Title of Invention

"A SUBSTITUTED QUINOLONE AND COMPOSITION COMPRISING THE SAME"

Abstract A substituted quinolone having the Formula I or a pharmaceutically acceptable salt, amide, ester or solvate thereof, wherein: R1 is selected from the group consisting of hydrogen; an optionally substituted C1-20 alkyl, and C7-30 aralkyl; each R2 is selected from the group consisting of hydrogen and optionally substituted C1-20 alkyl; each R3 is selected from the group consisting of hydrogen, optionally substituted C1-20 alkyl; a group OR11 and NR12R13; R5, R7 and Rs are independently selected from the group consisting of hydrogen, an optionally substituted C1-20 alkyl, and halogen; R9 and R1o are independently selected from the group consisting of hydrogen, optionally substituted C1-20 alkyl, C7-30 aralkyl, C3-8 cycloalkyl and C6-22 cycloaralkyl; with the proviso that R9 and R10 are not both hydrogen at the same time; R11 is selected from the group consisting of hydrogen, an alkali metal, a negative charge and optionally substituted C1-20 alkyl; R12 and R13 are independently selected from the group consisting of hydrogen, optionally substituted C1-20 alkyl, C7-30 aralkyl, C6-14- aryl, C3-8 cycloalkyl and C6-22 cycloaralkyl; or R12 and R13 are taken together with the nitrogen atom to which they are attached to form a heterocyclic ring.
Full Text The present invention relates to a substituted quinolone and composition comprising the same.
Related Application
This application claims priority to U.S. provisional application serial number 60/380,641, filed on May 14,2002.
Field of the Invention
This invention is in the field of medicinal chemistry. In particular, the invention relates to substituted quinolone carboxylic acids and their derivatives, which modulate, via a unique site, the effect of γ-aminobutyric acid (GABA) on the GABAA receptor complex in a therapeutically relevant fashion and may be used to ameliorate CNS disorders amenable to modulation of the GABAA receptor complex.
Background of the Invention
GABA is the most abundant inhibitory neurotransmitter in the mammalian brain. GABA controls brain excitability by exerting inhibitory functions on neuronal membranes by altering their permeability to specific ions. Binding of GABA to the GABAA-type (GABAA) receptor increases the permeability of neuronal membranes to chloride ions (C1-). In most neurons the relative C1- ion concentration is greater outside than the inside the membrane. Thus, selective permeability to CI- initiated by GABA binding allows C1- to flow down its electrochemical gradient into the cell. The majority of fast inhibitory synaptic transmission is a result of GABA binding to the GABAA receptors. GABAA receptors are ubiquitously expressed throughout the CNS with almost all neurons staining for their presence. The GABAA receptor is a hetero-pentameric protein structure of the nicotinic acetylcholine receptor superfamily. Native GABAA receptors are formed from at least 19 related subunits. The subunits are grouped into α,β,δ,ε, ,and p families. The most prevalent combination of GABAA receptors is a
stoichiometnc combination of the 2 x a, 2 x P, and 1 x y subunits, with the remaining
subunits relegated to substituting for the j subunit during specific development
expression or in highly specific brain region localization. The adult brain predominately
express the alp2v2 subunit combination (60%) with the o2p3v2 and a3pny2 subunits
comprising the majority (35%) of the remaining receptors. The relative effects of GAB A
are influenced by the GABAA receptor subunit expressed in a specific brain region or
neuronal circuit.
The neurophysiological effects of GAB A result from a conformational change
that occurs when GABA binds to the GABAA receptor. The GABAA receptor and the
associated ion channel complex (GRC) is a h'gand-gated ion channel which recognizes
many compounds that allosterically modulate the ability of GABA to bind to the GABAA
receptor. The allosteric modulators have distinct sites on the GRC. These sites are
separate and unique from the site that recognizes GABA. The most widely studied and
characterized class of allosteric modulator of the GRC is that which interact with the
benzodiazepine (BZ)-site.
Alternative sites for modulating the GRC have been described. For example,
neuroactive steroids are non-hormonal steroids that bind and functionally modulate the
GRC. The current role of neuroactive steroids in GABAA receptor pharmacology is
supported by overwhelming evidence. Electrophysiological and biochemical techniques
have confirmed the capacity of neuroactive steroids to allosterically modulate the GRC
through a unique site of action. Experimentally neuroactive steroids exhibit a
pharmacological'profile similar, but not identical, to the benzodiazepines. Neuroactive
steroids produce anxiolytic, anticonvulsant, and sedative-hypnotic properties.
Certain antibacterial fluoroquinolone antibiotics have been implicated in clinical
reports as the cause of convulsions in humans (Ball P (1986) Journal of Antimicrobial
Chemotherapy. 18 SupplD 187-193; Simpson KJ, BrodieMJ( 1985) Lancet ii: 161,
1985; Hori S, et al. (1987) Program and Abstracts of the Twenty-Seventh Interscience
Conference on Antimicrobial Agents and Chemotherapy, New York 1987. Abstract 30,
pg 101). Experimentally, fluoroquinolones have been demonstrated to produce
convulsions and death in mice. Additionally, non-steroidal anti-inflammatory drugs
(NSAIDs) and their by-products have been reported to clinically and experimentally
potentiate the convulsive effects of the fluoroquinolones. Concerns about the convulsant
side-effects of fluoroquinolone antibacterial agents have led to an interest in the
interaction of fluoroquinolones with the GABAA receptor. Convincing evidence has
accumulated that suggests that they interact with the GRC to inhibit GABA action.
Fluoroquinolones antagonize [3H]muscimol and [3H]GABA binding to the GRC with
high micromolar potency. Electrophysiological studies have demonstrated that
fluoroquinoiones alone weakly reduce GABA-evoked currents. As well, radioligand
binding assays have shown that fluoroquinolones, in combination with NSAIDs, induce a
conformational change in the GABAA receptor-chloride channel complex that is
indicative of a pharmacologically relevant response consistent with functional
antagonism of GABA.
It is well-documented that modulation of the GRC can ameliorate anxiety, seizure
activity, and insomnia. Thus, GABA and drugs that act like GABA or facilitate the
effects of GABA (e.g., the therapeutically useful barbiturates and benzodiazepines (BZs)
such as Valium) produce their therapeutically useful effects by interacting with specific
modulatory sites on the GRC. None of the known drugs, however, are selectively potent
at the a-2 subunit of the GABA receptor. Thus, they exhibit undesirable side effects of
sedation, and in the case of fluoroquinolones, convulsions. There is presently a need for
GRC modulators that are active without side effects.
Summary of the Invention
The present invention relates to molecules that modulate the GRC with selective
potency at the a-2 subunit of GABA to produce therapeutically useful effects without
side effects. The present invention further relates to substituted quinolones represented
by Formula I that act as enhancers of GABA-facilitated Cl'flux mediated through the
GABAA receptor complex (GRC).
The invention also relates to methods of treating disorders responsive to
enhancement of GABA action on GABA receptors in a mammal by administering an
effective amount of a compound of Formula I and by activation of the novel site which
mediates the action of a compound of Formula I as described herein. The novel site is
defined by exclusion criteria where a compound of Formula I does not act on known
sites of the GRC which include the sites for GABA, benzodiazepines, neuroactive
steroids, t-butylbicyclophosphorothionate/picrotoxin, barbiturates, 4'-chlorodiazepam,
antibacterial quinolones, ivermectin, loreclezole/mefanamic acid, furosemide and
propofol (E.R. Korpi, G. Grunder, H. Luddens, Progress Neurobiology 67:113-159,
2002).
The compounds of the present invention, being ligands for a unique site on the
GRC, are therefore of use in the treatment and/or prevention of a variety of disorders of
the central nervous system. Such disorders include anxiety disorders, such as panic
disorder with or without agoraphobia, agoraphobia without history of panic disorder,
animal and other phobias including social phobias, obsessive-compulsive disorder, stress
disorders including post-traumatic and acute stress disorder, and generalized or
substance-induced anxiety disorder, neuroses; convulsions; acute and chronic pain;
cognitive disorders; insomnia; migraine; and depressive or bipolar disorders, for example
single-episode or recurrent major depressive disorder, dysthymic disorder, bipolar I and
bipolar II manic disorders, and cyclothymic disorder.
Another aspect of the present invention is directed to the use of the site that
mediates the activity of compounds of Formula I as enhancers or inhibitors of GAB Afacilitated
Cl" conductance mediated through the GABAA receptor complex.
Enhancement of GABA-rnediated chloride conductance is useful for the treatment and
prevention of such disorders as anxiety and stress related disorders, depression and other
affective disorders, epilepsy and other seizure disorders, insomnia and related sleep
disorders, and acute and chronic pain. Inhibition of GABA-mediated chloride
conductance is useful for the treatment and prevention of disorders related to learning
and memory such as mild cognitive impairment, age related cognitive decline, senile
dementia, Alzhiemer's disease, sleep disorders involving reduced wakefulness such as
narcolepsy and idiopathic hypersomnia.
Also, an aspect of the present invention is to provide a pharmaceutical
composition useful for treating disorders responsive to the enhancement GABAfacilitated
Cl" flux mediated through the GRC, containing an effective amount of a
compound of Formula I in a mixture with one or more pharmaceutically acceptable
carriers or diluents.
Compounds useful in the present invention have not been heretofore reported.
Thus, the present invention is also directed to novel substituted quinolones having the
structure of Formula I.
Further, the present invention is directed to 3H, 35S, 36C1,125I, 131I and 14C
radiolabeled compounds of Formula I and their use as a radioligand for their binding site
on the GRC.
Additional embodiments and advantages of the invention will be set forth in part
in the description that follows, and in part will be obvious from the description, or may
be learned by practice of the invention. The embodiments and advantages of the
invention will be realized and attained by means of the elements and combinations
particularly pointed out in the appended claims.
It is to be understood that both the foregoing general description and the
following detailed description are exemplary and explanatory only and are not restrictive
of the invention, as claimed.
Brief Description of the Drawings
Fig. 1 depicts the potentiating effect of 7-chloro-l-ethyl-6-(l,2,3,4-
tetrahydronaphthyl-l-amino)-4-oxo-l,4-dihydroquinoline-3-carboxylic acid (C3, S^M)
on GABA (G, 10 ^M) induced chloride currents in embryonic rat hippocampal neurons.
These data demonstrate that C3 is a positive efficacy modulator of GAB A-gated chloride
channels.
Fig. 2 depicts receptor subunit selectivity and dose-dependent positive efficacy of
7-chloro-l-ethyl-6-(l,2,3,4-tetrahydro-naphthyl-l-amino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid (compound 3, CMP 3) versus diazepam (DZP) on GABA induced
currents (loABA)in expressed human GABAA receptors containing CL^2f2 versus o^pVh
subunits.
Fig. 3 depicts a comparison of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-] -
amino)-4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3) and Diazepam
(DZP) on time spent in the dark in the Mouse Light-Dark Transition Model of Anxiety.
These data demonstrate that the anti-anxiety effects, as shown by the increase in the rime
spent in the dark, of compound 3 are comparable to that of DZP.
Fig. 4 depicts a comparison of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l,4-dihydroquinoline-3-carboxylic acid (CMP 3) and Diazepam (DZP) on
punished responding as measured by the number of licks during a 3 minute period in the
Vogel Model of Anxiety using 24 hour thirsted rats. These data demonstrate that the
anti-anxiety effects, as shown by increased punished licking, of compound 3 are
comparable to that of DZP.
Fig. 5 depicts an effect of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l14-dihydroquinoline-3-carboxylic acid (compound 3) on 2 nM [ S]TBPS
binding to rat cortex in the absence (open circles) or presence or of 3 uM (closed circle)
and 10 nM (closed square) of the GABAA receptor antagonist (+)-bicuculline. These
data demonstrate the absolute dependence of compound 3 on GAB A for efficacy and that
compound 3 is allosterically coupled to and does not act directly on the [ 5S]TBPS site.
Fig. 6 depicts an effect of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3, closed circle),
clonazepam (open circle) and 5ct-pregnan-3a-ol-20-one (3a,5a-P, open square) on 0.2
nM [3H}flunitrazepam binding to BZ receptors in rat cortex. These data demonstrate
that compound 3 is allosterically coupled to and does not act directly on the BZ receptor.
Fig. 7 depicts an effect of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3, closed circle) and
GABA (open circle) on 5 nM [3H]muscimol binding to the GABAA receptor in rat
cortex. These data demonstrate that compound 3 does not act directly on the GABAA
receptor.
Fig. 8 depicts an effect of 10 (aM 7-chloro-l-ethyl-6-( 1,2,3,4-tetrahydronaphthyll-
amino)-4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3, closed circle) or
100 nM 3a,5a-P (open square) on 5a-pregnan-3a,20a-diol (5a,20a-diol, open circle)
inhibition of 2 nM [33S]TBPS binding to rat cortex. As predicted, increasing
concentrations of 5a,20oc-diol (a partial agonist) antagonize the effect of 3a,5cc-P (a full
agonist). The inability of 5a,20a-diol to antagonize the effect of compound 3
demonstrates that compound 3 does not act directly on the neurosteroid site of the GRC.
Fig. 9 depicts the effect of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3) on 2 nM [35S]TBPS
binding to rat cortex in the absence (open circle) or presence of 30 nM norfloxacin
(closed circle) and 100 |o.M norfloxacin (closed square). The inability of norfloxacin to
produce a dose-dependent rightward parallel shift of the compound 3 dose-response
demonstrates that compound 3 does not act directly at the same site as the antibacterial
quinolone norfloxacin.
Fig 10 depicts the dissociation of 2 nM [3SS]TBPS binding from rat cortex
initiated by 10 joM 7-chloro-l-ethyl-6-(l,2,3,4-terrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid (compound 3) in the absence (closed square) or
presence (closed triangle) of 30 uM pentobarbital. The ability of pentobarbital to
accelerate the dissociation of [35S]TBPS binding indicates that compound 3 and the
barbiturate pentobarbital do not share a common site of action.
Fig, 11 depicts the effect of 7-chloro-l-ethyl-6-(l>2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3) on 2 nM [35S]TBPS
binding to rat cortex in the absence (open circle) or presence of 0.3 )aM (closed circle), 1
jiM (closed square) and 30 uM Ro5-4864 (4'-chlorodiazepam, closed triangle). The
inability of 4'-chlorodiazepam to produce a dose-dependent rightward parallel shift of
the compound 3/[35S] TBPS dose-response curve demonstrates that compound 3 does not
act directly at the same site as 4'-chlorodiazepam.
Fig 12 depicts the dissociation of 2 nM [35S]TBPS binding from mouse forebrain
initiated by 10 p.M 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid (compound 3) in the absence (open circle) or
presence (closed square) of 10 |iM ivermectin. The ability of ivermectin to accelerate
the dissociation of [35S]TBPS binding indicates that compound 3 and ivermectin do not
share a common site of action.
Fig 13 depicts the dissociation of 2 nM [35S]TBPS binding from mouse forebrain
initiated by 10 p.M 7-chloro-l-ethyl-6-(l,2)3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid (compound 3) in the absence (open circle) or
presence (closed square) of 10 jaM mefenamic acid. The ability of mefenamic acid to
accelerate the dissociation of [3SS]TBPS binding indicates that compound 3 and
mefenamic acid do not share a common site of action.
Fig. 14 depicts the effect of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-lamino)-
4-oxo-l,4-dihydroquinoline-3-carboxylic acid (compound 3) on 2 nM [35S]TBPS
binding to rat cerebellum in the absence (open circle) or presence of 30 joM furosemide
(closed circle). The inability of furosemide to produce a dose-dependent rightward
parallel shift of the compound 3 dose-response demonstrates that compound 3 does not
act directly at the same site on the GRC as the loop-diuretic ftrrosemide.
Detailed Description of the Invention
The compounds useful in this aspect of the invention are substituted quinolones
represented by Formula I:
or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein:
Ri is selected from the group consisting of hydrogen; an optionally substituted
alkyl, amino, aryl and aralkyl;
each Ra is selected from the group consisting of hydrogen and optionally
substituted alkyl;
each Rj is selected from the group consisting of hydrogen, optionally substituted
alkyl; a group ORu and NRnRn;
RS, R? and Rg are independently selected from the group consisting of hydrogen,
an optionally substituted alkyl, and halogen;
Rg and RIO are independently selected from the group consisting of hydrogen,
optionally substituted alkyl, aralkyl, cycloalkyl and cycloaralkyl; or Rg and RIO are taken
together with the nitrogen atom to which they are attached to form a heterocyclic ring
with the proviso that Rj and RIO are not both hydrogen at the same time;
RI i is selected from the group consisting of hydrogen, an alkali metal, a negative
charge and optionally substituted alkyl;
RU and RU are independently selected from the group consisting of hydrogen,
optionally substituted alkyl, aralkyl, aryl, cycloalkyl and cycloaralkyl; or Ria and Ria are
taken together with the nitrogen atom to which they are attached to form a heterocyclic
ring.
The invention also relates to quinolones represented by Formula II:
(Figure Removed)
or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein:
RI is selected from the group consisting of hydrogen; an optionally substituted
alkyl, and aralkyl;
each RI is selected from the group consisting of hydrogen and optionally
substituted alkyl;
RS, R7 and Rg are independently selected from the group consisting of hydrogen,
an optionally substituted alkyl, and halogen;
Rg and RIO are independently selected from the group consisting of optionally
substituted alkyl, aralkyl, cycloalkyl and cycloaralkyl; or Rg and RIO are taken together
with the nitrogen atom to which they are attached to form a heterocyclic ring.
Also, the invention relates to compounds of Formula
(Figure Removed)
or a pharmaceutically acceptable salt, prodrug or solvate thereof, wherein:
RI, R2, RS, R?, Ra, Rg are defined previously with respect to Formulae I and
(Figure Removed)and n is an integer 0, 1,2, 3 or 4.
For use in medicine, the salts of the compounds of Formula
(Figure Removed)will be
pharmaceutically acceptable salts. Other salts may, however, be useful in the preparation
of the compounds according to the invention or of their pharmaceutically acceptable
salts. Suitable pharmaceutically acceptable salts of the compounds of this invention
include acid addition salts which may, for example, be formed by mixing a solution of
the compound according to the invention with a solution of a pharmaceutically
acceptable acid such as hydrochloric acid, sulruric acid, methanesulfom'c acid, fumaric
acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric
acid, or phosphoric acid. Furthermore, where the compounds of the invention carry an
acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali
metal salts, e.g. sodium or potassium salts; alkaline earth metal salts, e.g. calcium or
magnesium salts; and salts formed with suitable organic ligands, e.g. quaternary
ammonium salts.
The present invention includes within its scope prodrugs of the compounds of
Formula I above. In general, such prodrugs will be functional derivatives of the
compounds of Formula I which are readily convertible in vivo into the required
compound of Formula I. Conventional procedures for the selection and preparation of
suitable prodrug derivatives are described, for example, in Design of Prodrugs, ed. H.
Bundgaard, Elsevier, 1985.
Where the compounds according to the invention have at least one asymmetric
center, they may accordingly exist as enantiomers. Where the compounds according to
the invention possess two or more asymmetric centers, they may additionally exist as
diastereoisomers. It is to be understood that all such isomers and mixtures thereof in any
proportion are encompassed within the scope of the present invention.
Useful halogen groups include fluorine, chlorine, bromine and iodine.
Useful alkyl groups include straight chain and branched Cl-20 alkyl groups,
more preferably, C5-20 alkyl groups. Typical C5-20 alkyl groups include n-penyl, nhexyl,
n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tricedyl, n-tetradecyl,
n-pentadecyl, n-hexadecyl, n-heptadecyl, n-octadecyl, n-nonadecyl and eicosanyl groups.
Useftil aryl groups are CS-H aryl, especially Cg-io aryl. Typical Ce-u aryl groups
include phenyl, naphthyl, anthracyl, indenyl, and biphenyl groups.
Useful arylalkyl groups include any of the above-mentioned Cl-20 alkyl groups
substituted with any of the above-mentioned C6-10 aryl groups. Useful arylalkyl groups
include benzyl and phenethyl.
Useful cycloalkylalkyl groups include any of the above-mentioned Cl-20 alkyl
groups substituted with any of the previously mentioned cycloalkyl groups. Examples of
useful cycloalkylalkyl groups include cyclohexylmethyl and cyclopropylmethyl groups.
Useful halomethyl groups include Cl-20 alkyl groups substituted with one or
more fluorine, chlorine, bromine or iodine atoms, including fluoromethyl,
difluoromethyl, trifluoromethyl and 1,1-difluoroethyl groups.
Useful hydroxyalkyl groups include Cl-20 alkyl groups substituted by hydroxy,
including hydroxymethyl, 1- and 2-hydroxyethyl and 1-hydroxypropyl groups.
Useful alkoxy groups include oxygen substitution by one of the Cl-20 alkyl
groups described above.
Useful alkylthio groups include sulfur substitution by one of the Cl-20 alkyl
groups described above including decyl- and hexadecylthio groups.
Useful alkylamino and dialkylamino are -NHRg and -NRgRio, wherein Rg and
RIO are Cl-20 alkyl groups.
Useful dialkylaminoalkyl groups include any of the above-mentioned Cl-20 alkyl
groups substituted by any of the previously mentioned dialkylamino groups.
Useful alkylthiol groups include any of the above-mentioned Cl-20 alkyl groups
substituted by a -SH group.
A carboxy group is -COOH.
An amino group is -NHj.
The term heterocyclic is used herein to mean saturated or wholly or partially
unsaturated 3-7 membered monocyclic, or 7-10 membered bicyclic ring system, which
consists of carbon atoms and from one to four heteroatoms independently selected from
the group consisting of 0, N, and S, wherein the nitrogen and sulfur heteroatoms can be
optionally oxidized, the nitrogen can be optionally quaternized, and including any
bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene
ring, and wherein the heterocyclic ring can be substituted on carbon or nitrogen if the
resulting compound is stable. Examples include, but are not limited to pyrrolidine,
piperidine, piperazine, morpholine, 1,2,3,4-tetrahydroquinoline, and the like.
Optional substituents on RI to Ri3 include any one of halo, halo(Ci-2o)alkyl, aryl,
cycloalkyl, Ci-aoalkyl, aryl(C|.2o)alkyl, cycloalkyl(C|.2o)alkyl, hydroxy(Ci-2o)alkyl,
amino(Ci-2o)alkyl, alkoxy(Ci-2o)alkyl, amino, hydroxy, thiol, alkoxy, and C1-20 alkylthiol
groups mentioned above. Preferred optional substituents include: halo, halo(C|.6)alkyl,
amino(Ci-6)alkyl, alkoxy and amino.
The synthesis of compounds of Formula I where R? = Cl and RIO = H can be
accomplished by reacting a primary amine, RgNEfe, in l-methyl-2-pyrrolidinone (NMP)
with 7-chloro-l-ethyl-6-fluoro-4-oxo-l,4-dihydroquinoline-3-carboxylic acid (2,
(Figure Removed)
substituted benzylamines, substituted phenethylamines, 3-phenylaminopropane, 1-
aminoindan and l-ammo-l.l.S^-tetrahydronaphthlene.
For the synthesis of compounds of Formula I with groups other than ethyl and
cyclopropyl at RI, the 6-fluoro-7-chloro starting material (8) can be prepared as in
Scheme 2 starting from commercially available 2,4-dichloro-5-fluorobenzoyl chloride (4,
Lancaster Synthesis).
(Figure Removed)
Examples of RiNKb include 2-fluoroethylamine, optionally substituted
benzylamines and optionally substituted phenethylamines. Other methods for
assembling the quinolone ring can be used as described in Atkins, et al, Org. Process
Res. & Develop. (1997), 1, 185-197.
In Vitro Binding Assay 1
^SJTBPS binding assay. The cortex from male Sprague-Dawley rats (weighing 160-
200g) was removed immediately after decapitation and dissected over ice. A Pa
homogenate was prepared for binding assay as previously described (Gee KW
Phenylquinolines PK 8165 and PK 9084 allosterically modulate [35S]tbutylbicyclophosphorothionate
binding to a chloride ionophore in rat brain via a novel
Ro5 4864 site. J. Pharmacol. Exp. Ther. 240:747-753, 1987). The tissue was
homogenized in 0.32M sucrose (J. T. Baker Chemical Co., Phillipsburg, NJ, USA) with
a Teflon-coated pestle, followed by centrifugation at l.OOOX g for 10 min. The
supernatant was collected and centrifuged at 9.000X g for 20 min. The resultant Pa pellet
was resuspended in ice-cold 50mM sodium potassium phosphate (J.T. Baker) buffer (pH
7.4) containing 200mM NaCl (J.T. Baker) and used immediately in binding assays. A
2nM concentration of [35S]TBPS (86 Ci/mmol; New England Nuclear, Boston, MA,
USA) was incubated with 100 jal of tissue homogenate (10% w/v) in the presence or
absence of 5 ^M GABA (Sigma Chem. Co., St. Louis, MO) and 5 (J.1 aliquots of test drug
dissolved in dimethyl sulfoxide (Sigma Chem. Co.) ( assays). At the concentration ( [35S]TBPS binding. All assays were brought to a final volume of 1 ml with 50 mM
sodium potassium phosphate buffer (pH 7.4) containing 200 mM NaCl. Non-specific
binding was defined as binding in the presence of 2p.M TBPS (NEN, Boston, MA) and
accounted for ~30% of the total binding. Assays were terminated after a 90-min steadystate
incubation at 25°C by rapid filtration through glass fiber filters (no. 32; Schleicher
& Schuell, Keene, N.H.). The dissociation kinetics of [35S]TBPS binding were
measured by initiating dissociation by the addition of a saturating concentration of a
known inhibitor of [35S]TBPS binding or a test compound to tissue homogenates preequilibrated
with 2 nM [3SS]TBPS followed by filtration at various time points after the
addition of the known inhibitor or test compound. Allosteric modulators of the known
inhibitor or test compound will modify the rate of dissociation under these conditions
whereas agents acting at common site will not affect the rate. Filter-bound radioactivity
was quantified by liquid scintillation spectrophotometry. The data were evaluated by
nonlinear regression (GraphPad, Inc., San Diego, CA) to obtain ICso (concentration at
which half-maximal inhibition of radioligand occurs) values.
In Vitro Binding Assay 2
[3H]Flunitraiepam binding: assays were carried out under identical conditions, using an
identical tissue preparation, as those used in the [35S]TBPS binding assays with the
exception that 1 |iM GABA was added to all samples instead of 5 uM GABA.
[;!H]Flunitrazepam, 0.2 nM (75 Ci/mmol, New England Nuclear, Boston, MA) was used
to label BZ sites. Non-specific binding is defined as binding in the presence of 1 //M
clonazepam. The data were evaluated by nonlinear regression to obtain ICso and ECjo
values.
In Vitro Binding Assay 3
[3H]Muscimol binding assay: The cortex from male Sprague-Dawley rats (160-200g)
was removed immediately after euthanizing and dissected over ice. The tissue was
homogenized in 15 vol of 0.32M sucrose followed by centrifugation for 10 min at 1000
X g. The supernatant was transferred to a 38 mL polycarbonate tube (Beckrnan
Instruments, Palo Alto CA) and centrifuged at 20,000 X g for 20 min. The membrane
pellet was resuspended in 10 vol of dHjO and centrifuged at 8,000 X g for 20 min. The
resulting pellet was washed with dHbO once and with Na+-free assay buffer (40mM
KH2PC>4, lOOmM KC1, pH 7.4). The pellet was resuspended in 35 mL of Na+-free assay
buffer, incubated at 37°C for thirty minutes and then centrifuged 31,000 X g for twenty
minutes. The final pellet was resuspended in 10 vol of Na+-free assay buffer. Protein
concentration of membrane preparations was ~1 mg/mL by BCA reagent protein assay.
Aliquots of membrane suspension (100 /^L) were incubated in Na+-free assay buffer with
5 nM [3H]muscimol (25 Ci/mmol, New England Nuclear, Boston, MA) and 5 /jL of
dimethylsulfoxide (DMSO) or drug dissolved in DMSO. The final volume of the
incubation medium was 1 mL. Non-specific binding was defined as binding in the
presence of 1 mM GABA. After addition of membranes, tubes were briefly vortexed
and incubated at 4°C in the dark. The incubation was terminated after 60 min by rapid
filtration through glass fiber filters followed by three washes with ice-cold assay buffer.
Filter-bound radioactivity was quantified by LSC after an overnight extraction. The data
were evaluated by nonlinear regression to obtain ICso and ECso values.
Electrophysiological Assay 1.
Pregnant Sprague-Dawley rats, incubating embryos of 17-19 days gestation, were
killed by cervical dislocation. The embryos were removed under aseptic conditions and
the brains quickly excised and placed in Hank's balanced salt solution (HBSS, Gibco) at
ambient room temperature (18-22°C). The hippocampi were dissected out and chopped
into fragments (~ 2mm3) and transferred into an enzyme solution containing (in mM):
NaCl 116, KC1 5.4,NaHC0326,NaH2P04 1, CaCl2 1.5, MgSO4 1, EDTA 0.5, glucose
25, cysteine 1, and papain 20 U/ml (Sigma) and incubated at 37°C, 5% CCh, 100%
relative humidity for 1 hr. Tissue fragments were washed in HBSS containing 1 mg/ml
of bovine serum albumin (BSA) and 1 mg/ml of ovomucoid (both Sigma). Tissue was
transferred into a farther 3-4 ml of this solution and gently triturated into a single cell
suspension using a fire-polished Pasteur pipette. The single cell suspension was layered
on to 5 ml HBSS containing 10 mg/ml of BSA and 10 mg/ml of ovomucoid and
centrifuged at 100 X g for 10 min. The supernatent was discarded and the cells
resuspended in 3-4 ml of glutamine-free minimal essential media (MEM, Gibco)
supplemented with heat-inactivated fetal calf serum (5% v/v Gibco), heat-inactivated
horse serum (5% v/v Gibco), streptomycin and penicillin (50 fig/ml and 5000 i.u./ml,
respectively), glutamine and glucose (final concentrations 2mM and 20mM [Gibco and
BDH] respectively). Approximately 1-2 x 105 cells were plated out on to each 35 mm
(Falcon "Primaria") tissue culture dish which contained ~ 1 ml of the sera-enriched
MEM. The plates were maintained at 37°C, in 5% COi, and 100% relative humidity
until used in electrophysiological studies. Background proliferation of non-neuronal
elements was suppressed with cytosine arabinoside (10 jiM, Sigma) for 48 hr 7 days after
initial dissociation.
Agonist evoked membrane currents were recorded from hippocampal neurons
using the whole cell configuration of the patch-clamp technique. Neurons were voltaged
clamped at -60 mV using a List electronics L/M EPC-7 converter head stage and
amplifier. Cells were perfused with an external (bath) recording solution containing (in
mM): NaCl 140, KC1 2.8, MgCh 2, CaCh 1 and HEPES-NaOH 10 (pH 7.2).
Tetrodotoxin (TTX, 0.3 uM) was included in the recording solution to suppress synaptic
activity. The external solution was delivered (at ~2 ml/min) by a Watson-Marlow flow
pump via non-sterile tubing, which was connected to a plastic cannula (tip dia 1 mm).
The input cannula was mounted on a Prior® micromanipulator and was positioned in
close ( dish via a 19G needle connected by flexible tubing to an aquarium suction pump. The
recording electrode was filled with an internal solution composed of (in mM): CsCl or
KC1 140, MgCl2 2, CaCl2 0.1, EGTA 1.1 (free Ca2+ ~ 10"8 M), HEPES-NaOH 10, and
ATP-Mg2+ 2. The recording electrodes were fabricated from glass hematocrit tubes
(Kimble sodalime tubes 73811) on a Narishige PB7 two stage electrode puller.
Electrodes were coated within 100 p.m of the tip with "Sylgard" (Dow Coming) and fire
polished just before use. Agonists were applied locally to the soma of a voltage-clamped
neuron by pressure ejection (1.4 Kpa, 10-80 msec, 0.1-0.033 Hz) from the tip of a
modified recording pipette using a Picospritzer II device (General Valve Corporation).
The agonist-containing pipette was positioned within 0.1 mm of the cell using a Leitz
micromanipulator. The microscope and micromanipulators were all mounted on a
vibration-free isolation air table (Wentworth) placed inside a Faraday cage. Agonistevoked
whole cell currents were monitored on a storage oscilloscope (Tektronix 2212),
recorded, after digital pulse code modulation (frequency response 14 kHz, Sony PCM
701), and displayed on Multitrace (Electromed) pen chart recorder (frequency response
0.5 kHz). All drugs, other than the agonists, were applied to cells via the superfusion
system. Agonist-evoked whole cell currents were measured at their peak. Responses in
the presence of drugs expressed as the arithmetic mean ± SEM of responses in the
absence (control) or drugs.
Electrophysiology Assay 2
GABAA subunit transfected HEK cells are maintained at 37°C and 5% COa using
Dulbecco's Modified Eagle's Medium with L-glutamine and no sodium pyruvate (Irvine
Scientific #9031, Irvine CA) and supplemented with 10% fetal bovine serum (Irvine
Scientific #3000), 10 U/ml hygromycin B (Calbiochem #400051), and an antibiotic
cocktail consisting of 100 //g/ml streptomycin sulfate, 0.25 //g/ml amphotericin B, 100
units/ml penicillin G (Gibco 15240-096, Gaithersburg MD). Cells are passed by 2 X
wash with phosphate buffered saline (PBS) pH 7.4 and lifted using 1 X trypsin/EDTA
solution in PBS (0.5 mg/ml trypsin, 0.2 mg/ml EDTA, Irvine Scientific #9342) when
confluency reaches ~90%.
GABAA subunit transfected HEK cells are grown to -70% confluency on slide.
Cells are transferred to a bath that is continuously perfused with extracellular saline. The
extracellular medium contained 145 mM NaCI, 3 mM KC1, 1.5 mM CaCh, 1 mM
MgCl2) 5.5 mM d-glucose, and 10 mM HEPES, pH 7.4 at an osmolarity of 320-330
mosM- Recordings are performed at room temperature using the whole cell patch clamp
technique. The patch pipette solution contained 147 mM W-methyl-D-glucamine
hydrochloride, 5 mM CsCl, 5 mM K2ATP, 5 mM HEPES, 1 mM MgCl2, 0.1 mM CaCl2,
and 1.1 mM EGTA, pH 7.2, at an osmolarity of 315 mosM. Pipette-to-bath resistance is
typically 3-5 Mohms. Cells are voltage clamped at -60 mV, and the chloride
equilibrium potential was approximately 0 mV. Drugs are dissolved in extracellular
medium and rapidly applied to the cell by local perfusion. A motor driven multi-channel
switching system exchanged solutions in approximately 20 ms.
In vivo Pharmacology
Vogel conflict
Adult male rats are randomly divided into groups of 6 rats/group. Animals were
deprived of water overnight (24 hr). Food was freely available at time of thirsting.
Thirty minutes after injection (i.p.) of test drug, positive control drug (diazepam,
Img/kg), or vehicle control rats are placed in a square Plexiglas box containing a
stainless steel bottom connected to one side of a drinkometer circuit. At the other side of
the drinkometer circuit a water bottle, placed so the drink tube extends into the Plexiglas
box, is connected. When a subject drinks from the bottle the circuit is closed and an
electric shock is delivered at the tube after seven licks are recorded. The number of licks
in a 3 min session is recorded. Anti-anxiety agents will increase the number of shocks
the animal is willing to endure to acquire water.
Light-dark transition
Male NSA mice (25-30g) are used. The apparatus consists of an open-topped
box divided into small and large area by a partition that has a hole at floor level. The
small compartment is painted black and the large compartment white. The white
compartment was illuminated with light and the black compartment with red light. The
time spent in the light versus dark compartments and the number of transitions between
compartments is recorded during a 5 min test session. Vehicle or test compounds are
administered 30 min prior to the test. Diazepam is administered (i.p.) at 2 mg/kg as the
positive control. Anti-anxiety agents will reduce the time the animals will spend in the
dark compartment and increase the number of transitions between the two compartments.
Carriers
In addition to administering the compound as a raw chemical, the compounds of
the invention may be administered as part of a pharmaceutical preparation containing
suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries,
which facilitate processing of the compounds into preparations, which can be used
pharmaceutically. Preferably, the preparations, particularly those preparations which can
be administered orally and which can be used for the preferred type of administration,
such as tablets, dragees, and capsules, and also preparations which can be administered
rectally, such as suppositories, as well as suitable solutions for administration by
injection or orally, contain from about 0.01 to 99 percent, preferably from about 0.25 to
75 percent of active compound(s), together with the excipient.
Suitable excipients are, in particular, fillers such as saccharides, for example
lactose or sucrose, mannitol or sorbitol, cellulose preparations and/or calcium
phosphates, for example tricalcium phosphate or calcium hydrogen phosphate, as well as
binders such as starch paste, using, for example, maize starch, wheat starch, rice starch,
potato starch, gelatin, tragacanth, methyl cellulose, hydroxypropylmethylcellulose,
sodium carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired, disintegrating
agents may be added such as the above-mentioned starches and also carboxymethylstarch,
cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as
sodium alginate. Auxiliaries are, above all, flow-regulating agents and lubricants, for
example, silica, talc, stearic acid or salts thereof, such as magnesium stearate or calcium
stearate, and/or polyethylene glycol. Dragee cores are provided with suitable coatings
that, if desired, are resistant to gastric juices. For .this purpose, concentrated saccharide
solutions may be used, which may optionally contain gum arabic, talc, polyvinyl
pyrrolidone, polyethylene glycol and/or titanium dioxide, lacquer solutions and suitable
organic solvents or solvent mixtures. In order to produce coatings resistant to gastric
juices, solutions of suitable cellulose preparations such as acetylcellulose phthalate or
hydroxypropymethyl-cellulose phthalate, are used. Dye stuffs or pigments may be added
to the tablets or dragee coatings, for example, for identification or in order to characterize
combinations of active compound doses.
Other pharmaceutical preparations, which can be used orally, include push-fit
capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a
plasticizer such as glycerol or sorbitol. The push-fit capsules can contain the active
compounds in the form of granules, which may be mixed with fillers such as lactose,
binders such as starches, and/or lubricants such as talc or magnesium stearate and,
optionally, stabilizers. In soft capsules, the active compounds are preferably dissolved or
suspended in suitable liquids, such as fatty oils, or liquid paraffin. In addition, stabilizers
may be added.
Possible pharmaceutical preparations, which can be used rectally, include, for
example, suppositories, which consist of a combination of one or more of the active
compounds with a suppository base. Suitable suppository bases are, for example, natural
or synthetic triglycerides, or paraffin hydrocarbons. In addition, it is also possible to use
gelatin rectal capsules, which consist of a combination of the active compounds with a
base. Possible base materials include, for example, liquid triglycerides, polyethylene
glycols, or paraffin hydrocarbons.
Suitable formulations for parenteral administration include aqueous solutions of
the active compounds in water-soluble form, for example, water-soluble salts and
alkaline solutions. In addition, suspensions of the active compounds as appropriate oily
injection suspensions may be administered. Suitable lipophilic solvents or vehicles
include fatty oils, for example, sesame oil, or synthetic fatty acid esters, for example,
ethyl oleate or triglycerides or polyethylene glycol-400 (the compounds are soluble in
PEG-400). Aqueous injection suspensions may contain substances, which increase the
viscosity of the suspension, and include, for example, sodium carboxymethyl cellulose,
sorbitol, and/or dextran. Optionally, the suspension may also contain stabilizers.
Example 1
7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo~l,4-dihydroquinoline-
3-carboxylic acid
To a suspension of 7-chloro-l-ethyl-6-fluoro-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid (Acros; 6.02 g, 22.3 mmol) in 30 mL of l-methyl-2-pyrrolidinone was
added neat 1,2,3,4-tetrahydro-l-aminonaphthalene (20 mL, 20.5 g, 140 mmol) drop-wise
via syringe. The resulting light yellow mixture was placed in an oil bath at 140°C for 17
h. Once at rt, the reaction was added to 120 mL of an aq. 2N HC1 solution and ice. The
solid that formed was isolated by filtration, washed with an aq. 2N HC1 solution (120
mL), water (2 x 100 mL), MeOH (3 x 50 mL) and EtOAc (50 mL). The solid that
remained was then recrystallized from MeOH (1400 mL). The yellow needles that
formed were isolated and washed with methanol (2 x 50 mL). This solid was then
subjected to flash column chromatography. A solution of the solid in 35 mL of 4%
MeOH/CHaCli was added to 16 cm of silica in a 5 cm dia. column. Elution with 1L of
7.5% and 500 mL of 10% MeOH/CH2Cl2 gave 968 mg (11%) of the title compound as a
yellow solid, mp 246-247°C. 'HNMR(400 MHz, DMSO-d6) 8 15.14 (s, 1 H), 8.66 (s, 1
H), 7.80 (s, 1 H), 7.63 (s, 1H), 7.32 (d, 1 H, J = 7.7 Hz), 7.26-7.16 (m, 3 H), 4.90 (s, 2
H), 4.33 (q, 2 H, J = 7.2 Hz), 2.92-2.80 (m, 2 H), 112-2.00 (m, 2 H), 1.92-1.86 (m, 2 H),
1.60 (t, 3 H, J = 7.2 Hz). MS (M + Na)+ 419. Anal Calcd. for C22H2iClN2O3 + 0.25
HC1: C, 65.08; H, 5.28; Cl, 10.92; N, 6.90. Found: C, 65.09; H, 5.33; Cl, 10.85; N, 6.81.
The following compounds were prepared by using the method described above:
(R)-7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid;
(S)-7-Chloro-1 -ethy l-6-( 1,2,3,4-tetrahydronaphthyl-1 -amino)-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro-l -ethyl-6-(6-methoxy-l ,2,3,4-tetrahydronaphthyl-l -amino)-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro-l-ethyl-6-(l-aminoindanyl)-4-oxo-l,4-dihydroquinoline-3-carboxylic
acid;
7-Chloro-1 -ethyl-6-(5 -methyl-1 -aminoindanyl)~4-oxo-1,4-dihydroquinoline-3 -
carboxylic acid;
21
7-Chloro-l-ethyl-6-(2-aminoindanyl)-4-oxo-l,4-dihydroquinoline-3-carboxylic
acid;
7-Chloro-l-ethyl-6-(benzylamino)-4-oxo-l,4-di"hydroquinoline-3-carboxylic
acid;
7-Chloro-1 -ethyl-6-(2-phenethylamino)-4-oxo-1,4-dihydroquinoline-3-carboxylic
acid. The reaction was performed as in Example 1 except that the crude reaction mixture
was diluted with EtOAc giving the desired compound as a white solid. *H NMR (400
MHz, DMSO-d6) 8 15.15 (br s, 1H), 8.64 (s, 1H), 7.63 (s, 1H), 7.58 (s, 1H), 7.37-7.33
(m, 2H), 7.29-7.24 (m, 3H), 4.68 (t, 1H, J = 5.4 Hz), 4.31 (q, 2H, J = 7.3 Hz), 3.59 (q,
2H, J = 6.4 Hz), 3.03 (t, 2H, J = 6.9 Hz), 1.58 (t, 3H, J = 7.3 Hz);
7-Chloro-l-methyl-6-(l,2)3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid, *H NMR (400 MHz, CDCh) 8 15.10 (s, 1H), 8.62
(s, 1H), 7.77 (s, 1H), 7.62 (s, 1H), 7.33 (d, 1H, J = 7.0 Hz), 7.25-7.17 (m, 3H), 4.91 (br s,
2H), 3.99 (s, 3H), 2.86 (m, 2 H), 2.11-2.01 (m, 2 H), 1.91-1.87 (m, 2H);
7-Chloro-l-ethyl-6-[4-methoxy(phenethylamino)]-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-cyclopropyl-6-[4-methoxy(phenethylamino)]-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro-1 -ethyl-6-[3-methoxy(phenethylamino)]-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-[2-methoxy(phenethylamino)]-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-[4-bromo(phenethylamino)]-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-[4-chloro(phenethylamino)]-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-[4-fluoro(phenethy]amino)]-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-(3-phenylpropylamino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-ChIoro-l-ethyl-6-(4-phenylbutylamino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
22
7-Chloro-l-ethyl-6-(4-phenylbutyl-2-amino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-(2-phenylcyclopropylamino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-(2-phenylcyclopropylamino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l -ethyl-6-( 1 -naphthylethyl-l-amino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-l-ethyl-6-(l-naphthylmethylamino)-4-oxo-lJ4-dihydroquinoline-3-
carboxylic acid and
7-Chloro-l-ethyl-6-(2-phenoxyethylamino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid.
Example 2
7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-
4-oxo-l,4-dihydroquinoline-3-(n-propyl)carboxamide
To a solution of 7-chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxol,
4-dihydroquinoline-3-carboxylic acid (37 mg, 0.093 mmol) in 5 mL of CHCh at -10
°C was added neat Et3N (30 (iL, 22 mg, 0.22 mmol) and benzyl chloroformate (17 (iL, 20
mg, 0.117 mmol). After stirring cold for 45 m, neat propylamine (10 jaL, 7.2 mg, 0.122
mmol) was added via syringe to the reaction at -20 °C. The reaction was then allowed to
warm to rt over 2 h and added to 7 mL each of a 10% aq. I^COj solution and CHCh.
The organic layer was separated and washed with water (2x10 mL), dried (IS^SO-O,
filtered and concentrated to dryness. The residue was taken up in CH2Ch and added to
10 cm of flash silica gel in a 2 cm dia. column. Elution with 100% CKfeCh (100 mL) and
2.5% MeOH/CH2Cl2 (200 mL) gave 37 mg (91%) of the title compound as a yellow
solid.
The following compounds were prepared by using the general method given in
Example 2:
7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-(2-phenethyl)carboxamide;
7-Chloro-l-ethyl-6-(l ,2,3,4-tetrahydronaphthyl-l -amino)-4-oxo-l ,4-
23
dihydroquinoline-3-(2-dimethylaminoethyl)carboxamideand
7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-(pyridylmethylamino)carboxamide.
Example 3
7-Chloro-l-(2-phenethyl)-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid
a. Ethyl 2-(2,4-dichloro-5-fluorobenzoyl)-3-(dimethylamino)acryIate. A mixture of
ethyl 3,3-dimethylaminoacrylate (3.10 g, 21.6 mmol) and N,N-diisopropylethylamine
(8.0 mL, 5.94 g, 45.9 mmol) was stirred at rt and a solution of 2,4-dichloro-5-
fluorobenzoyl chloride (4.92 g, 21.6 mmol) was added drop-wise via addition funnel
over 20 m. The cloudy yellow solution that formed was placed in an oil bath at 85-90
°C. After 3 h, the mixture that formed was filtered and the solid was washed with
benzene. The dark filtrate was concentrated and the residue was triturated with hexanes
(50 mL). The solid that didn't dissolve was collected and washed with hexanes (20 mL),
The resulting solid was partitioned between water and EtOAc. The EtOAc layer was
washed with water (3 x 25 mL), brine, dried (NazSO.}), filtered and concentrated to 5.0 g
(69%) of the desired compound.
b. Ethyl 2-(2,4-dichloro-5-fluorobenzoyl)-3-(2-phenethylamino)acrylate. A
suspension of ethyl 2-(2,4-dichloro-5-fluorobenzoyl)-3-(dimethylamino)acrylate (1.014
g, 3.03 mmol) in 10 mL of EtOH was treated with neat phenethylamine (0.4 mL, 386
mg, 3.19 mmol) added drop-wise via syringe. After stirring at rt for 75 m, the mixture
that formed was filtered and the solid was washed with EtOH leaving 620 mg (50%) of
the acrylate as a white solid.
c. Ethyl 7-chIoro-6-fluoro-l-(2-phenethyl)-4-oxo-l,4-dihydroquinoline-3-
carboxylate. To a solution of ethyl 2-(2,4-dichloro-5-fluorobenzoyl)-3-(2-
phenethylamino)acrylate (656 mg, 1.60 mmol) in 1.5 mL of DMF was added solid
K^COj (227 mg, 1.64 mmol). The resulting mixture was placed in an oil bath at 130 °C
for 16 h. Once at rt, the reaction was added to water. The gummy solid that formed was
partitioned between water and EtOAc. The aq. layer was extracted with EtOAc (2x10
mL). The pooled organic layers were washed with water (2x15 mL), brine and dried
24
). The solvent was removed in vacuo, giving 572 mg (96%) of the desired
quinolone.
d. 7-Chloro-6-fluoro-l-(2-phenethyl)-4-oxo-l,4-dihydroquinoline-3-carboxylic acid.
A solution of ethyl 7-chloro-6-fluoro-l-(2-phenethyl)-4-oxo-l,4-dihydroquinoline-3-
carboxylate (516 mg, 1.38 mmol) in 5.7 mL of an aq. 6 N HC1 solution was placed in an
oil bath at 113 °C for 3h 40 m. Once at rt, the mixture was filtered and washed with
water (2x10 mL) to give 466 mg (98%) of the acid as a solid.
e. 7-Chloro-l-(2-phenethyl)-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxyIic acid. Using the procedure described in Example 1, the
title compound was isolated in 6% yield.
By using the method in Example 3, the following compounds were prepared:
7-Chloro-l-(benzyl)-6-(4-phenylbutyl-2-amino)-4-oxo-l,4-dihydroquinoline-3-
carboxylic acid.
25
Example 4
Modulation off5S]TBPS binding in rat cortex by 7-chloro-l-ethyl-6-(l,2,3,4-
tetrahydronaphthyl-l-amino)-4-oxo-l,4-dihydroquinoline-3-carboxylic acid
Theability of T-chloro-l-ethyl-e-Cl^.S^-tetrahydronaphthyl-l-amino^-oxol,
4-dihydroquinoline-3-carboxylic acid to inhibit the binding of [35S]TBPS was
determined according to the previously described method. The following compounds in
Tables 1 and 2 were also tested for their ability to inhibit or enhance [ 5S]TBPS binding
to rat cortex.
(Table Removed)






WE CLAIM:
1. A substituted quinolone having the Formula I
(Formula Removed)
or a pharmaceutically acceptable salt, amide, ester or solvate thereof,
wherein:
R1 is selected from the group consisting of hydrogen; an optionally
substituted C1-20 alkyl, and C7-30 aralkyl;
each R2 is selected from the group consisting of hydrogen and optionally
substituted C1-20 alkyl;
each R3 is selected from the group consisting of hydrogen, optionally
substituted C1-20 alkyl; a group OR11 and NR12R13;
R5, R7 and R8 are independently selected from the group consisting of
hydrogen, an optionally substituted C1-20 alkyl, and halogen;
R9 and R1o are independently selected from the group consisting of
hydrogen, optionally substituted C1-20 alkyl, C7-30 aralkyl, C3-8 cycloalkyl and
C6-22 cycloaralkyl; with the proviso that R9 and R1o are not both hydrogen at
the same time;
R11 is selected from the group consisting of hydrogen, an alkali metal, a
negative charge and optionally substituted C1-20 alkyl;
R12 and R13 are independently selected from the group consisting of
hydrogen, optionally substituted C1-20 alkyl, C7-30 aralkyl, C6-14 aryl, C3-8
cycloalkyl and C6-22 cycloaralkyl; or R12 and R13 are taken together with the
nitrogen atom to which they are attached to form a heterocyclic ring.
2. A compound as claimed in claim 1, having the Formula II:
(Formula Removed)


or a pharmaceutically acceptable salt, amide, ester or solvate thereof.
3. A compound as claimed in claim 1, having the Formula III:
(Formula Removed)
or a pharmaceutically acceptable salt, amide, ester or solvate thereof, wherein n is an integer 0, 1, 2, 3 or 4.
4. The compound as claimed in claim 3, wherein n is 2.
5. The compound as claimed in claim 3, wherein R1 is alkyl, R2, Rs and R8 are hydrogen and R7 is halogen.
6. A compound as claimed in claim 1, wherein said compound is: 7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-dihydroquinoline-3-carboxylic acid;
(R)-7-Chloro-1 -ethyl-6-( 1,2,3,4-tetrahydronaphthyl-1 -amino)-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
(S)-7-Chloro-l-ethyl-6-(l,2,3,4-tetrahydronaphthyl-l-amino)-4-oxo-l,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro-1 -ethyl-6-( 1 -aminoindanyl)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-1 -ethyl-6-(2-aminoindanyl)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-1 -ethyl-6-(benzylamino)-4-oxo-1,4-dihydroquinoline-3-carboxylic
acid;

7-Chloro-1 -ethyl-6-(phenethyl-2-amino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro- l-ethyl-6-[4-methoxy(phenethylamino)]-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro- l-ethyl-6-[3-methoxy(phenethylamino)]-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro-1 -ethyl-6-[2-methoxy(phenethylamino)]-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
7-Chloro-1 -ethyl-6-[4-bromo(phenethylamino)]-4-oxo-1,4-dihydroquinoline-
3-carboxylic acid;
7-Chloro-1 -ethyl-6-[4-chloro(phenethylamino)]-4-oxo-1,4-dihydroquinoline-
3-carboxylic acid;
7-Chloro- l-ethyl-6-(3-phenylpropylamino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-1 -ethyl-6-(4-phenylbutylamino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-1 -ethyl-6-(4-phenylburyl-2-amino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-1 -ethyl-6-(2-phenylpropylamino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid;
7-Chloro-1 -ethyl-6-(2-phenoxyethylamino)-4-oxo-1,4-dihydroquinoline-3-
carboxylic acid, or
7-Chloro-1 -methyl-6-( 1,2,3,4-tetrahydronaphthyl-1 -amino)-4-oxo-1,4-
dihydroquinoline-3-carboxylic acid;
or a pharmaceutically acceptable salt, amide, ester or solvate thereof.
7. A pharmaceutical composition as and when prepared by using a compound of Formula I as claimed in any of the preceding claims:

(Formula Removed)
or a pharmaceutically acceptable salt, amide, ester or solvate thereof, and a pharmaceutically acceptable carrier selected from the group consisting of excipients and auxiliaries.

Documents:

3419-DELNP-2004-Abstract-(13-10-2008).pdf

3419-DELNP-2004-Abstract-(26-09-2008).pdf

3419-delnp-2004-abstract.pdf

3419-DELNP-2004-Claims-(13-10-2008).pdf

3419-DELNP-2004-Claims-(26-09-2008).pdf

3419-delnp-2004-claims.pdf

3419-delnp-2004-complete specification(granted).pdf

3419-DELNP-2004-Correspondence-Others-(26-09-2008).pdf

3419-delnp-2004-correspondence-others.pdf

3419-delnp-2004-description (complete)-(13-10-2008).pdf

3419-DELNP-2004-Description (Complete)-(26-09-2008).pdf

3419-delnp-2004-description (complete).pdf

3419-DELNP-2004-Drawings-(26-09-2008).pdf

3419-delnp-2004-drawings.pdf

3419-DELNP-2004-Form-1-(13-10-2008).pdf

3419-DELNP-2004-Form-1-(26-09-2008).pdf

3419-delnp-2004-form-1.pdf

3419-delnp-2004-form-13.pdf

3419-delnp-2004-form-18.pdf

3419-DELNP-2004-Form-2-(13-10-2008).pdf

3419-DELNP-2004-Form-2-(26-09-2008).pdf

3419-delnp-2004-form-2.pdf

3419-delnp-2004-form-3.pdf

3419-delnp-2004-form-5.pdf

3419-DELNP-2004-GPA-(26-09-2008).pdf

3419-delnp-2004-gpa.pdf

3419-delnp-2004-pct-210.pdf

3419-delnp-2004-pct-304.pdf

3419-delnp-2004-pct-409.pdf

3419-delnp-2004-pct-416.pdf


Patent Number 227725
Indian Patent Application Number 3419/DELNP/2004
PG Journal Number 05/2009
Publication Date 30-Jan-2009
Grant Date 19-Jan-2009
Date of Filing 03-Nov-2004
Name of Patentee THE REGENTS OF THE UNIVERSITY OF CALIFORNIA
Applicant Address 1111 FRANKLIN STREET, 12th FLOOR, OAKLAND, CA 94607, UNITED STATES OF AMERICA.
Inventors:
# Inventor's Name Inventor's Address
1 DERK J. HOGENKAMP 4510 PARK DRIVE, CARLSBAD, CALIFORNIA 92008, USA.
2 KELVIN W. GEE 47 UREY COURT, IRVINE, CALIFORNIA 92612,USA.
3 TIMOTHY B.C. JOHNSTONE 2289 SANTA ANA AVENUE, #B, COSTA MESA, CALIFORNIA 92627, USA.
PCT International Classification Number A61K 31/47
PCT International Application Number PCT/US2003/014948
PCT International Filing date 2003-05-12
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 60/380,641 2002-05-14 U.S.A.