U.S. patent application number 09/192933 was filed with the patent office on 2002-01-24 for stable amorphous amifostine compositions and dosage form.
Invention is credited to STOGNIEW, MARTIN, ZADEI, JAVAD.
Application Number | 20020010357 09/192933 |
Document ID | / |
Family ID | 22711626 |
Filed Date | 2002-01-24 |
United States Patent
Application |
20020010357 |
Kind Code |
A1 |
STOGNIEW, MARTIN ; et
al. |
January 24, 2002 |
STABLE AMORPHOUS AMIFOSTINE COMPOSITIONS AND DOSAGE FORM
Abstract
The present invention relates to a sterile, stable dosage forms
suitable for reconstitution and parenteral administration to a
patient, said dosage form comprising an amorphous aminoalkyl
dihydrogen phosphorothioate, and of amifostine in particular. The
invention further relates to a method of preparing such a dosage
form, which typically exhibits enhanced thermal stability as
compared to existing vacuum dried amorphous amifostine.
Inventors: |
STOGNIEW, MARTIN; (BLUE
BELL, PA) ; ZADEI, JAVAD; (WEST CHESTER, PA) |
Correspondence
Address: |
PENNIE & EDMONDS LLP
1667 K STREET NW
WASHINGTON
DC
20006
|
Family ID: |
22711626 |
Appl. No.: |
09/192933 |
Filed: |
November 16, 1998 |
Current U.S.
Class: |
558/146 |
Current CPC
Class: |
A61K 31/66 20130101;
A61P 39/00 20180101; A61K 9/19 20130101; A61P 35/00 20180101; A61K
47/183 20130101; A61K 31/661 20130101; A61K 9/0019 20130101 |
Class at
Publication: |
558/146 |
International
Class: |
A61K 031/47; C07F
009/02 |
Claims
What is claimed is:
1. A dosage form of amifostine which comprises thermally-stable,
sterile, amorphous amifostine which is suitable for reconstitution
with a pharmaceutically acceptable vehicle into an injectable
particulate-free drug product for parenteral administration to a
subject.
2. A dosage form of amifostine suitable for reconstitution with a
pharmaceutically acceptable vehicle into an injectable
particulate-free drug product for parenteral administration to a
subject which comprises thermally-stable, sterile, amorphous
amifostine.
3. A thermally-stable, amorphous, sterile dosage form comprising
amifostine.
4. A thermally-stable amorphous dosage form comprising sterile
amifostine suitable for reconstitution with a pharmaceutically
acceptable vehicle into an injectable particulate-free drug product
for parenteral administration to a subject.
5. The dosage form of claim 1 further comprising a stabilizer.
6. The dosage form of claim 5 wherein the stabilizer is
substantially amorphous.
7. The amorphous dosage form of claim 5 wherein the stabilizer is
an amide or an amino acid.
8. The dosage form of claim 7 wherein the amide or amino acid is
selected from the group consisting of nicotinamide, derivatives of
nicotinamide, nicotinic acid, derivatives of nicotinic acid,
glycine, alanine, valine, leucine, isoleucine, phenylalanine,
tryptophan, asparagine, and mixtures thereof.
9. The dosage form of claim 5 wherein the molar ratio of stabilizer
to amifostine is between about 0.05 and about 5.0.
10. The dosage form of claim 9 wherein the molar ratio of
stabilizer to amifostine is between about 0.1 and about 1.0.
11. The dosage form of claim 10 wherein the molar ratio of
stabilizer to amifostine is between about 0.2 and about 0.5.
12. The dosage form of claims 1 or 5 further comprising an
excipient.
13. The dosage form of claim 12 wherein the excipient is selected
from the group consisting of sodium chloride, citric acid, tartaric
acid, gelatin and carbohydrates such as dextrose, sucrose,
sorbitol, inositol, dextran, mannitol, EDTA, and DTPA.
14. The dosage form of claim 1, 5 or 12, said dosage form having a
water content of between about 0 weight percent and about 20 weight
percent.
15. The dosage form of claim 14, said dosage form having a water
content of between about 0 weight percent and about 15 weight
percent.
16. The dosage form of claim 15, said dosage form having a water
content of between about 0.5 weight percent and about 6 weight
percent.
17. The dosage form of claims 1 wherein said dosage form forms less
than about 5 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
40.degree. C. for about one week, said weight percent based upon
total weight of amifostine.
18. The dosage form of claim 17 wherein said dosage form forms less
than about 4 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
40.degree. C. for about one week, said weight percent based upon
total weight of amifostine.
19. The dosage form of claim 18 wherein said dosage form forms less
than about 3 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in an inert gas filled vial and maintained at a
temperature of about 40.degree. C. for about one week, said weight
percent based upon total weight of amifostine.
20. The dosage form of claims 1 wherein said dosage form forms less
than about 5 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
25.degree. C. for about one month, said weight percent based upon
total weight of amifostine.
21. The dosage form of claim 20 wherein said dosage form forms less
than about 4 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
25.degree. C. for about one month, said weight percent based upon
total weight of amifostine.
22. The dosage form of claim 21 wherein said dosage form forms less
than about 3 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
25.degree. C. for about one month, said weight percent based upon
total weight of amifostine.
23. The dosage form of claim 22 wherein said dosage form forms less
than about 2 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
25.degree. C. for about one month, said weight percent based upon
total weight of amifostine.
24. The dosage form of claim 1 wherein said dosage form forms less
than about 3 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
5.degree. C. for about two years, said weight percent based upon
total weight of amifostine.
25. The dosage form of claim 24 wherein said dosage form forms less
than about 2 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
5.degree. C. for about two years, said weight percent based upon
total weight of amifostine.
26. The dosage form of claim 25 wherein said dosage form forms less
than about 1 weight percent of 2-[(3-aminopropyl)amino]ethane thiol
when sealed in a vial and maintained at a temperature of about
5.degree. C. for about two years, said weight percent based upon
total weight of amifostine.
27. A room temperature stable dosage form of amifostine comprising
sterile, amorphous amifostine which is suitable for reconstitution
with a pharmaceutically acceptable vehicle into an injectable
particulate-free drug product for parenteral administration to a
subject.
28. A room temperature stable amorphous dosage form comprising
sterile, amorphous amifostine which is suitable for reconstitution
with a pharmaceutically acceptable vehicle into an injectable
particulate-free drug product for parenteral administration to a
subject.
29. A refrigerated stable dosage form comprising sterile, amorphous
amifostine, wherein said dosage form forms less than about 3 weight
percent of 2-[(3-aminopropyl)amino]ethane thiol when sealed in a
vial and maintained at a temperature of about 5.degree. C. for
about two years, said weight percent based upon total weight of
amifostine.
30. A dosage form of amifostine consisting essentially of
thermally-stable, sterile, amorphous amifostine.
31. A dosage form of amifostine consisting of thermally-stable,
sterile, amorphous amifostine.
32. A thermally-stable dosage form comprising sterile amorphous
amifostine and nicotinamide in an amifostine to nicotinamide weight
ratio of between about 4:1 and about 10:1.
33. The dosage form of claim 32 wherein the amifostine to
nicotinamide weight ratio is about 8:1.
34. The dosage form of claim 27 or 29 further comprising a
stabilizer.
35. The dosage form of claim 27 or 29 further comprising an
excipient.
36. The dosage form of claim 27 or 29, said dosage form having a
water content of between about 0 weight percent and about 20 weight
percent.
37. A dosage form of claim 1, 2, 3, 27, 28 or 28, wherein said
dosage form is substantially amorphous.
38. A dosage form of claim 1, 2, 3, 27, 28 or 28, wherein said
dosage form is at least about 80% amorphous by weight.
39. A dosage form of claim 1, 2, 3, 27, 28 or 28, wherein said
dosage form is at least about 90% amorphous by weight.
40. A process for the preparation of a thermally-stable dosage form
of amorphous amifostine comprising lyophilizing a sterile aqueous
solution of amifostine wherein said lyophilization comprises: a
freezing cycle having a first average temperature of between about
-70.degree. C. and about -10.degree. C. and lasting for a first
period of time of between about 1 hour and about 15 hours; and a
primary drying cycle having a second average temperature of between
about -45.degree. C. and about 10.degree. C. and lasting for a
second period of time of between about 8 hours and about 100
hours.
41. The process of claim 40 wherein the aqueous solution is
substantially free of alcohol.
42. The process of claim 40 wherein the aqueous solution has a pH
of between about 6 and about 9.
43. The process of claim 40 further comprising dissolving a
stabilizer in the aqueous solution prior to lyophilization.
44. The process of claim 40 further comprising dissolving an
excipient in the aqueous solution prior to lyophilization.
45. The process of claim 40 further comprising mixing the product
formed upon lyophilization with a stabilizer.
46. The process of claim 40 further comprising mixing the product
formed upon lyophilization with an excipient.
47. The process of claim 43 or 45 wherein the stabilizer is an
amide, amino acid, or pharmaceutically acceptable base.
48. The process of claim 47 wherein the amide or amino acid is
selected from the group consisting of nicotinamide, derivatives of
nicotinamide, nicotinic acid, derivatives of nicotinic acid,
glycine, alanine, valine, leucine, isoleucine, phenylalanine,
tryptophan, asparagine, and mixtures thereof.
49. The process of claim 47 wherein the molar ratio of stabilizer
to bulk amifostine is between about 0.05 and about 5.0.
50. The process of claim 49 wherein the molar ratio of stabilizer
to bulk amifostine is between about 0.1 and about 1.0.
51. The process of claim 50 wherein the molar ratio of stabilizer
to bulk amifostine is between about 0.2 and about 0.5.
52. The process of claim 40 wherein the first average temperature
is between about -65.degree. C. and about -30.degree. C.
53. The process of claim 52 wherein the first average temperature
is between about -55.degree. C. and about -40.degree. C.
54. The process of claim 40 wherein the first time is between about
2 hours and about 10 hours.
55. The process of claim 54 wherein the first time is between about
5 hours and about 8 hours.
56. The process of claim 40 wherein the second average temperature
is between about -35.degree. C. and about 0.degree. C.
57. The process of claim 56 wherein the second average temperature
is between about -30.degree. C. and about -10.degree. C.
58. The process of claim 40 wherein the second time is between
about 16 hours and about 86 hours.
59. The process of claim 58 wherein the second time is between
about 24 hours and about 72 hours.
60. The process of claim 40 wherein lyophilizing the solution
further comprises a secondary drying step having a third average
temperature of between about -25.degree. C. and about 40.degree. C.
and a third average time of between about 0.5 hours and about 48
hours.
61. The process of claim 60 wherein the third average temperature
is between about -15.degree. C. and about 40.degree. C.
62. The process of claim 61 wherein the third average temperature
is between about 0.degree. C. and about 35.degree. C.
63. The process of claim 60 wherein the third time is between about
2 hours and about 36 hours.
64. The process of claim 63 wherein the third time is between about
6 hours and about 24 hours.
65. The process of claim 40 further comprising stoppering the vials
with desiccating stoppers after lyophilizing the solution.
66. The process of claim 40 wherein the aqueous solution is
lyophilized under conditions such that the dosage form of
amifostine has a water content of between about 0 weight percent
and about 20 weight percent.
67. The process of claim 66 wherein the aqueous solution is
lyophilized under conditions such that the dosage form of
amifostine has a water content of between about 0 weight percent
and about 15 weight percent.
68. The process of claim 67 wherein the aqueous solution is
lyophilized under conditions such that the dosage form of
amifostine has a water content of between about 0.5 weight percent
and about 6 weight percent.
69. A thermally-stable dosage form of amorphous amifostine formed
by the process of claim 40.
Description
FIELD OF INVENTION
[0001] The present invention relates to thermally-stable dosage
forms of S-2-(3-aminopropylamino)ethyl dihydrogen phosphorothioate
(amifostine), and to methods of making same. The reconstituted
dosage forms of the invention are suitable for administration to
humans, for example, as a radio- or chemoprotectant agent.
BACKGROUND OF THE INVENTION
[0002] The compound S-2-(3-aminopropylamino)ethyl dihydrogen
phosphorothioate (which is also known as amifostine, ethiofos,
Ethyol.RTM., NSC 296961, and WR-2721 and which will hereinafter be
referred to as "amifostine") and other aminothiol compounds are
disclosed in U.S. Pat. No. 3,892,824. These compounds were
originally developed as antiradiation agents (radio-protectants),
in particular to be used prior to exposure to x-ray or nuclear
radiation, to protect against the harmful effects of such exposure
which may be encountered during military conflicts.
[0003] In addition to its utility as a military antiradiation
agent, amifostine has demonstrated excellent utility as a
non-military radioprotectant and chemoprotectant, i.e., as a
protectant administered prior to therapy to reduce the undesirable
adverse effects which arise during the use of chemotherapy and
radiation therapy in the treatment of cancer. Nygaard et al., eds,
Radioprotectors and Anticarcinogens, Academic Press, Inc., New
York, pp. 73-85 (1983); Grdina et al., Carcinogenesis (London)
6:929-931 (1985). In addition, these compounds have been reported
to afford protection against the adverse effects of
chemotherapeutic agents, for example, alkylating agents such as
cisplatin, when administered before or concurrently with the
chemotherapeutic agent. Jordan et al., Exp. Mol. Pathol. 36:297
(1982); Doz et al., Cancer Chemother. Pharmacol. 28:308 (1991).
Similarly, it has been reported that amifostine has been used
experimentally prior to therapy to protect HIV-infected patients
(AIDS) from the harmful side effects of 3'-azido-3'-deoxythymidine
(AZT) therapy. International Published Application WO 90/14007,
published Nov. 29, 1990. Amifostine and its derivatives have been
shown to exert these reported protective effects without affecting
the beneficial properties of the administered therapeutic agents.
This is, in the case of chemotherapy, believed to be due to the
selective uptake of the protective thiol and other metabolites into
normal tissue. Yuhas, Cancer Res. 40:1519-1524 (1980); Yuhas,
Cancer Treat. Rep. 63:971-976 (1979).
[0004] Amifostine and related aminothiol compounds have also been
shown to stimulate bone marrow growth. See, eg., International
Published Application WO 96/25045 published Aug. 22, 1996; and List
et al., Proc. Am. Soc. Clin. Oncol. 15:449 [1403] [Abstract].
Currently, amifostine is in Phase II clinical trials as a bone
marrow stimulant in patients suffering from myelodysplastic
syndrome. Pre-exposure with aminothiol compounds is capable of
causing the bone marrow function to more rapidly recover following
chemotherapy. List et al., Semin. Oncol. 23 (4) Supp. 8:58-63
(1996).
[0005] Presently, amifostine is indicated to reduce the cumulative
renal toxicity associated with repeated administration of cisplatin
in patients with advanced ovarian or non-small cell lung cancer.
Physicians' Desk Reference 52.sup.nd ed., p. 500-502 (1998).
[0006] Amifostine is considered a prodrug. Amifostine is
metabolized to the cytoprotective free thiol by dephosphorylation
catalyzed by alkaline phosphatase. See, e.g., Ryan, S. V., et al.,
J. Clin. Pharm. 36(4):365-373 (1996). Amifostine exerts protective
effects without significantly affecting the beneficial properties
of the administered therapeutic agents largely because of the
selective uptake of the thiol into normal tissue.
[0007] In its most common use, amifostine is administered
parenterally, including by bolus injection and intravenous
infusion. Amifostine is also being developed for subcutaneous
administration. Since these routes circumvent the protective
barriers of the human body, exceptional purity of the dosage form
must be achieved. Because the dosage form must be free of
microoganisms and insoluble particulates, the process used in
preparing it must embody Good Manufacturing Practices ("GMP") that
will produce and maintain the required quality of the product in
terms of sterility and therapeutic effectiveness. Sterility is
especially important in the treatment of cancer and AIDS patients,
because in many instances they are already immuno-compromised and
therefor highly susceptible to infections.
[0008] The amifostine bulk drug (which is distinct from the dosage
form) can exist as a bulk crystalline trihydrate which is believed
to be relatively temperature stable. Such a crystalline form is
described by Karle et al., Acta Cryst. C44:135-138 (1988). The bulk
drug, however, is not sterile, and so cannot be reconstituted into
a pharmaceutical product suitable for parenteral administration to
humans.
[0009] Several methods of sterilizing bulk drugs are described in
Remington's Pharmaceutical Sciences, 18.sup.th ed. (1990). These
include, for example, steam sterilization, wherein a drug is
exposed to high pressure steam at a minimum temperature of
121.degree. C. This and other methods that require heating,
however, cannot effectively be used to sterilize bulk crystalline
amifostine. This is because crystalline amifostine loses water at
about 70.degree. C. to about 75.degree. C. The loss of water
facilitates degradation by a hydrolysis reaction that forms
phosphoric acid and 2-[(3-aminopropyl)amino]ethane thiol. See,
e.g., Risley, J. M. and Van Etten, R. L., Biochem. Pharmacol.
35:1453-1458 (1986). Amifostine has thus been sterilized by
dissolving it in an aqueous solution which is then sterilized by
membrane filtration. Substantial practical problems related to the
packaging of bulk, solid amifostine using the so-called "dry
filling" or "powder filling" method were thus avoided. These
problems include producing sterile amifostine bulk, the difficulty
in the manual manipulation of powders, the need to mill the powders
to acceptable particle size and flowability, difficulty in
maintaining particle-free, aseptic conditions, and the difficulty
in supplying the precise dosages into individual vials.
[0010] In solution, however, amifostine is again susceptible to
degradation by hydrolysis. For this reason, a prior process has
sterilized an amifostine solution by filtration, and then
lyophilized (freeze-dried) the amifostine solution under the
following conditions: about 5 ml of a solution of 100 mg/ml
amifostine and 100 mg/ml mannitol placed in vial was loaded into a
freeze-drier at 0.degree. C. and then solidified at -45.degree. C.
The vials were kept at -45.degree. C. for 2 hours, after which time
the freeze drying chamber was evacuated to 100 .mu.m Hg. The self
temperature was then raised to 0.degree. C. over 12 hours and then
maintained at 0.degree. C. for 2 hours. The shelf temperature was
finally raised to 25.degree. C., at which temperature the vials
were kept for 24 hours. This method produced a sterile amorphous
form of amifostine suitable for parenteral administration to a
patient (which is hereinafter referred to as "Amorphous Amifostine
I"). Physicians' Desk Reference, 51.sup.st ed. (1997) p.485-486.
This method also allowed easy production of vials containing
predetermined amounts of sterile, lyophilized amifostine.
[0011] Unfortunately, Amorphous Amifostine I was thermally unstable
and hydrolyzed over time at temperatures above 0.degree. C. For
example, a typical sample of Amorphous Amifostine I kept at
25.degree. C. for about one month formed about 6 to 7 weight
percent of the degradation product 2-[(3-aminopropyl)amino]ethane
thiol. This was the state of the art when the amorphous form of
amifostine was approved by regulatory authorities as having a shelf
life of 24 months when stored at temperatures of between about
2.degree. C. and 8.degree. C. See e.g., Ethyol.RTM. European
Summary of Product Characteristics (1997) ("Ethyo.RTM. SmPC"). In
fact, the Amorphous Amifostine I dosage form was generally
packaged, shipped, and stored at temperatures below about 8.degree.
C. Further, in the United States, the amorphous form of amifostine
was approved by the Food and Drug Administration as having a shelf
life of 15 months when stored at temperatures of between about
2.degree. C. and 8.degree. C.
[0012] Temperature instability imposes upon manufacturers and users
packaging, storage and shipping requirements that increase the cost
of handling the drug and make its use by hospitals and clinics more
difficult, and may even prevent its use in developing countries
which often lack temperature controlled storage maintenance
capabilities.
[0013] In recognition of the need for a dosage form of amifostine
that is more thermally-stable than Amorphous Amifostine I, research
has been directed at finding new stable and sterile dosage forms of
amifostine. For example, Jahansouz and coworkers studied the
stability of amifostine and suggested that temperature instability
of Amorphous Amifostine I is related to its degree of water
content. Jahansouz, H., et al. Pharm. Res. 7(9):S195 (1990)
[Abstract].
[0014] Recently, instead of attempting to increase the stability of
dosage forms of amorphous amifostine such as Amorphous Amifostine
I, researchers focused on creating stable and sterile crystalline
dosage forms. For example, it was reported that amifostine
"undergoes solid state transitions from the amorphous to
crystalline phases by treatment of the freeze-dried products with
varying humidity." Further, the stability of rehydrated forms of
amifostine was allegedly increased by the addition of certain
excipients. Zadeii, J. M., et al, Pharm. Res. 8(10):S172 (1991)
[Abstract]. This abstract does not, however, describe the
rehydration process in any detail, nor does it describe the degree
of stabilization provided by the two excipients it discloses. In
addition, the abstract provides no detail as to how and when
particular excipients should be combined with amifostine to yield
the alleged more stable rehydrated forms (i.e., whether the
excipients were combined with amifostine prior to, during, or after
its rehydration), or whether the process should differ for
different excipients. Finally, a novel dosage form of sterile,
thermally-stable crystalline amifostine was discovered. U.S. Pat.
Nos. 5,424,471 and 5,591,731.
[0015] Presently, that sterile crystalline dosage form of
amifostine is sold under the trade name Ethyol.RTM.. Physicians'
Desk Reference, 52.sup.nd ed. (1998) p. 500-502. A crystal
structure and preparation of a dosage form, which exhibits greater
thermal stability than Amorphous Amifostine I, is described by U.S.
Pat. Nos. 5,424,471 and 5,591,731. Generally, this thermally-stable
crystalline dosage form is made by dissolving bulk amifostine in a
solvent solution (that allows precipitation of amifostine upon
cooling below 0.degree. C.), followed by sterilization,
precipitation and lyophilization of the compound. Because the
crystallinity, and hence the stability, of this amifostine dosage
form requires three water molecules per molecule, the
lyophilization used to provide the final product must be carefully
controlled to ensure the required degree of hydration. Once made,
the stable crystalline dosage form may be stored at room
temperature with minimal degradation: it exhibits less than 2
weight percent degradation when kept at 40.degree. C. for 14 days.
U.S. Pat. No. 5,591,731.
[0016] Although newly developed sterile crystalline dosage forms of
amifostine reduce or eliminate many of the problems associated with
Amorphous Amifostine I, production of such can be costly, difficult
and/or dangerous. For example, the production of crystalline
Ethyol.RTM. requires the use of explosive solvents which must be
handled with care and strictly isolated from oxygen. Such solvents
also contain impurities not typically found in water that must be
removed to ensure that the dosage form is suitable for
reconstitution and administration to a patient. For example,
ethanol often contains acetone, methanol, H.sub.2SO.sub.4,
MnO.sub.4, and other toxic residues. See, e.g., Aldrich Catalog
(1998-1999), pages 746-747. The production of crystalline
amifostine dosage forms further requires long lyophilization drying
times in order to ensure precise amifostine hydration.
[0017] In view of the above-described and other difficulties
associated with the production of sterile crystalline dosage forms
of amifostine, there remains a need for a stable, sterile amorphous
dosage form, as well as an efficient and inexpensive means of its
production.
[0018] The invention also encompasses non-sterile and/or bulk
thermally-stable amorphous amifostine. To applicants' knowledge, to
date an amorphous thermally-stable amifostine has not been
previously disclosed. In preferred embodiments, the invention
encompasses thermally-stable dosage forms of amifostine.
SUMMARY OF THE INVENTION
[0019] The present invention encompasses non-sterile and/or bulk
thermally-stable amorphous amifostine. To applicants' knowledge,
amorphous thermally-stable amifostine has not been previously
disclosed. In preferred embodiments, the invention encompasses
thermally-stable dosage forms of amifostine.
[0020] The invention encompasses the following thermally-stable and
sterile dosage forms: a dosage form which comprises amorphous
amifostine; an amorphous dosage form which comprises amifostine;
and a dosage form that is suitable for reconstitution with a
pharmaceutically acceptable vehicle into an injectable
particulate-free drug product suitable for parenteral
administration to a subject, which comprises amorphous
amifostine.
[0021] The invention also encompasses a room temperature stable
dosage form comprising sterile amorphous amifostine.
[0022] The invention also includes a room temperature stable
amorphous dosage form comprising sterile, amorphous amifostine.
[0023] This invention is further directed to a refrigerated stable
dosage form comprising sterile amorphous amifostine.
[0024] The dosage forms of this invention may comprise a
stabilizer. It is preferred that the molar ratio of stabilizer to
amifostine be between about 0.05 and about 5.0, more preferably
between about 0.1 and about 1.0, and most preferably between about
0.2 and about 0.5. Preferred stabilizers are compounds having amide
or amino acid moieties. A most preferred stabilizer is
nicotinamide.
[0025] The dosage form of the present invention may also comprise
an excipient. Suitable excipients include sodium chloride, citric
acid, tartaric acid, gelatin and carbohydrates such as dextrose,
sucrose, sorbitol, inositol, dextran, and mannitol,
ethylenediamine-tetraacetic acid (EDTA), sodium deoxycholate,
sodium taurocholate and dietylenetriamine-pentaacetic acid (DTPA).
Finally, the dosage forms may also contain up to about 20 percent
water without significant loss of thermal stability.
[0026] The present invention is further directed to a process of
making a thermally-stable and sterile amorphous dosage form of
amifostine, and to the products of that process.
DEFINITIONS
[0027] As used herein, the terms "bulk amifostine" or "bulk drug"
mean a form of amifostine which is used in preparing dosage forms,
but is not suitable for parenteral administration to a patient. The
term encompasses amifostine as disclosed by U.S. Pat. No.
3,892,824, including, but not limited to, mono-, di- and trihydrate
forms of amifostine.
[0028] As used herein, the term "crystalline," when used to
describe a component or product, means that the component or
product is crystalline as determined by x-ray diffraction. See,
e.g., Remington's Pharmaceutical Sciences, 18.sup.th ed. page 173;
The United States Pharmacopeia, 23.sup.rd ed. (1995) pages
1843-1844.
[0029] As used herein, the term "amorphous" means that the
component or product in question is not crystalline as determined
by x-ray diffraction.
[0030] It should be recognized that a multi-component product,
e.g., a product having amifostine and excipients and/or
stabilizers, may have crystalline and amorphous components, e.g.,
amorphous amifostine and crystalline excipients and/or
stabilizers.
[0031] As used herein, the terms "degradation" and "decomposition"
when used in connection with amifostine refer to the production of
2-[(3-aminopropyl)amino]ethane thiol (WR1065) and phosphoric acid
from amifostine.
[0032] As used herein, the term "Amorphous Amifostine I" means a
dosage form of amifostine that contained amorphous amifostine and
which gave an average of about 3.5% degradation product (i.e.,
2-[(3-aminopropyl)amino]- ethane thiol) at about 5.degree. C. for 2
years. An example of the preparation of Amorphous Amifostine I is
described in the Background of the Invention section above.
[0033] As used herein, the term "Amorphous Amifostine II" refers to
a dosage form of this invention which is suitable for
reconstitution and parenteral administration to a patient. In
particular, Amorphous Amifostine II refers to a dosage form of
amorphous amifostine as described herein.
[0034] As used herein, the terms "stability" and "thermal
stability" refer to the ability of a composition to withstand
degradation or decomposition when kept at a particular temperature
for a specified period of time, preferably under inert atmosphere.
Appropriate means of determining stability are defined herein.
[0035] As used herein, the term "thermally-stable" means more
stable than Amorphous Amifostine I (i.e., the previously commercial
amorphous dosage form described in the Background of the Invention
section above).
[0036] As used herein, the term "stabilizer" means a compound or
mixture of compounds that when mixed in a sufficient amount with
bulk amifostine, and used in the production of an amifostine dosage
form, increases the thermal-stability of the dosage form, for
example by reducing the amount of 2-[(3-aminopropyl)amino]ethane
thiol formed by it over time.
[0037] As used herein, a range of numbers or values defined by the
use of the term "between" includes the particular numbers or values
used to describe that range.
[0038] As used herein, the term "month" means a period of time of
between about 26 days and about 33 days.
[0039] As used herein, the term "weight percent" when used to
describe the amount of degradation product in a dosage form means
the weight of degradation product based upon the weight of
amifostine originally in that dosage form.
[0040] As known to those skilled in the art, accelerated studies
can be used for the determination of stabilities and these
temperatures. See e.g., L. Lachman, et al. The Theory and Practice
of Industrial Pharmacy pages 766-67 (1986). The less a compound or
mixture of compounds degrades when kept at a particular temperature
for a particular time, the more thermally-stable it is.
[0041] As used herein, the terms "refrigeration" or "refrigeration
conditions" mean the maintenance of a temperature of between about
1.degree. C. and about 8.degree. C.
[0042] As used herein, the terms "freeze" or "freezer conditions"
mean the maintenance of a temperature of below about 0.degree. C.,
particularly between about 0.degree. C. and about -20.degree.
C.
[0043] As used herein, the term "particulate-free" means a solution
that is sterile, suitable for bolus injection, intravenous
infusion, or subcutaneous administration, and meets the particulate
matter test described in U.S. Pharmacopia, page 1816 (23.sup.rd
ed., 1995). Specifically, a solution is particulate-free if the
average number of particles having a diameter of greater than 10
.mu.m in the solution does not exceed 6000 per container and the
average number of particles having a diameter of greater than 25
.mu.m in the solution does not exceed 600 per container.
FIGURES
[0044] FIG. 1. Powder x-ray diffraction pattern of crystalline
amifostine prepared as described by U.S. Pat. No. 5,591,731.
[0045] FIG. 2. Powder x-ray diffraction pattern of an amorphous
amifostine dosage form of the present invention wherein
nicotinamide is used as a stabilizer (8:1 amifostine to
nicotinamide weight ratio). This diffraction pattern contains
clearly discernable peaks where 2 .theta. is approximately equal to
the following: 7.9, 13.0, 16.1, 27.4 and 32.8.
[0046] FIG. 3. Powder x-ray diffraction pattern of
nicotinamide.
[0047] FIG. 4. Comparison of the powder x-ray diffraction patterns
of (A) crystalline amifostine prepared as described by U.S. Pat.
No. 5,591,731; and (B) that of an amorphous amifostine dosage form
of the present invention wherein nicotinamide is used as a
stabilizer (8:1 amifostine to nicotinamide weight ratio).
DETAILED DESCRIPTION OF THE INVENTION
[0048] Prior to the present invention, the available
pharmaceutically useful dosage form of amorphous amifostine
(Amorphous Amifostine I) was thermally unstable, and would, for
example, typically yield about 6 weight percent degradation product
(i.e., 2-[(3-aminopropyl)amino]ethane thiol) when kept at
25.degree. C. for one month. Because of its temperature
instability, Amorphous Amifostine I was typically shipped and
stored at low temperatures (generally 8.degree. C., e.g., about
5.+-.3.degree. C.) in order to prevent its degradation. Now, the
present invention provides a dosage form of amorphous amifostine
(Amorphous Amifostine II) that is unexpectedly more
thermally-stable than Amorphous Amifostine I.
[0049] The present invention can be applied to other aminoalkyl
dihydrogen phosphorothiates, and thus provides a more stable dosage
forms of these aminoalkyl dihydrogen phosphorothioates. Aminoalkyl
dihydrogen phosphorothioates suitable for use in the present
invention include, but are not limited to,
S-2-(3-aminopropylamino)ethyl dihydrogen phosphorothioate
(amifostine), S-2-(3-methylaminopropylamino)ethyl dihydrogen
phosphorothioate (WR-3689), S-2-(3-ethylaminopropylamino)ethyl
dihydrogen phosphorothioate,
S-2-(3-aminopropylamino)-2-methylpropyl dihydrogen
phosphorothioate, S-2-(2-aminoethylamino)-2-ethyl dihydrogen
phosphorothioate, S-2-(4-aminobutylamino)-2-ethyl dihydrogen
phosphorothioate, S-2-(5-aminopentylamino)-2-ethyl dihydrogen
phosphorothioate, S-2-(6-aminohexylamino)-2-ethyl dihydrogen
phosphorothioate, S-2-(2-methylaminoethylamino)-2-ethyl dihydrogen
phosphorothioate, S-2-(3-methylaminopropylamino)-2-ethyl dihydrogen
phosphorothioate,
1-propanethiol-3-[[3-(methylamino)propyl]amino]-dihydro- gen
phosphothiorate) (WR-15327) and its free thiol form (WR-151326),
and the disulfide
[2-[(aminopropyl)amino]ethanthiol]-N,N'-dithioidi-2,1-ethan-
ediyl)bis-1,3-propanediamine) (WR-33278). In a preferred embodiment
of the invention, the aminoalkyl dihydrogen phosphorothioate is
amifostine.
[0050] In particular, the present invention provides a
pharmaceutical dosage form of amorphous amifostine suitable for
reconstitution and parenteral administration to a patient that can
be conveniently shipped, stored and handled at room temperature and
refrigerated temperatures with less product degradation than
Amorphous Amifostine I. The dosage forms of this invention (e.g.,
Amorphous Amifostine II) thus provide a solution to a long felt
need: the invention allows relatively low cost amifostine dosage
forms to be more easily shipped to, and more easily stored in,
hospitals and clinics which lack temperature controlled storage and
maintenance capabilities required for Amorphous Amifostine I.
[0051] The present invention is based in part upon the unexpected
discovery that particular amounts of certain compounds when mixed
with amifostine can be used to produce amorphous dosage forms with
reduced rates of decomposition. These certain compounds are herein
referred to as "stabilizers." Without being limited by theory, it
is believed that these stabilizers form van der Waals or a similar
weak intermolecular interactions with the amifostine sulfur atom.
These interactions may prevent the activation (e.g., protonation)
of the sulfur atom. These stabilizers may also inhibit the
interaction of water with, and prevent nucleophilic attack upon,
the amifostine phosphorous atom.
[0052] Because the presence of some stabilizers have been found to
interfere with the measurement of the amifostine content of dosage
form samples, the stability of a dosage form comprising a
stabilizer is preferably determined by measuring the formation of
amifostine degradation or decomposition products as a function of
temperature and time. Such products include, but are not limited
to, 2-[(3-aminopropyl)amino]ethane thiol (the primary degradation
product of amifostine) and phosphoric acid.
[0053] It has also been discovered that the stability of the dosage
forms of amorphous amifostine is dependent upon the freeze drying
process used for its preparation, and that the simple mixing of
amifostine and a stabilizer does not necessarily yield a more
stable dosage form. Even more unexpected is the discovery that,
contrary to the teaching of the prior art, dosage forms of
amorphous amifostine comprising different amounts of water may be
thermally-stable.
[0054] Consequently, this invention is directed to thermally-stable
dosage form of amifostine. In particular, it is directed to a
dosage form that exhibits less than 5 weight percent degradation
when maintained at 40.degree. C. for one week, said weight percent
based upon total weight of amifostine; preferably less than 4
weight percent degradation when maintained at 40.degree. C. for one
week, said weight percent based upon total weight of amifostine;
and more preferably less than 3 weight percent degradation when
maintained at 40.degree. C. for one week, said weight percent based
upon total weight of amifostine.
[0055] In another embodiment, the dosage form of this invention
exhibits less than 5 weight percent degradation when maintained at
25.degree. C. for one month, said weight percent based upon total
weight of amifostine; preferably less than 4 weight percent
degradation when maintained at 25.degree. C. for one month, said
weight percent based upon total weight of amifostine; more
preferably less than 3 weight percent degradation when maintained
at 25.degree. C. for one month, said weight percent based upon
total weight of amifostine; and most preferably less than 2 weight
percent degradation when maintained at 25.degree. C. for one month,
said weight percent based upon total weight of amifostine.
[0056] In a further embodiment, the dosage form of this invention
exhibits less than 3 weight percent degradation when maintained at
5.degree. C. for about two years, said weight percent based upon
total weight of amifostine; preferably less than 2 weight percent
degradation when maintained at 5.degree. C. for about two years,
said weight percent based upon total weight of amifostine; and more
preferably less than 1 weight percent degradation when maintained
at 5.degree. C. for about two years, said weight percent based upon
total weight of amifostine.
[0057] In a further embodiment, this invention is directed to a
thermally-stable dosage form comprising substantially amorphous
amifostine. Preferably, the thermally-stable dosage form comprises
amifostine that is at least 80% amorphous by weight. More
preferably, the thermally-stable dosage form comprises amifostine
that is at least 90% amorphous by weight. This invention is also
directed to a thermally-stable dosage form comprising amorphous
amifostine.
[0058] In another embodiment, this invention is directed to a
thermally-stable, substantially amorphous dosage form comprising
amifostine. Preferably, the thermally-stable dosage form is at
least 80% amorphous by weight. More preferably, the
thermally-stable dosage form is at least 90% amorphous by weight.
This invention is also directed to an amorphous thermally-stable
dosage form comprising amifostine.
[0059] Another embodiment of this invention provides a
thermally-stable dosage form that contains an amount of water
between about 0 and about 20 weight percent, preferably between
about 0 and about 15 weight percent, and more preferably between
about 0.5 and about 6 weight percent.
[0060] The present invention is further directed to
thermally-stable dosage form of amifostine which comprises one or
more stabilizers and optionally one or more excipients. Preferably,
the molar ratio of stabilizer to amifostine is between about 0.05
and about 5.0, more preferably between about 0.1 and about 1.0, and
most preferably between about 0.2 and about 0.5 weight percent.
[0061] As above, the thermally-stable dosage form which comprises
one or more stabilizers and optionally one or more excipients may
also contain an amount of water between about 0 and about 20 weight
percent, preferably between about 0 and about 15 weight percent,
and more preferably between about 0.5 and about 6 weight percent.
Each of the dosage forms of the invention can be produced as an
elegant cake product (e.g., dissolves in less than 60 seconds).
[0062] This invention is further directed to a process of making a
thermally-stable dosage form of amorphous amifostine. In this
process, amifostine may be combined with one or more stabilizers
and/or one or more excipients. As made clear below, this method
encompasses all means by which amifostine, water, stabilizers, and
excipients may be combined. For example, amifostine and a
stabilizer may be combined in any order prior to, during, formed in
situ, or after lyophilization of bulk amifostine. Because several
different stabilizers may be used to prepare the dosage form of
this invention, each may be combined with amifostine together or
separately in any order during its preparation.
[0063] It has been found, however, that an especially economical
and efficient means of creating a thermally-stable dosage form of
amorphous amifostine (e.g., Amorphous Amifostine II) involves the
formation of a single solution comprising bulk amifostine, a
stabilizer, and optionally an excipient. According to the present
invention, vials are loaded with a sterile filtered solution
comprising amifostine and optionally a stabilizer and/or excipient.
A particular advantage of this processes is that the solvent used
to form the solution may be pure water, instead of the
ethanol/water mixture required for the production of sterile
crystalline amifostine compositions. This invention, however,
encompasses the use of other solvents, and in particular aqueous
solvents having pHs of between about 3 and about 11, more
preferably between about 6 and about 9, and most preferably between
about 7 and about 8. Bulk amifostine is known to be more stable in
basic solutions. Risley, J. M. and Van Etten, R. L., Biochem.
Pharmacol. 35:1453-1458 (1986).
[0064] After being filled with the amifostine solution, the vials
are loaded into a freeze-dryer which is then evacuated. The
freeze-drier may be pre-chilled. The vials are kept at a
temperature and pressure sufficient to remove a significant portion
of the solvent. Typically, and as is well known to those skilled in
the art, the temperature of the vials is increased near the end of
the freeze-drying cycle to ensure that the desired amount of
solvent is removed. The novel drying process described in detail
below, however, has been found to form cake products that have
consistencies and stabilities previously thought not to be possible
for non-crystalline dosage forms. Furthermore, the drying process
of this invention helps ensure that if the dosage form of this
invention (e.g., Amorphous Amifostine II) is to contain a
stabilizer, the maximum stabilizing effect of that stabilizer is
obtained.
[0065] Although the following freeze drying conditions are not
meant to be limiting, it has been found that Amorphous Amifostine
II is most easily formed when the freeze drying, or lyophilization,
process comprises the following steps: freezing the amifostine
solution during a freezing cycle to form a frozen mixture;
evacuating (vacuum) the lyophilization chamber; and drying the
frozen mixture during a primary drying cycle which comprises at
least one primary drying stage. The process may further comprise
further drying using a secondary drying cycle comprising at least
one secondary drying stage, and/or a desiccating means such as the
use of desiccants, a desiccator, desiccating stoppers, and the
like. Upon completion of the lyophilization, an inert gas such as
nitrogen is preferably added to the vials containing the
lyophilized dosage form.
[0066] It is preferred that the freezing cycle last from between
about 1 hour and about 15 hours, more preferably from between about
2 hours and about 10 hours, and most preferably between about 5
hours and about 8 hours. It is preferred that temperature of the
freeze dryer during the freezing cycle be maintained at an average
of between about -70.degree. C. and about -10.degree. C., more
preferably between about -65.degree. C. and about -30.degree. C.,
and most preferably between about -55.degree. C. and about
-40.degree. C.
[0067] Near the end, or upon completion, of the freezing cycle, the
freeze drier is evacuated such that the frozen mixtures are exposed
to a pressure (vacuum) sufficient to remove the solvent (which may
exist in a liquid and/or solid phase) at the average temperature of
the primary drying cycle. When the solvent is water, preferred
pressures are less than the vapor pressure of water at the
particular drying cycle temperature. Such vapor pressures are
provided by the CRC Handbook of Chemistry and Physics, page D-198
(64.sup.th ed; 1983-1984).
[0068] The freeze drier may instead be evacuated upon or following
commencement of the primary drying cycle. It is during this cycle
that most of the water or solvent in the frozen mixture is removed.
This cycle is characterized by an average pressure, average
temperature and duration, but may contain one or more stages, each
of which is characterized by an average pressure, average
temperature and duration. Typically, however, it is preferred that
the primary drying cycle comprise only one stage that lasts from
between about 8 hours and about 100 hours, more preferably from
between about 16 hours and about 86 hours, and most preferably
between about 24 hours and about 72 hours. It is preferred that
temperature of the freeze dryer during the primary drying cycle be
maintained at an average of between about -45.degree. C. and about
10.degree. C., more preferably between about -35.degree. C. and
about 0.degree. C., and most preferably between about -30.degree.
C. and about -10.degree. C.
[0069] The primary drying cycle is optionally followed by a
secondary drying cycle, during which residual water or solvent is
removed. This cycle is characterized by an average pressure,
average temperature and duration, but may contain one or more
stages, each of which is characterized by an average pressure,
average temperature and duration. Typically, however, it is
preferred that the secondary drying cycle comprise only one stage
that lasts from between about 0.5 hours and about 48 hours, more
preferably from between about 2 hours and about 36 hours, and most
preferably between about 6 hours and about 24 hours. It is
preferred that temperature of the freeze dryer during the secondary
drying cycle be maintained at an average of between about
-25.degree. C. and about 40.degree. C., more preferably between
about -15.degree. C. and about 40.degree. C., and most preferably
between about 0.degree. C. and about 35.degree. C.
[0070] Instead of a secondary drying cycle, or in addition thereto,
an optional desiccating means may be used to further dry the dosage
forms. Desiccating means include, but are not limited to, the use
of desiccants, desiccators, desiccating stoppers, and the like.
[0071] For example, an elegant cake product has been found to form
if vials are initially loaded into a freeze drier kept at a
temperature of between about -20.degree. C. and about 15.degree.
C., after which the freeze drier shelf temperature is decreased to
between about -70.degree. C. and about -30.degree. C. over a period
of time that allows uniform freezing of the amifostine/stabilizer
solution. The freeze drying chamber is then evacuated to cause
removal of the solvent. It is preferred that the pressure be
maintained at between about 10 .mu.m Hg to about 500 .mu.m Hg, more
preferably between about 75 .mu.m Hg and about 200 .mu.m Hg if the
solvent is water. Typically, after the pressure is allowed to
equilibrate the temperature of the chamber is increased to a
temperature of between about -35.degree. C. to about 25.degree. C.
for a time sufficient to remove most of the solvent. As described
in more detail below, it has been found that a suitable cake
product is best formed by exposing the vials to a variety of
different temperatures and pressures over periods of time ranging
from minutes to days.
[0072] Completion of the lyophilization process yields a stable
amifostine dosage form comprising amorphous amifostine that is
suitable for reconstitution into a particulate-free solution
suitable for parenteral administered to a patient. It is preferred,
however, that the freeze-drying chamber be filled with an inert gas
such as nitrogen, argon or xenon prior to the sealing of the vials.
It has been found that this facilitates the stability of the dosage
form. The vials can then be stored and shipped at a temperature
that depends upon the desired shelf life of the dosage form. A
particularly suitable temperature is between about 0.degree. C. to
about 25.degree. C., most preferably about 4.degree. C.
[0073] It has been found that by varying the temperature, time and
number of drying cycles, products with different stabilities and
water contents can be obtained, all of which are encompassed by the
present invention. Karl Fisher titration, for example, may be used
to determine water content.
[0074] According to the present invention, any amount of stabilizer
may be combined with amifostine just so long as that amount is
sufficient to increase the thermal stability of the lyophilized
product. It is preferred, however, that between about 0.05 mol to
about 5.0 mol of stabilizer be used per 1.0 mol pure bulk
crystalline amifostine trihydrate (FW=268 g/mol). This amount is
more preferably between about 0.1 mol to about 1.0 mol, and most
preferably between about 0.2 mol to about 0.5 mol bulk crystalline
amifostine trihydrate. As those skilled in the art will recognize,
however, the exact amount of stabilizer added depends upon a
variety of factors, including the chemical and pharmaceutical
characteristics of the stabilizer and the purity and water content
of the bulk amifostine. The latter is particularly noteworthy, as
bulk amifostine is available in crystalline forms, and in varying
degrees of hydration. Bulk amifostine has been described as a
crystalline form having from one to three water molecules per
molecule, but may also exist in other forms. Bulk amifostine may be
made by the process disclosed in U.S. Pat. No. 3,892,824, which is
hereby incorporated by reference.
[0075] Stabilizers of the present invention are compounds or
mixtures of compounds that are pharmaceutically acceptable and when
combined with amifostine do not facilitate its degradation in
solution yet enhance its stability when stored as a solid at room
temperature. Suitable compounds include amides and amines such as,
but are not limited to, nicotinamide, derivatives of nicotinamide,
nicotinic acid, derivatives of nicotinic acid, and a variety of
natural and synthetic amino acids and amino acid derivatives
including, but not limited to, glycine, alanine, valine, leucine,
isoleucine, phenylalanine, tryptophan, and asparagine.
[0076] Preferred stabilizers are nicotinamide, derivatives of
nicotinamide, nicotinic acid, and derivatives of nicotinic acid.
When nicotinamide is used as a stabilizer, it is preferably used in
an amifostine to nicotinamide weight ratio of between about 2:1 and
about 12:1, more preferably between about 4:1 and about 10:1. A
most preferred weight ratio is about 8:1 (amifostine to
nicotinamide).
[0077] It has been found that in order to obtain an elegant cake
upon lyophilization of a amifostine/stabilizer solution, the
concentration of bulk amifostine should be between about 10 mg/ml
to about 200 mg/ml, more preferably between about 50 mg/ml to about
150 mg/ml, and most preferably between about 75 mg/ml to about 125
mg/ml. The preferred concentration of stabilizer(s) may be
determined from the preferred amifostine-to-stabilizer molar ratios
provided above. As those skilled in the art will quickly recognize,
the solubility of the desired stabilizer(s), readily determined
from the literature, may also play a role in determining desired
concentrations.
[0078] It is important to note that when used in combination with
certain lyophilization methods, some stabilizers and concentrations
of stabilizers can yield crystalline or partially crystalline,
rather than purely amorphous, products. For example, when
nicotinamide is used as a stabilizer in a weight ratio of 4:1
(amifostine to nicotinamide) under lyophilization conditions
encompassed by this invention, a crystalline dosage form can
sometimes result. Further, a dosage form with crystalline or
partially crystalline stabilizer(s) and/or excipient(s) can also
result. The present invention is concerned, however, with amorphous
dosage forms or dosage forms containing amorphous amifostine.
[0079] As mentioned above, lyophilization of the bulk amifostine
may optionally occur in the absence of a stabilizer. If it is done
in the absence of a stabilizer, the stabilizer may be added
following lyophilization. Similarly, excipients may be added to the
dosage forms of the present invention either before, during, or
after sterilization and lyophilization of the bulk amifostine.
Among the excipients found useful for this purpose, often in
combination, are sodium chloride, citric acid, tartaric acid,
gelatin and carbohydrates such as dextrose, sucrose, sorbitol,
inositol, dextran and mannitol. Mannitol, however, is not
preferred. In addition to those mentioned herein others are known
to those skilled in the art, although it is preferred that the
excipient not be a source of free phosphate, as this has been
reported to facilitate the degradation of amifostine. See, e.g.,
Zadeii, J. M., et al., Pharm. Res. 8(10):S172 (1991).
[0080] Whatever excipient is incorporated into the present
formulations, that excipient must be sterile when added, or
sterilized during the same process that sterilizes the amifostine.
An embodiment of the present invention includes a sterilization
step. Sterilization may be affected, for example, by sterile
filtering a solution, e.g., through a 0.2 .mu.m pore size filter.
Other methods of sterilizing known to those skilled in the art may
also be employed. Suitable sterile and non-sterile excipients are
commercially available from: EM Industries, Inc., Hawthorne, N.Y.;
J. T Baker, Inc., Hayward, Calif.; Spectrum Quality Products, Inc.,
Gardena Calif.; Fisher Scientific International, Inc., Hampton
N.H.; Aldrich Chemical Co., Inc., Milwaukee Wis.; Abbott
Laboratories, Inc., North Chicago Ill.; Baxter Healthcare
Corporation, Deerfield Ill; and Amresco, Inc., Cleveland Ohio.
[0081] The dosage form of the invention may be provided in single
dose container forms by aseptically filling suitable containers
with the sterile solution to a prescribed amifostine content as
described above, or in multiple dose containers. It is intended
that these filled containers will allow rapid dissolution of the
solid composition upon reconstitution with appropriate sterile
diluents in situ, giving an appropriate sterile solution of desired
amifostine concentration for administration. As used herein, the
term "suitable containers" means a container capable of maintaining
a sterile environment, such as a vial, capable of delivering a
vacuum dried product hermetically sealed by a stopper means.
Additionally, suitable containers implies appropriateness of size,
considering the volume of solution to be held upon reconstitution
of the vacuum dried composition; and appropriateness of container
material, generally Type I glass. The stopper means employed, e.g.,
sterile rubber closures or an equivalent, should be understood to
be that which provides the aforementioned seal, but which also
allows entry for the purpose of introduction of diluent, eg.,
sterile Water for Injection, USP, Normal Saline, USP, or 5%
Dextrose in Water, USP, for the reconstitution of the desired
amifostine solution. These and other aspects of the suitability of
containers for pharmaceutical products such as those of the
invention are well known to those skilled in the practice of
pharmaceutical arts.
[0082] The sterile, thermally-stable pharmaceutical compositions
(dosage forms) of the present invention are suitable for parenteral
administration, for example, intravenous, intramuscular,
intracavitary, intrathecal, and subcutaneous injections. These
dosage forms have enhanced temperature stability over the prior
dosage forms of amorphous amifostine, have good physical
appearance, and are efficacious in the prevention and treatment of
radiation damage and chemotherapy damage.
EXAMPLES
[0083] The following examples are intended to be illustrative of
the present invention and should not be construed, in any way, to
be a limitation thereof.
Example 1
METHOD OF PRODUCING STABLE AMORPHOUS AMIFOSTINE DOSAGE FORM WITHOUT
EXCIPIENT
[0084] An aqueous solution of 100 mg/ml amifostine and 12.5 mg/ml
nicotinamide (Aldrich) is sterile filtered at 25.degree. C. through
a 0.2 .mu.m filter, and then divided into 5 ml aliquots, each of
which is transferred to 10 ml vials. Lyophilization stoppers are
placed on the vials and the samples are loaded onto the
freeze-dryer shelf maintained at 5.degree. C. The shelf temperature
is decreased to -45.degree. C. over 60 minutes, at which
temperature it is kept for about 3 hours. The freeze-dryer
condenser is then turned on and chamber is evacuated to about 100
.mu.m Hg. After the chamber vacuum has equilibrated, the shelf
temperature is ramped to -25.degree. C. over 60 minutes while the
vacuum is held constant. The shelf temperature is held at
-25.degree. C. for about 48 hours. The shelf temperature is then
ramped to -10.degree. C. over 60 minutes, and then maintained at
-10.degree. C. for 24 hours. Finally, the shelf temperature is
ramped to about 35.degree. C. over 60 minutes and then kept at that
temperature for about 24 hours. At this point, the chamber is
filled with nitrogen to a pressure of about 650 mm Hg and the vials
are mechanically stoppered. This procedure results in
thermally-stable, vacuum-dried single dose vials containing
approximately 500 mg of amifostine (anhydrous basis) and 62.5 mg
nicotinamide stabilizer as an elegant cake.
[0085] Dosage forms prepared according to this procedure were
placed on stability tests, and yielded the results provided in
Table 1 below.
Example 2
DETERMINATION OF THE CRYSTALLINITY OF STABLE AMIFOSTINE DOSAGE
FORMS
[0086] The crystallinity of the dosage forms of this invention may
be determined by powder x-ray diffraction as described, for
example, in Remington's Pharmaceutical Sciences, 18.sub.th ed. page
173; The United States Pharmacopeia, 23.sup.rd ed. (1995) pages
1843-1844.
[0087] FIG. 2 shows a typical powder x-ray diffraction spectrum of
a dosage form of amorphous amifostine prepared according to the
method of Example 1 which was measured with a Geiger-Muller
detector using nickel-filtered Cu K.alpha. radiation. This
diffraction pattern contains the broad baseline characteristic of
amorphous material. The peaks at 2.theta..apprxeq.14.8, 25.6, and
26.3 are attributed to nicotinamide and/or noise. This assignment
is clear from FIG. 3, which shows the x-ray powder diffraction
pattern of crystalline nicotinamide.
[0088] FIG. 4 shows the difference between the x-ray diffraction
patterns of crystalline amifostine prepared as described by U.S.
Pat. No. 5,591,731 and that of an amorphous amifostine dosage form
of the present invention wherein nicotinamide is used as a
stabilizer (8:1 amifostine to nicotinamide weight ratio).
Example 3
PREFERRED MANNER OF CONDUCTING STABILITY TEST OF STABLE AMORPHOUS
AMIFOSTINE DOSAGE FORMS
[0089] The stability of sterile amorphous amifostine dosage forms
obtained as described in Example 1 was tested at 5.degree. C. and
25.degree. C. Samples were stored in sealed, nitrogen-filled 10 ml
tubing vials.
[0090] Results of these tests may be correlated to stability at
other temperatures by conventional methods. See, e.g., L. Lachman,
et al., The Theory and Practice of Industrial Pharmacy pages 766-67
(1986), for a general discussion of stability prediction. From the
results of the tests conducted here, decomposition of a dosage form
kept at 25.degree. C. for one month is generally equal to about
one-half the decomposition of the same dosage form kept at
5.degree. C. for two years.
[0091] At the end of each testing period, the amorphous amifostine
in the vials was tested for water content, thiol content, and/or
amifostine content. In some cases, the water content was determined
by Karl Fischer titration. Because amifostine may undergo
hydrolysis under stress to produce 2-[(3-aminopropyl)amino] ethane
thiol ("WR-1065") and phosphoric acid, determination of the amount
of this thiol gives an indication of the stability of the
amifostine. Measurement of the amount of amifostine, however, is
not preferred, as it has been found that stabilizers such as
nicotinamide interfere with assays that determine the amount of
amifostine in a sample. Consequently, it is preferred that the
decomposition of a sample be determined by analysis of
decomposition products, and WR-1065 in particular. Analysis of
thiol content was conducted by high pressure liquid chromatography
(HPLC) using the following procedure:
1. Preparation of Standards and Samples
[0092] Weight and volumes may be adjusted provided the final
concentrations remain the same. Store solutions under refrigeration
and/or in a refrigerated autosampler immediately after preparation
shelf-life 24 hours.
1.1 Preparation of Amifostine Standard Solution
[0093] Accurately weigh approximately 30.0 mg of amifostine
standarded into a 10-ml volumetric flask. Dissolve in 5 ml of water
and dilute to volume with methanol.
1.2 Preparation of 2-[(3 -amino propy)amino] ethanethiol,
dihydrochloride (WR-1065) Standard Solution
[0094] Accurately weigh approximately 7 mg of WR-1065 standard into
a 100-ml volumetric flask. Dissolve and dilute to volume with
mobile phase.
1.3 Preparation of Amifostine (Drug Substance) for Amifostine
Content
[0095] Accurately weigh approximately 30.0 mg of amifostine into a
10-ml volumetric flask. Dissolve in 5 mL of water and dilute to
volume with methanol.
1.4 Preparation of Amifostine (Drug Substance) for Related
Substances
[0096] Accurately weigh approximately 150.0 mg of amifostine into a
10-ml volumetric flask. Dissolve and dilute to volume with
water.
1.5 Preparation of Amifostine for Injection
[0097] Dissolve contents of one drug product vial with about 9 ml
water. Quantitatively transfer sample to 50 ml volumetric flask and
dilute to volume with water. Transfer 6 ml of this solution to a
25-ml volumetric flask, add 6.5 ml of water and dilute to volume
with methanol.
1 2. System Suitability (Using Standard Solutions) Amifostine (Use
Standard Solution 1.1) % RSD of 6 Replicate Injection of Amifostine
.ltoreq.2% Tailing Factor .ltoreq.2 Theoretical Plates
.gtoreq.1,000 WR-1065 (Use Standard Solution 1.2) % RSD of 6
Injections .ltoreq.4 Tailing Factor .ltoreq.2 Theoretical Plates
.gtoreq.7,000
3. Equipment and Materials (As Stated Below or Equivalent
[0098] Equipment: HPLC System with UV Detector.
[0099] Materials: Amifostine Standard; WR-1065 Standard;
Concentrated Phosphoric Acid (H.sub.3PO.sub.4) (HPLC Grade);
Methanol (MeOH) (HPLC Grade); Purified Water (16 meg-ohm or
greater); and 1-Octanesulfonic Acid, Sodium Salt (OSA) (HPLC
Grade).
2 HPLC Chromatrographic Conditions Column Specifications: Packing
Material: Beckman Ultrasphere ODS (USP L1) Dimensions: 4.6 .times.
250 mm Particle Size: 5 mm Mobile Phase: Methanol/Aqueous
Phosphoric Acid, pH 2.5, 5 mM OSA (50/50) 1. Dissolve 0.54 g OSA in
500 ml of water, adjust with phosphoric acid to pH 2.5. 2. Dilute
to 1000 ml with methanol. 3. Filter and degas the mobile phase.
Detection: 220 nm Absorbance Flow Rate: 1.0 mL/min Injection
Volume: 10 .mu.L Column Temperature: Ambient Sample Temperature:
4.degree. C.
4. Procedure
[0100] Inject sample and standard solutions, record retention time
of the amifostine peak (approximately 4 minutes). Retention time of
the standard amifostine peak and the sample preparation peak should
agree within 10% to confirm identification of amifostine in the
sample.
5. Calculations
[0101] The calculations used to determine the degree of sample
decomposition are the same as those described in U.S. Pat. No.
5,591,731, which is hereby incorporated by reference.
Example 4
STABILITY RESULTS OF AMORPHOUS AMIFOSTINE II
[0102] Typical results obtained by testing a dosage form of this
invention according to the parameters described in Example 3 are
summarized in Table 1. This dosage form was made by the process
described in Example 1. As examination of this data makes clear,
the stability of the dosage forms of the present invention is
surprisingly independent of their moisture content.
3TABLE 1 temperature.dagger-dbl. time on stability (.degree. C.)
(days) % WR-1065 Moisture content 5 20 1.32 0.52% 5 20 1.43 0.50%
25 20 3.02 0.75% 25 20 3.07 0.86% 25 97 5.73 n.d. 25 97 5.94 n.d.
.dagger-dbl.Temperatures approximately accurate to .+-. 3.degree.
C. n.d. = not determined.
[0103] It has been observed that the stability of the dosage forms
of this invention can be varied by changing the length of the
drying steps, described in Example 1. For example, when the vials
are kept at -25.degree. C. for 12 hours, then maintained at
-10.degree. C. for 12 hours and finally kept at 35.degree. C. for 6
hours, samples are produced that show the stability results
provided in Table 2.
4TABLE 2 temperature.dagger-dbl. time on stability (.degree. C.)
(days) % WR-1065 Moisture content 25 27 3.68 n.d. 40 27 21.51 n.d.
40 27 14.76 n.d. 5 56 0.97 n.d. 5 56 1.03 n.d. 25 56 5.41 n.d. 25
56 5.28 n.d. 5 91 0.96 5.13% 5 91 1.12 4.35% 25 91 7.03 4.57% 25 91
6.73 3.6% .dagger-dbl.Temperatures approximately accurate to .+-.
3.degree. C. n.d. = not determined.
[0104] The above results clearly indicate the enhanced
thermal-stability of the amifostine dosage form produced by the
method described herein. The enhanced stability is evident from the
low weight percent of thiol formation, which indicates very little
decomposition of the amifostine to form 2-[(3-aminopropyl)amino]
ethane thiol (WR-1065).
[0105] The exceptional and unexpected thermal stability of the
dosage forms of this invention is also apparent from Table 3, which
provides stability measurements of conventional dosage forms of
amifostine (Amorphous Amifostine I), dosage forms of this invention
(Amorphous Amifostine II), and the crystalline dosage form of U.S.
Pat. No. 5,424,471 ("Crystalline Ethyol.RTM."). It is clear that
the dosage form of the present invention possesses stability
heretofore only exhibited by crystalline dosage forms.
5 TABLE 3 % 1 Month 3 Months 2 Years Dosage Form Water 40.degree.
C..dagger-dbl. 25.degree. C..dagger-dbl. 25.degree. C..dagger-dbl.
5.degree. C..dagger-dbl. Amorphous .about.6% n.d. 6.0 13.8 3.5
Amifostine I.sup.a Amorphous .about.1% 18.1 3.0.sup.f 5.8 n.d.
Amifostine II.sup.b Amorphous .about.5% n.d. 3.7 6.9 n.d.
Amifostine II.sup.c Amorphous .about.6% n.d. 5.0 n.d. n.d.
Amifostine II.sup.d Amorphous .about.6% n.d. 3.3 n.d. n.d.
Amifostine II.sup.e Crystalline Ethyol .RTM. .about.20% 0.6 n.d.
0.4 0.5 .sup.aComprising mannitol as an excipient in a 1:1 ratio.
.sup.bComprising nicotinamide as a stabilizer in a 8:1 ratio
(amifostine to nicotinamide). .sup.cComprising nicotinamide as a
stabilizer in a 8:1 ratio (amifostine to nicotinamide).
.sup.dComprising nicotinamide as a stabilizer in a 8:1 ratio
(amifostine to nicotinamide). .sup.eDosage form produced by the
methods of this invention comprising nicotinamide as a stabilizer
in a 4:1 ratio (amifostine to nicotinamide) (a mixture of
crystalline and amorphous). .sup.fDuration of 20 days.
.dagger-dbl.Temperatures approximately accurate to .+-. 3.degree.
C. n.d. = not determined.
[0106] Further, as can be seen from Table 3, variations in drying
cycle can affect thermal-stability. For example, Amorphous
Amifostine I was prepared using the prior method discussed above,
while the embodiments of Amorphous Amifostine II were prepared
using different drying cycles. Using the superscripts of Table 3,
Amorphous Amifostine II.sup.b was prepared using the drying cycle
of Example 1, and Amorphous Amifostine II.sup.c was prepared using
the drying cycle described above that provided the results in Table
II. Amorphous Amifostine II.sup.d and Amorphous Amifostine II.sup.e
were both prepared using the following drying cycle: freezing for
about 3 hours at a temperature of about -45.degree. C.; primary
drying for about 60 hours at a temperature of about -25.degree. C.;
and secondary drying for about 24 hours at about -10.degree. C.,
and then for about 6 hours at about 0.degree. C.
[0107] It should be apparent to one of ordinary skill that other
embodiments not specifically disclosed nonetheless fall within the
scope and spirit of the present invention. Hence, the descriptions
herein should not be taken as limiting the invention in any way,
except as stated in the following claims.
* * * * *