U.S. patent application number 13/786377 was filed with the patent office on 2013-07-11 for method for formulating large diameter synthetic membrane vesicles.
This patent application is currently assigned to PACIRA PHARMACEUTICALS, INC.. The applicant listed for this patent is PACIRA PHARMACEUTICALS, INC.. Invention is credited to Kathleen D.A. Los, Ronald Warren McGuire, Ernest George Schutt, Peter Andrew Walters.
Application Number | 20130177634 13/786377 |
Document ID | / |
Family ID | 44761089 |
Filed Date | 2013-07-11 |
United States Patent
Application |
20130177634 |
Kind Code |
A1 |
Schutt; Ernest George ; et
al. |
July 11, 2013 |
METHOD FOR FORMULATING LARGE DIAMETER SYNTHETIC MEMBRANE
VESICLES
Abstract
The present invention generally relates to the field of
pharmaceutical sciences. More specifically, the present invention
includes apparatus and devices for the preparation of
pharmaceutical formulations containing large diameter synthetic
membrane vesicles, such as multivesicular liposomes, methods for
preparing such formulations, and the use of specific formulations
for therapeutic treatment of subjects in need thereof. Formation
and use of the pharmaceutical formulations containing large
diameter synthetic membrane vesicles produced by using the
apparatus and devices for therapeutic treatment of subjects in need
thereof is also contemplated.
Inventors: |
Schutt; Ernest George; (San
Diego, CA) ; McGuire; Ronald Warren; (Murrieta,
CA) ; Walters; Peter Andrew; (San Diego, CA) ;
Los; Kathleen D.A.; (San Diego, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
PACIRA PHARMACEUTICALS, INC.; |
San Diego |
CA |
US |
|
|
Assignee: |
PACIRA PHARMACEUTICALS,
INC.
San Diego
CA
|
Family ID: |
44761089 |
Appl. No.: |
13/786377 |
Filed: |
March 5, 2013 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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13083485 |
Apr 8, 2011 |
|
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13786377 |
|
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61322814 |
Apr 9, 2010 |
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Current U.S.
Class: |
424/450 ;
264/4.3; 264/4.6; 422/261; 422/307; 425/5; 514/330 |
Current CPC
Class: |
A61P 23/02 20180101;
B01J 13/125 20130101; A61K 9/127 20130101; A61K 31/445 20130101;
B01F 7/0075 20130101; A61K 9/1277 20130101; B01F 3/0807 20130101;
B01F 3/04049 20130101; A61K 9/1271 20130101; A61M 11/00 20130101;
A61K 31/4458 20130101; B01F 2015/061 20130101; B01F 5/104 20130101;
B01J 13/043 20130101; A61K 9/4833 20130101; A61P 23/00 20180101;
B01F 15/065 20130101; B01F 3/088 20130101; B01F 2215/0032 20130101;
B01D 1/16 20130101 |
Class at
Publication: |
424/450 ; 425/5;
264/4.6; 264/4.3; 514/330; 422/307; 422/261 |
International
Class: |
A61K 9/48 20060101
A61K009/48 |
Claims
1. An atomizing nozzle apparatus, comprising: a first fluid conduit
and a second fluid conduit each having at least one entrance
orifice and at least one exit orifice; a fluid contacting chamber
having a top comprising at least one entrance orifice and having a
bottom comprising at least one exit orifice and connecting to the
at least one exit orifice of the first fluid conduit; and a third
fluid conduit, wherein the third fluid conduit annularly surrounds
a portion of the fluid contacting chamber.
2. The atomizing nozzle of claim 1, wherein the fluid contacting
chamber connects to the at least one exit orifice of the second
fluid conduit.
3. The atomizing nozzle of claim 1, wherein the at least one exit
orifice of the fluid contacting chamber and the at least one exit
orifice of the third fluid conduit are flush.
4. The atomizing nozzle of claim 1, wherein the at least one exit
orifice of the fluid contacting chamber is recessed within the at
least one exit orifice of the third fluid conduit.
5. The atomizing nozzle of claim 1, wherein the at least one exit
orifice of the fluid contacting chamber extends beyond the at least
one exit orifice of the third fluid conduit.
6. The atomizing nozzle of claim 1, wherein the first fluid conduit
and the second fluid conduit are co-axial for a first portion of
the first fluid conduit length.
7. The atomizing nozzle of claim 1, wherein the second fluid
conduit annularly surrounds a second portion of the first fluid
conduit.
8. The atomizing nozzle of claim 1, wherein a diameter of the fluid
contacting chamber is larger than a diameter of the first fluid
conduit.
9. The atomizing nozzle of claim 1, wherein the fluid contacting
chamber conically narrows in diameter from the top of the fluid
contacting chamber to the bottom of the fluid contacting
chamber.
10. The atomizing nozzle of claim 1, further comprising wherein the
fluid contacting chamber conically narrows in diameter from a point
below the top of the fluid contacting chamber to the bottom of the
fluid contacting chamber.
11. A process for preparing droplets using the atomizing nozzle
apparatus of claim 1, comprising: applying a first component to the
first fluid conduit; applying a second liquid to the second fluid
conduit; and applying a gas to the third fluid conduit; wherein the
gas exiting the third fluid conduit exit orifice impinges the
liquid exiting the at least one exit orifice of the fluid
contacting chamber, providing atomized droplets, wherein the
droplets have an average diameter from about 100 nm to about 100
.mu.M.
12. The process of claim 11, wherein the first liquid is an
emulsion comprised of: a first aqueous phase; and a first organic
phase comprising a first organic solvent.
13. The process of claim 12, wherein the first organic solvent is
chloroform or methylene chloride.
14. The process of claim 12, wherein the first organic phase
further comprises at least one amphipathic lipid and at least one
neutral lipid.
15. The process of claim 14, wherein the at least one amphipathic
lipid is selected from the group consisting of
phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine.
16. The process of claim 14, wherein the at least one amphipathic
lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol.
17. The process of claim 14, wherein the at least one neutral lipid
is selected from the group consisting of glycerol esters, glycol
esters, tocopherol esters, sterol esters, hydrocarbons and
squalenes.
18. The process of claim 14, wherein the at least one neutral lipid
is selected from the group consisting of triolein, tripalmitolein,
trimyristolein, trilinolein, tributyrin, tricaprylin, tricaproin,
and tricaprin.
19. The process of claim 12, wherein the first aqueous phase
further comprises a therapeutic agent.
20. The process of claim 19, wherein the therapeutic agent is
bupivacaine.
21. The process of claim 11, wherein the second liquid applied to
the second fluid conduit is a second aqueous phase.
22. The process of claim 21, wherein the second aqueous phase
further comprises at least one from the group of dextrose and
lysine or mixture thereof.
23. The process of claim 11, wherein the droplet comprises a first
component core and a second aqueous phase shell.
24. The process of claim 11, wherein the gas is nitrogen.
25. The process of claim 11, wherein the droplets have an average
diameter from about 10 .mu.M to about 80 .mu.M.
26. The process of claim 11, wherein the droplets have an average
diameter from about 20 .mu.M to about 60 .mu.M.
27. The process of claim 11, wherein the droplets have an average
diameter from about 35 .mu.M to about 45 .mu.M.
28. An atomized droplet comprising: an emulsion core, wherein the
emulsion core comprises i) a first aqueous phase; and ii) a first
organic phase comprising a first organic solvent; and an aqueous
phase shell, wherein said atomized droplet is made by a process
comprising combining a first component, an aqueous phase, and a gas
using the atomizing nozzle apparatus of claim 1, said process
comprising: applying a first component to the first fluid conduit;
applying an aqueous phase to the second fluid conduit; and applying
a gas to the third fluid conduit; wherein the gas exiting the third
fluid conduit exit orifice impinges the liquid exiting the at least
one exit orifice of the fluid contacting chamber, providing
atomized droplets, wherein the droplets have an average diameter
from about 100 nm to about 100 .mu.M.
29. An atomizing nozzle apparatus, comprising: an first fluid
conduit, a second fluid conduit and a third fluid conduit each
having at least one entrance orifice and at least one exit orifice;
a first fluid contacting chamber having a top comprising at least
one entrance orifice and having a bottom comprising at least one
exit orifice and connecting to the at least one exit orifice of the
first fluid conduit, wherein the second fluid conduit annularly
surrounds a portion of the first fluid contacting chamber; a second
fluid contacting chamber having a top comprising at least one
entrance orifice and having a bottom comprising at least one exit
orifice and connecting to the at least one exit orifice of the
first fluid contacting chamber, wherein the third fluid conduit
annularly surrounds a portion of the second fluid contacting
chamber; and a fourth fluid conduit, wherein the fourth fluid
conduit annularly surrounds a portion of the second fluid
contacting chamber.
30. The atomizing nozzle of claim 29, wherein the first fluid
contacting chamber connects to the at least one exit orifice of the
second fluid conduit.
31. The atomizing nozzle of claim 29, wherein the at least one exit
orifice of the second fluid contacting chamber and the at least one
exit orifice of the fourth fluid conduit are flush.
32. The atomizing nozzle of claim 29, wherein the at least one exit
orifice of the second fluid contacting chamber is recessed within
the at least one exit orifice of the fourth fluid conduit.
33. The atomizing nozzle of claim 29, wherein the at least one exit
orifice of the second fluid contacting chamber extends beyond the
at least one exit orifice of the fourth fluid conduit.
34. The atomizing nozzle of claim 29, wherein the first fluid
conduit and the second fluid conduit are co-axial for a first
portion of the first fluid conduits length.
35. The atomizing nozzle of claim 29, wherein the second fluid
conduit annularly surrounds a second portion of the first fluid
conduit.
36. The atomizing nozzle of claim 29, wherein a diameter of the
fluid contacting chamber is larger than a diameter of the first
fluid conduit.
37. The atomizing nozzle of claim 29, wherein the first fluid
contacting chamber conically narrows in diameter from the top of
the first fluid contacting chamber to the bottom of the first fluid
contacting chamber.
38. The atomizing nozzle of claim 29, wherein the second fluid
conduit and the third fluid conduit are co-axial for a first
portion of the second fluid conduits length.
39. The atomizing nozzle of claim 29, wherein the third fluid
conduit annularly surrounds a second portion of the second fluid
conduit.
40. The atomizing nozzle of claim 29, wherein a diameter of the
second fluid contacting chamber is larger than a diameter of the
exit orifice of the second fluid contacting chamber.
41. The atomizing nozzle of claim 29, wherein the second fluid
contacting chamber conically narrows in diameter from the top of
the second fluid contacting chamber to the bottom of the second
fluid contacting chamber.
42. A process for preparing droplets using the atomizing nozzle
apparatus of claim 29, comprising: applying a first liquid to the
first fluid conduit; applying a second liquid to the second fluid
conduit; applying a third liquid to the third fluid conduit; and
applying a gas to the fourth fluid conduit, wherein the gas exiting
the fourth fluid conduit exit orifice impinges the liquid exiting
the at least one exit orifice of the second fluid contacting
chamber, providing atomized droplets, wherein the droplets have an
average diameter from about 100 nm to about 100 .mu.M.
43. The process of claim 42, wherein: the first liquid is an
emulsion comprised of: i) a first aqueous phase; and ii) a first
organic phase comprising a first organic solvent; the second liquid
is a second organic phase; the third liquid is a second aqueous
phase.
44. The process of claim 42, wherein: the first liquid is an
emulsion comprised of: i) a first aqueous phase; and ii) a first
organic phase comprising a first organic solvent; the second liquid
is a second aqueous phase; the third liquid is a second organic
phase.
45. The process of claim 43, wherein the first organic solvent is
chloroform or methylene chloride.
46. The process of claim 43, wherein the first organic phase
further comprises at least one amphipathic lipid and at least one
neutral lipid.
47. An evaporation apparatus, comprising at least one atomizing
nozzle apparatus of claim 1 and means for evaporating an organic
solvent.
48. An evaporation apparatus, comprising: a solvent removal vessel
having a top, a bottom and a circular wall; at least one atomizing
nozzle; a carrier gas entrance orifice; a solvent removal gas exit
orifice centrally connected to the top; and a product exit orifice
connected to the bottom of the vessel.
49. The apparatus of claim 48, wherein the apparatus further
comprises at least two carrier gas entrance offices.
50. The apparatus of claim 48, wherein at least part of the solvent
removal vessel is jacketed.
51. The apparatus of claim 48, wherein the atomizing nozzle is
mounted to and extending through the top of the solvent removal
vessel.
52. The apparatus of claim 48, wherein the top of the solvent
removal vessel comprises a lid.
53. The apparatus of claim 48, further comprising a rinse nozzle
mounted to and extending through the top of the solvent removal
vessel.
54. The apparatus of claim 48, wherein the circular wall has a
central axis and the solvent removal gas exit orifice further
comprises a tube extending into the solvent removal vessel residing
along the central axis.
55. The apparatus of claim 54, wherein the atomizing nozzle is
angled at least 5 degrees measured off the central axis of the wall
and in a plane parallel to the wall nearest to it.
56. The apparatus of claim 55, wherein the carrier gas entrance
orifice is combined with the atomizing nozzle.
57. The apparatus of claim 48, wherein the solvent removal gas exit
orifice further comprises a tube extending into the solvent removal
vessel, wherein the tube is fitted with a narrowing cone.
58. The apparatus of claim 48, wherein the solvent removal gas exit
orifice further comprises a tube extending into the solvent removal
vessel, wherein the tube is fitted with a narrowing cone and an
annular ring.
59. The apparatus of claim 57, wherein the tube extends from about
1/3 to about 4/5 of the way into the solvent removal vessel.
60. The apparatus of claim 57, wherein the tube extends about 2/3
of the way into the solvent removal vessel.
61. The apparatus of claim 57, wherein a bottom tip of the
narrowing cone diameter is from about 1/1000 to about 1/5 of the
diameter of the inside of the solvent removal vessel.
62. The apparatus of claim 48, wherein the solvent removal gas exit
orifice diameter is less than 1/10 of a diameter of the inside of
the solvent removal vessel.
63. The apparatus of claim 48, wherein the at least one atomizing
nozzle is the atomizing nozzle apparatus of claim 1.
64. The apparatus of claim 48, wherein the at least one atomizing
nozzle is the atomizing nozzle apparatus of claim 29.
65. The apparatus of claim 48, wherein the ratio of the inside
diameter of the solvent removal vessel to the diameter of the
narrowing cone of the solvent removal gas exit orifice is between
approximately 5:1 and 100:1.
66. The apparatus of claim 48, wherein the ratio of the inside
diameter of the solvent removal vessel to the diameter of the
narrowing cone of the solvent removal gas exit orifice is between
approximately 20:1 and 60:1.
67. A process for preparing large diameter synthetic membrane
vesicles using the evaporation apparatus of claim 48, comprising:
introducing large diameter synthetic membrane vesicles pre-droplets
to the solvent removal vessel, wherein the large diameter synthetic
membrane vesicles pre-droplets comprise a first component core and
an aqueous phase shell; applying a carrier gas in a tangental
direction to the circular wall through the carrier gas entrance
orifice; and removing a solvent removal gas through the solvent
removal gas exit orifice to provide the large diameter synthetic
membrane vesicles suspension.
68. The process of claim 67, wherein the first component core
comprises a first aqueous phase and a first organic phase.
69. The process of claim 68, wherein the first organic phase
comprises a continuous first organic solvent.
70. The process of claim 69, wherein the first organic solvent is
chloroform or methylene chloride.
71. The process of claim 68, wherein the first organic phase
further comprises at least one amphipathic lipid and at least one
neutral lipid.
72. The process of claim 67, wherein the first component core is
first aqueous phase droplets as a suspension in a first organic
phase.
73. The process of claim 67, wherein the first aqueous phase
droplets have an average diameter of from about 10 nm to about 10
.mu.m, about 100 nm to about 5 .mu.m, or about 500 nm to about 2
.mu.m.
74. The process of claim 69, wherein the first aqueous phase
droplets have an average diameter of about 1 .mu.m.
75. The process of claim 67, wherein the carrier gas comprises
nitrogen.
76. The process of claim 69, wherein the solvent removal gas
comprises nitrogen and organic solvent.
77. The process of claim 67, wherein the carrier gas and the
solvent removal gas travel in a vortex in the solvent removal
vessel.
78. The process of claim 67, wherein the large diameter synthetic
membrane vesicles are multivesicular liposomes having a structure
including multiple non-concentric chambers and comprising at least
one amphipathic lipid and at least one neutral lipid.
79. The process of claim 78, wherein the at least one amphipathic
lipid is selected from the group consisting of
phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine.
80. The process of claim 78, wherein the at least one amphipathic
lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol, and
1,2-dioleoyl-sn-glycero-3-phosphoglycerol.
81. The process of claim 78, wherein the at least one neutral lipid
is selected from the group consisting of glycerol esters, glycol
esters, tocopherol esters, sterol esters, hydrocarbons and
squalenes.
82. The process of claim 78, wherein the at least one neutral lipid
is selected from the group consisting of triolein, tripalmitolein,
trimyristolein, trilinolein, tributyrin, tricaprylin, tricaproin,
and tricaprin.
83. The process of claim 78, wherein the multivesicular liposomes
further comprises a therapeutic agent.
84. The process of claim 83, wherein the therapeutic agent is
bupivacaine.
85. An evaporation apparatus, comprising at least one atomizing
nozzle apparatus of claim 1 and means for removing an organic
solvent from a droplet.
86. A process for preparing large diameter synthetic membrane
vesicles using the evaporation apparatus of claim 48, comprising:
introducing large diameter synthetic membrane vesicles pre-droplets
to the solvent removal vessel, wherein the large diameter synthetic
membrane vesicles pre-droplets comprise a first component core and
an aqueous phase shell; applying a carrier gas in a tangental
direction to the circular wall through the carrier gas entrance
orifice; removing a solvent removal gas through the solvent removal
gas exit orifice to provide pre-temperature treatment large
diameter synthetic membrane vesicles; introducing the
pre-temperature treatment large diameter synthetic membrane
vesicles to an outlet line; contacting the pre-temperature
treatment large diameter synthetic membrane vesicles with a hot
solution in the outlet line, wherein the hot solution has a
temperature ranging from about 30.degree. C. to about 100.degree.
C. to provide post-temperature treatment large diameter synthetic
membrane vesicles; transferring the post-temperature treatment
large diameter synthetic membrane vesicles to a continuous-flow
particle-concentration system or continuous phase exchange system;
cooling the post-temperature treatment large diameter synthetic
membrane vesicles to a second temperature to provide the large
diameter synthetic membrane vesicles; and isolating the large
diameter synthetic membrane vesicles suspension.
87. The process of claim 86, wherein the first component core
comprises a first aqueous phase and a first organic phase.
88. The process of claim 87, wherein the first organic phase
comprises a continuous first organic solvent.
89. The process of claim 88, wherein the first organic solvent is
chloroform or methylene chloride.
90. The process of claim 88, wherein the first organic phase
further comprises at least one amphipathic lipid and at least one
neutral lipid.
91. The process of claim 86, wherein the first component core is a
suspension of first aqueous phase droplets in a first organic
phase.
92. The process of claim 86, wherein the first aqueous phase
droplets have an average diameter of from about 10 nm to about 10
.mu.m, about 100 nm to about 5 .mu.m, or about 500 nm to about 2
.mu.m.
93. The process of claim 92, wherein the first aqueous phase
droplets have an average diameter of about 1 .mu.m.
94. The process of claim 86, wherein the carrier gas comprises
nitrogen.
95. The process of claim 88, wherein the solvent removal gas
comprises nitrogen and organic solvent.
96. The process of claim 86, wherein the carrier gas comprises
nitrogen and water vapor.
97. The process of claim 86, wherein the carrier gas and the
solvent removal gas travel in a vortex in the solvent removal
vessel.
98. The process of claim 86, wherein the large diameter synthetic
membrane vesicles are multivesicular liposomes having a structure
including multiple non-concentric chambers and comprising at least
one amphipathic lipid and at least one neutral lipid.
99. The process of claim 98, wherein the at least one amphipathic
lipid is selected from the group consisting of
phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine.
100. The process of claim 98, wherein the at least one amphipathic
lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol.
101. The process of claim 98, wherein the at least one neutral
lipid is selected from the group consisting of glycerol esters,
glycol esters, tocopherol esters, sterol esters, hydrocarbons and
squalenes.
102. The process of claim 98, wherein the at least one neutral
lipid is selected from the group consisting of triolein,
tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin.
103. The process of claim 98 wherein the multivesicular liposomes
further comprises a therapeutic agent.
104. The process of claim 103, wherein the therapeutic agent is
bupivacaine.
105. A composition comprising multivesicular liposomes having a
structure including multiple non-concentric chambers and comprising
at least one amphipathic lipid and at least one neutral lipid,
wherein said multivesicular liposomes are made by a process
comprising removing organic solvent from multivesicular liposomes
pre-droplets using the evaporation apparatus of claim 48, said
process comprising: introducing multivesicular liposomes
pre-droplets to the solvent removal vessel, wherein the large
diameter synthetic membrane vesicles pre-droplets comprise a first
component core and an aqueous phase shell; applying a carrier gas
in a tangental direction to the circular wall through the carrier
gas entrance orifice; and removing a solvent removal gas through
the solvent removal gas exit orifice to provide the large diameter
synthetic membrane vesicles.
106. A composition comprising large diameter synthetic membrane
vesicles made by the process of claim 67 or claim 86.
107. The composition of claim 106, wherein the large diameter
synthetic membrane vesicles are multivesicular liposomes having a
structure including multiple non-concentric chambers and comprising
at least one amphipathic lipid and at least one neutral lipid.
108. The composition of claim 107, wherein the at least one
amphipathic lipid is selected from the group consisting of
phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine.
109. The composition of claim 107, wherein the at least one
amphipathic lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phospho glycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol.
110. The composition of claim 107, wherein the at least one neutral
lipid is selected from the group consisting of glycerol esters,
glycol esters, tocopherol esters, sterol esters, hydrocarbons and
squalenes.
111. The composition of claim 107, wherein the at least one neutral
lipid is selected from the group consisting of triolein,
tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin.
112. The composition of claim 107 wherein the multivesicular
liposomes further comprises a therapeutic agent.
113. The composition of claim 112, wherein the therapeutic agent is
bupivacaine.
114. A continuous-flow emulsification system, comprising: a mixer,
comprised of a rotor and a stator; a recirculation loop, comprised
of one or more recirculation lines; a heat exchanger; one or more
outlet lines; one or more continuous phase inlet lines; and a
discontinuous phase inlet line; wherein the heat exchanger and the
mixer are connected together in the recirculation loop by one or
more recirculation lines; further wherein the one or more outlet
lines and one or more continuous phase inlet lines are connected to
the recirculation loop; further wherein the end of the
discontinuous phase inlet line is located within approximately
1/3.sup.rd of a rotor diameter from the rotor and approximately
1/3.sup.rd of a rotor diameter of the rotation axis of the rotor
and is in fluid communication with the rotor.
115. The emulsification system of claim 114, wherein a continuous
phase entrance line is connected to the recirculation loop upstream
of the mixer and downstream of the heat exchanger and an outlet
line is connected to the recirculation loop downstream of the mixer
and upstream of the heat exchanger.
116. The emulsification system of claim 114, further comprising an
emulsion, which emulsion recirculates through the recirculation
line back to the mixer an average of at least 5 or more times.
117. The emulsification system of claim 114, further comprising
emulsion droplets produced in the mixer, which are on average less
than 10 microns in diameter.
118. A continuous processing system, comprising: one or more
concentrator units, each unit comprising: a retentate vessel; a
particle suspension inlet line, connected to the retentate vessel;
a first outlet line, connecting the retentate vessel and a particle
concentrating device; a pump located along the first outlet line
between the retentate vessel and the particle concentrating device;
a second outlet line, leading to another concentrator unit or the
final product collection vessel; and means for removing or
exchanging solvent.
119. The system of claim 118, wherein the means for removing or
exchanging solvent are each independently selected from the group
consisting of a tangential flow filtration unit, a hydro-cyclone
unit, and a centrifugal separator.
120. The system of claim 118, further comprising a new suspending
medium inlet line connected to the retentate vessel.
121. The system of claim 118, wherein the means for removing or
exchanging solvent are each a tangential flow filtration unit.
122. The system of claim 118, wherein the means for removing or
exchanging solvent are each a centrifugal separator.
123. The system of claim 118, wherein the system comprises at least
one tangential flow filtration unit and at least one centrifugal
separator.
124. The system of claim 118, wherein the continuous processing
system is a continuous-flow particle-concentration system or
continuous phase exchange system.
125. A process for making multivesicular liposomes using the
atomizing nozzle apparatus of claim 1, comprising: applying a first
liquid to the first fluid conduit, wherein the first liquid
comprises an organic solvent; applying a second liquid to the
second fluid conduit; applying a pressurized gas to the third fluid
conduit to provide atomized droplets, wherein the pressurized gas
exiting the third fluid conduit exit orifice impinges the liquid
exiting the fluid contacting chamber exit orifice; and removing the
organic solvent from the atomized droplets, wherein less than 4000
ppm of the organic solvent remains in the atomized droplets.
126. The process of claim 125, wherein the first liquid is an
emulsion comprised of: a discontinuous aqueous phase; and a
continuous organic phase comprising the organic solvent.
127. The process of claim 126, wherein the organic solvent is
methylene chloride.
128. The process of claim 126, wherein the discontinuous aqueous
phase further comprises a therapeutic agent.
129. The process of claim 126, wherein the continuous organic phase
further comprises a therapeutic agent.
130. The process of claim 127 or 128, wherein the therapeutic agent
is bupivacaine or a salt thereof.
131. The process of claim 125, wherein the second liquid applied to
the second fluid conduit is an aqueous solution.
132. The process of claim 131, wherein the aqueous solution further
comprises dextrose and lysine.
133. The process of claim 125, wherein the gas is a sterilized
gas.
134. The process of claim 133, wherein the gas is nitrogen.
135. The process of claim 125 further comprising: introducing
atomized droplets to the evaporation apparatus of claim 48;
introducing a pressurized carrier gas tangentially to the circular
wall into the solvent removal vessel through the carrier gas
entrance orifice; and removing a solvent removal gas wherein the
solvent removal gas removes greater than 90% of the organic solvent
in the atomized droplets resulting in formation of multivesicular
liposomes.
136. The process of claim 135, wherein the carrier gas is heated
and humidified.
137. The process of claim 135, further comprising spraying a wall
rinse solution into the solvent removal vessel using a rinse
nozzle, wherein the wall rinse solution prevents build-up of
particles on the top of the evaporation apparatus.
138. The process of claim 125, wherein the atomized droplets
contain organic solvent in the range of from about 400 ppm to about
3500 ppm.
139. A process for making an emulsion using the emulsification
system of claim 114, comprising feeding an organic discontinuous
phase into the emulsification system through the discontinuous
phase inlet line and feeding an aqueous continuous phase into the
emulsification system through one or more continuous phase inlet
lines.
140. A process for making an emulsion using the emulsification
system of claim 114, comprising feeding an aqueous discontinuous
phase into the emulsification system through the discontinuous
phase inlet line and feeding an organic continuous phase fed into
the emulsification system through the one or more continuous phase
inlet lines.
141. The process of claim 140, wherein the organic continuous phase
is comprised of an organic solvent and a neutral lipid.
142. The process of claim 141, wherein the organic solvent is
methylene chloride.
143. The process of claim 140 wherein the aqueous discontinuous
phase is comprised of an acid and a therapeutic agent.
144. The process of claim 143, wherein the acid is phosphoric
acid.
145. The process of claim 143, wherein the therapeutic agent is
bupivacaine.
146. The process of claim 140, wherein a portion of the emulsion is
fed through one or more outlet lines to the inner fluid conduit of
the atomizing nozzle of claim 1.
147. The process of claim 140, wherein a portion of the emulsion is
fed through one or more outlet lines to the evaporation apparatus
of claim 48.
148. A process for preparing large diameter synthetic membrane
vesicles using an evaporation apparatus as disclosed and described
herein.
149. A large diameter synthetic membrane vesicles made by a process
as disclosed and described herein.
150. A composition comprising multivesicular liposomes comprising
an outer surface layer whose composition is different than the
composition of the internal structure.
Description
INCORPORATION BY REFERENCE TO ANY PRIORITY APPLICATIONS
[0001] Any and all priority claims identified in the application
Data Sheet, or any correction thereto, are hereby incorporated by
reference under 37 CFR 1.57.
FIELD OF THE INVENTION
[0002] The present invention generally relates to the field of
pharmaceutical sciences. More specifically, the present invention
relates to pharmaceutical formulations containing large diameter
synthetic membrane vesicles, such as multivesicular liposomes
(MVL), methods for preparing such formulations, and the use of
specific formulations for therapeutic treatment of subjects in need
thereof.
BACKGROUND
[0003] The following includes information that may be useful in
understanding the present embodiments. It is not an admission that
any of the information provided herein is prior art, or relevant,
to the presently described or claimed embodiments, or that any
publication or document that is specifically or implicitly
referenced is prior art.
[0004] Large scale methods of manufacturing large diameter
synthetic membrane vesicles, such as multivesicular liposomes,
often require large amounts of solvents, time sensitive steps and
concentration adjustment of the final product under sterile
conditions. In addition, current methods of manufacturing large
diameter synthetic membrane vesicles, such as multivesicular
liposomes on a commercial scale, require significant commitments in
manufacturing space, cost, and time. As such, developing stable
multivesicular liposome formulations containing a therapeutic agent
in a cost effective and timely manner remains an ongoing
challenge.
SUMMARY
[0005] Some embodiments provide an atomizing nozzle apparatus,
comprising a first fluid conduit and a second fluid conduit each
having at least one entrance orifice and at least one exit orifice,
a fluid contacting chamber having a top comprising at least one
entrance orifice and having a bottom comprising at least one exit
orifice and connecting to the at least one exit orifice of the
first fluid conduit, a third liquid channel, wherein the third
fluid conduit annularly surrounds a portion of the fluid contacting
chamber. In some embodiments, the fluid contacting chamber connects
to the at least one exit orifice of the second fluid conduit. In
some embodiments, the at least one exit orifice of the fluid
contacting chamber and the at least one exit orifice of the third
fluid conduit are flush. In some embodiments, the at least one exit
orifice of the fluid contacting chamber is recessed within the at
least one exit orifice of the third fluid conduit. In some
embodiments, the at least one exit orifice of the fluid contacting
chamber extends beyond the at least one exit orifice of the third
fluid conduit. In some embodiments, the first fluid conduit and the
second fluid conduit are co-axial for a first portion of the first
fluid conduit length. In some embodiments, the second fluid conduit
annularly surrounds a second portion of the first fluid conduit. In
some embodiments, a diameter of the fluid contacting chamber is
larger than a diameter of the first fluid conduit. In some
embodiments, the fluid contacting chamber conically narrows in
diameter from the top of the fluid contacting chamber to the exit
orfice of the fluid contacting chamber. In some embodiments, the
fluid contacting chamber conically narrows in diameter from a point
below the top of the fluid contacting chamber to the exit orfice of
the fluid contacting chamber.
[0006] Some embodiments provide a process for preparing droplets
using an atomizing nozzle as disclosed herein comprising applying a
first liquid to the first fluid conduit, applying a second liquid
to the second fluid conduit, applying a gas to the third fluid
conduit, wherein the gas exiting the third fluid conduit exit
orifice impinges the liquid exiting the at least one exit orifice
of the fluid contacting chamber, providing atomized droplets,
wherein the droplets have an average diameter from about 100 nm to
about 300 .mu.M. In some embodiments, the first liquid is an
emulsion comprised of a first aqueous phase, and a first organic
phase comprising a first organic solvent. In some embodiments, the
first organic solvent is chloroform or methylene chloride. In some
embodiments, the first organic phase further comprises at least one
amphipathic lipid and at least one neutral lipid. In some
embodiments, the at least one amphipathic lipid is selected from
the group consisting of phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine. In some embodiments, the at least one
amphipathic lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol. In some embodiments, the
at least one neutral lipid is selected from the group consisting of
glycerol esters, glycol esters, tocopherol esters, sterol esters,
hydrocarbons and squalenes. In some embodiments, the at least one
neutral lipid is selected from the group consisting of triolein,
tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin. In some embodiments, the
first aqueous phase further comprises a therapeutic agent. In some
embodiments, the therapeutic agent is bupivacaine. In some
embodiments, the second liquid applied to the second fluid conduit
is a second aqueous phase. In some embodiments, the droplet
comprises a first component core and a second aqueous phase shell.
In some embodiments, the gas is nitrogen. In some embodiments, the
droplets have an average diameter from about 20 .mu.M to about 60
.mu.M. In some embodiments, the droplets have an average diameter
from about 35 .mu.M to about 45 .mu.M.
[0007] Some embodiments provide atomized droplet comprising an
emulsion core, wherein the emulsion core comprises i) a first
aqueous phase; and ii) a first organic phase comprising a first
organic solvent; and an aqueous phase shell, wherein said atomized
droplet is made by a process comprising combining a first
component, an aqueous phase, and a gas using an atomizing nozzle as
disclosed and described herein, said process comprising applying a
first component to the first fluid conduit, applying an aqueous
phase to the second fluid conduit, and applying a gas to the third
fluid conduit, wherein the gas exiting the third fluid conduit exit
orifice impinges the liquid exiting the at least one exit orifice
of the fluid contacting chamber, providing atomized droplets,
wherein the droplets have an average diameter from about 100 nm to
about 300 .mu.M.
[0008] Some embodiments provide atomizing nozzle apparatus,
comprising an first fluid conduit, a second fluid conduit and a
third fluid conduit each having at least one entrance orifice and
at least one exit orifice, a first fluid contacting chamber having
a top comprising at least one entrance orifice and having a bottom
comprising at least one exit orifice and connecting to the at least
one exit orifice of the first fluid conduit, wherein the second
fluid conduit annularly surrounds a portion of the first fluid
contacting chamber, a second fluid contacting chamber having a top
comprising at least one entrance orifice and having a bottom
comprising at least one exit orifice and connecting to the at least
one exit orifice of the first fluid contacting chamber, wherein the
third fluid conduit annularly surrounds a portion of the second
fluid contacting chamber, and a fourth fluid conduit, wherein the
fourth fluid conduit annularly surrounds a portion of the second
fluid contacting chamber. In some embodiments, the first fluid
contacting chamber connects to the at least one exit orifice of the
second fluid conduit. In some embodiments, the at least one exit
orifice of the second fluid contacting chamber and the at least one
exit orifice of the fourth fluid conduit are flush. In some
embodiments, the at least one exit orifice of the second fluid
contacting chamber is recessed within the at least one exit orifice
of the fourth fluid conduit. In some embodiments, the at least one
exit orifice of the second fluid contacting chamber extends beyond
the at least one exit orifice of the fourth fluid conduit. In some
embodiments, the first fluid conduit and the second fluid conduit
are co-axial for a first portion of the first fluid conduits
length. In some embodiments, the second fluid conduit annularly
surrounds a second portion of the first fluid conduit. In some
embodiments, the first fluid contacting chamber conically narrows
in diameter from the top of the fluid contacting chamber to the
exit orfice of the fluid contacting chamber. In some embodiments,
the second fluid conduit and the third fluid conduit are co-axial
for a first portion of the second fluid conduits length. In some
embodiments, the third fluid conduit annularly surrounds a second
portion of the second fluid conduit. In some embodiments, the first
fluid contacting chamber conically narrows in diameter from the top
of the fluid contacting chamber to the exit orfice of the fluid
contacting chamber.
[0009] Some embodiments provide a process for preparing droplets
using an atomizing nozzle apparatus as disclosed and described
herein, comprising applying a first liquid to the first fluid
conduit, applying a second liquid to the second fluid conduit,
applying a third liquid to the third fluid conduit, applying a gas
to the fourth fluid conduit, wherein the gas exiting the fourth
fluid conduit exit orifice impinges the liquid exiting the at least
one exit orifice of the second fluid contacting chamber, providing
atomized droplets, wherein the droplets have an average diameter
from about 100 nm to about 300 .mu.M. In some embodiments, the
first liquid is an emulsion comprised of i) a first aqueous phase,
and ii) a first organic phase comprising a first organic solvent,
the second liquid is a second aqueous phase, and the third liquid
is a second organic phase. In some embodiments, the first organic
solvent is chloroform or methylene chloride. In some embodiments,
the first organic phase further comprises at least one amphipathic
lipid and at least one neutral lipid.
[0010] Some embodiments provide an evaporation apparatus,
comprising at least one atomizing nozzle apparatus of claim 1 and
means for evaporating an organic solvent.
[0011] Some embodiments provide an evaporation apparatus,
comprising a solvent removal vessel having a top, a bottom and a
circular wall, at least one atomizing nozzle connected to the
circular wall, a carrier gas entrance orifice connected to the
circular wall, a solvent removal gas exit orifice centrally
connected to the top, and a product exit orifice connected to the
bottom of the vessel. In some embodiments, at least part of the
solvent removal vessel is jacketed. In some embodiments, the
atomizing nozzle is mounted to and extending through the top of the
solvent removal vessel. In some embodiments, the top of the solvent
removal vessel comprises a lid. In some embodiments, the apparatus
further comprises a rinse nozzle mounted to and extending through
the top of the solvent removal vessel. In some embodiments, the
circular wall has a central axis and the solvent removal gas exit
orifice further comprises a tube extending into the solvent removal
vessel residing along the central axis. In some embodiments, the
atomizing nozzle is angled at least 5 degrees measured off the
central axis of the wall and in a plane parallel to the wall
nearest to it. In some embodiments, the carrier gas entrance
orifice is combined with the atomizing nozzle. In some embodiments,
the solvent removal gas exit orifice further comprises a tube
extending into the solvent removal vessel, wherein the tube is
fitted with a narrowing cone and an annular ring. In some
embodiments, the tube extends from about 1/3 to about 4/5 of the
way into the solvent removal vessel. In some embodiments, the tube
extends about 2/3 of the way into the solvent removal vessel. In
some embodiments, the bottom tip of the narrowing cone of the
solvent removal gas exit orifice diameter is from about 1/1000 to
about 1/5 of a diameter of the inside of the solvent removal
vessel. In some embodiments, the solvent removal gas exit orifice
diameter is less than 1/10 of a diameter of the inside of the
solvent removal vessel. In some embodiments, the at least one
atomizing nozzle is an atomizing nozzle apparatus as disclosed and
described herein. In some embodiments, the ratio of the inside
diameter of the solvent removal vessel to the diameter of the
bottom tip of the narrowing cone of the solvent removal gas exit
orifice is between approximately 5:1 and 100:1. In some
embodiments, the ratio of the inside diameter of the solvent
removal vessel to the diameter of the bottom tip of the narrowing
cone of the solvent removal gas exit orifice is between
approximately 20:1 and 60:1.
[0012] Some embodiments provide a process for preparing large
diameter synthetic membrane vesicles using an evaporation apparatus
as disclosed and described herein, comprising introducing large
diameter synthetic membrane vesicles pre-droplets to the solvent
removal vessel, wherein the large diameter synthetic membrane
vesicles pre-droplets comprise a first component core and an
aqueous phase shell, applying a carrier gas in a tangential
direction to the circular wall through the carrier gas entrance
orifice, and removing a solvent removal gas through the solvent
removal gas exit orifice to provide the large diameter synthetic
membrane vesicles. In some embodiments, the first component core
comprises a first aqueous phase and a first organic phase. In some
embodiments, the first organic phase comprises a continuous first
organic solvent. In some embodiments, the first organic solvent is
chloroform or methylene chloride. In some embodiments, the first
organic phase further comprises at least one amphipathic lipid and
at least one neutral lipid. In some embodiments, the first
component core is first aqueous phase droplets as a suspension in a
first organic phase. In some embodiments, the first aqueous phase
droplets have an average diameter of from about 10 nm to about 10
.mu.m, about 100 nm to about 5 .mu.m, or about 500 nm to about 2
.mu.m. In some embodiments, the first aqueous phase droplets have
an average diameter of about 1 .mu.m. In some embodiments, the
carrier gas comprises nitrogen. In some embodiments, the carrier
gas comprises nitrogen and water vapor. In some embodiments, the
solvent removal gas comprises nitrogen and organic solvent. In some
embodiments, the carrier gas and the solvent removal gas travel in
a vortex in the solvent removal vessel. In some embodiments, the
large diameter synthetic membrane vesicles are multivesicular
liposomes having a structure including multiple non-concentric
chambers and comprising at least one amphipathic lipid and at least
one neutral lipid. In some embodiments, the at least one
amphipathic lipid is selected from the group consisting of
phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine. In some embodiments, the at least one
amphipathic lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol, and
1,2-dioleoyl-sn-glycero-3-phosphoglycerol. In some embodiments, the
at least one neutral lipid is selected from the group consisting of
glycerol esters, glycol esters, tocopherol esters, sterol esters,
hydrocarbons and squalenes. In some embodiments, the at least one
neutral lipid is selected from the group consisting of triolein,
tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin. In some embodiments, the
multivesicular liposomes further comprises a therapeutic agent. In
some embodiments, the therapeutic agent is bupivacaine. In some
embodiments, the multivesicular liposomes comprises an outer
surface layer whose composition is different than the composition
of the internal structure.
[0013] Some embodiments provide an evaporation apparatus,
comprising at least one atomizing nozzle apparatus as disclosed and
described herein and means for removing an organic solvent from a
droplet.
[0014] Some embodiments provide a process for preparing large
diameter synthetic membrane vesicles using the evaporation
apparatus as disclosed and described herein, comprising introducing
large diameter synthetic membrane vesicles pre-droplets to the
solvent removal vessel, wherein the large diameter synthetic
membrane vesicles pre-droplets comprise a first component core and
an aqueous phase shell; applying a carrier gas in a tangental
direction to the circular wall through the carrier gas entrance
orifice; removing a solvent removal gas through the solvent removal
gas exit orifice to provide pre-temperature treatment large
diameter synthetic membrane vesicles; introducing the
pre-temperature treatment large diameter synthetic membrane
vesicles to an outlet line; contacting the pre-temperature
treatment large diameter synthetic membrane vesicles with a hot
solution in the outlet line, wherein the hot solution has a
temperature ranging from about 30.degree. C. to about 100.degree.
C. to provide post-temperature treatment large diameter synthetic
membrane vesicles; transferring the post-temperature treatment
large diameter synthetic membrane vesicles to a continuous-flow
particle-concentration system or continuous phase exchange system;
cooling the post-temperature treatment large diameter synthetic
membrane vesicles to a second temperature to provide the large
diameter synthetic membrane vesicles; and isolating the large
diameter synthetic membrane vesicles. In some embodiments, the
first component core comprises a first aqueous phase and a first
organic phase. In some embodiments, the first organic phase
comprises a continuous first organic solvent. In some embodiments,
the first organic solvent is chloroform or methylene chloride. In
some embodiments, the first organic phase further comprises at
least one amphipathic lipid and at least one neutral lipid. In some
embodiments, the first component core is a suspension of first
aqueous phase droplets in a first organic phase. In some
embodiments, the first aqueous phase droplets have an average
diameter of from about 10 nm to about 10 .mu.m, about 100 nm to
about 5 .mu.m, or about 500 nm to about 2 .mu.m. In some
embodiments, the first aqueous phase droplets have an average
diameter of about 1 .mu.m. In some embodiments, the carrier gas
comprises nitrogen. In some embodiments, the solvent removal gas
comprises nitrogen and organic solvent. In some embodiments, the
carrier gas and the solvent removal gas travel in a vortex in the
solvent removal vessel. In some embodiments, the large diameter
synthetic membrane vesicles are multivesicular liposomes having a
structure including multiple non-concentric chambers and comprising
at least one amphipathic lipid and at least one neutral lipid. In
some embodiments, the at least one amphipathic lipid is selected
from the group consisting of phosphatidylcholines,
phosphatidylserines, phosphatidylethanolamines,
phosphatidylinositols, sphingomyelin, soybean lecithin (soya
lecithin), egg lecithin, lysophosphatidylcholines,
lysophosphatidylethanolamines, phosphatidylglycerols,
phosphatidylserines, phosphatidylinositols, phosphatidic acids,
cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine. In some embodiments, the at least one
amphipathic lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol, and
1,2-dioleoyl-sn-glycero-3-phosphoglycerol. In some embodiments, the
at least one neutral lipid is selected from the group consisting of
glycerol esters, glycol esters, tocopherol esters, sterol esters,
hydrocarbons and squalenes. In some embodiments, the at least one
neutral lipid is selected from the group consisting of triolein,
tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin. In some embodiments, the
multivesicular liposomes further comprises a therapeutic agent. In
some embodiments, the therapeutic agent is bupivacaine.
[0015] Some embodiments provide a composition comprising
multivesicular liposomes having a structure including multiple
non-concentric chambers and comprising at least one amphipathic
lipid and at least one neutral lipid, wherein said multivesicular
liposomes are made by a process comprising removing organic solvent
from multivesicular liposomes pre-droplets using an evaporation
apparatus as disclosed and described herein, said process
comprising introducing multivesicular liposomes pre-droplets
pre-droplets to the solvent removal vessel, wherein the large
diameter synthetic membrane vesicles pre-droplets comprise a first
component core and an aqueous phase shell; applying a carrier gas
in a tangental direction to the circular wall through the carrier
gas entrance orifice; and removing a solvent removal gas through
the solvent removal gas exit orifice to provide the large diameter
synthetic membrane vesicles.
[0016] Some embodiments provide a composition comprising large
diameter synthetic membrane vesicles made by a process disclosed
and described herein. In some embodiments, the large diameter
synthetic membrane vesicles are multivesicular liposomes having a
structure including multiple non-concentric chambers and comprising
at least one amphipathic lipid and at least one neutral lipid. In
some embodiments, the at least one amphipathic lipid is selected
from the group consisting of phosphatidylcholines,
phosphatidylserines, phosphatidylethanolamines,
phosphatidylinositols, sphingomyelin, soybean lecithin (soya
lecithin), egg lecithin, lysophosphatidylcholines,
lysophosphatidylethanolamines, phosphatidylglycerols,
phosphatidylserines, phosphatidylinositols, phosphatidic acids,
cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, and ethyl
phosphatidylcholine. In some embodiments, the at least one
amphipathic lipid is selected from the group consisting of
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol, and
1,2-dioleoyl-sn-glycero-3-phosphoglycerol. In some embodiments, the
at least one neutral lipid is selected from the group consisting of
glycerol esters, glycol esters, tocopherol esters, sterol esters,
hydrocarbons and squalenes. In some embodiments, the at least one
neutral lipid is selected from the group consisting of triolein,
tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin. In some embodiments, the
multivesicular liposomes further comprises a therapeutic agent. In
some embodiments, the therapeutic agent is bupivacaine.
[0017] Some embodiments provide a continuous-flow emulsification
system, comprising a mixer, comprised of a rotor and a stator; a
recirculation loop, comprised of one or more recirculation lines; a
heat exchanger; one or more outlet lines; one or more continuous
phase inlet lines; and a discontinuous phase inlet line, wherein
the heat exchanger and the mixer are connected together in the
recirculation loop by one or more recirculation lines; further
wherein the one or more outlet lines and one or more continuous
phase inlet lines are connected to the recirculation loop; further
wherein the end of the discontinuous phase inlet line is located
within approximately 1/3rd of a rotor diameter from the rotor and
approximately 1/3rd of a rotor diameter of the rotation axis of the
rotor and is in fluid communication with the rotor. In some
embodiments, a continuous phase entrance line is connected to the
recirculation loop upstream of the mixer and downstream of the heat
exchanger and an outlet line is connected to the recirculation loop
downstream of the mixer and upstream of the heat exchanger. In some
embodiments, the continuous-flow emulsification system further
comprises an emulsion, which emulsion recirculates through the
recirculation line back to the mixer an average of at least 5 or
more times. In some embodiments, the continuous-flow emulsification
system further comprises emulsion droplets produced in the mixer,
which are on average less than 10 microns in diameter.
[0018] Some embodiments provide a continuous processing system,
comprising one or more concentrator units, each unit comprising a
retentate vessel; a particle suspension inlet line, connected to
the retentate vessel; a first outlet line, connecting the retentate
vessel and a particle concentrating device; a pump located along
the first outlet line between the retentate vessel and the particle
concentrating device; and a second outlet line, leading to another
concentrator unit or the final product collection vessel; and means
for removing or exchanging solvent. In some embodiments, the means
for removing or exchanging solvent are each independently selected
from the group consisting of a tangential flow filtration unit, a
hydro-cyclone unit, and a centrifugal separator. In some
embodiments, the continuous processing system further comprises a
new suspending medium inlet line connected to the retentate vessel.
In some embodiments, the means for removing or exchanging solvent
are each a tangential flow filtration unit. In some embodiments,
the means for removing or exchanging solvent are each a centrifugal
separator. In some embodiments, the system comprises at least one
tangential flow filtration unit and at least one centrifugal
separator. In some embodiments, the continuous processing system is
a continuous-flow particle-concentration system or continuous phase
exchange system.
[0019] Some embodiments provide a process for making multivesicular
liposomes using the atomizing nozzle apparatus as disclosed and
described herein, comprising applying a first liquid to the first
fluid conduit, wherein the first liquid comprises an organic
solvent, applying a second liquid to the second fluid conduit,
applying a pressurized gas to the third fluid conduit to provide
atomized droplets, wherein the pressurized gas exiting the third
fluid conduit exit orifice impinges the liquid exiting the fluid
contacting chamber exit orifice, and removing the organic solvent
from the atomized droplets, wherein less than 4000 ppm of the
organic solvent remains in the atomized droplets. In some
embodiments, the first liquid is an emulsion comprised of a
discontinuous aqueous phase, and a continuous organic phase
comprising the organic solvent. In some embodiments, the organic
solvent is methylene chloride. In some embodiments, the continuous
organic phase further comprises a therapeutic agent. In some
embodiments, the therapeutic agent is bupivacaine or a salt
thereof. In some embodiments, the second liquid applied to the
second fluid conduit is an aqueous solution. In some embodiments,
the aqueous solution further comprises dextrose and lysine. In some
embodiments, the gas is a sterilized gas. In some embodiments, the
gas is nitrogen. In some embodiments, the process further comprises
introducing atomized droplets to an evaporation apparatus as
disclosed and described herein; introducing a pressurized carrier
gas tangentially to the circular wall into the solvent removal
vessel through the carrier gas entrance orifice; removing a solvent
removal gas wherein the solvent removal gas removes greater than
90% of the organic solvent in the atomized droplets resulting in
formation of multivesicular liposomes. In some embodiments, the
carrier gas is heated and humidified. In some embodiments, the
process further comprises spraying a wall rinse solution into the
solvent removal vessel using a rinse nozzle, wherein the wall rinse
solution prevents build-up of particles in the evaporation
apparatus. In some embodiments, the atomized droplets contain
organic solvent in the range of from about 400 ppm to about 3500
ppm.
[0020] Some embodiments provide a process for making an emulsion
using an emulsification system as disclosed and described herein,
comprising feeding an organic discontinuous phase into the
emulsification system through the discontinuous phase inlet line
and feeding an aqueous continuous phase into the emulsification
system through one or more continuous phase inlet lines.
[0021] Some embodiments provide a process for making an emulsion
using the emulsification system as disclosed and described herein,
comprising feeding an aqueous discontinuous phase into the
emulsification system through the discontinuous phase inlet line
and feeding an organic continuous phase fed into the emulsification
system through the one or more continuous phase inlet lines. In
some embodiments, the organic continuous phase is comprised of an
organic solvent and a neutral lipid. In some embodiments, the
organic solvent is methylene chloride. In some embodiments, the
aqueous discontinuous phase is comprised of an acid and a
therapeutic agent. In some embodiments, the acid is phosphoric
acid. In some embodiments, the therapeutic agent is bupivacaine. In
some embodiments, a portion of the emulsion is fed through one or
more outlet lines to the inner fluid conduit of an atomizing nozzle
as disclosed and described herein. In some embodiments, a portion
of the emulsion is fed through one or more outlet lines to an
evaporation apparatus as disclosed and described herein.
[0022] Some embodiments provide a process for preparing large
diameter synthetic membrane vesicles using an evaporation apparatus
as disclosed and described herein.
[0023] Some embodiments provide a large diameter synthetic membrane
vesicles made by a process as disclosed and described herein.
[0024] Some embodiments produce a plurality of MVL particles made
by using the process utilizing the atomizing nozzle disclosed
herein.
[0025] Some embodiments produce a plurality of MVL particles made
by using the process utilizing the emulsification system and the
atomizing nozzle disclosed herein.
[0026] Some embodiments produce a plurality of MVL particles made
by using the process utilizing the evaporation apparatus and the
atomizing nozzle disclosed herein.
[0027] Some embodiments produce a plurality of MVL particles made
by using the process utilizing the emulsification system, the
evaporation apparatus and the atomizing nozzle disclosed
herein.
[0028] Some embodiments produce a plurality of MVL particles made
by using the process utilizing the evaporation apparatus, the
atomizing nozzle, and particle concentration system disclosed
herein.
[0029] Some embodiments produce a plurality of MVL particles made
by using the process utilizing the emulsification system, the
evaporation apparatus, the atomizing nozzle, and particle
concentration system disclosed herein.
[0030] Some embodiments produce a plurality of MVL particles made
by any of the above product-by-process embodiments, wherein the MVL
contains bupivacaine or a pharmaceutically acceptable salt
thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] These and other objects and features of the instant
embodiment will become more fully apparent in the following
descriptions and appended claims taken in conjuction with the
following drawings where like references numbers indicate identical
or functionally similar elements.
[0032] FIG. 1A is a schematic diagram of significant components
used in one of the instant systems for manufacturing synthetic
membrane vesicles.
[0033] FIG. 1B is a schematic diagram of significant components
used in another system for manufacturing synthetic membrane
vesicles.
[0034] FIG. 1C is a schematic of one embodiment of a continuous
heat treatment system including a temperature controlled tank and a
holding coil tubing used in a manufacturing system such as the
manufacturing system of FIG. 1A or FIG. 1B.
[0035] FIG. 2 is a schematic of one embodiment of an emulsification
system used in the manufacturing system of FIG. 1A or FIG. 1B.
[0036] FIG. 3A is a schematic view of one embodiment of a three
channel atomizing nozzle used in a manufacturing system, such as
the manufacturing system of FIG. 1A of FIG. 1B.
[0037] FIG. 3B is a cross-sectional view of the atomizing nozzle of
FIG. 3A, taken along line B of FIG. 3A.
[0038] FIG. 3C is an expanded view of the cylindrical tip of the
atomizing nozzle of FIG. 3A, shown as position C in FIG. 3A.
[0039] FIG. 3D is an expanded view of an atomized droplet produced
by the atomizing nozzle of FIG. 3A, shown as position D in FIG.
3A.
[0040] FIG. 3E is a schematic view of one embodiment of a four
channel atomizing nozzle used in a manufacturing system, such as
the manufacturing system of FIG. 1A or FIG. 1B.
[0041] FIG. 3F is a cross-sectional view of the atomizing nozzle of
FIG. 3E, taken along line F of FIG. 3E.
[0042] FIG. 3G is an expanded view of the cylindrical tip of the
atomizing nozzle of FIG. 3E, shown as position G in FIG. 3E.
[0043] FIG. 3H is an expanded view of an atomized droplet produced
by the atomizing nozzle of FIG. 3E, shown as position H in FIG.
3E.
[0044] FIG. 3I is a schematic view of another embodiment of a four
channel atomizing nozzle used in a manufacturing system, such as
the manufacturing system of FIG. 1A or FIG. 1B.
[0045] FIG. 3J is a cross-sectional view of the atomizing nozzle of
FIG. 3I, taken along line J of FIG. 3I.
[0046] FIG. 3K is an expanded view of the cylindrical tip of the
atomizing nozzle of FIG. 3I, shown as position K in FIG. 3I.
[0047] FIG. 3L is an expanded view of an atomized droplet produced
by the atomizing nozzle of FIG. 3I, shown as position L in FIG.
3I.
[0048] FIG. 4A is a schematic view of one embodiment of an
atomizing nozzle used in a manufacturing system, such as the
manufacturing system of FIG. 1A or FIG. 1B.
[0049] FIG. 4B is a cross-sectional view of the atomizing nozzle of
FIG. 4A, taken along line B of FIG. 4A.
[0050] FIG. 4C is an expanded view of the cylindrical tip of the
atomizing nozzle of FIG. 4A, shown as position C in FIG. 4A.
[0051] FIG. 4D is an expanded view of an atomized droplet produced
by the atomizing nozzle of FIG. 4A, shown as position D in FIG.
4A.
[0052] FIG. 5 is a detailed view of one embodiment of an atomizing
nozzle used in a manufacturing system, such as the manufacturing
system of FIG. 1A or FIG. 1B.
[0053] FIG. 6 is an exploded view of the individual components of
the atomizing nozzle of FIG. 5.
[0054] FIG. 7 is a schematic of one embodiment of a solvent removal
vessel used for evaporating solvent from atomized particles in a
manufacturing system such as the manufacturing system of FIG. 1A or
FIG. 1B.
[0055] FIG. 8 is a schematic of one embodiment of a particle
concentration system used in a manufacturing system such as the
manufacturing system of FIG. 1A or FIG. 1B.
[0056] FIG. 9A and FIG. 9B provide cross-sectional views of a
droplet, a more detailed view of a droplet, and a large diameter
synthetic membrane vesicle particle. In some embodiments, the large
diameter synthetic membrane vesicle particle can be a MVL particle.
The large diameter synthetic membrane vesicle particle can be
formed by removal of the organic solvent from the emulsion
droplet.
[0057] FIG. 10 is a schematic of one embodiment of a particle
concentration system including a plurality of filtration units and
at least one centrifuge unit used in a manufacturing system such as
the manufacturing system of FIG. 1A or FIG. 1B.
[0058] FIG. 11 is a schematic of one embodiment of a particle
concentration system including a plurality of centrifuge units used
in a manufacturing system such as the manufacturing system of FIG.
1A or FIG. 1B.
[0059] FIG. 12 is a graph of an in vitro release study of
multivesicular liposome formulations.
[0060] FIG. 13 is a graph of an in vivo PK study of multivesicular
liposome formulations.
[0061] FIG. 14 is a graph of an accelerated stability profile study
of a multivesicular formulation made with the instant manufacturing
system with and without heat treatment. The low osmolality is more
stable than the sample without heat treatment and the heat treated
sample is most stable.
[0062] FIG. 15 is a graph of an in-vivo release profile study of a
multivesicular formulation made with and without heat
treatment.
[0063] Large scale methods of manufacturing large diameter
synthetic membrane vesicles, such as multivesicular liposomes,
often require large amounts of solvents, time sensitive steps and
concentration adjustment of the final product under sterile
conditions, for example, as described in WO99/25319. In addition,
current methods of manufacturing large diameter synthetic membrane
vesicles, such as multivesicular liposomes on a commercial scale,
require significant commitments in manufacturing space, cost, and
time. As such, developing stable multivesicular liposome
formulations containing a therapeutic agent in a cost effective and
timely manner remains an ongoing challenge.
[0064] In large scale manufacturing of large diameter synthetic
membrane vesicles, the present embodiments require less water for
injection (WFI), space, time and energy to produce an equivalent
amount of the large diameter synthetic membrane vesicles than under
previously described large scale manufacturing conditions. As a
result, waste disposal is reduced and overall costs are reduced.
For example, the present embodiments provide systems that can be
housed in a one story room (e.g. 5.times.5 meter room) that
previously described systems would require a multistory room with
at least a ten fold increase in the area of the room. Additionally,
the present embodiments provide systems that are particularly well
adapted to continuous processing for more rapid production of large
diameter synthetic membrane vesicles and allowing for more
efficient implementation of clean-in-place (CIP) and
sterile-in-place (SIP) protocols. Allowing for greatly reduced
utility requirements to implement CIP and SIP protocols.
[0065] The devices described in FIGS. 1A and 1B are particularly
well suited for making multivesicular liposomes (MVL).
Multivesicular liposomes (MVL), first reported by Kim, et al.
(Biochim, Biophys. Acta, 728:339-348, 1983), are uniquely different
from other lipid-based drug delivery systems such as unilamellar
(Huang, Biochemistry, 8:334-352, 1969; Kim, et al., Biochim.
Biophys. Acta, 646:1-10, 1981) and multilamellar (Bangham, et al.,
J. Mol. Bio., 13:238-252, 1965) liposomes. For example,
multivesicular liposomes made by the processes described herein
typically can have diameters ranging from about 10 to 100 .mu.m,
and more typically ranging from about 20 to 55 .mu.m. In contrast,
multilamellar liposomes usually have diameters of from 0.2 to 5
.mu.m and unilamellar liposomes usually have diameters of from 0.02
to 0.5 .mu.m.
[0066] Additionally, multivesicular liposomes (MVL) contain
multiple aqueous chambers per particle and the multiple aqueous
chambers are non-concentric. In contrast unilamellar liposomes
(also known as unilamellar vesicles) and multilamellar liposomes
(also known as multilamellar vesicles) contain a single chamber per
particle. Further, neutral lipids are necessary to form
multivesicular liposomes (MVL). In contrast unilamellar liposomes
(also known as unilamellar vesicles) and multilamellar liposomes
(also known as multilamellar vesicles) do not require inclusion of
neutral lipids to form.
[0067] Multivesicular liposomes (MVL) are entirely distinct from
unilamellar liposomes and multilamellar liposomes. The structural
and functional characteristics of multivesicular liposomes are not
directly predictable from current knowledge of unilamellar
liposomes and multilamellar liposomes. As described in the book
edited by Jean R. Philippot and Francis Schuber (Liposomes as Tools
in Basic Research and Industry, CRC press, Boca Raton, Fla., 1995,
page 19), Multivesicular liposomes (MVL) are bounded by an external
bilayer membrane shell, but have a very distinctive internal
morphology, which may arise as a result of the special method
employed in the manufacture. Topologically, multivesicular
liposomes (MVL) are defined as liposomes containing multiple
non-concentric chambers within each liposome particle, resembling a
"foam-like" matrix; whereas multilamellar vesicles contain multiple
concentric chambers within each liposome particle, resembling the
"layers of an onion".
[0068] The presence of internal membranes distributed as a network
throughout multivesicular liposomes (MVL) may serve to confer
increased mechanical strength to the vesicle, while still
maintaining a high volume:lipid ratio compared with multilamellar
vesicles. The multivesicular nature of multivesicular liposomes
(MVL) also indicates that, unlike for unilamellar liposomes, a
single breach in the external membrane of a multivesicular
liposomes (MVL) will not result in total release of the internal
aqueous contents. Thus, both structurally and functionally the
multivesicular liposomes (MVL) are unusual, novel and distinct from
all other types of liposomes. As a result, the functional
properties of multivesicular liposomes (MVL) are not predictable
based on the prior art related to conventional liposomes such as
unilamellar liposomes and multilamellar liposomes.
[0069] In some embodiments, the large diameter synthetic membrane
vesicles are multivesicular liposomes. In some embodiments, the
multivesicular liposomes further comprise bupivaciane, DEPC, DPPG,
and tricaprylin. In some embodiments, the multivesicular liposomes
further comprise bupivacaine phosphate, DEPC, DPPG, and
tricaprylin. In some embodiments, the multivesicular liposomes
further comprise bupivacaine, DEPC, DPPG, tricaprylin and
cholesterol. In some embodiments, the multivesicular liposomes
further comprise bupivacaine phosphate, DEPC, DPPG, tricaprylin and
cholesterol.
[0070] In some embodiments, the multivesicular liposomes further
comprise bupivaciane, dextrose, L-Lysine, DEPC, DPPG, and
tricaprylin. In some embodiments, the multivesicular liposomes
further comprise bupivacaine phosphate, dextrose, L-Lysine, DEPC,
DPPG, and tricaprylin. In some embodiments, the multivesicular
liposomes further comprise bupivacaine, dextrose, L-Lysine, DEPC,
DPPG, tricaprylin and cholesterol. In some embodiments, the
multivesicular liposomes further comprise bupivacaine phosphate,
dextrose, L-Lysine, DEPC, DPPG, tricaprylin and cholesterol.
[0071] In some embodiments, the multivesicular liposomes further
comprise bupivaciane, dextrose, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, dextrose, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine, dextrose, DEPC, DPPG, tricaprylin and cholesterol. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, dextrose, DEPC, DPPG, tricaprylin and
cholesterol.
[0072] In some embodiments, the multivesicular liposomes further
comprise bupivaciane, L-Lysine, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, L-Lysine, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine, L-Lysine, DEPC, DPPG, tricaprylin and cholesterol. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, L-Lysine, DEPC, DPPG, tricaprylin and
cholesterol.
[0073] In some embodiments, the multivesicular liposomes further
comprise bupivacaine, morphine, cytarabine, or their
pharmaceutically acceptable salts as the therapeutic agent. In some
embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, morphine sulfate, or cytarabine HCl.
[0074] In another embodiment, any one of the above described
embodiments can be used alone or in combination with any one or
more of the above described embodiments. For example, any above
described atomizing nozzle, evaporation apparatus, continuous-flow
emulsification system, continuous-flow diafiltration system,
continuous-flow diafiltration further comprising one or more
centrifuges, continuous-flow centrifuge system, or continuous
processing system can be used alone or in combination. Thus, an
evaporation apparatus can be used in conjunction with a three-fluid
atomizing nozzle. This evaporation system/atomizing nozzle can be
used with a continuous-flow emulsification system, as depicted in
FIGS. 1A, 1B, and 1C. The three-fluid atomizing nozzle/evaporation
apparatus combination can be used in conjunction with a
continuous-flow system, as depicted in FIGS. 8, 10, and 11. Any of
these combinations can be used to make multivesicular liposomes. In
particular any of the combinations can be used to make
multivesicular liposomes containing bupivacaine or its salts as the
therapeutic agent.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
[0075] Features of the present disclosure will become more fully
apparent from the following description and appended claims, taken
in conjunction with the accompanying drawings. It will be
understood these drawings depict only certain embodiments in
accordance with the disclosure and, therefore, are not to be
considered limiting of its scope; the disclosure will be described
with additional specificity and detail through use of the
accompanying drawings. An apparatus, system or method according to
some of the described embodiments can have several aspects, no
single one of which necessarily is solely responsible for the
desirable attributes of the apparatus, system or method. After
considering this discussion, and particularly after reading the
section entitled "Detailed Description of the Preferred Embodiment"
one will understand how illustrated features serve to explain
certain principles of the present disclosure.
[0076] In the following detailed description, only certain
exemplary embodiments have been shown and described, simply by way
of illustration. As those skilled in the art would realize, the
described embodiments may be modified in various different ways,
all without departing from the spirit or scope of the present
disclosure. Accordingly, the drawings and description are to be
regarded as illustrative in nature and not restrictive. In
addition, when an element is referred to as being "on" another
element, it can be directly on the another element or be indirectly
on the another element with one or more intervening elements
interposed therebetween. Also, when an element is referred to as
being "connected to" another element, it can be directly connected
to the another element or be indirectly connected to the another
element with one or more intervening elements interposed
therebetween. Hereinafter, like reference numerals refer to like
elements. Since the disclosure may be modified in various ways and
have various embodiments, the disclosure will be described in
detail with reference to the drawings. However, it should be
understood that the disclosure is not limited to a specific
embodiment but includes all changes and equivalent arrangements and
substitutions included in the spirit and scope of the disclosure.
In the following description, if the detailed description of the
already known structure and operation may confuse the subject
matter of the present disclosure, the detailed description thereof
will be omitted.
[0077] While such terms as "first," "second," etc., may be used to
describe various components, such components must not be limited to
the above terms. The above terms are used only to distinguish one
component from another. Terms used in the following description are
to describe specific embodiments and is not intended to limit the
disclosure. The expression of singularity includes plurality
meaning unless the singularity expression is explicitly different
in context. It should be understood that the terms "comprising,"
"having," "including," and "containing" are to indicate features,
numbers, steps, operations, elements, parts, and/or combinations
but not to exclude one or more features, numbers, steps,
operations, elements, parts, and/or combinations or additional
possibilities.
[0078] Some embodiments provide continuous processes for making
multivesicular liposomes. Prior methods of making multivesicular
liposomes required batch processing. This batch processing required
removal of the solvent from the droplets of first emulsion
surrounded by a second aqueous phase by contacting the suspension
of first emulsion droplets in a continuous aqueous phase with a
discontinuous gas phase by sparging (bubbling) gas through the
aqueous phase or blowing gas over a flask containing continuous
aqueous phase. This batch processing takes tens of minutes to
remove the solvent.
[0079] It was surprisingly discovered that forming a first emulsion
surrounded by an aqueous shell in the form of a droplet and
contacting it with a continuous gas phase, reduces the time needed
to remove the organic solvent to a few seconds and possibly a
fraction of a second. (much less than the tens of minutes stated
above for batch processing).
[0080] This is due to the tremendous gas contacting surface area of
the atomized droplets; and the much faster diffusion of the solvent
in gasses versus water; and the very short distances that the
solvent has to diffuse through the aqueous phase to reach the gas
(now only microns instead of the distanced between sparging
bubbles.)
[0081] This extremely fast solvent removal enables the continuous
processing. The solvent is removed in less than the time that it
takes for the atomized droplets to reach the bottom of the solvent
removal vessel (a few seconds at most).
DEFINITIONS
[0082] As used herein, abbreviations are defined as follows: [0083]
aq. Aqueous [0084] CIP Clean-in-place processing [0085] .degree. C.
Temperature in degrees Celsius [0086] d10 the diameter where 10
mass-% (volume %) (of the particles) of the particles have a
smaller equivalent diameter, and the other 90 mass-% (volume %)
have a larger equivalent diameter in .mu.m [0087] d50 the diameter
where 50 mass-% (volume %) (of the particles) of the particles have
a smaller equivalent diameter, and the other 50 mass-% (volume %)
have a larger equivalent diameter in .mu.m [0088] d90 the diameter
where 90 mass-% (volume %) (of the particles) of the particles have
a smaller equivalent diameter, and the other 100 mass-% (volume %)
have a larger equivalent diameter in .mu.m [0089] DCM methylene
chloride [0090] g Gram(s) [0091] h Hour (hours) [0092] mL
Milliliter(s) [0093] mg Milligram(s) [0094] mOsm/kg Osmolality per
kilogram [0095] pH measure of the acidity or alkalinity of a liquid
using a pH meter or pH indicator [0096] PPV Packed particle volume
[0097] PSD Particle size distribution [0098] RT, rt Room
temperature [0099] SIP Sterile-in-place processing [0100] Tert, t
tertiary [0101] .mu.L Microliter(s) [0102] .mu.g Microgram(s)
[0103] WFI Water for injection
[0104] Some embodiments relate to a process for preparing a large
diameter synthetic membrane vesicle composition comprising
preparing a first component by mixing a first aqueous phase and an
organic phase, said organic phase comprising an organic solvent, at
least one amphipathic lipid and at least one neutral lipid;
preparing a droplet(s) by mixing said first component and a second
aqueous phase, said droplet(s) comprising an aqueous phase;
preparing a large diameter synthetic membrane vesicle by removing
the organic solvent from the droplet(s), wherein the removing
comprises contacting the droplet(s) with a gas; and collecting the
large diameter synthetic membrane vesicle particles, wherein the
large diameter synthetic membrane vesicle is suspended in a
continuous aqueous phase.
[0105] Some embodiments relate to a process for preparing a
multivesicular liposome composition comprising preparing a first
component by mixing a first aqueous phase and an organic phase,
said organic phase comprising an organic solvent, at least one
amphipathic lipid and at least one neutral lipid, wherein the first
component comprises a therapeutic agent; preparing a droplet(s) by
mixing said first component and a second aqueous phase, said
emulsion droplet(s) comprising an aqueous phase; preparing a
multivesicular liposome particle by removing the organic solvent
from the emulsion droplet, wherein the removing comprises
contacting the emulsion droplet with a gas; and preparing a
multivesicular liposome composition by collecting the
multivesicular liposome particles, wherein the multivesicular
liposome composition is suspended in a continuous aqueous
phase.
[0106] Some embodiments relate to a process for preparing a
multivesicular liposome composition comprising preparing a first
component by mixing a first aqueous phase and an organic phase,
said organic phase comprising an organic solvent, at least one
amphipathic lipid and at least one neutral lipid, wherein the first
component comprises a therapeutic agent, preparing an emulsion of
first component droplets, a second component droplet, by mixing
said first component and a second aqueous phase, said second
component droplet comprising an aqueous phase, wherein the second
component droplet is prepared using a device as described herein,
preparing a multivesicular liposome particle by removing the
organic solvent from the second component droplet, wherein the
removing comprises contacting the second component droplet with a
gas, and preparing a multivesicular liposome composition by
collecting the multivesicular liposome particles, wherein the
multivesicular liposome composition is suspended in a continuous
aqueous phase.
[0107] As used herein the term "amphipathic lipid" refers to a
substance including a hydrophilic region and a hydrophobic region,
such as phospholipids. Amphipathic lipids can be zwitterionic
phospholipids, zwitterionic lipids, lipids having a net negative
charge, and lipids having a net positive charge. For example,
amphipathetic lipids include, but are not limited to,
phosphatidylcholines, phosphatidylserines,
phosphatidylethanolamines, phosphatidylinositols, sphingomyelin,
soybean lecithin (soya lecithin), egg lecithin,
lysophosphatidylcholines, lysophosphatidylethanolamines,
phosphatidylglycerols, phosphatidylserines, phosphatidylinositols,
phosphatidic acids, cardiolipins, acyl trimethylammonium propane,
diacyldimethylammonium propane, stearylamine, ethyl
phosphatidylcholine and the like. Phospholipids used in the methods
described herein can be of a single class or a mixture of classes.
Some embodiments include crude preparations of phospholipids, such
as soybean lecithin (soya lecithin) and egg lecithin. Soya lecithin
is a combination predominantly of naturally-occurring
phospholipids; phosphatidylcholine (PC), phosphatidylethanolamine
(PE) and phosphatidylinositol (PI). Examples of
phosphatidylcholines and phosphatidylglycerols include, but are not
limited to, 1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol, and mixtures
thereof.
[0108] As used herein the term "neutral lipid" refers to oils,
waxes or fatty acid esters that lack a charged or hydrophilic head
group. Neutral lipids include but are not limited to, glycerol
esters, glycol esters, tocopherol esters, sterol esters,
hydrocarbons and squalenes.
[0109] As used herein the terms "glycerol esters," "triglycerides,"
and "triacylglycerols" refer to triesters formed from glycerol and
fatty acids. Glycerol esters include but are not limited to,
triolein, tripalmitolein, trimyristolein, trilinolein, tributyrin,
tricaprylin, tricaproin, and tricaprin.
[0110] As used herein the term "organic solvent" refers to ethers,
esters, halogenated ethers, aromatic or aliphatic hydrocarbons,
aromatic or aliphatic halohydrocarbons, or Freons. Organic solvents
include but are not limited to, diethyl ether, tert-butylmethyl
ether, tetrahydrofuran, sevoflurane, desflurane, isoflurane,
enflurane, halothane, chloroform, dichloromethane, ethyl acetate,
hexane, hexanes, cyclohexane, pentane, cyclopentane, petroleum
ether, toluene, and any combinations thereof.
[0111] As used herein the term "aqueous phase" refers to any
solution or mixture having water as the major component. The
aqueous phase can include constituents, such as pH buffering
agents, salts, osmotic agents, simple sugars, amino acids,
electrolytes, preservatives, other water soluble excipients and the
like. Aqueous phase constituents can include, but are not limited
to, sodium chloride, hydrochloric acid, phosphoric acid, lysine,
dextrose, glucose and the like.
[0112] As used herein the term "fluid" refers to a substance that
has the ability to flow. Fluids can be a gas, a liquid, a liquid
with a substance(s) suspended throughout the liquid, an emulsion, a
vapor, or a gas/vapor mixture.
[0113] As used herein the term "therapeutic agent" and "drug"
refers to a chemical compound, mixtures of chemical compounds, or
biological molecules, such as biological macromolecules or peptides
that may have therapeutic properties. The therapeutic agent can be
purified, substantially purified or partially purified. In some
embodiments, the therapeutic agent can be selected from the group
including antianginas, antiarrhythmics, antiasthmatic agents,
antibiotics, antidiabetics, antifungals, antihistamines,
antihypertensives, antiparasitics, antineoplastics, antitumor
drugs, antivirals, cardiac glycosides, hormones, immunomodulators,
monoclonal antibodies, neurotransmitters, nucleic acids, proteins,
radio contrast agents, radionuclides, sedatives, analgesics,
steroids, tranquilizers, vaccines, vasopressors, anesthetics,
peptides and the like. Any pharmaceutically acceptable salt of a
particular therapeutic agent is also envisioned as being useful in
the present embodiments. In some embodiments, the therapeutic agent
can be introduced in either an aqueous or a solvent phase,
depending on their solubility in these phases. In a typical
embodiment, the therapeutic agent can be selected from the group
including semisynthetic aminoglycoside antibiotics such as
amikacin; antidiabetics; peptides such as insulin; antitumor drugs
such as paclitaxel; antineoplastics including cytarabine,
5-fluorouracil and floxuridine; alkaloid opiate analgesics
including morphine and hydromorphine; local anesthetics including
bupivacaine; synthetic anti-inflamniatory adrenocortical steroids
including dexamethasone; antimetabolites including methotrexate;
glycopeptide antibiotics including bleomycin; vincaleukoblastines
and stathmokinetic oncolytic agents including vincristine and
vinblastine; hormones, plasma proteins, cytokines, growth factors,
DNA and RNA from a variety of organisms, and antisense
oligonucleotides. In some embodiments, the therapeutic agent can be
an amide anesthetic. Amide anesthetics include, but are not limited
to, bupivacaine, mepivacaine, ropivacaine, lidocaine, pyrrocaine,
prilocalne, their stereoisomers, and combinations thereof, and
pharmaceutically acceptable salts thereof.
[0114] As used herein the term "pharmaceutically acceptable salt"
refers to a salt of a compound that does not cause significant
irritation to an organism to which it is administered and does not
abrogate the biological activity and properties of the compound. In
some embodiments, the salt is an acid addition salt of the
compound. Pharmaceutical salts can be obtained by reacting a
compound with inorganic acids such as hydrohalic acid (e.g.,
hydrochloric acid or hydrobromic acid), sulfuric acid, nitric acid,
phosphoric acid and the like. Pharmaceutical salts can also be
obtained by reacting a compound with an organic acid such as
aliphatic or aromatic carboxylic or sulfonic acids, for example
acetic, succinic, lactic, malic, tartaric, citric, ascorbic,
nicotinic, methanesulfonic, ethanesulfonic, p-toluensulfonic,
salicylic or naphthalenesulfonic acid. Pharmaceutical salts can
also be obtained by reacting a compound with a base to form a salt
such as an ammonium salt, an alkali metal salt, such as a sodium or
a potassium salt, an alkaline earth metal salt, such as a calcium
or a magnesium salt, a salt of organic bases such as
dicyclohexylamine, N-methyl-D-glucamine,
tris(hydroxymethyl)methylamine, C.sub.1-C.sub.7 alkylamine,
cyclohexylamine, triethanolamine, ethylenediamine, and salts with
amino acids such as arginine, lysine, and the like. In some
embodiments, the therapeutic agent can have a low aqueous
solubility in neutral form. In some embodiments, the
pharmaceutically acceptable salt of a therapeutic agent can have
higher aqueous solubility in comparison to a therapeutic agent in
neutral form.
[0115] Many and varied therapeutic agents can be incorporated by
encapsulation within the synthetic membrane vesicles. A
non-limiting list of therapeutic agent classes include, but are not
limited to, antianginas, antiarrhythmics, antiasthmatic agents,
antibiotics, antidiabetics, antifungals, antihistamines,
antihypertensives, antiparasitics, antineoplastics, antiobesity
agents, antiviral agents, otologicals, cardiac glycosides,
hormones, immunomodulators, monoclonal antibodies,
neurotransmitters, sedatives, vaccines, vasopressors, anesthetics,
amide anaesthetics, corticosteroids, tricyclic antidepressants,
tetracyclic antidepressants, selective serotonin reuptake
inhibitors, steroid receptor modulators, antipsychotic drugs,
antiprotozoal drugs, opioids, antiproliferative agents,
salicylanilides, antihelminthic drugs, vinca alkaloids,
anti-inflammatory agents, anti-depressants, prostaglandins,
phosphodiesterase IV inhibitors; retinoids, steroids,
.beta.-adrenergic receptor ligands, anti-mitotic agents,
microtubule inhibitors, microtubule-stabilizing agents, serotonin
norepinephrine reuptake inhibitors, noradrenaline reuptake
inhibitors, non-steroidal immunophilin-dependent
immunosuppressants, non-steroidal immunophilin-dependent
immunosuppressant enhancers; antimalarial agents, analgesics,
immunosuppressants, expectorants, sulfa drugs, cardiovascular
drugs, central nervous system (CNS) depressants, H2-blockers,
anti-platelet drugs, anticonvulsants, alpha blockers,
beta-blockers, cholinesterase inhibitors, calcium channel blockers,
H1-receptor antagonists, and proteinaceous materials. The
therapeutic agents listed herein can be used in the preparation of
medicaments for the treatment of a disease for which the
therapeutic agent is known to those of skill in the art to be
effective. Therapeutic agents, and diseases for which the
therapeutic agent is effective, can be identified by reference to,
for example, The Physician's Desk Reference, which is incorporated
herein by reference in its entirety.
[0116] Examples of proteinaceous materials that can be incorporated
into the synthetic membrane vesicles, include but are not limited
to, DNA, RNA, proteins of various types, protein hormones produced
by recombinant DNA technology effective in humans, hematopoietic
growth factors, monokines, lymphokines, tumor necrosis factor,
inhibin, tumor growth factor alpha and beta, Mullerian inhibitory
substance, nerve growth factor, fibroblast growth factor,
platelet-derived growth factor, pituitary and hypophyseal hormones
including LH and other releasing hormones.
[0117] Examples of antiarrhythmics, include but are not limited to,
quinidine, procainamide, disopyramide, ajmaline, lidocaine,
tocamide, mexiletine, flecamide, propafenone, moricizine,
propranolol, esmolol, timolol, metoprolol, atenolol, amiodarone,
sotalol, ibutilide, dofetilide, verapamil, diltiazem, and
digoxin.
[0118] Examples of antiasthmatic agents, include but are not
limited to, salbutamol, levalbuterol, terbutaline, bitolterol,
epinephrine, ipratropium bromide, salmeterol, formoterol,
bambuterol, and albuterol.
[0119] Examples of antibiotics, include but are not limited to,
amikacin, gentamicin, kanamycin, neomycin, netilmicin,
streptomycin, tobramycin, paromomycin, geldanamycin, herbimycin,
loracarbef, ertapenem, doripenem, imipenem, meropenem, cefadroxil,
cefazolin, cefalotin, cefalexin, cefaclor, cefamandole, cefoxitin,
cefprozil, cefuroxime, cefixime, cefdinir, cefditoren,
cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten,
ceftizoxime, ceftriaxone, cefepime, ceftobiprole, teicoplanin,
vancomycin, azithromycin, clarithromycin, dirithromycin,
erythromycin, roxithromycin, troleandomycin, telithromycin,
spectinomycin, aztreonam, amoxicillin, ampicillin, azlocillin,
carbenicillin, cloxacillin, dicloxacillin, flucloxacillin,
mezlocillin, meticillin, nafcillin, oxacillin, penicillin,
piperacillin, ticarcillin, bacitracin, colistin, polymyxin b,
ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin,
moxifloxacin, norfloxacin, ofloxacin, trovafloxacin, grepafloxacin,
sparfloxacin, mafenide, prontosil, sulfacetamide, sulfamethizole,
sulfanilimide, sulfasalazine, sulfisoxazole, trimethoprim,
trimethoprim-sulfamethoxazole, demeclocycline, doxycycline,
minocycline, oxytetracycline, tetracycline, arsphenamine,
chloramphenicol, clindamycin, lincomycin, ethambutol, fosfomycin,
fusidic acid, furazolidone, isoniazid, linezolid, metronidazole,
mupirocin, nitrofurantoin, platensimycin, pyrazinamide,
quinupristin, rifampin, thiamphenicol, and timidazole.
[0120] Examples of antidiabetics, include but are not limited to,
tolbutamide, acetohexamide, tolazamide, chlorpropamide, glipizide,
glyburide, glimepiride, gliclazide, repaglinide, nateglinide,
metformin, rosiglitazone, pioglitazone, troglitazone, miglitol,
acarbose, exenatide, liraglutide, taspoglatide, vildagliptin,
sitagliptin, GLP-1, and analog to GLP-1.
[0121] Examples of antifungals, include but are not limited to,
natamycin, rimocidin, filipin, nystatin, amphotericin B, candicin,
miconazole, ketoconazole, clotrimazole, econazole, bifonazole,
butoconazole, fenticonazole, isoconazole, oxiconazole,
sertaconazole, sulconazole, tioconazole, fluconazole, itraconazole,
isavuconazole, ravuconazole, posaconazole, voriconazole,
terconazole, abafungin, terbinafine, amorolfine, naftifine,
butenafine, anidulafungin, caspofungin, micafungin, ciclopirox,
tolnaftate, undecylenic acid, 5-fluorocytosine, and
griseofulvin.
[0122] Examples of antihistamines, include but are not limited to,
aceprometazine, alimemazine, astemizole, azatadine, azelastine,
benadryl, bepotastine, bisulepine, brompheniramine, chlorcyclizine,
chloropyramine, chlorothen, chlorphenamine, cinnarizine,
clemastine, clemizole, clobenzepam, clobenztropine, clocinizine,
cyclizine, cyproheptadine, dacemazine, dexbrompheniramine,
dexchlorpheniramine, diphenhydramine, doxylamine, drixoral,
ebastine, embramine, emedastine, epinastine, etymemazine,
fexofenadine, homochlorcyclizine, hydroxyzine, iproheptine,
isopromethazine, ketotifen, levocabastine, mebhydrolin, mepyramine,
methafurylene, methapyrilene, methdilazine, moxastine,
p-methyldiphenhydramine, pemirolast, pheniramine, phenyltoloxamine,
resporal, rondec, semprex-d, setastine, sominex, talastine,
terfenadine, thenyldiamine, thiazinamium, and triprolidine.
[0123] Examples of antihypertensives, include but are not limited
to, bumetanide, ethacrynic acid, furosemide, torsemidet, epitizide,
hydrochlorothiazide, chlorothiazide, bendroflumethiazide,
indapamide, chlorthalidone, metolazone, amiloride, triamterene,
spironolactone, atenolol, metoprolol, nadolol, oxprenolol,
pindolol, propranolol, timolol, doxazosin, phentolamine, indoramin,
phenoxybenzamine, prazosin, terazosin, tolazoline, bucindolol,
carvedilol, labetalol, clonidine, methyldopa, guanfacine,
amlodipine, felodipine, isradipine, lercanidipine, nicardipine,
nifedipine, nimodipine, nitrendipine, diltiazem, verapamil,
captopril, enalapril, fosinopril, lisinopril, perindopril,
quinapril, ramipril, trandolapril, benazepril, candesartan,
eprosartan, irbesartan, losartan, olmesartan, telmisartan,
valsartan, eplerenone, spironolactone, sodium nitroprus side,
clonidine, guanabenz, methyldopa, moxonidine, guanethidine, and
reserpine.
[0124] Examples of antiparasitics, include but are not limited to,
mebendazole, pyrantel pamoate, thiabendazole, diethycarbazine,
niclosamide, praziquantel, rifampin, amphotericin B, and
melarsoprol.
[0125] Examples of antineoplastics, include but are not limited to,
aclarubicin, altretamine, aminopterin, amrubicin, azacitidine,
azathioprine, belotecan, busulfan, camptothecin, capecitabine,
carboplatin, carmofur, carmustine, chlorambucil, cisplatin,
cladribine, clofarabine, cyclophosphamide, cytarabine,
daunorubicin, decitabine, doxorubicin, epirubicin, etoposide,
floxuridine, fludarabine, 5-fluorouracil, fluorouracil,
gemcitabine, idarubicin, ifosfamide, irinotecan, mechlorethamine,
melphalan, mercaptopurine, methotrexate, mitoxantrone, nedaplatin,
oxaliplatin, pemetrexed, pentostatin, pirarubicin, pixantrone,
procarbazine, pyrimethamine raltitrexed, rubitecan, satraplatin,
streptozocin, thioguanine, triplatin tetranitrate, teniposide,
topotecan, tegafur, trimethoprim, uramustine, vairubicin,
vinblastine, vincristine, vindesine, vinflunine, vinorelbine, and
zorubicin.
[0126] Examples of antiviral agents, include but are not limited
to, abacavir, aciclovir, acyclovir, adefovir, amantadine,
amprenavir, arbidol, atazanavir, atripla, boceprevir, cidofovir,
combivir, darunavir, delavirdine, didanosine, edoxudine, efavirenz,
emtricitabine, enfuvirtide, entecavir, famciclovir, fomivirsen,
fosamprenavir, foscarnet, fosfonet, ganciclovir, ibacitabine,
immunovir, idoxuridine, imiquimod, indinavir, inosine, lamivudine,
lopinavir, loviride, maraviroc, moroxydine, nelfinavir, nevirapine,
nexavir, oseltamivir, penciclovir, peramivir, pleconaril,
raltegravir, ribavirin, rimantadine, ritonavir, saquinavir,
stavudine, tenofovir, tenofovir disoproxil, tipranavir,
trifluridine, trizivir, tromantadine, valaciclovir, valganciclovir,
vicriviroc, vidarabine, viramidine, zalcitabine, zanamivir, and
zidovudine.
[0127] Examples of otologicals, include but are not limited to,
betamethasone, chloramphenicol, chlorhexidine, clioquinol,
dexamethasone, gentamicin, hydrocortisone, lidocaine, miconazole,
neomycin, nitrofural, polymyxin b, prednisolone, rifamycin, and
tetracycline.
[0128] Examples of cardiac glycosides, include but are not limited
to, digitoxin, digoxin, and deslanoside.
[0129] Examples of hormones, include but are not limited to,
adiponectin, adrenocorticotropic hormone, aldosterone,
androstenedione, angiotensinogen, angiotensin, antidiuretic
hormone, antimullerian hormone, atrial-natriuretic peptide, brain
natriuretic peptide, 25-hydroxyvitamin D.sub.3, calcitonin,
1,25-dihydroxyvitamin D.sub.3, cholecystokinin,
corticotropin-releasing hormone, cortisol, dehydroepiandrosterone,
dihydrotestosterone, dopamine, endothelin, enkephalin, epinephrine,
erythropoietin, estradiol, estriol, estrone, follicle-stimulating
hormone, gastrin, ghrelin, glucagon, gonadotropin-releasing
hormone, growth hormone, growth hormone-releasing hormone,
histamine, human chorionic gonadotropin, human placental lactogen,
inhibin, insulin, insulin-like growth factor, leptin, leukotrienes,
lipotropin, luteinizing hormone, melanocyte stimulating hormone,
melatonin, neuropeptide y, norepinephrine, orexin, oxytocin,
pancreatic polypeptide, parathyroid hormone, progesterone,
prolactin, prolactin releasing hormone, prostacyclin,
prostaglandins, relaxin, renin, secretin, serotonin, somatostatin,
testosterone, thrombopoietin, thromboxane, thyroid-stimulating
hormone, thyrotropin-releasing hormone, thyroxine, and
triiodothyronine.
[0130] Examples of immunomodulators, include but are not limited
to, abatacept, abetimus, adalimumab, afelimomab, aflibercept,
afutuzumab, alefacept, anakinra, aselizumab, atlizumab,
atorolimumab, azathioprine, basiliximab, belatacept, belimumab,
bertilimumab, cedelizumab, clenoliximab, certolizumab pegol,
ciclosporin, daclizumab, deforolimus, dorlimomab aritox,
dorlixizumab, efalizumab, erlizumab, elsilimomab, etanercept,
everolimus, faralimomab, fontolizumab, galiximab, gantenerumab,
gavilimomab golimumab, gomiliximab, gusperimus, infliximab,
inolimomab, ipilimumab keliximab, lebrilizumab, leflunomide,
lenalidomide, lerdelimumab, lumiliximab, maslimomab, mepolizumab,
metelimumab, methotrexate, morolimumab, muromonab-cd3, mycophenolic
acid, natalizumab, nerelimomab, ocrelizumab, odulimomab,
omalizumab, otelixizumab, pascolizumab, pexelizumab, pimecrolimus,
reslizumab, rilonacept, rovelizumab, ruplizumab, siplizumab,
sirolimus, tacrolimus, talizumab, telimomab aritox, temsirolimus,
teneliximab, teplizumab, teriflunomide, thalidomide, tocilizumab,
toralizumab, tremelimumab, ustekinumab, vapaliximab, vepalimomab,
visilizumab, zanolimumab, ziralimumab, zolimomab aritox,
zotarolimus, and tetrachlorodecaoxide.
[0131] Examples of monoclonal antibodies, include but are not
limited to, abagovomab, abatacept, abciximab, adalimumab,
adecatumumab, aflibercept, afutuzumab, alacizumab pegol,
alemtuzumab, altumomab, afelimomab, anatumomab mafenatox,
anrukinzumab, apolizumab, arcitumomab, aselizumab, atlizumab,
atorolimumab, bapineuzumab, basiliximab, bavituximab, bectumomab,
belatacept, belimumab, bertilimumab, besilesomab, bevacizumab,
biciromab brallobarbital, bivatuzumab mertansine, blinatumomab,
briakinumab, canakinumab, cantuzumab mertansine, capromab
pendetide, catumaxomab, cedelizumab, certolizumab pegol, cetuximab,
citatuzumab bogatox, cixutumumab, clenoliximab, golimumab,
ustekinumab, conatumumab, dacetuzumab, dacliximab, daclizumab,
denosumab, detumomab, dorlimomab aritox, dorlixizumab, ecromeximab,
eculizumab, edobacomab, edrecolomab, efalizumab, efungumab,
elsilimomab, enlimomab pegol, epitumomab cituxetan, epratuzumab,
erlizumab, ertumaxomab, etanercept, etaracizumab, exbivirumab,
fanolesomab, faralimomab, felvizumab, fezakinumab, figitumumab,
fontolizumab, foravirumab, galiximab, gantenerumab, gavilimomab,
gemtuzumab ozogamicin, golimumab, gomiliximab, ibalizumab,
ibritumomab tiuxetan, igovomab, imciromab, infliximab, intetumumab,
inolimomab, inotuzumab ozogamicin, ibalizumab, ipilimumab,
iratumumab, keliximab, labetuzumab, lemalesomab, lebrilizumab,
lerdelimumab, lexatumumab, libivirumab, lintuzumab, lucatumumab,
lumiliximab, mapatumumab, maslimomab, matuzumab, mepolizumab,
metelimumab, milatuzumab, minretumomab, mitumomab, morolimumab,
motavizumab, muromonab, stamulumab, nacolomab tafenatox, naptumomab
estafenatox, natalizumab, nebacumab, necitumumab, nerelimomab,
nimotuzumab, nofetumomab merpentan, ocrelizumab, odulimomab,
ofatumumab, omalizumab, oportuzumab monatox, oregovomab,
otelixizumab, pagibaximab, palivizumab, panitumumab, panobacumab,
pascolizumab, pemtumomab, pertuzumab, pexelizumab, pintumomab,
priliximab, pritumumab, rafivirumab, ramucirumab, ranibizumab,
raxibacumab, regavirumab, reslizumab, rilonacept, rilotumumab,
rituximab, robatumumab, rovelizumab, rozrolimupab, ruplizumab,
satumomab, sevirumab, sibrotuzumab, siltuximab, siplizumab,
solanezumab, sonepcizumab, sontuzumab, stamulumab, sulesomab,
tacatuzumab tetraxetan, tadocizumab, talizumab, tanezumab,
taplitumomab paptox, tefibazumab, telimomab aritox, tenatumomab,
teneliximab, teplizumab, ticilimumab, tigatuzumab, tocilizumab,
toralizumab, tositumomab, trastuzumab, tremelimumab, tucotuzumab
celmoleukin, tuvirumab, urtoxazumab, ustekinumab, vapaliximab,
vedolizumab, veltuzumab, vepalimomab, visilizumab, volociximab,
votumumab, zalutumumab, zanolimumab, ziralimumab, and zolimomab
aritox.
[0132] Examples of neurotransmitters, include but are not limited
to, acetylcholine, adenosine, adenosine-5'-triphosphate, aspartate,
norepinephrine, dopamine, glycine, serotonin, melatonin, histamine,
glutamate, gamma aminobutyric acid, and
guanosine-5'-triphosphate.
[0133] Examples of sedatives, include but are not limited to,
alprazolam, amobarbital, carisoprodol, chlordiazepoxide,
clomethiazole, clonazepam, diazepam, diphenhydramine, estazolam,
eszopiclone, ethchlorvynol, flunitrazepam, gamma-hydroxybutyrate,
glutethimide, ketamine, lorazepam, methaqualone, methyprylon,
midazolam, nitrazepam, oxazepam, pentobarbital,
phenobarbitoltriazolam, ramelteon, secobarbital, temazepam,
thalidomide, zaleplon, zolpidem, and zopiclone.
[0134] Examples of vaccines, include but are not limited to,
measles vaccine, mumps vaccine, rubella vaccine, varicella vaccine,
inactivated polio vaccine, inactivated influenza vaccine, influenza
a virus subtype H1N1 vaccine, diphtheria toxoid vaccine, tetanus
toxoid vaccine, haemophilus influenzae type B vaccine, hepatitis B
vaccine, hepatitis A vaccine, and pneumoccocal conjugate
vaccine.
[0135] Examples of vasopressors, include but are not limited to,
epinephrine, phenylephrine, dobutamine, isoproterenol,
norepinephrine, aceprometazine, alimemazine, astemizole, azatadine,
azelastine, benadryl, bepotastine, bisulepine, brompheniramine,
chlorcyclizine, chloropyramine, chlorothen, chlorphenamine,
cinnarizine, clemastine, clemizole, clobenzepam, clobenztropine,
clocinizine, cyclizine, cyproheptadine, dacemazine,
dexbrompheniramine, dexchlorpheniramine, diphenhydramine,
doxylamine, drixoral, ebastine, embramine, emedastine, epinastine,
etymemazine, fexofenadine, homochlorcyclizine, hydroxyzine,
iproheptine, isopromethazine, ketotifen, levocabastine,
mebhydrolin, mepyramine, methafurylene, methapyrilene,
methdilazine, moxastine, p-methyldiphenhydramine, pemirolast,
pheniramine, phenyltoloxamine, resporal, rondec, semprex-d,
setastine, sominex, talastine, terfenadine, thenyldiamine,
thiazinamium, and triprolidine.
[0136] Examples of anesthetics, include but are not limited to,
propofol, etomidate, methohexital and sodium thiopental, midazolam,
diazepam, and ketamine, benzocaine, chloroprocaine, cocaine,
cyclomethycaine, dimethocaine, propoxycaine, procaine,
proparacaine, tetracaine, articaine, bupivacaine, carticaine,
dibucaine, etidocaine, levobupivacaine, lidocaine, mepivacaine,
piperocaine, prilocalne, ropivacaine, trimecaine, saxitoxin, and
tetrodotoxin.
[0137] Examples of amide anesthetics, include but are not limited
to, articaine, bupivacaine, carticaine, dibucaine, etidocaine,
levobupivacaine, lidocaine, mepivacaine, piperocaine, prilocalne,
ropivacaine, and trimecaine.
[0138] Examples of corticosteroids, include but are not limited to,
hydrocortisone, hydrocortisone acetate, cortisone acetate,
tixocortol pivalate, prednisolone, methylprednisolone, prednisone,
triamcinolone acetonide, triamcinolone alcohol, mometasone,
amcinonide, budesonide, desonide, fluocinonide, fluocinolone
acetonide, halcinonide, betamethasone, betamethasone sodium
phosphate, dexamethasone, dexamethasone sodium phosphate,
fluocortolone, hydrocortisone-17-butyrate,
hydrocortisone-17-valerate, aclometasone dipropionate,
betamethasone valerate, betamethasone dipropionate, prednicarbate,
clobetasone-17-butyrate, clobetasol-17-propionate, fluocortolone
caproate, fluocortolone pivalate, and fluprednidene acetate.
[0139] Examples of tricyclic antidepressants, include but are not
limited to, amitriptyline, butriptyline, clomipramine, dosulepin,
doxepin, imipramine, lofepramine, trimipramine, desipramine,
nortriptyline, and protriptyline.
[0140] Examples of tetracyclic antidepressants, include but are not
limited to, amoxapine, maprotiline, mianserin, mirtazapine, and
setiptiline.
[0141] Examples of selective serotonin reuptake inhibitors, include
but are not limited to, citalopram, dapoxetine, escitalopram,
fluoxetine, fluvoxamine, paroxetine, sertraline, vilazodone, and
zimelidine.
[0142] Examples of antipsychotic drugs, include but are not limited
to, haloperidol, droperidol, chlorpromazine, fluphenazine,
perphenazine, prochlorperazine, thioridazine, trifluoperazine,
mesoridazine, periciazine, promazine, triflupromazine,
levomepromazine, promethazine, pimozide, chlorprothixene,
flupenthixol, thiothixene, zuclopenthixol, clozapine, olanzapine,
risperidone, quetiapine, ziprasidone, amisulpride, asenapine,
paliperidone, aripiprazole, and bifeprunox.
[0143] Examples of antiprotozoal drugs, include but are not limited
to, eflornithine, furazolidone, melarsoprol, metronidazole,
ornidazole, paromomycin sulfate, pentamidine, pyrimethamine, and
timidazole.
[0144] Examples of opioids, include but are not limited to,
endorphins, enkephalins, dynorphins, endomorphins, codeine,
morphine, thebaine, oripavine, diacetylmorphine, dihydrocodeine,
hydrocodone, hydromorphone, nicomorphine, oxycodone, oxymorphone,
fentanyl, alphamethylfentanyl, alfentanil, sufentanil,
remifentanil, carfentanyl, ohmefentanyl, pethidine, ketobemidone,
allylprodine, prodine, propoxyphene, dextropropoxyphene,
dextromoramide, bezitramide, piritramide, methadone, dipipanone,
levomethadyl acetate, loperamide, diphenoxylate, dezocine,
pentazocine, phenazocine, buprenorphine, dihydroetorphine,
etorphine, butorphanol, nalbuphine, levorphanol, levomethorphan,
lefetamine, meptazinol, tilidine, tramadol, tapentadol, nalmefene,
naloxone, and naltrexone.
[0145] Examples of antiproliferative agents, include but are not
limited to, aclarubicin, altretamine, aminopterin, amrubicin,
azacitidine, azathioprine, belotecan, busulfan, camptothecin,
capecitabine, carboplatin, carmofur, carmustine, chlorambucil,
cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine,
daunorubicin, decitabine, docetaxel, doxorubicin, epirubicin,
etoposide, floxuridine, fludarabine, 5-fluorouracil, fluorouracil,
gemcitabine, idarubicin, ifosfamide, irinotecan, mechlorethamine,
melphalan, mercaptopurine, methotrexate, mitoxantrone, nedaplatin,
oxaliplatin, paclitaxel, pemetrexed, pentostatin, pirarubicin,
pixantrone, procarbazine, pyrimethamine raltitrexed, rubitecan,
satraplatin, sirolimus, streptozocin, thioguanine, triplatin
tetranitrate, teniposide, topotecan, tegafur, trimethoprim,
uramustine, valrubicin, vinblastine, vincristine, vindesine,
vinflunine, vinorelbine, and zorubicin.
[0146] Examples of salicylanilides, include but are not limited to,
niclosamide, oxyclozanide, and rafoxanide.
[0147] Examples of antihelminthic drugs, include but are not
limited to, abamectin, albendazole, diethylcarbamazine,
mebendazole, niclosamide, ivermectin, suramin, thiabendazole,
pyrantel pamoate, levamisole, praziquantel, triclabendazole,
flubendazole, fenbendazole, emodepside, and monepantel.
[0148] Examples of vinca alkaloids, include but are not limited to,
vinblastine, vincristine, vindesine and vinorelbine.
[0149] Examples of anti-inflammatory agents, include but are not
limited to, phenylbutazone, mofebutazone, oxyphenbutazone,
clofezone, kebuzone, indometacin, sulindac, tolmetin, zomepirac,
diclofenac, alclofenac, bumadizone, etodolac, lonazolac, fentiazac,
acemetacin, difenpiramide, oxametacin, proglumetacin, ketorolac,
aceclofenac, bufexamac, piroxicam, tenoxicam, droxicam, lornoxicam,
meloxicam, ibuprofen, naproxen, ketoprofen, fenoprofen, fenbufen,
benoxaprofen, suprofen, pirprofen, flurbiprofen, indoprofen,
tiaprofenic acid, oxaprozin, ibuproxam, dexibuprofen,
flunoxaprofen, alminoprofen, dexketoprofen, mefenamic acid,
tolfenamic acid, flufenamic acid, meclofenamic acid, celecoxib,
rofecoxib, valdecoxib, parecoxib, etoricoxib, lumiracoxib,
nabumetone, niflumic acid, azapropazone, glucosamine, benzydamine,
glucosaminoglycan polysulfate, proquazone, orgotein, nimesulide,
feprazone, diacerein, morniflumate, tenidap, oxaceprol, and
chondroitin sulfate.
[0150] Examples of cancers that can be treated with an anticancer
agent include, but are not limited to, head and neck cancer, breast
cancer, colorectal cancer, gastric cancer, hepatic cancer, bladder
cancer, cervical cancer, endometrial cancer, lung cancer (non-small
cell), ovarian cancer, pancreatic cancer, prostate cancer;
choriocarcinoma (lung cancer); hairy cell leukemia, chronic
lymphotic leukemia, acute lymphocytic leukemia (breast &
bladder), acute myelogenous leukemia, Hodgkin's lymphoma,
non-Hodgkin's lymphoma (osteogenic sarcoma, adult soft tissue
sarcoma), meningeal leukemia, multiple myeloma, chronic myelogenous
leukemia, erythroleukemia, and T-cell lymphoma.
[0151] Examples of inflammatory and autimmune diseases that can be
treated with an inflammatory agent include, but are not limited to,
B cell disorders, T cell disorders, rheumatoid arthritis (RA),
systemic lupus erythematosus (SLE), Sjogren's syndrome, immune
thrombocytopenic purpura (ITP), multiple sclerosis (MS), myasthenia
Gravis (MG), Graves disease, psoriasis, Hashimoto's disease, immune
thrombocytopenic purpura, scleroderma, and inflamatory bowel
disease (e.g. Crohn's disease and ulcerative colitis).
[0152] In some embodiments, the therapeutic agent can be used
singly or in combination with the limitation that the amount of the
physiologically active substance in the pharmaceutical composition
be sufficient to enable the diagnosis of, prophylaxis against, or
treatment of an undesired condition in a living being. In some
embodiments, the pharmaceutical compositions can be administered to
a living being by any desired route, for example, intramuscular,
intra articular, epidural, intraperitoneal, subcutaneous, intra
lymphatic, oral, submucosal, transdermal, rectal, vaginal,
intranasal, intraocular, and by implantation under different kinds
of epithelia, including the bronchial epithelia, the
gastrointestinal epithelia, the urogenital epithelia, and the
various mucous membranes of the body. Generally, the dosage will
vary with the age, condition, sex and extent of the undesired
condition in the patient, and can be determined by one skilled in
the art. In some embodiments, the dosage range appropriate for
human use includes a range of from 0.1 to 6,000 mg of the
therapeutic agent per square meter of surface area. Alternate
dosage range can be based on weight instead of surface area. In one
embodiment, a human dosage of bupivacaine can be 50-1,000 mg,
100-600 mg 100-350 mg. For example, the human dosage of bupivacaine
can be approximately 300 mg.
Methods of Preparation
[0153] Some embodiments relate to a process for preparing a large
diameter synthetic membrane vesicle(s) composition comprising the
steps of, forming a first component by mixing a first aqueous phase
and an organic phase, said organic phase comprising an organic
solvent, at least one amphipathic lipid, and at least one neutral
lipid, encapsulating said first component in a second aqueous phase
to provide a second component using an atomizing nozzle as
disclosed and described herein, said second component comprising an
aqueous phase, removing the organic solvent from the second
component to form a composition of large diameter synthetic
membrane vesicle particles, wherein the removing can be
accomplished by contacting the second component with a gas,
optionally heating and optionally filtering the composition by
particle concentration. Such steps may be combined with other
steps. In some embodiments the lipid phase can include cholesterol.
In some embodiments, the organic solvent can be a volatile
water-immiscible or sparingly miscible solvent. In some
embodiments, the first component can be a first emulsion. In some
embodiments, the second component can be a second emulsion. In some
embodiments, the second component can be a droplet. In some
embodiments, the large diameter synthetic membrane vesicle(s) can
be multivesicular liposomes.
[0154] Some embodiments relate to a process for preparing a large
diameter synthetic membrane vesicle(s) composition comprising the
steps of, forming a first component by mixing a first aqueous phase
and an organic phase, said organic phase comprising an organic
solvent, at least one amphipathic lipid, and at least one neutral
lipid, encapsulating said first component in a second aqueous phase
to provide a second component, said second component comprising an
aqueous phase, removing the organic solvent from the second
component to form a composition of large diameter synthetic
membrane vesicle particles, wherein the removing can be
accomplished by using a solvent removal chamber as disclosed and
described herein, and optionally filtering the composition by
particle concentration. Such steps may be combined with other
steps. In some embodiments the lipid phase can include cholesterol.
In some embodiments, the organic solvent can be a volatile
water-immiscible or sparingly miscible solvent. In some
embodiments, the first component can be a first emulsion. In some
embodiments, the second component can be a second emulsion. In some
embodiments, the second component can be a droplet. In some
embodiments, the large diameter synthetic membrane vesicle(s) can
be multivesicular liposomes.
[0155] Some embodiments relate to a process for preparing a large
diameter synthetic membrane vesicle(s) composition comprising the
steps of, forming a first component by mixing a first aqueous phase
and an organic phase, said organic phase comprising an organic
solvent, at least one amphipathic lipid, and at least one neutral
lipid, encapsulating said first component in a second aqueous phase
to provide a second component using an atomizing nozzle as
disclosed and described herein, said second component comprising an
aqueous phase, removing the organic solvent from the second
component to form a composition of large diameter synthetic
membrane vesicle particles, wherein the removing can be
accomplished by using a solvent removal chamber as disclosed and
described herein, and optionally filtering the composition by
particle concentration. Such steps may be combined with other
steps. In some embodiments the lipid phase can include cholesterol.
In some embodiments, the organic solvent can be a volatile
water-immiscible or sparingly miscible solvent. In some
embodiments, the first component can be a first emulsion. In some
embodiments, the second component can be a second emulsion. In some
embodiments, the second component can be a droplet. In some
embodiments, the large diameter synthetic membrane vesicle(s) can
be multivesicular liposomes.
[0156] Some embodiments relate to a process for preparing a
multivesicular liposome composition comprising the steps of,
forming a first component by mixing a first aqueous phase and an
organic phase, said organic phase comprising a volatile
water-immiscible or sparingly miscible solvent, at least one
amphipathic lipid, and at least one neutral lipid, encapsulating
said first component in a second aqueous phase to provide a second
component, said second component comprising an aqueous phase,
removing the volatile water-immiscible or sparingly miscible
solvent from the second component to form a composition of MVL
particles, wherein the removing can be accomplished by contacting
the second component with a gas, and optionally filtering the
multivesicular liposome composition by particle concentration. Such
steps may be combined with other steps. In some embodiments the
lipid phase can include cholesterol.
[0157] First Component
[0158] In embodiments that include a first component, the first
component can be formed by mixing two phases, such as an organic
phase and a first aqueous phase. In some embodiments, a therapeutic
agent can be added to the organic phase. In some embodiments, a
therapeutic agent can be added to the first aqueous phase. In some
embodiments, a therapeutic agent can be added to both the organic
phase and the first aqueous phase. In some embodiments, the organic
phase can include at least one amphipathic lipid, at least one
neutral lipid, and an organic solvent. In some embodiments, the
therapeutic agent can be in the form of a pharmaceutically
acceptable salt.
[0159] In some embodiments, the mixing of the two phases can be
accomplished using ultrasound. In some embodiments, the mixing of
the two phases can be accomplished using high pressure
emulsification. Such emulsification utilizes an atomizing nozzle as
disclosed and described herein. In some embodiments, the mixing of
the two phases can be accomplished using mechanical processes
including using high-shear type devices, rotor/stator and
homogenizers, shear-type mixer, static mixer, impeller, porous
pipe, any of the disclosed mechanical processes optionally in
combination with a heat exchanger, or other processes known to
produce water-in-oil emulsions. In some embodiments, the mixing of
the two phases can be accomplished using a combination of
ultrasound and high pressure emulsification. In some embodiments,
the mixing of the two phases can be accomplished using a
combination of mechanical processes performed by a device selected
from the group consisting of high-shear type devices, rotor/stator
mixers and homogenizers, shear-type mixer, static mixer, impeller,
porous pipe, high energy vibration, injection into a high velocity
liquid stream such as in an aspirator, and the like.
[0160] In some embodiments, the first component can comprise
particles having an average diameter in the range from about 0.1
.mu.m to about 100 .mu.m. For example, the particles can have an
average diameter of at least about 0.1 .mu.m, 1 .mu.m, 5 .mu.m, 10
.mu.m, 15 .mu.m, 20 .mu.m, 50 .mu.m, or 100 .mu.m, or a diameter
within a range defined by any of two of the preceding values. In
some embodiments, the particles can have an average diameter in the
range from about 0.2 .mu.m to about 100 .mu.m, from about 0.2 .mu.m
to about 50 .mu.m, from about 0.5 .mu.m to about 30 .mu.m, from
about 0.5 .mu.m to about 10 .mu.m, from about 10 .mu.m to about 50
.mu.m, from about 15 .mu.m to about 45 .mu.m, or from about 20
.mu.m to about 40 .mu.m. In a typical embodiment, the particles can
have an average diameter in the range of from about 0.5 .mu.m to
about 3 .mu.m.
[0161] In some embodiments, the first component can be formed at a
temperature in the range from about -5.degree. C. to about
99.degree. C., from about 0.degree. C. to about 60.degree. C., from
about 2.degree. C. to about 40.degree. C., from about 4.degree. C.
to about 20.degree. C., from about 5.degree. C. to about 50.degree.
C., from about 10.degree. C. to about 40.degree. C., or from about
15.degree. C. to about 35.degree. C., or from about 10.degree. C.
to about 20.degree. C.
[0162] In some embodiments, the organic solvent can be selected
from the group consisting of diethyl ether, tert-butylmethyl ether,
tetrahydrofuran, sevoflurane, desflurane, isoflurane, and
enflurane. In some embodiments, the organic solvent can be selected
from the group consisting of halothane, chloroform, and
dichloromethane. In some embodiments, the organic solvent can be
selected from the group consisting of ethyl acetate, hexane,
hexanes, cyclohexane, pentane, cyclopentane, petroleum ether, and
toluene. In some embodiments, the organic solvent can be selected
from the group consisting of freons, chlorofluorocarbons (CFCs) and
hydrochlorofluorocarbons (HCFCs) with boiling points above
15.degree. C. In a typical embodiment, the organic solvent can be
selected from the group consisting of chloroform, and
dichloromethane. For example, the organic solvent can be
dichloromethane.
[0163] In some embodiments, the first aqueous phase can be selected
from the group consisting of water solutions including one or more
components selected from the group consisting of a therapeutic
agent, dextrose, lysine, dextrose/lysine, sodium chloride,
hydrochloric acid, phosphoric acid, an osmotic pressure adjusting
agent such as a sugar, dextrose, sucrose, trehalose, fructose,
sorbitan, glycerol, or manitol, a therapeutic agent solubility
enhancer, and pH modifying agents such as sodium hydroxide,
arginine, histidine, sodium borate, acids, bases,
(hydroxymethyl)aminomethane, or Good's buffers. In some
embodiments, the concentration of any one component in the aqueous
phase, other than the therapeutic agent, can be in the range from
about 1 .mu.M to about 1M, from about 1 mM to about 500 mM, from
about 10 mM to about 400 mM, from about 100 mM to about 300 mM. In
a typical embodiment, any one therapeutic agent component if used
in the first aqueous phase can be in the range from about 1 mM to
about 1M. For example, the therapeutic agent component can be
approximately 200 mM. In a typical embodiment, the concentration of
any one component in the first aqueous phase can be in the range
from about 100 mM to about 300 mM. For example, the concentration
can be 200 mM. In some embodiments, the first aqueous phase can
include phosphoric acid as a component. In some embodiments, the
concentration of phosphoric acid can be in the range from about 1
.mu.M to about 1M, from about 10 .mu.M to about 750 mM, from about
1 mM to about 500 mM or from about 10 mM to about 250 mM. In a
typical embodiment, the concentration of phosphoric acid can be in
the range from about 100 mM to about 300 mM. For example, the
concentration of phosphoric acid can be 200 mM.
[0164] In some embodiments, the organic phase can include an
amphipathic lipid. In some embodiments, the amphipathic lipid can
be selected from the group consisting of soya lecithin,
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3-phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol, and mixture thereof. In
a typical embodiment, the amphipathic lipid can be selected from
the group consisting of 1,2-dierucoyl-sn-glycero-3-phosphocholine
(DEPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) and
mixtures thereof. For example, the amphipathic lipid can be a
mixture of 1,2-dierucoyl-sn-glycero-3-phosphocholine (DEPC) and
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG). In some
embodiments the ratio of 1,2-dierucoyl-sn-glycero-3-phosphocholine
(DEPC) to 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) can
be in the range from about 100:1 to about 1:10, from about 50:1 to
about 1:1, from about 25:1 to about 2:1, from about 15:1 to about
10:1, from about 10:1 to about 30:1, or from about 15:1 to about
20:1. For example, the ratio of
1,2-dierucoyl-sn-glycero-3-phosphocholine (DEPC) to
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) can be about
16.8:1.
[0165] In some embodiments, the organic phase can include a neutral
lipid. In some embodiments, the neutral lipid can be selected from
the group consisting of triolein, tripalmitolein, trimyristolein
trilinolein, tributyrin, tricaprylin, tricaproin, and tricaprin,
and mixture thereof. In a typical embodiment, the neutral lipid can
be selected from the group consisting of tricaprylin, tricaproin,
and tricaprin, and mixtures thereof. For example, the neutral lipid
can be tricaprylin. In some embodiments the ratio of amphipathic
lipid to neutral lipid can be in the range from about 50:1 to about
1:5, from about 25:1 to about 1:2, from about 15:1 to about 1:1,
from about 10:1 to about 2:1 or from about 6:1 to about 3:1. For
example, the ratio of amphipathic lipid to neutral lipid can be
about 4.4:1.
[0166] In some embodiments, the organic phase can include
cholesterol. In some embodiments the ratio of amphipathic lipid to
cholesterol can be in the range from about 50:1 to about 1:10, from
about 25:1 to about 1:5, from about 10:1 to about 1:2, from about
5:1 to about 1:1 or from about 3:1 to about 1.5:1. For example, the
ratio of amphipathic lipid to cholesterol can be about 1.8:1. In
some embodiments the ratio of cholesterol to neutral lipid can be
in the range from about 50:1 to about 1:10, from about 25:1 to
about 1:5, from about 10:1 to about 1:2, from about 5:1 to about
1:1 or from about 3:1 to about 2:1. For example, the ratio of
cholesterol to neutral lipid can be about 2.4:1. In a typical
embodiment, any one therapeutic agent component if used in the
organic phase can be in the range from about 1 mM to about 1M. For
example, the therapeutic agent component can be approximately 200
mM.
[0167] In some embodiments, the aqueous phase or organic phase can
comprise the therapeutic agent. In a typical embodiment, the
therapeutic agent can be selected from the group including
semisynthetic aminoglycoside antibiotics such as amikacin;
antidiabetics; peptides such as insulin; antitumor drugs such as
paclitaxel; antineoplastics including cytarabine, 5-fluorouracil
and floxuridine; alkaloid opiate analgesics including morphine and
hydromorphine; local anesthetics including bupivacaine; synthetic
anti-inflamniatory adrenocortical steroids including dexamethasone;
antimetabolites including methotrexate; glycopeptide antibiotics
including bleomycin; vincaleukoblastines and stathmokinetic
oncolytic agents including vincristine and vinblastine; hormones,
plasma proteins, cytokines, growth factors, DNA and RNA from a
variety of organisms, and antisense oligonucleotides. In some
embodiments, the therapeutic agent can be an amide anesthetic. In
some embodiments, the therapeutic agent can be selected from the
group consisting of bupivacaine, mepivacaine, ropivacaine,
lidocaine, pyrrocaine, prilocalne, their stereoisomers, and
combinations thereof. In a typical embodiment, the therapeutic
agent can be bupivacaine or a therapeutically acceptable salt
thereof. For example, bupivacaine can be a free base. In some
embodiments, the aqueous phase can comprise an acid in sufficient
quantity to maintain the bupivacaine or a therapeutically or
pharmaceutically acceptable salt thereof in the first aqueous
phase.
[0168] In some embodiments, the first component can be formed by
mixing two phases. In some embodiments, the first component can be
an emulsion. In some embodiments, the first component can be in the
form of droplets. In some embodiments, the first component droplets
can be formed by mixing two phases, such as an organic phase and a
first aqueous phase where the speed of mixing can control the size
of the first component droplets. In some embodiments, the size of
the first component droplets can have an average diameter of at
least about 0.1 .mu.m, 1 .mu.m, 5 .mu.m, 10 .mu.m, 15 .mu.m, 20
.mu.m, 50 .mu.m, or 1000 .mu.m, or a diameter within a range
defined by any of two of the preceding values. In some embodiments,
the average first component droplet size can be preferably between
0.5 .mu.m and 2 .mu.m. For example, the first component droplet
size can be approximately 1 .mu.m. In some embodiments, the first
component droplet can be an emulsion droplet.
[0169] Second Component Droplet
[0170] In some embodiments, the first component can then be
combined with a second aqueous phase to provide a second component
droplet. The second component droplet can be formed by combining
the first component with the second aqueous phase using a
three-fluid nozzle to form a first component/second aqueous phase
mixture. In some embodiments, the first fluid applied to the nozzle
is a first liquid, made up of a first component, the second fluid
applied to the nozzle is a second liquid made up of a second
aqueous phase, and the third fluid can be considered to be a gas
(or a gas/vapor mixture), such as nitrogen gas (or a nitrogen
gas/aqueous vapor mixture) or air scrubbed of CO.sub.2, or air free
of, or substantially free of CO.sub.2. In some embodiments,
contacting the first component/second aqueous phase mixture with
the third fluid creates the second component droplets as the gas
acts to shear the first emulstion/second aqueous phase mixture into
droplets. In some embodiments, the volume:volume ratio of first
component to second aqueous phase can be in the range of from about
1:1000 to about 1000:1, in the range of from about 1:500 to about
500:1, in the range of from about 1:50 to about 50:1, in the range
of from about 1:10 to about 5:1, or in the range of from about 1:5
to about 5:1. In a typical embodiment, the volume:volume ratio of
first component to second aqueous phase can be in the range of from
about 1:3 to about 3:1. Alternately, the volume:volume ratio of
first component to second aqueous phase can be in the range of from
about 2:1 to 1:2. For example, the volume:volume ratio of first
component to second aqueous phase can be about 1:1. In some
embodiments, the first component can be an emulsion.
[0171] In some embodiments, the second aqueous phase can be
selected from the group consisting of water solutions of, a
therapeutic agent, or pharmaceutically acceptable salt thereof,
dextrose, lysine, dextrose/lysine, sodium chloride, hydrochloric
acid, phosphoric acid, an osmotic pressure adjusting agent such as
a sugar, dextrose, sucrose, trehalose, fructose, sorbitan,
glycerol, or manitol, a therapeutic agent solubility enhancer, pH
modifying agents such as sodium hydroxide, arginine, histidine,
sodium borate, acids, bases, (hydroxymethyl)aminomethane, or Good's
buffers, and mixtures thereof. In some embodiments, the
concentration of any one component in the second aqueous phase can
be in the range from about 1 .mu.M to about 1M, from about 10 .mu.M
to about 750 mM, from about 100 .mu.M to about 500 mM, 1 mM to
about 250 mM, from about 10 mM to about 150 mM, from about 50 mM to
about 125 mM, from about 100 mM to about 200 mM, from about 100 mM
to about 500 mM, or from about 200 mM to about 400 mM. In a typical
embodiment, the concentration of any one component in the second
aqueous phase can be in the range from about 50 mM to about 270
mM.
[0172] In some embodiments, the second aqueous phase can be an
aqueous dextrose/lysine solution. In some embodiments, the
concentration of dextrose can be in the range from about 1 mM to
about 500 mM from about 10 mM to about 300 mM, from about 25 mM to
about 200 mM or from about 50 mM to about 150 mM. In a typical
embodiment, the concentration of dextrose can be in the range from
about 60 mM to about 90 mM. For example, the concentration of
dextrose can be about 80 mM. In some embodiments, the concentration
of lysine can be in the range from about 1 mM to about 750 mM, from
about 10 mM to about 500 mM, from about 50 mM to about 400 mM or
from about 100 mM to about 200 mM. In a typical embodiment, the
concentration of lysine can be in the range from about 150 mM to
about 250 mM. For example, the concentration of lysine can be about
200 mM.
[0173] In some embodiments, a first component can be mixed with a
second organic phase comprising one or more second amphipathic
lipids prior to combination with a second aqueous phase to provide
a second component. In some embodiments, the second component can
be in the form of droplets. For example, the second organic phase
can include the same or different amphipathic lipids than in the
first component. The second component droplet can be formed by
combining the first component with the second aqueous phase using a
three fluid nozzle with an additional inlet for the second organic
phase. The second organic phase can alternatively be pumped into
the conduit feeding first component to the three fluid nozzle with
an optional static mixer in the conduit between the point of second
organic phase addition and the three fluid nozzle. Some active
agents can interact with the lipids (e.g. peptides or proteins,
have seen this already, especially the charged lipids like PG, e.g.
DPPG). For this and other reasons e.g. biocompatibility, storage
stability, vibration stability or release rate modification, it may
be desirable to have the outside layer of phospholipids composition
different from the inside chamber wall composition (e.g. PG only on
the outside where it gives charge stability to the MVL but not
present during high shear mixing with the active agent first
aqueous). In some embodiments, the outside layer of phospholipid is
put on the particle when the first component contacts the second
aqueous phase, inside the nozzle and is subsequently atomized and
it comes from the phospholipids left dissolved in the solvent after
the first emulsion is made. Phospholipids with the desired outside
composition, dissolved in solvent, can be injected into the first
fluid conduit, just before the 3 fluid nozzle, and just before an
optional static or other mixer so that these lipids are present and
dissolved in the solvent phase of the first component. These
phospholipids would only contact active containing first component
droplets for seconds under mild shear condition and would thus not
interact appreciably with the outer layer lipids. Some of these
added lipids may get incorporated between the chambers of the
forming MVL which the 4 fluid nozzle avoids.
[0174] In some embodiments, the one or more second amphipathic
lipids in the second organic phase can be selected from the group
consisting of soya lecithin,
1,2-dioleoyl-sn-glycero-3-phosphocholine,
1,2-dilauroyl-sn-glycero-3-phosphocholine,
1,2-dimyristoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphocholine,
1,2-distearoyl-sn-glycero-3-phosphocholine,
1,2-diarachidoyl-sn-glycero-3.about.phosphocholine,
1,2-dibehenoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine,
1,2-dieicosenoyl-sn-glycero-3-phosphocholine,
1,2-dierucoyl-sn-glycero-3-phosphocholine,
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol,
1,2-dioleoyl-sn-glycero-3-phosphoglycerol, and mixture thereof. In
a typical embodiment, the amphipathic lipid can be selected from
the group consisting of 1,2-dierucoyl-sn-glycero-3-phosphocholine
(DEPC), 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) and
mixtures thereof.
Solvent Removal
[0175] In some embodiments, the organic solvent, such as methylene
chloride, can be nearly or completely removed from a droplet by
further contacting the droplet with a gas (or a gas with some
degree of aqueous phase vapor), such as nitrogen gas, or air
scrubbed of CO2, or air substantially free of CO2 in a solvent
removal chamber. As described below, in some embodiments, the gas
can have 50% to 100% relative humidity. For example, the humidity
can be 100%. In some embodiments, the droplet can remain suspended
in the gas within the solvent removal chamber until nearly all of
the organic solvent is removed thereby creating a large diameter
synthetic membrane vesicle particle. In some embodiments, the
solvent removal can be done in a solvent removal chamber as
discussed herein. The large diameter synthetic membrane vesicle
particle can then be collected along with multiple other large
diameter synthetic membrane vesicle particles, forming large
diameter synthetic membrane vesicles suspended in a continuous
aqueous phase. In some embodiments, the large diameter synthetic
membrane vesicles suspended in a continuous aqueous phase can
undergo further processing. In some embodiments, the large diameter
synthetic membrane vesicles can be multivesicular liposomes.
[0176] Particle Concentration
[0177] Optionally, the large diameter synthetic membrane vesicles
suspended in the continuous aqueous solution can be processed to
modify/exchange the continuous aqueous solution via a particle
concentration system. In this document the term concentration unit,
concentration apparatus, concentration system,
particle-concentration system, particle-concentrating device, and
particle concentrator are meant to encompass units and processes
that remove some of the particle suspending medium of a particle
suspension and therefore concentrating the particle concentration
as well as encompassing the exchange of the suspending medium with
a new suspending medium, performed in one step or incrementally.
These two processes are closely related as exchanging the
suspending medium can be accomplished by concentrating the
suspension and adding new suspending medium. These terms relate to
concentrating the particle suspension and exchanging the suspending
medium to be done separately or simultaneously. In some
embodiments, the large diameter synthetic membrane vesicles can be
multivesicular liposomes.
[0178] In one embodiment, the continuous aqueous solution can be
modified/exchanged by diafiltration or cross-flow filtration. The
diafiltration can achieve several objectives, including: exchanging
an aqueous solution by an isotonic solution, concentrating of the
large diameter synthetic membrane vesicles particles, removing
unencapsulated drug, and removing of residual organic solvent. In
some embodiments, the large diameter synthetic membrane vesicles
can be multivesicular liposomes.
[0179] Particle concentration, such as through diafiltration, can
be a method employed for the filtration, purification, and
separation of the large diameter synthetic membrane vesicles
particles from complex mixtures by virtue of the physical
characteristics of the large diameter synthetic membrane vesicles
particles. For example, the most common physical characteristic
used is size. This filtration can involve cross-flow filtration, as
opposed to dead-end filtration. In cross-flow filtration, a
suspension can be circulated under pressure and in contact with a
filter, so that a permeate (the material which passes through the
filter) leaves the system, and a retentate (the material which does
not pass through the filter) is left behind and exits the filter
housing through a port different than the permeate and can be
recirculated through the filter. The suspension then becomes
concentrated in material that does not pass through the filter. In
processes in which a first solution is to be exchanged for a second
solution, the second solution is introduced on the retentate side
of the filter, until the permeate gradually consists of the second
solution. By this time, the first solution has been flushed from
the suspension. An additional consequence of suspending medium
exchange is to further remove any residual organic solvent, such as
methylene chloride which has a small but appreciable water
solubility and is thus removed in the permeate stream. Likewise the
therapeutic agent released during the large diameter synthetic
membrane vesicles formation can also be removed in the permeate. In
some embodiments, the large diameter synthetic membrane vesicles
can be multivesicular liposomes.
[0180] In some embodiments, aseptic methods can be included with
any of the described methods and apparatus described herein.
[0181] Although sterilization of a final filled container as a
dosage form is often a preferred process in pharmaceutical
manufacture for the assurance of minimal risk of microbial
contamination in a lot, the large diameter synthetic membrane
vesicles manufactured in the present processes can be susceptible
to unacceptable damage when subjected to some terminal
sterilization techniques, such as autoclaving and gamma
irradiation. In the absence of validated, non-damaging terminal
sterilization, some embodiments of the invention utilize aseptic
techniques, in which the product is prepared according to a
carefully designed series of aseptic steps. These are designed to
prevent the introduction of viable microorganisms into components,
where sterile, or once an intermediate process has rendered the
bulk product or its components free from viable microorganisms.
Products defined as aseptically processed can consist of components
that have been sterilized by aseptic means. For example, bulk
products which are filterable liquids, can be sterilized by
filtration. Final empty container components can be sterilized by
heat; dry heat for glass vials and autoclaving for rubber seal
components. The requirements for properly designed, validated and
maintained filling and processing facilities are directed to: an
air environment free from viable microorganisms, and designed to
permit effective maintenance of air supply units; training of
personnel who are adequately equipped and gowned. Available
published standards for controlled work areas include: Federal
Standard No. 209B, Clean Room and Work Station Requirements for a
Controlled Environment, Apr. 24, 1973; NASA Standard for Clean Room
and Work Stations for Microbially Controlled Environment,
publication NHB5340.2, August 1967; and Contamination Control of
Aerospace Facilities, U.S. Air Force, T. O. 00-25-203, Dec. 1,
1972, change 1, Oct. 1, 1974. In some embodiments, the large
diameter synthetic membrane vesicles can be multivesicular
liposomes.
[0182] In aseptic processing, one of the most important laboratory
controls is the establishment of an environmental monitoring
program. Samples can be collected from areas in which components
and product are exposed to the environment, including mixing rooms
and component preparation areas. Microbiological quality of aseptic
processing areas is monitored to determine whether or not aseptic
conditions are maintained during filling and closing activities.
Routine sampling and testing of the room air, floors, walls, and
equipment surfaces is carried out. This program establishes the
effectiveness of cleaning and sanitizing equipment and product
contact surfaces, and ensures that potential contaminants are held
to an acceptable level. The disinfectants are checked to assure
that their efficacy against normal microbial flora is maintained.
Sampling schedules, including locations and frequency of sampling,
are maintained. Passive air samplers such as settling plates (Petri
dishes) are employed as well.
[0183] Aseptic assembly operations can be validated by the use of a
microbiological growth nutrient medium to simulate sterile product
filling operations, known as "sterile media fills." The nutrient
medium can be manipulated and exposed to the operators, equipment,
surfaces and environmental conditions to closely simulate the same
exposure which the product itself will undergo. The sealed drug
product containers filled with the media can then be incubated to
detect microbiological growth and the results are assessed to
determine the probability that any given unit of drug product may
become contaminated during actual filling and closing operations.
Media filling, in conjunction with comprehensive environmental
monitoring can be particularly valuable in validating the aseptic
processing of sterile solutions, suspensions, and powders. Filling
liquid media, as part of validating the processing of powders, may
necessitate use of equipment and/or processing steps that would
otherwise not be attendant to routine powder operations.
[0184] In some embodiments, clean-in-place (CIP) and
sterilize-in-place (SIP) procedures can be utilized by methods
generally known in the art. Some embodiments include monitoring of
the temperature at the steam traps. According to this procedure, as
steam is admitted into the vessels and fill lines to effect
sterilization, the temperature at the outlet points is monitored
until bacterial kill is assured. At this point, the seals can be
closed, and the system can be sterilized for further use.
Subsequently, the equipment can be used aseptically in a
non-sterile room environment. The systems described herein, may
also comprise additional components (e.g. valves, steam lines,
condensate drains) to facilitate sterilization by
steaming-in-place.
[0185] Sterility testing of product lots can be carried out
directly after the lot is manufactured as a final product quality
control test. Testing can be done in accordance with various
procedures found in the U.S. Pharmacopeia (U.S. P.) and FDA
regulations.
[0186] Sterile filtration of all fluids which enter the
manufacturing system is essential for an aseptic process, as
envisioned in certain embodiments of the present application.
Rating of pore sizes of filter membranes is by a nominal rating
reflecting the capacity of the membrane to retain microorganisms of
size represented by specified trains, not by determination of an
average pore size and statement of distribution of sizes.
Sterilizing filter membranes include those capable of retaining
100% of a culture of 10.sup.7 organisms of a strain of Pseudomonas
diminua (ATTC 19146) per square cm of membrane surface under a
pressure of not less than 30 psi. Such filter membranes can be
nominally rated 0.22 .mu.m or 0.2 .mu.m, depending on the
manufacturer. Bacterial filter membranes capable of retaining only
larger microorganisms (including Serratia Marcescens (ATTC 14756))
are labeled with a nominal rating of 0.45 .mu.m. Filter membranes
used in the present processes can be of the 0.2 .mu.m type, and can
be used in all lines feeding from liquid solution and gas storage
tanks to vessels and transfer lines used in the methods disclosed
herein.
[0187] In some embodiments the process apparatus is sterilized
before use and isolated from the environment by the use of
sterilizing filters on all apparatus inputs and outputs where the
apparatus includes the sterilized product vessel. In this
embodiment sterile product can be produced in an aseptic fashion,
in a non sterile environment.
[0188] Description of the Instant Devices and their Methods of
Use
[0189] The present embodiments will now be described in more detail
in terms of features and operations with reference to the
accompanying drawings.
FIG. 1A
[0190] An embodiment of the present application is a
continuous-flow system for manufacturing pharmaceutical
formulations. An example of such a continuous-flow system is
presented in FIG. 1A. FIG. 1A is a schematic diagram of the
significant components and sub-systems used in a system for
manufacturing pharmaceutical formulations. Each of the components
and sub-systems can be included in, and operated as, a part of the
larger manufacturing system or may, in the alternative, be run
autonomously in a continuous fashion to accomplish the objective of
each sub-system as described herein. Additionally, each sub-system
may be run in a continuous-flow manner using batch inputs and
producing a batch output.
[0191] In one embodiment, the manufacturing system 100 is comprised
of a tank 5, which can hold a first fluid, the first fluid can be
pumped by a positive-displacement pump 2 through a hydrophilic
sterilizing filter 15 and into a high shear mixer 25. In some
embodiments, the first fluid can be an aqueous solution. In some
embodiments, the first fluid can be a first liquid. In some
embodiments, the first liquid can be an aqueous solution.
Similarly, tank 10 can feed a second fluid, via a
positive-displacement pump 12, through a hydrophobic sterilizing
filter 20 into the high shear mixer 25. In some embodiments, the
second fluid can be a second liquid. In some embodiments, the
second liquid can be an organic solvent. The high shear mixer 25, a
heat exchanger 30, and associated inlet line 96 and outlet line 97
comprise one embodiment of a first sub-system. In some embodiments,
the first sub-system can be a first emulsification sub-system. In
some embodiments, the high shear mixer 25 can create a first
dispersion of aqueous particles (the "discontinuous phase")
suspended in an organic continuous phase. In some embodiments, the
first dispersion can be a first component. In some embodiments, a
majority of the first component can then be fed from the high shear
mixer 25 into the heat exchanger 30, and the cooled first component
can flow back to the high shear mixer 25. In one embodiment,
however, a portion of the emulsion leaving the high shear mixer 25
can be forced towards a nozzle 75, for example by the addition of
additional organic solvent and aqueous solution to the first
emulsification sub-system.
[0192] In some embodiments, the emulsion leaving the high shear
mixer 25 can be transferred to an evaporation apparatus or
evaporation sub-system. In one embodiment the evaporation
apparatus, or evaporation sub-system, is comprised of a solvent
removal vessel 50, the atomizing nozzle 75, a gas inlet 115, and a
gas outlet 80. In some embodiments, the nozzle 75 acts to spray
atomized droplets of the first component into the solvent removal
vessel 50. In some embodiments, the nozzle 75 can combine the first
component with a fluid from a tank 60 and spray the first
component/liquid mixture into the solvent removal vessel 50 as
atomized droplets. In some embodiments, the fluid from the tank 60
is a liquid. In some embodiments, the liquid from the tank 60 is a
buffer solution. In some embodiments, the buffer solution can be
fed by a positive-displacement pump 22 from the tank 60 through a
hydrophilic sterile filter 65 into the nozzle 75. Gas from the gas
supply can be passed through a pressure regulator 11 and a
sterilizing gas filtration system 35 before entering the nozzle 75.
In some embodiments, the gas acts to atomize the first
component/liquid mixture as it exits the nozzle 75 into the solvent
removal vessel 50.
[0193] In some embodiments, a carrier gas can be supplied to the
solvent removal vessel 50 from the gas supply and enter the solvent
removal vessel through the gas inlet 115. The carrier gas can be,
for example, nitrogen gas (or a nitrogen gas/aqueous vapor mixture)
or air scrubbed of CO.sub.2. The pressure of the carrier gas can be
regulated using a pressure regulator 31. In some embodiments, the
carrier gas first passes through a heater/humidifier 90 and then
through a gas filtration system 45 before entering the solvent
removal vessel 50 through the gas inlet 115. In some embodiments,
the carrier gas circulates through the solvent removal vessel 50,
and creates an intense gas vortex, which can facilitate solvent
evaporation from the atomized droplets as they are sprayed into the
solvent removal vessel 50 from the nozzle 75. The gas vortex can
substantially prevent the atomized droplets from exiting the
sovlent removal chamber with the carrier gas through gas outlet 80.
With evaporation of the solvent, the atomized droplets become
droplets of large diameter synthetic membrane vesicles within an
aqueous phase. In some embodiments, the atomized droplets can be
first component/liquid mixture droplets. In some embodiments, the
large diameter synthetic membrane vesicles can be within an aqueous
phase. In some embodiments, the large diameter synthetic membrane
vesicles can be multivesicular liposomes. In some embodiments, the
carrier gas and evaporated solvent can then be removed from the
solvent removal vessel 50 through the gas outlet 80, after which
they can then pass through a filtration system 55 comprised of a
pre-filter and a sterilizing barrier filter, before being removed
as waste 95. In some embodiments, the prefilter (not shown) can be
a high efficiency cyclone separator.
[0194] In one embodiment, the evaporation sub-system is comprised
of a plurality of solvent removal vessels, each with one or more
atomizing nozzles, used in parallel to evaporate the solvent from
the atomized droplets. In some embodiments, the evaporation
sub-system can be comprised of a plurality of solvent removal
vessels, such as solvent removal vessel 50, each with one or more
atomizing nozzles, such as atomizing nozzle 75, used in parallel to
evaporate solvent from the atomized droplets. In some embodiments,
the solvent removal vessel can have additional multiple atomizing
nozzles (not shown) such as 75.
[0195] In some embodiments, a portion of the gas passing through
the heater/humidifier 90 can be directed towards a gas inlet 110
located in the lid of the solvent removal vessel 50. In some
embodiments, the gas inlet 110 allows gas to enter the solvent
removal vessel 50 and circulate in the upper portion of the vessel
acting to prevent particle buildup on the lid.
[0196] In some embodiments, a two-fluid rinse nozzle 105 can be
placed in and through the lid of the solvent removal vessel 50. In
some embodiments, the rinse nozzle 105 can spray the wall of
solvent removal vessel 50. The rinse nozzle 105, which receives a
buffer solution from a buffer solution tank 66, through a pump 64
and a sterilizing filter 62, can spray atomized tank wall rinse
solution particles into the solvent removal vessel 50. The buffer
solution can be atomized by gas traveling into the nozzle 105
through a pressure regulator 21 and a sterilizing gas filtration
system 85. Spraying atomized wall rinse solution particles into the
solvent removal vessel 50 can act to prevent the large diameter
synthetic membrane vesicles droplets (FIG. 7, component 7380) from
sticking to the walls of the solvent removal vessel 50 by rinsing
or flushing particles from the internal surfaces of the vessel 50,
and out a drain port 130 on the bottom of the vessel 50.
[0197] Removal of the solvent from the atomized droplets, affords
large diameter synthetic membrane vesicles coated in a buffer
solution shell as droplets. In some embodiments, the large diameter
synthetic membrane vesicles can be multivesicular liposomes. These
droplets then can collect on the bottom of the solvent removal
vessel 50 to form a suspension of large diameter synthetic membrane
vesicles in a buffer solution. This large diameter synthetic
membrane vesicles suspension can subsequently be pumped by a pump
125 through a solvent removal vessel outlet line 120 to an optional
sub-system 70. In some embodiments, the large diameter synthetic
membrane vesicles suspension can be pumped by a pump 125 through a
continuous heat treatment system 150 before entering the option
sub-system 70. In some embodiments, the buffer solution can be
exchanged for, for example, a saline solution. In some embodiments,
the sub-system 70 can be a single or series of concentration units
running in batch or continuous mode. In a typical embodiment, the
concentration units can be run in continuous mode. In a typical
embodiment, large diameter synthetic membrane vesicles can be
concentrated and the resulting suspension collected by the process.
In some embodiments, the large diameter synthetic membrane vesicles
can be multivesicular liposomes.
[0198] In some embodiments, one or more components of the
manufacturing system 100 can be omitted from the system.
FIG. 1B
[0199] In some embodiments, the manufacturing system 100 further
comprises a mass flow controller 13, a mass flow controller 23, a
mass flow controller 33, a rotometer flow indicator 57, a heater
93, a steam generator 40, and a metering pump 6 as shown in FIG.
1B. Each of the components and sub-systems can be included in, and
operated as, a part of the larger manufacturing system or may, in
the alternative, be run autonomously in a continuous fashion to
accomplish the objective of each sub-system as described herein.
Additionally, each sub-system may be run in a continuous-flow
manner using batch inputs and producing a batch output.
[0200] In some embodiments, the mass flow controllers 13, 23, and
33 measure, indicate, and control the gas flow supply to their
associated apparatus. In some embodiments, the rotometer flow
indicator 57 measures and indicates the gas flow to the lid
protection nozzle (FIG. 1A 110 7430. In some embodiments, the steam
generator 40 allows precise humidification of the carrier gas. In
some embodiments, the metering pump 6 allows precise control of the
water vapor generated by the steam generator 40.
FIG. 1C
[0201] One embodiment of the present application includes a
continuous heat treatment system. FIG. 1C provides a schematic of
one example of a continuous heat treatment system 150. In some
embodiments, the large diameter synthetic membrane vesicles
suspension can be pumped from the solvent removal vessel (FIG. 1A,
component 50) flowing through a feed line 120 (also seen in FIG.
1A, component 120), to the continuous heat treatment system 150
before entering a subsystem (FIG. 1A, component 70).
[0202] In some embodiments, the continuous heat treatment system
comprises a temperature controlled tank 151, optionally a
temperature control jacket (not shown), a pressurized tank 160, a
holding coil tubing 156, and a nitrogen source 157 as shown in FIG.
1C. Each of the components and sub-systems can be included in, and
operated as, a part of the larger manufacturing system or may, in
the alternative, be run autonomously in a continuous fashion to
accomplish the objective of each sub-system as described herein.
Additionally, each sub-system may be run in a continuous-flow
manner using batch inputs and producing a batch output.
[0203] In some embodiments, a portion of the large diameter
synthetic membrane vesicles suspension can be pumped by a pump
(FIG. 1A, component 125) through the feed line 120 into a mixing
vessel 180, for example an in-line static mixer. In some
embodiments, the large diameter synthetic membrane vesicles
suspension, flowing through the line 120, can come into contact
with a solution from the tank 151 before flowing through a line 155
into the mixing vessel 180. In one embodiment, the suspension
feeding the mixing vessel 180 through the line 120, flows at 165
ml/min and the solution through the line 155 flows at 247.5 ml/min.
In some embodiments, the solution can be fed through a line 152 by
a pump 153, the solution first passing through a hydrophilic
sterile filter 154 at a rate of 1.5 L per 1 L of the large diameter
synthetic membrane vesicles suspension added to the mixing vessel
180. In some embodiments, the tank 151 is temperature controlled.
In some embodiments, the solution from the tank 151 is a dextrose
solution. In some embodiments, the dextrose solution is heated to
about 98.degree. C. in the temperature controlled tank 151. In some
embodiments, the suspension/dextrose mixture flows through the
holding coil tubing 156. In some embodiments, the holding coil
tubing 156 holds the suspension/dextrose mixture for a specified
treatment time. In some embodiments, the treatment time is between
10 seconds and 30 seconds. In some embodiments, the
suspension/dextrose mixture is continuously heated for 30 seconds
to a temperature at or above 60C. In some embodiments, the holding
coil tubing 156 has a volume of 206 ml.
[0204] In some embodiments, the suspension/dextrose mixture can
leave the holding coil tubing 156 to enter a retentate vessel 168.
In some embodiments, a solution can be fed from the tank 160 into
the retentate vessel 168 through a line 164, the solution first
passing through a manual valve 162 and a sterilizing hydrophilic
filter 163, at a rate of 1.4 L per 1 L suspension/dextrose mixture
added to the retentate vessel 168. In some embodiments, the
solution flows through the line 164 at a rate of 578 ml/min. In
some embodiments, the tank 160 is temperature controlled. In some
embodiments, the temperature controlled tank is pressurized. In
some embodiments, the tank 160 is pressurized with the nitrogen
source 157 through a line 158 and a manual valve 159. In some
embodiments, the solution is a saline solution. In some
embodiments, the saline solution is a cold saline solution. In some
embodiments, the retentate vessel 168 is temperature controlled or
cooled.
[0205] In some embodiments, a portion of the suspension/dextrose
mixture in the retentate vessel 168 can be pumped by a pump 171
through a cross-flow (tangential-flow) filtration module 167. In
some embodiments, the permeate can be drawn off through a permeate
line 173 (passing through a sterilizing hydrophilic filter 172 and
a manual valve 174), wherein for each volume of suspension/dextrose
mixture added to the retentate vessel 168, a volume can be removed
and discarded. In some embodiments, the retentate from the
filtration module 167 can be circulated back into the retentate
vessel 168 via a retentate line 166. In some embodiments, for each
volume of suspension/dextrose mixture added to the retentate vessel
168, a volume of the liquid can be removed from the retentate
vessel 168 through a feed line 169 and a metering pump 170 to be
further processed by a subsystem (as seen in FIG. 1A, component 70;
systems of FIG. 8, FIG. 10 and FIG. 11). In some embodiments, the
suspension can flow through the pump 170 at a rate of 490
ml/min.
[0206] Filters 154, 163, and 172 are sterilizing hydrophilic
filters. Filters, 161 and 176 are sterilizing hydrophobic gas vent
filters used in the vessels and fed by gas lines 165 and 175,
respectively.
FIG. 2
[0207] One embodiment of the present application includes an
emulsification system, the process of using the system, and the
large diameter synthetic membrane vesicles products made by the
process. In some embodiments, the large diameter synthetic membrane
vesicles can be multivesicular liposomes. FIG. 2 provides a
schematic of one example of a continuous-flow emulsification system
210. The emulsification system 210 includes a high shear mixer 2130
and a heat exchanger 2170. In some embodiments, the high shear
mixer 2130 can be used in preparing a first component. The high
shear mixer 2130 used in one embodiment is the Ross HSM-703XS-20
Sanitary Inline High Shear Mixer made by the Charles Ross & Son
Company of Hauppauge, N.Y. In some embodiments, the head of the
high shear mixer 2130 can be connected to an aqueous inlet line
2120. In some embodiments, the head of the high shear mixer 2130
can be fed an aqueous solution through the aqueous inlet line 2120.
In some embodiments, the head of the high shear mixer 2130 can be
connected to a recirculation line 2125. In some embodiments, the
high shear mixer 2130 can feed a recirculated emulsion through the
recirculation line 2125. The recirculated emulsion contains
already-sheared aqueous droplets dispersed in a continuous organic
solution. In some embodiments, the aqueous solution can be stored
in a tank (FIG. 1A, component 10) and pass through a liquid
sterilization filter (FIG. 1A, component 20) attached to the
aqueous inlet line 2120 before entering the high shear mixer 2130
at various volumetric flow rates. High shear mixers are available
to deliver volumetric flow rates between 1 mL/minute and 4,000
mL/minute, between 10 mL/minute and 1,000 mL/minute, between 10
mL/minute and 100 mL/minute, and preferably between 15 mL/minute
and 50 mL/minute. In some embodiments, the aqueous inlet line 2120
can be configured to run co-axially through a portion of the
recirculation line 2125. The range of volumetric flow rates above
is appropriate when contemplating the use of one, or optionally
multiple, atomizing nozzles.
[0208] In some embodiments, the aqueous solution can be injected
through the center of a stator inside of the head of the high shear
mixer 2130 into the center of a spinning rotor (not shown) also
inside the high shear mixer head 2130. As the aqueous phase passes
between the rotor and the stator, inside the high shear mixer head
2130, it is sheared by the rotor teeth and stator teeth into
aqueous droplets. The sheared aqueous droplets become part of a
flowing stream of the recirculated emulsion which itself can be fed
back into the high shear mixer through the recirculation line 2125.
The combined aqueous solution and recirculated emulsion can pass
through blade shear gaps (not shown) in the high shear mixer head
2130, as it travels from the center to the outside of the rotor. In
one embodiment, on average, aqueous droplets are recirculated
approximately 100 times through the emulsification system 210, and
through that process, the aqueous droplets are sheared roughly
1,300 times by the blade shear gaps (not shown).
[0209] Exiting the high shear mixer head 2130 through an exit line
2150 is the resultant emulsion, with aqueous droplets dispersed in
the organic continuous phase, traveling at a volumetric flow rate
between 1 L/minute and 500 L/minute, between 5 L/minute and 100
L/minute, and preferably between 10 L/minute and 50 L/minute. The
major portion of the emulsion exiting the high shear mixer 2130,
approximately between 50% and 99.99%, between 70% and 99.9%, and
preferably between 80% and 99.9%, is passed through the heat
exchanger 2170 to cool the emulsion which has been heated by the
mechanical shearing process of the high shear mixer 2130.
Additionally, a smaller portion of the recirculating emulsion,
approximately between 1 mL/minute and 8,000 mL/minute, between 10
mL/minute and 2,000 mL/minute, between 20 mL/minute and 200
mL/minute, and preferably between 40 mL/minute and 100 mL/minute,
travels from the high shear mixer 2130 through a nozzle feed line
2180 to a nozzle for use in making the first component/buffer
atomized droplets. The range of volumetric flow rates above is
appropriate when contemplating the use of one, or optionally
multiple, atomizing nozzles. In some embodiments, the heat
exchanger 2170 can be comprised of a series of coils surrounded by
circulating solution. In some embodiments, the circulating solution
can be at a temperature ranging from about -5.degree. C. to about
30.degree. C. In some embodiments, the circulating solution can be
a glycol/water solution. In some embodiments, the glycol/water
solution can be at a temperature ranging from about 0.degree. C. to
about 10.degree. C. In some embodiments, the heat exchanger 2170 is
comprised of a series of coils surrounded by circulating 5.degree.
C. glycol/water solution. In some embodiments, the circulating
glycol/water solution can be fed to the heat exchanger 2170 through
an inlet line 2110 and an outlet line 2105. In some embodiments,
the emulsion travels through the heat exchanger coils, being cooled
by the glycol/water solution, and exits the heat exchanger through
a heat exchanger outlet 2175. In some embodiments, the heat
exchanger outlet 2175 can carry the cooled emulsion back through
the recirculation line 2125 for additional mixing with added
aqueous phase and added organic solution. In some embodiments, the
high shear mixer 2130 can act as a high volume centrifugal pump to
drive a high volumetric flow rate through the heat exchanger and
around the recirculation loop, back to the high shear mixer head
2130.
[0210] Attached to the recirculation line 2125 is an organic
solution inlet line 2160, which can be used to replenish the
organic solution in the emulsification system 210 according to the
volumetric amount of organic solution fed in the first component to
the nozzle feed line 2180. In some embodiments, the organic
solution can be stored in a tank (not shown) and flows, at a
volumetric flow rate between 1 mL/minute and 4,000 mL/minute,
between 10 mL/minute and 1,000 mL/minute, between 10 mL/minute and
100 mL/minute, and preferably between 20 mL/minute and 50
mL/minute. The range of volumetric flow rates above is appropriate
when contemplating the use of one, or optionally multiple,
atomizing nozzles. In some embodiments, the organic solution flows
through a liquid sterilization filter (shown in FIG. 1A and FIG.
1B, component 15) attached to the organic inlet line 2160 before
entering the recirculation line 2125 and the high shear mixer 2130.
In one embodiment, the volumetric flow rate of the organic solution
added through the inlet line 2160 can be equal to the volumetric
flow rate of aqueous solution added to the high shear mixer 2130
through the aqueous inlet line 2120. That is, in one embodiment,
the volumetric flow rates of the added liquids are at a 1:1 ratio.
At steady-state, because these are incompressible fluids and the
piping described herein is non-expandable piping, the flow rate of
the first component traveling to the nozzle through the nozzle feed
line 2180 is equal to the sum of the flow rates of the organic
solution being replenished to the emulsification system 210 through
the organic phase inlet line 2160 and the aqueous solution being
replenished to the emulsification system 210 through the aqueous
inlet line 2120.
[0211] In one embodiment, the emulsification system is used
autonomously to make MVL where the first component is collected in
a container and further processed to make the second emulsion. The
second emulsion is then sparged and filtered in a batch mode as
described in PCT Patent Publication No. WO99/13865 to S. Kim et
al., incorporated herein by reference.
[0212] In one embodiment, the emulsification system is employed
autonomously to produce common emulsions of triglycerides
(vegetable oils) and surfactants (Tween-20, Pluronic F-66, egg
lecithin) and water with common additives (scents, flavors). In an
additional embodiment, a cosmetic lotion is made with a
triglyceride as the discontinuous phase (oil in water emulsion). In
still other embodiments, the emulsification system produces
ointments, creams and salves, with the triglyceride as the
continuous phase. Additionally, a triglyceride emulsion in water
with egg yolk as the surfactant is used in the emulsification
system to produce a type of edible mayonnaise.
FIG. 3A-6
[0213] One embodiment of the present application includes an
atomizing nozzle, the process of using the nozzle, and the large
diameter synthetic membrane vesicles made by the process. In some
embodiments, the large diameter synthetic membrane vesicles can be
multivesicular liposomes. Examples of the instant atomizing
nozzles, the processes for using them, as well as the MVL products,
are presented in FIGS. 3A-6 and described herein.
FIG. 3A
[0214] FIG. 3A is a schematic partial view of an atomizing nozzle
310, providing a cross-sectional view of the lower portion of the
atomizing nozzle including a fluid contacting chamber 3125. The
fluid contacting chamber 3125 is conically tapered from the bottom
of an inner fluid conduit 3165 to a cylindrical tip 3145. The fluid
contacting chamber 3125 is annularly surrounded by an annular gas
chamber (a third fluid conduit) 3135, which narrows to form an
annular gas orifice 3150. The bottom portion of the cylindrical tip
3145 is annularly and concentrically surrounded by the annular gas
orifice 3150.
[0215] In some embodiments, a first fluid 3115 can travel at a
volumetric flow rate between 2 mL/minute and 1,000 mL/minute,
between 10 mL/minute and 500 mL/minute, between 20 mL/minute and
100 mL/minute, and preferably between 50 mL/minute and 100
mL/minute through an inner fluid conduit (central needle) 3165. In
a typical embodiment, the first fluid 3115 is an emulsion. In some
embodiments, the first fluid exits the bottom 3163 of the inner
fluid conduit 3165 and enters the fluid contacting chamber 3125,
whereupon the first fluid 3115 comes in physical communication with
a second fluid 3120. In a typical embodiment, the second fluid 3120
is a buffer solution. In some embodiments, the second fluid 3120
travels at a volumetric flow rate between 2 mL/minute and 1,000
mL/minute, between 10 mL/minute and 500 mL/minute, between 20
mL/minute and 100 mL/minute, and preferably between 50 mL/minute
and 100 mL/minute, through the outer fluid conduit 3123, which
annularly surrounds the inner fluid conduit 3165 (as seen from the
downward perspective view B and displayed in FIG. 3B), to reach the
fluid contacting chamber 3125. The above atomizing nozzle flow
rates are for a single atomizing nozzle. In some embodiments, the
first fluid 3115 forms a cylindrical core traveling through the
fluid contacting chamber 3125, now being in physical communication
with, and annularly surrounded by, the second fluid 3120. In some
embodiments, the first fluid and the second fluid are immiscible.
In some embodiments, the first fluid and second fluid can be
sparingly miscible. In some embodiments, the immiscibility between
the second fluid 3120 and the first fluid 3115, and the velocity at
which the second fluid 3120 and the first fluid 3115, causes the
second fluid 3120 to form a sheath around the first fluid 3115
core. In a typical embodiment, the first fluid 3115 is an emulsion
and the second fluid 3120 is a buffer solution.
[0216] In some embodiments, the diameter of the fluid contacting
chamber 3125 can be conically narrowed as it joins the cylindrical
tip 3145. As the first fluid 3115 travels through the narrowing
fluid contacting chamber 3125 and into the cylindrical tip 3145,
the diameter of the first fluid 3115 core is correspondingly
decreased. Likewise, the second fluid 3120 can be constricted to
create a thinner concentric annular sheath around the first fluid
3115 core. As a result of decreasing the diameter of the fluid
contacting chamber 3125 and passing the fluids through the
cylindrical tip 3145, the velocities of the first fluid 3115 and
second fluid 3120 are increased. In a typical embodiment, the first
fluid 3115 is an emulsion and the second fluid 3120 is a buffer
solution.
[0217] FIG. 3C shows an expanded cross-sectional view of the first
fluid and second fluid in the cylindrical tip 3145. In some
embodiments, the ratio of the diameters of the first fluid to the
second fluid in the nozzle tip can be the same as the ratio of the
diameters of the first fluid in inner fluid conduit 3165 to the
second fluid in the outer fluid conduit 3123. In some embodiments,
the ratio of the diameters of the first fluid to the second fluid
in the nozzle tip can be larger than the ratio of the diameters of
the first fluid in the inner fluid conduit 3165 to the second fluid
in the outer fluid conduit 3123. In some embodiments, the ratio of
the diameters of the first fluid to the second fluid in the nozzle
tip can be smaller than the ratio of the diameters of the first
fluid in inner fluid conduit 3165 to the second fluid in the outer
fluid conduit 3123.
[0218] FIG. 3D shows an expanded cross-sectional view of the first
fluid and second fluid in a droplet 3155. In some embodiments, the
ratio of the diameters of the first fluid to the second fluid in a
droplet 3155 can be the same as the ratio of the diameters of the
first fluid in the inner fluid conduit 3165 to the second fluid in
the outer fluid conduit 3123. In some embodiments, the ratio of the
diameters of the first fluid to the second fluid in a droplet 3155
can be larger than the ratio of the diameters of the first fluid in
the inner fluid conduit 3165 to the second fluid in the outer fluid
conduit 3123. In some embodiments, the ratio of the diameters of
the first fluid to the second fluid in a droplet 3155 can be
smaller than the ratio of the diameters of the first fluid in the
inner fluid conduit 3165 to the second fluid in the outer fluid
conduit 3123.
[0219] The nozzle 310 comprises a gas input channel 3130 which
supplies a gas 3140. In some embodiments, the volumetric flow rate
of the gas can be between 30 L/minute and 1,000 L/minute, between
30 L/minute and 500 L/minute, between 20 L/minute and 200 L/minute,
and preferably between 25 L/minute and 100 L/minute and at a
pressure between 5 psig and 1,000 psig, between 10 psig and 250
psig, between 20 psig and 150 psig, and preferably between 40 psig
and 120 psig to an annular gas chamber 3135. The above atomizing
nozzle flow rates are for a single atomizing nozzle. Prior to
reaching nozzle 310, the gas 3140 can be passed through a gas
filtration system (not shown) comprising a hydrophobic gas
sterilizer (as seen in FIG. 1A and FIG. 1b, component 35). The wall
of the fluid contacting chamber 3125 provides a barrier between the
annular gas chamber 3135 and the fluid contacting chamber 3125 such
that there is no physical contact between the fluids in the fluid
contacting chamber 3125 and the gas 3140 in the annular gas chamber
3135. In some embodiments, the outlet of the cylindrical tip 3145
and the outlet of the annular gas orifice 3150 can be flush with
each other such that second fluid 3120 does not come into contact
with the gas 3140 prior to the gas 3140 exiting the atomizing
nozzle 310. In other embodiments, the cylindrical tip 3145 extends
lower than the annular gas orifice 3150. In still other
embodiments, the cylindrical tip 3145 is slightly recessed within
the annular gas orifice 3150, such that the gas 3140 comes into
physical communication with the second fluid 3120 prior to the
second fluid fully exiting from the nozzle 310.
[0220] In some embodiments, as the gas 3140 passes through the
annular gas orifice 3150 and exits the atomizing nozzle 310, it
comes in physical communication with the second fluid 3120 and acts
to shear the stream of the second fluid 3120 and the first fluid
3115 into multiple atomized droplets 3155 made of a first fluid
3115 core and a second fluid 3120 shell. This configuration of the
three fluid nozzle can be best achieved when the cross sectional
areas of the inner fluid conduit 3165 and the second fluid path
within the fluid contacting chamber 3125 are chosen such that the
velocities of the first fluid 3115 and the second fluid 3120 are
approximately equal at the exit of the inner conduit 3163.
[0221] In some embodiments, the atomizing nozzle 310 can be
configured to produce very high yield large diameter synthetic
membrane vesicles (that is, vesicles that encapsulate a high amount
of a therapeutic agent). In some embodiments, the large diameter
synthetic membrane vesicles can be unilamilar vesicles or
multilamilar vesicles or polymer spheres encasing a liquid
comprising a therapeutic agent. In some embodiments, the large
diameter synthetic membrane vesicles can be multivesicular
liposomes. In some embodiments, the first fluid 3115 can be an
aqueous (or similar hydrophilic liquid) phase containing the
therapeutic agent and the second fluid 3120 can be an organic phase
comprising phospholipids or other encasing material e.g. a polymer
or PLGA (poly(lactic-co-glycolic acid), biocompatible and in-vivo
degradable polymer) or a wax. In some embodiments, the gas 3140
combines with an aqueous phase and an organic phase to afford a
droplet of aqueous phase inside a droplet of organic phase, which
can, for example, be used to form unilamilar vesicles or
multilamilar vesicles. In some embodiments, the size of the inner
aqueous phase droplet can have an average diameter of at least
about 0.5 .mu.m, 1 .mu.m, 5 .mu.m, 10 .mu.m, 15 .mu.m, 20 .mu.m, or
50 .mu.m, or a diameter within a range defined by any of two of the
preceding values. In some embodiments, the organic solvent can be
removed by using a normal spray drying system. In some embodiments,
the organic solvent can be removed by using solvent removal chamber
such as the one described in FIG. 7.
[0222] In some embodiments, the nozzle 310 can comprise an
additional outer fluid conduit (not shown), which annularly
surrounds the inner conduit 3165 to provide a four fluid atomizing
nozzle as shown in FIGS. 3E-3L. In some embodiments, the conduit
which annularly surrounds the inner conduit 3165 can include an an
aqueous phase suitable as a suspending medium. In some embodiments,
the nozzle can be operated to produce an emulsion where an aqueous
drug core can be surrounded by phospholipids in a volatile solvent
that in turn is surrounded by another aqueous phase, which can, for
example, be used to form unilamilar vesicles or multilamilar
vesicles. In some embodiments, the large diameter synthetic
membrane vesicles will initially have a hydrophobic outer
surface.
[0223] In some embodiments, the nozzle 310 can omit the conduit
3123 in order to provide a two fluid nozzle (not shown). In some
embodiments, the two fluids are a liquid and a gas. In some
embodiments, the two fluid nozzle (not shown) can be used to spray
a first component into the instant solvent removal chamber such as
the one described in FIG. 7. In some embodiments, the organic
solvent can be removed by contacting the first component with an
inert gas to in order to create the MVL. In some embodiments excess
phospholipids can be used in the first component to create the MVL
having a coating of phospholipids which initially has a hydrophic
surface and can partially shed to afford the MVL with a hydrophilic
surface.
FIG. 3E through FIG. 3L
[0224] Liposomes with high incorporation percentages (especially
important for an expensive active agents) can be made with a four
fluid atomizing nozzle 320 as depicted in FIG. 3E through FIG. 3L.
In some embodiments, a first fluid 3115 can be a fluid comprising
an aqueous active agent, while a second fluid 3170 can be a
volatile solvent comprising a lipid solution, wherein the second
fluid 3170 forms a membrane while a third fluid 3120 can be a
suspending buffer. Without the suspending buffer outer layer, the
lipids would for with their hydrophobic regions facing out towards
the hydrophobic gas. This four fluid nozzle assembles the outer
membrane in a way to prevent aggregation. In such embodiments, the
incorporation of the active agent is high because the active agent
fluid can be surrounded by the lipid layer forming solvent 3170.
Small concentrations of lipids in 3170 can provide at least one
bilayer. In some embodiments, higher concentrations of lipid in the
second fluid can provide liposomes comprising thick large numbers
of bilayers. In some embodiments, structurally strong and stable
liposomes can be made using tight packing saturated phospholipids
with a high transition temperature as they are deposited from an
evaporating solvent solution and do not have to be formed in
aqueous solutions as per conventional liposome processes.
[0225] When the first core fluid 3115 is the first component
lacking any lipids that could interact with the active agent (e.g.
charged lipid like DPPG) and 3170 is volatile solvent (could be
same or different that used in first component) and 3120 is a
suspending buffer; MVLs with different lipids on the outside. These
outside lipids (from 3170) can be highly charged for stability or
long chain saturated for mechanical strength. The lipids can be put
on the MVL as 1 or 2 bilayers or, if the lipid concentration in
3170 is high, be put on as a thick mechanically strong many
multiple-bilayers (not previously done) to increase stability of
the MVL and provide longer slower active agent release. If polymers
(e.g. PLGA) were also added to 3120 (w/wo lipids), a polymer
skeleton could be formed surrounding the MVL, further stabilizing
it physically and chemically. If the polymer layer were strong
enough. it would allow the MVLs to be lyophilized and be stable for
very long times at room temperature.
[0226] FIG. 3E is a schematic partial cross-sectional view of a
four fluid atomizing nozzle 320, comprising a first inner fluid
conduit (central needle) 3165, a second inner conduit 3175, an
outer conduit 3123, a gas input channel 3130, an annular gas
chamber (a fourth fluid conduit) 3135, a first fluid contacting
chamber 3125, a second fluid contacting chamber 3137, and an
annular gas orifice 3150. The first fluid contacting chamber 3137
is conically tapered from the bottom of the first inner fluid
conduit 3165 and the exit orfice 3147 of the second inner fluid
conduit 3175. The second fluid contacting chamber 3137 is conically
tapered from the exit orfice 3147 of the second inner fluid conduit
3175 to a cylindrical tip 3145. In some embodiments, the exit
orfice 3147 of the second inner fluid conduit 3175 can be between
the exit orfice of the first inner fluid conduit 3163 and the
cylindrical tip 3145.
[0227] In some embodiments, the second fluid contacting chamber,
allows for the first and second fluids to contact the third fluid
3120 for a shorter duration than the nozzle described in FIG. 3I.
In some embodiments, the four fluid atomizing nozzle 320 depicted
in FIG. 3E can handle flow rates higher than the one fluid
contacting chamber nozzle described in FIG. 3I through FIG. 3L due
to lower flow instabilities. In some embodiments, the first and
second fluid contacting chambers are hydraulic extrusion cones.
[0228] The second fluid contacting chamber 3137 is annularly
surrounded by the annular gas chamber 3135, which narrows to form
an annular gas orifice 3150. The exit orfice portion of the
cylindrical tip 3145 is annularly and concentrically surrounded by
the annular gas orifice 3150.
[0229] In some embodiments, a first fluid 3115 can travel at a
volumetric flow rate between 2 mL/minute and 1,000 mL/minute,
between 10 mL/minute and 500 mL/minute, between 10 mL/minute and
100 mL/minute, and preferably between 50 mL/minute and 100
mL/minute through the first inner fluid conduit 3165. In some
embodiments, the first fluid exits the exit orfice 3163 of the
first inner fluid conduit 3165 and enters the first fluid
contacting chamber 3125, whereupon the first fluid 3115 comes in
physical communication with the second fluid 3170. In some
embodiments, the second fluid 3170 travels at a volumetric flow
rate between 2 mL/minute and 1,000 mL/minute, between 10 mL/minute
and 500 mL/minute, between 10 mL/minute and 100 mL/minute, and
preferably between 50 mL/minute and 100 mL/minute, through a second
inner fluid conduit 3175. In a typical embodiment, the first fluid
3115 is an emulsion without any lipids that could interact with the
active agent (e.g. no charged lipid such as DPPG) In another
embodiment, the second fluid 3170 is a volatile solvent comprising
a lipid solution (could be the same or different than that used in
the first component).
[0230] In some embodiments, the first fluid 3115 travels through
the first fluid contacting chamber 3125, surrounded by the second
fluid 3170. In some embodiments, the first fluid 3115 and the
second fluid 3170 continue through the first fluid contacting
chamber 3125 into the second fluid contacting chamber 3137. In some
embodiments, the second fluid 3170 forms a sheath around the first
fluid 3115 core.
[0231] In some embodiments, the first and second fluids can travel
at a volumetric flow rate between 2 mL/minute and 1,000 mL/minute,
between 10 mL/minute and 500 mL/minute, between 10 mL/minute and
100 mL/minute, and preferably between 50 mL/minute and 100
mL/minute through the second inner conduit 3175. In some
embodiments, the first and second fluids exits the exit orfice 3147
the second inner conduit 3175 and enter the second fluid contacting
chamber 3137, whereupon the first and second fluids come in
physical communication with a third fluid 3120. In a typical
embodiment, the third fluid 3120 is a buffer solution. In some
embodiments, the third fluid 3120 travels at a volumetric flow rate
between 2 mL/minute and 1,000 mL/minute, between 10 mL/minute and
500 mL/minute, between 10 mL/minute and 100 mL/minute, and
preferably between 50 mL/minute and 100 mL/minute, through an outer
fluid conduit 3123, which annularly surrounds the first inner fluid
conduit 3165 and the second inner conduit 3175 (as seen from the
downward perspective view F and displayed in FIG. 3F). In some
embodiments, the first, second and third fluids exit the exit
orfice 3145 of the second fluid contacting chamber 3137. In some
embodiments, the third fluid 3120 forms a sheath around the first
and second fluids.
[0232] In some embodiments, the diameter of the first fluid
contacting chamber 3125 can be conically narrowed as it approaches
the exit orfice 3163 of the first inner fluid conduit 3165. As the
first fluid 3115 travels through the narrowing first fluid
contacting chamber 3125 and toward the exit orfice 3147 of the
second fluid conduit 3175, the diameter of the first fluid 3115
core is correspondingly decreased Likewise, the second fluid 3170
can be constricted to create a thinner concentric annular sheath
around the first fluid 3115 core. As a result of decreasing the
diameter of the first fluid contacting chamber 3125 and passing the
fluids through the exit orfice 3147 of the second fluid conduit
3175, the velocities of the first fluid 3115 and the second fluid
3170 are increased.
[0233] In some embodiments, the diameter of the second fluid
contacting chamber 3125 can be conically narrowed as it joins the
cylindrical tip 3145. As the first fluid 3115 core and second fluid
3120 travel through the narrowing second fluid contacting chamber
3125 into the cylindrical tip 3145, the diameter of the first fluid
3115 core, the second fluid 3120 the third fluid 3170 can be
further correspondingly decreased. Likewise, the third fluid 3170
can be constricted to create a thinner concentric annular sheath
around the first fluid 3115 core and the second fluid 3120 sheath.
In some embodiments, the multi vesicular liposomes are lipid coated
with lipids on the outside that are different than the lipids on
the inside. In some embodiments, the four fluid nozzle illustrated
in FIG. 3E and FIG. 3I could be used to manufacture multi vesicular
liposomes coated with an additional layer of polymer PLGA or a
highly charged phospholipid different from the phospholipid of the
internal membrane. These multi vesicular liposomes coated with an
extra layer of polymer or phospholipid or combination thereof
constitute further embodiments as disclosed herein.
[0234] The four fluid atomizing nozzle 320 comprises a gas input
channel 3130 which supplies a gas 3140. In some embodiments, the
volumetric flow rate of the gas can be between 30 L/minute and
1,000 L/minute, between 30 L/minute and 500 L/minute, between 20
L/minute and 200 L/minute, and preferably between 25 L/minute and
100 L/minute and at a pressure between 5 psig and 1,000 psig,
between 10 psig and 250 psig, between 20 psig and 150 psig, and
preferably between 50 psig and 120 psig to the annular gas chamber
3135. The above atomizing nozzle flow rates are for a single
atomizing nozzle. Prior to reaching the nozzle 320, the gas 3140
can be passed through a gas filtration system (not shown)
comprising a hydrophobic gas sterilizer (as seen in FIG. 1A and
FIG. 1B, component 35). In some embodiments, the outlet of the
cylindrical tip 3145 and the outlet of the exit orfice 3147 of the
first second contacting chamber 3137 have the same diameter. In
some embodiments, the diameter of the outlet of the cylindrical tip
3145 is larger than the diameter of the outlet of the exit orfice
3147 of the second fluid contacting chamber 3137. In some
embodiments, the diameter of the outlet of the cylindrical tip 3145
is smaller than the diameter of the exit orfice 3147 of the second
fluid contacting chamber 3137.
[0235] FIG. 3G shows an expanded cross-sectional view of the first
fluid, second fluid and third fluid in the cylindrical tip
3145.
[0236] FIG. 3H shows an expanded cross-sectional view of the first
fluid, second fluid and third fluid in a droplet 3255
FIG. 3I
[0237] FIG. 3I is a schematic partial cross-sectional view of a
four fluid atomizing nozzle 320, comprising a first inner fluid
conduit 3165, a second inner conduit 3175, an outer conduit 3123, a
fluid contacting chamber 3125, a gas input channel 3130, an annular
gas chamber (a fourth fluid conduit) 3135, and an annular gas
orifice 3150. In some embodiments, the fluid contacting chamber
3125 is conically tapered from the exit orfice 3173 of an outer
fluid conduit 3123 to a cylindrical tip 3145.
[0238] The fluid contacting chamber 3125 is annularly surrounded by
an annular gas chamber 3135, which narrows to form an annular gas
orifice 3150. The bottom portion of the cylindrical tip 3145 is
annularly and concentrically surrounded by the annular gas orifice
3150.
[0239] In some embodiments, a first fluid 3115 can travel at a
volumetric flow rate between 2 mL/minute and 1,000 mL/minute,
between 10 mL/minute and 500 mL/minute, between 10 mL/minute and
100 mL/minute, and preferably between 50 mL/minute and 100
mL/minute through the first inner fluid conduit 3165. In a typical
embodiment, the first fluid 3115 is an emulsion. In some
embodiments, the first fluid exits the bottom 3163 of the first
inner fluid conduit 3165 and enters the fluid contacting chamber
3125, whereupon the first fluid 3115 comes in physical
communication with a second fluid 3170. In some embodiments, a
second fluid 3170 exits the bottom 3173 of the second inner conduit
3175 and enters the fluid contacting chamber 3125, whereupon the
second fluid 3170 comes in physical communication with the first
fluid 3115 and a third fluid 3170. In a typical embodiment, the
first fluid 3115 in the first inner fluid conduit 3165 is annularly
surrounded by the second fluid 3170 in the second inner conduit
3175 which is annularly surrounded by the third fluid 3120 in the
outer conduit 3123 (as seen from the downward perspective view J
and displayed in FIG. 3J).
[0240] In some embodiments, the first fluid 3115 travels through
the first fluid contacting chamber 3125, surrounded by the second
fluid 3170 which is surrounded by the third fluid 3120. In some
embodiments, the second fluid 3170 forms a sheath around the first
fluid 3115 and the third fluid 3120 forms a sheath around the first
and second fluids.
[0241] In some embodiments, the diameter of the fluid contacting
chamber 3125 can be conically narrowed as it joins the cylindrical
tip 3145. In some embodiments, as the first fluid 3115 travels
through the narrowing first fluid contacting chamber 3125 and
toward the cylindrical tip 3145, the diameter of the first fluid
3115 core is correspondingly decreased. In some embodiments, the
second fluid 3170 can be constricted to create a thinner concentric
annular sheath around the first fluid 3115 core. In some
embodiments, the third fluid 3120 can be constricted to create a
thinner concentric annular sheath around the first fluid 3115 core
and the concentric annular second fluid 3170. As a result of
decreasing the diameter of the first fluid contacting chamber 3125
and passing the fluids through the cylindrical tip 3145, the
velocities of the first fluid 3115, second fluid 3170 and third
fluid 3120 can be increased.
[0242] The four fluid atomizing nozzle 320 comprises a gas input
channel 3130 which supplies a gas 3140. In some embodiments, the
volumetric flow rate of the gas can be between 30 L/minute and
1,000 L/minute, between 30 L/minute and 500 L/minute, between 20
L/minute and 200 L/minute, and preferably between 25 L/minute and
100 L/minute and at a pressure between 5 psig and 1,000 psig,
between 10 psig and 250 psig, between 20 psig and 150 psig, and
preferably between 40 psig and 120 psig to the annular gas chamber
3135. The above atomizing nozzle flow rates are for a single
atomizing nozzle. Prior to reaching the nozzle 320, the gas 3140
can be passed through a gas filtration system (not shown)
comprising a hydrophobic gas sterilizer (as seen in FIG. 1A and
FIG. 1B, component 35).
[0243] FIG. 3K shows an expanded cross-sectional view of the first
fluid, second fluid and third fluid in the cylindrical tip
3145.
[0244] FIG. 3L shows an expanded cross-sectional view of the first
fluid, second fluid and third fluid in a droplet 3255.
FIG. 4A
[0245] FIG. 4A depicts an atomizing nozzle 410 where the first
fluid 4115 can break down into large droplets in the fluid
contacting chamber 4125 prior to reaching the cylindrical tip
4145.
[0246] In some embodiments, a first fluid 4115 exits a inner fluid
conduit 4165 through the exit orfice 4163 of the inner fluid
conduit 4165 and enters the fluid contacting chamber 4125,
whereupon the first fluid 4115 comes in physical communication with
a second fluid 4120. The second fluid 4120 travels through an outer
fluid conduit 4123, which annularly surrounds the inner fluid
conduit 4165 (as seen from the downward perspective view B and
displayed in FIG. 4B), to reach the fluid contacting chamber 4125.
The first fluid 4115 forms a plurality of first fluid droplets 4157
traveling through the fluid contacting chamber 4125, the first
fluid droplets 4157 now being in physical communication with, and
surrounded by, the second fluid 4120. In some embodiments, the
first and second fluid are immiscible. In a typical embodiment, the
first fluid 4115 is an emulsion and the second fluid 4120 is a
buffer solution.
[0247] In some embodiments, the diameter of the fluid contacting
chamber 4125 is conically narrowed as it joins the cylindrical tip
4145. As the first fluid 4115 travels through the narrowing fluid
contacting chamber 4125 and into the cylindrical tip 4145, the
first fluid droplets 4157 are squeezed and their diameter is
correspondingly decreased along the axis of travel. Likewise, the
second fluid 4120 is constricted to create a thinner shell around
the first fluid droplets 4157. As a result of decreasing the
diameter of the fluid contacting chamber 4125 and passing the
solutions through the cylindrical tip 4145, the velocities of the
first fluid droplets 4157 and second fluid 4120 are increased. In a
typical embodiment, the first fluid 4115 is an emulsion and the
second fluid 4120 is a buffer solution.
[0248] FIG. 4C shows an expanded cross-sectional view of the first
fluid droplets 4157 and second fluid 4120 in the cylindrical tip
4145.
[0249] FIG. 4D shows an expanded cross-sectional view of the first
fluid and second fluid in a droplet 4155. In some embodiments, the
ratio of the diameters of the first fluid to the second fluid in a
droplet 4155 can be the same as the ratio of the diameters of the
first fluid in the inner fluid conduit 4165 to the second fluid in
the outer fluid conduit 4123. In some embodiments, the ratio of the
diameters of the first fluid to the second fluid in a droplet 4155
can be the larger than the ratio of the diameters of the first
fluid in the inner fluid conduit 4165 to the second fluid in the
outer fluid conduit 4123. In some embodiments, the ratio of the
diameters of the first fluid to the second fluid in a droplet 4155
can be the smaller than the ratio of the diameters of the first
fluid in the inner fluid conduit 4165 to the second fluid in the
outer fluid conduit 4123.
[0250] The nozzle 410 comprises a gas input channel 4130 which
supplies a gas 4140 to an annular gas chamber (a third fluid
conduit) 4135. Prior to reaching the nozzle 410, the gas 4140 can
be passed through a gas filtration system (not shown) comprising a
hydrophobic gas sterilizer (FIG. 1A and FIG. 1B, component 35). The
wall of the fluid contacting chamber 4125 provides a barrier
between the annular gas chamber 4135 and the fluid contacting
chamber 4125 such that there is no physical contact between the
fluids in the fluid contacting chamber 4125 and the gas 4140 in the
annular gas chamber 4135. In some embodiments, the outlet of the
cylindrical tip 4145 and the outlet of the annular gas orifice 4150
can be flush with each other such that the second fluid 4120 does
not come into contact with the gas 4140 prior to the gas 4140
exiting the atomizing nozzle 410. In other embodiments, the
cylindrical tip 4145 may extend below the end of the annular gas
orifice 4150. In still other embodiments, the cylindrical tip 4145
may be slightly recessed within the annular gas orifice 4150, such
that the gas 4140 comes into physical communication with the second
fluid 4120 prior to the second fluid fully exiting from the nozzle
410. In a typical embodiment, the first fluid 4115 is an emulsion
and the second fluid 4120 is a buffer solution.
[0251] In some embodiments, as the gas 4140 passes through the
annular gas orifice 4150 and exits the atomizing nozzle 410, it
comes in physical communication with the second fluid 4120 and acts
to shear the stream of the second fluid 4120 and the first fluid
4115 droplets into atomized droplets 4155 made of a first fluid
4115 core and a second fluid 4120 shell. In this embodiment the
first fluid core stream breaks up due to the mismatch of velocities
between the first fluid 4115 and the second fluid 4120 at the exit
orfice 4163 of the inner fluid conduit 4165. In a typical
embodiment, the first fluid 4115 is an emulsion and the second
fluid 4120 is a buffer solution.
FIG. 5
[0252] FIG. 5 is a more detailed view of one embodiment of an
atomizing nozzle 505. The nozzle depicted in FIG. 5 is derived from
Part No. 1/8JJN-SS+SUJ1A-SS manufactured by Spraying Systems Co. of
Wheaton, Ill., as modified in FIG. 5 and described herein. The
atomizing nozzle lower section 510 of FIG. 5 provides three fluid
input channels for the introduction of three fluids into the nozzle
505. A first fluid input channel is the central longitudinal
channel 5114, which extends substantially downwardly through the
atomizing nozzle lower section 510 and extends into a fluid
contacting chamber 5125. In some embodiments, a second fluid input
channel is a buffer input channel 5128 and a third fluid input
channel is a gas input channel 5130.
[0253] The central longitudinal channel 5114 is a channel which may
be of circular cross-section. The central longitudinal channel 5114
may begin as an aperture 5111 in a line coupler 5113. The line
coupler 5113 is a means for attaching a supply line to the nozzle
505 for the introduction of the first fluid 5115 into the nozzle
lower section 510. A polymeric sealing washer 5112 is retained in
contact with the line coupler 5113 and interposed between a coupler
nut 5116 of the line coupler 5113 and a head (not shown) of a inner
fluid conduit 5165.
[0254] The inner fluid conduit 5165 can provide a continuation of
the central longitudinal channel 5114, with a narrowed central bore
5164 extending through a removable housing member 5170 into the
horizontal channel casing 5175, and into the liquid fluid cap 5180,
ending in the fluid contacting chamber 5125 of the liquid fluid cap
5180. In some embodiments, the housing member 5170 can be threaded.
The housing member 5170 can be fitted on the top with a housing cap
5172 and housing gasket member 5171 to provide secure fastening and
fluid seal. The housing member 5170 extends downwardly into a top
portion of the horizontal channel casing 5175, wherein it can be
tightened to form a secure connection with the horizontal channel
casing 5175, for example using internal screw threading.
[0255] In addition to the central longitudinal channel 5114, the
horizontal channel casing 5175 can be comprised of the gas input
channel 5130 and the buffer input channel 5128. The gas input
channel 5130 can have an external aperture 5131 to which a gas
supply line (not shown) can be to be attached. The gas input
channel 5130 can have an internal aperture 5132 which leads to an
annular gas chamber (third fluid conduit) 5135, the annular gas
chamber 5135 situated to annularly surround the liquid fluid cap
5180. The liquid fluid cap 5180 can be detachably and securely
fitted into the horizontal channel casing 5175 such that fluid
contents of the liquid fluid cap 5180 and the fluid contacting
chamber 5125 cannot physically communicate with the fluid contents
of the annular gas chamber 5135 inside the nozzle. The buffer input
channel 5128 can have an external aperture 5127 to which a fluid
supply line (not shown) can be attached. The buffer input channel
5128 can have an internal aperture 5126 which leads to the top
opening of the liquid fluid cap 5180.
[0256] In some embodiments, the liquid fluid cap 5180 can be
cylindrically shaped in a top portion and conically tapered in a
exit orfice portion. In some embodiments, the liquid fluid cap 5180
can be cylindrically tapered in both a top portion and a bottom
portion. In some embodiments, the fluid contacting chamber 5125 and
the outer fluid conduit 5123 form the interior wall of the liquid
fluid cap 5180. The fluid contacting chamber 5125 is conically
tapered from the exit orfice of an annular outer fluid conduit 5123
to a cylindrical tip 5145. The exit orfice portion of the
cylindrical tip 5145 can be annularly and concentrically surrounded
by an annular gas orifice 5150. The annular gas orifice 5150 can be
created by placement of the cylindrical tip 5145 in the center of a
cylindrical opening in a gas cap 5160. The liquid fluid cap 5180
can have a gas restrictor 5182 attached to the exterior of the
liquid fluid cap 5180 with several circular holes in the gas
restrictor 5182, which annularly surrounding the liquid fluid cap
5180. The presence of the gas restrictor 5182 creates a lower
annular gas chamber 5136 between the gas restrictor 5182 and the
gas cap 5160. The gas restrictor 5182 can minimize the turbulence
possibly created as a gas travels through the annular gas chamber
5135 to the annular gas orifice 5150. The nozzle lower section 510
can be fitted to an orifice housing 5185 which acts to keep the gas
cap 5160 and liquid fluid cap 5180 securely fitted to the
horizontal channel casing 5175. In some embodiments, the orifice
housing 5185 can be threaded.
[0257] In some embodiments, the first fluid 5115 can enter the
longitudinal channel 5114 of the nozzle lower section 510 from a
supply line (not shown) attached to the line coupler 5113. The
first fluid travels through the longitudinal channel 5114 until it
reaches the fluid contacting chamber 5125 whereupon it comes in
physical contact with the second fluid 5120 in the fluid contacting
chamber 5125. In a typical embodiment, the first fluid 5115 is an
emulsion and the second fluid 5120 is a buffer solution.
[0258] In some embodiments, the second fluid 5120 enters the nozzle
lower section 510 through an input aperture 5127 and travels
through the input channel 5128. In some embodiments, the second
fluid 5120 then passes through the internal aperture 5126 leading
to the top of the liquid fluid cap 5180 and travels through the
annular outer fluid conduit 5123, and travels around and surrounds
the inner fluid conduit 5165 at a lower section to reach the fluid
contacting chamber 5125. In some embodiments, upon exiting the exit
orfice 5163 of the inner fluid conduit 5165 or the central
longitudinal channel 5114 and entering the fluid contacting chamber
5125, the first fluid 5115 can form a cylindrical core traveling
through the fluid contacting chamber 5125. In some embodiments, the
first fluid 5115 cylindrical core can be surrounded annularly by
the second fluid 5120. In some embodiments, the immiscibility
between the second fluid 5120 and the first fluid 5115, and because
of the high velocity at which the second fluid 5120 and first fluid
5115 are traveling, causes the second fluid 5120 to form a sheath
around the first fluid 5115 core. The first fluid 5115 and second
fluid 5120 can travel through the fluid contacting chamber 5125
into the cylindrical tip 5145 of the outer conduit 5123. In a
typical embodiment, the first fluid 5115 is an emulsion and the
second fluid 5120 is a buffer solution.
[0259] The cross-sectional diameter of the fluid contacting chamber
5125 decreases as it conically narrows to join the cylindrical tip
5145. As the first fluid 5115 travels through the narrowing fluid
contacting chamber 5125 and into the cylindrical tip 5145 the
diameter of the first fluid 5115 core is decreased. Likewise, the
second fluid 5120 can be constricted to create a thinner concentric
annular sheath around the first component 5115 core. In some
embodiments, as the solutions pass through the narrowing
cylindrical tip, the velocities of the first fluid 5115 and the
second fluid 5120 can be increased. In some embodiments, the
threads on the inner fluid conduit 5165 and the housing member 5170
allow the outlet tip of the inner fluid conduit 5165 to be adjusted
vertically within the conical portion of the fluid contacting
chamber 5125 within the liquid fluid cap 5180. In some embodiments,
the housing member 5170 can be adjusted so that the velocities of
the second fluid 5120 and the first fluid 5115 can be made
approximately equal at the point where the first fluid 5115 exits
the inner fluid conduit 5165. This allows optimal operation as
shown in FIG. 3A. In some embodiments, the housing member 5170 can
be adjusted so that the velocities of the second fluid 5120 and the
first fluid 5115 can be made uequal at the point where the first
fluid 5115 exits the inner fluid conduit 5165. This allows optimal
operation as shown in FIG. 4A. In a typical embodiment, the first
fluid 5115 is an emulsion and the second fluid 5120 is a buffer
solution.
[0260] In some embodiments, a gas 5140 can be first passed through
a gas filtration system (not shown) comprising a hydrophobic gas
sterilizer (not shown). In some embodiments, the gas 5140 can enter
the gas input channel 5130 through the external gas input aperture
5131 and then passes through the internal gas chamber aperture 5132
to reach the annular gas chamber 5135. In some embodiments, the
liquid fluid cap 5180 can be detachably and securely fitted into
the horizontal channel casing 5175 such that the gas 5140 cannot
physically communicate with the first fluid 5115 or the second
fluid 5120 inside the nozzle lower section 510. The gas 5140
travels through the holes in the gas restrictor 5182 to reach the
lower annular gas chamber 5136.
[0261] As the gas 5140 passes through the annular gas orifice 5150
and exits the atomizing nozzle lower section 510, it comes in
physical communication with the second fluid 5120 and acts to shear
the stream of the second fluid 5120 and first fluid 5115 into
atomized droplets 5155 made of a first fluid 5115 core and a second
fluid 5120 shell. In a typical embodiment, the first fluid 5115 is
an emulsion and the second fluid 5120 is a buffer solution. In a
typical embodiment, the first fluid 5115 core is an emulsion and
the second fluid 5120 shell is a buffer solution.
[0262] Some embodiments provide an atomizing nozzle apparatus,
comprising three channels, each having at least one entrance
orifice and one exit orifice, the channels comprise of an inner
fluid conduit and an outer fluid conduit, wherein the exit orifice
for the inner fluid conduit is of a diameter smaller than, and is
located centrally in, the outer fluid conduit and is directed
towards the outer fluid conduit exit orifice, a gas channel,
wherein a pressurized gas exiting the gas channel exit orifice
impinges a liquid exiting the exit orifice of the outer fluid
conduit. Additional embodiments include the processes for using
such a device and the MVL products made by the same. In some
embodiments, the gas channel exit orifice can annularly surround
the exit orifice of the outer fluid conduit. In some embodiments,
the gas channel exit orifice and the outer fluid conduit exit
orifice can be flush. In some embodiments, the outer fluid conduit
exit orifice can be recessed within the gas channel exit orifice.
In some embodiments, the outer fluid conduit exit orifice can
extend beyond the gas channel exit orifice. In some embodiments,
the exit orifices of all three channels can be coaxial. In some
embodiments, the exit orifice of the inner fluid conduit can be
more than two outer fluid conduit exit orifice diameters away from
the outer fluid conduit exit orifice.
[0263] In some embodiments, the multi vesicular liposome product
produced by the atomizing nozzle in FIGS. 3-6 can be directly
collected and purified. Alternatively, the MULTI VESICULAR LIPOSOME
product can be processed by an evaporation tower or the evaporation
apparatus of FIG. 7 (or as part of FIGS. 1A-1C) before being
processed.
FIG. 6
[0264] FIG. 6 is an exploded schematic 605 of the individual
components of the device of FIG. 5.
[0265] In some embodiments, component 5113 is a line coupler;
component 5165 is an inner fluid conduit; component 5172 is a
housing cap; component 5170 is a housing member; component 5171 is
a housing gasket member; component 5180 is a liquid fluid cap;
component 5145 is a cylindrical tip; component 5160 is a gas cap;
component 5182 is a gas restrictor; and component 5185 is an
orifice housing.
[0266] In some embodiments, location 5130 can be a gas input
channel; location 5132 can be an internal gas chamber aperture;
location 5128 can be a fluid channel; location 5126 can be an
internal fluid cap aperture; and location 5150 can be an annular
gas orifice.
FIG. 7
[0267] An embodiment of the present application is a system for
manufacturing formulations including an evaporation apparatus, or
evaporation sub-system, the process of using the system, and the
large diameter synthetic membrane vesicles products made by the
process. An example of an evaporation apparatus is the solvent
removal vessel presented in FIG. 7 and described herein. FIG. 7
shows a schematic of a solvent removal vessel 710. In some
embodiments, atomized droplets 7155 can be sprayed into the solvent
removal vessel 710 in order to remove solvent from atomized
droplets 7155. In some embodiments, the solvent removal vessel 710
can include a three-fluid atomizing nozzle 7510, described above,
attached to and extending through a lid 7220 of the solvent removal
vessel 710. In some embodiments, the cylindrical tip and annular
gas orifice of the atomizing nozzle 7510 can be configured to be
substantially flush with the inside edge of the lid 7220. In some
embodiments, the cylindrical tip and annular gas orifice of the
atomizing nozzle 7510 can be configured to extend beyond the inside
edge of the lid 7220.
[0268] Alternatively, in some embodiments, the solvent removal
vessel 710 can include a three-fluid atomizing nozzle 7510,
described above, attached to and extending through a wall 7350 of
the solvent removal vessel 710. In some embodiments, the
cylindrical tip and annular gas orifice of the atomizing nozzle
7510 can be configured to be substantially flush with the inside
edge of the wall 7350. In some embodiments, the cylindrical tip and
annular gas orifice of the atomizing nozzle 7510 can be configured
to extend beyond the inside edge of the wall 7350.
[0269] In some embodiments, the vessel 710 also includes a gas
outlet tube 7340, which can pass through the center of the lid
7220, and extend into a solvent removal chamber 7230. In some
embodiments, the gas outlet tube 7340 can extend down from the lid
from about 15 to about 19 inches and can be about 1 inch in
diameter. In some embodiments, the gas outlet tube 7340 can extend
down from the lid from about 20 cm to about 60 cm and can be from
about 1 cm to about 4 cm in diameter. In some embodiments, a
conical fitting 7300 can be attached to the end of the gas outlet
tube 7340. In some embodiments, the conical fitting 7300 can be
used to narrow the diameter of the gas outlet tube 7340 at a gas
outlet 7310. In some embodiments, the gas outlet 7310 can be
approximately 0.5 inches in diameter. In some embodiments, the gas
outlet 7310 can be from about 0.5 cm to about 3 cm in diameter. In
some embodiments, a vortex stabilizer disk or ring 7360 can be
attached to the conical fitting 7300.
[0270] In some embodiments, a gas inlet pipe 7290 can be connected
to the wall 7350 of the solvent removal vessel 710 can be creating
a gas inlet 7280. In some embodiments, the solvent removal vessel
710 has a bottom 7250, which can include a product exit orifice
7260. In some embodiments, the exit orifice 7260 can be in the
center of the domed bottom 7250. In some embodiments, a product
outlet pipe 7270 can be attached to the exit orifice 7260. In some
embodiments, the solvent removal vessel 710, includes a temperature
control jacket (not shown). In some embodiments, temperature
controlling fluid can be circulated through a temperature control
jacket (not shown) of the solvent removal vessel 710, wherein the
temperature control jacket can surround the wall 7350 and the
bottom 7250 of the solvent removal vessel 710 for temperature
regulation in the solvent removal chamber 7230. In some
embodiments, the bottom 7250 of the solvent removal vessel 710
could be of a shape such as a dome, a cone or an angled flat
bottom.
[0271] In some embodiments, a carrier gas 7370 can be first heated
and humidified and then passed through a gas filtration system (not
shown) comprising a hydrophobic gas sterilizer (not shown). In some
embodiments, the carrier gas 7370 can be supplied to the solvent
removal vessel 710 through the gas inlet pipe 7290. In some
embodiments, the carrier gas 7370 can pass through the gas inlet
7280 and enter a gas rotation jet 7285. In some embodiments, the
gas rotation jet 7285 directs the flowing carrier gas 7370 inside
the solvent removal chamber 7230 such that the carrier gas 7370
exits the gas rotation jet 7285 (and enters the solvent removal
chamber 7230) horizontally and in a direction tangential to the
solvent removal vessel wall 5350. In some embodiments,
substantially all the carrier gas 7370 entering the solvent removal
chamber 7230 through the gas rotation jet 7285 can exit the solvent
removal chamber 7230 through the gas outlet 7310 and gas outlet
pipe 7340.
[0272] In some embodiments, the carrier gas 7370 supplied to
solvent removal chamber 7230 can be injected tangential to the wall
7350, causing the carrier gas 7370 to first travel slowly clockwise
(as viewed from above) around the solvent removal vessel 710 near
the wall 7350, and forming a slow gas rotation 7240. In some
embodiments, the solvent removal chamber 7230 can be pressured to
approximately 1 psig, creating a pressure differential between the
solvent removal chamber 7230 and the gas outlet pipe 7340. In some
embodiments, the carrier gas 7370 traveling in the solvent removal
chamber 7230 can be pulled inwards, as it circulates, towards the
gas outlet 7310. In some embodiments, the change in angular
momentum on the carrier gas 7370 causes the carrier gas 7370 to
accelerate as it moves closer to the gas outlet 7310. In some
embodiments, the acceleration of the carrier gas 7370 near the gas
outlet 7310 can be sufficiently strong to create an intense gas
vortex 7245 underneath the vortex stabilizer 7360 and gas outlet
7310. In some embodiments, the carrier gas 7370 can be pushed out
of the solvent removal chamber 7230 through the gas outlet 7310
into the gas outlet pipe 7340 for disposal after spinning through
the gas vortex 7245.
[0273] In some embodiments, the three-fluid atomizing nozzle 7510
can be supplied with a first fluid 7115, a second fluid 7120, and a
third fluid 7140, as described above. In a typical embodiment, the
first fluid can be a first component, the second fluid can be a
buffer solution, and the third fluid can be a gas. In some
embodiments, the resultant atomized droplets 7155, comprised of a
first fluid core and a second fluid shell, can be sprayed into the
solvent removal chamber 7230. In some embodiments, the atomized
droplets 7155 come into contact with the carrier gas 7370 being
circulated in the slow gas rotation 7240 as the atomized droplets
7155 can travel down through the solvent removal chamber 7230. In
some embodiments, the atomized droplets 7155 can be picked up by,
and incorporated in, the slow gas rotation 7240 and begin to
circulate through the solvent removal chamber 7230, and begin to
settle toward the domed bottom 7250.
[0274] In some embodiments, the solvent can be substantially
evaporated from the core of the atomized droplets 7155 as the
atomized droplets 7155 circulate along with the carrier gas 7370 in
the slow gas rotation 7240. In some embodiments, the evaporated
solvent can be removed from the solvent removal chamber 7230 along
with the circulating carrier gas 7370 through the gas outlet 7310
and into the gas outlet pipe 7340 for disposal. In some
embodiments, the gas outlet pipe 7340 can be equipped with a
sterile barrier filtration system (not shown) outside of solvent
removal vessel 710. The filtration system in one embodiment can be
comprised of filter (e.g. a course filter or conventional cyclone
separator) and a HEPA filter or other sterilizing gas filter.
[0275] In some embodiments, a portion of the atomized droplets
7155, traveling in the slow gas rotation 7240, can reach the
intense gas vortex 7245, where they can be kicked outwards again by
the intense centrifugal forces within the intense gas vortex 7245,
and thus are not removed through the gas outlet 7310. For example,
some of the atomized droplets 7155 kicked outwards by the gas
vortex 7245 can travel again through the slow gas rotation 7240 and
some of the atomized droplets begin to fall towards the domed
bottom 7250. In one embodiment, very few of the atomized droplets
7155 escape with the carrier gas 7370 through the gas outlet 7310.
In some embodiments, removal of substantially all the solvent from
the atomized droplets 7155 can afford large diameter synthetic
membrane vesicles droplets 7380 still coated in a fluid shell. In
some embodiments, droplets 7380 permanently fall out of the gas
vortex primarily to the domed bottom 7250 of the solvent removal
vessel 710. In some embodiments, the plurality of large diameter
synthetic membrane vesicles droplets 7380 form a suspension 7390 of
large diameter synthetic membrane vesicles particles in a solution
at the domed bottom 7250. In a typical embodiment, the large
diameter synthetic membrane vesicles are multivesicular
liposomes.
[0276] The solvent removal vessel 710 can optionally be equipped
with a two-fluid rinse nozzle 7400 in the lid 7220 of the solvent
removal vessel 710. In some embodiments, the two-fluid rinse nozzle
7400 can be positioned in the lid 7220 at 90 degrees clockwise from
the atomizing nozzle 7510. In some embodiments, the two-fluid rinse
nozzle 7400 can be positioned in the lid 7220 at 180 degrees
clockwise from the atomizing nozzle 7510. The rinse nozzle 7400 can
be fed a wall rinse solution 7410 and a filtered gas 7420 in order
to spray atomized wall rinse solution droplets 7415 into the
solvent removal vessel 710 for the purpose of rinsing any wayward
atomized emulsion droplets from the wall 7350 and rinsing the large
diameter synthetic membrane vesicles suspension 7390 to the product
exit orifice 7260. In a typical embodiment, the large diameter
synthetic membrane vesicles are multivesicular liposomes.
[0277] In some embodiments, solvent removal vessel 710 can be
optionally equipped with a lid-protecting gas inlet 7430 in the lid
7220. In some embodiments, solvent removal vessel 710 can be
optionally equipped with a lid-protecting gas inlet 7430 in the lid
7220 on a side opposite the atomizing nozzle 7510. In some
embodiments, humidified, sterilized, and filtered gas 7440 can be
streamed into the solvent removal chamber 7230 in a direction
tangential to the solvent removal vessel wall 7350 through the
lid-protecting gas inlet 7430, near the top of the solvent removal
chamber 7230. In some embodiments, gas 7440 can travel circularly,
as a lid protection jet, around the top of solvent removal chamber
7230 and, in essence, acts a gas cushion above the gas vortex 7245
and slow gas rotation 7240 to prevent droplet buildup on the lid
7220. In some embodiments, the gas streamed through the
lid-protecting gas inlet can be, for example, nitrogen gas (or a
nitrogen gas/aqueous vapor mixture) or air scrubbed of CO.sub.2. In
some embodiments, the carrier gas can be nitrogen gas (or a
nitrogen gas/aqueous vapor mixture) or air scrubbed of
CO.sub.2.
[0278] In some embodiments, the vortex stabilizer 7360 can be a
metal disk or ring extending radially outward from the conical
fitting 7300 and can be fitted flush with the gas outlet 7310 at
the end of the conical fitting 7300. In some embodiments, the
diameter of the vortex stabilizer 7360 is twice to six times the
diameter of the gas outlet 7310. In some embodiments, the vortex
stabilizer 7360 acts to ensure helical stability and integrity of
the intense gas vortex 7245 by protecting the tip of the intense
gas vortex 7245 from the turbulence caused by the spray emanating
from the atomizing nozzle 7510. In some embodiments, the stability
of the gas vortex 7245 can be maintained by placement of the
atomizing nozzle 7510 at a distance between 1/4 lid radius from the
wall 7350 and 1/4 lid radius from the gas outlet pipe 7340. In some
embodiments, the atomizing nozzle 7510 can be positioned at a
distance of 7/11 the radius of the lid 7220 from the gas outlet
pipe 7340. Strategic placement of the atomizing nozzle 7510
minimizes the impact on the intense gas vortex 7245 of spraying the
atomized droplets 7155 into the solvent removal chamber 7230 and
also minimizes the number of the atomized droplets 7155 that
impinge on the wall 7350. In some embodiments, the rinse nozzle
7400 placement is not critical as long as a substantial quantity of
rinse solution droplets 7415 enter the slow gas rotation 7240
within the solvent removal vessel 7240 and deposit on the vessel
walls 7350 and bottom 7250. In some embodiments, the rinse nozzle
7400 can also be of a one fluid (liquid) design fed by pressurized
rinse solution. In some emodiments, the nozzle can be 180 degrees
from the three fluid nozzle to suppress off access gas
rotation.
[0279] In some embodiments, the suspension 7390 collected at the
domed bottom 7250 of the solvent removal vessel 710 can be drained
and optionally pumped by a pump (not shown) from the domed bottom
7250 through the product exit orifice 7260 into the product outlet
pipe 7270 to optionally be further processed. In some embodiments,
the suspension 7390 can be further processed by a buffer exchange
through a series of diafilters.
[0280] In some embodiments, the solvent removal vessel of FIG. 7
can be a component of FIGS. 1A and B (component 50). In some
embodiments, the atomizing nozzle 7510 can receive fluid from the
line 2180 of FIG. 2. In some embodiments, the large diameter
synthetic membrane vesicles suspension 7390 can exit the solvent
removal vessel of FIG. 7 through the product exit orifice 7260 and
can enter the systems of FIG. 1C (through line 120), FIG. 8
(through line 8120), FIG. 10 (through line 0120) and/or FIG. 11
(through line 1120). Alternatively, the large diameter synthetic
membrane vesicles suspension 7390 can be collected directly from
the product exit orifice 7260.
[0281] In some embodiments, the apparatus is configured where the
gas inlet can be situated in the chamber to cause the gas to rotate
within the chamber around an axis of the chamber and a small
diameter gas exit orifice situated on that same axis and adapted to
form a small diameter, in relation to the tank diameter, intense
rapidly rotating gas vortex capable of substantially preventing
atomized droplets from exiting through the gas exit.
[0282] In some embodiments, the apparatus is configured where the
gas inlet, tank dimensions and gas exit diameter and position are
together adapted to retain the atomized droplets in the circulating
gas stream rather that force them to a wall. In some embodiments,
the atomized droplets can build up in the circulating gas stream
until the rate of settling to the liquid exit equals the rate of
generation by the atomizing nozzle.
[0283] Some embodiments provide an evaporation apparatus,
comprising a sealed solvent removal vessel, comprised of a lid, a
bottom, and a circular wall, at least one atomizing nozzle which is
not located on the central axis of the circular wall, a carrier gas
entrance orifice tangential to the circular wall, a carrier gas
exit orifice located on the central axis of the circular wall and
directed along the axis with a diameter of less than 1/5 of the
circular wall diameter, and a product exit orifice in the bottom of
the vessel. Additional embodiments include the processes for using
such a device and the MVL products made by the same. In some
embodiments, at least part of the vessel can be jacketed. In some
embodiments, the atomizing nozzle can be mounted to and extending
through the lid of the solvent removal vessel. In some embodiments,
the apparatus further comprises a rinse nozzle mounted to and
extending through the lid of the solvent removal vessel. In some
embodiments, the atomizing nozzle can be used to cause the gas
within the vessel to rotate. In some embodiments, the atomizing
nozzle can be angled at least 5 degrees measured off the central
axis of the wall and in a plane parallel to the wall nearest to it.
In some embodiments, the gas entrance orifice can be combined with
the atomizing nozzle. In some embodiments, the carrier gas exit
orifice can comprise a tube extending approximately 2/3 of the way
into the solvent removal vessel. In some embodiments, the tube can
be fitted with a narrowing cone and an optional annular ring. In
some embodiments, the gas exit orifice diameter can be less than a
1/10 diameter of the diameter of the inside of the tank. In some
embodiments, the atomizing nozzle can be a nozzle as disclosed
herein. In some embodiments, the ratio of the inside diameter of
the solvent removal vessel to the diameter of the carrier gas exit
orifice can be between approximately 5:1 and 100:1. In some
embodiments, the ratio of the inside diameter of the solvent
removal vessel to the diameter of the carrier gas exit orifice can
be between approximately 20:1 and 60:1. In some embodiments, the
solvent removal vessel is comprised of two, three, or four
atomizing nozzles which are used to spray the atomized droplets
into the solvent removal vessel.
FIG. 8
[0284] In some embodiments, the large diameter synthetic membrane
vesicles suspension resulting from the spray evaporation process
may optionally undergo a filtration and/or concentration process in
a continuous-flow particle-concentration system to concentrate the
large diameter synthetic membrane vesicles and remove the buffer
solution. One embodiment is a continuous-flow
particle-concentration unit, as well as the process to concentrate
and/or filter the large diameter synthetic membrane vesicles
particles so produced. In a typical embodiment, the large diameter
synthetic membrane vesicles are multivesicular liposomes. Other
embodiments of particle-concentration systems include the use of
one or more hydro-cyclones or one or more centrifuges. An example
of a hydro-cyclone is the Minicyclone or Minicyclone Array
available from ChemIndustrial Systems, Inc. of Cedarburg, Wis. An
example of a disk centrifuge is Model No. Pathfinder SC1-06-177
manufactured by GEA Westfalia Separator of Oelde, Germany.
[0285] In this document the term concentration unit, concentration
apparatus, concentration system, particle-concentration system,
particle-concentrating device, and particle concentrator are meant
to encompass units and processes that remove some or all of the
particle suspending medium of a particle suspension and therefore
concentrate the particles. Furthermore, the definition of these
terms encompasses the exchange of the suspending medium with a new
suspending medium, performed in one step or incrementally. These
two processes are closely related as exchanging the suspending
medium can be accomplished by concentrating the suspension and
adding new suspending medium. These terms relate to concentrating
the particle suspension and exchanging the suspending medium done
separately or simultaneously.
[0286] An embodiment of the present application is a system for
manufacturing formulations including a continuous-flow
diafiltration system, the process of using the system, and the
large diameter synthetic membrane vesicles made by the process. An
example of a continuous-flow diafiltration system is presented in
FIG. 8 and described herein. FIG. 8 is a schematic of one example
of a continuous-flow diafiltration system 810. In some embodiments,
the permeate discarded in the steps described herein contains a
buffer solution in which large diameter synthetic membrane vesicles
can be suspended as the large diameter synthetic membrane vesicles
suspension exits the solvent removal vessel. In a typical
embodiment, the large diameter synthetic membrane vesicles are
multivesicular liposomes.
[0287] In some embodiments, the large diameter synthetic membrane
vesicles suspension can be pumped from the solvent removal vessel,
such as the example vessel depicted in FIG. 7 and described above,
to the diafiltration system 810 through a particle suspension inlet
line 8120 (also seen in FIG. 1A, component 120 and in FIG. 7,
component 7270), reaching a first retentate vessel 8100. In some
embodiments, for each 1 L of large diameter synthetic membrane
vesicles suspension fed to the first retentate vessel 8100, a 2 L
solution can be fed into the first retentate vessel 8100 through a
line 8130, the solution first passing through a manual valve 8212
and a sterilizing hydrophilic filter 8170. In some embodiments, the
large diameter synthetic membrane vesicles are multivesicular
liposomes and the solution is a saline solution.
[0288] In some embodiments, a portion of the large diameter
synthetic membrane vesicles suspension in the first retentate
vessel 8100 can be pumped by pump 8110 through a cross-flow
(tangential-flow) filtration module 8150. For example, this can be
a hollow fiber type module. In one embodiment, the hollow fiber
filter used is Model No. CFP-2-E-8A (0.2 micron) manufactured by
Amersham Biosciences of Westborough, Mass. In some embodiments, the
pore size of the cross-flow (tangential-flow) filtration modules
8150, 8152 and 8154, can be chosen to retain the large diameter
synthetic membrane vesicles while allowing the suspending medium to
pass through the filter membranes as permeate. The pumps used in
the diafiltration system can be of various types, such as
peristaltic or rotary lobe positive displacement pumps. In some
embodiments, cross-flow recirculation pumps 8110, 8112 and 8114
operate with at least twice the permeate flow rate of their
associated cross-flow filter module and preferably 3 times, 5 times
or 10 times the permeate flow rates. In some embodiments, the
permeate can be drawn off through a permeate line 8160 (passing
through a sterilizing hydrophilic filter 8190 and a manual valve
8202), wherein for each 1 L of large diameter synthetic membrane
vesicles suspension added to the first retentate vessel 8100, 2.25
L of permeate can be removed and discarded. In some embodiments,
the retentate from the filtration module 8150 can be circulated
back into the first retentate vessel 8100 via a retentate line
8140. In some embodiments, for each 1 L of large diameter synthetic
membrane vesicles suspension added to the first retentate vessel
8100, a 0.75 L flow of the concentrated large diameter synthetic
membrane vesicles suspension can be removed from the first
retentate vessel 8100 through a feed line 8122 and a metering pump
8123 to be further filtered in a second retentate vessel 8200. In
some embodiments, the large diameter synthetic membrane vesicles
suspension exiting the first retentate vessel 8100 can be
concentrated by a factor of 1.33 (large diameter synthetic membrane
vesicles concentration is increased by 33%). In a typical
embodiment, the large diameter synthetic membrane vesicles are
multivesicular liposomes.
[0289] In some embodiments, similar filtration can occur in the
second retentate vessel 8200 as described above for the first
retentate vessel 8100. In some embodiments, the large diameter
synthetic membrane vesicles suspension can enter the second
retentate vessel 8200 at a rate of 0.75 L per 1 L added to the
first retentate vessel 8100. In some embodiments, a solution can be
fed into the second retentate vessel 8200 through a line 8132, the
solution first passing through a manual valve 8204 and a
sterilizing hydrophilic filter 8172, at a rate of 2 L per 0.75 L
concentrated large diameter synthetic membrane vesicles suspension
added to the second retentate vessel 8200. In some embodiments, a
portion of the large diameter synthetic membrane vesicles
suspension in the second retentate vessel 8200 can be pumped by
pump 8112 through a cross-flow (tangential-flow) filtration module
8152. In some embodiments, the permeate can be drawn off through a
permeate line 8162 (passing through a sterilizing hydrophilic
filter 8192 and a manual valve 8206), wherein for each 0.75 L of
large diameter synthetic membrane vesicles suspension added to the
second retentate vessel 8200, 2.25 L of permeate can be removed and
discarded. In some embodiments, the retentate from the filtration
module 8152 can be circulated back into the second retentate vessel
8200 via a retentate line 8142. In some embodiments, for each 0.75
L of large diameter synthetic membrane vesicles suspension added to
the second retentate vessel 8200, a 0.50 L flow of the concentrated
large diameter synthetic membrane vesicles suspension can be
removed from the second retentate vessel 8200 through a feed line
8124 and a metering pump 8125 to be further filtered in a final
retentate vessel 8300. In some embodiments, the large diameter
synthetic membrane vesicles suspension exiting the second retentate
vessel 8200 can now be concentrated 200%, whereas the buffer
solution concentration in the large diameter synthetic membrane
vesicles suspension can be roughly 9.1% with respect to the large
diameter synthetic membrane vesicles suspension in the inlet line
8120.
[0290] In some embodiments, similar filtration can occur in the
final retentate vessel 8300 as described above for the first
retentate vessel 8100 and second retentate vessel 8200. In some
embodiments, the concentrated large diameter synthetic membrane
vesicles suspension traveling through the feed line 8124 can enter
the final product vessel 8300 where it is further filtered and
concentrated to contain all the large diameter synthetic membrane
vesicles entering through the inlet line 8120, in the final product
vessel 8300. In some embodiments, a solution can be fed into the
final retentate vessel 8300 through a line 8132, the solution first
passing through a manual valve 8208 and a sterilizing hydrophilic
filter 8174, at a rate of 1.75 L per 0.50 L concentrated large
diameter synthetic membrane vesicles suspension added to the final
retentate vessel 8300. In some embodiments, the large diameter
synthetic membrane vesicles suspension can be pumped through the
pump 8114 to the cross-flow filtration module 8154, wherein the
permeate can be drawn off to be discarded through a permeate line
8164 (passing through a sterilizing hydrophilic filter 8194 and a
manual valve 8210) at a rate of 2.25 L per 0.5 L of large diameter
synthetic membrane vesicles suspension fed to the final product
vessel 8300. In some embodiments, in the final product vessel 8300,
the large diameter synthetic membrane vesicles suspension can
contain a buffer concentration. In some embodiments, the buffer
concentration can be as low as 2%.
[0291] Filters 8170, 8190, 8172, 8192, 8174, and 8194 are
sterilizing hydrophilic filters. Filters 8180, 8182, and 8184 are
sterilizing hydrophobic gas vent filters used in the retentate
vessels and fed by gas lines 8131, 8133, and 8135, respectively. In
some embodiments, the systems depicted in FIG. 1A through FIG. 8
can be operated in a sterile (aseptic) fashion). For example, the
addition of appropriate steam lines, condensate drain lines and
valves, can allow the system to be sterilized. In some embodiments,
all inputs and outputs are equipped with sterile barrier filters.
In a typical embodiment, the large diameter synthetic membrane
vesicles are multivesicular liposomes.
[0292] In some embodiments, a continuous-flow particle
concentration system can be configured where a set of cascading
particle suspension concentrators can be in series together adapted
to remove and/or replace the suspending medium of a particle
suspension in a continuous fashion. In some embodiments, the
cascading particle suspension concentrators can be cross flow
filters (FIG. 8, components 8150, 8152, 8154) and the suspending
medium can be replaced by diafiltration. In some embodiments, the
cascading particle suspension concentrators can be hydro-cyclones
or disk centrifuge units.
FIG. 9
[0293] FIG. 9A provides cross-sectional views of an atomized
droplet 902, and a large diameter synthetic membrane vesicles
particle 912. In a typical embodiment, the large diameter synthetic
membrane vesicles particle 912 can be formed by removal of an
organic solvent from the atomized droplet 902. In a typical
embodiment, the atomized droplet 902 can comprise a first component
core and a buffer solution shell 904. In some embodiments, first
component core can comprise a continuous phase 908 and a suspension
of droplets 906 in the continuous phase 908. In some embodiments,
the droplets 906 can be aqueous phase droplets and can be
surrounded by a continuous phase 908 that can be an organic
solvent. In some embodiments, the aqueous phase droplets can be
from about 10 nm to about 10.mu. in diameter. In some embodiments,
the aqueous phase droplets can be from about 500 nm to about 5.mu.
in diameter. In a typical embodiment, the aqueous phase droplets
can be from about 100 nm to about 2.mu. in diameter. For example
the average diameter of the aqueous phase droplets can be about
1.mu. in diameter. In some embodiments, the organic solvent can
then be removed in the solvent removal chamber to provide droplets
of large diameter synthetic membrane vesicles 912 within a shell of
dextrose/lysine 914. In a typical embodiment, the large diameter
synthetic membrane vesicles droplet 912 are a multivesicular
liposomes droplet. Multivesicular liposomes (MVL) are uniquely
different from other lipid-based drug delivery systems.
Topologically, MVL are defined as liposomes containing multiple
non-concentric chambers 916 within each droplet 912, resembling a
"foam-like" matrix. The chambers 916 of the MVL can have the same
volume as the first component particles, (e.g. 1.mu.) as shown in
FIG. 9A. The presence of internal membranes distributed as a
network throughout the MVL may serve to confer increased mechanical
strength to the vesicle, while still maintaining a high
volume:lipid ratio. Thus, both structurally and functionally the
MVL are unusual, novel and distinct from all other types of
liposomes.
[0294] In one embodiment, a large diameter synthetic membrane
vesicles droplet 930 made by the instant processes can be, as
depicted in FIG. 9B, for example, an atomized droplet 922
containing equal volumes of dextrose/lysine 924 and first
component, where the diameter of the atomized droplet is
approximately 39.7 .mu.m. In a typical embodiment, the large
diameter synthetic membrane vesicles droplet 930 can be a
multivesicular liposomes droplet. In this embodiment, the first
component core then is approximately 31.5 .mu.m and the
dextrose/lysine shell 924 can be approximately 4.1 .mu.m thick.
When the organic solvent is removed from this atomized droplet 922,
the resultant MVL 930 is 25 .mu.m in diameter (the dextrose/lysine
shell 924 is omitted for clarity). In some embodiments, chambers
936 of the MVL 930 can have the same volume as the suspension of
droplets in the continuous phase of the first component.
[0295] FIG. 10 is a schematic of one example of a continuous-flow
diafiltration system 1010 comprising a continuous flow centrifuge.
The large diameter synthetic membrane vesicles suspension can be
pumped from a solvent removal vessel to the continuous-flow
diafiltration system 1010.
[0296] In some embodiments, the continuous-flow diafiltration
system 1010 can include a solvent removal vessel as depicted in
FIG. 7 and as in component 70 of FIG. 1A and FIG. 1B. The large
diameter synthetic membrane vesicles suspension can travel to the
diafiltration system 1010 through a particle suspension inlet line
10120 (also seen in FIG. 7, component 7270), reaching a first
retentate vessel 10100. In some embodiments, for each 1 L of large
diameter synthetic membrane vesicles suspension fed to the first
retentate vessel 10100, a 2 L solution can be fed into the first
retentate vessel 10100 through a line 10130, the solution first
passing through a manual valve 10600 and a sterilizing hydrophilic
filter 10170. In some embodiments, the large diameter synthetic
membrane vesicles are multivesicular liposomes and the solution is
a saline solution.
[0297] In some embodiments, a portion of the large diameter
synthetic membrane vesicles suspension in the first retentate
vessel 10100 can be pumped by pump 10110 through a cross-flow
(tangential-flow) filtration module 10150. For example, this can be
a hollow fiber type module. In one embodiment, the hollow fiber
filter used is Model No. CFP-2-E-10A (0.2 micron) manufactured by
Amersham Biosciences of Westborough, Mass. In some embodiments, the
pore size of the cross-flow (tangential-flow) filtration modules
10150, 10152 and 10154, can be chosen to retain the large diameter
synthetic membrane vesicles while allowing the suspending medium to
pass through the filter membranes as permeate. The pumps used in
the diafiltration system can be of various types, such as
peristaltic or rotary lobe positive displacement pumps. In some
embodiments, cross-flow recirculation pumps 10110, 10112 and 10114
operate with at least twice the permeate flow rate of their
associated cross-flow filter module and preferably 3 times, 5 times
or 10 times the permeate flow rates. In some embodiments, the
permeate can be drawn off through a permeate line 10160 (passing
through a sterilizing hydrophilic filter 10190 and a manual valve
10602), wherein for each 1 L of large diameter synthetic membrane
vesicles suspension added to the first retentate vessel 10100, 2.25
L of permeate can be removed and discarded. In some embodiments,
the retentate from the filtration module 10150 can be circulated
back into the first retentate vessel 10100 via a retentate line
10140. In some embodiments, for each 1 L of large diameter
synthetic membrane vesicles suspension added to the first retentate
vessel 10100, a 0.75 L flow of the concentrated large diameter
synthetic membrane vesicles suspension can be removed from the
first retentate vessel 10100 through a feed line 10122 and a
metering pump 10123 to be further filtered in a second retentate
vessel 10200. In some embodiments, the large diameter synthetic
membrane vesicles suspension exiting the first retentate vessel
10100 can be concentrated by a factor of 1.33 (large diameter
synthetic membrane vesicles concentration is increased by 33%). In
a typical embodiment, the large diameter synthetic membrane
vesicles are multivesicular liposomes.
[0298] In some embodiments, similar filtration can occur in the
second retentate vessel 10200 as described above for the first
retentate vessel 10100. In some embodiments, the large diameter
synthetic membrane vesicles suspension can enter the second
retentate vessel 10200 at a rate of 0.75 L per 1 L added to the
first retentate vessel 10100. In some embodiments, a solution can
be fed into the second retentate vessel 10200 through a line 10132,
the solution first passing through a manual valve 10204 and a
sterilizing hydrophilic filter 10172, at a rate of 2 L per 0.75 L
concentrated large diameter synthetic membrane vesicles suspension
added to the second retentate vessel 10200. In some embodiments, a
portion of the large diameter synthetic membrane vesicles
suspension in the second retentate vessel 10200 can be pumped by
pump 10112 through a cross-flow (tangential-flow) filtration module
10152. In some embodiments, the permeate can be drawn off through a
permeate line 10162 (passing through a sterilizing hydrophilic
filter 10192 and a manual valve 10606), wherein for each 0.75 L of
large diameter synthetic membrane vesicles suspension added to the
second retentate vessel 10200, 2.25 L of permeate can be removed
and discarded. In some embodiments, the retentate from the
filtration module 10152 can be circulated back into the second
retentate vessel 10200 via a retentate line 10142. In some
embodiments, for each 0.75 L of large diameter synthetic membrane
vesicles suspension added to the second retentate vessel 10200, a
0.50 L flow of the concentrated large diameter synthetic membrane
vesicles suspension can be removed from the second retentate vessel
10200 through a feed line 10124 and a metering pump 10125 to be
further filtered in a third retentate vessel 10300. In some
embodiments, the large diameter synthetic membrane vesicles
suspension exiting the second retentate vessel 10200 can now be
concentrated 200%, whereas the buffer solution concentration in the
large diameter synthetic membrane vesicles suspension is roughly
9.1% with respect to the large diameter synthetic membrane vesicles
suspension in the inlet line 10120.
[0299] In some embodiments, similar filtration can occur in the
third retentate vessel 10300 as described above for the first
retentate vessel 10100 and second retentate vessel 10200. In some
embodiments, the concentrated large diameter synthetic membrane
vesicles suspension traveling through the feed line 10124 can enter
the third product vessel 10300 where it is further filtered and
concentrated to contain all the large diameter synthetic membrane
vesicles entering through the inlet line 10120, in the third
product vessel 10300. In some embodiments, a solution can be fed
into the third retentate vessel 10300 through a line 10132, the
solution first passing through a manual valve 102010 and a
sterilizing hydrophilic filter 10174, at a rate of 1.75 L per 0.50
L concentrated large diameter synthetic membrane vesicles
suspension added to the third retentate vessel 10300. In some
embodiments, the large diameter synthetic membrane vesicles
suspension can be pumped through the pump 10114 to the cross-flow
filtration module 10154, wherein the permeate can be drawn off to
be discarded through a permeate line 10164 (passing through a
sterilizing hydrophilic filter 10194 and a manual valve 10610) at a
rate of 2.25 L per 0.5 L of large diameter synthetic membrane
vesicles suspension fed to the third product vessel 10300. In some
embodiments, in the third product vessel 10300, the large diameter
synthetic membrane vesicles suspension can contain a buffer
concentration. In some embodiments, the buffer concentration can be
as low as 2%.
[0300] In some embodiments, the large diameter synthetic membrane
vesicles suspension traveling through a feed line 10126, can be
metered by a pump 10127 and can enter a continuous flow centrifuge
module 10400 wherein the supernantant can be pumped to be discarded
through a pump 10116 through a permeate line 10166 (passing through
a sterilizing hydrophilic filter 10196). In a typical embodiment,
the large diameter synthetic membrane vesicles are multivesicular
liposomes.
[0301] In some embodiments, the large diameter synthetic membrane
vesicles suspension traveling through a feed line 10128, can be
metered by a pump 10129 and can enter a final retentate vessel
10500. In a typical embodiment, the large diameter synthetic
membrane vesicles are multivesicular liposomes.
[0302] Filters 10170, 10190, 10172, 10192, 10174, 10194, and 10196
are sterilizing hydrophilic filters. Filters 10180, 10182, 10184
and 10186 are sterilizing hydrophobic gas vent filters used in the
retentate vessels and fed by gas lines 10131, 10133, 10135 and
10137, respectively.
[0303] FIG. 11 is a schematic of one example of a continuous-flow
centrifuge system 1110 comprising a plurality of continuous flow
centrifuges. The large diameter synthetic membrane vesicles
suspension can be pumped from a solvent removal vessel to the
continuous-flow centrifuge system 1110.
[0304] In some embodiments, the continuous-flow centrifuge system
1110 can include the solvent removal vessel as depicted in FIG. 7.
The large diameter synthetic membrane vesicles suspension can
travel to the centrifuge system 1110 through a particle suspension
inlet line 11120 (also seen in FIG. 7, component 7270, and as
represented by components 70 in FIG. 1a and FIG. 1B), reaching a
first retentate vessel 11100. In some embodiments, a large diameter
synthetic membrane vesicles suspension can be fed to the first
retentate vessel 11100, and a solution can be fed into the first
retentate vessel 11100 through a line 11130, the solution first
passing through a manual valve 11600 and a sterilizing hydrophilic
filter 11170. The large diameter synthetic membrane vesicles
suspension in the first retentate vessel 11100 can be pumped by a
pump 11220 to a first centrifuge module 11150. In some embodiments,
the permeate can be drawn off from the first centrifuge 11150
through a permeate line 11160 (passing through a sterilizing
hydrophilic filter 11190 being pumped by a pump 11110), and
discarded. In some embodiments, large diameter synthetic membrane
vesicles suspension can exit the first centrifuge module 11150
through a feed line 11122, metered by a pump 11123, to be further
processed in a second retentate vessel 11200. In some embodiments,
the large diameter synthetic membrane vesicles suspension flowing
in feed line 11122 can be concentrated by at least 33%. In a
typical embodiment, the large diameter synthetic membrane vesicles
are multivesicular liposomes and the solution is a saline
solution.
[0305] In some embodiments, the concentrated large diameter
synthetic membrane vesicles suspension can enter the second
retentate vessel 11200 for further processing, and a solution can
be fed through a line 11132, the solution first passing through a
manual valve 11604 and a sterilizing hydrophilic filter 11172. The
large diameter synthetic membrane vesicles suspension from the
second retentate vessel 11200 can be pumped by a pump 11240 to a
second centrifuge module 11152. In some embodiments, the permeate
can be drawn off from the second centrifuge 11152 through a
permeate line 11162 (passing through a sterilizing hydrophilic
filter 11192 being pumped by a pump 11120), and discarded. In some
embodiments, large diameter synthetic membrane vesicles suspension
can exit the second centrifuge module 11152 through a feed line
11124, metered by a pump 11125, to be further processed in a third
retentate vessel 11300. In some embodiments, the large diameter
synthetic membrane vesicles suspension flowing in feed line 11125
can be concentrated by at least 33%. In a typical embodiment, the
large diameter synthetic membrane vesicles are multivesicular
liposomes and the solution is a saline solution.
[0306] In some embodiments, the large diameter synthetic membrane
vesicles suspension traveling through the feed line 11124, can be
metered by a pump 11125 and can enter the third retentate vessel
11300 where it can be further processed. In some embodiments, a
solution can be fed into the third retentate vessel 11300 through a
line 11134, the solution first passing through a manual valve 11608
and a sterilizing hydrophilic filter 11174. In some embodiments,
the large diameter synthetic membrane vesicles suspension can be
pumped by a pump 11260 to a third centrifuge module 11154. In some
embodiments, the permeate can be drawn off from the centrifuge
through a permeate line 11164 (passing through a sterilizing
hydrophilic filter 11194 being pumped by pump 11130), and
discarded. In a typical embodiment, the large diameter synthetic
membrane vesicles are multivesicular liposomes.
[0307] In some embodiments, the concentrated large diameter
synthetic membrane vesicles suspension traveling through feed line
11166, can be metered by pump 11127 and can enter a final retentate
vessel 11400. In a typical embodiment, the large diameter synthetic
membrane vesicles are multivesicular liposomes.
[0308] Filters 11170, 11190, 11172, 11192, 11174, and 11194 are
sterilizing hydrophilic filters. Filters 11180, 11182, 11184 and
11186 are sterilizing hydrophobic gas vent filters used in the
retentate vessels and fed by gas lines 11131, 11133, 11135 and
11137, respectively. In a typical embodiment, the large diameter
synthetic membrane vesicles are multivesicular liposomes.
[0309] The systems depicted in FIG. 1A through FIG. 11 can be
operated in a sterile (aseptic) fashion. With the addition of
appropriate steam lines, condensate drain lines and valves, the
system can be sterilized. All inputs and outputs are equipped with
sterile barrier filters.
[0310] In some embodiments, the large diameter synthetic membrane
vesicles are multivesicular liposomes. In some embodiments, the
multivesicular liposomes further comprise bupivaciane, DEPC, DPPG,
and tricaprylin. In some embodiments, the multivesicular liposomes
further comprise bupivacaine phosphate, DEPC, DPPG, and
tricaprylin. In some embodiments, the multivesicular liposomes
further comprise bupivacaine, DEPC, DPPG, tricaprylin and
cholesterol. In some embodiments, the multivesicular liposomes
further comprise bupivacaine phosphate, DEPC, DPPG, tricaprylin and
cholesterol.
[0311] In some embodiments, the multivesicular liposomes further
comprise bupivaciane, dextrose, L-Lysine, DEPC, DPPG, and
tricaprylin. In some embodiments, the multivesicular liposomes
further comprise bupivacaine phosphate, dextrose, L-Lysine, DEPC,
DPPG, and tricaprylin. In some embodiments, the multivesicular
liposomes further comprise bupivacaine, dextrose, L-Lysine, DEPC,
DPPG, tricaprylin and cholesterol. In some embodiments, the
multivesicular liposomes further comprise bupivacaine phosphate,
dextrose, L-Lysine, DEPC, DPPG, tricaprylin and cholesterol.
[0312] In some embodiments, the multivesicular liposomes further
comprise bupivaciane, dextrose, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, dextrose, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine, dextrose, DEPC, DPPG, tricaprylin and cholesterol. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, dextrose, DEPC, DPPG, tricaprylin and
cholesterol.
[0313] In some embodiments, the multivesicular liposomes further
comprise bupivaciane, L-Lysine, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, L-Lysine, DEPC, DPPG, and tricaprylin. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine, L-Lysine, DEPC, DPPG, tricaprylin and cholesterol. In
some embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, L-Lysine, DEPC, DPPG, tricaprylin and
cholesterol.
[0314] In some embodiments, the multivesicular liposomes further
comprise bupivacaine, morphine, cytarabine, or their
pharmaceutically acceptable salts as the therapeutic agent. In some
embodiments, the multivesicular liposomes further comprise
bupivacaine phosphate, morphine sulfate, or cytarabine HCl.
[0315] In another embodiment, any one of the above described
embodiments can be used alone or in combination with any one or
more of the above described embodiments. For example, any above
described atomizing nozzle, evaporation apparatus, continuous-flow
emulsification system, continuous-flow diafiltration system,
continuous-flow diafiltration further comprising one or more
centrifuges, continuous-flow centrifuge system, or continuous
processing system can be used alone or in combination. Thus, an
evaporation apparatus can be used in conjunction with a three-fluid
atomizing nozzle. This evaporation system/atomizing nozzle can be
used with a continuous-flow emulsification system, as depicted in
FIGS. 1A, 1B, and 1C. The three-fluid atomizing nozzle/evaporation
apparatus combination can be used in conjunction with a
continuous-flow system, as depicted in FIGS. 8, 10, and 11. Any of
these combinations can be used to make multivesicular liposomes. In
particular any of the combinations can be used to make
multivesicular liposomes containing bupivacaine or its salts as the
therapeutic agent.
[0316] The following examples are meant to further illustrate the
embodiments, they are not meant to be limiting in any way.
Example 1
[0317] The following is an example utilizing the process parameters
and steps of the devices depicted in the Figures. The three fluids
applied to the atomizing nozzle (FIG. 1A and FIG. 1B, component 75;
FIG. 3A, component 310; FIG. 7, component 7510) as part of the
process of forming multivesicular liposomes have the following
compositions per liter.
[0318] The first fluid (FIG. 3A-3L, component 3115; FIG. 5,
component 5115; FIG. 7, component 7115) was a first liquid made up
of the first component, the first component having two components:
an organic phase and a first aqueous phase which are emulsified
with equal volumes. The organic phase was composed of
1,2-dierucoyl-sn-glycero-3-phosphocholine (17.78 g),
1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (1.056 g), cholesterol
(10.34 g), tricaprylin (4.32 g), water (0.70 g) and methylene
chloride (quantity sufficient to make 1 L total volume of the
organic phase. The first aqueous phase was composed of 0.2 molar
(200 mM) phosphoric acid and bupivacaine (40 g) and water (quantity
sufficient to make 1 L total volume of the first aqueous
phase).
[0319] The second fluid (FIG. 3A-3L, component 3120; FIG. 5,
component 5120; FIG. 7, component 7120) was a second liquid made up
of a second aqueous phase composed of L-lysine (monohydrate) (16.8
g), dextrose (13.25 g), and water (quantity sufficient to make 1 L
total volume of the second fluid.
[0320] The third fluid (FIG. 3A-3L, component 3140; FIG. 5,
component 5140; FIG. 7, component 7140) was nitrogen gas containing
water vapor (100% relative humidity at 42.degree. C. and 25
psig).
Preparation of Solvent Evaporation Chamber
[0321] Nitrogen was supplied to the solvent removal vessel (FIG. 1A
AND FIG. 1B, component 50; FIG. 7 component 710) through two
solvent removal vessel gas inlet lines: FIG. 1A components 115 and
110; FIG. 7 components 7280 and 7430) the main carrier gas inlet
line (component 115 of FIG. 1A AND FIG. 1B; component 7280 of FIG.
7), or main rotation jet, was tangential to vessel wall and
approximately 40% up from vessel bottom, causing clockwise rotation
viewed from above; and the lid protection gas inlet (component 110
of FIG. 1A AND FIG. 1B; component 7430 of FIG. 7), or lid
protection jet, at the corner of 11d (component 7220 of FIG. 7) and
vessel wall (component 7350 of FIG. 7), tangential to the wall and
moving in the same rotational direction (component 7240 of FIG. 7).
The nitrogen entering these inlet lines was humidified to 100%
relative humidity at 42.degree. C. and at 25 psig (pounds per
square inch gauge). The main rotation jet supplies 335 L/min at
42.degree. C. of humidified nitrogen while the lid protection jet
flow was 25 L/min at 42.degree. C. of humidified nitrogen (the lid
protection jet keeps deposit buildup off the lid). The nitrogen was
humidified prior to entering the solvent removal vessel by passing
it through a heated tube-in-shell heat exchanger (component 90,
FIG. 1A AND FIG. 1B) that was coated with water (humidification
water) followed by an excess-liquid water removal chamber
(component 45, FIG. 1A AND FIG. 1B) and liquid bleed. The nitrogen
was humidified to reduce evaporation of water from the spray which
raised the osmolality of the suspending buffer.
[0322] In this example, the solvent removal vessel had a volume of
approximately 138 liters, the inside diameter was 56 cm, the walls
were 52 cm high and the dome at the bottom was 10 cm deep. Thus the
inside height of the solvent removal vessel was 62 cm from the lid
to the bottom of the domed bottom. The gas outlet tube (component
80, FIG. 1A AND FIG. 1B; component 7340, FIG. 7) (including the
conical fitting FIG. 7, component 300 and vortex stabilizer FIG. 7,
component 7360) extends 42.5 cm into the solvent removal vessel
down from 11d. The diameter of the gas outlet tube was 2.3 cm
(inside diameter) and the conical fitting tapers 20 degrees to a
1.5 cm inside diameter for the gas outlet orifice (FIG. 7,
component 7310). The vortex stabilizer attached to the end of the
conical fitting had a diameter of 2.5 cm. The main rotation
(carrier) gas inlet was tangential to the wall and 37 cm down from
the lid and had an inside diameter of 1.9 cm.
[0323] The three-fluid atomizing nozzle (FIG. 1A and FIG. 1B,
component 75; FIG. 3A, component 310; FIG. 7, component 7510), the
rinse nozzle (FIG. 1A AND FIG. 1B, component 105; FIG. 7, component
7400), and lid protection gas inlet (FIG. 1A, component 110; FIG.
7, component 7340) all extend through the lid of the solvent
removal vessel (FIG. 1A AND FIG. 1B, component 50; FIG. 7 component
710) and were each centered 10.2 cm from the vessel wall (FIG. 7,
component 7350). The lid protection gas inlet (FIG. 7, component
7430) had an inside diameter of 0.95 cm, extends through the lid of
the vessel (FIG. 7, component 7220), and faced parallel to the
inside of the lid in a direction to give clockwise (viewed down
from above) rotation of the gas. Starting with the three-fluid
atomizing nozzle (FIG. 1A and FIG. 1B, component 75; FIG. 3A,
component 310; FIG. 7, component 7510) as zero degrees, the main
carrier gas inlet line was located 52 degrees clockwise, the rinse
nozzle was 90 degrees clockwise, and the lid protection gas inlet
was 135 degrees clockwise with its outlet approximately 180 degrees
clockwise.
[0324] The sides (FIG. 7, component 7350) and bottom (FIG. 7,
component 7250) of the solvent removal vessel were jacketed at
24.1.degree. C. by connection to a circulating bath. The
temperature in the jacket was adjusted to approximately match the
steady state gas exit temperature. Such a match prevented
evaporation and drying of the multivesicular liposomes on the wall
or condensation of water that could rupture, through osmotic shock,
the multivesicular liposomes that are being formed in the
vessel.
Preparation of a First Component
[0325] The recirculation loop (FIG. 2, component 2125) connected to
the high-shear mixer (FIG. 1A AND FIG. 1B, component 25, FIG. 2,
component 2130) (Ross model HSM-703XS-20 Sanitary Inline High Shear
Mixer equipped with a 3'' diameter X-5 Series rotor/stator for
operation to 14,400 rpm. (11,300 feet/min. tip speed) with gap ring
#3) was primed with methylene chloride to ensure that all air was
removed from the high-shear mixer. The jacket of the heat exchanger
(FIG. 1A AND FIG. 1B, component 30; FIG. 2 component 2170) was
supplied with 5.degree. C. coolant (water+50% ethylene glycol)
(FIG. 1A, components 96 and 97; FIG. 2, components 2110 and 2105).
The mixer seal lubricant tank (FIG. 1A AND FIG. 1B, component 10),
filled with water, was also cooled with 5.degree. C. coolant
(water+50% ethylene glycol). The mixer was started at a setting of
30 Hz (approx. 7,200 rpm) causing a flow around the mixer loop
estimated at 21,000 mL/min and with an internal volume of 280 mL.
Thus, fluid in the loop went through the mixer blades and heat
exchanger an average of every 0.8 seconds.
[0326] After the high shear mixer was primed with methylene
chloride, the organic phase and first aqueous phase peristaltic
pumps (components 12 and 2, respectively, of FIG. 1A AND FIG. 1B)
were concurrently started. The organic phase was pumped at 33
mL/min and the first aqueous phase was also pumped at 33 mL/min.
The organic phase and first aqueous phase entered the high-shear
mixer starting the formation of the first component. As the first
component circulated around the high shear mixer recirculation
loop, a small fraction of the flow was forced through the first
component exit line (FIG. 2, component 2180; FIG. 5, component
5114) to the three-fluid atomizing nozzle (FIG. 1A AND FIG. 1B,
component 75; FIG. 3A-3L, component 310; FIG. 5, component 505;
FIG. 7, component 7510) at 66 mL/min (total of the two flow rates).
Consequently, the priming methylene chloride was soon flushed from
the mixer loop by this flow (4.2 min per loop volume of flush).
[0327] Concurrent with the starting of the organic phase and first
aqueous phase peristaltic pumps, the second aqueous phase
(connected to the three-fluid atomizing nozzle) and wall rinse (to
the rinse nozzle (FIG. 1A and FIG. 1B, component 105; FIG. 7,
component 7400)) peristaltic pumps (components 22 and 64,
respectively, of FIG. 1A and FIG. 1B) were started and pumped their
respective components at 66 mL/min each. The wall rinse solution
was 33.5 g of dextrose per liter of water. Nitrogen at 60 psig
(room temperature, not humidified) was supplied to these two
nozzles (FIG. 1A and FIG. 1B, components 75 and 105; FIG. 7,
components 7510 and 7400). The second aqueous phase flowed through
the three fluid nozzle at 66 mL/min. The three-fluid atomizing
nozzle had a nitrogen flow rate of 51 L/min @ 1 atm and the wall
rinse nozzle (manufactured by GEA Process Engineering of Columbia,
Md.) had a flow rate of 66 L/min @ 1 atm.
[0328] After exiting the three-fluid atomizing nozzle, the formed
atomized emulsion droplets came into contact with the carrier gas
(nitrogen) in the solvent removal chamber (FIG. 7, component 7230)
of the solvent removal vessel (FIG. 1A and FIG. 1B, component 50;
(FIG. 7, component 710). The carrier gas rotated inside the chamber
(FIG. 7, component 7240) as a small, rapidly rotating, intense gas
vortex (FIG. 7, component 7245) formed at the exit orifice (FIG. 7,
component 7310). This allowed the droplets to contact the gas for
an extended period of time in order to effectuate methylene
chloride evaporation and removal. After removal of the methylene
chloride, the formed multivesicular liposome suspension droplets
(FIG. 7, component 7380) were collected as a suspension of
multivesicular liposomes (FIG. 7, component 7390) at the bottom of
an evaporation vessel (FIG. 1A and FIG. 1B, component 50; FIG. 7,
component 7250). The multivesicular liposomes were collected in the
solvent removal vessel and then drained out of the drain port (FIG.
1A and FIG. 1B, component 130; FIG. 7, component 7270) through a
peristaltic positive displacement pump (FIG. 1A and FIG. 1B,
component 125), set to pump slightly faster (approximately 200
mL/min) than the suspension drains out of the bottom (approximately
165 mL/min), therefore the exit stream of multivesicular liposome
suspension were periodically interrupted by small segments of
chamber gas. This pump rate prevented any appreciable venting of
solvent vapors into the room and protects the multivesicular
liposome suspension from exposure to high velocity gas streams or
foaming.
[0329] At system equilibrium, the nitrogen exiting the solvent
removal vessel through the gas outlet (FIG. 1A and FIG. 1B,
component 80; FIG. 7, component 7310) was at a temperature of
approximately 21.5.degree. C. The solvent removal vessel jacket was
cooled to approximately 24.1.degree. C. The temperature of the
first component (part of which travels to the three-fluid atomizing
nozzle) leaving the heat-exchanger was 15.3.degree. C. After
traveling through heat exchanger, and while traveling back to the
high-shear mixer through the recirculation line, the temperature of
the first component was approximately 14.3.degree. C.
[0330] At equilibrium the system then ran continuously making
multivesicular liposomes for as long as the feed solutions and
nitrogen were supplied. Five hundred mL samples of multivesicular
liposome suspension were taken from the suspension outlet (FIG. 7,
component 7260) in the solvent removal vessel and diafiltered
(using a hollow fiber filter, Model No. CFP-2-E-8A from Amersham
Biosciences of Westborough, Mass.) with four volumes of normal
saline on a small scale batch diafiltration system (FIG. 1A and
FIG. 1B, component 70). Optionally, settling of the multivesicular
liposomes and excess liquid could be decanted to obtain the final
multivesicular liposomes in a chosen aqueous solution at a chosen
MVL concentration. Full continuous operation can be obtained by
connecting the solvent removal vessel outlet pump to the apparatus
of FIG. 8.
Example 2
Preparation of First Component
[0331] The recirculation loop connected to the high-shear mixer
(FIG. 1A and FIG. 1B, component 25; FIG. 2, component 2130) (Ross
Model HSM-703XS-20 Sanitary Inline High Shear Mixer equipped with a
3'' diameter X-5 Series rotor/stator for operation to 14,400 rpm.
(11,300 feet/min. tip speed) with gap ring #3) was primed with
methylene chloride to ensure that all air was removed from the
high-shear mixer. The jacket of the heat exchanger (FIG. 1A and
FIG. 1B, component 30; FIG. 2, component 2170) was supplied with
5.degree. C. coolant (water+50% ethylene glycol). The mixer seal
lubricant tank, filled with water, was also cooled with 5.degree.
C. coolant (water+50% ethylene glycol). The high-shear mixer was
started at a setting of 25 Hz (6,000 rpm), 30 Hz approx. (7,200
rpm) or 35 Hz (8,400 rpm).
[0332] After the high shear mixer was primed with methylene
chloride, the organic phase and first aqueous phase peristaltic
pumps (FIG. 1A and FIG. 1B, components 12 and 2, respectively) were
concurrently started. The organic phase was pumped at 33 mL/min and
the first aqueous phase was also pumped at 33 mL/min. Entry of the
organic phase and first aqueous phase begin formation of the first
component. As the first component circulates around the high shear
mixer recirculation loop (FIG. 2, component 2125), a small fraction
of the flow was forced through the first component exit line (FIG.
2, component 2180) to the three-fluid atomizing nozzle (FIG. 1A and
FIG. 1B, component 75; FIG. 3A; FIG. 4A; FIG. 77510) at 66 mL/min
(total of the two flow rates), the temperature of the emulsion
being forced through the first component exit line (FIG. 2,
component 2180) was from 16.5 to 21.1.degree. C. The flow of the
organic phase and first aqueous rapidly flushed the priming
methylene chloride from the mixer loop (4.2 min per loop volume of
flush).
TABLE-US-00001 TABLE 1 Organic Phase Components (per 2 L Total
Volume) cholesterol (Nippon) 20.8 g DEPC (Nippon) 36.0 g DPPG
(Lipoid) 1.89 g tricaprylin (NOF) 8.81 g Water for injection 0.49
mL Methylene chloride (EMD) 2,569 g
[0333] The components of the organic phase are shown in Table
1.
TABLE-US-00002 TABLE 2 First Aqueous Phase Components (per 2 L
Total Volume) Bupivacaine (BASF) 80 g 0.20M H.sub.3PO.sub.4 (2 L,
Mallinckrodt) 200 mM
[0334] The components of the first aqueous phase are shown in Table
2.
TABLE-US-00003 TABLE 3 Particle Sizing of Diluted Emulsion Samples
by laser light scatter analysis. PSD (.mu.m) (volume based) Batch
d.sub.10 d.sub.50 d.sub.90 Span Ross 30 Hz 0.5 0.9 1.4 1.0 Ross 35
Hz 0.5 0.8 1.3 1.0 Ross 25 Hz 0.6 1.1 1.7 1.0 Batch Lot D 0.855
1.151 1.506 0.566 Batch Lot E 0.720 1.110 1.530 0.730
[0335] The emulsion samples were diluted and analyzed using a light
scattering device (Horiba Instruments La-910) and the results are
shown in Table 1.
[0336] The second aqueous phase (connected to the three-fluid
atomizing nozzle) and wall rinse (to the rinse nozzle (FIG. 1A and
FIG. 1B, component 105; FIG. 7, component 7400)) peristaltic pumps
(components 22 and 64, respectively, of FIG. 1A and FIG. 1B) were
started when the organic phase and first aqueous phase peristaltic
pumps were started. The wall rinse solution having 33.5 g of
dextrose per liter of water was introduced to the evaporation
chamber at a flow rate of 66 mL/min. Nitrogen at 60 psig (room
temperature, not humidified) was supplied to the atomizing nozzle.
The second aqueous phase was introduced to the three fluid nozzle
at a flow rate of 66 mL/min. The three-fluid atomizing nozzle has a
nitrogen flow rate of 51 L/min @ 1 atm and the wall rinse nozzle
(manufactured by GEA Process Engineering of Columbia, Md.) has a
flow rate of 66 L/min @ 1 atm. The emulsion, second aqueous phase
and nitrogen gas were combined using the three fluid nozzle to
afford atomized droplets (FIG. 3, component 3155; FIG. 7, component
7155) which traveled in the evaporation chamber until the majority
of the methylene chloride was removed from the atomized droplets.
Removal of the methylene chloride produced multivesicular liposomes
(FIG. 7, component 7380) which form a suspension (FIG. 7, component
7390) at the bottom of the evaporation chamber (FIG. 7, component
7250).
TABLE-US-00004 TABLE 4 Second Aqueous Phase Components (per 5 L
Total Volume) L-Lysine monohydrate 84 g 50% Dextrose soln (B Braun)
132.5 mL Deionized water (to final volume) 5 L
[0337] The components of the second aqueous phase are shown in
Table 4.
[0338] After system equilibrated (10 minutes), 500 mL samples of
multivesicular liposomes suspension were collected. The Ross RPM
was then changed and allowed to equilibrate for 10 minutes before
collecting the next 500 mL sample. This was repeated for the last
RPM. Each 500 mL batch was diafiltered in a batch mode (using a
hollow fiber filter, Model No. CFP-6-D-9A from Amersham Biosciences
of Westborough, Mass.), for four volumes with normal saline on a
small scale batch diafiltration system (FIG. 1, component 70). The
recirculation pump was set to 5,700 ml/min, while saline addition
pump was set to 410 mL/min. The permeate valve was adjusted to keep
the liquid volume in the diafiltration system constant at 1,000 and
thus was also 410 mL/min, since the minimum working volume of this
system was approximately 550 mL. In the present example, the
processed multivesicular liposomes suspension was allowed to settle
at 5.degree. C. and then supernatant was decanted to achieve
approximately 15 mg of bupivacaine per mL in the final
multivesicular liposomes suspension.
TABLE-US-00005 TABLE 5 Wall Rinse Solution (per 5 L Total Volume)
50% Dextrose soln (B Braun) 335 mL Deionized water (to final
volume) 5 L
[0339] As can be seen in FIG. 12, controlled release of bupivacaine
from MVL particles was examined in vitro at 37.degree. C. in 0.5%
ovine serum albumin dissolved in 50 mM phosphate buffered saline
(pH7) showing a similar release of bupivacaine to that in Batch Lot
A and B made by the process disclosed in patent WO 99/25319.
[0340] As can be seen in FIG. 13, the PK profile in rats of the
continuous process samples shows a similar sustained release
profile of bupivacaine to that in Batch Lot C made by the batch
process disclosed in patent WO 99/25319.
TABLE-US-00006 TABLE 6 Final Material Properties Total Free
Bupi.sup.i % Bupi.sup.i % PSD.sup.iii (.mu.m) pH Total Lipids Batch
(mg/mL) PPV.sup.ii (mg/mL) Free d10 d50 d90 Int Ext Chol.sup.iv
DEPC.sup.v DPPG.sup.vi TC.sup.vii 30 Hz 14.9 41 0.79 3.2 18.0 44.7
102.8 5.7 7.0 3.24 5.98 0.23 1.50 Batch 35 Hz 13.5 38 0.81 3.7 14.2
36.3 83.2 6.0 7.3 3.70 6.72 0.31 1.61 Batch 25 Hz 12.8 38 0.83 4.0
14.8 32.4 69.4 5.9 7.4 3.30 6.15 0.22 1.51 Batch .sup.ibupivacaine;
.sup.iipacked particle volume; .sup.iiiparticle size distribution
by mass; .sup.ivcholesterol;
.sup.v1,2-dierucoyl-sn-glycero-3-phosphocholine;
.sup.vi1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol;
.sup.viitricaprylin
Example 3
Heat Treatment of MVL Suspension
[0341] The system of FIG. 1B was used with the humidified rotation
gas (N2) supplied by combination electric heater and tube-in shell
heat exchanger as described for FIG. 1A, component 90. The system
was equilibrated for 10 minutes and a 1,000 ml sample of MVL
suspension, exiting the drain port (FIG. 1B, component 130) of the
solvent removal vessel 50, was collected. The MVL sample was
divided into two samples of 500 mL each. The first 500 mL MVL
sample was heat treated as follows. The heat treatment was
performed by rapidly adding 750 mL of 100.degree. C. dextrose
solution to the first sample to raise the mixture temperature up to
approximately 63.degree. C. After 30 seconds, 1,750 mL of
+5.degree. C. saline was rapidly added to lower the temperature of
the mixture to near room temperature (35.degree. C. or below). The
sample volume was now 3,000 mL. The second 500 mL multivesicular
liposomes sample was not heat treated. The second sample was
diluted with the same volumes of dextrose solution (750 mL) and
saline (1,750 mL) as the first sample but the solutions were at
room temperature.
[0342] Each sample was diafiltered batch wise, with 4 volumes of
saline. These 3,000 ml samples were each concentrated to 1 liter
and then diafiltered in a batch mode (using a hollow fiber filter,
Model No. CFP-6-D-9A from Amersham Biosciences of Westborough,
Mass.), for four volumes with normal saline on a small scale batch
diafiltration system (FIG. 1B, component 70). The recirculation
pump (FIG. 8, component 8110, 8112, and 8114) was set to 5,700
mL/min, while saline addition pump (FIG. 8, component 8170, 8172,
and 8174) was set to 410 mL/min. The permeate valve (FIG. 8,
component 8202, 8206, and 8210) was adjusted to 410 mL/min to keep
the liquid volume in the system constant at 1,000 mL. Since the
minimum working volume of this system was approximately 550 ml,
which was too large to allow these small samples to be concentrated
to the target bupivacaine concentration of 15 mg/mL, they were
allowed to settle at +5.degree. C. and the supernatant was decanted
to achieve approximately 15 mg of bupivacaine per ml in the final
MVL suspensions.
The analytical results for the samples were as follows:
TABLE-US-00007 TABLE 7 d.sub.10 d.sub.50 d.sub.90 ppv bupi/ml Not
heat treated 14.9 52.8 107.7 56% 16.84 Heat treated 18.2 51.3 95.9
55% 18
TABLE-US-00008 TABLE 8 Chol DEPC DPPG Tricap Sample Total mg/ml Not
heat treated 4.97 8.50 0.39 2.11 Heat treated 7.07 12.35 0.58
3.00
[0343] As can be seen in the Tables 7 and 8 above, the heat
treatment did not significantly effect the particle size
distribution of the MVLs and had only small effects on the
bupivacaine (active agent) content and lipid composition of the
particles.
[0344] The heat treatment did have a surprising effect on the
accelerated stability (30.degree. C. stability) of these MVL
suspensions, as can be seen in FIG. 14. Accelerated stability at
30.degree. C. was greatly improved with heat treatment. A lower
slope means less bupivacaine release and longer stability. Both
samples were usable products when stored at +5.degree. C. but the
heat treated sample was projected to have a much longer shelf
life.
[0345] As can be seen in FIG. 15, The PK profile in rats (measured
substantially as per patent number WO 02/096368) also shows an
improvement with heat treatment. Both samples have acceptable PK
profiles. The heat treated sample gives longer lasting sustained
release with higher serum values at 48 and 72 hours. The not heat
treated sample was essentially zero at 72 hours. The heat treatment
surprisingly improved the accelerated stability of the MVLs and
also improved their in-vivo release profile.
Example 4
Effect of Lowered 1St Aqueous Osmolality
[0346] The osmolality of the 1st aqueous solution in the previous
examples was significantly above 300 mOsm/kg. If there was no
appreciable loss of bupivacaine or phosphoric acid or water
transport across the forming phospholipid membranes during the MVL
production process, the internal chambers of the resultant MVLs
will be filled with an aqueous solution with an osmolality at or
near that of the final saline (300 mOsm/kg) storage suspending
medium. If, on the other hand, the osmolality of the 1st aqueous
and resultant MVLs was lower than that of saline, the MVLs will
shrink slightly as the saline draws water out of the internal
chambers. This will compress the phospholipids making up the MVL
membranes and make them more stable and less bupivacaine
permeable.
[0347] Both the lipid-solvent (LC) and the bupivacaine-acid (1st
aqueous) were made up at 1/2 the concentration of normal (see below
for formulations). This gave a 1st aqueous solution with an
osmolality of 189 mOsm/kg which was expected to cause the MVLs to
shrink and compress their membranes when diafiltered into normal
saline of 300 mOsm/kg.
TABLE-US-00009 TABLE 9 FIG 2 #s FIG 1B #s (but with no sterile
filters) 2160 12 LC, Solvent-lipids pump 33 ml/min 2120 2 1st
aqueous, acid Bupi pump 33 ml/min 22 Dextrose-lysine pump, 3 fluid
noz 66 ml/min 64 Dextrose only pump, wall rinse 66 ml/min noz 120
Output rate out of spray chamber 165 ml/min 115 & 110
Temperature of water humidified 44.6 C. N2 feeding rotation jets 90
Temperature of N2 exiting electric 46 C. heater 90 40 Flow rate of
humidification water 9 ml/min into steam generator 33 Flow to both
rotation jets (N2 400 L/min before steam) 57 Flow to top rotation
jet 15 L/min Chamber Jacket supply 25 C. temperature Chamber exit
temperature 21.9 C. heat exchanger emulsion inlet supplied to 2150
& 2180 3 fluid nozzle 18.8 C. 2175 heat exchanger emulsion
outlet 17.2 C. 2110 Heat exchanger coolant supply 4 C. 11 3 Fluid
nozzle N2 pressure 60 psig 21 Wall rinse nozzle pressure 60 psig 13
3 fluid nozzle N2 flow 54 L/min 23 Wall rinse nozzle N2 flow 53
L/min 25 Ross Mixer always with gap ring Hz on #3 Run at 30 VFD
resulting in this blade rotation rate 7,200 RPM 2160 10 Solvent
solution (LC), 4 liters 4 liters cholesterol Nippon) 20.8 g DEPC
(Nippon) 36.0 g DPPG (Lipoid) 1.89 g tricaprylin (NOF) 8.81 g WFI
1.4 ml MeCl (EMD) 5,138 g 2120 5 1st aqueous soln. Acid & Bupi,
2 liters 2 liters (emulsified by Ross, fed to rotor center)
Dissolve Bupivacaine base (BASF) 40 g in 2 liters of 0.112M H3PO4
112 mM (Mallinckrodt) Osmolality 189 mOsm 60 Dextrose/lysine
solution 5 liters (fed to sheath of 3 fluid nozzle) L-Lysine
monohydrate 84 g 50% Dextrose soln (B Braun) 105 g DI water to
final volume of 5 L Osmolality 145 mOsm 66 Dextrose wall rinse
solution 5 liters 50% Dextrose soln (B Braun) 329 g DI water to
final volume of 5 L Osmolality 148 mOsm
[0348] Since only 1/2 the lipids and 1/2 the previous examples
bupivacaine are processed per minute but the Dextrose Lysine
(second aqueous) and Dextrose only (wall rinse) solutions are
pumped at the normal rate, the concentration of bupivacaine MVLs
out of the spray chamber (FIG. 1, component 50) will be 1/2 of that
seen in the previous examples. A larger 1,000 mL sample was
therefore taken for concentration and diafiltration as in the
previous example. This sample was not heat treated.
TABLE-US-00010 TABLE 10 Analysis pf the final decanted product MVLs
are as follows: Bupi mg/ml % free PPV % d10 d50 D90 15.21 1.1% 71%
9.3 22.5 55.5
[0349] The accelerated stability (30C) plot for this low osmolality
sample can be seen in FIG. 14. The low osmolality sample was
significantly more stable than the not heat treated sample but less
stable than the heat treated sample.
[0350] As seen in FIG. 15, the Rat in-vivo release profile of the
low osmolality sample can be the most desirable as it has the
lowest initial peak and the longest duration; highest blood
concentrations at 72 and 96 hours. In this process, starting with a
1.sup.st aqueous solution with an osmolality lower than the final
MVL suspending solution was unexpectedly found to both increase MVL
storage stability, reduce the initial in-vivo release peak and
prolong the duration of bupivacaine delivery.
[0351] As can be seen from Table 10, this example produced a
smaller particle size d50 than the previous examples. As can be
seen from FIG. 14, the low osmolality sample, which was not heat
treated, has a much better accelerated stability than the not heat
treated sample of the previous example.
Example 5
Solvent Exchange/Concentration
[0352] FIGS. 8, 10 and 11 depict example continuous buffer exchange
and MVL concentration systems (seen in FIG. 1A and FIG. 1B,
component 70) which are fed a MVL suspension from a solvent removal
vessel (seen in FIG. 1A and FIG. 1B, component 50) by a solvent
line (seen in FIG. 1A and FIG. 1B, component 120).
[0353] These systems take the continuous flow of MVL suspension
produced in a solvent removal vessel (seen in FIG. 1A and FIG. 1B,
component 50) and exchange the suspending buffer for another
suspending medium (e.g. normal saline) and at the same time
optionally concentrate the MVL suspension. In most cases it is
desired to produce slightly over concentrated MVL suspensions in a
retentate vessel (seen in FIG. 8, component 8300; FIG. 10,
component 10500; and FIG. 11, component 11400). The lot is analyzed
and sterilely diluted to exact concentration before filling into
vials aseptically. The final concentration of the MVL is obtained
by processing in two or more stages connected in series.
Additionally, processing results in any percent of original
suspending buffer removal.
[0354] These systems include medium supply source, such as a tank
and a particle concentrating device. The particle concentrating
device is a hollow fiber tangential flow filter (e.g. Model No.
CFP-6-D-9A from Amersham Biosciences of Westborough, Mass.) or a
continuous or semi-continuous centrifuge (Centritech Lab III or
CARR ViaFuge Pilot from Pneumatic Scale Angelus Corp., Clearwater,
Fla.) or any other device that separates the MVLs from the
suspending medium.
[0355] The following constraints apply to analogous components of
all the steady state stages of the continuous buffer exchange and
MVL concentration systems:
[0356] In a continuous manner, the first stage of FIG. 8 consisting
of components 8120, 8100, 8122, 8123, 8110, 8190, 8202, 8160, 8150,
8140, 8180, 8131, 8130, 8212 and 8170 will exchange the initial
suspending buffer with normal saline.
[0357] In reference to FIG. 8, at steady state where a constant
volume is in the tank, the input volume flows; MVL suspension in
8120 and saline in 8130 must equal the volume out flows; permeate
8160 and MVL suspension transferred to the next stage by metering
pump 8123. This is done by controlling/fine adjusting any of the
above 4 flow rates to keep the volume in the tank constant, e.g.
saline fine adjustment valve 8212.
[0358] At steady state the mass or number of MVLs is conserved and
thus the input rate of the number of MVLs, concentration times the
flow rate, from pipe 8120 must equal the MVL outflow from 8122,
again number concentration times flow rate. This means that if the
volume outflow rate in 8122 is lower than the input volume flow
rate in 8120, the concentration of MVLs in the tank 8100 and pipe
8122 rises until the MVL in matches the MVL out. This gives a MVL
concentration factor, output MVL concentration divided by the input
MVL concentration equal to the volume flow in MVL input pipe, 8120,
divided by the volume flow in the output pipe, 8122.
[0359] The original buffer is diluted by saline at each stage. This
dilution factor is the flow rate of buffer in to the tank, 8100,
divided by the total of the volume flow rate of buffer in pipe 8120
and the volume flow rate of saline in pipe 8212. The MVLs take up
appreciable volume and so the flow rate of buffer into the tank,
8100, is the volume flow rate in pipe 8120 times one minus the
volume percentage of that suspension that is MVLs, (PPV %, Packed
Particle Volume percentage).
[0360] Adhering to these constraints, the first 2 stages of FIG. 8,
containing tanks 8100 and 8200 will reach steady state. The smaller
the tanks, the faster the equilibration time and the lower the
total holdup in the system. For the systems described in Examples 2
and 3, a tank volume of from 0.25 liter to 10 liters and preferably
0.75 liters is appropriate.
[0361] The third stage in FIG. 8 contains the final retentate
vessel, 8300, and is large enough to hold one lot of product, e.g.
40 liters. As shown, the volume in final retentate vessel 8300
remains constant but the concentration is continuously rising as
there is no MVL out flow, until it reaches the desired final
concentration of MVLs. By choosing appropriate flow rates in pipes
8124, 8208 and 8164 this stage is alternatively run with a constant
MVL concentration and rising volume.
[0362] When starting these systems, the systems are either started
with each tank filled to the appropriate volume with saline or they
are started empty and filled with the MVL suspension input and
saline input but only start the output pump, e.g. 8123, and hollow
fiber recirculation pump, e.g. 8110 when the tank for that stage
fills to its desired volume. Likewise at the end of a lot, the MVL
input line is switched to saline which moves all MVLs to the
product tank, or the saline and MVL inputs and the hollow fiber
recirculation pump is stopped to allow each tank to empty into the
next until the lot is again all in the product vessel.
[0363] Flow rate and additional parameters for the system of FIG. 8
using the hollow fiber cartridge and filtration rates and MVL
output rate of the 25 Hz Process, 30 Hz Process, or 35 Hz Process
of Example 1 is given below:
TABLE-US-00011 TABLE 11 (flow rates are for fluid traveling through
the components from FIG. 8) concentration factor of 100% in 1st 2
stages (buffer exchange only) MVL suspension in Saline in MVL
suspension out permeate Stage (mL/min) (mL/min) (mL/min) (mL/min) 1
8120 165 8130 410 8122 165 8160 410 2 8122 165 8132 410 8124 165
8162 410 3 8124 165 8134 245 0 8164 410 Lysine Net tank volume part
conc. dilution % Lysine % Stage change out PPV % factor left
exchanged System 810 Input, 100% 20.0% 100% 8120 >>> 1
8100 0 100% 20.0% 0.2435 24.4% 75.6% 2 8200 0 100% 20.0% 0.2435
5.9% 94.1% 3 8300 0 rising rising 0.3501 2.1% 97.9%
[0364] The flow rates in Table 11 result in buffer exchange only.
The MVL PPV remains at 20% but the original buffer concentration is
reduced by 99.7% with its concentration in the product being 2.1%
of original. For comparison, a 4 volume batch diafiltration
exchange as used in the examples, reduces the buffer concentration
to 1.8% or 98.2% exchanged.
TABLE-US-00012 TABLE 12 (flow rates are for fluid traveling through
the components from FIG. 8) A total concentration factor of 200% in
1st 2 stages and buffer exchange MVL suspension in Saline in MVL
suspension out permeate Stage (mL/min) (mL/min) (mL/min) (mL/min) 1
8120 165 8130 369 8122 124 8160 410 2 8122 124 8132 396 8124 110
8162 410 3 8124 110 8134 300 0 8164 410 Stage Lysine Net tank
volume concentration dilution % Lysine % Stage change factor PPV %
factor left exchanged System 810 Input, 100% 20.0% 100% 8120
>>> 1 8100 0 133% 26.6% 0.2635 26.3% 73.7% 2 8200 0 150%
40.0% 0.1870 4.9% 95.1% 3 8300 0 rising rising 0.1798 0.9%
99.1%
[0365] The flow rates in Table 12 result in buffer 99.1% exchanged
down to 0.9% of the original concentration.
TABLE-US-00013 TABLE 13 (flow rates are for fluid traveling through
the components from FIG. 11) concentration factor of 300% in 3
stages and buffer exchange Product is collected in Tank 11400 with
volume rising at 114.3 mL/min MVL suspension in Saline MVL
suspension out Supernatant out Stage (mL/min) (mL/min) (mL/min)
(ml/min) 1 11120 165 11130 949 11122 114.3 11160 1,000 2 11122 114
11132 1000 11124 114.3 11162 1,000 3 11124 114 11134 1000 11166
114.3 11164 1,000 Stage Lysine Net tank volume concentration
dilution % Lysine % Stage change factor PPV % factor left exchanged
System 1110 Input, 100% 20.0% 100% 11120 >>> 1 11100 0
144% 28.9% 0.1221 12.2% 87.8% 2 11200 0 144% 41.7% 0.0752 0.9%
99.1% 3 1130 0 144% 60.1% 0.0625 0.1% 99.9%
[0366] The flow rates in Table 13 result in a concentration factor
of 300% with the same input conditions used for Tables 11 and
12.
[0367] Systems can be assembled with any combination of hollow
fiber cartridges and centrifuges. They can have two or three or
more stages. With more stages the buffer is exchanged with less
saline but there is more equipment to keep aseptic.
[0368] The semi-continuous centrifuges above, Centritech Lab III or
CARR ViaFuge Pilot, are operated in an aseptic fashion. The
semi-continuous centrifuges are capable of very high particle
concentration factors e.g. 100 to 1 and can also discharge MVL
concentrate up to 80% PPV %. The Centritech Lab III is
semi-continuous as it has a constant feed with an intermittent
concentrate discharge, every 10 seconds to 2 minutes. The ViaFuge
Pilot has a constant feed which is interrupted every 2 to 10
minutes by a rapid discharge cycle.
[0369] FIG. 10 depicts continuous buffer exchange and MVL
concentration systems where the hollow fiber filters operate at
lower particle concentrations, a condition where they have higher
permeate rates. The centrifuge (seen in FIG. 10, component 10400),
e.g. CARR ViaFuge Pilot, is used to do a final concentration into
the final product vessel (seen in FIG. 10, component 10500). In
this system all three stages including the tanks (seen in FIG. 10,
components 10100, 10200 and 10300) run at both constant volume and
MVL concentration while the final tank (seen in FIG. 10, component
10500) collects the concentrated buffer exchanged MVL
suspension.
[0370] FIG. 11 depicts a system for continuous buffer exchange and
MVL concentration including only centrifuges. Using typical process
rates for the ViaFuge Pilot and the same feed conditions as the
hollow fiber system, provides a system that exchanges 99.9% of the
buffer while concentrating the MVL suspension by a factor of 3 when
using the parameters from Table 13. The saline inputs should only
flow when the input MVL stream is flowing.
[0371] Any tanks connected to these centrifuges must be large
enough to accommodate the intermittent nature of the centrifuge
input and output.
[0372] While the present invention has been described with
reference to the specific embodiments thereof, it should be
understood by those skilled in the art that various changes may be
made and equivalents may be substituted without departing from the
true spirit and scope of the invention. In addition, many
modifications may be made to adapt a particular situation,
material, composition of matter, process, process step or steps, to
the objective, spirit and scope of the present invention. All such
modifications are intended to be within the scope of the claims
appended hereto.
* * * * *