U.S. patent application number 11/553350 was filed with the patent office on 2007-06-07 for compositions and methods of delivery of pharmacological agents.
This patent application is currently assigned to Abraxis BioScience, Inc.. Invention is credited to Neil P. Desai, Patrick Soon-Shiong, Vuong Trieu.
Application Number | 20070129448 11/553350 |
Document ID | / |
Family ID | 34891317 |
Filed Date | 2007-06-07 |
United States Patent
Application |
20070129448 |
Kind Code |
A1 |
Desai; Neil P. ; et
al. |
June 7, 2007 |
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
serun 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
pharrmaceutical composition, and methods for enhancing transport
and binding of a pharmaceutical agent to a cell.
Inventors: |
Desai; Neil P.; (Los
Angeles, CA) ; Soon-Shiong; Patrick; (Los Angeles,
CA) ; Trieu; Vuong; (Calabasas, CA) |
Correspondence
Address: |
MORRISON & FOERSTER LLP
755 PAGE MILL RD
PALO ALTO
CA
94304-1018
US
|
Assignee: |
Abraxis BioScience, Inc.
Los Angeles
CA
|
Family ID: |
34891317 |
Appl. No.: |
11/553350 |
Filed: |
October 26, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10731224 |
Dec 9, 2003 |
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11553350 |
Oct 26, 2006 |
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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: |
514/776 |
Current CPC
Class: |
A61P 9/00 20180101; Y10S
977/705 20130101; A61K 31/337 20130101; A61K 31/343 20130101; A61K
31/355 20130101; A61K 31/00 20130101; Y10S 977/911 20130101; A61P
35/00 20180101; A61P 37/06 20180101; A61K 9/0078 20130101; A61K
9/19 20130101; Y10S 977/906 20130101; A61K 31/24 20130101; Y10S
977/773 20130101; A61K 31/436 20130101; A61K 38/13 20130101; A61K
9/0019 20130101; A61K 31/4745 20130101; A61K 31/16 20130101; A61P
9/04 20180101; B82Y 5/00 20130101; A61K 9/146 20130101; A61P 31/04
20180101; A61P 39/02 20180101; A61K 31/164 20130101; A61K 31/198
20130101; A61K 31/427 20130101; A61P 19/02 20180101; Y10S 977/779
20130101; A61K 31/05 20130101; A61K 31/7048 20130101; A61P 29/00
20180101; A61K 47/18 20130101; A61P 9/10 20180101; A61K 9/1075
20130101; A61K 47/42 20130101; A61P 43/00 20180101; A61K 9/107
20130101; A61K 31/165 20130101; A61P 23/00 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: |
514/776 |
International
Class: |
A61K 47/00 20060101
A61K047/00 |
Claims
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.
Description
CROSS-REFERENCE TO RELATED PATENT APPLICATIONS
[0001] This patent application is a continuation of patent
application Ser. No. 10/731,224, which claims the benefit of U.S.
Provisional Patent Application No. 60/432,317 filed December 9,
2002, U.S. Provisional Patent Application 60/526,544 filed Dec. 3,
2003, U.S. Provisional Patent Application 60/526,773 filed Dec. 4,
2003, and U.S. Provisional Patent Application 60/527,177 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 fomulated 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 lecthin-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 liotiyronine (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 asetic 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
adminisration-related side effects of drug formulations, alternate
formulations have been attemptec. 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 etics, 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 Clit
Pharmacol., 47(6), 653-60 (1999)). These formulations, however,
induce other side effects (e.g., cardiovascular complications), or
cause destabiliztion of propofol emulsions.
[0006] To overcome the problem of bacterial contamination, propofol
formulations have been developed with antibacteial 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 Intenational 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 periatric 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 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
administation of the pharmaceutical composition into a human, and
wherein the pharmaceutically acceptable carrier comprises
deferoxamine in an amount effective to inhibit oxidation m 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 carrer, wherein the pharmaceutically
acceptable carrier comprises albumin and deteroxamine. 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
pharmacutically acceptable carrier comprises deferoxamine in an
amount effective for inhibiting microbial growth or an amount
effective for inhibiting oxidation in the pharmaceutical
composition.
[0012] The invention also provides a method for enhrncing 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
carier, 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 phamaceutically acceptable
carrier, wherein the pharmaceutically acceptable carrier comprises
albumin in an amount effective to increase tansport 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-pharmceutical 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 carier 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 carrer 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, where 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 carier comprises deferoxamine in an
amount effective to inhibit oxidation in the pharmaceutical
composition.
[0016] Any suiable pharmaceutical agent can be used in the
inventive pharceutical composition. Suitable pharmaceutical agents
include, but are not limted to, anticancer agents or
antineoplastics, antimicrotubule agents, immunosuppressive agents,
anesthetics, hormones, agents for use in cardiovascular disorders,
antiarrythmics, antibiotics, antifungals, antihypertensives,
antiasmatics, analgesics, anti-inflammatory agents, anti-arthriic
agets, 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)),
epotilones, 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 hereof. 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 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 hypersntivity and
fluid retention that can be severe. Epothilone (and derivatives
thereof) also typically is admilnistered in cremophor, and has been
shown to induce severe neutropenia, hypersensivity, 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 admistered
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 +/-02 .mu.M at 22.5.degree. C. (see, e.g., Tonner
et al., Anesthesiology, 77, 926-931 (1992)). As such, propofol is
generally formulaed as an emulsion containing solubmzing 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 at can be used in the composition. The choice
of carier 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 albumn,
inmunoglobulns 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, 65 K 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, JAJM, 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
teatment 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 neural 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 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., Fehiske et al., BiochenL Pharmcol., 30, 687-92
(1981), Vorum, Dan. Med Bull., 46, 379-99 (1999), Kragh-H 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), Purcel et al., Biochim. Biophys. Acta, 1478(1),
61-8 (2000), Altmayer et al., Armneimiateforschung, 45, 1053-6
(1995), and Garrido et al., Rev. Esp. Anestestiol Reanim., 41,
308-12 (1994)). In addition, docetael has been shown to bind to
human plnaproteins (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 phamaceutical compositions results
in a reduction in side effects associated with admistration 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 adminison. 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 lyophilihzion, 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 Streptomyes pilosus, and is capable
of forming iron complexes. Deferoxamine mesylate for injection USP,
for example, is approved by the Food and Drug Administion as an
iron-chelating agent and is available for intramuscular,
subcutaneous, and intravenous adminiration. 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
deferoxamne mesylate is
N-[5-[3-[(5-aminopentyl)hydroxycarbamoyl]-propion-amido]pentyl]-3[[5((N
-hydroxyacetamido)pentyl]-carbamoyl]propionohydroxamic acid
monomethanesulfonate (salt), and its structural formula is
C.sub.2H.sub.48N.sub.6O.sub.8.CH.sub.3SO.sub.3H. As described in
the Examples, deteroxamine, 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 deferoxaiiine, 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 phamacetical 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 prefered
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 for, 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 linited to bile salts of bile acids consisting
of glycocholic acid, cholic acid, chenodeoxycholic acid,
taurocholic acid, glycochenodeoxyrholic acid, taurochenodeoxycholic
acid, litocholic acid, ursodeoxycholic acid, dehydrocholic acid and
others; phospholipids including Lecithin (Egg yolk) based
phospholipids which include the follow phosphtdylcholines:
palmitoyloleoylphosphatidylcholine,
palmitoyllinoleoylphosphatidylcholine,
stearoyllinoleoylphosphatidylcholine
stearoyloleoylphosphatidylcholine,
stearoylaaachidoylphosphatidylcholine, and
dipalmitoylphosphatidylcholine. Other phospholipids including
L-.alpha.-dimyristoylphosphatidylcholine (DMPC),
dioleoylphosphatidylcholine (DOPC), distearyolphosphatidylcholine
(DSPC), hydrogenated soy phosphatidylcholine (HSPC),
D-.alpha.-phosphatidylcholine, .beta.-acetyl-.gamma.-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.-acetyl-.gamma.-O -hexadecyl,
DL-.alpha.-phospatidylcholine, .beta.-acetyl-.gamma.-O-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.acetyl-.gamma.-O-octadecyl,
L-.alpha.-pbospltidylcholine,
.beta.arachidonoyl-.gamma.-O-hexadecyl,
L-.alpha.phosphatidylcholine,
.beta.acetyl-.gamma.-O-(otadec-9-cis-enyl),
D-.alpha.-phosphatidylcholine,
.beta.-arachidonoyl-.gamma.-O-palritoyl, 3-sn-phosphatidylcholine,
2-arachidinoyl-1-stearoyl, L-.alpha.phosphatidylcholine,
.beta.-arachidonoyl-.gamma.-stearoyl, L-.alpha.-
phosphatidylcholine, diarachidoyl, L-.alpha.-phosphatidylcholine,
dibehenoyl, L-.alpha.-phosphatidylcholine,
.beta.-(cis-8,11,14-eicosatrienoyl)-.gamma.-O-hexadecyl,
L-.alpha.-phosphatidylcholine, .beta.-oleoyl-.gamma.-myristoyl,
L-.alpha.-phosphatidylcholine,
.beta.(pyren-1-yl)decanoyl-.gamma.-paiitoyl, 3-sn-phosphatidyl-N,N
-dimethylethanolamine, 1,2-dipalmitoyI,
L-.alpha.-phosphatidyleltntolae, diheptadecanoyl,
3-sn-phosphatidylethanolamine, 1,2dlauroyl,
3-sn-phosphatidylethanolamine, 1,2-dimyristoyl,
3-sn-phosphatidyletoelanae, 1,2dioleoyl,
3-sn-phosphatidylethanolamine, 1,2dipalmitoyl,
L-.alpha.-phosphatidyledmanolamie, dipalmitoyl,
L-.alpha.phosplatidyletlhanolamine, dipalhitoyl, 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)-,2distearoyl-sn
-glycero -phosphoethanolamine sodium (MPEG-DSPE),
L-.alpha.-phosphatidic acid, didecanoyl sodium salt,
L-.alpha.-phosphatidic acid, dihecanoyl sodium salt,
3-sn-phosphatidic acid, 1,2-diymiristoyl 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.-phosphatdyl-DL-glycerol, dipalmitoyl ammonium salt,
L-.alpha.-phosphatidyl-DL-glycerol, distearoyl ammonium salt,
L-.alpha.-phosptidyl-DL-glycerol, .beta.-oleoyl-.gamma.-palmitoyl
ammonium salt, L-.alpha.-phosphatidylinositol ammonium salt,
L-.alpha.-phosphatidylinositol sodium salt,
L-.alpha.-phosphatidyl-L-serme, dioleoyl sodium salt,
L-.alpha.-phosphatidyl-L-serine, and dipalmitoyl sodium salt.
Negatively charged surfts of emulsifiers are also suitable as
additives, e.g., sodium cholesteryl sulfite and the like.
[0025] The phamaeutical 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
prefered 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 acceptle surfactant. The term
"surfactants," as used herein, refers to surface active group(s) of
amphiphile molecules. Surfants can be anionic, cationic, nonionic,
and zwitterionic. Any suitable surfactant can be included in the
inventive pharmaceutical composition. Suitable surfilts include
non-ionic suractants such as phosphatides, polyoxyethylene sorbitan
esters, and tocopheryl polyethylene glycol succinate. Preferable
surfactants are egg lecithin, tween 80, and vitamn 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 surfcnts. Typically, 0.1 to 1.5% by weight of tween 80 or
0.5 to 4% by weight of vie 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 surctants 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, manitol, corn starch, potato starch, microcryslline
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 phamacologically
compatible excipients. Lozenge forms can comprise the active
ingredient in a flavor, usally sucrose and acacia or tragacant, as
well as pastiles 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 igrdient,
such excipients as are known in the art
[0028] Formulations suitable for parenteral administration include
aqueous and non-aqueous, isotonic steile 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,
stabilizs, 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 suspenons 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-xidants, 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 formulatons 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
emulsjfng bases or water-soluble bases. Formulations suitable for
vaginal adinistration 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 pharaceutically 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 contaiing albumin, deferoxamine, a
surfactant, and other water-soluble ingredients, and mixing the oil
with the aqueous phase. Th crude ennikon 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 nanometdrs
(nm). For example, in the case of paclitaxel, docetaxel, rapamyci,
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 adnintson of a pharmaceutical
composition to a human. The method comprises adminiating to a human
a pharmaceutical composition comprising a pharmaceutical agent and
a phmmaceutically acceptable carrier, wherein the pharmaceutically
acceptable camer comprises albumin and deferoxamine. Descrptions 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 anaestesia, 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. intrapulmora administration, oral
administration, inhalation, intravesicular administration, muscular
administration, intra-tracheal administration, subcutaneous
administration, intraocular administration, intrathecal
administration, or transdermal ainistation. For example, the
inventive pharmaceutical composition can be adminiinsted by
inhalation to treat conditions of the respiratory tact. There are
minimal side-effects associated with the inhalation of the
inventive pharmacetnial 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, broncheolits obliterans,
lung cancer, bronchoalveolar carcinoma, and the like.
[0037] The inventive method results in the reduction of one or more
side effects aosociated with administration of a pharmaceutical
composition to a human. Such side effects include, for example,
myelosuppression, neurotoxicity, hypersensitivity, inflammation,
venous irritation, phlebitis, pain, slin iritation, 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 fhiwbiting
microbial growth in a pharmaceutical composition. By "inhibing
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 pharmaceutdcal
composition comprising a phmacutical 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 pacutical composition. In addition, the invention provides a
method for inhibiting oxidation of a pharmaceutical composition.
This method comprises preparing a pharmacutical 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 phalrmaceutical 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 phamaceutical 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 pheuaical 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
granulaton, fluidized-bed drying, and other methods known to those
skilled in the art, deferoxasmne mesylate preferably is applied to
the active pharmaceual agent, and other excipients if presen, as a
solution. The deferoxanine 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
phamaceutical 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 pbancaceutical agent and a
pharmaceutically acceptble carrier, wherein the pharmaceutically
accepable 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 pharmacau composition and
inventive method also are applicable to those same aspects of the
tansport 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
phamaceutical 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 pharmaeutical composition affects the
ability of the pharmaceutical agent to bind and transport the
phareutical 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 phamaceutical 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 pharmaceutical agent will have to be optimized for
different protein and phaceutical agent combinations, generally the
ratio of protein, e.g., albumin, to pharmaeutical 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
preftrnly, 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., Geldeblom et al., supra).
[0042] The pharmaceutical agent can be any suitable pharmaceutical
agent described herein (e.g., propofol, paclitaxel, or doxel). In
addition, the plamaeutical 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 getic material of the cell for a useful
therapeutic effect. The use of protins, such as human serum albumm,
serves as a non-viral veatr 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 inftrmty 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 adinistered 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 stems, 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 endocrre 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 eldothelial cell. In the context of
the inventive method for enhancig 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 transferin, as compared to normal cells. Since tumor
cells are dividing at a rapid rate, they require additional
nutrient sources compared to normal cells. Tumor sies of the
inventive pharmaceutical compositions containig 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 protem
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 tansort 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 gp 16 or gp30, 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
"knockout" 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 usefuil to treat the cancer with
conventional chemotherapy, or with the inventive albumin-drug
phamaceutical compositions (which show high accumulation in
tumors), followed by compounds that block the transport of proteins
to the tumor cell. Blocking compounds can be administed prior to,
or in conjunction with, other chemotherapeutic or anticancer
agents. Thus, any compoumds 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 Sothilone,
albumn-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 tanort 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., inflarnmation (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 fators, 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 cuture with desired cell types
allows for differentiation and proper functioning of islets,
hepaocytes, 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
secete 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 orginatng from other tissues and organs (as descried above)
may also be cocultured with endothelial cells to provide the same
benefit. In addition, the coculture may be ulized to incorporate
genetic material into the target cell tpes. 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 limitng 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. Pats.
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 serm albumin solution (2% w/v). Deferoxamine was added as
necessary. The was homogenized for 5 mmutes 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 emulification was performed at 9000-40,000 psi while recycling
the emulsion for at least 5 cycles. The resulting system was
transfenred into a rotary evaporator, and methylene chloride was
rapidly removed at 400.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, Malvem Zetasizer). The
dispersion was further lyophilzed for 48 hrs. The resulting cake
could be easily reconstituted to the original dispersion by
addition of sterile water or saline. The particle size aeter
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 substantay lower
toxicity.
EXAMPLE 2
[0053] This example demonstrates the preparation of a phmaceutical
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 homogenied for 5 minutes at
low RPM (Vitis homogenizer, model Tempest I.Q.) in order to form a
crude emulsion, and then transferred into a high pressure
homogenizer (Avestin). The emulisification was performed at
9000-40,000 psi while reycling the emulsion for at least 5 cycles.
The resulting system was tansferred 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 hr. The
resulting cake was easily reconsti to the originad dispersion by
addition of sterile water or saline. The particle size after
reconstitution was the same as before lyophilization.
[0054] It should be recognzed 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 concentraion 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 homogeneer (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 resuling dispersion was translucent and the typical average
diameter of the resuling 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 sterle
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 epoilone 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 tansfered
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 orginal
dispersion by addition of sterile water or saline. The particle
size after reconstitution was the same as before lyophilization. It
should be recognzed 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.
EXAMLE 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). Deferoxarnne 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 traslucent and the typical average
diameter of the resulting paricles was in the range 50-220 nm
(Z-average, Malvem Zetasizer). The dispersion was further
lyophilized for 48 hours. The resulting cake was easily
reconstituted to the original dispersion by addition of steile
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 preparaton of pharmaceutical
compositions comprising docetaxel and albumin. 30 mg of docetaxel
was dissolved in 2 ml chloroforn/ethanol. The solution was then
added into 27.0 ml of human serum albumin solution (3% w/v).
Deferoxamine was added as necessary. The mix was homogenized fr 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
homogenlzer (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 resuing dspersion was translsucent 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 reconsttution 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 phamacetical composition comprising albumin
showed substantially lower txicity.
EXAMPLE 8
[0060] This example demonstrats 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
(Vitis homogen model Tempest I.Q.) in order to form a crude
emulsion, and then transferred into a high pressure homogenizer
(Avestn). 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 resuling 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
reconstiteeed to the original dispersion by addition of sterile
water or saline. The particle size after reconstituton was the same
as before lyophiiiaaon. 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 substaintially lower toxicity.
EXAMPLE 9
[0061] This example demonstrates the preparation of pharmaceucal
compositions comprising a taxane IDN5390 and albumin. 150 mg of
IDN5390 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 solvens and oils were used and these compositions are also
contemplated as part of the invention. A small quantity of a
negatively ciarged component was also optionally added, e.g,
benzoic acid (0.0010%-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 tnsferred into a high pressure homogenizer
(Avestin). The emulsification was performed at 9000-40,000 psi
while recycling the emidsion for at least 5cycles. 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 albun. 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 optionaly added, e.g.,
benzoic acid (0.001%.5%) The solution was then added into 27.0 ml
of human serum albumin solution (5% w/v). Deftioxamine 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 a least 5 cycles. The resulting
system was transferred into a Rotavap and solvend was rapidly
removed at 40.degree. C., at reduced pressure (30 mm Hg) for 20-30
minuts. The resulting dispersion was translucent and the typical
average diameter of the resiuting particles was in the range 50-220
nm (Z-average, Malvern ). The dispersion was e lyphilized for 48
hours. The resulting cake was easily reconstited to the orginal
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 limitng.
When compared to toxicity of the IDN5109 dissolved in tween, the
phawiaceutcal 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 albunin 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 tansferred 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 liiting im 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 minues 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 emulsificion was perfomed at 9000-40,000
psi while recycling the emulsion for at least 5 cycles. The
resulting system was sferred into a Rotavap and methylene chloride
was rapidly removed at 400.degree. C., at reduced pressure (30 mm
Hg) for 20-30 miutes. 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 lyophilzation.
EXAMPLE 13
[0065] This example demonstrates the preparation of a
pharraceutical 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 (Vitis homogenizer, model Tempest I.Q.) in order to form a
crude emulsion, and then transfeered 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 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 liming 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 (Avestn). The
emulsification was performed at 9000-40,000 psi while recycling the
emusion 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 transuent, and the typical average
diameter of the resulting amphoterin particles was between 50-220
nm (Z-average, Malven Ztasizer). The dispersion was further
lyophilized for 48 hrs. The resulting cake could be easly
reconstitd 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 pharmacoknetics 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
albumnin-paclitaxel pharmaceutical compositions over
cremophor-paclitaxel (Taxol) pharmacutical 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.mas, suggesting more rapid
distribution of alburmin-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 be 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
pharmaodynamics, i.e., enhanced inta-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.mas 1.2-fold higher, and tumor
half-life 1.7-fold longer than for Taxol in tumor beaing mice.
EXAMPLE 16
[0069] This example demonrtrates 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
phamaceutical 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 albumm in a pharmaceutical composition
comprising paclitaxel results in a substantial reduction in side
effects and toxicity when compared to conventional pharmceutical
compositions comprising paclitaxel.
EXAMPLE 17
[0071] This example demonstrates the clinical effects of a
prmaceutal composition comprising paclitaxel and albumin in
humans.
[0072] Clinical studies in over 500 humnan patients to date provide
evidence supporting the reduction in toxicity and side-effects for
a phamaceutical composition comprising paclitaxel and albumin
("albumm-paclitaxel") when compared with cremophor-paclitaxel
compositions (Taxol). In a phase I study of 19 patients, the
maximuum 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
iritation, 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
admiisterd 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/m2 every 3 weeks in 43 and 63 patients
restively, 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/m2 respectively. Severe neuropathy occurred in 0% and 14% of
patients for 175 mg/m.sup.2 and 300 mg/m.sup.2 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 han
eperienced 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 pharmaceutial composition comprising albumin and
paclitaxei.
[0077] An in vitro cytotoxicity study comparing the effect of
albmiin-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 ovaran, 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 (<05) in all
tumors except the lung tumor (p=5).
[0079] In the MX1 mammrny 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 caner
(N=31) and cancer of the anal canal (N=12). The dose escalated from
120-300 mgmm.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=11), while
patients with cancer of the anal canal exhibited a response rate
64% (N=11).
EXAMPLE 20
[0082] This example demonses 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.mu.m 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 urrtu dissolved.
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 15 cycles at
5.degree. C. Alterately, discrete passes through the homogenizer
were used. The inal emulsion was filtered (02 .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 injecon 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 phamacetcal
composition containing 5% oil and comprising propofol and
albumin.
[0085] An oil-in-water emulsion containing 1% (y 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.mu.m 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 10,000 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%; 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.,
deferxamine (0.001-0.1%), were optionally added.
EXAMPLE 22
[0086] This example demonses 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 prepare 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 untll 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 homogized 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
pamaceutical 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
225%; water for injection q.s. to 100; pH 5-8.
EXAMPLE 23
[0088] This example demonstes the preparation of a phamnaceutical
composition comprising propofol, albunn, and vitamin E-TGS, which
is free of oil.
[0089] Using the procedure simlar 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 albumnin (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.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.
Alterately, 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 containig 1% (y 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 trough 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 homogeniner 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 albumi 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 sired 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%-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 sied until dissolved. The
aqueous phase was passed through a filter (0.21 .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 homog 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.,
phospbatidyl 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 homogenizd 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.
[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 deteried as follows.
Solubility of propofol was tested in water and in solutions
containing albumin. 2 .mu.L of propofol was added to 10 mL of a
water or albumin solution and stirred for 2 hours in a scintillion
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.
Solubifity of propofol in albumin solutions was dependent on the
concentration of albumin and increased to 0.44 mg/ml when the
albumim 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, indicaing a sustanial binding of propofol with
albumin. Based on these results, addition of albumin to
pharmaceutical compositions comprising propofol result in a
decrease in the amoxmt 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 ultrfiltration 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 contaiing 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 corrspond
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 administrtion of a pharmaceutical
composition comprising propofol and albumin to humans.
[0107] A randomized, doubleblind clinical trial was conducted to
compare adverse skin sensations of a pharmaceutical composition
comprsing 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. Adult human subjects of either sex were
eligible for participation if they had unbroken, apparently normal
skin of dorsal side of their hands.
[0108] The formulations originally stored in a refrigerator were
brought to room temperature and then 10 .mu.L of the formutons 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. TABLE-US-00001 TABLE 1 % of subjects % of
subjects with ABI-Propofol with Diprivan sensation sensation Mild
warm Mild warm Order of a test or stinging or stinging on a subject
or biting No sensation or biting No sensation 1st incidence 0.0
100.0 75 25
EXAMPLE 31
[0109] This example demonstrates the use of deferoxamine as
antioxidant in a pharmaceutical composition comprising
propofol.
[0110] 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. TABLE-US-00002 TABLE 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%
[0111] TABLE-US-00003 TABLE 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%
[0112] 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
[0113] This example demonstrates intrapulmonary delivery of a
pharmaceutical composition comprising paclitaxel and albumnm
(ABI-007).
[0114] The purpose of this study was to determine the time course
of [.sup.3H]ABI-007 in blood and select tissuces following
intratracheal instillation to Sprague Dawley rats.
[0115] The target volume of the intratacheal dose formulation to be
admiistered 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.
[0116] Blood samples of approximately 250 .mu.L were collected from
the indwelling jugular cannulas of JVC rats at the following
predetmined 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 2h, were collected via cardiac punctre from
anesthetized rats at sacifice. 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.
[0117] For all rats, the maximum concentration of tritium in blood
was observed at 5 min (0.0833 hr) post dosing. The elimination
haf-life of tritium, detemined over the time interval ftom 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
exeriments are set forth below in Table 4. TABLE-US-00004 TABLE 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 t1/2beta (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
[0118] The mean blood concentration of [.sup.3H]ABI-007 -derived
radioactivity after an itravenous dose to rats was analyzed as a
function of time in order to evaluate the bioavailabilty of tritium
derived from an intraacheal 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.
[0119] 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, respecfively. The average apparent clearance of
tritium from blood was 0.1235+/-0.0180 L/hr (see Table 4
above).
[0120] Tritium derived from [.sup.3H]ABI007 was absorbed and
distributed after intratracheal administaton. 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 1 min after the intratracheal dose. A
maximum of less than 1% of the dose was recovered in other tissues,
excluding the gastroitestinal tract, at all time points
examined.
[0121] Based on results from a previously conducted intravenous
dose study with [.sup.3]Capxol.TM., the bioavailability of tritium
derived from the intatracheal 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 usig invention compositions with albumin was rapidly
bioavailable indicating excellent transport across pulmonary
endothelium. No toxicity in the animals was noted, wbich was
surpsing since pulmonary delivery of cytotoxics is known to cause
lung toxicities.
[0122] A fair amount of radioactivity was present in the
gastrointestial 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 excetion or clearance
of tritium from the respiratory tract via mucociliary clearance
with subsequent swallowing.
EXAMPLE 33
[0123] This example demonstrates investigation of Aerotech II and
Pari nebulizers for pulmonary delivery of pharmaceutical
compositions comprsig paclitaxel and albumin.
[0124] 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/in.sup.2 for Pari), run time (15 to 60 seconds), sample volume
(1.5 mL), ABI-007 pacliael concentration (5, 10, 15 and 20
mg/mL).
[0125] 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.
[0126] 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 nebultizers.
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.
[0127] In conclusion, the nanoparticle/albumin formulation of
paclitaxel (ABI-007) shows excellent bioavailility 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.
EXAMPIE 34
[0128] 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 insiluaton to Sprague
Dawley rats as compared to intravenous installation.
[0129] 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 adinishaton 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.
[0130] 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, and45 minutes (min) and 1, 4, 8, and 24
hours (h). All blood samples analyzed were dispensed into
pre-weighed sample tues, and the sample tubes were reweighed, and
the weight of each sample was calculted by subtraction. The blood
samples collected were assayed for total rapamycin concentration
using LC/MS/MS.
[0131] 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 pulnonary route using a pharmaceutical composition
comprising albumin was calculated to be 109%, indicating excellent
transport across pulmonary endothelium.
EXAMPLE 35
[0132] 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 putnonary absorption of rapamycin
in tissue following intratracheal installation to Sprague Dawley
rats as compared to intravenous installaion.
[0133] 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.,
Philadelplaa, Pa.; purchased from DeLong Distributors, Long Branch,
N.J.) attached to a 1-mL gas-tight, luer-lock syringe. The
appropriate volume of dose preparaton was drawn into the dosing
apparatus, the filled apparatus was weighed and the
weight-recorded. A catheter was placed in the trachea of the
anestheied animal, the microsprayer portion of the dosing apparatus
was placed into the trachea through the catheter, and the dose was
administered. After dose admanstration 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.
[0134] 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 wee collected and analyzed for
total rapamycin concenfaton using LC/MS/MS. The results indicate
that rapamycm conoentration 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, pulnonary delivery of rapamycin may be suitable for
the treatment of a condition (i.e., lung plantation), wherein high
local concention of rapamycin would be beneficial.
[0135] This example demonruates oral delivery of a pharmaceutical
composition comprising paclitaxel and albumin (ABI700).
[0136] Tritated ABI-007 was utilized to deterniine 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 preteated
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 detennined by comparison with intavenous
data previously obtained. The results are set forth below in Table
5. TABLE-US-00005 TABLE 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
[0137] 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).
[0138] An oral bioavailability of 44% was seen for ABI-007 alone.
This is dramatically higher than is seen for other formulations of
pacitaxel. The bioavailability inreased to 121% when animals were
treated with cyclosporine (CsA). This is expected as CsA is a known
suppressor of the p-glycoprotin 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 binary excretion of pacitaxel into the GI tract. Other
known suppressors or enhancers of absorption may be also utilized
for this purpose.
EXAMPLE 37
[0139] This example demonstrates improved penetration of paclitaxel
into red blood cells and tumor cells upon admiisttion of a
pharmaceutical composition comprising paclitaxel and albumn.
[0140] Human MX-1 breast tumor fragments were implanted
subcutaneously in athymic mice. A pharmaceutical composition
comprising paclitaxel and albulin ("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. Plasm, 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 admiistaion. Tumor pharmacokinetic (AUC and absorption
constant) was analyzed using WinNonlin, Pharsight, USA.
[0141] Paclitaxel-albumin exhibited rapid partitioning into red
blood cells (RBCs) as shown by a rapid drop of the plasma/blood
radioactivity ratio to unty 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.
[0142] Paclitaxel-albumin exhibited a rapid partitioning into tumor
tissue with an absorption constant (K.sub.a) that was 3.3X 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
[0143] This example demonstrates the safety of a pharmaceutical
composition comprising paclitaxel and albumin administed to
mice.
[0144] Athymnic 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 deatis 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
[0145] This example demonstrates a novel paclitaxel transport
mechanism across microvessel endothelial celis (EC) for
paclitaxel-albumin compositions.
[0146] Nanopartcles and albumin-paclitaxel compositions can
accumulate in tumor tissue due to EPR effect resulting from `leaky`
vessels in a tumor. An albumin seic gp60 receptor (albondin)
transported albumin across EC by transcytosis of the receptors
within caveolae at the cell surfice. 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
[0147] This example demonstrates an increase in endothelial
transcytosis of pharmaceutical compositions comprising paclitaxel
and albumin as compared to Taxol.
[0148] Human lung microvessel endothelal cells (HLMVEC) were grown
to confluence on a transwell. The inventive pharmaceutical
composition comprising paclitaxel and albumin, or Taxol containig
fluorescent paclitaxel (Flutax) at a concentration of 20 .mu.g/mL,
was added to the upper transwell chamber.
[0149] The tansport 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 HMVEC monolayer. Transport of
paclitaxel from the albumin-paclitaxel composition was much faser
than paclitaxel from Taxol in the presence of 5% HSA (physiological
concentration). Transport rate contstant K.sub.t for the
albumin-paclitaxel composition and Taxol were 1.396 hr.sup.-1 and
0.03 hr.sup.-1, respevely. The total amount of paclitaxel
transported across the monolayer was three times higher for the
albumin-paclitaxel composition than Taxol.
EXAMPLE 41
[0150] This example demofltrates improved endothelial cell (EC)
binding by pharmaceutical compositions comprising paclitaxel and
albumin as compared to Taxol.
[0151] 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 reacd to the HUVEC at various final
concentrations. Biding of paclitaxel to cells was inhibited by
Cremophor. Inhibition was exhibited by an IC.sub.50 of 0.02% of
Creinophor 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, litle or no binning was observed for Taxol.
EXAMPLE 42
[0152] This example demonstrates improved albumin binding by
phamaceutical compositions comprising paclitaxel and albumin as
compared to Taxol.
[0153] Human Serum Albumiin (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. hihbition 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
[0154] This example demonstrates increased transfer of paclitaxel
to albumin for pharmaceutical compositions comprising paclitaxel
and alburmin as compared to Taxol.
[0155] 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 scaningn
fluorometer. The transfer of paclitaxel to HSA was more rapid for
the albumin-paclitaxel composition versus Taxol. More paclitaxel
co-electeophorsed 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 tansferred 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 imporatant process in vivo.
EXAMPLE 44
[0156] This example demonstrates that the glycoprotein receptor
gp60 is responsible for binding and transcytosis of
albumin-paclitaxel.
[0157] Fluorescent labeled paclitaxel (Flutax) albulmin
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
[0158] This example demonstrates that increasing amounts of albumin
can compete with binding of paclitaxel.
[0159] Albumin was immobiliz 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 immobiize 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
[0160] This example deinonstrates that lower amounts of albumin in
the inventive pharmaceutical composition results in stable
compositions.
[0161] To investigate if lower amounts of albumin in compositions
would affect stability of the inventive pharmaceutical composition,
albumin-paclitaxel compositions with low amounts of albunin 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
[0162] This example demostrates a pharmaceical composition
comprising albumin and paclitaxel having a high albumin to
paclitaxel ratio.
[0163] 30 mg of paclitaxel was dissolved in 3.0 ml methylene
chloride. The solution was added to 27.0 ml of humnan 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 sferred
into a high pressure homogenzer (Avestin). The emulsification was
performed at 9000-40,000 psi while recycling the emulsion for at
least 5 cycles. The resulting system was transfee 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 tanslucent, and the typical average
diameter of the resulting paclitaxel particles was in the range
50-220 nm (Z-average, Malvern Zetasizer). The dispersion was fbrter
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
lyophilizaton.
[0164] It should be recognd 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
[0165] This example demonstrates a pharmaceutical composition
comprising albumin and paclitaxel having a low albumin to
paclitaxel ratio.
[0166] 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 transfured 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 orgimil dispersion by addition of
sterile water or saline. The particle size after reconstitution was
the same as before lyophiliation.
[0167] 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 siustantially lower
toxicity.
EXAMPLE 49
[0168] This example demonstrates a pharmaceutical composition
comprising albumin and paclitaxel having an intermediate albumin to
paclitaxel ratio.
[0169] 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 fiurter lyophilized for 48 hrs. The resulting cake
was easily reconstitrted to the orginal 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.
[0170] 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 contining albumin showed substantially lower
toxicity.
EXAMPLE 50
[0171] This example demonstrates the treatment of rheumatoid
ardaritis in an animal model with an albumin-paclitaxel
composition.
[0172] 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.
[0173] 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 0.5mg/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 measue at the
beginning of treatment (day 0) and every time the drug was
injected. One group receved only normal saline as control. By the
end of the expeiment, the group receiving albunin-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.
[0174] 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
[0175] This example demonstrates the use of albumin-paclitaxel
compositions to treat cardiovascular restenosis.
[0176] 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.
[0177] 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 palitaxel dose) were
administered as a 10 minute intra-arterial infusion; control
animals received vehicle (0.9% normal saline).
[0178] In a follow-up chronic experiment, albumin-paclitaxel 5.0
mg/kg was given at stening with or without an intravenous 3.5-mg/kg
repeat albumin-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 suppression of neointimal
thickness at 90 days (p<=009 versus single dose
albumin-paclitaxel 5.0 mg/kg and controls) with nearly complete
neointimal healing.
[0179] Although systemic albumin-paclitaxel reduces neointimal
growth at 28 days, a single repeat dose was required for sustained
neointl 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.
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