U.S. patent application number 15/222157 was filed with the patent office on 2017-02-02 for pharmaceutical composition with improved storage stability and method for preparing the same.
This patent application is currently assigned to SAMYANG BIOPHARMACEUTICALS CORPORATION. The applicant listed for this patent is SAMYANG BIOPHARMACEUTICALS CORPORATION. Invention is credited to Joong Woong CHO, In Ja CHOI, Bong Oh KIM, Hye Rim KIM, Ji Yeong KIM, Sa Won LEE, Bum Chan MIN, Min Hyo SEO, Yil Woong YI.
Application Number | 20170028068 15/222157 |
Document ID | / |
Family ID | 57885791 |
Filed Date | 2017-02-02 |
United States Patent
Application |
20170028068 |
Kind Code |
A1 |
KIM; Bong Oh ; et
al. |
February 2, 2017 |
PHARMACEUTICAL COMPOSITION WITH IMPROVED STORAGE STABILITY AND
METHOD FOR PREPARING THE SAME
Abstract
A pharmaceutical composition containing a specific related
compound within a specified limit and a method for preparing the
same are provided.
Inventors: |
KIM; Bong Oh; (Daejeon,
KR) ; MIN; Bum Chan; (Daejeon, KR) ; KIM; Ji
Yeong; (Hanam-si, KR) ; KIM; Hye Rim;
(Daejeon, KR) ; SEO; Min Hyo; (Daejeon, KR)
; LEE; Sa Won; (Daejeon, KR) ; YI; Yil Woong;
(Daejeon, KR) ; CHO; Joong Woong; (Daejeon,
KR) ; CHOI; In Ja; (Daejeon, KR) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SAMYANG BIOPHARMACEUTICALS CORPORATION |
Seoul |
|
KR |
|
|
Assignee: |
SAMYANG BIOPHARMACEUTICALS
CORPORATION
Seoul
KR
|
Family ID: |
57885791 |
Appl. No.: |
15/222157 |
Filed: |
July 28, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62198457 |
Jul 29, 2015 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 47/32 20130101;
A61K 47/34 20130101; A61K 9/1075 20130101; A61K 31/337 20130101;
A61K 9/0019 20130101 |
International
Class: |
A61K 47/34 20060101
A61K047/34; A61K 31/337 20060101 A61K031/337; A61K 47/32 20060101
A61K047/32; A61K 9/107 20060101 A61K009/107 |
Claims
1. A polymeric micelle pharmaceutical composition, comprising: a
purified amphiphilic block copolymer comprising a hydrophilic block
(A) and a hydrophobic block (B), and one or more poorly
water-soluble drugs selected from the group consisting of
paclitaxel and docetaxel, wherein the pharmaceutical composition
contains, when stored at 40.degree. C. for 6 months, a related
compound represented by the following Formula 1 in an amount of
less than 0.58 part by weight, based on 100 parts by weight of the
initial amount of the poorly water-soluble drug: ##STR00007##
wherein R.sub.1 is H or COCH.sub.3, and R.sub.2 is phenyl or
O(CH.sub.3).sub.3.
2. The pharmaceutical composition according to claim 1, wherein the
compound of Formula 1 includes a compound of the following Formula
1a, a compound of the following Formula 1b, or both of them:
##STR00008## wherein R.sub.1 and R.sub.2 are the same as defined in
claim 1.
3. The pharmaceutical composition according to claim 2, which
contains the compound of Formula 1a in an amount of less than 0.22
part by weight, based on 100 parts by weight of the initial amount
of the poorly water-soluble drug.
4. The pharmaceutical composition according to claim 2, which
contains the compound of Formula 1b in an amount of less than 0.36
part by weight, based on 100 parts by weight of the initial amount
of the poorly water-soluble drug.
5. The pharmaceutical composition according to claim 1, wherein the
compound of Formula 1 is the compound of the following Formula 1c:
##STR00009##
6. The pharmaceutical composition according to claim 5, wherein the
compound of Formula 1c includes the compound of the following
Formula 1d, the compound of the following Formula 1e, or both of
them: ##STR00010##
7. The pharmaceutical composition according to claim 1, which
contains the related compound of Formula 1 in an amount of 0.5 part
by weight or less, based on 100 parts by weight of the initial
amount of the poorly water-soluble drug.
8. The pharmaceutical composition according to claim 7, which
contains the related compound of Formula 1 in an amount of 0.2 part
by weight or less, based on 100 parts by weight of the initial
amount of the poorly water-soluble drug.
9. The pharmaceutical composition according to claim 8, which
contains the related compound of Formula 1 in an amount of 0.1 part
by weight or less, based on 100 parts by weight of the initial
amount of the poorly water-soluble drug.
10. The pharmaceutical composition according to claim 1, which
contains, when stored at 80.degree. C. for 3 weeks, the related
compound of Formula 1 in an amount of less than 0.45 part by
weight, based on 100 parts by weight of the initial amount of the
poorly water-soluble drug.
11. The pharmaceutical composition according to claim 1, wherein
the hydrophilic block (A) comprises one or more selected from the
group consisting of polyethylene glycol or derivatives thereof,
polyvinylpyrrolidone, polyvinyl alcohol, polyacrylamide and
combinations thereof.
12. The pharmaceutical composition according to claim 1, wherein
the hydrophobic block (B) comprises one or more selected from the
group consisting of polylactide, polyglycolide, polymandelic acid,
polycaprolactone, polydioxan-2-one, polyamino acid, polyorthoester,
polyanhydride, polycarbonate and combinations thereof.
13. The pharmaceutical composition according to claim 1, wherein
the hydrophilic block (A) is polyethylene glycol or
monomethoxypolyethylene glycol, and the hydrophobic block (B) is
polylactide.
14. The pharmaceutical composition according to claim 1, wherein
the hydrophilic block (A) has a number average molecular weight of
200 to 20,000 Daltons, and the hydrophobic block (B) has a number
average molecular weight of 200 to 20,000 Daltons.
15. The pharmaceutical composition according to claim 1, wherein
the amphiphilic block copolymer is one purified by sublimation at a
temperature of 80.degree. C. or higher and lower than 120.degree.
C. and under a pressure of a vacuum degree of 10 torr or less for
10 to 74 hours.
Description
TECHNICAL FIELD
[0001] The present disclosure relates to a pharmaceutical
composition with improved storage stability and a method for
preparing the same, and more specifically, a pharmaceutical
composition of poorly water-soluble drug comprising an amphiphilic
block copolymer wherein the content of a specific related compound
is kept within a specified limit, and a method for preparing the
same.
BACKGROUND ART
[0002] Solubilization of a poorly water-soluble drug is a key
technology for delivering the drug into the body via oral or
parenteral administration. Such solubilization methods include a
method of adding a surfactant to an aqueous solution to form
micelles and then entrapping a poorly water-soluble drug therein.
An amphiphilic block copolymer used as a surfactant comprises a
hydrophilic polymer block and a hydrophobic polymer block. Since
the hydrophilic polymer block directly contacts blood proteins and
cell membranes in vivo, polyethylene glycol or
monomethoxypolyethylene glycol, etc. having biocompatibility has
been used. The hydrophobic polymer block improves affinity to a
hydrophobic drug, and polylactide, polyglycolide,
poly(lactic-glycolide), polycaprolactone, polyamino acid or
polyorthoester, etc. having biodegradability has been used. In
particular, polylactide derivatives have been applied to drug
carriers in various forms because they have excellent
biocompatibility and are hydrolyzed into harmless lactic acid in
vivo. Polylactide derivatives have various physical properties
depending on their molecular weights, and have been developed in
various forms such as microsphere, nanoparticle, polymeric gel and
implant agent.
[0003] U.S. Pat. No. 6,322,805 discloses a composition for
delivering a poorly water-soluble drug consisting of a polymeric
micelle-type drug carrier and a poorly water-soluble drug, wherein
the polymeric micelle-type drug carrier is formed from a diblock or
triblock copolymer which is not crosslinked by a crosslinking agent
and consists of at least one biodegradable hydrophobic polymer
selected from the group consisting of polylactide, polyglycolide,
poly(lactide-glycolide), polycaprolactone and derivatives thereof
and poly(alkylene oxide) as a hydrophilic polymer, wherein the
poorly water-soluble drug is physically entrapped in the drug
carrier and solubilized, and wherein the polymeric micelle-type
drug carrier forms a clear aqueous solution in water and
effectively delivers the poorly water-soluble drug into the body.
According to the above US patent, polyethylene glycol-polylactide
diblock copolymer is synthesized by removing moisture from
monomethoxypolyethylene glycol, adding stannous octoate dissolved
in toluene thereto and removing toluene under reduced pressure,
adding D,L-lactide to the resulting mixture and conducting a
polymerization reaction, adding chloroform to dissolve the produced
block copolymer, dropwise adding an excess amount of diethyl ether
in small portions with stirring to form precipitant and filtering
the formed precipitant, and washing it several times with diethyl
ether. However, this method is difficult to employ in mass-scale
production and thus is not commercially available. In addition, the
ether that has been used for purification may remain in the final
polymeric micelle composition.
[0004] U.S. Pat. No. 8,853,351 discloses a method for preparing an
amphiphilic block copolymer, comprising (a) dissolving the
amphiphilic block copolymer in a water-miscible organic solvent;
(b) adding and mixing an aqueous solution of alkali metal salt
(sodium bicarbonate, sodium carbonate, potassium bicarbonate,
potassium carbonate or lithium carbonate) to the polymeric solution
obtained in step (a); (c) separating organic and aqueous phases by
salting out for the solution obtained in step (b); and, (d)
isolating the organic phase obtained in step (c) and removing the
organic solvent therefrom to recover the polymer. However, the
method involves complicated steps, and requires an additional step
for removing the alkali metal salt and the salt (sodium chloride or
potassium chloride) used for salting out, and may have residual
metal salts even after the removal thereof.
[0005] Impurities of drug must be strictly controlled in various
aspects. Particularly, in case of impurities derived from active
pharmaceutical ingredient (API), each country determines in its
drug approval guideline the upper limit to amount of API-derived,
known or unknown impurities (related compounds) in a drug product.
In addition, there are some standards used internationally and ICH
guideline Q3A is the representative one. In this guideline, at the
time of approving a drug, the amount of each related compound in
the drug is limited up to 0.1% or 0.2%, etc. and information such
as toxicity-related data, etc., which should be provided, is
discriminately applied according to the related compound exceeding
the limit. This implies that since it is unknown how a related
compound of a drug would act in vivo, the amount of the related
compound must be reduced in the procedure of manufacturing the
drug. Therefore, a manufacturing process for reducing the related
compounds and setting of the upper limit to amount according to the
characteristics (structure and toxicity) of each related compound
are essential factors in quality control of the drug.
CONTENTS OF THE INVENTION
Problems to be Solved
[0006] One purpose of the present invention is to provide a
polymeric micelle-type pharmaceutical composition of poorly
water-soluble drug comprising an amphiphilic block copolymer, which
contains a specific related compound in an amount within a
specified limit.
[0007] The other purpose of the present invention is to provide a
method for preparing said pharmaceutical composition.
Technical Means to Solve the Problems
[0008] One aspect of the present invention provides a polymeric
micelle pharmaceutical composition, comprising: a purified
amphiphilic block copolymer comprising a hydrophilic block (A) and
a hydrophobic block (B), and one or more poorly water-soluble drugs
selected from the group consisting of paclitaxel and docetaxel,
wherein the pharmaceutical composition contains, when stored at
40.degree. C. for 6 months, a related compound represented by the
following Formula 1 in an amount of less than 0.58 part by weight,
based on 100 parts by weight of the initial amount of the poorly
water-soluble drug:
##STR00001##
[0009] wherein
[0010] R.sub.1 is H or COCH.sub.3, and R.sub.2 is phenyl or
O(CH.sub.3).sub.3.
[0011] Another aspect of the present invention provides a method
for preparing a polymeric micelle pharmaceutical composition,
comprising: (a) purifying an amphiphilic block copolymer comprising
a hydrophilic block (A) and a hydrophobic block (B); (b) dissolving
one or more poorly water-soluble drugs selected from the group
consisting of paclitaxel and docetaxel, and the purified
amphiphilic block copolymer in an organic solvent; and (c) adding
an aqueous solvent to the solution obtained in step (b) to form
polymeric micelles; wherein the pharmaceutical composition
contains, when stored at 40.degree. C. for 6 months, a related
compound represented by the above Formula 1 in an amount of less
than 0.58 part by weight, based on 100 parts by weight of the
initial amount of the poorly water-soluble drug.
EFFECTS OF THE INVENTION
[0012] According to the present invention, a pharmaceutical
composition of poorly water-soluble drug, which has reduced related
compounds and improved storage stability, can be obtained.
BRIEF EXPLANATION OF THE DRAWINGS
[0013] FIG. 1 is the resulting chromatogram of HPLC analysis for
the polymeric micelle composition containing paclitaxel used in
Experimental Example 1-1, which had been subjected to the six-month
acceleration test.
[0014] FIG. 2 is the resulting chromatogram and spectrum of
LC/MS/MS analysis conducted in Experimental Example 1-2 for the
related compound isolated in Experimental Example 1-1:
[0015] (a) RRT 1.10.+-.0.02 (1.08-1.12) (hereinafter, RRT 1.10 is
used interchangeably with RRT 1.10.+-.0.02)
[0016] (b) RRT 1.12.+-.0.02 (1.10-1.14) (hereinafter, RRT 1.12 is
used interchangeably with RRT 1.12.+-.0.02)
[0017] FIG. 3 is the resulting chromatogram of HPLC analysis for
the induced reaction product obtained in Experimental Example
2:
[0018] (a) Polymeric micelle pharmaceutical composition containing
paclitaxel
[0019] (b) Paclitaxel
[0020] (c) Reaction product of paclitaxel and L-lactide
[0021] (d) Reaction product of paclitaxel and D-lactide
[0022] FIG. 4 is the resulting chromatogram of LC/MS/MS analysis
for the induced reaction product obtained in Experimental Example
3:
[0023] (a) Paclitaxel
[0024] (b) Reaction product of paclitaxel and L-lactide
[0025] (c) Reaction product of paclitaxel and D-lactide
[0026] FIG. 5 is the resulting chromatogram of HPLC analysis
conducted in Experimental Example 4.
DETAILED DESCRIPTION TO CARRY OUT THE INVENTION
[0027] The present invention is explained in more detail below.
[0028] The pharmaceutical composition of an embodiment of the
present invention comprises a purified amphiphilic block copolymer
comprising a hydrophilic block (A) and a hydrophobic block (B).
[0029] According to one embodiment of the present invention, the
amphiphilic block copolymer comprises an A-B type diblock copolymer
consisting of a hydrophilic block (A) and a hydrophobic block (B),
or a B-A-B type triblock copolymer.
[0030] According to one embodiment of the present invention, the
amphiphilic block copolymer may comprise the hydrophilic block in
an amount of 20 to 95% by weight, and more concretely 40 to 95% by
weight, based on the total weight of the copolymer. In addition,
the amphiphilic block copolymer may comprise the hydrophobic block
in an amount of 5 to 80% by weight, and more concretely 5 to 60% by
weight, based on the total weight of the copolymer.
[0031] According to one embodiment of the present invention, the
amphiphilic block copolymer may have a number average molecular
weight of 1,000 to 50,000 Daltons, and more concretely 1,500 to
20,000 Daltons.
[0032] According to one embodiment of the present invention, the
hydrophilic block is a polymer having biocompatibility and may
comprise one or more selected from the group consisting of
polyethylene glycol or derivatives thereof, polyvinylpyrrolidone,
polyvinyl alcohol, polyacrylamide and combinations thereof, and
more concretely, it may comprise one or more selected from the
group consisting of polyethylene glycol, monomethoxypolyethylene
glycol and combinations thereof. The hydrophilic block may have a
number average molecular weight of 200 to 20,000 Daltons, and more
concretely 200 to 10,000 Daltons.
[0033] According to one embodiment of the present invention, the
hydrophobic block is a polymer having biodegradability and may be a
polymer of monomers derived from alpha (.alpha.)-hydroxy acid.
Concretely, it may comprise one or more selected from the group
consisting of polylactide, polyglycolide, polymandelic acid,
polycaprolactone, polydioxan-2-one, polyamino acid, polyorthoester,
polyanhydride, polycarbonate and combinations thereof, and more
concretely, it may comprise one or more selected from the group
consisting of polylactide, polyglycolide, polycaprolactone,
polydioxan-2-one and combinations thereof. The hydrophobic block
may have a number average molecular weight of 200 to 20,000
Daltons, and more concretely 200 to 10,000 Daltons.
[0034] According to one embodiment of the present invention, an
amphiphilic block copolymer comprising a hydrophobic polymer block
of poly(alpha (.alpha.)-hydroxy acid) may be synthesized by a known
ring-opening polymerization method using a hydrophilic polymer
having hydroxyl group as an initiator, and a lactone monomer of
alpha (.alpha.)-hydroxy acid. For example, L-lactide or D,L-lactide
may be polymerized with hydrophilic polyethylene glycol or
monomethoxypolyethylene glycol having hydroxyl group as an
initiator by ring-opening. Synthesis of diblock or triblock
copolymer is possible according to the number of hydroxyl group
existing in the hydrophilic block which is the initiator. In the
ring-opening polymerization, an organometallic catalyst such as tin
oxide, lead oxide, tin octoate, antimony octoate, etc. may be used,
and tin octoate having biocompatibility is preferably used in
preparing polymer for medical use.
[0035] In an embodiment of the present invention, as the
amphiphilic block copolymer, a purified one is used. According to a
preferable embodiment of the present invention, the amphiphilic
block copolymer is one that has been purified by sublimation.
[0036] The purification by sublimation may be conducted at a
temperature of preferably 80.degree. C. or higher and lower than
120.degree. C. and more preferably 80 to 100.degree. C., and under
a pressure of a vacuum degree of preferably 10 torr or less, more
preferably 5 torr or less and even more preferably 1 torr or less,
for a time of preferably 10 to 74 hours, more preferably 10 to 48
hours and even more preferably 24 to 48 hours. Conducting the
purification by sublimation under such conditions can minimize the
change in molecular weight of the copolymer and remove impurities
therefrom.
[0037] The pharmaceutical composition of an embodiment of the
present invention comprises, as active ingredient, one or more
poorly water-soluble drugs selected from the group consisting of
paclitaxel and docetaxel.
[0038] According to one embodiment of the present invention, the
pharmaceutical composition may further comprise, as additional
active ingredient, one or more poorly water-soluble drugs other
than paclitaxel and docetaxel. As such an additional active
ingredient, one or more taxane anticancer agents selected from the
group consisting of 7-epipaclitaxel, t-acetylpaclitaxel,
10-desacetylpaclitaxel, 10-desacetyl-7-epipaclitaxel,
7-xylosylpaclitaxel, 10-desacetyl-7-glutarylpaclitaxel,
7-N,N-dimethylglycylpaclitaxel, 7-L-alanylpaclitaxel and
cabazitaxel, may be used.
[0039] The pharmaceutical composition of an embodiment of the
present invention may comprise the poorly water-soluble drug in an
amount of 0.1 to 50 parts by weight, and more concretely 0.5 to 30
parts by weight, based on 100 parts by weight of the amphiphilic
block copolymer. If the amount of the poorly water-soluble drug is
too small as compared with that of the amphiphilic block copolymer,
the weight ratio of the amphiphilic copolymer used per drug is high
and thus the time for reconstitution may increase. On the other
hand, if the amount of the poorly water-soluble drug is too large,
there may be a problem of rapid precipitation of the poorly
water-soluble drug.
[0040] As used herein, the "initial" amount of the poorly
water-soluble drug means the weight of the poorly water-soluble
drug incorporated when the pharmaceutical composition was
prepared.
[0041] In an embodiment of the present invention, the
pharmaceutical composition contains, when stored at the accelerated
condition (40.degree. C.) for 6 months, a related compound
represented by the following Formula I in an amount of less than
0.58 part by weight, based on 100 parts by weight of the initial
amount of the poorly water-soluble drug:
##STR00002##
[0042] wherein
[0043] R.sub.1 is H or COCH.sub.3, and R.sub.2 is phenyl or
O(CH.sub.3).sub.3.
[0044] The compound of Formula 1 may include a compound of the
following Formula 1a, a compound of the following Formula 1b, or
both of them:
##STR00003##
[0045] In the above Formulas 1a and 1b, R.sub.1 and R.sub.2 are the
same as defined in Formula I above.
[0046] According to one embodiment of the present invention, the
poorly water-soluble drug is paclitaxel, and the related
compound(s) may include the compound represented by the following
Formula 1e:
##STR00004##
[0047] The compound of Formula 1c may include the compound of the
following Formula 1d, the compound of the following Formula 1e, or
both of them:
##STR00005##
[0048] The pharmaceutical composition of an embodiment of the
present invention may contain, when stored at the accelerated
condition (40.degree. C.) for 6 months, a related compound of
Formula I (particularly, Formula 1c) in an amount of less than 0.58
part by weight, for example, 0.5 part by weight or less, preferably
0.35 part by weight or less, more preferably 0.2 part by weight or
less, even more preferably 0.1 part by weight or less, and most
preferably 0.07 part by weight or less, based on 100 parts by
weight of the initial amount of the poorly water-soluble drug.
[0049] The pharmaceutical composition of an embodiment of the
present invention may contain, when stored at the accelerated
condition (40.degree. C.) for 6 months, a related compound of
Formula 1a (particularly, Formula 1d) in an amount of less than
0.22 part by weight, for example, 0.2 part by weight or less,
preferably 0.15 part by weight or less, more preferably 0.1 part by
weight or less, even more preferably 0.06 part by weight or less,
and most preferably 0.05 part by weight or less, based on 100 parts
by weight of the initial amount of the poorly water-soluble
drug.
[0050] The pharmaceutical composition of an embodiment of the
present invention may contain, when stored at the accelerated
condition (40.degree. C.) for 6 months, a related compound of
Formula 1b (particularly, Formula 1e) in an amount of less than
0.36 part by weight, for example, 0.3 part by weight or less,
preferably 0.2 part by weight or less, more preferably 0.1 part by
weight or less, even more preferably 0.04 part by weight or less,
and most preferably 0.02 part by weight or less, based on 100 parts
by weight of the initial amount of the poorly water-soluble
drug.
[0051] The pharmaceutical composition of an embodiment of the
present invention may contain, when stored at the severe condition
(80.degree. C.) for 3 weeks, a related compound of Formula 1
(particularly, Formula 1c) in an amount of less than 0.45 part by
weight, preferably 0.4 part by weight or less, more preferably 0.2
part by weight or less, and most preferably 0.16 part by weight or
less, based on 100 parts by weight of the initial amount of the
poorly water-soluble drug.
[0052] The pharmaceutical composition of an embodiment of the
present invention may contain, when stored at the severe condition
(80.degree. C.) for 3 weeks, a related compound of Formula 1a
(particularly, Formula 1d) in an amount of less than 0.18 part by
weight, preferably 0.15 part by weight or less, more preferably 0.1
part by weight or less, and most preferably 0.08 part by weight or
less, based on 100 parts by weight of the initial amount of the
poorly water-soluble drug.
[0053] The pharmaceutical composition of an embodiment of the
present invention may contain, when stored at the severe condition
(80.degree. C.) for 3 weeks, a related compound of Formula 1b
(particularly, Formula 1e) in an amount of less than 0.27 part by
weight, preferably 0.2 part by weight or less, more preferably 0.1
part by weight or less, and most preferably 0.08 part by weight or
less, based on 100 parts by weight of the initial amount of the
poorly water-soluble drug.
[0054] In an embodiment of the present invention, the
pharmaceutical composition, which contains a specific related
compound in an amount within a specified limit, is a commercially
available composition since it can be produced on a large
scale.
[0055] In an embodiment, the pharmaceutical composition of the
present invention does not have ether, for example, diethyl ether,
at all.
[0056] In an embodiment, the pharmaceutical composition of the
present invention does not have metal salt, for example, alkali
metal salt and/or salt for salting out, for example, NaCl or KCl,
at all.
[0057] The pharmaceutical composition of an embodiment of the
present invention can be prepared by a method comprising (a)
purifying an amphiphilic block copolymer comprising a hydrophilic
block (A) and a hydrophobic block (B); (b) dissolving one or more
poorly water-soluble drugs selected from the group consisting of
paclitaxel and docetaxel, and the purified amphiphilic block
copolymer in an organic solvent; and (c) adding an aqueous solvent
to the solution obtained in step (b) to form polymeric
micelles.
[0058] The purification of the amphiphilic block copolymer is
explained above, and a conventional method can be used for the
formation of the polymeric micelles.
[0059] In the method for preparing a pharmaceutical composition of
an embodiment of the present invention, as the organic solvent, a
water-miscible organic solvent, for example, selected from the
group consisting of alcohol (for example, ethanol), acetone,
tetrahydrofuran, acetic acid, acetonitrile and dioxane and
combinations thereof can be used, but it is not limited thereto. In
addition, as the aqueous solvent, one selected from the group
consisting of conventional water, distilled water, distilled water
for injection, physiological saline, 5% glucose, buffer and
combinations thereof can be used, but it is not limited
thereto.
[0060] The method for preparing a pharmaceutical composition of an
embodiment of the present invention may further comprise removing
an organic solvent after said step (a).
[0061] In an embodiment, the method may further comprise
lyophilizing the micelle composition with addition of a
lyophilization aid. The lyophilization aid may be added for the
lyophilized composition to maintain a cake form. In another
embodiment, the lyophilization aid may be one or more selected from
the group consisting of sugar and sugar alcohol. The sugar may be
one or more selected from lactose, maltose, sucrose or trehalose.
The sugar alcohol may be one or more selected from mannitol,
sorbitol, maltitol, xylitol and lactitol. The lyophilization aid
may also function to facilitate homogeneous dissolution of the
lyophilized polymeric micelle composition upon reconstitution. The
lyophilization aid may be contained at an amount of 1 to 90 weight
%, particularly, 1 to 60 weight %, more particularly 10 to 60
weight %, based in a total weight of the lyophilized
composition.
[0062] The present invention is explained in more detail by the
following examples. However, these examples seek to illustrate the
present invention only, and the scope of the present invention is
not limited by the examples in any manner.
EXAMPLES
Preparation Example 1
Synthesis of Diblock Copolymer Consisting of
Monomethoxypolyethylene Glycol and D,L-lactide (mPEG-PDLLA) and
Purification by Sublimation Method
[0063] 150 g of monomethoxypolyethylene glycol (mPEG, number
average molecular weight=2,000) was fed into a 500-ml round-bottom
flask equipped with an agitator, and agitated at 120.degree. C.
under vacuum condition for 2 hours to remove moisture. 0.15 g of
tin octoate (Sn(Oct).sub.2) dissolved in 200 .mu.l of toluene was
added in the reaction flask, and further agitated under vacuum
condition for 1 hour to distill and remove toluene. 150 g of
D,L-lactide was then added and agitated under nitrogen atmosphere
for dissolution. After D,L-lactide was dissolved completely, the
reactor was tightly sealed and the polymerization reaction was
conducted at 120.degree. C. for 10 hours. After the reaction was
terminated, under agitation with a magnetic bar, the reactor was
connected to a vacuum pump and the product was purified under a
pressure of 1 torr or less by a sublimation method for 7 hours to
obtain 262 g of mPEG-PDLLA in molten state. The molecular weight
(Mn: .about.3740) was calculated by analyzing with .sup.1H-NMR
obtaining relative intensities of appropriate peaks with reference
to --OCH.sub.3 which is the terminal group of
monomethoxypolyethylene glycol.
Preparation Example 2
Purification of Diblock Copolymer (mPEG-PDLLA) by Sublimation
Method
[0064] 30 g of mPEG-PDLLA, which was obtained in the polymerization
reaction process of Preparation Example 1 before conducting the
purification process, was fed into a one-necked flask and dissolved
at 80.degree. C. Under agitation with a magnetic bar, the reactor
was connected to a vacuum pump and the product was purified under a
pressure of 1 torr or less by a sublimation method for 24 hours and
48 hours.
Preparation Example 3
Purification of Diblock Copolymer (mPEG-PDLLA) by Sublimation
Method
[0065] Except that the purification temperature was 100.degree. C.,
the purification was conducted by the same method as in Preparation
Example 2.
Preparation Example 4
Purification of Diblock Copolymer (mPEG-PDLLA) by Sublimation
Method
[0066] Except that the purification temperature was 120.degree. C.,
the purification was conducted by the same method as in Preparation
Example 2.
Preparation Example 5
Purification of Diblock Copolymer (mPEG-PDLLA) by Adsorption Method
Using Aluminum Oxide (Al.sub.2O.sub.3)
[0067] 30 g of mPEG-PDLLA, which was obtained in the polymerization
reaction process of Preparation Example 1 before conducting the
purification process, was fed into a one-necked flask and dissolved
by adding acetone (60 ml). Aluminum oxide (15 g) was added thereto
and completely mixed. The one-necked flask was connected to a
rotary evaporator, and the contents were mixed at 50.degree. C. at
60 rpm for 2 hours. The solution was then filtered at room
temperature with PTFE filter paper (1 .mu.m) to remove aluminum
oxide. The filtered acetone solution was distilled using a rotary
evaporator at 60.degree. C. under vacuum to remove acetone, thereby
to obtain the purified mPEG-PDLLA. The molecular weight (Mn:
.about.3690) was calculated by analyzing with .sup.1H-NMR obtaining
relative intensities of appropriate peaks with reference to
--OCH.sub.3 which is the terminal group of monomethoxypolyethylene
glycol.
[0068] The molecular weight change of mPEG-PDLLA according to the
purification conditions in the above Preparation Examples 2 to 5 is
shown in the following Table 1.
TABLE-US-00001 TABLE 1 Purification Purification Molecular
Temperature (.degree. C.) Time (hr) weight (Mn) Preparation Example
2 80 24 3740 48 3740 Preparation Example 3 100 24 3720 48 3700
Preparation Example 4 120 24 3650 48 3550 Preparation Example 5
Al.sub.2O.sub.3 purification 3690
[0069] From the results of Table 1, it can be seen that the reduced
amount of the molecular weight of mPEG-PDLLA increases as the
purification temperature becomes higher. The purification condition
of 80 to 100.degree. C. and 24 to 48 hours, particularly
100.degree. C. and 24 hours, can be thought of as efficient.
Comparative Example 1
Preparation of Polymeric Micelle Composition Containing
Paclitaxel
[0070] 1 g of paclitaxel and 5 g of mPEG-PDLLA obtained in
Preparation Example 1 were weighed, and 4 ml of ethanol was added
thereto and agitated at 60.degree. C. until the mixture was
completely dissolved to form a clear solution. Ethanol was then
removed by distillation under reduced pressure using a rotary
evaporator equipped with a round-bottom flask at 60.degree. C. for
3 hours. The temperature was then lowered to 50.degree. C., and 140
ml of distilled water at room temperature was added and reacted
until the solution became clear in blue color to form polymeric
micelles. As a lyophilization aid, 2.5 g of anhydrous lactose was
added thereto and dissolved completely, filtered using a filter
with a pore size of 200 nm, and freeze-dried to obtain a polymeric
micelle composition containing paclitaxel in powder form.
Example 1
Preparation of Polymeric Micelle Composition Containing
Paclitaxel
[0071] Except that mPEG-PDLLA purified for 24 hours in Preparation
Example 3 was used, a polymeric micelle composition containing
paclitaxel was prepared by the same method as in Comparative
Example 1.
Example 2
Preparation of Polymeric Micelle Composition Containing
Paclitaxel
[0072] Except that mPEG-PDLLA purified in Preparation Example 5 was
used, a polymeric micelle composition containing paclitaxel was
prepared by the same method as in Comparative Example 1.
Experimental Example 1-1
Isolation of Related Compound by Liquid Chromatography
[0073] To a vial containing 100 mg of polymeric micelle composition
containing paclitaxel, which had been subjected to the six-month
acceleration test (temperature: 40.degree. C.), 16.7 ml of
deionized water (DW) was fed and the contents were completely
dissolved, and the total amount of the liquid was taken and
transferred to a 20-ml volumetric flask, and the marked line was
met to make the total volume 20 ml (5.0 mg/ml). 2 ml of this liquid
was taken and transferred to a 10-ml volumetric flask, and the
marked line was met with acetonitrile to make the total volume 10
ml (1 mg/ml). For the above composition, related compound was
isolated and fractionally collected using the following liquid
chromatography.
[0074] Conditions for liquid chromatography
[0075] 1) Column: Poroshell 120 PFP (4.6.times.150 mm, 2.7 .mu.m,
Agilent)
[0076] 2) Mobile phase: A: DW/B: Acetonitrile
TABLE-US-00002 Time (min) % A % B 0.00 65 35 25.00 45 55 28.00 45
55 30.00 65 35 35.00 65 35
[0077] 3) Flow rate: 0.6 ml/min
[0078] 4) Injection volume: 10 .mu.l
[0079] 5) Detector: UV absorption spectrophotometer (Measurement
wavelength: 227 nm)
[0080] The resulting chromatogram of HPLC analysis is shown in FIG.
1.
Experimental Example 1-2
Qualitative Analysis of Related Compounds Using LC/MS/MS
[0081] The related compounds isolated in Experimental Example 1-1
(RRT: 1.10.+-.0.02 (1.08.about.1.12) and 1.12.+-.0.02
(1.10.about.1.14)) were qualitatively analyzed by MS scan of liquid
chromatography-mass spectrometer (LC/MS/MS). As the LC/MS/MS,
liquid chromatography 1200 series and electrospray ionization mass
spectrometer 6400 series (Agilent, US) were used. The conditions
for analysis were as follows.
[0082] Conditions for Liquid Chromatography
[0083] 1) Column: Cadenza HS-C18 (3.0.times.150 mm, 3 .mu.m,
Imtakt)
[0084] 2) Mobile phase: A: 0.5 mM ammonium acetate with 0.03%
acetic acid/B: Acetonitrile
TABLE-US-00003 Time (min) % A % B 0.00 80 20 4.00 55 45 9.00 55 45
9.10 80 20 15.00 80 20
[0085] 3) Flow rate: 0.4 ml/min
[0086] 4) Injection volume: 2 .mu.l
[0087] 5) Detector: UV absorption spectrophotometer (Measurement
wavelength: 227 nm)
[0088] Conditions for Electrospray Ionization Mass Spectrometer
[0089] 1) Ionization: Electrospray Ionization, Positive (ESI+)
[0090] 2) MS Method: MS2 scan/Product ion scan
[0091] 3) Ion source: Agilent Jet Stream ESI
[0092] 4) Nebulizer gas (pressure): Nitrogen (35 psi)
[0093] 5) Ion spray voltage: 3500 V
[0094] 6) Drying gas temperature (flow rate): 350.degree. C. (7
L/min)
[0095] 7) Sheath gas temperature (flow rate): 400.degree. C. (10
L/min)
[0096] 8) Fragmentor: 135 V
[0097] 9) Nozzle voltage: 500 V
[0098] 10) Cell accelerator voltage: 7 V
[0099] 11) EMV: 0 V
[0100] 12) Collision energy: 22 V
[0101] 13) Precursor ion: m/z 1020.2
[0102] 14) Mass scan range: m/z 100.infin.1500
[0103] The substance for analysis, which was isolated and came out
of the detection stage, was set to flow in the mass spectrometer,
and at that time the detected ion of related compound was
qualitatively analyzed selecting the characteristic ion of mass
spectrum [M+Na].
[0104] The resulting spectrum of LC/MS/MS analysis is shown in FIG.
2.
Experimental Example 2
Induction of Reaction of Paclitaxel and Lactide and HPLC Analysis
of Reaction Product
[0105] In the related compounds which were fractionally collected
from the polymeric micelle composition containing paclitaxel in
Experimental Example 1-1, many polymers existed together and thus
direct experiment was very difficult. As a result of the
qualitative analysis through Experimental Example 1-2, the related
compounds at RRT 1.10 and 1.12 positions were presumed as compounds
produced by combination of paclitaxel and lactide. Accordingly, an
experiment of inducing a reaction by adding lactide to paclitaxel
directly and analyzing the reaction product was carried out to
confirm whether the presumed related compounds were produced.
[0106] First, each of 5 mg of paclitaxel and 3 mg of
L-lactide/D-lactide was dissolved in 1 ml of acetonitrile
(ACN):DW=70:30 (v/v) solution, and the solutions were then mixed.
This solution was transferred to an LC vial and analyzed by HPLC.
The resulting chromatogram of HPLC analysis is shown in FIG. 3.
[0107] As a result of analysis, the peaks which newly appeared from
the reaction product of L-lactide/D-lactide and paclitaxel were
exactly identical with the peaks of impurities shown at RRT 1.10
and 1.12 in HPLC analysis after the six-month acceleration test of
the polymeric micelle pharmaceutical composition. Together with
this, it could also be confirmed that the compound of the
combination of paclitaxel and L-lactide eluted first on HPLC and
the combination of paclitaxel and D-lactide eluted later.
Furthermore, in case of experiment using the same amount, it could
be confirmed that the combination of paclitaxel and D-lactide was
formed more than the combination of paclitaxel and L-lactide. It
could be known from this that the impurities appearing at RRT 1.10
and 1.12 positions from the polymeric micelle composition
containing paclitaxel, which had been subjected to the six-month
acceleration test, were the materials formed as a result of
reacting paclitaxel with L-lactide and D-lactide, respectively.
Experimental Example 3
Analysis of Reaction Product of Paclitaxel and Lactide Using
LC/MS/MS
[0108] Using LC/MS/MS, a sample containing paclitaxel only was MS
scanned first, and as a result thereof, m/z 854.2 amu which was
[M+H].sup.+ and m/z 876.2 amu which was [M+Na].sup.+ appeared.
After that, when L-lactide and D-lactide were added to paclitaxel,
m/z 1020.0 amu, which was not shown in the sample containing
paclitaxel only, appeared and it was confirmed that the intensity
thereof continuously increased with the lapse of time. The
resulting spectrum of LC/MS/MS analysis is shown in FIG. 4.
[0109] It could be confirmed again from this that the structure of
the related compound obtained through Experimental Example 1-1 was
the 1020.0 amu compound which was [M+N].sup.+ produced by the
combination of paclitaxel and lactide isomer.
[0110] From the results of Experimental Examples 2 and 3 and
conventional knowledge of reaction in organic chemistry, it could
be known that the related compound at RRT 1.10 and 1.12 of the
polymeric micelle composition was the following compound produced
by the combination of paclitaxel and (L/D-) lactide.
##STR00006##
[0111] Combined form of paclitaxel and lactide:
C.sub.53H.sub.59NO.sub.18 (998.03 g/mol)
Experimental Example 4
Comparative Test of Storage Stability of Polymeric Micelle
Containing Drug at Severe Condition (80.degree. C.)
[0112] The polymeric micelle compositions of paclitaxel prepared in
Comparative Example 1 and Examples 1 and 2 were kept in an oven at
80.degree. C. for 3 weeks, and the compositions were then analyzed
with HPLC to compare the amounts of related compound. The test
solution was prepared by dissolving the micelle composition in 80%
acetonitrile aqueous solution and diluting to 600 ppm concentration
of paclitaxel. The resulting chromatogram of HPLC analysis is shown
in FIG. 5 and the change in the amount of related compound (%)
according to the severe test time is shown in the following Table
2.
[0113] HPLC Conditions
[0114] Column: Diameter 2.7 .mu.m, poroshell 120PFP (4.6.times.150
mm, 2.7 .mu.m) (Agilent column)
[0115] Mobile Phase
TABLE-US-00004 Time (min) Water:Acetonitrile 0~25 65:35 .fwdarw.
45:55 25~28 45:55 28~30 45:55 .fwdarw. 65:35 30~35 65:35
[0116] Detector: UV absorption spectrophotometer (227 nm)
[0117] Flow rate: 0.6 ml/min
[0118] Amount of each related compound (%)=100(Ri/Ru)
[0119] Ri: Area of each related compound detected in test solution
analysis
[0120] Ru: Sum of all peak areas detected in test solution
analysis
TABLE-US-00005 TABLE 2 RRT* 0.87 .+-. 0.02 0.96 .+-. 0.02 1.10 .+-.
0.02 1.12 .+-. 0.02 1.44 .+-. 0.05 Sample-Storage time (0.85~0.89)
(0.94~0.98) 1.00 (1.08~1.12) (1.10~1.14) (1.39~1.49) Comparative
Example 1-0 day(d) 0.04% 0.03% 99.74% -- -- -- Comparative Example
1-3 weeks(w) 0.76% 1.12% 92.59% 0.18% 0.27% 0.93% Example 1-0
day(d) 0.03% 0.02% 99.64% 0.02% -- -- Example 1-3 weeks(w) 0.11%
0.15% 95.44% 0.06% 0.08% 0.22% Example 2-0 day(d) -- 0.02% 99.72%
-- -- -- Example 2-3 weeks(w) 0.40% 1.04% 93.94% 0.08% 0.05% 0.11%
*RRT 0.87 .+-. 0.02: Paclitaxel, oxetane ring opened compound RRT
0.96 .+-. 0.02: Paclitaxel, oxetane ring opened compound RRT 1.00:
Paclitaxel RRT 1.10 .+-. 0.02: Paclitaxel, L-lactide reaction
compound RRT 1.12 .+-. 0.02: Paclitaxel, D-lactide reaction
compound RRT 1.44 .+-. 0.05: Paclitaxel, water eliminated
compound
[0121] From Table 2 and FIG. 5, it can be known that the stability
of the polymeric micelle pharmaceutical composition of Example 1 or
2 was improved as compared with the composition of Comparative
Example 1 and the reduction of paclitaxel amount was relatively
smaller, whereby the effect of the drug contained in the
composition can be maintained more stably.
Experimental Example 5
Comparative Test of Storage Stability of Polymeric Micelle
Containing Drug at Accelerated Condition (40.degree. C.)
[0122] Except that the polymeric micelle composition of paclitaxel
prepared in Comparative Example 1 and Example 1 respectively was
kept in a stability tester at 40.degree. C. for 6 months, the test
was conducted by the same method as in Experimental Example 4. The
change in the amount of related compound (%) according to the
acceleration test time is shown in the following Table 3.
TABLE-US-00006 TABLE 3 RRT* Sample-Storage 0.87 .+-. 0.02 1.10 .+-.
0.02 1.12 .+-. 0.02 1.44 .+-. 0.05 time (0.85~0.89) (1.08~1.12)
(1.10~1.14) (1.39~1.49) Comparative 0.17% 0.22% 0.36% 0.12% Example
1-6 months Example 1-6 0.04% 0.05% 0.02% 0.04% months
[0123] The above test result shows an average value of the amounts
of each related compound and paclitaxel in the test conducted for 3
or more polymeric micelle compositions of different batches.
[0124] Through Experimental Example 5, it has been proven that the
composition of Example 1, if stored at the accelerated storage
temperature (40.degree. C.) for 6 months, has lower amount of
related compound than the composition of Comparative Example 1.
* * * * *