U.S. patent application number 10/455999 was filed with the patent office on 2004-04-22 for injectable ready-to-use solutions containing an antitumor anthracycline glycoside.
This patent application is currently assigned to Pharmacia & Upjohn Company. Invention is credited to Bottoni, Giuseppe, Confalonieri, Carlo, De Ponti, Roberto, Gambini, Luciano, Gatti, Gaetano, Oldani, Diego.
Application Number | 20040077560 10/455999 |
Document ID | / |
Family ID | 27571230 |
Filed Date | 2004-04-22 |
United States Patent
Application |
20040077560 |
Kind Code |
A1 |
Gatti, Gaetano ; et
al. |
April 22, 2004 |
Injectable ready-to-use solutions containing an antitumor
anthracycline glycoside
Abstract
A sealed glass container containing therein a stable,
injectable, sterile, pyrogen-free doxorubicin anti-tumor
composition in a solution which consists essentially of a
physiologically acceptable salt of doxorubicin dissolved in a
physiologically acceptable solvent therefor, wherein said solution
has not been reconstituted from a lyophilizate, and wherein said
solution has a pH of from 2.5-3.5 and a concentration of said
doxorubicin of from 0.1 to 100 mg/ml.
Inventors: |
Gatti, Gaetano; (Sesto San
Giovanni (Milan), IT) ; Oldani, Diego; (Robecco sul
Naviglio ( Milan), IT) ; Bottoni, Giuseppe; (Bergamo,
IT) ; Confalonieri, Carlo; (Cusano Milanino (Milan),
IT) ; Gambini, Luciano; (Cornaredo (Milan), IT)
; De Ponti, Roberto; (Milan, IT) |
Correspondence
Address: |
Emily Miao
McDonnell Boehnen Hulbert & Berghoff
32nd Floor
300 S. Wacker Drive
Chicago
IL
60606
US
|
Assignee: |
Pharmacia & Upjohn
Company
|
Family ID: |
27571230 |
Appl. No.: |
10/455999 |
Filed: |
November 24, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10455999 |
Nov 24, 2003 |
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09902030 |
Jul 10, 2001 |
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6632799 |
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09902030 |
Jul 10, 2001 |
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09537638 |
Mar 29, 2000 |
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09537638 |
Mar 29, 2000 |
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09149381 |
Sep 8, 1998 |
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6087340 |
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09149381 |
Sep 8, 1998 |
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08368402 |
Jan 3, 1995 |
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5977082 |
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08368402 |
Jan 3, 1995 |
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08224993 |
Apr 8, 1994 |
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08224993 |
Apr 8, 1994 |
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07827938 |
Jan 29, 1992 |
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07827938 |
Jan 29, 1992 |
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07471005 |
Jan 25, 1990 |
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5124318 |
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07471005 |
Jan 25, 1990 |
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07341249 |
Apr 20, 1989 |
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07341249 |
Apr 20, 1989 |
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07064653 |
Jun 22, 1987 |
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07064653 |
Jun 22, 1987 |
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06878784 |
Jun 26, 1986 |
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Current U.S.
Class: |
514/34 |
Current CPC
Class: |
A61K 47/10 20130101;
A61K 47/12 20130101; C07H 15/252 20130101; A61K 9/0019 20130101;
A61K 47/26 20130101; A61K 47/02 20130101; A61K 47/18 20130101; A61K
31/704 20130101; A61K 31/70 20130101; A61K 9/08 20130101 |
Class at
Publication: |
514/034 |
International
Class: |
A61K 031/704 |
Foreign Application Data
Date |
Code |
Application Number |
Aug 2, 1985 |
GB |
8519452 |
Dec 5, 1986 |
GB |
8629193 |
Claims
What is claimed as new and desired to be secured by Letters of
patent of the United States is:
1-17. (Cancelled)
18. A physiologically acceptable solution of doxorubicin
hydrochloride dissolved in a physiologically acceptable solvent,
having a pH adjusted to from about 2.5 to 5.0 with a
physiologically acceptable acid and has a concentration of
doxorubicin of from 0.1 to 100 mg/ml.
19. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18, wherein said solution has not been
reconstituted from a lyophilizate.
20. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18, wherein said solution is contained in a
sealed container.
21. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18, wherein said solution has not been
reconstituted from a lyophilizate and is contained in a sealed
container.
22. A physiologically acceptable solution of doxorubicin
hydrochloride dissolved in a physiologically acceptable solvent,
having a pH adjusted to from 2.5 to 5.0 with a physiologically
acceptable acid selected from the group consisting of hydrochloric
acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric
acid, acetic acid, succinic acid, tartaric acid, ascorbic acid,
citric acid, glutamic acid, benzoic acid, methane sulfonic acid,
and ethane sulfonic acid and the concentration of said doxorubicin
hydrochloride being from 0.1 to 100 mg/ml.
23. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 22, wherein said solution has not been
reconstituted from a lyophilizate.
24. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 22, wherein said solution is contained in a
sealed container.
25. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 22, wherein said solution has not been
reconstituted from a lyophilizate and is contained in a sealed
container.
26. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein the physiologically
acceptable solvent is selected from the group consisting of water,
physiological saline, dextrose, polyethylene glycol,
N,N-dimethylacetamide, N-hydroxy-2-ethyl-lactamide, ethanol, benzyl
alcohol, propylene glycol, glycerin, diacetine, triacetine,
polyethylene glycol 400, propylene glycol methylether,
isopropylidenglycerin, dimethylisosorbide, 2-pyrrolidone,
N-methyl-2-pyrrolidone, Brij.RTM., Cremophor.RTM., Myrj.RTM.,
Tween.RTM. and Pluronics.RTM. and mixtures thereof.
27. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein the physiologically
acceptable solvent is water.
28. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein said physiologically
acceptable solvent is a saline solution.
29. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein said physiologically
acceptable solvent is a dextrose solution.
30. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein said physiologically
acceptable solvent is sterile water.
31. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, further comprising a tonicity
adjusting agent.
32. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein the concentration of
doxorubicin hydrochloride is from 0.1 to 50 mg/ml.
33. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein the concentration of
doxorubicin hydrochloride is from 1 to 20 mg/ml.
34. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein the pH of said solution is
from about 2.5 to about 3.5.
35. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein the pH of said solution is
from about 2.7 to about 3.3.
36. The physiologically acceptable solution of doxorubicin
hydrochloride of claim 18 or 22, wherein said solution is storage
stable, intravenously injectable, sterile and pyrogen-free.
37. A physiologically acceptable solution of anthracycline
glycoside selected from the group consisting of doxorubicin
hydrochloride, epirubicin hydrochloride and idarubicin
hydrochloride dissolved in a physiologically acceptable solvent,
having a pH adjusted to from about 2.5 to 5.0 with a
physiologically acceptable acid and has a concentration of
idarubicin of from 0.1 to 100 mg/ml.
38. The physiologically acceptable solution of anthracycline
glycoside of claim 37, wherein said solution has not been
reconstituted from a lyophilizate.
39. The physiologically acceptable solution of anthracycline
glycoside of claim 37, wherein said solution is contained in a
sealed container.
40. The physiologically acceptable solution of anthracycline
glycoside of claim 37, wherein said solution has not been
reconstituted from a lyophilizate and is contained in a sealed
container.
41. A physiologically acceptable solution of anthracycline
glycoside selected from the group consisting of doxorubicin
hydrochloride, epirubicin hydrochloride and idarubicin
hydrochloride dissolved in a physiologically acceptable solvent,
having a pH adjusted to from 2.5 to 5.0 with a physiologically
acceptable acid selected from the group consisting of hydrochloric
acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric
acid, acetic acid, succinic acid, tartaric acid, ascorbic acid,
citric acid, glutamic acid, benzoic acid, methane sulfonic acid,
and ethane sulfonic acid and the concentration of said idarubicin
hydrochloride being from 0.1 to 100 mg/ml.
42. The physiologically acceptable solution of anthracycline
glycoside of claim 41, wherein said solution has not been
reconstituted from a lyophilizate.
43. The physiologically acceptable solution of anthracycline
glycoside of claim 41, wherein said solution is contained in a
sealed container.
44. The physiologically acceptable solution of anthracycline
glycoside of claim 41, wherein said solution has not been
reconstituted from a lyophilizate and is contained in a sealed
container.
45. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein the physiologically acceptable
solvent is selected from the group consisting of water,
physiological saline, dextrose, polyethylene glycol,
N,N-dimethylacetamide, N-hydroxy-2-ethyl-lactamide, ethanol, benzyl
alcohol, propylene glycol, glycerin, diacetine, triacetine,
polyethylene glycol 400, propylene glycol methylether,
isopropylidenglycerin, dimethylisosorbide, 2-pyrrolidone,
N-methyl-2-pyrrolidone, Brij.RTM., Cremophor.RTM., Myrj.RTM.,
Tween.RTM. and Pluronics.RTM. and mixtures thereof.
46. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein the physiologically acceptable
solvent is water.
47. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein said physiologically acceptable
solvent is a saline solution.
48. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein said physiologically acceptable
solvent is a dextrose solution.
49. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein said physiologically acceptable
solvent is sterile water.
50. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 further comprising a tonicity adjusting
agent.
51. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein the concentration of
anthracycline glycoside selected from the group consisting of
doxorubicin hydrochloride, epirubicin hydrochloride and idarubicin
hydrochloride is from 0.1 to 50 mg/ml.
52. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein the concentration of
anthracycline glycoside selected from the group consisting of
doxorubicin hydrochloride, epirubicin hydrochloride and idarubicin
hydrochloride is from 1 to 20 mg/ml.
53. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein the pH of said solution is from
about 2.5 to about 3.5.
54. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein the pH of said solution is from
about 2.7 to about 3.3.
55. The physiologically acceptable solution of anthracycline
glycoside of claim 37 or 41 wherein said solution is storage
stable, intravenously injectable, sterile and pyrogen-free.
Description
[0001] This is a continuation of U.S. Ser. No. 09/902,030, filed
Jul. 10, 2001 which is a continuation of U.S. Ser. No. 09/537,638,
filed Mar. 29, 2000, now abandoned, which is a continuation of U.S.
Ser. No. 09/149,381, filed Sep. 8, 1998, now U.S. Pat. No.
6,087,340, which is a continuation of U.S. Ser. No. 08/368,402,
filed Jan. 3, 1995, now U.S. Pat. No. 5,977,082, which is a
continuation of U.S. Ser. No. 08/224,993, filed Apr. 8, 1994, now
abandoned, which is a continuation of U.S. Ser. No. 07/827,938,
filed Jan. 29, 1992, now abandoned, which is a divisional of U.S.
Ser. No. 07/471,005, filed Jan. 25, 1990, now U.S. Pat. No.
5,124,318, which is a continuation of U.S. Ser. No. 07/341,249,
filed Apr. 20, 1989, now abandoned, which is a continuation of U.S.
Ser. No. 07/064,653, filed Jun. 22, 1987, now abandoned, which is a
continuation-in-part of U.S. Ser. No. 06/878,784, filed Jun. 26,
1986, now abandoned.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates to a stable, injectable,
ready-to-use solution of an antitumor anthracycline glycoside, e.g.
doxorubicin (Adriamycin.RTM.), to a process for preparing such a
solution, and providing the same in a sealed container, and to a
method for treating tumors by the use of the ready-to-use
solution.
[0004] 2. Description of the Related Art
[0005] The anthracycline glycoside compounds are a well known class
of compounds in the antineoplastic group of agents, of which
doxorubicin is a typical, and the most widely used, representative:
Doxorubicin. Anticancer Antibiotics, Federico Arcamone, 1981, Publ:
Academic Press, New York, N. Y.; Adriamycin Review, EROTC
International Symposium, Brussels, May, 1974, edited by M. Staquet,
Publ. Eur. Press Medikon, Ghent, Belg.; Results of Adriamycin
Therapy, Adriamycin Symposium at Frankfurt/Main 1974 edited by M.
Ghione, J. Fetzer and H. Maier, publ.: Springer, New York, N.Y.
[0006] In the past, solutions of anthracycline glycosides have been
prepared and the stability thereof has been studied. However,
results of these studies have been inconsistent, and no clear
parameters have emerged for maintenance of a stable anthracycline
glycoside, e.g., doxorubicin, solution. Bosanquet, in a recent
article entitled "Stability of solutions of antineoplastic agents
during preparation and storage for in vitro assays," (Cancer
Chemother. Pharmacol. 1986, 17, 1-10) reviews the field of
stability studies, with particular emphasis on doxorubicin
(Adriamycin.RTM.). He points out that "very little can be
categorically stated about the stability of adriamycin, and a very
carefully designed study is urgently required to resolve these
conflicting results."
[0007] At present, anthracycline glycoside antitumor drugs, in
particular, e.g., doxorubicin, are solely available in the form of
lyophilized preparations, which need to be reconstituted before
administration.
[0008] Both the manufacturing and the reconstitution of such
preparations expose the involved personnel (workers, pharmacists,
medical personnel, nurses) to risks of contamination which are
particularly serious due to the toxicity of the antitumor
substances.
[0009] Indeed, the Martindale Extra Pharmacopoeia 28th edition,
page 175 left column, reports on adverse effects of antineoplastic
drugs and recommends that "They must be handled with great care and
contact with skin and eyes avoided; they should not be inhaled.
Care must be taken to avoid extravasation since pain and tissue
damage may ensue".
[0010] Similarly, Scand. J. Work Environ Health vol. 10(2), pages
71-74 (1984), as well as articles in Chemistry Industry, Issue Jul.
4, 1983, page 488, and Drug-Topics-Medical-Economics-Co, Issue Feb.
7, 1983, page 99, report severe adverse effects observed in medical
personnel exposed to use of cytostatic agents, including
doxorubicin.
[0011] Even though the effect of long-term low-level exposure to
such cytotoxic drugs is not yet completely known, there is
certainly a hazard for those who regularly prepared and administer
these substances in view of the fact that they are known mutagens
and carcinogens in animals and implicated as carcinogens in
man.
[0012] To administer a lyophilized preparation, double handling of
the drug is required, the lyophilized cake having to be first
reconstituted and then administered. Moreover, in some cases, the
complete dissolution of the powder may require prolonged shaking
because of solubilization problems. Reconstitution of a lyophilized
cake or powder can result in formation of aerosol droplets which
can be inhaled or can come into contact with skin or mucous
membranes of those handling the solution.
SUMMARY OF THE INVENTION
[0013] As the risks connected with the manufacturing and the
reconstitution of a lyophilized preparation would be highly reduced
if a ready-to-use solution of the drug were available, the present
inventors have developed a stable, therapeutically acceptable
injectable solution of an anthracycline glycoside drug, e.g.
doxorubicin, whose preparation and administration does not require
either lyophilization or reconstitution.
[0014] According to the present invention, there is provided a
stable, injectable, sterile, pyrogen-free, anthracycline glycoside
solution which consists essentially of a physiologically acceptable
salt of an anthracycline glycoside dissolved in a physiologically
acceptable solvent therefor, which has not been reconstituted from
a lyophilizate, which has a pH of from 2.5 to 3.5 and which is
preferably contained in a sealed glass container.
DESCRIPTION OF THE FIGURES
[0015] FIG. 1 is a K.sub.obs pH profile for doxorubicin.HCl
degradation at 55.degree. C. in sterile water.
[0016] FIG. 2 is a K.sub.obs pH profile for doxorubicin.HCl
degradation at 55.degree. C. in 5% dextrose.
[0017] FIG. 3 is a K.sub.obs pH profile for doxorubicin.HCl
degradation at 55.degree. C. in 0.9% saline.
[0018] FIG. 4 is a logarithmic plot of residual doxorubicin.HCl
concentration versus time.
[0019] FIG. 5 is a logarithmic plot of residual doxorubicin.HCl
concentration in 0.9% NaCl.
[0020] FIG. 6 is a logarithmic plot of residual doxorubicin.HCl
concentration in 5% dextrose.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0021] A preferred pH range for the anthracycline glycoside
solution of this invention is from 2.5 to 3.5. A more preferred
range is from 2.7 to 3.5, and a particularly preferred range is
from 2.7 to 3.3. A range of 2.7 up to about 3.0 may also be
mentioned as a useful range.
[0022] The preferred anthracycline glycoside is doxorubicin
hydrochloride. It is known that doxorubicin hydrochloride is more
stable in acidic than neutral or basic solutions. See Analytical
Profiles of Drug Substances, Vol. 9, Klaus Florey, ed. (Academic
Press 1980). A number of stability studies are summarized in
Bosanquet, Cancer Chemother. Pharmacol. 17, 1986, 1-10. However,
these studies are inconsistent, in part because of the varying
media used to make up the solutions and the methods used to measure
stability. Taken as a whole, the prior art has not appreciated with
any degree of certainty how to prepare a stable, injectable
doxorubicin solution, as a function of pH.
[0023] Martindale--The Extra Pharmacopeia 28th edition, 1828, on
page 205, indicates that a 0.5% solution of doxorubicin
hydrochloride and water has a pH of 3.8 to 6.5. Reconstitution of
the commercial freeze dried formulation which is made of
doxorubicin hydrochloride and lactose, leads to a solution having a
pH in the range of between 4.5 and 6, containing doxorubicin at 2
milligrams per milliliter concentration and lactose, and
additionally containing sodium chloride when saline is used for
reconstitution. In order to lower the pH below that of
reconstituted solutions, one must add an acid to lower the pH. In
the past there was little motivation for a user of the drug to add
an acid to lower the pH, since it was not recognized that the drug
was actually more stable at pH's between about 2.5 and 3.5.
According to the present invention, it has been discovered that
anthracycline glycosides, such as doxorubicin hydrochloride, are
stable in the pH range disclosed herein in physiologically
acceptable media. The prior art did not recognize stability for an
injectable doxorubicin solution in the particular narrow pH range
disclosed herein, so this pH range was not considered to be a
particularly useful range for administration of this drug.
[0024] Preferably the solution of the invention is provided in a
sealed container, especially one made of glass, either in a unit
dosage form or in a multiple dosage form.
[0025] In addition to doxorubicin, other anthracycline glycosides
include 4'-epi-doxorubicin (i.e. epirubicin), 4'-desoxy-doxorubicin
(i.e. esorubicin), 4'-desoxy-4'-iodo-doxorubicin, daunorubicin and
4-demethoxydaunorubicin (i.e. idarubicin).
[0026] Any physiologically acceptable salt of the anthracycline
glycoside may be used for preparing the solution of the invention.
Examples of suitable salts may be, for instance, the salts with
mineral inorganic acids such as hydrochloric, hydrobromic,
sulfuric, phosphoric, nitric and the like, and the salts with
certain organic acids such as acetic, succinic, tartaric, ascorbic,
citric, glutamic, benzoic, methanesulfonic, ethanesulfonic and the
like. The salt with hydrochloric acid is a particularly preferred
salt, especially when the anthracycline glycoside is
doxorubicin.
[0027] Any solvent which is physiologically acceptable and which is
able to dissolve the anthracycline glycoside salt may be used. The
solution of the invention may also contain one or more additional
components such as a co-solubilizing agent (which may be the same
as a solvent), a tonicity adjustment agent, a stabilizing agent and
a preservative. Examples of solvents, co-solubilizing agents,
tonicity adjustment agents, stabilizing agents and preservatives
which can be used for the preparation of the anthracycline
glycoside solutions of the invention are hereunder reported.
[0028] Suitable solvents and co-solubilizing agents may be, for
instance, water; physiological saline; aliphatic amides, e.g.
N,N-dimethylacetamide, N-hydroxy-2-ethyl-lactamide and the like;
alcohols, e.g. ethanol, benzyl alcohol and the like; glycols and
polyalcohols, e.g. propyleneglycol, glycerin and the like; esters
of polyalcohols, e.g. diacetine, triacetine and the like;
polyglycols and polyethers, e.g. polyethyleneglycol 400,
propyleneglycol methylethers and the like; dioxolanes, e.g.
isopropylidenglycerin and the like; dimethylisosorbide; pyrrolidone
derivatives, e.g. 2-pyrrolidone, N-methyl-2-pyrrolidone,
polyvinylpyrrolidone (co-solubilizing agent only) and the like;
polyoxyethylenated fatty alcohols, e.g., Brij.RTM. and the like;
esters of polyoxyethylenated fatty acids, e.g., Cremophor.RTM.,
Myrj.RTM. and the like; polysorbates, e.g., Tweens.RTM.,
polyoxyethylene derivatives of polypropyleneglycols, e.g.,
Pluronics.RTM..
[0029] A particularly preferred co-solubilizing agent is
polyvinylpyrrolidone.
[0030] Suitable tonicity adjustment agents may be, for instance,
physiologically acceptable inorganic chlorides, e.g. sodium
chloride; dextrose; lactose; mannitol; sorbitol and the like.
[0031] Preservatives suitable for physiological administration may
be, for instance, esters of parahydroxybenzoic acid (e.g., methyl,
ethyl, propyl and butyl esters, or mixtures of them), chlorocresol
and the like.
[0032] Suitable stabilizing agents include monosaccharides (e.g.,
galactose, fructose, and fucose), disaccharides (e.g., lactose),
polysaccharides (e.g., dextran), cyclic oligosaccharides (e.g.,
.alpha.-, .beta.-, .gamma.-cyclodextrin), aliphatic polyols (e.g.,
mannitol, sorbitol, and thioglycerol), cyclic polyols (e.g.
inositol) and organic solvents (e.g., ethyl alcohol and glycerol).
These may be included in concentrations of from about 0.25-10% w/v,
preferably 0.5-5% w/v in the solution.
[0033] The above mentioned solvents and co-solubilizing agents,
tonicity adjustment agents, stabilizing agents and preservatives
can be used alone or as a mixture of two or more of them.
[0034] Examples of preferred solvents are water, ethanol,
polyethylene glycol and dimethylacetamide as well as mixtures in
various proportions of these solvents. Water is a particularly
preferred solvent. Other solvents giving good results in terms of
stability are 0.9% sodium chloride solution (i.e., physiological
saline), and, especially, 5% dextrose solution, 5% mannitol
solution and 5% sorbitol solution, i.e., aqueous solutions
containing approximately 5% of, respectively, dextrose, mannitol or
sorbitol. Small variations (.+-.2-3%) of these additional
ingredients also fall within the scope of the present
invention.
[0035] To adjust the pH within the range of from 2.5 to about 3.5,
a physiologically acceptable acid is added to the solution of the
anthracycline glycoside. The acid should be a physiologically
acceptable acid, e.g., an inorganic mineral acid such as
hydrochloric, hydrobromic, sulfuric, phosphoric, nitric and the
like, or an organic acid such as acetic, succinic, tartaric,
ascorbic, citric, glutamic, benzoic, methanesulphonic,
ethanesulfonic and the like, or also an acidic physiologically
acceptable buffer solution, e.g., a chloride buffer, an acetate
buffer, a phosphate buffer and the like.
[0036] In the solutions of the invention the concentration of the
anthracycline glycoside may vary within broad ranges, preferably
from 0.1 mg/ml to 100 mg/ml, in particular from 0.1 mg/ml to 50
mg/ml, most preferably from 1 mg/ml to 20 mg/ml.
[0037] The preferred ranges of concentration may be slightly
different for different anthracycline glycosides. Thus, for
example, preferred concentrations for doxorubicin are from about 2
mg/ml to about 50 mg/ml, preferably from 2 mg/ml to 20 mg/ml,
particularly appropriate values being 2 mg/ml and 5 mg/ml. Similar
concentrations are preferred also for 4'-epi-doxorubicin,
4'-desoxy-doxorubicin and 4'-desoxy-4'-iodo-doxorubic- in.
Preferred ranges of concentration for daunorubicin and
4-demethoxy-daunorubicin are from 0.1 mg/ml to 50 mg/ml, preferably
from 1 mg/ml to 20 mg/ml, concentrations of 1 mg/ml and 5 mg/ml
being particularly appropriate.
[0038] Suitable packaging for the anthracycline glycoside solutions
may be all approved containers intended for parenteral use, such as
plastic and glass containers, ready-to-use syringes and the like.
Preferably the container is a sealed glass container, e.g. a vial
or an ampoule. A hermetically sealed glass vial is particularly
preferred.
[0039] According to a particularly preferred feature of the
invention, there is provided, in a sealed glass container, a
sterile, pyrogen-free, injectable doxorubicin solution which
consists essentially of a physiologically acceptable salt of
doxorubicin dissolved in a physiologically acceptable solvent
therefor, which has not been reconstituted from a lyophilizate and
which has a pH of from 2.5 to 3.5.
[0040] In the above indicated preferred feature of the invention
the physiologically acceptable salt of doxorubicin may be, e.g. the
salt with a mineral inorganic acid such as hydrochloric,
hydrobromic, sulfuric, phosphoric, nitric and the like, or the salt
with an organic acid such as acetic, succinic, tartaric, ascorbic,
citric, glutamic, benzoic, methanesulfonic, ethanesulfonic and the
like. The hydrochloride salt is a particularly preferred salt.
[0041] For the solution hereabove indicated as a preferred feature
of the invention suitable solvents, co-solubilizing agents,
tonicity adjustment agents, stabilizing agents and preservatives
may be the same as those previously recited in this specification.
Water is a particularly preferred solvent.
[0042] Also, the physiologically acceptable acid which is added to
adjust the pH to from 2.5 to about 3.5 may be one of those
previously specified. When it is desired to adjust the pH of the
above said preferred solution to a value of from 2.5 to about 3.5,
hydrochloric acid is an especially preferred acid. Preferred pH
values for the above said preferred solutions of the invention are
from about 2.7 to about 3.3.
[0043] Though the concentration of doxorubicin in the above
preferred feature may vary within the broad range from 0.1 mg/ml to
100 mg/ml, preferred concentrations are from 2 mg/ml to 50 mg/ml,
most preferably from 2 mg/ml to 20 mg/ml: examples of especially
preferred concentrations of doxorubicin are 2 mg/ml and 5
mg/ml.
[0044] The invention also provides a process for producing a
sterile, pyrogen-free anthracycline glycoside solution with a pH of
from 2.5 to 3.5 which process comprises dissolving a
physiologically acceptable salt of the anthracycline glycoside,
which salt is not in the form of a lyophilizate, in a
physiologically acceptable solvent therefor; adding a
physiologically acceptable acid or buffer to adjust the pH within
the said range as desired; and passing the resulting solution
through a sterilizing filter.
[0045] One or more additional components such as co-solubilizing
agents, tonicity adjustment agents, stabilizing agents and
preservatives, for instance of the kind previously specified, may
be added to the solution prior to passing the solution through the
sterilizing filter.
[0046] With the solutions of the invention it is possible to obtain
compositions having a very high concentration of the anthracycline
glycoside active substance even at 50 mg/ml and more. This
constitutes a great advantage over the presently available
lyophilized preparations wherein high concentrations of
anthracycline glycoside can only be obtained with difficulty
because of solubilization problems encountered in reconstitution,
mainly with saline. The presence of the excipient, e.g. lactose, in
the lyophilized cake, and its generally high proportion in respect
of the active substance, even up to 5 parts of excipient per part
of active substance, has a negative effect on solubilization so
that difficulties may arise in obtaining dissolution of the
lyophilized cake, especially for concentrations of anthracycline
glycoside higher than 2 mg/ml.
[0047] The solution of the invention are characterized by a good
stability. Solutions in various solvents and with different pH's
and concentrations have been found to be stable for long periods at
temperatures accepted for the storage of pharmaceutical
preparations. This is illustrated in the Examples which follow.
[0048] Due to the well known anti-tumor activity of the
anthracycline glycoside active drug substance, the pharmaceutical
compositions of the invention are useful for treating tumors in
both human and animal hosts. Examples of tumors that can be treated
are, for instance, sarcomas, including osteogenic and soft tissue
sarcomas, carcinomas, e.g., breast-, lung-, bladder-, thyroid-,
prostate- and ovarian carcinoma, lymphomas, including Hodgkin and
non-Hodgkin lymphomas, neuroblastoma, melanoma, myeloma, Wilms
tumor, and leukemias, including acute lymphoblastic leukemia and
acute myeloblastic leukemia. See Kirk-Othmer's Encyclopedia of
Chemical Technology, Volume 5, pages 478-479 (1979).
[0049] Examples of specific tumors that can be treated are Moloney
Sarcoma Virus, Sarcoma 180 Ascites, solid Sarcoma 180, gross
transplantable leukemia, L 1210 leukemia and lymphocytic P 388
leukemia.
[0050] Thus, according to the invention there is also provided a
method of inhibiting the growth of a tumor, in particular one of
those indicated above, which comprises administering to a host
suffering from said tumor an injectable solution according to the
invention containing the active drug substance in an amount
sufficient to inhibit the growth of said tumor.
[0051] The injectable solutions of the invention are administered
by rapid intravenous injection or infusion according to a variety
of possible dose schedules. A suitable dose schedule for
doxorubicin may be, for example, of 60 to 75 mg of active drug
substance per m.sup.2 of body surface given as a single rapid
infusion and repeated at 21 days; an alternative schedule may be of
30 mg/m.sup.2 per day by intravenous route for 3 days, every 28
days. Suitable dosages for 4'-epi-doxorubicin and
4'-desoxy-doxorubicin may be, for instance, of 75 to 90 mg/m.sup.2
given in a single infusion to be repeated at 21 days, and similar
dosages may be useful also for 4'-desoxy-4'-iodo-doxorubicin.
[0052] Idarubicin, i.e. 4-demethoxy-daunorubicin, may be
administered intravenously at a single dose of 13-15 mg/m.sup.2
every 21 days in the treatment of solid tumors, while in the
treatment of leukemias a preferred dose schedule is, e.g., of 10-12
mg/m.sup.2 day be intravenous route for 3 days, to be repeated
every 15-21 days; similar dosages may be followed also for
daunorubicin.
EXAMPLES
[0053] The invention now being generally described, the same will
be better understood by reference to certain specific examples
which are included herein for purposes of illustration only and are
not intended to be limiting of the invention or any embodiment
thereof, unless specified.
[0054] With reference to the first three examples, the stability
controls on the ready-to-use solutions were carried out by means of
high performance liquid chromatography (HPLC), at the following
experimental conditions:
[0055] Liquid chromatograph: Varian model 5010
[0056] Spectrophotometric detector: Knauer model 8700
[0057] Integrating recorder: Varian model CDS 401
[0058] Injection valve: Rheodyne model 7125 fitted with a 10 mcl
sample loop
[0059] Chromatographic column: Waters .mu.-Bondapak C18 (length=300
mm; inner diameter=3.9 mm; average particle size=10 mcm)
[0060] Column temperature: ambient (about 22.degree.
C..+-.2.degree. C.)
[0061] Mobile phase: water:acetonitrile (69:31 v/v) adjusted to pH
2 with phosphoric acid, filtered (sintered glass filter, 1 mcm or
finer porosity) and deaerated
[0062] Mobile phase flow rate: 1.5 ml/min
[0063] Analytical wavelength: 254.+-.1 nm
[0064] Integrating recorder sensitivity: 512
[0065] Chart speed: 1 cm/min
[0066] At these conditions, the peak of the anthracycline glycoside
showed a retention time of about 6 minutes.
[0067] The obtained results are reported in Tables accompanying
Examples 1-3.
[0068] The extrapolation of the analytical data in order to
determine the time when the 90% of the initial assay could be
expected (t.sub.90 value) was made following an Arrhenius plot.
[0069] This procedure of analytical data treatment is well known
and widely used and described in the art: see e.g., Chemical
Stability of Pharmaceuticals, Kennet A. Connors, Gordon L. Amidon,
Lloyd Kennon, Publ. John Wiley and Sons, New York, N.Y., 1979.
[0070] The term "teflon" recurring in the examples refers to
"Teflon.TM.".
Example 1
[0071]
1 Composition for 80 vials (for 1 vial) Doxorubicin.multidot.HCl
0.8 g (10 mg) Water for injections 0.4 l (5 ml) q.s. to
[0072] Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the
amount of water for injections, de-aerated by nitrogen bubbling.
The pH of the solution was not adjusted. Further de-aerated water
for injections was then added to bring the solution to its final
volume (0.40 l).
[0073] The solution was filtered through a 0.22 .mu.m microporous
membrane under nitrogen pressure. Volumes of 5 ml of the solution
were distributed into type I-colorless glass vials having 5/7 ml
capacity. The vials were then closed with chlorobutyl teflon-faced
rubber stoppers and sealed with aluminum caps.
[0074] The stability of the solutions in the vials was tested. The
vials were stored at temperatures of 55.degree. C., 45.degree. C.
and 35.degree. C. (accelerated stability controls) and at 4.degree.
C. for up to 3 weeks (55.degree. C.), 4 weeks (45.degree. C. and
35.degree. C.) and 12 weeks (4.degree. C.).
[0075] The stability data obtained, using high performance liquid
chromatography (HPLC) for the determination of potency, are
reported in the following Table 1:
2 TABLE 1 INITIAL VALUES Concentration: 1.994 mg/ml pH = 5.2
Relative % Assay: 100.0 TEMPERATURE 4.degree. C. 35.degree. C.
45.degree. C. 55.degree. C. TIME Conc. Rel. % Conc. Rel. % Conc.
Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assay mg/ml Assay
mg/ml Assay 1 1.992 99.9 1.917 96.1 1.768 88.7 1.493 75.0 2 1.843
92.4 1.618 81.1 1.166 58.5 3 1.774 89.0 1.506 75.5 0.830 41.6 4
1.974 99.0 1.720 86.3 1.393 69.9 12 1.980 99.3 t.sub.90 (days)
extrapolated according to Arrhenius equation: t.sub.90 at 4.degree.
C. = 815 days t.sub.90 at 8.degree. C. = 480 days
[0076] Similar stability data can be observed also for analogous
solutions containing either doxorubicin hydrochloride at 5 mg/ml
concentration, or 4'-epi-doxorubicin, 4'-desoxy-doxorubicin,
4'-desoxy-4'-iodo-doxorubicin, daunorubicin or
4-demethoxy-daunorubicin, as hydrochloride salts, at both 2 mg/ml
and 5 mg/ml concentration.
Example 2
[0077]
3 Composition for 80 vials (for 1 vial) Doxorubicin.multidot.HCl
0.8 g (10 mg) Hydrochloric acid 0.1 N pH = 3 (pH = 3) q.s. to Water
for injections 0.4 l (5 ml) q.s. to
[0078] Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the
amount of water for injections, de-aerated by nitrogen bubbling.
The hydrochloric acid was then added dropwise to adjust the pH of
the solution to 3. Further de-aerated water for injections was then
added to bring the solution to its final volume (0.4 l).
[0079] The solution was filtered through a 0.22 .mu.m microporous
membrane under nitrogen pressure. Volumes of 5 ml of the solution
were distributed into type I-colorless glass vials having 5/7 ml
capacity. The vials were then closed with chlorobutyl teflon-faced
rubber stoppers and sealed with aluminum caps.
[0080] The stability of the solutions in the vials was tested. The
vials were stored at temperatures of 55.degree. C., 45.degree. C.
and 35.degree. C. (accelerated stability controls) and at 4.degree.
C. for up to 3 weeks (55.degree. C.), 4 weeks (45.degree. C. and
35.degree. C.) and 12 weeks (4.degree. C.).
[0081] The stability data obtained, using high performance liquid
chromatography (HPLC) for the determination of potency, are
reported in the following Table 2:
4 TABLE 2 INITIAL VALUES Concentration: 1.992 mg/ml pH = 3.0
Relative % Assay: 100.0 TEMPERATURE 4.degree. C. 35.degree. C.
45.degree. C. 55.degree. C. TIME Conc. Rel. % Conc. Rel. % Conc.
Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assay mg/ml Assay
mg/ml Assay 1 1.995 100.2 1.952 98.0 1.919 96.3 1.493 75.0 2 1.889
94.8 1.851 92.9 1.036 51.9 3 1.876 94.2 1.565 78.6 0.730 36.7 4
1.979 99.4 1.808 90.8 1.393 69.9 12 1.972 99.0 t.sub.90 (days)
extrapolated according to Arrhenius equation: t.sub.90 at 4.degree.
C. = 3970 days t.sub.90 at 8.degree. C. = 2000 days
[0082] Similar stability data can be observed also for analogous
solutions containing either doxorubicin hydrochloride at 5 mg/ml
concentration, or 4'-epi-doxorubicin, 4'-desoxy-doxorubicin,
4'-desoxy-4'-iodo-doxorubicin, daunorubicin or
4-demethoxy-daunorubicin, as hydrochloride salts, at both 2 mg/ml
and 5 mg/ml concentration.
Example 3
[0083]
5 Composition for 80 vials (for 1 vial) Doxorubicin.multidot.HCl
0.8 g (10 mg) Hydrochloric acid 0.1 N pH = 3 (pH = 3) q.s. to Water
for injections 0.4 l (5 ml) q.s. to
[0084] Doxorubicin.HCl (8.0 g) was dissolved in 90 percent of the
amount of water for injections, de-aerated by nitrogen bubbling.
The hydrochloric acid was then added dropwise to adjust the pH of
the solution 3. Further de-aerated water for injections was then
added to bring the solution to its final volume (0.4 l).
[0085] The solution was filtered through a 0.22 .mu.m microporous
membrane under nitrogen pressure. Volumes of 5 ml of the solution
were distributed into type I-colorless glass vials having 5/7 ml
capacity. The vials were then closed with chlorobutyl teflon-faced
rubber stoppers and sealed with aluminum caps.
[0086] The stability of the solutions in the vials was tested. The
vials were stored at temperatures of 55.degree. C., 45.degree. C.
and 35.degree. C. (accelerated stability controls) and at 4.degree.
C. for up to 3 weeks (55.degree. C.), 4 weeks (45.degree. C. and
35.degree. C.) and 12 weeks (4.degree. C.).
[0087] The stability data obtained, using high performance liquid
chromatography (HPLC) for the determination of potency, are
reported in the following Table 3:
6 TABLE 3 INITIAL VALUES Concentration: 20.06 mg/ml pH = 2.95
Relative % Assay: 100.0 TEMPERATURE 4.degree. C. 35.degree. C.
45.degree. C. 55.degree. C. TIME Conc. Rel. % Conc. Rel. % Conc.
Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assay mg/ml Assay
mg/ml Assay 1 20.06 100.0 19.56 97.5 17.84 88.9 12.31 61.4 2 18.87
94.1 15.61 77.8 7.09 35.3 3 18.24 90.9 13.41 66.8 3.13 15.6 4 19.91
99.2 17.51 87.3 11.07 55.2 12 19.80 98.7 t.sub.90 (days)
extrapolated according to Arrhenius equation: t.sub.90 at 4.degree.
C. = 3700 days t.sub.90 at 8.degree. C. = 1780 days
[0088] Similar stability data can be observed for analogous
solutions containing 4'-epi-doxorubicin or 4'-desoxy-doxorubicin,
as hydrochloride salts, at the same 20 mg/ml concentration.
[0089] The following examples regarding stability profile and
shelf-life forecasts were carried out under accelerated temperature
conditions on 5.0 ml of 2 mg/ml doxorubicin.HCl solutions in a
container-closure system consisting of: glass type I, 8 ml top
capacity vial; teflon-faced chlorobutyl rubber bung; aluminum
seal.
[0090] pH-Stability Profile at 55.degree. C. of Doxorubicin.HCl
Solutions in Sterile Water 5% Dextrose, 0.9% Saline
[0091] 2 mg/ml doxorubicin.HCl solutions were prepared in the
following 1=0.05 buffers: a) glycine.HCl pH 2.0, 2.5 and 3.0; b)
formate pH 3.5; c) acetate pH 4.0, 5.0 and 5.5.
[0092] 5.0 ml of each solution in glass vials were stored at
55.degree. C. and analyzed at prefixed times (up to 120 hours) for
doxorubicin.HCl assay and pH.
[0093] Tables 4, 5 and 6 give the doxorubicin.HCl residual
concentration and percent stability at 55.degree. C., at different
pH's and times of storage for sterile water, 5% dextrose and 0.9%
saline solutions, respectively.
[0094] The doxorubicin.HCl assays are the mean of three independent
determinations performed by the USP HPLC method (USP XXI). At each
pH value, the pseudo-first order rate constants (K.sub.obs) for the
degradation were calculated by linear regression analysis of the
natural logarithm of the residual concentration of doxorubicin.HCl
(.vertline.Dx.vertline..sub.t) versus time as depicted by the
following equation:
ln .vertline.Dx.vertline..sub.t=ln
.vertline.Dx.vertline..sub.o-K.sub.obs .sup..multidot.t
[0095] J. Thuro Carstensen, Theory of Pharmaceutical Systems,
Volume 1/General Principles, page 172, Academic Press, New York and
London 1972
[0096] Kenneth A. Connors, Gordon L. Amidon, Lloyd Kennon, Chemical
Stability of Pharmaceuticals, chapter 2, John Wiley and Sons, New
York 1979
[0097] Arthur Osol, Remington's Pharmaceutical Sciences, 16th
Edition, chapter 18, Mack Publishing Company, Easton, Pa. 1980
[0098] Valentino J. Stella, Chemical and Physical Bases Determining
the Instability and Incompatibility of Formulated Injectable Drugs,
Journal of Parenteral Sciences & Technology, July-August 1986,
page 142.
[0099] Tables 7, 8 and 9 give the observed rate constants
(K.sub.obs) for the degradation kinetics of doxorubicin.HCl at
55.degree. C. and at different pH's for sterile water, 5% dextrose
and 0.9% saline solutions, respectively.
[0100] FIGS. 1, 2, and 3 show the K.sub.obs-pH profile for the
doxorubicin.HCl degradation at 55.degree. C. in the above mentioned
media. The data in Tables 4-9 and the FIGS. 1-3 evidence that the 2
mg/ml doxorubicin.HCl solutions show at 55.degree. C. the maximum
stability in the pH range about 3.0-3.5 (.+-.0.2, e.g., 2.8, 3.2
and 3.3, 3.7) for all the three media tested. The range of from 2.5
to 3.0 is also a range of notable stability.
[0101] A common behavior as to stability is thus evidenced for
aqueous solutions in general, since no practical differences in
stability are observed when going from sterile water as such to
sterile water containing a tonicity adjustment agent, either ionic,
such as, e.g., sodium chloride, or non-ionic, such as, e.g.,
dextrose.
7TABLE 4 Accelerated (55.degree. C.) stability data of 2 mg/ml
doxorubicin.multidot.HCl solutions in sterile water at various pHs
Time (hours) Buffers Tests 0 8 16 24 48 72 120 pH 2.0
Doxorubicin.multidot.HCl 2.022 1.892 1.669 1.554 1.145 0.801
glycine-HCl assay .multidot. mg/ml .multidot. % stability 100.0
93.6 82.6 76.9 56.6 39.6 pH 2.00 2.01 2.02 2.01 2.01 2.02 pH 2.5
Doxorubicin.multidot.HCl assay 1.992 1.926 1.835 1.718 1.557 1.00
glycine-HCl mg/ml % stability 100.0 96.7 92.1 86.2 78.2 50.2 pH
2.51 2.50 2.50 2.52 2.51 2.52 pH 3.0 Doxorubicin.multidot.HCl assay
2.003 1.958 1.881 1.831 1.696 1.525 1.258 glycine-HCl mg/ml %
stability 100.0 97.8 93.9 91.4 84.7 76.1 62.8 pH 3.00 3.03 3.02
3.02 3.01 3.02 3.00 pH 3.5 Doxorubicin.multidot.HCl assay 2.035
1.950 1.887 1.840 1.650 1.538 1.241 formate mg/ml % stability 100.0
95.8 92.7 90.4 81.1 75.6 61.0 pH 3.51 3.51 3.51 3.51 3.52 3.52 3.51
pH 4.0 Doxorubicin.multidot.HCl assay 2.032 1.788 1.681 1.561 1.167
acetate mg/ml % stability 100.0 88.0 82.7 76.8 57.4 pH 4.00 4.00
4.04 4.02 4.02 pH 5.0 Doxorubicin.multidot.HC- l assay 2.019 1.823
1.688 1.512 1.060 acetate mg/ml stability 100.0 90.3 83.6 74.9 52.5
pH 5.03 5.05 5.04 5.04 5.05 pH 5.5 Doxorubicin.multidot.HCl assay
2.047 1.808 1.427 1.228 0.903 acetate mg/ml % stability 100.0 88.3
69.7 60.0 44.1 pH 5.50 5.53 5.53 5.54 5.56
[0102]
8TABLE 5 Accelerated (55.degree. C.) stability data of 2 mg/ml
doxorubicin.multidot.HCl solutions in 5% dextrose at various pHs
Time (hours) Buffers Tests 0 8 16 24 34 48 72 96 120 pH 2.0
Doxorubicin.multidot.HCl assay 1.993 1.851 1.683 1.513 1.361 1.078
0.765 glycine-HCl mg/ml % stability 100.0 92.8 84.4 75.9 68.3 54.1
38.4 pH 2.14 2.13 2.14 2.15 2.18 2.21 2.16 pH 2.5
Doxorubicin.multidot.HCl assay 1.967 1.897 1.822 1.760 1.682 1.499
1.305 glycine-HCl mg/ml % stability 100.0 96.4 92.6 89.5 85.5 76.2
66.3 pH 2.56 2.56 2.56 2.58 2.60 2.56 2.61 pH 3.0
Doxorubicin.multidot.HCl assay 1.975 1.908 1.832 1.645 1.508 1.344
1.206 glycine-HCl mg/ml % stability 100.0 96.6 92.7 83.3 76.4 68.0
61.1 pH 3.04 3.05 3.05 3.06 3.00 3.13 3.10 pH 3.5
Doxorubicin.multidot.HCl assay 1.983 1.897 1.858 1.622 1.324 1.222
formate mg/ml % stability 100.0 95.7 93.7 81.8 66.8 61.6 pH 3.58
3.59 3.60 3.63 3.60 3.63 pH 4.0 Doxorubicin.multidot.HCl assay
2.003 1.913 7.716 1.665 1.487 1.312 1.081 acetate mg/ml % stability
100.0 95.5 85.6 83.1 74.2 65.5 53.9 pH 4.10 4.10 4.11 4.11 4.16
4.15 4.12 pH 5.0 Doxorubicin.multidot.HCl assay 2.012 1.906 1.673
1.608 1.416 1.163 acetate mg/ml % stability 100.0 94.7 83.2 79.9
70.4 57.8 pH 5.06 5.06 5.06 5.06 5.07 5.04 pH 5.5
Doxorubicin.multidot.HC- l assay 1.991 1.841 1.470 1.246 1.091
acetate mg/ml % stability 100.0 92.5 73.8 62.6 54.8 pH 5.56 5.54
5.48 5.50 5.46
[0103]
9TABLE 6 Accelerated (55.degree. C.) stability data of 2 mg/ml
doxorubicin.multidot.HCl solutions in 0.9% saline at various pHs
Time (hours) Buffers Tests 0 4 8 16 24 34 48 72 96 120 pH 2.0
Doxorubicin.multidot.HCl assay 1.998 1.857 1.580 1.397 1.231 0.931
0.701 glycine-HCl mg/ml % stability 100.0 92.9 79.1 69.9 61.6 46.6
35.1 pH 2.16 2.16 2.18 2.16 2.22 2.20 2.19 pH 2.5
Doxorubicin.multidot.HCl assay 1.946 1.875 1.670 1.602 1.368 1.132
glycine-HCl mg/ml % stability 100.0 96.3 85.8 82.3 70.3 58.1 pH
2.59 2.59 2.59 2.58 2.62 2.62 pH 3.0 Doxorubicin.multidot.HCl assay
1.994 1.818 1.771 1.571 1.375 1.205 1.003 glycine-HCl mg/ml %
stability 100.0 91.2 88.8 78.8 69.0 60.4 50.3 pH 3.06 3.07 3.07
3.08 3.13 3.14 3.12 pH 3.5 Doxorubicin.multidot.HCl assay 1.997
1.824 1.742 1.543 1.323 1.176 0.919 formate mg/ml % stability 100.0
91.4 87.2 77.3 66.2 58.9 46.0 pH 3.58 3.56 3.56 3.66 3.61 3.64 3.63
pH 4.0 Doxorubicin.multidot.HCl assay 1.972 1.885 1.828 1.653 1.594
acetate mg/ml % stability 100.0 95.6 92.7 83.8 80.8 pH 4.10 4.10
4.10 4.10 4.11 pH 5.0 Doxorubicin.multidot.HCl assay 1.979 1.732
1.469 1.442 1.278 acetate mg/ml % stability 100.0 87.5 74.2 72.8
64.6 pH 5.04 5.06 5.04 5.05 5.05 pH 5.5 Doxorubicin.multidot.HCl
assay 2.023 1.847 1.548 1.330 acetate mg/ml % stability 100.0 91.3
76.5 65.7 pH 5.58 5.56 5.55 5.53
[0104]
10TABLE 7 K.sub.obs values (1/days) for the degradation of
doxorubicin.multidot.HCl 2 mg/ml solutions in sterile water at
various pHs at 55.degree. C. Buffer pH K.sub.obs .times. 10.sup.3
95% confidence limits Glycine-HCl 2.0 309.5 .+-.12.6 (I = 0.05)
Glycine-HCl 2.5 138.3 .+-.0.6 (I = 0.05) Glycine-HCl 3.0 93.1
.+-.4.6 (I = 0.05) Formate 3.5 96.7 .+-.4.4 (I = 0.05) Acetate 4.0
269.8 .+-.18.7 (I = 0.05) Acetate 5.0 322.6 .+-.19.2 (I = 0.05)
Acetate 5.5 415.4 .+-.45.7 (I = 0.05)
[0105]
11TABLE 8 K.sub.obs values (1/days) for the degradation of
doxorubicin.multidot.HCl 2 mg/ml solutions in 5% dextrose at
various pHs at 55.degree. C. Buffer pH K.sub.obs .times. 10.sup.3
95% confidence limits Glycine-HCl 2.0 323.8 .+-.17.2 (I = 0.05)
Glycine-HCl 2.5 138.7 .+-.9.9 (I = 0.05) Glycine-HCl 3.0 100.5
.+-.5.9 (I = 0.05) Formate 3.5 132.0 .+-.20.7 (I = 0.05) Acetate
4.0 209.7 .+-.12.7 (I = 0.05) Acetate 5.0 273.1 .+-.27.7 (I = 0.05)
Acetate 5.5 453.7 .+-.59.2 (I = 0.05)
[0106]
12TABLE 9 K.sub.obs values (1/days) for the degradation of
doxorubicin.multidot.HCl 2 mg/ml solutions in 0.9% saline at
various pHs at 55.degree. C. Buffer pH K.sub.obs .times. 10.sup.3
95% confidence limits Glycine-HCl 2.0 362.4 .+-.19.4 (I = 0.05)
Glycine-HCl 2.5 276.5 .+-.30.2 (I = 0.05) Glycine-HCl 3.0 133.2
.+-.8.0 (I = 0.05) Formate 3.5 148.1 .+-.11.1 (I = 0.05) Acetate
4.0 215.7 .+-.35.4 (I = 0.05) Acetate 5.0 301.2 .+-.60.1 (I = 0.05)
Acetate 5.5 430.3 .+-.69.9 (I = 0.05)
[0107] Shelf-Life (t 90%) Forecast of Doxorubicin.HCl 2 mg/ml
Sterile Water Solution Adjusted to pH 3.0
[0108] 5.0 ml of doxorubicin.HCl 2 mg/ml aqueous solution adjusted
to pH 3.0 with 0.5 N HCl were stored, in glass vials, at:
[0109] a) 55.degree. C. for 21 days, b) 45.degree. C. and
35.degree. C. for 28 days, c) 27.degree. C. for 90 days.
[0110] b) At prefixed times the vials were analyzed for
doxorubicin.HCl assay and pH.
[0111] The logarithmic plots of the residual concentration versus
time were linear and indicated the degradation of the drug to
follow pseudo-first order kinetics at constant pH and
temperature.
[0112] The observed rate constants (K.sub.obs) for the degradation
were calculated again by linear regression analysis of a plot of
the natural logarithm of the residual concentration of
doxorubicin.HCl (.vertline.Dx.vertline..sub.t) versus time as
depicted by the equation previously reported:
ln .vertline.Dx.vertline..sub.t=ln
.vertline.Dx.vertline..sub.o-K.sub.obs .sup..multidot.t
[0113] The Arrhenius equation for the degradation process was
calculated from the K.sub.obs obtained from the different
temperatures taken in account for the testing (table 11).
[0114] Applying the equation, the rate constants for the
pseudo-first order reactions at 4.degree. C., 8.degree. C.,
15.degree. C. and 27.degree. C. were calculated, together with the
expected t.sub.90% at these temperatures.
[0115] Table 10 gives the doxorubicin.HCl residual concentration
and percent stability at pH 3.0, at different temperatures and
times of storage.
[0116] FIG. 4 gives the logarithm of the remaining doxorubicin.HCl
concentration versus time at different temperatures.
[0117] The t..sub.90% forecasts (table 11) show that a commercially
meaningful shelf-life can be attributed to doxorubicin.HCl 2 mg/ml
pH 3.0 aqueous solution only if the product is stored in a
refrigerator (between 2.degree. C. and 8.degree. C.).
13TABLE 10 Accelerated stability data of doxorubicin.multidot.HCl 2
mg/ml pH 3.0 solutions in sterile water at different times and
temperatures Storage Time (days) Temperature Tests 0 4 8 14 32 28
60 90 27.degree. C. Doxorubicin.multidot.HCl assay 1.992 1.993
1.988 1.962 1.941 1.908 1.850 mg/ml % stability 100.0 100.1 99.8
98.5 97.4 95.8 92.9 pH 3.00 2.95 2.94 2.95 2.94 2.96 2.93
35.degree. C. Doxorubicin.multidot.HCl assay 1.992 1.985 1.952
1.889 1.876 1.808 mg/ml % stability 100.0 99.6 98.0 94.8 94.2 90.8
pH 3.00 2.96 2.98 2.93 2.92 2.92 45.degree. C.
Doxorubicin.multidot.HCl assay 1.992 1.919 1.851 1.677 1.565 1.393
mg/ml % stability 100.0 96.3 92.9 84.2 78.6 69.9 pH 3.00 2.97 2.95
2.85 2.92 2.90 55.degree. C. Doxorubicin.multidot.HCl assay 1.992
1.760 1.493 1.036 0.730 mg/ml % stability 100.0 88.4 74.9 52.0 36.6
pH 3.00 2.94 2.90 2.80 2.82
[0118]
14TABLE 11 Arrhenius approach. Pseudo-first order rate constants,
Arrhenius equation, calculated t.sub.90% PSEUDO-FIRST ORDER RATE
CONSTANTS, OBSERVED VALUES (K.sub.obs) Temperature K.sub.obs
.times. 10.sup.3 (1/days) Correlation Coefficient 27.degree. C.
0.850 0.986 35.degree. C. 3.506 0.983 45.degree. C. 12.790 0.995
55.degree. C. 49.340 0.995
[0119] Arrhenius Equation from 27.degree. C., 35.degree. C.,
45.degree. C., and 55.degree. C. Rate Constants
[0120] In K.sub.obs=-14083/T+39.95
[0121] correlation coefficient=0.9988
[0122] Pseudo-First Order Rate Constants, Calculated Values (K)
15 Temperature K .times. .multidot. 10.sup.3 (1/days) t.sub.90%
(days) 95% confidence limits 4.degree. C. 0.019 5,652 3,079-10,380
8.degree. C. 0.038 2,745 1,603-4,697 15.degree. C. 0.130 810
532-1,238 27.degree. C. 0.918 115 89-147
[0123] Shelf-Life (t 90%) Forecast of Doxorubicin.HCl 2 mg/ml 0.9%
Sodium Chloride Solution Adjusted to pH 3.0
[0124] 5.0 ml of doxorubicin.HCl 2 mg/ml solution in 0.9% sodium
chloride adjusted to pH 3.0 with 0.5 N HCl were stored, in glass
vials, at: a) 60.degree. C. for 4 days, b) 55.degree. C. for 14
days, c) 45.degree. C. for 21 days, d) 35.degree. C. for 28
days.
[0125] At prefixed times the vials were analyzed for
doxorubicin.HCl assay and pH.
[0126] The logarithmic plots of the residual concentration versus
time were linear and indicated the degradation of the drug to
follow pseudo-first order kinetics at constant pH and
temperature.
[0127] The observed rate constants (K.sub.obs) for the degradation
were calculated again by linear regression analysis of a plot of
the natural logarithm of the residual concentration of
doxorubicin.HCl (.vertline.Dx.vertline..sub.t) versus time as
depicted by the equation previously reported:
ln .vertline.Dx.vertline..sub.t=ln
.vertline.Dx.vertline..sub.o-K.sub.obs .sup..multidot.t
[0128] The Arrhenius equation for the degradation process was
calculated from the K.sub.obs obtained from the different
temperatures taken in account for the testing (table 13).
[0129] Applying the equation, the rate constants for the
pseudo-first order reactions at 4.degree. C., 8.degree. C.,
15.degree. C. and 27.degree. C. were calculated, together with the
expected t..sub.90% at these temperatures.
[0130] Table 12 gives the doxorubicin.HCl residual concentration
and percent stability at pH 3.0, at different temperatures and
times of storage.
[0131] FIG. 5 gives the logarithm of the remaining doxorubicin.HCl
concentration versus time at different temperatures.
16TABLE 12 Accelerated stability data of doxorubicin.multidot.HCl 2
mg/ml in 0.9% sodium chloride at different times and temperatures
Storage Time (days) Conditions Tests 0 1 2 3 4 8 11 14 21 28
35.degree. C. Doxorubicin.multidot.HCl assay 2.061 2.045 1.946
1.932 1.852 mg/ml % stability 100.0 99.2 94.4 93.7 89.9 pH 3.05
2.98 2.92 2.92 2.98 45.degree. C. Doxorubicin.multidot.HCl assay
2.061 1.996 1.724 1.517 1.344 mg/ml % stability 100.0 96.5 83.6
73.6 65.2 pH 3.05 2.98 2.97 2.98 2.93 55.degree. C.
Doxorubicin.multidot.HCl assay 2.061 1.450 1.066 0.900 mg/ml %
stability 100.0 70.4 51.7 43.7 pH 3.05 2.90 2.97 2.95 60.degree. C.
Doxorubicin.multidot.HCl assay 2.061 1.742 1.481 1.290 1.050 mg/ml
% stability 100.0 84.5 71.9 62.6 50.9 pH 3.05 2.97 2.96 2.98
2.96
[0132] The t.sub.90% forecasts (table 13) show that a commercially
meaningful shelf-life can be attributed to doxorubicin.HCl 2 mg/ml
pH 3.0 solution in 0.9% sodium chloride only if the product is
stored in a refrigerator (between 2.degree. C. and 8.degree.
C.).
[0133] Table 13--Doxorubicin.HCl 2 mg/ml pH 3.0 Solution in 0.9%
NaCl
[0134] Arrhenius approach. Pseudo-first order rate constants,
[0135] Arrhenius equation, calculated t.sub.90%
17 PSEUDO-FIRST ORDER RATE CONSTANTS, OBSERVED VALUES (K.sub.obs)
Temperature K.sub.obs .times. 10.sup.3 (1/days) Correlation
Coefficient 35.degree. C. 3.89 0.965 45.degree. C. 21.61 0.987
55.degree. C. 75.90 0.996 60.degree. C. 164.90 0.998
[0136] Arrhenius Equation from 35.degree. C., 45.degree. C.,
55.degree. C. and 60.degree. C. Rate Constants
ln K.sub.obs=-15100/T+43.53
[0137] correlation coefficient=0.9986
18 PSEUDO-FIRST ORDER RATE CONSTANTS, CALCULATED VALUES (K)
Temperature K .times. 10.sup.3 (1/days) t.sub.90% (days) 95%
confidence limits 4.degree. C. 0.017 6,166 1,670-22,756 8.degree.
C. 0.037 2,838 861-9,351 15.degree. C. 0.137 768 281-2,105
27.degree. C. 1.112 94 45-197
[0138] Shelf-Life (t 90%) Forecast of Doxorubicin.HCl 2 mg/ml
Solution in 5% Dextrose Adjusted to pH 3.0
[0139] 5.0 ml of doxorubicin.HCl 2 mg/ml solution in 5% dextrose
adjusted to pH 3.0 with 0.5 N HCl were stored, in glass vials, at:
a) 60.degree. C. for 8 days, b) 55.degree. C. for 17 days, c)
45.degree. C. and 35.degree. C. for 28 days.
[0140] At prefixed times the vials were analyzed for
doxorubicin.HCl assay and pH.
[0141] The logarithmic plots of the residual concentration versus
time were linear and indicated the degradation of the drug to
follow pseudo-first order kinetics at constant pH and temperature.
The observed rate constants (K.sub.obs) for the degradation was
calculated again by linear regression analysis of a plot of the
natural logarithm of the residual concentration of doxorubicin.HCl
(.vertline.Dx.vertline..sub.t) versus time as depicted by the
equation previously reported:
ln .vertline.Dx.vertline..sub.t=ln
.vertline.Dx.vertline..sub.o.sup.-K.sub- .obs.sup..multidot.t
[0142] The Arrhenius equation for the degradation process was
calculated from the K.sub.obs obtained from the different
temperatures taken in account for the testing (table 15).
[0143] Applying the equation, the rate constants for the
pseudo-first order reactions at 4.degree. C., 8.degree. C., and
15.degree. C. and 27.degree. C. were calculated, together with the
expected t.sub.90% at these temperatures.
[0144] Table 14 gives the doxorubicin.HCl residual concentration
and percent stability at pH 3.0, at different temperatures and
times of storage.
[0145] FIG. 6 gives the logarithm of the remaining doxorubicin.HCl
concentration versus time at different temperatures.
[0146] The t.sub.90% forecasts (table 15) show that a commercially
meaningful shelf-life can be attributed to doxorubicin.HCl 2 mg/ml
pH 3.0 5% dextrose solution only if the product is stored in a
refrigerator (between 2.degree. C. and 8.degree. C.).
19TABLE 14 Accelerated stability data of doxorubicin.multidot.HCl 2
mg/ml pH 3.0 solution in 5% dextrose at different times and
temperatures Storage Time (days) Conditions Tests 0 2 4 6 8 11 14
17 21 28 35.degree. C. Doxorubicin.multidot.HCl assay 2.114 2.044
2.034 2.015 1.934 1.859 mg/ml % stability 100.0 96.7 96.2 53.3 91.5
87.9 pH 3.02 2.98 2.94 2.95 2.90 2.94 45.degree. C.
Doxorubicin.multidot.HCl assay 2.114 1.940 1.870 1.684 1.510 1.410
mg/ml % stability 100.0 91.8 88.5 79.7 71.5 66.7 pH 3.02 2.97 2.98
2.95 2.96 2.96 55.degree. C. Doxorubicin.multidot.HCl assay 2.114
1.718 1.415 1.112 0.957 0.796 mg/ml % stability 100.0 81.3 66.9
52.6 45.3 37.7 pH 3.02 2.95 2.92 2.99 2.91 2.95 60.degree. C.
Doxorubicin.multidot.HCl assay 2.114 1.752 1.393 1.176 0.925 mg/ml
% stability 100.0 82.9 65.9 55.7 43.8 pH 3.02 2.96 2.98 2.96
2.97
[0147] Table 15--Doxorubicin.HCl 2 mg/ml pH 3.0 Solution in 5%
Dextrose.
[0148] Arrhenius approach. Pseudo-first order rate constants,
[0149] Arrhenius equation, calculated t.sub.90%
20 PSEUDO-FIRST ORDER RATE CONSTANTS, OBSERVED VALUES (K.sub.obs)
Temperature K.sub.obs .times. 10.sup.3 (1/days) Correlation
Coefficient 35.degree. C. 4.190 0.990 45.degree. C. 14.55 0.995
55.degree. C. 58.11 0.998 60.degree. C. 102.6 0.999
[0150] Arrhenius Equation from 35.degree. C., 45.degree. C.,
55.degree. C. and 60.degree. C. Rate Constants
ln K.sub.obs=-13266/T+37.56
[0151] correlation coefficient=0.9993
21 PSEUDO-FIRST ORDER RATE CONSTANTS, CALCULATED VALUES (K)
Temperature K .times. 10.sup.3 (1/days) t.sub.90% (days) 95%
confidence limits 4.degree. C. 0.0326 3,218 1,463-7,082 8.degree.
C. 0.0645 1,628 792-3,344 15.degree. C. 0.203 516 281-949
27.degree. C. 1.283 82 53-128
[0152] Long Term Stability of Doxorubicin Formulations having a pH
Falling within the Range from 2.5 to 3.5
[0153] Batches tested, formulations tested and packaging used are
reported on, respectively, tables 16, 17 and 18, as well as on,
respectively, tables 25, 26 and 27.
[0154] Test and Methods
[0155] The formulations were tested as regards appearance, clarity
of solution, pH, sterility (8.degree. C., yearly), doxorubicin.HCl
assay.
[0156] Test Methods
[0157] For appearance and clarity; visual inspection
[0158] For pH: USP XXI
[0159] For sterility; USP XXI (membrane filtration)
[0160] For doxorubicin.HCl assay: HPLC ion-pair method and USP HPLC
method (USP XXI)
[0161] Brief description of the HPLC ion-pair method for
doxorubicin.
[0162] HCl assay:
[0163] Column filling: reverse phase, Zortax TMS
[0164] Mobile phase: water, acetonitrile, methanol (54:29:17 v/v/v)
containing 2 ml/l 85% phosphoric acid and 1 mg/ml sodium
laurylsulfate (pairing agent) adjusted to pH 3.5 with 2N NaOH
[0165] Mobile phase flow rate: 1.5 ml/min
[0166] Column temperature: ambient (22.degree. C..+-.2.degree.
C.)
[0167] Analytical wavelength: 254 nm
[0168] System suitability parameters: Symmetry factor between 0.7
and 1.2; number of theoretical plates .gtoreq.2500; measurement
reproducibility: variation coefficient <1, n=6; resolution
factor .gtoreq.12
[0169] The HPLC ion-pair method for doxorubicin.HCl assay is
validated for accuracy, precision, linearity, sensitivity,
specificity and stability-indicating nature.
[0170] The results obtained for:
[0171] percent doxorubicin.HCl stability (ion-pair method) and
[0172] pH
[0173] referred to the vials stored in upright position are given
in:
[0174] Table 19 storage at -20.degree. C.
[0175] Tables 20 and 28 storage at +4.degree. C.
[0176] Tables 21 and 29 storage at +8.degree. C.
[0177] Tables 22 and 30 storage at +15.degree. C.
[0178] Tables 23 and 31 storage at +27.degree. C.
[0179] Table 24 storage at 100 and 250 foot candles
[0180] Table 32 storage at 250 foot candles.
[0181] The doxorubicin.HCl assays given in these tables are the
mean of three independent determinations.
[0182] As far as the other parameters checked during stability:
[0183] the clarity of the solution was unchanged at all the checks
carried out at all the storage conditions applied;
[0184] the appearance of the solutions was: a) unchanged at all the
checks carried out on samples stored at 4.degree. C. and 8.degree.
C., b) slightly darkened after: 9 months at 15.degree. C., 3 months
at 27.degree. C., 3 months at 100 and 250 foot candles light;
[0185] the closure system was unchanged at all the checks carried
out at all the storage conditions;
[0186] the sterility was maintained after 18 months at 8.degree.
C.
[0187] The results of the controls carried out on the vials stored
in inverted position do not differ significantly from those on the
vials in upright position.
[0188] The percent doxorubicin.HCl stability values obtained by the
USP HPLC method do not differ significantly from those obtained by
the HPLC ion-pair method given in Tables 19-24.
[0189] The obtained stability data indicate that the tested
doxorubicin.HCl solutions having different pH values within the
range from 2.5 to 3.5 reach the lower limit of acceptance (90% of
nominal concentration) in about 9 and 2-3 months at 15.degree. C.
and, respectively 27.degree. C., but prove stable up to 18 months
at 4.degree. C. and 8.degree. C., i.e. at temperature usually
adopted for the storage of the products under refrigeration.
[0190] In distinct contrast, the doxorubicin.HCl solution obtained
upon reconstitution of the commercial freeze-dried preparate, whose
pH varies between 4.5 and 6, shows a much lower degree of stability
as shown by the fact that it is recommended to discard
reconstituted solutions after only 48 hours storage in refrigerator
according to the leaflet accompanying packages of Adriamycin (i.e.
doxorubicin.HCl) in the United States.
22TABLE 16 Stability studies. Batches tested. Batch Batch No
Characteristics TF/23049 TF/23077 TF/23078 TF/23117 TF/23119 H0001
L0001 M0001 Doxorubicin.multidot.HCl 10 10 10 20 50 10 20 50 per
vial (mg) pH 3.06 2.81 3.50 2.97 3.08 3.15 3.05 3.20 Formulation
No. F16804/IL1 F16804/IL1 F16804/IL1 F16804/IL2 F16804/IL3
F16804/IL4 F16804/IL5 F16804/IL6 Batch size 700 400 400 500 500
2,400 2,300 2,400 No. of vials
[0191]
23TABLE 17 Stability studies. Formulations tested. Composition per
Formulation number vial F16804/IL F16804/IL2 F16804/IL3 F16804/IL4
F16804/IL5 F16804/IL6 Doxorubicin.multidot.HCl 10.0 20 50 10 20 50
mg Hydrochloric acid 2.8-3.5 2.8-3.5 2.8-3.5 2.8-3.5 2.8-3.5
2.8-3.5 q.s. to pH Water q.s. to ml 5.0 10.0 25.0 5.0 10.0 25.0
[0192]
24TABLE 18 Stability studies. Packaging used. Batch No. Packaging
TF/23049 TF/23077 TF/23078 TF/231117 TF/23119 H0001 L0001 M0001
vial glass type I I I I I I I I vial top 8 ml 8 ml 8 ml 14 ml 39 ml
10 ml 14 ml 39 ml capacity stopper chlorobutyl chlorobutyl
chlorobutyl chlorobutyl chlorobutyl chlorobutyl chlorobutyl
chlorobutyl rubber, rubber, rubber, rubber, rubber, rubber, rubber,
rubber, teflon-faced teflon-faced teflon-faced teflon-faced
teflon-faced teflon-faced teflon-faced teflon-faced seal aluminum
aluminum aluminum aluminum aluminum aluminum aluminum aluminum
[0193]
25TABLE 19 Doxorubicin.multidot.HCl 2 mg/ml solution. Stability
data at -20.degree. C. (vials stored upright) acquired up to 3
months. Batch Time - Months Dosage 0 1 3 H0001
doxorubicin.multidot.HCl 100 99.9 99.6 10 mg % stability pH 3.15
3.12 2.98 L0001 doxorubicin.multidot.HCl 100 100.8 99.8 20 mg %
stability pH 3.05 2.84 2.97 M0001 doxorubicin.multidot.HCl 100
100.7 101.0 50 mg % stability pH 3.20 2.96 2.99
[0194]
26TABLE 20 Doxorubicin.multidot.HCl 2 mg/ml solution. Stability
data at 4.degree. C. (vials stored upright) acquired up to 18
months Batch Time Months Dosage 0 1 3 6 9 12 18 TF/23049 * 100 99.9
100.6 98.3 98.2 97.7 96.9 10 mg ** 3.06 3.10 3.09 3.10 3.05 2.97
3.07 TF/23077 * 100 101.7 99.3 97.9 98.0 99.8 10 mg ** 2.81 2.86
2.75 2.65 2.67 2.76 TF/23078 * 100 101.2 98.8 97.8 98.8 96.8 10 mg
** 3.50 3.54 3.49 3.44 3.43 3.54 TF/23117 * 100 96.8 96.6 98.1 98.8
97.5 20 mg ** 2.97 2.98 2.92 2.86 2.95 2.98 TF/23119 * 100 98.6
99.1 98.9 98.4 97.5 50 mg ** 3.08 2.98 2.98 2.89 2.99 3.00 H0001 *
100 97.6 99.2 10 mg ** 3.15 n.d. 3.06 3.22 L000 * 100 98.8 98.4 20
mg ** 3.05 n.d. 2.99 2.94 M0001 * 100 99.7 99.7 50 mg ** 3.20 n.d.
3.00 3.04 * doxorubicin.multidot.HCl % stability ** pH n.d. = not
determined
[0195]
27TABLE 21 Doxorubicin.multidot.HCl 2 mg/ml solution. Stability
data at 8.degree. C. (vials stored upright) acquired up to 18
months Batch Time (Months) Dosage 0 1 2 3 6 9 12 18 TF/23049 * 100
99.7 100.1 96.5 96.1 96.5 95.4 10 mg ** 3.06 3.07 3.09 3.07 3.04
2.96 3.04 TF/23077 * 100 102.1 101.6 97.5 96.6 95.0 10 mg ** 2.81
2.81 2.74 2.65 2.67 2.75 TF/23078 * 100 98.3 97.7 96.5 95.9 98.8 10
mg ** 3.50 3.59 3.47 2.27 3.43 3.51 TF/23117 * 100 95.7 95.8 97.8
96.2 95.5 20 mg ** 2.97 2.97 2.92 2.85 2.96 2.98 TF/23119 * 100
97.6 97.8 96.2 97.3 96.8 50 mg ** 3.08 2.94 2.94 2.87 2.99 3.00
H0001 * 100 98.2 99.4 96.4 96.7 10 mg ** 3.15 3.12 3.16 3.05 3.23
L000 * 100 100.6 99.1 98.1 98.3 20 mg ** 3.05 2.84 2.83 2.97 2.94
M0001 * 100 100.3 100.6 98.7 99.0 50 mg ** 3.20 2.96 2.97 3.01 3.03
* doxorubicin.multidot.HCl % stability ** pH
[0196]
28TABLE 22 Doxorubicin.multidot.HCl 2 mg/ml solution. Stability
data at 15.degree. C. (vials stored upright) acquired up to 12
months Batch Time (Months) Dosage 0 0.5 1 2 3 6 9 12 TF/23049 * 100
97.8 98.9 97.1 92.7 92.9 90.2 10 mg ** 3.06 3.03 3.07 3.10 3.08
3.02 2.95 TF/23077 * 100 100.4 101.9 98.8 94.6 92.7 91.1 10 mg **
2.81 2.81 2.85 2.71 2.63 2.67 2.74 TF/23078 * 100 101.4 98.4 95.3
94.6 91.9 90.7 10 mg ** 3.50 3.51 3.58 3.47 3.38 3.41 3.47 TF/23117
* 100 99.1 96.4 95.2 94.6 90.7 20 mg ** 2.97 2.95 2.95 2.90 2.81
2.95 TF/23119 * 100 97.4 97.1 95.9 92.7 90.6 50 mg ** 3.08 2.99
2.95 2.91 2.87 2.98 H0001 * 100 97.9 97.1 94.8 94.6 10 mg ** 3.15
3.12 3.16 3.06 3.23 L000 * 100 100.5 98.7 96.3 95.5 20 mg ** 3.05
2.85 2.87 2.98 2.96 M0001 * 100 99.4 100.3 97.2 95.6 50 mg ** 3.20
2.96 2.94 3.01 3.04 * doxorubicin.multidot.HCl % stability **
pH
[0197]
29TABLE 23 Doxorubicin.multidot.HCl 2 mg/ml solution. Stability
data at 27.degree. C. (vials stored upright) acquired up to 6
months. Batch Time (Months) Dosage 0 0.5 1 2 3 6 TF/23049 * 100
97.2 95.8 87.9 73.6 10 mg ** 3.06 2.98 3.07 3.08 3.03 TF/23077 *
100 98.5 96.2 86.4 69.2 10 mg ** 2.81 2.80 2.85 2.71 2.64 TF/23078
* 100 101.2 94.5 80.5 71.1 10 mg ** 3.50 3.51 3.58 3.38 3.13
TF/23117 * 100 97.4 93.2 81.9 66.6 20 mg ** 2.97 2.95 2.94 2.88
2.77 TF/23119 * 100 96.0 93.3 85.3 66.8 50 mg ** 3.08 2.97 2.97
2.91 2.82 H0001 * 100 94.5 94.2 86.6 10 mg ** 3.15 3.10 3.09 3.01
L000 * 100 97.2 94.3 89.3 20 mg ** 3.05 2.84 2.85 2.96 M0001 * 100
96.5 93.6 88.1 50 mg ** 3.20 2.95 2.95 2.99 *
doxorubicin.multidot.HCl % stability ** pH
[0198]
30TABLE 24 Doxorubicin.multidot.HCl 2 mg/ml solution. Stability
data at 100 and 250 f.c. (vials stored inverted) acquired up to 3
months. 100 foot-candles 250 foot-candles Batch Time Months Dosage
0 0.5 1 3 0.5 1 2 3 TF/23049 * 100 96.3 95.9 81.3 95.9 94.8 10 mg
** 3.06 3.05 3.05 3.06 2.99 3.04 TF/23077 * 100 98.3 98.1 87.7 97.3
94.5 10 mg ** 2.81 2.79 2.84 2.70 2.79 2.84 TF/23078 * 100 99.6
96.4 88.0 97.8 89.7 10 mg ** 3.50 3.50 3.58 3.39 3.47 3.53 TF/23117
* 100 96.8 96.7 91.7 98.1 94.6 20 mg ** 2.97 2.93 2.95 2.87 2.9
2.93 TF/23119 * 100 96.9 96.7 89.6 96.4 95.0 50 mg ** 3.08 2.96
2.95 2.93 2.96 2.97 H0001 * 100 95.2 93.7 87.8 10 mg ** 3.15 3.10
3.06 2.97 L000 * 100 96.5 93.0 86.5 20 mg ** 3.05 2.84 2.85 2.97
M0001 * 100 97.8 91.5 85.3 50 mg ** 3.20 2.95 2.94 2.99 *
doxorubicin.multidot.HCl % stability ** pH
[0199]
31TABLE 25 Stability studies. Batches tested. Batch Batch No.
Characteristics P0001 Q0001 R0001 Doxorubicin.multidot.HCl 10 20 50
per vial (mg) pH 3.00 3.00 3.00 Formulation No. FI6804/IL7
FI6804/IL8 FI6804/IL9 Batch size 2,400 2,200 2,500 No. of vials
[0200]
32TABLE 26 Stability studies. Formulations tested. Composition per
Formulation number vial F16804/IL7 F16804/IL8 F16804/IL9
Doxorubicin.multidot.HCl 10 20 50 per vial (mg) Hydrochloric acid
2.8-3.5 2.8-3.5 2.8-3.5 q.s. to pH 0.9% sodium chloride 5.0 10.0
25.0 injection q.s. to ml
[0201]
33TABLE 27 Stability studies. Packaging used. Batch No Packaging
P0001 Q0001 R0001 vial glass type I I I vial top 10 ml 14 ml 39 ml
capacity stopper chlorobutyl chlorobutyl chlorobutyl rubber,
rubber, rubber, teflon-faced teflon-faced teflon-faced seal
aluminum aluminum aluminum
[0202]
34TABLE 28 Doxorubicin.multidot.HCl 2 mg/ml solution in Saline for
Injection at pH = 3. Stability data at 4.degree. C. (vials stored
upright) acquired up to 9 and 12 months. Batch Time - Months Dosage
0 3 6 9 12 P0001 doxorubicin.multidot.HCl 100 98.3 98.0 99.2 10 mg
% stability pH 3.00 2.93 2.98 2.90 Q0001 doxorubicin.multidot.HCl
100 97.5 97.0 100.1 20 mg % stability pH 3.01 3.06 3.03 3.00 R0001
doxorubicin.multidot.HCl 100 99.8 100.7 101.2 101.7 50 mg %
stability pH 3.02 3.08 3.15 3.14 3.10
[0203]
35TABLE 29 Doxorubicin.multidot.HCl 2 mg/ml solution in Saline for
Injection at pH = 3. Stability data at 8.degree. C. (vials stored
upright) acquired up to 9 and 12 months. Batch Time - Months Dosage
0 1 2 3 6 9 12 P0001 doxorubicin.multidot.HCl 100 101.0 100.6 97.9
97.4 96.8 10 mg % stability pH 3.00 2.93 2.89 2.91 3.00 2.90 Q0001
doxorubicin.multidot.HCl 100 99.4 99.9 96.8 96.7 95.7 20 mg %
stability pH 3.01 3.02 3.01 3.05 3.02 3.00 R0001
doxorubicin.multidot.HCl 100 99.8 99.8 98.4 98.5 99.5 100.9 50 mg %
stability pH 3.02 3.02 3.09 3.08 3.13 3.13 3.10
[0204]
36TABLE 30 Doxorubicin.multidot.HCl 2 mg/ml solution in Saline for
Injection at pH = 3. Stability data at 15.degree. C. (vials stored
upright) acquired up to 9 and 12 months. Batch Time - Months Dosage
0 1 2 3 6 9 12 P0001 doxorubicin.multidot.HCl 100 100.6 99.9 95.9
94.0 89.1 10 mg % stability pH 3.00 2.93 2.89 2.90 2.99 2.90 Q0001
doxorubicin.multidot.HCl 100 98.6 97.8 95.1 96.4 89.8 20 mg %
stability pH 3.01 3.01 3.01 3.04 3.01 3.00 R0001
doxorubicin.multidot.HCl 100 98.8 97.5 97.6 94.7 96.0 94.5 50 mg %
stability pH 3.02 3.02 3.08 3.08 3.14 3.11 3.10
[0205]
37TABLE 31 Doxorubicin.multidot.HCl 2 mg/ml solution in Saline for
Injection at pH = 3. Stability data at 27.degree. C. (vials stored
upright) acquired up to 3 months. Batch Time - Months Dosage 0 1 2
3 P0001 doxorubicin.multidot.HCl 100 98.3 95.0 84.9 10 mg %
stability pH 3.00 2.93 2.89 2.88 Q0001 doxorubicin.multidot.HCl 100
98.0 93.2 83.8 20 mg % stability pH 3.01 3.01 2.99 3.03 R0001
doxorubicin.multidot.HCl 100 95.6 92.2 88.7 50 mg % stability pH
3.02 3.02 3.06 3.05
[0206]
38TABLE 32 Doxorubicin.multidot.HCl 2 mg/ml solution in Saline for
Injection at pH = 3. Stability data at R.T. + 250 f.c. (vials
stored upright) acquired up to 3 months. Batch Time - Months Dosage
0 1 2 3 P0001 doxorubicin.multidot.HCl 100 89.6 86.5 70.3 10 mg %
stability pH 3.00 2.92 2.86 2.84 Q0001 doxorubicin.multidot.HCl 100
91.1 84.5 72.7 20 mg % stability pH 3.01 2.99 2.97 2.98 R0001
doxorubicin.multidot.HCl 100 96.0 91.4 86.6 50 mg % stability pH
3.02 3.01 3.04 3.02
[0207]
39TABLE 33 Stability Data of Doxorubicin Solution, 2 mg/ml and a pH
of 3.0 at 45.degree. C. Stabilizing Agent % Initial and Its
Concentration 1 Wk 2 Wk 4 Wk 8 Wk Water 87.8 75.9 53.8 25.5 5%
Dextrose 91.1 82.3 65.6 38.8 5% Galactose 91.5 86.1 64.3 -- 5%
Fructose 91.9 80.6 64.1 -- 4% .alpha.-L(-)-Fucose 91.2 81.9 63.8 --
4% .alpha.-D(+)-Fucose 91.8 81.9 63.3 -- 1% Lactose 91.3 81.7 64.5
34.8 4% Dextran, MW 9,000 90.5 81.5 -- -- 4% Dextran, MW 506,000
92.0 84.0 -- -- 4% .alpha.-Cyclodextrin 91.7 84.3 -- -- 4%
.beta.-Cyclodextrin 92.1 84.1 -- -- 4% .gamma.-Cyclodextrin 94.3
89.0 -- -- 5% Mannitol 90.7 81.4 65.8 41.1 5% Sorbitol 91.4 83.0
67.2 42.5 0.5% Thioglycerol 90.8 83.2 63.5 -- 5% Inositol 91.7 84.9
-- -- 5% Ethanol 92.2 85.6 -- -- 10% Glycerol 92.2 83.4 65.5 --
Note: The same stabilizing effect may be seen for the above agents
at lower concentrations, e.g., lower by up to 25-50 wt. %.
[0208] The invention now being fully described, it will be apparent
to one of ordinary skill in the art that many changes and
modifications can be made thereto without departing from the spirit
or scope of the invention as set forth herein.
* * * * *