U.S. patent application number 15/925272 was filed with the patent office on 2018-07-26 for autoclavable suspensions of cyclosporin a form 2.
The applicant listed for this patent is Allergan, Inc.. Invention is credited to Wendy M. Blanda, Michelle Luu, David A. Marsh, Hongwen Ma Rivers.
Application Number | 20180207230 15/925272 |
Document ID | / |
Family ID | 47192237 |
Filed Date | 2018-07-26 |
United States Patent
Application |
20180207230 |
Kind Code |
A1 |
Blanda; Wendy M. ; et
al. |
July 26, 2018 |
AUTOCLAVABLE SUSPENSIONS OF CYCLOSPORIN A FORM 2
Abstract
Disclosed herein are autoclavable formulations of cyclosporin A
Form 2, methods of making such formulations, and methods of
treating diseases of the eye with such formulations.
Inventors: |
Blanda; Wendy M.; (Tustin,
CA) ; Rivers; Hongwen Ma; (San Marcos, CA) ;
Marsh; David A.; (Irvine, CA) ; Luu; Michelle;
(Anaheim, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Allergan, Inc. |
Irvine |
CA |
US |
|
|
Family ID: |
47192237 |
Appl. No.: |
15/925272 |
Filed: |
March 19, 2018 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15171866 |
Jun 2, 2016 |
9919028 |
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15925272 |
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13676362 |
Nov 14, 2012 |
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15171866 |
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61559849 |
Nov 15, 2011 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/32 20130101;
A61K 38/13 20130101; A61P 37/06 20180101; A61K 47/38 20130101; A61P
27/02 20180101; A61K 9/0048 20130101; C07K 7/645 20130101; A61K
47/36 20130101; C07K 1/306 20130101; A61P 37/08 20180101; A61P
27/04 20180101 |
International
Class: |
A61K 38/13 20060101
A61K038/13; C07K 7/64 20060101 C07K007/64; C07K 1/30 20060101
C07K001/30; A61K 9/00 20060101 A61K009/00; A61K 47/36 20060101
A61K047/36; A61K 47/32 20060101 A61K047/32; A61K 47/38 20060101
A61K047/38 |
Claims
1. A method of making a formulation of cyclosporin A, the method
comprising the steps of a) dissolving cyclosporin A form 2 in
solution; b) autoclaving the solution; c) adding a vehicle selected
from carboxymethyl cellulose, hydroxypropyl methyl cellulose,
hydroxyethyl cellulose, hyaluronic acid, polyvinylpyrrolidone,
Carbopol Ultrez 10NF, and Pluronic F127.
2. A method of making a formulation of cyclosporin A, the method
comprising the steps of a) dissolving in solution a vehicle
selected from carboxymethyl cellulose, hydroxypropyl methyl
cellulose, hydroxyethyl cellulose, hyaluronic acid,
polyvinylpyrrolidone, Carbopol Ultrez 10NF, and Pluronic F127; b)
adding to the solution cyclosporin A Form 2; c) autoclaving the
resulting mixture.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This patent application is a continuation of U.S. patent
application Ser. No. 15/171,866, filed Jun. 2, 2016, which is a
divisional of U.S. patent application Ser. No. 13/676,362, now
abandoned, which claims priority to U.S. Provisional Patent
Application No. 61/559,849, filed Nov. 15, 2011, the entire
contents of which are hereby incorporated by reference.
BACKGROUND
[0002] Aseptic processing of cyclosporin A suspensions in a
hyaluronic acid media (a hydrogel used as a suspending agent), is
complicated by the fact that both the drug and the hyaluronic acid
need to be pre-sterilized. Pre-sterilized hyaluronic acid is
extremely expensive, costing roughly $1 million dollars for a few
kilograms (roughly $10,000 per ounce) of sterile raw material.
Additionally, in the process of pre-sterilizing cyclosporin A, the
drug is degraded upon irradiation, as shown below and in FIGS. 1
and 2:
TABLE-US-00001 TABLE 1 Impact of Irradiation on Cyclosporin
Stability Sterilization Form 1 CsA Form 2 CsA Form 3 CsA Amorph.
CsA Mode (Potency and Imp.) (Potency and Imp.) (Potency and Imp.)
(Potency and Imp.) None 98.4% w/w 94.6% w/w 97.7% w/w 96.5% w/w
Total Imp: 0.6% Total Imp: 0.6% Total Imp: 0.8% Total Imp: 0.7% 15
kGy Gamma 93.9% w/w 91.8% w/w 94.3% w/w 92.1% w/w % Rel. Change
4.5% % Rel. Change 2.9% % Rel. Change 3.6% % Rel. Change 4.6% Total
Imp: 1.7% Total Imp: 1.8% Total Imp: 1.3% Total Imp: 1.4% 33 kGy
Gamma 90.7% w/w 88.5% w/w 91.0% w/w 87.7% w/w % Rel. Change 7.8% %
Rel. Change 6.4% % Rel. Change 6.9% % Rel. Change 9.2% Total Imp:
2.8% Total Imp: 2.4% Total Imp: 2.3% Total Imp: 2.3% E-Beam 92.6%
w/w 90.3% w/w 93.4% w/w 92.0% w/w % Rel. Change 5.9% % Rel. Change
4.6% % Rel. Change 4.5% % Rel. Change 4.7% Total Imp: 1.5% Total
Imp: 1.7% Total Imp: 1.6% Total Imp: 1.3%
Cooling the cyclosporin during irradiation does not significantly
improve the results, as shown in Table 2, below:
TABLE-US-00002 Impact on Cyclosporin Stability after irradiation
under Cold Conditions Sterilization Form 1 CsA Form 2 CsA Form 3
CsA Amorph. CsA Mode (Potency and Imp.) (Potency and Imp.) (Potency
and Imp.) (Potency and Imp.) None 99.4% w/w 97.6% w/w 98.4% w/w
96.5% w/w Total Imp: 0.7% Total Imp: 0.5% Total Imp: 0.7% Total
Imp: 0.7% Cold E-beam 94.6% w/w 91.1% w/w 94.6% w/w 92.3% w/w %
Rel. Change 4.8% % Rel. Change 6.7% % Rel. Change 3.9% % Rel.
Change 4.4% Total Imp: 1.5% Total Imp: 1.5% Total Imp: 1.8% Total
Imp: 1.3% Regular E-Beam % Rel. Change 5.9% % Rel. Change 4.6% %
Rel. Change 4.5% % Rel. Change 4.7% (from Previous Total Imp: 1.5%
Total Imp: 1.7% Total Imp: 1.6% Total Imp: 1.3% Study) % Relative
Change in Potency on Sterilization
Additional levels of degradants need to be qualified in preclinical
safety studies. Moreover, a suspension, prepared with only 90-95%
of the labeled Cyclosporin A (due to the pre-sterilization
process), has a substantial probability of failure to meet
regulatory guidelines for shelf-life, since regulatory authorities
generally prohibit shelf-lives below 90% of label.
[0003] The present invention solves these problems. Disclosed
herein are formulations of cyclosporin A, combined with a
parenterally-biocompatible suspending agent, which are sterile,
exceptionally stable to heat sterilization, and have excellent
long-term stability.
BRIEF DESCRIPTION OF THE DRAWINGS
[0004] FIGS. 1 and 2 show change in cyclosporin A potency with
change in crystal form and sterilization method.
[0005] FIG. 3 shows x-ray powder diffraction pattern data of
cyclosporin A Form 2 after autoclaving.
[0006] FIG. 4 shows congestion seen on slit lamp examination with
eight different formulations.
[0007] FIG. 5 depicts characteristic X-ray powder diffraction
(XRPD) patterns of CsA in a new crystalline form (designated as
Form 2 herein), tetragonal form (designated as Form 1 herein), and
orthorhombic form (designated as Form 3 herein).
[0008] FIG. 6 depicts the XRPD diffractogram of CsA crystalline
Form 2.
[0009] FIG. 7 depicts the water sorption/desorption profile of CsA
Form 2.
[0010] FIG. 8 depicts MDSC analysis of CsA Form 2 recovered from
0.04% formulation with 1% PS80.
[0011] FIG. 9 shows gross ocular congestion after an injection of
100 ul of CMC, HEC, HPMC, Pluronic and PVP in in phosphate buffered
saline was administered subconjunctivally to New Zealand white
rabbits. The rabbits were observed for seven days.
[0012] FIG. 10 shows gross ocular discharge in the experiment
described in FIG. 9.
[0013] FIG. 11 shows gross ocular swelling in the experiment
described in FIG. 9.
[0014] FIG. 12 shows the simulated XRPD pattern of cyclosporine A
forms.
DETAILED DESCRIPTION
Cyclosporin A
[0015] Cyclosporin A (CsA) is a cyclic peptide having the following
chemical structure:
##STR00001##
Its chemical name is
cyclo[(E)-(2S,3R,4R)-3-hydroxy-4-methyl-2-(methylamino)-6-octenoyl]-L-2-a-
minobutyryl-N-methylglycyl-N-methyl-L-leucyl-L-valyl-N-methyl-L-leuc
yl-L-alanyl-D-alanyl-N-methyl-L-leucyl-N-methyl-L-leucyl-N-methyl-L-valyl-
]. It is also known by the names cyclosporin, cyclosporine A,
ciclosporin, and ciclosporin A. It is the active ingredient in
Restasis.RTM. (Allergan, Inc., Irvine, Calif.), an emulsion
comprising 0.05% (w/v) cyclosporin. Restasis.RTM. is approved in
the United States to increase tear production in patients whose
tear production is presumed to be suppressed due to ocular
inflammation associated with keratoconjunctivitis sicca.
Cyclosporin A Form 2
[0016] Cyclosporin A is known to exist in an amorphous form, liquid
crystal form, tetragonal crystalline form (form 1), and an
orthorhombic form (form 3). A new crystalline form, cyclosporin A
Form 2, has recently been discovered.
[0017] The XRPD pattern of CsA Form 2 differs significantly from
the tetragonal form and orthorhombic form (FIG. 1). The major
crystalline peaks for CsA form 2 appear at (2.theta.) when scanned
by an X-ray diffractometer with X-ray source as Cu K.alpha.
radiation, .lamda.=1.54 .ANG., at 30 kV/15 mA: 7.5, 8.8, 10.2,
11.3, 12.7, 13.8, 14.5, 15.6 and 17.5 (d-spacing in crystal lattice
at about 11.8, 10.0, 8.7, 7.8, 7.0, 6.4, 6.1, 5.6 and 5.1 .ANG.,
respectively, FIG. 2). These major peaks are defined as those being
unique to Form 2 relative to the orthorhombic or tetragonal forms;
as well as, peaks having an intensity greater than 5 times the
background.
[0018] In one embodiment, the new crystalline form (Form 2) of CsA
is a nonstoichiometric hydrate of Cyclosporin A. In another
embodiment, the crystalline Form 2 is represented by the
formula:
##STR00002##
wherein X is the number of molecules of water and varies from 0 to
3. In one embodiment, X in the above formula is 2.
[0019] Form 2 appears to be a kinetically stable form of CsA in
aqueous suspensions. Suspensions containing Form 2 show no
conversion to other known polymorphic or pseudomorphic forms upon
storage. It has been found that Form 1 and the amorphous form
convert to Form 2 in the presence of water.
[0020] The single crystal structure of the hydrate form of CsA Form
2 has been determined and the crystal structure parameters are
listed in Table 2. These results indicate that Form 2 is unique
compared to other known crystalline forms of cyclosporine A.
TABLE-US-00003 TABLE 1 Crystal data and data collection parameters
of crystal structure solution of CsA Form 2. formula
C.sub.62H.sub.115N.sub.11O.sub.14 formular weight 1236.67 space
group P 2.sub.1 2.sub.1 2.sub.1 (No.19) a (.ANG.) 12.6390(5) b
(.ANG.) 19.7852(8) c (.ANG.) 29.568(2) volume (.ANG..sup.3)
7383.8(7) Z 4 d.sub.calc (g cm.sup.-3) 1.114 crystal dimensions
(mm) 0.27 .times. 0.18 .times. 0.12 temperature (K) 150 radiation
(wavelength in .ANG.) Cu K.sub.3 (1.54184) monochromator confocal
optics linear abs coef (mm.sup.-1) 0.640 absoprtion corretion
applied empirical.sup.3 transmission factors (min, max) 0.80, 0.93
diffractometer Rigaku RAPD-II h, k, l range -13 to 13 -21 to 21 -32
to 21 2.theta. range (deg) 5.38-115.00 mosaicity (deg) 1.31
programs used SHELXTL F.sub.0CO 2704.0 weighting
1/[o.sup.2(Fo.sup.2) + (0.0845P).sup.3 + 0.000DP] where P =
(Fo.sup.3 + 2Fc.sup.2)/3 data collected 37360 unique data 9964
R.sub.int 0.077 data used in refinement 9964 cutoff used in
R-factor calculations F.sub.o.sup.2 > 2.0s(F.sub.o.sup.2) data
with I > 2.0s(I) 6597 number of variables 834 largest shift/esd
in final cycle 0.00 R(F.sub.o) 0.061 R.sub.w(F.sub.o.sup.2) 0.145
goodness of fit 1.037 absolute structure determination Flack
parameter.sup.b (0.0(3))
The asymmetric unit of this CsA Form 2 contains one cyclosporine A
molecule and two water molecules. It is possible that any small
molecule that can hydrogen bond to water could play the role of
space filler, which would give a range of potential structures
running from the orthorhombic dihydrate to distorted monoclinic
dihydrate The XRPD pattern calculated from the single-crystal
structure is shown in FIG. 12 and it matches the experimental
pattern shown in FIG. 2. These matching patterns further
corroborate that Form 2 is a unique and pure crystalline form of
cyclosporine A.
[0021] Without wishing to be bound by theory, thermogravimetric
analysis combined with KF titration and vapor sorption desorption
analysis (VSA) suggest that CsA Form 2 is a non-stoichiometric
hydrate of CsA. The vapor sorption analysis of Cyclosporine Form 2
indicates that water content in the new crystal form reversibly
varies with relative humidity as shown in FIG. 7. Similar to the
tetragonal form, the new CsA form undergoes a phase transition to a
liquid crystal or amorphous form at 124.4.degree. C. prior to
melting as indicated by the modulated differential calorimetric
(MDSC) analysis (FIG. 8).
[0022] Cyclosporin A Form 2 may be obtained by suspending amorphous
0.05% cyclosporin A (w/v) in 1% A Polysorbate 80, heating the
solution to 65.degree. C., holding it at that temperature for 24
hours, and then recovering the precipitate by vacuum filtration.
One can then use the cyclosporin A Form 2 thus obtained to generate
additional amounts, using Cyclosporin A Form 2 as a seed crystal;
in this method, one suspends about 30 g cyclosporin A in a solution
of 900 ml water containing 1% (w/v) Polysorbate 80, heats the
solution to 65.degree. C., and then seeds it with 0.2 g of
cyclosporin A Form 2 at a temperature of 52.degree. C. The solution
is then stirred for about 22 hours at a temperature of between
about 61.degree. C. and 65.degree. C., and then recovers the
precipitate that results.
[0023] Further details regarding CsA Form 2 may be found in U.S.
patent application Ser. No. 13/480,710, the entire contents of
which are incorporated by reference herein.
Heat-Stable, Heat-Sterilized Suspensions of Cyclosporin A Form
2
[0024] Compositions of the invention are ophthalmically acceptable
suspensions of Cyclosporin A form 2. By "ophthalmically
acceptable," the inventors mean that the suspensions are formulated
in such a way as to be non-irritating when administered to the eye
of a mammal, such as a human.
[0025] The suspensions of the invention comprise cyclosporin A form
2 and a vehicle comprising a suspending agent such as hyaluronic
acid, a cellulose, polyvinylpyrrolidone (PVP), Pluronic.RTM.
copolymers based on ethylene oxide and propylene oxide, and
Carbopol.RTM. polymers.
[0026] In one embodiment, the suspension comprises cyclosporin A
Form 2 at a concentration of about 0.001% to about 10% (w/v). In
one embodiment, the suspension comprises cyclosporin A form 2 at a
concentration of about 0.001% (w/v) to about 0.01%, about 0.001%
(w/v) to about 0.04% (w/v), about 0.001% (w/v) to about 0.03%
(w/v), about 0.001% (w/v) to about 0.02% (w/v), or about 0.001%
(w/v) to about 0.01% (w/v). In another embodiment, the suspension
comprises cyclosporin A form 2 at a concentration of about 0.01%
(w/v) to about 0.05%, about 0.01% (w/v) to about 0.04% (w/v), about
0.01% (w/v) to about 0.03% (w/v), about 0.01% (w/v) to about 0.02%
(w/v), or about 0.01% (w/v) to about 0.01% (w/v). In another
embodiment, the suspension comprises cyclosporin A form 2 at a
concentration of about 0.01% (w/v) to about 0.1%, about 0.1% (w/v)
to about 0.5% (w/v), about 0.01% (w/v) to about 1% (w/v), or about
1% (w/v) to about 10%.
[0027] For example, the suspensions may comprise about 0.001%
(w/v), about 0.002% (w/v), about 0.003% (w/v), about 0.004% (w/v),
about 0.005% (w/v), about 0.006% (w/v), about 0.007% (w/v), about
0.008% (w/v), about 0.009% (w/v), about 0.01% (w/v), about 0.015%
(w/v), about 0.02% (w/v), about 0.025% (w/v), about 0.03% (w/v),
about 0.035% (w/v), about 0.04% (w/v), about 0.045% (w/v), about
0.05% (w/v), about 0.055% (w/v), about 0.06% (w/v), about 0.065%
(w/v), about 0.07% (w/v), about 0.075% (w/v), about 0.08% (w/v),
about 0.085% (w/v), about 0.09% (w/v), about 0.095% (w/v), about
0.1% (w/v), about 0.15% (w/v), about 0.2% (w/v), about 0.25% (w/v),
about 0.3% (w/v), about 0.35% (w/v), about 0.4% (w/v), about 0.45%
(w/v), about 0.5% (w/v), about 0.55% (w/v), about 0.6% (w/v), about
0.65% (w/v), about 0.7% (w/v), about 0.75% (w/v), about 0.8% (w/v),
about 0.85% (w/v), about 0.9% (w/v), about 0.95% (w/v), or about
1.0% (w/v) cyclosporin A form 2.
[0028] Examples are provided in Table 3, below:
TABLE-US-00004 TABLE 3 Autoclavable suspensions of cyclosporin A
Form 2. Autoclave CsA Gelling Conditions Formu- (Crystal CsA
Gelling Agent Agent (Temp lation form) (%) (Type) (%)
(.degree.C)/min) 1 2 20 CMC 5 121/10 2 3 20 CMC 3 121/10 3 NA 0
Carbopol 1.5 121/15 Ultrez 10 4 NA 0 Carbopol 2.0 121/15 Ultrez 10
5 NA 0 Carbopol 2.5 121/15 Ultrez 10 6 NA 0 Carbopol 1.0 121/15
Ultrez 10 7 NA 0 Carbopol 4.0 121/15 Ultrez 10 8 2 5 CMC 3 121/15 9
2 5 CMC 2 121/15 10 2 20 CMC 10 121/15 11 2 0 CMC 10 121/15 12 2 5
HPMC 3 121/15 13 2 5 HPMC 6 121/15 14 2 20 HPMC 6 121/15 15 2 20
HPMC 10 121/15 16 2 5 HPMC 6 121/15 17 2 20 HPMC 3 121/15 18 2 5
HPMC 3 121/15 19 2 20 HPMC 3 121/15 20 2 10 HPMC 4.5 121/15 21 2 10
HPMC 4.5 121/15 22 2 10 HEC 3 121/15 23 2 10 HEC 3 121/15 24 2 30
HEC 1 121/15 25 2 10 HA 3.5 121/15* 26 2 10 HA 2.5 121/15 27 2 30
HEC 1 121/15 28 2 30 HA 1 121/15* 29 2 10 HA 2.5 121/15 30 2 10 HA
3.5 121/15 31 2 10 HA 4.5 121/15 32 2 30 HA 3.0 121/15 33 2 20 HA
1.5 121/15 34 2 20 HA 2.5 121/15 35 2 20 HA 3.5 121/15 36 2 10 HA 4
121/15, 121/30, and 123/15 37 2 10 HA 4 121/15, 121/30, and 123/15
38 2 10 HA 4 121/15, 121/30, and 123/15 39 2 35 HA 1 121/15* 40 2 5
HA 3.5 121/15* 41 2 10 HA 3.5 121/15* 42 2 20 HA 2.0 121/15* 43 2
20 HA 2.0 121/15* 44 2 10 HA 3.5 121/15* 45 2 10 HA 3.5 121/15* 46
2 25 N/A 0 120/15 47 2 25 N/A 0 118/20 48 2 25 N/A 0 120/12 HEC1 2
5 HEC 5 121/15 HEC2 2 20 HEC 5 121/15 HEC3 2 5 HEC 2 121/15 HEC4 2
20 HEC 2 121/15 HEC5 2 5 HEC 5 121/15 HEC6 2 20 HEC 5 121/15 HEC7 2
5 HEC 2 121/15 HEC8 2 20 HEC 2 121/15 HEC9 2 10 HEC 3 121/15 PVP1 2
10 PVP 25 121/15 PVP2 2 10 PVP 25 121/15 PVP3 2 10 PVP 15 121/15
PVP4 2 10 PVP 15 121/15 PVP5 2 25 PVP 25 121/15 PVP6 2 25 PVP 25
121/15 PVP7 2 25 PVP 15 121/15 PVP8 2 25 PVP 15 121/15 PVP9 2 10
PVP 25 121/15 PVP10 2 25 PVP 25 121/15 CsA = cyclosporin A. CMC =
carboxymethyl cellulose. HPMC = hydroxypropyl methyl cellulose. HEC
= hydroxyethyl cellulose. HA = hyaluronic acid. PVP =
polyvinylpyrrolidone. *= slurry autoclaved prior to addition of
gelling agent.
Methods of Preparation
[0029] Suspensions of the invention contain cyclosporin A Form 2
and a suspending agent. In another embodiment, the suspension also
contains one or more of water, buffer, and salt, in sufficient
quantities to provide a biocompatible formulation. By
"biocompatible," the inventors mean that the suspension is
appropriate for administration to the eye (for example, by
parenteral administration).
[0030] The formulations of the invention may be manufactured by
using either a heat-sterilized slurry of Form 2 cyclosporin mixed
aseptically with a sterile parenterally-biocompatible suspending
agent and other excipient; or by combining Form 2 cyclosporin with
a parenterally-biocompatible suspending agent and other excipients
and heat sterilizing the entire formulation.
[0031] These methods address various important problems with
cyclosporin formulation: 1) solid cyclosporin cannot be
pre-sterilized by irradiation without significant drug degradation
and formation of degradation products; 2) sterile filtration is
also not feasible because the formulation is a suspension; and 3)
terminal sterilization by heat will decrease gel viscosity. Also,
in one embodiment, the final viscosity of the drug formulation is
sufficiently high to keep the cyclosporin suspended throughout the
product's shelf-life. In another embodiment, the viscosity is
sufficiently low to permit the final formulation to flow through a
narrow gauge syringe, such as a 22, 23, 24, 25, or 26 gauge needle
or narrower. In still another embodiment, the formulation is
sufficiently high to keep the cyclosporin suspended throughout the
product's shelf-life, and also sufficiently low to permit the final
formulation to flow through a syringe with a 22, 23, 24, 25, or 26
gauge needle or narrower.
[0032] Methods 1 and 2, below, use hyaluronic acid as the
suspending agent but, other suitable suspending agents may be
substituted.
[0033] It should be noted that sterile hyaluronic acid is very
expensive and that method 2 provides a unique method of
sterilization, which allows the use of non-sterile hyaluronic acid
by heat-reducing the polymer to the correct molecular weight range,
so that it reaches the target viscosity range. Method 2, therefore,
requires precision manufacturing, where each new lot of hyaluronic
acid may shift to a different viscosity range, under identical
manufacturing conditions. Consequently, in order to assure the
correct viscosity range is reached in every commercial batch, the
heat cycle will need to be adaptive--that is--adjusted according to
a set of guidelines and experiments on the raw material lot prior
to manufacture of the drug product.
[0034] Furthermore, it should be noted that Method 2 prepares all
steps of the formulation in a single vessel. These two methods
allow for the rapid production of the drug product and
consequently, have substantial value in saving one day or more of
valuable manufacturing time over Method 1.
[0035] These methods depend on the inventors' surprising discovery
that cyclosporin A Form 2 may be autoclaved and still retain its
potency and stability. Other forms of cyclosporin--amorphous, Form
1 and Form 3--cannot be autoclaved, without unacceptable loss of
drug substance from the suspension.
Method 1--Aqueous Slurry Method
[0036] The appropriate amount of cyclosporin A Form 2 is suspended
and mixed in phosphate buffered saline solution and the slurry is
heat sterilized by autoclave. In an aseptic environment, the
appropriate amount of pre-sterilized hyaluronic acid is added to
the sterile cyclosporin slurry, is mixed, and then dissolved. The
drug product is brought to volume with sterile water for injection.
The final product has a viscosity in the correct range to create a
long-term stable suspension, while allowing the final formulation
to flow through a syringe fitted with a narrow-gauge needle, such
as 25 gauge needle or narrower.
Method 2--Single Vessel Method
[0037] An excess of non-sterile hyaluronic acid is dissolved in
phosphate buffered saline solution. Cyclosporin A Form 2 is
suspended and mixed. The resulting suspension formulation is
heat-sterilized by autoclave (using an "adaptive" heat cycle), at
the appropriate temperature and for the appropriate amount of time,
to both sterilize the formulation and bring the viscosity into the
desired range.
[0038] For parenteral formulations, it may be desirable to achieve
a viscosity that is sufficiently high to keep the cyclosporin
suspended throughout the product's shelf-life, and also
sufficiently low to permit the final formulation to flow through a
syringe with a 22, 23, 24, 25, or 26 gauge needle or narrower.
While hydrogel solutions are generally recognized as safe for
topical use, very few have been used for parenteral administration,
and none have been demonstrated to be safely injected through a 25
gauge needle (or narrower) into subconjunctival tissue at high
hydrogel concentrations. A high concentration of suspending agent
(up to 25%) is necessary in order to maintain the suspendability of
the 5-40% cyclosporin parenteral formulations described herein. In
one embodiment, parenteral formulations for use in subconjunctival
tissue are (1) injectable through a narrow-gauge needle, such as 25
gauge or narrower, in order to minimize tissue damage by the
needle, to allow for quick healing of the needle entry-point, and
to limit the back-flow of the injected formulation; (2) sterile;
(3) biocompatible; and (4) sufficiently viscous to maintain
suspendability throughout the shelf-life of the formulation and to
prevent tissue reflux out of the subconjunctival space. In such
formulations viscosity is sufficiently high to retain long-term
suspendability of the drug but sufficiently low to allow the entire
formulation to readily pass through a narrow gauge needle.
[0039] In one embodiment of the invention, the formulations have a
very high viscosity (e.g., .gtoreq.100,000 cps) yet may still able
to be injected out of syringe through a narrow-gauge needle. The
following table gives examples of such formulations.
TABLE-US-00005 Formulation 5% CsA, 3.5% HA 10% CsA, 3.5% HA 20%
CsA, 2.0% HA (10203X) (10204X) (10205X) Viscosity: TBD Viscosity:
1,300,000 cps Viscosity: 700,000 cps Needle size BD TSK Steriject
BD TSK Steriject BD TSKSteriject and type Precision 27G .times.
0.5'' Precision 27G .times. 0.5'' Precision 27G .times. 0.5'' Glide
UTW (Ultra Glide UTW (Ultra Glide UTW (Ultra 27G .times. 0.5'' Thin
Wall) 27G .times. 0.5'' Thin Wall) 27G .times. 0.5'' Thin Wall)
Needle Needle Needle Needle Needle Needle Injectabiltiy
Methods of Treatment
[0040] Compositions of the invention may be used to treat any
condition of the eye which is known to be amenable to topical
treatment with cyclosporin A (such as with Restasis.RTM.) at the
concentrations stated here. For example, compositions of the
invention may be used to treat patients suffering from dry eye, to
treat blepharitis and meibomian gland disease, to restore corneal
sensitivity that has been impaired due to refractive surgery on the
eye, to treat allergic conjunctivitis and atopic and vernal
keratoconjunctivitis, and to treat pterygium, conjunctival and
corneal inflammation, keratoconjunctivitis, graft versus host
disease, post-transplant glaucoma, corneal transplants, mycotic
keratitis, Thygeson's superficial punctate keratitis, uveitis, and
Theodore's superior limbic keratoconjunctivitis, among other
conditions.
[0041] The International Dry Eye Workshop (DEWS) defines dry eye as
"a multifactorial disease of the tears and ocular surface that
results in symptoms of discomfort, visual disturbance, and tear
film instability with potential damage to the ocular surface,
accompanied by increased osmolarity of the tear film and
inflammation of the ocular surface." It includes those conditions,
such as keratoconjunctivitis sicca, that are caused by tear
deficiency or excessive evaporation of tears.
[0042] Blepharitis is a chronic disorder producing inflammation of
the anterior and posterior lid margin, with involvement of skin and
its related structures (hairs and sebaceous glands), the
mucocutaneous junction, and the meibomian glands. It can also
affect the conjunctiva, tear film, and the corneal surface in
advanced stages and may be associated with dry eye. Blepharitis is
commonly classified into anterior or posterior blepharitis, with
anterior affecting the lash bearing region of the lids, and
posterior primarily affecting the meibomian gland orifices.
[0043] Meibomian gland disease most often occurs as one of three
forms: primary meibomitis, secondary meibomitis, and meibomian
seborrhea. Meibomian seborrhea is characterized by excessive
meibomian secretion in the absence of inflammation (hypersecretory
meibomian gland disease). Primary meibomitis, by contrast, is
distinguished by stagnant and inspissated meibomian secretions
(obstructive hypersecretory meibomian gland disease). Secondary
meibomitis represents a localized inflammatory response in which
the meibomian glands are secondarily inflamed in a spotty fashion
from an anterior lid margin blepharitis.
[0044] Impaired corneal sensitivity often occurs after refractive
surgery, such as photorefractive keratectomy, laser assisted
sub-epithelium keratomileusis (LASEK), EPI-LASEK, customized
transepithelial non-contact ablation, or other procedures in which
the corneal nerves are severed. Impaired corneal sensitivity may
also occur after viral infection, such as by HSV-1, HSV-2, and VZV
viruses. Patients with impaired corneal sensitivity often complain
that their eyes feel dry, even though tear production and
evaporation may be normal, suggesting that "dryness" in such
patients is actually a form of corneal neuropathy that results when
corneal nerves are severed by surgery or inflamed after viral
infection.
[0045] Allergic conjunctivitis is an inflammation of the
conjunctiva resulting from hypersensitivity to one or more
allergens. It may be acute, intermittent, or chronic. It occurs
seasonally, that is, at only certain time of the year, or it occurs
perennially, that is, chronically throughout the year. Symptoms of
seasonal and perennial allergic conjunctivitis include, in addition
to inflammation of the conjunctiva, lacrimation, tearing,
conjunctival vascular dilation, itching, papillary hyperplasia,
chemosis, eyelid edema, and discharge from the eye. The discharge
may form a crust over the eyes after a night's sleep.
[0046] Atopic keratoconjunctivitis is a chronic, severe form of
allergic conjunctivitis that often leads to visual impairment.
Symptoms include itching, burning, pain, redness, foreign body
sensation, light sensitivity and blurry vision. There is often a
discharge, especially on awakening from a night's sleep; the
discharge may be stringy, ropy, and mucoid. The lower conjunctiva
is often more prominently affected than the upper conjunctiva. The
conjunctiva may range from pale, edematous, and featureless to
having the characteristics of advanced disease, including papillary
hypertrophy, subepithelial fibrosis, fornix foreshortening,
trichiasis, entropion, and madarosis. In some patients the disease
progresses to punctate epithelial erosions, corneal
neovascularization, and other features of keratopathy which may
impair vision. There is typically goblet cell proliferation in the
conjunctiva, epithelial pseudotubular formation, and an increased
number of degranulating eosinophils and mast cells in the
epithelium. CD25+T lymphocytes, macrophages, and dendritic cells
(HLA-DR+, HLA-CD1+) are significantly elevated in the substantia
propria.
[0047] Like atopic keratoconjunctivitis, vernal
keratoconjunctivitis is a severe form of allergic conjunctivitis,
but it tends to affect the upper conjunctiva more prominently than
the lower. It occurs in two forms. In the palpebral form, square,
hard, flattened, closely packed papillae are present; in the bulbar
(limbal) form, the circumcorneal conjunctiva becomes hypertrophied
and grayish. Both forms are often accompanied by a mucoid
discharge. Corneal epithelium loss may occur, accompanied by pain
and photophobia, as may central corneal plaques and Trantas'
dots.
EXAMPLES
[0048] The invention is further illustrated by the following
examples.
[0049] When the inventors autoclaved aqueous suspensions of
cyclosporin A, the drug particles aggregated, making the product
unacceptable. Additionally, the inventors found that hyaluronic
acid also degrades upon autoclaving, causing a marked drop in
viscosity. Lower viscosity, in turn, reduces the suspendability of
the drug particles and causes them to settle. Formulations having
drug particles in suspension that too rapidly settle, or
irreversibly settle, may be useful for laboratory tests, but are
not commercially viable.
[0050] The inventors explored formulations of four cyclosporin A
polymorphic forms, the amorphous form, the tetragonal crystalline
form (form 1), the orthorhombic form (form 3), and cyclosporin A
Form 2.
[0051] A suspension of form 1 converts to the amorphous form and
aggregates upon autoclaving; clumping of the cyclosporin is also
observed. Consequently, neither form 1 nor the amorphous form is
suitable for autoclave stabilization. Furthermore, an autoclaved
suspension of F3 in water lost 11-28% of its potency during
autoclaving (Table 4); this, too, is unacceptable. In contrast, a
suspension of Form 2 in water was quite stable to autoclaving,
resisting degradation when compared to a pre-sterilization control.
X-ray analysis of filtered solid from the Form 2 formulation also
confirms that Form 2 is polymorphically stable to autoclaving (FIG.
3). These latter two findings are extremely surprising, considering
the lack of either chemical or polymorphic stability of the other
three forms.
[0052] The inventors explored the autoclavability of a series of
concentrated solutions of various polymers (no drug) which, when
loaded in a syringe, will flow through a narrow-gauge needle (25
gauge or narrower). The polymers evaluated were as follows:
cross-linked hyaluronic acid (Juvederm.RTM.), carbomer,
carboxymethylcellulose-medium molecular weight,
carboxymethylcellulose-high molecular weight,
hydroxyethylcellulose, hydroxypropylcellulose, Pluronic F127 and
polyvinylpyrrolidone K90. All of these are readily available from
commercial suppliers.
[0053] One hundred microliters of each of the autoclaved solutions
was injected into rabbit conjunctiva, in order to evaluate the
propensity for causing inflammation. Those polymers producing an
inflammatory reaction were eliminated from consideration (FIG. 4,
carbomer, both CMC's, and HPMC were eliminated). Additionally,
Juvederm.RTM. was eliminated because it formed a long-lasting bleb
which, in humans, might cause irritation as the eyelid moves over
the site of injection. Both HPMC and Pluronic separated from the
solution during/after autoclaving and consequently were also
eliminated. Of the commercially viable hydrogels, only HEC and PVP
demonstrated that they produced no inflammation in rabbit
conjunctiva after autoclaving. These two hydrogels were used to
formulate cyclosporin A suspensions for further evaluation. The
results of the studies are shown in Table 5.
[0054] Initially, the inventors explored the possibility of
heat-sterilizing a slurry of cyclosporin A of Form 1 (which
converts to the amorphous form). This approach resulted in
agglomeration of the drug and consequently, the formulation was not
viable. Further studies, adding PVP to suppress the agglomeration
of Form 1/amorphous form, also failed.
[0055] Since heat-sterilization of an aqueous suspension of
cyclosporin did not appear to be viable, the inventors planned to
prepare suspensions by aseptic technique, using pre-sterilize solid
cyclosporin. Various solid cyclosporins (Forms 1, 2, and 3 and
amorphous) were treated with gamma or e-beam irradiation. In all
cases, significant loss of drug (3-9%) occurred (FIG. 2 and Table
1). Furthermore, the substantial loss of drug indicates that high
levels of degradation products (around 3-9%) are generated in the
irradiation-sterilized material. These impurities may have negative
toxicological and/or regulatory implications; consequently, this
approach to sterilization appears to be undesirable.
TABLE-US-00006 TABLE 1 Effect of Irradiation Sterzation on
Cyclosporin (CsA) Drug Substance (solid) Sterzation Form 1 CsA Form
2 CsA Form 3 CsA Amorph. CsA Mode (Potency and Imp.) (Potency and
Imp.) (Potency and Imp.) (Potency and Imp.) None 98.4% w/w 94.6%
w/w 97.7% w/w 96.5% w/w Total Imp: 0.6% Total Imp: 0.6% Total Imp:
0.8% Total Imp: 0.7% 15 kGy Gamma 93.9% w/w 91.8% w/w 94.3% w/w
92.1% w/w % Rel. Change: % Rel. Change: 2.9% % Rel. Change 3.6% %
Rel. Change: 4.5% Total Imp: 1.8% Total Imp: 1.3% 4.6% Total Imp:
1.7% Total Imp: 1.4% 33 kGy Gamma 90.7% w/w 88.5% w/w 91.0% w/w
87.7% w/w % Rel. Change: % Rel. Change: 6.4% Rel. Change: 6.9% %
Rel. Change: 7.8% Total Imp: 2.4% Total Imp: 2.3% 9.2% Total Imp:
2.8% Total Imp: 2.3% E-Beam 92.6% w/w 90.3% w/w 93.4% w/w 92.0% w/w
% Rel. Change: % Rel. Change: 4.6% % Rel. Change: 4.5% % Rel.
Change: 5.9% Total Imp: 1.7% Total Imp: 1.6% 4.7% Total Imp: 1.5%
Total Imp: 1.3%
[0056] Subsequently, the inventors attempted to irradiate solid
cyclosporin (Forms 1, 2, and 3 and amorphous), under the best
conditions above, at cold temperatures. No significant improvement
was noted with any of the Forms of cyclosporin (Table 2).
TABLE-US-00007 TABLE 2 Effect of E-Beam Sterzation of Cyclosporins
Under Cold Conditions CsA Drug Substance Sample CsA Potency for CsA
Potency 15 kGy CsA Potency 30 kGy Gamma CsA Potency Treatment
Control Sample Gamma Treatment Treatment E-Beam 15 kGy Treatment
Dry Ice 99.2% w/w 96.7% w/w 93.8 w/w 93.8% w/w (% Rel. Change:
2.5%) (% Rel. Change: 5.4%) (% Rel. Change: 5.4%) Cold Pack 96.5%
w/w 93.0% w/w 92.1% w/w 93.2% w/w (% Rel. Change: 3.6%) (% Rel.
Change: 4.6%) (% Rel. Change: 3.4%)
[0057] After it became apparent that irradiation of solid
cyclosporins produced too much degradation, the inventors attempted
to irradiate an aqueous suspension of cyclosporin, using hyaluronic
acid as a suspending agent. This approach resulted in 4-10%
degradation of the drug within the formulation.
TABLE-US-00008 TABLE 3 Effect of Sterilization by Irradiation on
Aqueous Suspensions of Cyclosporin [CsA] using Hyaluronic Acid [HA]
as a Suspending Agent, at Various Temperatures CsA Potency for CsA
Potency Post- % Relative Change in Sterilization Treatment Control
Sample Sterilization Potency Cold Pack Control CsA 103.2% w/w Not
Applicable Not Applicable Hydrogel Sample CsA-HA Sample (Cold Pack
) 103.2% w/w 98.9% w/w 4.2% Treated with 15 kGy Gamma CsA-HA Sample
(Cold Pack) 103.2% w/w 92.3% w/w 10.6% Treated with 30 kGy Gamma
CsA-HA Sample (Cold Pack ) 103.2% w/w 92.8% w/w 10.1% Treated with
E-Beam (15 kGy)
[0058] Finally, the inventor turned their focus on steam
sterilization of slurries and full formulations of cyclosporins.
Slurries of Form 1 (which converts to amorphous) agglomerate during
heat-sterilization. Slurries of Form 3, while physically stable and
more chemically stable than Form 1, degraded significantly during
heat sterilization. But, to the inventors' surprise, slurries of
Form 2 were both physically and chemically stable (Tables 4 and
5).
TABLE-US-00009 TABLE 4 Heat-Sterilization of Slurries of
Cyclosporin (ScA) Form 2 (F-2) in water CsA-F2 Slurry CsA-F3 Slurry
% % Initial 96.86 101.41 120.degree. C. 15 min 96.88 88.61
108.degree. C. 60 min 106.69 71.72
TABLE-US-00010 TABLE 5 Physical Stabiltiy of Forms 2 and 3 Before
and After Heat Sterilization Formulation Material Spec. D90 D50 D10
Conditions A CsA-F2 Slurry 198.6313 116.8544 8.2711 Slurry control
for steam control sterilization study A, autoclaved CsA-F2
Autoclaved 186.4431 99.902 7.0518 Autoclaved at 120 C. for slurry
15 minutes A, autoclaved CsA-F2 Autoclaved 195.603 112.532 9.209
Autoclaved at 108 C. for slurry 60 minutes B CsA-F3 Slurry 110.8281
63.3348 7.1711 Slurry control for steam control sterilization study
B, autoclaved CsA-F3 Autoclaved 116.8761 67.523 12.1564 Autoclaved
at 120 C. for slurry 15 minutes B, autoclaved CsA-F3 Autoclaved
115.556 65.3309 10.5518 Autoclaved at 108 C. for slurry 60 minutes
% potency compared to CsA Formulation Material Conditions Form 2
standard A CsA-F2 Control 96.9 A CsA-F2 120.degree. C., 15 min 96.9
A CsA-F2 108.degree. C., 60 min 106.7 B CsA-F3 Control 101.4 B
CsA-F3 120.degree. C., 15 min 88.6 B CsA-F3 108.degree. C., 60 min
71.7
Ocular Congestion
[0059] Parenterally-biocompatible suspending agents were identified
by injecting sterile concentrated solutions into the
subconjunctival space and evaluating the toxicological response. An
injection of 100 ul of the following polymers in phosphate buffered
saline was administered subconjunctivally to New Zealand white
rabbits and observed for a period of seven days.
[0060] 2% Carbomer (Carbopol Ultrez 10NF, Lubrizol)
[0061] 8% Carboxymethyl Cellulose (low viscosity CMC, Lubrizol)
[0062] 6% Carboxymethyl Cellulose (high viscosity CMC,
Lubrizol)
[0063] 6% HEC (Ashland)
[0064] 6% HPMC (Dow Chemical)
[0065] Juvederm Ultra (Allergan, Inc)
[0066] Pluronic F127 (BASF)
[0067] Polyvinyl pyrrolidone (PVP K90, BASF)
TABLE-US-00011 tech Alternative type name source Lot# info vendor
CoA Grade vendor Grade 1 PVP PVP K30 Sigma_ BCBB78 Mw 40K Sigma_
yes BASF PHEUR/USP/ Aldrich 59 (PSO: Aldrich NF/JP 81420- 5% in
500G (or water, PSO pH 3.6) R14247) 2 PVP PVP K90 Sigma_ BCBB39 Mw
Sigma_ yes BASF PHEUR/USP/ Aldrich 54 360K Aldrich NF/JP 81440-
250G 3 PVP PVP 10 Sigma- 050M00 Mw 10K Sigma_ yes BASF PHEUR/USP/
Aldrich 39 Aldrich NF PVP10- 500G 4 HPMC Hypromellose PSO XB1401
Sigma Dow yes USP/PHEUR (tested to JP) PM# 2N11 H3785: Chemical
1018 4000 (R19424) cP, 2% in water 5 CMC Carboxy PSO 96413 CMC from
methyl R19716 Ashland/ cellulose Q Aqualon is sodium pending
NF/USP, 6 CMC Carboxy PSO 96077 methyl R19717 cellulose sodium 7
Hydroxyethyl Natrosol Kevin F0854 Type Ashland HEC from cellulose
(Type Warner 250- Ashland./ (HEC) 250-HHX HHX Aqualon is pharm)
pharm USP/EP, 8 Acrylate/C10- Carbopol Kevin EC742E acrylate
Lubrizol USP/NF 30 Alkyl ETD Warner K343 crosspolymer acrylate
2020NF (Viscosity, 47-77K cP 0.5% wt at pH 7.5) 9 Carbomer Carbopol
Kevin CC83RZ type A Lubrizol USP/NF Interpolymer Ultrez 10 NF
Warner G726 (Viscosity, polymer 45-65K cP 0.5% wt at pH 7.5) 10
Carbomer- Carbopol Kevin EC863C type C Lubrizol USP/PHEUR/
Homopolymer 980 NF Warner C625 (Viscosity, JPE polymer 40-60K cP
0.5% wt at pH 7.5) 1 PVP PVP K30 Sigma_ BCBB78 Mw 40K Sigma_ yes
BASF PHEUR/USP/ Aldrich 59 (PSO: Aldrich NF/JP 81420- 5% in 500G
(or water, PSO pH 3.6) R14247) 2 PVP PVP K90 Sigma_ BCBB39 Mw
Sigma_ yes BASF PHEUR/USP/ Aldrich 54 360K Aldrich NF/JP 81440-
250G 3 PVP PVP 10 Sigma- 050M00 Mw 10K Sigma_ yes BASF PHEUR/USP/
Aldrich 39 Aldrich NF PVP10- 500G 4 HPMC Hypromellose PSO XB1401
Sigma Dow yes USP/PHEUR (tested to JP) PM# 2N11 H3785: Chemical
1018 4000 (R19424) cP, 2% in water 5 CMC Carboxy PSO 96413 CMC from
methyl R19716 Ashland/ cellulose Q Aqualon is sodium pending
NF/USP, 6 CMC Carboxy PSO 96077 methyl R19717 cellulose sodium 7
Hydroxyethyl Natrosol Kevin F0854 Type Ashland HEC from cellulose
(Type Warner 250- Ashland./ (HEC) 250-HHX HHX Aqualon is pharm)
pharm USP/EP, 8 Acrylate/C10- Carbopol Kevin EC742E acrylate
Lubrizol USP/NF 30 Alkyl ETD Warner K343 crosspolymer acrylate
2020NF (Viscosity, 47-77K cP 0.5% wt at pH 7.5) 9 Carbomer Carbopol
Kevin CC83RZ type A Lubrizol USP/NF Interpolymer Ultrez 10 NF
Warner G726 (Viscosity, polymer 45-65K cP 0.5% wt at pH 7.5) 10
Carbomer- Carbopol Kevin EC863C type C Lubrizol USP/PHEUR/
Homopolymer 980 NF Warner C625 (Viscosity, JPE polymer 40-60K cP
0.5% wt at pH 7.5)
Gross ocular congestion was shown to resolve within 7 days for CMC,
HEC, HPMC, Pluronic and PVP. Ocular discharge was shown to resolve
within three days. Ocular discharge resolved within 3 days for all
groups except one. Results of the experiment are provided in FIGS.
9-11.
Impurity and Potency Analysis
[0068] The inventors prepared various formulations and evaluated
their potency and purity, as well particle size distribution.
TABLE-US-00012 Impurities Analysis Pre- Post- Composition Autoclave
Autoclave CsA CsA CsA Particle Potency (%) Total Total Absolute
Size CsA HEC No Impurities Impurities Change Formulation (.mu.m)
(%) (%) autoclave Autoclave (% a/a) (% a/a) (% a/a) HEC-1 10 5 5
117.20% 115.70% 0.71% 0.69% -0.02% HEC-2 10 20 5 103.60% 116.60%
0.61% 0.61% 0.00% HEC-3 10 5 2 116.40% 118.80% 0.78% 0.70% -0.08%
HEC-4 10 20 2 124.50% 124.70% 0.73% 0.69% -0.04% HEC-5 25 5 5
126.70% 116.60% 0.58% 0.58% 0.00% HEC-6 25 20 5 140.00% 147.40%
0.56% 0.56% 0.00% HEC-7 25 5 2 137.50% 142.50% 0.63% 0.59% -0.04%
HEC-8 25 20 2 129.50% 119.70% 0.56% 0.57% 0.01% HEC-9 10 10 3
118.60% 111.70% 0.61% 0.62% 0.01%
TABLE-US-00013 Composition CsA Particle Potency (%) Size CsA PVP90
No Formulation (.mu.m) (%) (%) autoclave Autoclave PVP-1 10 5 25
102.51 101.01 PVP-2 10 20 25 113.81 111.82 PVP-3 10 5 15 122.42
114.04 PVP-4 10 20 15 120.28 123.3 PVP-5 25 5 25 118.56 118.46
PVP-6 25 20 25 114.55 115.28 PVP-7 25 5 15 116.37 115.66 PVP-8 25
20 15 120.9 124.05 PVP-9 10 10 25 132.51 136.36 PVP-10 25 10 25
118.03 126.6
TABLE-US-00014 Autoclave Conditions CsA Temp (.degree. C.)/ Crystal
Time Particle size distribution Lot # Form Excipient (min.) D90 D50
D10 1 2 5% CMC None 52.38 10.80 5.31 2 2 5% CMC 121/10 18.02 11.55
5.74 3 3 3% CMC None 28.01 12.09 6.84 4 3 3% CMC 121/10 20.31 11.27
6.56 5 2 None None 198.63 116.85 8.27 6 2 None 120/15 186.44 99.90
7.05 7 2 None 108/60 195.60 112.53 9.21 8 3 None None 110.83 63.33
7.17 9 3 None 121/15 116.88 67.52 12.16 10 3 None 108/60 115.56
65.33 10.55 11 2 None None 13.15 9.12 6.17 12 2 None 121/15 14.15
9.12 6.42 13 2 None None 14.14 9.66 6.44 14 2 None 121/15 14.30
9.37 5.95
TABLE-US-00015 TABLE 5 Key F2 Formulation Properties of Evaluated
Polymers In-vivo Autoclavability tolerability (1 wk CSA-F2
Syringeability (121.degree. C., 15 min.) sub-conJ.) Setting Potency
Carbopol Max.cone. 4% No visible Poorly tolerated na na w/22 G
change (congestion) Carboxymethyl Cellulose Max.cone. 9% No visible
Poorly tolerated na na (CMC) medium viscosity w/22 G change
(congestion) Carboxymethyl Cellulose Max.cone. 6% No visible Poorly
tolerated na na (CMC) high viscosity w/22 G change (congestion)
Hydroxyethyl Cellulose Max.cone. 6% No visible Well tolerated. No
setting in No loss in (HEC) w/22 G change Slight congestion
comparison potency compared to with BDP gel post- saline under same
autoclave conditions Hydroxyethyl Methyl Max.cone. 7% Full
formation Well tolerated. na na Cellulose (HPMC) w/22 G visibly
separates with Comparable to moist heat sterilzation saline.
Juvaderm Ultra 30 g, as formulated Pre-sterilized by Poorly
tolerated na na by manufacturer manufacturer (swelling at 1 week)
Pluronic F127 Max.cone. 40% Placebo visibly Tolerated. Slight na na
w/22 G separates with moist congestion and discharge heat
sterilization compared to saline. Polyvinylpyrrolidone K90
Max.cone. 27% No visible Ok Some setting No loss in (PVPK90) w/22 G
change with syringeable potency concentrations post- (but
acceptable) autoclave
* * * * *