U.S. patent application number 13/825875 was filed with the patent office on 2014-03-13 for crystals of substituted cycloalkene derivatives.
This patent application is currently assigned to DAIICHI SANKYO COMPANY, LIMITED. The applicant listed for this patent is Kouki Iida, Takayoshi Nagasaki, Nobuyuki Ohkawa. Invention is credited to Kouki Iida, Takayoshi Nagasaki, Nobuyuki Ohkawa.
Application Number | 20140073689 13/825875 |
Document ID | / |
Family ID | 45873931 |
Filed Date | 2014-03-13 |
United States Patent
Application |
20140073689 |
Kind Code |
A9 |
Ohkawa; Nobuyuki ; et
al. |
March 13, 2014 |
Crystals of Substituted Cycloalkene Derivatives
Abstract
To provide potassium salts of substituted pyrrolidine
derivatives, which suppress the production of an inflammatory
mediator caused by intracellular signal transduction or cell
activation induced by endotoxin, and crystals thereof. The present
invention provides
potassium(2-chloro-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis-
(hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
and
potassium(2-bromo-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis(-
hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide,
each of which suppresses the production of an inflammatory mediator
caused by intracellular signal transduction or cell activation
induced by endotoxin, crystals thereof, and a pharmaceutical
containing any of the same, and a prophylactic and/or therapeutic
agent for sepsis containing any of the same.
Inventors: |
Ohkawa; Nobuyuki; (Tokyo,
JP) ; Iida; Kouki; (Tokyo, JP) ; Nagasaki;
Takayoshi; (Tokyo, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Ohkawa; Nobuyuki
Iida; Kouki
Nagasaki; Takayoshi |
Tokyo
Tokyo
Tokyo |
|
JP
JP
JP |
|
|
Assignee: |
DAIICHI SANKYO COMPANY,
LIMITED
Tokyo
JP
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20130178521 A1 |
July 11, 2013 |
|
|
Family ID: |
45873931 |
Appl. No.: |
13/825875 |
Filed: |
September 22, 2011 |
PCT Filed: |
September 22, 2011 |
PCT NO: |
PCT/JP2011/071557 PCKC 00 |
371 Date: |
March 25, 2013 |
Current U.S.
Class: |
514/467;
549/342 |
Current CPC
Class: |
A61P 43/00 20180101;
A61K 31/357 20130101; A61P 31/12 20180101; C07D 317/72 20130101;
A61P 31/04 20180101; A61P 29/00 20180101; A61P 9/00 20180101 |
Class at
Publication: |
514/467;
549/342 |
International
Class: |
C07D 317/72 20060101
C07D317/72 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 24, 2010 |
JP |
2010-213236 |
Claims
1. Potassium
(2-chloro-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis(hydroxym-
ethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
represented by the following formula (1): ##STR00007##
2. Potassium
(2-bromo-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis(hydroxyme-
thyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
represented by the following formula (2): ##STR00008##
3. A crystal of the compound of claim 1.
4. (canceled)
5. The crystal of claim 3, which shows characteristic peaks at
diffraction angles 2.theta. of 3.82.+-.2.degree.,
7.64.+-.2.degree., 11.48.+-.2.degree., 19.06.+-.2.degree.,
23.08.+-.2.degree., 25.22.+-.2.degree., and 26.98.+-.2.degree. in
powder X-ray diffraction obtained by irradiation with copper
K.alpha. radiation.
6. A crystal of the compound of claim 2.
7. (canceled)
8. The crystal of claim 6, which shows characteristic peaks at
diffraction angles 2.theta. of 7.66.+-.2.degree.,
15.36.+-.2.degree., 19.08.+-.2.degree., 23.76.+-.2.degree.,
25.26.+-.2.degree., and 27.04.+-.2.degree. in powder X-ray
diffraction obtained by irradiation with copper K.alpha.
radiation.
9. A pharmaceutical composition, comprising the compound of claim 1
and one or more pharmaceutical additives.
10. A method of suppressing the production of an inflammatory
mediator caused by intracellular signal transduction or cell
activation induced by endotoxin, comprising administering the
pharmaceutical composition of claim 9 to a warm-blooded animal or
human.
11. A method of treating sepsis, comprising administering the
compound of claim 1 to a warm-blooded animal or human.
12. A pharmaceutical composition, comprising the crystal of claim 3
and one or more pharmaceutical additives.
13. A method for producing the compound of claim 1, comprising
adding a solution of potassium 2-ethylhexanoate hydrate in ethyl
acetate dropwise to a solution or suspension of
ethyl(2R,3R,8R)-8-[N-(2-chloro-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxym-
ethyl)-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate.
14. A method for producing the compound of claim 2, comprising
adding
ethyl(2R,3R,8R)-8-[N-(2-bromo-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxyme-
thyl)-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate dropwise to a
solution of potassium 2-ethylhexanoate hydrate in ethyl
acetate.
15. A method of suppressing the production of an inflammatory
mediator caused by intracellular signal transduction or cell
activation induced by endotoxin, comprising administering the
compound of claim 1 to a warm-blooded animal or human.
16. A method of treating sepsis, comprising administering the
pharmaceutical composition of claim 9 to a warm-blooded animal or
human.
17. A pharmaceutical composition, comprising the compound of claim
2 and one or more pharmaceutical additives.
18. A method of suppressing the production of an inflammatory
mediator caused by intracellular signal transduction or cell
activation induced by endotoxin, comprising administering the
compound of claim 2 to a warm-blooded animal or human.
19. A method of suppressing the production of an inflammatory
mediator caused by intracellular signal transduction or cell
activation induced by endotoxin, comprising administering the
pharmaceutical composition of claim 17 to a warm-blooded animal or
human.
20. A method of treating sepsis, comprising administering the
compound of claim 2 to a warm-blooded animal or human.
21. A method of treating sepsis, comprising administering the
pharmaceutical composition of claim 17 to a warm-blooded animal or
human.
22. A pharmaceutical composition, comprising the crystal of claim 6
and one or more pharmaceutical additives.
Description
TECHNICAL FIELD
[0001] The present invention relates to crystals of substituted
cycloalkene derivatives, which have activity to suppress
intracellular signal transduction or cell activation in various
cells such as monocytes, macrophages, and vascular endothelial
cells induced by endotoxin, and to suppress the production of an
inflammatory mediator such as TNF-.alpha. caused by the
intracellular signal transduction or cell activation, and which are
useful as a prophylactic and/or therapeutic agent for various
diseases such as sepsis (septic shock, disseminated intravascular
coagulation, multiple organ failure, etc.).
BACKGROUND ART
[0002] Endotoxin (lipopolysaccharide: LPS), which is a membrane
component of bacteria, acts on cells such as monocytes,
macrophages, and vascular endothelial cells, induces the
overproduction of various inflammatory mediators such as
TNF-.alpha. and the like, causes sudden hypotension, blood
coagulation disorders, circulatory disturbances, and the like in
addition to systemic inflammatory responses, and thus develops
sepsis (see, for example, Non-patent document 2).
Lipopolysaccharide and Lipid A, which corresponds to a partial
structure thereof, activate intracellular signal transduction via
TLR4 (Toll-like receptor 4), which is a functional cell surface
receptor, after binding with CD14 (see, for example, Non-patent
document 3), whereby various cell responses represented by the
production of inflammatory mediators are initiated. Therefore, it
is considered that a substance capable of suppressing the
intracellular signal transduction or cell activation induced by
endotoxin, and various cell responses, which are induced by such
intracellular signal transduction or cell activation and
represented by the overproduction of inflammatory mediators such as
TNF-.alpha., can be effective prophylactic and therapeutic agents
for sepsis (see, for example, Non-patent documents 3 and 4, and
Patent documents 1 and 2).
[0003] Intracellular signal transduction or cell activation induced
by endotoxin, and various cell responses such as the overproduction
of inflammatory mediators including TNF-.alpha., etc. induced by
the intracellular signal transduction or cell activation lead to
development and progress of various diseases such as ischemic brain
disorder, arteriosclerosis, poor prognosis after coronary
angioplasty, heart failure, diabetes, diabetic complications,
arthritis, osteoporosis, osteopenia, autoimmune diseases, tissue
disorders and rejection after organ transplantation, bacterial
infection, viral infection, gastritis, pancreatitis, nephritis,
pneumonia, hepatitis, and leukemia, in addition to the
above-described sepsis (for example, Non-patent document 5 and
Patent document 3).
[0004] The present inventors made intensive studies in order to
solve the above problems, and as a result, they found that there
are a group of compounds having a desired effect among substituted
cycloalkene derivatives (Patent document 4). However, substituted
cycloalkene derivatives (in the free form) are amorphous and have
hygroscopicity. In view of this, the present inventors studied
pharmacologically acceptable salts thereof. However, it was not
easy to find crystals having storage and handling stability, and
therefore industrialization thereof was extremely difficult.
PRIOR ART DOCUMENTS
Patent Documents
[0005] Patent Document 1: JP-A-2000-178246
[0006] Patent Document 2: JP-A-2004-2370
[0007] Patent Document 3: WO 2000/41698
[0008] Patent Document 4: WO 2007/032362
Non-Patent Documents
[0009] Non-patent Document 1: Iqbal et al., Expert Opin. Emerging
Drugs, Vol. 7, page 111, 2002
[0010] Non-patent Document 2: Hawkins et al., Current Topics in
Medicinal Chemistry, Vol. 4, page 1147, 2004
[0011] Non-patent Document 3: Beutler, Nature, Vol. 430, pages
257-263, 2004
[0012] Non-patent Document 4: Kakutani et al., Inflammation
Research, Vol. 48, page 461, 1999
[0013] Non-patent Document 5: Donald N. Cook et al., Nature
Immunology, Vol. 5, pages 975-979, 2004
SUMMARY OF THE INVENTION
Problems to be Solved by the Invention
[0014] An object of the present invention is to provide crystals of
substituted cycloalkene derivatives having excellent storage and
handling stability.
Means for Solving the Problems
[0015] That is, the present invention is directed to:
[0016] (1)
potassium(2-chloro-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbony-
l)-2,3-bis(hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azani-
de represented by the following formula (1):
##STR00001##
[0017] (2)
potassium(2-bromo-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl-
)-2,3-bis(hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanid-
e represented by the following formula (2):
##STR00002##
[0018] (3) a crystal of the compound according to the above
(1);
[0019] (4) the crystal according to the above (3), which has the
X-ray diffraction pattern shown in FIG. 1 in powder X-ray
diffraction obtained by irradiation with copper K.alpha.
radiation;
[0020] (5) the crystal according to the above (3), which shows
characteristic peaks at diffraction angles 2.zeta. (.degree.) of
3.82, 7.64, 11.48, 19.06, 23.08, 25.22, and 26.98 (.+-.2,
respectively) in powder X-ray diffraction obtained by irradiation
with copper K.alpha. radiation;
[0021] (6) a crystal of the compound according to the above
(2);
[0022] (7) the crystal according to the above (6), which has the
X-ray diffraction pattern shown in FIG. 2 in powder X-ray
diffraction obtained by irradiation with copper K.alpha.
radiation;
[0023] (8) the crystal according to the above (6), which shows
characteristic peaks at diffraction angles 2.theta. (.degree.) of
7.66, 15.36, 19.08, 23.76, 25.26, and 27.04 (.+-.2, respectively)
in powder X-ray diffraction obtained by irradiation with a copper
K.alpha. radiation;
[0024] (9) a pharmaceutical composition, comprising the compound
according to the above (1) or (2) as an active ingredient;
[0025] (10) a pharmaceutical composition, comprising the compound
according to the above (1) or (2) as an active ingredient, for use
in suppressing the production of an inflammatory mediator caused by
intracellular signal transduction or cell activation induced by
endotoxin;
[0026] (11) a prophylactic and/or therapeutic agent for sepsis,
comprising the compound according to the above (1) or (2) as an
active ingredient;
[0027] (12) a pharmaceutical composition, comprising the crystal
according to any one of the about (1) to (8);
[0028] (13) a method for producing the compound according to the
above (1), characterized by adding a solution of potassium
2-ethylhexanoate hydrate in ethyl acetate dropwise to a solution or
suspension of
ethyl(2R,3R,8R)-8-[N-(2-chloro-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxym-
ethyl)-1,4- dioxaspiro[4.5]deca-6-ene-7-carboxylate; or
[0029] (14) a method for producing the compound according to the
above (2), characterized by adding
ethyl(2R,3R,8R)-8-[N-(2-bromo-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxyme-
thyl)-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate dropwise to a
solution of potassium 2-ethylhexanoate hydrate in ethyl
acetate.
Advantage of the Invention
[0030] According to the present invention, crystals of substituted
cycloalkene derivatives having excellent storage and handling
stability can be provided. Potassium salts (crystals) of
substituted cycloalkene derivatives according to the present
invention suppress the production of an inflammatory mediator
caused by intracellular signal transduction or cell activation
induced by endotoxin and are effective as a prophylactic and/or
therapeutic agent for sepsis.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] FIG. 1 shows a powder X-ray diffraction pattern of a crystal
obtained in Example 1. In the drawing, the ordinate indicates
diffraction intensity in units of counts/sec (cps), and the
abscissa indicates the value of diffraction angle 2.theta..
[0032] FIG. 2 shows a powder X-ray diffraction pattern of a crystal
obtained in Example 2. In the drawing, the ordinate indicates
diffraction intensity in units of counts/sec (cps), and the
abscissa indicates the value of diffraction angle 2.theta..
[0033] FIG. 3 shows moisture absorption and desorption equilibrium
curves of a compound (in the free form) of Comparative Example in a
moisture absorption test. In the drawing, the ordinate indicates
the weight of a sample, and the abscissa indicates the measurement
humidity. "Cycle 1 Sorp" represents "a moisture absorption
equilibrium curve (in the first cycle)", and change in weight
against relative humidity at the time of obtaining a constant
weight when the humidity was increased is plotted. "Cycle 1 Desorp"
represents "a moisture desorption equilibrium curve (in the first
cycle)", and change in weight against relative humidity at the time
of obtaining a constant weight when the humidity was decreased is
plotted.
[0034] FIG. 4 shows moisture absorption and desorption equilibrium
curves of a compound of Example 1 in a moisture absorpotion test.
In the drawing, the ordinate indicates the change in weight of a
sample, and the abscissa indicates the set measurement humidity.
"Cycle 1 Sorp" represents "a moisture absorption equilibrium curve
(in the first cycle)", and change in weight against relative
humidity at the time of obtaining a constant weight when the
humidity was increased is plotted. "Cycle 1 Desorp" represents "a
moisture desorption equilibrium curve (in the first cycle)", and
change in weight against relative humidity at the time of obtaining
a constant weight when the humidity was decreased is plotted.
[0035] FIG. 5 shows moisture absorption and desorption equilibrium
curves of a compound of Example 2 in a moisture absorption test. In
the drawing, the ordinate indicates the change in weight of a
sample, and the abscissa indicates the set measurement humidity.
"Cycle 1 Sorp" represents "a moisture absorption equilibrium curve
(in the first cycle)", and change in weight against relative
humidity at the time of obtaining a constant weight when the
humidity was increased is plotted. "Cycle 1 Desorp" represents "a
moisture desorption equilibrium curve (in the first cycle)", and
change in weight against relative humidity at the time of obtaining
a constant weight when the humidity was decreased is plotted.
MODE FOR CARRYING OUT THE INVENTION
[0036] The present invention relates to a crystal of
potassium(2-chloro-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis-
(hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
represented by the following formula (1) (hereinafter sometimes
referred to as Compound (1) in this description):
##STR00003##
or
potassium(2-bromo-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-b-
is(hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
represented by the following formula (2) (hereinafter sometimes
referred to as Compound (2) in this description).
##STR00004##
[0037] Here, a crystal refers to a solid whose internal structure
is three-dimensionally composed of a regular repetition of
constituent atoms (or a group thereof), and is distinguished from
an amorphous solid which does not have such a regular internal
structure. Whether or not a solid is a crystal can be examined by a
known crystallographic method (such as powder X-ray diffraction
measurement or differential scanning calorimetric analysis). For
example, a solid is subjected to powder X-ray diffraction
measurement using an X-ray obtained by irradiation with copper
K.alpha. radiation. The solid is determined to be a crystal when
distinct peaks are observed in the X-ray diffraction pattern, while
the solid is determined to be amorphous when no distinct peaks are
observed. The solid is determined to be a crystal whose
crystallinity is low when the peaks can be read but are not
distinct (for example, broad), and such a crystal whose
crystallinity is low is also encompassed within the crystal of the
present invention.
[0038] Even the same compound can form crystals having a plurality
of different internal structures and physicochemical properties
(crystal polymorphism) depending on the conditions for
crystallization. The crystal of the present invention may be any of
these crystal polymorphic forms and may be a mixture of two or more
crystal polymorphic forms.
[0039] The crystal of the present invention may sometimes form a
hydrate by allowing the crystal to stand in the air to absorb
water, thereby having attached water, or by heating the crystal to
25 to 150.degree. C. in usual atmospheric conditions, etc. Further,
the crystal of the present invention may sometimes contain a
solvent used at the time of crystallization as an attached residual
solvent or a solvate.
[0040] In this description, the crystal of the present invention is
sometimes represented on the basis of powder X-ray diffraction
data. In powder X-ray diffraction, the measurement and analysis may
be performed by methods conventionally used in this field, and for
example, the powder X-ray diffraction can be performed by the
method described in the Examples. Further, in general, in the case
of a hydrated or dehydrated crystal, the lattice constant thereof
is changed by the addition or removal of water of crystallization,
and therefore the diffraction angle (2.theta.) in powder X-ray
diffraction may sometimes be changed. Further, the peak intensity
may sometimes be changed due to a difference in a crystal growth
surface or the like (crystal habit), etc. Therefore, when the
crystal of the present invention is represented on the basis of
powder X-ray diffraction data, a crystal having an identical peak
diffraction angle and X-ray diffraction pattern in powder X-ray
diffraction, and also hydrated and dehydrated crystals obtained
from the crystal, are encompassed within the scope of the present
invention.
[0041] In powder diffraction measurement using copper K.alpha.
radiation, generally, a sample is irradiated with copper K.alpha.
radiation (in which K.alpha.1 and K.alpha.2 radiations are not
separated). An X-ray diffraction pattern can be obtained by
analyzing diffraction derived from K.alpha. radiation, and can also
be obtained by analyzing only diffraction derived from K.alpha.1
radiation taken from diffraction derived from K.alpha. radiation.
In the present invention, the powder X-ray diffraction pattern
obtained by irradiation with K.alpha. radiation includes an X-ray
diffraction pattern obtained by analyzing diffraction peaks derived
from K.alpha. radiation and an X-ray diffraction pattern obtained
by analyzing diffraction derived from K.alpha.1 radiation, and is
preferably an X-ray diffraction pattern obtained by analyzing
diffraction derived from K.alpha.1 radiation.
[0042] One preferred embodiment of the crystal of the present
invention is a crystal of
potassium(2-chloro-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis-
(hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
referred to as Compound (1). The crystal of Compound (1) has the
X-ray diffraction pattern shown in FIG. 1 in a powder diffraction
pattern obtained by irradiation with copper K.alpha. radiation.
Further, the crystal of Compound (1) has characteristic peaks at
diffraction angles 2.theta. (.degree.) of 3.82, 7.64, 11.48, 19.06,
23.08, 25.22, and 26.98. Here, the "characteristic peak" refers to
a peak having a relative intensity of 9 or more when the maximum
peak intensity in the powder X-ray diffraction is taken as 100.
[0043] Another preferred embodiment of the crystal of the present
invention is a crystal of
potassium(2-bromo-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis(-
hydroxymethyl)-1,4-dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
referred to as a crystal of Compound (2). The crystal of Compound
(2) has the X-ray diffraction pattern shown in FIG. 2 in a powder
diffraction pattern obtained by irradiation with copper K.alpha.
radiation. Further, the crystal of Compound (2) has characteristic
peaks at diffraction angles 2.theta. (.degree.) of 7.66, 15.36,
19.08, 23.76, 25.26, and 27.04. Here, the "characteristic peak"
refers to a peak having a relative intensity of 10 or more when the
maximum peak intensity in the powder X-ray diffraction is taken as
100.
[0044] In the powder X-ray diffraction patterns shown in FIGS. 1
and 2, the ordinate indicates diffraction intensity [counts/sec
(cps)], and the abscissa indicates the diffraction angle 2.theta.
(.degree.). The 2.theta. can be slightly changed in its position
and relative intensity depending on the measurement conditions and
the like, therefore even when 2.theta. is slightly changed, the
identification of the crystal form should be appropriately
determined by reference to the pattern of the entire spectrum. The
limit of such error is generally within the range of .+-.2,
preferably within the range of .+-.1, more preferably within the
range of .+-.0.5, further more preferably within the range of
.+-.0.2.
[0045] Further, as is well known in the field of crystallography,
also the intensities of the respective diffraction peaks can be
changed by various factors (including preferred orientation
occurring in a specific crystalline form and the effect of grain
size), and therefore, the relative intensities of the
above-described main peaks for identifying the crystal of the
present invention can also be changed, and these crystals are also
encompassed within the crystal of the present invention.
[0046] The Compound (1) or (2) of the present invention has
activity to suppress the production of an inflammatory mediator
caused by intracellular signal transduction or cell activation
induced by endotoxin, and also has excellent storage and handling
stability, and therefore is useful as a pharmaceutical. Further
such a pharmaceutical is preferably used for warm-blooded animals,
more preferably used for humans.
[0047] In the case where Compound (1) and Compound (2) of the
present invention are used as a therapeutic agent or a prophylactic
agent for the above-described diseases, the compound per se or as a
mixture with an appropriate pharmacologically acceptable excipient,
diluent, or the like can be administered orally in the form of a
tablet, a capsule, a granule, a powder, a syrup, or the like, or
parenterally in the form of an injection, a suppository, or the
like.
[0048] These pharmaceutical preparations are prepared in accordance
with known methods using additives such as an excipient (for
example, an organic excipient such as a sugar derivative such as
lactose, sucrose, glucose, mannitol, or sorbitol; a starch
derivative such as corn starch, potato starch, pregelatinized
starch or dextrin; a cellulose derivative such as crystalline
cellulose; gum arabic; dextrane; or pullulan; or an inorganic
excipient such as a silicate derivative such as light silicic
anhydride, synthetic aluminum silicate, calcium silicate, or
magnesium aluminometasilicate; a phosphate such as calcium hydrogen
phosphate; a carbonate such as calcium carbonate; or a sulfate such
as calcium sulfate can be used), a lubricant (for example, stearic
acid, a metal salt of stearic acid such as calcium stearate or
magnesium stearate; talc; a wax such as beeswax or spermaceti wax;
boric acid; adipic acid; a sulfate such as sodium sulfate; glycol;
fumaric acid; sodium benzoate; D- or L-leucine; a lauryl sulfate
such as sodium lauryl sulfate or magnesium lauryl sulfate; a
silicic acid such as silicic anhydride or silicate hydrate; or any
of the above-described starch derivatives can be used), a binder
(for example, hydroxypropyl cellulose, hydroxypropyl methyl
cellulose, polyvinylpyrrolidone, macrogol, or a compound similar to
any of the above-described excipients can be used), a disintegrant
(for example, a cellulose derivative such as low-substituted
hydroxypropyl cellulose, carboxymethyl cellulose, calcium
carboxymethyl cellulose, or internally crosslinked sodium
carboxymethyl cellulose; a chemically modified water-soluble
polymer such as carboxymethyl starch, sodium carboxymethyl starch,
or crosslinked polyvinylpyrrolidone; or any of the above-described
starch derivatives can be used), an emulsifier (for example, a
colloidal clay such as bentonite or veegum; a metal hydroxide such
as magnesium hydroxide or aluminum hydroxide; an anionic surfactant
such as sodium lauryl sulfate or calcium stearate; a cationic
surfactant such as benzalkonium chloride; or a nonionic surfactant
such as a polyoxyethylene alkyl ether, a polyoxyethylene sorbitan
fatty acid ester, or a sucrose fatty acid ester can be used), a
stabilizer (for example, a paraoxybenzoic acid ester such as methyl
paraben or propyl paraben; an alcohol such as chlorobutanol, benzyl
alcohol, or phenyl ethyl alcohol; benzalkonium chloride; a phenol
such as phenol or cresol; thimerosal; dehydroacetic acid; or sorbic
acid can be used) and a corrigent (for example, a commonly used
sweetener, acidifier, flavor, or the like can be used) or a
diluent.
EXAMPLES
[0049] Hereinafter, the present invention will be described in
further detail with reference to Examples and Test Examples.
Example 1
Potassium(2-chloro-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis(-
hydroxymethyl)-1,4-
dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
##STR00005##
[0051] 100 mg (0.208 mmol) of ethyl
(2R,3R,8R)-8-[N-(2-chloro-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxymethyl-
)-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate [a compound described
as a low polarity compound (first peak) of Example 162 in WO
2007/032362] was dissolved in 1 mL of ethyl acetate, and 2 mL of a
solution containing 38 mg (0.208 mmol) of potassium
2-ethylhexanoate hydrate in ethyl acetate was added thereto under
stirring at room temperature. The resulting reaction solution was
concentrated under reduced pressure, and to the residue, diethyl
ether was added. A precipitated solid was obtained by filtration
and washed with diethyl ether, whereby 92 mg of a crude product was
obtained as an amorphous solid. Then, the crude product was
dissolved in ethyl acetate, and the resulting solution was heated
under reflux at 130.degree. C. for 2 hours. After the resulting
mixture was cooled, a precipitated solid was obtained by filtration
and washed with ethyl acetate, whereby the title compound was
obtained as a white crystal.
[0052] .sup.1H-NMR spectrum (400 MHz, DMSO-d.sub.6) .delta. ppm:
7.40 (1H, dd, J=9 Hz, 6 Hz), 7.07-6.97 (1H, m), 6.84-6.75 (1H, m),
6.46 (1H, s),
[0053] 4.98 (1H, t, J=6 Hz), 4.82 (1H, t, J=6 Hz), 4.11-4.02 (1H,
m), 4.02-3.93 (1H, m),
[0054] 3.93-3.80 (3H, m), 3.63-3.41 (4H, m), 2.78-2.65 (1H, m),
2.34-2.25 (1H, m),
[0055] 1.87-1.74 (1H, m), 1.65-1.58 (1H, m), 1.14 (3H, t, J=7
Hz)
[0056] The powder X-ray diffraction pattern is shown in FIG. 1.
<Another Method>
[0057] 300 mg (0.625 mmol) of
ethyl(2R,3R,8R)-8-[N-(2-chloro-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxym-
ethyl)-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate was dissolved in
2 mL of ethyl acetate, and 4 mL of a solution containing 114 mg
(0.625 mmol) of potassium 2-ethylhexanoate hydrate in ethyl acetate
was added thereto under stirring at room temperature. The resulting
reaction solution was concentrated under reduced pressure, and to
the residue, diethyl ether was added. A precipitated solid was
obtained by filtration and washed with diethyl ether, whereby a
crude product was obtained as an amorphous solid. Then, the crude
product was dissolved in 3 mL of ethyl acetate, and a seed crystal
obtained by the above-described method was added thereto, and the
resulting mixture was heated under reflux at 100.degree. C. for 5
minutes. After the resulting mixture was cooled, a precipitated
solid was obtained by filtration and washed with ethyl acetate,
whereby 236 mg of the title compound was obtained as a white
crystal (yield: 73%).
Example 2
Potassium(2-bromo-4-fluorophenyl){[(2R,3R,8R)-7-(ethoxycarbonyl)-2,3-bis(h-
ydroxymethyl)-1,4-
dioxaspiro[4.5]deca-6-en-8-yl]sulfonyl}azanide
##STR00006##
[0059] 422 mg (2.31 mmol) of potassium 2-ethylhexanoate hydrate was
dissolved in 23 mL of ethyl acetate, and 1.215 g (2.31 mmol) of
ethyl(2R,3R,8R)-8-[N-(2-bromo-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxyme-
thyl)-1,4- dioxaspiro[4.5]deca-6-ene-7-carboxylate [a compound
described as a low polarity compound (first peak) of Example 166 in
WO 2007/032362] was added thereto under stirring at room
temperature. The resulting reaction solution was concentrated under
reduced pressure, and to the residue, diethyl ether was added. A
precipitated solid was obtained by filtration and washed with
diethyl ether, whereby 954 mg of a crude product was obtained as an
amorphous solid. Then, the crude product was dissolved in 6 mL of
ethyl acetate, and the resulting solution was heated under stirring
at 70.degree. C. for 5 minutes. After the resulting mixture was
cooled, a precipitated solid was obtained by filtration and washed
with ethyl acetate and diethyl ether, whereby 850 mg of the title
compound was obtained as a white crystal (yield: 65%).
[0060] .sup.1H-NMR spectrum (400 MHz, DMSO-d.sub.6) .delta.
ppm:
[0061] 7.39 (1H, dd, J=9 Hz, 6 Hz), 7.17 (1H, dd, J=9 Hz, 3 Hz),
6.83 (1H, dt, J=9 Hz, 3 Hz), 6.45 (1H, s), 4.98 (1H, t, J=5 Hz),
4.83 (1H, t, J=5 Hz), 4.13-3.81 (5H, m), 3.64 -3.41 (4H, m),
2.84-2.63 (1H, m), 2.35-2.26 (1H, m), 1.85-1.73 (1H, m), 1.65-1.56
(1H, m), 1.14 (3H, t, J=7 Hz)
[0062] The powder X-ray diffraction pattern is shown in FIG. 2.
<Another Method>
[0063] 102 mg (0.560 mmol) of potassium 2-ethylhexanoate hydrate
was dissolved in 5 mL of ethyl acetate, and 280 mg (0.534 mmol) of
ethyl
(2R,3R,8R)-8-[N-(2-bromo-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxymethyl)-
-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate was added thereto
under stirring at room temperature. The resulting reaction solution
was concentrated under reduced pressure, and to the residue,
diethyl ether was added. A precipitated solid was obtained by
filtration and washed with diethyl ether, whereby a crude product
was obtained as an amorphous solid. Then, the crude product was
dissolved in 1 mL of ethanol, and the resulting solution was heated
under stirring at 70.degree. C. for 1 minute. After the resulting
mixture was cooled, a precipitated solid was obtained by filtration
and washed with ethanol and diethyl ether, whereby 197 mg of the
title compound was obtained as a white crystal (yield: 66%).
Example 3
Measurement of Powder X-Ray Diffraction
[0064] Rint TTR-III (provided with a scintillation counter tube and
a long slit for removing K.sub..beta. radiations) manufactured by
Rigaku Corporation was used. A sample was placed uniformly on a
non-reflective sample holder, and measurement was performed under
the following conditions.
<Analytical Conditions>
[0065] X-ray species: Cu--K.sub.a (wavelength: 1.54 A), Tube
voltage: 50 kV, Tube current: 300 mA, Scanning rate: 2.degree./min,
Step: 0.02.degree., Scanning range (2q) : 2.degree. to 60.degree.,
Divergence slit: 0.5 mm, Scattering slit: 0.5 mm, Receiving slit:
0.5 mm
<Measurement Results>
[0066] The powder X-ray diffraction pattern obtained by measuring
the crystal of the compound obtained in Example 1 in accordance
with the above-described method is shown in FIG. 1. Peaks having a
relative intensity of 9 or more when the maximum peak intensity in
FIG. 1 is taken as 100 are shown in Table 1.
TABLE-US-00001 TABLE 1 Relative 2.theta. d value intensity 3.82
23.11 24 7.64 11.56 100 11.48 7.70 9 19.06 4.65 12 23.08 3.85 12
25.22 3.53 9 26.98 3.30 30
[0067] The powder X-ray diffraction pattern obtained by measuring
the crystal of the compound obtained in Example 2 in accordance
with the above-described method is shown in FIG. 2. Peaks having a
relative intensity of 10 or more when the maximum peak intensity in
FIG. 2 is taken as 100 are shown in Table 2.
TABLE-US-00002 TABLE 2 Relative 2.theta. d value intensity 7.66
11.53 100 15.36 5.76 12 19.08 4.65 11 23.76 3.74 13 25.26 3.52 13
27.04 3.29 28
Example 4
Moisture Absorption Test
[0068] A sample was weighed in a glass sample cup, and the weight
thereof was measured under the following conditions.
<Measurement Conditions>
[0069] Measuring device: DVS Advantage, manufactured by Surface
Measurement System Ltd.
[0070] Measurement humidity: 10, 20, 30, 40, 50, 60, 65, 70, 75,
80, 85, 90, 95, 95, 90, 85, 80, 75, 70, 65, 60, 50, 40, 30, 25, 20,
and 10% RH
[0071] Measurement temperature: 25.degree. C., Minimum exposure
time: 15 min, Maximum exposure time: 120 min, Step transition
condition: within 0.006 wt. %
[0072] The appearance of the sample at the time of completion of
the measurement was confirmed.
<Measurement Results>
[0073] As shown in Table 4 and FIGS. 3, 4, and 5,
ethyl(2R,3R,8R)-8-[N-(2-chloro-4-fluorophenyl)sulfamoyl]-2,3-bis(hydroxym-
ethyl)-1,4-dioxaspiro[4.5]deca-6-ene-7-carboxylate (a compound in
the free form described as a low polarity compound (first peak) of
Example 162 in WO 2007/02362) used as the compound of Comparative
Example showed high hygroscopicity at every measurement humidity
(see FIG. 3), however, the compounds of Examples 1 and 2 did not
show hygroscopicity (see FIGS. 4 and 5). In particular, the
compound of Example 1 did not show hygroscopicity at any
measurement humidity in a consistent manner (see FIG. 4).
Example 5
Solubility Test
<Test Method>
[0074] 150 mg of each of the compounds of Examples 1 and 2 and
Comparative Example 1 was dissolved in 5 mL of an aqueous phosphate
solution containing sodium chloride (containing 150 mM NaCl and 10
mM NaH.sub.2PO.sub.3.2H.sub.2O). After the resulting mixture was
stirred for 30 minutes while measuring the pH of the mixture, a 0.5
mL portion of the resulting suspension was taken and filtered
through a syringe filter. A 100 gL portion of the obtained filtrate
was transferred into a 10-mL volumetric flask and diluted to 10 mL
with an aqueous solution of 50% MeCN, whereby a sample solution was
prepared. To the suspension, concentrated HCl was added dropwise to
adjust the pH thereof to around 7.5.
[0075] Measurement was performed using HPLC under the following
conditions.
<Analytical Method Conditions>
[0076] HPLC system: Water's Alliance
[0077] Column: XTerra MS C18 3.5 .mu.m, Column Size: 4.6.times.100
mm
[0078] Column Temp: 40.degree. C., Flow Rate: 1.2 mL/min
[0079] Solvent A: 0.1% phosphoric acid in water, Solvent B: 0.1%
phosphoric acid in MeCN
Gradient Schedule
TABLE-US-00003 [0080] TABLE 3 Time Total flow % A % B Curve 0 1.2
95 5 15 1.2 5 95 6 15.01 1.2 95 5 11 20 1.2 95 5 6
<Measurement Results>
[0081] As shown in Table 4, the compound (in the free form) of
Comparative Example had low solubility. Meanwhile, the compounds of
Examples 1 and 2 showed a solubility of 13.8 mg/mL and 9.6 mg/mL,
respectively.
Example 6
Solution Stability Test
<Test Method>
[0082] Each of the compounds (Examples 1 and 2 and Comparative
Example) of the present invention was precisely weighed and placed
in a 10-mL volumetric flask and dissolved in PBS at pH 6. Then, 1 N
HCl was added thereto to adjust the solution of the compound of
Example 1 to pH 6, the solution of the compound of Example 2 to pH
7, and the solution of the compound of Comparative Example to pH 8.
The above flasks as such and the flasks covered with aluminum foil
for light shielding as controls were placed in a light tester at
25.degree. C. After one day, each solution was diluted to 10 mL
with 50% MeCN. The concentration of each solution was calculated by
performing HPLC under the same conditions as used in the
above-described solubility test, and the change in concentration
from the control was calculated.
<Measurement Results>
[0083] As shown in Table 4, the values for the solution stability
after storage for 1 day were 99.4% in the case of the compound (in
the free form) of Comparative Example, 99.9% in the case of the
compound of Example 1, and 99.4% in the case of the compound of
Example 2.
Example 7
Evaluation for Chemical Stability
<Test Method>
[0084] In a low-humidity room (25.degree. C., 30% RH), a sample was
precisely weighed in a quartz sample cup and placed in a 10-mL
volumetric flask. Then, the flask was left in a desiccator
containing silica gel for 3 days while keeping the mouth of the
flask open. Thereafter, the flask was further stored in an
environment at a temperature of 25.degree. C. and a humidity of 30%
for 4 weeks while keeping the mouth of the flask open. Then, the
residual amount was measured by performing HPLC under the same
conditions as used in the solubility test in Example 5.
<Measurement Results>
[0085] As shown in Table 4, it was impossible to perform this test
for the compound (in the free form) of Comparative Example because
the compound is hygroscopic, however, the compounds of Examples 1
and 2 showed a residual amount of 98.9% and 98.8%, respectively,
which revealed that the compounds of Examples 1 and 2 are
stable.
TABLE-US-00004 TABLE 4 Comparative Example Example 1 Example 2 Free
form or Free form Potassium salt Potassium salt potassium salt
Crystalline Amorphous Crystalline Crystalline form Hygroscopicity
With hygroscopicity Without Without 2.6% (relative hygroscopicity
hygroscopicity humidity 10.fwdarw.80%) <0.1% (relative <0.1%
(relative Changed into candy- humidity humidity like form
10.fwdarw.80%) 10.fwdarw.80%) no change in no change in appearance
appearance Solubility Since the compound 13.8 mg/ml 9.6 mg/ml (PBS
pH 7.5) is transformed into a viscous solid in an aqueous solution,
it was impossible to measure the concentration in a reproducible
manner. Solution 99.4% 99.9% 99.4% stability McIlvaine buffer pH 8
PBS pH 6 PBS pH 7 (25.degree. C., 1 day) Chemical Due to high 98.9%
(relative 98.8% (relative stability hygroscopicity, it humidity
30%) humidity 30%) (25.degree. C., was impossible to 4 weeks)
perform this test.
Test Example 1
Suppressive Effect on Production of TNF-.alpha. in Cells Stimulated
with Endotoxin (In Vitro)
[0086] The rate of suppression of TNF-.alpha. production when human
monocytic cell line U937 was stimulated with endotoxin was
determined for the compound of the present invention. Specifically,
to RPMI-1640 medium containing 10% (volume %) of inactivated
newborn bovine serum, 12-O-tetradecanoylphorbol-13-acetate was
added to give a final concentration of 30 ng/ml. U937 cells were
suspended in the medium and plated into a 96-well culture plate
(Sumilon) so that the number of cells per well/volume was
2.times.10.sup.4/0.1 mL, and the culture plate was then incubated
for 3 days at 37.degree. C. in a carbon dioxide incubator with 5%
CO.sub.2 and 100% humidity. After completion of the incubation, the
culture supernatant was removed. The compound of the present
invention was added to each well at different concentrations, and
also lipopolysaccharide (LPS) (E. coli 0111:B4, Sigma) was added
thereto to give a final concentration of 30 ng/ml. After the
culture plate was incubated in the carbon dioxide incubator again
for 4.5 hours, the culture supernatant was collected. By using a
384-well half area black plate (Greiner) and an HTRF quantitative
determination kit manufactured by Cisbio International, the
concentration of TNF-.alpha. in the culture supernatant was
measured as time-resolved fluorescence by Discovery (Packard). From
the measured value in the absence of LPS (X), the measured value in
the absence of the compound of the present invention (Y), and the
measured value in the presence of the compound of the present
invention (Z), the rate of suppression of TNF-.alpha. production
was obtained according to the following calculation formula
[I].
[0087] Rate of suppression of TNF-.alpha. production
(%)={1-(Z--X)/(Y--X)}.times.100 [I]
<Suppressive Effect on Production of TNF-.alpha. (In
Vitro)>
TABLE-US-00005 [0088] TABLE 5 <Suppressive Effect on Production
of TNF-.alpha. (in vitro)> Rate of suppression of TNF-.alpha.
production [%] (concentration of Test compound test compound: 100
nM) Compound of 89 Example 1
[0089] As shown in Table 5, in this test, the compound of the
present invention showed an excellent suppressive effect on the
production of TNF-.alpha. in cells stimulated with endotoxin.
Test Example 2
Suppressive Effect on Elevation of Blood TNF-.alpha. Level (In
Vivo)
[0090] The suppressive effect of the compound of the present
invention on the elevation of blood TNF-.alpha. level was studied.
A test for the elevation of blood TNF-.alpha. level was performed
in accordance with the method of Parant et al. described in Journal
of Leukocyte Biology, Vol. 47, page 164 (1990).
[0091] In the test, 3 to 4 male Sprague Dawley rats (8 to 9 weeks
of age) were used for each group.
[0092] At 4 hours before the administration of LPS, muramyl
dipeptide dissolved in physiological saline (1 mg/mL) was
administered to each rat through the tail vein at a dose of 1
mL/kg. At 0.5 hours before the administration of LPS, the rats were
anaesthetized with pentobarbital (40 mg/kg), and the compound of
the present invention dissolved in a 5% dimethyl acetamide/95%
polyethylene glycol 400 solution was administered to each rat
through the right femoral vein at a dose of 1 mL/kg. To the control
group, a 5% dimethyl acetamide/95% polyethylene glycol 400 solution
was administered at a dose of 1 mL/kg. LPS dissolved in
physiological saline (3 .mu.g/mL) was administered to each rat
through the left femoral vein at a dose of 1 mL/kg. At 2 hours
after the administration of LPS, the blood was collected using a
3.8% (w/v) sodium citrate solution as an anticoagulant, and the
blood plasma was separated by centrifugation (10,000 g, 5 minutes,
4.degree. C.). The plasma TNF-.alpha. level was determined using a
TNF-.alpha. quantitative determination kit (BioSource
International, Inc.). From the blood TNF-.alpha. level in the
control group (X) and the blood TNF-.alpha. level in the group
administered with the compound of the present invention (Y), the
rate of suppression of TNF-.alpha. production was calculated
according to the following calculation formula [II].
[0093] Rate of suppression of TNF-.alpha. production
(%)={1-Y/X}.times.100 [II]
<Suppressive Effect on Production of TNF-.alpha. (In
Vivo)>
TABLE-US-00006 [0094] TABLE 6 <Suppressive Effect on Production
of TNF-.alpha. (in vivo)> Rate of suppression of TNF-.alpha.
production [%] (concentration of Test compound test compound: 0.3
mg/kg) Compound of 96 Example 1 Compound of 81 Example 2
[0095] As shown in Table 6, in this test, the compounds of the
present invention showed an excellent suppressive effect on the
elevation of blood TNF-.alpha. level.
INDUSTRIAL APPLICABILITY
[0096] According to the present invention, crystals of substituted
cycloalkene derivatives having excellent storage and handling
stability can be provided. The potassium salts of substituted
cycloalkene derivatives of the present invention suppress the
production of an inflammatory mediator caused by intracellular
signal transduction or cell activation induced by endotoxin, and
therefore are effective as a prophylactic and/or therapeutic agent
for sepsis.
* * * * *