FIELD OFTHE INVENTION
The present invention relates to stable, ready to us;-;'on aqueous Pharmaceutical compositions
comprising 5-aza-2'deoxycitidine and at least one aprotic solvent. The pharmaceutical compositions
may further comprise at least one protic solvent. The pharmaceutical compositions thus prepared
can be used for the treatment of patients suffering from myelodysplastic syndromes.
BACKGROUND OF THE INVENTION
5-aza-2'deoxycitidine (Decitabine) is an analogue of the natural nucleoside 2'-deoxycytidine and
shown in Figure 1.
HO
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Figure 1: Structure of Decitabine
Decitabine is believed to exert its antineoplastic effects after phosphorylation and direct
incorporation into DNA and inhibition of DNA methyltransferase, causing hypomethylation of DNA
and cellular differentiation or apoptosis. Decitabine inhibits DNA methylation in vitro, which is
achieved at concentrations that do not cause major suppression of DNA synthesis. Decitabineinduced
hypomethylation in neoplastic cells may restore normal function to genes that are critical for
the control of cellular differentiation and proliferation. In rapidly diViding cells, the cyt'f'~'"> I
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Another object pf tM"present invention is to provide pharmaceutical compositions that ar~~~ ~,t~b'tt>\J'
under both real time and accelerated storage conditions. '1 ~ u'..., ~
Yet another object of the present invention is to avoid the micronization of Decitabine.
Yet another object of the invention is to provide more cost efficient and economical formulations of
decitabine.
SUMMARY OF THE INVENTION
The present invention relates to ready to use, non aqueous pharmaceutical compositions comprising
the known compound 5-aza-2'deoxycitidine and at least one aprotic solvent. The pharmaceutical
compositions may further comprise at least one protic solvent.
In one embodiment of the present invention the oxygen content is controlled by the addition of
antioxidants or by using an inert gas such as nitrogen.
In another embodiment of the present invention, Decitabine is used in unmicronized form ..
In yet another embodiment of the present invention the pharmaceutical compositions are stable
under both real time and accelerated storage conditions.
In yet another embodiment of the invention the compositions are cost efficient and economical.
DETAILED DESCRIPTION OF THE INVENTION
In one aspect the present invention relates to ready to use, non aqueous pharmaceutical
compositions comprising 5-aza-2'deoxycitidine and at least one aprotic solvent. The pharmaceutical
compositions may further comprise at least one protic solvent.
In another embodiment of the present invention the oxygen content is controlled by using
antioxidants or an inert gas such as nitrogen.
In a preferred embodiment the formulations are presented as a single vial presentation comprising
5-aza-2'deoxycitidine in a concentration of 5 mg/ml.
The pharmaceutical compositions of the present invention are suitable for parenteral administration.
These pharmaceutical compositions are then administered via intravenous infusion to treat patients
suffering from• myelodysplastic syndromes (MDS) inclUding previously treated and untreated, de
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novo and secon~ary MDS of all French-American-British subtypes (refractory anemia", refractory
anemia with ringed sideroblasts, refractory anemia with excess blasts, refractory anemia with excess
blasts in transformation, and chronic myelomonocytic leukemia) and intermediate-1, intermediate2,
and high-risk International Prognostic Scoring System groups.
In one embodiment of the present invention, the aprotic solvent is selected from the group consisting
of 1-methyl-2-pyrrolidone, 1,3-dimethyl-2-imidazolidinone, dimethylacetamide, dimethyl sulfoxide,
acetone, tetrahydrofuran, 1,4-dioxane, acetonitrile, dimethyl formamide, propylene carbonate, or
mixtures thereof. Dimethylacetamide, dimethyl sulfoxide, or mixtures thereof are preferred.
Dimethylacetamide is especially preferred.
In another embodiment of the present invention, the protic solvent is selected from the group
consisting of alcohols, amides, or mixtures thereof. Alcohols, for example ethanol, are• especially
preferred.
In yet another embodiment of the present invention, the oxygen content is controlled by using
antioxidants or inert gas such as nitrogen. This may be aided by, for example, purging the sealable
container with a gas which is substantially oxygen-free, or substantially moisture free, or
substantially oxygen and moisture free. Purging can be expected to reduce the oxygen level in the
sealable container to a level of from about 0.5% to about 1DOlo, typically about 5% or lower,
depending on the efficiency of flushing and how qUickly the container is sealed after flushing. The
gas used for purging the sealable container may be any appropriate inert gas known to those in the
art, the most commonly used gases being argon, helium or nitrogen, or mixtures thereof. However
the most preferred inert gas is nitrogen.
In yet another embodiment of the present invention decitabine is not micronized. It has surprisingly
been found that the micronization step is not necessary when preparing the compositions according
to the present invention. Hence, these are more cost efficient and economical.
The invention is further illustrated by the following examples, which are not construed to be
limiting the scope of the invention.
5
Composition according to example 1 containing dimethylacetamide
and ethanol in the ratio of 70:30
Table 1:
EXAMPLES
,~_.': ,..~~"r1(" Example 1 . " ~;; .; _'
The composition of the present invention contains decitrbi~fu a conc~~t;ation of 5 mg/mL in
a solvent system comprising dimethylaceta"mide and ethanol in the ratio of 70:30. The details
of the composition are shown inTable 1.
Sr. No. Ingredients Quantity
(mg/mL)
1 Decitabine 5 mg
2 Dimethylacetamide 0.7mL
3 Ethanol qs to 1mL
.. - 4 Nitrogen* qs
* Nitrogen is used for purging in bulk solution and blanketing in vial headspace
The pharmaceutical composition according to example 1 was prepared by the following
process:
A suitable quantity of dimethylacetamide was Jed into a manufacturing vessel and
subsequently the required quantity of alcohol was added. The two components were then
mixed. Nitrogen was then purged into the solution obtained in the previous step until the
oxygen content was below 7 mg/L, preferably below 3 mg/L. Decitabine was added and
stirred in about 80 % of the solution obtained in the previous step and dissolved. The
volume was then made up to 100% with the solution obtained in step 2. The resulting
Decitabine solution was then filtered through a suitable sterilizing grade filter and filled into
vials. The"vial headspace was then blanketed with nitrogen to achieve a headspace
oxygen content of less than 10 %, preferably less than 5 %, more preferably less than 2 %.
Finally the vials were stoppered and sealed.
The stability profile of the formulation according example 1 was analysed and is presented
in Table 2. The amount of Decitabine in the compostion was measured before and after
storage. The term "assay" as used in table 2 refers to the quantitative determination of
decitabine via HPLC. Also, the impurity profile of the composition was analyzed before and
after storage at various temperature and humidity conditions.
6
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Table 2: Stability data of the composition according to example 1
Related Substances (% w/w)
Assay (%) Water by KF Total impurity
Stability Conditions .
Initial 102.8 0.42 0.15
40oC/75%RH 100.9 0.34 0.44
(7 days)
Accelerated stability
condition 102.9 0.24 0.85
250C/60%RH (3M)
Accelerated stability
condition 102.1 0.26 0.90
250C/60%RH (6M)
Real time stability
condition 103.4 0.23 0.36
2-8°C (3M)
Real time stability
condition 102.9 0.28 0.27
2-8°C (6M)
Example 2
The composition of the present inven•tion contains Decitabine in a concentration of 5
mg/mL in a solvent system comprising dimethylacetamide and ethanol in the ratio of 60:40
. The details of the composition are given in table 3.
Table 3: Composition according to example 2 containing dimethylacetamide and ethanol in
the ratio of 60:40
Sr. No. Ingredients Quantity
(mg/mL)
1 Decitabine 5mg
2 Dimethylacetamide 0.6mL
3 Ethanol qs to 1mL
4 Nitrogen* qs
o• * Nitrogen IS used for purging In bulk solution and blanketIng In vial headspace
The pharmaceutical composition according to example 2 was prepared by the following
process:
7
The stability profile of the formulation according to example 2 was analyzed by subjecting
the samples to various conditions and is presented in table 4. The amount of Decitabine in
the compostiori was measured before and after storage. (The term "assay" as used in table
2 refers to the quantitative determination of decitabine via HPLC.)Also, the impurity profile
of the formulation was analyzed before and after storage at various conditions.
Table 4: Stability Data of Representative Formulation (Example-2)
Stability Conditions Assay (Ofo) Water by KF
Related Substances (% w/w)
Total impurity
Initial 100.0 0.41 0.13
40uC/750f0RH
100.2 0.43 0.49
(7 days)
40uC1750f0RH
102.9 0.31 1.09
(1 month)
Accelerated stability
condition 100.4 0.46 0.83
250C/60%RH (3M)
Accelerated stability
condition 98.9 0.48 1.08
250C/60%RH (6M)
Real time stability
condition 101.1 0.47 0.28
2-8°C (3M)
Real time stability
condition 100.3 0.47 0.20
2-8°C (6M)
8
ORIGIf\"From the stability data provided in the tables above it is apparent that the pharmaceutical
compositions according to the present invention are stable under various storage
conditions (both real time and accelerated stability conditions).
Also, the impurity profile is better than that of the reference formulation, the commercially
available lyophilized product Dacogen®, under accelerated storage conditions.

We claim:
1. A ready to use, non aqueous pharmaceutic:al ~t~Position 'com~~iSing 5-aza2'
deoxycitidine and at least one aprotic solvent, wherein the aprotic solvent is selected
from the group consisting of 1-methyl-2-pyrrolidone, 1,3-dimethyl-2-imidazolidinon
dimethylacetamide, dimethyl sulfoxide, acetone, tetrahydrofuran, 1,4-dioxane,
acetonitrile, dimethyl formamide, propylene carbonate, or a mixture thereof.
2. The composition according to claim 1, wherein the aprotic solvent is dimethylacetamide.
3. The composition according to claim 1 further comprising at least one protic solvent.
4. The composition according to claim 3, wherein the protic solvent is selected from the
group consisting of alcohols, amides, or mixtures thereof.
5. The composition according to claim 4, wherein the protic solvent is ethanol.
6. The composition according to any of the preceding claims, wherein the protic and aprotic
solvents are present in a ratio of 10:90 to 50:50. preferably in a ratio of 30: 70 to 40:60.
7. The composition according to any of the preceding claims, wherei!),5-aza- 2'deoxycitidine
is present in a concentration of 5 mg/ml.
8. A method of preparing the composition according to claim 1 comprising the steps of:
a) dissolving a protic solvent in a suitable quantity of an aprotic solvent
b) purging nitrogen into the solution obtained in the previous step until the oxygen
content is below 7 mg/L, preferably below 3 mg/L
c) dissolving the drug (5-aza- 2'deoxycitidine) in about 80 % of the solution obtained
in step b)
d) making up the volume up to 100% with solution obtained in step b)
e) filtering the drug solution and filling it into vials
f) blanketing the vial headspace with nitrogen to achieve a headspace oxygen content of
less than 10 %, preferably less than 5 %, and more preferably Jess than 2 %.
g) stoppering and sealing the vials