U.S. patent application number 17/194980 was filed with the patent office on 2021-12-09 for injectable melphalan compositions comprising a cyclodextrin derivative and methods of making and using the same.
The applicant listed for this patent is Cydex Pharmaceuticals, Inc.. Invention is credited to Stephen G. Machatha, James D. Pipkin.
Application Number | 20210379000 17/194980 |
Document ID | / |
Family ID | 1000005770037 |
Filed Date | 2021-12-09 |
United States Patent
Application |
20210379000 |
Kind Code |
A1 |
Pipkin; James D. ; et
al. |
December 9, 2021 |
INJECTABLE MELPHALAN COMPOSITIONS COMPRISING A CYCLODEXTRIN
DERIVATIVE AND METHODS OF MAKING AND USING THE SAME
Abstract
The present invention is directed to pharmaceutical compositions
comprising melphalan and a cyclodextrin derivative, and methods of
making and using the same.
Inventors: |
Pipkin; James D.; (Lawrence,
KS) ; Machatha; Stephen G.; (Overland Park,
KS) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Cydex Pharmaceuticals, Inc. |
Lenexa |
KS |
US |
|
|
Family ID: |
1000005770037 |
Appl. No.: |
17/194980 |
Filed: |
March 8, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14108169 |
Dec 16, 2013 |
10940128 |
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17194980 |
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12790724 |
May 28, 2010 |
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14108169 |
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61182560 |
May 29, 2009 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/6951 20170801;
A61K 9/0019 20130101; A61K 31/198 20130101; C08B 37/0015 20130101;
C08L 5/16 20130101; B82Y 5/00 20130101; A61K 47/40 20130101 |
International
Class: |
A61K 31/198 20060101
A61K031/198; A61K 9/00 20060101 A61K009/00; B82Y 5/00 20060101
B82Y005/00; A61K 47/69 20060101 A61K047/69; C08B 37/16 20060101
C08B037/16; C08L 5/16 20060101 C08L005/16; A61K 47/40 20060101
A61K047/40 |
Claims
1. A method of treating a subject suffering from a neoplastic
disorder, the method comprising: diluting a composition with an
aqueous diluent to provide a dilute pharmaceutical composition
comprising 25 mg to 125 mg of melphalan and a cyclodextrin
derivative of formula 1: ##STR00022## wherein n is 4, 5 or 6;
wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 are independently --H, a
straight-chain or branched C.sub.1-C.sub.1-(alkylene)-SO.sub.3--
group, or an optionally substituted straight-chain or branched
C.sub.1-C.sub.6 group; wherein at least one of R.sub.1, R.sub.2,
P.sub.3, P.sub.4, R.sub.5, R.sub.6, R.sub.7, K.sub.8 and R.sub.9 is
a straight-chain or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3
group; wherein the dilute pharmaceutical composition has, a pH of
about 4 to about 6; wherein the cyclodextrin derivative is present
in a concentration of at least 50:1 (w/w) relative to: the
melphalan, wherein the melphalan in the dilute, pharmaceutical
composition degrades by 2% or less at about 25Q C. within 0.5 hours
after the diluting; and administering the dilute pharmaceutical
composition, by injection to the subject in need thereof.
2. The method of claim 1, wherein the neoplastic disorder is
selected from: myeloma, Multiple myeloma, melanoma, acute
myelogenous leukemia, malignant melanoma, breast cancer, ovarian
cancer, testicular cancer, advanced prostate cancer, a
neuroendocrine cancer, metastatic melanoma, a metastatic
neuroendocrine tumor, a metastatic adenocarcinoma tumor,
hepatocellular carcinoma, osteogenic sarcoma, polycythemia
veraplasma; plasma cell neoplasm, amyloidosis, scleromyxedema, and
combinations thereof.
3. The method of claim 2, wherein the neoplastic disorder is
multiple myeloma and the administering is systemic and provides,
palliative treatment of the multiple myeloma.
4. The method of claim 1, wherein at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9.
is a hydroxy-substituted-C.sub.3 group.
5. The method of claim 1, wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3-- group having a degree of
substitution of 4 to 8 per cyclodextrin derivative, and the
remaining substituents are --H.
6. The method of claim 1, wherein at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is
substituted with a straight-chain C.sub.4-(alkylene)-SO.sub.3--
group.
7. The method of claim 1, wherein the cyclodextrin derivative is a
compound of formula IL ##STR00023## wherein R.dbd.(H).sub.21-x, or
((CH.sub.2).sub.4SO.sub.3Na.sup.+).sub.x and x=6.0-7.1; wherein the
pharmaceutical composition comprises about 50 mg of melphalan as a
hydrochloride salt; and wherein the cyclodextrin derivative is:
present in a concentration, of 50:1 to 100:1 (w/w) relative to the
melphalan.
8. The method of claim 1, wherein, the dilute pharmaceutical
composition is substantially free of an alcohol.
9. The method of claim 1, wherein the aqueous diluent is a saline
solution.
10. The method of claim 1, wherein the, subject suffering from the
neoplastic disorder is pediatric subject.
11. The method of claim 1, wherein the melphalan in the dilute
pharmaceutical composition degrades by 4% or less at 25.degree. C.
within 10 hours after the diluting:
12. The method of claim 1, wherein the dilute pharmaceutical
composition is stored about 0.5 hours to about 18 hours prior to
the administering.
13. The method of claim 1, wherein the administering, provides a
melphalan C.sub.max in the subject suffering from a neoplastic
disorder that is at least 20% or greater than a melphalan C.sub.max
provided by a melphalan formulation containing an equivalent dose
of melphalan and lacking the cyclodextrin derivative.
14. The method of claim 1, wherein the administering provides a
melphalan AUC.sub.0-t in the subject suffering from a neoplastic
disorder that is at least 20% or greater than a melphalan
AUC.sub.0-t provided by a melphalan formulation containing an:
equivalent dose of melphalan and lacking the cyclodextrin
derivative.
15. A method for conditioning a subject for whom a stem cell
transplantation has been indicated, the method comprising:
administering a melphalan dose of 50 mg/m.sup.2 to 300 mg/m.sup.2
per day to the subject for whom a stem cell transplantation has
been indicated, wherein the melphalan dose is administered in a
pharmaceutical: composition comprising melphalan and a cyclodextrin
derivative of formula I: ##STR00024## wherein n is 4, 5 or 6;
wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 are independently --H, a
straight-chain or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3--
group, or an optionally substituted straight-chain or branched
C.sub.1-C.sub.6 group; wherein at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is
a straight-chain or branched C1-C8-(alkylene)-SO.sub.3 group;
wherein the pharmaceutical composition has a pH of about 4 to about
6; and wherein the cyclodextrin derivative is present in a ratio of
at least 25:1 (w/w) relative to the melphalan.
16. The method of claim 15, wherein the administering is for a
period of two or more days.
17. The method of claim 15, wherein, the subject in need of the
stem cell transplantation is a pediatric subject.
18. The method of claim 15, wherein the administering is performed
intravenously.
19. The method of claim 15, wherein, the administering is performed
via a limb perfusion.
20. The method of claim 15, wherein at least one of R.sub.1,
R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and
R.sub.9 is a hydroxy-substituted-C.sub.3 group.
21. The method of claim 15, wherein the cyclodextrin derivative is
a compound of formula II: ##STR00025## wherein R.dbd.(H).sub.21-x
or (--(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1;
wherein the pharmaceutical composition comprises about 200 mg of
melphalan as a hydrochloride salt; and wherein the cyclodextrin
derivative is present in a ratio of 25:1. to 35:1 (w/w) relative to
the: melphalan.
22. The method of claim 15, wherein the subject for whom. A stem
cell transplantation has: been indicated suffers from a disease or
disorder selected from: myeloma, multiple myeloma, a lymphoma,
non-Hodgkin lymphoma, leukemia, acute myeloid leukemia, Hodgkin's
disease, acute lymphoblastic leukemia, a: myelodysplastic syndrome,
a myeloproliferative disorder; chronic myelogenous leukemia,
neuroblastoma, aplastic anemia, chronic granulocytic leukemia, a
neuroblastoma, sickle-cell disease, osteogenic sarcoma, Ewing's
sarcoma, a desmoplastic small round cell tumor, plasma cell
neoplasm, amyloidosis, scleromyxedema, and combinations
thereof.
23. The method of claim 15, comprising diluting a concentrated
melphalan composition with an aqueous diluent to provide the
pharmaceutical composition,
24. The method of claim 23, wherein the concentrated melphalan
composition comprises 50 mg to 500 mg of melphalan.
25. The method of claim 23, wherein the concentrated melphalan
composition comprises about 200 mg.
26. The method of claim 15, wherein the pharmaceutical composition
is substantially free of an alcohol.
27. The method of claim 23, wherein the aqueous diluent is a saline
solution.
28. The method of claim 23, wherein the melphalan in the
pharmaceutical composition degrades by 4% or less at about
25.degree. C. within 10 hours after the diluting.
29. The method of claim 23, wherein the pharmaceutical composition
is stored about 0.5 hours to about 12 hours prior to the
administering.
30. The method of claim 15, wherein the administering provides a
melphalan C.sub.max in the subject for whom a stem cell
transplantation has been indicated that is at least 20% or greater
than a melphalan C.sub.max provided by a melphalan formulation
containing an equivalent dose of melphalan and lacking the
cyclodextrin derivative.
31. The method of claim 15, wherein the administering provides a
melphalan AUC.sub.0-t in the subject for whom a stem cell
transplantation has: been indicated that is at least 20% or greater
than a melphalan. AUC.sub.0-t provided by a. melphalan formulation
containing an equivalent dose of melphalan and lacking the
cyclodextrin derivative.
32. A pharmaceutical composition comprising: 25 mg to 125 mg of
melphalan as a hydrochloride salt; an optional buffer; and a
cyclodextrin derivative of formula 1: ##STR00026## wherein n s 4, 5
or 6; wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 are independently --H, a
straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3.sup.- group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R1, R2, R3, R4, R5, R6, R7, R8 and R9 is a,
straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3.sup.- group; wherein the
pharmaceutical composition has a pH of about 4 to about 6, wherein
dilution of the pharmaceutical composition with an aqueous solution
provides a dilute pharmaceutical composition in which the melphalan
degrades by 2% or less at about 25.degree. C. within 5 hours after
the dilution; and wherein the cyclodextrin derivative is present in
a ratio of 50:1 to 100:1 (w/w) relative to the melphalan.
33. The pharmaceutical composition of claim 32, wherein at least
one of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 is a hydroxy-substituted-C.sub.3
group.
34. The pharmaceutical composition of claim 32, wherein the
cyclodextrin derivative is a compound of formula: II: ##STR00027##
wherein R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4SO.sub.3Na.sup.+).sub.x, and x=6.0-7.1; wherein
the pharmaceutical composition comprises about 50 mg of melphalan
as a hydrochloride salt; and wherein the cyclodextrin derivative,
is present in a ratio, of about 55:1 (w/w) relative to the
melphalan.
35. A pharmaceutical composition comprising; 150 mg to 250 mg of
melphalan as a hydrochloride salt; an optional buffer; and a
cyclodextrin derivative of formula I: ##STR00028## wherein n is 4,
5 or 6; wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5,
R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are independently --H, a
straight-chain or branched C.sub.1-C.sub.8(alkylene)-SO.sub.3--
group, or an optionally substituted straight-chain or branched
C.sub.1-C.sub.6 group; wherein at least one of R1, R2, R3, R4, R5,
R6, R7, R8 and R9 is a straight-chain or branched
C.sub.1-C.sub.8(alkylene)-S0.sub.3.sup.- group; wherein the
pharmaceutical composition has a pH of about 4 to about 6, wherein
dilution of the pharmaceutical composition with an aqueous solution
provides a melphalan solution in. which the melphalan degrades by
2% or loss a about 25.degree. C. within 5 hours after the dilution;
and wherein the cyclodextrin derivative is present in a ratio of
25:1 to 35:1 (w/w) relative to the melphalan.
36. The pharmaceutical composition of claim 35, wherein at least
one of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 is a hydroxy-substituted C.sub.3
group.
37. The pharmaceutical composition of claim 35, wherein the
cyclodextrin derivative is a compound of formula II: ##STR00029##
wherein R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4SO.sub.3Na.sup.+).sub.x, and x=6.0-7.1; wherein
the pharmaceutical composition comprises about 200 mg of melphalan
as a hydrochloride salt; and wherein the cyclodextrin derivative is
present in a ratio of about 30:1 (w/w) relative to the
melphalan.
38. A pharmaceutical kit comprising: a first container comprising
25 mg to 125 mg of melphalan as a hydrochloride salt and an
optional water-soluble polymer; and a second container comprising
an aqueous diluent, an optional buffer, and a cyclodextrin
derivative of formula I: ##STR00030## wherein n is 4, 5 or 6;
wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 are independently --H, a
straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-S0.sub.3.sup.- group, or an optionally
substituted straight-chain or branched C1-C6 group; wherein at
least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 is a straight-chain or branched
C.sub.1-C.sub.8(alkylene)-S0.sub.3.sup.- group; wherein the
cyclodextrin derivative is present in the second container in a
concentration of at least 50:1 (w/w) relative to the melphalan; and
wherein combining the first container and. the second. container
provides a dilute pharmaceutical composition having a pH of about 4
to about 6 that degrades by 2% or less at about 25.degree. C.
within 5 hours after the diluting.
39. The pharmaceutical kit of claim 38, wherein the first container
comprises povidone in an amount of 10 mg to 3.0 mg, and the second
container comprises a pH-adjusting agent in a concentration
sufficient to provide a pH of about 4 to about 6 when the first
container and the second container are combined.
40. A pharmaceutical kit comprising; a first container comprising
150 mg to 250 mg of melphalan as a hydrochloride salt and. an
optional water-soluble polymer; and a second container comprising
an aqueous diluent, an optional buffer, and a cyclodextrin
derivative of formula I: ##STR00031## wherein n is 4, 5 or 6;
wherein R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 are independently --H, a
straight-chain or branched C.sub.1-C.sub.8(alkylene)-S0.sub.3
group, or an optionally substituted straight-chain or branched
C.sub.1-C.sub.6 group; wherein at least one of RI, R2, R3, R4, R5,
R6, R7, R8 and R9 is a straight-chain or branched
C.sub.1-C.sub.8(alkylene)-S0.sub.3.sup.- group; wherein the
cyclodextrin derivative is present in the second container in a
concentration of 25:1 to 35:1 (w/w) relative to the melphalan; and
wherein combining the first container and the second container
provides a dilute pharmaceutical composition having a pH of about 4
to about 6 that degrades by 2% or less at about 25.degree. C.
within 5 hours after the diluting.
41. The pharmaceutical kit of claim 40, wherein the first container
comprises povidone in an amount of 10 mg to 30 mg, and the second
container comprises a pH-adjusting agent in a concentration
sufficient to provide a pH of about 4 to about 6 when the first
container and the second container are combined.
42. The pharmaceutical kits of claim 40, wherein the cyclodextrin
derivative is a compound of formula II: ##STR00032## wherein
R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4SO.sub.3.sup.-Na.sup.+).sub.x, and x=6.0-7.1;
wherein the first container comprises about 200 mg of melphalan as
a hydrochloride salt; and wherein the cyclodextrin derivative is
present, in the second container in an amount of about 30:1 (w/w)
relative to the melphalan.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 14/108,169, filed Dec. 16, 2013, which is a
continuation of U.S. patent application Ser. No. 12/790,724, fled
May 28, 2010, which claims the benefit U.S. Appl. No. 61/182,560,
filed May 29, 2009, each of which is incorporated herein by
reference in its entirety.
BACKGROUND OF THE INVENTION
Field: of the Invention
[0002] The present invention relates to pharmaceutical compositions
comprising melphalan and a cyclodextrin derivative and methods of
making and using the same, for example, to treat disorders and
diseases that are therapeutically responsive to melphalan.
Background of the Invention
[0003] Melphalan is an alkylating agent of the bischloroethylamine
type, and is active against both resting and rapidly dividing tumor
cells. See, e.g., U.S. Pat. Nos. 3,032,584, 3,032,585, and
4,997,651. An injectable melphalan composition (ALKERAN.RTM. for
Injection, GlaxoSmithKline) has been approved by the U.S. Food and
Drug Administration, for the palliative, treatment of patients with
multiple myeloma for whom oral therapy is not appropriate, and an
oral melphalan composition (ALKERAN.RTM. Tablets, GlaxoSmithKline)
has been approved for the palliative treatment of multiple myeloma
and for the palliation of non-respectable epithelial carcinoma of
the ovary.
[0004] ALKERAN.RTM. for Injection (GlaxoSmithKline) is administered
intravenously after first diluting a sterile, nonpyrogenic,
freeze-dried powder containing melphalan hydrochloride (equivalent
to 50 mg melphalan) and 20 mg povidone with a sterile diluent that
contains sodium citrate (0.2 g), propylene glycol (6 mL), ethanol
(96%, 0.52 mL), and water, for a total, volume of 10 mL. The usual
intravenous dose is 16 mg/m.sup.2, which is administered as a
single infusion over 15 to 20 minutes. Melphalan is intravenously
administered in 4 doses at 2-week, intervals, then, after adequate
recovery from toxicity, at 4-week intervals.
[0005] According to the ALKERAN.RTM. for Injection.
(GlaxoSmithKline) label, following administration of ALKERAN.RTM.
for Injection, drug plasma concentrations of melphalan decline
rapidly in a biexponential manner with distribution phase and
terminal elimination phase half-lives of approximately 10 and 75
minutes, respectively. The average total body clearance is 7 to 9
mL/min/kg (250 to 325 mL/min/m.sup.2). A study has reported that on
repeat dosing of 0.5 mg/kg every 6 weeks, the clearance of
melphalan decreased from 8.1 mL/min/kg after the first course, to
5.5 mL/min/kg after the third course, but did not decrease
appreciably after the third course. "Mean (.+-.SD) peak melphalan
plasma. concentrations in myeloma patients after administration of
10 or 20 mg/m.sup.2 doses of melphalan were 1.2.+-.0.4 and
2.8.+-.1.9 .mu.g/mL, respectively. After intravenous administration
of 50 mg of melphalan, the steady-state volume of distribution of
melphalan is 0.5 L/kg. The extent of melphalan binding to plasma
proteins ranges from 60% to 90%. Serum albumin, is the major
binding protein, while .alpha..sub.1-acid glycoprotein appears to
account for about 20% of the plasma protein binding. Approximately
30% of the drug is (covalently) irreversibly bound to plasma
proteins. Interactions with immunoglobulins have been found to be
negligible.
[0006] Melphalan is eliminated, from plasma primarily by chemical
hydrolysis to monohydroxymelphalan and dihydroxymelphalan. Aside
from these hydrolysis products, no other melphalan metabolites have
been observed in humans.
[0007] Controlled trials comparing intravenous to oral melphalan
have shown greater myelosuppression with the intravenously
administered melphalan. Furthermore; hypersensitivity reactions,
including anaphylaxis, have occurred in approximately 2% of
patients who have received intravenous melphalan. Melphalan also
undergoes rapid hydrolysis in aqueous solution. See S. A. Stout et
al., Int. J. Pharm. 24:193 (1985). Melphalan in the ALKERAN.RTM.
For Injection (GlaxoSmithKline) product also rapidly forms a
citrate derivative upon reconstitution and cannot be refrigerated,
due to precipitation of melphalan from solution.
[0008] Melphalan compositions comprising a cyclodextrin derivative
as a carrier and/or a diluent are known. See, e.g., Medlicott, et
al., J Pharm Sci. 87:1138 (1998), D. Q. Ma et al., Int. J. Pharm.
189:227 (1999), D. Q. Ma et al., J. Pharm. Sci. 89.275 (2000), and
U.S. Pat. Nos. 4,983,586, 5,024,998, and 6,583,125.
BRIEF SUMMARY OF THE INVENTION
[0009] What is needed is a melphalan formulation that can minimize
the toxicology and side-effect profile of intravenous melphalan.
What is also needed is an intravenous melphalan formulation having
increased bioavailability and/or an improved rate of therapeutic
onset. Also needed is a melphalan composition suitable for
intravenous administration that is stable under ambient and/or
refrigerated conditions, and can provide fully dissolved melphalan
without the need for organic solubilizers (e.g., ethanol and/or
propylene glycol, and the like). What is also needed is a
composition free from components that rapidly form a derivative
with melphalan. What is also needed is a melphalan composition
suitable for intravenous administration that has improved
stability, thereby enabling longer duration infusions, and
lengthening the melphalan exposure time that a patient receives
from a Single, convenient administration. As described herein,
compositions suitable for oral or parenteral administration, that
include melphalan and a cyclodextrin derivative have been
developed.
[0010] The present invention is directed to a pharmaceutical
composition comprising 25 mg to 125 mg of melphalan as a
hydrochloride salt, an optional buffer, and a cyclodextrin
derivative of formula I.
##STR00001##
[0011] wherein n is 4, 5 or 6; wherein. R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9
are independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein, at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and is a straight-chain or
branched C.sub.1-C.sub.8,-(alkylene)-SO.sub.3 group; wherein the
pharmaceutical composition has a pH of about 4 to about 6; wherein
dilution of the pharmaceutical composition with an aqueous solution
provides a melphalan solution ready for infusion in which the
melphalan degrades by 2% or less at about 25.degree. C. within 10
hours, or 4% or less at about 25.degree. C. within 10 hours after
the dilution; and wherein the cyclodextrin derivative is present in
a ratio of 50:1 to 100:1 (w/w) relative to the melphalan.
[0012] The present invention is also directed to a pharmaceutical
composition comprising 150 mg to 250 mg of melphalan as a
hydrochloride salt, an optional buffer, and a cyclodextrin
derivative of formula I:
##STR00002##
[0013] wherein n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C1-C8-(alkylene)-SO.sub.3 group, or an optionally substituted
straight-chain or branched C.sub.1-C.sub.6 group; wherein at least
one of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 is a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
pharmaceutical composition has a pH of about. 4 to about 6, wherein
dilution of the pharmaceutical composition with an aqueous:
solution provides, a melphalan solution ready for infusion in which
the melphalan degrades by 2% or less at about 25.degree. C. within
5 hours, or 4% or less at about 25.degree. C. within 10 hours after
the dilution, and wherein the cyclodextrin derivative is present in
a ratio of 25:1 to 35:1. (w/w) relative to the melphalan.
[0014] In some embodiments, the cyclodextrin derivative is a
compound of formula II:
##STR00003##
wherein. R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1.
[0015] In some embodiments, the cyclodextrin derivative is a
compound of formula II:
##STR00004##
wherein R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1;
and
[0016] and the pharmaceutical composition comprises about 50 mg of
melphalan as a hydrochloride salt and the cyclodextrin derivative
is present in a concentration of 50:1 to 100:1 (w/w) relative to
the melphalan; or
[0017] the pharmaceutical composition comprises about 50 mg of
melphalan as a hydrochloride salt the cyclodextrin derivative is
present in a ratio of about 55:1 (w/w) relative to the melphalan;
or
[0018] the pharmaceutical composition comprises about 200 mg of
melphalan as a hydrochloride salt and the cyclodextrin derivative
is present in a ratio of 25:1 to 35:1 (w/w): relative to the
melphalan; or
[0019] the pharmaceutical composition comprises about 200 mg of
melphalan as a hydrochloride salt and the cyclodextrin derivative
is present in a ratio of about 27:1, about 30:1, or about 32:1
(w/w) relative to the melphalan.
[0020] The present invention is directed to a method of treating a
subject suffering from a neoplastic disorder, the method comprising
diluting a composition with an aqueous diluent to provide a dilute
pharmaceutical composition comprising 25 mg to 125 mg of melphalan
and a cyclodextrin derivative of formula 1:
##STR00005##
[0021] wherein n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is a straight-chain
or branched. C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
dilute pharmaceutical composition has a pH of about 4 to about 6;
wherein the cyclodextrin derivative is present in a concentration
of at least 50:1 (w/w) relative to the melphalan; wherein the
melphalan in the dilute pharmaceutical composition degrades by 2%
or less at about 25.degree. C. within 5 hours, or 4% or less at
about 25.degree. C. within 10 hours after the diluting; and
administering the dilute pharmaceutical composition by injection to
the subject in need thereof.
[0022] In some embodiments, the neoplastic disorder is selected
from: myeloma, multiple myeloma, acute myelogenous leukemia,
melanoma, malignant melanoma, breast cancer, ovarian cancer,
testicular cancer, advanced prostate cancer, a neuroendocrine
cancer, metastatic melanoma (e.g., metastatic ocular melanoma,
metastatic cutaneous melanoma, and the like), a metastatic
neuroendocrine tumor, a metastatic adenocarcinoma tumor,
hepatocellular carcinoma, osteogenic sarcoma, polycythemia
veraplasma, plasma cell neoplasm, amyloidosis, sclerormyxedema, and
combinations thereof. In some embodiments, the neoplastic disorder
is multiple myeloma and the administering is systemic and provides
palliative treatment of the multiple myeloma.
[0023] The present invention is also directed. to a method for
conditioning a subject in need of a stem cell transplantation, the
method, comprising administering a melphalan dose of 50 mg/m.sup.2
to 300 mg/m.sup.2 per clay to the subject in need of the stem cell
transplantation, wherein the melphalan dose is administered in a
pharmaceutical composition comprising melphalan and a cyclodextrin
derivative of formula I:
##STR00006##
[0024] wherein n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is a straight-chain
or branched. C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
pharmaceutical composition has a pH of about 4 to about 6; and
wherein the cyclodextrin derivative is present in a ratio of at
least 25:1 (w/w) relative to the melphalan.
[0025] In some embodiments, at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R is a
hydroxy-substituted-C.sub.3 group.
[0026] In some embodiments, R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are independently a
straight-chain, or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3--
group having a degree of substitution of 4 to 8 per cyclodextrin
derivative, and the remaining substituents are --H.
[0027] In some embodiments, at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is
substituted with a straight-chain C.sub.4-(alkylene)-SO.sub.3--
group.
[0028] In some embodiments, a pharmaceutical composition or a
dilute pharmaceutical composition is substantially free of an
alcohol.
[0029] In some embodiments, the aqueous diluent is a saline
solution.
[0030] In some embodiments, a. dilute pharmaceutical composition is
stored about 0.5 hours to about 48 hours prior to the
administering. In some embodiments, the melphalan in a
pharmaceutical composition of the present invention degrades by 2%
or less at about 25.degree. C. within 5 hours, or 4% or less at
about 25.degree. C. within 10 hours after the diluting.
[0031] In some embodiments, a subject suffering from a neoplastic
disorder or in need of a stem cell transplantation is a pediatric
subject.
[0032] In some embodiments, the administering is performed
intravenously. In some embodiments, the administering is performed
via a limb perfusion.
[0033] In some embodiments, the administering is for a period of
two or more days.
[0034] In some embodiments, the administering provides a melphalan
C.sub.max in a subject that is at least 20% or greater than a
melphalan C.sub.max provided by a melphalan formulation containing,
an equivalent dose of melphalan and lacking the cyclodextrin
derivative. In some embodiments, the administering provides a
melphalan AUC.sub.0-1 in a subject that is at least 20% or greater
than a melphalan AUC.sub.0-1 provided by a melphalan formulation
containing an equivalent dose of melphalan and lacking the
cyclodextrin derivative.
[0035] In some embodiments, a method of the present invention
comprises diluting a concentrated melphalan composition with an
aqueous diluent to provide the pharmaceutical composition. In some
embodiments, a concentrated melphalan composition, comprises 50 mg
to 500 mg melphalan. In some embodiments, a concentrated melphalan
composition comprises about 200 mg of melphalan.
[0036] The present invention is also directed to a pharmaceutical
kit comprising a first container comprising 25 mg to 125 mg of
melphalan as a hydrochloride salt and an optional water-soluble
polymer, and a second container comprising an aqueous diluent, an
optional buffer, and a cyclodextrin derivative of formula I.
##STR00007##
[0037] wherein la is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, as Straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally.
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is a straight-chain
or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
cyclodextrin derivative is present in the second container in a
concentration of at least 50.1 (w/w) relative to the melphalan; and
wherein combining the first container and the second container
provides a dilute pharmaceutical composition having a pH of about 4
to about 6 that degrades by 2% or less at about 25.degree. C.
within 5 hours, or 4% or less at about 25.degree. C. within 10
hours after the diluting.
[0038] The present invention is also directed to a. pharmaceutical
kit comprising a first container comprising 150 mg to 250 mg of
melphalan as a hydrochloride salt and an optional water-soluble
polymer; and a second container comprising an aqueous diluent; an
optional buffer, and a cyclodextrin derivative of formula I:
##STR00008##
[0039] wherein n is 4, 5 or 6; Wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is a straight-chain
or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
cyclodextrin derivative is present in the second container in a
Concentration of 25:1 to 35:1 (w/w) relative to the melphalan; and
wherein combining the first container and the second container
provides a dilute pharmaceutical composition having a pH of about 4
to about 6 that degrades by 2% or less at about 25.degree. C.
within 5 hours, or 4% or less at about 25.degree. C. within 10
hours after the diluting.
[0040] In stone embodiments, a first container comprises povidone
in an amount of 10 mg to 30 mg, and a second container comprises a
pH-adjusting agent in a concentration sufficient to provide a pH of
about 4 to about 6 when, the. first container and the second
container are combined.
[0041] In some embodiments, the cyclodextrin derivative present in.
the second container is a compound of formula II:
##STR00009##
[0042] wherein. R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1;
wherein the first container comprises about 200 mg of melphalan as
a hydrochloride salt; and, wherein the cyclodextrin derivative is
present in the second container in an amount of about 27:1, about
30:1, or about 32:1 (w/w) relative to the melphalan.
[0043] Further embodiments, features, and advantages of the present
inventions, as well as the composition, structure and operation of
the various embodiments of the present invention, are described in
detail below with reference to the accompanying drawings.
BRIEF DESCRIPTION OF THE FIGURES
[0044] The accompanying drawings, which are incorporated, herein
and form a part of the specification, illustrate one or more
embodiments of the present invention and, together with the
description, further serve to explain the principles, of the
invention and to enable a person skilled in the pertinent art to
make and use the invention.
[0045] FIG. 1 provides a graphic representation of the solubility
of free base melphalan as a function of pH and the concentration of
a. cyclodextrin derivative.
[0046] FIG. 2 provides a graphic representation of the solubility
of free base melphalan and melphalan hydrochloride at pH 7.5 as a
function of the concentration of a cyclodextrin derivative.
[0047] FIGS. 3. and 4 provide flow charts that describe processes
for preparing a unit dosage form of the present invention.
[0048] FIGS. 5A-5B provide a graphic representation of
dose-normalized whole. blood (FIG. 5A) and plasma (FIG. 5B)
concentrations of melphalan following intravenous administration to
male Sprague-Dawley rats using a melphalan formulation containing a
Cyclodextrin derivative (SBE.sub.6:5-.beta.-CD) and a
cyclodextrin-free melphalan formulation (ALKERAN.RTM. for
Injection, GlaxoSmithKline).
[0049] FIG. 6 provides a graphic representation of the mean plasma
melphalan concentration in a human patient after intravenous
administration of a melphalan formulation containing a cyclodextrin
derivative (SBE.sub.6.5 .beta.-CD) and after intravenous
administration of a cyclodextrin-free melphalan formulation
(Melphalan HCl Injectable, Bioniche Pharma USA).
[0050] One or more embodiments of the present invention will now be
described with reference to the accompanying drawings. In the
drawings, like reference numbers can indicate identical or
functionally similar elements. Additionally; the left-most digit(s)
of a reference number can identify the drawing in which the
reference number first: appears.
DETAILED DESCRIPTION OF THE INVENTION
[0051] This specification discloses one or more embodiments that
incorporate the features of this invention. The disclosed
embodiment(s) merely exemplify the invention. The scope of the
invention is not limited to the disclosed embodiment(s). The
invention is. defined by the claims appended hereto.
[0052] Throughout the specification, use of the term "about" with
respect to any quantity is contemplated to include that quantity:
For example; "about 10 mL" is contemplated herein to include "10
mL," as well, as values understood in the art to be approximately
10 mL. with respect to the entity described.
[0053] The invention includes combinations and sub-combinations of
the various aspects and embodiments disclosed herein. Further, when
a particular feature, structure, or characteristic is described in
connection with an embodiment, it is understood that it is within
the knowledge of one skilled in the art to effect such feature,
structure, or characteristic in connection with other embodiments
whether or not explicitly described. These and other aspects of
this invention will be apparent upon reference to the following
detailed description, examples, claims and attached figures.
Melphalan
[0054] The compositions, formulations and unit dosage forms of the
present invention comprise melphalan, which has the following
chemical structure:
##STR00010##
[0055] As used herein, the term "melphalan" refers to the L-isomer
of the above compound, 4-[bis(chloroethyl)amino]phenylalanine, as
well as addition salt, polymorphs, solvates, hydrates, dehydrates,
co-crystals, anhydrous, and amorphous forms thereof. Melphalan
contains a chiral atom, and thus, as used herein, "melphalan" can
refer to the substantially pure form of the L-isomer. As used
herein, "substantially pure" refers to melphalan having a purity of
90% or higher, 95% or higher, 98% or higher, 99% or higher, 99.5%
or higher, or 99.9% or higher.
[0056] The D-isomer of the above compound, known as melphalan, is
less active against certain animal tumors, and the dose needed to
produce effects on chromosomes is larger than that required with
melphalan. The racemic (DL-) form is known as melphalan or
sarcolysin. In some embodiments, the compositions of the present
invention are substantially free of melphalan. In some embodiments,
the compositions of the present invention comprise melphalan as a
hydrochloride salt having a purity of 95% or greater, 98% or
greater, 99% or greater, 99.9% or greater, or 99.99% or
greater.
[0057] Melphalan is a bifunctional alkylating agent that is active
against selected human neoplastic diseases. The molecular formula
for melphalan is C.sub.13H.sub.18Cl.sub.2N.sub.2O.sub.2, and the
molecular weight of the free base form is 305.20 g/mol. Melphalan
is practically insoluble in water (pH 7) and has a pKa, of about
2.5.
[0058] In some embodiments, the pharmaceutical compositions and
dosage forms of the present invention comprise melphalan as a
hydrochloride salt. As used herein, "melphalan as a hydrochloride
salt" refers to the hydrochloric acid addition salt of the above
compound. However, amounts and concentrations of melphalan are
provided in reference to an, equivalent mass of free base
melphalan. Thus, 5 mg of "melphalan as a hydrochloride salt" refers
to 5 mg of the active agent melphalan, exclusive, of the
hydrochloride addition salt, which: if considered would provide a
total mass of about 5.6 mg.
[0059] Cyclodextrin Derivatives
[0060] The compositions, formulations and/or unit dosage forms of
the present invention comprise a cyclodextrin derivative. As used
herein, "cyclodextrin derivative" refers to a cyclic
oligosaccharide comprising five or more .alpha.-D-glucopyranoside
units linked in, circular 1.fwdarw.4 configuration, and comprising
a substituent group attached to one or more of the glucopyranoside
units at the 2, 3 and/or 6 position(s) through an ether bond
(O--R--, where R refers to the substituent group).
[0061] In some embodiments, the cyclodextrin derivative s a
compound of formula I:
##STR00011##
[0062] wherein: n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently selected, from: --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, and an optionally
substituted straight-chain, or branched. C.sub.1-C.sub.6 group.
[0063] In some embodiments, cyclodextrin for use with the present
invention is selected based, upon an average degree of substitution
("ADS"), which, as used herein refers to the average number of
substituent groups per cyclodextrin molecule. The average degree of
substitution for cyclodextrin derivatives is described in detail in
WO 2009/018069, which is incorporated, herein by reference in its
entirety. As used herein, a cyclodextrin derivative composition for
use with the present invention is referred to by the following
notation: the substituent(s) are abbreviated (e.g., sulfobutyl
ether groups are abbreviated as "SBE") with a subscript denoting
the ADS of the substituent, and cyclodextrin structure is defined.
For example, a sulfobutyl ether-derivatized .beta.-cyclodextrin
composition having an ADS of 6.5 is referred to as
"SBE.sub.6.5-.beta.-CD." As a second example, a .beta.-cyclodextrin
composition comprising cyclodextrin molecules derivatized with both
sulfobutyl ether and hydroxypropyl groups is referred to as
"SBE.sub.4.2-HP.sub.2.5-.beta.-CD," wherein the ADS of the
sulfobutyl ether groups is 4.2 and the ADS of the hydroxypropyl
groups is 2.5.
[0064] Cyclodextrin derivatives suitable for use with the present
invention include cyclodextrin compositions bearing substituent
groups (R.sub.1-R.sub.9 and R in formulas I and II, respectively)
that are independently selected from: --H, a straight-chain or
branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, and an
optionally substituted straight-chain or branched C.sub.1-C.sub.6
group.
[0065] In some embodiments, at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 of
formula I is substituted with a straight-chain
C.sub.4-(alkylene)-SO.sub.3 group. Exemplary
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 groups suitable for use with
the present invention include, but are not limited to, sulfoethyl,
sulfopropyl, 1-methyl-sulfopropyl, sulfobutyl, 1-methyl-sulfobutyl,
2-methyl-sulfobutyl, 1-methyl-sulfobut-3-yl, 2-ethyl-sulfobutyl,
3-ethyl-sulfobutyl, sulfopentyl, 1-sulfopent-3-yl, sulfohexyl,
sulfoheptyl, sulfooctyk and the like, and combinations thereof.
[0066] In some embodiments, R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 of formula I are
independently a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group having an ADS of 4 to 8,
4 to 7.5, 4 to 7, 4 to 6.5, 4.5 to 8, 4.5 to 7.5, 4.5 to 7, 5 to 8,
5 to 7.5, 5 to 7, 5.5 to 8, 5.5 to 7.5, 5.5 to 7, 5.5 to 6.5, 6 to
8, 6 7.5, 6 to 7.1, 6.5 to 7, about 6.5, or about 7 per
cyclodextrin derivative, and the remaining substituents are
--H.
[0067] In some embodiments, a substituent is an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group. As
used herein, "optionally substituted" refers to one or more
optional substituents selected from: halogen (i.e., --F, --Cl,
--Br, --I), --NO.sub.2, --C.dbd.N, --OR.sub.22, --SR.sub.22,
--SO.sub.2R.sub.22, --C(.dbd.O)OR.sub.22, --C(.dbd.O)R.sub.22,
--C(.dbd.O)N(R.sub.22).sub.2, --SO.sub.2N(R.sub.22).sub.2,
--SO.sub.2N(H)C(.dbd.O)R.sub.22, --SO.sub.2N(H)C(.dbd.O)OR.sub.22
(wherein R.sub.22 is not H), --N(R.sub.22).sub.2,
--N(R.sub.22)SO.sub.2R.sub.22, --N(R.sub.22)C(O).sub.mR.sub.22
(wherein m=1 or 2), --N(R.sub.22)C(O)N(R.sub.22).sub.2,
--N(R.sub.22)SO.sub.2N(R.sub.22).sub.2, --O--C(.dbd.O)R.sub.22,
--O--C(.dbd.O)OR.sub.22, --O--C(.dbd.O)N(R.sub.22).sub.2,
--C(.dbd.O)N(H)SO.sub.2N(R.sub.22).sub.2,
--C(.dbd.O)N(H)SO.sub.2R.sub.22, oxo (or keto, i.e., .dbd.O);
thioxo (i.e., .dbd.S), imino (i.e., .dbd.N(R.sub.22),
--NR.sub.22--C(.dbd.NR.sub.22)R.sub.22,
--NR.sub.22--C(.dbd.NR.sub.22)N(R.sub.22).sub.2,
--C(.dbd.NR.sub.22)N(R.sub.22).sub.2,
--O--C(.dbd.NR.sub.22)N(R.sub.22).sub.2,
--O--C(.dbd.NR.sub.22)R.sub.22, --C(.dbd.NR.sub.22)R.sub.22,
--C(.dbd.NR.sub.22)OR.sub.22, and ionic forms thereof (e.g.,
--N.sup.+(R.sub.22).sub.2X.sup.-, and the like, wherein X-- is a
pharmaceutically acceptable anion), wherein R.sub.22 is
independently selected at each occurrence from H, and
C.sub.1-C.sub.4 alkyl.
[0068] Exemplary optionally substituted straight-chain or branched
C.sub.1-C.sub.6 groups include, but are not limited to,
2-hydroxypropyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 3-oxobutyl,
and 2-ethoxy-ethyl.
[0069] In some embodiments, at least one of R.sub.1, R.sub.2,
R.sub.3, R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 of
formula I is a hydroxy-substituted-C.sub.3 group. In some
embodiments, the cyclodextrin derivative comprises
.beta.-cyclodextrin that includes, a hydroxy-substituted-C.sub.3
group having an ADS of 1 to 8, 2 to 8, 3 to 7, 4 to 7.5, 4.3 to
7.5, about 1, about 2, about 2.5, about 3, about 3.5, about 4,
about 4.3, about 5, about 5.5, about 6, about 6.5, about 7, or
about 7.5.
[0070] Exemplary cyclodextrin compositions, and methods of making
the same, that are suitable for use with the present invention also
include those described in U.S. Pat. Nos. 5,134,127, 5,241,059,
5,376,645, 5,874,418, 6,046,177, 6,133,248, 6,153,746, 6,204,256,
7,034,013, 7,629,331, and 7,635,773, U.S. Pub. No. 2009/0012042,
and PCT Pub. No. WO 2005/117911, the contents of each of which is
incorporated herein by reference, in the entirety.
[0071] In some embodiments, the cyclodextrin derivative is a
compound of formula II.
##STR00012##
[0072] wherein R(H).sub.21-x or
(--CH.sub.2).sub.4--SO.sub.3Na.sup.+).sub.x. In some embodiments,
x=6.0-7.1. In some embodiments, the cyclodextrin derivative of
formula II has an average molecular weight of about 2163 g/mol.
[0073] In some embodiments, the cyclodextrin derivative is a
sulfobutyl ether-.beta.-cyclodextrin having an ADS of about 7
(e.g., CAPTISOL.RTM., CyDex Pharmaceuticals, Inc., Lenexa, Kans.).
Captisol, cyclodextrin is a polyanionic .beta.-cyclodextrin
derivative with a sodium sulfonate salt separated from the
lipophilic cyclodextrin cavity by a butyl ether spacer group, or
sulfobutylether (SBE). CAPTISOL.RTM. cyclodextrin has been shown to
be safe when administered parenterally, orally, or via inhalation
and does not exhibit the nephrotoxicity associated with.
.beta.-cyclodextrin. Relative, to .beta.-cyclodextrin,
CAPTISOL.RTM. sulfoalkyl ether cyclodextrin provides comparable or
higher complexation characteristics and superior water solubility
in excess of 90 g per 100 mL, a 50-fold improvement. Melphalan has
a low binding affinity with CAPTISOL.RTM. (K.sub.a=3.times.10.sup.2
M.sup.1).
[0074] In some embodiments, the cyclodextrin derivative includes a
substituent that, bears an ionic group that can optionally form a
salt with a pharmaceutically acceptable anion or cation.
Pharmaceutically acceptable cations suitable for forming salts
with. negatively charged cyclodextrin derivatives of the present
invention include, but are not limited to, H.sup.+Li.sup.+,
Na.sup.+, K.sup.+, Mg.sup.2+, Ca.sup.2+, ammonium and amine cations
such as cations of (C.sub.1-C.sub.6) alkylamines,
(C.sub.4-C.sub.8)-cycloalkylamines (e.g., piperidine, pyrazine, and
the like), (C.sub.1-C.sub.6) alkanolamines, and
(C.sub.4-C.sub.8)-cycloalkanolamines, and the like; and
combinations thereof. In some embodiments, a pharmaceutically
acceptable cation is Na.sup.+. Pharmaceutically acceptable anions
suitable for forming salts with positively charged cyclodextrin
derivatives of the present invention include, but are not limited
to, halides (e.g., Cl.sup.- and the like), anions of
(C.sub.1-C.sub.6)-alkyl acids (e.g., acetate, oxalate, fumarate,
succinate, and the like, and combinations thereof.
[0075] Pharmaceutical Compositions and Unit Dosage Forms
[0076] The present invention is directed to pharmaceutical
compositions and unit dosage forms comprising, melphalan and a
cyclodextrin derivative. The pharmaceutical compositions of the
present invention are suitable for parenteral administration to a
subject. Parenteral administration of the pharmaceutical
compositions can include, but is not limited to, an injection.
Because parenteral administration bypasses a subjects natural
defenses against contaminants, the pharmaceutical compositions are
sterile, or capable of being sterilized prior to
administration.
[0077] Exemplary pharmaceutical compositions include, but are not
limited to, solutions, suspensions or emulsions ready for
administration, solutions, suspensions or emulsions ready to be
dissolved in and/or diluted with a pharmaceutically acceptable
vehicle, and dry products ready to be dissolved in and/or diluted
with a pharmaceutically: acceptable vehicle:
[0078] Generally, the pharmaceutical compositions of the present
invention comprise melphalan in a concentration suitable for
treating a condition that is amenable: to treatment with melphalan.
Thus, the pharmaceutical compositions of the present invention can
be used to prepare a unit dosage form comprising: a therapeutically
effective amount of melphalan for administering to a subject in
need thereof. In some embodiments, the present invention is
directed to a unit dosage form that comprises melphalan in a
concentration that is suitable for administration without dilution.
Alternatively, a unit dosage form of the present invention, can be
diluted prior to administration to a subject in need thereof.
[0079] The present invention is also directed to: a pharmaceutical
composition comprising 25 mg to 125 mg, 25 mg to 100 mg, 25. mg to
75 mg, 25 mg to 50 mg, 50 mg to 125 mg, 50 mg to 100 mg, 75 to 125
mg, 100 to 125 mg, about 25 mg, about 50 mg, about 75 mg, about 100
mg, or about 125 mg of melphalan as a hydrochloride salt, an
optional buffer, and a cyclodextrin derivative of formula I:
##STR00013##
[0080] wherein n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein, at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is a straight-chain
or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
pharmaceutical composition has a pH of about 4 to. about 6, about 4
to about 5, about 4.5 to about 6, about 5 to about 6, about 5.5 to
about 6, about 4, about 4.5, about 5, about 5:5, or about 6;
wherein dilution of the pharmaceutical composition with an aqueous
solution provides a solution in which the melphalan degrades by 2%
or less at about 25.degree. C. within 5 hours, or by 4% or less at
about 25.degree. C. within 10 hours after the dilution; and wherein
the cyclodextrin derivative is present in a ratio: of 50:1 to
100:1, 55:1 to 60:1, about 50:1, about 55:1, or about 60:1 (w/w)
relative to the melphalan.
[0081] The present: invention is also directed to. a pharmaceutical
composition comprising 150 mg to 300 mg, 150 mg to 250 mg, 150 mg
to 225 mg, 175 mg to 250 mg, 200 mg to 250 mg, about 150 mg, about
175 mg, about 200 mg, about 225 mg, or about 250 mg of melphalan as
a hydrochloride salt, an optional buffer, and a cyclodextrin
derivative of formula I:
##STR00014##
[0082] wherein n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7; R.sub.8 and R.sub.9 is a straight-chain
or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein the
pharmaceutical composition has a pH of about 4 to about 6, wherein
dilution of the pharmaceutical composition with a aqueous solution
provides a melphalan solution in Which the melphalan degrades by 2%
or less at about 25.degree. C. within 5 hours, or 4% or less at
about 25.degree. C. within 10 hours after the dilution; and wherein
the cyclodextrin. derivative is present in. a ratio of 25:1 to
35:1, about 0.27:1, about 30:1, or about 32:1 (w/w) relative to the
melphalan.
[0083] In some embodiments, the cyclodextrin derivative is a
compound of formula II:
##STR00015##
[0084] wherein R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4--SO.sub.3--Na).sub.x, and x=6.0-7.1,
[0085] In some embodiments, the cyclodextrin derivative is a
compound of formula II:
##STR00016##
[0086] wherein R.dbd.(H).sub.21-x or
(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1; and
[0087] the pharmaceutical composition comprises about 50 mg of
melphalan as a hydrochloride:salt and the cyclodextrin derivative
is present in a concentration of 50:1 to 100:1, 55:1 to 60:1, about
50:1, about 55:1, or about 60:1 (w/w relative to the melphalan;
or
[0088] the pharmaceutical. composition: Comprises about 200 mg of
melphalan as a hydrochloride salt and the cyclodextrin derivative
is present in a ratio of 25:1 to 35:1, about 27:1, about 30:1, or
about 32:1 (w/w) relative to the melphalan.
[0089] Sterile solutions, suspensions, emulsions and the like can
be prepared by incorporating melphalan into an appropriate solvent
or carrier with the other, optional ingredients enumerated herein,
followed by sterilization. Sterile powders can be prepared by spray
drying, aseptic spray drying, vacuum drying, or freeze drying a
sterile, solution, suspension, or emulsion to' provide a dried
solid (e.g., a powder) comprising melphalan along with any
additional excipients.
[0090] In some embodiments, the present invention is directed to a'
solid pharmaceutical composition consisting of about 50 mg of
melphalan as a hydrochloride salt, an amount sufficient of an acid,
a. base, or a combination thereof to provide, a pH of about 4 to
about 6 upon dilution with a saline solution to a volume of about
10 mL, and a cyclodextrin derivative of formula II:
##STR00017##
[0091] wherein R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1,
wherein dilution of the solid pharmaceutical composition with an
aqueous solution provides a melphalan solution in which the
melphalan degrades by 2% or less at about 25.degree. C. within 5
hours, or by 4% or less at about 25.degree. C. within 10 hours
after the diluting, and wherein the cyclodextrin derivative is
present in a ratio of about 55:1 (w/w) relative to the
melphalan.
[0092] In some embodiments, the present invention is directed to a
solid pharmaceutical composition consisting of about 200 mg of
melphalan as a hydrochloride: salt, an amount sufficient of an
acid, a base, or a combination thereof to: provide a pH of about 4
to about 6 upon dilution with a saline: solution to a volume of
abut. 20 mL, and a cyclodextrin derivative of formula II:
##STR00018##
[0093] wherein R.dbd.(H).sub.21-x or
(--(CH.sub.2).sub.4--SO.sub.3--Na.sup.+).sub.x, and x=6.0-7.1,
wherein dilution of the solid pharmaceutical composition with an
aqueous solution provides a melphalan solution in which the
melphalan degrades by 2% or less at about 25.degree. C. within 5
hours, or by 4% or less at about 25.degree. C. within 10 hours
after the diluting, and wherein the cyclodextrin derivative is
present in a ratio: of about 27:1, about 30:1, or about 32:1 (w/w)
relative to the melphalan.
[0094] In some embodiments, a pharmaceutical composition: or unit
dosage form of the present invention comprises a solid (e.g., a
powder) or a liquid solution that is diluted with a liquid carrier
or diluent prior to administration to a subject. Thus, the
pharmaceutical compositions: and unit dosage forms of the present
invention include sterile aqueous solutions, suspensions and
dispersions, as well as sterile solids (e.g., powders) comprising
melphalan that can be extemporaneously diluted or solubilized to
provide a sterile solution, suspension or dispersion.
[0095] In some embodiments, the compositions, formulations and/or
unit dosage forms of the present invention comprise a
pharmaceutically acceptable excipient. As used herein,
"pharmaceutically acceptable" refers to those excipients,
compounds, materials, and/or compositions which are, within the:
scope of sound medical judgment, suitable for contact with the
tissues of human beings and animals without excessive toxicity,
irritation, allergic response, or other possible complications
commensurate with a reasonable benefit/risk ratio.
[0096] In some embodiments, the: pharmaceutical compositions and
unit dosage forms of the present invention are substantially
homogeneous. As used herein, "homogeneous" refers to mixtures,
solutions, suspensions, compositions, dosage forms, and/or
formulations of the present invention that have a uniform
distribution of ingredients throughout. Homogeneity is synonymous
with uniformity and can refer to intra-sample uniformity,
batch-to-batch uniformity, run-to-run uniformity, and/or dosage
form-to-dosage form uniformity. For example, intra-sample
uniformity can be determined by analyzing a first: portion of a
sample,: Mixture, or composition and comparing this with a second
portion of the same sample, mixture, or composition. Typical
deviations of a composition (e.g., variation in the percentage by
weight of excipients: and the like) of a substantially homogeneous
composition are about 5% or less, about 3% or less, about 2% or
less; about 1% or less, or within experimental error.
[0097] In some embodiments, a pharmaceutical composition or unit
dosage form of the present invention comprises a pharmaceutically
acceptable excipient. As: used herein, the term "excipient" refers
to any inert. substance that can, be combined with melphalan and
the sulfoalkyl ether cyclodextrin for preparing the pharmaceutical
compositions.
[0098] Pharmaceutically acceptable excipients suitable for use with
the present. invention include, but are not limited to, a carrier;
a water-soluble polymer, a preservative, an antioxidant, a
pH-adjusting agent (e.g., an acidifying, agent, an alkalinizing
agent, and/or a buffer), a bulking. agent, a complexation enhancing
agent, a cryoprotectant, a density modifier, an electrolyte, a
flavor, a fragrance, a lyophilizing aid (e.g., a bulking agent
and/or stabilizing agent), a plasticizer, a solubility-enhancing
agent, a stabilizing, agent, a sweetener, a surface tension
modifier, a volatility modifier, a viscosity modifier, and
combinations thereof. In addition, one of skill in the art will
recognize that pharmaceutically acceptable excipients can be used
in the present invention, including those listed in The Handbook of
Pharmaceutical Excipients, 5th Ed., The Pharmaceutical Press and
American Pharmacists Association, London, U K and Washington, D.C.
(2006), which is incorporated herein by reference in its
entirety.
[0099] In some embodiments, a pharmaceutical composition or unit
dosage form of the present invention comprises a pharmaceutically
acceptable carrier. As used herein, a "carrier" refers to a vehicle
suitable for transferring and/or diluting a pharmaceutical
composition or unit dosage form of the present invention.
Pharmaceutically acceptable carriers suitable for use with the
present Inventions include, but are not limited to, liquids,
solids, colloids, gels, and combinations thereof. Liquid carriers
suitable for use with the present invention include solvents,
liquid dispersion mediums, and the like, such as, but not limited
to, water, ethanol, a polyol (e.g., glycerol, propylene glycol,
liquid polyethylene glycols, and the like), a vegetable, oil, a
nontoxic glyceryl ester, and combinations thereof. In some
embodiments, a liquid carrier is selected from a dextrose solution,
a saline solution, plasma, and lactated Ringer's solution.
[0100] In some embodiments, a pharmaceutical composition or unit
dosage form of the present invention comprises a water-soluble
polymer such as, but not limited to, homopolymers of
N-polyvinylpyrrolidone (e.g., "povidone"), low molecular weight
hydroxypropyl cellulose, low molecular weight methyl cellulose, low
molecular weight hydroxypropyl methyl cellulose, and the like, and
combinations thereof.
[0101] In some embodiments, after dilution a cyclodextrin
derivative is present in the diluted pharmaceutical composition in
a concentration of about 75 mM, about 100 mM, or about 125 mM.
[0102] In some embodiments, after dilution melphalan is present in
the diluted pharmaceutical composition in a: concentration of 0.1
mg/mL to 50 mg/mL, 0.15 mg/mL to 40 mg/mL, 0.2 mg/mL to 30 mg/mL,
0.3 mg/mL to 25 mg/mL, 0.4 mg/mL to 20 mg/mL, 0.45 mg/mL to 15
mg/mL, 0.5 mg/mL to 10 mg/mL, about 0.45 mg/mL, about 1 mg/mL,
about 1.5 mg/mL, about 2 mg/mL, about 2.5 mg/mL, or about 5
mg/mL.
[0103] In some embodiments, a pharmaceutical composition and/or a
diluent for use with a composition of the present invention, is
free of a solubilizing agent such as, but not limited to, water, an
alcohol (e.g., ethanol and the like), a polyol (e.g., glycerol,
propylene glycol, liquid polyethylene glycols, and the like), a
vegetable oil, a nontoxic glyceryl ester, and combinations thereof.
Thus, in some embodiments the diluent consists essentially of water
and optional. tonicity-adjusting agents (e.g., 0.9% saline solution
for injection, and the like).
[0104] In some embodiments, the pH of a pharmaceutical composition
or unit. Dosage form is controlled. In some embodiments, a
pharmaceutical composition or unit dosage form of the present.
invention comprises a pharmaceutically acceptable buffer and/or pH.
adjusting agent (e.g., an. acidifying agent. and/or alkalinizing
agent). In some embodiments, a pharmaceutical composition or unit
dosage. form of the present invention has a pH of about 4 to about
6, about 4 to about 5, about 5 to about 6, about 4, about 5, about
5.5, or about 6 after dilution with an aqueous diluent.
[0105] In some embodiments, a pharmaceutical composition or unit
dosage form that is to be diluted prior to administration to a
subject has a pH of about 2 to about 6, about 3 to about 6, about 4
to about 6, or about 5 to about 6. In some embodiments, after
dilution with an liquid carrier) a unit: dosage form of the present
invention has a pH of about 4 to about 6, about 4 to about 5, about
5 to about 6, about 4, about 4.5, about 5, about 5.5, or about 6 at
the time of administration to a subject in need thereof.
[0106] In some embodiments, a pharmaceutical composition or unit
dosage form of the present invention comprises a buffer. In some
embodiments; a pharmaceutical composition or unit dosage form of
the present invention comprises a buffer suitable to provide a
dilute composition having a pH of about 4 to about 6, about 4 to
about 5, about 5 to about 6, about 4, about 4.5, about 5 about 5.5,
or about 6. In some embodiments, a buffer is present in a
concentration of about 0.01 M to about 10 M, about 0.01 M to about
5 M, or about 0.01 M to about 1 M.
[0107] In some embodiments, a pharmaceutical composition or unit
dosage form of the present: invention comprises a pH-adjusting
agent such as, but not limited to, an acidifying agent (e.g.,
citric acid, HCl, and the like), an alkalinizing agent (e.g., NaOH
and the like), a salt form of an acid (e.g., sodium citrate and the
like), and combinations thereof. In some embodiments, a
pharmaceutical composition or unit dosage form of the: present
invention comprises a pH-adjusting agent in an amount sufficient to
provide dilute composition having a pH of about 4 to about 6, about
4 to about 5, about 5 to about 6, about 4, about 4.5, about 5
about. 5.5, or about 6. In some embodiments, In some embodiments, a
pharmaceutical composition or unit dosage form, of the present
invention comprises sodium citrate in an amount of 50 mg to 500 mg,
75 mg to 400 mg, 100 mg to 300 mg, 150 mg to 250 mg, or about 200
mg.
[0108] In some embodiments, a pharmaceutical composition or unit
dosage form of the present, invention comprises a second
therapeutic agent. Suitable second therapeutic agents include, but
are not limited to, a platinum compound, an antimetabolite, a
nitrosourea, a corticosteroid, a calcineurin inhibitor, a
monoclonal antibody, a polyclonal antibody, a cytotoxic antibiotic,
an interferon, an opioid, an antihistamine, a volume expander, a
pressor agent, and combinations thereof. Additional second
therapeutic agents include, but are not limited to, doxorubicin,
bortezomib, rituximab, thalidomide, lenalidomide, gemcitabine,
thiotepa, fludarabine, carmustine, etoposide, cytarabine,
granulocyte colony-stimulating factor, ADH-1, topotecan,
palifermin, prednisone, arsenic trioxide, ascorbic acid, busulfan,
buthionine sulfoximine, and combinations thereof.
[0109] As used herein, a "unit dosage form" refers to a composition
containing a specific amount of melphalan, the whole of which is
intended, to be administered to a subject in a single dose: A unit
dosage form: can be distinguished from a supply of a multi-dose
amount of a pharmaceutical composition, e.g., a bottle of medicine,
from which a unit dose is measured out.
[0110] In some embodiments, a. unit dosage form of the present
invention comprises a therapeutically effective amount of
melphalan. As used. herein, a "therapeutically effective amount"
refers to an amount of melphalan that, elicits a. biological or
medicinal response in a tissue, system, animal or human that is
being sought by a researcher, veterinarian, medical doctor or other
clinician, which includes alleviation of the symptoms of a disease
or disorder being treated.
[0111] A unit dosage form typically comprises the pharmaceutical
composition of the present invention and optionally, one or more
pharmaceutically acceptable excipients, wherein the amount of
melphalan present in the unit dosage form is sufficient for a
single administration to. a subject in. need thereof. Unit dosage
forms of the present invention include, but are not limited to,
liquid solution, liquid suspensions, liquid dispersions, emulsions,
gels, powders, tablets, capsules, caplets, and the like. Treatment
of a disease or condition amenable to treatment with melphalan can
comprises periodic administration of a unit dosage form of the
present invention, for example, once every two weeks, once every
four weeks, or some, other interval.
[0112] In some embodiments, a unit dosage form of the present
invention comprises 25 mg to 125 mg, or 150 mg to 250 mg of
melphalan as a hydrochloride salt. In some embodiments, a unit
dosage form of the present invention comprises 50 mg or 200 mg
melphalan as a hydrochloride Salt.
[0113] In some embodiments, a unit dosage form of the present
invention is a solid. In some embodiments, a solid unit, dosage
form of the present invention is a lyophilized solid, or an aseptic
spray-dried solid. In some embodiments, a dosage form of the
present invention is suitable for dilution and/or reconstitution
with a predetermined amount of a liquid carrier: For example, a
unit dosage form (e.g., as liquid or a solid) of the present
invention can be diluted with 5 mL to 500 mL, 10 mL to 100 mL, or
10 mL, to 50 mL of a liquid carrier.
[0114] The pharmaceutical compositions: and. unit dosage forms of
the present invention are stable. As used herein, stability can
refer to. either the shelf-life of an undiluted solid or liquid
dosage form or the resistance to degradation of a diluted liquid
dosage form. In particular; currently available melphalan
compositions suitable for intravenous administration must be used
as soon as possible after dilution due to the rapid degradation of
melphalan in aqueous solution. However, the dosage forms of the
present invention are stable for a considerable time period after
dilution, for example; at least 90 minutes up to at least 48 hours
or more. Thus, in those embodiments in which a solid or liquid unit
dosage form is diluted, the diluting can be performed immediately
prior to administering, or sometime before the administering
without any significant loss of therapeutic efficacy. This enables
a liquid pharmaceutical composition or liquid unit dosage form of
the present invention to be diluted 90 minutes to 48 hours in
advance of use (i.e., in advance of parenteral administration to a
subject in need thereof).
[0115] In some embodiments, the melphalan in a pharmaceutical
composition of the present invention degrades by 2% or less at
about 25.degree. C. within 5 hours, or by 4% or less at about
25.degree. C. within 10 hours after dilution with an aqueous
diluent to provide a diluted composition comprising a cyclodextrin
derivative in a concentration of about 75 mM or about 125 mM.
[0116] The primary degradation, product of melphalan in aqueous
solution is melphalan monohydroxide (also known as
monohydroxyrnelphalan), which proceeds via a hydrolysis reaction.
See, e.g., S. A. Stout et al., Int Pharm. 24:1.93 (1985). In some
embodiments, dilution of a pharmaceutical composition of the
present invention provides a melphalan monohydroxide concentration
(based on a 100% initial concentration. of melphalan) of 2% or less
within 5 hours of the diluting, when the diluted composition is
maintained at room temperature (about 25.degree. C.). In some
embodiments, dilution of a pharmaceutical composition of the
present invention provides a melphalan monohydroxide concentration
(based on 100% initial concentration of melphalan) of 4% or less
within 10 hours of the diluting, when the diluted composition is
maintained at room temperature (about 25.degree. C.). In some
embodiments, dilution of a pharmaceutical composition of the.
present invention provides a melphalan monohydroxide concentration
(based on a 100% initial concentration of melphalan) of 2% or less
within 24 hours of the diluting, or 4% or less within 48 hours of
the diluting when the diluted composition is maintained at a,
temperature of about 10.degree. C. or less.
[0117] Furthermore, the Pharmaceutical compositions of the present
invention can be stored prior to dilution for an extended period of
time without any significant loss of melphalan. For example, a
solid pharmaceutical composition comprising melphalan and a
cyclodextrin derivative contains 2% or less, by weight, of a
melphalan degradant after storage at 25.degree. C. for a period of
at least 2 years, or 5% or less, by weight, of a melphalan
degradant after storage at 25.degree. C. for a period of at least 3
years.
[0118] In some embodiments, a dry powder pharmaceutical composition
of the present invention forms 2% or less of melphalan
monohydroxide (based on a 100% initial concentration of melphalan)
after storage for 2 years at room temperature.
[0119] Pharmaceutical Kits
[0120] The present invention is also directed to a pharmaceutical
kit comprising a first container comprising 25 mg to 125 mg of
melphalan as a hydrochloride salt and an optional water-soluble
polymer, and a second container comprising an aqueous diluent, an
optional buffer, and a cyclodextrin, derivative of formula I.
##STR00019##
[0121] wherein n s 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H; a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group, or an optionally
substituted straight-chain or branched C.sub.1-C.sub.6 group;
wherein at least one of R.sub.1, R.sub.2, R.sub.3, R.sub.4,
R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 is a straight-chain
or branched C.sub.1-C.sub.8-(alkylene)-SO.sub.3 group; wherein, the
cyclodextrin derivative is present in the, second container in a
concentration of at least 50:1 (w/w) relative to the melphalan; and
wherein combining the. first container and the second container
provides a dilute pharmaceutical composition having a pH of about 4
to about 6 that degrades by 2% or less at about 25.degree. C.
within 5 hours after the diluting.
[0122] The present invention is also directed to a pharmaceutical
kit comprising a first container comprising 150 mg to 250 mg of
melphalan as a hydrochloride salt and an optional water-soluble
polymer; and a second container comprising an aqueous diluent, an
optional buffer, and a cyclodextrin derivative of formula I:
##STR00020##
[0123] wherein n is 4, 5 or 6; wherein R.sub.1, R.sub.2, R.sub.3,
R.sub.4, R.sub.5, R.sub.6, R.sub.7, R.sub.8 and R.sub.9 are
independently --H, a straight-chain or branched
C.sub.1-C.sub.8-(alkylene)-SO3- group, or an optionally substituted
straight-chain or branched C.sub.1-C.sub.6 group; wherein at least
one of R.sub.1, R.sub.2, R.sub.3, R.sub.4, R.sub.5, R.sub.6,
R.sub.7, R.sub.8 and R.sub.9 is a straight chain or branched
C1-C8-(alkylene)-SO.sub.3 group; wherein the cyclodextrin
derivative is present in the second container in a concentration of
25:1 to. 35:1 (w/w) relative to the melphalan; and wherein
combining the first container and the second container provides a
dilute: pharmaceutical composition having a pH of about 4 to about
6 that degrades by 2% or less at about 25.degree. C. within 5 hours
after the diluting.
[0124] Alternatively, a first container comprises melphalan (in an
amount described above) and a cyclodextrin derivative (as described
above), an optional water-soluble polymer (e.g., povidone and the
like), and an optional pH-adjusting agent; and a second container
comprises a diluent (e.g., water, saline, and the like), an
optional tonicity adjusting agent, and an optional pH-adjusting
agent.
[0125] Materials suitable for use as the containers with the kits
of the present invention include, but are not limited to, a glass
(e.g., borosilicate glass, amber glass, and the like), a plastic
(e.g., polypropylene, high-density polyethylene, poly(ethylene
terephthalate, polystyrene, polycarbonate, and the like, and
combinations thereof), a metal (e.g., a foil), and the like, and
combinations thereof (e.g., a plastic-coated glass and/or
metal).
[0126] Containers suitable for use with the pharmaceutical kits of
the present invention include, but are not limited to, vials,
bottles, sachets, and the like. The containers can be opened and/or
the contents can be removed therefrom, by, for example, tearing,
cutting, removing a screw-top, removing a stopper, piercing,
squeezing, and the like, and combinations thereof.
[0127] In some embodiments, a first container comprises povidone in
an amount of 10 mg to 30 mg, 15 mg to 25 mg, or about 20 mg. In
some embodiments, a second container comprises a pH-adjusting agent
(e.g., an acidifying agent, an alkalinizing agent, and/or a buffer)
in a concentration sufficient to provide a pH of about 4 to about 6
when the first container and the second container are combined. In
some embodiments, a second container comprises sodium citrate in an
amount of 5.0 mg to 500 mg, 75 mg to 400 mg, 100 mg to 300 mg, 150
mg to 250 mg, or about 200 mg.
[0128] In some embodiments, the cyclodextrin derivative present in
the second container is a compound of formula II:
##STR00021##
[0129] wherein R.dbd.(H).sub.21-x. or
(--(CH.sub.2).sub.4--SO.sub.3Na.sup.+).sub.x, and x=6.0-7.1;
wherein the first container comprises about 200 mg of melphalan as
a hydrochloride salt; and wherein the cyclodextrin derivative is
present in the second container in an amount of about 27:1, about
30:1, or about 32:1 (w/w) relative to the melphalan.
[0130] Methods of Administering and Treating
[0131] In some embodiments, the present invention is directed to
methods of delivering melphalan to a subject in need thereof, the
method comprising administering a pharmaceutical composition or
unit dosage form of the present invention to the subject in need
thereof. The methods of the present invention include parenteral
administration of the pharmaceutical compositions or unit dosage
forms of the present invention.
[0132] In some embodiments, the pharmaceutical compositions or unit
dosage forms: (or diluted forms thereof) are intravenously
administered. Intravenous administration includes, but is not
limited to, a. bolus injection, an intravenous infusion, a limb
perfusion, a normothermic isolated limb infusion, a percutaneous
hepatic perfusion, and the like, and combinations thereof.
Administering the compositions of the present invention, can also
be performed by injection and/or drip line using a cannula, a
central line, a peripherally inserted central catheter line, and
the like.
[0133] In some embodiments, a pharmaceutical composition of the
present invention is administered as an infusion for a duration of
15 minutes to 6 hours, 30 minutes to 4 hours, 45 minutes to 3
hours, 1 hour to 2 hours, about 15 minutes, about 30 minutes, about
45 minutes, about 1 hour, about 1.5 hours, about 2 hours, about 2.5
hours, about 3 hours, about 4 hours, about 5 hours, or about 6
hours.
[0134] In Some embodiments, the present invention is directed to
parenterally administering a pharmaceutical composition or unit
dosage form of the present invention to a subject for which an oral
composition of melphalan is, for one or more reasons, not
appropriate. For example, oral compositions of melphalan may not be
appropriate because a subject may be too young, unable to swallow,
undergoing surgery, incapacitated, or have a disorder that blocks
absorption of melphalan administered via the oral route. Further,
parenteral administration of the pharmaceutical compositions of the
present invention are useful for treating conditions in subject in
which a rapid increase in the in vivo concentration of melphalan is
required.
[0135] In some embodiments, the present invention directed to
methods of treating a subject suffering from a disease or disorder
amenable to treatment with melphalan, the method comprising
administering a pharmaceutical composition or unit dosage form to
the subject. As used herein the terms "treat," "treating," and
"treatment" refer to administering a composition of the present
invention prior to the onset of clinical symptoms of a disease
state/condition so as to prevent the development of any symptom, as
well as administering the composition after the onset of one or
more clinical symptoms of a disease state/condition so as to reduce
or eliminate any such symptom, aspect or characteristic of the
disease state/condition. Such treating need not be absolute to be
useful. Additionally, the terms "treat" and "treatment" refer to
both therapeutic treatment and prophylactic, maintenance, or
preventative measures, wherein the object is to prevent or slow
down (lessen) an undesired physiological condition, disorder or
disease, or obtain beneficial or desired clinical results. For
purposes of this invention, beneficial or desired clinical results
include, but are not limited to, alleviation of a symptom or a
sign; diminishment of extent of a condition, disorder or disease;
stabilization (i.e., not worsening) of the state of condition,
disorder or disease; delay in onset or slowing of condition,
disorder or disease progression; amelioration of a condition,
disorder or disease state, remission (whether partial or total),
whether detectable or undetectable; or enhancement or improvement
of condition, disorder or disease. Treatment includes eliciting a
clinically significant response, without excessive levels of side
effects. Treatment also includes prolonging survival as compared to
expected survival if not receiving treatment.
[0136] As used herein, the term "subject" refers to warm blooded
animals such as mammals, including humans and non-humans, such as,
but not limited to, domestic and farm animals, zoo animals, sports
animals, and pets (e.g., cats, dogs, mice, guinea pigs, horses,
bovine cows, and sheep). In some embodiments, a subject is a human
subject. Human subjects suitable for administering the
pharmaceutical compositions and unit dosage forms of the present
invention include, but are not limited to, pediatric, adult, and
geriatric subjects. In some embodiments of the invention, the
subject is a pediatric subject. For example, according to the U.S.
Food and Drug Administration, a "pediatric" subject is up to 21
years of age, and includes neonates (birth to about 1 month of
age), infants (about 1 month to about 2 years of age), children
(about 2 to about 12 years of age) and adolescents (about 12 to
about 21 years of age). See Guidance for industry and FDA Staff
Premarket Assessment of Pediatric Medical Devices, U.S. Dept. of
Health and Human Services, Food and Drug Administration, Center for
Devices and Radiological Health, and Center for Biologics.
Evaluation and Research (May 14, 2004). In some embodiments of the
invention, the subject is an adult. As used herein, an "adult"
subject is 18 years of age or older. In some embodiments, a subject
is an adult that is about 50 years or older. In some embodiments of
the invention, the subject is geriatric. Geriatric subjects are at
least about 65 years of age. In some embodiments, a subject is
about 70 years of age or older.
[0137] In some embodiments, the subject is a pediatric subject
suffering from a disorder such as, but not limited to, an inborn
defect, an immunodeficiency, a combined immunodeficiency, a severe
combined immunodeficiency, a congenital neutropenia with defective
stem cells, aplastic anemia, and combinations thereof.
[0138] In some embodiments, a subject is a geriatric subject
scheduled to undergo a non-myeloablative procedure.
[0139] In some embodiments, the present invention comprises a
method for treating a subject who has or is at risk for developing
a condition amenable to treatment with melphalan, the method
comprising administering an effective amount (i.e., a
therapeutically effective amount) of a composition of the invention
to the subject. Conditions amenable to treatment with melphalan
include, but are not limited to, neoplastic disorders.
[0140] In some embodiments, a therapeutically effective amount for
administering to a subject who has or is at risk for developing a
condition amenable to treatment with melphalan is 25 mg to 125 mg,
40 mg to 110 mg, 40 mg to 75 mg, 40 mg to 60 mg, about 40 mg, about
50 mg, about 60 mg, about 75 mg, or about 100 mg of melphalan as a
hydrochloride salt. The methods of the present invention also
include titrating upward or downward from an initial melphalan dose
in order to provide a therapeutically effective melphalan dosage. A
therapeutically effective dose can be administered once, twice,
thrice, four times, five times, six times, seven times, eight
times, ten times, twelve times, or more as needed.
[0141] In some embodiments, the present invention is directed to a
method of treating a disease, disorder or condition that is
therapeutically responsive to a stem cell transplantation, the
method comprising administering to a subject in need thereof a
pharmaceutical composition or unit dosage form of the present
invention followed by the subject undergoing a stern cell
transplantation.
[0142] In some embodiments, a method of the present invention
comprises administering a pharmaceutical composition or unit dosage
form (or diluted form thereof) to a subject that suffers from a
disorder selected from: myeloma, multiple myeloma, acute
myelogenous leukemia, malignant melanoma, metastatic melanoma
(e.g., metastatic ocular melanoma, metastatic cutaneous melanoma,
and the like), breast cancer, ovarian cancer, testicular cancer,
advanced prostate cancer, a myelodysplastic syndrome, a
neuroendocrine cancer (e.g., a metastatic neuroendocrine tumor, and
the like), a metastatic adenocarcinoma tumor, a hepatocellular
carcinoma, osteogenic sarcoma, polycythemia veraplasma, plasma cell
neoplasm, amyloidosis, scleromyxedema, and combinations
thereof.
[0143] In some embodiments, a method of the present invention
comprises administering a pharmaceutical composition or unit dosage
form (or diluted form thereof) to a subject for whom a stem cell
transplantation, has been indicated (e.g., a hematopoietic stem
cell transplantation). In some embodiments, a subject for whom a
stem cell transplantation has been indicated suffers from a disease
or disorder selected from a leukemia, a cancer, a non-malignant
disease, and combinations thereof. In some embodiments, a subject
for whom a stem cell transplantation has been indicated suffers
from a disease or disorder selected from myeloma, Multiple myeloma,
a lymphoma, non-Hodgkin lymphoma ("NHL"), leukemia, acute myeloid
leukemia ("AML"), Hodgkin's disease, acute lymphoblastic leukemia
("ALL"), a myelodysplastic syndrome ("MDS"), a myeloproliferative
disorder ("MPD"), chronic myelogenous leukemia ("CML"),
neuroblastoma, aplastic anemia, chronic granulocytic leukemia, a
neuroblastoma, sickle-cell, disease, osteogenic sarcoma, Ewing's
sarcoma, a desmoplastic small round cell tumor, plasma cell
neoplasm, amyloldosis, scleromyxedema, and. combinations thereof.
In some embodiments, a subject for whom a stem cell transplantation
has been indicated is a subject who would not benefit from
prolonged treatment with, or is already resistant to,
chemotherapy.
[0144] Thus, the pharmaceutical compositions and unit. dosage forms
of the present invention are useful for treatment of a condition
amenable to treatment with melphalan, as well as use for
conditioning a subject in need thereof for receiving a stem cell
transplantation.
[0145] The amount of the pharmaceutical composition that is
administered is therapeutically effective for the treatment that is
desired. For example, a therapeutically effective amount for the
treatment of multiple myeloma refers to an amount which, when
administered, diminishes one or more symptoms associated with this
disorder.
[0146] In some embodiments, the present invention is directed to a
method of conditioning a subject in order to conduct a stem cell
transplantation, the method comprising administering an effective
amount of a pharmaceutical composition or unit dosage of the
invention (e.g., intravenously) to the subject. Thus, the
pharmaceutical compositions and unit dosage forms of the present
invention are useful for treating a subject who suffers from a
condition amenable to treatment by a stem cell transplant. As used
herein, "stem cell transplantation" includes autologous and/or
allogenic transplantation procedures.
[0147] The pharmaceutical compositions of the present invention are
suitable for administering melphalan in a "high-intensity" or
myeloablative conditioning regimen in preparation for a stem cell
transplantation, or in a "reduced intensity" conditioning regimen
in preparation for a stem cell transplantation. As used herein,
"reduced intensity" conditioning refers to dosages in which a
melphalan dose of less than 150 mg/m.sup.2 is administered to a
subject at any one dose. In some embodiments, the pharmaceutical
composition of the present invention is administered to a subject
that is 50 years of age or older who suffers from a condition
amenable to treatment by a stem cell transplant.
[0148] In some embodiments, the melphalan is administered to a
subject in need of a stem cell transplantation at a dose a 50
mg/m.sup.2 to 300 mg/m.sup.2, 50 mg/m.sup.2 to 250 mg/m.sup.2, 50
mg/m.sup.2 to 225 mg/m.sup.2, 50 mg/m.sup.2 to 200 mg/m.sup.2, 50
mg/m.sup.2 to 175 mg/m.sup.2, 50 mg/m.sup.2 to 150 mg/m.sup.2, 100
mg/m.sup.2 to 300 mg/m.sup.2, 100 mg/m.sup.2 to 250 mg/m.sup.2, 100
mg.sup.2 to 225 mg/m.sup.2, 100 mg/m.sup.2 to 200 mg/m.sup.2, 100
mg/m.sup.2 to 175 mg/m.sup.2, 100 mg/m.sup.2 to 150 mg/m.sup.2, 125
mg/m.sup.2 to 300 mg/m.sup.2, 125 mg/m.sup.2 to 250 mg/m.sup.2, 125
mg/m.sup.2 to 225 mg/m.sup.2, 1.25 mg/m.sup.2 to 200 mg/m.sup.2,
150 mg/m.sup.2 to 300 mg/m.sup.2, 150 mg/m.sup.2 to 250 mg/m.sup.2,
200 mg/m.sup.2 to 300 mg/m.sup.2, 200 mg/m.sup.2 to 250 mg/m.sup.2,
about 50 mg/m.sup.2, about 100 mg/m.sup.2, about 125 mg/m.sup.2,
about 150 mg/m.sup.2, about 175 mg/m.sup.2, about 200 mg/m.sup.2,
about 250 mg/m.sup.2, or about 300 mg/m.sup.2.
[0149] In some embodiments, the administering comprises a dosage
administered at four week intervals. In some embodiments, the
dosage is administered twice, thrice, four times, five times, six
times, eight times, or ten times. For example, in some embodiments
a dose of about 100 mg/m.sup.2 is administered three times with a
four-week interval between the doses. In some embodiments, a dose
of about 200 mg/m.sup.2 is administered twice with a four-week
interval between the doses. The final dose can be followed by a
stem cell transplantation.
[0150] The pharmaceutical compositions and unit dosage forms of the
present invention can be administered alone or in conjunction with,
other medications or pharmaceutical compositions in some
embodiments, a method of the present invention comprises to a
subject a second therapeutic agent seeded from an alkylating agent
other than melphalan, a platinum compound, an antimetabolite, a
nitrosourea, a corticosteroid, a calcineurin inhibitor, a
monoclonal antibody, a polyclonal antibody, a cytotoxic antibiotic,
an interferon, an opioid, an antihistamine, a volume expander, a
pressor agent, and combinations thereof. Additional second
therapeutic agents include, but are not limited to, cisplatin,
carboplatin, doxorubicin, bortezotmib, rituximab, thalidomide,
gemeitabine, thiotepa, fludarabine, carmustine, etoposide,
cytarabine, granulocyte colony-stimulating factor (G-CSF), ADH-1,
topotecan, palifermin, prednisone, arsenic trioxide, ascorbic acid,
busulfan, cyclophosphamide, N,N',N''-triethylenethiophosphoramide,
buthionine sulfoximine, and combinations, thereof. A second,
therapeutic agent can be administered to a subject either in a
pharmaceutical composition or unit dosage form, of the present
invention that includes at least one additional therapeutic agent
(in addition to melphalan), or as a separate pharmaceutical
composition or unit dosage.
[0151] In some embodiments, the pharmaceutical compositions and/or
unit dosage forms of the present invention are administered with
other combinations of therapeutic active agents such as, but not
limited to carmustine, etoposide, and cytarabine; busulfan and
thiotepa; doxorubicin and bortezomib; arsenic trioxide and citric
acid; thalidomide and rituximab; thalidomide and prednisone; and
busulfan, fludarabine and G-CSF.
[0152] In seine embodiments, the pharmaceutical compositions and
unit dosage forms of the present invention can enhance the
bioavailability, rate of therapeutic onset, and/or therapeutic
efficacy of melphalan. Thus, the present invention is also directed
to a method of decreasing the time to therapeutic onset of
melphalan following administration thereof, the method comprising
orally or parenterally administering to a subject in need thereof a
pharmaceutical composition or unit dosage form of the present
invention. Wherein the time to therapeutic onset of melphalan
provided by the orally or parenterally administered composition or
unit dosages less than the time to therapeutic onset of melphalan
provided by an orally administered reference composition that
excludes the cyclodextrin derivative and contains an equivalent
dose of melphalan. In some embodiments, the time to therapeutic
onset of melphalan following administration of a pharmaceutical
consumption or unit dosage form of the present invention is reduced
by at least 5%, at least 10%, at least 15%, at least 20%, at least
25%, at least 30%, at least 40%, or at least 50% compared to the
time to therapeutic onset of melphalan provided by an intravenously
administered reference composition that excludes the cyclodextrin
derivative and contains an equivalent, dose of melphalan.
[0153] In some embodiments, the dissolution of melphalan from the
dosage forms of the present invention can be related to
pharmacokinetic parameters and/or the in vivo concentration of
melphalan and/or its metabolite(s). The in vivo concentration of
melphalan and its metabolite(s), as well as pharmacokinetic
parameters associated with an active form of melphalan can be
determined by, e.g., sampling the blood plasma of a subject after
administering a composition of the present invention.
Pharmacokinetic parameters that can be measured include; but are
not limited to, AUC.sub.0-t, AUC.sub.t-.infin., AUC.sub.0-.infin.,
and In(AUC.sub.LAST).
[0154] As used herein, "AUC.sub.0-t" refers to. the Area Under the
Concentration time curve (i.e., plot of plasma concentration vs.
time) after melphalan administration. The area is conveniently
determined by the "trapezoidal rule": the data points are connected
by straight line segments, perpendiculars are erected from the
abscissa to each data point, and the sum of the areas of the
triangles and trapezoids so constructed is computed.
[0155] As used herein, "AUC.sub.t-.infin." refers to the Area Under
the Concentration time curve, wherein the last concentration is
extrapolated to baseline based on the rate constant for
elimination.
[0156] As used herein, "AUC.sub.0-.infin." refers to the sum of the
Area Under the. Concentration time curves for AUC.sub.0-t. and
AUC.sub.t-.infin..
[0157] As used herein, "In(AUC.sub.LAST)" refers to the Area Under
the Concentration time curve determined by plotting plasma
concentration on a natural logarithmic scale, using the last
measured plasma concentration as the end point.
[0158] As used herein, "IntraCV" refers to an intra-assay
coefficient of variation, which is the standard deviation within a
sample set divided by the mean value of the sample set, with the
result reported as a percentage.
[0159] In some embodiments, the bioavailability of melphalan in a
human subject from a composition of the present invention is
substantially greater than that observed upon administration of an
equivalent dose of melphalan from a formulation lacking a
cyclodextrin derivative (e.g., ALKERAN.RTM. for Injection
((GlaxoSmithKline) or Melphalan HCl Injectable (Bioniche Pharma
USA)). For example, the dosage forms of the present invention can
have an AUC.sub.0-t. or AUC.sub.0-.infin., that is at least 20% or
greater, at least 25% or greater, at least 30% or greater, at least
40% or greater, at least 50% or greater, at least 60% or greater,
or at least 70% or greater than the AUC.sub.0-t or
AUC.sub.0-.infin. observed after administration of a melphalan
formulation to a subject that contains the same amount of melphalan
and lacks a cyclodextrin derivative (e.g., ALKERAN.RTM. for
Injection (GlaxoSmithKline) or Melphalan HCl Injectable (Bioniche
Pharma USA)).
[0160] In some embodiments, the bioavailability of melphalan from a
composition of the present invention is greater than that observed
upon administration of an equivalent dose of melphalan from a
formulation lacking a cyclodextrin derivative (e.g., ALKERAN.RTM.
for Injection (GlaxoSmithKline) or Melphalan HCl Injectable
(Bioniche Pharma USA)). For example, the dosage forms of the
present invention can have an AUC.sub.0-t or AUC.sub.0-.infin. that
is at least 20% or greater, at least 25% or greater, at least 30%
or greater, at least 40% or greater, at least 50% or greater, at
least 60% or greater, or at least 70% or greater than the
AUC.sub.0-t or AUC.sub.0-.infin. observed after administration of a
melphalan formulation to a subject that contains the same amount of
melphalan and lacks a cyclodextrin derivative (e.g., ALKERAN.RTM.
for Injection (GlaxoSmithKline) or Melphalan HCl Injectable
(Bioniehe Pharma USA)). In some embodiments, the AUC.sub.0-t or
AUC.sub.0-.infin. of melphalan from a composition of the present
invention is 20% to 70%, 20% to 60%, 20% to 50%, 30% to 70%, 30% to
60%, 30% to 50%, 40% to 70%, 40% to 60%, or 50% to 70% greater than
the AUC.sub.0-t or AUC.sub.0-.infin. observed upon administration
of an equivalent dose of melphalan from a formulation lacking a
cyclodextrin derivative (e.g ALKERAN.RTM. for Injection
(GlaxoSmithKline) or Melphalan HCl Injectable (Bioniche Pharma
USA)).
[0161] In some embodiments, the maximum plasma concentration
(C.sub.max) of melphalan from a composition of the present
invention, is at least 20% or greater, at least 25% or greater, at
least 30% or greater, at least 40% or greater, at least 50% or
greater, at least 60% or greater, or at least 70% or greater than a
C.sub.max observed upon administration of an equivalent dose of
melphalan from a formulation lacking a cyclodextrin derivative
(e.g., ALKERAN.RTM. for Injection (GlaxoSmithKline) or Melphalan
HCl Injectable (Bioniche Pharma USA)). In some embodiments, the
maximum plasma concentration (C.sub.max) of melphalan from a
composition of the present invention is 20% to 70%, 20% to 60%, 20%
to 50%, 30% to 70%, 30% to 60%, 30% to 50%, 40% to 70%, 40% to 60%,
or 50% to 70% greater than C.sub.max observed upon administration
of an equivalent dose of melphalan from a formulation lacking a
cyclodextrin derivative (e.g., ALKEIAN.RTM. for Injection
(GlaxoSmithKline) or Melphalan HCl injectable (Bioniche Pharma.
USA)).
[0162] In some embodiments, the rate of therapeutic onset of
melphalan from a composition of the present invention is faster
than that observed upon administration of an. equivalent dose of
melphalan from a formulation lacking a cyclodextrin derivative
(e.g., ALKERAN.RTM. for injection (GlaxoSmithKline) or Melphalan
HCl Injectable (Bioniche Pharma USA)). For example, the dosage
forms of the present invention have a time to C.sub.max (i.e.,
t.sub.max) that is. about 5%, about 10%, about 15%, about 20%,
about 25%, or about 30% faster, or about 35% faster than a
t.sub.max observed upon administration of an equivalent dose of
melphalan from a formulation lacking a cyclodextrin derivative
(e.g., ALKERAN.RTM. for Injection (GlaxoSmithKline) or Melphalan
HCl Injectable (Bioniche Pharma USA)). In some embodiments, the
dosage forms of the present invention have a time to C.sub.max
(i.e., t.sub.max) that is 5% to 35%, 5% to 30%, 5% to 25%, 5% to
20%, 10% to 35%, 15% to 35%, 20% to 35%, or 25% to 30% faster than
a t.sub.max observed upon administration of an equivalent dose of
melphalan from a formulation lacking a cyclodextrin derivative
(e.g., ALKERAN.RTM. for Injection (GlaxoSmithKline) or Melphalan
HCl Injectable (Bioniche Pharma USA)).
[0163] In some embodiments, the pharmaceutical compositions of the
present invention provide a reduced rate of hypersensitivity in
patients after parenteral administration compared with patients
parenterally administered, a similar dose of melphalan without a
cyclodextrin derivative.
[0164] In some embodiments, the pharmaceutical compositions of the
present invention provide a reduced rate of severe myelotoxicity in
patients (e.g., patients who experience a white blood cell count
<1,000 per mL and/or platelet count <25,000) after parenteral
administration compared with patients parenterally administered a
similar dose of melphalan without a cyclodextrin derivative.
[0165] In some embodiments, the pharmaceutical compositions of the
present invention provide a reduced rate of death in, patients
after parenteral administration compared with patients parenterally
administered a similar dose of melphalan without a cyclodextrin
derivative.
[0166] Raving generally described the invention; a further
understanding can be obtained by reference to the examples provided
herein. These examples are given for purposes of illustration, only
and are not intended to be limiting.
EXAMPLES
Example 1
[0167] The dissolution rate of free-base melphalan (Chemwerth,
Woodbridge, Conn.) in solutions at various pH and at various
concentrations of a cyclodextrin derivative were examined. The
procedure was as follows: free base melphalan was added to a
solution containing a cyclodextrin derivative
(SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM.) and then vortex mixed for
1-5 minutes, and, if necessary, sonicated in ice water until a
clear solution was achieved.
TABLE-US-00001 TABLE Dissolution times for free base melphalan as a
function of cyclodextrin derivative concentration, volume, and
Target Melphalan Target Melphalan SBE.sub.6.5-.beta.-CD Dissolution
Conc. Conc. Volume pH Tim (min) 50 mg/mL 200 mM 5 mL 5 50 50 mg/mL
125 mM 6 mL 5 90 50 mg/mL 100 mM 7 mL 5 160 50 mg/mL 75 mM 8 mL 5
>180 50 mg/mL 50 mM 10 mL 5 360 50 mg/mL 125 mM 6 mL 2.7 75 50
mg/mL 125 mM 10 mL 1.8 16 50 mg/mL 125 mM 6 mL 1.3 5 50 mg/mL 75,
100 125 mM 10 mL 1.1 <5 for all
[0168] Referring to the data in the above Table; the dissolution of
free base melphalan was very rapid at pH 1.1 regardless of the
concentration of the cyclodextrin derivative. After dissolution,
the solution was then neutralized with sodium hydroxide.
[0169] The solutions of free base melphalan in this example can be
prepared by addition of free base melphalan to a solution that
contains the cyclodextrin derivative, or by adding a 0.1 M HCl
solution to the melphalan and then adding the cyclodextrin
derivative, or dissolving the free base melphalan and cyclodextrin
derivative simultaneously. However, sonication was superior to
mixing and/or shaking for dissolution enhancement and de-clumping
of dry material in solution.
Example 2
[0170] The: binding of free-base: melphalan (Chemwerth, Woodbridge,
Conn.) was studied as a function of cyclodextrin derivative
concentration at pH 5 and pH 7, and the data was compared with
literature reports of free-base melphalan binding. The pH solutions
contained 100 mM sodium bitartrate buffer, and was adjusted to pH 5
using sodium hydroxide in 0.9% sodium chloride solution. The pH 7
solutions contained 50 mM each of mono- and di-basic phosphate and
0.9% sodium chloride. Solutions containing 0, 50, 75 and 100 mM
SBE.sub.6.5-.beta.-CD (CAPTISOL.RTM.) were prepared, and excess
free base melphalan was added to 2 mL samples of each solution.
After the addition of free base melphalan, the samples were vortex
mixed for 30 seconds, sonicated in ice water bath for 20 minutes,
and then mixed by end-over-end rotation at room temperature for 30
minutes. The samples were then centrifuged, the clear supernatant
was diluted with water, and analyzed by HPLC.
[0171] All melphalan assays performed by HPLC utilized the
following protocol. A Shimadzu HPLC equipped with a SCL-10A system
controller, SIL-10A auto injector, LC-10AT liquid chromatograph,
SPD-10A UV spectrophotometer detector, CTO-10A column oven, and
Class-VP chromatography laboratory automated software was utilized.
The column was a ZORBAX.RTM. RX-C18 4.6 mm by 150 mm column
(Agilent Technologies) having a 5 .mu.m particle size. Samples were
injected (20 .mu.L) onto the column for isocratic elution using
mobile phase of phosphate buffered saline (pH 7.4):methanol:glacial
acetic acid in a ratio of 500:250:10 (v/v). The mobile phase was
selected in order to decrease or nominally quench melphalan
conversion by having a high-chloride concentration. The samples
were prepared immediately prior to injection. Detection was at 260
nm.
[0172] The literature procedure involved adding an excess amount of
melphalan to 0, 10, 20, 30, 40, 50, 75 and 100 mM solutions of
SBE.sub.7-.beta.-CD (avg. M.W.=2248 g/mol) in a 25 mM phosphate
buffer solution at pH 7.5. The suspensions were placed in tightly
capped vials, sonicated 1 h, and agitated at 25.degree. C. for 23
h. The solutions were then centrifuged, the clear supernatant was
diluted with doubly distilled water, and analyzed by HPLC. See D.
Q. Ma et al., J. Pharm. Sci. 89:275 (1999);
[0173] The data from the free base melphalan solubilization studies
are provided in FIG. 1. Referring to FIG. 1, free base melphalan
displayed a significantly lower solubility enhancement than that
provided in a previous literature report. See id. Because the
solubility enhancement of free base melphalan provided by
SBE.sub.6.5-.beta.-CD was lower than expected, additional phase
solubility tests were performed using melphalan hydrochloride.
Example 3
[0174] The binding of melphalan hydrochloride (USP reference
standard) and free base melphalan (Chemwerth) with a cyclodextrin
derivative (SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM., avg. M.W.=2163
g/mol) was determined as a function of cyclodextrin derivative
concentration at pH 7.5. The temperature was maintained at
22.degree. C., and a 25 mM phosphate buffer was added to each
solution. The data was compared with a literature report of
free-base melphalan binding with SBE.sub.7-.beta.-CD (avg.,
M.W.=2248 g/mol) in a 25 mM phosphate buffer at pH 7.5 (see Example
2).
[0175] The samples were prepared by adding excess melphalan
hydrochloride, or free base melphalan to a 1 mL sample of various
SBE.sub.6.5-.beta.-CD solutions. The samples were vortex mixed for
30 seconds, sonicated at 20-24.degree. C. for 60 minutes, and then
mixed by end-over-end rotation at 22.degree. C. for 60 minutes. The
samples were then centrifuged, the clear supernatant was diluted
with water, and analyzed by HPLC. The data are provided in FIG. 2.
Referring to FIG. 2, the melphalan hydrochloride salt displayed a
significant solubility enhancement compared to free base melphalan
for all cyclodextrin derivative concentrations above 25 mM.
Example 4
[0176] A pharmaceutical composition comprising melphalan as a
hydrochloride salt was prepared by the process outlined
schematically in FIG. 3. Referring to FIG. 3, water for injection,
USP was placed in a stainless steel, mixer at a temperature of
15-20.degree. C., and hydrochloric acid was added until a pH of
about 4.6 was achieved. The resulting solution was stirred at a
speed sufficient to produce a vortex (but without foaming or
frothing) for about 15 minutes, a cyclodextrin derivative (27.2 g
SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM.) was added slowly while vortex
stirring, and the resulting solution was stirred for about 15
minutes to ensure complete dissolution. The resulting solution had
a pH of about 2.5. Melphalan as hydrochloride salt (516 mg) was
added slowly while vortex stirring, and the resulting solution was
stirred for about 15 minutes to ensure complete dissolution. A base
(2 N NaOH) was then slowly added while vortex stirring until the
solution had a pH of about 5.6. The solution, was then assayed
using a UV/vis spectrophotometer (detection wavelength of 260 nm).
The solution comprised melphalan at a concentration of 5.16 mg/ML,
and the melphalan was present in a ratio of about 1:55 w/w relative
to the cyclodextrin derivative. The solution was then passed
through a sterile filter (0.22 .mu.m PVDF) and cooled to
10-15.degree. C.
Example 5
[0177] The liquid pharmaceutical composition provided in Example 4
was lyophilized to provide a reconstitutable and/or dilutable dry
powder comprising 50 mg of melphalan as a hydrochloride salt. Glass
Vials were filled with the solution (10 mL) and placed in trays on
a pre-cooled shelf at 5.degree. C. The vials were allowed to
thermally equilibrate for about 30 minutes, and were then
lyophilized to provide a dry powder in each vial. The vials were
back-filled with nitrogen at a pressure of about 400 mTorr, and
then sealed.
Example 6
[0178] A pharmaceutical composition comprising melphalan as a
hydrochloride salt was prepared by the process described in Example
4, and outlined schematically in FIG. 3, except that the final
solution contained melphalan at a concentration of 10 mg/mL, and
the melphalan was present in a ratio of about 1:27 w/w relative to
the cyclodextrin derivative.
Example 7
[0179] The liquid pharmaceutical composition provided in Example 6
was lyophilized to provide a reconstitutable and/or dilatable dry
powder comprising 200 mg of melphalan as a hydrochloride salt.
Glass vials were filled with the solution (20 mL) and placed in
trays on a pre-cooled Shelf at 5.degree. C. The vials were allowed
to thermally equilibrate for about 30 minutes, and were then
lyophilized to provide a dry powder in each vial. The vials were
back-filled with nitrogen at a pressure of about 400 mTorr, and
then sealed, packaged, and labeled. The vials were protected from
exposure to light during all aspects of the lyophilization,
back-filling, sealing, packaging and labeling procedures.
Prophetic Example A
[0180] The liquid pharmaceutical composition provided in Examples 4
and 6 will be aseptically spray dried to provide a free-flowing
powder to be filled aseptically. The free-flowing powder will meet
or exceed the dissolution properties of the lyophilized powder
prepared in Examples 4 or 6.
Example 8
[0181] A pharmaceutical composition comprising melphalan as a
hydrochloride salt was prepared by the process outlined
schematically in FIG. 4. Referring to FIG. 4, water for injection,
USP was placed in a stainless steel mixer at a temperature of
18-25.degree. C., and the resulting solution was stirred at a speed
sufficient to: produce a vortex (but without foaming or frothing).
A cyclodextrin derivative (SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM.)
was added slowly while vortex stirring, and the resulting solution
was stirred for about 15 minutes to ensure complete dissolution.
The resulting solution was then cooled to about 2-8.degree. C.
Melphalan as a hydrochloride salt was added slowly while vortex
stirring, and the resulting solution was stirred for about 15
minutes to ensure complete dissolution. A base (2 N NaOH) was then
slowly added while vertex stirring until the solution had a pH of
about 5-6 (target pH 5.5). An in-process control ("IPC") assay was
then, performed to monitor pH, and the solution was diluted to the
final target volume using water for injection, USP. The solution
was then assayed using a UV/vis spectrophotometer (detection
wavelength, of 260 nm) and a bioburden assay was performed. The
solution was then passed through a sterile filter (0.22 .mu.m PVDF)
and cooled. to 15-25.degree. C. Finally, an IPC assay was
conducted.
Example 9
[0182] The solution prepared in Example 8 was lyophilized to
provide a reconstitutable and/or dilutable dry powder comprising
melphalan as a hydrochloride salt. For the lyophilization, glass
vials were filled with the solution (10 mL) and placed in trays on
a pre-cooled shelf at. 5.degree. C. The vials were allowed to
thermally equilibrate for about 1 hour, and were lyophilized to
provide a dry powder in each vial. The vials were back-filled with
nitrogen, sealed, packaged, and labeled. The vials were protected
from exposure to light during all aspects of the lyophilization,
back-filling, sealing, packaging and labeling procedures.
Prophetic Example B
[0183] The liquid pharmaceutical composition provided in Example 8
will be aseptically spray dried to provide a free-flowing powder to
be filled aseptically. The free-flowing powder will meet or exceed
the dissolution properties of the lyophilized powder prepared in
Example 9.
Example 10
[0184] The properties of the pharmaceutical compositions of the
present invention after dilution with Water for Injection, USP,
were analyzed by a variety of analytical methods. The results are
listed in the Table below. Compositions A-D were prepared by the
process described in Examples 8-9. The diluted compositions
contained SBE.sub.6.5-.beta.-CD (CAPTISOL.RTM.) in a concentration
of 75 mM, 100 mM, 125 mM, and 125 mM, respectively. Each of the
compositions had a moisture content of about 1.3% to about 2.5%
prior to dilution.
TABLE-US-00002 TABLE Properties of pharmaceutical compositions of
the present invention containing varying concentrations of a
cyclodextrin derivative. Dilution SBE.sub.6.5-.beta.-CD Dissolution
Density Time for Volume Conc. Time pH (22.degree. C.) Viscosity 5%
loss A 10 mL 75 mM <30 s 5.05 1.07 g/cc 2.06 cP 10 h B 10 mL 100
mM <30 s 4.9 1.08 g/cc 2.28 cP 23 h C 10 mL 125 mM 45 s 5.05
1.11 g/cc 2.95 cP 49 h D 5 mL 125 mM 105 s 5.2 1.11 g/cc 3.02 cP 25
h
Example 11
[0185] The stability of melphalan hydrochloride upon dilution of a.
pharmaceutical composition of the present invention was determined.
Pharmaceutical compositions containing a cyclodextrin derivative
(SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM., avg. M.W.=2163 g/mol) were
diluted with isotonic saline to provide 0.45 mg/mL melphalan
solutions that contained the cyclodextrin derivative at a
concentration of 75 mM and 125 mM, respectively. Melphalan was
assayed as a function of time, and the time necessary for a 5% or
10% loss of melphalan (based on an initial melphalan concentration
of 100%) was determined. The data is provided in the Table below.
The times required for melphalan to fall to 90% or 95% of its
initial concentration in the solutions that contained the
cyclodextrin derivative were compared to the stability of melphalan
in a Reference product (ALKERANR for Injection,
GlaxoSmithKline).
TABLE-US-00003 TABLE Melphalan stability as a function of
cyclodextrin derivative concentration, compared to a reference
melphalan standard. Time for 5% loss Time for 10% loss
SBE6.5-.beta.-CD (75 mM) 5.4 h 11 h SBE6.5-.beta.-CD (125 mM) 8.8 h
18 h Reference 1.3 h 2.7 h
[0186] Referring to the data in the above Table, the stability of
melphalan after dilution from a pharmaceutical composition of the
present invention shows an improvement of approximately 4.2 times
and 6.8 times at a cyclodextrin derivative concentration of 75 mM
and 125 mM, respectively, compared to a reference melphalan
formulation that does not contain a cyclodextrin derivative.
Example 12
[0187] The stability of melphalan hydrochloride upon dilution of a
pharmaceutical composition of the present invention was determined
as a function of temperature and storage conditions. Pharmaceutical
compositions containing melphalan (50 mg) and a cyclodextrin
derivative (SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM., avg. M.W.=2163
g/mol, 270 mg) were diluted with isotonic saline (8.5 mL) to
provide a concentrated solution. The concentrated solution was
further diluted 10-fold to provided a dilute solution. Each of the
concentrated and diluted melphalan solutions were stored at
25.degree. C./60. relative humidity, or in a refrigerator
(.about.10.degree. C.), and the melphalan content was monitored as
a function of time. The data is provided in the Table below.
TABLE-US-00004 TABLE Melphalan stability as a function of
temperature and storage conditions. Storage Melphalan Monohydroxy
Solution Time Conditions Assay Melphalan Assay Conc. Sol'n 0
Refrigerator 99% 0.8% '' 6.5 '' 98.9%.sup. 0.8% '' 24.5 ''
98.8%.sup. 0.9% '' 48.5 '' 98.4%.sup. .sup. 1% Conc. Sol'n 0
25.degree. C./60% R.H. 99% 0.8% '' 6 '' 98% 1.5% '' 24 '' 96% 3.4%
'' 48 '' 93% 5.7% Dilute Sol'n 0 Refirgerator 99% .sup. 1% '' 5 ''
98.4%.sup. 1.4% '' 24.3 '' 97.8%.sup. .sup. 2% '' 48.4 ''
96.7%.sup. 2.7% Dilute Sol'n 0 25.degree. C./60% R.H. 99% .sup. 1%
'' 5.3 '' 94% 5.3% '' 23.8 '' 81% 15.5% '' 47.2 '' 70% 20.2%
[0188] Referring to the data in the above Table, the stability of
melphalan after dilution from a pharmaceutical composition of the
present invention provides a significant improvement compared to
currently available melphalan pharmaceutical compositions that do
not contain a cyclodextrin derivative.
Example 13
[0189] The stability of a lyophilized melphalan hydrochloride
composition was determined before and after dilution as a function.
of temperature and storage conditions. The study was performed
under the direction of CyDex Pharmaceuticals, Inc. by
BioConvergence LLC, Bloomington,
[0190] Compositions comprising melphalan (50 mg) and povidone (20
mg) were diluted with compositions comprising a cyclodextrin
derivative (SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM., avg. M.W.=2163
g/mol, 270 mg), sodium, citrate (200 mg), and distilled water (10
mL) to provide concentrated melphalan solutions (5 mg/mL). In
addition to testing the stability of the concentrated solutions,
further dilution 11-fold to provide dilute solutions containing
melphalan (0.45 mg/mL).
[0191] The kinetic stability of a reconstituted concentrated
solution (5 mg/mL melphalan) was determined upon storage in a glass
vial, and the kinetic stability of a reconstituted diluted solution
(0.45 mg/mL melphalan) was determined upon storage in a 50 mL
Baxter INTRAVIA.RTM. bag, at refrigerated (about
2.degree.-8.degree. C.) and room (about 25.degree. C., monitored
under fluorescent light) temperatures.
[0192] Evaluation of the kinetic stability of the dilute (0.45
mg/ml melphalan) composition was determined in a 50 mL Type I glass
container: reconstitution was performed using saline (8.5 mL), and
aliquots (4.5 mL) were removed from each vial and injected into 4
glass containers that held 45.5 mL of saline. After an amount was
withdrawn from each container for t=0 analysis, the containers were
stored at room temperature ("RT", about 25.degree. C., under
fluorescent light), or in a refrigerator (about 2.degree.-8.degree.
C.), and the melphalan content was monitored as a function of time.
The data is provided in the Table below.
TABLE-US-00005 TABLE Summary of melphalan stability as a function
of temperature and storage conditions MEL Hold Degradation Run Form
Storage Conditions Time (% w/w) 1 Lyophilized comp. RT
(~20.degree.-25.degree. C.) 2 yrs <2% Conc. Solution (5 mg/mL)
Immediately diluted n/a n/a Dilute solution (0.45 mg/mL) RT
(~20.degree.-25.degree. C.) 10 h <4% 2 Lyophilized comp. RT
(~20.degree.-25.degree. C.) 2 yrs <2% Conc. Solution (5 mg/mL)
RT (~20.degree.-25.degree. C.) 24 h <4% Dilute solution (0.45
mg/mL) RT (~20.degree.-25.degree. C.) 5 h <2% 3 Lyophilized
comp. RT (~20.degree.-25.degree. C.) 2 yrs <2% Conc. Solution (5
mg/mL) Refrigerated (2.degree.-8.degree. C.) 48 h <4% Dilute
solution (0.45 mg/mL) RT (~20.degree.-25.degree. C.) 5 h <2% 4
Lyophilized comp. RT (~20.degree.-25.degree. C.) 2 yrs <2% Conc.
Solution (5 mg/mL) immediately diluted n/a n/a Dilute solution
(0.45 mg/mL) 1) Refrigerated (2.degree.-8.degree. C.) 1) 24 h 1)
<2% .sup. 2) RT (~20.degree.-25.degree. C.) 2) 5 h 2) <2%
.sup.
[0193] Referring to the data in the above Table, the stability of
melphalan after dilution from a pharmaceutical composition of the
present invention provides a stable composition that can be
maintained at room temperature for up to 5 hours and exhibit less
than 2% melphalan degradation, or up to 10 hours and exhibit less
than 4% melphalan degradation. When refrigerated, a diluted
melphalan composition can be stored up to 24 hours and exhibit less
than 2% melphalan degradation. Additionally, a lyophilized
pharmaceutical composition can be stored up to 2 years at room
temperature and exhibit less than 2% melphalan degradation.
[0194] Furthermore, referring to Examples 12 and 13, the
pharmaceutical compositions of the present invention provide a
significant improvement in melphalan stability compared to other
formulations in which the use of a cyclodextrin derivative has been
proposed. For example, D. Q. Ma et al., Int. J. Pharm. 189:227
(1999) provide a melphalan composition that upon dilution with a
solution containing a cyclodextrin derivative, exhibits a melphalan
loss of more than 60% after 48 hours at room temperature.
Significantly, the data in the above Tables illustrates that upon
dilution of a pharmaceutical composition of the present invention,
a melphalan loss of at most 30% is observed within 45 hours at room
temperature. The melphalan loss can be reduced to as low as 2% or
3% when the solution is stored at a reduced temperature (e.g., in a
refrigerator).
Example 14
[0195] The hemolytic potential of a cyclodextrin derivative
suitable for use with the pharmaceutical composition of the present
invention was analyzed in comparison to a previously marketed
diluent vehicle for melphalan (ALKERAN.RTM. For injection,
GlaxoSmithKline). The hemolytic potential was evaluated in rodent
(SPRAGUE DAWLEY.RTM. or Wistar Han IGS rats) and human red blood
cells obtained from fasted subjects using a spectrophotometric
technique. Normal saline (0.9% sodium chloride) was used as the
blank (or background) and as a negative control for comparison
against various concentrations of a cyclodextrin
derivative-containing, and cyclodextrin-free diluent vehicles. A
positive control containing Triton X-100 (1%) in phosphate buffered
saline was also utilized. Human red blood cells were taken from
fasted (.gtoreq.8 h) adult subjects. The components of the various
samples are listed in the following
TABLE-US-00006 TABLE Components of diluent vehicles used for
hemolysis studies. Identification Constituents Negative
Control/Bank 0.9% Sodium Chloride Positive Control 1% TRITON .RTM.
X-100 in phosphate buffered saline Cyclodextrin Derivative
SBE.sub.6.5-.beta.-CD (CAPTISOL .RTM., 9.72 g) Diluent q.s. 400 mL
with normal saline ALKERAN .RTM. for Injection Povidone (K-12, 72
mg) (GlaxoSmithKline) Diluent Sodium citrate (720 mg) Propylene
glycol (21.6 mL, 22.4 g) Ethanol (1.87 mL, 1.48 g) Water (12.2 mL)
q.s. 400 mL with normal saline
[0196] The rat and human red blood cells were exposed to various
concentrations of the diluent vehicles and the hemolytic potential
was evaluated using. equation (1):
A.sub.541(test article)-A.sub.540(negative control).times.100=%
Hemolysis (1)
[0197] The hemolysis results are provided in the following Table,
where Group A refers to rat red blood dells exposed to the
cyclodextrin derivative vehicle; Group B refers to rat red blood
cells exposed to the ALKERAN.RTM. for injection (GlaxoSmithKline)
diluent vehicle; Group C refers to human red blood cells exposed to
the cyclodextrin derivative vehicle; and Group D refers to human
red blood cells exposed to the ALKERAN.RTM. for Injection
(GlaxoSmithKline) diluent vehicle. The negative controls for each
experiment provided absorbances below 0.13, and the positive
controls for each experiment provided absorbances of about 2.8 to
3.
TABLE-US-00007 TABLE Hemolysis results for rat and human red blood
cells exposed to various diluent vehicles. Dilution Group Meas.
none 1:2 1:4 1:8 1:16 1:32 1:64 1:128 A Abs. (a.u.) 0.11 0.114 0.11
0.117 0.116 0.118 0.119 0.118 Hem. (%) 1% 0 0 0 0 0 0 0 B Abs.
(a.u.) 0.178 0.155 0.141 0.128 0.125 0.121 0.123 0.124 Hem. (%) 2%
1% 1% 0 0 0 0 0 C Abs. (a.u.) 0.021 0.021 0.022 0.019 0.017 0.016
0.017 0.025 Hem. (%) 0 0 0 0 0 0 0 0 D Abs. (a.u.) 0.037 0.031
0.033 0.02 0.025 0.022 0.027 0.027 Hem. (%) 0 0 0 0 0 0 0 0
[0198] Referring to the hemolysis data in the above Table, the
Solution that contained the cyclodextrin: derivative at high
concentrations (e.g., no dilution, 1:2 dilution' with saline, and
1:4 dilution with saline) provided reduced hemolysis in rat red
blood cells, which was also exhibited as reduction in
spectrophotometric absorption of about 30% compared to the
ALKERAN.RTM. for Injection (GlaxoSmithKline) diluent. While, the
hemolysis tests in human blood cells exhibited, a similar reduction
in spectiophototnetrie absorbance at high concentrations, neither
the cyclodextrin derivative solution or the ALKERAN.RTM. for
Injection (GlaxoSmithKline) diluent vehicle induced homolyses in
human red blood, cells.
Example 15
[0199] A study was conducted that determined melphalan associated
with SAE.sub.6.5-.beta.-CD (CAPTISOL.RTM., CyDex Pharmaceuticals,
Inc., Lenexa, Kans.) exhibits the same protein binding as
unassociated melphalan. The study was performed under the direction
of CyDex Pharmaceuticals, Inc. by Analytical Biochemistry
Laboratories, Inc., Columbia, Mo.
[0200] Preliminary Study
[0201] A preliminary study was performed that determined
radioactive-labeled melphalan, [.sup.14C]-melphalan (Moravek
Biochemicals, Inc., Brea, Calif.), does not bind non-specifically
to ultrafiltration devices. The following mixtures of compounds
were added to human plasma ultrafiltrate (Biochemed, Winchester,
Va.) to determine the protein binding of [.sup.14C]-melphalan alone
or in combination with SAE.sub.6.5-.beta.-CD: [0202] 1.
[.sup.14C]-melphalan with melphalan; and [0203] 2.
[.sup.14C]-melphalan with melphalan and SAE.sub.6.5-.beta.-CD.
[0204] Radioactive-labeled warfarin, [.sup.3H]-warfarin (Moravek
Biochemicals, Inc., Brea, Calif.), a compound with well-documented
protein binding properties was used as a positive control in all
experiments.
[0205] Powdered material (as applicable) was weighed into
scintillation vials (20 mL) and the radiolabeled compounds were
added to the vials using a positive displacement pipette. Blank
human plasma ultrafiltrate (5 mL) was then added to the vials using
a glass serological pipette. The mixtures were then blended
briefly. The time-dependence of plasma protein binding was
determined by sampling the mixtures 0.5, 1, and 5 minutes after
addition of the test compounds to the human plasma ultrafiltrate.
The sample aliquots (3.times.1 mL) were dispensed into the
ultrafiltration devices, and the samples were immediately
centrifuged (1600 g for 5 minutes at 25.degree. C.). The solution
remaining in the vials was then aliquoted for Liquid Scintillation
Counting (LSC) analysis (2.times.0.1 mL).
[0206] More than 95% recovery was observed for [.sup.3H]-warfarin
in all experiments. In the protein binding experiments,
[3H]-warfarin was over 99% protein bound. Radiolabeled melphalan
(alone or in combination with. SAE.sub.6.5-.beta.-CD) applied to
ultrafiltration devices having a molecular weight cutoff of 30 kD
exhibited an average of over 97% radioactivity recovery:
[.sup.14C]-melphalan alone exhibited a recovery of 97.7% (n=3), and
[.sup.14C]-melphalan with SAE.sub.6.5-.beta.-CD exhibited a
recovery of 97.6% (n=3). The results demonstrate that there was
minimal (i.e., less. than 2.4%) non-specific binding of
radiolabeled melphalan to the ultrafiltration devices.
[0207] Protein-Binding Study
[0208] For the protein-binding study, the radiolabeled melphalan,
non-radiolabeled melphalan, and SAE.sub.6.5-.beta.-CD (as
applicable) were added to a scintillation vial (20 mL) and blown to
dryness under a nitrogen stream to standardize the amount of
solvent present in each experiment. Methanol (50 .mu.L) was added
to the vials and blank human plasma (5 mL) was added to the vials
using a glass serological pipette. The mixtures were then blended
briefly. Aliquots (3.times.1 mL) were then dispensed from the vials
into the ultrafiltration devices, followed by centrifugation (2,000
g for 5 minutes at 25.degree. C.). The time interval between adding
human plasma to the vials and the start of centrifugation was 0.5,
1, 5, 10, and 30 minutes. The solution remaining, in the vials was
then aliquoted in duplicate (at a volume of 0.1, 0.05, and 0.025
mL) for LSC analysis.
[0209] Sample radioactivity was quantified using a scintillation
counter (Beckman Instruments, Inc. Schaumberg, Ill.) equipped with
the H-number method for cpm to dpm conversion. LSC analysis was
performed with samples (5 mL) in glass scintillation vials (7 mL),
from which background measurements were made using the same amount
of scintillation fluid added to the vials. The results are provided
in the following Table:
TABLE-US-00008 TABLE Protein binding of radiolabeled melphalan
("[.sup.14C]-mel") with unlabeled melphalan ("mel") in the presence
and absence of SAE.sub.6.5-.beta.-CD. Total Conc. Conc. Recovery
(%) % Binding Mixture (dpm/mL) (.mu.g/mL) Rep. Data Avg. Data Avg.
[.sup.14C]-mel + mel 112,105 14 1 96.4 97.1 64.5 64.3 .+-. 0.34 (1
min.) 2 96.9 63.9 3 98.0 64.4 [.sup.14C]-mel + mel + 112,382 14 1
92.9 91.5 63.7 64.3 .+-. 0.69 SAE.sub.6.5-.beta.-CD 2 95.4 64.1
(0.5 min.) 3 86.4 65.1 [.sup.14C]-mel + mel + 108,085 13 1 97.3
97.5 63.8 64.4 .+-. 0.59 SAE.sub.6.5-.beta.-CD 2 97.0 64.8 (1 min.)
3 98.3 64.6 [.sup.14C]-mel + mel + 109,621 13 1 96.5 95.9 65.8 67.2
.+-. 1.96 SAE.sub.6.5-.beta.-CD 2 95.7 69.5 (5 min.) 3 96.5 66.4
[.sup.3H]-warfarin 114,932 0.052 1 98.5 101 98.9 98.8 .+-. 0.01 2
100 98.8 3 103 98.8
[0210] The results showed that [.sup.14C]-melphalan in the absence
of SAE.sub.6.5-.beta.-CD was 64.3% protein bound after 1 minute in
human plasma. Similar degrees of protein binding were observed for
[.sup.14C]-melphalan in the presence of SAE.sub.6.5-.beta.-CD,
64.3% (0.5 minutes, n=3), 64.4% (1 minute, n=3), and 67.2% (5
minutes, n=3). The study showed that SAE.sub.6.5-.beta.-CD did not
affect the protein-binding of radiolabeled [.sup.14C]
melphalan.
Example 16
[0211] A study was performed to investigate the potential for a
sulfoalkyl either cyclodextrin to perturb the in vivo
pharmacokinetics of melphalan. Pharmacokinetic parameters were
determined for melphalan following intravenous administration to
male Sprague Dawley rats in the presence or absence of
SBE.sub.6.5-.beta.-CD in the delivery vehicle.
[0212] The pharmacokinetics of melphalan were studied in
overnight-fasted male Sprague Dawley rats. All experimental
procedures were approved, and performed in accordance with the
guidelines of the Institutional Animal Experimentation Ethics
Committee (Monash University Ethics approval number
VCPA/2008/02).
[0213] On the day prior to dosing, a commercially available BASi
CULEX.RTM. cannula (for use with a CULEX.RTM. automated blood
sampling device) was inserted into the left carotid artery of each
rat under isoflurane anesthesia (2%). A polyethylene cannula was
also inserted into the right jugular vein. Cannulae were
exteriorized by tunneling subcutaneously to emerge above the
scapulae.
[0214] Immediately following surgery and through to the end of the
experiment, rats were housed in RATURN.RTM. metabolic cages in the
CULEX.RTM. automated blood sampler. All rats returned to normal
grooming, drinking and sleeping behavior within an hour of surgery.
Animals were given a small amount of food just after they awoke
from the anesthetic, but were then fasted for 16-18 hours prior to
drug administration. Animals had access to water ad libitum at all
times. Food was reinstated 4 hours following drug administration.
At the conclusion of each experiment, rats were killed by a single
lethal injection of pentobarbitone.
[0215] The cyclodextrin derivative-free melphalan formulation was
prepared as per the product insert for ALKERAN.RTM. for Injection
(GlaxoSmithKline). The contents of a single ALKERAN.RTM. for
Injection (GlaxoSmithKline) vial were reconstituted with 10 mL of
sterile diluent (provided with the ALKERAN.RTM. product, and
containing 0.2 g sodium citrate, 6 mL propylene glycol, 0.52 mL
ethanol (96%) and water). The solution was then further diluted
with 0.9% normal saline (2 mL ALKERAN.RTM. for Injection
(GlaxoSmithKline) in 10 mL of 0.9% saline, i.e., 12 mL total
volume), and the resultant formulation was sterilized by filtering
through a 0.22 .mu.m syringe filter before administering to rats.
The measured concentration of melphalan in the IV formulation was
0.54 mg/mL (as free base) and the pH of the final solution was
between 5 and 6 (checked using pH paper). The formulation was
administered to animals within 30 minutes of preparation.
[0216] A formulation containing SBE.sub.6.5-.beta.-CD (27% w/v) was
prepared by dissolving SBE.sub.6.5-.beta.-CD in Milli-Q water.
Contents of an ALKERAN.RTM. for Injection (GlaxoSmithKline) vial
were then reconstituted with 10 mL of the SBE.sub.6.5-.beta.-CD
solution. This solution was then diluted with 0.9% saline (2 mL in
10 mL of 0.9% saline) and the resultant formulation was sterilized
by filtering through a 0.22 .mu.m syringe filter before
administering to rats. Thus, the final formulation contained. 4.5%
(w/v) SBE.sub.6.5-.beta.-CD. The measured concentration of
melphalan (as free base) was 0.58 mg/ml, and the pH of the final
solution was between 5 and 6 (checked using pH paper). The
formulation was administered to animals within 30 minutes of
preparation.
[0217] The total dose volume was 1 mL and all doses were infused
manually via the jugular vein cannula. The complete dose was
delivered over a 10-minute period and the cannula was flushed with
heparinized saline (10 U/mL) to ensure complete administration of
the dose.
[0218] Samples of arterial blood and urine were collected
according: to the following schedules: blood/plasma sampling times
were pre-dose, and 5, 10 (end of infusion), 15, 25, 40, 55, 70,
100, 130, 190, 250, 370, and 490 minutes post-dose; and urine
sampling intervals were 0-70 minutes, 70-130 minutes, 130-190
minutes, 190-250 minutes, 250-310 minutes, 310-370 minutes, 370-430
minutes, 430-490 minutes, and 490-1450 minutes.
[0219] Arterial blood was collected directly into borosilicate
vials (at 4.degree. C.) containing heparin, COMPLETE.RTM. (a
protease inhibitor cocktail), potassium fluoride, and EDTA to
minimize potential for ex vivo degradation in blood/plasma samples.
Once collected, an aliquot (50 .mu.L) of whole blood was
transferred into a fresh micro-centrifuge tube. The remaining blood
was centrifuged and supernatant plasma removed. All blood, plasma
and urine samples were immediately (snap) frozen on dry ice and
then transferred to -20.degree. C. freezer for storage until
analysis.
[0220] The melphalan concentration in whole blood, plasma, urine
and samples of dosing solutions were determined using LC-MS.
[0221] Sample preparation was performed using protein precipitation
with acetonitrile. Aliquots of the plasma and blood (50 .mu.L) were
treated with internal standard (10 .mu.L), acetonitrile (130
.mu.L), vortexed and centrifuged. The supernatant was removed and
analyzed by LC/MS. Standard samples were prepared by spiking
solution standards in the respective blank matrix. A stock solution
of melphalan free base was prepared at a concentration of 10 mg/mL
in dimethylsulfoxide. This stock solution was further diluted in
aqueous acetonitrile (50% v/v) to obtain spiking solutions for the
preparation of calibration standards.
[0222] All samples were assayed via LC-MS/MS on a Micromass Quattro
Premier PR triple quadrupole instrument coupled with a Waters
Acquity UPLC (Waters Corp., Milford, Mass.). Analytical separations
were performed on a Phenomenex Polar reverse-phase column (50
mm.sup.x1.0 mm inner diameter, 4 .mu.m particle size) equipped with
a Phenomenex Polar Security Guard column of the same material
(Torrance, Calif.). Samples (7.5 .mu.L) were injected onto the
column and compounds were eluted (at a flow rate of 0.15 mL/min)
using a ternary gradient solvent system consisting of an aqueous
solution of methanol (2% v/v) and formic acid (0.05% v/v) Milli-Q
water (solvent A), and acetonitrile containing formic acid (0.05%
v/v) (solvent B). The gradient conditions used for LC-MS analysis
are listed in the following table.
TABLE-US-00009 TABLE Gradient chromatography conditions used for
the analysis of melphalan. Time (minutes) Solvent A Solvent B .sup.
0-0.2 89 2 0.3 80 20 2.7 20 80 2.8 5 95 3.3 5 95 3.5-4.5 98 2
[0223] Elution of analytes was confirmed by multiple reaction
monitoring (MRM) using diazepam (0.2 .mu.g/mL) as the internal
standard (diazepam). Inlet cone voltages were 20 eV and 40 eV for
melphalan and the internal standard, respectively, and collision
energies of 15 eV and 27 eV for melphalan and the internal
standard, respectively. The elution of melphalan and internal
standard was monitored using the following transitions.
304,94>26788 and 285.17>154.02, respectively. Melphalan
exhibited a retention time of 2.0 minutes and the internal standard
exhibited a retention time of 2.8 minutes,
[0224] Mass spectrometry was performed using positive mode
electrospray ionization with a capillary voltage a 3.2 kV, detector
Multiplier gain of 650 V, and source block and desolvation
temperatures of 90.degree. C. and 300.degree. C., respectively. A
desolvation gas (nitrogen) and collision gas (argon) flow of 500
L/h and 0.38. mL/min, respectively, was maintained. The lower limit
or quantitation (LLQ) for blood and plasma standards was 5.0 ng/mL,
and the LLQ for diluted urine samples was 0.5 ng/mL.
[0225] Both plasma and blood concentration data were analysed to
obtain pharmacokinetic parameters using WINNONLIN.RTM. software
WINNONLIN.RTM. professional version 5,2,1, Pharsight Corp.,
Mountain View, Calif.). The total clearance (CL.sub.total, for
whole blood or plasma) after intravenous administration was
calculated as: CL.sub.total=Dose/AUC, where AUC is the area under
the whole blood or plasma concentration versus time curve obtained
using the linear trapezoidal method. The volume of distribution
(V.sub.z) was calculated as V.sub.z=/.lamda..sub.z, where
CL.sub.total/.lamda..sub.z, where .lamda..sub.z is the elimination
rate constant after i.v. administration.
[0226] The mean dose-normalized concentration versus time profiles
of melphalan in whole blood and plasma following intravenous
administration using a formulation containing SBE.sub.6.5-.beta.-CD
(n=4) and a cyclodextrin-free (n=5) formulation are presented in
the following table.
TABLE-US-00010 TABLE Pharmacokinetic parameters for melphalan in
whole blood and plasma following intravenous administration to male
Sprague Pawley rats at a nominal dose of 2.0 mg/kg with a
formulation containing a cyclodextrin derivative
(SBE.sub.6.5-.beta.-CD) and a formulation free from a cyclodextrin
derivative ("CD-free formulation"). Whole-Blood. Plasma
SBE.6.5-1.cndot.CP CD-free SBE.sub.6.5-.beta.-CD CD-free Melphalan
(27% w/v) formulation (27% w/v) formulation Apparent t.sub.1/2 (h)
0.8 .+-. 0.1 0.9 .+-. 0.1 0.8 .+-. 0.1 0.9 .+-. 0.1 CL.sub.total
(mL/min/kg) 10.9 .+-. 2.0 12.2 .+-. 1.3 8.1 .+-. 1.1 9.3 .+-. 1.4
V.sub.z (L/kg) 0.8 .+-. 0.2 1.0 .+-. 02 0.6 .+-. 0.1 0.7 .+-. 0.2
AUC.sub.0-inf/D 94.2 .+-. 16.6 82.8 .+-. 8.8 125.4 .+-. 15.0 109.6
.+-. 16.1 (.mu.M min kg/.mu.mol)
[0227] FIGS. 5A-5B provide graphic representations of the
dose-normalized whole blood (FIG. 5A) and plasma (FIG. 5B)
concentrations of melphalan following intravenous administration
for the SBE.sub.6.5-.beta.-CD-containing: 'melphalan formulation
(.circle-solid.), n=4, and the cyclodextrin-free (o), n=5,
melphalan formulation (ALKERAN.RTM. for Injection,
GlaxoSmithKline). Referring to FIGS. 5A-5B, the data is presented
as mean with error bars indicating a. single standard deviation.
Melphalan exhibited bi-exponential pharmacokinetics it both whole
blood and plasma and the apparent terminal elimination phase was
well-defined within the: 8 hour post-dose blood sampling period,
with the apparent terminal elimination half-life. The mean whole
blood and plasma concentration versus time profiles for the
formulation that: contained SBE.sub.6.5-.beta.-CD. (27% w/V) and
cyclodextrin-free formulation were essentially super-imposable and
there were: no statistically significant differences in any of the
pharmacokinetic parameters between the two formulations
(p>0.05), Thus, as shown in the above table, -in the rat the in
vivo pharmacokinetic parameters for a melphalan formulation
containing SBE.sub.6.5-.beta.-CD are essentially identical to the
pharmacokinetic parameters for a cyclodextrin-free formulation
(i.e., ALKERAN.RTM. for Injection, GlaxoSmithKline).
[0228] Additionally, in both formulations the percentage of the
melphalan dose excreted in urine excreted as unchanged compound up
to 24 hours post-dose was low: for the cyclodextrin-free
formulation the average was 2.7.+-.1.7%, and for the formulation
containing SBE.sub.6.5-.beta.-CD the average was 2:3.+-.2%.
[0229] The data show that pharmacokinetic parameters, including
half-life, AUC, volume of distribution, clearance; and the extent
of renal elimination of melphalan were essentially unchanged
between ALKERAN.RTM. for Injection (GlaxoSmithKline) and the
melphalan formulation that contained SAE.sub.6.5-.beta.-CD/.
Specifically, the mean whole blood and plasma concentration versus
time profile of melphalan with and without SBE.sub.6.5-.beta.-CD
are essentially super-imposable. The results demonstrate that
SBE.sub.6.5-.beta.-CD had no observable difference in, blood or
plasma versus time profiles for melphalan in the rat model.
Furthermore, there was no apparent difference in the urinary
excretion of melphalan: in the rat model.
Example 17
[0230] A Phase lla, multicenter, open-label, randomized, efficacy
and safety study of melphalan hydrochloride administered by
injection using a propylene glycol-free diluent vehicle has been
conducted in 3 human multiple myeloma patients who underwent
myeloablative conditioning in preparation for autologous
transplantation. The study is on-going.
[0231] The primary goal of the study was/is to determine the rate
of myeloablation and neutrophil engraftment in multiple myeloma
patients who receive a high-dose of melphalan hydrochloride via
injection in which one dose is administered using a propylene
glycol diluent and one: dose is administered using a propylene
glycol-free diluent. The administering, is used as myeloablative
therapy prior to autologous stem cell transplantation.
[0232] The secondary goal of the study is to determine: (a) the
rate of platelet engraftment in multiple myeloma patients receiving
high-dose melphalan hydrochloride via injection using both a
propylene glycol diluent and, a propylene glycol-free diluent prior
to autologous: stem cell transplantation; (b) the median, time to
engraftment of neutrophil and platelets in multiple myeloma
patients receiving high-dose melphalan hydrochloride via:
injection, using both a propylene glycol diluent and a propylene
glycol-free diluent prior to autologous stem cell transplantation;
(c) the response rate (stringent complete response [sCR], complete
response [CR], very good partial response [VGPR], partial response
[PR], stable disease [SD], or progressive disease [PD]) at
autologous stem cell transplantation at day +100 in multiple
myeloma patients receiving high-dose melphalan hydrochloride via
injection using a propylene glycol diluent and a propylene
glycol-free diluent, prior to autologous stem cell transplantation;
(d) the toxicity profile of high-dose melphalan hydrochloride via
injection using both a propylene glycol diluent and a propylene
glycol-free diluent in multiple myeloma patients undergoing
autologous stem cell transplantation; (e) the rate of
treatment-related mortality during the first 100 days after
autologous stem cell transplantation in multiple myeloma patients
receiving high-dose melphalan hydrochloride via injection, using a
propylene glycol diluent and a propylene glycol-free diluent; and
(f) the pharmacokinetics of melphalan hydrochloride via injection
using a propylene glycol diluent compared with the pharmacokinetics
of melphalan hydrochloride via injection using a propylene
glycol-free diluent (i.e., a cyclodextrin derivative) in multiple
myeloma patients undergoing autologous stem cell
transplantation.
[0233] Patients were screened prior to enrollment in the study.
Patients from any of the following classes were qualified for
inclusion in the study:
[0234] Patients with symptomatic multiple myeloma requiring
treatment at diagnosis or anytime thereafter;
[0235] Patients with multiple myeloma who qualify for autologous
stem cell transplantation therapy who have received appropriate
primary induction therapy for transplantation;
[0236] Patients who are 70 years of age or younger at time of
transplantation (patients greater than 70 years of age may qualify
on a case-by-case basis if the patient meets criterion based on
institution's standard of practice);
[0237] Patients with an adequate autologous graft, defined as an
un-manipulated, cryopreserved, peripheral, blood stern cell or bone
marrow stem cell graft containing at least 2.times.10.sup.6 CD34+
cells/kg based upon patient weight, along with a reserve of
2.times.10.sup.6 CD34+ cells/kg that is stored in a separate bag;
and
[0238] Patients with adequate organ function as Measured by: [0239]
Cardiac: Left ventricular ejection fraction at rest >40%; [0240]
Hepatic: Bilirubin <2.times. the upper limit of normal, and
ALT/AST <3.times.ULN; [0241] Renal: -Creatinine clearance >40
mL/minute; and [0242] Pulmonary: DLCQ, FEV.sub.1, PVC. >50% of
predicted value (corrected for Hgb) or O.sub.2 saturation >92%
on room air.
[0243] All patients have received antiemetics hydration, and
infection prophylaxis according to -institutional guidelines.
Patients, followed institutional guidelines regarding
hospitalization. Patients returned for daily laboratory tests (CBC
with differential and platelets and a basic chemistry panel) until
neutrophil engraftment, and then returned for weekly safety
evaluations until autologous stem cell transplantation Day +30. The
following safety, efficacy, and pharmacokinetic evaluations were
performed prior to the first: dose of melphalan, and at the
following post-dose time points: [0244] Twelve blood samples were
taken at specific time points for a pharmacokinetic evaluation.
Blood samples were collected immediately prior to and after
receiving the melphalan dose; [0245] Vital signs were recorded
hourly' during the first: eight hours after receiving each dose of
melphalan, then repeated once: daily until hospital. discharge, and
then weekly until Day +30. Weight were collected at hospital
discharge and at Day +30; [0246] A 12-lead electrocardiograph,
(ECG) along with a 10 to 20 second rhythm strip was collected twice
weekly until hospital discharge, then a 12-lead ECG (without a
rhythm strip), was collected weekly until Day +30; [0247] A focused
physical examination was performed daily until hospital discharge,
them a complete physical examination was performed weekly until Day
+30; [0248] Toxicity grading and evaluation for AEs/SAEs was
according to NC1-CTC AE Version 3.0 during the entire Study Period;
[0249] Complete blood count with differential and platelet count
was performed daily until neutrophil and platelet engraftment,
-then weekly until Day +30; [0250] Eastern Cooperative Group
performance status was examined at the time of hospital discharge,
then weekly until Day +30; [0251] Basic serum chemistry panel
(sodium, potassium, chloride; glucose; creatinine, bicarbonate, and
BUN) daily until neutrophil engraftment; [0252] Full serum
chemistry panel (sodium, potassium, chloride, magnesium,
bicarbonate, glucose, total protein, albumin, calciiim1, phosphate,
uric acid, BUN, creatinine, CPK, total bilirubin, alkaline
phosphate, LDH, SGOT, and SOFT) will be monitored weekly until Day
+30; [0253] Urinalysis (specific, gravity, pH, protein, glucose,
ketones, nitrite, IU3Cs, and WBCs) was monitored twice weekly until
hospital discharge, then weekly until Day .+-.30; and [0254]
Concomitant medications was recorded during the entire study
period.
[0255] A melphalan dose of 200 g/m.sup.2 was divided into two
separate, consecutive doses of 100 mg/m.sup.2 administered on two
separate days (Day -3 and Day -2) prior to the patients receiving
an autologous stem cell transplantation. For the calculation of
body surface area, actual body weight was used for patients who
weighed less than or between 100% to 130% of their ideal body
weight. Patients who weighed more than 130% of their ideal body
weight were dosed based on a body surface area obtained by
calculating the patient's adjusted body weight.
[0256] Patients were randomly chosen to receive, the first
melphalan Idose of 100 mg/m.sup.2 (on Day -3) via either a
composition comprising a propylene glycol diluent (i.e., Melphalan
IIC1 injectable, Bioniche Pharma USA) or a composition comprising,
a cyclodextrin derivative (SBE.sub.6.5-.beta.-CD, CAPTISOL.RTM., at
a concentration of 125 mM). Patients who randomly received the
first melphalan dose of 100 mg/m.sup.2 as a composition comprising
a cyclodextrin derivative (on Day -3) then received a second
melphalan dose of 100 mg/m.sup.2 (On Day -2) using the composition
comprising a: propylene glycol diluent. Conversely, patients: who
randomly received the first melphalan dose of 100 mg/m.sup.2 as a
composition comprising a propylene glycol diluent (on Day -3) then
received a second melphalan dose of 100 mg/m.sup.2 (on Day -2)
using the composition comprising a cyclodextrin derivative.
[0257] For the composition comprising a cyclodextrin derivative, a
dry powder composition containing melphalan as a hydrochloride salt
was diluted with normal saline to a melphalan concentration: no
greater than 0.45 mg/mL and a cyclodextrin concentration of 125 mM.
The diluted solution was infused over 60 minutes via a central
voicing catheter.
[0258] The composition comprising a propylene glycol diluent was
administered using a cyclodextrin-free composition (Melphalan HCl
Injectable, Bioniche Pharma USA) using the protocol described
herein supra.
[0259] Following one day of rest after the myeloablative
conditioning (Day -1). Patients received an autologous graft with a
minimum cell dose of 2.times.10.sup.6 CD34.sup.+ cells/kg of
patient; body weight (Day 0). Cryopreservation and thawing of: the
product was consistent with. Foundation for the Accreditation of
Cellular Therapy standards and local institutional practice. The
graft was infused per institutional, protocol. Starting on. Day +5,
G-CSF was be administered at a dose of 5 ug/kg/day until absolute
neutrophil count Was greater than 500/mm.sup.3:
[0260] Blood Samples for pharmacokinetic evaluation a melphalan
were collected after each dose of melphalan and the pharmacokinetic
parameters for in vivo melphalan distribution were evaluated.
Samples for evaluation of the pharmacokinetic parameters were
collected by taking 5 MT-venous blood samples immediately prior to
melphalan administration and at 0, 10, 20, 30, 60, 90, 120; 180,
240, 360; and 480 minutes following the end of the melphalan
infusion. Pharmacokinetic parameters were determined by
nonparametric pharmacokinetic data analysis: techniques:
Pharmacokinetic parameters computed from plasma drug:
concentration.-time data include the following; [0261] C.sub.max,
derived from the individual raw data; [0262] T.sub.max derived from
the individual raw data; [0263] Apparent terminal first-order
elimination rate constant (k.sub.el); [0264] Apparent elimination
t.sub.1/25; [0265] Area under the plasma concentration-time curve
to the last measurable time point (AUC.sub.0-1), calculated by the
trapezoidal rule; and [0266] Area under the plasma
concentration-time curve from the last measurable time point
extrapolated to infinity (AUC.sub.t-.infin.), determined from the
concentration at the last measurable time point divided by the
k.sub.el.
[0267] The plasma concentrations and pharmacokinetic 'parameters
were summarized using descriptive statistics, The data from
patients 1-3 is shown in the following table.
TABLE-US-00011 TABLE Individual Patient: melphalan pharmacokinetic:
parameters after intravenous administration of a. melphalan
formulation, that contained SBE.sub.6.5-.beta.-CD and a
cyclodextrin-free melphalan. formulation (i.e., Melphalan HCI
Injectable, Bioniche Phamta USA). SBF.45.57-13-CD CD-free Patient
Parameter (27% w/v) formulation Ratio 1.sup.a C.sub.max (ng/mL)
3,230 2,160 1.50 T.sub.max (min) 10 20 -- AUC.sub.0-t 259,073
202,714 1.28 AUC.sub.0-.infin. 264,656 208,028 1.27 .lamda..sub.z
(min.sup.-1) 0.0105 0.0103 -- T.sub.1/2 (min) 65.8 67.1 -- 2.sup.b
C.sub.max (ng/mL) 2,730 2,010 1.36 T.sub.max (min) 10 10 --
AUC.sub.0-t 198,051 151,456 1.31 AUC.sub.0-.infin. 202,728 154,130
1.32 .lamda..sub.z (min.sup.-1) 0.0103 0.0113 -- T.sub.1/2 (min)
67.4 61.6 -- 3.sup.a C.sub.max (ng/mL) 4,590 2890 1.59 T.sub.max
(min) 10 10 -- AUC.sub.0-t 306,432 230,681 1.33 AUC.sub.0-.infin.
314,108 336,059 1.33 .lamda..sub.z (min.sup.-1) 0.0104 0.0104 --
T.sub.1/2 (min) 68.5 66.4 -- .sup.aPatients 1 and 3 were
administered, the SBE.sub.6.5-.beta.-CD-containing formulation on
Day -3 and the CD-free formulation on Day. 72. .sup.bPatient 2 was
administered the CD-free formulation on Day -3 and the
SBE.sub.6.5-.beta.-CD - containing formulation on Day -2.
[0268] FIG. 6, provides a graphic representation of the wean plasma
melphalan concentration in a human patient after intravenous
administration of a melphalan formulation containing a cyclodextrin
derivative (SBE.sub.6.5-.beta.-CD) and after intravenous
administration of a cyclodextrin-free rnelphalan formulation:
(Melphalan HCl Injectable, Bioniche Pharrna USA). Referring to FIG.
6 and the data in the above table; the in vivo distribution of
melphalan administered with a sulfoalkyl ether cyclodextrin
derivative provides a nearly 50% increase the maximum in vivo
concentration of melphalan, and approximately a 30% increase, in
the area under the plasma conde-titration curves (i.e., for both
AUC.sub.0-t and AUC.sub.0-.infin.). As shown in. the table above,
the data for patients 2 and 3 exhibited similar pharmacokinetic
results. In view of pharmacokinetic data obtained for these
melphalan formulations in: the rat model the enhancement in
C.sub.max and AUC for the SBE.sub.6.5-.beta.-CD-containing
melphalan formulation in human patients is wholly unexpected.
[0269] As noted above, the study is on-going. The primary efficacy
end points, which will be based on an intent-to-treat analysis of
all patients; will be the rate of myeloablation and the rate
neutrophil engraftment. The following definitions will be used for
these endpoints:
[0270] Myeloablation will be defined as any: of the following:
[0271] Absolute neutrophil count less than 0.5.times.10.sup.9/L;
[0272] Absolute lymphocyte count less than. 0.1.times.10.sup.9/L or
[0273] Platelet count less than 20,000/mm.sup.3 or bleeding
requiring transfusion.
[0274] The first of two consecutive days for which cell counts drop
below these cut-off levels, will be recorded as the date of
myeloablation,
[0275] Neutrophil engraftment is defined as absolute neutrophil
count greater than 0.5.times.10.sup.9/L on three consecutive, daily
assessments.
[0276] Secondary efficacy end points will be based on the following
criteria: [0277] The rate of platelet engraftment, which will be
defined as an un-transfused platelet measurement
>20,000/mm.sup.3 on three consecutive daily assessments;| [0278]
The time to neutrophil engraftment, which will, be defined as the
first of three assessments where absolute neutrophil count is
greater than 0.5.times.10.sup.9/L;| [0279] The time to platelet
engraftment, which will be defined as the: first of three
consecutive daily assessments where un-transfused platelet
measurement is greater than 20,000/mm.sup.3;| [0280] The rate of
non-engraftment, which will be defined as a failure to reach an
absolute neutrophil count greater than: 0.5.times.10.sup.9/L. on
three consecutive: daily assessments by autologous stem cell
transplantation Day +100;| [0281] The rate of late graft failure or
late rejection, which will be defined as development of absolute
neutrophil: count less than 0.5.times.10.sup.9/L after having
engrafted within the first 100 days;| [0282] The rate of multiple
myeloma response (sCR, CR, VGPR, PR, SD, or PD); which, will be
defined according to International: Working Group: criteria at Day
+100; and| [0283] The rate of treatment-related mortality, which
will be defined as death without relapse or progression at Day
+100.
[0284] The clinical trial is expected to demonstrate that melphalan
administered with a cyclodextrin derivative (SBE.sub.6.5-.beta.-CD)
is therapeutically effective and safe for use in subjects for whom,
a stem cell transplantation has been indicated as conditioning
prior to stem cell transplantation.
Example 18
[0285] A Phase IIb, multi-center, open-label, non-randomized,
efficacy and safety study of melphalan hydrochloride administered
by injection using a propylene glycol-free vehicle will be
conducted in human multiple myeloma patients who have symptomatic
multiple myeloma and, qualify for ASCT.
[0286] The parameters of the study will be similar to those
described in' Example 17, except that all patients will be
administered a propylene glycol-free: melphalan composition (100
mg/m.sup.2) on Day -3 and Day -2 using a melphalan composition that
includes a cyclodextrin derivative. Otherwise, the inclusion
criteria, exclusion criteria, safety criteria, dosing, treatment,
and efficacy endpoints will be similar to those described above in
Example 17.
CONCLUSION
[0287] These examples illustrate possible embodiments of the
present invention. While various embodiments of the present
invention have been described above, it should be understood that
they have been presented by way of example only, and not limitation
it will be apparent to persons skilled in the relevant art that
various changes in farm and detail can be made therein without
departing from the spirit and scope: of the invention. Thus, the
breadth and scope of the present invention should not be limited by
any of the above-described, exemplary embodiments, but should be
defined only in accordance with the following claims, and their
equivalents.
[0288] All documents cited herein, including journal articles or
abstracts, published or: corresponding U.S. or foreign patent
applications, issued or foreign patent % or any other documents,
are each entirely incorporated by reference herein, including all
data, tables, figures, and text presented 'in the cited
documents.
* * * * *