U.S. patent application number 15/848267 was filed with the patent office on 2018-08-23 for hydantoin derivative.
This patent application is currently assigned to Chugai Seiyaku Kabushiki Kaisha. The applicant listed for this patent is Chugai Seiyaku Kabushiki Kaisha. Invention is credited to Toru Esaki, Yoshikazu Nishimura, Tatsuya Tamura.
Application Number | 20180237436 15/848267 |
Document ID | / |
Family ID | 50934349 |
Filed Date | 2018-08-23 |
United States Patent
Application |
20180237436 |
Kind Code |
A1 |
Nishimura; Yoshikazu ; et
al. |
August 23, 2018 |
HYDANTOIN DERIVATIVE
Abstract
The present invention provides compounds represented by formula
(1) below and pharmacologically acceptable salts thereof:
##STR00001## wherein R.sub.1, R.sub.2, R.sub.3, and R.sub.4 are as
defined in the claims.
Inventors: |
Nishimura; Yoshikazu;
(Shizuoka, JP) ; Esaki; Toru; (Shizuoka, JP)
; Tamura; Tatsuya; (Shizuoka, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Chugai Seiyaku Kabushiki Kaisha |
Tokyo |
|
JP |
|
|
Assignee: |
Chugai Seiyaku Kabushiki
Kaisha
Tokyo
JP
|
Family ID: |
50934349 |
Appl. No.: |
15/848267 |
Filed: |
December 20, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15214729 |
Jul 20, 2016 |
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15848267 |
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14426408 |
Mar 6, 2015 |
9428505 |
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PCT/JP2013/083022 |
Dec 10, 2013 |
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15214729 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07D 519/00 20130101;
A61P 3/14 20180101; A61P 3/00 20180101; A61P 5/00 20180101; A61P
43/00 20180101; A61P 19/00 20180101; A61P 5/18 20180101; C07D
471/10 20130101 |
International
Class: |
C07D 471/10 20060101
C07D471/10; C07D 519/00 20060101 C07D519/00 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 10, 2012 |
JP |
2012-269178 |
Claims
1. A compound represented by general formula (1) below or a
pharmacologically acceptable salt, or a hydrate or a solvate
thereof: ##STR00058## (wherein, when R1 and R2 are not both
hydrogens, R1 and R2 are independently: 1) hydrogen; 2) halogen
atom; 3) an alkyl group comprising one or two carbons that may be
substituted with one to five fluorine atoms; or 4) an alkoxy group
comprising one or two carbons that may be substituted with one to
five fluorine atoms; or R1 and R2 bond with each other to form a
group represented by the formula below: ##STR00059## (wherein each
* indicates the position of bonding with the phenyl portion); and
R3 and R4 are independently a methyl group that may be substituted
with one to three fluorine atoms; or R3 and R4, together with a
bound carbon atom, form a three- to six-membered carbocyclic ring
(wherein, one of the carbon atoms forming the ring may be replaced
with an oxygen atom, a sulfur atom, or a methyl-substituted or
unsubstituted nitrogen atom).
2. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein R1 and R2 are selected
from the combinations below: 1) R1 is a hydrogen atom or a halogen
atom, and R2 is a hydrogen atom, a trifluoromethyl group, or a
trifluoromethoxy group (provided that R1 and R2 are not both
hydrogen atoms); 2) R1 is a trifluoromethyl group or a
trifluoromethoxy group, and R2 is a hydrogen atom or a halogen
atom; 3) R1 and R2 bond with each other to form a group represented
by the formula below: ##STR00060## (wherein, each * indicates the
position of bonding with the phenyl portion); and R3 and R4 are
methyl groups; or R3 and R4, together with a bound carbon atom,
form a ring selected from below: ##STR00061## (wherein * indicates
the position of bonding with the imidazolidine-2,4-dione
portion).
3. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein R1 and R2 are selected
from the combinations below: 1) R1 is a trifluoromethoxy group and
R2 is a fluorine atom; 2) R1 is a bromine atom and R2 is a hydrogen
atom; 3) R1 is a trifluoromethoxy group and R2 is a fluorine atom;
4) R1 is a fluorine atom and R2 is a trifluoromethoxy group; 5) R1
is a trifluoromethyl group and R2 is a hydrogen atom; 6) R1 is a
hydrogen atom and R2 is a trifluoromethoxy group; 7) R1 and R2 bond
with each other to form a group represented by the formula below:
##STR00062## (wherein each * indicates the position of bonding with
the phenyl portion); and R3 and R4 are methyl groups; or R3 and R4,
together with a bound carbon atom, form a ring selected from below:
##STR00063## (wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion).
4. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein R3 and R4 are methyl
groups.
5. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein R3 and R4, together with a
bound carbon atom, form a ring selected from below: ##STR00064##
(wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion).
6. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein the compound is selected
from the group consisting of:
1-(4-(2-((2-(4-fluoro-3-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro-
[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidaz-
olidine-2,4-dione;
1-(4-(2-((2-(3-bromophenyl)-4-oxo-1,3,8-triazaspiro[4.5]deca-1-en-8-yl)su-
lfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidazolidine-2,4-dione;
1-(4-(2-((2-(4-fluoro-3-(trifluoromethyl)phenyl)-4-oxo-1,3,8-triazaspiro[-
4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidazo-
lidine-2,4-dione;
1-(4-(2-((2-(3-fluoro-4-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro-
[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidaz-
olidine-2,4-dione;
1-(4-(2-((2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-4-oxo-1,3,8-triazaspir-
o[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimida-
zolidine-2,4-dione;
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione;
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2-
,4-dione);
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3-
,8-triazaspiro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.-
4]nonane-2,4-dione;
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-8-methyl-1,3,8-triazaspiro[-
4.5]decane-2,4-dione;
5-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2-oxa-5,7-diazaspiro[3.4]oc-
tane-6,8-dione; and
4-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-4,6-diazaspiro[2.4]heptane--
5,7-dione.
7. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein the compound is
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione
8. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein the compound is
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2-
,4-dione.
9. The compound or pharmacologically acceptable salt, or a hydrate
or a solvate thereof of claim 1, wherein the compound is
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2-
,4-dione.
10. A pharmaceutical composition, which comprises the compound or
pharmacologically acceptable salt, or a hydrate or a solvate
thereof of claim 1 as an active ingredient.
11. The pharmaceutical composition of claim 10, which is for use in
oral administration.
12. A pharmaceutical composition for activating intracellular cAMP
response, which comprises the compound or pharmacologically
acceptable salt, or a hydrate or a solvate thereof of claim 1 as an
active ingredient.
13. A stem cell-mobilizing agent, or an agent for preventing or
treating osteoporosis, fracture, adynamic bone disease,
achondroplasia, hypochondroplasia, osteomalacia, osteoarthritis,
arthritis, thrombocytopenia, hypoparathyroidism, hyperphosphatemia
or tumoral calcinosis, which comprises the compound or
pharmacologically acceptable salt, or a hydrate or a solvate
thereof of claim 1 as an active ingredient.
14. A method for prevention or treatment of osteoporosis, fracture,
adynamic bone disease, achondronplasia, hypochondroplasia,
osteomalacia, osteoarthritis, arthritis, thrombocytopenia,
hypoparathyroidism, hyperphosphatemia or tumoral calcinosis, or
stem cell mobilization, wherein the method comprises administering
a pharmaceutically effective amount of a composition comprising the
compound or pharmacologically acceptable salt, or a hydrate or a
solvate thereof of claim 1 to a patient in need of the prevention
or treatment of the disease or stem cell mobilization.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation of U.S. application Ser.
No. 15/214,729, filed Jul. 20, 2016, which is a Continuation of
U.S. application Ser. No. 14/426,408, which issued as U.S. Pat. No.
9,428,505, on Aug. 30, 2016, and which is the U.S. National Stage
application of PCT/JP2013/083022, filed Dec. 10, 2013, which claims
priority from Japanese Application No. 2012-269178, filed Dec. 10,
2012.
TECHNICAL FIELD
[0002] The present invention relates to pharmaceuticals comprising
as an active ingredient a hydantoin derivative that has high
metabolic stability and exhibits a potent PTH-like effect.
BACKGROUND ART
[0003] Parathyroid hormone (PTH) binds to the PTH1 receptor
(PTH1R), which is a G protein-coupled receptor (GPCR), to activate
the G protein, and then causes activation of at least one signaling
cascade such as the cyclic AMP (cAMP)/protein kinase A cascade. PTH
is known as a hormone that acts on target cells in the kidney and
bone to regulate calcium (Ca) and phosphorus (Pi) homeostasis
(Non-Patent Document 1). Serum Ca concentration level is maintained
by PTH mainly through direct or indirect actions on the
gastrointestinal tract, bone, and kidney. PTH promotes resorption
of Ca from the renal tubules and thereby suppresses excretion of Ca
in the body to the outside. It also increases the synthesis of an
enzyme that converts vitamin D to active vitamin D in the kidney,
and thereby contributes to the facilitation of active vitamin
D-mediated Ca absorption from the gastrointestinal tract.
Furthermore, PTH enhances the differentiation of osteoclasts
indirectly via osteoblasts and promote Ca release from the bone.
These actions of PTH are thought to occur mainly via the cyclic
adenosine 3',5'-monophosphate (cAMP) elevation and/or phospholipase
C (PLC) activation that occurs when PTH binds to the PTH1R.
[0004] In humans, PTH preparations [PTH (1-34) and PTH (1-84)] have
a powerful osteogenic effect, and induce significant increases in
bone mineral density (BMD) and bone strength. Currently, most of
the osteoporosis drugs available for humans are inhibitors of bone
resorption, and the only type of osteogenic drug that actively
increases BMD is PTH preparations. PTH preparation is regarded as
one of the most effective treatments for osteoporosis (Non-Patent
Document 2); however, since it is a peptide, it needs to be
administered by an invasive method. Therefore, there is an
expectation for production of a pharmaceutical agent that has
PTH-like effects and which can be administered non-invasively.
[0005] Hypoparathyroidism is a metabolic disease that exhibits
hypocalcemia and hyperphosphatemia caused by insufficiency of PTH
secreted from the parathyroid gland, and a variety of associated
symptoms. Active vitamin D preparations and Ca agents are being
used for the treatment of hypoparathyroidism; however, since the
PTH-mediated regulatory mechanism does not work, a sufficient
therapeutic effect is not obtained. Furthermore, since active
vitamin D formulations enhance urinary Ca excretion, long-term
therapy suggests an increased risk of nephropathy. In order to
solve these problems, there is an ongoing investigation of
replacement therapy that uses PTH preparations against this
disease; and an attempt was made to carry out several invasive
administrations per day or a continuous administration using a pump
to obtain sufficient efficacy (Non-Patent Document 3). Therefore,
for hypoparathyroidism treatment, generation of a pharmaceutical
agent that has PTH-like effects and which can also be administered
non-invasively is desirable.
[0006] Also, a pharmaceutical agent having PTH-like effects that
can also be administered non-invasively is desired for treatment of
diseases such as fracture, adynamic bone disease, achondroplasia,
hypochondroplasia, osteomalacia, osteoarthritis, arthritis,
thrombocytopenia, hyperphosphatemia, and tumoral calcinosis.
[0007] Under such circumstances, the present inventors submitted a
patent application in advance based on their discovery that the
compound represented by formula (A):
##STR00002##
[Patent Document 1 may be referred to for W, X, Y, m, n, R.sub.1,
R.sub.2, R.sub.33, and R.sub.34 in the formula] and
pharmacologically acceptable salts thereof are useful as compounds
having PTH-like effects, or more preferably, as a PTH1R agonist,
and are useful for prevention and/or treatment of osteoporosis,
fracture, osteomalacia, arthritis, thrombocytopenia,
hypoparathyroidism, hyperphosphatemia, or tumoral calcinosis, or
stem cell mobilization (Patent Document 1).
[0008] To produce pharmaceutical agents that have high clinical
value and can be administered invasively, it is necessary to
consider the in vivo kinetics such as absorption, distribution,
metabolism, and excretion of the drug in addition to its direct
actions on the target. To enable oral administration in particular,
it is desirable to have a pharmaceutical agent having PTH-like
effects which are high metabolic stability against human liver
microsomes and strong human PTH1R-mediated ability of producing
cAMP.
[0009] To provide a pharmaceutical agent that can be administered
orally to humans, generally a method of confirming the effects of
oral administration by in vivo testing that involves use of a model
animal. For example, a thyroparathyroidectomized (TPTX) rat is
known as an animal model for hypoparathyroidism. To find a
therapeutic agent that has strong PTH-like effects and high
metabolic stability, and works against hypoparathyroidism when
administered orally, it is effective to use a method of finding a
compound that acts on rat PTH1R and is stable to the rat's
metabolic enzymes, and then examining its actions when orally
administered to a TPTX rat model.
[0010] In current therapy for hypoparathyroidism, the therapeutic
target range for serum Ca concentration is set to a slightly lower
range than the lower limit of the normal range at 7.6 to 8.8 mg/dL
(Non-Patent Document 4). Since the normal range for rat serum Ca
concentration is the same level as for humans at 10 mg/dL or so, to
verify the therapeutic effect, it is important to attain a serum Ca
concentration in the rat model of the disease within the range from
the therapeutic target range in humans (7.6-8.8 mg/dL) to the lower
limit for hypercalcemia in humans (approximately 11.2 mg/dL).
PRIOR ART DOCUMENTS
Patent Documents
[0011] [Patent document 1] WO 2010/126030
Non-Patent Documents
[0011] [0012] [Non-patent document 1] Kronenberg, H. M., et al., In
Handbook of Experimental Pharmacology, Mundy, G. R., and Martin, T.
J., (eds), pp. 185-201, Springer-Verlag, Heidelberg (1993) [0013]
[Non-patent document 2] Tashjian and Gagel, J. Bone Miner. Res.
21:354-365 (2006) [0014] [Non-patent document 3] Rejnmark et al.,
Osteoporosis Int. Published Online: 27 Nov. 2012 [0015] [Non-patent
document 4] Winer K K et al., J. Clin. Endocrinol. Metab.
88(9):4214-4220 (2003)
SUMMARY OF THE INVENTION
Problems to be Solved by the Invention
[0016] An objective of the present invention is to discover
compounds with strong PTH-like effects and high metabolic
stability, and to provide pharmaceutical compositions comprising
such compounds to enable treatment of conditions that may be
treated by PTH-like actions, such as hypoparathyroidism.
Means for Solving the Problems
[0017] Under such circumstances, the present inventors continued to
carry out research, and discovered that the newly discovered
hydantoin derivatives of the present invention show strong
cAMP-producing ability in cells expressing human PTH1R, and have
high stability in human liver microsomes. The present inventors
also discovered that the compounds of the present invention show
strong cAMP-producing ability in cells expressing rat PTH1R, and
have high stability in rat hepatocytes. Additionally, in TPTX rat
models subjected to oral administration, it was newly discovered
that a dose of 30 mg/kg restored the serum Ca concentration to the
therapeutic target range of 7.6-8.8 mg/dL. Results obtained from
these model animals suggest that the compounds represented by
formula (1), which show a strong effect on human PTH1R and high
stability in human liver microsomes, are useful as therapeutic
agents for hypoparathyroidism.
[0018] The present invention relates to the following:
[1] A compound represented by general formula (1) below or a
pharmacologically acceptable salt thereof:
##STR00003##
(wherein, when R1 and R2 are not both hydrogen atoms, R1 and R2 are
independently: 1) hydrogen atom; 2) halogen atom; 3) an alkyl group
comprising one or two carbons that may be substituted with one to
five fluorine atoms; or 4) an alkoxy group comprising one or two
carbons that may be substituted with one to five fluorine atoms; or
R1 and R2 bond with each other to form a group represented by the
formula below:
##STR00004##
(wherein each * indicates the position of bonding with the phenyl
portion); and
[0019] R3 and R4 are independently a methyl group that may be
substituted with one to three fluorine atoms; or
[0020] R3 and R4, together with a bound carbon atom, form a three-
to six-membered carbocyclic ring (wherein, one of the carbon atoms
forming the ring may be replaced with an oxygen atom, a sulfur
atom, or a methyl-substituted or unsubstituted nitrogen atom).
[0021] In the present invention, a compound in which the
combination of R1 and R2 is a trifluoromethyl group and a hydrogen
atom, and where R3 and R4, together with a bound carbon atom, form
a cyclopentyl ring, can be excluded from the above-mentioned
compounds represented by formula (1).
[2] The compound or pharmacologically acceptable salt thereof of
[1], wherein R1 and R2 are selected from the combinations
below:
[0022] 1) R1 is a hydrogen atom or a halogen atom, and R2 is a
hydrogen atom, a trifluoromethyl group, or a trifluoromethoxy group
(provided that R1 and R2 are not both hydrogen atoms);
[0023] 2) R1 is a trifluoromethyl group or a trifluoromethoxy
group, and R2 is a hydrogen atom or a halogen atom;
[0024] 3) R1 and R2 bond with each other to form a group
represented by the formula below:
##STR00005##
(wherein, each * indicates the position of bonding with the phenyl
portion); and
[0025] R3 and R4 are methyl groups; or
[0026] R3 and R4, together with a bound carbon atom, form a ring
selected from below:
##STR00006##
(wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion). [3] The compound or
pharmacologically acceptable salt thereof of [1], wherein R1 and R2
are selected from the combinations below:
[0027] 1) R1 is a trifluoromethoxy group and R2 is a fluorine
atom;
[0028] 2) R1 is a bromine atom and R2 is a hydrogen atom;
[0029] 3) R1 is a trifluoromethoxy group and R2 is a fluorine
atom;
[0030] 4) R1 is a fluorine atom and R2 is a trifluoromethoxy
group;
[0031] 5) R1 is a trifluoromethyl group and R2 is a hydrogen
atom;
[0032] 6) R1 is a hydrogen atom and R2 is a trifluoromethoxy
group;
[0033] 7) R1 and R2 bond with each other to form a group
represented by the formula below:
##STR00007##
(wherein each * indicates the position of bonding with the phenyl
portion); and
[0034] R3 and R4 are methyl groups; or
[0035] R3 and R4, together with a bound carbon atom, form a ring
selected from below:
##STR00008##
(wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion). [4] The compound or
pharmacologically acceptable salt thereof of [1], wherein R3 and R4
are methyl groups. [5] The compound or pharmacologically acceptable
salt thereof of [1], wherein R3 and R4, together with a bound
carbon atom, form a ring selected from below:
##STR00009##
(wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion). [6] The compound or
pharmacologically acceptable salt thereof of [1], wherein the
compound is selected from the group consisting of: [0036]
1-(4-(2-((2-(4-fluoro-3-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro-
[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidaz-
olidine-2,4-dione; [0037]
1-(4-(2-((2-(3-bromophenyl)-4-oxo-1,3,8-triazaspiro[4.5]deca-1-en-8-yl)su-
lfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidazolidine-2,4-dione;
[0038]
1-(4-(2-((2-(4-fluoro-3-(trifluoromethyl)phenyl)-4-oxo-1,3,8-triaz-
aspiro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethyl-
imidazolidine-2,4-dione; [0039]
1-(4-(2-((2-(3-fluoro-4-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro-
[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidaz-
olidine-2,4-dione; [0040]
1-(4-(2-((2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-4-oxo-1,3,8-triazaspir-
o[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimida-
zolidine-2,4-dione; [0041]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione; [0042]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2-
,4-dione); [0043]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2-
,4-dione; [0044]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-8-methyl-1,3,8-triazaspiro[-
4.5]decane-2,4-dione; [0045]
5-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2-oxa-5,7-diazaspiro[3.4]oc-
tane-6,8-dione; and [0046]
4-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-4,6-diazaspiro[2.4]heptane--
5,7-dione. [7] The compound or pharmacologically acceptable salt
thereof of [1], wherein the compound is [0047]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione [8] The
compound or pharmacologically acceptable salt thereof of [1],
wherein the compound is
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2-
,4-dione. [9] The compound or pharmacologically acceptable salt
thereof of [1], wherein the compound is
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2-
,4-dione. [10] A pharmaceutical composition, which comprises the
compound or pharmacologically acceptable salt thereof of any one of
[1] to [9] as an active ingredient. [11] The pharmaceutical
composition of [10], which is for use in oral administration. [12]
A pharmaceutical composition for activating intracellular cAMP
response, which comprises the compound or pharmacologically
acceptable salt thereof of any one of [1] to [9] as an active
ingredient. [13] A stem cell-mobilizing agent, or an agent for
preventing or treating osteoporosis, fracture, adynamic bone
disease, achondroplasia, hypochondroplasia, osteomalacia,
osteoarthritis, arthritis, thrombocytopenia, hypoparathyroidism,
hyperphosphatemia or tumoral calcinosis, which comprises the
compound or pharmacologically acceptable salt thereof of any one of
[1] to [9] as an active ingredient. [14] A method for prevention or
treatment of osteoporosis, fracture, adynamic bone disease,
achondronplasia, hypochondroplasia, osteomalacia, osteoarthritis,
arthritis, thrombocytopenia, hypoparathyroidism, hyperphosphatemia
or tumoral calcinosis, or stem cell mobilization, wherein the
method comprises administering a pharmaceutically effective amount
of a composition comprising the compound or pharmacologically
acceptable salt thereof of any of [1] to [9] to a patient in need
of the prevention or treatment of the disease or stem cell
mobilization. [15] Use of the compound or pharmacologically
acceptable salt thereof of any one of [1] to [9] for the production
of a stem cell-mobilizing agent or an agent for preventing or
treating osteoporosis, fracture, adynamic bone disease,
achondroplasia, hypochondroplasia, osteomalacia, osteoarthritis,
arthritis, thrombocytopenia, hypoparathyroidism, hyperphosphatemia,
or tumoral calcinosis. [16] The compound or pharmacologically
acceptable salt thereof of any of [1] to [9] for treatment or
prevention of osteoporosis, fracture, adynamic bone disease,
achondronplasia, hypochondroplasia, osteomalacia, osteoarthritis,
arthritis, thrombocytopenia, hypoparathyroidism, hyperphosphatemia,
or tumoral calcinosis, or stem cell mobilization.
[0048] Furthermore the present invention provides methods for
treating pathological conditions that may be treated by PTH-like
actions, such as hypoparathyroidism, by administering a compound of
formula (1) or a salt thereof.
Effects of the Invention
[0049] The present invention provides hydantoin derivatives with
strong PTH-like effects and high metabolic stability. Use of the
hydantoin derivatives enables treatment of pathological conditions
caused by PTH-like actions, such as hypoparathyroidism.
BRIEF DESCRIPTION OF THE DRAWINGS
[0050] The FIGURE depicts a graph showing the average change of
level in serum Ca concentration for each compound up to 24 hours
after administration, when the compound is orally administered at a
dose of 30 mg/kg to a TPTX rat model.
MODE FOR CARRYING OUT THE INVENTION
[0051] The present invention relates to hydantoin derivatives and
use thereof. The present inventors have synthesized a compound
represented by the above formula (1) or a pharmacologically
acceptable salt thereof for the first time and have found that the
compound or a salt thereof is a compound having a strong
parathyroid hormone (PTH)-like effect and high metabolic
stability.
[0052] The "alkyl" herein refers to a monovalent group derived by
removing any one hydrogen atom from an aliphatic hydrocarbon, and
covers a subset of hydrocarbyl or hydrocarbon group structures not
containing a heteroatom or an unsaturated carbon-carbon bond and
containing hydrogen and carbon atoms in the backbone. Examples of
the alkyl group include those of linear or branched structures. The
alkyl group is preferably an alkyl group comprising one or two
carbon atoms. The alkyl group is specifically, for example, a
methyl group or an ethyl group, and is preferably a methyl
group.
[0053] The term "alkoxy" as used herein refers to an oxy group to
which the above-defined "alkyl" is bound, and preferably refers to
an alkoxy group comprising one or two carbon atoms. Specific
examples include methoxy and ethoxy groups, and a preferred example
is methoxy group.
[0054] The "B optionally substituted with A" herein denotes that
any hydrogen atom(s) in B may be substituted with any number of
As.
[0055] In the present invention, the number of substituents is not
limited unless otherwise indicated. For example, the number of
substituents may be 1 to 5, 1 to 4, 1 to 3, 1 to 2, or 1.
[0056] The "halogen atom" herein refers to a fluorine atom, a
chlorine atom, a bromine atom or an iodine atom.
[0057] Herein, the symbol "*" in the chemical formula refers to the
position of bonding.
[0058] Compounds of the present invention represented by formula
(1) has strong PTH-like effects and high metabolic stability.
[0059] The "PTH-like effect" herein refers to activity of
generating intracellular cAMP (cAMP: cyclic adenosine
monophosphate) by action on the PTH receptor or action on the
signal transduction pathway through the PTH receptor.
[0060] In the present invention, whether there is a "strong
PTH-like effect" or whether "a PTH-like effect is strong" can be
confirmed by measuring the cAMP signaling activity by analyzing
cAMP signaling, for example, according to the method described in
J. Bone. Miner. Res. 14:11-20, 1999. Specifically, for example,
according to the method described in Test Example 1, the amount of
cAMP produced in cells forced to express human PTH1R is determined
using a commercially available cAMP EIA kit (for example, Biotrack
cAMP EIA system, GE health care) to measure the concentration of
each compound at 20% cAMP signaling activity (EC20) or their
concentration at 50% cAMP signaling activity (EC50), with the cAMP
signaling activity obtained upon administration of 100 nM of human
PTH (1-34) being defined as 100%. In the present invention, for a
"strong PTH-like effect" or "a PTH-like effect is strong", for
example, the EC20 value (.mu.M) measured by the above-mentioned
method is preferably 5.0 or less, more preferably 3.0 or less, and
even more preferably 2.0 or less. For EC50, the value (.mu.M)
measured by the above-mentioned method is, for example, preferably
25.0 or less, more preferably 15.0 or less, and even more
preferably 10.0 or less.
[0061] Whether there is "high metabolic stability" or whether the
"metabolic stability is high" can be confirmed using a general
measurement method. For example, liver cells, small intestinal
cells, liver microsomes, small intestinal microsomes, liver S9, and
such may be used for the confirmation. Specifically, for example,
the stability of a compound in liver microsomes can be confirmed by
taking measurements according to description in T. Kronbach et al.
(Oxidation of midazolam and triazolam by human liver cytochrome
P450IIIA4. Mol. Pharmacol, 1989, 36(1), 89-96). More specifically,
the stability can be confirmed by following the method described in
Test Example 3. In the present invention, "high metabolic
stability" or "metabolic stability is high" are when the clearance
(L/min/mg) value in the metabolic stability test using human liver
microsomes described in the above-mentioned Test Example is
preferably 60 or less, more preferably 40 or less, and even more
preferably 35 or less. Specifically, high metabolic stability can
be obtained in the aforementioned formula (1), except where the
combination of R1 and R2 is a trifluoromethyl group and a hydrogen
atom, and R3 and R4, together with a bound carbon atom, form a
cyclopentyl ring.
[0062] The compounds according to the present invention, whether
free forms or pharmacologically acceptable salts, are included in
the present invention. Examples of such "salts" include inorganic
acid salts, organic acid salts, inorganic base salts, organic base
salts and acidic or basic amino acid salts.
[0063] Preferred examples of the inorganic acid salts include
hydrochlorides, hydrobromides, sulfates, nitrates and phosphates.
Preferred examples of the organic acid salts include acetates,
succinates, fumarates, maleates, tartrates, citrates, lactates,
stearates, benzoates, methanesulfonates, benzenesulfonates, and
p-toluenesulfonates.
[0064] Preferred examples of the inorganic base salts include
alkali metal salts such as sodium salts and potassium salts,
alkaline earth metal salts such as calcium salts and magnesium
salts, aluminum salts and ammonium salts. Preferred examples of the
organic base salts include diethylamine salts, diethanolamine
salts, meglumine salts and N,N-dibenzylethylenediamine salts.
[0065] Preferred examples of the acidic amino acid salts include
aspartates and glutamates. Preferred examples of the basic amino
acid salts include arginine salts, lysine salts and ornithine
salts.
[0066] The compounds of the present invention may absorb moisture,
have adsorbed water or form hydrates when left in the air. Such
hydrates are also included in the salts of the present
invention.
[0067] Further, the compounds of the present invention may absorb
certain other solvents to form solvates. Such salts are also
encompassed in the present invention as salts of the compounds of
the formula (1).
[0068] Herein, a structural formula of a compound may represent a
certain isomer for the sake of convenience. However, the compounds
of the present invention include all isomers such as geometric
isomers, optical isomers based on asymmetric carbons, stereoisomers
and tautomers as well as mixtures of these isomers which occur due
to the structures of the compounds, without being limited to the
formulas described for the sake of convenience, and may be either
one of isomers or a mixture thereof. Thus, the compounds of the
present invention may have an asymmetric carbon atom in the
molecule and may be present as optically active forms and
racemates, but the present invention is not limited to either of
them and includes both of them.
[0069] The present invention includes all isotopes of the compounds
represented by the formula (1). In the isotopes of the compounds of
the present invention, at least one atom is replaced by an atom
having the same atomic number (proton number) but having a
different mass number (sum of the number of protons and the number
of neutrons). Examples of the isotopes contained in the compounds
of the present invention include a hydrogen atom, a carbon atom, a
nitrogen atom, an oxygen atom, a phosphorus atom, a sulfur atom, a
fluorine atom and a chlorine atom, including .sup.2H, .sup.3H,
.sup.13C, .sup.14C, .sup.15N, .sup.17O, .sup.18O, .sup.31P,
.sup.32P, .sup.35S, .sup.18F and .sup.36Cl, respectively. In
particular, radioisotopes that decay by emitting radioactivity such
as .sup.3H and .sup.14C are useful in body tissue distribution
tests for pharmaceuticals or compounds. Stable isotopes do not
decay, are almost equal in abundance and do not emit radioactivity,
and thus they can be used safely. The isotopes of the compounds of
the present invention can be converted according to conventional
methods by substituting a reagent containing a corresponding
isotope for a reagent used for synthesis.
[0070] The compounds according to the present invention may exhibit
crystalline polymorphism, but are not particularly limited to any
one of these, but may be in any one of these crystal forms or exist
as a mixture of two or more crystal forms.
[0071] The compounds according to the present invention include
prodrugs thereof. The prodrugs are derivatives of the compounds of
the present invention which have chemically or metabolically
decomposable groups and are converted back to the original
compounds after administration in vivo to exhibit their original
efficacy, including complexes not formed with covalent bonds, and
salts.
[0072] The compounds represented by the above formula (1) according
to the present invention are preferably as follows.
[0073] In the formula, R1 and R2 are selected from the combinations
below:
[0074] 1) R1 is a hydrogen atom or a halogen atom, and R2 is a
hydrogen atom, a trifluoromethyl group, or a trifluoromethoxy group
(provided that R1 and R2 are not both hydrogen atoms);
[0075] 2) R1 is a trifluoromethyl group or a trifluoromethoxy
group, and R2 is a hydrogen atom or a halogen atom;
[0076] 3) R1 and R2 bond with each other to form a group
represented by the formula below:
##STR00010##
(wherein, * each indicates the position of bonding with the phenyl
portion); and
[0077] R3 and R4 are methyl groups; or
[0078] R3 and R4, together with a bound carbon atom, form a ring
selected from below:
##STR00011##
(wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion).
[0079] The compounds represented by the above formula (1) according
to the present invention are more preferably as follows.
[0080] In the formula, R1 and R2 are selected from the combinations
below:
[0081] 1) R1 is a trifluoromethoxy group and R2 is a fluorine
atom;
[0082] 2) R1 is a bromine atom and R2 is a hydrogen atom;
[0083] 3) R1 is a trifluoromethoxy group and R2 is a fluorine
atom;
[0084] 4) R1 is a fluorine atom and R2 is a trifluoromethoxy
group;
[0085] 5) R1 is a trifluoromethyl group and R2 is a hydrogen
atom;
[0086] 6) R1 is a hydrogen atom and R2 is a trifluoromethoxy
group;
[0087] 7) R1 and R2 bond with each other to form a group
represented by the formula below:
##STR00012##
(wherein * each indicates the position of bonding with the phenyl
portion); and
[0088] R3 and R4 are methyl groups; or
[0089] R3 and R4, together with a bound carbon atom, form a ring
selected from below:
##STR00013##
(wherein * indicates the position of bonding with the
imidazolidine-2,4-dione portion).
[0090] The compounds represented by the above formula (1) according
to the present invention are further preferably a compound selected
from the group consisting of the following, or a pharmacologically
acceptable salt thereof.
Compound 1:
[0091]
1-(4-(2-((2-(4-fluoro-3-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-tri-
azaspiro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimeth-
ylimidazolidine-2,4-dione;
Compound 2:
[0091] [0092]
1-(4-(2-((2-(3-bromophenyl)-4-oxo-1,3,8-triazaspiro[4.5]deca-1-en-8-yl)su-
lfonyl)ethyl)-3,5-dim
ethylphenyl)-5,5-dimethylimidazolidine-2,4-dione;
Compound 3:
[0092] [0093]
1-(4-(2-((2-(4-fluoro-3-(trifluoromethyl)phenyl)-4-oxo-1,3,8-triazaspiro[-
4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidazo-
lidine-2,4-dione;
Compound 4:
[0093] [0094]
1-(4-(2-((2-(3-fluoro-4-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro-
[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidaz-
olidine-2,4-dione;
Compound 5:
[0094] [0095]
1-(4-(2-((2-(2,2-difluorobenzo[d][1,3]dioxol-5-yl)-4-oxo-1,3,8-triazaspir-
o[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimida-
zolidine-2,4-dione;
Compound 6:
[0095] [0096]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione;
Compound 7:
[0096] [0097]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2-
,4-dione);
Compound 8:
[0097] [0098]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2-
,4-dione;
Compound 9:
[0098] [0099]
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-8-methyl-1,3,8-triazaspiro[-
4.5]decane-2,4-dione;
Compound 10:
[0099] [0100]
5-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2-oxa-5,7-diazaspiro[3.4]oc-
tane-6,8-dione; and
Compound 11:
[0100] [0101]
4-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-4,6-diazaspiro[2.4]heptane--
5,7-dione.
[0102] Of Compounds 1 to 11 above, Compounds 6, 7 and 8 are more
preferred.
[0103] Such compounds represented by the above formula (1) or
pharmacologically acceptable salts thereof according to the present
invention are useful as compounds having a PTH-like effect,
preferably PTH1R agonists, and are useful for the prevention and/or
treatment of osteoporosis, fracture, adynamic bone disease,
achondronplasia, hypochondroplasia, osteomalacia, osteoarthritis,
arthritis, thrombocytopenia, hypoparathyroidism, hyperphosphatemia,
tumoral calcinosis or the like, or stem cell mobilization.
[0104] The compounds or salts thereof according to the present
invention can be formulated by conventional methods into tablets,
powders, fine granules, granules, coated tablets, capsules, syrups,
troches, inhalations, suppositories, injections, ointments,
ophthalmic ointments, ophthalmic preparations, nasal preparations,
ear preparations, cataplasms, lotions and the like. Commonly used
excipients, binders, lubricants, colorants, correctives, and as
necessary, stabilizers, emulsifiers, absorption promoters,
surfactants, pH adjusters, preservatives, antioxidants and the like
can be used for formulation, and they are blended with ingredients
commonly used as raw materials of pharmaceutical preparations and
formulated by conventional methods.
[0105] For example, oral preparations are manufactured by adding,
to the compound or a pharmacologically acceptable salt thereof
according to the present invention, an excipient, and as necessary,
a binder, a disintegrant, a lubricant, a colorant, a corrective and
the like and then formulating them into powder, fine granules,
granules, tablets, coated tablets, capsules and the like by a
conventional method.
[0106] Examples of these ingredients include animal and vegetable
oils such as soybean oil, beef tallow and synthetic glyceride;
hydrocarbons such as liquid paraffin, squalane and solid paraffin;
ester oils such as octyldodecyl myristate and isopropyl myristate;
higher alcohols such as cetostearyl alcohol and behenyl alcohol;
silicone resin; silicone oil; surfactants such as polyoxyethylene
fatty acid ester, sorbitan fatty acid ester, glycerol fatty acid
ester, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene
hydrogenated castor oil and a polyoxyethylene-polyoxypropylene
block copolymer; water-soluble polymers such as
hydroxyethylcellulose, polyacrylic acid, a carboxyvinyl polymer,
polyethylene glycol, polyvinylpyrrolidone and methylcellulose;
lower alcohols such as ethanol and isopropanol; polyhydric alcohols
such as glycerol, propylene glycol, dipropylene glycol and
sorbitol; sugars such as glucose and sucrose; inorganic powders
such as silicic anhydride, magnesium aluminum silicate and aluminum
silicate; and purified water.
[0107] Examples of the excipients include lactose, corn starch,
white soft sugar, glucose, mannitol, sorbitol, microcrystalline
cellulose and silicon dioxide.
[0108] Examples of the binders include polyvinyl alcohol, polyvinyl
ether, methylcellulose, ethylcellulose, acacia, tragacanth,
gelatin, shellac, hydroxypropylmethylcellulose,
hydroxypropylcellulose, polyvinylpyrrolidone, a polypropylene
glycol-polyoxyethylene block polymer and meglumine.
[0109] Examples of the disintegrants include starch, agar, gelatin
powder, microcrystalline cellulose, calcium carbonate, sodium
bicarbonate, calcium citrate, dextrin, pectin and
carboxymethylcellulose calcium.
[0110] Examples of the lubricants include magnesium stearate, talc,
polyethylene glycol, silica and hydrogenated vegetable oil.
[0111] Colorants used are those approved as additives to
pharmaceuticals. Correctives used are cocoa powder, peppermint
camphor, empasm, mentha oil, borneol, powdered cinnamon bark and
the like.
[0112] Obviously, these tablets and granules may be sugar-coated or
otherwise coated appropriately as necessary. Liquid preparations
such as syrups and injectable preparations are manufactured by
adding a pH adjuster, a solubilizer, a tonicity adjusting agent and
the like, and as necessary, a solubilizing agent, a stabilizer and
the like to the compound or a pharmacologically acceptable salt
thereof according to the present invention and formulating them by
a conventional method.
[0113] The method of manufacturing external preparations is not
limited and they can be manufactured by conventional methods.
Specifically, various raw materials commonly used for
pharmaceuticals, quasi drugs, cosmetics and the like can be used as
base materials for formulation. Specific examples of the base
materials used include raw materials such as animal and vegetable
oils, mineral oils, ester oils, waxes, higher alcohols, fatty
acids, silicone oil, surfactants, phospholipids, alcohols,
polyhydric alcohols, water-soluble polymers, clay minerals and
purified water. Further, pH adjusters, antioxidants, chelators,
preservatives and fungicides, colorants, flavors and the like may
be added as necessary. The base materials for external preparations
according to the present invention are not limited to these
materials.
[0114] Ingredients such as ingredients having a
differentiation-inducing effect, blood flow promoters,
bactericides, anti-inflammatory agents, cell activators, vitamins,
amino acids, humectants and keratolytic agents may also be blended
as necessary. The aforementioned base materials are added in an
amount corresponding to the concentration usually chosen for the
manufacture of external preparations.
[0115] The mode of administration of the compounds or salts
thereof, or hydrates of the compounds or salts according to the
present invention is not particularly limited, and they may be
orally or parenterally administered by methods commonly used. For
example, they can be formulated into preparations such as tablets,
powders, granules, capsules, syrups, troches, inhalations,
suppositories, injections, ointments, ophthalmic ointments,
ophthalmic preparations, nasal preparations, ear preparations,
cataplasms and lotions and administered.
[0116] The compounds of the present invention are particularly
suitable for formulation into oral agents since they show an
excellent cAMP signaling activity and have metabolic stability.
[0117] The dosage of the medicine according to the present
invention can be appropriately selected depending on the severity
of the symptom, the age, the sex, the body weight, the mode of
administration, the type of the salt, the specific type of the
disease, and the like.
[0118] Although the dosage significantly varies according to the
type of the disease and the severity of the symptom of the patient,
the age of the patient, the sex difference and the difference in
sensitivity to drugs between the patients, and the like, the dosage
is usually about 0.03 to 1000 mg, preferably 0.1 to 500 mg and more
preferably 0.1 to 100 mg per day for adults and is administered
divided into one to several doses a day.
[0119] In the manufacture of the compounds of the present invention
represented by the above formula (1), raw material compounds and
various reagents may form salts, hydrates or solvates, all vary
according to the starting material, the solvent used, and the like,
and are not particularly limited insofar as they do not inhibit the
reaction.
[0120] The solvent used also varies according to the starting
material, the reagent and the like, and is not particularly limited
insofar as it does not inhibit the reaction and dissolves the
starting material to a certain extent, obviously.
[0121] Various isomers (e.g., geometric isomers, optical isomers
based on asymmetric carbons, rotamers, stereoisomers and tautomers)
can be purified and isolated using common separation means, e.g.,
recrystallization, diastereomeric salt methods, enzymatic
resolution methods and various chromatography methods (e.g.,
thin-layer chromatography, column chromatography, high performance
liquid chromatography and gas chromatography).
[0122] The compounds according to the present invention obtained as
free forms can be converted to salts that may be formed by the
compounds or to hydrates of the compounds according to conventional
methods. The compounds according to the present invention obtained
as salts or hydrates of the compounds can also be converted to free
forms of the compounds according to conventional methods.
[0123] The compounds according to the present invention can be
isolated and purified by applying common chemical operations such
as extraction, concentration, evaporation, crystallization,
filtration, recrystallization and various chromatography
methods.
[0124] All prior art documents cited herein are hereby incorporated
by reference.
General Synthesis Methods
[0125] The compounds of the present invention can be synthesized by
various methods, some of which will be described with reference to
the following schemes. The schemes are illustrative and the present
invention is not limited only by the chemical reactions and
conditions explicitly indicated. Although some substituents are
excluded in the following schemes for the sake of clarity, such
exclusion is not intended to limit the disclosure of the schemes.
Representative compounds of the present invention can be
synthesized using appropriate intermediates, known compounds, and
reagents. R.sub.1, R.sub.2, R.sub.3 and R.sub.4 in the formulas in
the following general synthesis methods are as defined for R.sub.1,
R.sub.2, R.sub.3 and R.sub.4 in the compounds represented by the
above general formula (1) (compounds represented by formula 1 in
the following general synthesis methods).
[0126] The compounds of the present invention (Formula 1) can be
synthesized by the manufacturing methods (Methods A and B) shown
below.
##STR00014##
[0127] Scheme 1 shows a method for obtaining a hydantoin derivative
(Formula 1) by (1)amidation of thecarboxylic acid derivative (1)
and the amino-amide derivative (2) to obtain the amide-amide
derivative (3), and then constructing the spiroimidazolone ring by
intramolecular cyclization.
[0128] Step 1 is a method of the amidation of a carboxylic acid
derivative (1) and an hexafluorophosphate (HATU) and
4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride
n-hydrate (DMT-MM). Examples of the base include triethylamine or
N,N-diisopropylethylamine. If necessary, 4-(dimethylamino)pyridine
(DMAP) may be used as a catalyst. Examples of the appropriate
solvent include dichloromethane or N,N-dimethylformamide. Examples
of the appropriate reaction solvent when DMT-MM is used include
methanol, ethanol and acetonitrile. The reaction temperature is
0.degree. C. to room temperature, for example, and the reaction
time is 0.5 to 24 hours. The resulting amino-amide derivative (3)
is isolated by a common technique and, if necessary, may be
purified by crystallization or chromatography.
[0129] Step 2 is a method for the cyclization of the amide-amide
derivative (3) in the presence of a suitable base such as an
aqueous sodium hydroxide solution or potassium t-butoxide in a
suitable solvent such as ethanol, tert-butanol, or
dimethylsulfoxide. The reaction temperature is carried out, for
example, under room temperature to refluxing conditions for one to
24 hours. The obtained hydantoin derivative (Formula 1) is isolated
by common techniques, and when necessary, it can be purified by
crystallization or chromatography.
[0130] The amino-amide derivative (2) indicated in Scheme 1 can be
synthesized from the piperidine derivative (4). The synthetic
method for the amino-amide derivative (2) is shown in Scheme 2.
##STR00015##
[0131] Step 3 is a Strecker synthesis of converting a piperidinone
derivative (4) to an amino-nitrile derivative (5). Specifically,
this is a method of reacting a piperidinone (4) with sodium cyanide
or potassium cyanide and ammonium chloride or ammonium acetate in
an appropriate solvent such as methanol, ethanol or tetrahydrofuran
in the presence/absence of water. The reaction temperature is room
temperature to 80.degree. C., for example, and the reaction time is
2 to 72 hours. The resulting amino-nitrile derivative (5) is
isolated by a common technique and, if necessary, may be purified
by crystallization or chromatography.
[0132] Step 4 is a method of converting the nitrile group to an
amido group under basic hydrolysis conditions in the presence of
hydrogen peroxide. This reaction can be performed with reference to
Chemistry-A European Journal (2002), 8(2), 439-450, for
example.
[0133] Step 5 is a method of the hydrogenation of an olefin
Compound (6) in an inert solvent such as methanol, ethanol,
trifluoroethanol, dimethylformamide or dimethylacetamide in the
presence of a catalyst such as palladium carbon or palladium
hydroxide carbon, respectively, under an H.sub.2 atmosphere. The
reaction temperature is room temperature to 80.degree. C., and the
reaction may be performed under pressure. The resulting amino-amide
derivative (2) is isolated by a common technique and, if necessary,
may be purified by crystallization or chromatography.
[0134] The piperidinone derivative (4) shown in Scheme 2 can be
synthesized from a known ketal vinylsulfonyl derivative (7) and a
hydantoin-arylbromide derivative (8). The synthetic method for the
piperidine derivative (4) is shown in Scheme 3.
##STR00016##
[0135] Step 6 is a method for the synthesis of a
ketal-arylvinylsulfonyl derivative (9) by coupling the ketal
vinylsulfonyl derivative (7) and the hydantoin-arylbromide
derivative (8) under N.sub.2 atmosphere in the presence of a
palladium catalyst such as tris(dibenzilidineacetone)palladium(0)
or bis(dibenzylidineacetone)palladium, and by adding a phosphine
ligand such as tri-tert-butylphosphine tetrafluoroboric acid and a
suitable base such as methyldicyclohexylamine, in a suitable
solvent such as N-methyl-2-piperidone (NMP). The reaction
temperature is between 90.degree. C. and refluxing temperature. The
obtained ketal-arylvinylsulfonyl derivative (9) is isolated by
common techniques, and when necessary, it can be purified by
crystallization or chromatography.
[0136] Step 7 is a method for the conversion of ketal of the
ketal-arylvinylsulfonyl derivative (9) to ketone in a suitable
solvent such as aqueous tetrahydrofuran in the presence of an acid
such as hydrochloric acid. The reaction temperature is, for
example, the boiling point of the solvent, and the reaction time is
approximately 1 to 24 hours. The obtained piperidine derivative (4)
is isolated by common techniques, and when necessary, it can be
purified by crystallization or chromatography.
[0137] The hydantoin-arylbromide derivative (8) shown in Scheme 3
can be synthesized from 4-bromo-3,5-dimethylaniline (10) and the
bromoacetic acid derivative (11), or from
2-bromo-5-iodo-1,3-dimethylbenzene (13) and the amino acid
derivative (14). A synthetic method for the hydantoin-aryl bromide
derivative (8) is shown in Scheme 4.
##STR00017##
[0138] Step 8 is a method for the alkylation of
4-bromo-3,5-dimethylaniline (10) with the bromoacetic acid
derivative (11) in the presence of a suitable base such as
diisopropylethylamine and in a suitable solvent such as
N-methyl-2-piperidone (NMP). The reaction temperature is, for
example, room temperature to 100.degree. C., and the reaction time
is 1 to 24 hours. The obtained arylbromide-amino acid derivative
(12) is isolated by common techniques, and when necessary, it can
be purified by crystallization or chromatography.
[0139] Step 9 is a method for the synthesis of the
arylbromide-amino acid derivative (12), by coupling of
2-bromo-5-iodo-1,3-dimethylbenzene (13) and the amino acid
derivative (14) in the presence of a metal catalyst such as copper
iodide (I). The reaction can be carried out in the presence of a
suitable base such as diazabicycloundecene (DBU) and in a suitable
solvent such as N,N-dimethylacetamide (DMA), at a reaction
temperature of about 80.degree. C. to 120.degree. C. The obtained
arylbromide-amino acid derivative (12) is isolated by common
techniques, and when necessary, it can be purified by
crystallization or chromatography.
[0140] Step 10 is a method for the synthesis of the
hydantoin-arylbromide derivative (8) by reacting the
arylbromide-amino acid derivative (12) with sodium cyanate under an
acidic condition. The solvent is, for example, a mixed solvent such
as acetic acid-dichloromethane; the reaction temperature is room
temperature to 60.degree. C.; and the reaction time is 1 to 24
hours. The obtained hydantoin-arylbromide derivative (8) may be
isolated by common techniques, and when necessary, it can be
purified by crystallization or chromatography.
[0141] The hydantoin-arylbromide derivative (8) shown in Scheme 3
can also be synthesized from 4-bromo-3,5-dimethylaniline (10) and a
ketone derivative (15). A synthetic method for the
hydantoin-arylbromide derivative (8) is shown in Scheme 5.
##STR00018##
[0142] Step 11 is Strecker synthesis which directs the ketone
derivative (15) to become an arylamino-nitrile derivative (16).
More specifically, it is a method that reacts the ketone derivative
(15) with 4-bromo-3,5-dimethylaniline (10) and trimethylsilyl
cyanide in a suitable solvent such as acetic acid. The reaction
temperature may be room temperature, and the reaction time is one
to three hours or so. The obtained arylamino-nitrile derivative
(16) is isolated by common techniques, and when necessary, it can
be purified by crystallization or chromatography.
[0143] Step 12 is a method for reacting the aryl amino-nitrile
derivative (16) with 2,2,2-trichloroacetylisocyanate in a suitable
solvent such as dichloromethane, and then synthesizing an
iminohydantoin derivative (17) by adding reagents such as methanol,
water, and triethylamine and allowing them to react under heating
conditions. The obtained iminohydantoin derivative (17) is isolated
by common techniques, and when necessary, it can be purified by
crystallization or chromatography.
[0144] Step 13 is a method for the conversion of the iminohydantoin
derivative (17) into the hydantoin-arylbromide derivative (8) under
an acidic condition. For example, the synthesis can be carried out
in an acetic acid-water solvent with heating at approximately
65.degree. C. for one to six hours or so. The obtained
hydantoin-arylbromide derivative (8) is isolated by common
techniques, and when necessary, it can be purified by
crystallization or chromatography.
[0145] Scheme 6 is a method for a Heck reaction of a
vinylsulfonamide derivative (18) and the hydantoin-arylbromide
derivative (8) in the presence of a metal catalyst, and then the
hydrogenation of olefin compound (19) to give the hydantoin
derivative (Formula 1).
##STR00019##
[0146] The hydantoin derivative (Formula 1) can be synthesized by
performing the reaction of Step 14 according to the method of Step
6 and the reaction of Step 15 according to the method of Step 5.
The obtained hydantoin derivative (Formula 1) is isolated by common
techniques, and when necessary, it can be purified by
crystallization or chromatography.
[0147] The vinylsulfonamide derivative (18) used in Step 14 can be
synthesized by referring to Schemes 2, 3, and 12 of
WO2010/126030(A1).
[0148] All prior art documents cited in this specification are
incorporated herein by reference.
EXAMPLES
[0149] The content of the present invention will be described in
more detail by the following examples and test example; however,
the present invention is not limited to the content of the examples
and test example. All starting materials and reagents were obtained
from commercial suppliers or synthesized using known methods.
.sup.1H-NMR spectra were measured using Mercury300 (manufactured by
Varian), ECP-400 (manufactured by JEOL) or 400-MR (manufactured by
Varian) with or without Me.sub.4Si as the internal standard
(s=singlet, d=doublet, t=triplet, brs=broad singlet, m=multiplet).
Mass spectrometry measurement was performed using a mass
spectrometer, ZQ2000 (manufactured by Waters), SQD (manufactured by
Waters) or 2020 (manufactured by Shimazu).
Example 1
1-(4-(2-((2-(4-fluoro-3-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro[-
4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidazo-
lidine-2,4-dione (Compound 1)
##STR00020##
[0151] To a solution of 4-bromo-3,5-dimethylaniline (3.47 g, 17.4
mmol) and diisopropylethylamine (5.3 mL, 30.4 mmol) in DMI (13 mL),
2-bromoisobutyric acid (3.86 g, 23.1 mmol) was added at room
temperature. The mixture was stirred at 100.degree. C. for one
hour. And then 2-bromoisobutyrate (496 mg, 2.97 mmol) and
diisopropylethylamine (0.8 mL, 4.59 mmol) was added and the mixture
was stirred at 100.degree. C. for one hour.
[0152] Methanol (52 mL) and a 5 N aqueous sodium hydroxide solution
(52 mL, 260 mmol) were added to the reaction mixture at room
temperature, and then this mixture was stirred at 75.degree. C. for
1.5 hours. The reaction mixture was cooled, followed by addition of
water and adjustment of the pH to 5 using a 1 N aqueous potassium
hydrogen sulfate solution, and then extracted using ethyl acetate.
The organic layer was washed with water, then dried over anhydrous
magnesium sulfate, and concentrated to yield
2-((4-bromo-3,5-dimethylphenyl)amino)-2-methyl propanoic acid as a
crude product (5.79 g).
[0153] MS(ESI) m/z=286, 288 (M+H)+
##STR00021##
[0154] To a mixture of
2-((4-bromo-3,5-dimethylphenyl)amino)-2-methyl propanoic acid (5.79
g of the compound obtained from Reaction 1-1) in dichloromethane
(62 mL) and acetic acid (62 mL), sodium cyanate (5.03 g, 59.8 mmol)
was added at room temperature. The mixture was stirred at room
temperature for three hours. A saturated solution of sodium
hydrogen carbonate (400 mL) was added to adjust the pH to 7-8 using
a 5 N aqueous sodium hydroxide, and this mixture was extracted with
ethyl acetate. The organic layer was dried over anhydrous magnesium
sulfate, and then concentrated under reduced pressure. The obtained
solid was washed sequentially with ethyl acetate-hexane and then
with dichloromethane-hexane to obtain
1-(4-bromo-3,5-dimethylphenyl)-5,5-dimethylimidazolidine-2,4-dione
(3.80 g, 66%).
[0155] MS(ESI) m/z=311, 313 (M+H)+
##STR00022##
[0156] A mixture of
8-(vinylsulfonyl)-1,4-dioxa-8-azaspiro[4.5]decane (431 mg, 1.85
mmol),
1-(4-bromo-3,5-dimethylphenyl)-5,5-dimethylimidazolidine-2,4-dione
(575 mg, 1.85 mmol), tris(dibenzylidineacetone)palladium(0) (508
mg, 0.55 mmol), tri-tert-butylphosphine tetrafluoroboric acid (165
mg, 0.55 mmol), and methyldicyclohexylamine (2.1 mL, 9.25 mmol) in
N-methyl-2-pyrrolidone (18.5 mL) was stirred under nitrogen
atmosphere at 110.degree. C. for two hours. The reaction mixture
was cooled, quenched with water, and then extracted with ethyl
acetate. The organic layer was washed with water and brine, dried
over anhydrous magnesium sulfate, and then concentrated under
reduced pressure. The residue was purified by amino-silica gel
column chromatography (dichloromethane-methanol) to afford
(E)-1-(4-(2-(1,4-dioxa-8-azaspiro[4.5]decan-8-ylsulfonyl)vinyl)-3,5-dimet-
hylphenyl)-5,5-dimeth ylimidazolidine-2,4-dione (584 mg, 68%).
[0157] MS(ESI) m/z=464 (M+H)+
##STR00023##
[0158] To a solution of
(E)-1-(4-(2-(1,4-dioxa-8-azaspiro[4.5]decan-8-ylsulfonyl)vinyl)-3,5-dimet-
hylphenyl)-5,5-dimeth ylimidazolidine-2,4-dione (1.2 g, 2.58 mmol)
in tetrahydrofuran (26 mL), a 2 N aqueous hydrochloric acid
solution (26 mL, 52 mmol) was added dropwise over ten minutes. The
mixture was stirred at 60.degree. C. for two hours. The reaction
mixture was cooled, followed by adjustment of its pH to 7 using a 2
N aqueous sodium hydroxide solution, and this mixture was extracted
with ethyl acetate. The organic layer was washed with brine, dried
over anhydrous magnesium sulfate, and then concentrated under
reduced pressure. The residue was purified by silica gel column
chromatography (dichloromethane-ethyl acetate) to afford
(E)-1-(3,5-dimethyl-4-(2-((4-oxopipedridin-1-yl)sulfonyl)vinyl)phenyl)-5,-
5-dimethylimidazolidine-2,4-dione (998 mg, 92%).
[0159] MS(ESI) m/z=420 (M+H)+
##STR00024##
[0160] To a solution of
(E)-1-(3,5-dimethyl-4-(2-((4-oxopipedridin-1-yl)sulfonyl)vinyl)phenyl)-5,-
5-dimethylimidazolidine-2,4-dione (994 mg, 2.37 mmol) in methanol
(24 mL), potassium cyanide (188 mg, 2.84 mmol) and ammonium acetate
(237 mg, 3.08 mmol) were added at room temperature. The mixture was
stirred at 60-70.degree. C. for three hours. The reaction mixture
was cooled, concentrated under reduced pressure, and then diluted
with ethyl acetate. The organic layer was washed with water and
brine, dried over anhydrous magnesium sulfate, and then
concentrated under reduced pressure. The residue was purified by
silica gel column chromatography (dichloromethane-ethyl acetate) to
afford
(E)-4-amino-1-((4-(5,5-dimethyl-2,4-dioxoimidazolidin-1-yl)-2,6-dimethyls-
tyryl)sulfonyl)piperidine-4-carbonitrile (681 mg, 68%).
[0161] .sup.1H-NMR (300 MHz, DMSO-d.sub.6) .delta.: 1.3 (6H, s),
1.7 (2H, m), 2.0 (2H, m), 2.3 (6H, s), 2.7 (2H, s), 2.9 (2H, m),
3.4 (2H, m), 6.9 (1H, d, J=15.9 Hz), 7.1 (2H, s), 7.4 (1H, d,
J=15.9 Hz), 11.2 (1H, brs)
##STR00025##
[0162] To a solution of
(E)-4-amino-1-((4-(5,5-dimethyl-2,4-dioxoimidazolidin-1-yl)-2,6-dimethyls-
tyryl)sulfonyl)piperidine-4-carbonitrile (675 mg, 1.50 mmol) in
methanol (7.5 mL) and dimethylsulfoxide (0.195 mL) at room
temperature, a 2 N aqueous sodium hydroxide solution (1.6 ml, 1.6
mmol) was added and then a 30% aqueous hydrogen peroxide solution
(0.2 mL, 1.95 mmol) were slowly added dropwise. The mixture was
stirred at room temperature for one hour. Ethyl acetate, hexane,
and a saturated aqueous ammonium chloride solution were added to
the reaction mixture. The solid was collected by filtration,
washed, and dried to afford
(E)-4-amino-1-((4-(5,5-dimethyl-2,4-dioxoimidazolidine-1-yl)-2,6-dimethyl-
styryl)sulfonyl)piperidine-4-carboxamide (498 mg, 72%).
[0163] MS(ESI) m/z=464 (M+H)+
##STR00026##
[0164] A mixture of
(E)-4-amino-1-((4-(5,5-dimethyl-2,4-dioxoimidazolidine-1-yl)-2,6-dimethyl-
styryl)sulfonyl)piperidine-4-carboxamide (1.3 g, 2.8 mmol) and
palladium hydroxide on carbon (20% Pd) (wetted with approximately
50% water) (1.3 g) in methanol (21 mL) and dimethylformamide (7 mL)
was stirred under hydrogen atmosphere at room temperature for four
hours. The reaction mixture was filtered and washed, and then the
filtrate was concentrated under reduced pressure to afford
4-amino-1-((4-(5,5-dimethyl-2,4-dioxoimidazolidin-1-yl)-2,6-dimethylphene-
thyl)sulfonyl)piperidin-4-carboxamide (998 mg, 77%).
[0165] MS(ESI) m/z=466 (M+H)+
##STR00027##
[0166] To a solution of
4-amino-1-((4-(5,5-dimethyl-2,4-dioxoimidazolidin-1-yl)-2,6-dimethylphene-
thyl)sulfonyl)piperidin-4-carboxamide (120 mg, 0.258 mmol),
4-fluoro-3-(trifluoromethoxy)benzoic acid (69 mg, 0.309 mmol), and
diisopropylethylamine (0.09 ml, 0.516 mmol) in dimethylformamide
(2.5 mL),
O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosp-
hate (HATU) (118 mg, 0.309 mmol) was added. The mixture was stirred
at room temperature for 1.5 hours. The reaction mixture was
quenched with water, and then extracted with dichloromethane. The
organic layer was washed with brine, washed with anhydrous sodium
sulfate, and then concentrated under reduced pressure to afford
1-((4-(5,5-dimethyl-2,4-dioxoimidazolidin-1-yl)-2,6-dimethylphenethyl)sul-
fonyl)-4-(4-fluoro-3-(trifluoromethoxy)benzamide)piperidine-4-carboxamide
(150 mg, 67%).
[0167] MS(ESI) m/z=672 (M+H)+
##STR00028##
[0168] To a mixed solution of
1-((4-(5,5-dimethyl-2,4-dioxoimidazolidin-1-yl)-2,6-dimethylphenethyl)sul-
fonyl)-4-(4-fluoro-3-(trifluoromethoxy)benzamide)piperidine-4-carboxamide
(150 mg, 0.223 mmol) in tert-butanol (2.5 mL) and ethanol (2.5 mL),
potassium tert-butoxide (75 mg, 0.670 mmol) was added at 0.degree.
C. The mixture was stirred under nitrogen atmosphere at 50.degree.
C. for 1.5 hours. The reaction mixture was cooled, diluted with
water, quenched with a saturated aqueous ammonium chloride
solution, and then extracted with dichloromethane. The organic
layer was washed with water and brine, dried over anhydrous sodium
sulfate, and then concentrated under reduced pressure. The obtained
residue was purified by silica gel column chromatography
(dichloromethane-methanol) to afford
1-(4-(2-((2-(4-fluoro-3-(trifluoromethoxy)phenyl)-4-oxo-1,3,8-triazaspiro-
[4.5]deca-1-en-8-yl)sulfonyl)ethyl)-3,5-dimethylphenyl)-5,5-dimethylimidaz-
olidine-2,4-dione 118 mg, 81%).
[0169] MS(ESI) m/z=654 (M+H)+. .sup.1H-NMR (400 MHz, CD.sub.3OD)
.delta.: 1.40 (6H, s), 1.71-1.80 (2H, m), 2.00-2.08 (2H, m), 2.43
(6H, s), 3.22 (4H, s), 3.47-3.57 (2H, m), 3.80-3.88 (2H, m), 7.01
(2H, s), 7.50-7.57 (1H, m), 7.97-8.04 (1H, m), 8.05-8.12 (1H,
m)
[0170] The following compounds of the Examples were synthesized by
operations similar to those of Reactions 1-8 and 1-9 in Example 1,
using appropriate carboxylic acid starting materials, reagents, and
solvents.
Compound 2-5
TABLE-US-00001 [0171] TABLE 1 Com- Carboxylic acid pound starting
material Structural formula of compound Analytical data 2
##STR00029## ##STR00030## MS(ESI) m/z = 630, 632 (M + H)+.sub.o
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 1.30 (6H, s),
1.56-1.63 (2H, m), 1.80-1.90 (2H, m), 2.37 (6H, s), 3.00-3.08 (2H,
m), 3.23-3.30 (2H, m), 3.32-3.41 (2H, m), 3.67-3.73 (2H, m), 7.00
(2H, s), 7.50 (1H, dd, J = 8, 8 Hz), 7.77-7.82 7.95-8.00 (1H, m),
8.13-8.20 (1H, m), 11.10 (1H, brs), 11.70 (1H, brs) 3 ##STR00031##
##STR00032## MS(ESI) m/z = 638 (M + H)+.sub.o .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta.: 1.47 (6H, s), 1.70-1.78 (2H, m), 2.09-2.18
(2H, m), 2.40 (6H, s), 3.00-3.08 (2H, m), 3.20-3.28 (2H, m),
3.44-3.54 (2H, m), 3.80-3.88 (2H, m), 6.94 (2H, s), 7.34 (1H, t, J
= 9.6 Hz), 8.02 (1H, brs), 8.08-8.13 (1H, m), 8.20-8.24 (1H, m),
10.10 (1H, brs) 4 ##STR00033## ##STR00034## MS(ESI) m/z = 654 (M +
H)+.sub.o .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.: 1.30 (6H,
s), 1.58-1.64 (2H, m), 1.81-1.91 (2H, m), 2.37 (6H, s), 3.00-3.08
(2H, m), 3.22-3.31 (2H, m), 3.32-3.42 (2H, m), 3.68-3.73 (2H, m),
7.00 (2H, s), 7.76-7.82 (1H, m), 7.95 (1H, d, J = 9.6 Hz), 8.05
(1H, dd, J = 9.6, 2 Hz), 11.09 (1H, s), 11.79 (1H, s) 5
##STR00035## ##STR00036## MS(ESI) m/z = 632 (M + H)+.sub.o
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.47 (6H, s), 1.65-1.73
(2H, m), 2.11-2.20 (2H, m), 2.39 (6H, s), 2.98-3.04 (2H, m),
3.18-3.25 (2H, m), 3.40-3.52 (2H, m), 3.82-3.90 (2H, m), 6.94 (2H,
s), 7.17 (1H, d, J = 8.4 Hz), 7.63 (1H, d, J = 8.4 Hz), 7.75 (1H,
s), 8.49 (1H, brs), 10.46 (1H, brs)
Example 2
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazaspi-
ro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione
(Compound 6)
##STR00037##
[0173] A mixture of
2-(3-(trifluoromethyl)phenyl)-8-(vinylsulfonyl)-1,3,8-triazaspiro[4.5]dec-
a-1-en-4-one (150 mg, 0.387 mmol) synthesized according to the
method described in Schemes 2, 3, and 12 of WO2010/126030(A1),
1-(4-bromo-3,5-dimethylphenyl)-5,5-dimethylimidazolidine-2,4-dione
(169 mg, 0.542 mmol), bis(dibenzylidineacetone) palladium (45 mg,
0.077 mmol), tri-tert-butylphosphine tetrafluoroboric acid (22 mg,
0.077 mmol), and methyldicyclohexylamine (0.123 mL, 0.581 mmol) in
N-methyl-2-pyrrolidone (0.97 mL) was stirred at 100.degree. C. for
one hour under nitrogen atmosphere. The reaction mixture was
cooled, quenched with water, and then extracted with ethyl acetate.
The organic layer was washed with water and brine, dried over
anhydrous sodium sulfate, and then concentrated under reduced
pressure. The obtained residue was purified by silica gel column
chromatography (ethyl acetate-hexane) to afford
(E)-1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-tria-
zaspiro[4.5]deca-1-en-8-yl)sulfonyl)vinyl)phenyl)-5,5-dimethylimidazolidin-
e-2,4-dione (197 mg, 82%).
[0174] MS(ESI) m/z=618 (M+H)+
##STR00038##
[0175] A mixture of
(E)-1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-tria-
zaspiro[4.5]deca-1-en-8-yl)sulfonyl)vinyl)phenyl)-5,5-dimethylimidazolidin-
e-2,4-dione (195 mg, 0.316 mmol) and palladium hydroxide/carbon
(20% Pd) (wetted with approximately 50% water) (195 mg, 0.139 mmol)
in 2,2,2-trifluoroethanol (6 mL) was stirred at room temperature
for 14 hours under hydrogen atmosphere. The mixture was filtered,
and the filtrate was concentrated under reduced pressure. The
obtained residue was purified by silica gel column chromatography
(ethyl acetate-hexane) to afford
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,-
8-triazaspiro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,4-dione (121 mg,
62%).
[0176] MS(ESI) m/z=620 (M+H)+. .sup.1H-NMR (400 MHz, CD.sub.3OD)
.delta.: 1.40 (6H, s), 1.72-1.81 (2H, m), 2.00-2.10 (2H, m), 2.44
(6H, s), 3.22 (4H, s), 3.50-3.58 (2H, m), 3.80-3.88 (2H, m), 7.01
(2H, s), 7.72-7.79 (1H, m), 7.88-7.94 (1H, m), 8.16-8.23 (1H, m),
8.31 (1H, s)
Example 3
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2,-
4-dione (Compound 7)
##STR00039##
[0178] With the use of appropriate starting materials and solvents,
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-5,5-dimethylimidazolidine-2-
,4-dione (Compound 7) was synthesized by operations similar to
those described in Example 2.
[0179] MS(ESI) m/z=636 (M+H)+. .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta.: 1.47 (6H, s), 1.70-1.78 (2H, m), 2.10-2.19 (2H, m), 2.40
(6H, s), 3.00-3.07 (2H, m), 3.19-3.25 (2H, m), 3.45-3.53 (2H, m),
3.81-3.88 (2H, m), 6.94 (2H, s), 7.35 (2H, d, J=8.0 Hz), 7.73 (1H,
brs), 7.93 (2H, d, J=8.0 Hz), 9.37 (1H, brs)
Example 4
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2,-
4-dione (Compound 8)
##STR00040##
[0181] To a mixture of cyclopentanone (42 mg, 0.500 mmol) and
4-bromo-3,5-dimethylaniline (100 mg, 0.500 mmol) in acetic acid
(0.5 mL), trimethylsilyl cyanide (0.063 ml, 0.500 mmol) was added
at room temperature. The mixture was stirred at room temperature
for 1.5 hours under nitrogen atmosphere. The reaction mixture was
quenched with 28% aqueous ammonia (1 mL), diluted with water and
extracted with dichloromethane. The organic layer was washed with
water and brine, dried over anhydrous sodium sulfate, and then
concentrated under reduced pressure to afford
1-((4-bromo-3,5-dimethylphenyl)amino)cyclopentanecarbonitrile as a
crude product (152 mg).
[0182] .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.: 1.83-1.92 (4H,
m), 2.07-2.15 (2H, m), 2.33-2.42 (2H, m), 2.37 (6H, m), 3.71 (1H,
brs), 6.56 (2H, s)
##STR00041##
[0183] To a solution of
1-((4-bromo-3,5-dimethylphenyl)amino)cyclopentanecarbonitrile (145
mg, 0.495 mmol) in dichloromethane (5 mL),
2,2,2-trichloroactylisocyanate (0.070 mL, 0.593 mmol) was added at
room temperature. The mixture was stirred at room temperature for
one hour under nitrogen atmosphere.
[0184] Triethylamine (0.103 mL, 0.742 mmol), water (0.045 mL), and
methanol (0.10 mL) were added and the mixture was refluxed for 1.5
hours under nitrogen atmosphere. The reaction mixture was cooled,
followed by dilution with water and adjustment of its pH to 5 using
a 1 N aqueous hydrochloric acid solution, and then extracted with
dichloromethane. The organic layer was washed with water and brine,
dried over anhydrous sodium sulfate, and then concentrated under
reduced pressure to afford
1-(4-bromo-3,5-dimethylphenyl)-4-imino-1,3-diazaspiro[4.4]nonan-2-one
as a crude product.
[0185] MS(ESI) m/z=336, 338 (M+H)+
##STR00042##
[0186] A mixture of
1-(4-bromo-3,5-dimethylphenyl)-4-imino-1,3-diazaspiro[4.4]nonan-2-one
(the crude product obtained in the previous reaction) in acetic
acid (1.0 mL) and water (0.25 mL) was stirred for 1.5 hours at
65.degree. C. under nitrogen atmosphere. After further addition of
acetic acid (1.0 mL) and water (0.25 mL), the mixture was stirred
for 17 hours at 65.degree. C. under nitrogen atmosphere. The
reaction mixture was cooled, followed by dilution with water and
adjustment of its pH to 8 using a saturated aqueous sodium hydrogen
carbonate solution, and extracted with ethyl acetate. The organic
layer was washed with water and brine, dried over anhydrous sodium
sulfate, and then concentrated under reduced pressure. The residue
was purified by silica gel column chromatography (ethyl
acetate-hexane) to afford
1-(4-bromo-3,5-dimethylphenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione
(121 mg).
[0187] MS(ESI) m/z=337, 339 (M+H)+
##STR00043##
[0188] With the use of appropriate starting materials and solvents,
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2-
,4-dione (Compound 8) was obtained by operations similar to those
described in Example 2.
[0189] MS(ESI) m/z=662 (M+H)+. .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta.:1.36-1.44 (2H, m), 1.60-1.70 (4H. m), 1.82-1.91 (2H, m),
1.91-2.06 (4H, m), 2.38 (6H, s), 3.01-3.09 (2H, m), 3.22-3.30 (2H,
m), 3.30-3.42 (2H, m), 3.70-3.77 (2H, m), 7.03 (2H, s), 7.57 (2H,
d, J=8.4 Hz), 8.14 (2H, d, J=8.4 Hz)
Example 5
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-8-methyl-1,3,8-triazaspiro[4-
.5]decane-2,4-dione (Compound 9)
##STR00044##
[0191] With the use of 4-oxopiperidine-1-carboxylic acid tert-butyl
ester as a starting material, and the use of an appropriate
solvent,
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2,4-dioxo-1,3,8-triazaspiro-
[4.5]decan-8-carboxylic acid tert-butyl ester was obtained by
operations similar to those described in Example 4.
[0192] MS(ESI) m/z=777 (M+H)+.
##STR00045##
[0193] To a mixed solution of
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2,4-dioxo-1,3,8-triazaspiro-
[4.5]decan-8-carboxylic acid tert-butyl ester (11.7 mg, 0.015 mmol)
in dichloromethane (0.13 mL), trifluoroacetic acid (0.05 mL, 0.673
mmol) was added at room temperature. The mixture was placed under a
stream of nitrogen, and stirred at room temperature for one hour.
The reaction mixture was concentrated under reduced pressure to
obtain
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3,8-triazaspiro[4.5]decan-
-2,4-dione 2 trifluoroacetic acid salt (13.6 mg).
[0194] MS(ESI) m/z=677 (M+H)+.
##STR00046##
[0195] To a mixture of
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-1,3,8-triazaspiro[4.5]decan-
-2,4-dione 2 trifluoroacetic acid salt (21.1 mg, 0.022 mmol) and
formic acid (0.033 mL), a 37% aqueous formaldehyde solution (0.055
mL) was added. The mixture was placed under a stream of nitrogen,
and stirred for three hours while heating at 80.degree. C. The
reaction mixture was concentrated, and the resulting residue was
diluted with ethyl acetate. The organic layer was washed with a
diluted aqueous sodium hydroxide solution, dried over anhydrous
magnesium sulfate, and then concentrated under reduced pressure.
The obtained residue was subjected to column chromatography
(dichloromethane-methanol) for purification to obtain
1-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl-8-methyl-1,3,8-triazaspiro[4-
.5]decane-2,4-dione (4.5 mg, 30%).
[0196] MS(ESI) m/z=691 (M+H)+. .sup.1H-NMR (400 MHz, CD.sub.3OD)
.delta.: 1.76-1.84 (2H, m), 1.92-2.02 (2H, m), 2.02-2.12 (4H, m),
2.38 (3H, s), 2.46 (6H, s), 2.81-2.88 (2H, m), 2.92-3.02 (2H, m),
3.23 (4H, s), 3.51-3.60 (2H, m), 3.72-3.80 (2H, m), 7.01 (2H, s),
7.48 (2H, d, J=8.0 Hz), 8.10 (2H, d, J=8.0 Hz)
Example 6
5-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2-oxa-5,7-diazaspiro[3.4]oct-
ane-6,8-dione (Compound 10)
##STR00047##
[0198] With the use of oxetane-3-one as a starting material, and
the use of appropriate solvents,
5-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-2-oxa-5,7-diazaspiro[3.4]oc-
tane-6,8-dione was obtained by operations similar to those of
Example 4.
[0199] MS(ESI) m/z=650 (M+H)+. .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta.: 1.69-1.77 (2H, m), 2.12-2.22 (2H, m), 2.45 (6H, s),
3.03-3.11 (2H, m), 3.22-3.29 (2H, m), 3.46-3.53 (2H, m), 3.84-3.91
(2H, m), 4.86 (2H, d, J=7.2 Hz), 5.03 (2H, d, J=7.2 Hz), 7.07 (2H,
s), 7.35 (2H, d, J=8.4 Hz), 7.98 (2H, d, J=8.4 Hz), 8.56 (1H, s),
10.34 (1H, s)
Example 7
4-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-4,6-diazaspiro[2.4]heptane-5-
,7-dione (Compound 11)
##STR00048##
[0201] A mixture of 2-bromo-5-iodo-1,3-dimethylbenzene (300 mg,
0.965 mmol), 1-aminocyclopropane carboxylic acid (195 mg, 1.93
mmol), copper iodide (I) (37 mg, 0.194 mmol), and
diazabicycloundecene (0.50 mL, 3.35 mmol) in dimethylacetamide (2.6
mL) was stirred at 120.degree. C. for three hours under nitrogen
atmosphere. The reaction mixture was purified by silica gel column
chromatography (Wakosil C18, acetonitrile-water (0.1% formic acid))
to afford 1-((4-bromo-3,5-dimethylphenyl)amino)cyclopropane
carboxylic acid (219 mg, 80%).
[0202] MS(ESI) m/z=284, 286 (M+H)+.
##STR00049##
[0203] To a mixture of
1-((4-bromo-3,5-dimethylphenyl)amino)cyclopropane carboxylic acid
(198 mg, 0.697 mmol) in acetic acid (3 mL) and dichloromethane (1.5
mL), potassium cyanate (424 mg, 5.23 mmol) was added at room
temperature. The mixture was stirred at room temperature for one
hour, and then stirred at 60.degree. C. for two hours. A saturated
aqueous sodium hydrogen carbonate solution was added to adjust pH
to 8, and this mixture was extracted with ethyl acetate. The
organic layer was washed with water and brine, dried over anhydrous
sodium sulfate, and then concentrated under reduced pressure. The
residue was purified by silica gel column chromatography (ethyl
acetate-hexane) to afford
4-(4-bromo-3,5-dimethylphenyl)-4,6-diazaspiro[2.4]heptane-5,7-dione
(49 mg, 23%).
[0204] MS(ESI) m/z=309, 311 (M+H)+.
##STR00050##
[0205] With the use of appropriate starting materials and solvents,
4-(3,5-dimethyl-4-(2-((4-oxo-2-(4-(trifluoromethoxy)phenyl)-1,3,8-triazas-
piro[4.5]deca-1-en-8-yl)sulfonyl)ethyl)phenyl)-4,6-diazaspiro[2.4]heptane--
5,7-dione (Compound 11) was obtained by operations similar to those
of Example 2.
[0206] MS(ESI) m/z=634 (M+H)+. .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta.: 0.99-1.03 (2H, m), 1.19-1.27 (4H, m), 1.58-1.64 (2H, m),
1.81-1.90 (2H, m), 2.35 (6H, s), 2.99-3.04 (2H, m), 3.22-3.29 (2H,
m), 3.67-3.73 (2H, m), 6.95 (2H, s), 7.56 (2H, d, J=8.4 Hz), 8.12
(2H, d, J=8.4 Hz)
Example 8
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazaspi-
ro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione
(Compound 12)
##STR00051##
[0208] With the use of appropriate starting materials and solvents,
1-(3,5-dimethyl-4-(2-((4-oxo-2-(3-(trifluoromethyl)phenyl)-1,3,8-triazasp-
iro[4.5]deca-1-en-8-yl)
sulfonyl)ethyl)phenyl)-1,3-diazaspiro[4.4]nonane-2,4-dione was
obtained by operations similar to those of Example 2.
[0209] MS(ESI) m/z=646 (M+H)+. .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta.:1.40-1.48 (2H, m), 1.62-1.71 (4H, m), 1.88-1.97 (2H, m),
1.97-2.08 (4H, m), 2.41 (6H, s), 3.03-3.10 (2H, m), 2.29-3.34 (2H,
m), 3.38-3.47 (2H, m), 3.72-3.79 (2H, m), 7.06 (2H, s), 7.84 (1H,
dd, J=7.6, 7.6 Hz), 8.02 (1H, d, J=7.6 Hz), 8.33 (1H, d, J=7.6 Hz),
8.38 (1H, s)
Test Examples
[0210] For the compounds of the present invention, test results on
the activity of cAMP production via the human PTH1R, activity of
cAMP production via the rat PTH1R, metabolic stability using human
liver microsomes, metabolic stability using rat hepatocyte, and
calcemic action in TPTX rat models are shown in Test Examples 1 to
5, respectively. Compounds described in WO2010/126030A1, which are
shown in Table 2, were used as comparative compounds.
TABLE-US-00002 TABLE 2 Comparative Example Structural formula
Comparative Example 1 WO2010/126030A1 Compound 792 ##STR00052##
Comparative Example 2 WO2010/126030A1 Compound 799 ##STR00053##
Comparative Example 3 WO2010/126030A1 Compound 800 ##STR00054##
Comparative Example 4 WO2010/126030A1 Compound 878 ##STR00055##
Comparative Example 5 WO2010/126030A1 Compound 879 ##STR00056##
Comparative Example 6 WO2010/126030A1 Compound 887 ##STR00057##
Test Example 1: Measurement of In Vitro cAMP Signal Activity of
Compounds Via the Human PTH1R
(Peptides)
[0211] Human PTH (1-34) and calcitonin were purchased from Peptide
Institute, Inc. (Osaka, Japan), dissolved in 10 mM acetic acid to 1
mM and stored in a -80.degree. C. freezer.
(Cell Culture)
[0212] Cells were cultured in Dulbecco's modified Eagle's medium
(DMEM) supplemented with 10% fetal bovine serum (Hyclone), 100
units/ml penicillin G and 100 .mu.g/ml streptomycin sulfate
(Invitrogen Corp) at 37.degree. C. in a humidified atmosphere
containing 5% CO.sub.2.
[0213] cAMP signal transduction analysis utilized LLC-PK1 cells not
expressing the PTH1R, and HKRK-B7 cells, that is, LLC-PK1 cells
overexpressing the human PTH1R at 9.5.times.10.sup.5 receptors/cell
(Takasu et al., J. Bone. Miner. Res. 14:11-20, 1999).
(Camp Stimulation)
[0214] HKRK-B7 or LLC-PK1 cells were seeded into a 96-well plate at
1.times.10.sup.5 cells/well and incubated overnight. On the
following day, 50 .mu.l of cAMP assay buffer (DMEM, 2 mM IBMX, 0.2
mg/ml bovine serum albumin, 35 mM Hepes-NaOH, pH 7.4) containing
human PTH (1-34) or each compound was added and the plate was
placed in a 37.degree. C. incubator. The cells were incubated for
20 minutes. After removing the medium, the cells were washed with
100 .mu.l of cAMP assay buffer once. The plate was placed on dry
ice powder to freeze the cells and then removed from the dry ice.
The cells were lysed with 40 .mu.l of 50 mM HCl and frozen again on
dry ice. The amount of intracellular cAMP produced was measured
using a commercially available cAMP EIA kit (Biotrack cAMP EIA
system, GE health care).
(Calculation of 20% Effective Concentration (EC20) and 50%
Effective Concentration (EC50) in the Measurement of In Vitro
cAMP-Inducing Ability)
[0215] Analyses were performed using a variable gradient S-shaped
dose-response curve equation. The cAMP signaling activity of human
PTH (1-34) at 100 nM was defined as 100%, and the concentration at
which each compound shows 20% or 50% cAMP signaling activity was
calculated as EC20 or EC50.
[0216] The results obtained with HKRK-B7 cells are shown in Table
3.
[0217] The degree of cAMP response in LLC-PK1 cells was lower than
the degree in HKRK-B7 cells.
TABLE-US-00003 TABLE 3 EC20 EC50 Compound (.mu.M) (.mu.M) Compound
1 1.3 5.8 Compound 2 2.4 14 Compound 3 1.5 7.2 Compound 4 1.6 7.4
Compound 5 1.7 8.1 Compound 6 2.0 9.0 Compound 7 1.1 4.1 Compound 8
1.0 3.6 Compound 9 2.6 12 Compound 10 5.0 21 Compound 11 1.5 11
Comparative 1.5 4.8 Example 1 Comparative 3.1 13 Example 2
Comparative 2.0 9.0 Example 3 Comparative >505 >1000 Example
4 Comparative 3.1 25 Example 5 Comparative 3.6 32 Example 6
Test Example 2: Measurement of the Compounds' In Vitro cAMP
Signaling Activity Via the Rat PTH1R
[0218] Instead of HKRK-B7 cells, LLC-PK46_RATO_PTH1R cells
overexpressing rat PTH1R, which were established at Chugai
Pharmaceutical, were used to take measurements in a similar manner
to Test Example 1.
[0219] The results obtained by using LLC-PK46_RATO_PTH1R cells are
shown in Table 4.
[0220] The EC20 values of in vitro cAMP signaling activity of the
rat PTH1 receptor had a good correlation with those of human PTH1R.
A good correlation between rat and human was also seen for the EC50
values.
TABLE-US-00004 TABLE 4 EC20 EC50 Compound (.mu.M) (.mu.M) Compound
7 0.5 2.4 Compound 8 0.4 1.9 Compound 10 3.0 12 Compound 11 0.8 3.2
Comparative 0.8 2.3 Example 1
Test Example 3: Examination of Metabolic Stability Using Human
Liver Microsomes
[0221] In 0.1 M phosphate buffer (pH7.4), human liver microsomes
were incubated with a compound or a comparative example in the
coexistence of NADPH at 37.degree. C. for a specified amount of
time. The concentration of the parent compound at each reaction
time was measured using LC/MS/MS, and inherent clearance
(.mu.L/min/mg protein) was calculated from the slope of the
reaction time versus residual rate.
<Assay Conditions>
[0222] Compound concentration: 1 .mu.M Microsome: 0.5 mg/mL
NADPH: 1 mM
[0223] Reaction time: 0, 5, 15, and 30 minutes
[0224] The results are shown in Table 5. Compounds 1 to 11 showed
high metabolic stability against human liver microsomes in
comparison to Comparative Examples 1 to 6.
TABLE-US-00005 TABLE 5 Clearance Compound (.mu.l/min/mg) Compound 1
21 Compound 2 38 Compound 3 29 Compound 4 27 Compound 5 37 Compound
6 29 Compound 7 30 Compound 8 35 Compound 9 28 Compound 10 29
Compound 11 19 Compound 12 63 Comparative 84 Example 1 Comparative
61 Example 2 Comparative 74 Example 3 Comparative 74 Example 4
Comparative 112 Example 5 Comparative 154 Example 6
Test Example 4: Examination of Metabolic Stability Using Rat
Hepatocyte
[0225] Liver cells were prepared from the liver of rats (SD,
female) by a collagenase perfusion method. A compound of the
Examples or a Comparative Example was added, and this was incubated
at 37.degree. C. for a specified amount of time, followed by
addition of a reaction-stopping solution. The concentration of the
parent compound at each reaction time was measured using LC/MS/MS,
and inherent clearance (.mu.L/10.sup.6 cells/min) was calculated
from the slope of the reaction time versus residual rate.
<Assay Conditions>
[0226] Cell concentration: 1.times.10.sup.6 cells/mL Compound
concentration: 1 .mu.M Medium: Williams' medium E Reaction time: 0,
15, 30, 60, 120, and 240 minutes Reaction-stopping solution:
acetonitrile/2-propanol (4/6, v/v)
[0227] The results are shown in Table 6. The rat hepatocyte
metabolic stability of Compounds 2, 4, 5, 7, 8, 9, 10, and 11
increased compared to Comparative Examples 1, 2, 3, 5, and 6.
TABLE-US-00006 TABLE 6 Clearance Compound (.mu.L/10.sup.6
cells/min) Compound 1 7.6 Compound 2 3.0 Compound 3 17 Compound 4
2.2 Compound 5 1.0 Compound 6 1.4 Compound 7 0.9 Compound 8 3.0
Compound 9 1.8 Compound 10 0.3 Compound 11 -0.6 Comparative 5.8
Example 1 Comparative 5.9 Example 2 Comparative 22 Example 3
Comparative 22 Example 5 Comparative 22 Example 6
Test Example 5: Calcemic Action in the TPTX Rat Model
[0228] Four-week old female Crl:CD(SD) rats were obtained from
Charles River Japan (Atsugi Breeding Center), and were acclimated
to standard laboratory conditions of 20-26.degree. C. and 35-75%
humidity for one week. The rats were given tap water and were fed
ad libitum with standard rodent chow (CE-2) (CLEA Japan, Inc.)
containing 1.1% calcium, 1.0% phosphoric acid, and 250 IU/100 g of
vitamin D3.
[0229] TPTX was performed on five-week old rats. Some of the
individuals were subjected to sham operation (Sham). Individuals
whose serum Ca concentration was less than 8 mg/dL on four days
after the operation were selected for use as TPTX rats. On five
days after the operation, the rats were assigned to eight TPTX
groups and one Sham group (n=5, each group) based on their body
weight and serum Ca concentration measured on four days after the
operation. The solvent alone was orally administered to the Sham
group and the TPTX-Vehicle group at a volume of 10 mL/kg. Each test
article was orally administered individually to each TPTX test
article group by dissolving it in a solvent at a dose of 30 mg/10
mL/kg. The solvent composition was 10% dimethylsulfoxide (Wako Pure
Chemical Industries, Ltd.), 10% Cremophor EL (Sigma-Aldrich Japan
LLC), 20% hydroxypropyl-.beta.-cyclodextrin (Nihon Shokuhin Kako
Co., Ltd.), glycine (Wako Pure Chemical Industries, Ltd.); and the
pH was adjusted to 10. Immediately before administration of each
sample, Pre-blood collection was performed, and blood collection
was carried out at 2, 6, 10, and 24 hours after administration to
measure the serum Ca concentration. Each blood collection was
carried out from the jugular vein under isoflurane inhalation
anesthesia.
[0230] Serum Ca measurement: Serum obtained by centrifugation from
the collected blood was measured by using an automatic analyzer
TBA-120FR (Toshiba Medical Systems Corporation).
[0231] For statistical analysis of the animal studies, data are
shown as mean.+-.standard error (SE). Statistical analysis were
performed by unpaired test of the SAS Preclinical Package
(Ver.5.00.010720, SAS Institute Japan, Tokyo, Japan). A p-value of
<0.05 was regarded as statistically significant. Statistically
significant of each test article group comparing to the
TPTX-Vehicle group, the Comparative Example 1 group, and the
Comparative Example 2 group was shown as #, *, and .intg.
respectively.
[0232] The Pre-value for the serum Ca concentration was 9.9 mg/dL
for the Sham group, and 5.3-6.2 mg/dL for each of the TPTX groups.
The serum Ca concentrations for each compound up to 24 hours after
administration are shown in the FIGURE as the average amount of
change from the Pre-value. Furthermore, for all of the compounds,
the serum Ca concentration peaked at six hours after administration
or ten hours after administration of each compound.
[0233] Compounds 6, 7, and 8 which have high rat hepatocyte
metabolic stability showed large positive changes from the
Pre-value, and their oral administration showed strong effects on
calcemic action. On the other hand, Compound 1, and Comparative
Examples 1 and 2 which have low rat hepatocyte metabolic stability
showed smaller positive changes from the Pre-value compared to
Compounds 6, 7, and 8. In particular, Compounds 7 and 8 were
statistically significant compared to Comparative Examples 1 and
2.
[0234] Furthermore, Compounds 6, 7, and 8 which have high rat
hepatocyte metabolic stability showed individual maximum values of
7.8 to 8.5 mg/dL at six or ten hours after administration, and
achieved the therapeutic target range of serum Ca concentration of
7.6 to 8.8 mg/dL in hypoparathyroidism patients. On the other hand,
this therapeutic target range could not be achieved at any of the
measurement times for Compound 1, and Comparative Examples 1 and 2
which have low rat hepatocyte metabolic stability.
[0235] From the above-mentioned test results, Compounds 6, 7, and
8, which have strong cAMP-signaling activities in cells forced to
express rat PTH1R and high stability against metabolic breakdown in
rat hepatocytes were found to show strong effects on calcemic
action in rats when administered orally. These compounds also have
cAMP-signaling activity in cells forced to express human PTH1R and
high metabolic stability against human liver microsomes compared to
the Comparative Compounds; and they are expected to have high
therapeutic effects when administered orally to hypoparathyroidism
patients. Furthermore, compounds represented by Formula (1), which
have cAMP-signaling activity in cells forced to express human PTH1R
and show metabolic stability against human liver microsomes to the
same degree as Compounds 6, 7, and 8, are also expected to have
high therapeutic effects in hypoparathyroidism patients.
INDUSTRIAL APPLICABILITY
[0236] The present invention provides compounds having a strong
PTH-like effect and high metabolic stability. The present invention
also provides a medicine for the prevention and/or treatment of
osteoporosis, fracture, adynamic bone disease, achondronplasia,
hypochondroplasia, osteomalacia, osteoarthritis, arthritis,
thrombocytopenia, hypoparathyroidism, hyperphosphatemia, tumoral
calcinosis or the like, or stem cell mobilization.
* * * * *