Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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GLYCINE TRANSPORT INHIBITORS
The present invention is concerned with the use of glycine transport
inhibiting 4,4-bis-
(4-fluorophenyl)butyl]-1-(piperazinyl and piperidinyl) derivatives for the
preparation of
medicaments for treating disorders of the central and peripheral nervous
system, in
particular psychoses, pain, epilepsy, neurodegenerative diseases (Alzheimer's
disease),
stroke, head trauma, multiple sclerosis and the like. The invention further
comprises
novel compounds, their preparation and their pharmaceutical forms.
4,4-bis(4-fluorophenyl)butyl]-1-(piperazinyl and piperidinyl) derivatives are
well-
known histamine and serotonine antagonists. These compounds, their activity
and
preparation were disclosed in EP-A-0,151,826 and GB-1,055,100.
The present invention is concerned with the use of glycine transport
inhibiting
compounds for the preparation of medicaments for treating disorders of the
central and
peripheral nervous system, said compounds having the formula
F
CH-(CHz)3 N~ X-L (
F
the N-oxides, the stereochemically isomeric forms and the pharmaceutically
acceptable
addition salts thereof, wherein
X represents CH or N;
L represents a radical of formula
Rt
N a~a~ 2
-B1--C I t (a) ~ ~ ~R3
N~ 4 ~ as -.N-C_N\ (b)
a H
O
~N~NH (c)
l \
(CHZ)" (CH2)",
A\ / o
-Alk-1C-R'° (d)
wherein n is 0 or 1;
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mis0orl;
AIk represents C,_6alkanediyl;
A represents N or CH;
B' represents CH2 or NH;
-a'=a2-a3=a4- represents a bivalent radical of formula
-CH=CH-CH=CH- (a-1); or
-N=CH-CH=CH- (a-2);
R' represents C,_4alkyl optionally substituted with Ci_4alkyloxy, pyridinyl,
aryl, arylcarbonyl, thienyl, furanyl, imidazo[1,2-a]pyridinyl, thiazolyl;
R2 represents hydrogen or aryl;
R3 represents hydrogen, Ci_6alkyl or C3_~cycloalkyl;
R4 represents thienyl, furanyl, arylamino or a radical of formula
RS N
cd_ O
0
H
wherein RS is hydrogen or aryl;
aryl represents phenyl optionally substituted with 1 or 2 substituents
selected from
C,_4alkyl, halo, hydroxy, C,_4alkyloxy.
The present invention also relates to a method of treating warm-blooded
animals
suffering from disorders of the central and peripheral nervous system, in
particular
psychoses, pain, epilepsy, neurodegenerative diseases (Alzheimer's disease),
stroke,
head trauma, multiple sclerosis and the like. Said method comprises the
administration
of a therapeutically effective amount of a compound of formula (I) or a N-
oxide form, a
pharmaceutically acceptable acid or base addition salt or a stereochemically
isomeric
form thereof in admixture with a pharmaceutical carrier.
As used in the foregoing definitions and hereinafter, halo is generic to
fluoro, chloro,
bromo and iodo; C3_~cycloalkyl is generic to cyclopropyl, cyclobutyl,
cyclopentyl,
cyclohexyl and cycloheptyl; Cl~alkyl defines straight and branched chain
saturated
hydrocarbon radicals having from 1 to 4 carbon atoms such as, for example,
methyl,
ethyl, propyl, butyl, 1-methylethyl, 2-methylpropyl, 2,2-dimethylethyl and the
like;
C1_6alkyl is meant to include C1_4alkyl and the higher homologues thereof
having 5 or
6 carbon atoms such as, for example, pentyl, 2-methylbutyl, hexyl, 2-
methylpentyl and
the like; C1_6alkanediyl defines bivalent straight and branched chain
saturated
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hydrocarbon radicals having from 1 to 6 carbon atoms such as, for example,
1,1-methanediyl, 1,2-ethanediyl, 1,3-propanediyl, 1,4-butanediyl, 1,5-
pentanediyl,
1,6-hexanediyl, 1,2-propanediyl, 2,3-butanediyl and the like.
The pharmaceutically acceptable addition salts as mentioned hereinabove are
meant to
comprise the therapeutically active non-toxic base and acid addition salt
forms which
the compounds of formula (I) are able to form. The acid addition salt form of
a
compound of formula (I) that occurs in its free form as a base can be obtained
by
treating said free base form with an appropriate acid such as an inorganic
acid, for
example, hydrohalic acid, e.g. hydrochloric or hydrobromic, sulfuric, nitric,
phosphoric
and the like acids; or an organic acid, such as, for example, acetic,
hydroxyacetic,
propanoic, lactic, pyruvic, oxalic, malonic, succinic, malefic, fumaric,
malic, tartaric,
citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic,
cyclamic,
salicylic, p-aminosalicylic, pamoic and the like acids.
The compounds of formula (I) containing acidic protons may be converted into
their
therapeutically active non-toxic base, i.e. metal or amine, addition salt
forms by
treatment with appropriate organic and inorganic bases. Appropriate base salt
forms
comprise, for example, the ammonium salts, the alkali and earth alkaline metal
salts,
e.g. the lithium, sodium, potassium, magnesium, calcium salts and the like,
salts with
organic bases, e.g. the benzathine, N-methyl-D-glucamine, hydrabamine salts,
and salts
with amino acids such as, for example, arginine, lysine and the like.
Conversely said salt forms can be converted into the free forms by treatment
with an
appropriate base or acid.
The term addition salt as used hereinabove also comprises the solvates which
the
compounds of formula (I) as well as the salts thereof, are able to form. Such
solvates
are for example hydrates, alcoholates and the like.
The N-oxide forms of the compounds of formula (I) are meant to comprise those
compounds of formula (I) wherein one or several nitrogen atoms are oxidized to
the
so-called N-oxide.
The term "stereochemically isomeric forms" as used hereinbefore and
hereinafter
defines all the possible stereoisomeric forms in which the compounds of
formula (I)
may occur. Unless otherwise mentioned or indicated, the chemical designation
of
compounds denotes the mixture, and in particular the racemic mixture, of all
possible
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stereochemically isomeric forms, said mixtures containing all diastereomers
and
enantiomers of the basic molecular structure. Stereochemically isomeric forms
of the
compounds of formula (I) and mixtures of such forms are obviously intended to
be
encompassed by formula (I).
In particular, the compounds of formula (I) and some of the intermediates
hereinafter
have at least one stereogenic center in their structure. This stereogenic
center may be
present in a R and a S configuration, said R and S notation is used in
correspondance
with the rules described in Pure Appl. Chem., 1976, 45, 11-30.
I0
Some of the compounds of formula (I) may also exist in their tautomeric forms.
Such
forms although not explicitly indicated in the above formula are intended to
be
included within the scope of the present invention.
15 Whenever used hereinafter, the term compounds of formula (I) is meant to
include also
the N-oxides, the pharmaceutically acceptable addition salts and all
stereoisomeric forms.
The present compounds of formula (I) are deemed novel provided that
- when X is CH; L is a radical of formula (a) wherein B' is -CH2- and R' is
pyridin-2-
20 ylmethyl, thien-2-ylmethyl, furan-2-ylmethyl, benzyl or 4-fluorobenzyl,
then
-a'=a2-a~=a4- is other than a -N=CH-CH=CH-; and
- when X is CH; L is a radical of formula (a) wherein B' is -CH2- and R' is
4-methoxyphenylmetyl or thiazol-4-ylmethyl, then -a'=a2-a3=a4- is other than a
-CH=CH-CH=CH-; and
25 - when X is N; L is a radical of formula (d) wherein Alk is 1,3-
propanediyl, then R4 is
other than phenylamino.
The present invention also relates to the novel compounds of formula (I) as
defined
hereinabove for use as a medicine.
An interesting group of compounds are those compounds of formula (I) wherein n
is 0,
m is 1; R' is C,_4alkyl optionally substituted with C,_4alkyloxy, arylcarbonyl
or
imidazo( 1,2-a]pyridinyl and R4 is thienyl, furanyl or a radical of formula (d-
1 ).
Preferred compounds are the compounds of formula (I) wherein L is a radical of
formula (a) or (b).
In general, the compounds of formula (I) can be prepared according to reaction
procedures described in EP-A-0,151,826 and GB-1,055,100, more in particular,
by
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reacting an intermediate of formula (II) wherein Wi is an appropriate leaving
group
such as, for example, a halogen, with an intermediate of formula (III).
F
F ~ ~ CH-(CH~)3"~'~ + H-- ~ -L --~ (I)
(II) (III)
Said reaction may be performed in a reaction-inert solvent such as, for
example,
methylisobutyl keton, N,N-dimethylacetamide or N,N-dimethylformamide, in the
presence of a suitable base such as, for example, sodium carbonate, sodium
bicarbonate
or triethylamine, and optionally in the presence of potassium iodide.
In this and the following preparations, the reaction products may be isolated
from the
reaction medium and, if necessary, further purified according to methodologies
generally known in the art such as, for example, extraction, crystallization,
distillation,
trituration and chromatography.
Alternatively, the compounds of formula (I) can be prepared by reductive
alkylation.
An intermediate of formula (IV) is then reacted with an intermediate of
formula (III) in
a reaction-inert solvent such as, for example, methanol, in the presence of a
reducing
agent such as, for example, hydrogen in the presence of a suitable catalyst,
e.g.
palladium on activated charcoal, Conveniently, thiophene is added to the
reaction
mixture.
F
F ~ ~ CH
-(CHz)~-C-H + H-' ~X-L ---~ (I)
(IV) (III)
The compounds of formula (I) wherein X is N, said compounds being represented
by
formula (I-a), may be prepared by reacting an intermediate of formula (V) with
an
intermediate of formula (VI) wherein W' is a suitable leaving group such as
for
example, a halogen.
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+ W~-L --
(V)
Said reaction may be performed in a reaction-inert solvent such as, for
example,
methylisobutyl keton, N,N-dimethylacetamide or N,N-dimethylformamide, in the
presence of a suitable base such as, for example, sodium carbonate, sodium
bicarbonate
or triethylamine, and optionally in the presence of potassium iodide.
The compounds of formula (I) wherein L is a radical of formula (b), said
compounds
being represented by formula (I-b), may be prepared by reacting an
intermediate of
formula (VII) with an isocyanato derivative of formula (VIII).
F
I
F ~ ~ CH--(CHZ)3 ~ -N-H + O=C=N-R3
(VII) (VIII)
F
Rz O H
F ~ ~ CH,(CH2)3 ~ -N-C-N-R3
(I-b)
Said reaction may be performed in a reaction-inert solvent such as, for
example,
diisopropylether.
The compounds of formula (I) can also be converted into each other following
art-
known procedures of functional group transformation.
The compounds of formula (I) may also be converted to the corresponding N-
oxide
forms following art-known procedures for converting a trivalent nitrogen into
its
N-oxide form. Said N-oxidation reaction may generally be carried out by
reacting the
starting material of formula (I) with 3-phenyl-2-(phenylsulfonyl)oxaziridine
or with an
appropriate organic or inorganic peroxide. Appropriate inorganic peroxides
comprise,
for example, hydrogen peroxide, alkali metal or earth alkaline metal
peroxides, e.g.
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sodium peroxide, potassium peroxide; appropriate organic peroxides may
comprise
peroxy acids such as, for example, benzenecarboperoxoic acid or halo
substituted
benzenecarboperoxoic acid, e.g. 3-chlorobenzenecarboperoxoic acid,
peroxoalkanoic
acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g. t-butyl
hydroperoxide. Suitable
solvents are, for example, water, lower alkanols, e.g. ethanol and the like,
hydro-
carbons, e.g. toluene, ketones, e.g. 2-butanone, halogenated hydrocarbons,
e.g.
dichloromethane, and mixtures of such solvents.
Some of the compounds of formula (I) and some of the intermediates in the
present in-
vention may contain an asymmetric carbon atom. Pure stereochemically isomeric
forms of said compounds and said intermediates can be obtained by the
application of
art-known procedures. For example, diastereoisomers can be separated by
physical
methods such as selective crystallization or chromatographic techniques, e.g.
counter
current distribution, liquid chromatography and the like methods. Enantiomers
can be
obtained from racemic mixtures by first converting said racemic mixtures with
suitable
resolving agents such as, for example, chiral acids, to mixtures of
diastereomeric salts
or compounds; then physically separating said mixtures of diastereomeric salts
or
compounds by, for example, selective crystallization or chromatographic
techniques,
e.g. liquid chromatography and the like methods; and finally converting said
separated
diastereomeric salts or compounds into the corresponding enantiomers. Pure
stereochemically isomeric forms may also be obtained from the pure
stereochemically
isomeric forms of the appropriate intermediates and starting materials,
provided that the
intervening reactions occur stereospecifically.
An alternative manner of separating the enantiomeric forms of the compounds of
formula (I) and intermediates involves liquid chromatography, in particular
liquid
chromatography using a chiral stationary phase.
Some of the intermediates and starting materials are known compounds and may
be
commercially available or may be prepared according to art-known procedures.
Glycine is an amino acid neurotransmitter in the central and peripheral
nervous system,
both at inhibitory and excitatory synapses. These distinct functions of
glycine are
mediated by two types of receptor, each of which is associated with a
different class of
glycine transporter. The inhibitory actions of glycine are mediated by glycine
receptors
that are sensitive to the convulsant alkaloid strychnine, and are therefore
referred to as
'strychnine-sensitive.' Strychnine-sensitive glycine receptors are found
predominantly
in the spinal cord and brainstem.
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_g_
Glycine functions in excitatory transmission by modulating the actions of
glutamate,
the major excitatory neurotransmitter in the nervous system (Johnson and
Ascher,
Nature, 325, 529-531 (1987); Fletcher et al., Glycine Transmission, (Otterson
and
Storm-Mathisen, eds., 1990), pp. 193-219). Specifically, glycine is an
obligatory co-
y agonist at the class of glutamate receptor termed N-methyl-D-aspartate
(NMDA)
receptor. NMDA receptors are widely distributed throughout the brain, with a
particularly high density in the cerebral cortex and hippocampal formation.
Transporters take up neurotransmitter from the synapse, thereby regulating the
concentration and term of neurotransmitter in the synapse, which together
determine the
magnitude of synaptic transmission. By preventing the spread of
neurotransmitter to
neighboring synapses, transporters maintain the fidelity of synaptic
transmission. Last,
by re-uptake of released transmitter into the presynaptic terminal,
transporters allow for
transmitter reutilization. Neurotransmitter transport is dependent on
extraceilular
sodium and the voltage difference across the membrane. Under specific
conditions, for
example during a seizure, transporters can function in reverse, releasing
neurotransmitter in a calciumindependent non-exocytotic manner (Attwell et
al.,
Neuron, 11, 401-407 (1993)). Modulation of neurotransmitter transporters thus
provides a means for modifying synaptic activity, which provides useful
therapy for the
treatment of disturbances of the central and peripheral nervous system.
Molecular cloning has revealed the existence of two classes of glycine
transporters,
termed GIyT-1 and GIyT-2. GIyT-1 is found predominantly in the forebrain, and
its
distribution corresponds to that of glutamatergic pathways and NMDA receptors
(Smith,
et al., Neuron. 8, 927-935 (1992)). At least three splice variants of GIyT-1
are known,
namely GIyT-la, GIyT-lb and GIyT-lc (Kim, et al., Molecular Pharmacolo~v, 45,
608-
617 (1994)), each of which displays a unique distribution in the brain and
peripheral
tissues. GIyT-2, in contrast, is found predominantly in the brainstem and
spinal cord,
and its distribution corresponds closely to that of strychnine-sensitive
giycine receptors
(Liu et al., J Biological Chemistry. , 268,. 22802-22808 ( 1993); Jursky and
Nelson,
Neurochemistry, 64, 10261033 (1995)). Thus, one can expect that by regulating
the
synaptic levels of glycine, GIyT-1 and GIyT-2 selectively modulate the
activity of
NMDA receptors and strychnine-sensitive glycine receptors, respectively.
Compounds that inhibit or activate glycine transporters would thus be expected
to alter
receptor function, and provide therapeutic benefits in a variety of disease
states. Thus,
inhibition of GIyT-2 could be used to diminish the activity of neurons having
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strychnine-sensitive glycine receptors via increasing synaptic levels of
glycine, and so
diminish the transmission of pain-related (i.e., nociceptive) information in
the spinal
cord, which has been shown to be mediated by these receptors. Yaksh, Pain, 37,
1 I1-123 (1989). Additionally, enhancing inhibitory glycinergic transmission
through
strychnine-sensitive glycine receptors in the spinal cord can be used to
decrease muscle
hyperactivity, which is useful in treating diseases or conditions associated
with
increased muscle contraction, such as spasticity, myoclonus, and epilepsy
(Truong et
al., Movement Disorders, 3 , 77-87 ( 1988); Becker, FASEB J, 4 2767-2774 (
I990)).
Spasticity that can be treated via modulation of glycine receptors is
associated with
epilepsy, stroke, head trauma, multiple sclerosis, spinal cord injury,
dystonia, and other
conditions of illness and injury of the nervous system.
NMDA receptors are involved in memory and learning (Rison and Stanton,
Neurosci.
Biobehav. Rev., 19, 533 552 (1995); Danysz at al., Behavioral Pharmacol., 6,
455-474
(1995)); and decreased function of NMDA-mediated neurotransmission appears to
contribute to the symptoms of schizophrenia (Olney and Farber, Archives
General
Psychiatry, 52, 998-1007 (1996). Thus, agents that inhibit GIyT-1 and thereby
increase
glycine activation of NMDA receptors can be used as novel antipsychotics and
anti-dementia agents, and to treat other diseases in which cognitive processes
are
impaired, such as attention deficit disorders and organic brain syndromes.
Conversely,
over-activation of NMDA receptors has been implicated in a number of disease
states,
in particular the neuronal death associated with stroke, head trauma and
possibly
neurodegenerative diseases, such as Alzheimer's disease, mufti-infarct
dementia, AIDS
dementia, Huntington's disease, Parkinson's disease, amyotrophic lateral
sclerosis or
other conditions in which neuronal cell death occurs. Coyle & Puttfarcken,
Science,
262, 689-695 (1993); Lipton and Rosenberg, New Engl. J. of Medicine, 330, 613-
622
(1993); Choi, Neuron 1, 623-634 (1988). Thus, pharmacological agents that
increase
the activity of GIyT-1 will result in decreased glycine-activation of NMDA
receptors,
which activity can be used to treat these and related disease states.
Similarly, drugs that
directly block the glycine site on the NMDA receptors can be used to treat
these and
related disease states.
For administration purposes, the subject compounds may be formulated into
various
pharmaceutical compositions comprising a pharmaceutically acceptable Garner
and, as
active ingredient, a therapeutically effective amount of a novel compound of
formula
(I). To prepare the pharmaceutical compositions of this invention, an
effective amount
of the particular compound, in addition salt or in free acid or base form, as
the active
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ingredient is combined in intimate admixture with a pharmaceutically
acceptable
Garner, which may take a wide variety of forms depending on the form of
preparation
desired for administration. These pharmaceutical compositions are desirably in
unitary
dosage form suitable, preferably, for administration orally, percutaneously,
or by
parenteral injection. For example, in preparing the compositions in oral
dosage form,
any of the usual pharmaceutical media may be employed, such as, for example,
water,
glycols, oils, alcohols and the like in the case of oral liquid preparations
such as
suspensions, syrups, elixirs and solutions; or solid Garners such as starches,
sugars,
kaolin, lubricants, binders, disintegrating agents and the like in the case of
powders,
pills, capsules and tablets. Because of their ease in administration, tablets
and capsules
represent the most advantageous oral dosage unit form, in which case solid
pharmaceutical carriers are obviously employed. For parenteral compositions,
the
carrier will usually comprise sterile water, at least in large part, though
other
ingredients, for example, to aid solubility, may be included. Injectable
solutions, for
example, may be prepared in which the Garner comprises saline solution,
glucose
solution or a mixture of saline and glucose solution. Injectable solutions
containing
compounds of formula (I) may be formulated in an oil for prolonged action.
Appropriate oils for this purpose are, for example, peanut oil, sesame oil,
cottonseed
oil, corn oil, soy bean oil, synthetic glycerol esters of long chain fatty
acids and
mixtures of these and other oils. Injectable suspensions may also be prepared
in which
case appropriate liquid Garners, suspending agents and the like may be
employed. In the
compositions suitable for percutaneous administration, the carrier optionally
comprises
a penetration enhancing agent and/or a suitable wettable agent, optionally
combined
with suitable additives of any nature in minor proportions, which additives do
not cause
any significant deleterious effects on the skin. Said additives may facilitate
the
administration to the skin and/or may be helpful for preparing the desired
compositions.
These compositions may be administered in various ways, e.g., as a transdermal
patch,
as a spot-on or as an ointment. Addition salts of (I) due to their increased
water
solubility over the corresponding free base or free acid form, are obviously
more
suitable in the preparation of aqueous compositions.
It is especially advantageous to formulate the aforementioned pharmaceutical
composi-
tions in dosage unit form for ease of administration and uniformity of dosage.
Dosage
unit form as used in the specification and claims herein refers to physically
discrete
units suitable as unitary dosages, each unit containing a predetermined
quantity of
active ingredient calculated to produce the desired therapeutic effect, in
association
with the required pharmaceutical carrier. Examples of such dosage unit forms
are
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tablets (including scored or coated tablets), capsules, pills, powder packets,
wafers,
injectable solutions or suspensions, teaspoonfuls, tablespoonfuls and the
like, and
segregated multiples thereof.
The following examples are intended to illustrate the present invention.
Experi mental ,part
Example A.l
A mixture of 1-chloro-4,4-bis(4-fluorophenyl)butane (5.6 g), 4-(1,2,3,4-
tetrahydro-2-
oxo-3-quinazolinyl)piperidine (3.5 g), sodium carbonate (6.36 g), a few
crystals of KI
in methylisobutyl keton (160 ml) was stirred and refluxed for 2 days. After
cooling
water was added (250 ml). The separated organic layer was dried, filtered and
the
solvent evaporated. The residue was recrystallized from methylisobutyl keton
(80 ml),
yielding 3g of 3-[1-[4,4-bis(4-fluoromethyl)butyl)-4-piperidinyl]-3,4-dihydro-
2(1H)-
quinazolinone; mp. 199-200.5 °C (compound 1).
Analogously were prepared
4-[2-[ 1-(4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]acetyl]-3,4-dihydro-3-
phenyl-
2(1H)-quinoxalinone ethanedioate(1:1); mp. 190.8°C (compound 2);
N-[ 1-[4,4-bi s(4-fl uorophenyl)butyl]-4-piperidinyl]-1-(imidazo[ 1,2-
a]pyridin-2-
ylmethyl)-1H-benzimidazol-2-amine; mp. 160.1°C (compound 3);
2-[[4-[4,4-bis(4-fluorophenyl)butyl]-1-piperazinyl]methyl]-3-(2-ethoxyethyl)-
3H-
imidazo[4,5-b]pyridine ethanedioate (1:2); mp. 173.2°C (compound 4);
3-[ 1-[4,4-bi s(4-fluoromethyl)butyl]-4-piperidinyl]-3,4-dihydro-pyrido(2,3-d]-
2( 1 H)-
pyrimidinone dihydrochloride; mp. 220-222°C (compound 5).
Example A.2
A mixture of 4-fluoro-'y-(4-fluorophenyl)benzenebutanal (2.6 g), 1-(4-
fluorophenyl)-3-
[2-(4-piperidinylmethyl)-1H-benzimidazol-1-yl]-1-propanone dihydrobromide
monohydrate (5.5 g), a solution of thiophene in ethanol 3% (1 g), potassium
acetate
(3 g) and methanol (200 ml) was hydrogenated at normal pressure and at
50°C with
palladium-on-charcoal catalyst 10% (2 g). After the calculated amount of
hydrogen
was taken up, the catalyst was filtered off and the filtrate was evaporated.
Water was
added to the oily residue and the whole was alkalized with ammonium hydroxide.
The
product was extracted with 4-methyl-2-pentanone. The extract was dried,
filtered and
evaporated. The oily residue was purified by column chromatography over silica
gel
using a mixture of trichloromethane and methanol (95:5 by volume) as eluent.
The oily
residue was converted into the ethanedioate salt in acetonitrile and 4-methyl-
2-
pentanone. The salt was allowed to crystallize. The product was filtered off
and dried,
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yielding 5 g (63.3 %) of 3-[2-[[1-[4,4-bis(4-fluorophenyl)butyl]-4-
piperidinyI]methyl]-
1H-benzimidazol-1-yl]-1-(4-fluorophenyl)-1-propanone ethanedioate (1:2);
mp. 156.4°C (compound 6).
Example A.3
A mixture of 1-[4,4-bis(4-fluorophenyl]butyl]-piperazine (6.9 g), 4-chloro-1-
(2-thienyl)-
butanone (4,1 g), sodium carbonate (3.18 g), a few crystals of potassium
iodide in
4-methyl-2-pentanone (200 ml) was refluxed for 24 hours. Then a second portion
of
4-chloro-1-(2-thienyl)butanone (4.1 g) was added and the whole was stirred and
refluxed
for an additional 36 hours. After cooling, water was added (100 ml). The
organic layer
was separated, dried over potassium carbonate, filtered and evaporated. The
oily residue
was dissolved in anhydrous ether (480 ml). The solution was filtered and
gaseous
hydrogen chloride was introduced into the filtrate. The precipitated salt was
filtered off
and crystallized from 2-propanol (320 ml), yielding 4-[4,4-bis(4-
fluorophenyl)butyl]-1-
piperazinyl]-1-(2-thienyl)butanone; mp. 227.5-230°C (compound 7).
Example A.4
To a stirred solution of 1-[4,4-bis(4-fluorophenyl)butyl]-N-(4-methoxyphenyl)-
4-
piperidinamine (6.75g) in 2,2'-oxybispropane (105 mI) and tetrahydrofuran (45
ml) was
added dropwise a solution of 2-isocyanatopropane (1.36 g) in 2,2'-
oxybispropane
(35 ml). Upon completion, stirnng was continued first for 1 day at room
temperature
and further for 1 hour at about 50°C. The reaction mixture was
evaporated and the
residue was crystallized from a mixture of 2,2'-oxybispropane and 2-propanol,
yielding
N-[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]-N-(4-methoxyphenyl)-N'-(1-
methyl-
ethyl)urea (4.8 g, 59%); mp. 170.9°C (compound 8).
Analogously were prepared
N-[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]-N'-butyl-N-(4-
methoxyphenyl)urea;
mp. 101.9°C (compound 9);
N-[ 1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]-N-(4-methoxyphenyl)-N'-
propyl-
urea; mp. 124.1 °C (compound 10);
N-[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]-N'-cyclohexyl-N-(4-methoxy-
phenyl)urea; mp. 128.2°C (compound 11);
N-[ 1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl)-N'-ethyl-N-(4-
methylphenyl)urea;
mp. 129.1 °C (compound 12);
N-[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]-N'-( 1-methylethyl)-N-(4-
methyl-
phenyl)urea; mp. 167.2°C (compound 13); and
N-[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]-N'-(4-chlorophenyl)-N'-(1-
methyl-
ethyl)urea; mp. 157.4°C (compound 14).
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WO 99/44596 PCTlEP99/01309
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Phanmacolo~ical example
Example B.1: Assay of transport via GlyTl transporters
Subconfluent HEK 293 -GlyT1 cells (i.e. a cell line which stably expresses
human
glycine transporter 1 ) were seeded in Cytostar-T plates at a concentration of
50,000
cells per well in 100 pl DMEM medium (Dulbecco's Modified Eagle Medium
supplemented with 10% foetal bovine serum, 1 mM Na-pyruvate, 2 mM glutamine,
100 U penicillin/ml and 0.1 mg/ml streptomycin). The cells were incubated for
48
hours at 37°C, 5% C02, 95% humidity.
On day 3> the cells were washed using a Tecan PW96 microprocessor controlled
washer
designed to wash all 96 wells of a microplate simultaneously with uptake
buffer
(25 mM Hepes, 5.4 mM K-gluconate, 1.8 mM Ca-gluconate, 0.8 mM MgS04, 140 mM
NaCI, 5 mM glucose, 5 mM alanine, adjusted to pH 7.5 with 2M Tris). The Tecan
PW96 was programmed to wash the cells five times leaving 75 pl in each well.
The
test compounds were dissolved at different concentrations in the micromolar
range in
DMSO. 1 pl Test solution was added to each well and the cells were incubated
for 5'
to 10' at ambient temperature. Then there was added 25 p,l 30 ~M [U'4C]glycine
diluted in uptake buffer. The cells were incubated for 1 hour at ambient
temperature.
The plates were then sealed and [U'4C]glycine uptake was determined on a
Packard
microplate scintillation counter (TopCount). From the results obtained for the
various
concentrations of each test drug, the concentration giving 50 % inhibition
(ICso) of
glycine uptake was calculated. Calculated data for the test compounds
according to the
instant invention are shown in table I as pICso values (negative log values of
the ICSO).
Comp. 15 being 2-[[I-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-(2-
pyridinylmethyl)-3H-imidazo[4,5-b]pyridine ethanedioate (1:2);
comp. 16 being 2-[[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-
[(4-fluor-
phenyl)methyl]-3H-imidazo[4,5-b]pyridine ethanedioate (1:2);
comp. 17 being 2-[[I-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-
(phenyl-
methyl)-3H-imidazo[4,5-b]pyridine ethanedioate (I:2);
comp. 18 being 2-[[I-(4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-(2-
thienyl-
methyl)-3H-imidazo[4,5-b]pyridine ethanedioate (1:1);
comp. 19 being 2-[[I-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-
(2-furanylmethyl)-3H-imidazo[4,5-b]pyridine ethanedioate (1:2);
comp. 20 being 2-[[I-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-
[(4-methoxy-phenyl)methyl]-1H-benzimidazole ethanedioate (1:2);
comp. 21 being 2-[[1-[4,4-bis(4-fluorophenyl)butyl]-4-piperidinyl]methyl]-3-
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(4-thiazolyl-methyl)-1H-benzimidazole ethanedioate (1:2); as disclosed in
EP-A-0,151,826;
and comp. 22 being 1-[4,4-di(4-fluorophenyl)butyl]-4-(3-(anilinocarbonyl}-
propyl]-
piperazine dihydrochloride as disclosed in GB-1,055,100, were also tested.
Table 1
Com No. ICso Com No. ICso Com No. ICso
1 6.61 9 6.15 17
6.42
2 6.34 10 6.12 18
6.26
3 6.14 11 6.28 19
6.15
4 6.81 12 6.02 20
6.09
5 6.26 13 6.18 21
6.07
6 6.44 14 6.18 22
6.33
7 6.29 15 6.65
8 6.51 16 6.55
C. Composition examples
The following formulations exemplify typical pharmaceutical compositions
suitable for
systemic administration to animal and human subjects in accordance with the
present
invention.
"Active ingredient" (A.L) as used throughout these examples relates to a
compound of
formula (I) or a pharmaceutically acceptable addition salt thereof.
Example C.1 : film-coated tablets
Preparation of tablet core.A mixture of 100 g of the A.L, 570 g lactose and
200. ...... g starch
was mixed well and thereafter humidified with a solution of S g sodium dodecyl
sulfate
and 10 g polyvinylpyrrolidone in about 200 ml of water. The wet powder mixture
was
sieved, dried and sieved again. Then there was added 100 g microcrystalline
cellulose
and 15 g hydrogenated vegetable oil. The whole was mixed well and compressed
into
tablets, giving 10.000 tablets, each comprising 10 mg of the active
ingredient.
Coating To a solution of 10 g methyl cellulose in 75 ml of denaturated ethanol
there
was added a solution of 5 g of ethyl cellulose in 150 ml of dichloromethane.
Then there
were added 75 ml of dichloromethane and 2.5 ml 1,2,3-propanetriol. 10 g of
polyethylene glycol was molten and dissolved in 75 ml of dichloromethane. The
latter
solution was added to the former and then there were added 2.5 g of magnesium
octadecanoate, 5 g of polyvinylpyrrolidone and 30 ml of concentrated color
suspension
and the whole was homogenated. The tablet cores were coated with the thus
obtained
mixture in a coating apparatus.
