U.S. patent application number 12/980477 was filed with the patent office on 2011-06-30 for rapid reconstitution for lyophilized-pharmaceutical suspensions.
This patent application is currently assigned to BAXTER INTERNATIONAL INC.. Invention is credited to Wei Y. Kuu.
Application Number | 20110155620 12/980477 |
Document ID | / |
Family ID | 44146708 |
Filed Date | 2011-06-30 |
United States Patent
Application |
20110155620 |
Kind Code |
A1 |
Kuu; Wei Y. |
June 30, 2011 |
RAPID RECONSTITUTION FOR LYOPHILIZED-PHARMACEUTICAL SUSPENSIONS
Abstract
A method of preparing and reconstituting a sterile, lyophilized
pharmaceutical active for rapid reconstitution by evacuating a
lyophilized pharmaceutical active-containing container until the
pressure within the container is less than about 300 Torr and
hermetically sealing the evacuated container. The sterile,
lyophilized pharmaceutical active can be prepared by flash freezing
a pharmaceutical active-containing composition then lyophilizing
the composition. The hermetically sealed lyophilized pharmaceutical
active can be reconstituted by adding at least the total volume of
liquid necessary for reconstitution of the sterile, lyophilized
pharmaceutical active to the sterile, lyophilized pharmaceutical
active, sealed under a pressure of less than about 300 Torr, in
less than about 10 seconds to yield, within about 5 minutes, an
administrable pharmaceutical active-containing composition. One
aspect of the herein described sterile, lyophilized pharmaceutical
active is a packaged sterile pharmaceutical active comprising an
evacuated, hermetically sealed container having disposed therein a
sterile, lyophilized pharmaceutical active, sealed under a pressure
of less than about 300 Torr.
Inventors: |
Kuu; Wei Y.; (Libertyville,
IL) |
Assignee: |
BAXTER INTERNATIONAL INC.
Deerfield
IL
BAXTER HEALTHCARE S.A.
Glattpark (Opfikon)
|
Family ID: |
44146708 |
Appl. No.: |
12/980477 |
Filed: |
December 29, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61290998 |
Dec 30, 2009 |
|
|
|
Current U.S.
Class: |
206/524.1 ;
53/405 |
Current CPC
Class: |
A61K 38/38 20130101;
A61K 9/19 20130101; A61K 31/337 20130101 |
Class at
Publication: |
206/524.1 ;
53/405 |
International
Class: |
B65D 85/00 20060101
B65D085/00; B65B 31/00 20060101 B65B031/00 |
Claims
1. A method of preparing a sterile, lyophilized pharmaceutical
active for rapid reconstitution, comprising: evacuating a sterile,
lyophilized pharmaceutical active-containing container until the
pressure within the container is less than about 300 Torr (40 kPa);
and hermetically sealing the evacuated container.
2. The method of claim 1 further comprising: flash freezing a
sterile pharmaceutical active-containing composition; lyophilizing
the flash frozen sterile pharmaceutical active-containing
composition to yield the sterile, lyophilized pharmaceutical
active; and thereby preventing Ostwald ripening growth.
3. The method of claim 2, wherein the lyophilizing of the flash
frozen pharmaceutical active-containing composition comprises:
storing a flash frozen pharmaceutical active-containing composition
in a lyophilization chamber; warming the flash frozen
pharmaceutical active-containing composition disposed within the
lyophilization chamber to a temperature of less than about
-20.degree. C.; pulling a vacuum of less than about 1 Torr (133 Pa)
on the lyophilization chamber; warming the flash frozen
pharmaceutical active-containing composition disposed within the
lyophilization chamber to about -20.degree. C.; and maintaining the
vacuum of less than about 1 Torr (133 Pa) on the lyophilization
chamber for a period of time constituting a primary drying cycle
that corresponds with a removal of a majority of a frozen
solvent.
4. The method of claim 3 further comprising: after the end of the
primary drying cycle, then warming the pharmaceutical
active-containing composition to about 30.degree. C. and
maintaining the vacuum of less than about Ton (133 Pa) for a period
of time constituting a secondary drying cycle.
5. A method of reconstituting a sterile, lyophilized pharmaceutical
active comprising: adding a volume of a sterile liquid to a
sterile, lyophilized pharmaceutical active, sealed within an
evacuated container wherein the pressure within the evacuated
container is less than about 300 Torr (40 kPa), in less than about
10 seconds to yield, within about 5 minutes, an administrable
pharmaceutical active-containing composition; wherein the volume of
liquid added is at least the total volume of liquid necessary for
reconstitution of the sterile, lyophilized pharmaceutical
active.
6. The method of claim 5, wherein the sterile, lyophilized
pharmaceutical active is one having been lyophilized from a first
volume of a liquid containing the pharmaceutical active, and
wherein the volume of liquid added for reconstitution is equivalent
to the first volume.
7. The method of claim 5, wherein the liquid is a solvent for the
pharmaceutical active.
8. A packaged sterile pharmaceutical active comprising a sterile,
lyophilized pharmaceutical active hermetically sealed within an
evacuated container; wherein the pressure within the evacuated
container is less than about 300 Torr (40 kPa).
9. The package of claim 8, wherein the container is selected from
the group consisting of a rigid container, a non-rigid container,
and a variable-volume container.
10. The package of claim 8, wherein the sterile, lyophilized
pharmaceutical active comprises paclitaxel.
11. The package of claim 8, wherein the sterile, lyophilized
pharmaceutical active comprises an albumin.
12. The package of claim 11, wherein the pharmaceutical active
comprises at least about 50 wt. % of the albumin and wherein the
albumin is a human serum albumin.
13. The package of claim 12, wherein the pharmaceutical active
comprises at least about 75 wt. % of the human serum albumin.
14. The package of claim 13, wherein the pharmaceutical active
comprises at least about 85 wt. % of the human serum albumin.
15. The package of claim 8, wherein the sterile pharmaceutical
active-containing composition comprises a nanoparticle of human
serum albumin.
16. The package of claim 8, wherein the pressure within the
evacuated container is less than about 100 Torr (13 kPa).
17. The package of claim 16, wherein the pressure is less than
about 25 Torr (3325 Pa).
18. The package of claim 17, wherein the pressure is less than
about 5 Torr (667 Pa).
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] The benefit of U.S. Provisional Patent Application
61/290,998, filed Dec. 30, 2009, is hereby claimed, and the
disclosure thereof is incorporated herein by reference in its
entirety.
FIELD OF THE DISCLOSURE
[0002] The invention relates to the pharmaceutical and medical
field and provides a packaged lyophilized pharmaceutical active and
a method of producing the packaged lyophilized pharmaceutical
active. Specifically, the invention provides for the rapid
reconstitution of a lyophilized pharmaceutical active with minimal
foaming.
BACKGROUND
[0003] Lyophilization is used to prepare pharmaceutical actives
with higher stability, broader temperature tolerance, and longer
shelf-life than comparable aqueous solutions. Typically,
pharmaceutical actives are dried to water contents of less than 5
wt. % through sublimation and desorption.
[0004] The reconstitution of lyophilized pharmaceutical actives
that contain proteins or polypeptides by the addition of water,
leads to the formation of a foam. The foam is thought tooccur from
the interaction of the amphiphilic protein or polypeptide with the
reconstituting solvent, typically water. The foaming of the
lyophilized pharmaceutical active poses problems for medical
practitioners and researchers. For example, in double blind studies
placebos of lyophilized pharmaceuticals need to appear identical to
the active therefore a placebo must foam identically to the active.
See e.g., U.S. Pat. No. 6,242,423. The reduction or elimination of
foaming would benefit both the medical practitioner by saving time
and the researcher by reducing the complication of formulating a
placebo for a double blind study.
SUMMARY OF THE INVENTION
[0005] Described herein, in the preferred embodiment, is a
lyophilized pharmaceutical active stored under a pressure of less
than 300 Torr (40 kPa) that can be rapidly reconstituted for in
vivo use.
[0006] One aspect of the materials and methods described herein is
a process for making a sterile, lyophilized pharmaceutical active.
This process includes evacuating a container that contains the
sterile, lyophilized pharmaceutical active and hermetically sealing
the container.
[0007] Another aspect of the materials and methods described herein
is a process for preventing Ostwald ripening growth during the
preparation of a sterile, lyophilized pharmaceutical active. This
process includes flash freezing a sterile pharmaceutical active
containing composition, lyophilizing the composition, and sealing
the sterile, lyophilized pharmaceutical active under a pressure of
less than about 300 Torr (40 kPa).
[0008] Yet another aspect of the materials and methods described
herein is a process for reconstituting a sterile, lyophilized
pharmaceutical active. This process includes adding a sterile
liquid to the sterile, lyophilized pharmaceutical active to yield
an administrable pharmaceutical active-containing composition
within about 5 minutes.
[0009] Still another aspect of the materials and methods described
herein is a packaged sterile pharmaceutical active. This package
includes a sterile, lyophilized pharmaceutical active hermetically
sealed within a container.
DETAILED DESCRIPTION
[0010] Herein, ranges may be expressed as from "about" or
"approximately" one particular value and/or to "about" or
"approximately" another particular value. When such a range is
expressed, another embodiment includes from the one particular
value and/or to the other particular value. Similarly, when values
are expressed as approximations, by use of the antecedent "about,"
it will be understood that the particular value forms another
embodiment. Additionally, compilations of parts of or areas in
devices are at times designated specific regions, these regions are
described based on the theorized primary event occurring in the
designated region. Regions can and often overlap and other events
can and likely occur within the specifically designated
regions.
[0011] The article and methods described herein generally relates
to the preparation, storage and reconstitution of pharmaceutical
actives. An aspect of the reconstitution of pharmaceutical actives
is the amount of time necessary for the reconstitution, with
multi-step and/or prolonged reconstitution processes being
burdensome on an administrating medical practitioner. Embodiments
of the herein described methods and article can be selected to
reduce the medical practitioner's time commitment to the
reconstitution process and thereby the medical practitioner can
more readily care for the patient.
[0012] One aspect of the herein described article and methods is
the preparation of a sterile, lyophilized pharmaceutical active
("LPA") for rapid reconstitution, comprising evacuating a
lyophilized pharmaceutical active-containing container until the
pressure within the container is less than about 25 Torr (3333 Pa);
and hermetically sealing the evacuated container. Another aspect of
the herein described methods is the prevention of Ostwald ripening
growth by flash freezing during the preparation of the sterile,
LPA. Still another aspect of the herein described methods is the
rapid reconstitution of the sterile, LPA. Yet another aspect of the
herein described article is the packaged sterile, LPA, for example,
prepared by any one of the methods described herein.
[0013] Generally, the preparation of the sterile, LPA described
herein involves the lyophilization and storage of a pharmaceutical
active-containing composition, often containing a solvent or
liquid, e.g., water. In addition, the pharmaceutical active can be
in the formof a solution, a suspension, or an emulsion. The
pharmaceutical active can be lyophilized by placing a
pharmaceutical active-containing composition in a freeze dryer or a
lyophilizer, and lyophilizing. One procedure for lyophilizing a
pharmaceutical active-containing composition includes continuously
pulling a vacuum (e.g. to less than about 1 Torr (133 Pa),
preferably about 0.01 to about 1 Torr, more preferably about 60 to
about 300 mTorr (8 to 40 Pa), even more preferably to about 100
mTorr (13 Pa) or less) on a pharmaceutical active-containing
composition that was precooled and maintained at a temperature of
less than about -20.degree. C., more preferably a temperature of
about -20.degree. C. to about -70.degree. C., for example a
temperature of about -40.degree. C.
[0014] A lyophilization of a pharmaceutical active-containing
composition can be divided into a plurality of drying cycles, e.g.,
the drying cycles can include a primary drying cycle and a
secondary drying cycle. The primary drying cycle can be a period of
time that depends on the removal of an amount of frozen solvent,
e.g., the majority, from the pharmaceutical active-containing
composition. The plurality of drying cycles can include a primary
to secondary drying cycle transition which can include raising the
temperature of the pharmaceutical active-containing composition,
e.g., at about 0.5.degree. C. per minute to about -20.degree. C.,
while continuously pulling a vacuum. The secondary drying cycle can
be a period of time that depends on the removal of an amount of
residual solvent in the pharmaceutical active-containing
composition, e.g., that not removed in the primary drying cycle, if
any, and which would be apparent to one of ordinary skill. The
plurality of drying cycles can further include a secondary to
tertiary drying cycle transition that can include raising the
temperature of the pharmaceutical active-containing composition, at
about 0.5.degree. C. per minute to about 30.degree. C., while
continuously pulling a vacuum. A tertiary drying cycle can be a
period of time, that is apparent to one of ordinary skill, that
includes continuously pulling vacuum at a temperature above the
melting point of a solvent that was included in the pharmaceutical
active-containing composition prior to lyophilization.
[0015] An added benefit can be obtained if Ostwald ripening growth
is prevented during the preparation of the sterile, LPA.
Preferably, the pharmaceutical active-containing composition is
flash frozen prior to lyophilization. The pharmaceutical
active-containing composition can be flash frozen by, for example,
submerging a container holding the composition in a cryogenic
liquid, e.g., liquid nitrogen. Alternative methods exist for the
flash freezing of the composition and will be available to one of
ordinary skill. Preferably, the composition is frozen at a
sufficient rate (e.g. within about 60 seconds) to prevent Ostwald
ripening growth. When the pharmaceutical active-containing
composition is cryogenically flash frozen the composition can be
subsequently warmed within the lyophilization chamber (e.g. to a
temperature of about -40.degree. C.) while exposed to a vacuum
(e.g. to less than about 1 Torr (133 Pa), preferably about 0.01 to
about 1 Torr, more preferably about 60 to about 300 mTorr (8 to 40
Pa), even more preferably to about 100 mTorr (13 Pa) or less).
[0016] Generally, the storage of the LPA involves disposing the LPA
in a sealable container and preferably sealing the container.
Preferably, after lyophilization, the LPA is disposed within a
hermetically sealable container; the container is evacuated,
preferably, to a pressure less than about 300 Torr, more preferably
less than about 200 Torr, even more preferably less than about 100
Torr, still more preferably less than about 50 Torr, even still
more preferably less than about 25 Torr (3333 Pa), yet more
preferably less than about 10 Torr (1333 Pa), and yet still more
preferably less than about 5 Torr (667 Pa) or 4 Torr (533 Pa); and
the container is then hermetically sealed. One of ordinary skill
will comprehend methods for storing a material within a container.
These methods include and are not limited by the methods of 1)
storing the LPA in a sealable container, evacuating the container,
and then hermetically sealing the container; 2) sealing the LPA in
a container, evacuating the container, and then applying a hermetic
seal; or 3) evacuating a chamber containing the LPA and the
container, and then hermetically sealing the LPA inside the
container. Optionally, the sealed container contains a dissolution
aid, for example a dissolution aid can be a impeller, a magnetic
stirrer, and/or other inert solids like glass beads, polyethylene
beads, and ceramic beads.
[0017] The sealable container can be a rigid container, a non-rigid
container, or a variable-volume container. Examples of rigid
containers include ampoules, vials, tubes, bottles, and the like;
for example made of glass, plastic, and the like. Non-rigid
containers include hermetically sealable plastic bags. When the
sealable container is a non-rigid container one of ordinary skill
would recognize that the pressure within the container after
equilibration is typically not the same pressure as when the
container was evacuated, e.g., the container deforms to equilibrate
the external pressure and the internal pressure; a non-rigid
container sealed under a pressure of less than about 25 Torr (3333
Pa) will equilibrate to a higher internal pressure, approaching or
achieving the external pressure. Variable-volume containers include
for example syringes. While syringes hermetically sealed under
vacuum may decrease the internal volume within the syringe,
syringes with barrel-locking mechanisms may be used to maintain a
minimum volume within the syringe and thereby a known internal
pressure. One of ordinary skill in the art will comprehend how to
hermetically seal the sealable container. Rigid containers are
preferred.
[0018] The methods described above will yield a packaged sterile,
LPA. Preferably, the packaged sterile, LPA comprises a LPA disposed
within a hermetically sealable container. More preferably, the LPA
is sealed within the container at a pressure of less than about 300
Torr, more preferably less than about 200 Torr, even more
preferably less than about 100 Torr, still more preferably less
than about 50 Torr, even still more preferably less than about 25
Torr (3333 Pa), yet more preferably less than about 10 Torr (1333
Pa), and yet still more preferably less than about 5 Torr (667 Pa)
or 4 Torr (533 Pa). One of ordinary skill in the art will recognize
that the pressure in the hermetically sealed container is dependent
on the rigidity of the container, therefore the pressure within the
container may be different from the pressure at which the container
was sealed.
[0019] The reconstitution of the pharmaceutical active by the
addition of a solvent for the pharmaceutical active is preferably
carried out under reduced pressure. One of ordinary skill in the
art will recognize that there are multiple methods for the addition
of a solvent to a material stored under a reduced pressure. Herein,
all methods for the addition of the solvent to the pharmaceutical
active are applicable so long as the addition occurs with minimum
addition of a gas. These methods include the cannulae transfer of
the solvent, the syringe transfer of the solvent, addition of the
solvent in a vacuum chamber, vacuum distillation of the solvent
into the pharmaceutical active containing container, and the like.
More preferably, the addition of the solvent occurs without the
formation of bubbles or foam in the pharmaceutical
active-containing container. Still more preferably, the solvent is
degassed prior to the addition to the pharmaceutical active.
[0020] The reconstitution of the pharmaceutical active is
preferably rapid. Herein, the reconstitution of known
pharmaceutical actives preferably occurs in 50% less time than the
time necessary under the current and accepted procedures for the
reconstitution of the same pharmaceutical actives. More preferably,
the reconstitution of the pharmaceutical active occurs as a single
step through the addition of the total volume of solvent necessary
for reconstitution of the pharmaceutical active. Preferably, the
total volume of solvent is added in less than about 60 seconds,
more preferably less than about 30 seconds, even more preferably
less than about 20 seconds, and still more preferably less than
about 10 seconds; thereafter the reconstituted pharmaceutical
active-containing composition is preferably administrable within
about 10 minutes, more preferably within about 5 minutes, and even
more preferably within about 2.5 minutes.
[0021] The pharmaceutical active is one that can be lyophilized and
reconstituted. In one embodiment, the pharmaceutical active
includes an albumin. Typically, lyophilized albumin compositions
foam upon reconstitution with water due to the interaction of the
amphiphilic albumin and the water. The foaming of lyophilized
albumin forces prolonged reconstitution procedures to prevent the
injection or biological application of the foamed albumin, thereby
preventing rapid reconstitution and administration. The foaming of
the reconstituted albumin is especially detrimental to the rapid
reconstitution and administration of pharmaceutical actives
intended for human use. When intended for human use the albumin is
preferably a human serum albumin. More preferably the
pharmaceutical active has nanoparticles of human serum albumin. The
amount of albumin in the pharmaceutical active can vary.
Preferably, the pharmaceutical active has at least about 50 wt. %
of the albumin, more preferably about 75 wt. %, and even more
preferably about 85 wt. %. When the pharmaceutical active comprises
an albumin the active can further comprise a drug or drugs that
bind to an albumin. Examples of drugs that bind to human serum
albumin are known to one of ordinary skill and include ontological,
imaging, anti-inflammatory, and antiallergic drugs. A preferable
ontological drug that binds to human serum albumin is
paclitaxel.
EXAMPLES
[0022] Several lyophilization and reconstitution studies were
performed to test the invention using: 1) ABRAXANE vials, 2)
homogenized 0.3% paclitaxel/albumin formulation, 3) 5% BSA (bovine
serum albumin), and 4) 3% BSA. ABRAXANE is a nanoparticle
suspension containing approximately 4.5% HSA (human serum albumin)
and 0.5% paclitaxel after reconstituted with 20 mL of the diluent.
Thus, 90% of the content of the ABRAXANE vials is HSA which is a
protein. As such, HSA contributes the major physical
characteristics, such as texture of the lyophilized vial,
reconstitution rate, and foaming, of the ABRAXANE product. Thus
placebo vials, i.e. vials containing HSA without paclitaxel, will
give a close representation of the above physical characteristics.
For convenience, BSA was used as the placebo for the testing, since
pure BSA in solid form is more available than HSA. The 5% BSA was
used to mimic the ABRAXANE with 20 mL of fill volume for
lyophilization, and 3% BSA was used as the placebo to mimic 33 mL
fill volume for lyophilization. The studies were performed per the
following experimental designs: [0023] 1) Reconstituted the
original ABRAXANE vial with 20 mL pure water and recorded the
reconstitution time following the procedure described in the
package insert of the product. [0024] 2) Pulled a vacuum on the
ABRAXANE vial, to approximately 4 Torr (533 Pa) using a LYOSTAR II
dryer, followed by reconstitution with 33 mL water to study the
effect of vacuum on the reconstitution rate. [0025] 3) Divided the
above reconstituted vial into 4 vials, 4 mL each into 10 mL vials,
followed by lyophilization and sealing under a high vacuum of 4
Torr (533 Pa) to 5 Torr (667 Pa). Studied the reconstitution time
of the resulting vials. The purpose of dividing the content in the
ABRAXANE vial was due to the high cost of ABRAXANE. The results
from these small vial studies gave a close indication of the
physical characteristics of the actual size of the ABRAXANE vial.
[0026] 4) Prepared 5% BSA and filled 4 mL each into 10 mL vials,
followed by lyophilization and reconstitution, and compared the
results with those in Step 2. [0027] 5) Lyophilized approximately
30 mL of 0.3% paclitaxel/albumin nanoparticle suspension in 50 mL
vials, followed by lyophilization and reconstitution. [0028] 6)
Prepared 3% BSA and filled 33 mL each into 50 mL vials, followed by
lyophilization and reconstitution, and compares the results with
those in Step 4.
[0029] The results of the above studies are presented below.
Example 1
Lyophilization and Reconstitution Studies for 5% BSA (Bovine Serum
Albumin), 5 mL Fill into 10 mL Vials
[0030] The purpose of using 4 mL fill of ABRAXANE in 10 mL vials
was due to the high cost of ABRAXANE. The small vial studies
generated more replicates for data analysis.
[0031] The lyophilization cycle parameters are: [0032] 1) Cool the
shelf to -40.degree. C. and hold for 80 minutes. [0033] 2) Pull
vacuum to 100 mTorr (133 Pa). [0034] 3) Hold at -40.degree. C., 100
mTorr (133 Pa) for 30 minutes. [0035] 4) Ramp 0.5.degree. C. to
-20.degree. C., 100 mTorr (133 Pa). [0036] 5) Primary drying: Hold
at -20.degree. C., 100 mTorr (133 Pa) for approximately 2300
minutes. [0037] 6) Ramp 1.degree. C./min to 30.degree. C., 100
mTorr (133 Pa). [0038] 7) Secondary drying: Hold at 30.degree. C.,
100 mTorr (133 Pa) for approximately 330 minutes.
[0039] After lyophilization, the vials were sealed at approximately
5 Torr (667 Pa). The reconstitution was performed by pointing the
needle at the cake to allow water to quickly absorbed by the
cake.
[0040] It was observed minimal amount of foam produced, probably
due to the fact that the vials were sealed at a high vacuum of
approximately 5 Torr (667 Pa). The vial was gently rotated by
fingers, and it was observed complete dissolution of the cake
approximately 4 minutes from the time when water was injected into
the vial.
Comparative Example 1
Reconstitution of the Original ABRAXANE Vial with 20 mL Water
[0041] The purpose of the study was to determine the reconstitution
time and foaming phenomenon of the ABRAXANE vial which contained
very little vacuum.
TABLE-US-00001 Reconstitution time Comment Approximately 20 min The
reconstitution procedure provided in the package insert was
carefully followed to avoid foaming.
[0042] As seen, approximately 20 minutes was required to
reconstitute the vial with a very careful control of the
reconstitution procedure described in the package insert.
Example 2
Lyophilization and Reconstitution Studies for Reconstituted
ABRAXANE Vial (4 mL Filled into 10 mL vials) and 5% BSA (5 mL Fill
into 10 mL Vials)
[0043] The purpose of the study was to investigate the
lyophilization and reconstitution characteristics of the
reconstituted ABRAXANE vials and the placebo vials of 5% BSA.
[0044] The lyophilization cycle parameters are: [0045] 1) Cool the
shelf to -40.degree. C. and hold for 90 minutes. [0046] 2) Pull
vacuum to 100 mTorr (133 Pa). [0047] 3) Hold at -40.degree. C., 100
mTorr (133 Pa) for 20 minutes. [0048] 4) Ramp at 0.5.degree. C./min
to -20.degree. C., 100 mTorr (133 Pa). [0049] 5) Hold at
-20.degree. C., 100 mTorr (133 Pa) for approximately 2400 minutes.
[0050] 6) Ramp at 0.5.degree. C./min to 30.degree. C., 100 mTorr
(133 Pa). [0051] 7) Hold at 30.degree. C., 100 mTorr (133 Pa) for
approximately 300 minutes.
[0052] After completion of the cycle run, the vials were sealed at
approximately 4 Torr 533 Pa) and crimped. The results of
reconstitution are discussed below.
[0053] (1) Reconstitution Time of 4 mL of Reconstituted ABRAXANE
Vials
TABLE-US-00002 Sample number Reconstitution time Comments 1 3 min
35.41 sec foaming was minimal 2 3 min 36.05 sec foaming was minimal
3 3 min 34.72 sec foaming was minimal 4 2 min 41.89 sec foaming was
minimal 5 3 min 25.21 sec foaming was minimal
[0054] (2) Reconstitution Time for 5% BSA
TABLE-US-00003 Sample number Reconstitution time Comments 6 4 min
25.87 sec foaming was minimal 7 3 min 2.87 sec foaming was minimal
8 0 min 33.91 sec foaming was minimal 9 4 min 6.22 sec foaming was
minimal 10 1 min 44.75 sec foaming was minimal 11 3 min 46.03 sec
foaming was minimal 12 2 min 50.19 sec foaming was minimal
[0055] As seen, the reconstitution time for all vials are very
short.
[0056] (3) 5% BSA with Glass Beads
TABLE-US-00004 Sample number Reconstitution time Comments 13 1 min
39.75 sec foaming was minimal 14 1 min 55.41 sec foaming was
minimal 15 0 min 33.72 sec foaming was minimal 16 1 min 55.41 sec
foaming was minimal 17 1 min 36.34 sec foaming was minimal
[0057] The above reconstitution times are even shorter than those
of 5% BSA without glass beads in (2).
Example 3
Quick Freezing, Lyophilization and Reconstitution Studies for
Reconstituted ABRAXANE Vial (4 mL filled into 10 mL Vials) and 5%
BSA (5 mL Filled into 10 mL Vials)
[0058] The purpose of quick freezing was to immobilize
nanoparticles from aggregation or crystallization due to the
Ostwald ripening effect.
[0059] Quick Freezing Procedure: [0060] 1) Placed the test-tube
rack on a metal tray and placed apiece of aluminum foil on the
bottom of the test tube rack and lifted the four sides of the foil
to cover the bottom half of the rack for holding the liquid
nitrogen. [0061] 2) Placed the reconstituted vials in the slots of
the test tube rack. [0062] 4) Placed stoppers on the vials. [0063]
5) Poured liquid nitrogen into the test tube rack to quick freeze
the vials. [0064] 6) Started the LyoStar II freeze dryer to precool
the shelf to -40.degree. C.
[0065] 7) Loaded the frozen vials onto the shelf, and started the
following cycle.
[0066] Lyophilization Cycle Parameters: [0067] 1) Cool the shelf to
-40.degree. C. and hold for 90 minutes. [0068] 2) Pull vacuum to
100 mTorr (133 Pa). [0069] 3) Hold at -40.degree. C., 100 mTorr
(133 Pa) for 30 minutes. [0070] 4) Ramp at 0.5.degree. C./min to
-10.degree. C., 100 mTorr (133 Pa). [0071] 5) Primary drying: Hold
at -10.degree. C., 100 mTorr (133 Pa) for approximately 1050
minutes. [0072] 6) Ramp at 0.5.degree. C./min to 30.degree. C., 100
mTorr (133 Pa). [0073] 7) Secondary drying: Hold at 30.degree. C.,
100 mTorr (133 Pa) for approximately 530 minutes.
[0074] After completion of the cycle, the vials were sealed at
approximately 4 Torr (533 Pa) and crimped.
[0075] The Results of Reconstitution Studies are Presented
Below
[0076] (1) ABRAXANE Vials (Injected with 4 mL Pure Water)
TABLE-US-00005 Sample number Reconstitution time Comment 18 2 min
3.50 sec foaming was minimal 19 2 min 15.66 sec foaming was minimal
20 2 min 45.25 sec foaming was minimal
[0077] (2) 5% BSA Vials Without Glass Beads (Injected with 5 mL
Pure Water)
TABLE-US-00006 Sample number Reconstitution time Comments 21 59.94
sec foaming was minimal 22 2 min 49.84 sec foaming was minimal 23 4
min 6.45 sec foaming was minimal 24 1 min 47.66 sec foaming was
minimal 25 2 min 31.72 sec foaming was minimal 26 4 min 12.45 sec
floating cake taking longer to dissolve 27 3 min 35.35 sec foaming
was minimal
[0078] (3) 5% BSA with Glass Beads (Injected with 5 mL Pure
Water)
TABLE-US-00007 Sample number Reconstitution time Comments 28 51.84
sec foaming was minimal 29 57.22 sec foaming was minimal 30 2 min
57.03 sec foaming was minimal 31 3 min 49.00 sec foaming was
minimal
[0079] The above results indicate that the resulting cakes after
quick freezing followed by lyophilization also reconstituted
rapidly. Thus the combination of quick freezing and vacuum seal can
prevent aggregation (or crystallization) of nanoparticles and give
a quick reconstitution of the resulting cakes.
Comparative Example 3
Reconstitution of Vials Sealed Under Ambient Pressure
[0080] The purpose of this study was to investigate the
reconstitution time for samples prepared by the quick freezing and
lyophilization procedure described above wherein the samples were
sealed under ambient pressure, approximately 760 Torr (101
kPa).
[0081] (1) ABRAXANE Vials (Injected with 4 mL Pure Water)
TABLE-US-00008 Sample number Reconstitution time Comment 32 3 min
41.66 sec large amount of foaming
[0082] (2) 5% BSA Vials without Glass Beads (Injected with 5 mL
Pure Water)
TABLE-US-00009 Sample number Reconstitution time Comments 33 9 min
30.28 sec large amount of foaming
[0083] (3) 5% BSA with Glass Beads (Injected with 5 mL Pure
Water
TABLE-US-00010 Sample number Reconstitution time Comments 34 4 min
52.93 sec large amount of foaming
[0084] The above-described results indicate that the resulting
cakes, after quick freezing followed by lyophilization, cannot be
reconstituted rapidly if the samples are not sealed under vacuum.
As seen, the time necessary for reconstitution ranged from about
60% longer to about 260% longer when the vials were sealed without
vacuum compared to sealing under a pressure of approximately 4 Torr
(533 Pa).
Example 4
Vacuum Pulled on the Original ABRAXANE Vial, Followed by
Reconstitution with 33 mL Water
[0085] The purpose of the study was to investigate the effect of
vacuum on the reconstitution rate of the original ABRAXANE product
vial. The ABRAXANE vial was vacuum pulled to approximately 4 Torr
(533 Pa) using LyoStar II dryer and resealed. The purpose of vacuum
pull was to minimize foaming during reconstitution. The ABRAXANE
vial was then reconstituted with 33 mL of pure water.
TABLE-US-00011 Vial number Reconstitution time 35 Approximately 12
minutes and 1.9 sec.
[0086] The reconstitution was performed by quickly injecting water
through the stopper and pointing at the lumps. It was observed that
the reconstitution was shorter than the 20 minutes in Comparative
Example 1.
Example 5
Lyophilization and Reconstitution Studies for 0.3%
Paclitaxel/Albumin and 3% BSA
[0087] The 0.3% paclitaxel/albumin nanoparticle suspension was
prepared at Baxter Round Lake facility. The vials contained
approximately 30 mL each in 50 mL vials. The placebo of 3% BSA was
prepared at Baxter Bloomington facility. The fill volume of the
placebo was 33 mL each in 50 mL vials. The vials were lyophilized
together using the following cycle parameters: [0088] (1) Precool
the shelf to 5.degree. C. [0089] (2) Load the vials to the shelf
and install thermocouples. [0090] (3) Cool the shelf to -40.degree.
C. and hold for approximately 270 min. [0091] (4) Start vacuum pump
at 100 mTorr (133 Pa) and hold at -40.degree. C. for 30 min. [0092]
(5) Primary drying: ramp to -10.degree. C. at 0.5.degree. C./min,
at 100 mTorr (133 Pa), and hold for 135 hours and 37 minutes.
[0093] (6) Secondary drying: ramp to 30.degree. C. at 1.degree.
C./min, and hold for 10 hours.
[0094] After completion of the cycle, the vials were sealed at
approximately 4 Torr (533 Pa). The following procedure was used to
reconstitute the vials. [0095] (1) Removed the aluminum cap of the
vial. [0096] (2) Load the syringe with 20 mL o ater, and ensure all
air is removed by priming. [0097] (3) Stick the needle through the
stopper and point at the cake [0098] (4) The water in the syringe
will be automatically and quickly pulled into the vial due to the
high vacuum of approximately 4 Torr (533 Pa). [0099] (5) As soon as
all water is delivered, pull out the needle quickly to avoid
injecting air. [0100] (6) Gently swirl or invert the vial to
enhance dissolution of the cake.
[0101] As seen, the reconstitution procedure for the vacuum sealed
vials is very different than the ABRAXANE product vials, because of
foaming can be minimized by the high vacuum.
[0102] The following sample IDs are for 3% BSA to mimic the
ABRAXANE vials, since the solution in the vials became clear after
reconstitution and easier to observe the reconstitution and foam
dissipation. The reconstitution times for 3% BSA, 33 mL in 50 mL
vials, are presented below:
TABLE-US-00012 Sample ID Reconstitution time Comment 36 7 min 23.28
sec foaming was minimal 37 6 min 36.72 sec foaming was minimal 38
10 min 13.38 sec foaming was minimal 39 8 min 58.96 sec foaming was
minimal 40 4 min 07.53 sec foaming was minimal Crushed cake by
shaking 41 8 min 26.18 sec foaming was minimal
[0103] It was found that the crushed cake in sample 40
reconstituted quicker. The cake was crushed by hand, by end-to-end
shaking of the vial for a few minutes. The reconstitution times of
the intact cake, samples 36, 37, 38, 39, and 41, are still much
shorter than that of the ABRAXANE vial. This result further
confirmed the effectiveness of vacuum seal on enhancing the
reconstitution and minimizing foaming.
* * * * *