U.S. patent application number 14/284152 was filed with the patent office on 2014-11-13 for pharmaceutical composition containing docetaxel-cyclodextrin inclusion complex and its preparing process.
The applicant listed for this patent is Meridian Laboratories, Inc.. Invention is credited to Jianfeng Gao, Yong Ren, Ling Wu, Shuqin Yu.
Application Number | 20140336149 14/284152 |
Document ID | / |
Family ID | 37877491 |
Filed Date | 2014-11-13 |
United States Patent
Application |
20140336149 |
Kind Code |
A1 |
Ren; Yong ; et al. |
November 13, 2014 |
PHARMACEUTICAL COMPOSITION CONTAINING DOCETAXEL-CYCLODEXTRIN
INCLUSION COMPLEX AND ITS PREPARING PROCESS
Abstract
A docetaxel inclusion complex having improved water-solubility
(up to 5 mg/ml) and stability (stability constant
Ka=2056M.sup.-1-13051M.sup.-1), comprises docetaxel and
hydroxypropyl-beta-cyclodextrin and/or sulfobutyl-beta-cyclodextrin
in a ratio of 1:10-150. The method includes steps as follows:
docetaxel dissolved in ethanol is added into water solution of
cyclodextrin via stirring, until docetaxel is completely dissolved;
said solution is filtered in 0.2-04 .mu.m microporous membrane then
ethanol is removed through reduced pressure to obtain the inclusion
complex in a liquid form; or ethanol, followed by water is removed
through reduced pressure, then dried to obtain the inclusion
complex in a solid form.
Inventors: |
Ren; Yong; (Nanjing, CN)
; Gao; Jianfeng; (Nanjing, CN) ; Yu; Shuqin;
(Nanjing, CN) ; Wu; Ling; (Nanjing, CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Meridian Laboratories, Inc. |
Buffalo |
NY |
US |
|
|
Family ID: |
37877491 |
Appl. No.: |
14/284152 |
Filed: |
May 21, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13932434 |
Jul 1, 2013 |
8765716 |
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14284152 |
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12440942 |
Mar 12, 2009 |
8481511 |
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PCT/CN2006/002692 |
Oct 13, 2006 |
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13932434 |
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Current U.S.
Class: |
514/58 |
Current CPC
Class: |
A61K 31/337 20130101;
B82Y 5/00 20130101; A61K 47/6951 20170801; A61K 9/0019 20130101;
A61P 35/00 20180101 |
Class at
Publication: |
514/58 |
International
Class: |
A61K 47/48 20060101
A61K047/48; A61K 31/337 20060101 A61K031/337 |
Claims
1. A pharmaceutical composition comprising cyclodextrin/doectaxel
inclusion, which consists of docetaxel, cyclodextrin and a
pharmaceutically acceptable excipient, wherein the mass ratio of
the docetaxel to cyclodextrin is 1:10.about.150, the said
cyclodextrin is hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin or their mixture; the stability
constant of the cyclodextrin/docetaxel inclusion is
Ka=2056M.sup.-1-13051M.sup.-1.
2. The pharmaceutical composition according to claim 1, wherein,
the preparation method of said cyclodextrin/docetaxel inclusion
pharmaceutical composition is as follows: a solution of docetaxel
in ethanol is added dropwise to an aqueous solution of
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin or their mixture while stirring; the
resulting mixture is filtered through microporous membrane of
0.2.about.0.4 .mu.m after being dissolved; the ethanol is removed
from the filtrate under reduced pressure, then water is also
removed under reduced pressure, the resulting product is dried
giving a solid inclusion, or the ethanol is removed under reduced
pressure giving a liquid inclusion.
3. A preparation method of the pharmaceutical composition
comprising cyclodextrin/docetaxel inclusion according to in claim 1
is as follows: a solution of docetaxel in ethanol is added dropwise
to an aqueous solution of
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-3-cyclodextrin or their mixture while stirring; the
resulting mixture is filtered through microporous membrane of
0.2.about.0.4 .mu.m after being dissolved; the ethanol is removed
from the filtrate under reduced pressure giving a liquid inclusion,
or the ethanol is removed under reduced pressure, then the water is
removed under reduced pressure, the resulting product is dried
giving a solid inclusion.
4. The preparation method o according to claim 3, wherein, the
specific steps are as follows: the docetaxel and cyclodextrin in a
mass ratio of 1:17 are used to prepare an aqueous solution of
cyclodextrin; the solution prepared with
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin, or their mixture and 2.about.10
times of pure water (in weight) is added with a mixed solution
prepared by docetaxel and appropriate amount of ethanol while
heating and stirring at room temperature or at a temperature of
25.about.65.degree. C.; after the inclusion forms, filtered through
microporous membrane of 0.2.about.0.4 .mu.m; then the ethanol is
removed under reduced pressure, and then continues to reduce
pressure until water is removed. The resulting product is dried
under a vacuum condition to give a solid inclusion.
Description
[0001] The present invention relates to a pharmaceutical
composition containing hydroxypropyl-sulfobutyl-.beta.-cyclodextrin
or sulfobutyl-.beta.-cyclodextrin/docetaxel inclusion complex and
the preparation method thereof.
CROSS REFERENCE AND RELATED APPLICATIONS
[0002] This application is a continuation of U.S. patent
application Ser. No. 13/932,434, filed Jul. 1, 2013, which is a
continuation of U.S. patent application Ser. No. 12/440,942, filed
Mar. 12, 2009, now U.S. Pat. No. 8,481,511, which is a 371 national
phase filing of PCT/CN06/02692, filed Oct. 13, 2006. The entire
contents of each of the above-referenced applications are
incorporated herein by reference.
FIELD OF THE INVENTION
BACKGROUND OF THE INVENTION
[0003] Docetaxel (trade name: Taxotere) is a synthesized compound
after structural modification extracted from Taxus baccata leaves,
a white or almost white crystalline powder, with molecular formula
C.sub.43H.sub.53N.sub.14, chemical name:
[2aR-(2a.alpha.,4.beta.,4a.beta.,6.beta.,9.alpha.,(.alpha.R*,.beta.S*),11-
.alpha.,12.alpha.,12a.alpha.,12b.alpha.)]-.beta.-[[(1,1-dimethyl
ethoxy)carbonyl]amino]-.alpha.-hydroxyphenyl propionic acid
[12b-acetoxy-12-benzoyloxy-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecahydro-4-
,6,11,trihydroxyl-4a,8,13,13-tetramethyl-5-oxo-7,11-methylene-1H-pentacycl-
o-ene[3,4]benzo[1,2-b]tetraoxacyclo-9-base]acetate trihydrate. Its
molecular weight is 807.19. This product is a new taxus antitumor
drug, belongs to a fat-soluble compound, difficult to dissolve in
water (2.903 .mu.g/ml). Because the benzoyl group of the paclitaxel
is substituted by t-butoxycarbonyl, the water-solubility of
Docetaxel is slightly larger than that of Paclitaxel (0.25
.mu.g/ml.about.0.60 .mu.g/ml).
[0004] The anti-tumor mechanism of this product is the same as that
of paclitaxel, which exerts its anti-tumor activity by inhibiting
tubulin depolymerization. However, its inhibitory activity of
tubulin depolymerization is about 2 times of that of paclitaxel, so
it has a better curative effect on advanced breast carcinoma,
superior to other single drug therapy (25%-53%) and stronger than
anthracyclines, with an effective ratio of 49% for treatment of
advanced breast carcinoma and better curative effects on advanced
non-a-cell lung cancer, advanced ovarian cancer, pancreatic cancer,
head and neck cancer and gastric cancer.
[0005] Docetaxel is insoluble in water and other commonly used
medicinal organic solvent. It is used as injection in clinical
application, and its injection solution is prepared with docetaxel
Tween solution (injection specification: 1 ml, 20 mg/ml) and 13%
ethanol and normal saline or glucose. The tween solution has a
lower stability, stored at a low temperature of 2.about.8.degree.
C. in a dark place. The injection solution for clinical use is
easily to precipitate. This product has similar side effects as
paclitaxel, which may cause serious allergic reactions after
injection, and the incidence rate of moderate and severe
hypersusceptibility is as high as 25.9% (Onconlogy, 1997,11 (7):
11); If the dosage is 75 mg/m.sup.2 or 100 mg/m.sup.2, the
incidence rate of moderate hypersusceptibility or more is
respectively 31% and 41.3% and the injection-part reaction rate can
also be up to 13.3% (Cancer, 1995,754:968). Therefore, when
docetaxel is used, allergic reaction prevention treatment should be
conducted by taking large doses of hormones from PM8:00 on the
first day before the docetaxel treatment, generally taking
dexamethasone 4.5 mg orally, once in the morning and once at night
every day, for 3.about.5 days in succession; 30 min-60 min before
injection of docetaxel, intramuscular injection of 40 mg
diphenhydramine and 300 mg cimetidine were conducted to prevent
from hypersensitivity. To prevent gastrointestinal reaction,
intravenous injection of ondansetron or Nasea shall be
conventionally conducted 30 min prior to chemotherapy In addition,
it is quite troublesome for clinical use of docetaxel, closely
observing whether there are teaks or not in the process of fluid
infusion; if any, the injection part must be immediately replaced
and local blocking should be conducted with Hirudoid ointment for
external use (3 times per day for a few days). In addition, the
conventional ECG monitoring should be carried out to closely
observe the changes of breathing, heart rate and blood pressure,
pay close attention to the occurrence of allergic reactions and
make sure a good preparation of care rescue for severe allergic
reactions. At present, improving the stability of docetaxel
formulations and reducing the side effects become the focus of
technical difficulties.
[0006] In recent years, researches on paclitaxel formulations
mainly focus on liposomes, nano-granule, albumin cross-linked
precursor, cyclodextrin inclusion and so on. The key technology,
focuses on biocompatibility, in vivo tolerance of the selected
materials, paclitaxel solubilization and formulation stability.
Similar to paclitaxel, for the docetaxel formulation improvement,
the cyclodextrin inclusion technology is increasing widely used,
and acet-.gamma.-cyclodextrin, Hydroxypropyl-.gamma.-cyclodextrin
and 2,6-dimethyl-.beta.-cyclodextrin have been reported to use for
docetaxel modification. Current study showed that application of
cyclodextrin inclusion technology can not only enhance docetaxel
stability, enhance its solubility, but also can significantly
enhance the drug activity and reduce toxic and side effects. By
using methyl-.beta.-cyclodextrin/docetaxel inclusion and other
drugs to determine their effects on the activity of a wide range of
tumor cells IC.sub.50 (European Journal of Cancer, 1998, 34:
168-174), it was discovered that, after adding
methyl-.beta.-cyclodextrin, docetaxel IC.sub.50 was significantly
reduced and IC.sub.50/IC.sub.50CD values f different cells were
within the scope ranging from 4.7-14.3, which proved that,
cyclodextrin inclusion can significantly enhance the activity of
docetaxel. The inclusion formulations made of
acet-.beta.-cyclodextrin or hydroxypropyl-.beta.-cyclodextrin
(WO9924073; CN1222321/ZL 98811010.5) can significantly reduce toxic
and side effects of cardiovascular and respiratory system, but
preferable technical solution in the invention is to prepare the
composition with docetaxel/hydroxypropyl-.beta.-cyclodextrin in a
mass ratio of 1:133, the solubility of docetaxel is 0.75 mg/ml; if
with the docetaxel/acetyl-.gamma.-cyclodextrin in a mass ratio
1:50; the solubility of docetaxel can be up to 1.00 mg/ml, despite
of increased docetaxel solubility, but each dose specification (20
mg) of docetaxel injection solution prepared by such technology
still require not less than 1 ml ethanol as a cosolvent. If the
existing technology is used to conduct cyclodextrin docetaxel
inclusion, which would be unable to meet the drug requirements for
clinical use, the main problem is: The solubility of docetaxel
after cyclodextrin inclusion is still relatively low inconvenient
for clinical use (need substantial solvent) and difficult to meet
the suitable dosage requirements; application of substantial
cyclodextrin will seriously affect practical applications;
solubilization of inclusion with high doses of organic solvent can
not only be conducive to cyclodextrin solubilization, but also
increase the formulation irritation and reduce the patient's
compliance. In aspect of technical methods, current technology only
focuses on pharmaceutical stability of formulations
(clarification/precipitation of the solution), while ignoring the
drug chemical stability of especially for in the solution. Because
cyclodextrin has the features of catalyzing ester compounds
(Organic Chemistry, 2002; 22 (11):827-834), docetaxel containing
ester-based side chains in solution could tend to stabilize due to
cyclodextrin inclusion, or accelerate decomposition by cyclodextrin
catalysis (cause decrease of stability). The inclusion is directly
related to the stability and cyclodextrin structural property (type
of cyclodextrin). These factors caused difficult extended
application of current technology. Therefore, less use of
cyclodextrin and no use or less use of organic solvents for
improving the docetaxel solubility to meet the dose requirements of
clinical application and maintaining the drug pharmaceutical nature
and chemical stability are the technical problems for use of
cyclodextrin inclusion technology in docetaxel.
[0007] At present, the cyclodextrin and cyclodextrin derivatives
used to injection only include .alpha.-cyclodextrin,
hydroxypropyl-.beta.-cyclodextrin and
sulfobutyl-.beta.-cyclodextrin only (Expert Opin Drug Deliv, 2005
March; 2 (2): 335-51), among of which,
hydroxypropyl-.beta.-cyclodextrin is a neutral non-ionized
derivative and sulfobutyl-.beta.-cyclodextrin is an ionized
derivative. Studies shows that (Medicine Forefront, Chinese
Medicine Science and Technology Press, 2001: 46-59),
sulfobutyl-.beta.-cyclodextrin was significantly improved in
safety, stability, drug solubilization and production preparation
technology, etc. Recently, we researched and developed a new type
of cyclodextrin
derivative-hydroxypropyl-sulfobutyl-.beta.-cyclodextrin (CN
1800221A), which has excellent performance and good safety. In the
present invention, hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin was used to improve docetaxel. The
low proportion (mass ratio 1:17) of inclusion can promote the
docetaxel solubility up to 15 mg/ml or higher. The prepared
injection has a good pharmaceutical property and chemical
stability, strong activity and low irritation. If the solid
inclusion was diluted for 500 times, it can maintain stable for
several days, with an important clinical application value.
SUMMARY OF THE INVENTION
[0008] The present invention is to overcome the drawbacks of low
docetaxel solubility, poor pharmaceutical chemical stability; too
high cyclodextrin ratio and much residual organic solvents, realize
technological breakthroughs and provide a cyclodextrin docetaxel
inclusion featuring in high docetaxel solubility and pharmaceutical
stability, relatively low cyclodextrin content and small residual
organic solvents so as to realize clinical application. The present
invention also provides the preparation method of such inclusion
composition.
[0009] For the present invention, the pharmaceutical composition
containing cyclodextrin/docetaxel inclusion composes docetaxel,
cyclodextrin and pharmaceutical acceptable excipient. Of which, the
mass ratio of docetaxel and Cyclodextrin is 1:10.about.150; said
cyclodextrin is hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin or their mixture; the stability
constant of the cyclodextrin/docetaxel inclusion is
Ka=2056M.sup.-1.about.13051M.sup.-1.
[0010] Specifically, the said pharmaceutical composition comprising
cyclodextrin/docetaxel inclusion is prepared as follow:
[0011] Docetaxel dissolved in ethanol is added into a water
solution of hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin or their mixture while stirring;
[0012] Until docetaxel is completely dissolved, the resulting
solution is filtered in 0.2.about.0.4 .mu.m microporous
membrane;
[0013] Then ethanol is removed through reduced pressure to obtain
the inclusion complex in a liquid form;
[0014] Or ethanol after water is removed through reduced pressure,
then dried to obtain the inclusion complex in a solid form. The
resulting inclusion contains small or trace amount of ethanol. Said
"small or trace amount of ethanol" means that the content of
ethanol in the solid inclusion is less than 2%.
[0015] In the present invention,
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin aqueous solution was mixed with
proper amount of docetaxel in ethanol. The ethanol can promote the
inclusion. After inclusion is formed, the ethanol is removed to
prepare a stable docetaxel/cyclodextrin inclusion. The composition
of the inclusion and common pharmaceutical excipients was prepared
into a pharmaceutical inclusion composition for clinical use, thus
improving the docetaxel solubility, enhancing stability and
reducing the side effects, to obtain docetaxel products for
clinical applications. Preparation of inclusion is the key
technology of the present invention.
[0016] The sulfobutyl-.beta.-cyclodextrin used in the present
invention is ionized cyclodextrin derivative and the pharmaceutical
sulfobutyl-.beta.-cyclodextrin is a derivative(product) of
6.about.7 substitutions (SBE.sub.7-.beta.-CD trade name: Captisol);
and our developed hydroxypropyl-sulfobutyl-.beta.-cyclodextrin
(HP-SBE-.beta.-CD) is a new type of cyclodextrin derivative (CN
1800221A) after substituted by hydroxypropyl and sulfobutyl
combination, and this product has low toxicity, excellent
performance inclusion superior to sulfobutyl-.beta.-cyclodextrin,
more suitable for non-oral preparations.
[0017] Under the present invention, the preparation method of
pharmaceutical composition containing cyclodextrin/docetaxel
inclusion is as follows:
[0018] Docetaxel is prepared into a solution with less ethanol as
possible. Docetaxel dissolved in ethanol is added into a pure water
solution of hydroxypropyl-sulfobutyl-.beta.-cyclodextrin or
sulfobutyl-.beta.-cyclodextrin or their mixture while stirring;
[0019] Until docetaxel is completely dissolved, the resulting
solution is filtered in 0.2.about.0.4 .mu.m microporous
membrane;
[0020] The ethanol is removed through reduced pressure giving a
liquid inclusion, or ethanol after water is removed through reduced
pressure, then dried to obtain the docetaxel inclusion complex in a
solid form with the ethanol content of less than 2%.
[0021] More preferably and specifically, the detailed steps for
preparation of pharmaceutical composition containing
cyclodextrin/docetaxel inclusion are as follows:
[0022] The docetaxel and cyclodextrin in a weight ratio of 1:17
were used to prepare pure aqueous solution of cyclodextrin. The
solution prepared with
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin, or
sulfobutyl-.beta.-cyclodextrin, or the mixture and 2.about.10 times
of pure water (in weight) was added with the mixed solution
prepared by appropriate amount of docetaxel and ethanol (the
ethanol amount depending on dissolution of docetaxel), while
heating and stirring at room temperature or at a temperature of
25.about.65.degree. C. After the inclusion formed, filtered through
microporous membrane of 0.2.about.0.4 um; then the ethanol was
removed under reduced pressure to give a docetaxel solution
containing small amount or trace amount of ethanol.
[0023] The water is removed from the aforesaid docetaxel solution
through reduced pressure, and then dried under a vacuum condition
through reduced pressure to obtain the solid docetaxel inclusion
complex.
[0024] The resulting solid inclusion has a high water-solubility,
easily dissolved without adding other cosolvents. The aqueous
solution prepared has less side effect of hemolysis, suitable for
clinical use. The solid inclusion containing 20 mg of docetaxel
clinical dosage was added with 10-1000 times of injection saline
solution to prepare a solution, which could remain stable after a
few days. And the other isotonic dose of pharmaceutical excipient
added will basically not affect the stability of the solution.
[0025] The said solid inclusion and commonly-used injectable
pharmaceutical excipient solution were diluted to appropriate
concentration before subjecting to sterilization treatment. The
resulting composition solution can be used for injection.
Validation of Inclusion Reaction:
[0026] Determination of system UV absorption is an effective way to
identify inclusion (Cyclodextrin Chemistry, Science Press 2001,
P135). The UV Spectroscopy test showed that, in aqueous solution,
the UV absorption of docetaxel would vary with cyclodextrin
concentration (no Cyclodextrin absorption itself) (FIG. 1).
Docetaxel UV absorption would significantly enhance with the
increase of cyclodextrin concentration, suggesting that docetaxel
has a significantly strong inclusion with the cyclodextrin under
the present invention.
Determination of Inclusion Stability Constant:
[0027] The inclusion stability constant Ka is used to determine the
degree of inclusion and UV spectrophotometry is one of common
methods to determine Ka. With the change of cyclodextrin
concentrations, the UV absorption of docetaxel solution of constant
concentration shows a regular change, and the relationship between
cyclodextrin concentration (C) and UV absorbance (A) is obtained.
From the relation curve of 1/C and 1/A, the Level-1 apparent
stability constant Ka of cyclodextrin/docetaxel inclusion can be
quantitatively calculated. Meanwhile, the changes of Ka after
adding ethanol are determined in the test, and the Ka values of
different cyclodextrins are shown in Table 1.
TABLE-US-00001 TABLE 1 The Level-1 apparent stability constant Ka
of cyclodextrin/docetaxel inclusion(M.sup.-1; 231 nm) Ka
Cyclodextrin H.sub.2O 60% EtOH Sulfobutyl-.beta.-cyclodextrin 2056
6968 Hydroxypropyl-sulfobutyl-.beta.-cyclodextrin 13051 30117
Hydroxypropyl-.beta.-cyclodextrin 582 2694 .beta.-cyclodextrin
379
[0028] The results showed that in docetaxel inclusion, the
inclusion constant of cyclodextrin and
hydroxypropyl-.beta.-cyclodextrin is less than that of cyclodextrin
used in the present invention, this may explain why the effects of
prior arts are relatively poorer for the two cyclodextrins.
Cyclodextrin used in the present invention has a larger inclusion
constant Ka, and relevant test also showed that the presence of
ethanol can significantly enhance the value of Ka and enhance the
cyclodextrin inclusion ability, thus promoting the inclusion. The
ethanol is not just used as a cosolvent in the inclusion process.
Generally, when organic solvents compete with the drugs for
inclusion, ethanol would reduce the Ka value of drugs (Journal of
China Pharmaceutical University, 2005, 36 (1): 13-17). Unlike
Hydroxypropyl-.beta.-cyclodextrin, Cyclodextrin used in the present
invention still had a higher Ka value after ethanol was removed in
the pure water condition, enough to form a stable inclusion
complex. Therefore, the ethanol was removed as far as possible to
obtain a more pure drug inclusion and reduce the impact of ethanol
on the drugs as far as possible. For the present invention, the
impact of ethanol on the docetaxel inclusion is the technological
basis for preparation of inclusion.
Preparation and Validation of Inclusion:
[0029] After adding appropriate amount of ethanol to make full
inclusion, the ethanol was removed through reduced pressure, dried
to prepare different proportions of docetaxel/cyclodextrin
inclusion. DTA test confirmed that the solid substance is inclusion
rater than simple physical mixture. Taking
sulfobutyl-.beta.-cyclodextrin/docetaxel (mass ratio of 1:17) as an
example, it is analyzed and described as follows:
[0030] Four samples of docetaxel, sulfobutyl-.beta.-cyclodextrin,
physical mixture of docetaxel and sulfobutyl-.beta.-cyclodextrin
and inclusion were weighed, each for about 5.0 mg, to conduct
differential scanning thermal analysis: reference:
Al.sub.2O.sub.3range: .+-.50 .mu.V, temperature rise range:
30.degree. C..about.400.degree. C., heating rate 10.degree. C./min,
and the DTA pattern was obtained. The results showed that: for
docetaxel, a melting peak at 230.degree. C. and endothermic
decomposition at about 350.degree. C.; for sulfobutyl-cyclodextrin,
there is one dehydration endothermic peak and one phase transition
peak at 70-90.degree. C. and 250-270.degree. C. respectively and
melting endothermic decomposition at about 360.degree. C. For the
physical mixture, the endothermic peak characteristics of
cyclodextrin and docetaxel maintained, while for the inclusion, the
dehydration endothermic peak and phase transition peak basically
disappeared and a new phase transition peak appeared at about
180.degree. C. and exothermic decomposition at 360.degree. C., and
the position (temperature) and shape (thermal effect) of all peaks
were significantly changed, therefore, it proved that the inclusion
formed.
Residual Ethanol in Inclusion:
[0031] Inclusion.sup.1HNMR showed a weak ethanol methyl peak.
According to the integral area ratio of ethanol methyl peak (t,
.delta.=1.10912) and cyclodextrin characteristic peak (H1 peak, d,
.delta.=5.18257.about.5.05401), the residual ethanol amount (%) in
inclusions prepared was calculated, the results were shown in Table
2,
TABLE-US-00002 TABLE 2 Residual ethanol level of
docetaxel/cyclodextrin solid inclusion Residual Inclusion
Drug/Cyclodextrin volume batch No. Cyclodextrin * Ethanol (%)
20060111 a 1:17 0.17 20060121 b 1:17 0.13 20060221 a 1:30 0.32
20060302 b 1:30 0.27 20060513 b 1:50 0.30 20060511 a 1:50 0.44
20060408 a + b 1:25:25 0.36 20060623 b 1:100 1.15 * a =
Sulfobutyl-.beta.-cyclodextrin; b =
Hydroxypropyl-sulfobutyl-.beta.-cyclodextrin
[0032] The test showed that, although more ethanol was used in the
preparation process, there was less residual ethanol after
purification. For the inclusion prepared with a low proportion of
cyclodextrin (less than 1:50), generally the residual ethanol level
was less than 1.0%; even for the inclusion prepared with a higher
proportion of cyclodextrin, the residual ethanol level was less
than 2.0%. Low residual ethanol level provided a favorable
guarantee for improving the docetaxel stability and reducing
irritation and other side effects.
[0033] Inclusion Solubilization Test:
[0034] Standard curve: 0.67 mg/ml docetaxel solution as prepared by
dissolving docetaxel with ethanol, and diluted with pure water into
3.35 .mu.g/ml.about.20.1 .mu.g/ml series solutions, then UV
absorption value A was measured under 228 nm. A standard curve was
plotted by A against concentration C (mg/ml) (A=13.881C+0.3577
(r=0.9998)).
[0035] The excessive docetaxel or inclusion in the pure water was
oscillated for 72 h at 25.degree. C..+-.1.degree. C., filtered and
placed still. Appropriate amount of filtrate was fetched and
diluted with pure water, then absorbance intensity values were
measured at 228 nm to calculate the docetaxel solubility at
25.degree. C. from the standard curve, the results were shown in
Table 3.
TABLE-US-00003 TABLE 3 Docetaxel solubility in cyclodextrin
solution Cyclodextrin * Solubility Solubilization (drug proportion)
(mg/ml) Multiples 0 0.002903 1 a (1:17) 15.007 5169 b (1:17) 15.547
5355 * a = Sulfobutyl-.beta.-cyclodextrin; b =
Hydroxypropyl-sulfobutyl-.beta.-cyclodextrin
[0036] The results showed that cyclodextrin used in the present
invention had a strong impact on docetaxel solubilization and the
two cyclodextrins show insignificant difference on the docetaxel
solubilization. Significant enhancement of docetaxel solubility was
conducive to the preparation of stable non-oral preparations.
Sample Stability Test:
[0037] The pharmaceutical composition stability includes chemical
stability of drug and pharmaceutical stability of composition.
Solid Inclusion Stability
[0038] HPLC chromatographic conditions: chromatographic column ODS
C.sub.18(250 mm.times.4.6 mm); mobile phase:water:acetonitrile
(53:47); flow rate: 1.0 ml/min; detection wavelength: 230 nm;
detection time: 30.00 mm; detection sensitivity: 1.0000 AUFS.
[0039] Samples: docetaxel material;
docetaxel/sulfobutyl-.beta.-cyclodextrin inclusion (a, mass ratio
of 1:17); docetaxel/hydroxypropyl sulfobutyl-.beta.-cyclodextrin
inclusion (b, mass ratio of 1:17). Docetaxel materials and
inclusions were divided into three portions of test samples equally
and were subject to light, high temperature and high humidity
acceleration tests respectively:
(1) Light Test
[0040] Samples were put in a sealed transparent container and then
placed in an illumination box equipped with fluorescent light of
4500.+-.500LX illumination intensity for 5 days. Sampling
inspection analysis was conducted, and the test results were
compared with the samples of 0 day.
(2) High Temperature Test
[0041] Samples were placed in a clean sealed container at a
temperature of 60.degree. C. for 5 days. Sampling inspection
analysis was conducted, and the test results were compared with the
samples of 0 day.
(3) High Humidity Test
[0042] Samples were placed in a sealed container of constant
humidity at temperature of 25.degree. C. and relative humidity of
90.+-.5% for 5 days. Sampling inspection analysis was conducted,
and the test results were compared with the samples of 0 day.
[0043] HPLC determination charts see FIG. 2 and FIG. 3. All test
results see Table 4,
TABLE-US-00004 TABLE 4 Sample Content Determination of 5-day
Acceleration Test Content (%) High Hight Sample 0 day Light
temperature humitdity Docetaxel raw 99.675 90.176 87.859 83.877
material Inclusion a 99.684 96.478 94.235 95.331 Inclusion b 99.645
97.985 95.407 95.310
[0044] Under the acceleration test conditions, docetaxel raw
materials became slightly darker in color and the level decreased
significantly; the inclusion appearance presented no color change,
and the level decreased slightly and the impurities were basically
unchanged. The results showed that, for the solid inclusion, the
chemical property of docetaxel is stable and the inclusion
technology had a significant effect on improving the stability of
docetaxel.
Inclusion Solution and Pharmaceutical Stability
(1) Solution Stability
[0045] A solid inclusion with mass ratio of 1:17 was dissolved into
a liquid containing docetaxel of 15 mg/ml solution with normal
saline or isotonic glucose solution, then diluted 1.about.1000-fold
after sterilization treatment to prepare into different
concentrations of injections observed for 5 h-10 d in succession.
The observation results of
docetaxel/hydroxypropyl-sulfobuty-.beta.-cyclodextrin inclusion
saline dilution system stability were shown in Table 5.
TABLE-US-00005 TABLE 5 State of cyclodextrin/docetaxel inclusion
injection solution Drug Dilution concnetration Solution: State*
multiple mg/ml 0 h 5 h 10 h 15 h 20 h 2 d 4 d 6 d 10 d 1 7.50 + + +
+ + - - - - 10 1.50 + + + + + + + + + 20 0.75 + + + + + + + + + 50
0.30 + + + + + + + + + 100 0.15 + + + + + + + + + 500 0.03 + + + +
+ + + + + 1000 0.015 + + + + + + + + + *+: Clarification without
precipitation; -: precipitation or turbidity
(2) Docetaxel Stability Test in Solution (Assay)
[0046] Under the aforesaid HPLC chromatographic conditions, some
docetaxel raw materials; docetaxel/sulfobuty-.beta.-cyclodextrin
inclusion (a) with a mass ratio of 1:17 and
docetaxel/hydroxypropy-sulfobuty-.beta.-cyclodextrin inclusion (b)
with a mass ratio of 1:17 were taken and dissolved with normal
saline solution, diluted for 100 times after 30 min ultrasonic wave
treatment, placed still, and then the samples were taken to measure
HPLC chromatogram at 0 h,2 h,4 h,6 h,8 h. Thus the docetaxel
content in solution was obtained and the results were shown in
Table 6.
TABLE-US-00006 TABLE 6 Changes of sample content in aqueous
solution with time Content (%) Sample 0 h 5 h 10 h 24 h 96 h
Docetaxel 99.675 99.025 97.381 93.433 86.384 material Inclusion a
99.684 99.239 98.359 98.035 95.812 Inclusion b 99.645 99.554 99.002
97.459 96.386
[0047] Docetaxel was obviously decomposed, while the inclusion
samples had stable chemical properties in aqueous solution and its
content will remain basically unchanged within 10 hours. The
half-life of sample decomposition in solution was as follows:
362.413 hours for docetaxel, 1341.035 hours for inclusion a and
1502.927 hours for inclusion b. The inclusion technology enhanced
the docetaxel stability by 3.7 times and 4.15 times
respectively.
Hemolysis Test:
[0048] (References: Guidelines for Research Techniques of Chemicals
Stimulation, Hypersensitivity and Hemolysis Tests, State Food and
Drug Administration, Mar. 18, 2005; Technical requirements of
traditional Chinese medicine injection study", State Food and Drug
Administration, 1999, 11, 12)
[0049] There is significant difference of the hemolysis between the
inclusion saline solution and existing docetaxel formulations. The
diluted solution of sulfobutyl-.beta.-cyclodextrin inclusion (a)
has a slight hemolysis, while the
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin inclusion (b) has
lower hemolysis, within the range of clinical administration
concentration (about 0.08 mg/ml docetaxel content), there is
basically no hemolysis, while the hemolysis of the existing
docetaxel preparations is more than 20%. The results see FIG.
4.
ADVANTAGES OF THE INVENTION
[0050] The inclusion significantly improves the solubility of
docetaxel (up to 15 mg/ml and above). After dilution, the solution
can maintain clear and stable for a long time, with low hemolysis,
less side effect and good pharmaceutical activity.
[0051] After inclusion of docetaxel and cyclodextrin, the solid and
liquid samples have stable content, small proportion of
drug/cyclodextrin, low dosage of cyclodextrin in the formulation,
suitable for clinical use.
[0052] The inclusion in the present invention has less residual
organic solvents, conducive to improve the medication safety.
[0053] The preparation method is simple, easy to operate, low cost
without environmental pollution. The inclusion has stable property,
good compatibility with other pharmaceutical excipients, suitable
for preparation of formulations.
[0054] The injections prepared with inclusions contain no corrosive
ingredients, no poisons, convenient for clinical use and high
practicability.
DESCRIPTION OF DRAWINGS
[0055] FIG. 1: Docetaxel UV absorption scanning (220 nm.about.245
nm in aqueous solutions at different
hydroxypropyl-.beta.-cyclodextrin concentrations;
[0056] FIG. 2: HPLC chromatogram of
docetaxel/hydroxypropyl-sulfobutyl-.beta.-cyclodextrin inclusion
high temperature test (5 d);
[0057] FIG. 3: HPLC chromatogram of docetaxel raw material high
temperature test (5 d);
[0058] FIG. 4: Commercial docetaxel formulations,
docetaxel/hydroxypropyl-sulfobutyl-.beta.-cyclodextrin inclusion
and docetaxel/sulfobutyl-.beta.-cyclodextrin inclusion solutions
diluted with normal saline solution and docetaxel
concentration--hemolysis curve.
DETAILED DESCRIPTION OF THE INVENTION
EXAMPLE 1
[0059] 1.70 g hydroxypropyl-sulfobutyl-.beta.-cyclodextrin was
mixed with 5.0 ml pure water, then added dropwise slowly with a
solution prepared by 100.0 mg docetaxel and 1.0 ml ethanol while
stirring. After fully mixed until complete dissolution, the
resulting mixture was filtered through microporous membrane of
0.2.about.0.4 .mu.m. The ethanol was removed from the filtrate at
55.degree. C., under reduced pressure. After sterilization
treatment, the water was also removed under reduced pressure until
dried, and then the resulting solid product was dried for 48 h
under reduced pressure giving a white solid inclusion.
[0060] The above powdered inclusion 360 mg (containing 20 mg
docetaxel) was mixed and dissolved with 250 ml of normal saline
injection to a liquid inclusion composition before used for
injection.
EXAMPLE 2
[0061] The procedure was carried out basically the same as example
1, but wherein 3.0 g hydroxypropyl-sulfobutyl-.beta.-cyclodextrin
was mixed with 15 ml pure water. The weight of docetaxel was 20
mg.
EXAMPLE 3
The procedure was carried out basically the same as example 1, but
wherein, the weight of docetaxel was 170 mg.
EXAMPLE 4
[0062] The procedure was carried out basically the same as example
1, but wherein, the cyclodextrin was
sulfobutyl-.beta.-cyclodextrin.
EXAMPLE 5
[0063] The procedure was carried out basically the same as example
2, but wherein, the cyclodextrin was
sulfobutyl-.beta.-cyclodextrin.
EXAMPLE 6
[0064] The procedure was carried out basically the same as example
3, but wherein, the cyclodextrin was
sulfobutyl-.beta.-cyclodextrin.
EXAMPLE 7
[0065] The procedure was carried out basically the same as example
1, but wherein, the cyclodextrin was the mixture of
sulfobutyl-.beta.-cyclodextrin and
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin in a mass ratio of
1:1.
EXAMPLE 8
[0066] The procedure was carried out basically the same as example
2, but wherein, the cyclodextrin was the mixture of
sulfobutyl-.beta.-cyclodextrin and
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin in a mass ratio of
1:50.
EXAMPLE 9
[0067] The procedure was carried out basically the same as example
3, but wherein, the cyclodextrin was the mixture of
sulfobutyl-.beta.-cyclodextrin and
hydroxypropyl-sulfobutyl-.beta.-cyclodextrin in a mass ratio of
50:1.
EXAMPLE 10
[0068] The procedure was carried out basically the same as example
1, but, cyclodextrin and docetaxel were mixed with pure water
firstly, added dropwise with ethanol slowly until the system was
completely dissolved.
EXAMPLE 11
[0069] The procedure was carried out basically the same as example
1, but, the resulting powdered inclusion was diluted with isotonic
concentrations of glucose injection.
EXAMPLE 12
[0070] The procedure was carried out basically the same as example
1, but, the resulting powdered inclusion was diluted with isotonic
concentrations of fructose injection.
* * * * *