-past"
cylindrical inhalation chamber placed inside a steel-framed
Plexiglas secondary containment box. The chamber contained 48
animal ports, each compatible with a single nose-only exposure
tube, aerosol concentration sampling device (e.g., filter), or
oxygen monitor.
[0425] The total air flow through the exposure system was balanced
to achieve individual animal port flows of .about.500 mL/min (port
flow approximated based on total chamber flow). Measured flows
included sample flow rate, nebulizer flow rate, dilution flow rate
(chamber make-up air), and chamber exhaust flow. The exposure
chamber had a slightly higher exhaust flow rate than inlet flow
rate.
[0426] Vancomycin solution was aerosolized with two Aerotech II
nebulizers operated at 20 psi driving pressure. The target aerosol
Vancomycin concentration for all exposure levels was .about.1.0
mg/L.
[0427] Aerosolized vancomycin was administered to 3 groups of male
and female CD rats (available from Charles River Laboratories,
Kingston, N.Y.) for durations of 30 min (Low), 90 min (Mid), and
180 min (High). A control group was exposed for 180 min to aerosols
generated from a normal saline solution. Groups of rats from the
Control and High level 14-day exposures were also studied following
a 14-day recovery period. Endpoints included clinical observations,
body weights, clinical pathology (hematology, clinical chemistry),
urinalyses, organ weights, and histopathology.
[0428] Vancomycin aerosol concentrations were 1.23.+-.0.16,
1.25.+-.0.12, and 1.23.+-.0.08 mg/L for the Low, Mid, and High
exposure levels, respectively. Mean particle size was determined to
be in the inhalable size range for rodents (2.0-2.6 .mu.m mass
median aerodynamic diameter). Mean total inhaled doses were
estimated as 23, 71 and 139 mg/kg, and mean doses deposited in lung
were estimated as 3, 9, and 17 mg/kg for the Low, Mid, and High
exposure levels, respectively.
[0429] The vancomycin exposures were well-tolerated by all groups
of rats. All rats survived to scheduled necropsy, and there were no
vancomycin related effects noted on clinical observations. There
were also no vancomycin treatment related effects on body
weight.
[0430] The only organ weights to show consistent vancomycin related
effects were lungs. Lung weights were statistically significantly
increased by an average approximately 8, 20, 19% of control for the
Low, Mid and High exposure levels respectively.
[0431] Exposure related histopathologic findings were limited to
the respiratory tract. Observations included minimal to mild nasal
mucous cell hyperplasia and hypertrophy, minimal to mild pulmonary
interstitial inflammation and alveolar macrophage hyperplasia with
an apparent dose-response effect, lymphoid hyperplasia of the
tracheobronchial and mediastinal lymph nodes, and slight laryngeal
inflammation. There was substantial diminution of these findings
after 14 days of recovery with pulmonary interstitial inflammation,
alveolar macrophage hyperplasia, and nasal mucus cell hyperplasia
persisting in the high dose group, but at a lesser severity overall
than seen at the end of exposure. A threshold of response was not
established although the effects in the low dose group were
generally minimal.
[0432] Clinical pathology findings were generally unremarkable. The
only vancomycin related effect on hematology was a statistically
significant increase in neutrophils at the Mid and High exposure
levels. The only vancomycin related effect on clinical chemistry
was a mild but statistically significant increase in aspartate
aminotransferase (AST) values (.about.28-46%) at the Mid and High
exposure levels. Neutrophil changes were diminished after the
recovery period resolved. AST observations resolved after the
recovery period. Both findings likely resulted from the minimal to
mild pulmonary inflammation manifested in the histopathology
findings. No vancomycin related changes were seen after examination
of serum indicators of kidney function or urinalysis.
[0433] To conclude, the findings indicate that exposure to
vancomycin at the Mid level and High level exposures,
predominantly, caused an irritant reaction in the respiratory tract
manifested by minimal to moderate mucous cell changes in the nose
and minimal to mild inflammation and macrophage hyperplasia in the
lungs. Corresponding changes in neutrophil and AST values likely
resulted from the pulmonary inflammatory findings. Recovery from
these effects was evident, but not entirely resolved after the
14-day observation period. A no observed effect level was not
established.
Example 6
[0434] This Example involved evaluating the potential toxicity and
recovery resulting from a 14-consecutive day, face mask inhalation
administration of vancomycin hydrochloride to beagle dogs.
[0435] Within 2 hours prior to usage, a vancomycin nebulizer
solution having a concentration of 120 mg/ml was formed by
dissolving vancomycin hydrochloride (available from Alpharma,
Copenhagen, Denmark) in sterile water for injection USP (available
from B. Braun Medical Inc., Bethlehem, Pa.). The solution was used
to generate aerosolized vancomycin for all vancomycin exposure
groups.
[0436] The exposure system consisted of a single, cylindrical,
plexiglass inhalation chamber (volume of .about.23.7 L, 14.61-cm
radius, 35.56-cm height). The chamber was supplied with two
Aerotech II nebulizers operated at .about.40 psi. Nebulized test
article and nebulizer air supply was diluted with .about.10 L/min
HEPA-filtered dilution air. The flow through the system was
.about.36 L/min.
[0437] The aerosolized vancomycin was administered via a face mask
to 3 groups of male and female beagle dogs for durations of 15 min
(Low), 30 min (Mid), and 60 min (High). A control group was exposed
for 60 min to aerosols generated from normal saline solution, i.e.,
0.9% sodium chloride injection USP (available from B. Braun Medical
Inc.).
[0438] Groups of dogs from the Control and High level 14-day
exposures were also studied following a 14-day recovery period.
Endpoints for all groups of dogs included physical examinations,
clinical observations, body weights, opthalmology, cardiovascular
EKG, clinical pathology (hematology, clinical chemistry),
urinalyses, organ weights, histopathology, and toxicokinetics.
[0439] Vancomycin aerosol concentrations were 1.39.+-.0.20,
1.51.+-.0.19, and 1.49.+-.0.15 mg/L for the Low, Mid, and High
exposure levels, respectively. Mean particle size was determined to
be in the inhaleable size range for dogs (1.9-2.6 .mu.m mass median
aerodynamic diameter). Mean total inhaled doses were estimated as
10, 23, and 45 mg/kg, and mean doses deposited in lung were
estimated as 2, 5, and 9 mg/kg for the Low, Mid, and High exposure
levels, respectively.
[0440] The vancomycin exposures were well-tolerated by all groups
of dogs. All dogs survived to scheduled necropsy. There were no
vancomycin related effects noted on physical examinations, clinical
observations, opthalmology, cardiac ECG tracings, hematology,
clinical chemistry, urinalyses, gross necropsy observations, and
organ weights.
[0441] Histopathology examinations of tissues revealed no effect of
Vancomycin exposure in the organs and tissues examined outside of
the respiratory tract. Likewise, there was an absence of
microscopic alterations in the nasal cavity/turbinates, larynx, and
trachea. The effects of Vancomycin exposure were limited to
microscopic findings in the lung. Treatment-related increased
incidence of minimal to mild chronic interstitial inflammation,
alveolar histiocytosis, and bronchial lymph node lymphoreticular
hyperplasia were observed. Among Control and High level animals in
the Recovery groups there were no treatment-related differences in
the macroscopic and microscopic findings.
[0442] In conclusion, effects of Vancomycin exposure were limited
to minimal to mild pulmonary histopathology at the termination of
exposure. Recovery of histopathological effects was complete after
14 days. The minimal to mild chronic interstitial inflammation was
generally comparable with background inflammatory changes in beagle
dogs. The alveolar histiocytosis was reflective of enhanced
clearance that occurs without alveolar injury. The lymphoreticular
hyperplasia was considered an adaptive response that facilitates
lung clearance mechanisms. Since corresponding fibrosis and
alveolar epithelial injury were not characteristic of the observed
effects, the lung changes and related lymph node changes were not
considered adverse effects. Based on these findings, the no
observed adverse effect level (NOAEL) was the high exposure level
corresponding to an inhaled dose of 45 mg/kg and a deposited lung
dose of 9 mg/kg.
Example 7
[0443] Amikacin Sulfate sterile solution for inhalation, 125 mg/ml
was manufactured and characterized as follows. Approximately 13.5 L
of sterile water for injection was added to a glass carboy fitted
with a lightning labmaster mixer. Amikacin sulfate was added to the
carboy and the solution was stirred. The solution was mixed until
the entire API was dissolved. A sample of the solution was taken
and pH measured. With continued stirring, pH was adjusted with 1.0N
HCl to be within 5.5-6.3 with a target pH of 5.9. After pH
adjustment, sufficient quantity of sterile water for injection was
added until the final weight of solution of 21,328 g. was reached.
The pH of the final solution was verified to be within an
acceptable range. The solution was then sparged with filtered
nitrogen at a rate of 1.5 L/min for 15 minutes. The solution was
then filtered through the 0.22 micron sterile filter.
[0444] Prior to filling the solution, each vial was purged with
nitrogen. The solution was filled by weight using a Cazzoli
filler/stopper machine into 5 ml amber vials to a target weight of
4.27.+-.0.08 g. The vials were stoppered with 20 mm Teflon-coated
stoppers and secured with aluminum flip off seals. Filled vials
were stored at 2-8.degree. C. The composition is summarized in
Table A below.
TABLE-US-00033 TABLE A Ingredient g per batch Amikacin Sulfate
3525.0 g Hydrochloric Acid qs to pH 5.9 NaOH qs to pH 5.9 Sterile
Water for Injection qs to 21, 328 g Nitrogen, NF Qs
[0445] Stability over time was assessed for as formulation made
substantially as show in table A, with regard to total amikacin
active, related substances, such as degradation products,
appearance, pH, particulates and sterility. Thus samples were
stored at 5.degree. C. (Table B), at 25.degree. C./60% relative
humidity (RH) (Table C), and at 40.degree. C./75% RH (Table D). In
each case samples were stored in 5 mL amber glass vials, with 20 mm
Teflon stoppers and 20 mm aluminum overseals. Results of each of
these storage conditions are shown in Tables B, C and D,
respectively.
TABLE-US-00034 TABLE B Attributes Specification Initial 1 mo. 3
mos. 6 mos. 9 mos. 12 mos. 18 mos. 90.0%-110.0% l.s. 99.8 104.6
104.0 104.3 104.9 100.0 105.5 99.8 100.6 Appearance Meets
Test.sup.1 MT MT MT MT MT MT MT Kanamycin @ rrt Report results 0.73
0.78 0.68 0.34 0.33 0.28 0.22 0.72 0.64 Rel. Substance A Report
results 0.66 0.70 0.19 0.55 0.46 0.50 0.36 @ rrt 0.86 0.60
Unidentified @ rrt Report results 6.90 6.81 5.38 3.38 3.02 2.90
2.29 0.61 6.27 Unidentified @ rrt Report results 0.53 0.45 0.45
0.23 0.26 0.21 0.23 0.67 0.50 Total Related 10.0% 8.82 8.74 6.70
4.50 4.07 3.89 3.10 Substances 8.01 pH 5.5-6.3 5.6 5.6 5.5 5.6 5.5
5.7 5.6 Particulate Matter Particles = 10 .mu.m 50 3 7 24 38 5 2
NMT 6000 Particles = 25 .mu.m 0 0 0 0 1 0 0 NMT 600 Sterility Meets
USP Conforms NP NP NP NP MT NP
TABLE-US-00035 TABLE C Attributes Specifications Initial 1 mo. 3
mos. 6 mos. Assay 90.0%-110.0% 99.8 104.0 105.3 101.1 l.s. 99.8
Appearance Meets Test.sup.1 MT MT MT MT Kanamycin Report results
0.73 0.81 0.71 0.44 @ rrt 0.72 0.64 Rel. Substance Report results
0.66 0.72 0.18 0.59 A @ rrt 0.86 0.60 Unidentified Report results
6.90 6.87 5.28 3.55 @ rrt 0.61 6.27 Unidentified Report results
0.53 0.50 0.49 0.28 @ rrt 0.67 0.50 Total Related 10.0% 8.82 8.90
6.66 4.86 Substances 8.01 pH 5.5-6.3 5.6 5.6 5.6 5.6 Particulate
Particles = 50 12 18 33 Matter 10 .mu.m NMT 6000 Particles = 0 1 0
1 25 .mu.m NMT 600 Sterility Meets USP Conforms NP NP NP
TABLE-US-00036 TABLE D Attributes Specifications Initial 1 mo. 3
mos. 6 mos. Assay 90.0%-110.0% 99.8 104.1 104.7 106.3 l.s. 99.8
Appearance Meets Test.sup.1 MT MT MT clear, faint yellow solution
Kanamycin Report results 0.73 0.99 1.06 1.00 @ rrt 0.72 0.64 Rel.
Report results 0.66 0.71 0.16 0.60 Substance 0.60 A @ rrt 0.86
Unidentified Report results 6.90 6.81 4.64 3.52 @ rrt 0.61 6.27
Unidentified Report results 0.53 0.54 0.60 0.61 @ rrt 0.67 0.50
Total 10.0% 8.82 9.05 6.46 5.73 Related 8.01 Substances pH 5.5-6.3
5.6 5.6 5.5 5.5 Particulate Particles = 50 32 20 27 Matter 10 .mu.m
NMT 6000 Particles = 0 1 0 1 25 .mu.m NMT 600 Sterility Meets USP
Conforms NP NP NP
[0446] FIG. 16 is a graphical representation of certain of the
stability data provided in Tables B, C and D. In the Fig., the line
marked by diamonds represents the 5.degree. C. storage condition,
the line marked by the squares represents 25.degree. C./60% RH
storage conditions and the line marked by the triangle represents
40.degree. C./75% RH storage conditions. The Fig shows that the
percentage related substances, i.e. impurities, diminishes over
storage time. It is thought that this is a function of
detactability of the impurities. It is evident, however, that the
compositions remain stable, with respect to impurities, over
time.
[0447] Having now fully described this invention, it will be
understood to those of ordinary skill in the art that the methods
of the present invention can be carried out with a wide and
equivalent range of conditions, formulations, and other parameters
without departing from the scope of the invention or any
embodiments thereof.
[0448] All patents and publications cited herein are hereby fully
incorporated by reference in their entirety. The citation of any
publication is for its disclosure prior to the filing date and
should not be construed as an admission that such publication is
prior art or that the present invention is not entitled to antedate
such publication by virtue of prior invention.
* * * * *