US006613358B2
(12) United States Patent
(10) Patent N0.:
(45) Date of Patent:
Randolph et al.
(54) SUSTAINED-RELEASE COMPOSITION
Brems et al., “Equilibrium Denaturation of Insulin and
Proinsulin,” Biochemistry 29: 9289—9293 (1990).
INCLUDING AMORPHOUS POLYMER
Ct., NiWot, CO (US) 80503; Mark C.
Casal et al., “Subunit Interface of Triosephosphate
Isomerase: Site—Directed Mutagenesis and Characterization
of the Altered Enzyme,” Biochemistry 26: 1258—1264
Manning, 1112 Live Oak Ct., Fort
(1987).
(76) Inventors: Theodore W. Randolph, 7916 Sussex
Collins, CO (US) 80525; Richard F.
Falk, 1266 NW. Knoxville, Apt. D.,
Bend, OR (US) 97701
(*)
Notice:
US 6,613,358 B2
Sep. 2, 2003
Subject to any disclaimer, the term of this
patent is extended or adjusted under 35
U.S.C. 154(b) by 55 days.
Cory et al., “Use of an Aqueous Soluble Tetrazolium/
Formazan Assay for Cell GroWth Assays in Culture,” Can
cer Commun., 3: 207—2112 (1991).
Davis et al., “Preparation and Characterization of Antibodies
With Speci?city for the Amino—Terminal Tetrapeptide
Sequence of the Platelet—Derived Connective Tissue Acti
vating Peptide—III,” Biochem. Intl., 10: 394—414 (1985).
(21) Appl. N0.: 09/877,330
Erikson et al., “Solid—Phase Peptide Synthesis,” in The
Proteins, 3rd ed., vol. 2, Neurath et al. (eds.), pp. 257—527
(22) Filed:
Jun. 7, 2001
(1976).
(65)
Prior Publication Data
Ettinger and Timasheff, “Optical Activity of Insulin. I. On
the Nature of the Circular Dichroism Bands,” 10: 824—83
(1971).
US 2002/0132007 A1 Sep. 19, 2002
Continuation of application No. 09/403,412, ?led as appli
Finn et al., “The Synthesis of Peptides by Solution Methods
and Emphasis on Peptide Hormones,” in The Proteins, 3rd
ed., vol. 2, Neurath et al. (eds.), pp. 105—253, Academic
Press, NeW York (1976).
cation No. PCT/US99/06198 on Mar. 18, 1999, now aban
doned.
Fraser et al., “Topoisomerase Ila Promoter Trans—Activa
tion Early in Monocytic Differentiation of HL—60 Human
(60)
Provisional application No. 60/166,230, ?led on Nov. 18,
1999, and provisional application No. 60/078,390, ?led on
(51)
(52)
Int. Cl.7 ............................ .. A61K 9/16; A61F 2/00
US. Cl. ..................... .. 424/489; 424/423; 424/426;
Leukemia Cells,” Mol. Pharmacol. 47:696—706 (1995).
Johnson, “Protein Secondary Structure and Circular Dichro
ism: A Practical Guide,” Genetics 7: 205—214 (1990).
Miyajima et al., “Effect of polymer crystallinity on papav
Related US. Application Data
(63)
Mar. 18, 1998.
424/427; 424/434; 424/435; 514/772.3;
514/781; 514/937; 514/951
(58)
Field of Search ............................... .. 424/423, 426,
424/427, 428, 434, 435, 464, 489
(56)
erine release from poly (1—lactic acid) matriX,”J. Controlled
Release 49: 207—215 (1997).
Melberg and Johnson, “Changes in Secondary Structure
FolloW With Dissociation of Human Insulin HeXamers: A
Circular Dichroism Study,” Genetics 8: 280—286 (1990).
Merri?eld, “Solid—Phase Peptide Synthesis,” in Chem.
References Cited
Polypeptides, Katsoyannis and Panayotis (eds.), pp.
335—361 (1973).
U.S. PATENT DOCUMENTS
Merri?eld, “Solid Phase Peptide Synthesis. I. The Synthesis
3,941,763 A
4,308,280 A
4,526,938 A
3/1976
Sarantakis ............. .. 260/1125
12/1981 Sportoletti et al. ....... .. 424/311
7/1985 Churchill et al. ......... .. 525/415
4,582,820
A
4/1986
Teng
. . . . . . . . . . . . . . . .
. . . . . ..
514/3
4,873,187
A
10/1989
Taub
. . . . . . . . . . . .
. . . . . ..
435/5
5,010,183
A
5,043,280 A
4/1991
Macfarlane
... ... .
. . . . . . ..
8/1991 Fischer et al.
536/27
435/235.1
5,364,884 A
5,445,832 A
11/1994 Varma et al. ............. .. 514/551
8/1995 Orsolini et al. ........... .. 424/491
5,770,559 A
6/1998 Manning et al. ............. .. 514/2
of a Tetrapeptide,” J. Am. Chem. Soc., 85: 2149—2154
(1963).
Mitsunobu, “The Use of Diethyl AzodicarboXylate and
Triphenylphosphine in Synthesis and Transformation of
Natural Products,” Synthesis, pp. 1—28 (1981).
Pocker and BisWas, “Conformational Dynamic of Insulin in
Solution. Circular Dichroic Studies,” Biochemistry 19:
5043—5049 (1980).
(List continued on neXt page.)
FOREIGN PATENT DOCUMENTS
WO
94/08599
4/1994
OTHER PUBLICATIONS
Ahern et al., “Control of oligomeric enzyme thermostability
by protein engineering,” Proc. Natl. Acad. Sci USA 84:
675—679 (1987).
ArakaWa and Timasheff, “Preferential Interactions of Pro
teins With Salts in Concentrated Solutions,” Biochemistry
21: 6545—6552 (1982).
ArakaWa and Timasheff, “Stabilization of Protein Structure
by Sugars,” Biochemistry 21: 6536—6544 (1982).
Baker et al., “The Structure of 2Zn Pig Insulin Crystals at 1.5
A Resolution,” Phil. Trans. R. Soc. London B319: 369—456
(1989).
Primary Examiner—James M. Spear
(74) Attorney, Agent, or Firm—Medlen & Carroll LLP
(57)
ABSTRACT
Provided is a sustained release composition for sustained
release of a pharmaceutical substance. The composition
includes a biocompatible polymer that is highly amorphous
and a pharmaceutical substance in a hydrophobic ion com
pleX With an amphiphilic material. Also provided is a
compressed antisolvent method for manufacturing the
composition, various product forms incorporating the com
position and various uses for the composition.
48 Claims, 35 Drawing Sheets
US 6,613,358 B2
Page 2
OTHER PUBLICATIONS
Sambrook et al., Molecular Cloning: A Laboratory Manual,
Matsuura et al., “Structure and Stability of Insulin Dissolved
in 1—Octanol,” J. Am. Chem. Soc., 115: 1261—1264 (1993).
Cold Spring Harbor Laboratory, (1989).
Meyer et al., “Generation of soluble and active subtilisin and
Stewart and Young, Solid Phase Peptide Synthesis, Freeman
ot—chymotrypsin in organic solvents via hydrophobic ion
pairing,” Int. J. Peptide Prot. Res., 47: 177—181 (1996).
and Company, San Francisco (1969) (Title and Copyright
Pages Only).
van Stokkum et al., “Estimation of Protein Secondary Struc
ture and Error Analysis from Circular Dichroism Spectra,”
Meyer et al., “Solution Behavior of ot—Chymotrypsin Dis
solved in Nonpolar Organic Solvents Via Hydrophobic Ion
Anal. Biochem, 191:110—118 (1990).
Pairing,” Biopolymers 35: 451—456 (1995).
WeitZel et al., “Insulinahnliche Aktivitat von Arginylverbin
dungen in vitro,” Hoppe Seylers Z. Physiol. Chem,
352:1617—1630 (1971).
Meyer et al., “Selective Precipitation of Interleukin—4 Using
Hydrophobic Ion Pairing: A Method for Improved Analysis
WlodaWer et al., “Structure of Bovine Pancreatic Trypsin
Inhibitor. Results of Joint Neutron and X—ray Re?nement of
Serum Albumin,” Pharm. Res., 11: 1492—1495 (1994).
Crystal Form II,” J. Mol. Biol., 180: 301—329 (1984).
WlodaWer et al., “Comparison of TWo Highly Re?ned
Structures of Bovine Pancreatic Trypsin Inhibitor,” J. Mol.
Biol, 193:145—156 (1987).
Stratton et al., “Drug Delivery Matrix Containing Native
protein Precipitates Suspended in a PoloXamer Gel,” J.
Pharm. Sci., 86: 1006—1010 (1997).
Falk et al., “Controlled release of ionic compounds from
poly (L—lactide) microspheres produced by precipitation
With a compressed antisolvent,” J. Controlled Release
44:77—85 (1997).
of Proteins Formulated With Large EXcesses of Human
PoWers et al., “Enhanced Solubility of Proteins and Peptides
in Nonpolar Solvents Through Hydrophobic Ion Pairing,”
Biopolymers 33: 927—932 (1993).
Randolph et al., “Sub—micrometer—siZed biodegradable par
ticles of poly (L—lactic acid) via the gas antisolvent spray
precipitation process,” Biotechnol. Prog., 9: 429—435
(1993).
Nielsen, Mechanical Properties of Polymers, (NeW York:
Reinhold Publishing Corporation) pp. 235—236 (1962).
U.S. Patent
Sep. 2, 2003
Sheet 1 0f35
US 6,613,358 B2
;uagamaoo uogmmd wamdde 8!“ J0 301
U.S. Patent
Sep. 2, 2003
Sheet 2 0f 35
S:o
US 6,613,358 B2
xmQ F2x
w
v
OwUcFmCZaoI
QQ X
0
a
LOGARITHM OF THE APPARENT PARTITKJN
COEFFICIENT
N.QE
U.S. Patent
US 6,613,358 B2
m.
x
.
m
0
2.,
0
w
L
o
M
m.@E
owcmomA:o
_
F
m
.
_
.
_
.
_
h
.
.
o
o
o
N
is
O.
‘H.
M;
mapggaoa uogmmd luamddc 9111 JD 30]
.E“53m2a:m
U.S. Patent
Sep. 2, 2003
Sheet 4 0f35
US 6,613,358 B2
c:
\O
("I
L.
0
..ln
(‘1
o
"'3'
f“
A
’
E
o
5
N
,5;
-
‘an
Q5
(.9
N
LL
_
;,
C1
ca
--v—4
0]
6
"C
N
C:
I
I
O
Q
C
o
O
c
C
Q
O
O
Q
o
I
O
a
Q
Q
c
0
o
o
Q
0
Menu Residue Elliplicity
-
U.S. Patent
Sep. 2, 2003
US 6,613,358 B2
Sheet 5 0f 35
\n
In
(\I
If)
~
wt
0!
A59352
m3m8m3
_
._
258.
._
._
.
oc om
oio w
082‘
2‘52.
Mean Residue Ellipticity
m.GE
U.S. Patent
Sep. 2, 2003
Sheet 7 0f35
US 6,613,358 B2
6
(‘Q
Q
0
" N
.2
H
N
M
N
-
H
2:
~
(2
m
m
I.
o
0
-
'
I
w-i
Q
,_,
.
.
,
P:
.
I
N
0
c
m
wapmaoa uomund waledde 30 lulmln?ol
2
m
U.S. Patent
Sep. 2, 2003
Sheet 8 0f35
US 6,613,358 B2
l ‘0000
0.8000
06000
ABSORNCE
0.4000
~
)/
\
/
0.20 00
0-0000
260.0
\\
270.0
280.0
290.0
WAVELENGTH IN nrn
FIG. 8
3(110 30.0
U.S. Patent
Sep. 2, 2003
Sheet 9 0f35
ca?
03?:
US 6,613,358 B2
,
_
O
5%
N
4:4:
0
Q§Q~
,
E
22
,
m
-
2..
2O
{a
_
H
..
C
2.
:50)'
2g
~L')
Q
m
I
120
Melting Point (° C)
O
U.S. Patent
l
MIEUDGRS
.L
:
SINCDRIGANWL
m
03O
Sep. 2, 2003
Sheet 10 0f 35
US 6,613,358 B2
I
I
I
l
("13
53
5
a»
l
l
so
815
9b 915 do 65
TEMPERATURE(°C)
1
!
no
H5
I
:20
FIG. 10
A255
>
E6 250
q 245
I
l
5240
l
9 235
_
L55230-
.
2225:
>
O
_
,
,
,
_
. , ,
,
.
. .
.
.
.
.
.
.
.
_
. ,
~
.
2
I
l
l
l
l
I
l
l
80
e5
90
95
I00
105
H0
H5
TEMPERATURE (°c)
FIG. 11
L
120
U.S. Patent
Sep. 2,2003
Sheet 11 0f35
102
19.4.
106
I
ANTISOLVENT
PRECIPITATION
116
108
11.9
__.__-----—>
SEPARATION
1 14
Y
112
FIG. 12
US 6,613,358 B2
U S Patent
Sep 2, 2003
US 6,613,358 B2
Sheet 12 0f 35
136
142
1Ki\l
W
134
132
126
FIG. 13
144
N\
130
U.S. Patent
Sep. 2, 2003
Sheet 13 0f 35
Fig. 14
US 6,613,358 B2
U.S. Patent
Sep. 2, 2003
Sheet 14 0f 35
US 6,613,358 B2
U.S. Patent
Sep. 2, 2003
'
Sheet 15 0f 35
US 6,613,358 B2
U.S. Patent
Sep. 2, 2003
Sheet 16 0f 35
US 6,613,358 B2
U.S. Patent
Sep. 2, 2003
Sheet 17 0f 35
US 6,613,358 B2
U.S. Patent
Sep. 2, 2003
Sheet 18 0f 35
132
FIG. 19
US 6,613,358 B2