U.S. patent application number 10/731224 was filed with the patent office on 2005-01-06 for compositions and methods of delivery of pharmacological agents.
This patent application is currently assigned to American BioScience, Inc.. Invention is credited to Ci, Sherry Xiaopei, De, Tapas, Desai, Neil P., Grim, Bridget Beals, Soon-Shiong, Patrick, Trieu, Vuong, Yang, Andrew, Yao, Oiang.
Application Number | 20050004002 10/731224 |
Document ID | / |
Family ID | 34891317 |
Filed Date | 2005-01-06 |
United States Patent
Application |
20050004002 |
Kind Code |
A1 |
Desai, Neil P. ; et
al. |
January 6, 2005 |
Compositions and methods of delivery of pharmacological agents
Abstract
The present invention relates to a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, which carrier comprises a protein, for example, human
serum albumin and/or deferoxamine. The human serum albumin is
present in an amount effective to reduce one or more side effects
associated with administration of the pharmaceutical composition.
The invention also provides methods for reducing one or more side
effects of administration of the pharmaceutical composition,
methods for inhibiting microbial growth and oxidation in the
pharmaceutical composition, and methods for enhancing transport and
binding of a pharmaceutical agent to a cell.
Inventors: |
Desai, Neil P.; (Santa
Monica, CA) ; Yang, Andrew; (Rosemead, CA) ;
De, Tapas; (Los Angeles, CA) ; Ci, Sherry
Xiaopei; (San Marino, CA) ; Soon-Shiong, Patrick;
(Los Angeles, CA) ; Trieu, Vuong; (Calabasas,
CA) ; Yao, Oiang; (Culver City, CA) ; Grim,
Bridget Beals; (Torrance, CA) |
Correspondence
Address: |
LEYDIG VOIT & MAYER, LTD
TWO PRUDENTIAL PLAZA, SUITE 4900
180 NORTH STETSON AVENUE
CHICAGO
IL
60601-6780
US
|
Assignee: |
American BioScience, Inc.
Santa Monica
CA
|
Family ID: |
34891317 |
Appl. No.: |
10/731224 |
Filed: |
December 9, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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60432317 |
Dec 9, 2002 |
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60526544 |
Dec 3, 2003 |
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60526773 |
Dec 4, 2003 |
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60527177 |
Dec 5, 2003 |
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Current U.S.
Class: |
514/15.2 ;
514/170; 514/171; 514/291; 514/34; 514/449; 514/553; 514/567;
514/731 |
Current CPC
Class: |
A61K 31/343 20130101;
A61P 37/06 20180101; A61K 31/165 20130101; A61K 31/436 20130101;
A61P 9/04 20180101; Y10S 977/705 20130101; A61P 29/00 20180101;
A61K 31/16 20130101; A61P 39/02 20180101; A61K 31/198 20130101;
A61P 19/02 20180101; B82Y 5/00 20130101; A61K 31/355 20130101; Y10S
977/906 20130101; A61K 47/42 20130101; Y10S 977/779 20130101; A61K
31/00 20130101; A61P 9/00 20180101; A61P 43/00 20180101; Y10S
977/773 20130101; A61K 31/05 20130101; A61P 35/00 20180101; A61K
31/7048 20130101; A61P 9/10 20180101; A61K 47/18 20130101; A61K
31/427 20130101; A61K 31/4745 20130101; A61K 9/146 20130101; A61P
23/00 20180101; A61K 31/24 20130101; A61K 31/337 20130101; Y10S
977/911 20130101; A61K 9/1075 20130101; A61K 9/0078 20130101; A61K
9/0019 20130101; A61K 9/19 20130101; A61K 38/13 20130101; A61K
9/107 20130101; A61K 31/164 20130101; A61P 31/04 20180101; A61K
31/05 20130101; A61K 2300/00 20130101; A61K 31/16 20130101; A61K
2300/00 20130101; A61K 31/337 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
514/002 ;
514/012; 514/011; 514/291; 514/170; 514/171; 514/449; 514/034;
514/553; 514/567; 514/731 |
International
Class: |
A61K 038/13; A61K
038/17; A61K 031/704; A61K 031/337; A61K 031/56; A61K 031/4745 |
Claims
What is claimed is:
1. A pharmaceutical composition comprising a pharmaceutical agent
and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises albumin in an amount
effective to reduce one or more side effects of administration of
the pharmaceutical composition into a human, and wherein the
pharmaceutically acceptable carrier comprises deferoxamine in an
amount effective to inhibit microbial growth in the pharmaceutical
composition.
2. The pharmaceutical composition of claim 1, wherein the
pharmaceutical agent is selected from the group consisting of
anticancer agents, anesthetics, antimicrotubule agents, agents to
treat cardiovascular disorders, antihypertensives,
anti-inflammatory agents, anti-arthritic agents, antiasthmatics,
analgesics, vasoactive agents, immunosuppressive agents, antifungal
agents, antiarrhythmic agents, antibiotics, and hormones.
3. The pharmaceutical composition of claim 2, wherein the
pharmaceutical agent is selected from the group consisting of
paclitaxel, docetaxel, taxanes, camptothecin, propofol, amiodarone,
cyclosporine, rapamycin, amphotericin, liothyronine, epothilones,
colchicines, thyroid hormones, vasoactive intestinal peptide,
corticosteroids, melatonin, tacrolimus, mycophenolic acids, and
derivatives thereof.
4. The pharmaceutical composition of claim 3, wherein the
pharmaceutical agent is propofol.
5. The pharmaceutical composition of claim 1, wherein the
pharmaceutical composition is a liquid and comprises from about
0.1% to about 25% by weight of albumin.
6. The pharmaceutical composition of claim 5, wherein the
pharmaceutical composition comprises about 0.5% to about 5% by
weight of albumin.
7. The pharmaceutical composition of claim 5, wherein the
pharmaceutical composition is dehydrated.
8. The pharmaceutical composition of claim 6, wherein the
pharmaceutical composition is lyophilized.
9. The pharmaceutical composition of claim 1, wherein the
pharmaceutical composition comprises a mesylate salt of
deferoxamine.
10. The pharmaceutical composition of claim 9, wherein the
pharmaceutical composition is a liquid and comprises from about
0.0001% to about 0.5% by weight of deferoxamine mesylate.
11. The pharmaceutical composition of claim 10, wherein the
pharmaceutical composition comprises about 0.1% by weight of
deferoxamine mesylate.
12. The pharmaceutical composition of claim 10, wherein the
pharmaceutical composition is dehydrated.
13. The pharmaceutical composition of claim 12, wherein the
pharmaceutical composition is lyophilized.
14. The pharmaceutical composition of claim 1, wherein the
pharmaceutical composition is an oil-in-water emulsion.
15. The pharmaceutical composition of claim 5, wherein the
pharmaceutical agent is propofol.
16. The pharmaceutical composition of claim 10, wherein the
pharmaceutical agent is propofol.
17. The pharmaceutical composition of claim 9, wherein the
pharmaceutical agent is propofol, the propofol is present in an
amount from about 0.1% to about 5% by weight, the albumin is
present in an amount from about 0.1% to about 25% by weight, and
the deferoxamine mesylate is present in an amount from about
0.0001% to about 0.5% by weight.
18. A pharmaceutical composition comprising a pharmaceutical agent
and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises albumin in an amount
effective to reduce one or more side effects of administration of
the pharmaceutical composition into a human, and wherein the
pharmaceutically acceptable carrier comprises deferoxamine in an
amount effective to inhibit oxidation in the pharmaceutical
composition.
19. A method for reducing one or more side effects associated with
administration of a pharmaceutical composition to a human, which
method comprises administering to a human a pharmaceutical
composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises albumin and deferoxamine.
20. The method of claim 19, wherein the pharmaceutical agent is
selected from the group consisting of anticancer agents,
anesthetics, antimicrotubule agents, agents to treat cardiovascular
disorders, antihypertensives, anti-inflammatory agents,
anti-arthritic agents, antiasthmatics, analgesics, vasoactive
agents, immunosuppressive agents, antifungal agents, antiarrhythmic
agents, antibiotics, and hormones.
21. The method of claim 20, wherein the pharmaceutical agent is
selected from the group consisting of paclitaxel, docetaxel,
taxanes, camptothecin, propofol, amiodarone, cyclosporine,
rapamycin, amphotericin, liothyronine, epothilones, colchicines,
thyroid hormones, vasoactive intestinal peptide, corticosteroids,
melatonin, tacrolimus, mycophenolic acids, and derivatives
thereof.
22. The method of claim 21, wherein the pharmaceutical agent is
propofol.
23. The method of claim 19, wherein the pharmaceutical composition
is a liquid and comprises from about 0.1% to about 25% by weight of
albumin.
24. The method of claim 23, wherein the pharmaceutical composition
comprises about 0.5% to about 5% by weight of albumin.
25. The method of claim 23, wherein the pharmaceutical composition
is dehydrated.
26. The method of claim 25, wherein the pharmaceutical composition
is lyophilized.
27. The method of claim 23, wherein the pharmaceutical agent is
propofol.
28. The method of claim 19, wherein the pharmaceutical composition
comprises a mesylate salt of deferoxamine.
29. The method of claim 28, wherein the pharmaceutical composition
is a liquid and comprises from about 0.0001% to about 0.5% by
weight of deferoxamine mesylate.
30. The method of claim 29, wherein the pharmaceutical composition
comprises about 0.1% by weight of deferoxamine mesylate.
31. The method of claim 29, wherein the pharmaceutical composition
is dehydrated.
32. The method of claim 31, wherein the pharmaceutical composition
is lyophilized.
33. The method of claim 29, wherein the pharmaceutical agent is
propofol.
34. The method of claim 28, wherein the pharmaceutical agent is
propofol, the propofol is present in an amount from about 0.1% to
about 5% by weight, the albumin is present in an amount from about
0.1% to about 25% by weight, and the deferoxamine mesylate is
present in an amount from about 0.0001% to about 0.5% by
weight.
35. The method of claim 19, wherein the pharmaceutical composition
is administered to the human via intravenous administration,
intra-arterial administration, intrapulmonary administration, oral
administration, inhalation, intra-tracheal administration,
intravesicular administration, intramuscular administration,
subcutaneous administration, intraocular administration,
intrathecal administration, or transdermal administration.
36. The method of claim 19, wherein the one or more side effects
are selected from the group consisting of myelosuppression,
neurotoxicity, hypersensitivity, venous irritation, inflammation,
phlebitis, pain, skin irritation, and combinations thereof.
37. A method for inhibiting microbial growth in a pharmaceutical
composition, which method comprises preparing a pharmaceutical
composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises deferoxamine in an amount effective
for inhibiting microbial growth in the pharmaceutical
composition.
38. The method of claim 37, wherein the pharmaceutical composition
comprises a mesylate salt of deferoxamine.
39. The method of claim 38, wherein the pharmaceutical composition
is a liquid and comprises from about 0.0001% to about 0.5% by
weight of deferoxamine mesylate.
40. The method of claim 39, wherein the pharmaceutical composition
comprises about 0.1% by weight of deferoxamine mesylate.
41. The method of claim 39, wherein the pharmaceutical composition
is dehydrated.
42. The method of claim 41, wherein the pharmaceutical composition
is lyophilized.
43. The method of claim 37, wherein the pharmaceutical composition
further comprises albumin.
44. A method for inhibiting oxidation of a pharmaceutical
composition, which method comprises preparing a pharmaceutical
composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises deferoxamine in an amount effective
for inhibiting oxidation of the pharmaceutical composition.
45. The method of claim 44, wherein the pharmaceutical composition
comprises a mesylate salt of deferoxamine.
46. The method of claim 45, wherein the pharmaceutical composition
is a liquid and comprises from about 0.0001% to about 0.5% by
weight of deferoxamine mesylate.
47. The method of claim 46, wherein the pharmaceutical composition
comprises about 0.1% by weight of deferoxamine mesylate.
48. The method of claim 46, wherein the pharmaceutical composition
is dehydrated.
49. The method of claim 48, wherein the pharmaceutical composition
is lyophilized.
50. The method of claim 44, wherein the pharmaceutical composition
further comprises albumin.
51. A method for enhancing transport of a pharmaceutical agent to
the site of an infirmity, which method comprises administering to a
human a pharmaceutical composition comprising a pharmaceutical
agent and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises albumin, and wherein
the ratio of albumin to pharmaceutical agent in the pharmaceutical
composition is about 18:1 or less.
52. The method of claim 51, wherein the pharmaceutical agent is
selected from the group consisting of anticancer agents,
anesthetics, antimicrotubule agents, agents to treat cardiovascular
disorders, antihypertensives, anti-inflammatory agents,
anti-arthritic agents, antiasthmatics, analgesics, vasoactive
agents, immunosuppressive agents, antifingal agents, antiarrhythmic
agents, antibiotics, and hormones.
53. The method of claim 52, wherein the pharmaceutical agent is
selected from the group consisting of paclitaxel, docetaxel,
taxanes, camptothecin, propofol, amiodarone, cyclosporine,
rapamycin, amphotericin, liothyronine, epothilones, colchicines,
thyroid hormones, vasoactive intestinal peptide, corticosteroids,
melatonin, tacrolimus, mycophenolic acids, and derivatives
thereof.
54. The method of claim 51, wherein the pharmaceutical agent is a
nucleic acid sequence.
55. The method of claim 54, wherein the nucleic acid sequence is a
DNA sequence
56. The method of claim 51, wherein the infirmity is selected from
the group consisting of cancer, arthritis, and cardiovascular
disease.
57. The method of claim 51, wherein the pharmaceutical composition
is a liquid and comprises from about 0.1% to about 25% by weight of
albumin.
58. The method of claim 57, wherein the pharmaceutical composition
comprises about 0.5% to about 5% by weight of albumin.
59. The method of claim 57, wherein the pharmaceutical composition
is dehydrated.
60. The method of claim 59, wherein the pharmaceutical composition
is lyophilized.
61. The method of claim 51, wherein the ratio of albumin to
pharmaceutical agent in the pharmaceutical composition is about
12:1 or less.
62. The method of claim 51, wherein the ratio of albumin to
pharmaceutical agent in the pharmaceutical composition is about 9:1
or less.
63. The method of claim 51, wherein the pharmaceutical composition
is administered to the human via intravenous administration,
intra-arterial administration, intrapulmonary administration, oral
administration, inhalation, intra-tracheal administration,
intravesicular administration, intramuscular administration,
subcutaneous administration, intraocular administration,
intrathecal administration, or transdermal administration.
64. A method for enhancing binding of a pharmaceutical agent to a
cell in vitro or in vivo, which method comprises administering to
said cell in vitro or in vivo, a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
albumin, and wherein the ratio of albumin to pharmaceutical agent
in the pharmaceutical composition is about 18:1 or less.
65. The method of claim 64, wherein the pharmaceutical agent is
selected from the group consisting of anticancer agents,
anesthetics, antimicrotubule agents, agents to treat cardiovascular
disorders, antihypertensives, anti-inflammatory agents,
anti-arthritic agents, antiasthmatics, analgesics, vasoactive
agents, immunosuppressive agents, antifungal agents, antiarrhythmic
agents, antibiotics, and hormones.
66. The method of claim 65, wherein the pharmaceutical agent is
selected from the group consisting of paclitaxel, docetaxel,
taxanes, camptothecin, propofol, amiodarone, cyclosporine,
rapamycin, amphotericin, liothyronine, epothilones, colchicines,
thyroid hormones, vasoactive intestinal peptide, corticosteroids,
melatonin, tacrolimus, mycophenolic acids, and derivatives
thereof.
67. The method of claim 64, wherein the pharmaceutical agent is a
nucleic acid sequence.
68. The method of claim 67, wherein the nucleic acid sequence is a
DNA sequence
69. The method of claim 64, wherein the cell is an endothelial
cell.
70. The method of claim 64, wherein the pharmaceutical composition
is a liquid and comprises from about 0.1% to about 25% by weight of
albumin.
71. The method of claim 70, wherein the pharmaceutical composition
comprises about 0.5% to about 5% by weight of albumin.
72. The method of claim 70, wherein the pharmaceutical composition
is dehydrated.
73. The method of claim 72, wherein the pharmaceutical composition
is lyophilized.
74. The method of claim 64, wherein the ratio of albumin to
pharmaceutical agent in the pharmaceutical composition is about
12:1 or less.
75. The method of claim 64, wherein the ratio of albumin to
pharmaceutical gent in the pharmaceutical composition is about 9:1
or less
76. The method of claim 64, wherein the pharmaceutical composition
is administered to the cell in vivo via intravenous administration,
intra-arterial administration, intrapulmonary administration, oral
administration, inhalation, intra-tracheal administration,
intravesicular administration, intramuscular administration,
subcutaneous administration, intraocular administration,
intrathecal administration, or transdermal administration.
77. A pharmaceutical composition comprising a pharmaceutical agent
and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises albumin in an amount
effective to reduce one or more side effects of administration of
the pharmaceutical composition into a human, and wherein the ratio
of albumin to pharmaceutical agent is about 18:1 or less.
78. The pharmaceutical composition of claim 77, wherein the ratio
of albumin to pharmaceutical agent in the pharmaceutical
composition is about 12:1 or less.
79. The pharmaceutical composition of claim 77, wherein the ratio
of albumin to pharmaceutical agent in the pharmaceutical
composition is about 9:1 or less.
80. A pharmaceutical composition comprising a pharmaceutical agent
and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises albumin in an amount
effective to increase transport of the drug to the site of
infirmity in a human, and wherein the ratio of albumin to
pharmaceutical agent is about 18:1 or less.
81. The pharmaceutical composition of claim 80, wherein the ratio
of albumin to pharmaceutical agent in the pharmaceutical
composition is about 12:1 or less.
82. The pharmaceutical composition of claim 80, wherein the ratio
of albumin to pharmaceutical agent in the pharmaceutical
composition is about 9:1 or less.
83. The pharmaceutical composition of claim 80, wherein the
infirmity is selected from the group consisting of cancer,
arthritis, and cardiovascular disease.
84. The pharmaceutical composition of claim 1, wherein the ratio of
albumin to pharmaceutical agent is about 18:1 or less.
85. A method for increasing the transport of a pharmaceutical agent
to a cell in vitro or in vivo by combining said agent with a
protein, wherein said protein binds a specific cell-surface
receptor on said cell, wherein said binding of the
protein-pharmaceutical agent combination with the said receptor
causes the transport to occur, and wherein the ratio of protein to
pharmaceutical agent is about 18:1 or less.
86. The method of claim 85, wherein the protein is albumin.
87. The method of claim 85, wherein the pharmaceutical agent is
selected from the group consisting of anticancer agents,
anesthetics, antimicrotubule agents, agents to treat cardiovascular
disorders, antihypertensives, anti-inflammatory agents,
anti-arthritic agents, antiasthmatics, analgesics, vasoactive
agents, immunosuppressive agents, antifungal agents, antiarrhythmic
agents, antibiotics, and hormones.
88. The method of claim 87, wherein the pharmaceutical agent is
selected from the group consisting of paclitaxel, docetaxel,
taxanes, camptothecin, propofol, amiodarone, cyclosporine,
rapamycin, amphotericin, liothyronine, epothilones, colchicines,
thyroid hormones, vasoactive intestinal peptide, corticosteroids,
melatonin, tacrolimus, mycophenolic acids, and derivatives
thereof.
89. The method of claim 85, wherein the ratio of albumin to
pharmaceutical agent in the pharmaceutical composition is about
12:1 or less.
90. The method of claim 85, wherein the ratio of albumin to
pharmaceutical agent in the pharmaceutical composition is about 9:1
or less.
91. A pharmaceutical composition comprising a pharmaceutical agent
and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises a protein in an
amount effective to reduce one or more side effects of
administration of the pharmaceutical composition into a human, and
wherein the ratio of protein to pharmaceutical agent is about 18:1
or less.
92. The pharmaceutical composition of claim 91, wherein the ratio
of protein to pharmaceutical agent in the pharmaceutical
composition is about 12:1 or less.
93. The pharmaceutical composition of claim 91, wherein the ratio
of protein to pharmaceutical agent in the pharmaceutical
composition is about 9:1 or less.
Description
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This patent application claims the benefit of U.S.
Provisional Patent Application No. 60/432,317 filed Dec. 9, 2002,
U.S. Provisional Patent Application (Attorney Docket No. 225519)
filed Dec. 3, 2003, U.S. Provisional Patent Application (Attorney
Docket No. 225549) filed Dec. 4, 2003, and U.S. Provisional Patent
Application (Attorney Docket No. 225585) filed Dec. 5, 2003.
FIELD OF THE INVENTION
[0002] This invention pertains to pharmaceutical compositions
comprising pharmaceutically active agents for parenteral or other
internal use, which have the effect of reducing certain undesirable
side effects upon administration when compared with available
formulations of similar drugs.
BACKGROUND OF THE INVENTION
[0003] It is well recognized that many drugs for parenteral use,
especially those administered intravenously, cause undesirable side
effects such as venous irritation, phlebitis, burning and pain on
injection, venous thrombosis, extravasation, and other
administration related side effects. Many of these drugs are
insoluble in water, and are thus formulated with solubilizing
agents, surfactants, solvents, and/or emulsifiers that are
irritating, allergenic, or toxic when administered to patients
(see, e.g., Briggs et al., Anesthesis 37, 1099 (1982), and Waugh et
al., Am. J. Hosp. Pharmacists, 48, 1520 (1991)). Often, the free
drug present in the formulation induces pain or irritation upon
administration. For example, phlebitis was observed in 50% of
patients who received peripheral vein administration of ifosfamide
and vinorelbine as first-line chemotherapy for advanced non-small
cell lung carcinoma. (see, e.g., Vallejo et al., Am. J. Clin.
Oncol., 19(6), 584-8 (1996)). Moreover, vancomycin has been shown
to induce side effects such as phlebitis (see, e.g., Lopes Rocha et
al., Braz. J. Infect. Dis., 6(4), 196-200 (2002)). The use of
cisplatin, gemcitabine, and SU5416 in patients with solid tumors
has resulted in adverse events such as deep venous thromboses and
phlebitis (see, e.g., Kuenen et al., J. Clin. Oncol., 20(6),
1657-67 (2002)). In addition, propofol, an anesthetic agent, can
induce pain on injection, burning and vein irritation, particularly
when administered as a lecithin-stabilized fat emulsion (see, e.g,
Tan et al., Anathesia, 53, 468-76, (1998)). Other drugs that
exhibit administration-associated side effects include, for
example, Taxol (paclitaxel) (see, e.g., package insert for Taxol
I.V.), codarone (amiodarone hydrochloride) (see, e.g., package
insert for Codarone I.V.), the thyroid hormone T3 or liothyronine
(commercially available as Triostat), thiotepa, bleomycin, and
diagnostic radiocontrast agents.
[0004] Another problem associated with the manufacture of drugs for
injection, particularly water insoluble drugs, is the assurance of
sterility. Sterile manufacturing of drug emulsions/dispersions can
be accomplished by absolute sterilization of all the components
before manufacture, followed by absolutely aseptic technique in all
stages of manufacture. However, such methods are time consuming and
expensive. In addition, the oxidation of drug formulations by
exposure to air during manufacture or storage can lead to, for
example, reduced pH, drug degradation, and discoloration, thereby
destabilizing the drug formulation and/or reducing shelf life.
[0005] To circumvent the problems associated with
administration-related side effects of drug formulations, alternate
formulations have been attempted. With respect to propofol, for
example, methods for reducing propofol-induced pain include
increasing the fat content of the solvent (e.g., long chain
triglycerides (LCT)), premedication, pretreatment with
non-steroidal drugs, local anaesthetics, opioids, the addition of
lidocaine, the addition of cyclodextrin, and microfiltration (see,
e.g., Mayer et al., Anaesthesist, 45(11), 1082-4 (1996), Davies, et
al. Anaesthesia, 57, 557-61 (2002), Doenicke, et al., Anaesth.
Analg., 82, 472-4 (1996), Larsen et al., Anaesthesitis 50, 842-5
(2001), Lilley et al., Anaesthesia, 51, 815-8 (1996), Bielen et
al., Anesth. Analg., 82(5), 920-4 (1996), and Knibbe et al., Br. J.
Clin. Pharmacol., 47(6), 653-60 (1999)). These formulations,
however, induce other side effects (e.g., cardiovascular
complications), or cause destabilization of propofol emulsions.
[0006] To overcome the problem of bacterial contamination, propofol
formulations have been developed with antibacterial agents, such as
an EDTA equivalent (e.g., edetate), pentetate, or
sulfite-containing agents, or they have been have been formulated
with a lower pH (see, e.g., U.S. Pat. Nos. 5,714,520, 5,731,355,
5,731,356, 6,028,108, 6,100,302, 6,147,122, 6,177,477, 6,399,087,
6,469,069, and International Patent Application No. WO 99/39696).
Since edetate and pentetate are metal ion chelators, however, they
have the potential to be dangerous by scavenging the essential
metal ions from the body system. Moreover, the addition of
sulphites to drug formulations presents potential adverse effects
to the pediatric population and for those in the general population
who are allergic to sulphur.
[0007] Thus, there remains a need for a composition and method that
reduce or eliminate the side effects associated with the parenteral
or in vivo administration of drugs. There also is a need for a
pharmaceutical composition that is sterile, and methods of
preparing such a composition. In addition, there is a need for a
pharmaceutical composition and method that reduce or eliminate
oxidation of pharmaceutical formulations to prevent drug
destabilization.
[0008] The invention provides such compositions and methods. These
and other advantages of the invention, as well as additional
inventive features, will be apparent from the description of the
invention provided herein.
BRIEF SUMMARY OF THE INVENTION
[0009] The invention provides various embodiments of pharmaceutical
compositions. One, some, or all of the properties of the various
embodiments can be found in different embodiments of the invention
and still fall within the scope of the appended claims.
[0010] The invention provides a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
a protein, such as albumin, more preferably human serum albumin, in
an amount effective to reduce one or more side effects of
administration of the pharmaceutical composition into a human, and
wherein the pharmaceutically acceptable carrier comprises
deferoxamine in an amount effective to inhibit microbial growth in
the pharmaceutical composition. The invention also provides a
pharmaceutical composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises a protein, such as albumin, in an
amount effective to reduce one or more side effects of
administration of the pharmaceutical composition into a human, and
wherein the pharmaceutically acceptable carrier comprises
deferoxamine in an amount effective to inhibit oxidation in the
pharmaceutical composition.
[0011] The invention provides a method for reducing one or more
side effects associated with administration of a pharmaceutical
composition to a human comprising (a) administering to a human a
pharmaceutical composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises albumin and deferoxamine. Also
provided are methods for inhibiting microbial growth, or for
inhibiting oxidation, or for inhibiting microbial growth and
oxidation in a pharmaceutical composition. These methods comprise
preparing a pharmaceutical composition comprising a pharmaceutical
agent and a pharmaceutically acceptable carrier, wherein the
pharmaceutically acceptable carrier comprises deferoxamine in an
amount effective for inhibiting microbial growth or in an amount
effective for inhibiting oxidation in the pharmaceutical
composition.
[0012] The invention also provides a method for enhancing transport
of a pharmaceutical agent to the site of an infirmity, which method
comprises administering to a human a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
albumin, and wherein the ratio of albumin to pharmaceutical agent
in the pharmaceutical composition is about 18:1 or less. The
invention further provides a method for enhancing binding of a
pharmaceutical agent to a cell in vitro or in vivo, which method
comprises administering to said cell in vitro or in vivo a
pharmaceutical composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises albumin, and wherein the ratio of
albumin to pharmaceutical agent in the pharmaceutical composition
is about 18:1 or less.
[0013] The invention also provides a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
albumin in an amount effective to increase transport of the drug to
the site of infirmity in a human, and wherein the ratio of albumin
to pharmaceutical agent is about 18:1 or less.
[0014] The invention further provides a method for increasing the
transport of a pharmaceutical agent to a cell in vitro or in vivo
by combining said agent with a protein, wherein said protein binds
a specific cell-surface receptor on said cell, wherein said binding
of the protein-pharmaceutical agent combination with the said
receptor causes the transport to occur, and wherein the ratio of
protein to pharmaceutical agent is about 18:1 or less.
DETAILED DESCRIPTION OF THE INVENTION
[0015] The invention provides a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
a protein such as albumin, preferably human serum albumin, in an
amount effective to reduce one or more side effects of
administration of the pharmaceutical composition to a human, and
wherein the pharmaceutically acceptable carrier comprises
deferoxamine in an amount effective to inhibit microbial growth in
the pharmaceutical composition. The invention also provides a
pharmaceutical composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises a protein such as albumin in an amount
effective to reduce one or more side effects of administration of
the pharmaceutical composition to a human, and wherein the
pharmaceutically acceptable carrier comprises deferoxamine in an
amount effective to inhibit oxidation in the pharmaceutical
composition.
[0016] Any suitable pharmaceutical agent can be used in the
inventive pharmaceutical composition. Suitable pharmaceutical
agents include, but are not limited to, anticancer agents or
antineoplastics, antimicrotubule agents, immunosuppressive agents,
anesthetics, hormones, agents for use in cardiovascular disorders,
antiarrythmics, antibiotics, antifungals, antihypertensives,
antiasthmatics, analgesics, anti-inflammatory agents,
anti-arthritic agents, and vasoactive agents. The invention is
useful with many other drug classes as well. More specifically,
suitable pharmaceutical agents include, but are not limited to,
taxanes, (e.g., Taxol.RTM. (paclitaxel), and Taxotere.TM.
(docetaxel)), epothilones, camptothecin, colchicine, amiodarone,
thyroid hormones, vasoactive peptides (e.g., vasoactive intestinal
peptide), amphotericin, corticosteroids, propofol, melatonin,
cyclosporine, rapamycin (sirolimus), tacrolimus, mycophenolic
acids, ifosfamide, vinorelbine, vancomycin, gemcitabine, SU5416,
thiotepa, bleomycin, diagnostic radiocontrast agents, and
derivatives thereof. Other drugs that are useful in the inventive
composition are described in, for example, U.S. Pat. No. 5,916,596
and co-pending U.S. patent application Ser. No. 09/446,783.
Preferably, the pharmaceutical agent is propofol, paclitaxel, or
docetaxel. More preferably, the pharmaceutical agent is propofol or
paclitaxel. Most preferably, the pharmaceutical agent is
propofol.
[0017] Taxol.RTM. (paclitaxel) (Bristol-Myers Squibb) is active
against carcinomas of the ovary, breast, lung, esophagus and head
and neck. Taxol, however, has been shown to induce toxicities
associated with administration, as well significant acute and
cumulative toxicity, such as myelosuppression, neutropenic fever,
anaphylactic reaction, and peripheral neuropathy. Because
paclitaxel is poorly soluble in water, cremophor typically is used
as a solvent, requiring large infusion volumes and special tubing
and filters. Cremophor is associated with side effects that can be
severe, including anaphylaxis and other hypersensitivity reactions
that can require pretreatment with corticosteroids, antihistamines,
and H.sub.2 blockers (see, e.g., Gelderblom et al., Eur. J. of
Cancer, 37, 1590-1598, (2001)). Taxotere.TM. (docetaxel) is used in
treatment of anthracycline-resistant breast cancer, but also has
previously been shown to induce side effects of hypersensitivity
and fluid retention that can be severe. Epothilone (and derivatives
thereof) also typically is administered in cremophor, and has been
shown to induce severe neutropenia, hypersensitivity, and
neuropathy.
[0018] Propofol (2,6-diisopropylphenol) is a hydrophobic,
water-insoluble oil, which is widely used as an intravenous
anesthetic agent to induce and maintain general anesthesia and
sedation of humans and animals. Propofol typically is administered
directly into the bloodstream and crosses the blood-brain barrier.
Pharmaceutical compositions comprising propofol must have
sufficient lipid solubility to cross this barrier and depress the
relevant mechanisms of the brain. Propofol has a maximum solubility
in water of 1.0+/-0.02 .mu.M at 22.5.degree. C. (see, e.g., Tonner
et al., Anesthesiology, 77, 926-931 (1992)). As such, propofol is
generally formulated as an emulsion containing solubilizing agents,
surfactants, solvents, or as an oil-in-water emulsion (see, e.g.,
U.S. Pat. Nos. 6,150,423, 6,326, 406, and 6,362,234). In addition
to the active pharmaceutical agent, the compositions of the present
invention include pharmaceutical carriers, or excipients. The
choice of carrier is not necessarily critical, and any of the
carriers known in the art can be used in the composition. The
choice of carrier is preferably determined, in part, by the
particular site to which the pharmaceutical composition is to be
administered and the particular method used to administer the
pharmaceutical composition. Preferably, the pharmaceutically
acceptable carrier comprises proteins. Any suitable protein can be
used. Examples of suitable proteins include, but are not limited to
albumin, immunoglobulins including IgA, lipoproteins,
apolipoprotein B, beta-2-macroglobulin, thyroglobulin and the like.
Most preferably, the pharmaceutically acceptable carrier comprises
albumin, most preferably human serum albumin. Proteins, including
albumin, suitable for the invention may be natural in origin or
synthetically prepared.
[0019] Human serum albumin (HSA) is a highly soluble globular
protein of M.sub.r 65K and consists of 585 amino acids. HSA is the
most abundant protein in the plasma and accounts for 70-80% of the
colloid osmotic pressure of human plasma. The amino acid sequence
of HSA contains a total of 17 disulphide bridges, one free thiol
(Cys 34), and a single tryptophan (Trp 214). Intravenous use of HSA
solution has been indicated for the prevention and treatment of
hypovolumic shock (see, e.g., Tullis, JAMA, 237, 355-360, 460-463,
(1977)) and Houser et al., Surgery, Gynecology and Obstetrics, 150,
811-816 (1980)) and in conjunction with exchange transfusion in the
treatment of neonatal hyperbilirubinemia (see, e.g., Finlayson,
Seminars in Thrombosis and Hemostasis, 6, 85-120, (1980)).
[0020] Human serum albumin (HSA) has multiple hydrophobic binding
sites (a total of eight for fatty acids, an endogenous ligand of
HSA) and binds a diverse set of drugs, especially neutral and
negatively charged hydrophobic compounds (Goodman et al., The
Pharmacological Basis of Therapeutics, 9.sup.th ed, McGraw-Hill New
York (1996)). Two high affinity binding sites have been proposed in
subdomains IIA and IIIA of HSA, which are highly elongated
hydrophobic pockets with charged lysine and arginine residues near
the surface which function as attachment points for polar ligand
features (see, e.g., Fehske et al., Biochem. Pharmcol., 30, 687-92
(1981), Vorum, Dan. Med. Bull., 46, 379-99 (1999), Kragh-Hansen,
Dan. Med. Bull., 1441, 131-40 (1990), Curry et al., Nat. Struct.
Biol., 5, 827-35 (1998), Sugio et al., Protein. Eng., 12, 439-46
(1999), He et al., Nature, 358, 209-15 (1992), and Carter et al.,
Adv. Protein. Chem., 45, 153-203 (1994)). Paclitaxel and propofol
have been shown to bind HSA (see, e.g., Paal et al., Eur. J.
Biochem., 268(7), 2187-91 (2001), Purcell et al., Biochim. Biophys.
Acta, 1478(1), 61-8 (2000), Altmayer et al., Arzneimittelforschung,
45, 1053-6 (1995), and Garrido et al., Rev. Esp. Anestestiol.
Reanim., 41, 308-12 (1994)). In addition, docetaxel has been shown
to bind to human plasma proteins (see, e.g., Urien et al., Invest.
New Drugs, 14(2), 147-51 (1996)). Thus, while not wishing to be
bound to any particular theory, it is believed that the inclusion
of proteins such as albumin in the inventive pharmaceutical
compositions results in a reduction in side effects associated with
administration of the pharmaceutical composition that is due, at
least in part, to the binding of human serum albumin to any free
drug that is present in the composition.
[0021] The amount of albumin included in the pharmaceutical
composition of the present invention will vary depending on the
pharmaceutical active agent, other excipients, and the route and
site of intended administration. Desirably, the amount of albumin
included in the composition is an amount effective to reduce one or
more side effects the active pharmaceutical agent due to the of
administration of the inventive pharmaceutical composition to a
human. Typically, the pharmaceutical composition is prepared in
liquid form, and the albumin is then added in solution. Preferably,
the pharmaceutical composition, in liquid form, comprises from
about 0.1% to about 25% by weight (e.g. about 0.5% by weight, about
5% by weight, about 10% by weight, about 15% by weight, or about
20% by weight) of albumin. Most preferably, the pharmaceutical
composition, in liquid form, comprises about 0.5% to about 5% by
weight of albumin. The pharmaceutical composition can be
dehydrated, for example, by lyophilization, spray-drying,
fluidized-bed drying, wet granulation, and other suitable methods
known in the art. When the composition is prepared in solid form,
such as by wet granulation, fluidized-bed drying, and other methods
known to those skilled in the art, the albumin preferably is
applied to the active pharmaceutical agent, and other excipients if
present, as a solution. The HSA solution preferably is from about
0.1% to about 25% by weight (about 0.5% by weight, about 5% by
weight, about 10% by weight, about 15% by weight, or about 20% by
weight) of albumin.
[0022] In addition to albumin, the compositions of the present
invention preferably comprise deferoxamine. Deferoxamine is a
natural product isolated from Streptomyces pilosus, and is capable
of forming iron complexes. Deferoxamine mesylate for injection USP,
for example, is approved by the Food and Drug Administration as an
iron-chelating agent and is available for intramuscular,
subcutaneous, and intravenous administration. Deferoxamine mesylate
USP is a white to off-white powder. It is freely soluble in water
and its molecular weight is 656.79. The chemical name for
deferoxamine mesylate is N-[5-[3-[(5-aminopentyl)-hydro-
xycarbamoyl]-propion-amido]pentyl]-3[[5-((N-hydroxyacetamido)pentyl]-carba-
moyl]propionohydroxamic acid monomethanesulfonate (salt), and its
structural formula is
C.sub.25H.sub.48N.sub.6O.sub.8.CH.sub.3SO.sub.3H. As described in
the Examples, deferoxamine, or analogs, derivatives, or salts
(e.g., mesylate salts) thereof inhibits microbial growth and
oxidation in the pharmaceutical composition, and it is believed to
bind to free drug in the composition. Deferoxamine also has been
shown to bind to phenolic compounds (see, e.g., Juven et al., J.
Appl. Bacteriol., 76(6), 626-31 (1994)). Paclitaxel, docetaxel,
propofol, and the like, are either phenolic like or have phenolic
or phenyl substituents. Therefore, it is believed that deferoxamine
can bind to or reduce the amount of free drug in the inventive
pharmaceutical composition, thereby also reducing or alleviating
irritation or pain upon injection.
[0023] The amount of deferoxamine, or its preferred salt, i.e., a
mesylate salt of deferoxamine, included in the composition will
depend on the active pharmaceutical agent and other excipients.
Desirably, the amount of deferoxamine, its salts, and analogs
thereof in the composition is an amount effective to inhibit
microbial growth and/or inhibit oxidation. As described above,
typically the pharmaceutical composition is prepared in liquid
form, and deferoxamine, it salts, and analogs thereof, is then
added in solution. Preferably, the pharmaceutical composition, in
liquid form, comprises from about 0.0001% to about 0.5% by weight
(e.g., about 0.005% by weight, about 0.1%, or about 0.25% by
weight) of deferoxamine, its salts, or its analogs. More
preferably, the composition, in liquid form, comprises like amounts
of the preferred deferoxamine salt, deferoxamine mesylate. Most
preferably, the pharmaceutical composition, in liquid form,
comprises about 0.1% by weight of deferoxamine mesylate. When the
composition is prepared in solid form, as described above, such as
by wet granulation, fluidized-bed drying, and other methods known
to those skilled in the art, deferoxamine mesylate preferably is
applied to the active pharmaceutical agent, and other excipients if
present, as a solution. The deferoxamine mesylate solution
preferably is from about 0.0001% to about 0.5% by weight (e.g.,
about 0.005% by weight, about 0.1%, or about 0.25% by weight) of
deferoxamine.
[0024] In keeping with the invention, the pharmaceutical
composition can include other agents, excipients, or stabilizers to
improve properties of the composition. For example, to increase
stability by increasing the negative zeta potential of
nanoparticles or nanodroplets, certain negatively charged
components may be added. Such negatively charged components
include, but are not limited to bile salts of bile acids consisting
of glycocholic acid, cholic acid, chenodeoxycholic acid,
taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic
acid, litocholic acid, ursodeoxycholic acid, dehydrocholic acid and
others; phospholipids including Lecithin (Egg yolk) based
phospholipids which include the following phosphatidylcholines:
palmitoyloleoylphosphatidylch- oline,
palmitoyllinoleoylphosphatidylcholine,
stearoyllinoleoylphosphatidy- lcholine
stearoyloleoylphosphatidylcholine, stearoylarachidoylphosphatidyl-
choline, and dipalmitoylphosphatidylcholine. Other phospholipids
including L-.alpha.-dimyristoylphosphatidylcholine (DMPC),
dioleoylphosphatidylchol- ine (DOPC), distearyolphosphatidylcholine
(DSPC), hydrogenated soy phosphatidylcholine (HSPC),
D-.alpha.-phosphatidylcholine, .beta.-acetyl-.gamma.-O-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.-acetyl-.gamma.-O-hexadecyl,
DL-.alpha.-phosphatidylcholine, .beta.-acetyl-.gamma.-O-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.-acetyl-65 -O-octadecyl,
L-.alpha.-phosphatidylcholine, .beta.-arachidonoyl-65 -O-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.-acetyl-65
-O-(octadec-9-cis-enyl), D-.alpha.-phosphatidylcholine,
.beta.-arachidonoyl-65 -O-palmitoyl, 3-sn-phosphatidylcholine,
2-arachidinoyl-1-stearoyl, L-.alpha.-phosphatidylcholine,
.beta.-arachidonoyl-65 -stearoyl, L-.alpha.-phosphatidylcholine,
diarachidoyl, L-.alpha.-phosphatidylcholine, dibehenoyl,
L-.alpha.-phosphatidylcholine,
.beta.-(cis-8,11,14-eicosatrienoyl)-65 -O-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.-oleoyl-65 -myristoyl,
L-.alpha.-phosphatidylcholine, .beta.-(pyren-1-yl)decanoyl-65
-palmitoyl, 3-sn-phosphatidyl-N,N-dimethylethanolamine,
1,2-dipalmitoyl, L-.alpha.-phosphatidylethanolamine,
diheptadecanoyl, 3-sn-phosphatidylethanolamine, 1,2-dilauroyl,
3-sn-phosphatidylethanolami- ne, 1,2-dimyristoyl,
3-sn-phosphatidylethanolamine, 1,2-dioleoyl,
3-sn-phosphatidylethanolamine, 1,2-dipalmitoyl,
L-.alpha.-phosphatidyleth- anolamine, dipalmitoyl,
L-.alpha.-phosphatidylethanolamine, dipalmitoyl, N-dansyl,
L-.alpha.-phosphatidylethanolamine, dipalmitoyl, N,N-dimethyl,
L-.alpha.-dimyristoylphosphatidylglycerol (sodium salt) (DMPG),
dipalmitoylphosphatidylglycerol (sodium salt) (DPPG),
distearoylphosphatidylglycerol (sodium salt) (DSPG),
N-(carbonyl-methoxypolyethylene glycol
2000)-1,2-distearoyl-sn-glycero-3-- phosphoethanolamine sodium
(MPEG-DSPE), L-.alpha.-phosphatidic acid, didecanoyl sodium salt,
L-.alpha.-phosphatidic acid, diheptadecanoyl sodium salt,
3-sn-phosphatidic acid, 1,2-dimyristoyl sodium salt,
L-.alpha.-phosphatidic acid, dioctanoyl sodium salt,
L-.alpha.-phosphatidic acid, dioleoyl sodium salt,
L-.alpha.-phosphatidic acid, dipalmitoyl sodium salt,
L-.alpha.-Phosphatidyl-DL-glycerol, dimyristoyl sodium salt,
L-.alpha.-phosphatidyl-DL-glycerol, dioleoyl sodium salt,
L-.alpha.-phosphatidyl-DL-glycerol, dipalmitoyl ammonium salt,
L-.alpha.-phosphatidyl-DL-glycerol, distearoyl ammonium salt,
L-.alpha.-phosphatidyl-DL-glycerol, .beta.-oleoyl-65 -palmitoyl
ammonium salt, L-.alpha.-phosphatidylinositol ammonium salt,
L-.alpha.-phosphatidylinositol sodium salt,
L-.alpha.-phosphatidyl-L-seri- ne, dioleoyl sodium salt,
L-.alpha.-phosphatidyl-L-serine, and dipalmitoyl sodium salt.
Negatively charged surfactants of emulsifiers are also suitable as
additives, e.g., sodium cholesteryl sulfate and the like.
[0025] The pharmaceutical agent (e.g., propofol) may be used alone
or dissolved in a water-immiscible solvent. A wide range of
water-immiscible solvents such as soybean, safflower, cottonseed,
corn, sunflower, arachis, castor, or olive oil may be used. The
preferred oil is a vegetable oil, wherein soybean oil is most
preferred. Soybean oil may be used in a range of 1% to 10% by
weight of the composition. Preferably soybean oil is present in the
pharmaceutical composition in an amount of about 3% by weight.
[0026] The inventive pharmaceutical composition can be stabilized
with a pharmaceutically acceptable surfactant. The term
"surfactants," as used herein, refers to surface active group(s) of
amphiphile molecules. Surfactants can be anionic, cationic,
nonionic, and zwitterionic. Any suitable surfactant can be included
in the inventive pharmaceutical composition. Suitable surfactants
include non-ionic surfactants such as phosphatides, polyoxyethylene
sorbitan esters, and tocopheryl polyethylene glycol succinate.
Preferable surfactants are egg lecithin, tween 80, and vitamin E-t
d-.alpha.-tocopheryl polyethylene glycol-1000 succinate (TPGS). For
soybean oil containing formulations, egg lecithin is preferred and
is no more than 1.2% by weight for a formulation containing 3%
soybean oil, preferably at 1.1% by weight of the composition. For
formulations without soybean oil, tween 80 or vitamin E-TPGS are
the preferred surfactants. Typically, 0.1 to 1.5% by weight of
tween 80 or 0.5 to 4% by weight of vitamin E-TPGS is suitable.
Preferably, 1.5% by weight of tween 80 or 1% by weight of vitamin
E-TPGS is used. Examples of other suitable surfactants are
described in, for example, Becher, Emulsions: Theory and Practice,
Robert E. Krieger Publishing, Malabar, Fla. (1965).
[0027] There are a wide variety of suitable formulations of the
inventive pharmaceutical composition (see, e.g., U.S. Pat. No.
5,916,596). The following formulations and methods are merely
exemplary and are in no way limiting. Formulations suitable for
oral administration can consist of (a) liquid solutions, such as an
effective amount of the compound dissolved in diluents, such as
water, saline, or orange juice, (b) capsules, sachets or tablets,
each containing a predetermined amount of the active ingredient, as
solids or granules, (c) suspensions in an appropriate liquid, and
(d) suitable emulsions. Tablet forms can include one or more of
lactose, mannitol, corn starch, potato starch, microcrystalline
cellulose, acacia, gelatin, colloidal silicon dioxide,
croscarmellose sodium, talc, magnesium stearate, stearic acid, and
other excipients, colorants, diluents, buffering agents, moistening
agents, preservatives, flavoring agents, and pharmacologically
compatible excipients. Lozenge forms can comprise the active
ingredient in a flavor, usually sucrose and acacia or tragacanth,
as well as pastilles comprising the active ingredient in an inert
base, such as gelatin and glycerin, or sucrose and acacia,
emulsions, gels, and the like containing, in addition to the active
ingredient, such excipients as are known in the art.
[0028] Formulations suitable for parenteral administration include
aqueous and non-aqueous, isotonic sterile injection solutions,
which can contain anti-oxidants, buffers, bacteriostats, and
solutes that render the formulation isotonic with the blood of the
intended recipient, and aqueous and non-aqueous sterile suspensions
that can include suspending agents, solubilizers, thickening
agents, stabilizers, and preservatives. The formulations can be
presented in unit-dose or multi-dose sealed containers, such as
ampules and vials, and can be stored in a freeze-dried
(lyophilized) condition requiring only the addition of the sterile
liquid excipient, for example, water, for injections, immediately
prior to use. Extemporaneous injection solutions and suspensions
can be prepared from sterile powders, granules, and tablets of the
kind previously described. Injectable formulations are
preferred.
[0029] Formulations suitable for aerosol administration comprise
the inventive pharmaceutical composition include aqueous and
non-aqueous, isotonic sterile solutions, which can contain
anti-oxidants, buffers, bacteriostats, and solutes, as well as
aqueous and non-aqueous sterile suspensions that can include
suspending agents, solubilizers, thickening agents, stabilizers,
and preservatives, alone or in combination with other suitable
components, which can be made into aerosol formulations to be
administered via inhalation. These aerosol formulations can be
placed into pressurized acceptable propellants, such as
dichlorodifluoromethane, propane, nitrogen, and the like. They also
can be formulated as pharmaceuticals for non-pressured
preparations, such as in a nebulizer or an atomizer.
[0030] Other suitable formulations are possible, for example,
suppositories can be prepared by use of a variety of bases such as
emulsifying bases or water-soluble bases. Formulations suitable for
vaginal administration can be presented as pessaries, tampons,
creams, gels, pastes, foams, or spray formulas containing, in
addition to the active ingredient, such carriers as are known in
the art to be appropriate.
[0031] In a preferred embodiment of the invention, the
pharmaceutical composition is formulated to have a pH range of 4.5
to 9.0, and more preferably a pH of 5.0 to 8.0. The pharmaceutical
composition can also be made to be isotonic with blood by the
addition of a suitable tonicity modifier, such as glycerol.
Moreover, the pharmaceutically acceptable carrier preferably also
comprises pyrogen-free water or water for injection, USP.
Preferably, the inventive pharmaceutical composition is prepared as
a sterile aqueous formulation, a nanoparticle, an oil-in-water
emulsion, or a water-in-oil emulsion. Most preferably, the
pharmaceutical composition is an oil-in-water emulsion.
[0032] For a pharmaceutical composition comprising propofol, in
accordance with the invention, an oil-in-water emulsion is prepared
by dissolving propofol in a water-immiscible solvent alone, and
preparing an aqueous phase containing albumin, deferoxamine, a
surfactant, and other water-soluble ingredients, and mixing the oil
with the aqueous phase. The crude emulsion is high pressure
homogenized at pressures of 10,000 to 25,000 psi and recirculated
for 5 to 20 cycles to form an ideal emulsion. The preferred
pressure is 15,000 to 20,000 psi., and more preferably 10,000 psi.
The crude emulsion may be recirculated from 7 to 15 cycles and is
preferably recirculated at 15 cycles. Alternatively, discrete
passes through a homogenizer may be used.
[0033] Preferably, the inventive pharmaceutical composition can
have a particle or droplet size less than about 200 nanometers
(nm). For example, in the case of paclitaxel, docetaxel, rapamycin,
cyclosporine, propofol and others, the mean size of these
dispersions is less than 200 nm.
[0034] The invention further provides a method for reducing one or
more side effects associated with administration of a
pharmaceutical composition to a human. The method comprises
administering to a human a pharmaceutical composition comprising a
pharmaceutical agent and a pharmaceutically acceptable carrier,
wherein the pharmaceutically acceptable carrier comprises albumin
and deferoxamine. Descriptions of the pharmaceutical composition,
pharmaceutical agent, and pharmaceutically acceptable carrier, and
components thereof set forth above in connection with the inventive
pharmaceutical composition, also are applicable to those same
aspects of the inventive method.
[0035] The dose of the inventive pharmaceutical composition
administered to a human, in the context of the invention, will vary
with the particular pharmaceutical composition, the method of
administration, and the particular site being treated. The dose
should be sufficient to effect a desirable response, such as a
therapeutic or prophylactic response against a particular disease,
or, when the pharmaceutical agent is an anaesthesia, such as
propofol, an anesthetic response, within a desirable time
frame.
[0036] While any suitable means of administering the pharmaceutical
composition to the human can be used within the context of the
invention, preferably the inventive pharmaceutical composition is
administered to the human via intravenous administration,
intra-arterial administration, intrapulmonary administration, oral
administration, inhalation, intravesicular administration,
intramuscular administration, intra-tracheal administration,
subcutaneous administration, intraocular administration,
intrathecal administration, or transdermal administration. For
example, the inventive pharmaceutical composition can be
administered by inhalation to treat conditions of the respiratory
tract. There are minimal side-effects associated with the
inhalation of the inventive pharmaceutical composition, as albumin
is a natural component in the lining and secretions of the
respiratory tract. The inventive composition can be used to treat
respiratory conditions such as pulmonary fibrosis, broncheolitis
obliterans, lung cancer, bronchoalveolar carcinoma, and the
like.
[0037] The inventive method results in the reduction of one or more
side effects associated with administration of a pharmaceutical
composition to a human. Such side effects include, for example,
myelosuppression, neurotoxicity, hypersensitivity, inflammation,
venous irritation, phlebitis, pain, skin irritation, and
combinations thereof. These side effects, however, are merely
exemplary, and other side effects, or combination of side effects,
associated with various pharmaceutical agents can be reduced or
avoided by the use of the novel compositions and methods of the
present invention.
[0038] The invention further provides a method for inhibiting
microbial growth in a pharmaceutical composition. By "inhibiting
microbial growth" is meant either a complete elimination of
microbes from the pharmaceutical composition, or a reduction in the
amount or rate of microbial growth in the pharmaceutical
composition. The method comprises preparing a pharmaceutical
composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises deferoxamine, its salts, its analogs,
and combinations thereof, in an amount effective for inhibiting
microbial growth in the pharmaceutical composition. In addition,
the invention provides a method for inhibiting oxidation of a
pharmaceutical composition. This method comprises preparing a
pharmaceutical composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises deferoxamine, its salts, its analogs,
and combinations thereof, in an amount effective for inhibiting
oxidation of the pharmaceutical composition. Descriptions of the
pharmaceutical composition, pharmaceutical agent, and
pharmaceutically acceptable carrier, and components thereof set
forth above in connection with the inventive pharmaceutical
composition, also are applicable to those same aspects of the
inventive method.
[0039] The amount of deferoxamine, or its preferred salt, a
mesylate salt of deferoxamine, included in the composition will
depend on the active pharmaceutical agent and other excipients.
Desirably, the amount of deferoxamine, its salts, and analogs
thereof in the composition is an amount effective to inhibit
microbial growth and/or inhibit oxidation. As described above,
typically, the pharmaceutical composition is prepared in liquid
form, and deferoxamine, it salts, and analogs thereof, is then
added in solution. Preferably, the pharmaceutical composition, in
liquid form, comprises from about 0.0001% to about 0.5% by weight
(e.g., about 0.005% by weight, about 0.1%, or about 0.25% by
weight) of deferoxamine, its salts, or its analogs. More
preferably, the composition, in liquid form, comprises like amounts
of the preferred deferoxamine salt, deferoxamine mesylate. Most
preferably, the pharmaceutical composition, in liquid form,
comprises about 0.5% by weight of deferoxamine mesylate. When the
composition is prepared in solid form, as described above, such as
by wet granulation, fluidized-bed drying, and other methods known
to those skilled in the art, deferoxamine mesylate preferably is
applied to the active pharmaceutical agent, and other excipients if
present, as a solution. The deferoxamine mesylate solution
preferably is from about 0.0001% to about 0.5% by weight (e.g.,
about 0.005% by weight, about 0.1%, or about 0.25% by weight) of
deferoxamine.
[0040] The invention also provides a method for enhancing transport
of a pharmaceutical agent to the site of an infirmity, which method
comprises administering to a human a pharmaceutical composition
comprising a pharmaceutical agent and a pharmaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
albumin, and wherein the ratio of albumin to pharmaceutical agent
in the pharmaceutical composition is about 18:1 or less. The
invention further provides a method for enhancing binding of a
pharmaceutical agent to a cell in vitro or in vivo, which method
comprises administering to said cell in vitro or in vivo a
pharmaceutical composition comprising a pharmaceutical agent and a
pharmaceutically acceptable carrier, wherein the pharmaceutically
acceptable carrier comprises albumin, and wherein the ratio of
albumin to pharmaceutical agent in the pharmaceutical composition
is about 18:1 or less. Descriptions of the pharmaceutical
composition, pharmaceutical agent, pharmaceutically acceptable
carrier, administration routes, and components thereof set forth
above in connection with the inventive pharmaceutical composition
and inventive method also are applicable to those same aspects of
the transport and binding methods.
[0041] In the methods for enhancing transport of a pharmaceutical
agent to the site of an infirmity or for enhancing binding of a
pharmaceutical agent to a cell, the pharmaceutically acceptable
carrier preferably comprises albumin, most preferably human serum
albumin. Not to adhere to any one particular theory, it is believed
that the ratio of protein, e.g., human serum albumin, to
pharmaceutical agent in the pharmaceutical composition affects the
ability of the pharmaceutical agent to bind and transport the
pharmaceutical agent to a cell. In this regard, higher ratios of
protein to pharmaceutical agent generally are associated with poor
cell binding and transport of the pharmaceutical agent, which
possibly is the result of competition for receptors at the cell
surface. The ratio of protein, e.g., albumin, to active
pharmaceutical agent must be such that a sufficient amount of
pharmaceutical agent binds to, or is transported by, the cell.
Exemplary ranges for protein-drug preparations are protein to drug
ratios (w/w) of 0.01:1 to about 100:1. More preferably, the ratios
are in the range of 0.02:1 to about 40:1. While the ratio of
protein to pharmaceutical agent will have to be optimized for
different protein and pharmaceutical agent combinations, generally
the ratio of protein, e.g., albumin, to pharmaceutical agent is
about 18:1 or less (e.g., about 15:1, about 10:1, about 5:1, or
about 3:1). More preferably, the ratio is about 0.2:1 to about
12:1. Most preferably, the ratio is about 1:1 to about 9:1.
Preferably, the formulation is essentially free of cremophor, and
more preferably free of Cremophor EL.RTM. (BASF). Cremophor EL.RTM.
is a non-ionic emulsifying agent that is a polyether of castor oil
and ethylene oxide. As described above, cremophor typically is used
as a solvent for paclitaxel, and is associated with side effects
that can be severe (see, e.g., Gelderblom et al., supra).
[0042] The pharmaceutical agent can be any suitable pharmaceutical
agent described herein (e.g., propofol, paclitaxel, or docetaxel).
In addition, the pharmaceutical agent can be a nucleic acid
sequence, most preferably a DNA sequence. In this regard, the
inventive pharmaceutical composition can be used to transport genes
to a cell by way of a receptor mediated/caveolar/vescicular
transport. In order to transport DNA sequences, such as genes or
other genetic material, including but not limited to plasmids or
c-DNA, into a cell (e.g. an endothelial cell or a tumor cell),
pharmaceutical compositions comprising albumin in combination with
genetic material can be prepared. Since tumor cells and other cells
in sites of inflammation have high uptake for proteins, the genetic
material is preferentially taken up into these cell types and may
be incorporated into the genetic material of the cell for a useful
therapeutic effect. The use of proteins, such as human serum
albumin, serves as a non-viral vector for the delivery of genetic
material without the risk of virus-associated diseases or side
effects. For example, a pharmaceutical composition comprising the
nucleic acid sequence encoding .beta.-galactosidase or green
fluorescent protein (GFP) and albumin can be prepared and contacted
with endothelial cells derived from human umbilical vein or human
lung microvessels to facilitate incorporation of the nucleic acid
sequence into the endothelial cells. Incorporation of the nucleic
acid sequence can be detected using methods known in the art, such
as, for example, fluorescence or staining.
[0043] In the inventive method for enhancing transport of a
pharmaceutical agent to the site of an infirmity, the infirmity can
be any suitable disease or condition. Preferably, the infirmity is
cancer, cardiovascular disease, or arthritis.
[0044] In the inventive method for enhancing binding of a
pharmaceutical agent to a cell in vitro or in vivo, the
pharmaceutical composition is administered to a cell in vitro or in
vivo. Desirably, the cell is an animal cell. More preferably the
cell is a mammalian cell, and most preferably the cell is a human
cell. The pharmaceutical composition preferably is administered to
a cell in vivo. The cell can be any suitable cell that is a
desirable target for administration of the pharmaceutical
composition. For example, the cell can be located in or derived
from tissues of the digestive system including, for example, the
esophagus, stomach, small intestine, colon, rectum, anus, liver,
gall bladder, and pancreas. The cell also can be located in or
derived from tissues of the respiratory system, including, for
example, the larynx, lung, and bronchus. The cell can be located in
or derived from, for example, the uterine cervix, the uterine
corpus, the ovary vulva, the vagina, the prostate, the testis, and
the penis, which make up the male and female genital systems, and
the urinary bladder, kidney, renal pelvis, and ureter, which
comprise the urinary system. The cell can be located in or derived
from tissues of the cardiovascular system, including, for example,
endothelial cells and cardiac muscle cells. The cell also can be
located in or derived from tissues of the lymphoid system (e.g.,
lymph cells), the nervous system (e.g., neurons or glial cells),
and the endocrine system (e.g., thyroid cells). Preferably, the
cell is located in or derived from tissues of the cardiovascular
system. Most preferably, the cell is an endothelial cell. In the
context of the inventive method for enhancing transport and
enhancing binding of a pharmaceutical agent to a cell, the
pharmaceutical composition desirably contacts more than one
cell.
[0045] In another aspect of the invention, the inventive methods
for enhancing transport and enhancing binding of a pharmaceutical
agent to a cell can be used to treat tumor cells. Tumor cells
exhibit an enhanced uptake of proteins including, for example,
albumin and transferrin, as compared to normal cells. Since tumor
cells are dividing at a rapid rate, they require additional
nutrient sources compared to normal cells. Tumor studies of the
inventive pharmaceutical compositions containing paclitaxel and
human serum albumin showed high uptake of albumin-paclitaxel into
tumors. This has been found to be due to the previously
unrecognized phenomenon of the albumin-drug transport by
glycoprotein 60 ("gp60") receptors, which are specific for
albumin.
[0046] Thus, in accordance with another aspect of the present
invention, the albumin-specific gp60 receptor and other protein
transport receptors that are present on tumor cells can be used as
a target to inhibit tumor growth. By blocking the gp60 receptor
using antibodies against the gp60 receptor or other large or small
molecule compounds that bind, block, or inactivate gp60 and other
protein transport receptors on tumor cells or tumor endothelial
cells, it is possible to block the transport of proteins to these
cells and thereby reduce their growth rate and cause cell death.
Blocking of this mechanism thus results in the treatment of a
subject (e.g., a human) with cancer or another disease.
Identification of blocking/binding of the specific protein receptor
is done by screening any number of compounds against the isolated
gp60 or other receptors, such as gp16 orgp30, or by using a whole
cell preparation. In addition, suitable animal models also can be
used for this purpose, such as, for example, mice containing
"knock-out" mutations of the genes encoding gp60 or caveolin-1, or
other proteins that are specific for transport. Thus, method of
identification of compounds that block or bind gp60, gp16, gp30, or
other protein receptors are within the scope of the invention.
[0047] In addition, compounds that block or bind the gp60 receptor
or other protein receptors can be used in the treatment of several
diseases, including cancer. With respect to the treatment of
cancer, the blocking or binding compound may be used as a single
agent or in combination with other standard chemotherapy or
chemotherapies. For example, it is useful to treat the cancer with
conventional chemotherapy, or with the inventive albumin-drug
pharmaceutical compositions (which show high accumulation in
tumors), followed by compounds that block the transport of proteins
to the tumor cell. Blocking compounds can be administered prior to,
or in conjunction with, other chemotherapeutic or anticancer
agents. Thus, any compounds that can block or bind the gp60
receptor, or other protein receptors, are within the scope of the
present invention.
[0048] The inventive albumin-drug compositions, such as e.g.,
albumin-paclitaxel, albumin-docetaxel, albumin-epothilone,
albumin-camptothecin, or albumin-rapamycin, and others, are useful
in the treatment of diseases. It is believed that such drug
compositions are effective due to increased receptor mediated
transport of the protein-drug composition to the required site, for
example a tumor. Without wishing to be bound to any particular
theory, the transport of a protein-drug composition by receptor
mediated transport resulting in a therapeutic effect is believed to
be the mechanism for transport of for example, albumin-paclitaxel
compositions to a tumor, as well as albumin-paclitaxel and
albumin-rapamycin transport across the lung. Transport is effected
by the presence of gp60, gp16, or gp30 in such tissues.
Accordingly, drugs and protein-drug compositions whose transport to
sites of disease, e.g., inflammation (e.g., arthritis) or tumors is
associated with gp60, gp16, or gp30 receptors and that result in a
therapeutic effect are contemplated as compositions of the present
invention.
[0049] In accordance with another aspect of the present invention,
endothelial cells can be co-cultured with cells having a specific
function. Incubation of endothelial cells with other cell types
such as islet cells, hepatocytes, neuroendocrine cells, and others
allows for required transport of components such as proteins and
other beneficial components to these cells. The endothelial cells
provide for transport of these components to the cultured cell
types in order to simulate in vivo conditions, i.e., where these
cell types would normally be in close proximity to endothelial
cells and would depend on the endothelial cells for transport of
nutrients, growth factors, hormone signals, etc. that are required
for their proper function. It has previously not been possible to
adequately culture these different cell types and obtain
physiological performance when endothelial cells were absent. The
presence of endothelial cells in culture with desired cell types
allows for differentiation and proper functioning of islets,
hepatocytes, or neuroendocrine tissue in vitro or ex vivo. Thus it
is found that coculture of endothelial cells with islets results in
islets with improved physiological properties e.g., ability to
secrete insulin, when compared with those cultured in the absence
of endothelial cells. This tissue can then be used ex vivo or
transplanted in vivo to treat diseases caused by lack of adequate
cellular function (e.g., diabetes in the case of islet cells,
hepatic dysfunction in the case of hepatocytes, and neuroendocrine
disorders or pain relief in the case of neuroendocrine cells).
Cells originating from other tissues and organs (as described
above) may also be cocultured with endothelial cells to provide the
same benefit. In addition, the coculture may be utilized to
incorporate genetic material into the target cell types. The
presence of albumin in these cultures is found to be greatly
beneficial.
[0050] The following examples further illustrate the invention but,
of course, should not be construed as in any way limiting its
scope.
EXAMPLE 1
[0051] This example demonstrates the preparation of pharmaceutical
compositions comprising paclitaxel and albumin. Preparation of
paclitaxel-albumin compositions is described in U.S. Pat. Nos.
5,439,686 and 5,916,596, which are incorporated in their entirety
by reference. Specifically, 30 mg of paclitaxel was dissolved in
3.0 ml methylene chloride. The solution was added to 27.0 ml of
human serum albumin solution (2% w/v). Deferoxamine was added as
necessary. The mixture was homogenized for 5 minutes at low RPM
(Vitris homogenizer, model Tempest I.Q.) in order to form a crude
emulsion, and then transferred into a high pressure homogenizer
(Avestin). The emulsification was performed at 9000-40,000 psi
while recycling the emulsion for at least 5 cycles. The resulting
system was transferred into a rotary evaporator, and methylene
chloride was rapidly removed at 40.degree. C., at reduced pressure
(30 mm Hg) for 20-30 minutes. The resulting dispersion was
translucent, and the typical average diameter of the resulting
paclitaxel particles was in the range 50-220 nm (Z-average, Malvern
Zetasizer). The dispersion was further lyophilized for 48 hrs. The
resulting cake could be easily reconstituted to the original
dispersion by addition of sterile water or saline. The particle
size after reconstitution was the same as before
lyophilization.
[0052] It should be recognized that the amounts, types and
proportions of drug, solvents, proteins used in this example are
not limiting in any way. When compared to toxicity of paclitaxel
dissolved in cremophor formulations, the inventive pharmaceutical
composition containing albumin showed substantially lower
toxicity.
EXAMPLE 2
[0053] This example demonstrates the preparation of a
pharmaceutical composition comprising amiodarone and albumin. 30 mg
of amiodarone was dissolved in 3.0 ml methylene chloride. The
solution was added to 27.0 ml of human serum albumin solution (1%
w/v). Deferoxamine was added as necessary. The mixture was
homogenized for 5 minutes at low RPM (Vitris homogenizer, model
Tempest I.Q.) in order to form a crude emulsion, and then
transferred into a high pressure homogenizer (Avestin). The
emulsification was performed at 9000-40,000 psi while recycling the
emulsion for at least 5 cycles. The resulting system was
transferred into a rotary evaporator, and methylene chloride was
rapidly removed at 40.degree. C., at reduced pressure (30 mm Hg)
for 20-30 minutes. The resulting dispersion was translucent, and
the typical average diameter of the resulting amiodarone particles
was in the range 50-220 nm (Z-average, Malvern Zetasizer). The
dispersion was further lyophilized for 48 hrs. The resulting cake
was easily reconstituted to the original dispersion by addition of
sterile water or saline. The particle size after reconstitution was
the same as before lyophilization.
[0054] It should be recognized that the amounts, types and
proportions of drug, solvents, proteins used in this example are
not limiting in anyway. When compared to toxicity of amiodarone
dissolved in tween formulations, the inventive pharmaceutical
composition with albumin showed substantially lower toxicity.
EXAMPLE 3
[0055] This example demonstrates the preparation of pharmaceutical
compositions comprising liothyronine and albumin compositions.
Liothyronine (or suitable salt) was dissolved in an aqueous
alcoholic solution or alkaline solution at a concentration of
0.5-50 mg/ml. The alcoholic (or alkaline) solution was added to an
albumin solution (0.1-25% w/v) and agitated. Agitation was low
shear with a stirrer or high shear using a sonicator or a
homogenizer. At low concentrations of liothyronine, (5-1000
.mu.g/ml) clear solutions were obtained. As the concentration was
increased, a milky stable suspension was obtained. These solutions
or suspensions were filtered through a sterilizing filter. Organic
solvents were removed by evaporation or other suitable method.
EXAMPLE 4
[0056] This example demonstrates the preparation of pharmaceutical
compositions comprising rapamycin and albumin. 30 mg of rapamycin
was dissolved in 2 ml chloroform/ethanol. The solution was then
added into 27.0 ml of a human serum albumin solution (3% w/v). The
mixture was homogenized for 5 minutes at low RPM (Vitris
homogenizer model Tempest I.Q.) in order to form a crude emulsion,
and then transferred into a high pressure homogenizer (Avestin).
The emulsification was performed at 9000-40,000 psi while recycling
the emulsion for at least 5 cycles. The resulting system was
transferred into a Rotavap and solvent was rapidly removed at
40.degree. C., at reduced pressure (30 mm Hg) for 20-30 minutes.
The resulting dispersion was translucent and the typical average
diameter of the resulting particles was in the range 50-220 nm
(Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, proteins used in
this example are not limiting in anyway.
EXAMPLE 5
[0057] This example demonstrates the preparation of a
pharmaceutical composition comprising epothilone B and albumin. 30
mg of epothilone B was dissolved in 2 ml chloroform/ethanol. The
solution was then added into 27.0 ml of a human serum albumin
solution (3% w/v). Deferoxamine was added as necessary. The mixture
was homogenized for 5 minutes at low RPM (Vitris homogenizer model
Tempest I.Q.) in order to form a crude emulsion, and then
transferred into a high pressure homogenizer (Avestin). The
emulsification was performed at 9000-40,000 psi while recycling the
emulsion for at least 5 cycles. The resulting system was
transferred into a Rotavap and solvent was rapidly removed at
40.degree. C., at reduced pressure (30 mm Hg) for 20-30 minutes.
The resulting dispersion was translucent and the typical average
diameter of the resulting particles was in the range 50-220 nm
(Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, proteins used in
this example are not limiting. When compared to toxicity of
epothilone B dissolved in cremophor formulations, the
pharmaceutical composition comprising albumin showed substantially
lower toxicity.
EXAMPLE 6
[0058] This example demonstrates the preparation of pharmaceutical
compositions comprising colchicine dimer and albumin. 30 mg of
colchicine-dimer was dissolved in 2 ml chloroform/ethanol. The
solution was then added into 27.0 ml of human serum albumin
solution (3% w/v). Deferoxamine was added as necessary. The mixture
was homogenized for 5 minutes at low RPM (Vitris homogenizer model
Tempest I.Q.) in order to form a crude emulsion, and then
transferred into a high pressure homogenizer (Avestin). The
emulsification was performed at 9000-40,000 psi while recycling the
emulsion for at least 5 cycles. The resulting system was
transferred into a Rotavap and solvent was rapidly removed at
40.degree. C., at reduced pressure (30 mm Hg) for 20-30 minutes.
The resulting dispersion was translucent and the typical average
diameter of the resulting particles was in the range 50-220 nm
(Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, proteins used in
this example are not limiting. When compared to toxicity of the
colchicines dimer dissolved in tween, the pharmaceutical
composition comprising albumin showed substantially lower
toxicity.
EXAMPLE 7
[0059] This example demonstrates the preparation of pharmaceutical
compositions comprising docetaxel and albumin. 30 mg of docetaxel
was dissolved in 2 ml chloroform/ethanol. The solution was then
added into 27.0 ml of human serum albumin solution (3% w/v).
Deferoxamine was added as necessary. The mixture was homogenized
for 5 minutes at low RPM (Vitris homogenizer model Tempest I.Q.) in
order to form a crude emulsion, and then transferred into a high
pressure homogenizer (Avestin). The emulsification was performed at
9000-40,000 psi while recycling the emulsion for at least 5 cycles.
The resulting system was transferred into a Rotavap and solvent was
rapidly removed at 40.degree. C., at reduced pressure (30 mm Hg)
for 20-30 minutes. The resulting dispersion was translucent and the
typical average diameter of the resulting particles was in the
range 50-220 run (Z-average, Malvern Zetasizer). The dispersion was
further lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, and proteins used
in this example are not limiting. When compared to toxicity of the
docetaxel dissolved in tween/ethanol which is the standard solvent
for this drug, the pharmaceutical composition comprising albumin
showed substantially lower toxicity.
EXAMPLE 8
[0060] This example demonstrates the preparation of pharmaceutical
compositions comprising docetaxel and albumin. 150 mg of docetaxel
was dissolved in 1 ml ethyl acetate/butyl acetate and 0.5 ml of an
oil for example soybean oil or vitamin E oil. Other ratios of
solvents and oils were used and these compositions are also
contemplated as part of the invention. A small quantity of a
negatively charged component was also optionally added, e.g.,
benzoic acid (0.001%-0.5%) The solution was then added into 27.0 ml
of human serum albumin solution (5% w/v). Deferoxamine was added as
necessary. The mixture was homogenized for 5 minutes at low RPM
(Vitris homogenizer model Tempest I.Q.) in order to form a crude
emulsion, and then transferred into a high pressure homogenizer
(Avestin). The emulsification was performed at 9000-40,000 psi
while recycling the emulsion for at least 5 cycles. The resulting
system was transferred into a Rotavap and solvent was rapidly
removed at 40.degree. C., at reduced pressure (30 mm Hg) for 20-30
minutes. The resulting dispersion was translucent and the typical
average diameter of the resulting particles was in the range 50-220
nm (Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, proteins used in
this example are not limiting. When compared to toxicity of the
docetaxel dissolved in tween/ethanol which is the standard solvent
for this drug, the pharmaceutical composition comprising albumin
showed substantially lower toxicity.
EXAMPLE 9
[0061] This example demonstrates the preparation of pharmaceutical
compositions comprising a taxane IDN5390 and albumin. 150 mg of
IDN5390 was dissolved in 1 ml ethyl acetatelbutyl acetate and 0.5
ml of an oil for example soybean oil or vitamin E oil. Other ratios
of solvents and oils were used and these compositions are also
contemplated as part of the invention. A small quantity of a
negatively charged component was also optionally added, e.g.,
benzoic acid (0.001%-0.5%) The solution was then added into 27.0 ml
of human serum albumin solution (5% w/v). Deferoxamine was added as
necessary. The mixture was homogenized for 5 minutes at low RPM
(Vitris homogenizer model Tempest I.Q.) in order to form a crude
emulsion, and then transferred into a high pressure homogenizer
(Avestin). The emulsification was performed at 9000-40,000 psi
while recycling the emulsion for at least 5 cycles. The resulting
system was transferred into a Rotavap and solvent was rapidly
removed at 40.degree. C., at reduced pressure (30 mm Hg) for 20-30
minutes. The resulting dispersion was translucent and the typical
average diameter of the resulting particles was in the range 50-220
nm (Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, proteins used in
this example are not limiting. When compared to toxicity of the
IDN5390 dissolved in tween, the pharmaceutical composition
comprising albumin showed substantially lower toxicity.
EXAMPLE 10
[0062] This example demonstrates the preparation of pharmaceutical
compositions comprising a taxane IDN5109 and albumin. 150 mg of
IDN5109 was dissolved in 2 ml chloroform/ethanol. Other ratios of
solvents and oils were used and these compositions are also
contemplated as part of the invention. A small quantity of a
negatively charged component was also optionally added, e.g.,
benzoic acid (0.001%-0.5%) The solution was then added into 27.0 ml
of human serum albumin solution (5% w/v). Deferoxamine was added as
necessary. The mixture is homogenized for 5 minutes at low RPM
(Vitris homogenizer model Tempest I.Q.) in order to form a crude
emulsion, and then transferred into a high pressure homogenizer
(Avestin). The emulsification was performed at 9000-40,000 psi
while recycling the emulsion for at least 5 cycles. The resulting
system was transferred into a Rotavap and solvent was rapidly
removed at 40.degree. C., at reduced pressure (30 mm Hg) for 20-30
minutes. The resulting dispersion was translucent and the typical
average diameter of the resulting particles was in the range 50-220
nm (Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, and proteins used
in this example are not limiting. When compared to toxicity of the
IDN5109 dissolved in tween, the pharmaceutical composition
comprising albumin showed substantially lower toxicity.
EXAMPLE 11
[0063] This example demonstrates the preparation of a
pharmaceutical composition comprising 10-hydroxy camptothecin
(10HC) and albumin. 30 mg of 10-HC was dissolved in 2.0 ml
DMF/methylene chloride/soybean oil. The solution was then added
into 27.0 ml of a human serum albumin solution (3% w/v). The
mixture was homogenized for 5 minutes at low RPM (Vitris
homogenizer model Tempest I.Q.) in order to form a crude emulsion,
and then transferred into a high pressure homogenizer (Avestin).
The emulsification was performed at 9000-40,000 psi while recycling
the emulsion for at least 5 cycles. The resulting system was
transferred into a Rotavap and solvent was rapidly removed at
40.degree. C., at reduced pressure (30 mm Hg) for 20-30 minutes.
The resulting dispersion was translucent and the typical average
diameter of the resulting particles was in the range 50-220 nm
(Z-average, Malvern Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. The particle size after reconstitution was the
same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, proteins used in
this example are not limiting in anyway.
EXAMPLE 12
[0064] This example demonstrates the preparation of a
pharmaceutical composition comprising cyclosporine and albumin. 30
mg of cyclosporine was dissolved in 3.0 ml methylene chloride. The
solution was then added into 27.0 ml of a human serum albumin
solution (1% w/v). The mixture was homogenized for 5 minutes at low
RPM (Vitris homogenizer model Tempest I.Q.) in order to form a
crude emulsion, and then transferred into a high pressure
homogenizer (Avestin). The emulsification was performed at
9000-40,000 psi while recycling the emulsion for at least 5 cycles.
The resulting system was transferred into a Rotavap and methylene
chloride was rapidly removed at 40.degree. C., at reduced pressure
(30 mm Hg) for 20-30 minutes. The resulting dispersion was
translucent and the typical average diameter of the resulting
particles was in the range 50-220 nm (Z-average, Malvern
Zetasizer). The dispersion was further lyophilized for 48 hours.
The resulting cake was easily reconstituted to the original
dispersion by addition of sterile water or saline. The particle
size after reconstitution was the same as before
lyophilization.
EXAMPLE 13
[0065] This example demonstrates the preparation of a
pharmaceutical composition containing oil and comprising
cyclosporine and albumin. 30 mg of cyclosporine was dissolved in
3.0 ml of a suitable oil (sesame oil containing 10% orange oil).
The solution was then added into 27.0 ml of a human serum albumin
solution (1% v/w). The mixture was homogenized for 5 minutes at low
RPM (Vitris homogenizer, model Tempest I.Q.) in order to form a
crude emulsion, and then transferred into a high pressure
homogenizer (Avestin). The emulsification as performed at
9000-40,000 psi while recycling the emulsion for at least 5 cycles.
The resulting dispersion had a typical average diameter in range of
50-220 nm (Z-average, Malvern Zetasizer). The dispersion was used
directly or lyophilized for 48 hours by optionally adding a
suitable cryoprotectant. The resulting cake was easily
reconstituted to the original dispersion by addition of sterile
water or saline. It should be recognized that the amounts, types
and proportions of drug, solvents, and proteins used in this
example are not limiting in anyway.
EXAMPLE 14
[0066] This example demonstrates the preparation of a
pharmaceutical composition comprising amphotericin and albumin. 30
mg of amphotericin was dissolved in 3.0 ml methyl
pyrrolidinone/methylene chloride. The solution was added to 27.0 ml
of a human serum albumin solution (1% w/v). The mixture was
homogenized for 5 minutes at low RPM (Vitris homogenizer, model
Tempest I.Q.) in order to form a crude emulsion, and then
transferred into a high pressure homogenizer (Avestin). The
emulsification was performed at 9000-40,000 psi while recycling the
emulsion for at least 5 cycles. The resulting system was
transferred into a rotary evaporator, and solvent was rapidly
removed at 40.degree. C., at reduced pressure (30 mm Hg) for 20-30
minutes. The resulting dispersion was translucent, and the typical
average diameter of the resulting amphotericin particles was
between 50-220 nm (Z-average, Malvern Zetasizer). The dispersion
was further lyophilized for 48 hrs. The resulting cake could be
easily reconstituted to the original dispersion by addition of
sterile water or saline. The particle size after reconstitution was
the same as before lyophilization. It should be recognized that the
amounts, types and proportions of drug, solvents, and proteins used
in this example are not limiting in anyway. Addition of other
components such as lipids, bile salts, etc., also resulted in
suitable formulations.
EXAMPLE 15
[0067] This example demonstrates preclinical pharmacokinetics and
pharmacodynamics of a pharmaceutical composition comprising albumin
and paclitaxel.
[0068] Several preclinical pharmacokinetic studies in mice and rats
were conducted to evaluate the possible advantages of
albumin-paclitaxel pharmaceutical compositions over
cremophor-paclitaxel (Taxol) pharmaceutical compositions. These
studies demonstrated: (1) that the pharmacokinetics of
albumin-paclitaxel in rats was linear, whereas Taxol
pharmacokinetics were non-linear with respect to dose, (2)
pharmaceutical compositions comprising albumin and paclitaxel
exhibited a lower plasma AUC and C.sub.max, suggesting more rapid
distribution of albumin-paclitaxel compositions to tissues compared
with Taxol (excretion is similar), (3) pharmaceutical compositions
comprising albumin and paclitaxel exhibited a lower C.sub.max,
which possibly accounts for the reduced toxicities associated with
peak blood levels relative to Taxol, (4) the half-life of
pharmaceutical compositions comprising albumin and paclitaxel
exhibited was approximately 2-fold higher in rats and 4-fold higher
in tumor bearing mice relative to Taxol, and (5) the metabolism of
paclitaxel in pharmaceutical compositions comprising albumin and
paclitaxel was slower than in Taxol pharmaceutical compositions. At
24 hours post-injection in rats, 44% of total radioactivity was
still associated with paclitaxel for pharmaceutical compositions
comprising albumin and paclitaxel, compared to only 22% for Taxol.
The ultimate effect of the above pharmacodynamics, i.e., enhanced
intra-cellular uptake, prolonged half-life and slower metabolism
for pharmaceutical compositions comprising albumin and paclitaxel
exhibited resulted in a tumor AUC 1.7-fold higher, tumor C.sub.max
1.2-fold higher, and tumor half-life 1.7-fold longer than for Taxol
in tumor bearing mice.
EXAMPLE 16
[0069] This example demonstrates reduced side effects and reduced
toxicity associated with pharmaceutical compositions comprising
paclitaxel and albumin.
[0070] Due to the unique nature of pharmaceutical compositions
comprising paclitaxel and albumin in the absence of cremophor, the
toxicity of pharmaceutical compositions comprising paclitaxel and
albumin is substantially lower than Taxol. In preclinical studies
in mice and rats, a single dose acute toxicity study in mice showed
an LD.sub.50 dose approximately 59 times greater for pharmaceutical
compositions comprising paclitaxel and albumin than for Taxol. In a
multiple dose toxicity study in mice, the LD.sub.50 dose was
approximately 10-fold greater for pharmaceutical compositions
comprising paclitaxel and albumin than for Taxol. A further study
evaluated the degree of myelosuppression in rats treated with
pharmaceutical compositions comprising paclitaxel and albumin and
Taxol. The results showed that at equi-dose, pharmaceutical
compositions comprising paclitaxel and albumin produced
considerably less myelosuppression in rats than Taxol. In an acute
toxicity study in rats, cerebral cortical necrosis or severe
neurotoxicity was observed in animals receiving Taxol at 9 mg/kg
but was absent in animals receiving a pharmaceutical composition
comprising paclitaxel and albumin at a dose of up to 120 mg/kg.
Thus the presence of albumin in a pharmaceutical composition
comprising paclitaxel results in a substantial reduction in side
effects and toxicity when compared to conventional pharmaceutical
compositions comprising paclitaxel.
EXAMPLE 17
[0071] This example demonstrates the clinical effects of a
pharmaceutical composition comprising paclitaxel and albumin in
humans.
[0072] Clinical studies in over 500 human patients to date provide
evidence supporting the reduction in toxicity and side-effects for
a pharmaceutical composition comprising paclitaxel and albumin
("albumin-paclitaxel") when compared with cremophor-paclitaxel
compositions (Taxol). In a phase I study of 19 patients, the
maximum tolerated dose of albumin-paclitaxel given every 3 weeks
was 300 mg/m.sup.2. This is substantially higher than the generally
administered dose of cremophor-paclitaxel which is 175 mg/m.sup.2
given once every 3 weeks. The hematological toxicities in these
patients were mild with no hypersensitivities, mild neuropathies,
and no administration related side effects such as venous
irritation, etc.
[0073] In another phase I study of 27 patients, the maximum
tolerated dose of albumin-paclitaxel given on a weekly schedule was
125-150 mg/m.sup.2. This is substantially higher than the generally
administered dose of cremophor-paclitaxel which is 80 mg/m.sup.2
when given on a weekly schedule. The hematological toxicities in
these patients were mild with no hypersensitivities, mild
neuropathies, and no administration related side effects such as
venous irritation, etc.
[0074] In two phase II studies of albumin-paclitaxel given at
either 175 or 300 mg/m.sup.2 every 3 weeks in 43 and 63 patients
respectively, hematological toxicities were low with only 7% and
24% of patients with ANC <500/mm.sup.3 at 175 mg/m.sup.2 and 300
mg/m.sup.2 respectively. Severe neuropathy occurred in 0% and 14%
of patients for 175 mg/m.sup.2 and 300 mg/M2 respectively. There
was no incidence of severe hypersensitivity, and no incidence of
administration related side effects such as venous irritation, pain
on injection, etc. These side effects were substantially lower than
experienced with Taxol.
[0075] In phase III trials comparing the albumin-paclitaxel
composition ABI-007 against Taxol (which contains
cremophor-paclitaxel), the dose of ABI-007 was substantially higher
(260 mg/m.sup.2 vs. 175 mg/m.sup.2 for Taxol) indicating it was
better tolerated. The albumin-paclitaxel compositions also
demonstrated significantly reduced neutropenia when compared to
cremophor-paclitaxel.
EXAMPLE 18
[0076] This example demonstrates enhanced preclinical efficacy
using a pharmaceutical composition comprising albumin and
paclitaxel.
[0077] An in vitro cytotoxicity study comparing the effect of
albumin-paclitaxel and Taxol on cervical squamous cell carcinoma
A431 showed an approximately 4-fold increase in cytotoxic activity
for albumin-paclitaxel with an IC.sub.50 of 0.0038 and 0.012
.mu.g/ml for albumin-paclitaxel and Taxol respectively.
[0078] In five different human xenograft tumor models in athymic
mice (MX-1 mammary, NCI-H522 lung, SK-OV-3 ovarian, PC-3 prostate,
and HT-29 colon), the MTD or equitoxic dose of ABI-007 was
1.5-3.4-fold higher than for Taxol, and resulted in statistically
significant improvement in tumor growth delay (p<0.05) in all
tumors except the lung tumor (p=0.15).
[0079] In the MX 1 mammary model, one hundred percent (100%) of
albumin-paclitaxel treated animals survived 103 days, as compared
to 20-40% surviving in groups treated with equivalent doses of
Taxol.
EXAMPLE 19
[0080] This example demonstrates enhanced clinical efficacy using a
pharmaceutical composition comprising albumin and paclitaxel
administered intra-arterially.
[0081] In a Phase I/II Study of intra-arterial administration of a
pharmaceutical composition comprising albumin and paclitaxel, as
described herein, patients were enrolled for head & neck cancer
(N=31) and cancer of the anal canal (N=12). The dose escalated from
120-300 mg/m.sup.2 given over 30 minutes by percutaneous
superselective intra-arterial infusion, q 3-4wk. Head and neck
cancer patients exhibited a response rate of 76% (N=29), while
patients with cancer of the anal canal exhibited a response rate
64% (N=11).
EXAMPLE 20
[0082] This example demonstrates the preparation of a
pharmaceutical composition containing 3% oil and comprising
propofol and albumin.
[0083] An oil-in-water emulsion containing 1% (by weight) of
propofol was prepared as follows. The aqueous phase was prepared by
adding glycerol (2.25% by weight) and human serum albumin (0.5% by
weight) into water for injection and stirred until dissolved. The
aqueous phase was passed through a filter (0.2 um filter). The oil
phase was prepared by dissolving egg lecithin (0.4% by weight) and
propofol (1% by weight) into soybean oil (3% by weight) at about
50.degree. C.-60.degree. C. and was stirred until dissolved. The
oil phase was added to the aqueous phase and homogenized at
10,000RPM for 5 min. The crude emulsion was high pressure
homogenized at 20,000 psi and recirculated for 15 cycles at
5.degree. C. Alternately, discrete passes through the homogenizer
were used. The final emulsion was filtered (0.2 .mu.m filter) and
stored under nitrogen. The resulting pharmaceutical composition
contained the following general ranges of components (weight %):
propofol 0.5-5%; human serum albumin 0.5-3%; soybean oil 0.5-3.0%;
egg lecithin 0.12-1.2%; glycerol 2.25%; water for injection q.s. to
100; pH 5-8. Suitable chelators, e.g., deferoxamine (0.001-0.1%),
were optionally added.
EXAMPLE 21
[0084] This example demonstrates the preparation of a
pharmaceutical composition containing 5% oil and comprising
propofol and albumin.
[0085] An oil-in-water emulsion containing 1% (by weight) of
propofol was prepared as follows. The aqueous phase was prepared by
adding glycerol (2.25% by weight) and human serum albumin (0.5% by
weight) into water for injection and was stirred until dissolved.
The aqueous phase was passed through a filter (0.2 um filter). The
oil phase as prepared by dissolving egg lecithin (0.8% by weight)
and propofol (1% by weight) into soybean oil (5% by weight) at
about 50.degree. C.-60.degree. C. and was stirred until dissolved.
The oil phase was added to the aqueous phase and homogenized at
10,000RPM for 5 min. The crude emulsion was high pressure
homogenized at 20,000 psi and recirculated for 15 cycles at
5.degree. C. Alternately, discrete passes through the homogenizer
were used. The final emulsion was filtered (0.2 .mu.m filter) and
stored under nitrogen. The resulting pharmaceutical composition
contained the following general ranges of components (weight %):
propofol 0.5-5%; human serum albumin 0.5-3%; soybean oil 0.5-10.0%;
egg lecithin 0.12-1.2%; glycerol 2.25%; water for injection q.s. to
100; pH 5-8. Suitable chelators, e.g., deferoxamine (0.001-0.1%),
were optionally added
EXAMPLE 22
[0086] This example demonstrates the preparation of a
pharmaceutical composition comprising propofol and albumin that is
free of oil.
[0087] Using the procedure similar to that described in Example 18,
propofol compositions containing albumin and tween 80 were
prepared. The aqueous phase was prepared by adding glycerol (2.25%
by weight), human serum albumin (0.5% by weight), tween 80 (1.5% by
weight) and deferoxamine mesylate (0.1% by weight) into water for
injection and stirred until dissolved. The aqueous phase was passed
through a filter (0.2 .mu.m filter). Propofol (1% by weight) was
added to the aqueous phase and homogenized at 10,000 RPM for 5 min.
The crude emulsion was high pressure homogenized at 20,000 psi and
recirculated for 15 cycles at 5.degree. C. Alternately, discrete
passes through the homogenizer were used. The final emulsion was
filtered (0.2 um filter) and stored under nitrogen. The resulting
pharmaceutical composition contained the following general ranges
of components (weight %): propofol 0.5-5; human serum albumin
0.5-3%; tween 80 0.1-1.5%; deferoxamine mesylate 0.0001-0.1%;
glycerol 2.25%; water for injection q.s. to 100; pH 5-8.
EXAMPLE 23
[0088] This example demonstrates the preparation of a
pharmaceutical composition comprising propofol, albumin, and
vitamin E-TPGS, which is free of oil.
[0089] Using the procedure similar to that described in Example 19,
propofol compositions containing albumin and vitamin E-TPGS were
prepared. The aqueous phase was prepared by adding glycerol (2.25%
by weight), human serum albumin (0.5% by weight), vitamin E-TPGS
(1% by weight) and deferoxamine mesylate (0.1% by weight) into
water for injection and was stirred until dissolved. The aqueous
phase was passed through a filter (0.2 um filter). Propofol (1% by
weight) was added to the aqueous phase and homogenized at 10,000
RPM for 5 min. The crude emulsion was high pressure homogenized at
20,000 psi and recirculated for 15 cycles at 5.degree. C.
Alternately, discrete passes through the homogenizer were used. The
final emulsion was filtered (0.2 .mu.m filter) and stored under
nitrogen. The resulting pharmaceutical composition contained the
following general ranges of components (weight %): propofol 0.5-5;
human serum albumin 0.5-3%; vitamin E-TPGS 0.5-4.0%; optionally
deferoxamine mesylate 0.0001-0.1%; glycerol 2.25%; water for
injection q.s. to 100; pH 5-8.
EXAMPLE 24
[0090] This example demonstrates the preparation of a
pharmaceutical composition comprising propofol, albumin, vitamin
E-TPGS, and 1% oil.
[0091] An emulsion containing 1% (by weight) of propofol was
prepared by the following method. The aqueous phase was prepared by
adding glycerol (2.25% by weight) and human serum albumin (0.5% by
weight) into water for injection and stirred until dissolved. The
aqueous phase was passed through a filter (0.2 .mu.m filter).
Surfactant, e.g., Vitamin E-TPGS (0.5%), was added to aqueous
phase. The oil phase consisted of propofol (1% by weight) and 1%
soybean oil. The oil phase was added to the aqueous phase and
homogenized at 10,000 RPM for 5 min. The crude emulsion was high
pressure homogenized at 20,000 psi and recirculated for up to 15
cycles at 5.degree. C. Alternatively, discrete passes through the
homogenizer were used. The final emulsion was filtered (0.2 .mu.m
filter) and stored under nitrogen.
[0092] The resulting pharmaceutical composition contained the
following general ranges of components (weight %): propofol 0.5-5%;
human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; soybean or
other oil (0.1%-5%); glycerol 2.25%; water for injection q.s. to
100; pH 5-8. Deferoxamine was optionally added (0.001%-0.1% by
weight).
EXAMPLE 25
[0093] This example demonstrates the preparation of a
pharmaceutical composition comprising propofol, albumin, vitamin
E-TPGS, 1% oil, and a negatively charged component.
[0094] An emulsion containing 1% (by weight) of propofol was
prepared by the following method. The aqueous phase was prepared by
adding glycerol (2.25% by weight) and human serum albumin (0.5% by
weight) into water for injection and was stirred until dissolved.
The aqueous phase was passed through a filter (0.2 .mu.m filter).
Surfactant, e.g., Vitamin E-TPGS (0.5%), was added to aqueous
phase. The oil phase consisted of propofol (1% by weight) and 1%
soybean oil. A small quantity of negatively charged component
(0.001%-1%), e.g., a phospholipid or bile salt was added. The oil
phase was added to the aqueous phase and homogenized at 10,000 RPM
for 5 min. The crude emulsion was high pressure homogenized at
20,000 psi and recirculated for up to 15 cycles at 5.degree. C.
Alternatively, discrete passes through the homogenizer were used.
The final emulsion was filtered (0.2 .mu.m filter) and stored under
nitrogen.
[0095] The resulting pharmaceutical composition contained the
following general ranges of components (weight %): propofol 0.5-5%;
human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; soybean or
other oil (0.1%-5%); glycerol 2.25%; water for injection q.s. to
100; pH 5-8. Deferoxamine was optionally added (0.001%-0.1% by
weight).
EXAMPLE 26
[0096] This example demonstrates the preparation of a
pharmaceutical composition comprising propofol, albumin, vitamin
E-TPGS, 1% oil, and a negatively charged component (sodium
deoxycholate).
[0097] An emulsion containing 1% (by weight) of propofol was
prepared by the following method. The aqueous phase was prepared by
adding glycerol (2.25% by weight) and human serum albumin (0.5% by
weight) into water for injection and stirred until dissolved. The
aqueous phase was passed through a filter (0.2 .mu.m filter).
Surfactant, e.g., Vitamin E-TPGS (0.5%), was added to aqueous
phase. The oil phase consisted of propofol (1% by weight) and 1%
soybean oil. A small quantity of negatively charged component
(0.001%-1%), e.g., sodium deoxycholate was added. The oil phase was
added to the aqueous phase and homogenized at 10,000 RPM for 5 min.
The crude emulsion was high pressure homogenized at 20,000 psi and
recirculated for up to 15 cycles at 5.degree. C. Alternately,
discrete passes through the homogenizer were used. The final
emulsion was filtered (0.2 .mu.m filter) and stored under
nitrogen.
[0098] The resulting pharmaceutical composition contained the
following general ranges of components (weight %): propofol 0.5-5%;
human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; soybean or
other oil (0.1%-5%); glycerol 2.25%; water for injection q.s. to
100; pH 5-8. Deferoxamine was optionally added (0.001%-0.1% by
weight).
EXAMPLE 27
[0099] This example demonstrates the preparation of a
pharmaceutical composition comprising propofol, albumin, vitamin
E-TPGS, 1% oil, and a negatively charged component (phospholipids,
bile salts, polyaminoacids etc).
[0100] An emulsion containing 1% (by weight) of propofol was
prepared as follows. The aqueous phase was prepared by adding
glycerol (2.25% by weight) and human serum albumin (0.5% by weight)
into water for injection and stirred until dissolved. The aqueous
phase was passed through a filter (0.2 .mu.m filter). Surfactant,
e.g., Vitamin E-TPGS (0.5%), was added to aqueous phase. The oil
phase consisted of propofol (1% by weight) and 1% soybean oil. A
small quantity of negatively charged component (0.001%-1%), e.g.,
phosphatidyl choline was added. The oil phase was added to the
aqueous phase and homogenized at 10,000 RPM for 5 min. The crude
emulsion was high pressure homogenized at 20,000 psi and
recirculated for up to 15 cycles at 5.degree. C. Alternatively,
discrete passes through the homogenizer were used. The final
emulsion was filtered (0.21 .mu.m filter) and stored under
nitrogen.
[0101] The resulting pharmaceutical composition contained the
following general ranges of components (weight %): propofol 0.5-5%;
human serum albumin 0.01-3%; Vitamin E-TPGS 0.1-2%; soybean or
other oil (0.1%-5%); glycerol 2.25%; water for injection q.s. to
100; pH 5-8. Deferoxamine was optionally added (0.001%-0.1% by
weight).
EXAMPLE 28
[0102] This example demonstrates the binding of propofol to
albumin.
[0103] The binding of propofol to albumin was determined as
follows. Solubility of propofol was tested in water and in
solutions containing albumin. 250 .mu.L of propofol was added to 10
mL of a water or albumin solution and stirred for 2 hours in a
scintillation vial. The solution was then transferred to a 15 mL
polyethylene centrifuge tube and kept at 40.degree. C. for about 16
hours. Samples of water and albumin solutions were assayed for
propofol. Solubility of propofol in water was determined to be 0.12
mg/ml. Solubility of propofol in albumin solutions was dependent on
the concentration of albumin and increased to 0.44 mg/ml when the
albumin concentration was 2% (20 mg/ml). The solutions were
ultrafiltered through a 30 kD MWCO filter and the filtrates were
assayed for propofol. It was found that for the propofol/water
solution, 61% of the propofol could be recovered in the filtrate
whereas for the propofol/albumin solution, only 14% was recovered
in the filtrate, indicating a substantial binding of propofol with
albumin. Based on these results, addition of albumin to
pharmaceutical compositions comprising propofol result in a
decrease in the amount of free propofol due to albumin binding of
the propofol.
EXAMPLE 29
[0104] This example demonstrates the reduction of free propofol in
a pharmaceutical composition by filtration/membrane contact.
[0105] As observed in the experiments described in Example 28,
filtration or ultrafiltration of pharmaceutical compositions
comprising propofol results in a reduction in the amount of free
propofol. Diprivan and a pharmaceutical composition prepared in
accordance with the present invention containing albumin, each of
which contained 1% propofol (10 mg/ml), were ultrafiltered using a
30 kD membrane. The amount of free propofol was measured in the
filtrate using HPLC. The concentration of free propofol in the
filtrate was about 17 .mu.g/ml for Diprivan, while the
concentration of free propofol in the filtrate was about 7 .mu.g/ml
for the inventive pharmaceutical composition. The results
correspond to an effective reduction of free propofol by greater
than a factor of 2 for pharmaceutical composition comprising
propofol and albumin.
EXAMPLE 30
[0106] This example demonstrates administration of a pharmaceutical
composition comprising propofol and albumin to humans.
[0107] A randomized, double-blind clinical trial was conducted to
compare adverse skin sensations of a pharmaceutical composition
comprising propofol and albumin with that of a commercially
available propofol formulation, Diprivan. Trials were conducted in
compliance with Good Clinical Practices and informed consent was
taken from the subjects.
[0108] Adult human subjects of either sex were eligible for
participation if they had unbroken, apparently normal skin of
dorsal side of their hands.
[0109] The formulations originally stored in a refrigerator were
brought to room temperature and then 10 .mu.L of the formulations
was placed slowly on the back side of both the hands of a subject
simultaneously. The overall reaction and feel on their hands for
the formulations were noted. The results of this study are set
forth in Table 1.
1TABLE 1 % of subjects with ABI- % of subjects with Diprivan Order
of Propofol sensation sensation a test on a Mild warm or No Mild
warm or No subject stinging or biting sensation stinging or biting
sensation 1st 0.0 100.0 75 25 incidence
EXAMPLE 31
[0110] This example demonstrates the use of deferoxamine as
antioxidant in a pharmaceutical composition comprising
propofol.
[0111] Pharmaceutical compositions comprising propofol and
deferoxamine mesylate, and containing tween or TPGS were stored at
4.degree., 25.degree., or 40.degree. C. to test the effect of
deferoxamine mesylate in preventing oxidation of propofol. The
concentration of propofol was measured for these formulations over
time to determine the antioxidant activity of deferoxamine. The
data is reported below in Tables 2 and 3 as % potency relative to
time zero.
2TABLE 2 Albumin/tween formulation 1 month Storage Temp 4.degree.
C. 25.degree. C. 40.degree. C. CONTROL 100% 88% 48% 0.01% Def 101%
89% 61% 0.1% Def 103% 89% 64%
[0112]
3TABLE 3 Albumin/TPGS formulation 1 month Storage Temp 4.degree. C.
25.degree. C. 40.degree. C. CONTROL 99% 73% 42% 0.01% DEF 99% 87%
55% 0.1% DEF 99% 85% 58%
[0113] Under these conditions, deferoxamine was efficient in
reducing the level of oxidation of propofol. The effect was more
pronounced at higher temperatures. No significant oxidation
occurred at 4.degree. C. This study was conducted using stoppers
that were not inert or Teflon coated.
EXAMPLE 32
[0114] This example demonstrates intrapulmonary delivery of a
pharmaceutical composition comprising paclitaxel and albumin
(ABI-007).
[0115] The purpose of this study was to determine the time course
of [.sup.3H]ABI-007 in blood and select tissues following
intratracheal instillation to Sprague Dawley rats.
[0116] The target volume of the intratracheal dose formulation to
be administered to the animals was calculated based on a dose
volume of 1.5 mL per kg body weight. The dosing apparatus consisted
of a Penn-Century microsprayer (Model 1A-1B; Penn-Century, Inc.,
Philadelphia, Pa.; purchased from DeLong Distributors, Long Branch,
N.J.) attached to a 1-mL gas-tight, luer-lock syringe. The
appropriate volume of dose preparation was drawn into the dosing
apparatus, the filled apparatus was weighed and the
weight-recorded. A catheter was placed in the trachea of the
anesthetized animal, the microsprayer portion of the dosing
apparatus was placed into the trachea through the catheter, and the
dose was administered. After dose administration the empty dosing
apparatus was reweighed and the administered dose was calculated as
the difference in the weights of the dosing apparatus before and
after dosing. The average dose for all animals was 4.7738.+-.0.0060
(CV 1.5059) mg paclitaxel per kg body weight.
[0117] Blood samples of approximately 250 .mu.L were collected from
the indwelling jugular cannulas of JVC rats at the following
predetermined post-dosing time points: 1, 5, 10, 15, 30, and 45
minutes (min), and 1, 4, 8, and 24 hours (h). The 24-h blood
samples, as well as blood samples collected from animals sacrificed
at 10 min, 45 min, and 2 h, were collected via cardiac puncture
from anesthetized rats at sacrifice. All blood samples analyzed for
total radioactivity were dispensed into pre-weighed sample tubes,
and the sample tubes were reweighed, and the weight of each sample
was calculated by subtraction. The blood samples collected from the
jugular vein as well as the 250-.mu.L aliquots of blood collected
from each animal at sacrifice were assayed for total tritium
content.
[0118] For all rats, the maximum concentration of tritium in blood
was observed at 5 min (0.0833 hr) post dosing. The elimination
half-life of tritium, determined over the time interval from 4 h to
24 h, ranged from 19.73 h to 43.02 h. It should be noted that this
interval includes only three data points, which may account for the
variability in this parameter. The apparent clearance of tritium
from blood was on the order of 0.04 L/h. The results of these
experiments are set forth below in Table 4.
4TABLE 4 Noncompartmental Analysis of Blood Tritium Concentration
(mg-eq/L) vs. Time Profiles in Rats After Intratracheal
Instillation of [.sup.3H]ABI-007 Parameter Mean +/- SD C.sub.max
(mg-eq/L) 1.615 +/- 0.279 T.sub.max (hr) 0.0833 +/- 0.0 t1/2 beta
(hr) 33.02 +/- 1.99 AUClast (mg-eq .times. hr/L) 7.051 +/- 1.535
Cl/F (L/hr) 0.0442 +/- 0.0070 Fa (Bioavailability) 1.229 +/-
0.268
[0119] The mean blood concentration of [.sup.3H]ABI-007-derived
radioactivity after an intravenous dose to rats was analyzed as a
function of time in order to evaluate the bioavailability of
tritium derived from an intratracheal dose of [.sup.3H]ABI-007.
This analysis resulted in a 24-hour AUC (AUClast) of 6.1354 mg-eq
.quadrature. hr/L. Based on these data, radioactivity derived from
the intratracheal dose of [.sup.3H]ABI-007 is highly bioavailable.
These analyses are based on total radioactivity.
[0120] Tritium derived from [.sup.3H]ABI-007 is rapidly absorbed
after intratracheal instillation. The average absorption and
elimination half-lives (k01 half-life and k10 half-life,
respectively) for tritium in blood after an intratracheal dose of
[.sup.3H]ABI-007 (mean +/-SD) were 0.0155+/-0.0058 hr and
4.738+/-0.366 hr, respectively. The average apparent clearance of
tritium from blood was 0.1235+/-0.0180 L/hr (see Table 4
above).
[0121] Tritium derived from [.sup.3H]ABI-007 was absorbed and
distributed after intratracheal administration. The time course of
tritium in blood was well described by a two-compartment model,
with mean absorption and elimination half-lives of 0.0155 and 4.738
hr, respectively. Approximately 28% of the administered dose was
recovered in the lung at 10 min after the intratracheal dose. A
maximum of less than 1% of the dose was recovered in other tissues,
excluding the gastrointestinal tract, at all time points
examined.
[0122] Based on results from a previously conducted intravenous
dose study with [.sup.3H]Capxol.TM., the bioavailability of tritium
derived from the intratracheal dose was 1.229.+-.0.268 (mean
.+-.SD) for the three animals in this dose group. It should be
noted, however, that this estimate of bioavailability is based on
total radioactivity. Surprisingly, paclitaxel delivered by the
pulmonary route using invention compositions with albumin was
rapidly bioavailable indicating excellent transport across
pulmonary endothelium. No toxicity in the animals was noted, which
was surprising since pulmonary delivery of cytotoxics is known to
cause lung toxicities.
[0123] A fair amount of radioactivity was present in the
gastrointestinal tract (including contents) at 24 hr post dosing
(27% for the intratracheal dose). The presence of tritium in the
gastrointestinal tract may be due to biliary excretion or clearance
of tritium from the respiratory tract via mucociliary clearance
with subsequent swallowing.
EXAMPLE 33
[0124] This example demonstrates an investigation of Aerotech II
and Pari nebulizers for pulmonary delivery of pharmaceutical
compositions comprising paclitaxel and albumin.
[0125] The study was carried out using the paclitaxel-albumin
pharmaceutical composition ABI-007 under the following conditions:
room temperature (20-23.degree. C.), relative humidity (48-54%),
ambient pressure (629 mmHg), nebulizer flowrate (10 L/min for
Aerotech II; 7 L/min for Pari), total flowrate (28.3 L/min),
nebulizer pressure drop (23 lb/in.sup.2 for Aerotech II; 32 lb/in2
for Pari), run time (15 to 60 seconds), sample volume (1.5 mL),
ABI-007 paclitaxel concentration (5,10, 15 and 20 mg/mL).
[0126] Both Aerotech II and Pari nebulizers provided acceptable
overall efficiency (30%-60%) when ABI-007 was reconstituted at a
concentration range of 5-15 mg/mL. The Pari nebulizer efficiency
had higher nebulizer efficiency than the Aerotech II nebulizer. The
Pari nebulizer efficiency decreased somewhat as ABI-007
concentration increased. Excellent fine particle fraction was
observed (74%-96%). The Aerotech II nebulizer had higher fine
particle fraction than the Pari nebulizer. The fine particle
fraction was independent of concentration.
[0127] The Pari nebulizer delivered 100 mg of paclitaxel in less
than 30 minutes using a 15 mg/mL solution of ABI-007. The Aerotech
II nebulizer delivered 100 mg of paclitaxel in about 65 min using
either a 10 mg/mL or 15 mg/mL solution of ABI-007. Performance
stability was tested for both Aerotech II and Pari nebulizers.
Aerosol concentration and efficiency of both nebulizers were stable
until the drug was exhausted. At 15 mg/mL, the Pari nebulizer
consumed the drug at twice the rate of the Aerotech II nebulizer
and produced higher aerosol concentrations than that of the
Aerotech II nebulizer.
[0128] In conclusion, the nanoparticle/albumin formulation of
paclitaxel (ABI-007) shows excellent bioavailability in rats when
administered by the pulmonary route. There were no overt signs of
early toxicity at the administered dose. Pulmonary delivery of
nanoparticle paclitaxel (ABI-007) may be achieved using
conventional nebulizers.
EXAMPLE 34
[0129] This example describes intrapulmonary delivery of a
pharmaceutical composition comprising albumin and rapamycin. The
purpose of this study was to determine the pulmonary absorption of
rapamycin in blood following intratracheal instillation to Sprague
Dawley rats as compared to intravenous installation.
[0130] The target volume of the intratracheal dose formulation that
was administered to the animals was calculated based on a dose
volume of 1 mL per kg body. The intratracheal dosing apparatus
consisted of a Penn-Century microsprayer (Model 1A-1B;
Penn-Century, Inc., Philadelphia, Pa.; purchased from DeLong
Distributors, Long Branch, N.J.) attached to a 1 mL gas-tight,
luer-lock syringe. The appropriate volume of dose preparation was
drawn into the dosing apparatus, the filled apparatus was weighed
and the weight-recorded. A catheter was placed in the trachea of
the anesthetized animal, the microsprayer portion of the dosing
apparatus was placed into the trachea through the catheter, and the
dose was administered. After dose administration the empty dosing
apparatus was reweighed and the administered dose was calculated as
the difference in the weights of the dosing apparatus before and
after dosing.
[0131] 250 .mu.L samples were collected from the indwelling jugular
cannulas of rats at the following predetermined post-dosing time
points: 1, 5, 10, 15, 30, and 45 minutes (min) and 1, 4, 8, and 24
hours (h). All blood samples analyzed were dispensed into
pre-weighed sample tubes, and the sample tubes were reweighed, and
the weight of each sample was calculated by subtraction. The blood
samples collected were assayed for total rapamycin concentration
using LC/MS/MS.
[0132] Surprisingly, the results showed no significant difference
in the blood concentration of rapamycin delivered via pulmonary
route versus intravenously. The bioavailability of rapamycin
delivered by the pulmonary route using a pharmaceutical composition
comprising albumin was calculated to be 109%, indicating excellent
transport across pulmonary endothelium.
EXAMPLE 35
[0133] This example demonstrates tissue distribution of
albumin-rapamycin after intrapulmonary administration of a
pharmaceutical composition comprising rapamycin and albumin
prepared in accordance with the present invention. The purpose of
this study was to determine the pulmonary absorption of rapamycin
in tissue following intratracheal instillation to Sprague Dawley
rats as compared to intravenous installation.
[0134] The target volume of the intratracheal dose formulation that
was administered to the animals was calculated based on a dose
volume of 1 mL per kg body. The dosing apparatus consisted of a
Penn-Century microsprayer (Model 1A-1B; Penn-Century, Inc.,
Philadelphia, Pa.; purchased from DeLong Distributors, Long Branch,
N.J.) attached to a 1-mL gas-tight, luer-lock syringe. The
appropriate volume of dose preparation was drawn into the dosing
apparatus, the filled apparatus was weighed and the
weight-recorded. A catheter was placed in the trachea of the
anesthetized animal, the microsprayer portion of the dosing
apparatus was placed into the trachea through the catheter, and the
dose was administered. After dose administration the empty dosing
apparatus was reweighed and the administered dose was calculated as
the difference in the weights of the dosing apparatus before and
after dosing.
[0135] Samples were collected from the brain, lung, and, liver of
three rats per group per time point at 10 minutes, 45 minutes, 2
hours, and 24 hours. The samples were collected and analyzed for
total rapamycin concentration using LC/MS/MS. The results indicate
that rapamycin concentration is greater in lung tissue when
delivered via pulmonary as compared to intravenous delivery.
However, the total concentration of rapamycin in the brain is lower
when delivered via intratracheal (IT) as compared to intravenous
(IV). In the liver, there appears to be no difference in the
concentration of rapamycin whether delivered IT or IV. Based on
these results, pulmonary delivery of rapamycin may be suitable for
the treatment of a condition (i.e., lung transplantation), wherein
high local concentration of rapamycin would be beneficial.
EXAMPLE 36
[0136] This example demonstrates oral delivery of a pharmaceutical
composition comprising paclitaxel and albumin (ABI-007).
[0137] Tritiated ABI-007 was utilized to determine oral
bioavailability of paclitaxel following oral gavage in rats.
Following overnight fasting, 5 rats were given 5.5 mg/kg paclitaxel
in ABI-007 (Group A) and another 5 rats (Group B) were pretreated
with cyclosporine (5.0 mg/kg) followed by 5.6 mg/kg paclitaxel in
ABI-007. A pharmacokinetic analysis of blood samples drawn at 0.5,
1, 2, 3, 4, 5, 6, 8, 12, and 24 hours was performed after
determination of radioactivity in the blood samples by combustion.
Oral bioavailability was determined by comparison with intravenous
data previously obtained. The results are set forth below in Table
5.
5TABLE 5 Mean AUC 0-24, C.sub.max, T.sub.max and % Absorption of
.sup.3H-Paclitaxel Derived Radioactivity Following Oral
Administration Dose/Route AUC0-24 Absorption Cmax (mg/kg) Tmax
Group Treatment mg/kg (.mu.g eq .times. hr/mL) (%) (.mu.g .times.
eq/mL) (hr) A ABI-007 in 5.5/PO(P) 2.92 44.3 0.245 1 Normal Saline
B ABI-007 in 5/PO(C), 5.6/PO(P) 8.02 121.1 0.565 0.5 Normal Saline
with CsA
[0138] AUC 0-24 IV (6.06 .mu.g.times.hr./mL) and IV dose (5.1
mg/kg) were used for calculation of percent absorption (data based
on IV dose of ABI-007).
[0139] An oral bioavailability of 44% was seen for ABI-007 alone.
This is dramatically higher than is seen for other formulations of
paclitaxel. The bioavailability increased to 121% when animals were
treated with cyclosporine (CsA). This is expected as CsA is a known
suppressor of the p-glycoprotein pump that would normally prevent
absorption of compounds such as paclitaxel from the GI tract. The
greater than 100% bioavailability can be explained by reabsorption
following biliary excretion of paclitaxel into the GI tract. Other
known suppressors or enhancers of absorption may be also utilized
for this purpose.
EXAMPLE 37
[0140] This example demonstrates improved penetration of paclitaxel
into red blood cells and tumor cells upon administration of a
pharmaceutical composition comprising paclitaxel and albumin.
[0141] Human MX-1 breast tumor fragments were implanted
subcutaneously in athymic mice. A pharmaceutical composition
comprising paclitaxel and albumin ("paclitaxel-albumin"), as
described previously, and Taxol were prepared with .sup.3H
paclitaxel to a specific activity of 25 .mu.Ci/mg paclitaxel. 20
mg/kg radiolabeled paclitaxel-albumin or Taxol was administered
intravenously in saline when tumor volume reached approximately 500
mm.sup.3. Plasma, blood, and tumor tissue were sampled and analyzed
for radioactivity at 5, 15, and 30 minutes and at 1, 3, 8, and 24
hours after administration. Tumor pharmacokinetic (AUC and
absorption constant) was analyzed using WinNonlin, Pharsight,
USA.
[0142] Paclitaxel-albumin exhibited rapid partitioning into red
blood cells (RBCs) as shown by a rapid drop of the plasma/blood
radioactivity ratio to unity after intravenous administration of
the drug. Complete partitioning into RBCs occurred as early as 1 hr
after administration of paclitaxel-albumin. In contrast, the
partitioning of paclitaxel formulated as Taxol into RBCs was much
slower and was not completed until more than 8 hrs.
[0143] Paclitaxel-albumin exhibited a rapid partitioning into tumor
tissue with an absorption constant (K.sub.a) that was 3.3.times.
greater than Taxol. The K.sub.a were 0.43 hr.sup.-1 and 0.13
hr.sup.-1 for paclitaxel-albumin and Taxol, respectively. Rapid
uptake of paclitaxel resulted in 33% higher tumor AUC for
paclitaxel-albumin than for Taxol. The AUC were 3632 nCi*hr/g and
2739 nCi*hr/g for paclitaxel-albumin and Taxol, respectively.
EXAMPLE 38
[0144] This example demonstrates the safety of a pharmaceutical
composition comprising paclitaxel and albumin administered to
mice.
[0145] Athymic mice were treated with escalating doses of
paclitaxel-albumin or Taxol everyday for 5 consecutive days.
Survival was plotted versus dose to determine the LD.sub.50.
Survival was greatly improved with paclitaxel-albumin versus Taxol
(p=0.017, ANOVA). The LD.sub.50 for paclitaxel-albumin and Taxol
were calculated to be 47 mg/kg/day and 30 mg/kg/day for a
qld.times.5 schedule, respectively. At a dose level of 13.4
mg/kg/day, both paclitaxel-albumin and Taxol were well tolerated
with mortality of 1% (1 death out of 72 mice) and 4% (2 deaths out
of 47 mice), respectively. At a dose level of 20 mg/kg/day, there
was 1% mortality for paclitaxel-albumin (1 death out of 72 mice)
versus 17% mortality for Taxol (8 deaths out of 47 mice)
(p=0.0025). At a dose level of 30 mg/kg/day, there was 4% mortality
for paclitaxel-albumin (3 deaths out of 72 mice) versus 49%
mortality for Taxol (23 deaths out of 47 mice) (p<0.0001).
EXAMPLE 39
[0146] This example demonstrates a novel paclitaxel transport
mechanism across microvessel endothelial cells (EC) for
paclitaxel-albumin compositions.
[0147] Nanoparticles and albumin-paclitaxel compositions can
accumulate in tumor tissue due to EPR effect resulting from `leaky`
vessels in a tumor. An albumin specific gp60 receptor (albondin)
transported albumin across EC by transcytosis of the receptors
within caveolae at the cell surface. This transcytosis mechanism
allows for the transport of albumin-paclitaxel to the underlying
interstitial space. In contrast, cremophor in Taxol inhibited
binding of paclitaxel to albumin, greatly reducing paclitaxel
transport to the tumor. In addition, the gp16 and gp30 receptors
also were involved in intracellular transport of modified albumins
containing bound paclitaxel, resulting in increased binding of
paclitaxel to endothelial cells with a greater anti-angiogenic
effect as compared to Taxol.
EXAMPLE 40
[0148] This example demonstrates an increase in endothelial
transcytosis of pharmaceutical compositions comprising paclitaxel
and albumin as compared to Taxol.
[0149] Human lung microvessel endothelial cells (HLMVEC) were grown
to confluence on a transwell. The inventive pharmaceutical
composition comprising paclitaxel and albumin, or Taxol containing
fluorescent paclitaxel (Flutax) at a concentration of 20 .mu.g/mL,
was added to the upper transwell chamber.
[0150] The transport of paclitaxel by transcytosis from the upper
chamber to the lower chamber was monitored continuously using a
fluorometer. A control containing only Flutax without albumin was
also used. The control with Flutax showed no transport, validating
the integrity of the confluent HLMVEC monolayer. Transport of
paclitaxel from the albumin-paclitaxel composition was much faster
than paclitaxel from Taxol in the presence of 5% HSA (physiological
concentration). Transport rate constants (K.sub.t) for the
albumin-paclitaxel composition and Taxol were 1.396 hr.sup.-1 and
0.03 hr.sup.-1, respectively. The total amount of paclitaxel
transported across the monolayer was three times higher for the
albumin-paclitaxel composition than Taxol.
EXAMPLE 41
[0151] This example demonstrates improved endothelial cell (EC)
binding by pharmaceutical compositions comprising paclitaxel and
albumin as compared to Taxol.
[0152] Human umbilical vein endothelial cells (HUVEC) were grown on
a 96-well microtiter plate. In one experiment, paclitaxel
(Flutax-Oregon Green labeled paclitaxel) was reacted with the HUVEC
in the presence of increasing concentrations of Cremophor EL/EtOH,
which is the vehicle for Taxol. In another experiment, a
pharmaceutical composition comprising albumin and Flutax and a
Taxol-Flutax composition were reacted to the HUVEC at various final
concentrations. Binding of paclitaxel to cells was inhibited by
Cremophor. Inhibition was exhibited by an IC.sub.50 of 0.02% of
Cremophor EL/EtOH. This concentration of Cremophor has been shown
to persist during Taxol chemotherapy for at least 24 hours.
Therefore, it is a relevant process in vivo. At all concentrations
tested, a significant amount of paclitaxel from the
albumin-paclitaxel composition became bound to cells. In
comparison, little or no binding was observed for Taxol.
EXAMPLE 42
[0153] This example demonstrates improved albumin binding by
pharmaceutical compositions comprising paclitaxel and albumin as
compared to Taxol.
[0154] Human Serum Albumin (HSA) was immobilized on a plastic ELISA
plate. Paclitaxel (Flutax-Oregon Green labeled paclitaxel) was
reacted with the immobilized HSA in presence of increasing
concentrations of Cremophor EL/EtOH. In another experiment, an
albumin-paclitaxel-Flutax composition and a Taxol-Flutax
composition were reacted to immobilized HSA at a final
concentration of 20 .mu.g paclitaxel/mL. Binding of paclitaxel to
albumin was inhibited by Cremophor. Inhibition was exhibited by an
IC.sub.50 of 0.003% of Cremophor EL/EtOH. This concentration of
Cremophor has been shown to persist during Taxol chemotherapy for
at least 24 hours. Therefore, it is a relevant process in vivo. At
a relevant pharmacologic paclitaxel concentration (20 .mu.g/mL), a
significant amount of paclitaxel from the albumin-paclitaxel
composition became bound to immobilized HSA. In comparison, no
binding was observed for Taxol.
EXAMPLE 43
[0155] This example demonstrates increased transfer of paclitaxel
to albumin for pharmaceutical compositions comprising paclitaxel
and albumin as compared to Taxol.
[0156] Taxol-Flutax and albumin-paclitaxel-Flutax compositions were
mixed with either 5% HSA in Hanks buffer or serum, at 20 .mu.g/mL,
40 .mu.g/ml, and 80 .mu.g/ml. The mixtures were immediately
separated on a native 3-14% polyacrylamide gel and the amount of
paclitaxel bound to albumin was determined by a scanning
fluorometer. The transfer of paclitaxel to HSA was more rapid for
the albumin-paclitaxel composition versus Taxol. More paclitaxel
co-electrophoresed with HSA when either serum or 5% HSA was
incubated with the albumin-paclitaxel-Flutax composition or the
Taxol-Flutax composition. Upon exposure to 5% HSA, 45%, 60%, and
33% more paclitaxel transferred to HSA for the
albumin-paclitaxel-Flutax composition than for the Taxol-Flutax
composition, at 20 .mu.g/ml, 40 .mu.g/ml, and 80 .mu.g/ml,
respectively. Upon exposure to human serum, 121%, 31%, and 83% more
paclitaxel transferred to HSA for the albumin-paclitaxel-Flutax
composition than for the Taxol-Flutax composition, at 20 .mu.g/ml,
40 .mu.g/ml, and 80 .mu.g/ml, respectively. The C.sub.max for
ABI-007 at 260 mg/m.sup.2 is approximately 20 .mu.g/mL, therefore
this is an important process in vivo.
EXAMPLE 44
[0157] This example demonstrates that the glycoprotein receptor
gp60 is responsible for binding and transcytosis of
albumin-paclitaxel.
[0158] Fluorescent labeled paclitaxel (Flutax) albumin compositions
were contacted with microvessel endothelial cells in culture.
Fluorescent staining was observed under a microscope with evidence
of punctuate areas that were postulated to be the gp60 receptor
binding the albumin-paclitaxel. This was confirmed by using
rhodamine labeled albumin which colocalized with the punctuate
fluorescence of paclitaxel.
EXAMPLE 45
[0159] This example demonstrates that increasing amounts of albumin
can compete with binding of paclitaxel.
[0160] Albumin was immobilized on a microtiter plate. Fluorescent
paclitaxel was added into the wells and the binding of paclitaxel
was measured using a scanning fluorometer. Increasing amounts of
albumin were added to the wells and the level of inhibiton of
paclitaxel binding to immobilized albumin was measured. The data
showed that as the amount of albumin added was increased, a
corresponding decrease in binding was seen. A similar effect was
seen with binding to endothelial cells. This indicated that higher
albumin concentration inhibited binding of paclitaxel. Thus
invention compositions having lower amounts of albumin are
preferred.
EXAMPLE 46
[0161] This example demonstrates that lower amounts of albumin in
the inventive pharmaceutical composition results in stable
compositions.
[0162] To investigate if lower amounts of albumin in compositions
would affect stability of the inventive pharmaceutical composition,
albumin-paclitaxel compositions with low amounts of albumin were
prepared. It was found that these compositions were as stable as
compositions with higher quantities of albumin when examined for
several months at different temperatures (2-8.degree. C.,
25.degree. C. and 40.degree. C.) for potency of paclitaxel,
impurity formation, particle size, pH and other typical parameters
of stability. Thus compositions with lower amounts of albumin are
preferred as this can greatly reduce cost as well as allow
increased binding and transport to cells.
EXAMPLE 47
[0163] This example demonstrates a pharmaceutical composition
comprising albumin and paclitaxel having a high albumin to
paclitaxel ratio.
[0164] 30 mg of paclitaxel was dissolved in 3.0 ml methylene
chloride. The solution was added to 27.0 ml of human serum albumin
solution (3% w/v) (corresponding to a ratio of albumin to
paclitaxel of 27). Deferoxamine was added as necessary. The mixture
was homogenized for 5 minutes at low RPM (Vitris homogenizer, model
Tempest I.Q.) in order to form a crude emulsion, and then
transferred into a high pressure homogenizer (Avestin). The
emulsification was performed at 9000-40,000 psi while recycling the
emulsion for at least 5 cycles. The resulting system was
transferred into a rotary evaporator, and methylene chloride was
rapidly removed at 40.degree. C., at reduced pressure (30 mm Hg)
for 20-30 minutes. The resulting dispersion was translucent, and
the typical average diameter of the resulting paclitaxel particles
was in the range 50-220 nm (Z-average, Malvern Zetasizer). The
dispersion was further lyophilized for 48 hrs. The resulting cake
was easily reconstituted to the original dispersion by addition of
sterile water or saline. The particle size after reconstitution was
the same as before lyophilization.
[0165] It should be recognized that the amounts, types and
proportions of drug, solvents, proteins used in this example are
not limiting in any way. When compared to toxicity of paclitaxel
dissolved in cremophor formulations, the inventive pharmaceutical
composition containing albumin showed substantially lower
toxicity.
EXAMPLE 48
[0166] This example demonstrates a pharmaceutical composition
comprising albumin and paclitaxel having a low albumin to
paclitaxel ratio.
[0167] Specifically, 300 mg of paclitaxel was dissolved in 3.0 ml
methylene chloride. The solution was added to 27 ml of human serum
albumin solution (5% w/v). (corresponding to a ratio of albumin to
paclitaxel of 4.5). Deferoxamine was added as necessary. The
mixture was homogenized for 5 minutes at low RPM (Vitris
homogenizer, model Tempest I.Q.) in order to form a crude emulsion,
and then transferred into a high pressure homogenizer (Avestin).
The emulsification was performed at 9000-40,000 psi while recycling
the emulsion for at least 5 cycles. The resulting system was
transferred into a rotary evaporator, and methylene chloride was
rapidly removed at 40.degree. C., at reduced pressure (30 mm Hg)
for 20-30 minutes. The resulting dispersion was translucent, and
the typical average diameter of the resulting paclitaxel particles
was in the range 50-220 nm (Z-average, Malvern Zetasizer). The
dispersion was further lyophilized for 48 hrs. The resulting cake
was easily reconstituted to the original dispersion by addition of
sterile water or saline. The particle size after reconstitution was
the same as before lyophilization.
[0168] It should be recognized that the amounts, types and
proportions of drug, solvents, proteins used in this example are
not limiting in any way. When compared to toxicity of paclitaxel
dissolved in cremophor formulations, the inventive pharmaceutical
composition containing albumin showed substantially lower
toxicity.
EXAMPLE 49
[0169] This example demonstrates a pharmaceutical composition
comprising albumin and paclitaxel having an intermediate albumin to
paclitaxel ratio.
[0170] Specifically, 135 mg of paclitaxel was dissolved in 3.0 ml
methylene chloride. The solution was added to 27 ml of human serum
albumin solution (5% w/v). Deferoxamine was added as necessary. The
mixture was homogenized for 5 minutes at low RPM (Vitris
homogenizer, model Tempest I.Q.) in order to form a crude emulsion,
and then transferred into a high pressure homogenizer (Avestin).
The emulsification was performed at 9000-40,000 psi while recycling
the emulsion for at least 5 cycles. The resulting system was
transferred into a rotary evaporator, and methylene chloride was
rapidly removed at 40.degree. C., at reduced pressure (30 mm Hg)
for 20-30 minutes. The resulting dispersion was translucent, and
the typical average diameter of the resulting paclitaxel particles
was in the range 50-220 nm (Z-average, Malvern Zetasizer). The
dispersion was further lyophilized for 48 hrs. The resulting cake
was easily reconstituted to the original dispersion by addition of
sterile water or saline. The particle size after reconstitution was
the same as before lyophilization. The calculated ratio (w/w) of
albumin to paclitaxel in this invention composition is
approximately 10.
[0171] It should be recognized that the amounts, types and
proportions of drug, solvents, proteins used in this example are
not limiting in any way. When compared to toxicity of paclitaxel
dissolved in cremophor formulations, the inventive pharmaceutical
composition containing albumin showed substantially lower
toxicity.
EXAMPLE 50
[0172] This example demonstrates the treatment of rheumatoid
arthritis in an animal model with an albumin-paclitaxel
composition.
[0173] The collagen induced arthritis model in the Louvain rat was
used to test the therapeutic effect of albumin-paclitaxel
composition on arthritis. The paw sizes of the experimental animals
were monitored to evaluate the seriousness of arthritis.
[0174] After the arthritis was fully developed (usually .about.9-10
days after collagen injection), the experimental animals were
divided into different groups to receive either albumin-paclitaxel
1 mg/kg q.o.d, or albumin-paclitaxel 0.5 mg/kg+prednisone 0.2 mg/kg
q.o.d. (combination treatment) intraperitoneally for 6 doses, then
one dose per week for three weeks. The paw sizes were measured at
the beginning of treatment (day 0) and every time the drug was
injected. One group received only normal saline as control. By the
end of the experiment, the group receiving albumin-paclitaxel
achieved a 42% reduction of paw size, the combination treatment
group showed a 33% reduction of the paw size, while the control
group had about 20% increase of the paw size relative to the time
when the treatment was initiated.
[0175] In conclusion, the albumin-paclitaxel compositions
demonstrated therapeutic effect on arthritis. The
albumin-paclitaxel combinations are likely to localize at sites of
arthritic lesions by transport through receptor-mediated mechanisms
like gp60.
EXAMPLE 51
[0176] This example demonstrates the use of albumin-paclitaxel
compositions to treat cardiovascular restenosis.
[0177] Paclitaxel eluting stents in animals cause incomplete
healing and, in some instances, a lack of sustained suppression of
neointimal growth in the arteries. The present study tested the
efficacy of a novel systemic delivery albumin-paclitaxel invention
compositions for reducing in-stent restenosis.
[0178] Saline-reconstituted albumin-paclitaxel was tested in 38 New
Zealand White rabbits receiving bilateral iliac artery stents.
Doses of albumin-paclitaxel (1.0 to 5.0 mg/kg paclitaxel dose) were
administered as a 10-minute intra-arterial infusion; control
animals received vehicle (0.9% normal saline).
[0179] In a follow-up chronic experiment, albumin-paclitaxel 5.0
mg/kg was given at stenting with or without an intravenous
3.5-mg/kg repeatalbumin-paclitaxel dose at 28 days; these studies
were terminated at 3 months. At 28 days, mean neointimal thickness
was reduced (p<=0.02) by doses of albumin-paclitaxel>=2.5
mg/kg with evidence of delayed healing. The efficacy of a single
dose of albumin-paclitaxel 5.0 mg/kg, however, was lost by 90 days.
In contrast, a second repeat dose of albumin-paclitaxel 3.5 mg/kg
given 28 days after stenting resulted in sustained suppression of
neointimal thickness at 90 days (p<=0.009 versus single dose
albumin-paclitaxel 5.0 mg/kg and controls) with nearly complete
neointimal healing.
[0180] Although systemic albumin-paclitaxel reduces neointimal
growth at 28 days, a single repeat dose was required for sustained
neointimal suppression. Thus, the inventive composition is suitable
for treatment of cardiovascular diseases such as restenosis.
Inventive compositions comprising pharmaceutical agents other than
paclitaxel, for example rapamycin, other taxanes, epothilones etc,
are all suitable for treatment of restenosis in blood vessels or
artificial blood vessel grafts such as those used for
arterio-venous access in patients requiring hemodialysis.
* * * * *