Note: Descriptions are shown in the official language in which they were submitted.
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UREA DERIVATIVES AS. ANTAGONISTS OF THE VANILLOID RECEPTOR (VRl)
DETAILED DESCRIPTION OF INVENTION
TECHNICAL FIELD
The present invention relates to a urea derivative which is useful as an
active ingredient of
pharmaceutical preparations. The urea derivative of the present invention has
vanilloid receptor
(VRl) antagonistic activity, and can be used for the prophylaxis and treatment
of diseases
associated with VRl activity, in particular for the treatment of urological
diseases or disorders,
such as detrusor overactivity (overactive bladder), urinary incontinence,
neurogenic detrusor
overactivity (detrusor hyperflexia), idiopathic detrusor overactivity
(detrusor instability), benign
prostatic hyperplasia, and lower urinary tract symptoms; chronic pain,
neuropathic pain,
postoperative pain, rheumatoid arthritic pain, neuralgia, neuropathies,
algesia, nerve injury,
ischaemia, neurodegeneration, stroke; and respiratory diseases and
inflammatory disorders such as
asthma, chronic obstructive pulmonary (or airways) disease (COPD), common
cold, cough, sneeze,
bronchitis including acute and chronic bronchitis, bronchiolitis, rhinitis,
allergic rhinitis,
vasomotor rhinitis, mucositis, sinusitis, allergy, disorders associated with
exogenous irritants such
as tobacco smoke, smog, high levels of atmospheric SOZ and noxious gases in
the workplace, and
airways hyperreactivity, milk product intolerance, Loffler's pneumonia,
emphysema, cystic
fibrosis, bronchiectasis, pulmonary fibrosis, pneumoconiosis, collagen
vascular disease,
granulomatous disease, laryngitis, pharyngitis, pneumonia, pleuritis,
persistent asthma and chronic
asthmatic bronchitis.
BACKGROUND ART
Vanilloid compounds are characterized by the presence of vanillyl group or a
functionally
equivalent group. Examples of several vanilloid compounds or vanilloid
receptor modulators are
vanillin (4-hydroxy-3-methoxy-benzaldehyde), guaiacol (2-methoxy-phenol),
zingerone (4-/4-hy-
droxy-3-methoxyphenyl/-2-butanon), eugenol(2-methoxy4-/2-propenyl/phenol), and
capsaicin
(8-methy-N-vanillyl-6-noneneamide).
Among others, capsaicin, the main pungent ingredient in "hot" chili peppers,
is a specific
neurotoxin that desensitizes C-fiber afferent neurons. Capsaicin interacts
with vanilloid receptors
(VRl), which are predominantly expressed in cell bodies of dorsal root ganglia
(DRG) or nerve
endings of afferent sensory fibers including C-fiber nerve endings [Tominaga
M, Caterina MJ,
Malmberg AB, Rosen TA, Gilbert H, Skinner K, Raumann BE, Basbaum AI, Julius D:
The cloned
capsaicin receptor integrates multiple pain-producing stimuli. Neuron. 21: 531-
543, 1998]. The
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VR1 receptor was recently cloned [Caterina MJ, Schumacher MA, Tominaga M,
Rosen TA,
Levine JD, Julius D: Nature 389: 816-824, (1997)] and identified as a
nonselective cation channel
with six transmembrane domains that is structurally related to the TRP
(transient receptor
potential) channel family. Binding of capsaicin to VRl allows sodium, calcium
and possibly
potassium ions to flow down their concentration gradients, causing initial
depolarization and
release of neurotransmitters from the nerve terminals. VRl can therefore be
viewed as a molecular
integrator of chemical and physical stimuli that elicit neuronal signals in
pathological conditions or
diseases.
There is abundant direct or indirect evidence that shows the relation between
VRl activity and
diseases such as pain, ischaemia, and inflammatory disorders (e.g., WO
99/00115 and 00/50387).
Further, it has been demonstrated that VRl transduces reflex signals that are
involved in the
overactive bladder of patients who have damaged or abnormal spinal reflex
pathways [De Groat
WC: A neurologic basis for the overactive bladder. Urology 50 (6A Supply: 36-
52, 1997].
Desensitisation of the afferent nerves by depleting neurotransmitters using
VRl agonists such as
capsaicin has been shown to give promising results in the treatment of bladder
dysfunction
associated with spinal cord injury and multiple sclerosis [(Maggi CA:
Therapeutic potential of
capsaicin-like molecules - Studies in animals and humans. Life Sciences 51:
1777-1781, 1992) and
(DeRidder D; Chandiramani V; Dasgupta P; VanPoppel H; Baert L; Fowler CJ:
Intravesical
capsaicin as a treatment for refractory detrusor hyperreflexia: A dual center
study with long-term
followup. J. Urol. 158: 2087-2092, 1997)].
It is anticipated that antagonism of the VRl receptor would lead to the
blockage of
neurotransmitter release, resulting in prophylaxis and treatment of the
conditions and diseases
associated with VRl activity.
It is therefore expected that antagonists of the VR1 receptor can be used for
prophylaxis and
treatment of the conditions and diseases including chronic pain, neuropathic
pain, postoperative
pain, rheumatoid arthritic pain, neuralgia, neuropathies, algesia, nerve
injury, ischaemia,
neurodegeneration, stroke, inflammatory disorders, urinary incontinence (UI)
such as urge urinary
incontinence (UUI), and/or overactive bladder.
UI is the involuntary loss of urine. UUI is one of the most common types of UI
together with stress
urinary incontinence (SUI) which is usually caused by a defect in the urethral
closure mechanism.
UUI is often associated with neurological disorders or diseases causing
neuronal damages such as
dementia, Parkinson's disease, multiple sclerosis, stroke and diabetes,
although it also occurs in
individuals with no such disorders. One of the usual causes of UUI is
overactive bladder (OAB)
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which is a medical condition referring to the symptoms of frequency and
urgency derived from
abnormal contractions and instability of the detrusor muscle.
There are several medications for urinary incontinence on the market today
mainly to help treating
UUI. Therapy for OAB is focused on drugs that affect peripheral neural control
mechanisms or
those that act directly on bladder detrusor smooth muscle contraction, with a
major emphasis on
development of anticholinergic agents. These agents can inhibit the
parasympathetic nerves which
control bladder voiding or can exert a direct spasmolytic effect on the
detrusor muscle of the
bladder. This results in a decrease in intravesicular pressure, an increase in
capacity and a
reduction in the frequency of bladder contraction. Orally active
anticholinergic drugs which are
commonly prescribed have serious drawbacks such as unacceptable side effects
such as dry mouth,
abnormal visions, constipation, and central nervous system disturbances. These
side effects lead to
poor compliance. Dry mouth symptoms alone are responsible for a 70% non-
compliance rate with
oxybutynin. The inadequacies of present therapies highlight the need for
novel, efficacious, safe,
orally available drugs that have fewer side effects.
W003/014064 discloses the compounds represented by the general formula:
Rb\NiX
Raa
\Qaa~
Y
wherein
X represents C3_8 cycloalkyl optionally fused by benzene, optionally
substituted naphthyl,
optionally substituted phenyl, optionally substituted phenyl C1_6 straight
alkyl, phenyl
fused by cycloalykyl, etc;
Qaa represents CH or N;
Raa represents hydrogen or methyl;
Rbb represents hydrogen or methyl; and
Y represents substituted naphthyl,
as a vanilloid receptor antagonist.
W003/022809 discloses the compounds having vanilloid receptor antagonist
activity represented
by the general formula:
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(Ra2)9
H H
N\ /N
\ (CHz)n
P O
(Rat)P
wherein
P and P' independently represent aryl or heteroaryl;
Ray and Raa independently represent hydrogen, alkoxy, hydroxy, etc;
n is 0, 1, 2 or 3; p and q are independently 0,1, 2, 3 or 4; r is 1, 2 or 3;
and s is 0, 1 or 2.
W003/053945 discloses the compounds having vanilloid receptor antagonist
activity represented
by the general formula:
N N Rbz
\ (CHz)~/
Pa
O
(Rb1)P
wherein
Pe represents phenyl, naphthyl or heterocyclyl;
n is 2, 3, 4, 5 or 6; p is independently 0,1, 2, 3 or 4;
Rb' represents hydrogen, alkoxy, hydroxy, etc; and
Raz represents
(Rbs)
9
~i''N
~X
( Rb4) f
1 S wherein X is a bond, C, O, or NRbB; and r, q, R63, Rb4 are defined in the
application.
W003/070247 discloses the compounds having vanilloid receptor antagonist
activity represented
by the general formula:
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R'~a Zc~
c
~ c9
Xc_ Zc" R
Xc ~c~
z
Xc~~.
7Cc4
R~
wherein
Xcl represents N or CR°I; Xcz represents N or CR°z; Xc3
represents N, NRc3 or CR°3;
Xc4 represents a bond, N or CRS; Xcs represents N or C; provided that at least
one of Xcl, Xcz,
Xc3 and Xc4 is N; Zcl represents O, NH or S; Zcz represents a bond, NH or S;
L° represents
alkylene, cycloallcylene, etc; Rc', Rcz, R°3, R°a, R~s, Rcb,
R°', Rcsa Rcsb are defined in the application;
and R°9 represents hydrogen, aryl, cycloalkyl, and heterocylcle.
W003/080578 discloses the compounds having vanilloid receptor antagonist
activity represented
by the general formula:
~Rdt~~_3 Xd
~ /~CRd5Rd6~n\ld
N' _N
\ ( d3 ~ d4
(Rd2~~_3 R R
d d
B\Dd:E
wherein
Aa, Ba, Da and Ea are each C or N with the proviso that one or more are N; Xa
is an O, S or =NCN;
Ya is an aryl, heteroaryl, carbocyclyl or fused-carbocyclyl; n is 0, l, 2 or
3; and Ral, Raz, Rd3~ Raa
Ras and Ra6 are defined in the application.
The development of a compound which has effective VRl antagonistic activity
and can be used for
the prophylaxis and treatment of diseases associated with VR1 activity, in
particular for the
treatment of urinary incontinence, urge urinary incontinence, overactive
bladder as well as pain,
and/or inflammatory diseases such as asthma and COPD has been desired.
SUMMARY OF THE INVENTION
This invention is to provide a urea derivatives of the formula ()7, their
tautomeric and
stereoisomeric form, and salts thereof
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1
X ~ ~R
N_ _N' L Jri N-R2 (>]
H H
O N
4
R3 ~ R )1-3
wherein
n represents 0, 1, 2, 3, or 4;
R' represents 3-8 membered saturated or unsaturated ring optionally having one
to
three hetero atoms selected from the group consisting of N, O, and S
wherein
said ring is optionally substituted with one or more substituents
independently
selected from the group consisting of halogen, nitro, thiol, hydroxy,
carboxy, cyano, amino, N-(C~_balkyl)amino, N,N-di(CI_6alkyl)amino, N-
(C3_g cycloalkyl)amino, C~_6alkoxycarbonyl, sulfonamide, C~_6 alkanoyl, N-
(C1_6alkanoyl)amino, carbamoyl, CI_6 alkylcarbamoyl, C3_$cycloalkyl, hete-
rocycle,
C,_6 alkyl [wherein said alkyl is optionally substituted by cyano, nitro,
hydroxy, carboxy, amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-
halogen], CZ_6 alkenyl [wherein said alkenyl is optionally substituted by
cyano, nitro, hydroxy, carboxy, amino, C,_6 alkoxycarbonyl or mono-, di-,
or tri-halogen], C1_6 alkoxy [wherein said alkoxy is optionally substituted by
mono-, di-, or tri- halogen], C~_6 alkylthio [wherein said alkylthio is
optionally substituted by mono-, di-, or tri- halogen], phenyl, benzyl and
phenoxy [wherein said phenyl, phenyl moiety of said benzyl or phenyl
moiety of said phenoxy are optionally substituted by halogen, nitro,
hydroxy, carboxy, amino, N-(Cl_6alkyl)amino, mercapto, C~_6 alkylthio, C~_6
alkylsulfinyl, Cl_6 alkylsulfonyl, N,N-di(CI_6 alkyl)amino, N-(C3_$
cycloalkyl)amino, C~_6 alkoxycarbonyl, C1_6 alkoxycarbonyl or C~_6 alkyl];
RZ represent hydrogen, C~_6alkyl, CZ_6alkenyl, CZ_6alkynyl, CI_6alkylsulfonyl,
C~_balkyloxycarbonyl, C~_6alkylcarbonyl, or C3_8cycloalkyl
wherein
said alkyl, alkenyl or alkynyl are optionally substituted by mono-, di-, or
tri-
halogen, hydroxy, carboxyl, nitro, cyano, Cl_6alkylcarbonyl, C~_6alkoxy,
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C,_balkoxycarbonyl, C3_$cycloalkyl, amino, N-(C~_balkyl)amino, N,N-
di(C1_balkyl)amino, N-(aryl)amino, N-(heteroaryl)amino, carbamoyl,
N-(C~_6alkyl)aminocarbonyl, or N,N,-di(C~_6alkyl)aminocarbonyl,
or said cycloalkyl are optionally substituted with one or more substituents
independently selected from the group consisting of halogen, hydroxy,
carboxyl,
cyano, nitro, (C1_balkoxy)carbonyl, C3_8cycloalkyl, amino, N-(C~_6alkyl)amino,
N,N-di(C~_6alkyl)amino, N-(aryl)amino, N-(heteroaryl)amino, carbamoyl, N-
(C~_6alkyl)aminocarbonyl, N,N-di(C~_balkyl)aminocarbonyl, C~_6alkyl optionally
substituted by mono-, di-, or tri-halogen, or CI_6alkoxy optionally
substituted by
mono-, di-, or tri-halogen; or
R' and R2 together with the adjacent N form S-12 membered saturated or
unsautrated
cyclic ring optionally having one N or O atom other than the adjacent N and
said ring is
optionally substituted with one or more substituents independently selected
from the group
consisting of
halogen, nitro, thiol, hydroxy, carboxy, cyano, amino, N-(Cl_6alkyl)amino, N,N-
di(C~_6alkyl)amino, N-(C3_$ cycloalkyl)amino, CI_6alkoxycarbonyl, sulfonamide,
Cl_6 alkanoyl, N-(C~_6alkanoyl)amino, carbamoyl, C1_6 alkylcarbamoyl,
C3_8cyclo-
alkyl, heterocycle,
C1_6 alkyl [wherein said alkyl is optionally substituted by cyano, nitro,
hydroxy,
carboxy, amino, CI_6 alkoxycarbonyl or mono-, di-, or tri-halogen], CZ_6
alkenyl
[wherein said alkenyl is optionally substituted by cyano, nitro, hydroxy,
carboxy,
amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen], C~_6 alkoxy
[wherein
said alkoxy is optionally substituted by mono-, di-, or tri- halogen], C~_6
alkylthio
[wherein said alkylthio is optionally substituted by mono-, di-, or tri-
halogen],
phenyl, benzyl and phenoxy [wherein said phenyl, phenyl moiety of said benzyl
or
phenyl moiety of said phenoxy are optionally substituted by halogen, nitro,
hydroxy, carboxy, amino, N-(C~_6alkyl)amino, mercapto, C1_6 alkylthio, C~_6
alkylsulfinyl, C1_6 alkylsulfonyl, N,N-di(C~_6 alkyl)amino, N-(C3_$ cycloal-
kyl)amino, Cl_6 alkoxycarbonyl, C~_6 alkoxycarbonyl or C~_6 alkyl]
R3 represents hydrogen, C~_balkyl, CZ_balkenyl, or Ca_6alkynyl;
R4 each independently represents halogen, nitro, thiol, hydroxy, carboxy,
cyano, amino, N-
(C,_6alkyl)amino, N,N-di(C~_6alkyl)amino, N-(C3_$ cycloalkyl)amino,
C~_6alkoxycarbonyl,
sulfonamide, C~_6 alkanoyl, N-(CI_6alkanoyl)amino, carbamoyl, C,_6
alkylcarbamoyl,
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C3_8cycloalkyl, C1_6 alkyl [wherein said alkyl is optionally substituted by
cyano, nitro,
hydroxy, carboxy, amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen],
CZ_6 alkenyl
[wherein said alkenyl is optionally substituted by cyano, nitro, hydroxy,
carboxy, amino,
C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen], CI_6 alkoxy [wherein said
alkoxy is
optionally substituted by mono-, di-, or tri- halogen], or C~_6 alkylthio
[wherein said
alkylthio is optionally substituted by mono-, di-, or tri- halogen]; and
X represents O, CH2, S, or N(Rlo),
wherein
R'° represents hydrogen or C~_6 alkyl.
In another embodiment, the urea derivatives of formula (I) can be those
wherein;
n represents 0, l, 2, or 3;
R' represents phenyl, pyridine, or pyrimidine
wherein
said phenyl, pyridine, or pyrimidine is optionally substituted with one or
more
substituents independently selected from the group consisting of halogen,
nitro, thiol, hydroxy, carboxy, cyano, amino, N-(C~_6alkyl)amino, N,N-
di(CI_6alkyl)amino, N-(C3_$ cycloalkyl)amino, C~_6alkoxycarbonyl, sulfon-
amide, CI_6 alkanoyl, N-(CI_6alkanoyl)amino, carbamoyl, CI_6 alkylcarb-
amoyl, C3_8cycloalkyl, heterocycle,
C~_6 alkyl [wherein said alkyl is optionally substituted by cyano, nitro,
hydroxy, carboxy, amino, CI_6 alkoxycarbonyl or mono-, di-, or tri-
halogen], Cz_6 alkenyl [wherein said alkenyl is optionally substituted by
cyano, nitro, hydroxy, carboxy, amino, C1_6 alkoxycarbonyl or mono-, di-,
or tri-halogen], C1_6 alkoxy [wherein said alkoxy is optionally substituted
by mono-, di-, or tri- halogen], C~_6 alkylthio [wherein said alkylthio is
optionally substituted by mono-, di-, or tri- halogen], phenyl, benzyl and
phenoxy [wherein said phenyl, phenyl moiety of said benzyl or phenyl
moiety of said phenoxy are optionally substituted by halogen, nitro,
hydroxy, carboxy, amino, N-(C~_6alkyl)amino, mercapto, C1_6 alkylthio, C1_6
alkylsulfinyl, C~_6 alkylsulfonyl, N,N-di(C~_6 alkyl)amino, N-(C3_$
cycloalkyl)amino, Cl_6 alkoxycarbonyl, C1_6 alkoxycarbonyl or C~_6 alkyl];
and
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Ra represent hydrogen, C1_balkyl, Cz_6alkenyl, CZ_balkynyl, CI_6alkylsulfonyl,
or C3_
$cycloalkyl
wherein
said alkyl, alkenyl or alkynyl are optionally substituted by mono-, di-, or
tri-
halogen, hydroxy, carboxyl, nitro, cyano, C~_6alkylcarbonyl, Cl.6alkoxy,
C~_6alkoxycarbonyl, C3_8cycloalkyl, amino, N-(Cl.6alkyl)amino, N,N-
di(C~.balkyl)amino, N-(aryl)amino, N-(heteroaryl)amino, carbamoyl, N-
(C~_6alkyl)arninocarbonyl, or N,N,-di(C~_6alkyl)aminocarbonyl,
or said cycloalkyl are optionally substituted with one or more substituents
independently selected from the group consisting of halogen, hydroxy,
carboxyl,
cyano, nitro, (C~_6alkoxy)carbonyl, C3_gcycloalkyl, amino, N-(C~_balkyl)amino,
N,N-di(C,_6alkyl)amino, N-(aryl)amino, N-(heteroaryl)amino, carbamoyl, N-
(C~.balkyl)aminocarbonyl, N,N-di(Cl_6alkyl)aminocarbonyl, C~_6alkyl optionally
substituted by mono-, di-, or tri-halogen, or C~_6alkoxy optionally
substituted by
mono-, di-, or tri-halogen; or
R' and RZ together with the adjacent N form 5-12 membered saturated or
unsautrated
cyclic ring optionally having one N or O atom other than the adjacent N and
said ring is
optionally substituted with one or more substituents independently selected
from the group
consisting of
halogen, nitro, thiol, hydroxy, carboxy, cyano, amino, N-(C~_6alkyl)amino, N,N-
di(C~_6alkyl)amino, N-(C3.$ cycloalkyl)amino, C,.balkoxycarbonyl, sulfonamide,
CI_6 alkanoyl, N-(C1_6alkanoyl)amino, carbamoyl, C~_6 alkylcarbamoyl, C3_8cyc-
loalkyl, heterocycle,
C~_6 alkyl [wherein said alkyl is optionally substituted by cyano, nitro,
hydroxy,
carboxy, amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen], C~.6
alkoxy
[wherein said alkoxy is optionally substituted by mono-, di-, or tri-
halogen], CI_6
alkylthio [wherein said alkylthio is optionally substituted by mono-, di-, or
tri-
halogen], phenyl, benzyl and phenoxy [wherein said phenyl, phenyl moiety of
said
benzyl or phenyl moiety of said phenoxy are optionally substituted by halogen,
nitro, hydroxy, carboxy, amino, N-(Cl.6alkyl)amino, mercapto, C~_6 alkylthio,
C~_6
alkylsulfinyl, C~_6 alkylsulfonyl, N,N-di(C~_6 alkyl)amino, N-(C3_g cyclo-
alkyl)amino, C1_6 alkoxycarbonyl, C,_6 alkoxycarbonyl or CI_6 alkyl]
R3 represents hydrogen;
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R4 each independently represents halogen, nitro, thiol, hydroxy, carboxy,
cyano, amino, N-
(Cl_6alkyl)amino, N,N-di(C1_6alkyl)amino, N-(C3_$ cycloalkyl)amino,
C1_balkoxycarbonyl,
sulfonamide, Cl_6 alkanoyl, N-(CI_6alkanoyl)amino, carbamoyl, Cl_6
alkylcarbamoyl,
C3_$cycloalkyl, C~_6 alkyl [wherein said alkyl is optionally substituted by
cyano, nitro,
hydroxy, carboxy, amino, CI_6 alkoxycarbonyl or mono-, di-, or tri-halogen],
CZ_6 alkenyl
[wherein said alkenyl is optionally substituted by cyano, nitro, hydroxy,
carboxy, amino,
C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen], C1_6 alkoxy [wherein said
alkoxy is
optionally substituted by mono-, di-, or tri- halogen], or CI_6 alkylthio
[wherein said
alkylthio is optionally substituted by mono-, di-, or tri- halogen]; and
X represents O, CHz, S, or N(R'°),
wherein
R'° represents hydrogen or Cl_6 alkyl.
In another embodiment, the urea derivative of formula ()7 are those wherein;
n represents 0, 1, 2, or 3;
R' represents 3-8 membered saturated or unsaturated ring optionally having one
to
three hetero atoms selected from the group consisting of N, O, and S
wherein
said ring is optionally substituted with one or more substituents
independently
selected from the group consisting of halogen, nitro, thiol, hydroxy,
carboxy, cyano, amino, N-(Cl_6alkyl)amino, N,N-di(C1_balkyl)amino,
N-(C3_$ cycloalkyl)amino, C~_6alkoxycarbonyl, sulfonamide, C~_6 alkanoyl,
N-(CI_6alkanoyl)amino, carbamoyl, C~_6 alkylcarbamoyl, C3_8cycloalkyl,
heterocycle,
C~_6 alkyl [wherein said alkyl is optionally substituted by cyano, nitro,
hydroxy, carboxy, amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-
halogen], C~_6 alkoxy [wherein said alkoxy is optionally substituted by
mono-, di-, or tri- halogen], C,_6 alkylthio [wherein said alkylthio is
optionally substituted by mono-, di-, or tri- halogen], phenyl, benzyl and
phenoxy ,
[wherein said phenyl, phenyl moiety of said benzyl or phenyl moiety of
said phenoxy are optionally substituted by halogen, nitro, hydroxy, carb-
oxy, amino, N-(CI_6alkyl)amino, mercapto, Cl_6 alkylthio, CI_6 alkyl-
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sulfinyl, CI_6 alkylsulfonyl, N,N-di(CI_6 alkyl)amino, N-(C3_$ cyclo-
alkyl)amino, CI_6 alkoxycarbonyl, C1_6 alkoxycarbonyl or C~_6 alkyl]; and
Rz represent hydrogen or C,_6alkyl,
wherein
said alkyl is optionally substituted by mono-, di-, or tri-halogen, hydroxy,
carboxyl, nitro, cyano, CI_balkylcarbonyl, C~.balkoxy, Cl_6alkoxycarbonyl,
C3_$cycloalkyl, amino, N-(CI_6alkyl)amino, N,N-di(C~_6alkyl)amino, N-
(aryl)amino, N-(heteroaryl)amino, carbamoyl, N-(C~_6alkyl)aminocarbonyl,
or N,N,-di(C~_6alkyl)aminocarbonyl,
R3 represents hydrogen;
R4 each independently represents halogen, nitro, thiol, hydroxy, carboxy,
cyano, amino, N-
(C~_6alkyl)amino, N,N-di(Cl_6alkyl)amino, N-(C3_8 cycloalkyl)amino,
C,_6alkoxycarbonyl,
sulfonamide, C~.6 alkanoyl, N-(Cl.6alkanoyl)amino, carbamoyl, C,_6
alkylcarbamoyl,
C3_8 cycloalkyl, C~.6 alkyl [wherein said alkyl is optionally substituted by
cyano, nitro,
hydroxy, carboxy, amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen],
CZ_6 alkenyl
[wherein said alkenyl is optionally substituted by cyano, nitro, hydroxy,
carboxy, amino,
CI_6 alkoxycarbonyl or mono-, di-, or tri-halogen], Cl_6 alkoxy [wherein said
alkoxy is
optionally substituted by mono-, di-, or tri- halogen], or C~.6 alkylthio
[wherein said
alkylthio is optionally substituted by mono-, di-, or tri- halogen]; and
X represents O, CH2, S, or N(Rlo),
wherein
R'° represents hydrogen or C~_6 alkyl.
Yet in another embodiment, the urea derivative of formula (I) are those
wherein;
n represents 1 or 2;
Rl represents phenyl, pyridine, or pyrimidine
wherein
said phenyl, pyridine, or pyrimidine is optionally substituted with one or
more
substituents independently selected from the group consisting of halogen,
nitro, thiol, hydroxy, carboxy, cyano, amino, N-(C~.6alkyl)amino, N,N-
di(C1_balkyl)amino, N-(C3_8 cycloalkyl)amino, Cl_~alkoxycarbonyl, sulfon-
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amide, C~_6 alkanoyl, N-(CI_6alkanoyl)amino, carbamoyl, CI_6 alkylcarb-
amoyl, C3_$cycloalkyl, heterocycle,
C~_6 alkyl [wherein said alkyl is optionally substituted by cyano, nitro,
hydroxy, carboxy, amino, Cl_6 alkoxycarbonyl or mono-, di-, or tri-
halogen], C~_6 alkoxy [wherein said alkoxy is optionally substituted by
mono-, di-, or tri- halogen], C~_6 alkylthio [wherein said alkylthio is
optionally substituted by mono-, di-, or tri- halogen], phenyl, benzyl and
phenoxy ,
[wherein said phenyl, phenyl moiety of said benzyl or phenyl moiety of said
phenoxy are optionally substituted by halogen, nitro, hydroxy, carboxy,
amino, N-(CI_6alkyl)amino, mercapto, C1_6 alkylthio, CI_6 alkylsulfinyl, C~_6
alkylsulfonyl, N,N-di(C1_6 alkyl)amino, N-(C3_$ cycloalkyl)amino, C,_6
alkoxycarbonyl, C~_6 alkoxycarbonyl or C~_6 alkyl]; and
RZ represent hydrogen, C~_6alkyl, CZ_6alkenyl, CZ_6alkynyl, C,_6alkylsulfonyl,
or
C3_$ cycloalkyl
wherein
said alkyl, alkenyl or alkynyl are optionally substituted by mono-, di-, or
tri-
halogen, hydroxy, carboxyl, nitro, cyano, C1_6alkylcarbonyl, C~_balkoxy,
C1_6alkoxycarbonyl, C3_8cycloalkyl, amino, N-(CI_6alkyl)amino, N,N-
di(C~_6alkyl)amino, N-(aryl)amino, N-(heteroaryl)amino, carbamoyl, N-
(C~.6alkyl)aminocarbonyl, or N,N,-di(C~_6alkyl)aminocarbonyl,
or said cycloalkyl are optionally substituted with one or more substituents
independently selected from the group consisting of halogen, hydroxy,
carboxyl,
cyano, nitro, (C~_balkoxy)carbonyl, C3_8cycloalkyl, amino, N-(C1_balkyl)amino,
N,N-di(C,_6alkyl)amino, N-(aryl)amino, N-(heteroaryl)amino, carbamoyl,
N-(C1_6alkyl)aminocarbonyl, N,N-di(C~_balkyl)aminocarbonyl, C~_6alkyl
optionally
substituted by mono-, di-, or tri-halogen, or C~_6alkoxy optionally
substituted by
mono-, di-, or tri-halogen;
R3 represents hydrogen;
R4 each independently represents halogen, nitro, thiol, hydroxy, carboxy,
cyano, amino, N-
(CI_6alkyl)amino, N,N-di(C~_6alkyl)amino, N-(C3_8 cycloalkyl)amino,
Cl_6alkoxycarbonyl,
sulfonamide, C1_6 alkanoyl, N-(CI_6alkanoyl)amino, carbamoyl, CI_6
alkylcarbamoyl,
C3_8cycloalkyl, C1_6 alkyl [wherein said alkyl is optionally substituted by
cyano, nitro,
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hydroxy, carboxy, amino, C~_6 alkoxycarbonyl or mono-, di-, or tri-halogen],
CZ_6 alkenyl
[wherein said alkenyl is optionally substituted by cyano, nitro, hydroxy,
carboxy, amino,
C,_6 alkoxycarbonyl or mono-, di-, or tri-halogen], C,_6 alkoxy [wherein said
alkoxy is
optionally substituted by mono-, di-, or tri- halogen], or CI_6 alkylthio
[wherein said
alkylthio is optionally substituted by mono-, di-, or tri- halogen]; and
X represents O or CHz.
More preferably, said urea derivative of the formula (I) is
N-{2-[ethyl(3-methylphenyl)amino]ethyl}-N'-(3-oxo-3,4-dihydro-2H-1,4-
benzoxazin-6-yl)urea; or
N-{2-[ethyl(3-methylphenyl)amino]ethyl}-N'-(2-oxo-1,2,3,4-tetrahydroquinolin-5-
yl)urea.
The urea derivatives of formula (I), their tautomeric and stereoisomeric form,
and salts thereof
surprisingly show excellent VRl antagonistic activity. They are, therefore
suitable especially for
the prophylaxis and treatment of diseases associated with VRl activity, in
particular for the
treatment of urological diseases or disorders, such as detrusor overactivity
(overactive bladder),
urinary incontinence, neurogenic detrusor oeractivity (detrusor hyperflexia),
idiopathic detrusor
overactivity (detrusor instability), benign prostatic hyperplasia, and lower
urinary tract symptoms.
The compounds of the present invention are also effective for treating or
preventing a disease
selected from the group consisting of chronic pain, neuropathic pain,
postoperative pain,
rheumatoid arthritic pain, neuralgia, neuropathies, algesia, nerve injury,
ischaemia, neuro
degeneration and/or stroke, as well as inflammatory diseases such as asthma
and COPD since the
diseases also relate to VR1 activity.
The compounds of the present invention are also effective for treating or
preventing respiratory
diseases and inflammatory disorders such as common cold, cough, sneeze,
bronchitis including
acute and chronic bronchitis, bronchiolitis, rhinitis, allergic rhinitis,
vasomotor rhinitis, mucositis,
sinusitis, allergy, disorders associated with exogenous irritants such as
tobacco smoke, smog, high
levels of atmospheric SOZ and noxious gases in the workplace, and airways
hyperreactivity, milk
product intolerance, Loffler's pneumonia, emphysema, cystic fibrosis,
bronchiectasis, pulmonary
fibrosis, pneumoconiosis, collagen vascular disease, granulomatous disease,
laryngitis, pharyngitis,
pneumonia, pleuritis, persistent asthma and chronic asthmatic bronchitis since
the diseases also
relate to VRl activity.
The compounds of the present invention are also useful for the treatment and
prophylaxis of
neuropathic pain, which is a form of pain often associated with herpes zoster
and post-herpetic
neuralgia, painful diabetic neuropathy, neuropathic low back pain,
posttraumatic and postoperative
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neuralgia, neuralgia due to nerve compression and other neuralgias, phantom
pain, complex
regional pain syndromes, infectious or parainfectious neuropathies like those
associated with HIV
infection, pain associated with central nervous system disorders like multiple
sclerosis or
Parkinson disease or spinal cord injury or traumatic brain injury, and post-
stroke pain.
Furthermore, the compounds of the present invention are useful for the
treatment of
musculoskeletal pain, forms of pain often associated with osteoarthritis or
rheumatoid arthritis or
other forms of arthritis, and back pain.
In addition, the compounds of the present invention are useful for the
treatment of pain associated
with cancer, including visceral or neuropathic pain associated with cancer or
cancer treatment.
The compounds of the present invention are furthermore useful for the
treatment of visceral pain,
e.g. pain associated with obstruction of hollow viscus like gallstone colik,
pain associated with
irritable bowel syndrome, pelvic pain, vulvodynia, orchialgia or
prostatodynia, pain associated
with inflammatory lesions of joints, skin, muscles or nerves, and orofascial
pain and headache, e.g.
migraine or tension-type headache.
Further, the present invention provides a medicament, which includes one of
the compounds,
described above and optionally pharmaceutically acceptable excipients.
Alkyl per se and "alk" and "alkyl" in alkenyl, alkynyl, alkoxy, alkanoyl,
alkylamino, alkylamino-
carbonyl, alkylaminosulfonyl, alkylsulfonylamino, alkoxycarbonyl,
alkoxycarbonylamino and
alkanoylamino represent a linear or branched alkyl radical having generally 1
to 6, preferably 1 to
4 and particularly preferably 1 to 3 carbon atoms, representing illustratively
and preferably methyl,
ethyl, n-propyl, isopropyl, tent-butyl, n-pentyl and n-hexyl.
Alkoxy illustratively and preferably represents methoxy, ethoxy, n-propoxy,
isopropoxy, tert-
butoxy, n-pentoxy and n-hexoxy.
Alkylamino illustratively and preferably represents an alkylamino radical
having one or two
(independently selected) alkyl substituents, illustratively and preferably
representing methylamino,
ethylamino, n-propylamino, isopropylamino, tert-butylamino, n-pentylamino, n-
hexyl-amino, N,N-
dimethylamino, N,N-diethylamino, N-ethyl-N-methylamino, N-methyl-N-n-
propylamino,
N-isopropyl-N-n-propylamino, N-t-butyl-N-methylamino, N-ethyl-N-n-pentylamino
and N-n-
hexyl-N-methylamino. .
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Aryl per se and in arylamino and in arylcarbonyl represents a mono- to
tricyclic aromatic
carbocyclic radical having generally 6 to 14 carbon atoms, illustratively and
preferably
representing phenyl, naphthyl and phenanthrenyl.
Cycloalkyl per se and in cycloalkylamino and in cycloalkylcarbonyl represents
a cycloalkyl group
having generally 3 to 8 and preferably 5 to 7 carbon atoms, illustratively and
preferably
representing cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
Heteroaryl per se and the heteroaryl portion of the heteroaralkyl,
heteroaryloxy, heteroarallcyloxy,
or heteroarylcarbamoyl represent an aromatic mono- or bicyclic radical having
generally 5 to 10
and preferably 5 or 6 ring atoms and up to 5 and preferably up to 4 hetero
atoms selected from the
group consisting of S, O and N, illustratively and preferably representing
thienyl, furyl, pyrrolyl,
thiazolyl, oxazolyl, imidazolyl, pyridyl, pyrimidyl, pyridazinyl, indolyl,
isoindolino, indazolyl,
benzofuranyl, benzothiophenyl, quinolinyl, isoquinolinyl, tetrazolyl, and
triazolyl.
Heterocyclyl per se and in heterocyclylcarbonyl represents a mono- or
polycyclic, preferably
mono- or bicyclic, nonaromatic heterocyclic radical having generally 4 to 10
and preferably 5 to 8
ring atoms and up to 3 and preferably up to 2 hetero atoms and/or hetero
groups selected from the
group consisting of N, O, and S. The heterocyclyl radicals can be saturated or
partially
unsaturated. Preference is given to 5- to 8-membered monocyclic saturated
heterocyclyl radicals
having up to two hetero atoms selected from the group consisting of O, N and
S, such as
illustratively and preferably 1,3-dioxalanyl, tetrahydrofuran-2-yl, pyrrolidin-
2-yl, pyrrolidin-3-yl,
pyrrolinyl, piperidinyl, morpholinyl, perhydroazepinyl.
Ring per se represents a mono- or polycyclic, preferably mono- or bicyclic,
aromatic or
nonaromatic cyclic radical having generally 4 to 10 and preferably 5 to 8 ring
atoms and optionally
up to 3 and preferably up to 2 hetero atoms and/or hetero groups selected from
the group
consisting of N, O, and S. The cyclyl radicals can be saturated or partially
unsaturated. Preference
is given to 5- to 8-membered monocyclic saturated cyclyl radicals optionally
having up to two
hetero atoms selected from the group consisting of O, N and S. Ring
illustratively and preferably
includes aryl such phenyl, naphthyl and phenanthrenyl; cycloallcyl such as
cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl; heteroaryl such as
thienyI, furyl, pyrrolyl,
thiazolyl, oxazolyl, imidazolyl, pyridyl, pyrimidyl, pyridazinyl, indolyl,
isoindolino, indazolyl,
benzofuranyl, benzothiophenyl, quinolinyl, isoquinolinyl, tetrazolyl, and
triazolyl; and heterocyclic
such as 1,3-dioxalanyl, tetrahydrofuran-2-yl, pyrrolidin-2-yl, pyrrolidin-3-
yl, pyrrolinyl,
piperidinyl, morpholinyl, and perhydroazepinyl.
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EMBODIMENT OF THE INVENTION
The compound of the formula (I) of the present invention can be, but not
limited to be, prepared by
combining various known methods. In some embodiments, one or more of the
substituents, such
as amino group, carboxyl group, and hydroxyl group of the compounds used as
starting materials
or intermediates are advantageously protected by a protecting group known to
those skilled in the
art. Examples of the protecting groups are described in "Protective Groups in
Organic Synthesis
(3rd Edition)" by Greene and Wuts, John Wiley and Sons, New York 1999.
The compound of the formula (I) of the present invention can be, but not,
limited to be, prepared by
the Method [A], [B], [C], [D] or [E] below.
[Method A]
~ R~
H~N~N~R2 O
X NHz p / I (IV) X ~ ~ R~
N N~N~
/ L~ O ~ / H H ~RZ
O
H O H
(I)
(II) (III)
The compound of the formula (I) (wherein n, R', RZ and X are the same as
defined above) can be
prepared by reacting the compound of the formula (II) (wherein X is the same
as defined above)
and the compound of the formula (III) (wherein L~ represents a leaving group
including halogen
atom such as chlorine, bromine, or iodine atom) and then adding the compound
of the formula (IV)
(wherein n, R' and RZ are the same as defined above) to the reaction mixture.
The reaction may be carried out in a solvent including, for instance,
halogenated hydrocarbons
such as dichloromethane, chloroform and 1,2-dichloroethane; ethers such as
diethyl ether,
isopropyl ether, dioxane and tetrahydrofuran (THF) and 1,2-dimethoxyethane;
aromatic hydro-
carbons such as benzene, toluene and xylene; nitriles such as acetonitrile;
amides such as N, N-
dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-methylpyrrolidone
(NMP);
urea such as 1,3-dimethyl-2-imidazolidinone (DMI); sulfoxides such as
dimethylsulfoxide
(DMSO); and others. Optionally, two or more of the solvents selected from the
listed above can be
mixed and used.
The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about 20°C to
50°C. The reaction may be
conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10 hours.
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The reaction can be advantageously carried out in the presence of a base
including, for instance,
organic amines such as pyridine, triethylamine and N,N-diisopropylethylamine,
dimethylaniline,
diethylaniline, 4-dimethylaminopyridine, and others.
The compound of the formula (III) and (IV) are commercially available or can
be prepared by the
use of known techniques.
[Method B]
O
X NH2 ~ X
,R \ ~ ~R
/ + OC n N~RZ -~ I N N~N~ Z
/~ ~ H H R
O N
H
l) M U)
The compound of the formula (I) can be prepared by the reaction of the
compound of the formula
(II) and the compound of the formula (V) (wherein n, R' and RZ are the same as
defined above).
The reaction may be carried out in a solvent including, for instance,
halogenated hydrocarbons
such as dichloromethane, chloroform and 1,2-dichloroethane; ethers such as
diethyl ether,
isopropyl ether, dioxane and tetrahydrofuran (THF) and 1,2-dimethoxyethane;
aromatic
hydrocarbons such as benzene, toluene and xylene; nitriles such as
acetonitrile; amides such as N,
N-dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-
methylpyrrolidone
(NMP); urea such as 1,3-dimethyl-2-imidazolidinone (DMI); sulfoxides such as
dimethylsulfoxide
(DMSO); and others. Optionally, two or more of the solvents selected from the
listed above can be
mixed and used.
The reaction can be carried out in the presence of organic base such as
pyridine or triethylamine.
The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about room temperature to
100°C. The reaction
may be conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10
hours.
The compound (V) can be prepared by the use of known techniques or are
commercially available.
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[Method C]
j~ R'
H~N~N~ ~ O
phosgene, R
X NHZ diphosgene, ~lV) X \ ~ ~R~
triphosgene, ~ ~ ~ / H H~N~R2
O H CDI or CDT O H
The compound of the formula (I) can be prepared by reacting the compound of
the formula (II)
with phosgene, diphosgene, triphosgene, 1,1-carbonyldiimidazole (CDI), or 1,1'-
carbonyldi(1,2,4-
triazole)(CDT), and then adding the compound of the formula (1V) to the
reaction mixture.
The reaction may be carried out in a solvent including, for instance,
halogenated hydrocarbons
such as dichloromethane, chloroform and 1,2-dichloroethane; ethers such as
diethyl ether,
isopropyl ether, dioxane and tetrahydrofuran (THF) and 1,2-dimethoxyethane;
aromatic
hydrocarbons such as benzene, toluene and xylene; nitriles such as
acetonitrile; amides such as N,
N-dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-
methylpyrrolidone
(NMP); urea such as 1,3-dimethyl-2-imidazolidinone (DMI); sulfoxides such as
dimethylsulfoxide
(DMSO); and others. Optionally, two or more of the solvents selected from the
listed above can be
mixed and used.
The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about 20°C to
50°C. The reaction may be
conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10 hours.
Phosgene, diphosgene, triphosgene, CDI, and CDT are commercially available.
[Method D]
X NHZ
O N O
phosgene, H ~ ~ R'
H N n NCR diphosgene, III) X N H~N~R2
~Rz + triphosgene,
CDI or CDT O N
H
CI)
The compound of the formula (I) can be prepared by reacting the compound of
the formula (IV)
with phosgene, diphosgene, triphosgene, 1,1-carbonyldiimidazole (CDIJ, or 1,1'-
carbonyldi(1,2,4-
triazole)(CDT) and then adding the compound of the formula (II) to the
reaction mixture.
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The reaction may be carried out in a solvent including, for instance,
halogenated hydrocarbons
such as dichloromethane, chloroform and 1,2-dichloroethane; ethers such as
diethyl ether,
isopropyl ether, dioxane and tetrahydrofuran (THF) and 1,2-dimethoxyethane;
aromatic hydro-
carbons such as benzene, toluene and xylene; nitriles such as acetonitrile;
amides such as N, N-
dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-methylpyrrolidone
(NMP);
urea such as 1,3-dimethyl-2-imidazolidinone (DMI); sulfoxides such as
dimethylsulfoxide
(DMSO); and others. Optionally, two or more of the solvents selected from the
listed above can be
mixed and used.
The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about 20°C to
50°C. The reaction may be
conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10 hours.
[Method E]
X NHZ
O N
H O
~R~ + O ~ ~ (II) X ~ ~ R~
~ N N~N~
HZN~N~RZ L~ O ~ ( / H H ~RZ
O N
(I~ (III) H (I)
The compound of the formula (I) can be prepared by reacting the compound of
the formula (IV)
and the compound of the formula (III), and then adding 'the compound of the
formula (II) to the
reaction mixture.
The reaction may be carried out in a solvent including, for instance,
halogenated hydrocarbons
such as dichloromethane, chloroform and 1,2-dichloroethane; ethers such as
diethyl ether,
isopropyl ether, dioxane and tetrahydrofuran (THF) and 1,2-dimethoxyethane;
aromatic
hydrocarbons such as benzene, toluene and xylene; nitrites such as
acetonitrile; amides such as N,
N-dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-
methylpyrrolidone
(NMP); urea such as 1,3-dimethyl-2-imidazolidinone (DMI); sulfoxides such as
dimethylsulfoxide
(DMSO); and others. Optionally, two or more of the solvents selected from the
listed above can be
mixed and used.
The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about 20°C to
SO°C. The reaction may be
conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10 hours.
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The reaction can be advantageously carried out in the presence of a base
including, for instance,
organic amines such as pyridine, triethylamine and N,N-diisopropylethylamine,
dimethylaniline,
diethylaniline, 4-dimethylaminopyridine, and others.
Preparation of compound of the formula (II)
The compounds of the formula (II) are commercially available or can be
prepared by well known
methods. For example, the compounds of the formula (II') can be prepared by
the following
procedures.
O
NOz ~CI H H
CI
I \ (VIII) O N \ O N \
---~ I NOz --.~ I NHz
HzN ~ Step 1 X' / Step 2
Y
(VI) NII) (II')
In the Step l, the compound of the formula (VII) (wherein X' represent O, S,
or NRl°) can be
prepared by cyclization of the compound of the formula (VI) (wherein Y
represents OH, SH, or
NH(RI°) ) using the agent including, for instance, chloroacetyl
chloride.
The reaction can be carried out in a solvent including, for instance, amides
such as N, N-
dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-methylpyrrolidone
(NMP),
and the like. Optionally, two or more of the solvents selected from the listed
above can be mixed
and used.
The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about 0°C to
80°C. The reaction may be
conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10 hours.
In the Step 2, the compound of the formula (II') can be prepared by the by the
reduction of the
compound of the formula (VI>] by hydrogenation using a catalyst including, for
instance,
palladium on carbon and platinum on carbon.
The reaction can be carried out in a solvent including, for instance, amides
such as N, N-
dimethylformamide (DMF), N, N-dimethylacetamide (DMAC) and N-methylpyrrolidone
(NMP)
and others.
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The reaction temperature can be optionally set depending on the compounds to
be reacted. The
reaction temperature is usually, but not limited to, about 20°C to
120°C. The reaction may be
conducted for, usually, 30 minutes to 24 hours and preferably 1 to 10 hours.
The compound (Vn is commercially available or can be prepared by the use of
known techniques.
The compound (II") below can be prepared, for example, with the use of wittig
reaction and
hydrogenation starting from dinitrobenzaldehyde
O N CHO-COOH O
z
NOz I / NHz
OHC
(VIII)
(I I,.)
When the compound shown by the formula (I) or a salt thereof has an asymmetric
carbon in the
structure, their optically active compounds and racemic mixtures are also
included in the scope of
the present invention.
Typical salts of the compound shown by the formula (I) include salts prepared
by reaction of the
compounds of the present invention with a mineral or organic acid, or an
organic or inorganic
base. Such salts are known as acid addition and base addition salts,
respectively.
Acids to form acid addition salts include inorganic acids such as, without
limitation, sulfuric acid,
phosphoric acid, hydrochloric acid, hydrobromic acid, hydriodic acid and the
like, and organic
acids, such as, without limitation, p-toluenesulfonic acid, methanesulfonic
acid, oxalic acid, p-
bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic
acid, acetic acid, and
the like.
Base addition salts include those derived from inorganic bases, such as,
without limitation,
ammonium hydroxide, alkaline metal hydroxide, alkaline earth metal hydroxides,
carbonates,
bicarbonates, and the like, and organic bases, such as, without limitation,
ethanolamine,
triethylamine, tris(hydroxymethyl)aminomethane, and the like. Examples of
inorganic bases
include sodium hydroxide, potassium hydroxide, potassium carbonate, sodium
carbonate, sodium
bicarbonate, potassium bicarbonate, calcium hydroxide, calcium carbonate, and
the like.
The compound of the present invention or a salt thereof, depending on its
substituents, may be
modified to form lower alkylesters or known other esters; and/or hydrates or
other solvates. Those
esters, hydrates, and solvates are included in the scope of the present
invention.
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The compound of the present invention may be administered in oral forms, such
as, without
limitation, normal and enteric coated tablets, capsules, pills, powders,
granules, elixirs, tinctures,
solution, suspensions, syrups, solid and liquid aerosols and emulsions. They
may also be
administered in parenteral forms, such as, without limitation, intravenous,
intraperitoneal,
subcutaneous, intramuscular, and the like forms, well-known to those of
ordinary skill in the
pharmaceutical arts. The compounds of the present invention can be
administered in intranasal
form via topical use of suitable intranasal vehicles, or via transdermal
routes, using transdermal
delivery systems well-known to those of ordinary skilled in the art, or
perlingual or buccal or via
inhalation.
The dosage regimen with the use of the compounds of the present invention is
selected by one of
ordinary skill in the arts, in view of a variety of factors, including,
without limitation, age, weight,
sex, and medical condition of the recipient, the severity of the condition to
be treated, the route of
administration, the level of metabolic and excretory function of the
recipient, the dosage form
employed, the particular compound and salt thereof employed.
The compounds of the present invention are preferably formulated prior to
administration together
with one or more pharmaceutically-acceptable excipients. Excipients are inert
substances such as,
without limitation carriers, diluents, flavoring agents, sweeteners,
lubricants, solubilizers, suspend-
ding agents, binders, tablet disintegrating agents and encapsulating material.
Yet another embodiment of the present invention is pharmaceutical formulation
comprising a
compound of the invention and one or more pharmaceutically-acceptable
excipients that are
compatible with the other ingredients of the formulation and not deleterious
to the recipient
thereof. Pharmaceutical formulations of the invention are prepared by
combining a therapeutically
effective amount of the compounds of the invention together with one or more
pharmaceutically-
acceptable excipients therefore. In making the compositions of the present
invention, the active
ingredient may be mixed with a diluent, or enclosed within a carrier, which
may be in the form of a
capsule, sachet, paper, or other container. The carrier may serve as a
diluent, which may be solid,
semi-solid, or liquid material which acts as a vehicle, or can be in the form
of tablets, pills
powders, lozenges, elixirs, suspensions, emulsions, solutions, syrups,
aerosols, ointments,
containing, for example, up to 10% by weight of the active compound, soft and
hard gelatin
capsules, suppositories, sterile injectable solutions and sterile packaged
powders.
For oral administration, the active ingredient may be combined with an oral,
and non-toxic,
pharmaceutically-acceptable carrier, such as, without limitation, lactose,
starch, sucrose, glucose,
sodium carbonate, mannitol, sorbitol, calcium carbonate, calcium phosphate,
calcium sulfate,
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methyl cellulose, and the like; together with, optionally, disintegrating
agents, such as, without
limitation, maize, starch, methyl cellulose, agar bentonite, xanthan gum,
alginic acid, and the like;
and optionally, binding agents, for example, without limitation, gelatin,
natural sugars, beta-
lactose, corn sweeteners, natural and synthetic gums, acacia, tragacanth,
sodium alginate,
carboxymethylcellulose, polyethylene glycol, waxes, and the like; and,
optionally, lubricating
agents, for example, without limitation, magnesium stearate, sodium stearate,
stearic acid, sodium
oleate, sodium benzoate, sodium acetate, sodium chloride, talc, and the like.
In powder forms, the carrier may be a finely divided solid which is in
admixture with the finely
divided active ingredient. The active ingredient may be mixed with a carrier
having binding
properties in suitable proportions and compacted in the shape and size desired
to produce tablets.
The powders and tablets preferably contain from about 1 to about 99 weight
percent of the active
ingredient which is the novel composition of the present invention. Suitable
solid carriers are
magnesium carboxymethyl cellulose, low melting waxes, and cocoa butter.
Sterile liquid formulations include suspensions, emulsions, syrups and
elixirs. The active
ingredient can be dissolved or suspended in a pharmaceutically acceptable
carriers, such as sterile
water, sterile organic solvent, or a mixture of both sterile water and sterile
organic solvent.
The active ingredient can also be dissolved in a suitable organic solvent, for
example, aqueous
propylene glycol. Other compositions can be made by dispersing the finely
divided active
ingredient in aqueous starch or sodium carboxymethyl cellulose solution or in
a suitable oil.
For intranasal administration, the pharmaceutical compositions of this
invention may be
administered by nasal drops, by nasal aerosols, or as an inhaled powder.
Suitable nasal spray formulations of inventive compositions can be readily
prepared according to
techniques well known in the art of pharmaceutical formulation. For example,
the preparation of
solutions or emulsions are described by Achari et al., U.S. Pat. No. 6,
436,950 (supra), J. G. Nair
[Chapt. 39, Solutions, Emulsions, Suspensions and Extracts, pg. 721-752J] and
aerosols by J.
Sciarra and C. J. Sicarra [Chapt. 50, "Aerosols", pg. 963 to 979] in the
standard text: "Remington,
the science and practice of pharmacy," Alfonso R. Gennaro, Chairman of the
editorial board and
editor. 20th ed. Baltimore, Md. Lippincott Williams & Wilkins, 2000.
The compositions the active ingredient may be ,prepared as gels, liposomal
dispersions,
suspensions or emulsions in saline, employing benzyl alcohol, benzalkonium
chloride or other
suitable preservatives, absorption promoters such as cyclodextrins to enhance
bioavailability and
bioadhesives for prolonged contact, and/or other solubilizing or dispersing
agents known in the art.
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Thus, a composition for administration to the intranasal surfaces is
particularly contemplated that
comprises a solution of the active ingredient dissolved or dispersed in a
pharmaceutically
acceptable diluent (carrier). The solvent or wetting agent may be propylene
glycol (1,2-
propanediol) and a variety of aqueous carriers can be used, e.g. buffered
water, 0.9 percent saline,
buffered aqueous-ethanol solutions and the like. These compositions can be
sterilized by
conveiltional, well-known sterilization techniques, or can be sterile
filtered. The resulting solutions
can be packaged for use as is or mixed as an adjuvant to another medication.
The inventive embodiment compositions can contain pharmaceutically acceptable
auxiliary
substances as required to approximate physiological conditions, such as pH
adjusting and
buffering agents, tonicity adjusting agents, wetting agents and the like, for
example, sodium
acetate, sodium lactate, sodium chloride, potassium chloride, calcium
chloride, sorbitan
monolaurate, triethanolamine oleate, and the like.
Another form of intranasal administration is to administer the active
ingredient in powder form; by
itself or admixed to an inert carrier such as calcium carbonate or lactose.
Methods for preparing
spray dried powder with a hydrophilic excipient, e.g. povidone, lactose, and
delivering it using dry
powder nasal inhalers, have been described by Gordon et al. (U.S. Pat. No.
6,365,190) and are
incorporated herein by reference. The advantage of a powder method for
delivery is that it may
have a more prolonged action when administered in dry powder versus in soluble
forms, as the
nose has robust clearance mechanisms. The powder may be prepared in micronized
form, by re-
crystallization, by granulation, by drying, or by milling to a specified
particle size and thus to have
a high surface area for interaction with cold receptors. Methods for preparing
powders are well-
known to the art and have been reviewed by R. E. O'Connor and J. B. Schwartz
(Powders, Chapt.
37, pg. 681-699) in the standard text: "Remington, the science and practice of
pharmacy," Alfonso
R. Gennaro, Chairman of the editorial board and editor. 20th ed. Baltimore,
Md. Lippincott
Williams & Wilkins, 2000. To quote from this Chapter (pg. 688):
The formulation may be in unit dosage form, which is a physically discrete
unit containing a unit
dose, suitable for administration in human or other mammals. A unit dosage
form can be a capsule
or tablets, or a number of capsules or tablets. A "unit dose" is a
predetermined quantity of the
active compound of the present invention, calculated to produce the desired
therapeutic effect, in
association with one or more excipients. The quantity of active ingredient in
a unit dose may be
varied or adjusted from about 0.1 to about 1000 milligrams or more according
to the particular
treatment involved.
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Typical oral dosages of the present invention, when used for the indicated
effects, will range from
about O.Olmg /kg/day to about 100 mg/lcg/day, preferably from 0.1 mg/kg/day to
30 mg/kg/day,
and most preferably from about 0.5 mg/kg/day to about 10 mg/kg/day. In the
case of parenteral
administration, it has generally proven advantageous to administer quantities
of about 0.001 to
100mg /kg/day, preferably from 0.01 mg/kg/day to 1 mg/kg/day. The compounds of
the present
invention may be administered in a single daily dose, or the total daily dose
may be administered
in divided doses, two, three, or more times per day. Where delivery is via
transdermal forms, of
course, administration is continuous.
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EXAMPLES
The present invention will be described as a form of examples, but they should
by no means be
construed as defining the metes and bounds of the present invention.
In the examples below, all quantitative data, if not stated otherwise, relate
to percentages by
weight.
Mass spectra were obtained using electrospray (ES) ionization techniques
(micromass Platform
LC). Melting points are uncorrected. Liquid Chromatography - Mass spectroscopy
(LC-MS) data
were recorded on a Micromass Platform LC with Shimadzu Phenomenex ODS
column(4.6 mm~ X
30 mm) flushing a mixture of acetonitrile-water (9:1 to 1:9) at 1 ml/min of
the flow rate. TLC was
performed on a precoated silica gel plate (Merck silica gel 60 F-254). Silica
gel (WAKO-gel
C-200 (75-150 Vim)) was used for all column chromatography separations. All
chemicals were
reagent grade and were purchased from Sigma-Aldrich, Wako pure chemical
industries, Ltd., Great
Britain, Tokyo kasei kogyo Co., Ltd., Nacalai tesque, Inc., Watanabe Chemical
Ind. Ltd.,
Maybridge plc, Lancaster Synthesis Ltd., Merck KgaA, Germany, Kanto Chemical
Co., Ltd.
'H NMR spectra were recorded using either Bruker DRX-300 (300 MHz for'H)
spectrometer or
Brucker 500 UltraShieled~ (500 MHz for 1H) . Chemical shifts are reported in
parts per million
(ppm) with tetramethylsilane (TMS) as an internal standard at zero ppm.
Coupling constant (J) are
given in hertz and the abbreviations s, d, t, q, m, and br refer to singlet,
doblet, triplet, quartet,
multiplet, and broad, respectively. The mass determinations were carried out
by MAT95 (Finnigan
MAT).
All starting materials are commercially available or can be prepared using
methods cited in the
literature.
The effect of the present compounds was examined by the following assays and
pharmacological
tests.
[Measurement of capsaicin-induced Caa+ influx in the human VRl-transfected CHO
cell line]
(Assay 1)
(1) Establishment ofthe human VRl-CHOluc9aeq cell line
Human vanilloid receptor (hVRI) cDNA was cloned from libraries of axotomized
dorsal
root ganglia (WO 00/29577). The cloned hVRI cDNA was constructed with pcDNA3
vector and transfected into a CHOluc9aeq cell line. The cell line contains
aequorin and
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CRE-luciferase reporter genes as read-out signals. The transfectants were
cloned by
limiting dilution in selection medium (DMEM/F12 medium (Gibco BRL)
supplemented
with 10% FCS, 1.4 mM Sodium pyruvate, 20 mM HEPES, 0.15% Sodium bicarbonate,
100 U/ml penicillin, 100 p,g/ml streptomycin, 2 mM glutamine, non-essential
amino acids
and 2 mg/ml G418). Caz+ influx was examined in the capsaicin-stimulated
clones. A high
responder clone was selected and used for further experiments in the project.
The human
VRl-CHOluc9aeq cells were maintained in the selection medium and passaged
every 3-4
days at 1-2.5x105 cells/flask (75 mmz).
(2) Measurement of Ca2+ influx using FDSS-3000
Human VRl-CHOluc9aeq cells were suspended in a culture medium which is the
same as
the selection medium except for 6418 and seeded at a density of 1,000 cells
per well into
384-well plates (black walled clear-base / Nalge Nunc International).
Following the culture
for 48 hrs the medium was changed to 2 p,M Fluo-3 AM (Molecular Probes) and
0.02%
Puronic F-127 in assay buffer (Hank's balanced salt solution (HBSS), 17 mM
HEPES
(pH7.4), 1 mM Probenecid, 0.1% BSA) and the cells were incubated for 60 min at
25°C.
After washing twice with assay buffer the cells were incubated with a test
compound or
vehicle for 20 min at 25°C. Mobilization of cytoplasmic Caz+ was
measured by FDSS-3000
(~.eX 488nm, 7~em 540nm / Hamamatsu Photonics) for 60 sec after the
stimulation with 10
nM capsaicin. Integral R was calculated and compared with controls.
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[Measurement of the capsaicin-induced Caz+ influx in primary cultured rat
dorsal root ganglia
neurons] (Assay 2)
(1) Preparation of rat dorsal root ganglia neurons
New born Wister rats (5-11 days) were sacrificed and dorsal root ganglia (DRG)
was
removed. DRG was incubated with 0.1% trypsin (Gibco BRL) in PBS(-) (Gibco BRL)
for
30 min at 37°C, then a half volume of fetal calf serum (FCS) was added
and the cells were
spun down. The DRG neuron cells were resuspended in Ham F12/5% FCS/5% horse
serum (Gibco BRL) and dispersed by repeated pipetting and passing through 70
p.m mesh
(Falcon). The culture plate was incubated for 3 hours at 37°C to remove
contaminating
Schwann cells. Non-adherent cells were recovered and further cultured in
laminin-coated
384 well plates (Nunc) at 1x10 cells/50 ~1/well for 2 days in the presence of
50 ng/ml
recombinant rat NGF (Sigma) and 50 p,M 5-fluorodeoxyuridine (Sigma).
(2) Ca2+ mobilization assay
DRG neuron cells were washed twice with HBSS supplemented with 17 mM HEPES (pH
7.4) and 0.1% BSA. After incubating with 2 pM fluo-3AM (Molecular Probe),
0.02%
PF127 (Gibco BRL) and 1 mM probenecid (Sigma) for 40 min at 37°C, cells
were washed
3 times. The cells were incubated with VR1 antagonists or vehicle
(dimethylsulfoxide) and
then with 1 pM capsaicin in FDSS-6000 (~,eX 480nm, ~,em 520nm / Hamamatsu
Photonics). The fluorescence changes at 480nm were monitored for 2.5 min.
Integral R
was calculated and compared with controls.
[Organ bath assay to measure the capsaicin-induced bladder contraction] (Assay
3)
Male Wistar rats (10 week old) were anesthetized with ether and sacrificed by
dislocating
the necks. The whole urinary bladder was excised and placed in oxygenated
Modified
Krebs-Henseleit solution (pH 7.4) of the following composition (112mM NaCI,
5.9mM
KCI, l.2mM MgClz, l.2mM NaHZP04, 2mM CaCl2, 2.SmM NaHC03, l2mM glucose).
Contractile responses of the urinary bladder were studied as described
previously [Maggi
CA et al: Br.J.Pharmacol. 108: 801-805, 1993]. Isometric tension was recorded
under a
load of 1 g using longitudinal strips of rat detrusor muscle. Bladder strips
were
equilibrated for 60 min before each stimulation. Contractile response to 80 mM
KCl was
determined at 15 min intervals until reproducible responses were obtained. The
response to
KCl was used as an internal standard to evaluate the maximal response to
capsaicin. The
effects of the compounds were investigated by incubating the strips with
compounds for 30
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_29_
min prior to the stimulation with 1 ~M capsaicin (vehicle: 80% saline, 10%
EtOH, and
10% Tween 80). One of the preparations made from the same animal was served as
a
control while the others were used for evaluating compounds. Ratio of each
capsaicin-
induced contraction to the internal standard (i.e. KCl-induced contraction)
was calculated
and the effects of the test compounds on the capsaicin-induced contraction
were evaluated.
[Measurement of Caz+ influx in the human P2X1-transfected CHO cell line]
(1) Preparation of the human P2X1-transfected CHOluc9aeq cell line
Human P2X1-transfected CHOluc9aeq cell line was established and maintained in
Dulbecco's modified Eagle's medium (DMEM/F12) supplemented with 7.5% FCS, 20
mM HEPES-KOH (pH 7.4), 1.4 mM sodium pyruvate, 100 U/ml penicillin, 100 p.g/ml
streptomycin, 2 mM glutamine (Gibco BRL) and 0.5 Units/ml apyrase (grade I,
Sigma).
The suspended cells were seeded in each well of 384-well optical bottom black
plates
(Nalge Nunc International) at 3 x 103 / 50 ~1 / well. The cells were cultured
for following
48 hrs to adhere to the plates.
(2) Measurement of the intracellular Ca2+ levels
P2X1 receptor agonist-mediated increases in cytosolic Ca2+ levels were
measured using a
fluorescent Ca2+ chelating dye, Fluo-3 AM (Molecular Probes). The plate-
attached cells
were washed twice with washing buffer (HBSS, 17 mM HEPES-KOH (pH 7.4), 0.1%
BSA
and 0.5 units/ml apyrase), and incubated in 40 ~,1 of loading buffer (1 pM
Fluo-3 AM, 1
mM probenecid, 1 ~M cyclosporin A, 0.01% pluronic (Molecular Probes)in washing
buffer) for 1 hour in a dark place. The plates were washed twice with 40 pl
washing buffer
and 35 ~,1 of washing buffer were added in each well with 5 ~1 of test
compounds or 2',3'-
0-(2,4,6-trinitrophenyl) adenosine 5'-triphpsphate (Molecular Probes) as a
reference. After
further incubation for 10 minutes in dark 200 nM a, (3-methylene ATP agonist
was added
to initiate the Ca2+ mobilization. Fluorescence intensity was measured by FDSS-
6000
(~eX 410nm, ~,e"~ S l Onm / Hamamatsu Photonics) at 250 msec intervals.
Integral ratios
were calculated from the data and compared with that of a control.
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[Measurement of capsaicin-induced bladder contraction in anesthetized rats]
(Assay 4)
(1) Animals
Female Sprague-Dawley rats (200250 g / Charles River Japan) were used.
(2) Catheter implantation
Rats were anesthetized by intraperitoneal administration of urethane (Sigma)
at 1.2 g/kg.
The abdomen was opened through a midline incision, and a polyethylene catheter
(BECTON DICKINSON, PE50) was implanted into the bladder through the dome. In
parallel, the inguinal region was incised, and a polyethylene catheter
(Hibiki, size 5) filled
with 2 ICT / ml of heparin (Novo Heparin, Aventis Pharma) in saline (Otsuka)
was inserted
into a common iliac artery.
(3) Cystometric investigation
The bladder catheter was connected via T-tube to a pressure transducer (Viggo-
Spectramed Pte Ltd, DT-XXAD) and a microinjection pump (TERUMO). Saline was
infused at room temperature into the bladder at a rate of 2.4 ml/hr.
Intravesical pressure
was recorded continuously on a chart pen recorder (Yokogawa). At least three
reproducible micturition cycles, corresponding to a 20-minute period, were
recorded
before a test compound administration and used as baseline values.
(4) Administration of test compounds and stimulation of bladder with capsaicin
The saline infusion was stopped before administrating compounds. A testing
compound
dissolved in the mixture of ethanol, Tween 80 (ICN Biomedicals Inc.) and
saline (1 : 1 : 8,
v/v/v) was administered intraarterially at 10 mg/kg. 2min after the
administration of the
compound 10 ~g of capsaicin (Nacalai Tesque) dissolved in ethanol was
administered
intraarterially.
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(5) Analysis of cystometry parameters
Relative increases in the capsaicin-induced intravesical pressure were
analyzed from the
cystometry data. The capsaicin-induced bladder pressures were compared with
the
maximum bladder pressure during micturition without the capsaicin stimulation.
The
testing compounds-mediated inhibition of the increased bladder pressures was
evaluated
using Student's t-test. A probability level less than 5% was accepted as
significant
difference.
[Measurement of over active bladder in anesthetized cystitis rats] (Assay 5)
(1) Animals
~10 Female Sprague-Dawley rats (180250 g l Charles River Japan) were used.
Cyclo-
phosphamide (CYP) dissolved in saline was administered intraperitoneally at
150 mg/kg
48 hours before experiment.
(2) Catheter implantation
Rats were anesthetized by intraperitoneal administration of urethane (Sigma)
at 1.25 g/kg.
The abdomen was opened through a midline incision, and a polyethylene catheter
(BECTON DICKINSON, PE50) was implanted into the bladder through the dome. In
parallel, the inguinal region was incised, and a polyethylene catheter (BECTON
DICKINSON, PE50) filled with saline (Otsuka) was inserted into a femoral vein.
After the
bladder was emptied, the rats were left for 1 hour for recovery from the
operation.
(3) Cystometric investigation
The bladder catheter was connected via T-tube to a pressure transducer (Viggo-
Spectramed Pte Ltd, DT-XXAD) and a microinjection pump (TERUMO). Saline was
infused at room temperature into the bladder at a rate of 3.6 ml/hr for 20
min. Intravesical
pressure was recorded continuously on a chart pen recorder (Yokogawa). At
least three
reproducible micturition cycles, corresponding to a 20-minute period, were
recorded
before a test compound administration.
(4) Administration of test compounds
A testing compound dissolved in the mixture of ethanol, Tween 80 (ICN
Biomedicals Inc.)
and saline (1 : 1 : 8, v/vlv) was administered intravenously at 0.05 mg/kg,
0.5 mg/kg or S
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mg/lcg. 3min after the administration of the compound, saline (Nacalai Tesque)
was
infused at room temperature into the bladder at a rate of 3.6 ml/hr.
(5) Analysis of cystometry parameters
The cystometry parameters were analyzed as described previously [ Lecci A et
al: Eur. J.
Pharmacol. 259: 129-135, 1994]. The micturition frequency calculated from
micturition
interval and the bladder capacity calculated from a volume of infused saline
until the first
micturition were analyzed from the cystometry data. The testing compounds-
mediated
inhibition of the frequency and the testing compounds-mediated increase of
bladder
capacity were evaluated using unpaired Student's t-test. A probability levels
less than 5%
was accepted as significant difference. Data were analyzed as the mean + SEM
from 4 - 7
rats.
[Measurement of Acute Pain]
Acute pain is measured on a hot plate mainly in rats. Two variants of hot
plate testing are
used: In the classical variant animals are put on a hot surface (52 to
56°C) and the latency
time is measured until the animals show nociceptive behavior, such as stepping
or foot
licking. The other variant is an increasing temperature hot plate where the
experimental
animals are put on a surface of neutral temperature. Subsequently this surface
is slowly but
constantly heated until the animals begin to lick a hind paw. The temperature
which is
reached when hind paw licking begins is a measure for pain threshold.
Compounds are tested against a vehicle treated control group. Substance
application is
performed at different time points via different application routes (i.v.,
i.p., p.o., i.t., i.c.v.,
s.c., intradermal, transdermal) prior to pain testing.
[Measurement of Persistent Pain]
Persistent pain is measured with the formalin or capsaicin test, mainly in
rats. A solution
of 1 to 5% formalin or 10 to 100 ~g capsaicin is injected into one hind paw of
the
experimental animal. After formalin or capsaicin application the animals show
nociceptive
reactions like flinching, licking and biting of the affected paw. The number
of nociceptive
reactions within a time frame of up to 90 minutes is a measure for intensity
of pain.
Compounds are tested against a vehicle treated control group. Substance
application is
performed at different time points via different application routes (i.v.,
i.p., p.o., i.t., i.c.v.,
s.c., intradermal, transdermal) prior to formalin or capsaicin administration.
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[Measurement of Neuropathic Pain]
Neuropathic pain is induced by different variants of unilateral sciatic nerve
injury mainly
in rats. The operation is performed under anesthesia. The first variant of
sciatic nerve
injury is produced by placing loosely constrictive ligatures around the common
sciatic
nerve (Bennett and Xie, Pain 33 (1988): 87-107). The second variant is the
tight ligation of
about the half of the diameter of the common sciatic nerve (Seltzer et al.,
Pain 43 (1990):
205-218). In the next variant, a group of models is used in which tight
ligations or
transections are made of either the LS and L6 spinal nerves, or the LS spinal
nerve only
(KIM SH; CHUNG JM, AN EXPERIMENTAL-MODEL FOR PERIPHERAL
NEUROPATHY PRODUCED BY SEGMENTAL SPINAL NERVE LIGATION IN THE
RA, PAIN SO (3) (1992): 355-363). The fourth variant involves an axotomy of
two of the
three terminal branches of the sciatic nerve (tibial and common peroneal
nerves) leaving
the remaining sural nerve intact whereas the last variant comprises the
axotomy of only the
tibial branch leaving the sural and common nerves uninjured. Control animals
are treated
with a sham operation.
Postoperatively, the nerve injured animals develop a chronic mechanical
allodynia, cold
allodynia, as well as a thermal hyperalgesia. Mechanical allodynia is measured
by means
of a pressure transducer (electronic von Frey Anesthesiometer, IITC Inc.-Life
Science
Instruments, Woodland Hills, SA, USA; Electronic von Frey System, Somedic
Sales AB,
Horby, Sweden). Thermal hyperalgesia is measured by means of a radiant heat
source
(Plantar Test, Ugo Basile, Comerio, Italy), or by means of a cold plate of 5
to 10 °C where
the nocifensive reactions of the affected hind paw are counted as a measure of
pain
intensity. A further test for cold induced pain is the counting of nocifensive
reactions, or
duration of nocifensive responses after plantar administration of acetone to
the affected
hind limb. Chronic pain in general is assessed by registering the circadanian
rhytms in
activity (Surjo and Arndt, Universitat zu Koln, Cologne, Germany), and by
scoring
differences in gait (foot print patterns; FOOTPRINTS program, Klapdor et al.,
1997. A
low cost method to analyse footprint patterns. J. Neurosci. Methods 75, 49-
54).
Compounds are tested against sham operated and vehicle treated control groups.
Substance
application is performed at different time points via different application
routes (i.v., i.p.,
p.o., i.t., i.c.v., s.c., intradermal, transdermal) prior to pain testing.
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[Measurement of Inflammatory Pain]
Inflammatory pain is induced mainly in rats by injection of 0.75 mg
carrageenan or
complete Freund's adjuvant into one hind paw. The animals develop an edema
with
mechanical allodynia as well as thermal hyperalgesia. Mechanical allodynia is
measured
by means of a pressure transducer (electronic von Frey Anesthesiometer, IITC
Inc.-Life
Science Instruments, Woodland Hills, SA, USA). Thermal hyperalgesia is
measured by
means of a radiant heat source (Plantar Test, Ugo Basile, Comerio, Italy, Paw
thermal
stimulator, G. Ozaki, University of California, USA). For edema measurement
two
methods are being used. In the first method, the animals are sacrificed and
the affected
hindpaws sectioned and weighed. The second method comprises differences in paw
volume by measuring water displacement in a plethysmometer (Ugo Basile,
Comerio,
Italy).
Compounds are tested against uninflamed as well as vehicle treated control
groups.
Substance application is performed at different time points via different
application routes
(i.v., i.p., p.o., i.t., i.c.v., s.c., intradermal, transdermal) prior to pain
testing.
[Measurement of Diabetic Neuropathic Pain]
Rats treated with a single intraperitoneal injection of 50 to 80 mg/kg
streptozotocin
develop a profound hyperglycemia and mechanical allodynia within 1 to 3 weeks.
Mechanical allodynia is measured by means of a pressure transducer (electronic
von Frey
Anesthesiometer, IITC Inc.-Life Science Instruments, Woodland Hills, SA, USA).
Compounds are tested against diabetic and non-diabetic vehicle treated control
groups.
Substance application is performed at different time points via different
application routes
(i.v., i.p., p.o., i.t., i.c.v., s.c., intradermal, transdermal) prior to pain
testing.
Results in capsaicin-induced Caz+ influx assay in the human VRl-transfected
CHO cell line (Assay
1 ) are shown in Examples.
The compounds of the present invention also show excellent selectivity, and
strong activity in
other assays 2-S and assays for pain described above.
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Preparing method of compounds
[Example 1-1]
Reaction 1
N02
O O N ~ N02
H2N ~ + CI~CI
OH O
1
To a solution of 2-amino-4-nitrophenol (3.08g, 20.Ommol) and pyridine (l.S8g,
20.Ommol)
in DMF was added chloroacetyl chloride (2.26g, 20.Ommo1) at 0°C and the
mixture was stirred for
lh. NaH (60% in oil, 0.80g, 20.Ommo1) was added to the reaction mixture at
0°C and the mixture
was stirred for 2h. The reaction was quenched by addition of water and
extracted with ethyl
acetate (EtOAc). The combined extracts were washed with water, brine, dried,
and evaporated.
The residue was purified by recrystallization from EtzO.
1: 3.19g,(82.0%), pale brown solid.
1 H NMR (acetone-d6)
4.78 (s, 2H), 7.15 (t, J = 4.8Hz, 1H), 7.88-790 (m, 2H), 9.98 (brs,lH)
MS; 195(M+1)
Reaction 2
O N ~ N02 O N ~ NH2
O O2
1
A suspension of 1 (3.88g, 20.Ommo1) and 5% Pd/C (200mg) in a mixture of THF
(20mL)
and MeOH (60mL) was stirred under Ha atmosphere for 2h. The mixture was
filtered through
celite and the filtrate was concentrated. The residue was purified by column
chromatography
(MeOH/CHCI3 1:20) to give the product.
2: 1.89g, (57.6%), gray solid.
1 H NMR (acetone-d6)
4.39 (s, 2H), 4.42 (brs, 2H), 6.25 (dd, J = 8.5, 2.5 Hz, 1H), 6.32 (d, J =
2.SHz, 1H), 6.66 (d, J = 8.5
Hz, 1H), 9.36 (brs,lH)
MS; 165(M+1)
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Reaction 3
COOPh
H H I
O N ~ NH2 O N ~ NH
~I ~I
O O
2 3
To a mixture of 2 (880.Omg, 5.36mmo1) and pyridine (SlOmg, 6.43mmo1) in THF
(30mL)
was added chloro phenylcarbamate (840.Omg, 5.36mmo1) in THF (SmL) at
0°C. After l.Sh at rt,
the mixture was poured into water and extracted with EtOAc. The combined
extracts were washed
with brine, dried, and evaporated. The residue was purified by column
chromatography
(EtOAc/Hex 1l4) to give the product.
3: 1.255g, (82.4%) pale yellow solid.
1H NMR (acetone-d6)
4.53 (s,2H), 6.91 (d, J = 8.7Hz, 1H), 7.08 (d, J = 8.7Hz,lH), 7.19-7.24 (m,
3H), 7.39-7.43 (m, 3H),
9.10 (brs, l H), 9.71 (brs, l H).
MS; 285(M+1)
Reaction 4
I
H COOPh H3C O N~/'N~CH
O N ~ I NH H NON ~ CH3 O N , NH H CJ
o~ t 2 I I 3
O
3 4
A mixture of 3 (142mg, O.SOmmol), amine (89mg, O.SOmmol) and K2C03 (69mg,
O.SOmmol) in DMSO (SmL) was stirred for 15h at 80°C. After cooling, the
mixture was poured
into water and the precipitate was filtered, and then the solid was washed
with Et20 to obtain N-
f 2-[ethyl(3-methylphenyl)amino]ethyl}-N'-(3-oxo-3,4-dihydro-2H-1,4-benzoxazin-
6-yl)urea.
4; 177.Omg (96.1 %), brown solid.
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1H NMR (DMSO-d6)
1.07 (t, J = 6.9 Hz, 3H), 2.21 (s, 3H), 3.20 (dd, J = 13.3, 5.7 Hz, 2H), 3.31
(dd, J = 13.3, 5.7Hz,
2H), 3.35, (q, J = 6.9 Hz, 2H), 4.47 (s, 2H), 6.10 (t, J = 6.0 Hz, IH), 6.39
(d, J = 7.3 Hz, IH), 6.55
(s, 1H), 6.56 (d, J = 6.0 Hz, 1H), 6.78-6.83 (m, 2H), 7.02 (dd, J = 9.1, 7.3
Hz, 1H), 7.18 (d, J = 2.2
Hz, 1 H), 8.49 (s, 1 H), 10.6 (s, 1 H).
MS; 369 (M+I).
Result in capsaicin-induced Ca2+ influx assay: 24nM.
[Example 1-2]
Similarly,N-{2-[ethyl(3-methylphenyl)amino]ethyl~ N'-(2-oxo-1,2,3,4-
tetrahydroquinolin-5-yl)-
urea was obtained.
1 H NMR (DMSO-d6)
1.07 (t, J = 7.9 Hz, 3H), 2.21 (s, 3H), 2.41(t, J=7.3Hz, 2H), 2.72 (t,
J=7.3Hz, 2H), 3.22 (dd, J =
13.9, 6.9 Hz, 2H), 3.31 (q, J = 7.9Hz, 2H), 3.35 (dd, J = 13.9, 6.9Hz, 2H),
6.39 (d, J=7.3Hz, 1H),
6.48 (brs, 1H), 6.54-6.56 (m, 3n), 7.01 (d, J=7.3Hz, 1H), 7.03 (d, J = 7.9 Hz,
1H), 7.33 (d,
J=8.2Hz, 1 H), 7.87 (s, 1 H), 10.00 (s, 1 H).
MS; 367 (M+1).
Result in capsaicin-induced Ca2+ influx assay: 56nM.
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[Example 2]
Similary, the compounds described in the tables below are synthesized.
O
R~
O N ~ N- _N' L J ri N-R2
H H H
X n -R1 -R2
CF3
N
0 1 H
O-CF3
CHI 1 H
N
/ _N N-CH
3
0 1 -CH~CH3
/CF3
\J~i
N
0 1 -CH~CH3
C F3
CI
CHZ 2 -CH3
\\~N
0 2 / -CHZCH3
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[Example 3J
Similary, the compounds described in the tables below are synthesized.
O R~
~ ~ I
HN_ _N' L J ri N-R2
H
X
O N
H
X n - R1 - R~
CI
0 3
CH3
/
0 2 -CZHS OCH3
/ /
CHz 2 - CH~CH3
N
H
CHz 3 -CZHS-OCH3
N
CHZ 2
