Note: Descriptions are shown in the official language in which they were submitted.
WO 2012/018635 CA 02806664 2013-01-25 PCT/US2011/045390
ARYLSULFONAMIDE DERIVATIVES, COMPOSITIONS, AND METHODS OF USE
Cross-Reference to Related Patent Applications
[0001] This International Patent Application claims priority to U.S.
Provisional Patent Application
No. 61/367,709, filed July 26, 2010, the disclosure of which is herein
incorporated by reference.
Field of the Invention
[0002] The present invention relates to arylsulfonamide derivatives,
positional isomers, and
prodrugs thereof, compositions comprising the same and methods of making and
using the same.
The present invention also relates to pharmaceutical compositions comprising
these compounds and
methods for using these compounds. Compounds described herein are particularly
useful for the
treatment and/or prophylaxis of diseases, disorders, and conditions that
involve the Na+KFC1- co-
transporters (NKCC1 or NKCC2 or combinations thereof) including but not
limited to addictive
disorders, anxiety disorders, ascites, attention deficit hyperactivity
disorder (ADHD), bipolar
disorder, cancer, depression, edema, endothelial corneal dystrophy, epilepsy,
glaucoma,
inflammatory pain, ischemia, migraine, neuropathic pain, nociceptive pain,
ocular diseases, pain,
postherpetic neuralgia, and schizophrenia. Compounds described herein are also
particularly useful
for the treatment and/or prophylaxis of diseases, disorders, and conditions
that involve GABAA
receptors including but not limited to Alzheimer's Disease, addictive
disorders, anxiety disorders,
attention deficit hyperactivity disorder (ADHD), autism spectrum disorders
(autism), bipolar
disorder, cognitive function (e.g., cognitive impairment, cognitive
dysfunction), depression,
epilepsy, Huntington's Disease, inflammatory pain, insomnia, migraine,
migraine with aura,
migraine without aura, neuropathic pain, nociceptive pain, pain, Parkinson's
disease, periodic limb
movement disorder (PLMD) (nocturnal myoclonus), personality disorders,
postherpetic neuralgia,
psychosis, restless legs syndrome (RLS), schizophrenia, seizure disorders,
spasticity, tinnitus, and
withdrawal syndromes.
Background of the Invention
Na4(+0- Co-Transporters
[0003] In absorptive and secretory epithelia, transcellular ion transport
depends on specific plasma
membrane proteins for mediating ion entry and exit from cells. In basolateral
membrane of almost
all epithelia (with exception of choroidal plexus), sodium exit and potassium
entrance occur through
NeKtATPase, generating electrochemical gradients that constitute a driving
force for Na+ influx
and K4 efflux. Transport of these ions following their gradients can be
accomplished by specific ion
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WO 2012/018635 CA 02806664 2013-01-25 PCT/US2011/045390
channels, allowing membrane passage of ions alone or by transporters in which
Na + or K+ transport
is accompanied by other ions or solutes by means of several different solute
transporters. These
membrane proteins are known as secondary transporters because ion
translocation is not dependent
on ATP hydrolysis but rather on gradients generated by primary transporters. A
secondary transport
mechanism for transcellular ion transport in epithelial cells involves cations
(Nat or K+) are coupled
with chloride (Cr), with a stoichiometry of 1:1, and, therefore, the ion
translocation produces no
change in transmembrane potential. For this reason, these transporters are
known as electroneutral
cation-chloride coupled cotransporters. In addition to being heavily
implicated in ion absorptive and
secretory mechanisms, electroneutral cation-chloride coupled cotransporters
play a key role in
maintenance and regulation of cell volume in both epithelial and nonepithelial
cells. Because Na+
influx and K+ efflux by electroneutral cotransporters are rapidly corrected by
NeKtATPases, the
net effect of its activity is Cl- movement inside or outside cells. The change
in intracellular chloride
concentration is known to be accompanied by changes in cell volume. Finally, a
variety of new
physiological roles for electroneutral cotransporters are emerging (e.g.,
regulation of intraneuronal
Cl- concentration and thus modulation of neurotransmission). Gamba (2005)
Physiol. Rev. 85: 423-
493.
[0004] Four groups of electroneutral cotransporter systems (also known as
"symporters") have
been functionally identified based on cation(s) coupled with chloride,
stoichiometry of transport
process, and sensitivity to inhibitors. These systems include: (1) the
benzothiadiazine (or thiazide)-
sensitive Na.+0- cotransporter; (2) the sulfamoylbenzoic (or bumetanide)
sensitive Na+K+20-
cotransporters; (3) the sulfamoylbenzoic (or bumetanide) sensitive NaCl -
cotransporters; and (4) the
dihydroindenyloxy-alkanoic acid (DIOA)-sensitive ic+cr cotransporter. Some
overlap exists in
sensitivity to inhibitors in the last two groups because Ner2C1- and KCl
cotransporters can be
inhibited by high concentration of DIOA or loop diuretics, respectively;
however, affinity for
inhibitor and the cation coupled with chloride clearly differentiate between
both groups of
transporters. Gamba (2005) Physiol. Rev. 85: 423-493. Loop diuretics (e.g.,
bumetanide,
furosemide, piretanide, azosemide, and torsemide) are antagonists of the
Na+K+Cl- cotransporter
(e.g., NKCC2) in the thick ascending limb of the loop of Henle and act to
inhibit sodium and
chloride reabsorption by competing for the Cl- binding site. See also Russell
(2000) Physioglocal
Reviews 80(1): 211-275.
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[0005] Major advances have been made in the past decade in molecular
identification and
characterization of solute carriers. As of 2005, the Human Genome Organization
(HUGO)
Nomenclature Committee Database recognizes 43 solute carries (SLC) families,
which include a
total of 298 transporter genes encoding for uniporters (passive transporters),
cotransporters (coupled
transporters), antiporters (exchangers), vesicular transporters, and
mitochondria' transporters. This
amount of solute carrier genes represents ¨1% of the total pool of genes that
have been calculated to
compose human genome. Gamba (2005) Physiol. Rev. 85: 423-493.
[0006] One isoform of the Na.+K+Cl- cotransporter (NKCC) NKCC1 is widely
distributed
throughout the body. NKCC1 transports sodium, potassium, and chloride into the
cell. NKCC1 is
also found throughout the nervous system where it is expressed on astrocytes,
oligodendrocytes, and
Schwann cells. Lenart, et al. (2004) The Journal of Neuroscience 24(43): 9585-
9597. Another
isoform, NKCC2, is found primarily in the kidney, where it serves to extract
sodium, potassium, and
chloride from the urine. Haas (1994) Am J Physiol Cell Physiol 267: C869¨C885.
[0007] The mediators of transcellular Cr cotransport (Na-Cl cotransporter,
NKCC1, NKCC2,
KCC1, and KCC2) are all related members of the SLC12A family of cation/Cr
cotransporters; each
takes advantage of inward Na + or outward K gradients to move Cr into or out
of cells, respectively.
The importance of this family of transporters are underscored by their use as
pharmacologic targets
(thiazide diuretics act at NKCC, and loop diuretics act at NKCC2), and that
their mutation results in
diverse diseases. For example, disruption of NKCC1 in mice leads to hearing
loss, altered pain
perception, neuronal excitability, and altered blood pressure. Kahle, et al.
(2004) Proc. Natl. Acad.
Sci. USA 102(46): 16783-16788.
[0008] The regulation of Cl- transport into and out of cells also plays a
critical role in the
maintenance of intracellular volume and the excitability of GABA responsive
neurons regulated by
at least two ion cotransporters: Cl- influx is mediated by the NKCC1 which
mediates the Cr influx
and KCC I or KCC2 which mediates the Cr efflux. Kahle, et al. (2004) Proc.
Natl. Acad. Sci. USA
102(46): 16783-16788. The maintenance of intra- and extracellular electrolyte
homeostasis are
required for a wide range of essential physiologic processes, including
general functions (e.g.,
maintenance of proper cell volume), specialized cell functions (e.g., control
of neuronal excitability),
and global functions (e.g., regulation of blood pressure). This homeostasis is
achieved via the
regulated movement of Nat, K+, and Cl- across cell membranes by ion channels,
cotransporters,
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WO 2012/018635 CA 02806664 2013-01-25 PCT/US2011/045390
exchangers, and pumps that execute transmembrane electrolyte flux. Kahle, et
al. (2004) Proc. Natl.
Acad. Sci. USA 102(46): 16783-16788.
[0009] The predominant mechanism by which intracellular volume is maintained
in cells in
response to changes in extracellular tonicity is the raising or lowering of
intracellular CF
concentration ([01i), thereby minimizing transmembrane water flux. [C11; is
modulated by altering
the balance between CF entry and exit. The major mediator of CF entry is NKCC1
and Cr exit is
largely mediated by KCC1. These cotransporters are both regulated by
extracellular tonicity:
hypertonicity activates NKCC1 and inhibits KCC1, whereas hypotonicity has the
opposite effect.
Kahle, et al. (2004) Proc. Natl. Acad. Sci. USA 102(46): 16783-16788.
[0010] An analogous system plays a key role in the control of neuronal
excitability where,
variation in [CF] in neurons is determined by mechanisms highly similar to
those governing cell
volume. CF influx largely occurs via NKCC1, whereas CF efflux is mediated via
the neuronal-
specific K-Cl cotransporter KCC2. Kahle, et al. (2004) Proc. Natl. Acad. Sci.
USA 102(46): 16783-
16788.
GABA Receptors
[0011] Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter
in the central
nervous system (CNS) where approximately 30% of all synapses use GABA as a
transmitter. Thus,
GABA receptors are crucial for proper cognitive function and balancing of
excitatory and inhibitory
signals in the brain. There are three classes of GABA receptors: GABAA (ligand-
gated ion channel),
GABAB (G protein-coupled receptor), and GABAc(ligand-gated ion channel).
Chloride flux into
the cell results from the activation of GABAA receptors by the binding of GABA
molecules,
hyperpolarizing the resting membrane potential, and decreasing the chances of
the post-synaptic
neuron propagating an action potential.
[0012] GABAA receptors are pentameric and approximately 19 GABA receptor
subunits have been
cloned from mammals (6 a, 3 13, 3 7, 1 5, 1 a, 10, 1 IT, and 3 p subunits).
The heterogeneity of
GABA subunits are further increased by alternate splicing (e.g., 72 short and
y2 long are the two
major splice variants of the 72). In general, a functional GABAA receptor
requires 2 a subunits, 213
subunits and a third "regulatory" subunit (usually 7 or 5). WO 2009/100040.
The specific subunit
combination determines the pharmacological and ligand binding properties of
the GABAA receptor.
The most abundant subunit combination found in the CNS are a10272. This
subtype represents
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WO 2012/018635 CA 02806664 2013-01-25 PCT/US2011/045390
approximately 40% of GABAA receptors in the brain and it is expressed
throughout the CNS and is
located on post-synaptic cells. WO 2007/002359.
[0013] The importance of [C1]1 regulation has been recognized with the
discovery that GABA
neurotransmission is not uniformly inhibitory (e.g., it is predominantly
excitatory in the neonatal
period.) If [CF]1 is below its equilibrium potential, cr enters the cell,
resulting in hypetpolarization
and inhibition. If [C1-]1 is above its equilibrium potential, GABA induces cr
efflux, depolarization,
and neuronal excitation. Similarly, neurons of the suprachiasmatic nucleus
show circadian variation
in their response to GABA, demonstrating the ability to dynamically regulate
[CF]1. Finally, GABA
neurotransmission in the peripheral nervous system is predominantly
excitatory.
[0014] GABAA receptors are the targets of a wide range of therapeutic and
clinically relevant
compounds including benzodiazepines, barbiturates, neurosteroids, ethanol,
certain intravenous
anesthetics, and subtype specific modulators (e.g., Zolpidem.) These compounds
serve as
anxiolytics, sedative/hypnotics, anti-epileptic drugs (AED), and memory
enhancers. Many of these
therapeutics show efficacy but cause side effects due to unwanted effects at
oci and/or oc2 GABAA
variants or due to low therapeutic index. For example, benzodiazepines such as
diazepam
(VALIUM) are excellent anxiolytics but cause unwanted sedative effects. WO
2007/002359.
[0015] At a cellular level, GABAA receptors are expressed both pre-synaptic,
post-synaptic, and
extra-synaptic sites (pre-synaptic and extrasynaptic being defined herein as
parasynaptic to
distinguish from post-synaptic) where they respond to large changes in GABA
concentration caused
by release of the neurotransmitter into the synaptic space, and extra-
synaptically where the receptors
respond to lower concentrations of GABA that "leak" from synaptic junctions.
The post-synaptic
receptors respond to acute changes in neuronal firing, pre-synaptic receptors
are responsible for
inhibition of GABA release in the setting of high GABA levels, whereas the
extrasynaptic receptors
are responsible for maintaining overall tone of neuronal networks. WO
2009/100040. Tonic
inhibition is generated by the persistent activation of extrasynapatic
(perisynaptic) GABAA receptors
and regulates the excitability of individual neurons and neural networks. Jia,
et al. (2008) Ti
Journal of Pharmacology and Experimental Therapeutics 326(2): 475-482.
[0016] Presynaptic GABAA receptors situated at extrasynaptic sites may
comprise a4135 and a6135
isoforms. The extrasynaptic a4f38 and a6[38 GABAA receptor isoforms show
marked sensitivity to
GABA, alcohol, and anesthetics, suggesting that receptors may present a
critical site for regulating
synaptic function in the developing brain in both physiological and
pathological situations. Xiao, et
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WO 2012/018635 CA 02806664 2013-01-25 PCT/US2011/045390
at. (2007) J Physiol. 580(Pt.3):731-43. For example, temporal lobe epilepsy
(TLE), Parkinson's
disease (PD) and Huntington's disease (HD) are neurodegenerative disorders
that involve
disruptions in GABA signaling. TLE seizures reflect excess excitation, which
may result from local
inhibitory circuit dysfunction. PD devastates the input to striatal GABAergic
neurons and HD
destroys striatal GABAergic neurons. Directing GABA synthesis, degradation,
release, transport or
receptors may be useful in controlling GABA signaling in specific brain areas
should benefit each of
these diseases. Thus, new drugs targeting GABA synthesis, release, and binding
may be useful for
improved therapeutic treatments for epilepsy and both Parkinson's and
Huntington's disease.
Kleppner and Tobin (2001) Expert Opin Ther Targets. 5(2):219-39. See also
Shumate, et al. (1998)
Epilepsy Research (32): 114-128; Fritschy (2008) Frontiers in Molecular
Neuroscience 1(5): 1-5;
Roberts, et at. (2006) The Journal of Biological Chemistry 281(40): 29431-
29435; and Roberts, et
al. PNAS 102(33): 11894-11899.
Addictive Disorders
[0017] Addictive and/or compulsive disorders, such as eating disorders
(including obesity),
addiction/physical dependence to stimulants, narcotics (e.g., cocaine, heroin)
sedatives/hypnotics,
and opioids including alcoholism and smoking are major public health problems
that impact society
on multiple levels. It has been estimated that substance abuse alone costs the
United States more
than $484 billion per year.
[0018] The alcohol-sensitive a4135 GABAA receptor has also been postulated to
be involved in
alcohol addiction (alcoholism). For example, reduced expression of GABAA
receptors comprising
an a4 subunit in the nucleus accumbens (NAc) decreased the free consumption
and preference for
alcohol in rats. Further, the nucleus accumbens contributes to the rewarding
and reinforcing effects
of drugs including alcohol suggesting that the GABAA receptor, specifically
the a4135 isoform, in the
NAc is an important mediator of alcohol self-administration. Rewal, et at.
(2009) The Journal of
Neuroscience 29(2): 543-549. Although most GABAA receptor subunit combinations
can be
activated by high (anesthetic) alcohol concentrations, so far only very
specific GABAA receptor
subunit combinations (containing the 5 as well as the f33 subunit) exhibit
dose-dependencies that
mirror blood alcohol levels associated with mild to moderate intoxication in
humans. These 5-
subunit containing GABAA receptors containing the 5 subunit are located either
outside or in the
perimeter of synapses, but not in the sub-synaptic membrane. WO 2007/002359.
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[0019] Current strategies for the treatment of additive disorders include
psychological counseling
and support, use of therapeutic agents, or a combination of both. A variety of
agents known to affect
the central nervous system have been used in various contexts to treat a
number of indications
related directly or indirectly to addictive behaviors but a great need remains
for improved addictive
disorder therapeutics. GABAA specific agents may be effective therapeutics for
addictive behaviors.
Alzheimer's Disease
[0020] Alzheimer's disease (AD) is an age-related, non-reversible brain
disorder that develops
over a period of years and is the most common cause of dementia among people
age 65 and older.
Initially, people experience memory loss and confusion, which may be mistaken
for the kinds of
memory changes associated with normal aging. However, the symptoms of AD
gradually lead to
behavior and personality changes, a decline in cognitive abilities such as
decision-making and
language skills, and problems recognizing family and friends. AD ultimately
leads to a severe loss
of mental function. NINDS Alzheimer's Disease Information Page (2009).
[0021] AD results in neuron death in the brain. As neurons die throughout the
brain, the affected
regions begin to atrophy. By the final stage of AD, damage is widespread and
brain tissue has
shrunk significantly. Two major hallmarks associated with the AD disease
processes in the brain are
amyloid plaques and neurofibrillary tangles. Amyloid plaques comprise
fragments of p-amyloid
peptide mixed with a collection of additional proteins, and remnants of
neurons. Neurofibrillary
tangles (NFTs) are found inside neurons and comprise tau protein. NINDS
Alzheimer's Disease
Information Page (2009).
[0022] Currently there are no medicines that can slow the progression of AD.
However, four
FDA-approved medications are used to treat AD symptoms. Donepezil (Aricept),
rivastigmine
(Exelon), galantamine (Reminyl), and memantine (Namenda) are prescribed to
treat AD symptoms.
NINDS Alzheimer's Disease Information Page (2009). Further, treatment of an AD
transgenic
mouse model with picrotoxin, a GABAA antagonist, showed improved cognitive
functions in the
mice. Yoshiike, et al. (August 21, 2008) PLoS One. 3(8):e3029. Additionally,
the expression of
NKCCI has been found to be elevated in AD patients. Johanson, et al. (2004)
Cerebrospinal Fluid
Research 1:3. Unfortunately these medications will not stop or reverse AD, and
they appear to help
individuals for only a few months to a few years. Therefore novel therapies
based on the regulation
of GABAA receptor activity may relieve the symptoms of AD.
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Anxiety Disorders
[0023] Anxiety disorders are classified into several subtypes: anxiety, acute
anxiety, panic
disorder, social anxiety disorder, obsessive compulsive disorder (OCD), panic
disorder, panic
symptoms, post-traumatic stress disorder (PTSD), generalized anxiety disorder,
and specific phobia.
American Psychiatric Association. Diagnostic and Statistical Manual of Mental
Disorders, 4th
edition (1994).
[0024] As a group, the anxiety disorders have the highest prevalence in the
U.S. of all psychiatric
disorders and afflict 15.7 million people in the United States each year, and
30 million people in the
United States at some point in their lives. Lepine (2002) J. Clin. Psychiatry.
63: Suppl 14:4-8;
Kessler, et al. (1994) Arch Gen Psychiatry 51:8-19.
[0025] Several animal models are recognized in the art as being predictive of
anxiolytic activity.
These include the fear-potentiated startle model, described by Davis in
Psychopharmacology 62:1;
1979, Behav. Neurosci. 100:814;1986 and TiPS, January 1992 Vol. 13,35-41, the
elevated plus
model described by Lister in Psychopharmacol. 92:180-185; 1987, and the well-
known punished-
responding (conflict) model, described, in "Psychopharmacology of Anxiolytics
and
Antidepressants", edited by S. E. File, pages 131-153, Raven Press, New York,
1991.
[0026] Anxiety disorders are generally treated with drugs and psychotherapy.
The most commonly
prescribed drugs for all anxiety types are benzodiazepines and selective
serotonin reuptake inhibitors
(SSRI). However, while these drugs show efficacy, both benzodiazepines and
SSRIs show adverse
effects during treatment. Denys and de Geus (August 2005) Curr Psychiatry Rep.
7(4): 252-7.
Further, numerous side effects are associated with long-term use of SSRIs,
such as sexual
dysfunction and weight gain. Hirschfeld (2003) J. Clin. Psychiatry. 64: Suppl
18: 20-4.
Additionally, existing drugs targeting postsynaptic type GABAA receptors
produce undesirable
results because they indiscriminately target most of the GABAA receptors in
the brain.
WO 2007/136838. In view of nonresponders and deleterious side effects, a great
need exists for
improved anxiety therapeutics.
Aseites
[0027] Ascites are excess fluid in the space between the tissues lining the
abdomen and abdominal
organs (the peritoneal cavity) typically caused by liver disease. Disorders
that may be associated
with ascites include: cirrhosis, hepatitis, portal vein thrombosis,
constrictive pericarditis, congestive
heart failure, liver cancer, ovarian cancer, protein-losing enteropathy,
nephrotic syndrome, and
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pancreatitis. Some agents are available for the treatment of ascites (e.g.,
furosemide) but a great
need remains for improved ascites therapeutics, See Shiozaki, et al. (2006) J.
Physiol. Sci, 56(6):
401-406.
Attention Deficit Hyperactivity Disorder (ADHD)
[0028] ADHD is a problem with inattentiveness, over-activity, impulsivity, or
a combination.
ADHD is the most commonly diagnosed behavioral disorder of childhood. It
affects about 3 - 5% of
school aged children. ADHD is diagnosed much more often in boys than in girls.
ADHD may run in
families, but it is not clear exactly what causes it. Whatever the cause may
be, it seems to be set in
motion early in life as the brain is developing. Depression, lack of sleep,
learning disabilities, tic
disorders, and behavior problems may be confused with, or appear with, ADHD.
Every child
suspected of having ADHD should be carefully examined by a doctor to rule out
possible other
conditions or reasons for the behavior. Most children with ADHD also have at
least one other
developmental or behavioral problem. They may also have a psychiatric problem,
such as depression
or bipolar disorder. ADHD symptoms fall into three groups: lack of attention
(inattentiveness),
hyperactivity, and impulsive behavior (impulsivity). Some children suffer from
primarily
inattentiveness and others have a combination of these symptoms. ADHD is
difficult to diagnosis
but may be identified by a series of developmental, mental, nutritional,
physical, and psychosocial
examination. Attention deficit hyperactivity disorder (ADHD) (2011) PubMed
Health.
[0029] Current treatment of ADHD is a combination of Medications (e.g.,
amphetamine-
dextroamphetamine (ADDERALL)), dexmethylphenidate (FOCALIN), dextraamphetamine
(DEXEDRINE, DEXTROSTAT), lisdexafetamine (Vyvanse), and methylphenidate
(RITALIN) and
behavior therapy. Attention deficit hyperactivity disorder (ADHD) (2011)
PubMed Health. A
recent study suggested that extended-release valproate (EVA), a GABA enhancer,
improved
hyperactivity and impulsivity in an ADHD study. Miyazaki, et al. (2006) Brain
and Development
28(7): 470-472. Also, reduced feedback inhibition by striatal GABA neurons and
intemeurons was
implicated in an animal model of ADHD. Viggiano, et al. (2002) Behavioural
Brain Research
130(1-2): 181-189. However, the current treatment for ADHD leave a great need
for improved
ADHD therapeutics.
Autism Spectrum Disorders (Autism)
[0030] Autism spectrum disorder (ASD) is a range of complex neurodevelopment
disorders,
characterized by social impairments, communication difficulties, and
restricted, repetitive, and
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stereotyped patterns of behavior. Autistic disorder, sometimes called autism
or classical ASD, is the
most severe form of ASD, while other conditions along the spectrum include a
milder form known
as Asperger syndrome, a rare condition called Rett syndrome, and childhood
disintegrative disorder
and pervasive developmental disorder not otherwise specified (usually referred
to as PDD-NOS).
Although ASD varies significantly in character and severity, it occurs in all
ethnic and
socioeconomic groups and affects every age group. Experts estimate that three
to six people out of
every 1,000 may develop ASD. Males are four times more likely to have ASD than
females.
NINDS Autism Fact Sheet (2009).
[0031] Multiple lines of evidence, including genetic and imaging studies,
suggest that the anterior
cingulate cortex and gamma-amino-butyric acid (GABA) system may be affected in
autism.
Compared to controls, the autistic patients show a significant decrease in the
mean density of
GABAA receptors in the supragranular (46.8%) and infragranular (20.2%) layers
of the anterior
cingulate cortex (ACC) and in the density of benzodiazepine binding sites in
the supragranular
(28.9%) and infragranular (16.4%) lamina. In addition, a trend for a decrease
in the density of
benzodiazepine sites was found in the infragranular layers (17.1%) in the
autistic group. These
findings suggest that in the autistic group this down regulation of both
benzodiazepine sites and
GABAA receptors in the ACC may be the result of increased GABA innervation
and/or release
disturbing the delicate excitation/inhibition balance of principal neurons as
well as their output to
key limbic cortical targets. These disturbances may underlie the core
alterations in socio-emotional
behaviors in autism spectrum disorders. Oblak, et al. (August 2009) Autism
Res. 2009
Aug;2(4):205-19. Furthermore, various GABAA subunit types, such as a3 variant
sequences and 04
isoforrns have been linked to autism spectrum disorders. WO 2009/100040. There
is no cure for
ASD, thus there exists a great need for therapeutics to treat autism spectrum
disorders. Therefore,
therapeutics that target the GABAA receptor may be useful in treating autism
spectrum disorders.
Bipolar Disorder
[0032] Bipolar disorder (manic-depressive illness) is a brain disorder that
causes unusual shifts in a
person's mood, energy, and ability to function. They can result in damaged
relationships, poor job
or school performance, and even suicide. About 5.7 million American adults or
about 2.6 percent of
the population age 18 and older have bipolar disorder in any given year.
Bipolar disorder typically
develops in late adolescence or early adulthood. However, some people have
their first symptoms
during childhood, and some develop them late in life. It is often not
recognized as an illness, and
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people may suffer for years before it is properly diagnosed and treated.
National Institute of Mental
Health "Bipolar Disorder" (2008) Complete Publication.
[0033] Bipolar disorder causes dramatic mood swings¨from overly "high" and/or
irritable to sad
and hopeless, and then back again, often with periods of normal mood in
between. Severe changes
in energy and behavior go along with these changes in mood. The periods of
highs and lows are
called episodes of mania and depression. National Institute of Mental Health
"Bipolar Disorder"
(2008) Complete Publication.
[0034] Signs and symptoms of mania (or a manic episode) include: increased
energy, activity, and
restlessness; excessively "high," overly good, euphoric mood; extreme
irritability; racing thoughts
and talking very fast, jumping from one idea to another; distractibility,
difficulty concentrating; little
sleep needed; unrealistic beliefs in one's abilities and powers; poor
judgment; spending sprees; a
lasting period of behavior that is different from usual; increased sexual
drive; drug abuse,
particularly cocaine, alcohol, and sleeping medications; provocative,
intrusive, or aggressive
behavior; and/or denial that anything is wrong. A manic episode is diagnosed
if elevated mood
occurs with three or more of the other symptoms most of the day, nearly every
day, for I week or
longer. National Institute of Mental Health "Bipolar Disorder" (2008) Complete
Publication.
[0035] Signs and symptoms of depression (or a depressive episode) include:
lasting sad, anxious,
or empty mood; feelings of hopelessness or pessimism; feelings of guilt,
worthlessness, or
helplessness; loss of interest or pleasure in activities once enjoyed,
including sex; decreased energy,
a feeling of fatigue or of being "slowed down"; difficulty concentrating,
remembering, making
decisions; restlessness or irritability; sleeping too much, or cannot sleep;
change in appetite and/or
unintended weight loss or gain; chronic pain or other persistent bodily
symptoms that are not caused
by physical illness or injury; and/or thoughts of death or suicide, or suicide
attempts. A depressive
episode is diagnosed if five or more of these symptoms last most of the day,
nearly every day, for a
period of 2 weeks or longer. National Institute of Mental Health "Bipolar
Disorder" (2008)
Complete Publication.
[0036] A mild to moderate level of mania is called hypomania. Hypomania may
feel good to the
person who experiences it and may even be associated with good functioning and
enhanced
productivity. Thus even when family and friends learn to recognize the mood
swings as possible
bipolar disorder, the person may deny that anything is wrong. Without proper
treatment, however,
hypomania can become severe mania in some people or can switch into
depression.
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[0037] In some people, however, symptoms of mania and depression may occur
together in what is
called a mixed bipolar state. Symptoms of a mixed state often include
agitation, trouble sleeping,
significant change in appetite, psychosis, and suicidal thinking. A person may
have a very sad,
hopeless mood while at the same time feeling extremely energized.
[0038] The classic form of the illness, which involves recurrent episodes of
mania and depression,
is called bipolar I disorder. Some people, however, never develop severe mania
but instead
experience milder episodes of hypomania that alternate with depression; this
form of the illness is
called bipolar II disorder. When four or more episodes of illness occur within
a 12-month period, a
person is said to have rapid-cycling bipolar disorder. Some people experience
multiple episodes
within a single week, or even within a single day. Rapid cycling tends to
develop later in the course
of illness and is more common among women than among men.
[0039] Medications known as "mood stabilizers" usually are prescribed to help
control bipolar
disorder (e.g., lithium or valproic acid-DEPAKOTE/VALPROATE). In addition to
medication,
psychosocial treatments¨including certain forms of psychotherapy, are often
used to treat bipolar
disorders. Depending on the medication, side effects include weight gain,
nausea, tremor, reduced
sexual drive or performance, anxiety, hair loss, movement problems, or dry
mouth. Lithium
treatment can cause low thyroid levels, resulting in the need for thyroid
supplementation.
Additionally, Valproate0 may lead to adverse hormone changes in teenage girls
and polycystic
ovary syndrome in women who began taking the medication before age 20.
Further, women
suffering bipolar disorder who wish to conceive, or who become pregnant, face
special challenges
due to the possible harmful effects of existing mood stabilizing medications
on the developing fetus
and the nursing infant. National Institute of Mental Health "Bipolar Disorder"
(2008) Complete
Publication. Improved bipolar disorder therapeutics may be developed that act
to increase GABA
activity.
[0040] Postmortem and genetic studies have linked neuropsychiatric disorders
including
schizophrenia and bipolar disorder with GABAergic neurotransmission and
various specific GABAA
receptor subunits. Further, GABAA receptor-associated proteins involved in
GABAA receptor
trafficking, targeting, clustering, and anchoring that often carry out these
functions in a subtype-
specific manner. Charych, et al. (2009) Neuropharmacology 57(5-6): 481-95.
Therefore, GABAA
receptor specific therapeutics that improve inhibition may be beneficial
because bipolar disease is a
state of alterations of abnormal inhibition/excitation without adequate
inhibition.
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Depression
[0041] Depression is a common but serious illness, the most common are major
depressive
disorder and dysthymic disorder. Major depressive disorder, also called major
depression, is
characterized by a combination of symptoms that interfere with a person's
ability to work, sleep,
study, eat, and enjoy once¨pleasurable activities. Major depression is
disabling and prevents a
person from functioning normally. An episode of major depression may occur
only once in a
person's lifetime, but more often, it recurs throughout a person's life.
National Institute of Mental
Health "Depression" (2008) Complete Publication.
[0042] The forms of depression include:
[0043] Dysthymic disorder, also called dysthymia, is characterized by
long¨term (two years or
longer) but less severe symptoms that may not disable a person but can prevent
one from functioning
normally or feeling well. People with dysthymia may also experience one or
more episodes of major
depression during their lifetimes.
[0044] Psychotic depression, which occurs when a severe depressive illness is
accompanied by
some form of psychosis, such as a break with reality, hallucinations, and
delusions. Postpartum
depression, which is diagnosed if a new mother develops a major depressive
episode within one
month after delivery. It is estimated that 10 to 15 percent of women
experience postpartum
depression after giving birth.
[0045] Seasonal affective disorder (SAD), which is characterized by the onset
of a depressive
illness during the winter months, when there is less natural sunlight. The
depression generally lifts
during spring and summer. SAD may be effectively treated with light therapy,
but nearly half of
those with SAD do not respond to light therapy alone. Antidepressant
medication and
psychotherapy can reduce SAD symptoms, either alone or in combination with
light therapy.
National Institute of Mental Health "Depression" (2008) Complete Publication.
[0046] Depression can be treated with a number of methods. The most common
treatments are
medication and psychotherapy. Antidepressants work to normalize
neurotransmitters, notably
serotonin, norepinephrine, and dopamine. The newest and among the most popular
types of
antidepressant medications are called selective serotonin reuptake inhibitors
(SSRIs). SSRIs include
fluoxetine (Prozac(D), citalopram (Celexa ), sertraline (Zoloft(D), and
several others. Serotonin and
norepinephrine reuptake inhibitors (SNRIs) are similar to SSRIs and include
venlafaxine (Effexor )
and duloxetine (Cymbalta ). SSRIs and SNRIs are more popular than the older
classes of
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antidepressants, such as tricyclics¨named for their chemical structure¨and
monoamine oxidase
inhibitors (MAOIs) because they tend to have fewer side effects. However,
medications affect
everyone differently¨no one¨size¨fits¨all approach to medication exists.
National Institute of
Mental Health "Depression" (2008) Complete Publication.
[0047] For all classes of antidepressants, patients can experience side
effects. Antidepressants may
cause mild and often temporary side effects in some people, but they are
usually not long-term. The
most common side effects associated with SSRIs and SNRIs include: headache,
nausea, insomnia,
nervousness, agitation, and sexual problems. National Institute of Mental
Health "Depression"
(2008) Complete Publication. Tricyclic antidepressants also can cause side
effects including: dry
mouth, constipation, bladder problems, sexual problems, blurred vision, and
daytime drowsiness.
Additionally, patients taking MAOIs must adhere to significant food and
medicinal restrictions to
avoid potentially serious interactions. They must avoid certain foods that
contain high levels of the
chemical tyramine, which is found in many cheeses, wines and pickles, and some
medications
including decongestants. MAOIs interact with tyramine in such a way that may
cause a sharp
increase in blood pressure, which could lead to a stroke. National Institute
of Mental Health
"Depression" (2008) Complete Publication.
[0048] GABA is involved in both clinical depression and in animal models of
depression. Kmm,
et at. (2000) Neuroscience Research 38(2): 193-198. Therefore improved
depression therapeutics
based on the GABAergic system may provide better medication.
Epilepsy
[0049] Epilepsy is characterized by abnormal discharges of cerebral neurons
and is typically
manifested as various types of seizures. Epileptiform activity is identified
with spontaneously
occurring synchronized discharges of neuronal populations that can be measured
using
electrophysiological techniques. Epilepsy is one of the most common
neurological disorders,
affecting about 1% of the population. There are various forms of epilepsy,
including idiopathic,
symptomatic, and cryptogenic. Genetic predisposition is thought to be the
predominant etiologic
factor in idiopathic epilepsy. Symptomatic epilepsy usually develops as a
result of a structural
abnormality in the brain.
[0050] Status epilepticus are a particularly severe form of seizure, which is
manifested as multiple
seizures that persist for a significant length of time, or serial seizures
without any recovery of
consciousness between seizures. The overall mortality rate among adults with
status epilepticus is
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approximately 20 percent. Patients who have a first episode are at substantial
risk for future
episodes and for the development of chronic epilepsy. The frequency of status
epilepticus in the
United States is approximately 150,000 cases per year, with approximately
55,000 deaths being
associated with status epilepticus annually. Sirven and Waterhouse (2003)
American Family
Physician 68: 469-476. Acute processes that are associated with status
epilepticus include
intractable epilepsy, metabolic disturbances (e.g., electrolyte abnormalities,
renal failure, and sepsis),
central nervous system infection (meningitis or encephalitis), stroke,
degenerative diseases, head
trauma, drug toxicity, and hypoxia. The fundamental pathophysiology of status
epilepticus involves
a failure of mechanisms that normally abort an isolated seizure. This failure
can arise from
abnormally persistent, excessive excitation or ineffective recruitment of
inhibition. Studies have
shown that excessive activation of excitatory amino acid receptors can cause
prolonged seizures and
suggest that excitatory amino acids may play a causative role. Status
epilepticus can also be caused
by penicillin and related compounds that antagonize the effects of 7-
aminobutyric acid (GABA).
[0051] Epilepsy is a chronic neurological condition characterized by recurrent
seizures that is
caused by abnormal cerebral nerve cell activity. Epilepsy is classified as
idiopathic or symptomatic.
A nerve cell transmits signals to and from the brain in two ways by (1)
altering the concentrations of
salts (sodium, potassium, calcium) within the cell and (2) releasing chemicals
called
neurotransmitters (e.g. gamma aminobutyric acid, GABA). The change in salt
concentration
conducts the impulse from one end of the nerve cell to the other. At the end,
a neurotransmitter is
released, which carries the impulse to the next nerve cell. Neurotransmitters
either slow down or
stop cell-to-cell communication (called inhibitory neurotransmitters) or
stimulate this process (called
excitatory neurotransmitters). Normally, nerve transmission in the brain
occurs in an orderly way,
allowing a smooth flow of electrical activity. Improper concentration of salts
within the cell and
over activity of either type of neurotransmitter can disrupt orderly nerve
cell transmission and trigger
seizure activity. Certain areas of the brain are more likely than others to be
involved in seizure
activity. The motor cortex, which are responsible for body movement, and the
temporal lobes,
including the hippocampus, which is involved in memory, are particularly
sensitive to biochemical
changes (e.g., decreased oxygen level, metabolic imbalances, infection) that
provoke abnormal brain
cell activity.
[0052] Two molecules regulate cellular chloride levels: KCC2, which transports
chloride out of
cells, and NKCC1, which brings chloride in to the cells. Previous studies in
rats had shown that
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adult nerve cells mostly have KCC2, making their chloride concentrations lower
inside than outside.
Thus, when GABA receptors are activated, chloride tends to come in, with an
inhibitory effect. In
newborn rats, the situation is reversed: their nerve cells mostly have NKCC1,
so chloride is actively
transported inside, making initial chloride concentrations very high. As a
result, GABA activation
causes chloride to exit the cell, with an excitatory effect. See e.g., Cohen
(1981) J. Clin. Pharmacol.
21:537-542; Dzhala, et al. (2005) Nat Med. 11: 1205-1213; Martinez, et al.
(1998) Am. J. Clin.
Nutr. 68:1354S-1357S, the disclosures of each of which is hereby incorporated
by reference in their
entireties. Accordingly, compounds described herein may inhibit seizure
activity in the kainic acid
induced seizure rat model.
[0053] A study demonstrated that NKCC antagonists may help treat seizures in
newborns, which is
difficult to control with existing anticonvulsants. Conventional
anticonvulsants - phenobarbital and
benzodiazepines are ineffective in newborns because their brains are
biochemically different from
adult brains. Conventional anticonvulsants work by mimicking the action of
GABA, a natural
inhibitory chemical in the brain, by activating GABA receptors on the surface
of brain cells. In adult
nerve cells, GABA activation opens up channels that allow chloride to move
into the cell. The cell
thereby acquires a negative charge and becomes less excitable, inhibiting
seizure activity. But in
newborns, chloride is already high, and therefore activating GABA receptors
causes chloride to
move out of nerve cells, creating a paradoxical excitatory reaction that may
actually exacerbate
seizures.
[0054] Traditional anti-epileptic drugs exert their principal effect through
one of three
mechanisms: (a) inhibition of repetitive, high-frequency neuronal firing by
blocking voltage-
dependent sodium channels; (b) potentiation of 7-aminobutyric acid (gamma-
aminobutyric acid,
GABA)-mediated postsynaptic inhibition; and (c) blockade of T-type calcium
channels. Many
current anti-epileptic drug therapies exert their pharmacological effects on
all brain cells, regardless
of their involvement in seizure activity. Common side effects are over-
sedation, dizziness, loss of
memory and liver damage. Furthermore, 20-30% of epilepsy patients are
refractory to current
therapy. Therefore there is a great need for improved epilepsy therapeutics to
reduce both morbidity
and mortality.
Glaucoma
[0055] Glaucoma is a group of diseases that occur when the normal fluid
pressure inside the eyes
slowly rises, damaging the eye's optic nerve and result in vision loss and
blindness. Open-angle
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glaucoma is the most common form and other types include: (1)10w-tension or
normal-tension
glaucoma; (2) angle-closure glaucoma; (3) congenital glaucoma; (4) secondary
glaucomas; and
(5) pigmentary glaucoma including neovascular glaucoma. Glaucoma is usually
detected through a
comprehensive eye exam that includes: (a) visual acuity test; (b) visual field
test; (c) dilated eye
exam; (d) tonometry; and (e) pachymetry. Current glaucoma treatments include
medicines, laser
trabeculoplasty, conventional surgery, or a combination of any of these;
however, there is a great
need for improved glaucoma therapeutics. National Eye Institute Glaucoma Fact
Sheet (2008).
Huntington's Disease
[0056] Huntington's disease (HD) results from neuronal degeneration leading to
uncontrolled
movements, loss of intellectual faculties, and emotional disturbance. HD is an
autosomal dominant
disease caused by a CAG expansion in the Htt gene that leads to a poly-
glutamine expansion in the
disease protein huntingtin. GABAergic interneurons are particularly sensitive
to the accumulation of
mutant huntingtin and die early in the development of HD. Some early symptoms
of HD are mood
swings, depression, irritability or trouble driving, learning new things,
remembering a fact, or
making a decision. As the disease progresses, concentration on intellectual
tasks becomes
increasingly difficult, and the patient may have difficulty feeding himself or
herself and swallowing.
The rate of disease progression and the age of onset vary from person to
person. NINDS Publication
"Huntington's Disease: Hope Through Research" (2009).
[0057] Huntington's disease (HD) is a neurodegenerative disorder that involves
disruptions in
GABA signaling. GABAA is the major inhibitory neurotransmitter in the central
nervous system
(CNS). HD destroys striatal GABAergic neurons. Directing GABAA synthesis,
degradation,
transport, or receptors can control GABA signaling and so drugs that target
these aspects of GABA
metabolism may be used for improved therapeutic treatments for Huntington's
disease. Kleppner
and Tobin (2001) Expert Opin Ther Targets. 5(2):219-39. Physicians prescribe a
number of
medications to help control emotional and movement problems associated with HD
including
tetrabenazine to treat Huntington's chorea (the involuntary writhing
movements). However, the
drugs used to treat the symptoms of HD have side effects such as fatigue,
restlessness, or
hyperexcitability. NINDS Publication "Huntington's Disease: Hope Through
Research" (2009).
Insomnia
[0058] Insomnia is a symptom of sleep disorders, characterized by persistent
difficulty falling
asleep or staying asleep despite the opportunity. NHLBI Diseases and
Conditions Index [Insomnia]
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(2009). Although there are several different degrees of insomnia, three types
of insomnia have been
clearly identified: transient, acute, and chronic. Transient insomnia lasts
from days to weeks. It can
be caused by another disorder, by changes in the sleep environment, by the
timing of sleep, severe
depression, or by stress. Its consequences ¨ sleepiness and impaired
psychomotor performance ¨
are similar to those of sleep deprivation. Acute insomnia is the inability to
consistently sleep well
for a period of between three weeks to six months. Chronic insomnia lasts for
years at a time. It can
be caused by another disorder, or it can be a primary disorder. Its effects
can vary according to its
causes. They might include sleepiness, muscular fatigue, hallucinations,
and/or mental fatigue; but
people with chronic insomnia often show increased alertness. NHLBI Diseases
and Conditions
Index [Insomnia] (2009). Current insomnia drug therapies that target the GABAA
receptor,
hypnotics (e.g., benzodiazepines) may have undesirable side effects, therefore
a great need exists for
improved insomnia therapeutics with reduced side effects.
Ischemia
[0059] Ischemia is a restriction in blood supply, generally due to factors in
the blood vessels, with
resultant damage or dysfunction of tissue due to inadequate oxygenation and
lack of nutrients of the
tissue. Insufficient blood supply causes tissue to become hypoxic, or, if no
oxygen is supplied at all,
anoxic. In contrast with hypoxia, a more general term denoting a shortage of
oxygen (usually a
result of lack of oxygen in the air being breathed), ischemia is an absolute
or relative shortage of the
blood supply to an organ. This can cause necrosis (e.g., cell death). In
aerobic tissues such as heart
and brain, at body temperature necrosis due to ischemia usually takes about 3-
4 hours before
becoming irreversible. Later, more damage occurs due to the accumulation of
metabolic wastes due
to lack of adequate blood supply to the tissue. Complete cessation of
oxygenation of such organs for
more than 20 minutes typically results in irreversible damage.
[0060] Inhibition of NKCC1 activity with bumetanide and furosemide
significantly reduces the
infarct volume and cerebral edema following cerebral focal ischemia suggesting
that NKCC1
antagonists may be useful in treating ischemia. Chen and Sun (2005) Neurol.
Res. 27(3): 280-286.
The typical treatment of ischemia involves "clot-buster" drugs (e.g.,
Alteplase(D) usually given for
stroke and heart attack within this time period. However, restoration of blood
flow after a period of
ischemia can actually be more damaging than the ischemia because
reintroduction of oxygen causes
a greater production of damaging free radicals, resulting in reperfusion
injury, and, eventually,
necrosis. Therefore a great need exists for improved ischemia therapeutics.
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Migraine
[0061] Migraine headaches afflict 10-20% of the U.S. population, with an
estimated loss of 64
million workdays annually. Migraine headache is characterized by pulsating
head pain that is
episodic, unilateral or bilateral, lasting from 4 to 72 hours and often
associated with nausea,
vomiting, and hypersensitivity to light and/or sound. When accompanied by
premonitory symptoms,
such as visual, sensory, speech or motor symptoms, the headache is referred to
as "migraine with
aura," formerly known as classic migraine. When not accompanied by such
symptoms, the
headache is referred to as "migraine without aura," formerly known as common
migraine. Both
types evidence a strong genetic component, and both are three times more
common in women than
men. The precise etiology of migraine has yet to be determined. It has been
theorized that persons
prone to migraine have a reduced threshold for neuronal excitability, possibly
due to reduced activity
of GABA. GABA normally inhibits the activity of the neurotransmitters
serotonin (5-HT) and
glutamate, both of which appear to be involved in migraine attacks. The
excitatory neurotransmitter
glutamate is implicated in an electrical phenomenon called cortical spreading
depression, which can
initiate a migraine attack, while serotonin is implicated in vascular changes
that occur as the
migraine progresses.
[0062] It has been suggested that cortical spreading depression (CSD)
underlies migraines
including migraines with visual aura. It is also believed that CSD underlies
migraine as part of the
trigeminal pain circuit. CSD is characterized by a short burst of intense
depolarization in the
occipital cortex, followed by a wave of neuronal silence and diminished evoked
potentials that
advance anteriorly across the surface of the cerebral cortex. Enhanced
excitability of the occipital-
cortex neurons has been proposed as the basis for CSD. The visual cortex may
have a lower
threshold for excitability and therefore is most prone to CSD. It has been
suggested that
mitochondrial disorders, magnesium deficiency, and abnormality of presynaptic
calcium channels
may be responsible for neuronal hyperexcitability. Welch (1997) Seminars in
Neurobiol. 17: 4.
During a spreading depression event, profound ionic perturbations occur, which
include interstitial
acidification, extracellular potassium accumulation, and redistribution of
sodium and chloride ions to
intracellular compartments. In addition, prolonged glial swelling occurs as a
homeostatic response
to altered ionic extracellular fluid composition, and interstitial
neurotransmitter and fatty acid
accumulation. Studies have shown that furosemide inhibits regenerative
cortical spreading
depression in anaesthetized cats. Read, et at. (1997) Cephalagia 17: 826.
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[0063] Drug therapy is tailored to the severity and frequency of migraine
headaches. For
occasional attacks, acute treatment may be indicated, but for attacks
occurring two or more times per
month, or when attacks greatly impact the patient's daily life, prophylactic
therapy may be indicated.
The side effects of acute and prophylactic treatment agents including
serotonin acting agents, beta-
blockers, tricyclic antidepressants, anticonvulsants, and botulinum toxin type
A injections can limit
their use. GABA modulates nociceptive input to the trigeminocervical complex
mainly through
GABAA receptors. Storer, et al. (2001) Br J Pharmacol. 134(4): 896-904.
Therefore GABAA
receptors may provide a target for the development of new therapeutic agents
for both acute and
prophylactic treatment of headaches including migraines.
Nociceptive pain
[0064] Nociceptive pain occurs in response to the activation of a specific
subset of peripheral
sensory neurons, the nociceptors. Nociceptors are the nerves that sense and
respond to parts of the
body that suffer from damage. They signal tissue irritation, impending injury,
or actual injury. It is
generally acute (with the exception of arthritic pain), self-limiting and
serves a protective biological
function by acting as a warning of on-going tissue damage. When activated,
they transmit pain
signals (via the peripheral nerves as well as the spinal cord) to the brain.
The pain is typically well
localized, constant, and often with an aching or throbbing quality. Examples
include post-operative
pain, sprains, bone fractures, burns, bumps, bruises, inflammation (from an
infection or arthritic
disorder), obstructions, and myofascial pain. Visceral pain is the subtype of
nociceptive pain that
involves the internal organs. It tends to be episodic and poorly localized.
[0065] Nociceptive pain is usually treated with opioids and/or non-steroidal
anti-inflammatory
drugs (NSAIDS) but due to low efficacy, unacceptable, even severe side
effects, and addiction
potential, their use can be limited. GABAA receptors are a target for
therapeutics to treat nociceptive
pain. For example, Hara, et al. (2004) Anesth Analg 98:1380-1384 reports that
the combination of
GABA agonists and L-type calcium channel blockers may be used to reduce
visceral pain.
However, most GABAA agonists are known to have side effects, including
sedation, dizziness,
euphoria, nausea, and blurred vision. Therefore a great need exists for
nociceptive pain therapeutics.
Neuropathic pain
[0066] Neuropathic pain and nociceptive pain differ in their etiology,
pathophysiology, diagnosis,
and treatment. Neuropathic pain is a common type of chronic, non-malignant
pain, which is the
result of an injury or malfunction in the peripheral or central nervous system
and serves no
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protective biological function. It is estimated to affect more than 1.6
million people in the U.S.
population. Neuropathic pain has many different etiologies, and may occur, for
example, due to
trauma, diabetes, infection with herpes zoster (shingles), HIV/AIDS
(peripheral neuropathies), late-
stage cancer, amputation (including mastectomy), carpal tunnel syndrome,
chronic alcohol use,
exposure to radiation, and as an unintended side-effect of neurotoxic
treatment agents, such as
certain anti-HIV and chemotherapeutic drugs.
[0067] In contrast to nociceptive pain, neuropathic pain is frequently
described as "burning,"
"electric," "tingling," or "shooting" in nature. It is often characterized by
chronic allodynia (pain
resulting from a stimulus that does not ordinarily elicit a painful response,
such as light touch) and
hyperalgesia (increased sensitivity to a normally painful stimulus), and may
persist for months or
years beyond the apparent healing of any damaged tissues.
[0068] Neuropathic pain is difficult to treat. Analgesic drugs that are
effective against nociceptive
pain (e.g., opioid narcotics and non-steroidal anti-inflammatory drugs) are
rarely effective against
neuropathic pain. Similarly, drugs that have activity in neuropathic pain are
not usually effective
against nociceptive pain. The standard drugs that have been used to treat
neuropathic pain appear to
often act selectively to relieve certain symptoms but not others in a given
patient (e.g., relief of
allodynia, but not hyperalgesia). Bennett (1998) Hosp. Pract. (Off Ed). 33: 95-
98. Treatment agents
typically employed in the management of neuropathic pain include tricylic
antidepressants (e.g.,
amitriptyline, imipramine, desimipramine, and clomipramine), systemic local
anesthetics, and anti-
epileptic drugs (AED) (e.g., phenytoin, carbamazepine, valproic acid,
clonazepam, gabapentin, and
pregabalin (LYRICAC))). See Lowther (2005) "Pharmacotherapy Update from the
Department of
Pharmacy" Vol. VIII, No. 5. Common side effects include over-sedation,
dizziness, loss of memory
and liver damage. Further, although traditionally not considered useful for
the treatment of
neuropathic pain, recent studies from genetically modified mice indicate that
agents targeting only a
subset of benzodiazepine (GABAA) receptors may provide pronounced
antihyperalgesic activity
against inflammatory and neuropathic pain. Zeilhofer, et al. (2009) J Mol Med
87: 465-469.
Furthermore, in a spinal cord injury (SCI) model of neuropathic pain,
bumetanide, a NKCC1
antagonist, showed an analgesic effect suggesting that normal or elevated
NKCC1 activity plays a
role in the development and maintenance of SCI-induced neuropathic pain.
Cramer, et al. (2008)
Molecular Pain 4: 36. Therefore a great need exists for improved neuropathic
pain therapeutics.
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Postherpetic Neuralgia
[0069] Postherpetic neuralgia is a complication of shingles, a second outbreak
of the varicella-
zoster virus, which initially causes chickenpox. Postherpetic neuralgia
results when nerve fibers are
damaged during an outbreak of shingles. During an initial infection of
chickenpox, some of the
virus remains in the body, lying dormant inside nerve cells. Years later, the
virus may reactivate,
causing shingles. Once reactivated, the virus travels along nerve fibers
causing pain. When the
virus reaches the skin, it produces a rash and blisters. A case of shingles
(herpes zoster) usually
heals within a month. However, these damaged nerves may cause chronic, often
excruciating pain
that may persist for months ¨ or even years ¨ in the area where shingles first
occurred. Some
patients continue to feel pain long after the rash and blisters heal ¨ a type
of pain called
postherpetic neuralgia. A variety of treatments for postherpetic neuralgia
exist, although some do
not experience complete relief from pain.
[0070] This complication of shingles occurs much more frequently in older
adults. About 50
percent of adults older than 60 experience postherpetic neuralgia after
shingles, whereas only 10
percent of all people with shingles do. The symptoms of postherpetic neuralgia
are generally limited
to the area of the skin where the shingles outbreak first occurred including
sharp and jabbing,
burning, or deep and aching pain; extreme sensitivity to touch and temperature
change; itching and
numbness; and headaches. In rare cases, patients might also experience muscle
weakness or
paralysis ¨ if the nerves involved also control muscle movement. A great need
exists for improved
postherpetic neuralgia therapeutics.
Ocular Diseases (e.g., vision disorders, ophthalmic diseases)
[0071] It is estimated that the lifetime costs for all people with vision
impairment who were born in
2000 will total $2.5 billion (2003 dollars). See, generally, Centers for
Disease Control and
Prevention, Economic Costs Associated with Mental Retardation, Cerebral Palsy,
Hearing Loss, &
Vision Impairment, United States, 2003, MMWR (2004) 53: 57-9. These costs
include both direct
and indirect costs. Direct medical costs, such as doctor visits, prescription
drugs, and inpatient
hospital stays, make up 6% of these costs. Direct nonmedical expenses, such as
home modifications
and special education, make up 16% of the costs. Indirect costs, which include
the value of lost
wages when a person dies early, cannot work, or is limited in the amount or
type of work he or she
can do, make up 77% of the costs. These estimates do not include other
expenses, such as hospital
outpatient visits, emergency department visits, and family out-of-pocket
expenses. The actual
22
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
economic costs of vision impairment are, therefore, even higher than what is
generally reported.
U.S. Patent No. 7,251,528.
[0072] Both NKCC and KCC2 are expressed in the outer and inner plexithrm
layers and
colocalized in many putative amacrine cells and in cells of the ganglion cell
layer. However, the
somata of putative horizontal cells displayed only NKCC immunoreactivity and
many bipolar cells
were only immunopositive for KCC2. In the outer retina, application of
bumetanide, a specific
inhibitor of NKCC activity, (1) increased the steady-state extracellular
concentration of K+ ([1(1]0)
and enhanced the light-induced decrease in the [K]0, (2) increased the sPIII
photoreceptor-
dependent component of the ERG, and (3) reduced the extracellular space
volume. In contrast, in
the outer retina, application of furosemide, a specific inhibitor of KCC
activity, decreased sPIII and
the light-induced reduction in [K]0, but had little effect on steady-state
[K]0. In the inner retina,
bumetanide increased the sustained component of the light-induced increase in
[1(]0. These
findings thus indicate that NKCC and KCC2 control the [K]0 and extracellular
space volume in the
retina in addition to regulating GABA- and glycine-mediated synaptic
transmission. In addition, the
anatomical and electrophysiological results together suggest that all of the
major neuronal types in
the retina are influenced by chloride cotransporter activity. Dmitriev, et aL
(2007) Vis Neurosci
24(4): 635-45.
[0073] The bumetanide-sensitive Na+r2C1- cotransporter (NKCC) also clearly
contributes to the
Cr uptake into the pigmented epithelium (PE). This work reinforces the general
consensus that
active secretion of Cr is the major driving force of aqueous humor formation
in mammalian eye and
further substantiates the existence of species differences in the mechanism
that accomplishes
transepithelial Cr transport. Kong, et al. (2006) Invest Ophthalmol Vis Sci.
47(12): 5428-36.
[0074] Additionally, cation-chloride cotransporters are involved in retinal
function by mediating
neural computation in the retina. The directional responses of DS ganglion
cells are mediated in part
by the directional release of gamma-aminobutyric acid from starburst dendrites
and that the
asymmetric distribution of two cotransporters (K Cr cotransporter and Na+K+Cr
cotransporter)
along starburst-cell dendrites mediates direction selectivity. Gavrikov, et
al. (2003) Proc Natl Acad
Sci USA 100(26): 16047-52.
[0075] Further, the function of retina depends on cation chloride transporters
regulating GABA. In
particular, different cation chloride cotransporters in retinal neurons allow
for opposite responses to
GABA. Thus, in the retina, the opposite effects of GABA on different cell
types and on different
23
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
cellular regions are probably primarily determined by the differential
targeting of these two chloride
transporters. See, e.g., Barbour, et al. (May 1991) J Physiol. 436: 169-193;
Keller, et al. (1988)
Pflugers Arch. 411(1): 47-52; and Vardi, et al. (2000) Journal of Neuroscience
20(20): 7657-63.
See, also, Basu, et al. (1998) Invest Ophthalmol Vis Sci. 39(12): 2365-73;
Cia, et al. (2005) J
Neurophysiol. 93(3): 1468-75; Do, et al. (2006) Invest Ophthalmol Vis Sci.
47(6): 2576-82; Hunt,
et al. (2005) Anat Rec A Discov Mol Cell Evol Biol. 287(1): 1051-66; MacLeish
and Nurse (2007)
J Neurophysiol. 98(1): 86-95; Mito, et al. (1993) Am J Physiol. 264(3 Pt 1):
C519-26; Moody
(1984) Annu Rev Neurosci 7: 257-78; Mroz and Lechene (1993) Hear Res. 70(2):
146-50;
Schnetkamp (1980) Biochem Biophys Acta. 598(1): 66-90; and Uhl and Desel
(1989) J Photochem
Photobiol B. 3(4): 549-64.
[0076] Accordingly, a number of vision-threatening disorders of the eye
presently do not have any
effective therapies. One major problem in treatment of such diseases are the
inability to deliver
therapeutic agents into the eye and maintain them there at therapeutically
effective concentrations.
Therefore a great need exists for therapeutics to treat ocular diseases.
Parkinson's Disease
[0077] Parkinson's disease (PD) belongs to a group of conditions called motor
system disorders,
which result from the loss of dopamine-producing brain cells. The four primary
symptoms of PD
are tremor, or trembling in hands, arms, legs, jaw, and face; rigidity, or
stiffness of the limbs and
trunk; bradykinesia, or slowness of movement; and postural instability, or
impaired balance and
coordination. As these symptoms become more pronounced, patients may have
difficulty walking,
talking, or completing other simple tasks. PD usually affects people over the
age of 50. Early
symptoms of PD are subtle and occur gradually. Other symptoms may include
depression and other
emotional changes; difficulty in swallowing, chewing, and speaking; urinary
problems or
constipation; skin problems; and sleep disruptions. NINDS Parkinson's Disease
Information Page
(September 23,2009).
[0078] At present, there is no cure for PD, but a variety of medications
provide dramatic relief
from the symptoms. Usually, patients are given levodopa combined with
carbidopa. Carbidopa
delays the conversion of levodopa into dopamine until it reaches the brain.
Nerve cells can use
levodopa to make dopamine and replenish the brain's dwindling supply. Although
levodopa helps at
least three-quarters of Parkinsonian cases, not all symptoms respond equally
to the drug.
Bradykinesia and rigidity respond best, while tremor may be only marginally
reduced. Problems
24
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
with balance and other symptoms may not be alleviated at all. Anticholinergics
may help control
tremor and rigidity. Other drugs, such as bromocriptine, prannipexole, and
ropinirole, mimic the role
of dopamine in the brain, causing the neurons to react as they would to
dopamine. An antiviral drug,
amantadine, also appears to reduce symptoms. In May 2006, the FDA approved
rasagiline
(AZILECTO) to be used along with levodopa for patients with advanced PD or as
a single-drug
treatment for early PD. NINDS Parkinson's Disease Information Page (2009).
[0079] Parkinson's disease (PD) pathology also disrupts GABA signaling by
destroying the input
from the substantia nigra into striatal GABAergic neurons. Targeting GABA
synthesis, degradation,
transport, or receptors with new therapeutics may control GABA signaling, and
therefore may be
used for improved therapeutic treatments for Parkinson's disease. Kleppner and
Tobin (2001)
Expert Opin. Ther. Targets. 5(2):219-39.
Periodic Leg Movement Disorder
[0080] Periodic limb movement disorder (PLMD) (previously known as nocturnal
myoclonus) is a
sleep disorder where a patient limbs move involuntarily during sleep, and
suffers problems related to
the movement. PLMD differs from restless leg syndrome (RLS) in that RLS occurs
while the
patient is awake as well as when asleep, and when awake, there is a voluntary
response to an
uncomfortable feeling in the legs. In contrast, in PLMD the patient may often
be unaware of the
movements. Cleveland Clinic "Periodic Limb Movement Disorder" (2011).
[0081] Currently, several drugs have been used to treat PLMD including Sinemet
(levodopa),
anticonvulsant medications, benzodiazepines, and narcotics. Although medical
treatment of PLMD
often significantly reduces or eliminates the symptoms of these disorders,
there is no cure for PLMD
and medical treatment must be continued to provide relief. Cleveland Clinic
"Periodic Limb
Movement Disorder" (2011). A recent study found that valproate has a long-term
beneficial effect
on sleep consolidation in patients with PLMD. The principal mechanism of
action of valproate is
believed to be the inhibition of the transamination of GABA (e.g., inhibiting
GABA transaminase,
leads to an increase in GABA). Ehrenberg, et al. (2000) Journal of Clinical
Psychopharmacology
20(5): 574-578.
Restless Leg Syndrome (Restless Leg Disorder)
[0082] Restless leg syndrome (restless leg disorder) is a disorder where
patients suffer from an
urge or need to move the legs to stop unpleasant sensations occurring most
often in middle-aged and
older adults. The cause is not known in most patients but may occur more often
in patients with
25
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
peripheral neuropathy, chronic kidney disease, Parkinson's disease, pregnancy,
iron deficiency, or as
a side-effect of some medications. Restless leg syndrome may result in a
decreased quality of sleep
(insomnia). Many patients may also have rhythmic leg movements during sleep
hours, called
periodic limb movement disorder (PLMD). PubMed Health Website "Restless Leg
Syndrome"
(2011).
[0083] There is no known cure for restless leg syndrome with current treatment
aimed at reducing
stress and muscle relaxation. Pramipexole or ropinirole (), Sinemet, or
tranquilizers (e.g.,
clonazepam) have been used to relieve the symptoms of restless leg syndrome.
PubMed Health
Website "Restless Leg Syndrome" (2011). For example, gabapentin (FANATREX), a
GABA
analogue, has been tested as a treatment for restless leg syndrome with some
promising results.
Imamura & Kushida (2010) Expert Opin Pharmacother. 11(11): 1925-32; See also
Misra, et al.
(2011) Neurology 76(4): 408. Thus, GABA based therapeutics may be useful in
the treatment of
restless leg syndrome (restless leg disorder).
Schizophrenia
[0084] Schizophrenia is a chronic, severe, and disabling brain disorder that
affects about 1.1
percent of the U.S. population age 18 and older in a given year. People with
schizophrenia
sometimes hear voices others do not hear, believe that others are broadcasting
their thoughts to the
world, or become convinced that others are plotting to harm them. These
experiences can make
them fearful and withdrawn and cause difficulties when they try to have
relationships with others.
National Institute of Mental Health "Schizophrenia" website (2008).
[0085] Symptoms usually develop in men in their late teens or early twenties
and women in the
twenties and thirties, but in rare cases, can appear in childhood. They can
include hallucinations,
delusions, disordered thinking, movement disorders, flat affect, social
withdrawal, and cognitive
deficits. No cause of schizophrenia has been determined nor is there any
curative therapy; however,
antipsychotics are used in the treatment of symptoms. National Institute of
Mental Health
"Schizophrenia" website (2008).
[0086] Further, schizophrenia is associated with both decreased numbers and
abnormalities in the
distribution of GABAergic neurons in the cortex, particularly in the cortical
laminae. Kaplan &
Sadock's Comprehensive Textbook of Psychiatry (7th Ed) (2008). In the
postmortem studies of
schizophrenics, antipsychotic naive schizophrenics, and non schizophrenic
controls, show a
significant decrease in the number of GABA containing inter neurons, and a
lessened amount of
26
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
GABA production within these inter neurons in both of the schizophrenic
groups. Nestler (1997)
Nature 385(6617): 578-9. Therefore therapeutic agents that target the GABA
system may be useful
in treating schizophrenia.
Tinnitus
[0087] Tinnitus is the perception of sound within the human ear in the absence
of corresponding
external sound. Tinnitus is not a disease but a symptom resulting from a range
of underlying causes
that can include ear infections, foreign objects or wax in the ear, nose
allergies that prevent (or
induce) fluid drain and cause wax build-up, and injury from loud noises.
Tinnitus can also be caused
by hearing impairment and as a side-effect of some medications. Some cases of
tinnitus are
medically unexplained.
[0088] Tinnitus can be perceived in one or both ears or in the head. It is
usually described as a
ringing noise, but in some patients it takes the form of a high pitched
whining, buzzing, hissing,
screaming, humming, singing or whistling sound, or as ticking, clicking,
roaring, "crickets" or "tree
frogs" or "locusts," tunes, songs, or beeping. It has also been described as a
"whooshing" sound, as
of wind or waves. Tinnitus can be intermittent or it can be continuous in
which case it can be the
cause of great distress. In some individuals, the intensity of tinnitus can be
changed by shoulder,
head, tongue, jaw, or eye movements. To date, no satisfactory therapeutics
exists for tinnitus.
[0089] Partial deafferentation produces a loss of tonic inhibition in the
auditory system that may
lead to inappropriate neuroplastic changes eventually expressed as the
pathophysiology of tinnitus.
The pathological down-regulation of GABA provides a potential mechanism for
this loss of
inhibition. For example, in an animal model of tinnitus, vigabatrin, a GABA
agonist, completely
and reversibly eliminated the psychophysical evidence of tinnitus. Brozoski.
et at. (2007) .1 Assoc
Res Otolaryngol. 8(1): 105-118. Further, the disruption of the N.KCC.1 gene in
mice causes hearing
loss. Kahle, et al. (2004) Proc. Natl. Acad. Sci. USA 102(46): 16783-16788.
Therefore,
therapeutics targeting GABAergic system and/or NKCC1 may be useful in the
treatment of tinnitus.
Withdrawal Syndrome
[0090] Withdrawal syndrome is generally associated with abnormal physical or
psychological
features that follow the abrupt discontinuation of a drug (e.g., medications,
recreational drugs, and/or
alcohol) that has the capability of producing physical dependence. (e.g.,
alcohol withdrawal
syndrome, nicotine withdrawal syndrome, opioid withdrawal syndrome,
benzodiazepine withdrawal
syndrome, methadone withdrawal syndrome, SSR1 discontinuation syndrome,
hydrocodone
27
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
withdrawal syndrome). Common withdrawal symptoms include sweating, tremor,
vomiting,
anxiety, insomnia, and muscle pain. There are different stages of withdrawal.
Generally, a person
will start to feel worse and worse, hit a plateau, and then the symptoms begin
to dissipate. However,
withdrawal from certain drugs (e.g., benzodiazepines, alcohol) can be fatal
and therefore the abrupt
discontinuation of any type of drug is not recommended. Further, many
additions involve
compounds which affect the GABAerigic system (e.g., alcohol and
benzodiazepines.) Therefore,
when a person ceases use of the compound, the GABAergic system is involved in
the symptoms of
withdrawal syndrome. Nutt and Lingford-Hughes (2008) British Journal of
Pharmacology 154(2):
397-405. Therefore agents that act on the GABAergic system may provide
therapeutics to treat
withdrawal syndromes.
[00911 Accordingly, there is a continuing need for compositions and methods
for treatment and/or
prophylaxis of diseases, disorders, and conditions that involve the Na K CF co-
transporters (e.g.,
NKCC1 and NKCC2) including but not limited to addictive disorders, anxiety
disorders, ascites,
attention deficit hyperactivity disorder (ADHD), bipolar disorder, cancer,
endothelial corneal
dystrophy, edema, depression, epilepsy, glaucoma, ischemia, migraine,
neuropathic pain,
nociceptive neuralgia, ocular diseases, pain, postherpetic neuralgia, and
schizophrenia. Additionally,
there is a continuing need for compositions and methods for treatment and/or
prophylaxis of
diseases, disorders, and conditions that involve the GABAA receptors including
but not limited to
Alzheimer's Disease, addictive disorders, anxiety disorders, autism spectrum
disorders (autism),
bipolar disorder, depression, epilepsy, Huntington's Disease, inflammatory
pain, insomnia, migraine,
neuropathic pain, nociceptive pain, pain, Parkinson's disease, periodic limb
movement disorder
(PLMD) (nocturnal myoclonus), personality disorders, psychosis, restless legs
syndrome (RLS),
schizophrenia, seizure disorders, spasticity, tinnitus, and withdrawal
syndromes.
Summary of the Invention
[0092] The present invention provides compounds according to Formulae I, II,
III and IV, which
are arylsulfonamides, including bumetanide derivatives, as provided herein:
[0093] Formula I:
28
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R1
1
0 Z,
R2
R7...6 ... S %.., R4 N
N %).,, I
I µ==== R5 R3
R6
I
or a pharmaceutically acceptable salt thereof,
wherein:
Z is oxygen or nitrogen;
R1 and R2 are each independently hydrogen, alkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl,
heterocycloalkyl, or R1 and R2, together with the atom to which they are
attached, form a 4-7
membered heterocyclic ring that can have one or more additional heteroatoms
and can have one
or more substituents, with the proviso that if Z is oxygen, then R2 is not
present;
R3 and R4 are each independently hydrogen, alkyl, cycloalkyl, cycloalkyl
alkyl, aryl, arylalkyl,
heteroaryl, or heteroarylalky, or R3 and R4, together with the atom to which
they are attached,
form a 4-7 membered heterocyclic ring that can have one or more additional
heteroatoms and
can have one or more substituents;
R5 is alkoxy, halo, aryl, aryloxy, alkaryloxy, arylamino, heteroarylamino,
heterocycloalkyl,
heteroaryl, heteroaryloxy, hetemcycloalkoxy, or alkythio; and
R6 and R7 are each independently hydrogen, acyl, alkyl, cycloalkyl alkyl, aryl
or arylalkyl, or R6
and R7, together with the atom to which they are attached, form a 4-7 membered
heterocyclic
ring that can have one or more additional heteroatoms and can have one or more
substituents.
[09941 Formula II:
Ri
i
....Z 0
R2 R3
1
N,R4
0 140:1
µN
N N=
I 0 R5
R6
II
29
CA 02806664 2013-01-25
WO 2012/018635 PCT/US2011/045390
or a pharmaceutically acceptable salt thereof,
wherein:
Z is oxygen or nitrogen;
R1 and R2 are each independently hydrogen, alkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl,
heterocycloalkyl, or R1 and R2, together with the atom to which they are
attached, form a 4-7
membered heterocyclic ring that can have one or more additional heteroatoms
and can have one
or more substituents, with the proviso that if Z is oxygen, then R2 is not
present;
R3 and R4 are each independently hydrogen, alkyl, cycloalkyl, cycloalkyl
alkyl, aryl, arylalkyl,
heteroaryl, or heteroarylalky, or R3 and R4, together with the atom, to which
they are attached,
form a 4-7 membered heterocyclic ring that can have one or more additional
heteroatoms and
can have one or more substituents;
R5 is halo, aryl, aryloxy, arylamino, heteroarylamino, heterocycloalkyl,
heteroaryl,
heteroaryloxy, heterocycloalkoxy, or alkythio; and
R5 and R7 are each independently hydrogen, acyl, alkyl, cycloalkyl alkyl, aryl
or arylalkyl, or R6
and R7, together with the atom to which they are attached, form a 4-7 membered
heterocyclic
ring that can have one or more additional heteroatoms and can have one or more
substituents.
[0095] Formula III:
Ri
n 9 R8.71
nL.,R2
R6
1111 R4
117 .../S\
db R5 R3
III
or a pharmaceutically acceptable salt thereof,
wherein:
Z is oxygen or nitrogen;
R1 and R2 are each independently hydrogen, alkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl,
heterocycloalkyl, or R1 and R2, together with the atom to which they are
attached, form a 4-7
membered heterocyclic ring that can have one or more additional heteroatoms
and can have one
or more substituents, with the proviso that if Z is oxygen, then R2 is not
present;
30
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WO 2012/018635 PCT/US2011/045390
R3 and R4 are each independently hydrogen, alkyl, cycloalkyl, cycloalkyl
alkyl, aryl, arylalkyl,
heteroaryl, or heteroarylalky, or R3 and R4, together with the atom to which
they are attached,
form a 4-7 membered heterocyclic ring that can have one or more additional
heteroatoms and
can have one or more substituents;
R5 is alkoxy, halo, aryl, aryloxy, alkaryloxy, arylamino, heteroarylamino,
heterocycloalkyl,
heteroaryl, heteroaryloxy, heterocycloalkoxy, or alkythio;
R6 and R7 are each independently hydrogen, acyl, alkyl, cycloalkyl alkyl, aryl
or arylalkyl, or R6
and R7, together with the atom to which they are attached, form a 4-7 membered
heterocyclic
ring that can have one or more additional heteroatoms and can have one or more
substituents;
and
Rg and R9 are each independently hydrogen, alkyl, or Rg and R9 together with
the atom to which
they are attached, form a 3-6 membered substituted or unsubstituted cycloalkyl
or
heterocycloalkyl ring.
[0096] Formula IV:
m R84Ri
H9 .(.._,R2
401
N-R4 R6
R13
N,
R I S0
0 0 195
IV
or a pharmaceutically acceptable salt thereof,
wherein:
Z is oxygen or nitrogen;
R1 and R2 are each independently hydrogen, alkyl, aryl, arylalkyl, heteroaryl,
heteroarylalkyl,
heterocycloalkyl, or R1 and R2, together with the atom to which they are
attached, form a 4-7
membered heterocyclic ring that can have one or more additional heteroatoms
and can have one
or more substituents, with the proviso that if Z is oxygen, then R2 is not
present;
R3 and R4 are each independently hydrogen, alkyl, cycloalkyl, cycloalkyl
alkyl, aryl, arylalkyl,
heteroaryl, or heteroarylalky, or R3 and Rg, together with the atom to which
they are attached,
form a 4-7 membered heterocyclic ring that can have one or more additional
heteroatoms and
can have one or more substituents;
31
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
R5 is alkoxy, halo, aryl, aryloxy, alkaryloxy, arylamino, heteroarylamino,
heterocycloalkyl,
heteroaryl, heteroaryloxy, heterocycloalkoxy, or alkythio;
R6 and R7 are each independently hydrogen, acyl, alkyl, cycloalkyl alkyl, aryl
or arylalkyl, or R6
and R7, together with the atom to which they are attached, form a 4-7 membered
heterocyclic
ring that can have one or more additional heteroatoms and can have one or more
substituents;
and
R8 and R9 are each independently hydrogen, alkyl, or R8 and R9 together with
the atom to which
they are attached, form a 3-6 membered substituted or unsubstituted cycloalkyl
or
heterocycloalkyl ring.
[0097] Embodiments of the present invention provide a pharmaceutical
composition comprising a
compound of Formulae I-IV, a pharmaceutically acceptable salt, solvate,
tautomer, hydrate, or
combination thereof and a pharmaceutically acceptable carrier, excipient, or
diluent. Embodiments
of the present invention provide methods of making the compounds including
compounds described
herein and further provide intermediate compounds formed through the synthetic
methods described
herein to provide the compounds of Formulae I-TV.
[0098] In some embodiments, the arylsulfonamides described herein exclude
furosemide,
bumetanide, and piretanide. In other embodiments, the arylsulfonamides
described herein exclude
one or more compounds disclosed in Examples 1-43 of U.S. Patent Application
Publication No.
2007/0149526. In other embodiments, the arylsulfonamides described herein
exclude one or more
compounds disclosed in Examples 100-136 of WO 2010/085352. In yet other
embodiments, the
arylsulfonamides described herein exclude one or more compounds disclosed in
European Journal of
Medicinal Chemistry (1976) 11(5): 399-406; GB 2207129; Liebigs Annalen der
Chemie (1979) (4):
461-9; American Journal of Physiology (1993) 265(5, Pt. 1): G942¨G954; Journal
of Medicinal
Chemistry (1971) 14(5): 432-9; U.S. Patent No. 4,247,550; U.S. Patent No.
3,985,777; WO
2008/052190; U.S. Patent No. 7,282,519; International Journal of Pharmaceutics
60: 163-169
(1990); U.S. Patent No. 5,073,641; Revista Portugesa de Farmacia 44: 164-169
(1994);
Pharmacology 26: 172-80 (1983); U.S. Patent Application Publication No.
2007/0155729; European
Journal of Pharmacology 344: 269-277 (1998); or JP 49/081334. In still other
embodiments, the
arylsulfonamides described herein exclude one or more compounds of the
formulae:
32
CA 02806664 2013-01-25
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o
o
o
o
0
o
H,N1,..._ /
H2N /
.2N,_ /
-,s
õs
-s
04, 101 OH
0/7' Ill OH
II OH
F
CI
Br
HN.s.,..,..,õ....õ..,....õ,...,õ...õ.õ--
HN.,.,...,..õ....õ......,......,"
H1\1,,,,,...7",,,,...7
0
0
0
H2N.,,,... ,0
0
H2N.,...,, /
0
,s
0// 4111 OH
i/S
OU 0 OH //6
OU a OH
* RN
ONH
0
HN.,,,.........õ....,,,,
HN....,..,....õ..,,.....õ,....
N
H
HN..,,,,_,.......õ,õ...õ,--
0
0
0
0
0
o
,
-s
õN.._ /
H2N /
0ff 10 OH
)
OU 4111 OH
U ; NS
O el OH
OH
Os
HN,,,...õ,,...õ.,---.........õ..,
HN,.......,,,,,.............. o'',.õ..,,..../
HINI,,.......,,,,,...õ/õ..-
0
0
H2N,_ /7
0
0
0
--..'S
H2N,,, i
0
01 is/
N
OH
//OH
H2N,..,.. /7
S
0
/7
OH
0
0 101
rip 10
\ II
HNõ...s.,..,..õ,,,,
HN.......,...õ,,,-..,,,,,,....õ."
H
HN.õ..,....õ,..õ."...õ,-
S".....-
HN.,....õ,,..,,,,,...,...õ
33
CA 02806664 2013-01-25
WO 2012/018635
PCT/US2011/045390
O
o
o
o
H2N,...... /
o
o
H2N, I,/
i
¨0 H2N,...., ,/-"s
iio 40 OH
i
OH 0011 0 OH
=
!ci
o 1110
%lc)N
HN.,..,.....,,,,,,,,,,._,,,,-
H
HN,,,,..õ..õ,,......,,,,,,e
HHN.,.....,õ..õ....õ,,.õ,,....õ.õõ,
0
0
0
0
0
0
H2Nõ.,_ /
?
H2N,,, /
')/S
0// 0 OH I
01/ 0 OH Cl/ 10 OH
0 -0
0
S
I
0
HN,......,.........õ.......õ
0HN.,õ.....õ...,,,..õ.õõ,,.
HNõ.......,_...........õ......,....õ.õ.õ,
0
0
0
0
0
I-12N
0
H2N,.... /
H2N,..._ /
/
//
-.'//S
7
-'S
Olf el OH
OU ell OH
01/ 111 OH
S
S
S
..)
HN..,..õ,.......,,,..õ,..-
.1j
HN,,,,,,..y
HN,õ............õ......-
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1 0 0 0
401 OU 01 OH 0 / 0 S OH
0 0
HN,,, HN
HNõ /0 0 I 0 0
) Nsl
la e 0 OH 1401 1 10 OH
0 0
HNõ,,õ,,, HN-
0 0
a 01 II OH
al 01 I. OH
0
HNõ,,__,,,,..,,,,,,, 0
HN
0 0
0
40 I , 0//) 0 OH
10 OU I. OH
0
HN,,,,,..,., 0
HN
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'SF IP HN / 0 ell 0 H 410 0/7
ell OH 0
',....... //hp ,0
0
0
0
HN....õ,,,...õ,,,,,,,.....õ,õ...,
H
N.,,...,...,,...õ,õ.^..õ....--
H2N
0 ----3 ..-='() HN I
0
OH
0 I. OH 0 H2N s 0 HN,=-
...,.HN .,..,..,.....õ,õ/,.....,õõ,õ.,.,
0
19
0 0,,,,,
HN
OH HN
OH
110 HN.,.........õ,..õ..-...õ.......õ....õ.õ....
10 0H 0 0 HN
õ....,,.,......õ....,...--,,,,.........õ.,..,
Spz--1.,.,...
.\/ HN
0 0
HN
0 OH
0 OH 0
HN,,,..,.......,,,....õ,..õ...=
10 10) 0
HN..õ....,....õ,,,,,,,...-
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N, \
\--1=/ 0 0 0
HN
0 OH IV N 0
OH
0 0
0
Hi\l,,,,,..,.=
P 0
HN
q0
HN
OH
0
1001 4011 OH HN,,,,,,,,,,,,,,,,- li 0 eill
HN
\
( NH
1/ 0
I CNN 0
HN
HN
OH
110 el OH O, = 0H 0
0
0 0
HN 0
HN
OH
0
lel 4111 OH HN li 0 ell
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H
NN
0
0
RN
*
OH
RN
00
=50H
0
HN.,...,,,..,...-..._,,,õ,....
SN
HN,,,....,........
c0
N \
0 HN
0
ell 10 OH ---:M\
RN
1
RN
0
OH
0
110 0
CHt FIN,,,..,..õ.....õ..,"
0
RN
0 OH
HN
OH
1110 0
0 *
0
HN.....,..õõ.
HN.,...,..õ..,,,,....õ/õ.....,.-
H
0.....\\
H
NN,
0
i
FIN
OH
11101 *
HN
1 0 OH
0
111
0
HN.,.....,,,,,...,,...-
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,N
H2N, i ) 0 HN --- % ---
__ / NHN % H2N ,..,., i
0 ,...,,.N N
S ---__ /
N
la OU 01 N
1
0
0 Si 0
HN
H N
0
H2N, i
S
H 2 N , /
H
)
N
. 01 140
0 e
\ /7
0
N--,N
0
HNõ,,,.....,,,,,,,,..,..,,,,,,,
HN.,,,_,,.,....,...õ,õ..,..õ."
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o
,o o
,o
H2N// ,,....
H2N4/
i OH i 0
H2N......
lel 061 . OH . / *
el 0 il OH l
0
0
N.,....õ,....,.....--....õ,,,
0
,0
0
H2N,, //
H2N 9
i
40 04' 10
OF1 0 / OH 0
0
0
NNI
0 0
0 ,0
H2N,.......,/ H2N/
el 0/7 0 OH 40 1 is, OH
0 0
0
0
0
H2N,.,... / 0
,
1
0
S
OH . 0, 0 H2N,.......
OH /I/
I 61
S
III 01 10 OH
0
0
0
...õ....,N,,.....,
HN 0
,
6_,....Ø,
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40I 4101 H2N,õ...../ 0 OH H2N,,,/ 0
H2N if 0
el I a OH op 0/ a
OH
0
yNf 0
0
% i OH
0
H2N-4 0 H2N/ 0
40 1 el OH 10 / 10
OH
0
0 000
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C)
,,
0
H2Nõ.... //
Cl.
0
0
N
2 %.,.., //
0 /
H 0
N .____-- NH
H
if
0110 01 0
N----'
H
0
0
/
.-----
0
H2N,,,/
(----_-1,
0
(.......1
0 0, 0 N
-- --S
H
a 1.1 N&---NH
H
0
0
N
l'i\/-\/
,,0
0
H2N/0
H2N,,, //
(..- .
1
1
N
1
0/1 ill H
01 is N
H
NH
0
0
N
.------.-
0
H2N-40
H2N. //0
110 01 0 H N
1110
1
0 01S 10
__.-----\(-----NFt
0
0
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0 01 I. H2N,47
0 N \.,--/- 0 ou
H2N, /7 iis 0 0
N /
0
0
N
H2N / 0 0
H2N.,
//rA) 0 0
* 0' IS H
a /
H N
0
N"--7 0
...--N--'\,-'.
N
- a H2N, i ) 0
0 H .
H2N, /7 01 * N s 0 0
H
0
0
N,,,,.
N
H2N / 0 0
)0H2N , /
) 0 0
40 1 0 N
H 10 OU *
0
0
"/\/
N
/
/
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H2N, e) 0 0
H2N,,,, i 0 0
. OU 01 HIO 0 / 0 N
Hil)
0
0
140 / 140 H2N,,, e 0 0
,/ a //I H2N,4 0 1. H 0 1 NN 0
I
0
0
N
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0 0 0
N2N , H2N /
la 0's 401 / 1 1-----
\
0 0
N
0 0
H2N/7_,.,,....././ H2N,"
01 1.1 N = 01 le
\
0 0
.N
0
H2N/ iiS H2N j
1 0 N 101 Ofi 0 N
\ \
0 0
0 0 4 0 I,
H2N,"
01 0 1 il N la 0/ 0 N
\ \
0 0
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--- 1 õI H 2N /0 0 (.....-
.7N--"t.--NN H / I
0
N"--t---0
I op 1 lio
1
0
0
N
-----
H2N,,, / 0 0 C:=:-.õ--
.7 H2N,,...... / 0
0
NI,- N__.---N1-1
0 / 0 N\------S
1 101 / 0
1
0
0
,...--N---õ,õõ-------..,,,,,,..---
.------`
,-----
0
0 0/0
i NH
io 01/ ell N i 0
I 1110I Oil el N---(-
1 0
0
'''/\/
N
---"---.
H2Nõ,.... / 0 0
H2N ,......, /
0 0
01 10 N . 101
I
I
;--------i
0
0
N
N
....----"
...-----
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o
0
H2N /0
H2N,..... I
0 0/-'S 0
Nr"---.../c)T--0 0 /IS
e 0 ,..........õ(_s 1
0
0
.....,r4.,.......-..........-
NI/\/
...---""
..-
----
0
H2N ,
0 0
(----)
N".--.--'?'"-3
H2N, /
0 01s I.
1 \N,
101 / 0
Ig =\_----__-N
0
1
0
N
N
-----.---
0 0
0
/S Njil)
H2N,....... / 1
10/ 0 s
.
0 . Nj1:11
1
0
0
,...,,,N,,,,µõ,...õ..-..õ,........õ.õ..,
N
./....?
0 0 JD
112N,,, I
0
/
S
1
el 0 ill
N".----C-)
101 01/ el N
1
0
0
,..",...N.,,.....,",õ...,...-
N
----"--F
.--"--"-'
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H2N.,_ /_,...s0 o
0 0/ s N--.--..--NNO
0 H2N.,_ /
0 0
N -7NI-------0
0
H2N.,_ /-7 0 0
___,.....Nõ......,,...,..,.....õ.õ,õ-
011 10
o
H2N/,....,,, o o
s
N
0 1 1
0 0
N
i
0 off . 1
0
N
.----"---
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0 0
0
H2N /0
H2N.,,../ ,,, H2N/
0
0/S el OH
0/57 010
OH
0/ 1.1 OH
F CI
Br
.....,,,N,,.......,..õ,...,...,,,.,,,"
./..'N'''',../-......./.....
f.N......õõ.........-..õ,....
0
H2N,."
0
,,,.... 0
0
OHH2N/
H2N,...,./
0 0 / 0
OH // 0 OH
fa HN
"...õ,N.,...,,.........,---õ,,,....õ..õ,- OH
e \
N.,,,...,............. \ /
.......õ.N..,..,,,.....õ."..,,,,
0
H2N....."
/2 0 H2N, 1
0
/ $ OH 112N/
0/ 0 OH
0/ 0 OH
0 N ......õõNõ,...,.......õ...õ.........õ," OH 0
0 0 0
82N / 0
H2N./....,.. 0
H2N.. / S
0 / S 0 OH 0
10 OH/ 0
101 OH
(10
N N
õ.....,,N,..,,,õ"...,--......õ,,, NJ
N
ip N HN
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o
o
0
H2N,47
H2N,,,.., /0
H2N...4
0S 41110 / * OH
0/ a OH
/
OH
0
....-'''')
0
S
N
i
0
0
0
H2N,,.....
H2N,4/
H2N.4
/ ell OH
0/ i
0 /
o
OH
0 OH
0-0
0-0
0
N
H
0
H2N,,..... /70
0
0
0
H2N
// ,..,...
H2N.. ,,...s
i
/ 10 OH
6/7 40 OH
0/ 0 OH
S
I
S
S
,,,,N,.........,..,..,..,...õõõ,..-
..)
0
0
H2N,,,17
H2N,...."
0,7 0 OH
0/ 0 OH
S
S
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a ou 0 HN I ,// 1 ,9
0 Hla/0 HN
,,...... / 0 0 OH
0 N
0 N
-------'
HN / ,,..._ --S ,0
OH III 01 el 0 N / s 0
0 0 OH 01 1111
0
0
1\1
N
-------'
0 01 ell OH /0
0
N s// ,0 0
OH
0 1\1
0
N
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0
0
--s
,
0
,
0 OU lel OH
1
of/ 0 OH
0
N
/
/
0
0
,,.,, I
s
0
0
110
HNe
1 0 OH
i
00, i 0 OH
0
0
N
.---".-N''',-----.-
1
H2N
0
I
HN
0
HNe
0
0 0 OH
I
0 0,/ 0 OH
0
0
N
..----'
/
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0
H2N OH e 0
HN
0 411 0
OH
0 ell
, N
1------'
./..."'
H isr-----...3.
\ NIV 0 07-:-4..---._1____
HN 0 H .\ .* HN
0
OH
la el 0
N 10 el 0
'''/\/
....-----
/
sn.
HN 1111
0
HN
0 0 OH 0
OH
.."", N ..,..õ,.....,..õ,õ....õ,õ,.--II 0 4111
N
.-"."---j
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N/) 0 0
0
\-t N
HN OH
la 0 0 OH 0 1. 0
N
/ /
2 0 0
HN q HN
0 0
0 IS OR 10 IS OH
N.,,,,.,,,,,-,,,,7 e,,,,N,,,,,,,,,=s,,,,,,,,
( \NH
I / 0 CNHIJ 0
RN
HNI0
OH
0 Si OH 0 10 0
N/\/
/
/
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0 HN 0
OH 0 0 HN
0 0 1401
101 IS OH 0
"/\/
H
ON
cNN 1 -7T---\ HN 0
0 HN
0 10 OH 0 N
10 Si OH 0
N
0 SN
\ -7----'\ HN 0
0----\
I HN 0
I. 0 OH 0
40 0
OH .1
N
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=N C)----A HN
0
1101 11011 OH 0 N 1110 el OH 0
N
r \\IN H czN H
HN \ -7----\ HN
0
0 0 OH 0 Nõ....---"N Ill IS OH 0
-,"--.-
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_,...N
,0 FIN"- % ,0
1 -----.. /7 1
el 0// 14111
Cci 0 N
0 0
_...,,,N...,........,...õ.õ,,õ,..õ....,
N
,0 0
H2N,......., H2N,......, ,
H
N \
0 oe . 0 0,/i 0
\ /IN
N----ff
0 0
l'q i'l
,0 0 õ0 0
H2N,....... // H2N,...., 7/
0"......"-
C(i/ 101
0 100 0
10 ...,,...,,,,.......N.,,,..._õ.õ/
0 0
N '''
.----- /
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0
0
H2N , ,-oo H2N, ,0
,
o
, iso0 0 /
oso0.
o
,,,,õN.,,,,,....,...--,.....=
/N
7 7
0 0
,0 0 0
H2N i
142N, //
-''S
411 * oi * o 0
01 lo o
I
0
o
N,,..,õõ......"..,,,-
.====N-.'",,/
0 0
0
H2N, g
,0 0
)
I-12N -,..,.., g
0/1 10 (D 0
l / II
L, 0
I
1
o
õ,....õNõ..,..,,,,...--,,,...".õ,...-
...õ,...õN.,..,..,õ,...õ,-.....,
7
7
[00991 In still other embodiments, In still other embodiments, the
sulfonamides of the Formulae I
and II exclude one or more compounds of the formulae:
58
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--..õ.
......õ
4111 0
0 10110
FIN 0
FIN 10
HN
01 ,...-0
?--N-CH3 ? zIN -CH 3 0
H3C-O 6 H H3C-0
6 6H3 H-0 6 H
NH ./N....,"=.NH
0 =
HN 0 IIP
0/%1
Lõ,, N 0 ,...,=õ0 uir ce, 0
L.,
r I NH N H2
0 0 9 0 2
LLNH4
"e.."*.%""****-NNH
0
0
H lit
r---N---..-N 40 40
%-,0 11110 0 ._
..'N H
....N.%) 0 0 2 11-N H2
0 0
H
H 0 011' H # 0 li
i0
, g-NH2
04,i_ j =
C.) = gNH3
0
0
Lis NH
.ee 1 H 110)
'-µ..
8 NH2 la-- 0 0
i
,
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LI NHr, 0
LI
,,,,
NH N.
(Xs., 0
1.11 Q, 0 I H
It NH2 .."=Ni N 0 ....0
10:: it
NH2
0 0
LINN
IA.
H
04111
4111)
I H 0H ilo
I NH2
H2N --"N===.,--N''...) =
0 0rNt-I2
L1 --..NH 01
\----\-----.NH11111
H I..0N
0
IN...,,..N ...:
If NH2 40 F4 40
eo
0 0
f/ NH
0 0 2 ,
\-----\--..NH\----\---NH
\----\-----
NH
41 0 0
H S0 0
01 1/0 H a 0
=-=,....,õ,,N 8
-
...õ..4_,...N 8
,p., ,
0 0 P-'4NH 2
S 0 // NI-12 S
0 u ''NH 2
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ri___I
0 HN 0 CI HHN 0 CI
H N
N
WNH02
--- n 0 6 NH2 ) 0
*----. .N 113C,
(0 1 \
=
a 0 NH
=) CICH3 O., = 1 0,
Fr g i H3o...1\r g ,
H3 H3 d
ri----$ ro
0 0
a 0 NH 1 CI 0 NH
I I
i 0 0
0
H3C l
p
p 0
0
c 0 NH
CI 0 NH
0,,, I OH
I13,;. 0`CH3 N" I
II HO 0
H 0 0
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0 0
CI 0 NH
HN 40 NH
0.z., I OH
H 0 ,,.0
........,,,....7--.N R 0
0
8 NH2
H 0
,
0 0,g
H2N
li 00 OCH3
0 1
ro
H CI
CI 0 NH 0
0,7. I
b
1-IN--R 0 0 0"...õ..,.../.\ o
0
C! NH
a 401 NH
0,..7. I
0......õ.......õ,"+õ............,.."
H2N1
o.7., H2NI
0 0
0
0
SR_
NH
NH
NH
o 141111
o 141111
0 =
H 0 0 40 11 40 iio //
=-=.,..õ...õ,N H 0 0 //
S.,,,.
S
0 0 1/ NH2
0 0 2
S 0 NH2
NH
0 01111
le fl io i
S 0 2
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[0100] The compounds of the present invention antagonize NKCC1 and/or GABAA
receptors. The
compounds of the present invention are useful in the treatment of conditions
that involve NKCC1
and/or GABAA receptors. In a preferred embodiment, these compounds are
selective antagonists of
NKCC1 and/or GABAA receptors. In a preferred embodiment, these compounds are
selective
antagonists of GABAA receptors. In a preferred embodiment, these compounds are
selective
antagonists of GABAA receptors comprising an o'4, a5, or a6 subunit.
Methods of Use
[0101] In another embodiment, the invention relates to a method for treating
addictive disorders,
Alzheimer's Disease, anxiety disorders, ascites, attention deficit
hyperactivity disorder (ADHD),
autism spectrum disorders (autism), bipolar disorder, cancer, cognitive
function (e.g., cognitive
impairment, cognitive dysfunction), depression, edema, endothelial corneal
dystrophy, epilepsy,
glaucoma, Huntington's Disease, inflammatory pain, insomnia, ischemia,
migraine with aura,
migraine, migraine without aura, neuropathic pain, nociceptive neuralgia,
nociceptive pain, ocular
diseases, pain, Parkinson's disease, periodic limb movement disorder (PLMD)
(nocturnal
myoclonus), personality disorders, postherpetic neuralgia, psychosis, restless
legs syndrome (RLS),
schizophrenia, seizure disorders, spasticity, tinnitus, or withdrawal
syndromes comprising
administering an effective amount of a compound of Formulae I, II, III or IV:
[0102] In another aspect, the invention relates to a method of inhibiting the
Na K CF cotransporters
comprising administering an effective amount of a compound of Formulae I, II,
III or IV.
[0103] In yet another aspect, the invention relates to a method of inhibiting
the NKCC1 (CCC I,
BSC2) isoform of the Na+K+Cl- cotransporters comprising administering an
effective amount of a
compound of the formula I, II, III or IV. In still another aspect, the
invention relates to a method of
inhibiting the NKCC2 (CCC2, BSC1) isoform of the NerCI- cotransporters
comprising
administering an effective amount of a compound of the formula f, H, III or
IV. In another aspect,
the invention relates to a method of inhibiting both the NKCC1 (CCC1, BSC2)
isoform and the
NKCC2 (CCC2, BSC I) isoform of the NeK+Cl- cotransporters comprising
administering an
effective amount of a compound of the formula I, II, III or IV.
[0104] The present invention also provides methods of using the compounds of
Formulae I-TV for
treating disorders involving the NerCI- co-transporters including but not
limited to addictive
disorders (e.g., compulsive disorders, eating disorders (e.g., obesity),
addiction to narcotics/physical
dependence, alcohol addiction, narcotic addiction, cocaine addiction, heroin
addiction, opiate
63
WO 2012/018635 CA 02806664 2013-01-25PCT/US2011/045390
addiction, alcoholism, and smoking); anxiety disorders (e.g., anxiety, acute
anxiety, panic disorder,
social anxiety disorder, obsessive compulsive disorder (OCD), panic disorder,
panic symptoms,
post-traumatic stress disorder (PTSD), generalized anxiety disorder, and
specific phobia); ascites
(e.g., peritoneal cavity fluid, peritoneal fluid excess, hydroperitoneum,
abdominal dropsy, cancer
related to ascites, tumors related to ascites); attention deficit
hyperactivity disorder (ADHD); bipolar
disorder (e.g., manic-depressive illness, manic phase, depressive phase, mixed
bipolar state, bipolar I
disorder, bipolar II disorder, rapid-cycling bipolar disorder); cancer (e.g.,
tumors, cancer related to
ascites, tumors related to ascites); depression (e.g., psychotic depression,
postpartum depression,
seasonal affective disorder (SAD), cortical spreading depression, dysthymia
(mild depression));
edema (e.g., central nervous system edema); endothelial corneal dystrophy
(e.g., post-chamber
ocular diseases); epilepsy (e.g., seizures, epileptic seizures, a seizure
cluster, an acute seizure (e.g.,
status epilepticus), seizure disorder, and other neurological disorders
involving seizures (e.g.,
cerebral palsy, Ohtahara Syndrome)); glaucoma (e.g., increased intraocular
pressure, angle-closure
glaucoma, neovascular glaucoma, open-angle glaucoma); ischemia (e.g., cardiac
ischemia
(myocardial ischemia), intestinal ischemia, mesenteric artery ischemia (acute
mesenteric ischemia),
hepatic ischemia, and cerebral ischemia (brain ischemia)); migraine (e.g.,
migraine including
headache, migraine variant, migraine headache, cervical migraine syndrome,
acute confusional
migraine, migraine with aura, migraine without aura); neuropathic pain (e.g.,
diabetic neuropathy,
nerve injury, nerve tract injury, neuropathic pain associated with visceral
and/or somatic pain,
peripheral neuropathy, chemotherapy-induced neuropathy, chemotherapy-induced
peripheral
neuropathy, neuralgia, polyneuropathy, mononeuropathy, mononeuritis multiplex,
autonomic
neuropathy, symmetrical peripheral neuropathy, radiculopathy, large fiber
peripheral neuropathy,
small fiber peripheral neuropathy, idiopathic neuropathic pain); nociceptive
neuralgia; ocular
diseases (e.g., diseases of retina-retinal detachment and injury response;
diseases of electrical
transmission between various retinal elements such as rods, cones, amacrine
and horizontal cells,
activity of retinal ganglion cells, dysfunction of MilIler (glial) cells,
abnormal function of the retinal
pigment epithelium; dysfunction of formation of the retina in development and
the appropriate
maintenance of neural connections following maturation and development;
regulation of normal
electrolyte homeostasis in various chorioretinal and vitreoretinal diseases;
abnormal function of
Muller cells in diabetic retinopathy; loss of normal electrical activity in
degenerative diseases of
retina, inherited and those of unknown etiology; inflammatory diseases and
conditions of the eye
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such as chorioretinitis, multiple sclerosis; infectious processes in the eye
with abnormal
inflammatory and injury responses; uveitis; abnormal function of Miller cells
of retina and disease
thereof; dysfunction of RPE-retinal pigment epithelium (e.g., diseases of
RPE); endothelial
(posterior) corneal dystrophies, which result from primary endothelial
dysfunction, (e.g., Fuchs
endothelial corneal dystrophy (FECD), posterior polymorphous corneal dystrophy
(PPCD) and
congenital hereditary endothelial dystrophy (CHED)); retinitis pigmentosa; age-
related macular
degeneration (e.g., dry age-related macular degeneration, exudative age-
related macular
degeneration, and myopic degeneration); retinopathy (e.g., diabetic
retinopathy, proliferative
vitreoretinopathy, and toxic retinopathy) and diseases of aqueous humor
formation (e.g., glaucoma));
pain (e.g., chronic inflammatory pain, chronic musculoskeletal pain, pain
associated with arthritis,
pain associated to osteoarthritis, fibromyalgia, back pain, bone pain
associated with cancer, cancer-
associated pain, chemotherapy-induced neuropathy, chemotherapy-induced
peripheral neuropathy,
H1V-treatment induced neuropathy, H1V-treatment induced neuralgia, pain
associated with digestive
disease, pain associated with Crohn's disease, pain associated with autoimmune
disease, pain
associated with endocrine disease, pain associated with diabetic neuropathy,
pain associated with
shingles or herpes zoster, phantom limb pain, spontaneous pain, chronic post-
surgical pain, chronic
temporomandibular pain, causalgia, postherpetic neuralgia, AIDS-related pain,
complex regional
pain syndromes type I and II, trigeminal neuralgia, chronic back pain, pain
associated with spinal
cord injury and/or recurrent acute pain); postherpetic neuralgia (e.g.,
shingles, herpes zoster); and
schizophrenia. In a preferred embodiment, these compounds are selective
antagonists of NKCC1.
[0105] The present invention also provides methods of using the compounds of
Formulae I-IV for
treating disorders involving a GABAA receptor including but not limited to
Alzheimer's Disease
(AD), addictive disorders (e.g., compulsive disorders, eating disorders (e.g.,
obesity, anorexia
nervosa, bulimia), addiction to narcotics/physical dependence, alcohol
addiction, narcotic addiction,
cocaine addiction, heroin addiction, opiate addiction, alcoholism, and
smoking); anxiety disorders
(e.g., anxiety, acute anxiety, panic disorder, social anxiety disorder,
obsessive compulsive disorder
(OCD), panic disorder, panic symptoms, post-traumatic stress disorder (PTSD),
generalized anxiety
disorder, and specific phobia); autism spectrum disorders (autism); bipolar
disorder (e.g., manic-
depressive illness, manic phase, depressive phase, mixed bipolar state,
bipolar I disorder, bipolar II
disorder, rapid-cycling bipolar disorder, bipolar I disorder, bipolar II
disorder); depression (e.g.,
psychotic depression, postpartum depression, seasonal affective disorder
(SAD), cortical spreading
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depression, dysthyrnia (mild depression)); epilepsy (e.g., seizures, epileptic
seizures, a seizure
cluster, an acute seizure (e.g., status epilepticus), seizure disorder, and
other neurological disorders
involving seizures (e.g., cerebral palsy, Ohtahara Syndrome)); Huntington's
Disease (HD) (e.g.,
Huntington's chorea); insomnia, migraine (e.g., migraine including headache,
migraine variant,
migraine headache, cervical migraine syndrome, acute confusional migraine,
migraine with aura,
migraine without aura, chronic migraine, transformed migraine); neuropathic
pain (e.g., diabetic
neuropathy, cluster headache, nerve injury, nerve tract injury, neuropathic
pain associated with
visceral and/or somatic pain, peripheral neuropathy, chemotherapy-induced
neuropathy,
chemotherapy-induced peripheral neuropathy, HIV-treatment induced neuropathy,
HIV-treatment
induced neuralgia, neuralgia, polyneuropathy, mononeuropathy, mononeuritis
multiplex, autonomic
neuropathy, symmetrical peripheral neuropathy, radiculopathy, large fiber
peripheral neuropathy,
small fiber peripheral neuropathy, idiopathic neuropathic pain); nociceptive
pain; pain (e.g., acute
pain, acute inflammatory pain, chronic inflammatory pain, chronic
musculoskeletal pain, pain
associated with arthritis, pain associated to osteoarthritis, fibromyalgia,
back pain, bone pain
associated with cancer, cancer-associated pain, chemotherapy-induced
neuropathy, chemotherapy-
induced peripheral neuropathy, pain associated with digestive disease, pain
associated with Crohn's
disease, pain associated with autoimmune disease, pain associated with
endocrine disease, pain
associated with diabetic neuropathy, pain associated with shingles or herpes
zoster, phantom limb
pain, spontaneous pain, chronic post-surgical pain, chronic temporomandibular
pain, causalgia,
postherpetic neuralgia, AIDS-related pain, complex regional pain syndromes
type I and II,
trigeminal neuralgia, chronic back pain, pain associated with spinal cord
injury, incisional post
operative, trauma associated, burns, recurrent acute pain, head pain,
headache, nonmigrainous,
specific non-migraine head pains, tic dolureaux, postherpetic neuralgia, ice
pick headache);
Parkinson's disease, periodic limb movement disorder (PLMD) (nocturnal
myoclonus), personality
disorders, psychosis, restless legs syndrome (RLS), seizure disorders,
personality disorders,
schizophrenia, tinnitus, and withdrawal syndromes (e.g., alcohol withdrawal
syndrome, nicotine
withdrawal syndrome, opioid withdrawal syndrome, benzodiazepine withdrawal
syndrome,
methadone withdrawal syndrome, SSRI discontinuation syndrome, hydrocodone
withdrawal
syndrome, cocaine withdrawal syndrome, heroin withdrawal syndrome).
[0106] The present invention further provides methods for treating a patient
diagnosed with risk
factors for a condition selected from the group consisting of addictive
disorders, Alzheimer's
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Disease, anxiety disorders, ascites, attention deficit hyperactivity disorder
(ADHD), autism spectrum
disorders (autism), bipolar disorder, cancer, cognitive function (e.g.,
cognitive impairment, cognitive
dysfunction), depression, endothelial corneal dystrophy, edema, epilepsy,
glaucoma, Huntington's
Disease, inflammatory pain, insomnia, ischemia, migraine, migraine with aura,
migraine without
aura, neuropathic pain, nociceptive neuralgia, nociceptive pain, ocular
diseases, pain, Parkinson's
disease, periodic limb movement disorder (PLMD) (nocturnal myoclonus),
personality disorders,
postherpetic neuralgia, psychosis, restless legs syndrome (RLS),
schizophrenia, seizure disorders,
spasticity, tinnitus, and withdrawal syndromes comprising administering an
effective amount of a
compound of Formulae I, II, III, or IV.
[0107] Embodiments of the present invention provide kits including the
compounds including
compounds described herein. These kits may be used in the treatment methods
disclosed herein. In
another embodiment, the kits may include instructions, directions, labels,
warnings, or information
pamphlets.
[0108] Embodiments of the present invention provide uses of the compounds
described herein for
the preparation of a medicament for carrying out the aforementioned utilities.
[0109] In a preferred embodiment, compounds described herein show differential
activity with
stronger effect on the central nervous system and less diuretic effects. For
example, compounds
described herein may be used in long-term (maintenance) therapy without
significant diuretic effect.
Also, the arylsulfonamides described herein may be used in combination therapy
with diuretics
because of their lack of diuretic effect. Additionally, compounds described
herein do not interfere
with diuretics or cause severe side effects when administered in conjunction
with or concurrently
with a diuretic.
[0110] In another embodiment, the compounds described herein may be
administered in
combination with a second agent.
Enhanced Permeability
[0111] Embodiments of the present invention provide compounds capable of
passage across the
blood-brain barrier comprising a compound of Formulae I-TV, or a
pharmaceutically acceptable salt,
solvate, tautomer or hydrate thereof. In some embodiments, compounds of the
present invention
may have increased lipophilicity and/or reduced diuretic effects compared to
the diuretic or diuretic-
like compounds. The lipiphilicity can be measured by determining the
hydrophile-lipophile balance
(HLB) or the partition coefficient (e.g., the distribution of a compound
between water and octanol).
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In further embodiments, compounds of the present invention may result in fewer
undesirable side
effects when employed in the regulatory, (i.e., preventive, management),
and/or treatment, methods
described herein. In a preferred embodiment, compounds described herein show
improved CNS
pharmacologic properties and increased transit across the blood-brain barrier
(BBB).
Weak Diuretic Effect
[0112] In some embodiments, the level of diuresis that occurs following
administration of an
effective amount of a compound provided herein as Formulae I-IV, is less than
about 99%, 90%,
80%, 70%, 60%, 50%, 40%, 30%, 20% or 10% of that which occurs following
administration of an
comparable amount of a diuretic compound (e.g., bumetanide, furosemide,
piratanide, torsemide,
azosemide). For example, the compound may have less of a diuretic effect than
a diuretic compound
(e.g., bumetanide, furosemide, piratanide, torsemide, azosemide) when
administered at the same
mg/kg dose.
Selective Activity
Na+ Cr co-transporters
[0113] The compounds of the present invention of Formulae 1-TV described
herein may be used for
the regulation, including prevention, management and treatment, of a range of
conditions including,
but not limited to disorders that involve at least one of the Na+Kitl- co-
transporters (e.g., NKCC1,
KNCC2) or K*C1- co-transporters (e.g., KCC1, KCC2, KCC3, KCC4).
[0114] In a preferred embodiment, the invention comprises a method of
inhibiting basolateral
bumetanide-sensitive Na+K C1- cotransporters (e.g., NKCC1) comprising
administering a
composition comprising a compound described herein, wherein the inhibition of
apical bumetanide-
sensitive Na+K+Cl- cotransporters (e.g., NKCC2) is no more than 10%, 15%, 25%,
or 50% of the
effect on basolateral bumetanide-sensitive Na+K+Cr cotransporters (e.g.,
NKCC1). In yet another
embodiment, the invention comprises a method of inhibiting apical bumetanide-
sensitive Na+IMI-
cotransporters (e.g., NKCC2) comprising administering a composition comprising
a compound
described herein, wherein the inhibition of basolateral bumetanide-sensitive
Na+K+Cl- cotransporters
(e.g., NKCC1) is no more than 10%, 15%, 25%, or 50% of the effect on apical
bumetanide-sensitive
Na+KFC1- cotransporters (e.g., NKCC2). Some preferred compounds described
herein may not act
on the GABAA receptor or show only minimal activity on GABAA receptors.
GABAA receptors
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[0115] Compounds of the present invention of Formulae I-TV described herein
may be used for the
regulation, including prevention, management and treatment, of a range of
conditions including, but
not limited to disorders that involve at least one of the GABAA receptor.
[0116] In another embodiment, compounds described herein may show selective
effect on a subset
of GABAA receptors in the CNS and less of the side-effects usually associated
with agents that act
on GABAA receptors. For example, compounds described herein exhibit less
sedation and less
suppression of respiration, cognition, or motor function. In another
embodiment, compounds
described herein may show a selective effect on GABAA receptors comprising an
a5 subunit or an a6
subunit. In another embodiment, compounds described herein show a selective
effect on GABAA
receptors comprising an a4 subunit.
[0117] In one embodiment, the invention comprises a method for antagonizing
parasynaptic (herein
defined as pre- or extra-synaptic) GABAA receptors comprising administering a
composition
comprising an effective amount of a compound of the Formulae I-TV or a
pharmaceutically
acceptable salt thereof. In another embodiment, the invention comprises a
method for antagonizing
parasynaptic GABAA receptors comprising an a4, as, or a6 subunit comprising
administering a
composition comprising an effective amount of a compound of the Formulae I-TV
or a
pharmaceutically acceptable salt thereof.
[0118] Compounds described herein may have antagonistic effects on GABAA
receptors located
parasynaptically. In one embodiment, compounds described herein may have
antagonistic effects on
GABAA receptors comprising an a4, as, or 0(.6 subunit located
parasynaptically. In another
embodiment, the invention comprises a method for antagonizing parasynaptic
GABAA receptors
comprising an a4, a5, or a6 subunit comprising administering a composition
comprising a compound
described herein, wherein the antagonism of GABAA receptors with an al, a2, or
a3 subunit is no
more than 10%, 15%, 25%, or 50% of the effect on a GABAA receptor with an a4,
as, or a6 subunit.
[0119] Compounds described herein may preferentially bind GABAA receptors. In
one
embodiment, compounds described herein may preferentially bind GABAA receptors
comprising an
al, az, a3, a4, as, or a6 subunit. Preferential binding of the compounds of
this invention may be
reflected in the effective concentration (ECso), e., the concentration of the
compound in vitro at
which the antagonist effect is half the maximal antagonism demonstrated by the
respective
compound on the particular receptor. In particular, more preferred compounds
of this invention will
be those whose EC50 for GABAA receptors with an a4, a5, or a6 subunit are no
more than 10%,
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15%, 25%, or 50% of the EC50 of the same compound for GABAA receptors having
an al, a2, or a3
subunit.
[0120] Compounds described herein are effective in humans and animals to
decrease seizures,
decrease pain responses, and decrease migraine in humans and animal models.
For example,
compounds described herein may preferentially bind to GABAA receptor subtypes
and have an
antagonistic effect on GABAA receptors that is different from classic
benzodiazepine and barbiturate
mechanisms. Compounds described herein may not act on the Ner2C1-
cotransporter (NKCC1 or
NKCC2). Unlike a diuretic compound (e.g., bumetanide, furosemide, piretanide,
azosemide, and
torsemide), compounds described herein may not elicit diuresis. For example,
compounds described
herein may not increase urine output, sodium excretion, or potassium
excretion.
[0121] The foregoing and other objects and aspects of the present invention
are explained in greater
detail in reference to the drawing and description set forth herein.
Brief Description of the Drawings
[0122] FIGURE 1 is a schematic illustration of a possible mechanism for the
action of compounds
described herein that selectively antagonize parasynaptic GABAA receptor
isoforms in GABAergic
interneurons. In this suggested mechanism, (1) GABA is released from the pre-
synaptic terminal by
activated inhibitory neurons, (2) GABA binds to post-synaptic GABAA receptors
that activates them
thereby increasing inhibition (e.g., hyperpolarization of the post-synaptic
neuron), (3) GABA also
binds to parasynaptic (e.g., presynaptic and extrasynaptic) GABAA receptors,
(4) in one possible
mechanism of action, compounds described herein selectively binds to
parasynaptic at variant
GABAA receptors (inhibiting the negative feedback loop), thus increasing GABA
release. This leads
to the restoration of the balance of excitation and inhibition by increasing
the inhibitory stimulus
applied to post-synaptic neurons.
[0123] FIGURE 2 illustrates the activating effect of I M clobazam, 0.1 M
zolpidern, and 1 M
diazepam on the current in al containing GABAA receptor isoforms at 10 p.M
GABA.
[0124] FIGURE 3 illustrates the inhibiting effect of 10 M select compounds on
the current in a1,
a4, las, and a6 containing GABAA receptor isoforms at 10 M GABA.
[0125] FIGURE 4 illustrates the inhibitory effect of select compounds at 10 M
concentration on
GABAA receptor activity in the presence of 10 M GABA. 100% represents full
activation of a
GABAA receptor. 50% to 20% value represents strong inhibition of GABAA
receptor activity.
Bumetanide (BTX) was used as a negative control.
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[0126] FIGURE 5 illustrate the inhibitory effect of 10 pt.N4 of select
compounds on a4133721, GABAA
receptor isoform activity in the presence of 10 tM GABA. 100% represents full
activation of the
GABAA receptor. 50% to 20% value represents strong inhibition of GABAA
receptor activity.
Bumetanide (BTN) was used as a negative control.
[0127] FIGURE 6A-D illustrate the inhibitory effect of 10 [tM of select
compounds on (A) c0372L,
(B) c(61331/21, (C) a1i33Y2L, and (D) a5133721, GABAA receptor isoforms
activity in the presence of 10
1.11\4 GABA. 100% represents full activation of the GABAA receptor. 50% to 20%
value represents
strong inhibition of GABAA receptor activity. Bumetanide (BTN) was used as a
negative control.
[0128] FIGURE 7A-B illustrate the inhibitory effect of select compounds at 10
11M concentration
on (A) oc4133721, and (B) a6P372L GABAA receptor isoforms activity in the
presence of 10 [tM GABA.
100% represents full activation of the GABAA receptor. 50% to 20% value
represents strong
inhibition of GABAA receptor activity. Bumetanide (BTN) was used as a negative
control.
[0129] FIGURE 8A-D illustrate the inhibitory effect of 10 11114 of select
compounds in the presence
of 10 1\4 GABA on (A) or.4133y2L, (B) 01.63372L, (C) al13372L, and (D)
a.513372L GABAA receptor
isoforms activity. 100% represents full activation of the GABAA receptor. 50%
to 20% value
represents strong inhibition of GABAA receptor activity. Bumetanide (BTN) was
used as a negative
control.
[0130] FIGURE 9A-J depicts the results from tail-flick assays.
[0131] FIGURE 10A-F depicts data on mIPSC frequency, mIPSC amplitude, mean
mIPSC decay
time, mean mIPSC rise time, and mean mIPSC half width for compound 3034.
[0132] FIGURE 11A-F depicts data on mIPSC frequency, mIPSC amplitude, mean
mIPSC decay
time, mean mIPSC rise time, and mean mIPSC half width for compound 6009.
[0133] FIGURE 12A-F depicts data on mIPSC frequency, mIPSC amplitude, mean
mIPSC decay
time, mean mIPSC rise time, and mean mIPSC half width for compound 7049.
Detailed Description
[0134] The foregoing and other aspects of the present invention will now be
described in more detail
with respect to embodiments described herein. It should be appreciated that
the invention can be
embodied in different forms and should not be construed as limited to the
embodiments set forth
herein. Rather, these embodiments are provided so that this disclosure will be
thorough and
complete, and will fully convey the scope of the invention to those skilled in
the art.
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Definitions
[0135] The terminology used in the description of the invention herein is for
the purpose of
describing particular embodiments only and is not intended to be limiting of
the invention. As used
in the description of the embodiments of the invention and the appended
claims, the singular forms
"a," "an" and "the" are intended to include the plural forms as well, unless
the context clearly
indicates otherwise. Also, as used herein, "and/or" refers to and encompasses
any and all possible
combinations of one or more of the associated listed items. Furthermore,
"about," as used herein
when referring to a measurable value such as an amount of a compound, dose,
time, temperature is
meant to encompass variations of 20%, 10%, 5%, 1%, 0.5%, or even 0.1% of the
specified amount.
Unless otherwise defined, all terms, including technical and scientific terms
used in the description,
have the same meaning as commonly understood by one of ordinary skill in the
art to which this
invention belongs.
[0136] "Administration" as used herein, refers broadly to any means by which a
composition is
given to a patient. A preferred route of administration is oral, and unless
otherwise indicated, any
reference herein to "administration" includes "oral administration."
[0137] "Alkenyl" as used herein, refers broadly to a straight or branched
chain hydrocarbon radical
having one or more double bonds and containing from 2 to 20 carbon atoms.
Examples of alkenyl
groups include propenyl, butenyl, pentenyl, and the like. "Cycloalkenyl" or
"cyclic alkenyl" as used
herein refers to carbocycles containing no heteroatoms, and includes mono-, bi-
, and tricyclic
saturated carbocycles, as well as fused rings systems. Examples of
cycloalkenyl groups include
cyclopropenyl, cyclopentenyl, cyclohexenyl, cyclopentadienyl, cyclohexadienyl,
and the like. Such
alkenyl and cycloalkenyl groups may be optionally substituted as described
herein.
[0138] "Alkyl" as used herein refers broadly to a straight or branched chain
saturated hydrocarbon
radical. "Alkyl" also refers broadly to cyclic (i.e., cycloalkyl) alkyl
groups. Examples of alkyl
groups include, but are not limited to, straight chained alkyl groups
including methyl, ethyl, n-
propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and branched alkyl
groups including isopropyl,
tert-butyl, iso-amyl, neopentyl, iso-amyl, and the like. "Cycloalkyl" or
"cyclic alkyl" as used herein
refers to carbocycles containing no heteroatoms, and includes mono-, bi- and
tricyclic saturated
carbocycles, as well as fused ring systems. Examples of cycloalkyl include
cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, and the like. The cycloalkyl can be
substituted or
unsubstituted, and cyclic alkyl groups including cyclopropyl, cyclopentyl,
cyclohexyl, cycloheptyl,
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and the like. Such alkyl groups may be optionally substituted as described
herein. When
substituted, the substituents include, but are not limited to, cycloalkyl;
hydroxy; alkoxy; aryl;
heteroaryl; amino optionally substituted by alkyl; carboxy; amido; carbamoyl
optionally substituted
by alkyl; aminosulfonyl optionally substituted by alkyl; alkylsulfonyl; acyl;
aroyl; heteroaroyl;
acyloxy; aroyloxy; heteroaroyloxy; alkoxycarbonyl; nitro; cyano; halogen;
perfluoroalkyl;and
heterocycloalkyl; with multiple degrees of substitution being allowed on the
alkyl group.
[0139] "Alkylcyano" refers broadly to a straight or branched chain, saturated
or partially unsaturated
hydrocarbon radical bonded to a cyano (i.e., group.
[0140] "Alkylhalo" refers broadly to a straight or branched chain, saturated
or partially unsaturated
hydrocarbon radical bonded to a halogen (e.g., fluoro, chloro, bromo, and
iodo).
[0141] "Alkaryl" or "arylalkyl" as used herein refers broadly to a straight or
branched chain,
saturated hydrocarbon radical bonded to an aryl group. Examples of alkaryl
groups include, but are
not limited to, benzyl, 4-chlorobenzyl, methylbenzyl, dimethylbenzyl,
ethylphenyl, propyl-(4-
nitrophenyl), and the like. Such alkaryl groups may be optionally substituted
described herein.
[0142] "Alkylene" as used herein refers broadly to a straight or branched
chain having two terminal
monovalent radical centers derived by the removal of one hydrogen atom from
each of the two
terminal carbon atoms of straight-chain parent alkane.
[0143] "Aryl" or "Ar" as used herein refers broadly to an optionally
substituted aromatic group or to
an optionally substituted aromatic group fused to one or more optionally
substituted aromatic
groups, optionally substituted with suitable substituents including, but not
limited to, alkyl; alkoxy;
alkylsulfanyl; alkylsulfenyl; alkylsulfonyl; oxo; hydroxy; mercapto; amino
optionally substituted by
alkyl; amido; carboxy; carbamoyl optionally substituted by alkyl;
aminosulfonyl optionally
substituted by alkyl; acyl; aroyl; heteroaroyl; acyloxy; aroyloxy;
heteroaroyloxy; alkoxycarbonyl;
nitro; cyano; halogen; or perfluoroalkyl; multiple degrees of substitution
being allowed. Examples
of aryl include, but are not limited to, phenyl, 2-naphthyl, 1-naphthyl, and
the like. In some
embodiments, two adjacent hydroxy groups on an aromatic group can form a
dioxolane.
[0144] "Alkoxy" as used herein alone or as part of another group, refers
broadly to an alkyl group,
as defined herein, appended to the parent molecular moiety through an oxy
group. In some
embodiments, the alkyl group can be interrupted by one or more heteroatoms
(e.g., 0, S, or N).
Examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy,
2-propoxy, butoxy,
tert-butoxy, pentyloxy, hexyloxy, ethyloxyethyl, and the like.
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[0145] "Alkaryloxy" or "oxyalkaryl" as used herein refers broadly to the group
-0-alkyl-aryl
wherein Ar is aryl. Examples include, but are not limited to, benzyloxy,
oxybenzyl, 2-naphthyloxy,
and oxy-2-naphthyl.
[0146] "Alkaryloxyalkyl" or "alkyloxyalkaryl" as used herein refers broadly to
the group
-alkyl-O-alkyl-aryl wherein Ar is aryl. Examples include, but are not limited
to benzyloxyethyl.
[0147] "Analogs," as used herein, refer broadly to the modification or
substitution of one or more
chemical moieties on a parent compound and may include derivatives, positional
isomers, and
prodrugs of the parent compound.
[0148] "Aryloxy" as used herein refers broadly to the group ¨Ar0 wherein Ar is
aryl or heteroaryl.
Examples include, but are not limited to, phenoxy, benzyloxy, and 2-
naphthyloxy.
[0149] "Amino" as used herein refers broadly to -NH2 in which one or both of
the hydrogen atoms
may optionally be replaced by alkyl, aryl or heteroaryl, where the alkyl,
aryl, and heteroaryl groups
is optionally substituted.
[0150] "Alkylthio" or "thioalkyl," as used herein alone or as part of another
group, refers broadly to
an alkyl group, as defined herein, appended to the parent molecular moiety
through a sulfur moiety.
Representative examples of alkylthio include, but are not limited to,
methylthio, thiomethyl,
ethylthio, thioethyl, n-propylthio, thio-n-propyl, isopropylthio, thio-
isopropyl, n-butylthio, thio-n-
butyl, and the like.
[0151] "Arylthio" or "thioaryl," as used herein refers broadly to the group
¨ArS wherein Ar is aryl.
Examples include, but are not limited to, phenylthio, thiophenyl, 2-
naphthylthio, and thio-2-
naphthyl.
[0152] "Alkarylthio" or "thioalkaryl" as used herein refers broadly to the
group -S-alkyl-aryl
wherein Ar is aryl. Examples include, but are not limited to, benzylthio,
thiobenzyl, 2-naphthylthio,
and thio-2-naphthyl.
[0153] "Alkylheterocycloalkyl" as used herein refers to as used herein refers
broadly to a straight or
branched chain, saturated hydrocarbon radical bonded to a heterocycloalkyl
group.
[0154] "Biocompatible polymer" as used herein refers broadly to a polymer
moiety that is
substantially non-toxic and does not tend to produce substantial immune
responses, clotting or other
undesirable effects. Accordingly to some embodiments of the present invention,
polyalkylene glycol
is a biocompatible polymer where, as used herein, polyalkylene glycol refers
to straight or branched
polyalkylene glycol polymers such as polyethylene glycol, polypropylene
glycol, and polybutylene
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glycol, and further includes the monoalkylether of the polyalkylene glycol. In
some embodiments of
the present invention, the polyalkylene glycol polymer is a lower alkyl
polyalkylene glycol moiety
such as a polyethylene glycol moiety (PEG), a polypropylene glycol moiety, or
a polybutylene
glycol moiety. PEG has the formula - HO(CH2CH20)H, where n can range from
about 1 to about
4000 or more. In some embodiments, n is 1 to 100, and in other embodiments, n
is 5 to 30. The
PEG moiety can be linear or branched. In further embodiments, PEG can be
attached to groups such
as hydroxyl, alkyl, aryl, acyl, or ester. In some embodiments, PEG can be an
alkoxy PEG, such as
methoxy-PEG (or mPEG), where one terminus are a relatively inert alkoxy group,
while the other
terminus are a hydroxyl group.
[0155] "Bioavailability", as used herein, refers broadly to the availability
of a drug to an animal
following administration and may be used interchangeably with "systemic
exposure" (e.g., the
bioavailability of a drug is expressed as the systemic exposure of a cell to
drugs).
[0156] "Carboxy" as used herein refers broadly to the group -CO2H.
[0157] "Cycloalkyl" as used herein refers to carbocycles containing no
heteroatoms, and includes
mono-, hi- and tricyclic saturated carbocycles, as well as fused ring systems.
Examples of
cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the
like. The cycloalkyl
can be substituted or unsubstituted.
[0158] "Effective amount" or "effective," as used herein, refers broadly to a
dose that causes a relief
of symptoms of a disease or disorder as noted through clinical testing and
evaluation, patient
observation, and/or the like. "Effective amount" or "effective" further can
further designate a dose
that causes a detectable change in biological or chemical activity. The
detectable changes may be
detected and/or further quantified by one skilled in the art for the relevant
mechanism or process.
Moreover, "effective amount" or "effective" can designate an amount that
maintains a desired
physiological state, i.e., reduces or prevents significant decline and/or
promotes improvement in the
condition of interest. As are generally understood in the art, the dosage will
vary depending on the
administration routes, symptoms, and body weight of the patient but also
depending upon the
compound being administered.
[0159] "Halo" as used herein refers broadly to bromo, chloro, fluor , or iodo.
Alternatively, the
term "halide" as used herein refers broadly to bromide, chloride, fluoride, or
iodide.
[0160] "Hydroxy" as used herein refers broadly to the group ¨OH.
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[0161] "Heteroaryl" as used herein refers to an aromatic five- or six-membered
ring where at least
one atom consists of a heteroatom (e.g., 0, S. or N), and the remaining atoms
are carbon. The five-
membered rings have two double bands, and the six-membered rings have three
double bonds. The
heteroaryl group can be monocyclic or bicyclic (fused or non-fused). Examples
of monocyclic
heteroaryl groups include furanyl, thiophene-yl, pyrrolyl, oxazolyl,
thiazolyl, imidazolyl, pyrazolyl,
isoxazolyl, isothiazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, pyridinyl,
pyridazinyl, pyrimidinyl,
pyrazinyl, triazinyl, and the like. Examples of bicyclic heteroaryl groups
include indolizinyl,
indolyl, isoindolyl, benzofuranyl, benzothiophene-yl, indazolyl,
benzimidazolyl, benzthiazolyl,
purinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl,
quinoxalinyl, napthyridinyl,
pteridinyl, and the like. The heteroaryl group can be substituted or
unsubstituted.
[0162] "Heterocycloalkyl" as used herein refers to a cycloalkyl group where at
least one of the
carbon atoms in the ring is replaced by a heteroatorn (e.g., 0, S. or N). The
heterocycloalkyl group
can be monocyclic or bicyclic (fused or non-fused). Examples of monocyclic
heterocycloalkyl
groups include azetidinyl, pyrrolidinyl, piperidinyl, homopiperidinyl,
piperazinyl, morpholinyl,
thiomorpholinyl, 1-oxothiomorpholinyl, 1,1-dioxothiomorpholinyl,
tetrahydrooxazolyl,
tetrahydroisoxazolyl, tetrahydroimidazolyl, tetrahydropyrazolyl,
tetrahydrothiazolidinyl,
tetrahydroisothiazolidinyl, tetrahydropyrimidinyl, tetrahydropyridazinyl, 4-
piperadonyl, and the like.
Examples of bicyclic non-fused heterocycloalkyl groups include quinuclidinyl,
adamantyl, 2-
azobicyclo[3.2.1]octyl, and the like. Examples of fused heterocycloalkyl
groups include any of the
aforementioned monocyclic heterocycloalkyl groups fused with another
cycloalkyl or
heterocycloalkyl group. Examples of non-fused heterocycloalkyl groups include
spirocycles of any
of the aforementioned monocyclic heterocycloalkyl groups with another
cycloalkyl or
heterocycloalkyl group. The heterocycloalkyl group can be substituted or
unsubstituted. When
substituted, the substituents include, but are not limited to, cycloalkyl;
hydroxy; alkoxy; aryl;
heteroaryl; amino optionally substituted by alkyl; carboxy; amido; carbamoyl
optionally substituted
by alkyl; aminosulfonyl optionally substituted by alkyl; alkylsulfonyl; acyl;
aroyl; heteroaroyl;
acyloxy; aroyloxy; heteroaroyloxy; alkoxycarbonyl; nitro; cyano; halogen;
perfluoroalkyl;and
heterocycloalkyl; with multiple degrees of substitution being allowed on the
alkyl group.
[0163] "Increased" or "increase" as used herein, refers broadly to a
quantified change in a
measurable quality that is larger than the margin of error inherent in the
measurement technique,
preferably an increase by about 2-fold or greater relative to a control
measurement, more preferably
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an increase by about 5-fold or greater, and most preferably an increase by
about 10-fold or greater.
In particular, the term "increase," as used herein, refers broadly to make
greater, as in number, size,
strength, or quality; add to; and/or augment. "Increase," as used herein, also
encompasses expand,
extend, prolong, augment, enlarge, grow, develop, and/or swell. "Increase," as
used herein,
additionally encompasses where a given parameter (e.g., level, amount, size,
scope, duration,
weight) is greater, as in number, size, strength, or quality, than it once
was. Furthermore, the
"increase" in any number, size, strength, or quality of a given parameter may
be determined as
between two or more time points, especially if before or after a treatment,
event, or administration of
an agent or composition. Further, "increase" refers broadly to significant or
detectable, functionally,
analytically, and/or clinically, changes in the number, size, strength, or
quality of a given parameter
in question.
[0164] "Mammal" as used herein, refers broadly to any and all warm-blooded
vertebrate animals of
the class Mammalia, including humans, characterized by a covering of hair on
the skin and, in the
female, milk-producing mammary glands for nourishing the young. Examples of
mammals include
but are not limited to alpacas, armadillos, capybaras, cats, chimpanzees,
chinchillas, cattle, dogs,
goats, gorillas, hamsters, horses, humans, lemurs, llamas, mice, non-human
primates, pigs, rats,
sheep, shrews, and tapirs. Mammals include but are not limited to bovine,
canine, equine, feline,
murine, ovine, porcine, primate, and rodent species. Mammal also includes any
and all those listed
on the Mammal Species of the World maintained by the National Museum of
Natural History,
Smithsonian Institution in Washington DC, hereby incorporated by reference in
its entirety.
[0165] "N-oxide" or "amine N-oxide" as used herein refers broadly to a
chemical structure having
an N-0 bond where the nitrogen is positively charged and the oxygen is
negatively charged.
[0166] "N-substituted sulfonamide" as used herein refers broadly to a chemical
structure having the
-S02-NH(R) group. In this context, the R-group includes, but is not limited to
lower alkyl (e.g., C1-
C5 alkyl), lower alkenyl (e.g., C2-C6 alkenyl), alkaryl, aryl, cycloalkenyl,
cycloalkyl,
dialkylaminoalkyl, heterocycloalkyl, and heteroaryl.
[0167] "N,N-disubstituted sulfonamide" as used herein refers broadly to a
chemical structure having
the -S02-NRRI group. In this context, the R and R' are the same or different
and are independently
lower alkyl, lower alkenyl, alkaryl, aryl, cycloalkenyl, cycloalkyl,
dialkylaminoalkyl,
heterocycloalkyl, heteroaryl or taken together with the nitrogen atom to which
they are attached
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form a 4-8 member cycle which can be substituted or unsubstituted and can have
one or more
heteroatoms (e.g., N, 0, or 5).
[01681 "Parasynaptic" as used herein, refers broadly to receptors (e.g., GABAA
receptors) located
outside or in the perimeter of the synapse (e.g., synaptic cleft). Also,
"parasynaptic" refers broadly
to any receptors located perisynaptically, extrasynaptically, and
presynaptically.
[0169] "Patient" as used herein, refers broadly to any animal who is in need
of treatment either to
alleviate a disease state or to prevent the occurrence or reoccurrence of a
disease state. Also,
"Patient" as used herein, refers broadly to any animal who has risk factors, a
history of disease,
susceptibility, symptoms, signs, was previously diagnosed, is at risk for, or
is a member of a patient
population for a disease. The patient may be a clinical patient such as a
human or a veterinary
patient such as a companion, domesticated, livestock, exotic, or zoo animal.
Animals may be
mammals, reptiles, birds, amphibians, or invertebrates.
[0170] A "pharmaceutical composition" refers to a chemical or biological
composition suitable for
administration to a subject (e.g., mammal). Such compositions may be
specifically formulated for
administration via one or more of a number of routes, including but not
limited to buccal, cutaneous,
epicutaneous, epidural, infusion, inhalation, intraarterial, intracardial,
intracerebroventricular,
intradermal, intramuscular, intranasal, intraocular, intraperitoneal,
intraspinal, intrathecal,
intravenous, oral, parenteral, pulmonary, rectally via an enema or
suppository, subcutaneous,
subdermal, sublingual, transdermal, and transmucosal. In addition,
administration can by means of
capsule, drops, foams, gel, gum, injection, liquid, patch, pill, porous pouch,
powder, tablet, or other
means of administration.
[0171] A "pharmaceutical excipient" or a "pharmaceutically acceptable
excipient" is a carrier,
usually a liquid, in which an active therapeutic agent is formulated. The
excipient generally does not
provide any pharmacological activity to the formulation, though it may provide
chemical and/or
biological stability, and release characteristics. Exemplary formulations can
be found, for example,
in Remington, The Science And Practice of Pharmacy (20th Ed.) (Gennaro, A. R.,
Chief Editor),
Philadelphia College of Pharmacy and Science (2000).
[0172] As used herein "pharmaceutically acceptable carrier" or "excipient"
includes any and all
solvents, dispersion media, coatings, antibacterial and antifungal agents,
isotonic and absorption
delaying agents that are physiologically compatible. In one embodiment, the
carrier is suitable for
parenteral administration. Alternatively, the carrier can be suitable for
intravenous, intraperitoneal,
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intramuscular, sublingual, or oral administration. Pharmaceutically acceptable
carriers include
sterile aqueous solutions or dispersions and sterile powders for the
extemporaneous preparation of
sterile injectable solutions or dispersion. The use of such media and agents
for pharmaceutically
active substances are well known in the art. Except insofar as any
conventional media or agent is
incompatible with the active compound, use thereof in the pharmaceutical
compositions of the
invention is contemplated. Supplementary active compounds can also be
incorporated into the
compositions.
[0173] "Pharmaceutically acceptable salt" as used herein, refers broadly to a
salt form of a
compound permitting its use or formulation as a pharmaceutical and which
retains the biological
effectiveness of the free acid and base of the specified compound and that is
not biologically or
otherwise undesirable.
[0174] "Prophylactically effective amount" as used herein, refers broadly to
the amount of a
compound that, when administered to a patient for prophylaxis of a disease or
prevention of the
reoccurrence of a disease, is sufficient to effect such prophylaxis for the
disease or reoccurrence.
The prophylactically effective amount may be an amount effective to prevent
the incidence of signs
and/or symptoms. The "prophylactically effective amount" may vary depending on
the disease and
its severity and the age, weight, medical history, predisposition to
conditions, preexisting conditions,
of the patient to be treated.
[0175] "Prophylaxis," as used herein, refers broadly to a course of therapy
where signs and/or
symptoms are not present in the patient, are in remission, or were previously
present in a patient.
Prophylaxis includes preventing disease occurring subsequent to treatment of a
disease in a patient.
Further, prevention includes treating patients who may potentially develop the
disease, especially
patients who are susceptible to the disease (e.g., members of a patient
population, those with risk
factors, or at risk for developing the disease).
[0176] "Protective," as used herein, refers broadly to reducing the incidence
or severity of the
disease in a patient. Protective, as used herein, refers broadly to inhibiting
the disease, arresting the
development of the disease or its clinical symptoms, and/or causing regression
of the disease or its
clinical symptoms. Prevention also preferably includes preventing or reducing
incidence or severity
of disease in a patient.
[0177] "Protective effect amount," as used herein, refers broadly to the
amount of a compound that,
when administered to a patient reduces the severity of the incidence of signs
and/or symptoms, slows
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the development of the incidence of signs and/or symptoms, prevents the
development of the
incidence of signs and/or symptoms. The "protective effective amount" may vary
depending on the
disease and its severity and the age, weight, medical history, predisposition
to conditions, preexisting
conditions, of the patient to be treated.
[0178] "Quaternary ammonium" as used herein refers broadly to a chemical
structure having four
bonds to the nitrogen with a positive charge on the nitrogen in the "onium"
state, i.e., "R4N+" or
"quaternary nitrogen," wherein R is an organic substituent such as alkyl or
aryl. The term
"quaternary ammonium salt" as used herein refers broadly to the association of
the quaternary
ammonium cation with an anion.
[0179] "Signs" of disease, as used herein, refers broadly to any abnormality
indicative of disease,
discoverable on examination of the patient; an objective indication of
disease, in contrast to a
symptom, which is a subjective indication of disease.
[0180] "Symptoms" of disease as used herein, refers broadly to any morbid
phenomenon or
departure from the normal in structure, function, or sensation, experienced by
the patient and
indicative of disease.
[0181] "Subjects" as used herein, refers broadly to anyone suitable to be
treated according to the
present invention include, but are not limited to, avian and mammalian
subjects, and are preferably
mammalian. Mammals of the present invention include, but are not limited to,
canines, felines,
bovines, caprines, equines, vines, porcines, rodents (e.g., rats and mice),
lagomorphs, primates,
humans, and the like, and mammals in utero. Any mammalian subject in need of
being treated
according to the present invention is suitable. Human subjects are preferred.
Human subjects of
both genders and at any stage of development (i.e., neonate, infant, juvenile,
adolescent, adult) can
be treated according to the present invention.
[0182] The present invention can also be carried out on animal subjects,
particularly mammalian
subjects such as mice, rats, dogs, cats, cattle, goats, sheep, and horses for
veterinary purposes, and
for drug screening and drug development purposes. The present invention can
also be carried out on
avians including chickens, ducks, turkeys, geese, quail, pheasant, ratites
(e.g., ostrich) and
domesticated birds (e.g., parrots and canaries), and birds in ovo. "Subjects"
is used interchangeably
with "patients."
[0183] "Solvate" as used herein refers broadly to a molecular complex of a
compound with one or
more solvent molecules in a stoichiometric or non-stoichiometric amount. Such
solvent molecules
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are those commonly used in the pharmaceutical arts, e.g., water, ethanol, and
the like. A molecular
complex of a compound or moiety of a compound and a solvent can be stabilized
by non-covalent
intra-molecular forces such as, electrostatic forces, van der Waals forces,
and hydrogen bonds. The
term "hydrate" refers to a complex in which the one or more solvent molecules
are water including
monohydrates and hemi-hydrates. Examples of solvates, without limitation,
include compounds of
the invention in combination with water, I-propanol, 2-propanol, ethanol,
methanol, DMSO, ethyl
acetate, acetic acid, or ethanolamine.
[0184] "Substituted" as used herein refers broadly to replacement of one or
more of the hydrogen
atoms of the group replaced by substituents known to those skilled in the art
and resulting in a stable
compound as described below. Examples of suitable replacement groups include,
but are not limited
to, alkyl, acyl, alkenyl, alkynyl cycloalkyl, aryl, alkaryl, hydroxy, thio,
alkoxy, aryloxy, acyl, amino,
amido, carboxy, carboxyalkyl, thiocarboxyalkyl, carboxyaryl, thiocarboxyaryl,
halo, oxo, mercapto,
sulfinyl, sulfonyl, sulfonamido, amidino, carbamoyl, cycloalkyl,
heterocycloalkyl,
dialkylaminoalkyl, carboxylic acid, carboxamido, haloalkyl, dihaloalkyl,
trihaloalkyl, trihaloalkoxy,
alkylthio, aralkyl, alkylsulfonyl, arylthio, amino, alkylami no, dialkylamino,
guanidino, ureido, nitro
and the like. Substitutions are permissible when such combinations result in
compounds stable for
the intended purpose. For example, substitutions are permissible when the
resultant compound is
sufficiently robust to survive isolation to a useful degree of purity from a
reaction mixture, and
formulation into a therapeutic or diagnostic agent or reagent.
[0185] "Therapy" or "therapeutic" as used herein refers broadly to treating a
disease, arresting or
reducing the development of the disease or its clinical symptoms, and/or
relieving the disease,
causing regression of the disease or its clinical symptoms. Therapy
encompasses prophylaxis,
prevention, treatment, cure, regimen, remedy, minimization, reduction,
alleviation, and/or providing
relief from a disease, signs, and/or symptoms of a disease. Therapy
encompasses an alleviation of
signs and/or symptoms in patients with ongoing disease signs and/or symptoms
(e.g., pain,
inflammation.) Therapy also encompasses "prophylaxis" and "prevention."
Prophylaxis includes
preventing disease occurring subsequent to treatment of a disease in a patient
or reducing the
incidence or severity of the disease in a patient. The term "reduced," for
purpose of therapy, refers
broadly to the clinical significant reduction in signs and/or symptoms.
Therapy includes treating
relapses or recurrent signs and/or symptoms (e.g., of pain.) Therapy
encompasses but is not limited
to precluding the appearance of signs and/or symptoms anytime as well as
reducing existing signs
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and/or symptoms and eliminating existing signs and/or symptoms. Therapy
includes treating
chronic disease ("maintenance") and acute disease.
[0186] Therapy can be for patients with risk factors, at risk patients in a
susceptible population,
patients with a history of disease, patients with symptoms, patients with
signs, patients with signs but
no symptoms, and patients with symptoms but no signs. Therapy can also be for
patients without
risk factors, not at risk, patients not in a susceptible population, patients
with no history of disease,
patients with no symptoms, patients without signs. Therapy can alleviate,
allay, abate, assuage,
curtail, decrease, ease, lessen, lighten, make better, make healthy, mitigate,
mollify, pacify, relieve,
rehabilitate, remedy, repair, and/or soothe a disease, disease signs, and/or
disease symptoms.
[0187] "Treating" or "treatment," as used herein, refers broadly to a course
of therapy where signs
and/or symptoms are present in the patient. The term "reduced," for purpose of
therapy, refers
broadly to clinically significant reduction in signs and/or symptoms.
Treatment includes treating
chronic disease ("maintenance") and acute disease. Treatment can be for
patients with risk factors,
at risk patients in a susceptible population, patients with a history of
disease, and/or patients with
symptoms, patients with signs. Treatment can alleviate, allay, abate, assuage,
curtail, decrease, ease,
lessen, lighten, make better, make healthy, mitigate, mollify, pacify,
relieve, rehabilitate, remedy,
repair, and/or soothe a disease, disease signs, and/or disease symptoms. By
the terms "treating" or
"treatment" of a disorder involving the Na.+K.+Cl- co-transporters, it is
intended that the severity of
the disorder or the symptoms of the disorder are reduced, or the disorder is
partially or entirely
eliminated, as compared to that which would occur in the absence of treatment.
Treatment does not
require the achievement of a complete cure of the disorder. By the terms
"preventing" or
"prevention" of the disorder involving the Nalc'Cl- co-transporters, it is
intended that the inventive
methods eliminate or reduce the incidence or onset of the disorder, as
compared to that which would
occur in the absence of treatment. Alternatively stated, the present methods
slow, delay, control, or
decrease the likelihood or probability of the disorder in the subject, as
compared to that which would
occur in the absence of treatment. Further, the terms "treating" or
"treatment" of a disorder
involving the GABAA receptor, are intended that the severity of the disorder
or the symptoms of the
disorder are reduced, or the disorder is partially or entirely eliminated, as
compared to that which
would occur in the absence of treatment. Treatment does not require the
achievement of a complete
cure of the disorder.
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Compounds
[0188] According to some embodiments, the present invention provides novel
compounds. Thus,
any of the R groups as defined herein can be excluded or modified in order to
exclude a known
compound and/or provide a novel compound. Also, any of the R groups as defined
herein can be
excluded from the compounds of the present invention, particularly with
reference to denoting novel
compounds of the present invention. Compounds of the present invention include
compounds
according to formula I, II, III or IV.
[0189] Embodiments of the present invention further provide intermediate
compounds formed
through the synthetic methods described herein to provide compounds of Formula
I-TV. The
intermediate compounds may possess utility as therapeutic agents for the range
of indications
described herein and/or reagents for further synthesis methods and reactions.
[0190] In some embodiments, the present invention encompasses the following
compounds,
including esters, amides, N-substituted sulfonamides and N,N-disubstituted
sulfonamides thereof.
Synthetic Methods
[0191] Embodiments of the present invention provide methods of modifying
compounds of the
present invention to increase their lipophilicity. The lipophilicity can be
measured by determining
the hydrophile-lipophile balance (HLB) or the partition coefficient (e.g., the
distribution of a
compound between water and octanol). In some embodiments, the compound is a
diuretic or
diuretic-like compound, and in particular embodiments, the compound is termed
a "loop diuretic."
For a discussion of pharmacological properties of diuretics. See generally,
Hardman, Limbird, and
Gilman, (Eds.) (2001) Goodman & Gilman's The Pharmacological Basis of
Therapeutics, McGraw-
Hill Medical Publishing Division (10th Ed.) Further included are compounds
that affect cation-
chloride cotransporters. Furosemide and bumetanide are classic examples of
NKCC antagonists.
[0192] Starting materials for synthesizing compounds of the present invention
can further include
compounds described in U.S. Patent No. 3,634,583; U.S. Patent No. 3,806,534;
U.S. Patent No.
3,058,882; U.S. Patent No. 4,010,273; U.S. Patent No. 3,665,002; and U.S.
Patent No. 3,665,002.
[0193] Compounds of the present invention can include isomers, tautomers,
zwitterions,
enantiomers, diastereomers, racemates, or stereochemical mixtures thereof.
Compounds of the
present invention can also comprise isosteres.
[0194] The term "isosteres" as used herein broadly refers to elements,
functional groups,
substituents, molecules, or ions having different molecular formulae but
exhibiting similar or
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identical physical properties. For example, tetrazole is an isostere of
carboxylic acid because it
mimics the properties of carboxylic acid even though they both have different
molecular formulae.
Typically, two isosteric molecules have similar or identical volumes and
shapes. Other physical
properties that isosteric compounds usually share include boiling point,
density, viscosity, and
thermal conductivity. However, certain properties are usually different:
dipolar moments, polarity,
polarization, size, and shape since the external orbitals may be hybridized
differently.
[01951 The term "isomers" as used herein refers broadly to compounds having
the same number and
kind of atoms, and hence the same molecular weight, but differing with respect
to the arrangement or
configuration of the atoms in space. Additionally, the term "isomers" includes
stereoisomers and
geometric isomers. The terms "stereoisomer" or "optical isomer" as used herein
refer to a stable
isomer that has at least one chiral atom or restricted rotation giving rise to
perpendicular
dissymmetric planes (e.g., certain biphenyls, allenes, and Spiro compounds)
and can rotate plane-
polarized light. Because asymmetric centers and other chemical structure can
exist in some of the
compounds of the present invention, which may give rise to stereoisomerism,
the invention
contemplates stereoisomers and mixtures thereof. The compounds of the present
invention and their
salts can include asymmetric carbon atoms and may therefore exist as single
stereoisomers,
racemates, and as mixtures of enantiomers and diastereomers. Typically, such
compounds will be
prepared as a racemic mixture. If desired, however, such compounds can be
prepared or isolated as
pure stereoisomers, i.e., as individual enantiomers or diastereomers, or as
stereoisomer-enriched
mixtures. Tautomers are readily interconvertible constitutional isomers and
there is a change in
connectivity of a ligand, as in the keto and enol forms of ethyl acetoacetate
(including tautomers of
any said compounds.) Zwitterions are inner salts or dipolar compounds
possessing acidic and basic
groups in the same molecule. At neutral pH, the cation and anion of most
zwitterions are equally
ionized.
Pharmaceutical Compositions
[01961 The compounds (e.g., analogs, derivatives, and prodrugs) of the present
invention or
pharmacologically acceptable salts thereof may be formulated into
pharmaceutical compositions of
various dosage forms. To prepare the pharmaceutical compositions of the
invention, at least one
compound, or pharmaceutically acceptable salts thereof, as the active
ingredient is intimately mixed
with appropriate carriers and additives according to techniques well known to
those skilled in the art
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of pharmaceutical formulations. Remington, The Science And Practice of
Pharmacy (20th Ed.)
(Gennaro, A. R., Chief Editor), Philadelphia College of Pharmacy and Science
(2000).
[0197] Pharmaceutically acceptable salts of the compounds described herein
include the salt form of
the compound permitting its use or formulation as a pharmaceutical and which
retains the biological
effectiveness of the free acid and base of the specified compound and that is
not biologically or
otherwise undesirable. Examples of such salts are described in Wermuth and
Stahl, (Eds.) (2002)
Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Wiley-Verlag
Helvetica Acta,
Zilrich, herein incorporated by references in its entirety. Examples of such
salts include alkali metal
salts and addition salts of free acids and bases. Examples of pharmaceutically
acceptable salts,
without limitation, include sulfates, pyrosulfates, bisulfates, sulfites,
bisulfites, phosphates,
monohydrogenphosphates, dihydrogen phosphates, metaphosphates, pyrophosphates,
chlorides,
bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates,
formates, isobutyrates,
caproates, heptanoates, propiolates, oxalates, malonates, succinates,
suberates, sebacates, fumarates,
maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methylbenzoates,
dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates,
xylenesulfonates,
phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, y-
hydroxybutyrates,
glycollates, tartrates, methanesulfonates, ethane sulfonates,
propanesulfonates, toluenesulfonates,
naphthalene-l-sulfonates, naphthalene-2-sulfonates, and mandelates. In some
embodiments,
pharmaceutically acceptable salt includes sodium, potassium, calcium,
ammonium,
trialkylarylammonium, and tetraalkylammonium salts.
[0198] Similarly, compositions for liquid preparations include solutions,
emulsions, dispersions,
suspensions, syrups, and elixirs, with suitable carriers and additives
including but not limited to
water, alcohols, oils, glycols, preservatives, flavoring agents, coloring
agents, and suspending
agents. Typical preparations for parenteral administration comprise the active
ingredient with a
carrier such as sterile water or parenterally acceptable oil including but not
limited to polyethylene
glycol, polyvinyl pyrrolidone, lecithin, arachis oil or sesame oil, with other
additives for aiding
solubility or preservation may also be included. In the case of a solution, it
can be lyophilized to a
powder and then reconstituted immediately prior to use. For dispersions and
suspensions,
appropriate carriers and additives include aqueous gums, celluloses,
silicates, or oils.
[0199] The pharmaceutical compositions according to embodiments of the present
invention include
those suitable for oral, rectal, topical, nasal, inhalation (e.g., via an
aerosol) buccal (e.g., sub-
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lingual), vaginal, topical (e.g., both skin and mucosal surfaces, including
airway surfaces),
transderrnal administration and parenteral (e.g., subcutaneous, intramuscular,
intradermal,
intraarticular, intrapleural, intraperitoneal, intrathecal, intracerebral,
intracranially, intraarterial, or
intravenous), although the most suitable route in any given case will depend
on the nature and
severity of the condition being treated and on the nature of the particular
active agent which is being
used. Pharmaceutical compositions of the present invention are particularly
suitable for oral,
sublingual, parenteral, implantation, nasal, and inhalational administration.
The carriers and
additives used for such pharmaceutical compositions can take a variety of
forms depending on the
anticipated mode of administration.
[0200] Compositions for oral administration may be solid preparations
including but not limited to
tablets, sugar-coated tablets, hard capsules, soft capsules, granules,
lozenges, and powders, with
suitable carriers and additives being starches, sugars, binders, diluents,
granulating agents,
lubricants, and disintegrating agents. Tablets and capsules represent
advantageous oral dosage forms
for many medical conditions because of their ease of use and higher patient
compliance.
[0201] Compositions for injection will include the active ingredient together
with suitable carriers
including organic solvents, propylene glycol-alcohol-water, isotonic water,
sterile water for injection
(USP), emulPhora)-alcohol-water, cremophor-EL or other suitable carriers
known to those skilled in
the art. These carriers may be used alone or in combination with other
conventional solubilizing
agents such as ethanol, a glycol, or other agents known to those skilled in
the art.
[0202] Where the compounds of the present invention are to be applied in the
form of solutions or
injections, the compounds may be used by dissolving or suspending in any
conventional diluent.
The diluents include but are not limited to physiological saline, Ringer's
solution, an aqueous
glucose solution, an aqueous dextrose solution, an alcohol, a fatty acid
ester, glycerol, a glycol, an
oil derived from plant or animal sources, and a paraffin. These preparations
may be prepared
according to any conventional method known to those skilled in the art.
[0203] Compositions for nasal administration may be formulated as aerosols,
drops, powders, and
gels. Aerosol formulations typically comprise a solution or fine suspension of
the active ingredient
in a physiologically acceptable aqueous or non-aqueous solvent. Such
formulations are typically
presented in single or multidose quantities in a sterile form in a sealed
container. The sealed
container can be a cartridge or refill for use with an atomizing device.
Alternatively, the sealed
container may be a unitary dispensing device such as a single use nasal
inhaler, pump atomizer or an
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aerosol dispenser fitted with a metering valve set to deliver an effective
amount, which is intended
for disposal once the contents have been completely used. When the dosage form
comprises an
aerosol dispenser, it will contain a propellant such as a compressed gas, air
as an example, or an
organic propellant including a fluorochlorohydrocarbon or fluorohydrocarbon.
[0204] Compositions suitable for buccal or sublingual administration include
but are not limited to
tablets, lozenges and pastilles, wherein the active ingredient is formulated
with a carrier such as
sugar and acacia, tragacanth or gelatin and glycerin.
[0205] Compositions for rectal administration include but are not limited to
suppositories containing
a conventional suppository base such as cocoa butter.
[0206] Compositions suitable for transdermal administration include but are
not limited to
ointments, gels, and patches.
[0207] The preferred forms of administration in the present invention are oral
forms known in the art
of pharmaceutics. The pharmaceutical compositions of the present invention may
be orally
administered as a capsule (hard or soft), tablet (film coated, enteric coated
or uncoated), powder or
granules (coated or uncoated) or liquid (solution or suspension). The
formulations may be
conveniently prepared by any of the methods well-known in the art. The
pharmaceutical
compositions of the present invention may include one or more suitable
production aids or excipients
including fillers, binders, disintegrants, lubricants, diluents, flow agents,
buffering agents,
moistening agents, preservatives, colorants, sweeteners, flavors, and
pharmaceutically compatible
carriers.
[0208] For each of the recited embodiments, the compounds can be administered
by a variety of
dosage forms as known in the art. Any biologically-acceptable dosage form
known to persons of
ordinary skill in the art, and combinations thereof, are contemplated.
Examples of such dosage
forms include, without limitation, chewable tablets, quick dissolve tablets,
effervescent tablets,
reconstitutable powders, elixirs, liquids, solutions, suspensions, emulsions,
tablets, multi-layer
tablets, bi-layer tablets, capsules, soft gelatin capsules, hard gelatin
capsules, caplets, lozenges,
chewable lozenges, beads, powders, gum, granules, particles, microparticles,
dispersible granules,
cachets, douches, suppositories, creams, topicals, inhalants, aerosol
inhalants, patches, particle
inhalants, implants, depot implants, ingestibles, injectables (including
subcutaneous, intramuscular,
intravenous, and intradermal), infusions, and combinations thereof.
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[0209] Other compositions known to those skilled in the art can also be
applied for percutaneous or
subcutaneous administration, such as plasters.
[0210] Further, in preparing such pharmaceutical compositions comprising the
active ingredient or
ingredients in admixture with components necessary for the formulation of the
compositions, other
conventional pharmacologically acceptable additives may be incorporated,
including but are not
limited to excipients, stabilizers, antiseptics, wetting agents, emulsifying
agents, lubricants,
sweetening agents, coloring agents, flavoring agents, isotonicity agents,
buffering agents, and
antioxidants. Additives include but are not limited to starch, sucrose,
fructose, dextrose, lactose,
glucose, mannitol, sorbitol, precipitated calcium carbonate, crystalline
cellulose, carboxymethyl
cellulose, dextrin, gelatin, acacia, EDTA, magnesium stearate, talc,
hydroxypropylmethylcellulose,
and sodium rnetabisulfite.
[0211] Compounds of the present invention may be used in conjunction with
delivery systems that
facilitate delivery of the agents to the central nervous system. For example,
various blood brain
barrier permeability enhancers may be used, if desired, to transiently and
reversibly increase the
permeability of the blood brain barrier to a treatment agent. Such BBB
permeability enhancers
include but are not limited to leukotrienes, bradykinin agonists, histamine,
tight junction disruptors
(e.g., zonulin, zot), hyperosmotic solutions (e.g., mannitol), cytoskeletal
contracting agents, and
short chain alkyl glycerols (e.g., 1-0-pentylglycerol). Oral, sublingual,
parenteral, implantation,
nasal and inhalational routes can provide delivery of the active agent to the
CNS. In some
embodiments, the compounds of the present invention may be administered to the
CNS with
minimal effects on the peripheral nervous system.
[0212] Pharmaceutical compositions typically must be sterile and stable under
the conditions of
manufacture and storage. The composition can be formulated as a solution,
microemulsion,
liposome, or other ordered structure suitable to high drug concentration. The
carrier can be a solvent
or dispersion medium containing, for example, water, ethanol, polyol (e.g.,
glycerol, propylene
glycol, and liquid polyethylene glycol), and suitable mixtures thereof. The
proper fluidity can be
maintained, for example, by the use of a coating such as lecithin, by the
maintenance of the required
particle size in the case of dispersion and by the use of surfactants.
[0213] In many cases, it will be preferable to include isotonic agents, for
example, sugars,
polyalcohols such as mannitol, sorbitol, or sodium chloride in the
composition. Prolonged
absorption of the injectable compositions can be brought about by including in
the composition an
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agent which delays absorption, for example, monostearate salts and gelatin.
Moreover, the
compounds described herein can be formulated in a time release formulation,
for example in a
composition that includes a slow release polymer. The active compounds can be
prepared with
carriers that will protect the compound against rapid release, such as a
controlled release
formulation, including implants and microencapsulated delivery systems.
Biodegradable,
biocompatible polymers may be used, such as ethylene vinyl acetate,
polyanhydrides, polyglycolic
acid, collagen, polyorthoesters, polylactic acid and polylactic, polyglycolic
copolymers (PLO).
Many methods for the preparation of such formulations are known to those
skilled in the art.
[0214] Other compounds which can be included by admixture are, for example,
medically inert
ingredients (e.g., solid and liquid diluent), such as lactose,
dextrosesaccharose, cellulose, starch or
calcium phosphate for tablets or capsules, olive oil or ethyl oleate for soft
capsules and water or
vegetable oil for suspensions or emulsions; lubricating agents such as silica,
talc, stearic acid,
magnesium or calcium stearate and/or polyethylene glycols; gelling agents such
as colloidal clays;
thickening agents such as gum tragacanth or sodium alginate, binding agents
such as starches, arable
gums, gelatin, methylcellulose, carboxymethylcellulose or
polyvirkylpyrrolidone; disintegrating
agents such as starch, alginic acid, alginates or sodium starch glycolate;
effervescing mixtures;
dyestuff; sweeteners; wetting agents such as lecithin, polysorbates or
laurylsulphates; and other
therapeutically acceptable accessory ingredients, such as humectants,
preservatives, buffers and
antioxidants, which are known additives for such formulations.
[0215] In further embodiments, the present invention provides kits including
one or more containers
comprising pharmaceutical dosage units comprising an effective amount of one
or more compounds
of the present invention. Kits may include instructions, directions, labels,
marketing information,
warnings, or information pamphlets.
Prodrugs
[0216] The blood-brain barrier (BBB) is a physical barrier and system of
cellular transport
mechanisms between the blood vessels in the central nervous system (CNS) and
most areas of the
CNS itself. The BBB maintains homeostasis by restricting the entry of
potentially harmful
chemicals from the blood, and by allowing the entry of essential nutrients.
However, the BBB can
pose a formidable barrier to delivery of pharmacological agents to the CNS for
treatment of
disorders or maintaining or enhancing normal and desirable brain functions,
such as cognition,
learning, and memory. Prodrugs of the present invention are capable of passage
across the blood-
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brain barrier and may undergo hydrolysis by CNS esterases to provide the
active compound.
Further, the prodrugs provided herein may also exhibit improved
bioavailability, improved aqueous
solubility, improved passive intestinal absorption, improved transporter-
mediated intestinal
absorption, protection against accelerated metabolism, tissue-selective
delivery, less (or fewer) side
effects, lessened or no deleterious drug interaction with other medications,
and/or passive
enrichment in the target tissue.
[0217] The term "prodrug" is intended to refer to a compound that is converted
under physiological
conditions, by solvolysis or metabolically to a specified compound that is
pharmaceutically/pharmacologically active. The "prodrug" can be a compound of
the present
invention that has been chemically derivatized such that it retains some, all
or none of the bioactivity
of its parent drug compound and is metabolized in a subject to yield the
parent drug compound. The
prodrug of the present invention may also be a "partial prodrug" in that the
compound has been
chemically derivatized such that it retains some, all or none of the
bioactivity of its parent drug
compound and is metabolized in a subject to yield a biologically active
derivative of the compound.
[0218] Moreover, as shown in the previously presented schemes, prodrugs can be
formed by
attachment of biocompatible polymers, such as those previously described
including polyethylene
glycol (PEG), to compounds of the present invention using linkages degradable
under physiological
conditions. See also Schacht, et al. (1997) Poly(ethylene glycol) Chemistry
and Biological
Applications, American Chemical Society, San Francisco, CA 297-315. Attachment
of PEG to
proteins can be employed to reduce irnmunogenicity and/or extend the half-life
of the compounds
provided herein. Any conventional PEGylation method can be employed, provided
that the
PEGylated agent retains at least some pharmaceutical activity.
[0219] The present invention further provides prodrugs comprising the
compounds described herein.
The prodrugs can be formed utilizing a hydrolyzable coupling to the compounds
described herein.
Ettmayer, et al. (2004) J. Med. Chem. 47(10): 2394 ¨ 2404 Testa and Mayer
(2003) Hydrolysis in
Drug and Prodrug Metabolism: Chemistry, Biochemistry and Enzymology Wiley-
Verlag Helvetica
Chimica Acta, Zuerich (Chapters 1-12): 1-780.
Dosages
[0220] The amount of active compound in a therapeutic composition according to
this invention
may vary according to factors such as the disease state, age, gender, weight,
patient history, risk
factors, predisposition to disease, administration route, pre-existing
treatment regime (e.g., possible
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interactions with other medications), and weight of the individual. Dosage
regimens may be
adjusted to provide the optimum therapeutic response. For example, a single
bolus may be
administered, several divided doses may be administered over time, or the dose
may be
proportionally reduced or increased as indicated by the exigencies of
therapeutic situation.
[0221] It is especially advantageous to formulate parenteral compositions in
dosage unit form for
ease of administration and uniformity of dosage. Dosage unit form as used
herein refers to
physically discrete units suited as unitary dosages for the mammalian subjects
to be treated; each
unit containing a predetermined quantity of active compound calculated to
produce the desired
therapeutic effect in association with the required pharmaceutical carrier.
The specification for the
dosage unit forms of the invention are dictated by and directly dependent on
the unique
characteristics of the active compound and the particular therapeutic effect
to be achieved, and the
limitations inherent in the art of compounding such an active compound for the
treatment of
sensitivity in individuals. In therapeutic use for treatment of conditions in
mammals (e.g., humans)
for which the compounds of the present invention or an appropriate
pharmaceutical composition
thereof are effective, the compounds of the present invention may be
administered in an effective
amount. The dosages as suitable for this invention may be a composition, a
pharmaceutical
composition or any other compositions described herein.
[0222] For each of the recited embodiments, the dose for a patient can be
about 0.01, 0.02, 0.03,
0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.10 pg, as well as about 0.1, 0.2,
0.3, 0.4, 0.5, 0.6,0.7, 0.8, 0.9,
or 1.0 g, as well as about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 jig, as well as
about 10,11, 12, 13, 14, 15,
16, 17, 18, 19, or 20 pg, as well as about 10, 20, 25, 30, 35, 40, 45, 50, 55,
60, 65, 70, 75, 80, 85, 90,
95, or 100 gg, as well as about 100, 200, 300, 400, 500, 600, 700, 800, 900,
or 1,000 pg and all
increments therein. Preferably, the dose for a patient can be about 0.05-5 ptg
and all increments
therein. Alternatively, the dose for a patient can be about 1-10 pg and all
increments therein. The
dose for a patient can also be about 10-40 pg and all increments therein,
about 6-24 pg and all
increments therein, about 20-80 pg and all increments therein, about 40-80 jig
and all increments
therein, about 100-250 jig and all increments therein, or about 100-500 jig
and all increments
therein. More preferably, the dosage can be about 0.5, 1, 2, 5, 6, 10, 12, 18,
20, 24, 30, 40, 50, 80, or
90 pg. Preferably, the dosage can be 0.5, 2, 6, 8, 10, 12, 18, 20, 30, 40, or
80 pg.
[02231 Alternatively, for each of the recited embodiments, the dose for a
patient may be about 0.01,
0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.10 mg, as well as about
0.1, 0.2, 0.3, 0.4, 0.5, 0.6,
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0.7, 0.8, 0.9, or 1.0 mg, as well as about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10
mg, as well as about 10, 11, 12,
13, 14, 15, 16, 17, 18, 19, or 20 mg, as well as about 10, 20, 25, 30, 35, 40,
45, 50, 55, 60, 65, 70, 75,
80, 85, 90, 95 or 100 mg, as well as about 100, 200, 300, 400, 500, 600, 700,
800, 900, or 1,000 mg
and all increments therein. Preferably, the dose for a patient may be about
0.05-5 mg and all
increments therein. Alternatively, the dose for a patient may be about 1-10 mg
and all increments
therein. The dose for a patient may also be about 10-40 mg and all increments
therein, about 6-
24 mg and all increments therein, about 20-80 mg and all increments therein,
about 40-80 mg and
all increments therein, about 100-250 mg and all increments therein, or about
100-500 mg and all
increments therein. More preferably, the dosage may be about 0.5, 1, 2, 5, 6,
10, 12, 18, 20, 24, 30,
40, 50, 80, or 90 mg. Preferably, the dosage may be 0.5, 2, 6, 8, 10, 12, 18
20, 30, 40, or 80 mg.
[02241 Alternatively, for each of the recited embodiments, the dose for a
patient may be about 0.01,
0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.10 g, as well as about
0.1, 0.2, 0.3, 0.4, 0.5, 0.6,
0.7, 0.8, 0.9, or 1.0 g, as well as about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 g,
as well as about 10, 11, 12, 13,
14, 15, 16, 17, 18, 19, or 20 g, as well as about 10, 20, 25, 30, 35, 40,45,
50, 55, 60, 65, 70, 75, 80,
85, 90, 95 or 100 g, as well as about 100, 200, 300, 400, 500, 600, 700, 800,
900, or 1,000 g and all
increments therein. Preferably, the dose for a patient may be about 0.05-5 g
and all increments
therein. Alternatively, the dose for a patient may be about 1-10 g and all
increments therein. The
dose for a patient may also be about 10-40 g and all increments therein, about
6-24 g and all
increments therein, about 20-80 g and all increments therein, about 40-80 g
and all increments
therein, about 100-250 g and all increments therein, or about 100-500 g and
all increments therein.
More preferably, the dosage may be about 0.5, 1, 2, 5, 6, 10, 12, 18, 20, 24,
30, 40, 50, 80, or 90 g.
Preferably, the dosage may be 0.5, 2, 6, 8, 10, 12, 18 20, 30, 40, or 80 g.
[0225] For each of the recited embodiments, the dose for a patient can be
about 0.01, 0.02, 0.03,
0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.10 kg/kg, as well as about 0.1, 0.2,
0.3, 0.4, 0.5, 0.6, 0.7, 0.8,
0.9, or 1.0 kg/kg, as well as about 1, 2, 3,4, 5, 6, 7, 8, 9, or 10 kg/kg, as
well as about 10, 11, 12, 13,
14, 15, 16, 17, 18, 19, or 20 kg/kg, as well as about 10, 20,25, 30, 35, 40,
45, 50, 55, 60, 65, 70, 75,
80, 85, 90, 95 or 100 kg/kg, as well as about 100, 200, 300, 400, 500, 600,
700, 800, 900, or
1,000 kg/kg and all increments therein. Preferably, the dose for a patient can
be about 0.05-5 kg/kg
and all increments therein. Alternatively, the dose for a patient can be about
1-10 kg/kg and all
increments therein. The dose for a patient can also be about 10-40 kg/kg and
all increments therein,
about 6-24 kg/kg and all increments therein, about 20-80 kg/kg and all
increments therein, about
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40-80 jig/kg and all increments therein, about 100-250 jig/kg and all
increments therein, or about
100-500 jig/kg and all increments therein. More preferably, the dosage can be
about 0.5, 1, 2, 5, 6,
10, 12, 18, 20, 24, 30, 40, 50, 80, or 90 g/kg.
[0226] Preferably, the dosage can be 0.5, 2, 6, 8, 10, 12, 18, 20, 30, 40, or
80 jig/kg.
Alternatively, for each of the recited embodiments, the dose for a patient may
be about 0.01, 0.02,
0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.10 mg/kg, as well as about 0.1,
0.2, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, or 1.0 mg/kg, as well as about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10
mg/kg, as well as about 10,11,
12, 13, 14, 15, 16, 17, 18, 19, or 20 mg/kg, as well as about 10, 20, 25, 30,
35, 40,45, 50, 55, 60, 65,
70, 75, 80, 85, 90, 95 or 100 mg/kg, as well as about 100, 200, 300, 400, 500,
600, 700, 800, 900, or
1,000 mg/kg and all increments therein. Preferably, the dose for a patient may
be about 0.05-5
mg/kg and all increments therein. Alternatively, the dose for a patient may be
about 1-10 mg/kg and
all increments therein. The dose for a patient may also be about 10-40 mg/kg
and all increments
therein, about 6-24 mg/kg and all increments therein, about 20-80 mg/kg and
all increments therein,
about 40-80 mg/kg and all increments therein, about 100-250 mg/kg and all
increments therein, or
about 100-500 mg/kg and all increments therein. More preferably, the dosage
may be about 0.5, 1,
2, 5, 6, 10, 12, 18, 20, 24, 30, 40, 50, 80, 90, or 100 mg/kg. Preferably, the
dosage may be 0.5, 2, 6,
8, 10, 12, 18, 20, 30, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 80, 85, 90,
or 100 mg/kg.
[0227] Alternatively, for each of the recited embodiments, the dose for a
patient can be about 0.01,
0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.10 g/kg, as well as about
0.1, 0.2, 0.3, 0.4, 0.5, 0.6,
0.7, 0.8, 0.9, or 1.0 g/kg, as well as about 1, 2, 3,4, 5, 6, 7, 8, 9, or 10
g/kg, as well as about 10, 11,
12, 13, 14, 15, 16, 17, 18, 19, or 20 g/kg, as well as about 10, 20, 25, 30,
35, 40, 45, 50, 55, 60, 65,
70, 75, 80, 85, 90, 95 or 100 g/kg, as well as about 100, 200, 300, 400, 500,
600, 700, 800, 900, or
1,000 g/kg and all increments therein. Preferably, the dose for a patient can
be about 0.05-5 g/kg
and all increments therein. Alternatively, the dose for a patient can be about
1-10 g/kg and all
increments therein. The dose for a patient can also be about 10-40 g/kg and
all increments therein,
about 6-24 g/kg and all increments therein, about 20-80 g/kg and all
increments therein, about 40-
80 g/kg and all increments therein, about 100-250 g/kg and all increments
therein, or about 100-500
g/kg and all increments therein. More preferably, the dosage can be about 0.5,
1, 2, 5, 6, 10, 12, 18,
20, 24, 30, 40, 50, 80, or 90 g/kg. Preferably, the dosage can be 0.5, 2, 6,
8, 10, 12, 18 20, 30, 40, or
80 g/kg.
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[0228] For each of the recited embodiments, the dosage is typically
administered once, twice, or
thrice a day, although more frequent dosing intervals are possible. The dosage
may be administered
every day, every 2 days, every 3 days, every 4 days, every 5 days, every 6
days, and/or every 7 days
(once a week). In one embodiment, the dosage may be administered daily for up
to and including 30
days, preferably between 7-10 days. In another embodiment, the dosage may be
administered twice
a day for 10 days. If the patient requires treatment for a chronic disease or
condition, the dosage
may be administered for as long as signs and/or symptoms persist. The patient
may require
"maintenance treatment" where the patient is receiving dosages every day for
months, years, or the
remainder of their lives. In addition, the composition of this invention may
be to effect prophylaxis
of recurring symptoms. For example, the dosage may be administered once or
twice a day to
prevent the onset of symptoms in patients at risk, especially for asymptomatic
patients.
[0229] For each of the recited embodiments, the patient can receive
"pretreatment" with the
compounds described herein wherein the compounds described herein are
administered every day,
every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, and/or
every 7 days (once a
week). In one embodiment, the dosage can be administered daily for up to and
including 30 days,
preferably between 7-10 days. In another embodiment, the dosage can be
administered twice a day
for 10 days. If the patient requires treatment for a chronic disease or
condition requiring prolonged
treatment, the dosage of alkaline may be administered for as long as symptoms
persist.
[0230] In one embodiment, the compounds described herein are administered in
an initial dose of
20-80 mg on the first day of treatment and then at least two dosages of about
40 mg on the second
day. In another embodiment the compounds described herein are administered in
an initial dose of
0.5-2 mg on the first day of treatment and then at least two dosages of about
2 mg on the second
day. In another embodiment, the compounds described herein are administered in
an initial dose of
10-20 mg on the first day of treatment and then at least two dosages of about
20 mg on the second
day. In yet another embodiment, the compounds described herein are
administered in an initial dose
of 5-10 mg on the first day of treatment and then at least two dosages of
about 10 mg on the second
day.
[0231] For administration via injection, in one embodiment the treatment
begins as a course of 4
injections at 0, 12, 24, and 36 hours. The injections then may continue once,
twice, or thrice a day
for as long as signs and/or symptoms persists. Alternatively, the injections
may be maintained to
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prevent the recurrence of disease. Also, the injections may be administered as
a prophylaxis for
patients at risk, especially asymptomatic patients.
[0232] The dosage may be administered as a single dose, a double dose, a
triple dose, a quadruple
dose, and/or a quintuple dose. The dosages may be administered singularly,
simultaneously, and
sequentially.
[0233] For each of the recited embodiments, the dosage of the compounds
described herein may be
an effective amount of the compounds described herein, an amount effective for
prophylaxis, and for
acute treatment, or an amount effective for prevention. The dosage of the
compounds described
herein may be an amount of the compounds described herein effective to reduce
signs or symptoms
of a disease, an amount effective to prevent signs and/or symptoms of a
disease, to reduce the
severity of signs and/or symptoms of a disease, to eliminate signs and/or
symptoms of a disease, to
slow the development of signs or symptoms of a disease, to prevent the
development of signs and/or
symptoms of a disease, or effect prophylaxis of signs or symptoms of a
disease.
[0234] The dosage form may be any form of release known to persons of ordinary
skill in the art.
The compositions of the present invention may be formulated to provide
immediate release of the
active ingredient or sustained or controlled release of the active ingredient.
In a sustained release or
controlled release preparation, release of the active ingredient may occur at
a rate such that blood
levels are maintained within a therapeutic range but below toxic levels over
an extended period of
time (e.g., 4 to 24 hours). The preferred dosage forms include immediate
release, extended release,
pulse release, variable release, controlled release, timed release, sustained
release, delayed release,
long acting, and combinations thereof. The ability to obtain immediate
release, extended release,
pulse release, variable release, controlled release, timed release, sustained
release, delayed release,
long acting characteristics, and combinations thereof is known in the art.
[0235] It will be appreciated that the pharmacological activity of the
compositions may be
monitored using standard pharmacological models that are known in the art.
Furthermore, it will be
appreciated that the inventive compositions may be incorporated or
encapsulated in a suitable
polymer matrix or membrane for site-specific delivery, or may be
functionalized with specific
targeting agents capable of effecting site specific delivery. For instance,
the dosage form may be
made such that it preferably releases in the central nervous system or
peripheral nervous system.
These techniques, as well as other drug delivery techniques are well known in
the art. Determination
of optimal dosages for a particular situation is within the capabilities of
those skilled in the art. See
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e.g., Gennaro (2000) Remington, The Science And Practice of Pharmacy (20th
Ed.) Philadelphia
College of Pharmacy and Science.
[0236] In further embodiments, compounds according to the present invention
may be administered
1.5 to 6 mg daily, for example, 1 tablet or capsule three times a day. In some
embodiments,
compounds according to the present invention may be administered 60 to 240
mg/day, for example,
1 tablet or capsule three times a day. In other embodiments, compounds
according to the present
invention may be administered 10 to 20 mg daily, for, example, 1 tablet or
capsule once a day. In
some embodiments, compounds according to the present invention may be
administered 60 mg per
day. In other embodiments, compounds according to the present invention may be
administered 10
to 20 mg daily, for example, 1 tablet or capsule once a day. It should be
noted that lower doses may
be administered, particularly for intravenous administration. Moreover,
administration of a lower
dose than administered for the parent compound may prevent undesirable
peripheral effects such as
diuresis.
[0237] In additional further embodiments, compounds are administered at about
0.5, 1.0, or 2.0 mg;
compounds are administered at about 20-80 mg or two 40 mg doses daily;
compounds are
administered 0, 200, 500, or 1,250 mg/kg, preferably at about 10-30 mg/kg or
200-500 mg/kg;
compounds are administered at 5, 10, 20, 40, or 200 mg. More preferably, the
compounds are
administered orally and daily at about 1, 10, or 20 mg.
Routes of Administration
[0238] The compositions described herein may be administered in any of the
following routes:
buccal, epieutaneous, epidural, infusion, inhalation, intraarterial,
intracardial,
intracerebroventricular, intradermal, intramuscular, intranasal, intraocular,
intraperitoneal,
intraspinal, intrathecal, intravenous, oral, parenteral, pulmonary, rectally
via an enema or
suppository, subcutaneous, subdermal, sublingual, transdermal, and
transmucosal. The preferred
routes of administration are buccal and oral. The administration can be local,
where the composition
is administered directly, close to, in the locality, near, at, about, or in
the vicinity of, the site(s) of
disease, e.g., inflammation, or systemic, wherein the composition is given to
the patient and passes
through the body widely, thereby reaching the site(s) of disease. Local
administration can be
administration to the cell, tissue, organ, and/or organ system, which
encompasses and/or is affected
by the disease, and/or where the disease signs and/or symptoms are active or
are likely to occur.
Administration can be topical with a local effect, composition is applied
directly where its action is
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desired. Administration can be enteral wherein the desired effect is systemic
(non-local),
composition is given via the digestive tract. Administration can be
parenteral, where the desired
effect is systemic, composition is given by other routes than the digestive
tract.
Nutritional Compositions
[0239] The compositions of the compounds described herein may be used in (or
consumed in)
nutritional supplements; dietary supplements; medical foods; nutriceuticals;
food-stuffs such as
pharmaceutical-benefit foods (e.g., "phoods"); beverages including fortified
(e.g., orange juice with
calcium); traditional (e.g., regular oatmeal, whole-grain breads), and
"designer" products (e.g.,
protein bars, smart spreads, smart bars, energy bars). The compounds described
herein may be
formulated in health bars, confections, animal feeds, cereals, dietary
supplements, yogurts, cereal
coatings, foods, nutritive foods, functional foods, and combinations thereof.
Second Agents
[0240] Second agents for treatment in combination with compositions of the
present invention
include, but are not limited to, phenytoin, carbamazepine, barbiturates,
phenobarbital,
mephobarbital, trimethadione, mephenytoin, paramethadione, phenthenylate,
phenacemide,
metharbital, benzchlorpropamide, phensuximide, primi done, methsuximide,
ethotoin,
aminoglutethinide, diazepam, clonazepam, clorazepate, fosphenytoin,
ethosuximide, valproate,
felbamate, gabapentin, lacosamide, lamotrigine, retigabine, rufinamide,
topiramate, vigrabatrin,
pregabalin, tiagabine, zonisamide, clobazam, thiopental, midazolam, propofol,
levetiracetam,
oxcarbazepine, CCPene, GYK152466, serotonin receptor agonists, ergotamine,
dihydroergotamine,
sumatriptan, propranolol, metoprolol, atenolol, timolol, nadolol, nifeddipine,
nimodipine, verapamil,
aspirin, ketoprofen, tofenamic acid, mefenamic acid, naproxen, methysergide,
paracetamol,
clonidine, lisuride, iprazochrome, butalbital, benzodiazepines, divalproex
sodium and other similar
classes of compounds. See U.S. Patent No. 6,495,601 and U.S. Patent
Application Publication No.
2002/0082252.
[0241] For example, in addition to the composition described herein patients
may also be treated
with antidepressants (e.g., tricyclic antidepressants [e.g., amitriptyline
(Elavil ), desipramine
(Norpramin ), imipramine (Tofranil ), nortriptyline (Aventyl , Pamelor )1 ;
Serotonin and
norepinephrine reuptake inhibitors (SNRIs) [e.g., venlafaxine (Effexor ),
duloxetine (Cymbalta )];
norepinephrine and dopamine reuptake inhibitors (NDRIs) [e.g., bupropion
(Wellbutrin )];
combined reuptake inhibitors and receptor blockers [e.g., trazodone
(Desyre10), nefazodone
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(Serzone0), maprotiline, mirtazpine (Remeron )]; monamine oxidase inhibitors
(MAOIs) [e.g.,
isocarboxazid (Marplan0), phenelzine (Nardi10), tranlcypromine (Parnate )] and
selective
serotonin reuptake inhibitors (SSRIs) [e.g., citalopram (Celexa0),
escitalopram (Lexapro0),
fluoxetine (Prozac0), paroxetine (Paxi10, Pexeva0), sertraline (Zoloft0)]
fluvoxamine (Luvox0),
and amitriptyline); anticonvulsants to stabilize abnormal electrical activity
in the nervous system
caused by injured nerves (e.g., gabapentin (NEURONTIN0), pregabalin (LYRICA0),
carbamazepine (Tegreto10), lamotrigine (Lamictal0), topiramate (Topamax0),
felbamate
(Felbato10), tiagabine (Gabitri10), diazepam rectal (Diastat0), phenobarbital,
phenytoin (Dilantin )
primidone (Mysoline0), valproate (Depakote0), vigabatrin, oxcarbazepine
(Trileptal0), zonisamide
(Zonegran ), and levetiracetam (Keppra0)); steroids (e.g., corticosteroid);
analgesics (e.g.,
acetaminophen (Tylenol ), codeine (Tylenol #2,3,40), propoyl APA (Darvocet0),
propoeylphene
(Darvon0), fentanyl patch (duragesic0), hydromorphone (Palladone0), morphine
(MS Contin0),
oxycodone (Percocet0, OxyContine, Percodani0), pentazocine (Talwin MOO),
tramadol APAP
(Ultracet0), tramadol (Ultram0), hydrocodone APAP (Vicodin0)); lithium, and
non-steroidal anti-
inflammatory drugs (NSAID) (e.g., Tylenol , Motrin0, salicylates (e.g.,
acetylsalicylic acid
(Aspirin), amoxiprin, benorylate/benorilate, choline magnesium salicylate,
Diflunisal ,
ethenzamide, faislamine, methyl salicylate, magnesium salicylate, salicyl
salicylate, and
salicylamide), arylalkanoic acids (e.g., Diclofenac , Aceclofenac0,
Acemethacin , Alclofenae ,
Bromfenac , Etodolac , Indomethacin , Nabumetone0, Oxametacin0, Proglumetacin
,
Sulindac0, and Tolmetin0) 2-Arylpropionic acids (profens) (e.g., Ibuprofen ,
Alminoprofen ,
Carprofera, Dexibuprofen , Dexketoprofen , Fenbufen , Fenoprofen ,
Flunoxaprofen0,
Flurbiprofen , Ibuproxam , Indoprofen , Ketorolac , Loxoprofen0, Naproxen ,
Oxaprozin ,
Pirprofen , Suprofen0, Tiaprofenic acid); N-Arylanthranilic acids (fenamic
acids) (e.g., mefenamic
acid, flufenamic acid, meclofenamic acid, tolfenamic acid, pyrazolidine
derivatives,
Phenylbutazone , Ampyrone0, Azapropazone0, Clofezone , Kebuzone0, Metamizole ,
Mofebutazone , Oxyphenbutazone , Phenazone , and Sulfinpyrazone0); and oxicams
(e.g.,
Piroxicam0, Droxicam , Lornoxicam0, Meloxicam0, and Tenoxicam0).
[0242] Such second agents can be administered in the same formulation (e.g.,
the same pill) or in a
separate formulation as the compounds of the present invention. It is
preferred that the second
agents described above be co-administered with the compounds of the present
invention.
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The second agents described herein can be administered with the compounds of
the present
invention simultaneously, sequentially, prior to, or after administering of
the compounds of the
present invention. Where the administration of the second agents described
herein is simultaneous,
the second agent and the compounds of the present invention are administered
together or within a
very short time interval (e.g., 5 minutes). Where the administration of the
second agent is
administered as pre-treatment, the second agent is administered prior to
administration of the
compounds of the present invention for any length of time contemplated herein.
Psychotherapy
[0243] The compounds, pharmaceutical compositions, and treatment regimes
described herein may
be used in combination with psychotherapy. In one embodiment, methods for the
treatment of
addictive disorder, anxiety disorders, bipolar disorders, and/or depression
may further comprise
psychotherapy.
[0244] Several types of psychotherapy¨or "talk therapy"¨can help people with
depression. In some
embodiments, the regimens are short¨term (e.g., 10 to 20 weeks) and other
regimens are longer¨term
(e.g., 1-10 years), depending on the needs of the individual. Two main types
of psychotherapies¨
cognitive¨behavioral therapy (CBT) and interpersonal therapy (IPT)-have been
shown to be
effective in treating neuropsychiatric disorders (e.g., addictive disorders,
anxiety disorders, bipolar
disorders, and depression).
GABAA Receptors in Disease
[0245] GABAA receptors have a pentameric structure generally comprising two a
subunits and two
[3 subunits with a fifth regulatory subunit. Specific GABAA subunits are
expressed throughout the
brain in distinct spatial and developmental patterns and display different
responses to known
pharmacological modulators. GABAA al variant receptors are believed to be the
major postsynaptic
receptors mediating action of GABA at most inhibitory synapses, and as such
are responsible for not
only the efficacious properties of drugs acting upon GABAA al variant
receptors but also the
sedative effects of these drugs. GABAA a2 and a3 variant receptors are
expressed in the
hippocampus, thalamus, and other CNS locations, and are believed to mediate
the anti-anxiety
effects of the benzodiazepines. at containing GABAA receptors located in the
hippocampus are
thought to play a role in epilepsy. a5 containing GABAA receptors are
expressed in the
hippocampus and are thought to play a role in learning and memory. a4 and a6
containing GABAA
receptors are insensitive to benzodiazepines. Specific GABAA subunits such as
ai and at show
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increased expression in patients with epilepsy, a4 variants of the GABAA
receptor are important in
acting in a negative feedback loop on presynaptic GABA release, where
stimulation of the a4
variants GABAA receptor acts to suppress GABA release.
[0246] The minor "regulatory" subunits c and 0 are expressed in particular CNS
locations such as
the cortex, the substantia nigra, amygdala and hypothalamus. Another minor
regulatory subunit, Tc,
is expressed outside the CNS in the uterus and breast tissue (overexpression
of n has been observed
in breast cancer). Another regulatory subunit, 7 is a component of
benzodiazepine-sensitive GABAA
receptors. The GABAA subunits 72 and 8 are believed to be involved in the
pathologies of certain
monogenetic forms of epilepsy. Also, the GABAA a2 and 8 subunits have been
implicated in
alcohol consumption and addiction. WO 2009/100040.
[0247] The focus of pharmacological intervention in many disorders of the
central and peripheral
nervous system has been on reducing neuronal hyperexcitability. Most agents
currently used to treat
such disorders target synaptic activity in excitatory pathways by, for
example, modulating the
release or activity of excitatory neurotransmitters, potentiating inhibitory
pathways, blocking ion
channels involved in impulse generation, and/or acting as membrane
stabilizers. Conventional
agents and therapeutic approaches for the treatment of central and peripheral
nervous system
disorders thus reduce neuronal excitability and inhibit synaptic firing. One
serious drawback of
these therapies is that they are nonselective and exert their actions on both
normal and abnormal
neuronal populations. This leads to negative and unintended side effects,
which may affect normal
CNS functions, such as cognition, learning and memory, and produce adverse
physiological and
psychological effects in the treated patient. Common side effects include over-
sedation, dizziness,
loss of memory and liver damage. For example, classic anticonvulsant drugs and
anti-nociceptive
drugs decrease excitation or increase inhibition via non-selective GABAergic
drugs (e.g.,
benzodiazepines and barbiturates) indiscriminately act on multiple GABAA
receptor isoforms.
While this yields good efficacy, the non-selective GABAergic drugs cause
undesirable CNS side
effects.
[0248] GABAA receptors are localized at synaptic and extrasynaptic levels.
Whereas synaptic
GABAA receptors are involved in phasic inhibition, extrasynaptic GABAA
receptors are responsible
for tonic inhibition. Tonic inhibition is due to persistent inhibitory
conductance that contributes to
"signal integration" in the brain because it sets the threshold for action
potential generation and
shunts excitatory synaptic inputs. Thus, tonic inhibition plays a crucial role
in regulating neuronal
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excitability because it sets the threshold for action potential generation and
integrates excitatory
signals. This conductance is maintained by "ambient" GABA¨ the amount of
neurotransmitter
present in the extracellular space. Ambient GABA originates from spillover of
the neurotransmitter
released at neighboring synapses, from astrocytes, or from non-vesicular
release. Further, GABAA
receptors are clustered at the synapse and extrasynaptic areas (e.g.,
presynaptic cell). GABAA
receptor clustering acts as an additional regulating factor for tonic
inhibition because clustered
extrasynaptic GABAA receptors can mediate larger tonic currents. Petrini, et
al. (2004) The Journal
of Biological Chemistry 279(44): 45833-45843.
[0249] a4 GABAA receptor variants are primarily located presynaptically or
extrasynaptically (e.g.,
on the pre-synaptic cell). See FIGURE 1. Activation of the sa4 GABAA receptor
variants leads to
hyperpolarization of the pre-synaptic cell, decreasing GABA release and thus
decreasing inhibition
(e.g., agonists specific for oc4 GABAA receptor variants lead to a decrease in
GABA release and
subsequent decrease in the inhibitory signaling). In contrast, inhibition
(antagonism) of ce4 GABAA
receptor variants decreases hyperpolarization of the presynaptic cell, thus
allowing for GABA
release to continue¨ prolonging and strengthening the inhibitory signal to the
postsynaptic cell
(e.g., antagonists specific for or4 GABAA receptor variants lead to an
increase GABA release and
subsequent increase in the inhibitory signaling). In effect antagonists
specific for a4 GABAA
receptor variants achieve physiologic effects similar to those observed in
current GABA agonists.
[0250] The activation of presynaptic GABAA receptors depolarizes the
presynaptic nerve terminals.
The presynaptic actions of neurons can either depress or enhance
neurotransmitter release, processes
called presynaptic inhibition and presynaptic facilitation, respectively. Some
of the best analyzed
instances of presynaptic inhibition and facilitation are in the neurons of
invertebrate animals and in
mechanoreceptor afferent neurons (dorsal root ganglion cells) of vertebrates
studied in dissociated
cell tissue culture. These studies, and those in the intact spinal cord of
mammals, indicate that there
are at least two mechanisms for presynaptic inhibition. One is due to a
synaptically mediated
depression of the Ca2+ channel, leading to a decrease in the influx of Ca2+
into the terminal. The
other is due to an increased conductance to CF that leads to a decrease (or
short-circuiting) in the
height of the action potential in the presynaptic terminal. As a result, less
depolarization is
produced, fewer Ca2+ channels are activated by the action potential, and
therefore, less Ca2+ flows
into the terminals. Activation of GABAA presynaptic receptors is of this
latter type. Antagonism of
the receptor would then lead to presynaptic facilitation. Conversely,
presynaptic facilitation is due
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enhanced influx of Ca2+. The neurotransmitter acts to depress a K.1- channel,
thereby broadening the
action potential and allowing the Ca2+ influx to persist for a longer period
of time.
[0251] Proper neural activity depends on maintaining an appropriate balance
between excitation and
inhibition. Any tipping of the balance too far toward inhibition leads to
sedation, and conversely,
tipping it too far toward excitation may trigger a seizure. For example,
extrasynaptic 8 subunit-
containing GABAA receptors contribute to temporal lobe epilepsy by decreasing
inhibitory input
onto dentate granule cells and increasing the inhibition of inhibitory
interneurons. Peng, et al.
(2004) J. Neurosci. 24: 8629-8639. This increase in the inhibition of the
inhibitory interneurons tips
the balance too far towards excitation by lessening the inhibitory signaling,
leading to seizures.
[0252] Presynaptic actions also tend to occur at points of sensory inflow. For
example, presynaptic
inhibition is found in the retina, spinal cord, and dorsal column nuclei.
Presynaptic actions are
important because they allow selective control of the actions of individual
branches of a neuron.
Axoaxonic synapses can inhibit or facilitate transmitter release by altering
Ca2+ influx. Presynaptic
inhibition may occur as a result of the activity of the postsynaptic cell,
either a presynaptic inhibitory
neuron, or a presynaptic facilitating neuron. In presynaptic inhibition, the
result of the activity of the
presynaptic inhibitor neuron is to cause a depression of the Ca2+ current
accompanying the action
potential of the presynaptic neuron. Because the decreased Ca2+ influx leads
to a reduction in the
amount of neurotransmitter released, the synaptic potential in the
postsynaptic cell is depressed. In
presynaptic facilitation, the activity of the presynaptic facilitating neuron
causes a depression of the
K+ current in the presynaptic neuron leading to an increase in the during of
the action potential and
therefore of the Ca2+ current. Consequently neurotransmitter release is
increased and as a result, so
is the amplitude of the synaptic potential in the postsynaptic cell. Kandel
and Schwartz Principles of
Neural Science 2nd Edition (1985) pages 128-131.
[0253] The cci GABAA variant is expressed at high levels in the hippocampus,
striatum and
thalamus, where it contributes to parasynaptic GABAA receptor mediated tonic
inhibition. Further,
a4 expression is markedly altered by electroshock, alcohol
exposure/withdrawal, steroid withdrawal,
social isolation, and epilepsy. 048 subtypes are modulated by nonbezodiazepine
GABAergic drugs
like steroids, anesthetics, and ethanol. Chandra, et al. (October 10,2006)
Proc. Natl. Acad. Sci.
103(41): 15230-15235.
[0254] Several clinical conditions are thought to arise, in part, from the
imbalance between
neurotransmission of GABA including, but not limited to Huntington's Disease,
Parkinson's disease,
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periodic limb movement disorder (PLMD) (nocturnal myoclonus), spasticity,
epilepsy,
schizophrenia, bipolar disease, and tardive dyskinesia. For instance, GABA
receptors have been
implicated in sleep regulation and rhythmicity as well as the anxiolytic,
amnestic, sedative, and
anesthetic effects of alcohol. Jia, et al. (2008) The Journal of Pharmacology
and Experimental
Therapeutics 326(2): 475-482. Decreased GABA activity appears to contribute to
the pathogenesis
of these diseases. In addition, analgesia and satiety are thought to be
regulated by GABA activity.
Several diseases and conditions are due, at least in part, to an imbalance
between excitation and
inhibition in the central nervous system including but not limited to
addictive disorders, Alzheimer's
Disease, anxiety disorders, attention deficit hyperactivity disorder (ADHD),
autism spectrum
disorders (autism), bipolar disorder, the improvement of cognitive function,
cognitive impairment,
cognitive dysfunction, depression, epilepsy, Huntington's Disease,
inflammatory pain, insomnia,
migraine, migraine with aura, migraine without aura, neuropathic pain,
nociceptive neuralgia,
nociceptive pain, pain, Parkinson's disease, periodic limb movement disorder
(PLMD), personality
disorders, postherpetic neuralgia, psychosis, restless legs syndrome (RLS),
schizophrenia, seizure
disorders, spasticity, tinnitus, and withdrawal syndromes. Therefore,
antagonists specific for or.4
GABAA receptor variants, which lead to an increase GABA release and subsequent
increase in the
inhibitory signaling, may be used to treat these disease and conditions
because they act to restore a
balance between excitation and inhibition in the central nervous system by
increasing inhibition. For
example, antagonists specific for ct4 GABAA receptor variants bind to
extrasynapfic antagonist a.4
GABAA receptor variants preventing their activation by ambient GABA. This
prevents the
hyperpolarization of the presynaptic cell allowing for prolonged GABA release
into the synaptic
cleft that leads to a longer, stronger inhibitory signal. This, in turn,
provides a means by which the
proper balance between excitation and inhibition in the central nervous system
may be restored by
increasing inhibition to counteract an excess of excitation or a lack of
inhibition.
[0255] Compounds described herein selectively antagonize GABAA receptors. In a
preferred
embodiment, the GABAA receptor is a GABAA receptor isoform comprising at least
one cc4. subunit.
In another embodiment, the GABAA receptor is a GABAA receptor isoform
comprising at least one
cc5 subunit. In one embodiment, the invention comprises compositions
comprising compounds
described herein with GABAA receptor antagonist activity. In a further
embodiment, the invention is
drawn to pharmaceutical compositions comprising at least one compound with
GABAA receptor
antagonist activity and a pharmaceutically acceptable excipient. In one
embodiment, compounds
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described herein have GABAA receptor antagonist activity. In another
embodiment, compounds
described herein do not have an effect on GABAB receptors.
[0256] GABAA receptors may be located parasynaptically (e.g., pre- and extra-
synaptically) and
account for control of frequency of IPSCs. Without being committed to a
specific mechanism, the
inventors believe that selected compounds described herein act at parasynaptic
sites by inhibiting the
negative feedback by GABA on the terminal of the synaptic bouton. Parasynaptic
GABAA receptors
act to decrease GABA release when sufficient GABA is present in the synaptic
cleft to bind to and
activate these parasynaptic GABAA receptors (e.g., negative feedback loop). By
inhibiting this
negative feedback loop, compounds described herein may increase the GABA
levels in the synaptic
cleft and decrease neuronal firing. This increase of GABA restores the
appropriate
excitatory/inhibitory balance for normal neuronal activity.
[0257] Many of the compounds described herein may show high selectivity at the
terminals of
GABA interneurons. In vitro electrophysiology studies with selected compounds
described herein
may show selective activity at pre- and/or extra-synaptic (parasynaptic)
terminals indicating
increased GABA release (inhibition). See, e.g., FIGURES 4-8. Without being
bound to a particular
theory of action, the inventors believe that selected compounds described
herein may be GABAA
receptor antagonists that increase the number of inhibitory events as measured
by Inhibitory
Postsynaptic Current (IPSCs). For example, selected compounds described herein
may increase the
frequency of spontaneous IPSCs (a combination of both action potential and
miniature events
releasing GABA) and increase the frequency of miniature IPSCs (miniature
events are due to tonic
release of synaptic vesicles containing GABA into the pre-synaptic space).
[0258] The inventors surprisingly discovered that selected compounds described
herein may
increase GABAA inhibitory drive, since the increased frequency indicates a pre-
synaptic mechanism.
The interval between miniature and spontaneous inhibitory post-synaptic
currents (mIPSCs and
sIPSCs, respectively) events may be substantially decreased in the presence of
selected compounds
described herein. The inventors believe that the pre-synaptic mechanism for
increasing the release
of GABA from the neurons may be due to selected compounds described herein
antagonizing
GABAA receptors on the pre-synaptic cells preventing hyperpolarization of the
pre-terminal cell.
See FIGURE 1. This may then allow for additional GABA to be released into the
synaptic cleft,
leading to longer, and stronger GABA-mediated inhibition.
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[0259] Selected compounds described herein may selectively antagonize specific
GABAA receptor
isoforms (e.g., a.4 variants). a4 GABAA receptor variants are found at
parasynaptic sites and account
for less than 1% of the GABAA receptors in the mammalian brain. The activation
of a.4 GABAA
receptor isoform can inhibit the release of GABA from a GABAergic neuron
(e.g., activation of a cc4
GABAA receptor leads to the hyperpolarization of the synaptic terminal which
slows the release of
GABA from synaptic vesicles and allows GABA clearance mechanisms to lower the
amount of
GABA in the synaptic cleft leading to a decrease of GABA in the synaptic
cleft). Further, the
inventors discovered surprisingly that inhibition of parasynaptic o GABAA
receptor isoforms leads
to an increase in GABA release, which leads to increased inhibitory
stimulation on the post-synaptic
neuron. This specific parasynaptic action supports a possible mechanism for
the lack of CNS
depressant effects (e.g., sedation) demonstrated by compounds described
herein, even at very high
systemic exposure (e.g., after dosages > 100 mg/kg/day). This mechanism of
action is diametrically
opposed to the activation of GABAA receptors by benzodiazepines which work at
low GABA
concentrations. For example, selected compounds described herein may show
selective action on
specific a GABAA receptor isoform (e.g., a4 GABAA variants) generates strong
efficacy in
hyperexcitable states (e.g., epilepsy, migraine, pain) without generating
typical CNS side effects
such as sedation and decreased cognition.
[0260] In particular, compounds of Formulae I-IV described herein may be used
for the regulation,
including prevention, prophylaxis, diagnosis, prognostication, management, and
treatment, of a
range of conditions that involve the GABAA receptor including but not limited
to the disorders
described herein.
[0261] In another embodiment, compounds described herein show selective effect
on GABAA
receptors in the CNS and less side-effects usually associated with agents that
act on GABAA
receptors. For example, compounds described herein exhibit less sedation,
decreased respiration,
decreased cognition, or decreased motor function.
[0262] For example, compounds described herein are effective in humans and
animals to decrease
seizures, decrease pain responses, and decrease migraine. Several of the
compounds described
herein preferentially binds to GABAA receptor subtypes and has an antagonistic
effect on GABAA
receptors that is different from classic benzodiazepine and barbiturate
mechanisms. Unlike some
compounds described herein, several compounds described herein do not act on
the NeK+2C1-
cotransporter (NKCC1 or NKCC2). Some compounds described herein that are
similarly ineffective
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with NKCC1 or NKCC2 are contemplated. Compounds described herein described
herein may not
elicit diuresis. For example, many of compounds described herein may not
increase urine output,
sodium excretion, or potassium excretion.
[0263] Overall, compounds described herein may be well-tolerated
toxicologically and demonstrate
no CNS side effects after oral administration. The inventors surprisingly
discovered that selected
compounds described herein may act to specifically increase neuronal
inhibition via a novel
mechanism of action (not NKCC dependent). The inventors surprisingly
discovered that selected
compounds described herein may act at interneuron terminals, that regulate
neuronal firing, and
therefore, these compounds may inhibit abnormal firing. More specifically,
selected compounds
described herein may increase pre-synaptic inhibition without depressing all
GABA receptors. This
highly selective mechanism of action is novel and contrasts with the broad,
non-specific activity of
benzodiazepines and barbiturates.
[0264] Benzodiazepines and barbiturates are known to be effective but are
poorly tolerated because
these compounds activate most GABAA subtype receptors (e.g., "fire-hose
effect"). In contrast,
selected compounds described herein may enhance inhibition via action at
specific GABAA receptor
subtypes, preferentially o4 variants of GABAA. Due to this selectivity of
selected compounds
described herein may avoid the typical CNS side effects (e.g., sedation)
usually associated with
known GABAergic compounds.
[0265] Further embodiments of the present invention will now be described with
reference to the
following examples. The examples contained herein are offered by way of
illustration and not by
any way of limitation. Exemplary syntheses for compounds according to the
present invention are
provided in, e.g., U.S. Patent Application Publication No. 2007/0149526A1 and
WO 2010/085352.
EXAMPLES
Example 1
3-(Butyl(ethypamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4001
F1715s me--.......---,
0"O
o
General Method A
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0 OH 0 0., 0
H2 N 2 * soci2 H2N.2 N 2 1011 AcCI
N
Me0H CH3CN r THF
8 0 H 8 0 H
N 0 0
= O., 0 0.õ 0 OH
Nyr BH3THF o 2N NaOH
N H2N,9 1101 AD
THF r Me0H
N 0 0 N 0 0 8 0
NTP-4001
Step 1: 3-Butylamino-5-(dimethylaminomethylene-sulfamoyI)-4-phenoxy-benzoic
acid methyl ester.
A round bottom flask was charged with 3-butylamino-4-phenoxy-5-sulfamoyl
benzoic acid (500 mg,
1.37 mmol) and methanol (50 mL). Thionyl chloride (490 mg, 4.12 mmol) was
added slowly at
room temperature and the reaction mixture was heated to 50 C overnight. The
solvent was removed
under reduced pressure and the residue re-dissolved in ethyl acetate and
washed with saturated
sodium bicarbonate solution, water and brine. The organic solvent was removed
under reduced
pressure and the residue purified by flash chromatography to give the product
as pale yellow solid
(512 mg). MS m/z: 379 [M-F 1 r.
Step 2: 3-Butylamino-5-(dimethylaminomethylene-sulfamoyI)-4-phenoxy-benzoic
acid methyl ester.
A reaction flask was charged with 3-butylamino-4-phenoxy-5-sulfamoyl benzoic
acid methyl ester
(509 mg, 1.346 mmol), acetonitrile (4 mL) and N,N-dimethyl formamide dimethyl
acetal (0.19 mL,
1.413 mmol) and stirred at room temperature over night. The solvent was
removed under reduced
pressure and the resultant gummy residue was triturated with ice cold water to
give white solid. The
solid was filtered and dried to give the product (473 mg). MS miz: 434 [M-Fl].
Step 3: 3-(Acetyl-butyl-amino)-5-(dimethylaminomethylene-sulfamoy1)-4-phenoxy-
benzoic acid
methyl ester. A round bottom flask was charged with 3-butylamino-5-
(dimethylaminomethylene-
sulfamoy1)-4-phenoxy-benzoic acid methyl ester (100 mg, 0.230 mmol), acetyl
chloride (0.018 mL,
0.254 mmol), diisopropylethyl amine (0.05 mL), THF (5 mL) and the reaction
stirred at room
temperature for 2 hours. The reaction mixture was poured into water and
extracted with ethyl
acetate. The solvent was removed under reduced pressure and the residue
purified by flash
chromatography to give the product as colorless oil (110 mg). MS m/z: 476 [M-3-
1].
Step 4: 3-(Butyl-ethyl-amino)-5-(dimethylaminomethylene-sulfamoy1)-4-phenoxy-
benzoic acid
methyl ester. A round bottom flask was charged with 3-(acetyl-butyl-amino)-5-
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(dimethylaminomethylene-sulfamoy1)-4-phenoxy-benzoic acid methyl ester (110
mg, 0.23 mmol),
THF (3 mL) and BH3.THF (1.0 M in THF) (4.6 mL, 4.6 mmol) was added slowly. The
reaction was
stirred at room temperature for 1 hour. The reaction was quenched by drop wise
addition of water
and extracted with ethyl acetate. The solvent was removed under reduced
pressure and the residue
purified by flash chromatography to give the product as colorless oil (53 mg).
MS m/z: 462 [M+1].1-.
Step 5: 3-(Butyl-ethyl-amino)-4-phenoxy-5-sulfamoyl-benzoic acid (NTP-4001). A
reaction vial
was charged with 3-(butyl-ethyl-amino)-5-(dimethylaminomethylene-sulfamoy1)-4-
phenoxy-benzoic
acid methyl ester (50 mg, 0.11 mmol), 2N NaOH (3 mL), methanol (3 mL) and the
reaction heated
to 40 C for 3 hours. The solvent was removed under reduced pressure and the
aqueous layer was
acidified with 3N HCI and extracted with ethyl acetate. The solvent was
removed under reduced
pressure to give the desired product (20.3 mg). 1H NMR (300 MHz, DMS0- 4) 6
13.30 (bs, 1H),
8.02 (d, J=1.8 Hz, 1H), 7.72 (d, J=2.18 Hz, 1H), 7.39 (s, 2H), 7.21 (t, J=7.5
Hz, 2H), 6.97 (t, J=7.2
Hz, 1H), 6.76 (d, J=7.8 Hz, 2H), 3.06 (q, J=6.9 & 7.2 Hz, 2H), 2.94 (t, J=6.9
Hz, 2H), 1.17-1.08 (m,
2H), 1.00-0.88 (m, 2H), 0.76-0.68 (m, 6H). MS m/z: 391 [M-1].
Example 2
3-(Butyl(propyl)amino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4002
El2N,
04
The title compound was prepared following General Method A and beginning with
the appropriate
acid chloride in step 3 to give the product as a white solid. MS m/z: 405 [M-
1r.
Example 3
3-(Benzyl(butyl)amino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4003
H''46-06
General Method B
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0 OMe 0 OMe 0 OH
Benzyl bromide
ik K200, k 2N NaOH H2 ;s, N
o,% o CH3CN 01 0 0 0"0 0
reflux Me0H
411 1401
Step 1: Methyl 3-(benzyl(butyl)amino)-5-(N-
((dimethy1amino)methylene)sulfamoy1)-4-
phenoxybenzoate. Methyl 3-(butylamino)-5-(N-
((dimethylamino)methylene)sulfamoy1)-4-
phenoxybenzoate (General Method A, 0.14 g, 0.286 mmol.), benzyl bromide (0.04
mL, 0.343
mmol.), potassium carbonate (60 mg, 0.429 mmol.), and acetonitrile (4 mL) were
charged into a
flask. The mixture was heated to reflux overnight. The excess potassium
carbonate was removed by
filtration over celite. The filtrate was evaporated and purified by
chromatography (n-Hexane/Ethyl
acetate,1/1) to give 0.106 g of the title compound as a white solid.
Step 2: 3-(benzyl(butyl)amino)-4-phenoxy-5-sulfamoylbenzoic acid. Methyl 3-
(benzyl(butyl)amino)-5-(N-((dimethylamino)methylene)sulfamoy1)-4-
phenoxybenzoate (0.106 g,
0.203 mmol.), a 2N aqueous solution of sodium hydroxide (0.3 mL, 0.6 mmol.),
and methanol (2
mL) were charged into a flask. The mixture was heated to 70 C overnight and
then cooled to room
temperature. IN aqueous solution of hydrochloric acid (1 mL) was added to
adjust pH to 2-3, and
the mixture was extracted with ethyl acetate (3 X 2 mL), dried over magnesium
sulfate, and
evaporated to give 0.071 g of title compound as a white solid: 114 NMR (400
MHz, CD30D) 8.23
(d, J= 2.0 Hz, 1H), 7.85 (d, J= 2.0 Hz, 1H), 7.36-7.26 (in, 2H), 7.18-7.06 (m,
4H), 6.92-6.86 (m,
2H), 6.80-6.72 (m, 2H), 2.97 (t, J= 7.4 Hz, 2H), 1.36-1.20 (m, 2H), 1.10-0.96
(m, 2H),0.75 (t, J=
7.4 Hz, 3H). MS m/z: 453 [M-If.
Example 4
3-(Buty1(3-chloratienzyeamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4004
0 0 Lc,
The title compound was prepared following General Method A but beginning with
the appropriate
benzyl bromide in step 2 to give the product as a white solid. 1H NMR (400
MHz, CD30D) (5 8.29
(d, J= 2.0 Hz, 1H), 7.95 (d, J= 2.0 Hz, I H), 7.38-7.26 (in, 2H), 7.18-7.06
(m, 3H), 6.92-6.84 (m,
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2H), 6.80-6.71 (m, 2H), 2.96 (t, J= 7.4 Hz, 2H), 1.38-1.20 (m, 2H), 1.10-0.98
(m, 2H), 0.75 (t, J =
7.4 Hz, 3H). MS nilz: 487 [M-11-.
Example 5
3-(Buty1(4-fluorobenzyparnino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4005
HP.
es..6
The title compound was prepared following General Method A but beginning with
the appropriate
benzyl bromide in step 2 to give the product as a white solid. 11-1 NMR (400
MHz, CD30D) 6 8.26
(d, J = 2.0 Hz, 1H), 7.87 (d, J = 2.0 Hz, 1H), 7.36-7.28 (m, 2H), 7.18-7.16
(m, 1H), 6.92-6.68 (m,
6H), 2.98 (t, J = 7.4 Hz, 2H), 1.33-1.20 (m, 2H), 1.10-0.98 (m, 2H), 0.76 (t,
J = 7.4 Hz, 3H). MS
nilz: 474 [M+1 r.
Example 6
3-(Dimethylamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4006
= OH
H,N,V
000
General Method C
OH
0 0
0 OH 0 OH
0 0
io NaHCO3,
H2N.. g 1101 8(jv=- H N 9 10
NO2 H20 NO
Me0H 2 NO2 AON
NO2
0 01 0 0
0 0
N 0 0
0 0.õ 0 O.
0 OH
Fe, NH4CI N 0 NH2 10 KpC0/,
Mel Ny so 2N NaOH
H2NY 1110
Et0H/H20 ACN
N .õ Me0H
N 0 0 N 0 0
0 0
Step 1: 3-Nitro-4-phenoxy-5-sulfamoyl benzoic acid. A round bottom flask was
charged with 4-
chloro-3-nitro-5-sulfamoyl-benzoic acid (2.0 g, 7.12 mmol), sodium bicarbonate
(2.45 g, 29.2
mmol), phenol (1.47 g, 15.6 mmol) and water (20 mL) and heated at 85 C over
night. The reaction
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mixture was cooled to room temperature and acidified with 3N HCI. The product
precipitated out
which was filtered and dried to give the product as yellow solid (1.9 g). MS
m/z: 337 [M-1].
Step 2: 3-Nitro-4-phenoxy-5-sulfamoy1 benzoic acid methyl ester. A round
bottom flask was
charged with 3-nitro-4-phenoxy-5-sulfamoyl benzoic acid (1.9 g, 5.637 mmol)
and methanol (50
mL). Thionyl chloride (2.012g, 16.91 mmol) was added slowly at room
temperature and the
reaction mixture was heated to 50 C overnight. The solvent was removed under
reduced pressure
and the residue redissolved in ethyl acetate and washed with saturated sodium
bicarbonate solution,
water and brine. The organic solvent was removed under reduced pressure and
the residue purified
by flash chromatography to give the product as pale yellow solid (1.72 g).
Step 3: 3-(Dimethylaminomethylene-sulfamoy1)-5-nitro-4-phenoxy-benzoic acid
methyl ester. A
reaction flask was charged with 3-nitro-4-phenoxy-5-sulfamoyl benzoic acid
methyl ester (1.65 g,
4.68 mmol), acetonitrile (20 mL) and N,N-dimethyl formamide dimethyl acetal
(0.65 mL, 4.917
mmol) and stirred at room temperature over night. The solvent was removed
under reduced pressure
and the resultant gummy residue was treated with ice cold water to give yellow
solid. The solid was
filtered and dried to give the product (1.9 g). MS m/z: 408 [M+1].
Step 4: 5-Amino-3-(dimethylaminomethylene-sulfamoy1)- 4-phenoxy-benzoic acid
methyl ester. A
round bottom flask was charged with 3-(dimethylaminomethylene-sulfamoy1)-5-
nitro- 4-
phenoxy-benzoic acid methyl ester (1.0 g, 2.457 mmol), ethanol (50 mL) and the
reaction mixture
heated to 85 C. Ammonium chloride (1.3 g, 24.57 mmol) in water (25 mL) was
added. Iron powder
(541 mg, 9.828 mmol) was added in three portions 3 minutes apart. The heating
was continued for
another 1 h. The reaction mixture was cooled to 600C and poured into
dichloromethane (150 mL).
The organic layer was separated and washed with water, brine and dried over
sodium sulfate. The
solvents were removed under reduced pressure to give the product as off white
solid (690 mg). MS
m/z: 378[M-1-1]t
Step 5: 3-Dimethylamino-5-(dimethylaminomethylene-sulfamoyI)-4-phenoxy-benzoie
acid methyl
ester. A pressure vial was charged with 5-amino-3-(dimethylaminomethylene-
sulfamoy1)- 4-
phenoxy-benzoic acid methyl ester (200mg, 0.530 mmol), potassium carbonate
(440 mg, 3.18
mmol), methyl iodide (753 mg, 5.305 mmol), acetonitrile (10 mL) and the
reaction heated at 77 C
over night. The reaction was cooled and filtered and washed with ethyl
acetate. The solvent was
removed under reduced pressure and the residue purified by flash
chromatography to give the
product as white solid (70 mg). MS m/z: 406 [M+1]+.
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Step 6: 3-Dimethylamino-4-phenoxy-5-sulfamoyl-benzoic acid (NTP-4006). A
reaction vial was
charged with 3-dimethylamino-5-(dimethylaminomethylene-sulfamoy1)-4-phenoxy-
henzoic acid
methyl ester (70 mg, 0.17 rnmol), 2N NaOH (1 mL), methanol (2 mL) and the
reaction heated to
50 C for 2 hours. The solvent was removed under reduced pressure and the
aqueous layer was
acidified with 3N MCI and extracted with ethyl acetate. The solvent was
removed under reduced
pressure to give the product as white solid (49 mg). 1HNMR (300 MHz, DMS0- d6)
6 13.30 (hs,
1H), 8.00 (d, J=2.1 Hz, 1H), 7.70 (d, J=1.8 Hz, 1H), 7.39 (s, 2H), 7.23 (t,
J=7.8 Hz, 2H), 6.98 (t,
J=7.5 Hz, 1H), 6.76 (d, J.8.4 Hz, 2H), 2.65 (s, 6H). MS m/z: 337 [M+1].
Example 7
3-(Diethylamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4007
0 OH
H2N,9
0
c-]
The title compound was prepared following General Method C but beginning with
the appropriate
alkyl iodide in step 5 to give the product as a white solid (21 mg). MS m/z:
365 [M+1]+.
Example 8
3-(Butyl(cyclopropylmethyl)amino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4008
OH
The title compound was prepared following General Method B but beginning with
the appropriate
alkyl bromide and potassium iodide (1 eq) in step 2 and heating the reaction
of step 2 in a sealed
tube to 100 C for three days. 1H NMR (300 MHz, DMS0- d6) 6 13.28 (bs, 1H),
8.03 (d, J=1.8 Hz,
1H), 7.78 (d, J=2.1 Hz, 1H), 7.38 (s, 2H), 7.21 (t, J=7.5 Hz, 2H), 6.98 (t,
J=7.2 Hz, 1H), 6.76 (d,
J=7.5 Hz, 2H), 3.08 (t, J=6.9 Hz, 2H), 2.90 (d, J=6.6 HZ, 2H), 1.16-1.11 (m,
2H), 0.96-0.89 (m,
2H), 0.70 (t, J=7.5 Hz, 3H),0.59 (m, 1H), 0.30-0.24 (m, 2H), 0.06-0.01 (m,
2H). MS m/z: 419
[114+1]+.
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Example 9
3-(Bis(cyclopropylmethypamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4010
0,15
General Method D
0 O 0 O 0 OH
K2CO3, K õ 0 110r-41
iv.9 rNH 2 ACN, 100 C 2N NaOH H N 95 r'4k
NO 0 NO 0 Me0H 0 0
Step 1: 3-(Bis-cyclopropylmethyl-amino)-5-(dimethylaminomethylene-sulfamoy1)-4-
phenoxy-
benzoic acid methyl ester. A pressure tube was charged with 3-amino-5-
(dimethylaminomethylene-
sulfamoy1)-4-phenoxy benzoic acid methyl ester (250 mg, 0.663 mmol), potassium
carbonate (549
mg, 3.978 mmol), cyclopropyl methyl bromide (269 mg, 1.989 mmol), potassium
iodide (110 mg,
0.663 mmol), acetonitrile (4mL) and heated at 100 C for 3 days. The reaction
gave a mixture of
mono and di-alkylated products by LC/MS. The reaction was cooled and filtered
over celite and the
solvent removed under reduced pressure. The residue was purified by flash
chromatography to give
the product (80 mg). MS m/z: 486.1 [M+1]+.
Step 2: 3-(Bis(cyclopropylmethyl)amino)-4-phenoxy-5-sulfamoylbenzoic acid. A
reaction vial was
charged with 3-(bis-cyclopropylmethyl-amino)-5-(dimethylaminomethylene-
sulfamoy1)-4-phenoxy-
benzoic acid methyl ester (75 mg, 0.15 mmol), 2N NaOH (1 mL), methanol (3 mL)
and the reaction
heated to 50 C for 2 hours. The solvent was removed under reduced pressure and
the aqueous layer
was acidified with 3N HO and extracted with ethyl acetate. The solvent was
removed under
reduced pressure to give the product as white solid (35 mg). 1H NMR (300 MHz,
DMSO- d6) 6
13.28 (bs, 1H), 8.03 (d, J=1.8 Hz, 1H), 7.83 (d, J=2.1 Hz, 1H), 7.37 (s, 2H),
7.21 (t, .1=-7.5 Hz, 2H),
6.97 (t, 3=7.2 Hz, 1H), 6.79 (d, J=7.5 Hz, 2H), 2.95(d, 3=6.3 Hz, 4H), 0.61
(m, 1H), 0.28-0.22 (m,
2H), 0.09-0.04 (m, 2H). MS in/z: 417 [M+1]+.
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Example 10
4-Phenoxy-3-(piperidin-1-yI)-5-sulfamoylbenzoic acid
NTP-4011
0 OH
.2N.140
0 0.6-'
The title compound was prepared following general method C but using 1,5-
diiodopentane in place
of methyl iodide in step 5. MS m/z: 377 [M+1]+.
Example 11
3-(Dibenzylamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4012
0 OH
HANT,? 1401 N *
0 0 ill
The title compound was prepared following general method D but using benzyl
bromide in place of
(bromomethyl)cyclopropoane in step 1. MS m/z: 489 [M+1]+.
Example 12
3-Morpholino-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4013
0 OH
H2N,=9 /--\
S PI 0
8
The title compound was prepared following General Method C but using 1-iodo-2-
(2-iodoethoxy)
ethane in place of methyl iodide in step 5. MS in/z: 379 [M+1]+.
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Example 13
3-(Butyl(pentyl)amino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4014
0 OH
General Method E
0 0., 0 0 0 OH
K2CO3, C5H /11 r
1,1,9 II r KI,9 /¨r-- 2N NaOH
ES
ACN, 150 C. microwave 0 0 411,....\--\\,\ 0 H2"
Me0H 0 0
Step 1: 3-(Butyl-pentyl-amino)-5-(dimethylaminomethylene-sulfamoy1)-4-phenoxy-
benzoic acid
methyl ester. A round bottom flask was charged with 3-butylamino-5-
(dimethylaminomethylene-
sulfamoy1)-4-phenoxy-benzoic acid methyl ester (General Method A step 2, 250
mg, 0.577 mmol),
potassium carbonate (239 mg, 1.732 mmol), iodopentane (228 mg, 1.154 mmol),
acetonitrile (5 mL)
and the reaction heated at 150 C in a microwave reactor for 3 hours. The
reaction was cooled to
room temperature and filtered. The solvent was removed under reduced pressure
and the residue
purified by flash chromatography to give the product as pale yellow solid (140
mg). MS m/z: 504
[M+1]+.
Step 2: 3-(Butyl-pentyl-amino)-4-phenoxy-5-sulfamoyl-benzoic acid (NTP-4014).
A reaction vial
was charged with 3-(butyl-pentyl-amino)-5-(dimethylaminomethylene-sulfamoy1)-4-
phenoxy-
benzoic acid methyl ester (135 mg, 0.27 mmol), 2N NaOH (2 mL), methanol (4 mL)
and the
reaction heated to 50 C for 2 hours. The solvent was removed under reduced
pressure and the
aqueous layer was acidified with 3N HC1 and extracted with ethyl acetate. The
solvent was removed
under reduced pressure to give the product as yellow solid (78 mg). 11-1 NMR
(300 MHz, DMSO-
d6) 6 13.28 (bs, 1H), 7.99 (d, J=1.8 Hz, 1H), 7.71(d, J=2.1 Hz, 1H), 7.34 (s,
2H), 7.20 (t, .1=7.5 Hz,
2H), 6.97 (t, J=7.2 Hz, 1H), 6.72 (d, J=7.5 Hz, 2H), 3.01-2.95 (m, 4H), 1.15-
1.07 (m, 6H), 0.97-
0.89 (m. 4H), 0.76-0.69 (m, 6H). MS in/z: 435 [M+1].
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Example 14
3-Dipentylamino-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4015
0 OH
Fi,N1
General Method F
0 O 0 O
0 OH
Ny 10 NH2 K20.3,05,_õ,1 2N NaOH
H2N.11
N===. ACN, 150 C,rnicrowave N 0 0 Me0H 0 0 46
Step 1: 3-(Dimethylaminomethylene-sulfamoy1)-5-dipentylamino-4-phenoxy-benzoic
acid methyl
ester.
A microwave vial was charged with 3-amino-5-(dimethylaminomethylene-sulfamoy1)-
4-phenoxy
benzoic acid methyl ester (175 mg, 0.501 mmol), potassium carbonate (415 mg,
3.008 mmol),
iodopentane (397 mg, 2.01 mmol) and acetonitrile (4mL) and the reaction heated
at 150 C in a
microwave for 5 hours. A mixture of mono and d-alkylated products was present
by LC/MS. The
reaction was cooled to room temperature and filtered and washed with ethyl
acetate. The solvent
was removed under reduced pressure and the residue purified by flash
chromatography to give the
product as pale yellow solid. The reaction was cooled and filtered over celite
and the solvent
removed under reduced pressure. The residue was purified by flash
chromatography to give the
product (80 mg). MS in/z: 518 [M+11+.
Step 2: 3-(Dipentylamino)-4-phenoxy-5-sulfamoylbenzoic acid (NTP-4015). A
reaction vial was
charged with 3-(dipentylamino)-5-(dimethylaminomethylene-sulfamoy1)-4-phenoxy-
benzoic acid
methyl ester (93 mg, 0.18 mmol), 2N NaOH (1 mL), methanol (2 mL) and the
reaction heated to
50 C for 2 hours. The solvent was removed under reduced pressure and the
aqueous layer was
acidified with 3N HC1 and extracted with ethyl acetate. The solvent was
removed under reduced
pressure to give the product as yellow solid (60 mg). IHNMR (300 MHz, DMS0-
d6) 6 7.99 (d,
J=1.5 Hz, 1H), 7.72 (d, J=1.5 Hz, 1H), 7.37 (s, 2H), 7.20 (t, J.7.5 Hz, 2H),
6.97 (t, J=7.2 Hz, 1H),
6.73 (d, J=7.5 Hz, 2H), 2.99 (t, J=7.5 HZ, 4 H), 1.23-1.07 (m, 8H), 0.95-0.91
(m. 4H), 0.75 (t,
J=7.2 Hz, 6H). MS m/z: 449 [M+11+.
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Example 15
3-(diphenethylamino)-4-phenoxy-5-sulfamoylbenzoic acid
NTP-4016
0"0 01'91 N
The title compound was prepared following General Method F but using (2-
iodoethypbenzene in
place of iodopentane in step 1. MS nilz: 517 [M-F1 r.
Example 16
3-(butyl(phenethyl)amino)-4-phenoxy-5-sulfamaylbenzoic acid
NTP-4017
HO 0
d '0 0
40 40
The title compound was prepared following General Method E but using (2-
iodoethyl)benzene in
place of iodopentane in step 5. MS nilz: 469 [M+1 ].
Following General Method D or with slight modifications thereof, and following
procedures familiar
to one of ordinary skill in the art and the following examples were prepared
using appropriate berizy
bromide in step 1.
Example NTP-Number
Structure M.W.
MS
Number
calculated
nl/Z
[M+ l]
17 NTP-4018
- HO 0 557.48
558
H2N.,s, N CI
o'b 0
110 = a
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18 NTP-4019
HO 0
F 593.43
594
H,N,,s, . N * CI
Orb o
0 140 CI
F
19 NIP-4020
Hy
F 524.54
525
H'19',S,HP'N *
0-'0 0
4 0
F
20 NTP-4021
- HO 0
F 560.52
561
N F
er0 0
* F SF
21 NTP-4022
K0 0
a 593.43
594
,.,2,,,s, 11100 N * F
0'0 0
= 1.1ci F
Example 22
3-(heptan-4-ylamino)-4-phenoxy-5-sulfamoylbenzoic acid
NIP-4023
0 OH 0
01-1
0 OH
0
Zn
9 101 NH4CI
H2N1 0 0 NH, ---- H2N1 THF, Me0H
0 0 NH, Na131-1(0Ac)3
H2N1 0 0 H N
CH3CO2H
lei 101
140
Step 1: 3-amino-4-phenoxy-5-sulfamoylbenzoic acid.
A round bottom flask was charged with 3-nitro-4-phenoxy-5-sulfamoylbenzoic
acid (0.400 g,
0.00118 moles), Ammonium chloride (0.553g. 0.01035 moles), and THF-Me0H (1:1,
10 mL). zinc
(0.676 g, 0.01035 moles) was added under nitrogen and the reaction mixture was
stirred at rt over
night. TLC and LC/MS indicated completion of the reaction. The reaction
mixture was filtered
through Celite and the filter cake was washed with THF-Me0H (1:1). The
filtrate was concentrated
under reduced pressure to give a beige solid (360 mg). MS m/z: 307.0 [M+1].
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Step 2: 3-(heptan-4-ylamino)-4-phenoxy-5-sulfamoylbenzoic acid
To a stirred solution of 3-amino-4-phenoxy-5-sulfamoylbenzoic acid (70.0 mg,
0.00023 moles) in
Acetic acid (2 ml) was added heptan-4-one (0.95 ml, 0.00681 moles) and sodium
sulfate (645 mg,
0.00454 moles), and the mixture was stirred at 70 C for 4h. After cooling to
rt, Sodium
triacetoxyborohydride (0.481 g, 0.00227 moles) was added and the reaction was
stirred at 40 C
overnight. LC-MS indicated that only trace starting material left. The
reaction mixture was diluted
with water and extracted with Et0Ac (2 x). The combined organic layers were
washed with brine,
dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was
purified by silica gel
column (0-80% Et0Ac/hexane) to afford the product as a white foam (30 mg,
38%). MS tn/z: 407.0
[M+1]; Ifl NMR (300 MHz, CDC13 ): 8 8.00 (s, 1H), 7.60 (s, 1H), 7.33 (t, 2H,
.1 = 8.1 Hz), 7.12 (d,
1H, J = 7.5 Hz), 6.96 (d, 2H, J = 8.4 Hz), 4.99 (s, 2H), 3.74 (d, 1H, J = 7.5
Hz), 3.42 (m, 1H), 1.50-
1.00 (m, 8H), 0.83 (t, 6H, J = 7.2 Hz).
Examples 23 and 24
4-Chloro-3-(dimethylamino)-5-sulfamoylbenzoic acid and 4-Chloro-3-
(methylamino)-5-
sulfamoylbenzoic acid
NTP-5001 and NTP-5002
HO 0 HO 0
H2N, . ..., S N Hp. 0 S
N-,
O"b ci 1 cro ci H
General Method G
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0 OH
0 0,, N'--
0 0
SOCl2 .-
-"0-1.0---
Fe, NH4CI
1110
H2N Me0H
N N ii Et0, H20HY 1110
H2N d =ACN
..-- ......., -s
NO2 'S
NO2
NO2
li
0 Cl
0 CI
6 a
0 0.,
0 a.,
0 0,,
K2c03, Mel A.
1 0 SI
1
,
. 0
ACN, reflux 9 0
-I' ...,N..N,g=N..--
NNLg NH2
---N"---N'S N".--
8 a
8 Cl 1
o Cl H
2N NaOH 2N NaOH
Me0H
Me0H
0 OH 0
OH
0 H2N 0
,
H2N õ so ,
'S N
13 H
8 0
a N a 1
Step 1: 4-Chloro-3-nitro-5-sulfamoyl benzoic acid methyl ester. A round bottom
flask was charged
with 4-chloro-3-nitro-5-sulfamoyl benzoic acid (3.0 g, 10.689 mmol) and
methanol (50 mL).
Thionyl chloride (3.82 g, 32.06 mmol) was added slowly at room temperature and
the reaction
mixture was heated to 50 C overnight. The solvent was removed under reduced
pressure and the
residue re-dissolved in ethyl acetate and washed with saturated sodium
bicarbonate solution, water
and brine. The organic solvent was removed under reduced pressure and the
residue purified by
flash chromatography to give the product as pale yellow solid (3.1 g). 1H NMR
(300 MHz, DM50-
d6) (58.72 (d, J=2.1 Hz, 1H), 8.65 (d, J=2.1 Hz, 1H), 8.14 (s, 2H), 3.93 (s,
3H).
Step 2: 4-Chloro-3-(dimethylaminomethylene-sulfamoy1)-5-nitrobenzoic acid
methyl ester. A
reaction flask was charged with 4-chloro-3-nitro-5-sulfamoyl benzoic acid
methyl ester (500 mg,
1.696 mmol), acetonitrile (5 mL) and N,N-dimethyl formamide dimethyl acetal
(0.24 mL, 1.781
mmol) and stirred at room temperature over night. The solvent was removed
under reduced pressure
and the resultant gummy residue was treated with ice cold water to give yellow
solid. The solid was
filtered and dried to give the product (520 mg). MS miz: 350 [M-F 1 ].
Step 3: 3-Amino-4-chloro-5-(dimethylarninomethylene-sulfamoye-benzoic acid
methyl ester. A
round bottom flask was charged with 4-chloro-3-(dimethylaminomethylene-
sulfamoy1)-5-
nitrobenzoic acid methyl ester (500 mg, 1.428 mmol), ethanol (20 mL) and the
reaction mixture
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heated to 85 C. Ammonium chloride (756 mg, 14.28 mmol) in water (10 mL) was
added. Iron
powder (314 mg, 5.71 mmol) was added in three portions 3 minutes apart. The
heating was
continued for another 2 h. The reaction mixture was cooled to 60 C and poured
into
dichloromethane (150 mL). The organic layer was separated and washed with
water, brine and dried
over sodium sulfate. The solvents were removed under reduced pressure to give
the product as off
white solid (340 mg). MS nitz: 320 [M+1]+.
Step 4: 4-Chloro-3-dimethylamino-5-(dimethylaminomethylene-sulfamoy1)-4-
phenoxy-benzoic acid
methyl ester and 4-chloro-3-(methylamino)-5-sulfamoylbenzoic acid. A pressure
tube was charged
with 3-amino-4-chloro-5-(dimethylaminomethylene-sulfamoy1)-benzoic acid methyl
ester (150 mg,
0.469 mmol), potassium carbonate (389 mg, 2.814 mmol), methyl iodide (400 mg,
2.8 mmol),
a.cetonitrile (15 mL) and the reaction heated at 75 C over night. Both the
mono and di-alkylated
products were present by LC/MS. The reaction was cooled and filtered and
washed with ethyl
acetate. The solvent was removed under reduced pressure and the residue
purified by flash
chromatography to give 4-chloro-3-dimethylamino-5-(dimethylaminomethylene-
sulfamoy1)-4-
phenoxy-benzoic acid as pale yellow solid (88 mg) and 4-chloro-3-(methylamino)-
5-
sulfamoylbenzoic acid as yellow solid (35 mg). MS m/z: 348 [M+1] and MS nilz:
334 [M-F1].
Step 5: 4-Chloro-3-dimethylamino-5-sulfamoyl-benzoic acid (NTP-5001). A
reaction vial was
charged with 4-chloro-3-dimethylamino-5-(dimethylaminomethylene-sulfamoy1)-4-
phenoxy-benzoic
acid methyl ester (85 mg, 0.247 mmol), 2N NaOH (2 mL), methanol (4 mL) and the
reaction heated
to 50 C for 2 hours. The solvent was removed under reduced pressure and the
aqueous layer was
acidified with 3N HC1 and extracted with ethyl acetate. The solvent was
removed under reduced
pressure to give the product as white solid (49 mg). ill NMR (300 MHz, DMS0-
d6) 6 13.51 (bs,
1H), 8.16 (d, J.2.1 Hz, 1H), 7.81 (d, J.1.8 Hz, 1H), 7.70 (s, 2H), 2.79 (s,
6H). MS in/z: 279 [M+1r.
Step 6: 4-Chloro-3-methylamino-5-sulfamoyl-benzoic acid (NTP-5002). The
product was obtained
following the procedure described for step 5 and starting with 4-chloro-3-
(methylamino)-5-
sulfamoylbenzoic acid to give the title compound as white solid (16 mg). Ili
NMR (300 MHz,
DMS0- d6) 6 13.51 (bs, 1H), 8.45 (d, J.2.1 Hz, 1H), 7.76 (d, J.-1.8 Hz, 1H),
7.34 (s, 2H), 5.69 (m,
1H), 2.80 (d, 7.5.1 Hz, 3H). MS in/z: 263 [M-1 ].
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Examples 25 and 26
4-Chloro-3-(diethylamino)-5-sulfamoylbenzoic acid and 4-Chloro-3-(ethylamino)-
5-
sulfamoylbenzoic acid
NTP-5003 and NTP-5004
HO 0 HO 0
H,N,* H2N ,s Nr...õ.
0-0 ) olµb H
The title compounds were prepared following General Method G and using the
appropriate alkyl
iodide in step 4. 4-chloro-3-(diethylamino)-5-sulfamoylbenzoic acid: MS nilz:
307 [M+1]. 4-
chloro-3-(ethylamino)-5-sulfamoylbenzoic acid: MS miz: 279 [M+1]*.
Example 27
3-(Butylamino)-4-chloro-5-sulfamoylbenzoic acid
NTP-5005
Ho o
H2N
e,",µ
00 ci H
General Method H
O... o o O
= o
o
BH3.THF
N N NH2 DIEA, THF =0
THF 2
0 CI 0 CI
0 CI
0 OH
2N NaOH
0 ao
H2N II
Me0H
0 CI
Step 1: 3-Butyrylamino-4-chloro-5-(dimethylaminomethylene-sulfamoy1)-benzoic
acid methyl ester.
A round bottom flask was charged with 3-amino-4-chloro-5-
(dimethylaminomethylene-sulfamoy1)-
benzoic acid methyl ester (Method G, step 3, 500 mg, 1.56 mmol), butyryl
chloride (0.20 mL, 1.876
mmol), diisopropylethyl amine (0.1 mL), THF (5 mL) and the reaction stirred at
room temperature
for 2 hours. The reaction mixture was poured into water and extracted with
ethyl acetate. The
solvent was removed under reduced pressure and the residue purified by flash
chromatography to
give the product as light brown oil (650 mg). MS nilz: 390.2 [M+1]+.
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Step 2: 3-Butylamino-4-chloro-5-(dimethylaminomethylene-sulfamoy1)-benzoic
acid methyl ester: A
round bottom flask was charged with 3-butyrylamino-4-chloro-5-
(dimethylaminomethylene-
sulfamoy1)-benzoic acid methyl ester (600 mg, 1.539 mmol), THF (10 mL) and
BH3.THF (1.0 M in
THF) (7.69 mL, 7.69 mmol) was added slowly. The reaction was stirred at room
temperature for 1
hour. The reaction was quenched by drop wise addition of water and extracted
with ethyl acetate.
The solvent was removed under reduced pressure and the residue purified by
flash chromatography
to give the product as pale yellow solid (340 mg). MS mlz: 376.3 [M+1] .
Step 3: 3-Butylamino-4-chloro-5-sulfamoyl-benzoic acid (NTP-5005).
A reaction vial was charged with 3-butylamino-4-chloro-5-
(dimethylaminomethylene-sulfamoy1)-
benzoic acid methyl ester (340 mg, 0.90 mmol), 2N NaOH (5 mL), methanol (5 mL)
and the
reaction heated to 50 C for 2 hours. The solvent was removed under reduced
pressure and the
aqueous layer was acidified with 3N HCI and extracted with ethyl acetate. The
solvent was removed
under reduced pressure to give the product as white solid (105 mg). 1HNMR (300
MHz, DMSO-
d6) 6 13.34 (bs, 1H), 7.72 (d, J=2.1 Hz, 1H), 7.59 (s, 2H), 7.33 (d, J=1.8 Hz,
1H) 5.99 (t, J=5.4 Hz,
1H), 3.20 (t, J=6.3 Hz, 2H), 1.59-1.52 (m, 2H), 1.40-1.1.33 (m, 2H), 0.92 (t,
J=6.6 Hz, 3H). MS
m/z: 307.1[M-1-1].
Example 28
4-Chloro-3-(dibutylamino)-5-sulfamoylbenzoic acid
NTP-5006
HO 0
H211;s,
do cr
The title compound was prepared following General Method G and using the
appropriate alkyl
iodide in step 4. 11-1 NMR (300 MHz, CD30D) 6 8.34 (m, 1H), 7.96 (m, 1H), 3.14
(t, J=5.7 Hz, 4H),
1.48-1.45(m, 4H), 1.38-1.31 (m, 4H), 0.90 (t, J=6.9 Hz, 6H). MS in/z: 363 [M-i-
1].
Example 29
2-(Dibutylamino)-6-sulfamoylbipheny1-4-carboxylic acid
NTP-5007
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0 OH
H2N1 rf"--/----
8
General Method I
HO,B,OH
0 O.,
= (:)
0 O.,
N N 'S 0 NO2
POPd, dioxan, 900C ,
N 9 10 NO2Et0H,
H2OH203' Ill N1 Fe, NH4GI
NH2
0 GI
8
O.
.
K2CO3, Bel
I 0 ioH2N
2N NaOH
AGN, 100 G, 24 h
8
,9 0
N
Step 1: 2-(Dimethylaminomethylene-sulfamoy1)-6-nitro-bipheny1-4-carboxylic
acid methyl ester. A
reaction vial was charged with 4-chloro-3-(dimethylaminomethylene-sulfamoy1)-5-
nitrobenzoic acid
methyl ester (Method G, step 2, 220 mg, 0.630 mmol), phenyl boronic acid (115
mg, 0.945 mmol),
potassium carbonate (261 mg, 1.89 mmol), [(t-Bu)2P(OH)]2 PdC12 (P0Pd) (3 mg,
0.0063 mmol) and
1,4-dioxane (3 mL). The vial was evacuated with vacuum and flushed with
nitrogen. The reaction
was then heated at 90 C overnight. The reaction mixture was cooled to room
temperature, diluted
with ethyl acetate and washed with water. The solvent was removed under
reduced pressure and the
residue purified by flash chromatography to give the product in 60% purity
which was taken to next
step without further purification (90 mg). MS m/z: 392 [M+1
Step 2: 6-Amino-2-(dimethylaminomethylene-sulfamoy1)- biphenyl-4-carboxylic
acid methyl ester.
A round bottom flask was charged with 2-(dimethylaminomethylene-sulfamoy1)-6-
nitro-bipheny1-4-
carboxylic acid methyl ester (90 mg, 0.229 mmol), ethanol (10 mL) and the
reaction mixture heated
to 85 C. Ammonium chloride (122 mg, 2.29 mmol) in water (3 mL) was added. Iron
powder (51
mg, 0.917 mmol) was added in three portions 3 minutes apart. The heating was
continued for
another 2 h. The reaction mixture was cooled to 60 C and poured into
dichloromethane (100 mL).
The organic layer was separated and washed with water, brine and dried over
sodium sulfate. The
solvents were removed under reduced pressure and the residue purified by flash
chromatography to
give the product as off white solid (60 mg). MS m/z: 362 [M+1]+.
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Step 3: 6-Dibutylamino-2-(dimethylaminomethylene-sulfamoy1)-bipheny1-4-
carboxylic acid methyl
ester. A pressure tube was charged with 6-amino-2-(dimethylaminomethylene-
sulfamoy1)-
bipheny1-4-carboxylic acid methyl ester (60 mg, 0.166 mmol), potassium
carbonate (138 mg, 0.996
mmol), butyl iodide (122 mg, 0.664 mmol), acetonitrile (5 mL) and the reaction
heated at 120 C
over night. The reaction gave a mixture of mono and di-alkylated products. The
reaction was
cooled and filtered and washed with ethyl acetate. The solvent was removed
under reduced pressure
and the residue purified by flash chromatography to give the product (28 mg).
MS m/z: 474 [M+1]+.
Step 4: 6-Dibutylamino-2-sulfamoyl-biphenyl-4-carboxylic acid (NTP-5007). A
reaction vial was
charged with 6-dibutylamino-2-(dimethylaminomethylene-sulfamoyI)-biphenyl-4-
carboxylic acid
methyl ester (28 mg, 0. 059 mmol), 2N NaOH (1 mL), methanol (3 mL) and the
reaction heated to
50 C for 2 hours. The solvent was removed under reduced pressure and the
aqueous layer was
acidified with 3N HCI and extracted with ethyl acetate. The solvent was
removed under reduced
pressure to give the product as yellow solid (16 mg). 11-1 NMR (300 MHz, DMS0-
d6) 6 13.41 (bs,
1H), 8.25 (d, J=1.5 Hz, 1H), 7.83 (d, J=1.5 Hz, 1H), 7.38-7.28 (m, 3H), 7.19
(d, J=6.9, 2H), 7.00 (s,
2H), 2.68 (m, 4H), 1.05-1.03 (m, 8H), 0.74 (t, J=6.6 Hz, 6H). El MS m/z: 405
[M-1-1]*.
Example 30
3-(Dibutylamino)-4-(3-fluorophenoxy)-5-sulfamoylbenzoie acid
NTP-5009
= OH
N
General Method J
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0 OH HO 11"
0 OH
0 0..õ,
0 F
0
H2N,O
H2N,
9
NaHCO3
9
CI
N
NO2 H2O
NO2 Me1/4_,H H2N
0 0....-
NO2
ACN
NO2
8 0
o o
o o
N 0 0
F
F
0 0.,
0 O
0 OH
10% Pd/C
H2N K2CO2, Bul
NY
2N NaOH
9
iõ--/¨
Et0H
'n
NH,
ACN, 100 C, 4 daysi.-
s
N 0 0
,N.., 0
0
0
Step 1: 4-(4-Fluorophenoxy)-3-nitro-5-sulfamoylbenzoic acid. To a stirred
solution of 4-Chloro-3-
nitro-5-sulfamoylbenzoic acid (1 g, 3.56 mmol) in water was added 4-
fluorophenol (0.8 g, 7.12
mmol) and NaHCO3 (1.2 g, 14.24 mmol) at 0 C. The mixture was stirred at 100 C
for 12 hours,
then it was cooled to room temperature. The mixture was made acidic with 6 N
HC1. The light
yellow solid came out, then it was filtered to give a crude 4-(4-
fluorophenoxy)-3-nitro-5-sulfamoyl-
benzoic acid, which was used in the next reaction without further
purification.
Step 2: Methyl 4-(4-fluorophenoxy)-3-nitro-5-sulfamoylbenzoate. A solution of
4-(4-
fluorophenoxy)-3-nitro-5-sulfamoylbenzoic acid (0.97 g, 2.72 mmol) in methanol
was added acetyl
chloride (0.42 g, 5.38 mmol) dropwise at 0 C. The mixture was stirred at 60 C
for 12 hours. When
all starting material disappeared, water was added slowly at 0 C. Methanol was
removed, then the
aqueous solution was extracted with ethyl acetate. The organic layers were
combined, dried over
anhydrous MgSO4, and evaporated. The residue was purified by flash
chromatography (n-
hexane/ethyl acetate = 1/1) to afford methyl 4-(4-fluorophenoxy)-3-nitro-5-
sulfamoylbenzoate (0.37
g).
Step 3: Methyl 3-(N-((dimethylamino)methylene)sulfamoy1)-4-(4-fluorophenoxy)-5-
nitrobenzoate.
To a solution of methyl 4-(4-fluorophenoxy)-3-nitro-5-sulfamoylbenzoate (0.37
g, 1 mmol) in
acetonitrile was added N,N-Dimethylformamide dimethyl acetal (DMF/DMA, 0.178g,
1.5 mmol) at
room temperature for 30 minutes. The reaction mixture was stirred at room
temperature for 1 hour,
and then quenched with water. The resulting mixture was extracted with ethyl
acetate. The organic
layer was separated and washed with 0.1 M HCI, dried over anhydrous MgSO4, and
evaporated. The
residue was purified by flash chromatography (n-hexane/ethyl acetate = 5/1) to
afford the title
compound (0.27 g).
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Step 4: Methyl 3-amino-5-(N-((dimethylamino)methylene)sulfamoy1)-4-(4-
fluorophenoxy)benzoate.
A mixture of 3-(N-((dimethylamino)methylene)sulfamoy1)-4-(4-fluorophenoxy)-5-
nitrobenzoate
(190 mg, 0.447 mmol) and 10% Pd/C (19 mg, 10% w/w) in ethyl alcohol was
stirred at 60 C under
H2 atmosphere. After 3 hours the mixture was passed through a celite to remove
the catalyst. The
filtered catalyst was washed with ethyl alcohol (2x10 mL). The combined
filtrates were washed
with H20 (2x30 mL) and brine (30 mL), dried over anhydrous MgSO4, and
evaporated under
reduced pressure. The residue was purified by flash chromatography (n-
hexane/ethyl acetate = 1/1)
to afford the title compound (30 mg).
11-1 NMR (400 MHz, DMSO-d6) 6 7.91 (s, 1H), 7.71 (s, 1H), 7.63 (s, 1H), 7.18-
7.11 (m, 2H), 6.74-
6.71 (m, 2H), 5.37 (s, 2H), 3.86 (s, 3H), 2.92 (s, 3H), 2.55 (s, 3H).
Step 5: 3-Dibutylamino-5-(dimethylaminomethylene-sulfamoy1)-4-(4-fluoro-
phenoxy)-benzoic acid
methyl ester. A pressure tube was charged with 3-amino-
5(dimethylaminomethylene-sulfamoy1)-4-
(4-fluorophenoxy)-benzoic acid methyl ester (140 mg, 0.352 mmol), potassium
carbonate (292 mg,
2.115 mmol), butyl iodide (259 mg, 1.408 mmol), acetonitrile (5 mL) and the
reaction heated at
100 C for 4 days. The reaction gave a mixture of mono and di-alkylated
products. The reaction was
cooled and filtered and the solid washed with ethyl acetate. The filtrate was
concentrated under
reduced pressure and the residue purified by flash chromatography to give the
product (51.5 mg).
MS mtz: 508 [M-f-1 r.
Step 6: 3-Dibutylamino-4-(4-fluoro-phenoxy)-5 -sulfamoyl-benzoic acid (NTP-
5009). A reaction
vial was charged with 3-dibutylamino-5-(dimethylaminomethylene-sulfamoy1)-4-(4-
fluoro-
phenoxy)-benzoic acid methyl ester (51 mg, 0.101 mmol), 2N NaOH (1 mL),
methanol (3 mL) and
the reaction heated to 50 C for 2 hours. The solvent was removed under reduced
pressure and the
aqueous layer was acidified with 3N HCI and extracted with ethyl acetate. The
solvent was removed
under reduced pressure to give the product as pale yellow solid (37 mg). 1H
NMR (300 MHz,
DMS0- d6) (5 13.30 (bs, 1H), 8.00 (d, J=2.I Hz, IH), 7.73 (d, J=2.1 Hz, 1H),
7.42 (s, 2H), 7.08-7.03
(m, 2H), 6.78-6.73 (m, 2H), 3.00 (t, 3=7.2 Hz, 4H), 1.21-1.11 (m, 4H), 1.01-
0.91 (m, 4H), 0.73 (t,
J=7.2Hz, 6H). MS m/z: 439 [M+1]t
Following General Method .1' or with slight modifications thereof, and
following procedures familiar
to one of ordinary skill in the art and the following examples were prepared
from commercially
available reagents.
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Example NTP-
Number
Structure
MW. MS
Number
calculated nilz
[M+1
31 NTP-
5008
0 OH
438.51 439
HAI 40 0 0
32 NTP-
5010
OR
450.55 451
H2NI
33 NTP-
5011 H,N3
O OCH ¨o
434.55 435
34 NTP-
5012
0 OH
450.55 451
a2N,V 1110g0
35 NTP-
5013
0 OH
454.97 455
H2Ny 40 0 0 N
36 NTP-
5014
= OH
450.55 451
H2N1 8o
37 NTP-
5015
0 OH o-
445.53 446
8
38 NTP-
5016
0 OH CN
489.41 489
H2N4 8 0 N.F"---7--
CI CI
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39 NTP-5017
= - 01-1
454.97 455
"2¶,& 0 a
b
01
40 NTP-5018
0 OH
398.86 399
H2N1 9 = /¨/¨ N
O 0 H
CI
41 NTP-5019
0 OH
378.44 379
H2N1 O 1. 0 " N
401
42 NTP-5020
0 OH
382.41 383
H2N1 c? N /-7¨
O 0 H
140
Example 43
3-(Dibutylamino)-5-(N,N-dimethylsulfamoy1)-4-phenoxybenzoic acid
NTP-6001
CO,F1
0-06 LI,
General Method K
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0 OH
0 OH
0 O.,
\ /
N
H
S0012
NaHco, Phenol
,p, 0
1
.. It1,9 10
a
Me0H ki? ill
NO2 -- ..., s
s NO2
NO2 DMSO,
0 Cl
8 a
8 Cl
0 0.,
0 00 0
. 0
.
Fe/NH401 1 0 0
1 9 ip
_i.._
_19 is x...õ--___ BH3.THF..._
NO2 Et0H/H20 .--NLS
NH 2 THF
S N
THF
H
0 0,
8 0,
8
Ir
0 0.,
0 OH
NaH, BuBr I 0 101
...-. r14,9 SO ,--7---
N= ,N.,11
N THF,
S N
H
0 0 0
8 0 i\---\-
-
Ir
Step 1: 4-Chloro-3-dimethylsulfamoy1-5-nitro-benzoic acid. A round bottom
flask was charged with
4-chloro-3-chlorosulfony1-5-nitro-benzoic acid (500g, 1.66 mmol),
dimethylamine (2.0 M in THF, I
mL, 1.99 mmol), diisopropylethylamine (0.36 mL, 1.99 mmol) and THF (3 mL) and
the reaction
was stirred at 45 C overnight. The solvent was removed under reduced pressure
and the residue
dissolved in ethyl acetate and washed with water, brine and dried over Na2SO4.
The solvent was
removed under reduced pressure to give the product as yellow solid (360 mg)
which was used in the
next reaction without further purification. MS in/z: 307 EM-1f.
Step 2: 4-Chloro-3-dimethylsulfamoy1-5-nitro-benzoic acid methyl ester. A
round bottom flask was
charged with 4-chloro-3-dimethylsulfamoy1-5-nitro-benzoic acid (360 mg, 1.16
mmol) and methanol
(10 mL). Thionyl chloride (152 mg, 1.283 mmol) was added slowly at room
temperature and the
reaction mixture was heated to 50 C overnight. The solvent was removed under
reduced pressure
and the residue re-dissolved in ethyl acetate and washed with saturated sodium
bicarbonate solution,
water and brine. The organic solvent was removed under reduced pressure and
the residue purified
by flash chromatography to give the product as yellow solid which was used
directly in the next
reaction (340 mg).
Step 3: 3-Dimethylsulfamoy1-5-nitro-4-phenoxy-benzoic acid methyl ester. A
round bottom flask
was charged with 4-chloro-3-dimethylsulfamoy1-5-nitro-benzoic acid methyl
ester (340 mg, 1.054
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mmol), sodium bicarbonate (354 mg, 4.217 mmol), phenol (198 mg, 2.108 mmol)
and DMSO (10
mL) and the reaction mixture heated to 80 C overnight. The reaction was cooled
to room
temperature and quenched with water. The product which precipitated out was
filtered and
dissolved in ethyl acetate and washed with brine. The solvent was removed
under reduced pressure
to give the product as pale yellow solid (340 mg). 11-1 NMR (300 MHz, DMS0-
d6) c5 8.71 (d, J=1.8
Hz, 1H), 8.63 (d, J=2.1 Hz, 1H), 7.32 (t, J=5.7 Hz, 2H), 7.10 (t, J=7.5 Hz,
1H), 6.92 (d, J=8.4 Hz,
2H), 3.95 (s, 3H), 2.79 (s, 6H).
Step 4: 3-Amino-5-dimethylsulfamoy1-4-phenoxy-benzoic acid methyl ester. A
round bottom flask
was charged with 3-dimethylsulfamoy1-5-nitro-4-phenoxy-benzoic acid methyl
ester (340 mg, 0.894
mmol), ethanol (10 mL) and the reaction mixture heated to 85 C. Ammonium
chloride (475 mg,
8.94 mmol) in water (5 mL) was added. Iron powder (197 mg, 3.578 mmol) was
added in three
portions 3 minutes apart. The heating was continued for another 1 h. The
reaction mixture was
cooled to 60 C and poured into dichloromethane (150 mL). The organic layer was
separated and
washed with water, brine and dried over sodium sulfate. The solvents were
removed under reduced
pressure to give the product as pale yellow solid (300 mg). MS m/z: 351
[M+1]+.
Step 5: 3-Butyrylamino-5-dimethylsulfamoy1-4-phenoxy-benzoic acid methyl
ester. A round bottom
flask was charged with 3-amino-5-dimethylsulfamoy1-4-phenoxy-benzoic acid
methyl ester (330 mg,
0.942 mmol), butyryl chloride (0.12 mL, 1.13 mmol), diisopropylethyl amine
(0.1 mL), THF (5 mL)
and the reaction stirred at room temperature for 2 hours. The reaction mixture
was poured into water
and extracted with ethyl acetate. The solvent was removed under reduced
pressure and the residue
purified by flash chromatography to give the product as light brown oil (400
mg). MS m/z: 421
[M+1]+.
Step 6: 3-Butylarnino- 5-dimethylsulfamoy1-4-phenoxy-benzoic acid methyl
ester. A round bottom
flask was charged with 3-butyrylamino-5-dimethylsulfamoy1-4-phenoxy-benzoic
acid methyl ester
(330 mg, 0.785 mmol), THF (3 mL) and BH3.THF (1.0 M in THF) (4 mL, 3.928 mmol)
was added
slowly. The reaction was stirred at room temperature for 1 hour. The reaction
was quenched by
drop wise addition of water and extracted with ethyl acetate. The solvent was
removed under
reduced pressure and the residue purified by flash chromatography to give the
product as pale yellow
solid (160 mg). MS m/z: 407 [M+ 1 ]t
Step 7: 3-Dibutylamino-5-dimethylsulfamoy1-4-phenoxy-benzoic acid (NTP-6001).
A reaction vial
was charged with 3-butylamino-5-dimethylsulfamoy1-4-phenoxy-benzoic acid
methyl ester (30 mg,
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0.073 mmol), THF (2 mL), sodium hydride (95%) (3.5 mg, 0.147 rnmol) and butyl
bromide (16 mg,
0.147 rnmol) and the reaction stirred at 500 over night. The reaction was
quenched with water and
acidified with 3N HCI and extracted with ethyl acetate. The solvent was
removed under reduced
pressure to give the product as colorless viscous oil (17 mg). 11-INMR (300
MHz, DMS0- dis) 6
7.94 (d, J=1.8 Hz, 1H), 7.78 (d, J=1.8 Hz, 1H), 7.23 (t, J=7.5 Hz, 2H), 6.99
(t, J=7.2 Hz, 1H), 6.67
(d, J=7.5 Hz, 2H), 3.02 (t, J=7.2 Hz, 4H), 2.70 (s, 6H), 1.23-1.1.11(m, 4H),
0.98-0.93 (m, 4H), 0.72
(t, J=7.5 Hz, 6H). MS ink: 449 [M+1].
Example 44
3-(Dibutylamino)-5-(morpholinosulfony1)-4-phenoxybenzoic acid
NTP-6002
co,n
(04N---------
cro o LL
6
Following General Method K or with slight modifications thereof, and following
procedures familiar
to one of ordinary skill in the art and the title compound was prepared from
commercially available
reagents. MS m/z: 491 [M+1]+.
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Example 45
3-(Butylamino)-5-(N,N-dimethylsulfamoyI)-4-phenoxybenzoic acid
NTP-6003
CO2H
0"0 o H
A reaction vial was charged with 3-butylamino-5-dimethylsulfamoy1-4-phenoxy-
benzoic acid
methyl ester (General Method K, step 6, 130 mg, 0319 mmol), methanol (2 mL),
THF (I mL) and
1N LiOH (1 mL) and the reaction stirred room temperature 2 hours. The solvent
was removed under
reduced pressure and the aqueous layer was acidified with 3N HCI. The mixture
was extracted with
ethyl acetate and the solvent removed under reduced pressure to give the
product as white solid (93
mg). Ili NMR (300 MHz, DMS0- d6) 613.25 (bs, 1H), 7.60 (d, J=2.1 Hz, 1H), 7.46
(d, J=2.1 Hz,
1H), 7.27 (t, J=7.5 Hz, 2H), 7.01 (t, J=7.2 Hz, 1H), 6.75 (d, J=7.5 Hz, 2H),
5.18 (t, J= 5.4 Hz, 1H),
3.09-3.03(m, 2H), 1.39-1.34 (m, 2H), 0.77 (t, J=7.2 Hz, 6H). MS m/z: 393 [M-
Fl].
Example 46
3-(Butylamino)-5-(morpholinosulfony1)-4-phenoxybenzoic acid
NTP-6004
ON, 0 H
The title compound was prepared in a manner similar to that used to prepare
example 33.
MS in/z: 435.1 [M+1]+.
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Example 47
3-(N-acetylsulfamoy1)-5-(dibutylarnino)-4-phenoxybenzoic acid
NTP-6005
N.
0 0 0
Methyl 3-(dibutylamino)-4-phenoxy-5-sulfamoylbenzoate (0.16 g, 0.37 mmole),
and Et3N (0.1 mL,
0.74 mmole), and Ac20 (0.04 mL, 0.44 mmole), and CH2C12 (2 mL) were charged
into a flask. The
mixture was stirred at room temperature for 2 hours. The reaction solution was
quenched with water
(10 mL). The methanol was removed on rotavapor, and the aqueous solution was
extracted with
Et0Ac (2 X 10 mL). The organic layers were combined, dried over MgSO4, and
evaporated. The
residue was purified by flash column (n-hexane/ Et0Ac :1/1) to yield 0.11 g of
a white solid. The
solid (0.105 g, 0.221 mmol), 2N NaOH (0.22 mL, 0.442 mmol), and Me0H (2 mL)
were charged
into a flask. The mixture was heated to 40 C for 2 hours. The reaction
solution was neutralized
with 1N HC1 (1 mL), and the mixture was extracted with Et0Ac, dried over
MgSO4, and evaporated
to yield 0.04 g of the title compound as a white solid. Ili NMR (400 MHz,
CD30D) (5 8.28 (d, J =
2.0 Hz, 1H), 7.94 (d, J= 2.0 Hz, 1H), 7.35-7.20 (m, 2H), 7.10-7.02 (m, 1H),
6.80-6.70 (m, 1H),
3.20-3.02 (m, 4H), 1.54 (s, 3H), 1.40-1.15 (m, 4H), 1.10-1.09 (m, 4H), 0.79
(t, J= 7.2 Hz, 6H).
MS m/z: 463 [M+ 1I
Examples 48 and 49
3-(Dibutylamino)-5-(N-ethylsulfamoy1)-4-phenoxybenzoic acid and 3-
(Dibutylamino)-5-(N,N-
diethylsulfamoy1)-4-phenoxybenzoic acid
NTP-6006 and NTP-6008
= OH
FNIY 401 9
0 0 oO
General Method L
0 OH 0 01-1
0 OH
H2N,Ii0 1110 THF 0
0 0 0 0
0
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A round bottom flask was charged with 3-dibutylamino-4-phenoxy-5-sulfamoyl-
benzoic acid (1.0g,
2.38 mmol), sodium hydride (95%) (171 mg, 7.134 mmol) and THF (15 mL). The
reaction mixture
was stirred for 15 minutes at room temperature and ethyl iodide (1.12 g, 7.134
mmol) was added and
the reaction mixture was heated to 50 C overnight. The reaction mixture showed
diethyl, mono
ethyl and ethyl ester products by LCMS. The reaction mixture was quenched by
addition of water
and extracted with ethyl acetate. The organic layer was then washed with 3N
HCI, water, brine and
dried over Na2SO4. The solvent was removed under reduced pressure and the
residue purified by
flash chromatography to give 3-(dibutylamino)-5-(N-ethylsulfamoy1)-4-
phenoxybenzoic acid (28
mg) and 3-(dibutylamino)-5-(N,N-diethylsulfamoy1)-4-phenoxybenzoic acid (132
mg) as white
solids.
3-(Dibutylamino)-5-(N-ethylsulfamoy1)-4-phenoxybenzoic acid: MS m/z: 449 [M+
1].
3-(dibutylamino)-5-(N,N-diethylsulfamoy1)-4-phenoxybenzoic acid: MS m/z: 477.2
[M+11+.
Following General Method L or with slight modifications thereof, and following
procedures familiar
to one of ordinary skill in the art the following examples were prepared from
commercially available
reagents.
Example NTP-Number Structure M.W.
Mass
Number
calculated Spec.
m/z
[M-1-1]
50 NTP-6007 00:01: 420.52
421
000
51 NTP-6009 . OH 474.61
475
Co 40
0OO
52 NTP-6010 0 OH 528.70
529
k--11q,9 40
0 0
VP
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53 NTP-
6011
0 OH
474.61 475
tO
54 NTP-
6012
0 OH
434.55 435
Li
OO0
55 NTP-
6013
H015
488.64 489
00
56 NTP-
6014
FlOy0
510.64 511
o
57 NTP-
6015
T02H
621.48 622
o'b 1,
1110 c,
58 NTP-
6016
0 10 H
460.59
01- Fcl'IrN 0
Example 59
3-(Dibutylamino)-5-(morpholine-4-carbony1)-2-phenoxybenzenesulfonamide
NTP-3032
0
4
0
General Method M
0 OH
0
H2N.
HN 0 EDCI, HOBt
H2 9 1110
8 a
Ei3N, DCM
o o
o oc R.T.
40
40
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To a solution of 3-(dibutylamino)-4-phenoxy-5-sulfamoylbenzoic acid (100 mg,
0.238 mmol) in
dichloromethane was added morpholine (41 mg, 0.476 mmol), triethylamine (48
mg, 0.476 mmol),
1-(3-dimethylaminopropy1)-3-ethylcarbodiimide hydrochloride (54.7 mg, 0.286
mmol), and 1-
hydroxybenzotriazole (48 mg, 0.357 mmol) at 0 C. The reaction was then stirred
at 50 C for 1.5
hour, and quenched with a saturated aqueous solution of sodium bicarbonate.
The resulting mixture
was extracted with dichloromethane. The combined organic extracts were washed
with 0.1 M
solution of hydrochloric acid and a saturated aqueous solution of sodium
bicarbonate, then dried
over anhydrous MgSO4 and concentrated in vacuo. The crude product was purified
by flash
chromatography (n-Hexane/Ethyl Acetate=5/1) to afford the title compound (70
mg).
1H NMR (400 MHz, CD30D-d4) 5 7.54 (s, 1H), 7.30 (s, 1H), 7.22 (t, J = 8.0 Hz,
2H), 7.00 (t, J = 8.0
Hz, 1H), 6.83 (d, J = 8.0 Hz, 2H), 3.81-3.43 (m, 8H), 3.08 (t, J = 7.2 Hz,
4H), 2.26-1.86 (m, 4H),
1.07-1.00 (m, 4H), 0.77 (1, J = 7.2 Hz, 6H). MS nt/z: 490 [M+1]*.
Example 60
3-(Dibutylamino)-N-(2-hydraxyethyl)-4-phenoxy-5-sulfamoylbenzamide
NTP-3033
4.0
VIA0,--1-
General Procedure N
OH
0 OH 0 IRII,)
H2N,91 0soc,2 y-' + H2N 11 DCM H2N.9, 40 N-----
...õ-----õ,
40 140
3-(Dibutylarnino)-4-phenoxy-5-sulfamoylbenzoic acid (100 mg, 0.24 mmol) and
dried
dichloromethane (3 mL) were placed in 25 mL round-bottom flask with a magnetic
stirring bar.
Thionyl chloride (0.05 mL, 0.713 mmol) was added drop wise into the above
solution at 0 . The
mixture was stirred at room temperature for 1 hour. The solvent and excess
thionyl chloride were
removed on a rotary evaporator. Dichloromethane (3 mL) and 2-aminoethanol (29
mg, 0.48 mmol)
were added to the residue at 0 C. The solution was kept stirring at room
temperature for 1 hour.
The solvent was removed and the crude product was purified by flash
chromatography (n-
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Hexane/Ethyl Acetate=5/1) to afford the title compound (40 mg). IHNMR (400
MHz, CD30D-d4) 6
8.02 (d, J=3.2 Hz, 1H), 7.75 (d, J=3.2 Hz, 1H), 7.24 (t, 3 = 8.0 Hz, 2H), 7.00
(t, J = 8.0 Hz, 1H),
6.83 (d, J = 8.0 Hz, 2H), 3.72 (t, 3=7.6 Hz, 2H), 3.52 (t, 3=8.0 Hz, 2H), 3.09
(t, J= 8.0 Hz, 4H), 1.27-
1.20 (m, 4H), 1.06-0.96 (m, 4H), 0.77 (t, 3 = 7.2 Hz, 6H). MS m/z: 464 [M+1]+.
Example 61
3-(Dibutylamino)-N-(1-hydroxypropan-2-y1)-4-phenoxy-5-sulfamoylbenzamide
NTP-3045
OH
H24 0 /-,,T--
General Method 0
OH
0 ON OH
0 [d)
H2N,,)
H2N,II 0= /
H2N.3 410 N/-1
8 0 HATU, DIEA, DMF
0 0
A reaction vial was charged with 3-dibutylamino-4-phenoxy-5-sulfamoyl-benzoic
acid (70 mg,
0.166 mmol), 2-amino-l-propanol (15 mg, 0.1999 mmol), 0-(7-azabenzotriazol-1-
y1)-N,N,N,N-
tetramethyl uranium hexafluorophosphate ( 76 mg, 0.199 mmol), N,N-diisopropyl
ethyl amine (33
uL, 0.199 mmol), DMF ( 2mL) and the mixture was stirred at room temperature
overnight. The
reaction was diluted with ethyl acetate and washed with water, brine and dried
over Na2SO4. The
solvent was removed under reduced pressure and the residue purified by flash
chromatography to
give the product as white solid (69 mg). IHNMR (300 MHz, DMS0- c15) 6 8.29 (d,
3=8.1 Hz, 1H),
7.94 (d, J=2.1 Hz, 1H), 7.67 (d, J=1.8 Hz, 1H), 7.25 (s, 2H), 7.20 (t, J = 7.2
Hz, 2H), 6.96 (t, J = 7.2
Hz, 1H), 6.73 (d, J = 8.1 Hz, 2H), 4.75 (t, 3=6.00 Hz, 1H), 4.06-4.4.01 (m,
1H), 3.51-3.22 (m, 6H),
3.00 (t, J= 7.2 Hz, 4H), 1.19-1.09 (m, 4H), 1.01-0.89 (m, 4H), 0.72 (t, J =
7.5 Hz, 6H). MS m/z:
478[M+1]+.
Following General Methods M through 0 or with slight modifications thereof,
and following
procedures familiar to one of ordinary skill in the art, the following
examples were prepared from
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benzoic acids which can be prepared following General Methods A through P and
commercially
available reagents.
Example NTP General Structure M.W. Mass Spec.
Number Number Method calculated found
miz
62 NTP-3001 M crN 0 495.63 496[M+1]+
H2N,
CI
63 NTP-3002 M 544.11 544
10 H
H2N.
" 0
64 NTP-3003 M 509.66 510[M-1-1]
la_11 0
000
0 '-
65 NTP-3005 M0 510.65 511[M-Fl]
0-0 0
66 NTP-3006 M 0 0 500.61 501[M+1]
H N..a 110
ro 0
67 NTP-3007 N 486.59 487[M+1]+
HNLL-7,-,
00 01[5 s\_
-
OF,
68 NTP-3008 N 563.63 564[114+1]-
Har-
2N0'0
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69 NTP-3009 M
F,C,cr0 563.63 564[M+1]
0.1:5--\\_
00
-
70 NTP-3010 N563.63 564[M+1]
H2N10 0 0.1:5_,\_
71 NTP-3011 M
* m,,r0 541.68 542[M-1-1]+
cr:,)
72 NTP-3012 M
IdH 0 541.68 542[M-F1]
F 0
Fi2N,
F
73 NTP-3013 M
545.64 546[M+1]+
.0
74 NTP-3014 ' M
578.55 578,580[M+2]+
coli)a-
H214,4N,--i¨
de 00 õ
CI H
75 NTP-3015 N
o.,,,,4, 530.08 530,532[M+2]+
14 11,s.
2 60 õdm_
76 NTP-3016 M
,Nõr.0 530.08 530,532[M+2]+
0-0
77 NTP-3017 M
farl; 0 523.69 524[M-F1]+
' 78 _ NTP-3018 Mc):),.: H _
564.52 564,566[M+2]
A
k)
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79 NTP-3019 N546.68 547[M+1]
80 NTP-3020 N:4
496.62 497[M+1]
00 0,%."---
1
81 NTP-3021 N496.62
497 [M+1 ]
Ni:N If -.: .--/-
82 ' NTP-3022 N a .
459.60 460 [M+1]+
0 01$...õ ---/-
,
83 NTP-3023 M a 0
473.63 474[M+1]
H24,,-/-
o"0 . .,- \ _
I.
84 NTP-3024 M487.65 a 0
488 [M+11+
0"0
IL
85 NTP-3025 M'N'Th 1,..N 0
502.67 503[M+1]
F42N44.--f".
cro 0...6-
' 86 NTP-3026 NCb.r.
541.75 542 [M+1 ]
,At.,,,,_/-
0.
87 NTP-3027 M
555.77 556[M+1]+
Cb.
0001,¨\_
88 NTP-3028 N )1-Z-kr.
528.71 529[M+1]
. o
89 NTP-3029 M v'' LA 0
542.73 543[M+1]
He4,a4, N,
d o 0,.0`--\_
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- 90 NTP-3030 N a,. Jr
516.7 517[M+1]`
....¨
91 NTP-3031 M... õ,,
546.72 547[M+1]
.,:$
00 0,0
92 NTP-3034 M
419.19 420[M+11+
"4
93 NTP-3035 M....1..:1,1
475.64 476[M+1]
Hz,, <¨
h0)
94 NTP-3036 N .). .
475.64 476[M+1]
142t4 te¨r-
0 0 '90-
95 NTP-3037 N
564.52 564,566[M+2]+
.2N:41e_r_
0,--6-
00 0,1:5--\_
96 NTP-3038 M A .-
530.68 531[M+1r
e iL:%:, 14 t 4
H .,--/¨
-,11 0
97 ' NTP-3039 N
532.70 533[M+ 1 ]
0 4
ii o
98 NTP-3040 N
433.56 434[M+11
)14
A ot<
nõ.4_/_ 447.59 448[M+1]
99 NTP-3041 N1
00
k;
Ef2N,....,
100 NTP-3042 ML.,,,
502.67 503[M+1]
A 0
Followed by
H N..s 110 N.---/¨
deprotection
of the boc-
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protected
primary
amine with
HC1
101 NTP-3043 Mx Ho:l 502.67 503 [M+1]+
Followed by " 0,0
deprotection
of the hoc-
protected
secondary
amine with
HC1
102 NTP-3044 0 0 544.75 545[M+1]
103 NTP-7001 0 405.51 406[M+1]+
rjr
104 NTP-7002 0 0,6 419.54 420[M+1]*
6"o W
105 NTP-7003 0 461.57 462 [M+1]+
di) 0
o
106 NTP-7004 0 01A, 447.55 448[M+1]+
0- 0
o
107 NTP-7005 0 o. 461.57 462[M+11+
N.
cro o
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108 NTP-7006 0 0
503.21 504 [M+ ir
00 .
109 NTP-7007 0 0 It
433.56 434[M+I]
19 0,(y
110 NTP-7008 0
489.63 490 [M+1 r
[I
00
111 NTP-7009 0
517.68 518[M+1]
112 NTP-7010 0
461.62 462 [M+1 ]
A 406 0
113 NTP-7011 0
475.64 476[M+1]
)1,2g
114 NTP-7012 0 oJ1
524.67 525 [M+1 r
8 ,a---
115 NTP-7013 0
537.71 538[M+1
oo
116 NTP-7014 0
333.83 334[M+1]+
0
Aci
144
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117 NTP-7015 0
0 NI 447.22
448[M+1]+
1 0 a
-.4r.0-
8 06
H
118 NTP-7016 N
0 N 475.60
476[M+1I+
H 102
.ri'l'S NI,
IN
119 NTP-7017 N
489.63 489.63
490[M+1]
P 1 1-P--V¨/¨
00
120 NTP-7018 M
531.67
532[M+11
. 0
! v? 01 ,-/-
!=
0 0
i -c)
121 NTP-7019 N
551.70
552[M+1]
: 0 [1 40
t44c?,I1, .¨/¨
'y -. N,
0 0 L-2-1 0,,_____
H
122 NTP-7020 N
01N,LrN 538.66 539[M+1]
123 NTP-7021 0
447.59 448[M+1]
+
.,...cJ5L1
..---]
124 ¨ NTP-7022 0
0 517.68
518[M+1]+
0 c--)2 0,,I..1.-- \*..
041H,
125 NTP-7023 0
335.38 336[M+1]+
H2N.9
.. [1
0 c¨) 0,,_.,
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126 NTP-7024 0r---.0 ..TN,1 405.47 406[M+1r
,Ni
,
O00
2
127 NTP-7025 0 335.38 336[M+1]
FI,N1 N/
8 or)
128 NTP-7026 0 0 405.47 406[M-F1]
HoLigme,
6 06
129 NTP-7027 0 yH 393.46 394[M-F1]
0 c--7 0,,...,
130 NTP-7028 0 ; ,, rj 375.87 376[M-F1r
N
o a
piyr
131 NTP-7029 0 363.86 364[M-F1]
Foy,---------
0 a
0 2
132 NTP-7030 0 391.48 392[M+1]
H2N4
0 c-/ 0,7r..-
NTP-7031 0 r):' 461.57 462[M+1]
133 0 11,.....,
H2N1 Ni"---7--
O ob¨
OH
134 NTP-7032 0 0 N,r) 449.56 450[M+1]
Fol
8 o,...
c---7
,.:13ilki,
135 NTP-7033 0 487.57 ' 488[M+1]+
FI,N1 .
0 Cx.613
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136 NTP-7034 0
557.67
558[M+1]+
H2N1 N * 0 0.60
137 NTP-7035
H2NS NO
0 0 * 6 NH2 375.45
376[M-1-1]
138 NTP-7036
HI 1111
mn0 N..õ. 445.57
446[M+1r
139 NTP-7037 0
H2Ni
06-/ Nr--\OH 433.52
434[M+11
140 NTP-7038
8 0 H N/---/--- 381.42
382[M+1]+
141 NTP-7039 00 OT..J
H,N.V
8 0 H OH 439.50
440[M+I]
142 NTP-7040 0
H2N1
1110 8 0 = NH2 449.56
450[M+1r
143 NTP-7041 0
H,N.1
41018 0 p0 NH2 433.56
434[M+1]
144 NTP-7042 0
503.55
504[M+1r
0..c7õ.
147
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145
NTP-7043 0
041,`"
491.64 492[M+1]
HA,Csi
õ---/---
047
146
NTP-7044 0
H,N
449.56 450[M+1]
4
.---7--
8
`0
147
NTP-7045 0r'o
0 N..,.)
519.65
520[M+1]
H2N .V 0 /---/-
Z--
0-
H CM
507.64
508[M+1r
-148
NTP-7046 0
z
0
C;
0--
149
NTP-7047 0
04N:LH,
488.43
488
H20
/-----7---
0
-...r*CI
CI
150
NTP-7048 0. r11,)OH--- 435.54
436[M+1]
õHy.
,
N----,
0 0,
o
4,,)H
419.49
420[M+1]
151
NTP-7049 0
H,N4 n
8 0.6--
0 NH,
152
NTP-7050 0
473.63 474[M+1]+
C1N,2 0 ....,......._
S
N
8
(-)
153
NTP-7051 0
ro
0 N....)
543.72
544[M+1]
ay.
0
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01-1
154 NTP-7052 0 0 NT) 531.71 532[114+1r
CIN4
0 NH
155 NTP-7053 0 447.59 448[M+11+
?iv
OH
156 NTP-7054 0 0 11,) 491.64 492[M+1]*
_O5
NTP-7055H OH
157 0 Nyi 505.67 506[M+1]+
o o
Example 158
3-Dibutylamino-5-hydroxymethy1-2-phenoxy-benzenesulfonamide
NTP-10001
OH
H2N4
o
General method P
0 OH = 0, 0 0
H2N3 III) soc,r H2N . 9
S N S N
0 0 Me0H 8 0 ACN 0 0
40
OH
LiAIH4 Fi_N3
THF S N
8 0
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Step 1: 3-Dibutylamino-4-phenoxy-5-sulfamoyl-benzoic acid methyl ester. A
round bottom flask
was charged with 3-dibutylamino-4-phenoxy-5-sulfamoyl benzoic acid (1.82 g,
4.33 mmol) and
methanol (50 mL). Thionyl chloride (L50 g, 12.99 mmol) was added slowly at
room temperature
and the reaction mixture was heated to 50 overnight. The solvent was removed
under reduced
pressure and the residue re-dissolved in ethyl acetate and washed with
saturated sodium bicarbonate
solution, water and brine. The organic solvent was removed under reduced
pressure and the residue
purified by flash chromatography to give the product as white solid (1.75g).
MS m/z: 435 [M+11*.
Step 2: 3-Dibutylamino-5-(dimethylaminomethylene-sulfamoy1)-4-phenoxy-benzoic
acid methyl
ester. A reaction flask was charged with 3-dibutylamino-4-phenoxy-5-sulfarnoyl
benzoic acid
methyl ester (1.75 g, 4.032 mmol), acetonitrile (25 mL) and N,N-dirriethyl
formamide dimethyl
acetal (0.56 mL, 4.233 mmol) and stirred at room temperature for 2 hours. The
solvent was removed
under reduced pressure and the resultant gummy residue was treated with ice
cold water to give pale
yellow solid. The solid was filtered and dried to give the product (2.1g). MS
miz: 490 [M-1-1]+.
Step 3: 3-Dibutylamino-5-hydroxymethy1-2-phenoxy-benzenesulfonamide (NTP-
10001). A round
bottom flask was charged with 3-dibutylamino-5-(dimethylaminomethylene-
stilfamoy1)-4-phenoxy-
benzoic acid methyl ester (200 mg, 0.41 mmol), THF (5 mL) and lithium aluminum
hydride (2M in
THF) ( 0.25 mL, 0.49 mmol) was added and the reaction stirred at room
temperature for 1 hour.
Another portion of lithium aluminum hydride (2M in THF) ( 0.25 mL, 0.49 mmol)
was added and
the reaction was heated to 50 C for 1 hour. The reaction was cooled and
quenched with cold water
and diluted with ether. The reaction was filtered and the filtrate removed
under reduced pressure.
The residue was then purified by flash chromatography to give the product as
white solid (33 mg).
1H NMR (300 MHz, DMSO-d6) 7.41 (d, J=1.8 Hz, 1H), 7.21-7.16 (m, 3H)7.13 (s,
2H), 6.93 (t,
J=7.2 Hz, 1H), 6.71 (d, J=8.7 Hz, 2H), 5.35 (t, J= 5.7 Hz, 1H), 4.51 (d, J.--
5.7 Hz, 2H), 2.97 (t, J=7.2
Hz, 4H), 1.17-1.1.10 (m, 4H), 1.02-0.94 (m, 4H), 0.73 (t, J=7.5 Hz, 6H). MS
m/z: 407 [M+1 ]+.
Example 159
3-(bis(3-chloro-4-fluorobenzypamino)-5-(hydroxymethyl)-2-
plienoxybenzenesulfonamide
NTP-10002
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OH
CI
H2NI 11101 N = F
(6'
General method Q
0 OH OH
CI CI
H2N,9 JON F SH&THF 1 H2N.9 N F
0 0 AI THF 0 0
40 uir F 40 10
CI CI
3-(bis(3-chloro-4-fluorobenzypamino)-5-(hydroxymethyl)-2-
phenoxybenzeriesulfonamide: A
reaction vial was charged with 3-(bis(3-chloro-4-fluorobenzyl)amino)- 4-
phenoxy-5-sulfamoyl-
benzoic acid (75 rug, 0.126 mmol), tetrahydrofuran (4 mL) and BH3.THF complex
(1.0M in
tetrahydrofuran) was added drowise. The reaction miture was stirred at room
temperature for 1
hour. Water was added and the reaction miture was extracted with ethyl
acetate, washed with brine
and dried over MgSO4. The solvents were removed under reduced pressure and the
residue purified
by flash chromatography to give the title product (48 mg). 11-1 NMR (300 MHz,
DMSO-d6) (5 7.52 (d,
J=2.1 Hz, 1H), 7.34 (t, J=7.5 Hz, 2H), 7.26 (s, 2H), 7.22-7.11 (m, 4H), 6.91
(dd, J=7.5 &2.1 Hz,
2H), 6.83-6.77 (m, 4H), 5.34 (t, J= 5.7 Hz, 1H), 4.44 (d, J=5.7 Hz, 2H), 4.11
(s, 4H). MS m/z: 580.9
[M+1].
Example 160
3-(butylamino)-5-(hydroxymethyl)-2-phenoxybenzenesulfonamide
NTP-10003
OH
1-121,141
. 0 H
=
The title compound is made according to general method P, beginning with
bumetanide in step 1.1H
NMR (300 MHz, DMSO-d6) 6 7.25-7.19 (m, 2H), 7.07 (bs, 2F1), 7.05 (d, J=2.1 Hz,
1H), 6.95 (t,
J=7.5 Hz, 1H), 6.86 (s, 1H), 6.81 (d, J=7.5 Hz, 2H), 5.28 (t, J= 5.7 Hz, 1H),
4.67 (t, J=5.7 Hz, 1H),
151
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4.48 (d, J=6.0 Hz, 2H), 3.04-2.97 (m. 2H), 1.37-1.30 (m, 2H), 1.14-1.06 (m,
2H), 0.76 (t, J=7.5 Hz,
3H). MS m/z: 351.0 [M+1].
Example 161
3-(dibutylamino)-N,N-diethyl-5-(hydroxymethyl)-2-phenoxybenzenesulfonatnide
NTP-10004
OH
0 1101 _
0 0 \
The title compound is made according to general procedure Q, beginning with
the appropriate
benzoic acid derivative. IHNMR (300 MHz, DMSO-d6) ö 7.42 (d, 1=1.2 Hz, 1H),
7.20 (t, J=7.8 Hz,
3H), 6.94 (t, J=7.5 Hz, 1H), 6.63 (d, J=7.8 Hz, 2H), 5.35 (t, .1= 5.7 Hz, 1H),
4.52 (d,1=6.0 Hz, 2H),
3.24-3.13 (m, 4H), 2.95 (t,1=7.2 Hz, 4H), 1.18-1.08 (m, 4H), 1.01-0.90 (m,
10H), 0.70 (t, 1=7.2 Hz,
6H). MS mi.z: 463.3 [M+1].
EXAMPLE 162
Additional Arylsulfonamide Compounds
[02663 Additional arylsulfonamide compounds may be synthesized using standard
methods.
Following General Methods A through P or with slight modifications thereof,
and following
procedures familiar to one of ordinary skill in the art, the following
examples can be prepared from
commercially available reagents.
152
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HO 0 HO 0
HO 0
HO 0 HO
0
H2N, , 0 N.--,õ/"...õ H2N .s .
N.., 0 H2 Ns , *
H N 0
le'"'''' 2 S, W
OH H2N;5 , 1111 N''""`
4 ',µ
04'sb 0 H 0 0 0 1,...1
cro 0 LI
0 0 0 i,t, cro
0 H
0
0 OH 0 OH
* ' 0 '
HO 0 HO 0
HO 0
HO 0 HO 0
I
HAI. s 5 , N H2 N ,s . N ------
.õ..,---, N.," H2 N5 0 re-,.0,... H2 N,s * N,----,õ...-----,, H2N , IP Nr."-
......"..,
; N".-N"" N
0 4,, 4,,
0"0 0 LI 000
I 0 0 I,,
0 0 0 1...1 0"0 0
1...1
0 ' '0 H
0
11111 (I)
SO
HO 0 HO 0
HO 0
HO 0
HO 0
r---- N -
F
H2N.c 1101 N..---.õ,---,õõ H2N8 , III N....õ...õ---,õ H21-J5 41)1 N"--,õ..-N
H2N, 1111 N.----,,0õ.õ...- H2N,s 110 N-----,./14
..A ON
Crb 0 0"O 0
in 0 LI
0"0 0 Li
0N 0 o 1
0 r..,11..1
N
0 0.1
5
c o) 0 F F
5 0 N
F HO 0 I
HO 0
HO 0
HO 0 ,1-10.
, 0
H2N H2N , 0
001
FI2 N . , 0 -----,- -0--,--
S N
N
H2 NS, 0 '---.
N ------' H2 N, 0,s , ,S, N
00 0 01'0 0
0 N
ci''''b o Li
0"0
" 0
411 01=0 (00 410
0 0
S.
0 I:) N.--.
F
H2N 0 H2N 0
1-12N 0
H2N 0
H2N 0
H2N.-' * N....--....õ---,... H2 NS
0N 2 H NS ......õ.õ... --,õ H2 N,
$ N ,, N
1111 N".....OH H2NL;5,
N
04 H0"0 0 1.õ1
CPO 0 LI
0"0 0
0"0 0
0 OH 0 OH
H2N 0 H2N 0
I-12N 0
H2N 0
H2N 0
I
, 0 N H N 0N..--,õ...-*,N,'
H2N , s 0 N
0 H2 NS 0 N,-,.õ. HAI , c * N....õ.........õ.
V'''. s '" 2 ',S ,
. ''
4e ,
0"0 0 H o"o 0 1.1, I
o , 0 o LI
cro 0 H
'`O 0 H
0 ,..N.,..,
0 0 OH SO *0
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H2N 0 H2N 0 H2N 0
H2N a
H2N 0
(-re' F
H2N, 0 N.,-..........----, H2N., 1101 N^...,--", H2N,is, 0 N.,-..õ,..N`--)
H2N, 0 N,..-..õ......0õ,,, H2N,A * ..".õ).
,S,
N F
t=-=+. 69,,
o"o 0 H o"o 0 1,,,Li< o"o 0 I.,1
o"o 0 I..,
o `o 0 ci
0 (N) F F N
0 01
0 F? F
10 F
* C j
CO) N
F i
I-12N 0 H2N 0 H2N 0
H2N 0
H2N 0 so
,....-
H2N., 1111 V^....... H2N.5 H2N N ..õ 110
N 1111 .
lip H2N,Q H2NS 0
e'N. N 1110 t- /-%
Cr 0 0"0 lo 00 0 000
000 1..n.,
0 I
0 0=r0 10 IS
F N.'
55 0 0
HO 0 HO 0 HO 0 HO 0
HO 0
H2N, 0 N
1-12N;S, 5 NC) 112NS, = N- H2NL5 . NO/-I H2N;S, 0
ono 0 1,1 6 µo 0 . L
0 I o"o o t...)
0 õ...N,....
OH OH
5
F CI F
F CI F
HO 0 HO 0
HO 0 HO 0 HO 0
I
H2N... .
r=-=,, A N).'''''N'" H2N-s 0 N.,-....õ0õ,õ H2N,5 0
N,"....õ--",õ H2N5, 0 N-",õ....".õ
, s 4,,
000 1.õ) 00 0 ci I
o"o 0 1.1 o o 0 H o"o 0 H
0 ,N,
0 OH
CI 41 10 (I)
SO
F 1 CI F
CI
F CI
HO 0 HO 0 110 0
HO 0
HO 0
F
rir ,,,,, õ......õ)<F
H2N,Q . N....-õ,...,-,,, H2N,A, 10 N"....."--, H2N,q 1110 .---..-,N---, ¶2-
,s 1111 1,10õ,, NALA 100
i=-,
N F
N .. \
i;-,.
0"o 0 Li 00 0 1,11( 0"0 0
ci 0 0 0 I..,
0 µ0 0 1.õ1
F F 0 cN,
5 F.---,F F
F
F s 01
F F
0 F
F N
CI
F CI I
F HO 0 HO 0
HO 0
HO 0
HO 0 s
'N
H2N 411 H2N5 . 110 110
H2 NS 110 N .q 1101
N H2N,, 110 9
'' ,-% ert..
1 N---" 112N., IP
0 0 0 N
N 0"O 0 i-,,
0"0 0 LI i-4.
0" 0
0"0 0
0 0=T=0
0 Cn
* 401 0 rc
-...0 F F 40 0
41 a0
F Cl
CI
......o
154
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,
HO 0 HO 0 HO 0
HO 0 HO 0
- ,S, N".....'" '''''.."M'A 0 N ^--- -- C1NL,s, = N'''''''. ."-
N.;S, N''''''OH
ot b 0 t...i
0"0 0 LI 0"0 0 1.1
00 0 1,1 boo Li
40
0 OH 110 OH
HO 0 HO 0 HO 0
HO 0 HO 0
HN., * N..--.õ...,,N., 1-1Nr: 0A 0 *
NI 0
010 Lli 1 NC'''
000 1õ,i 00 oLI
cro 0 LI o"o 0 Li
5 õ...N,õ
le O
., OH
* 0
SO
HO 0 HO 0 HO 0
HO 0 HO 0
0"Th 0
,-----N- F
r
1 f---
cõN N"..."''''''''. /NI'S (6 N',""=-= ''S, s N''''''N.'")
.....11, 0 ...".......,0õ....,., CIN,s I. r4.----,F
"A
A N
Cr0 0 Li 0 0 0 1.11< 0"O 0 LI
o"o 0 LI cm 0
0 cN) v F N
40 01 011 F? F
0 F I.1 0
0 N
HO 0 F HO 0 I
HO 0 HO 0
HO 0 .
'N
0 ,c, 40 "" 140 ill * Y
N, 0 N'''''''= EN1 , 1101N. 0 N
N - ,S, N
crA ' A 00 0
0"O 0 0"0 0
o"o 0 N
00 01=0
00 * 40 1401 40 001
5 N
F
HO HO HO
HO HO
H2N. 1101 N"--õ...õ..,...õ H2N,5 0 N.....",õ.0,.. H2N.5 5 r4,,......,,,..,.
H2N, 11101 Nr....õ..,,,OH H2N,,s,
/.r.:µ 4 µµ
eo 0 1,1 0 o 0 LI o o 0 LI
cro 0 LI o"o 0 LI
40 0, SO
0 N ,
0 OH OH
0
HO HO HO
HO HO
I
H2N.,s, 1101 N"---......-",N, Fl2N, * 0 H N 310
N.,--.,-...õ H2N, 110 N.--",..",..
.,S
N''' .."- 2 µ,S, ,S,
CPO 0 I eb 0 l, o"o 0 Li
o"o 0 Li
0 0i, OH SO
SO
HO HO HO
HO HO
H2N., lb N'''''''''''' H2NA N.".''- H2147/5% 5
WA."'") H2rl'e 16I N'''''-'" 1-121\1',,S, 5
eo o o"o 01,11< o"o 0 LI
crt 0 H ono 0 L.
I (N) 0 0 F ; * N
40 h 401 F"'-''FF
i
155
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F HO
HO
HO HO
HO
ri'll
ISI I-12N ,, 1101
* H2N ...is, 110
H2N A 10
N
N = H 0
N H2N,s 101
01% 0
OA) 0
2N,s 1., %.= N
00 0 L.1
0 0 0
OTh 0 N
0 01=0 "
* 0 001 401
0 40
N
F
HO HO 0
HO
HO 0
HO
I 0 H
N''' --.N A = N C,S,
N' ""N'A,
Nre.----"
0 FOH =-"N`.,S, N'-'"
O"O 0 1...11,1
0"0 0 LI,
O"O 0 Li,
1.1
0
0 OH 0 OH
F
CI
F
F CI
F
HO HO
HO 0
H2N 0
HO
r I
.........., * r , re",,,,N,..
1
A, NH * N."....õ--
-", .--' N. * ry\----11, -IA I. NCS, 0 N'''''-'''''
" A N ."-. A
o"o 0 1õ1 00 o
o"o o 1õ1,
o"o 0 1.1
o"o 0 1,1
so c.)
1#01 OH
CI 0 F I
0150
F
CI
F
CI
HO 0 HO 0
HO
HO
H2N 0
1 0 F
..- A k 101 r *
r.:)NrO s 01 tO .õ...-- ..,s,
-
---.....õ)< .
N.,-,..,..õ-----,:-,..õ. N., N'''' ..,
s., 1110 N-"\---N ....
N.'
N F
00 0 H cr'b 0
[.,11 F
or, .6 0 H
o"o o L.
0"0 0 ci
F 0 cNN)
401
F".."'FF
F I.
F 4111 (N)
F F
F* h
0 FF
F
CI 1
CI
F H2N 0
HO
H2N 0 HO 0 0
H2N 0
r?
,õ..."0 0õ10 N I--
k
40 a,/
si ,1 0 ji
.,õ.., ,0 0
- A * N
S * N
N..Q
0
0"0 0
0"0 0
N
0"0 0 N
0 (I= 0
5 LIC)
0 F F 0 0
0 *
N
o 4111
CI
F a
0
CI
HO 0 HO 0
1-10 0 HO 0
HO 0
H2N8 . 0 Nr.^....,.."..., H2N ....,s, 0 w."..÷. H 2 N . , 0 N....---,......,--
,õ H2Ns
, (110
H N *
.---....,..".õ
Kr.'"--" 2 S, Ni...)
o"o 0 cro c)
L.,
o4-, "o ci 1.1., cro 0
1õ o"o 0
F3c 0
0
* C F3 0
....., F 411 F
C Fa
F C F3
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HO 0 HO 0
HO 0
HO 0 HO 0
H2N, 101 tr.---,,,---,, H2N, 101 N.,--..õ....
H2Nõ . 110
N'-'---- H2N, 1110 N.-",õ...."--,õ 112N..i5, 11101
N........,
OA) 0 L 0"0 0 1....,L
Po 0 LL
o"o 0 1.1, o"o 0
a 0 .....õ
0 0 0
IS
0
Cl O-'
FiN /
0=S=0
1
HO 0 HO 0
HO 0
HO 0 HO 0
H2r1. 0 N--""''''''
H N 02 'S
, S, H2N;S, * N
I-12NA 11111
H2N. 0 A 1\' e0 01...
N--'
0A) 0 isi., 0"O 0
00 0
0"0 0
11, LI,
( 0
0 0 =-' N
I. 0
0
F i
*
H
HO 0 HO 0
HD 0 -FNI'=
HO 0 4111
HO 0
H2N,S ,,,,,...,,,, H2N7,s, IP
N----""-----%'- H2N,S , 5 N.--.õõ.....--..õ.
H 3N., 0
,, N F
"---- H2N , 10
0"0 0 0"0 0 (
0"0 0 LI,
ro 0 LI, ,s,
cOo o
/ 0
4111 n 0
LI.
0 N
i
H
H2N 140
CF3 F3C,.0
0
HO 0 HO 0
HO 0 HO 0
HO 0
i
fll, 101
sl'S I* N- as 0
101 ...--..õ....õ...-.õ.
NF."' N`Q N''
F. A N
(lb 0 00 0 [A,..,
04-'0 0 L.
04-'µµO 0 1.1.,õ 0 00
F3 C 410
0 0
C F3
4
0
F
C F3
F
HO 0 HO 0
HO 0
HO 0 HO 0
1 r-
a lill N-",,,-"". ...,N- 110 Kr"....-
^-õ N....4"N' 11111 N"---"--
A is,
, 110 N-'. ril
',S, 1111 ..----õ,N
000 LI, 0"0 0
0"0 0 1-1,
' A CPO 0
00 0
U 0
F aim LI,
L1-..
* 0 0
0
WI
CI 0--/
HN /
0=S=0
1
HO 0
HO 0 HO 0
HO 0
HO 0
0-Th 10
N.^...õõ...--,õ. ,,N=, 10 Ne"..
N, 110 ,....,õ. "'is,
0"0 0
0"0 0 0r1 0 l,
el 0
r A o
0 0 N411 0
140 it; 0
F i
H411likF
N
4111
..., ,....
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HO 0
HO 0
HO 0
HO 0
HO 0
111, 0
r+I
A
-"
iV, 0
N
I
N - -
1'S,
, HN.õ 0
0 0
--- N'S
N--"'"'' ¨ *
F' A
,
N
0"0 0
0"0 0
0'0 0 1,..1.,
,, N,
0 0 0
1...,L,
0"O 0I ,.
0 0
0
*
/ a
H
H2N 0
eF3
F3C.0
0
HO
HO
HO
HO
HO
I =
I
-A'A. N, a, so
.--õ, 0
N,c..N.s
1
N,...õ. .-N-,.,* N''''''''''
CrO 0
0"0 0
Oirb 0 1..
0 µ0 0
LIN.
Irr0 0
S CF 3
F3C 0
0 0
CF 3
F
F
HO
HO
HO
HO
HO
ON,S *
A
r--
N..--...õ,...õ--,,
NI , 0 N....---,--.õ, ',õ_õ.-N.s 0
''' N......"..
S * N.,,,, _.N5,_.N5oo 0
0
""
,0
0"0 0
11.
0 0 o
11...
cro o
eb 0
F ill& LI,
Cr 0
LI\
0
0
0
111V
o 2
GI
HN/
0,-S=.0
I
HO
HO
HO
HO
HO
0"Th Ili
I
H
,s 0 W.---.."--.
,s, 411111*104 N.-----õõ,-..õ.
,.....N,is, * N....--õ........,-...õ
...-NA 0 N-^-..õ...-^.õ
N,
0
........õ.õ---...õ 0 N,,,,.
O"O 0 [
0"0 0
00 0 L.
r A 0 T,t,
0 o o
LI.
0
00 :
N
0 o
0 0
F
I
H
0
0
HO
HO
HO
....,õ.HO
HO
IL 0
I
N.,--...,......--.õ.
...--NIs * ,
N, HN...is, = N.,-.õ.....----.õ. a, 0
I
NN
,S,N''''''''''''=
0"O 0
o"o 0
L1-..
o' '0 0
o"o 0
* 0
/
INL.
o"o 0 la
N ..."
HAI 0
CF3
FIC,.0
H
0
HO 0
HO 0
HO 0
HO 0
HO 0
IL 0
r 0
0
µr-."---"'"=
A 1111 NDH rd\l',S, = N=""'-'""
..-= ,s,
o' b aim LI
d I)
H0'0 ahn
0"0 arab,
1 0"0.4,... 1õ,
IIIV 0õ
141 0,
(Pi 1,1
F 111W l'IOH
Ll'OH
F
CI
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HO 0 HO 0 HO 0
HO 0 HO 0
I I r
1 0
N, 1. N,---õ,...N,... HtiNI,c 0
Nt.."-"The HN' * N"--""--`11--- CIN'S,l''''."--
"'NA
.5,-:, of A b
CP 0 iii,r&i 1,1 0 0 aim I
0 0
IIIVI N IIV
F 1.1 11--
-OH
CF3
HO 0 HO 0 HO 0
HO 0 HO 0
CrTh ii,
1
I.õ,,,N;S, 4111111.
N"...-- 0 rj. 01 N.-",,..-0,.../ CiN's .1 N;
N-----'"--- ,;,,,c,
". iSN deb
00 arab, 1., 0 0 Am CPO ahh, 1-.1
0.'0 Li 1--..
tip i!i w LI-,..K RP N
11111 01
Or
5 F"--'"F
F
F Co) F F C )N
OF3
HO 0 F HO 0 I
HO 0 HO 0
HO 0 0
N IOL ( 1.I , 140
k
N
' A /SA v,,
s,110 N,
N 00 " ail
-
O"O101
0"0
111, 0=s=0 Iii
111,
CI N
I 140 0
F 40 CI 0
F
HO 0 HO 0 HO 0
HO 0 HO 0
R2N. 1110
Is , A't.S,A) = N"----`-------" ."'" NA
N"OH /5 = N''.
(r0 0 db 0 O 0 LI,
0"0 0 LI, I 00 0 H
N
50K * OH
0 0
HO 0 HO 0 HO 0
HO 0 HO 0
I I r
MI ., 0 N.-^....õ..-^, N., HN,s 0101 N...---,..õ0õ 0. 0
1 11101 .
NstS , NF.
0"0 0 H CPO 0 1,,, I 60 0
LI, 0-0 0 1,1 d'o 0 H
40 OH
40
0 \¨/
HO 0 HO 0 HO 0
HO 0
HO 0
0-Th III0
F
N"--"- '''S (II N-----"'"Aµ4") NH 0
, , ' 'A
N------ ------ CIN-s I. NF
CPO 0 1..) 0 0 0 'b0 LI
0"0 0 LI CPO 0 L.,..
N ostAIF N
F
0 F? F
1411 C ) 0= Co)
N lei
F HO 0 I HO 0
HO 0
HO 0
HO 0 0
1 .2......N
til 019 N''''''' Ell, 0 ",s ,
14VX lel ,S, N N, )4 0 401 H 0 ',8, N
(PO 0 ,S, CPO 0
00 0 0"0 0
0"0 0 N
* 0=S=0
0 N
I
55
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EXAMPLE 163
Benzylic Alcohol and Amine Compounds
[0267] Additional arylsulfonarnide compounds may be synthesized using standard
methods. R1_8 are
as defined herein. In addition to the above examples, compounds of the
formula:
Fp, R2
R7
...A IP . R4
R8 'IS\ N
0"0 Rs 133
can be prepared following General Method Q or with slight modifications
thereof, and following
procedures familiar to one of ordinary skill in the art, the following
examples can be prepared from
commercially available reagents.
General Method Q
OH
OMs Fill
Z, ,
MsCI FIZR1 R2
n2
R 7 IN R7
A
_.AS 0 ,R4 base
7
R8
.õ..rsl, base R 'I\
d 'ID R3 R3 Y solvent R 8 IS \
Y solvent
R4\
R8õõ 1 'IS
11ro R, R3
Ob R8 R3
(A) (13)
[0268] Primary alcohols (A) of the invention which can be made starting from
benzoic acids of the
invention following method P or with slight modifications thereof, can be
converted to compounds
with a suitable leaving group such as a mesylate, tosylate or triflate using
procedures familiar to one
of ordinary skill in the art such as reacting the primary alcohol with
methanesulfonyl chloride in the
presence of The converted alcohols can then be reacted with suitable alcohols
or primary or
secondary amines to provide in the presence of a suitable base such as
triethylamine or
diisopropylethylamine in a suitable solvent such as dichloromethane or N,N-
dimethylformamide to
provide compounds of the general formula.
[0269] Alternatively, compounds of the general formula:
Ri
1
Nõ R2
R7
R8 A N
db R, k
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can be prepared following General Method R or with slight modifications
thereof, and following
procedures familiar to one of ordinary skill in the art.
General Method R
Ri Ri
I I
0 N,0 N,R2
1 12
BH3.THF
1.-
R7 R7
THF
A R8,..A 1111 .R4
R8 'S N
N
60 R5 1113 00R5 1133
[0270] Amide compounds of the invention which can be prepared following
Methods M through 0
or with slight modifications thereof, and following procedures familiar to one
of ordinary skill in the
art can be converted to primary and secondary amines by reaction with a
suitable reducing agent
such as borane-tetrahydrofuran in a suitable solvent such as tetrahydrofuran.
[0271] Following General Methods Q through R or with slight modifications
thereof, and following
procedures familiar to one of ordinary skill in the art, the following
examples can be prepared from
commercially available reagents.
o 0
I-I2N,Is SO Nõ
2N,sµ 41$ N..---...õ..-..,õ H2N;5 µ 10 N'
Hõ,..., 40 N ".....--",.. H,
'-
01'0 0 A 0
00 0 Cl b 0
40
so
SI 0
F
FF.. i?0 &N,Ici
I. 0 101 a
H2N, IP N
H2N.Is \ 40 N ,S\
.A 10 H2N , 1011
N.- ,Sµ
O"O 0
CPO 0
0"0 0 00 0 L,
0 ,
0
0 0
H H
H H
. .
1 0,)
H2N., Iii0 Nr^..,.---- --, H2N.A
II 110 ,,,,---,õ,.--N""H2N ,`, 1111 N.--,,,,^., H2 N .A lb N
/,
1 S-- b 0 o"o 0
o o" 0o "o 0
0 0 0 0
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H
A.,14
0 N
. 14
F
F,1
F'-'-'-'rj
H2N,c 1111 N-
H2N8
, 0
''
H2N., 0 NIF^..,---,., H2NS
, 0
I
N.
"
O'-'6 0
o"o 0
04% o
0 0 0
)
r---
1
H
er,
1
0,)
H2N, 1101 N.....õ,,,,,,,...
H2N, 0
H N 0
H N, 0
IS\
N
2 'iS \
N.
A
0"0 0
0"O 0
0"0 0 1
0 I
0 N
0
0 N
LN
ON
N"
H2N, c 0
H 2N '1.S *
N-
N
0*1 0
,S,
0//'''µO 0
000
0"0 0
0 '
N
HO õ_\
I
N
a
\......h
.....
N
0
H2Nõ5 I. N.^,",
H 2N 5 N2N, 01
,,sµ
NIF^..'"
NF''''"
H2N,q 110 N
,Sµ
011% 0
00 0
00 0
A o
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o
(.0
.....,N.....,..õ0
r-N--0
...
1
I
0,)
....,111...s 0 N.,õ¨...... 0. 0 N, H2N,,s,
il
0 N----------, --
0 N
IS,
011-b 0
0"0 0
0"O 0
00 0
0 '
0
0
F
CI 0
CI
0 0
50
F
o
F."- --"---.
0
N.
N.,--..,õ,......,..
H2N8
N
. 0 IT----..õõ...-\
-"- 'IS,
" 0 '--
---.N::sµ 4111112---IPP
N ,S\ "W.
"
0"0 0 L,
0"O 0
0 0 0
0 0
CI
F
CI
F
H
H
H
H
N
r, N
....,N...--..õ...,,,N
r-N-N
...
1
1
0,)
Ii
--- N is,
0"0 0
00 0
01 b 0
0"0 0
0
Si0
0
CI
F
I
r
I
H
N
r...N
.....
1
1
0,)
0, 0N
,
rt, $
-----,-",,,,
-...,,,
N------"-.
_N8
H
0
N.
IS1101
\
00 0
0"0 0
00 0
0"O 0
0
F
Cl 0
=...,,
0
=
F3C
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H
H
H
H
N
IN
NI
0)
I 110
0
N
H
.
õ....õ,.. ---, ......,s, 0 N.---,õ0,,
I
"'NA I. NOH
/,µ
0"0 0
0 'ID 0
01'0 0
cro 0
ial a,
F 00
CI
I. 0
1 0
I. OH
......'111..
I
r
I
H
N
=-, ---,õ....,.N
N
I
NI
(21)
N
õ '-s
1\ s
'''
NF(:)
S
H 0
k 0
H
N' . NOH
"
'
ik
0"0 0
0"0 0
N
0"0 0 H
0"0 0
0
CI
OH
F
F3C
Example 164
4-chloro-24dibutylamino)-5-sulfamoylbenzoic acid
NTP-9001
9
H2N-S
OH
8
Cl 10 N"--'-'"--'-'=
General Method S
0
0
HN/----7---
0
0
II
ii
, S
\--\\__
S
H2N 8 . OH
H2N, 8 0 OH
v.
/----/---
CI
CI
TEA, DME, 150 C, 4h, MW
CI
[0272] A microwave vial was charged with 2,4-dichloro-5-sulfamoy1-benzoic acid
(200 mg, 0.741
mmol), triethylamine ( 0.4 mL), dibutyl amine (341 mg, 1.85 mmol) and
dimethoxy ethane (2 mL)
and the reaction mixture was heated in a microwave reactor at 150 for 4
hours. The reaction was
cooled to room temperature, diluted with ethyl acetate and washed with water
and brine. The
organic solvents were removed under reduced pressure and the residue purified
by flash
chromatography to give the product as white solid (120 mg). 1H NMR (300 MHz,
DMSO-d6):45
8.18 (s, 1H), 7.49 (s, 2H), 7.42 (s,1H), 3.22 (t, J=7.5 Hz, 4H), 1.44¨ 1.38
(m, 4H), 1.27-1.18 (m,
4H), 0.83 (t, J=7.2 Hz, 6H). MS /viz : 363 (M+1)+ .
164
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[0273] Following General Methods S or with slight modifications thereof, and
following procedures
familiar to one of ordinary skill in the art, the following examples were
prepared from commercially
available reagents.
TABLE 4
Example Number: NTP Number .1 Structure
M.W. r.Mass Spec. found.(m/z) !
.1 . . . calculated I
165 NTP-9002 9 6
320.75
HpN-gOH
CI reTh
166 NTP-9003 9 0
278.71 279[MA-1r
H2N¨g OH
CI 111111).
0
167 NTP-90040 H2N-g
308.74 309[M+1]CI
8 go oH
0
168 NTP-9005
333.79 334[M+1]+
Fo-g dm
ci 4"r¨ N^1
169 NTP-9006 H2N^S9 0
318.78 319[M+1]+
8 OH
CI til3
170 NTP-9007 9 0
373.86 3741114+11+
0
In addition the above examples, compounds of the formula:
Ri
R2¨Z 0
R3
r-s4
=R7
R8
µ0 R5
where R5 is aryloxy, can be prepared following General Method T or with slight
modifications
thereof, and following procedures familiar to one of ordinary skill in the
art, the following examples
can be prepared from commercially available reagents.
165
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General Method T
OH R1
R1
I
1
2¨Z 0
R2¨Z 0
R3
R3
40 k.,,
R rJR4
R7 a , 7.
R 7 40 114
0 ,A, Base
R8 IS \
Solvent 0"o o
0/ µ0 Cl
(A)
411111
[0274] Aromatic chlorides (A) which can be prepared following General Method U
or with slight
modifications thereof, and following procedures familiar to one of ordinary
skill in the art can
further be reacted with substituted or unsubstituted aromatic phenols in the
presence of a base such
as potassium carbonate or sodium hydride in a suitable solvent such as water
or N,N-
dimethylformamide to produce compounds of the formula. Following General
Methods S and T the
following compounds can be prepared from commercially available reagents.
HO 0 if- HO 0 ri HO 0 if-
HO 0 ri HO 0
40 N.,.....õ-^,,,..., 0 N..õ..........,.., 0
N.,.....,-,...õ...- riki N õ,_...".õ,- 0
H2N,q H21\1,c H2N,
H2Ns, RIPP Hp.,
Orb 0 Orsb 0 000
000
40 00 40 F
40 00 .1
F ,..-- CF3
CI
----
HO 0 rr HO 0 r HO 0 r
HO 0 if HO 0 rf-
di, N-...."..---- 40 N,õ..õ,.--,,......,
N,......õ-...._,- di Nõ..õ....,--. dist. N .õ...."--
õ.....õ-
H2N, ,S, 41111" H2N, H2N, ,S,
tilir H2N, ,S, II" H2N,s 41111
000 Oirb 0 0' '0 0
0"0 0 0 0 0
00 CI
\
110 40
crcF3 0 N
JD 140
H
O_/ 0.....
OH
HO 0 _,---,_r 0 HO 0 ,---, f N.-- HO 0
HO 0 1õ) HO 0 rf
ilk N.õ..) di N) isli N
Aii. N,....õ---,0.--= di N.õ..õ,.....õ.0H
H2N.;s, up H2N, 111*1 H2N.,
RIP H2N-,s, 4111." Hp, ,s, 1111111"
cro 0 cro 0
cro 0 of No 0
000 11.
CI
\
0
010 0 1110 0-
CF3 SO N H 1.
0---/ 10
166
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[0275] Beginning with such examples and following General Methods L, M through
0, P and S, or
with slight modifications thereof, and following procedures familiar to one of
ordinary skill in the
art, the following examples can be prepared from commercially available
reagents.
---
..--
HO 0 r HO 0 (.1
HO 0 ri HO 0 r
HO 0 ri
, tillir , aivi N.......õ--,,..."
ill N,c, , so N.õ,.......õ...,
Ali N--....."-----'
H
I
I
N ......N õ,s,
N,
\--:
/S%
A
di,, %No 0
0"0 0 00 0
CIO 0
00 0
lei SI
411 40
SO
F
CI
CF3 CI
F ,--.
../
HO 0 ri HO 0 r
HO 0 rr
HO 0 r
HO 0 ri
rii N,........---,..õ---6, , 0 N,..........õ,......... ..._,N1,
Ai Nõõ..,..",õõ..,
H
H
N,
-.......õ.N,
11111-ki" ,S, 1111}111
ro 0 cro 0
000
db o
0"o o
40 CI
\
410 410
411 N
H
0--i 0 \
0,-.
HO 0 ,...--- HO 0 ......,HO 0 101
HO 0 r)
HO 0 (1
1-0
. N....)
Atli N.õ...."--, ....,
iiik. N 0 r 0
1
i
N, 1 0 N
HN, IP
r S a, ir
HN.,
IIIP
A,S,
00 0 I e0 o
0"0 0
(Po 0
o'so 0 I.1
* CI
\
00 40
0.,..CF3 411 N
IS
H
0_10
..,"
I rf
Hif
HO if
HO ri ..., 0 (...
....N
.--
ilkNõ,.......--,...õ.., 0 , ill N.õ.,.., \ ...../
N,......., \ ., Ai N=,..,"\-..-2 , fdI 1
H N,õ.....--..../ i
......N.;s, Mr
cro 0
0A) 0 0"A 0 0
cr0 o
00 o
0 411
00 40
SF
CI
F
CF3 ...-- CI
0-Th
(....'
R2N 0 [......., il . r
FIJ
HO if ...-
HO il
N..õ--..õ.0
N --,...õ \õ...,-- 1 0
r H 0 N
H ,
N
is, tillr
O'-`6 0 o"o 0
0"0 o
dip 0
0"o 0
0 CI
\
00/ 001
0,CF3 0 N
ill
0
H
0õ__/
167
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---
t)
I
H2N 0 if- LN 0 r
HO ri ,N
HO ri
Ail, 46 N......."...õõ
III 0 N....õ--,....."-
N..,,,,,,
r 0 Nõ..........õ...- H
0 H
A 0
00 0000 0 0"0 0
6'b0
0 CI
\
S 0
* N
so
a'ICF3
H
0-i 0
H2N 0 ro HO
..--.., 0 HO ri
r N"--- ,Ni
..N....,0 . r
....õ 0 N...) Aka N
iik, N.,..õ--,,o,- H
r. .
1
I
* N)
ON,Q 'pi HN, WI
,s, HN,
,s, w
000 0 0 0
O'-"o 0 *
000 o"o 0
* CI
\
0 0
110 0 N
0
o.,..0 F3
11
Example 171
Formulations for CNS-Targeted Drugs
[0276] Oral Preparations. The compounds as described herein may be formulated
for oral
administration. Exemplary oral preparations comprise a compound described
herein in the range of
about 10-60 mg of drug substance together with various inactive ingredients
such as
microcrystalline cellulose and other excipients, contained in a gelatin
capsule. Alternatively, the
active drug substance may be provided in tablet form, including about 10-60 mg
of drug substance
with microcrystalline cellulose, hydroxypropyl cellulose, magnesium stearate
and other excipients.
[0277] Additionally, for oral administration, the compounds described herein
may be used in the
range of about 10-100 mg/kg, together with various inactive ingredients such
as microcrystalline
cellulose and other excipients, contained in a gelatin capsule. Alternatively,
the compounds
described herein may be provided in tablet form, including about 10-100 mg/kg
with
microcrystalline cellulose, hydroxypropyl cellulose, magnesium stearate and
other excipients.
[0278] Intravenous Preparations. The compounds as described herein may be
formulated for
intravenous administration. In an exemplary intravenous formulation, each
milliliter of sterile
solution can include about 1-25 mg of a compound as described herein with
about 20-40%
propylene glycol, about 0-10% ethyl alcohol, optionally water, buffers (e.g.,
about 5% sodium
benzoate and benzoic acid as buffers), and preservatives (e.g., about 1.5%
benzyl alcohol as a
preservative.)
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[0279] Also, for intravenous administration, each milliliter of sterile
solution can include about 1-25
mg/kg of a compound as described herein formulated with about 20-40% propylene
glycol, about 0-
10% ethyl alcohol, optionally water, buffers (e.g., about 5% sodium benzoate
and benzoic acid as
buffers) and preservatives (e.g., about 1.5% benzyl alcohol as a
preservative.)
Example 172
In Vitro Pharmacology: NaliCT1- Cotransporter Assay
[0280] The effects of selected compounds were assessed for their effect on the
Na-4-1C-C1-
Cotransporter in vitro. See Gamba (2005) Physiol. Rev. 85: 423-493.
[0281] Material. The Normal Human Dermal Fibroblasts (NHDF), Fibroblast Basal
Medium
(FBM), FGM-2 bullet kit were purchased from LONZA (C-2511). For the
experiments the 3
following buffers were made: Buffer A: 5 mM KCI, 0.8 mM MgSO4, 5 mM Glucose,
25 mM
HEPES/TRIS pH 7,4. Buffer B: Buffer A with 127, 3 mM Choline Chloride, 1.63 mM
CaCl2, 0.9
mM ouabaine (Sigma, 03125) and lng/rnl bFGF. Buffer C: Buffer A with 140 mM
NaCI, 1.8 mM
CaC12, 1 mM Ouabaine, bFGF I ng/mL and 5 mEi/mL [86Rb] (Perkin Elmer, NEZ072).
[0282] Method. After 2 weeks of cell culture in the reconstituted cell culture
medium (FBM with
the FGM-2 bullet kit), cells were seeded in white 96 well cell culture plates
at 3000 cells per well
and incubated overnight at 37 C (5% CO2). The following day, the cell culture
medium was
replaced by a medium with 0.2% FCS (low serum) and incubated for 48h at 37 C
(5% CO2). Then,
bFGF was added in each well at a final concentration of I ng/mL and cells were
incubated 1 hour at
37 C (5% CO2). The cells were then washed in buffer B for 15 minutes at 37 C.
The buffer B was
then replaced by compounds diluted in Buffer C. After an incubation of 20
minutes at 37 C, each
well was washed with a cold MgC12 (0.1M) solution. Subsequently, 40ttL of
scintillation fluid
(Microscint-40, Canberra Packard) was added to each well. Plates were sealed
and incubated
overnight at RT. The radioactivity was counted in a 96-well plate counter. The
uptake was
measured as the percentage of Bumetanide-sensitive 86Rb influx. All
experiments were performed in
duplicate. See also Chassande, et al. (1988) Eur. J. Biochem. 171: 425-433.
[0283] Bumetanide (30 ttM) was used a positive control compound to establish a
specific activity.
The results were expressed as a percent of control specific activity
[100X(86Rb influx with
compound-86Rb influx with bumetanide)/86Rb influx in Buffer C-86Rb influx with
bumetanide] and
as a percent inhibition of control specific activity [100-(precent/control
specific activity)] obtained
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in the presence of the compounds listed herein in Table 18. These results were
expressed as %
inhibition at 10 M.
TABLE 5
Compound % of Control Value - % Inhibition of Control Values
3001 124.6 none
3002 85.3 14.7
3003 95.6 4.4
3005 - 107.1 none
3006 98.9 1.1
3007 60.7 39.3
3008 96.5 3.5
3009 121.8 none
3010 120.3 none
3011 115.5 none
3012 95.4 4.6
3013 115.1 none
3014 125.2 none
3015 107.7 none
3016 117.6 none
3017 132.8 none
3018 131.3 none
3019 71.3 28.7
3020 122.4 none
3021 110.5 none
3022 150.5 none
3023 133.1 none
3024 47 53
3025 82.5 17.5
3026 101.4 none
3027 76.7 23.3
3028 79.5 20.5
3029 97.1 2.9
4002 75.5 24.5
3032 68.2 31.8
3033 90 10
3034 99.4 0.6
3035 93.1 6.9
3036 85.5 14.5
3037 103.9 none
3038 96.8 3.2
3039 108.7 none
3040 66.4 33.6
3041 71.2 = 28.8
3042 110.9 none
3043 102.6 none
4001 104.6 none
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6001 115.7 none
6002 84.8 15.2
7001 115.7 none
7002 99.6 0.4
7003 64.6 35.4
7004 102.6 none
7005 117.9 none
7006 = 112.4 none
[0284] Compounds showing "% Inhibition of Control Values" between 20 and 50%
were indicative
of weak to moderate inhibition of NKCC activity. Compounds showing "%
Inhibition of Control
Values" lower than 20% were indicative of no significant difference between
the compound as
described herein and vehicle control values. Many of the compounds show little
or no inhibition of
NKCC, while some show moderate inhibition. None of these compounds would be
expected to have
diuretic activity comparable to bumetanide.
Example 173
Testing Compounds for Diuretic Effects including cumulative
urine volume, sodium excretion, and potassium excretion
[0285] Purpose. Renal function assessments, including cumulative urine volume,
sodium excretion,
and potassium excretion is measured in animals administered compounds, and
vehicle controls over
6 hours post-dose.
[0286] Methods. The animals are fasted overnight prior to dosing without
access to drinking water
prior to any pretreatment. Food and water are withheld until after the
terminal sample collection or
for the first 6 hours of blood sample collection, where applicable. Prior to
test compound
administration, all animals are pretreated with a single IP dose of PBx. At
approximately 5 to 6
minutes prior to dosing, all animals may receive a single oral (PO) gavage
dose of 0.9% Sodium
Chloride for Injection, USP, at a dose volume of 15 mL/kg. The vehicle,
diuretic (control) [(e.g.,
bumetanide, furosemide, piretanide, azosemide, and torsernide)], DMSO, and the
test compounds are
administered via a single IP dose at a dose volume of 1 mL/kg. Cumulative
urine volume, sodium
excretion, and potassium excretion may be measured.
[0287] Test compounds showing less cumulative urine volume, sodium excretion,
and/or potassium
excretion in comparison to a diuretic control may show less diuretic side
effects when used
clinically.
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Example 174
In vitro Molecular Tests of Select Bumetanide Derivatives on GABAA Receptor
Isoforms
Experimental design for selectivity screen
[0288] The addition of GABA to GABAergic cells activates the recombinant
expressed GABAA
receptors, creating an ion movement through the ion channel in the GABAA
receptor. The electrical
current generated by the movement of chloride ions into the cells can be
quantified.
[0289] Experiments are conducted examining the activity of a4-subunit
containing GABAA
receptors activated by 10 j_tM GABA in the presence and absence of 10 i.tM
test compound. HEK-
293T cells are transiently transfected with rat or human GABAA receptor
subunits. Whole-cell patch
clamp recording is performed at -50 mV unless otherwise indicated. Test
compounds are diluted
from a freshly made stock in DMSO, GABA is prepared from a frozen stock. For
each experiment
GABA or GABA + a test compound is applied for 5 seconds, and the electrical
current generated by
the movement of chloride ions into the cells are measured and recorded as a
trace of current versus
time.
[0290] In FIGURES 4-8, 100% response to GABA denotes no inhibition of GABAA
receptors. Any
value below 100% was indicative of inhibition of GABAA receptors by a test
compound.
Compounds described herein were tested at 10 KM GABA concentrations against
al, a4, ot5, and 0E6
GABAA receptor isoforms. Many of the compounds described herein show
inhibition of a4, oc5, and
NGABAA receptor activity but not al GABAA receptor activity. Therefore
compounds described
herein show activity as selective c(4-subunit containing GABAA receptor
inhibitors.
Classic GABAergic drugs
[0291] Classic GABAergic drugs (e.g., benzodiazepines) are non-selective
agonists that increase
both the amplitude and time course of inhibitory currents in GABAA receptor.
As shown in
FIGURE 2, at (A) FRISIUM (clobazam) an anticonvulsant, (B) AMBIEN (zolpidem) a
sleep aid,
and (C) VALIUM (diazepam) an anxiolytic drug, all increase both the amplitude
and time course of
inhibitory currents in GABAA receptor. GABAA receptor agonists activate GABAA
receptors at low
GABA concentrations, and while effective, also induce CNS side effects
including sedation,
decreased respiration, decreased cognition, and impaired motor function.
Acylsulfonamide Derivatives
[0292] Compounds described herein were tested at 10 KM GABA concentrations
against multiple
GABAA receptor isoforms (e.g., a1133y2, (74133'Y2, a5133y2, a613372)= Several
of compounds described
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herein selectively antagonize the a4GABAA receptor isoform by inhibiting
currents in the a4
GABAA receptor isoform. Several of the compounds may act as noncompetitive
inhibitors.
Compounds described herein may inhibit the currents in the parasynaptic a.
GABAA receptor
isoform in GABAergic interneurons. Compounds described herein may inhibit
a4GABAA receptors
pre-synaptically. Further, preferred compounds described herein will not
active al GABAA
receptors post-synaptically.
a1l3372GABAA receptor isoform
[0293] The al subunit is the predominant a subunit in GABAA receptors in the
adult brain. The al-
containing receptors showed no significant activation in response many of the
compounds described
herein. In general, the amplitude of the current associated with GABA
activation of these receptors
is not affected by preferred compounds, with a mixture of increased decay time
and decreased
amplitude seen at the highest concentration (1011M) but no alteration at lower
concentrations. This
is in contrast to the significantly increased positive modulation seen with
action of benzodiazepines
and other classic GABA-ergic agents.
aliadadas GABAA receptor isoforms
[0294] In cells transfected with GABAA receptor isoforms containing a5 subunit
subtype, 101.1.M
NTP-3032, NTP-3033, NTP-3034, NTP-6002, and NTP-6008 inhibited the receptor
compared to the
control condition in the prescence of 101.tM GABA alone. The results are shown
in FIGURE 3.
0C4/a6 GABAA receptor isoforms
[0295] Receptors containing the a4 and a6 subunits are inhibited by NTP-3032,
NTP-3033, NTP-
3034, NTP-6002, and NTP-6008 at 1011114. These results are shown in Figure 3.
These compounds
demonstrated marked inhibition of these receptor isoforms compared to the
control condition in the
presence of 101.1.M GABA alone. Additionally, these compounds demonstrated
little effect at the
post-synaptic al-containing GABAA receptors under the same experimental
conditions.
[0296] Several of the compounds described herein (e.g., 3032, 3033, 3034,
4012, 4015, 5007, 5008,
5009, 5010, 5011, 5013, 5016, 6001, 6002,6006, 6008, 6009, 6011, 6012, 7021,
and 10001) show
selective inhibition on a4 and a6 GABAA receptor isoforms.
[0297] Compounds described herein appear to have a different mechanism of
action upon the
GABAA receptors than traditional GABA agonist drugs. The primary effect is an
inhibition of
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chloride current reducing total GABA "drive." It is seen best at a4. and a6,
a5 and the inhibition is
consistent with noncompetitive antagonism, i.e., open channel block.
[0298] The activity of compounds described herein may exhibit two
features¨inhibition of
receptors containing specific subunits, e.g., a4, and through a lack of
positive modulation of
receptors containing at, e.g., traditional GABAergic mechanisms such as
benzodiazepines.
Increased action via an increase of GABA release at synapses would lead to
increased inhibition via
a pre-synaptic mechanism and thus would be expected to lead to a decrease in
anxiety and seizure
frequency. Compounds whose activity shows these features may also reduce pain,
especially
neuropathic pain. Inhibition is the only effect observed at a4, a5, and a6-
containing receptors. Both
these effects may require a y subunit, as 5-containing receptors are believed
to be unaffected by
compounds described herein.
Examples 175-179
Assessment of Therapeutic Potential of Compounds in Alleviating Anxiety
Example 175
Fear Potentiated Startle Paradigm
[0299] Purpose. To evaluate the effects of test compounds in models of anxiety
in rats, test
compounds are assessed in the fear potentiated startle paradigm (FPS) model of
anxiety.
[0300] FPS Design. FPS model is a commonly used assessment of therapeutic
value of anxiolytic
compounds in the rat. Rats may receive a 30 min period of habituation to the
FPS apparatus. 24-hr
later baseline startle amplitudes are collected. The rats will divided into
two matched groups based
on baseline startle amplitudes. Following baseline startle amplitude
collection, 20 light/shock
pairings are delivered on 2 sessions over 2 consecutive days (i.e., 10
light/shock pairings per day).
On the final day, one group of rats may receive an injection (i.v.) of a test
compound and the other
group may receive vehicle only. Immediately following injections, startle
amplitudes are assessed
during startle alone trials and startle plus fear (light followed by startle)
trials. Fear potentiated
startle (light-Fstartle amplitudes minus startle alone amplitudes) are
compared between the treatment
groups.
[0301] Animals are trained and tested in four identical stabilimeter devices
(Med-Associates).
Briefly, each rat was placed in a small Plexiglas cylinder. The floor of each
stabilimeter consists of
four 6-mm-diameter stainless steel bars spaced 18 mm apart through which shock
can be delivered.
Cylinder movements result in displacement of an accelerometer where the
resultant voltage is
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proportional to the velocity of the cage displacement. Startle amplitude is
defined as the maximum
accelerometer voltage that occurs during the first 0.25 sec after the startle
stimulus are delivered.
The analog output of the accelerometer is amplified, digitized on a scale of 0-
4096 units and stored
on a microcomputer. Each stabilirneter is enclosed in a ventilated, light-,
and sound-attenuating box.
All sound level measurements are made with a Precision Sound Level Meter. The
noise of a
ventilating fan attached to a sidewall of each wooden box produces an overall
background noise
level of 64 dB. The startle stimulus are a 50 ms burst of white noise (5 ms
rise-decay time)
generated by a white noise generator. The visual conditioned stimulus ("CS")
used is illumination of
a light bulb adjacent to the white noise source. The unconditioned stimulus is
a 0.6 mA foot shock
with duration of 0.5 sec, generated by four constant-current shockers located
outside the chamber.
The presentation and sequencing of all stimuli is under the control of the
microcomputer.
[0302] FPS procedures consists of 5 days of testing; during days 1 and 2
baseline startle responses
are collected, days 3 and 4 light/shock pairings are delivered, day 5 testing
for fear potentiated startle
was conducted.
[0303] Matching. On the first two days all rats are placed in the Plexiglas
cylinders and 3 min later
presented with 30 startle stimuli at a 30 sec interstimulus interval. An
intensity of 105 dB is used.
The mean startle amplitude across the 30 startle stimuli on the second day is
used to assign rats into
treatment groups with similar means.
[0304] Training. On the following 2 days, rats are placed in the Plexiglas
cylinders. Each day
following 3 min after entry 10 CS-shock pairings are delivered. The shock is
delivered during the
last 0.5 sec of the 3.7 sec CSs at an average intertrial interval of 4 min
(range, 3-5 min).
[0305] Testing. Rats are placed in the same startle boxes where they are
trained and after 3 min are
presented with 18 startle-eliciting stimuli (all at 105 dB). These initial
startle stimuli are used to
again habituate the rats to the acoustic startle stimuli. Thirty seconds after
the last of these stimuli,
each animal may receive 60 startle stimuli with half of the stimuli presented
alone (startle alone
trials) and the other half presented 3.2 sec after the onset of the 3.7 sec CS
(CS-startle trials). All
startle stimuli are presented at a mean 30 sec interstimulus interval,
randomly varying between 20
and 40 sec.
[0306] Measures. The treatment groups are compared based on the difference in
startle amplitude
between CS-startle and startle-alone trials (fear potentiation). In general,
the compounds described
herein may affect the startle amplitude where the greater the reduction in
fear-potentiated startle, the
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more anxiolytic the test compound. Therefore the compounds described herein
may be used in
methods of treating anxiety as shown in U.S. Patent Application Publication
Nos. 2006/0089350;
2007/0149526; and 2009/0215754.
EXAMPLE 176
Contextual Fear Conditioning Model
[0307] Purpose. To evaluate the potential of the compounds described herein
into alleviate intense
anxiety in contextual fear conditioning in rats.
[0308] Design. Contextual fear conditioning involves pairing an aversive
event, in this case
moderate foot shock, with a distinctive environment. The strength of the fear
memory is assessed
using freezing, a species-typical defensive reaction in rats, marked by
complete immobility, except
for breathing. If rats are placed into a distinctive environment and are
immediately shocked, they do
not learn to fear the context. However, if they are allowed to explore the
distinctive environment
sometime before the immediate shock, they show intense anxiety and fear when
placed back into the
same environment. 13y procedurally dividing contextual fear conditioning into
two phases, one can
separately study effects of treatments on memory for the context (specifically
a hippocampus based
process) from learning the association between context and shock or
experiencing the aversiveness
of the shock (which depend upon emotional response circuitry including
amygdala). Post-Traumatic
Distress Syndrome (PTSD) in humans has been shown to be related to emotional
response circuitry
in the amygdala; for this reason contextual memory conditioning is a widely
accepted model for
PTSD.
[0309] At typical experiment may use 24 rats. Each rat may receive a single 5-
min episode of
exploration in a small, novel environment. 72-hr later they will placed into
the same environment,
and immediately they will receive a single, moderate foot-shock. 24-hours
later, 12 of the rats may
receive an injection (i.v.) of a test compound as described herein. The
remaining 12 rats may receive
an injection of the vehicle. Each rat may again be placed into the same
environment for 8-min,
during which time freezing will measured, as an index of Pavlovian conditioned
fear.
[0310] Methods. In a typical experiment, 4 identical chambers (20 X 20 X 15
cm) are used. All
aspects of the timing and control of events are under microcomputer control.
Measurement of
freezing is accomplished through an overhead video camera connected to the
microcomputer and is
automatically scored using a specialty piece of software, FreezeFrame. In
Phase 1, rats are placed
individually into the chambers for 5 minutes. Phase 2 begins 72 hours later,
when again rats are
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placed individually into the same chambers but they receive an immediate foot
shock (e.g., 1 mA for
2 s). Thirty seconds later they are removed from the chambers. Phase 3, 24
hours later, the rats are
returned to the chambers for 8 min, during which time freezing, the index of
conditioning fear is
scored. Total freezing time will analyzed in a one-way ANOVA with drug dose as
the within-
groups factor.
[0311] Compounds described herein which show significant anxiolytic effect in
this model system
may be used in a method for treating anxiety or post-traumatic stress
disorder. Exemplary use of this
model system are shown in U.S. Patent Application Publication Nos.
2006/0089350 and
2009/0215754.
EXAMPLE 177
Elevated Plus Maze
[0312] Design. The elevated plus maze (EPM) is commonly used to assess anxiety
levels in
rodents. The EPM takes advantage of the fact that when a normal rat is feeling
anxious in a novel
environment it may seek out and hide in enclosed spaces. A normal rat may
venture out into open
spaces within the new environment only when it feels less anxious. Drugs like
diazepam and
buspirone show anxiolytic effects in this task, and hence rats treated with
such drugs spend more
time within the open areas of the maze.
[0313] This experiment may employ two groups of rats. A first group of the
rats may receive an
injection (i.v) of test compound and a second group may receive an injection
of vehicle. Fµach rat
may immediately be placed on the elevated plus maze. Time spent in the open
arms of the maze will
recorded and compared between treatment groups. If the test compound reduces
anxiety in rat then
the group that received the test compound may spend more time in the open arms
than the rats that
received vehicle.
[0314] The elevated plus maze may consist of two opposing open arms, 50X10 cm,
crossed with
two opposing enclosed arms of the same dimensions but with walls 40 cm high.
Each of the four
arms is connected to one side of a central square (10X10 cm) giving the
apparatus a plus-sign
appearance. The maze will elevated 50 cm above the floor in a normally
illuminated room. The rats
are placed individually on the central square of the plus maze facing an
enclosed arm. The entire 3-
min session is videotaped and later scored. The time spent and the number of
entries into the open
and closed arms, and the number of trips made to at least the midpoint down
the open arms is
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recorded. An arm entry is defined as placement of all four paws onto the
surface of the arm. This
model is described in U.S. Patent Application Publication No. 2006/0025387.
EXAMPLE 178
Marble Burying Test in the Mouse
[0315] This method detects anxiolytic/tranquillizing activity and was
described by Broekkamp, et al.
(1986) Eur. J. Pharmacol. 126: 223-229. Mice exposed to novel object (marbles)
will bury them in
the sawdust floor covering. Anxiolytics decrease the number of marbles buried
at non-sedative
doses.
[0316] Mice will individually placed in transparent plastic cages (33 x 21 x
18 cm) with 5 cm of
sawdust on the floor 25 marbles grouped in the centre of the cage. The cage
will covered with an
inverted plastic cage. Each test cage, together with the marbles, will
impregnated with mouse odor
before-hand by leaving 10 mice in the cage for 15 minutes. These mice then
play no further role in
the experiment.
[0317] The number of marbles covered by sawdust (2/3 or more) will counted at
the end of a 30
minute test. 12 mice will studied per group. The test will performed blind.
Each of two (2) test
substances will evaluated at 3 doses, administered i.p. 30 minutes before the
test, and compared with
a vehicle control group. Clobazam (8 mg/kg i.p.) under the same experimental
conditions will used
as reference substance. The experiment may include 8 groups.
TABLE 6: Anxiety Model Results for Seletected Compounds
Compound .= Dose : . Number of Marbles covered by
'Number of marbles covered
(mg/kg) sawdust sawdust as compared
to
.................................(mean . . control
5011 50 6.3 ( 2.4)
-71%
6001 50 12.8 ( 2.7)
-41%
6008 50 8.8 ( 3.2)
-59%
CLOBAZAM 8 2.8 ( 2.0)
-87%
TABLE 7: Anxiety Model Results for Seletected Compounds
Compound Dose Number, of Marbles covered by
Number of marbles covered by::'=
(mg/kg) sawdust sawdust as compared
to
. Imean s.e.m.) .1
control
2014 10.5 26.3 ( 0.3)
-10%
2014 31.5 25.9 ( 0.3)
-51%
2014 105.1 27.3 ( 0.4)
-98%
4011 9.4 27.1 ( 0.5)
-14%
4011 28.2 25.8 ( 0.4)
-1%
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4011 94.1 26.8 ( 0.4) -17%
CLOBAZAM 8 26.2 ( 0.3) -77%
[0318] Compounds described herein may decrease the number of marbles buried at
non-sedative
doses and thus may be useful as anxiolytics.
EXAMPLE 179
Light Dark Box Test
[0319] This method detects anxiolytic activity and was described by Crawley
(1981) Pharmacol.
Biochem. Behav. 15: 695-699. Anxiolytics increase the time spent in the light
compartment and
decrease the number of crossings.
[0320] Animals will placed into the light compartment of a 2-compartment box
with one half light
and open (25 x 27 x 27 cm) and the other half dark and closed (20 x 27 x 27
cm). The time spent in
each compartment as well as the number of times the animal crosses from one
side to the other will
scored during a 3-minute test. 10 mice will studied per group. The test will
performed partially blind
(apart from positive control).
[0321] The compounds will evaluated at 3 doses, administered i.p. 30 minutes
before the test, and
compared vehicle control group. Clobazam (16 mg/kg i.p.) will administered 30
minutes before the
test as a reference substance. The experiment may include 8 groups.
TABLE 8: Results for Light/Dark Box Test
Compound Dose Time Spent in Light r Time Spent in Number of
(mg/kg) Compartment as compared to Crossings as compared to
, control control
2014 10.5 -20% +8%
2014 31.5 +85% -37%
2014 105.1 +87% -84%
5009 11 0% +16%
5009 32.9 +54% -33%
5009 109.6 +98% -72%
CLOBAZAM 16 +104% -42%
[0322] Two selected compounds described herein (e.g., 2014 and 5009) increase
the time spent in
the light compartment and decrease the number of crossings. Therefore, the
compounds described
herein may be usedful as anxiolytics in the treatment of anxiety disorders.
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Examples 180-183
Analgesic/Anti-inflammatory Activity Models
Experimental Models of Pain
[0323] Experimental models of pain include tests of response thresholds to
high intensity stimuli
(acute pain tests) and changes in spontaneous or evoked behavioral responses
in animals with
peripheral injury or inflammation (persistent pain models). Acute thermal pain
is modeled by the
hot-plate and tail-flick test, while persistent pain can be modeled by the
formalin test. See Bannon
and Malmberg "Models of Nocipetion: Hot-Plate, Tail-Hick, and Formalin Tests
in Rodents." Curr.
Protoc. Neurosci. 41:8.9.1-8.9.16 for protocols for all three of these tests,
including preparation of
animals (rats or mice), administration of a compound being tested for its
analgesic properties and
data collection.
EXAMPLE 180
Formalin Paw Test (late phase)
[0324] The method described herein detects analgesic/anti-inflammatory
activity, generally used to
test compounds for pain relief, in particular diabetic neuropathy or
nociceptive neuropathy. See
Wheeler-Aceto, et al. (1991) Psychopharmacology 104: 35-44.
[0325] Mice are given an intraplantar injection of 5% formalin (251.11) into
the posterior left paw.
This treatment induces paw licking in control animals. Mice are briefly
observed at 1 minute
intervals between 15 and 50 minutes after the injection of formalin and the
number of occasions that
the mice are observed licking the injected paw is recorded. There are 10 mice
per group and the test
is performed "blind."
[0326] Compounds as described herein will evaluated at 3 doses each,
administered i.p. 30 minutes
before the test (i.e., 15 minutes before formalin), and compared with a common
vehicle control
group. Morphine (8 mg/kg i.p.) or gabapentin (100 mg/kg i.p.) or venlafaxine
(32 mg/kg i.p.),
administered under the same experimental conditions, is used as reference
substance. Data is
analyzed by comparing treated groups with vehicle control using unpaired Mann-
Whitney U tests.
[0327] The data recorded for each animal is the amount of time(s) spent
licking the affected hind
paw in a two minute period. These two minute periods occur at five minute
intervals and continue
for 45 minutes. Plotting the time spent licking versus time reveals the
characteristic biphasic
response. From this plot, the area under the curve (AUC) for each animal
during both the acute and
inflammatory stages are calculated. The AUC for each phase is shown for both
control and drug-
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treated animals. The AUC for each drug-treated animal is compared to the
average result from the
control group, yielding an average percent of control. Significant reductions
in this number indicate
a reduction in licking and a reduction of perceived pain.
[0328] Several compounds described were tested in a Formalin Paw Test at a
does of 250 jimol/kg
using vehicle 1 control (0.2% HPMC in physiological saline), Gabapentin (100
mg/kg i.p.) as a
reference substance, and vehicle 2 (5% dimethylacetamide/50% PEG 400/45% H20).
TABLE 10: Analgesic effects of selected compounds
, Compound
(15-50 minutes after formalin)Licking Score
NTP-4011
¨55%
NTP-3034
¨61%
NTP-6008
¨99%
NTP-4005
¨66%
NTP-2014
¨98%
NTP-6002
¨82%
Gabapentin
¨76%
Vehicle 2
¨90%
[0329] Several compounds described herein show a reduction in perceived pain
and thus may be
used in methods to treat and/or prevent (prophylactic) for pain.
EXAMPLE 181
Taxol Induced Neuropathy Model
[0330] Peripheral neuropathies are chronic conditions that arise when nerves
are damaged by
trauma, disease, metabolic insufficiency, or by certain drugs and toxins. The
sensory disturbances
associated with chemotherapeutic agents, such as paclitaxel (Taxo10), range
from mild tingling to
spontaneous burning, typically in the periphery such as the hands and feet.
Symptoms become more
intense with continued therapy and can lead to weakness, ataxia, numbness and
pain, limiting the
dose and/or treatment with the chemotherapeutic agent.
[0331] Gabapentin, 100 mg/kg, IF is able to mitigate the mechanical allodynia
seen as a result of the
Taxol-induced neuropathic pain. Similarly, rats treated with compounds
described herein are
believed to show a significant improvement in allodynia when compared to the
vehicle control
group.
[0332] Test articles are administered intraperitoneally in dose volumes of 10
mL/kg body weight.
Preparations are made freshly for each day of administration.
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[0333] The reference article, Gabapentin, is formulated in saline to a
concentration of 100 mg/mL
and delivered subcutaneously at a dose volume of 1 mL/kg body weight (for a
dosing concentration
of 100 mg/kg). Male Sprague Dawley rats will used.
[0334] Allocation to Treatment Groups. For inclusion into the study (first
portion), the animals
have a baseline thermal paw test, which is measured prior to Taxol injections.
Animals with a
thermal paw score greater than 15 seconds are excluded from study.
[0335] All animals that may receive Taxol are tested for thermal
hyperalgesia. Animals need to
have at least a 20% drop from baseline for inclusion into the treatment
segment of the study. All
animals underwent a baseline pre-dose von Frey test, which is measured prior
to Taxol injection.
For inclusion into the study, the animals needed to have a baseline von Frey
score above 12.
[0336] All animals administered Taxol are tested for mechanical allodynia
using von Frey.
Animals receiving a score of 13 or below are allocated to treatment groups.
The mechanical
allodynia scores for each group are reviewed to ensure the mean values and
standard deviations are
homogeneous. Rats are allocated to treatment groups.
[0337] All animals are administered Taxol , 2 mg/kg, IP at a dose volume of 1
mL/kg, on Days 1,
3, 5 and 7.
[0338] All animals may receive a single intraperitoneal injection of test
compound. All animals may
receive an IP injection of vehicle or test compound 30 min prior to mechanical
allodynia testing.
Animals are dosed at a volume of 10 mL/kg.
[0339] Control animals may receive an IF injection of Gabapentin 90 minutes
prior to mechanical
allodynia testing. Animals are dosed at a volume of 1 mL/kg.
[0340] Behavioral Testing¨Acclimation. Twice prior to baseline testing, the
animals underwent
acclimation to the mechanical allodynia apparatus. This habituated the rats to
the testing devices so
they are calm at the time of testing.
[0341] Mechanical Allodynia (von Frey). All animals undergo von Frey testing
for mechanical
allodynia. On testing days, the animals are returned to the chambers and
allowed approximately 15
minutes to explore their surroundings prior to testing. A filament is applied
to the left hind paw.
[0342] The group mean results are analyzed versus the vehicle control group
using a two way
ANOVA, followed by a Bonferroni post-hoc test. Individual groups are tested
pre and post dose
using a paired t-test.
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[0343] Therefore, compounds described herein act parasynaptically and will
administered to treat
neuropathic pain without the unwanted side effects usually associated with
GABAergic compounds
(e.g., sedation from benzodiazepines). Further, treatment with compounds
described herein are
expected to have similar effects.
EXAMPLE 182
Tail Flick Test Model of Nocipetion
[0344] Overview. The mouse tail-flick assay is a well-accepted model of acute
thermal pain in
which a number of clinically relevant opioid analgesics produce moderate to
full efficacy on several
different pain-related measures. The mouse model also requires smaller amounts
of drug and is a
more rapid initial screen for antinociceptive activity compared to the chronic
pain models. This
model of pain is also described in U.S. Patent Application Publication Nos.
2006/0089550 and
2006/0025387.
[0345] Tests for Antinociception. Efficacy of the test compound may be
assessed using the 52 C
warm water tail-flick test. The latency to the first sign of a rapid tail-
flick is taken as the behavioral
endpoint (Jannsen et al., 1963). Each mouse is first tested for baseline
latency by immersing its tail
in the water and recording the time to response. Mice not responding within 5
seconds are excluded
from further testing. Mice are then administered the test compound and tested
for antinociception at
various time points afterwards. Antinociception is calculated by the following
formula: %
Antinociception = 100 x (test latency-control latency)/(10-control latency). A
maximum score is
assigned (100%) to animals not responding within 10 seconds to avoid tissue
damage.
[0346] Subjects: Male CD-1 (25-35 g, Charles River) mice may be used for all
studies. Mice will
housed in groups of five in Plexiglas chambers with food and water available
ad libitum. All
animals are maintained on a 12 hr light/dark cycle (lights on at 7:00 AM) in a
temperature- and
humidity-controlled animal colony. Only animals in good health are
acclimatized to laboratory
conditions and are used in the study. The acclimation period to the vivarium
will a minimum of 7
days. All animal experiments may be performed under an approved protocol in
accordance with
institutional guidelines and in accordance with the Guide for the Care and Use
of Laboratory
Animals as adopted and promulgated by the National Institutes of Health.
[0347] Drugs and Injections. Morphine sulfate (Mallinckrodt, St. Louis, MO)
may be dissolved in
physiological saline. Injections will made using a 1-mL syringe with a 30-
gauge needle at a volume
of 10 ml/kg body weight. Animal body weights will measured on the morning of
testing. Mice are
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firmly grasped by the nape of the neck and the tail is tucked between the last
two fingers and palm of
the technician's hand. The back of the mouse is arched slightly backwards
exposing the abdominal
region. The needle is inserted through the skin and abdominal musculature into
the peritoneal cavity
just off of the midline. Mice are immediately returned to the holding cage
until behavioral testing.
[0348] Total Number of Animals: Each experimental group may consist of 8 mice
with a total of
approximately 260 mice needed to complete the proposed studies (30 compounds x
1
dose/compound x 8 mice/group plus controls).
[0349] Housing: Automatically controlled environmental conditions are set to
maintain temperature
at 20-24 C with a relative humidity (RH) of 30-70%, a 12:12 hour light:dark
cycle and 15-30 air
changes/h in the study room. Temperature and RH are monitored daily. During
acclimation and
throughout the entire study duration, animals are housed within a limited
access rodent facility and
kept in groups of 5 mice/polypropylene cages (23 x 17 x 14 cm), fitted with
solid bottoms and filled
with wood shavings as bedding material. Individual cages are suspended in a
stainless steel rack
system with each cage having its own water sipping tube and food hopper. Mice
may have ad
libitum access to food (Harlan Teklad Global 2018) and water except during the
formal drug
administration and testing procedures.
[0350] Overall Experimental Design: On the morning of Day 1, mice are marked
and weighed,
and then baselined for thermal latencies in the 52 C tail-flick assay. Test
compounds are then
injected and the mice are rested for thermal latencies at 10, 20, 30, 45, 60,
90, 120 and 180 minutes
post-injection (if a drug effect drops below 20% for the group average then
the testing is halted for
that group). All compounds may be dissolved in 0.2%
hydroxypropylmethylcellulose (HPMC) in
physiological saline to yield final concentrations of between 5-20 mg/ml (pH
¨7.4). Tween 80 (1-2
drops) may be added to aid in soulbilization of compounds. A similar vehicle
will used as a control.
Morphine sulphate may be dissolved in physiological saline.
[0351] Evidence of the ability of compounds described herein to alleviate pain
will seen as a higher
pain threshold than the group that received vehicle as tested. The results of
selected compounds
tested in the tail flick pain model (e.g., 4012, 4018, 5009, 5011, 6002, 6008,
6009, 7009, 10001) are
presented in FIGURE 9. The showing of increasing the pain threshold of the
treated mice suggests
that the compounds described herein may be useful as analegiscs in the
treatment of pain.
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EXAMPLE 183
Chung model of neuropathy
[0352] Design. Spinal nerve ligation is performed under isoflourane anesthesia
with animals placed
in the prone position to access the left L4¨L6 spinal nerves. Under
magnification, approximately
one-third of the transverse process is removed. The L5 spinal nerve is
identified and carefully
dissected free from the adjacent L4 spinal nerve and then tightly ligated
using a 6-0 silk suture. The
wound is treated with an antiseptic solution, the muscle layer is sutured, and
the incision is closed
with wound clips. Behavioral testing of mechanical paw withdrawal threshold
takes place within a
3-7 day period following the incision. Briefly, animals are placed within a
Plexiglas chamber
(20X10.5X40.5 cm) and allowed to habituate for 15 min. The chamber is
positioned on top of a
mesh screen so that mechanical stimuli can be administered to the plantar
surface of both hindpaws.
Mechanical threshold measurements for each hindpaw are obtained using an
up/down method with
eight von Frey monofilaments (5, 7, 13, 26,43, 64, 106, and 202 mN). Each
trial begins with a von
Frey force of 13 mN delivered to the right hindpaw for approximately 1 sec,
and then the left
hindpaw. If there is no withdrawal response, the next higher force is
delivered. If there is a
response, the next lower force is delivered. This procedure is performed until
no response is made at
the highest force (202 mN) or until four stimuli are administered following
the initial response. The
50% paw withdrawal threshold for each paw is calculated using the following
formula:
[Xth]log=[vFr]log.-i-ky where [vEr] is the force of the last von Frey used,
k=0.2268 which is the
average interval (in log units) between the von Frey monofilaments, and y is a
value that depends
upon the pattern of withdrawal responses. If an animal does not respond to the
highest von Frey hair
(202 mN), then y=1.00 and the 50% mechanical paw withdrawal response for that
paw is calculated
to be 340.5 mN. Mechanical paw withdrawal threshold testing is performed three
times and the 50%
withdrawal values are averaged over the three trials to determine the mean
mechanical paw
withdrawal threshold for the right and left paw for each animal. This model is
described in U.S.
Patent Application Publication No. 2006/0025387.
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EXAMPLES 184-186
SEIZURE MODELS
EXAMPLE 184
Mouse model of Mesial Temporal Lobe Epilepsy (MTLE)
[0353] Purpose. To test the anti-seizures properties of compounds described
herein, compounds
may be tested for their ability to suppress seizures in a kainate model of
epilepsy.
[0354] Methods. Using stereotaxic techniques under general anaesthesia,
intrahippocampal
injection of kainate (1 nmole in 50 n1) will performed on adult mice (e.g.,
C57/B16). Mice may then
be implanted with a bipolar electrode in the dorsal hippocampus and 3 cortical
rnonopolar
electrodes. After 4 weeks following kainate injection, the mice will injected
with test compounds
(e.g., two injections per week). Drug conditions will counter-balanced, the
animals being used as
their own controls. Digital EEG recordings will performed in freely moving
animals for 20 minutes
pre-injection and 20 minutes between 20 and 40 minutes post-injection. The
effects of the injected
compound will compared versus reference period. Several compounds described
herein were tested
in an MTLE model at a dose of at least about 20-75 mg/kg.
TABLE 9: MTLE Model Results for Selected Compounds
'Compound Dose Reduction in .. Reduction in
:.:Slight increase in
(mg/kg) . number of number of number of
hippocampal hippocampal hippocampal
discharges by more diseharge.s by less discharges
than 50% : than 50%
NTP-4007 23
X
NTP-5009 27 X
NTP-4011 47 X
NTP-4008 52 X
NTP-6008 59 X
NTP-6002 61 X
NTP-3034 52 X
NTP-4005 59 X
NTP-5013 57 X
NTP-7013 67 X
NTP-3003 64 X
NTP-3004 64 X
NTP-4012 61 X
NTP-3032 61 X
NTP-4018 70 X
NTP-4020 66 X
NTP-5009 55 X
NTP-5011 54 X
NTP-6001 56 X
NTP-6006 56 X
NTP-6009 59 X
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NTP-7009 65 X
NTP-7049 52 X
NTP-10001 51 X
[0355] Several compounds described herein decrease the cumulative duration and
number of
hippocampal discharges in this model of mesial temporal lobe epilepsy.
Therefore, the compounds
described herein may be used in therapeutic methods to treat seizures.
Example 185
Epileptiform Discharges in Hippocampal Slices
[0356] During these studies, spontaneous epileptiform activity may be elicited
by a variety of
treatments. Sprague-Dawley rats (males and females; 25-35 days old) will
decapitated, the top of
the skull is rapidly removed, and the brain will chilled with ice-cold
oxygenated slicing medium. A
slicing medium used will a sucrose-based artificial cerebrospinal fluid
(sACSF) consisting of 220
mM sucrose, 3 mM KC1, 1.25 mM NaH2PO4, 2 mM MgSO4, 26 mM NaHCO3, 2 mM CaC12,
and 10
mM dextrose (295-305 mOsm). A hemisphere of brain containing hippocampus will
blocked and
glued (cyanoacrylic adhesive) to the stage of a Vibroslicer (Frederick Haer,
Brunsick, ME).
Horizontal or transverse slices 400 gm thick are cut in 4 C, oxygenated (95%
02; 5% CO2) slicing
medium. The slices are then immediately transferred to a holding chamber where
they remained
submerged in oxygenated bathing medium (ACSF) consisting of 124 mM NaC1, 3 mM
KC1, 1.25
mM NaH2PO4, 2 mM MgSO4, 26 mM NaHCO3, 2 mM CaC12, and 10 mM dextrose (29-305
mOsm). The slices can be held at room temperature for at least 45 minutes
before being transferred
to a submersion-style recording chamber. In the recording chamber, the slices
will perfused with
oxygenated recording medium at 34-35 C. All animal procedures should be
conducted in
accordance with NIH animal care guidelines. In most slice experiments,
simultaneous extracellular
field electrode recordings are obtained from CA1 and CA3 as described in U.S.
Patent Nos.
6,495,601 and 7,214,711.
[0357] Hippocampal slices treated with the test compounds may show less
epileptiform activity,
indicative of anti-seizure properties. Compounds described herein may be used
in methods to treat
and/or prevent (prophylactic) for seizures, seizure disorders, epilepsy,
epileptic seizures, and other
neurodegenerative disorders (e.g., those neurodegenerative disorders which
involve seizures).
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Example 186
In vitro Hippocampal Recordings of miniature and spontaneous
inhibitory post-synaptic currents (mIPSCs and sIPSCs)
[0358] Ion flux in neuronal cells are measured using standard techniques.
Kandel and Schwartz
Principles of Neural Science 2"I Edition (1985), see, e.g., pages 128-131.
Recording is performed in
vitro in hippocampal slices (CAI pyramidal cell layer). For recording GABAA-
IPSCs, glutamatergic
and GABAB transmission is blocked by adding DNOX (50 RM), AP-5 (50 M), and
SCH50911 (20
mM) into the medium. The intracellular solution comprised CsC1 and OX314.
[0359] Compounds may be tested for their ability to increase GABAA inhibitory
drive, such as a
marked increase in spontaneous IPSCs or in miniature IPSCs in a hippocampal
slice model where
the compound consistently shows a significant decrease in the time between
inhibitory events (e.g.,
increased frequency of events).
[0360] Data demonstrating that the interval between miniature and/or
spontaneous inhibitory post-
synaptic currents (mIPSCs and sIPSCs, respectively) events are substantially
decreased in the
presence of the compound indicate a highly significant increase in the
frequency of inhibitory
events. Such data suggest a pre-synaptic mechanism increasing the release of
GABA from the
neurons by the action of compounds described herein.
[0361] Hippocampal Slice Preparation¨ Young adult male Sprague-Dawley rats
weighing 220-
250 g at time of use were housed in groups of 4 in an air conditioned room on
a 12 hour light/dark
cycle with food and water available ad libituin. On the day of experiments,
animals were terminally
anaesthetised using isofluorane, cervically dislocated, and decapitated. The
brain was removed and
300-400 p.m thick hippocampal slices cut using a Leica VT1000S.
[0362] Electrophysiological recording¨ Slices were maintained in artificial
cerebrospinal fluid
(aCSF) at room temperature for 1 hour after slicing before commencing
electrophysiological
recordings. After this period, individual slices were transferred to a custom-
built chamber
continuously perfused with aCSF at a rate of 4-10 ml/min of the following
composition (mM):
NaC1, 127; KC1, 1.9; KH2PO4, 1.2; CaC12, 2.4; MgCl2, 1.3; NaHCO3, 26; D-
glucose, 10; equilibrated
with 95% 02-5% CO2. Whole-cell patch-clamp recordings were performed at room
temperature
(17-21 C) from hippocampal pyramidal CAI neurons using an Axoclamp 700B
amplifier using the
'visualised' version of the patch clamp technique. Patch pipettes had
resistances of between 3 and 8
MO when filled with intracellular solution of the following composition (mM):
caesium chloride,
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150; EGTA-Na; HEPES, 10; Na2ATP. All experiments were carried out in the
presence of
Tetrodotoxin (TTX, 1 1.1M), NBQX (10 p,M), D-AP-5 (10 RM) and CGP55845 (GABAB
antagonist,
200 nM) to eliminate action potentialdependent synaptic activity and isolate
GABAA receptor-
dependent synaptic responses.
[0363] Test Compounds¨ 3034, 6008, 6009, and 7049 were prepared as 5 mM stock
in DMSO and
serially diluted to the required concentrations in aCSF immediately prior to
use. TTX (1 mM),
Bicuculline (10 mM), NBQX (10 mM), CGP55845 (10 mM) and AP5 (10 mM) were all
obtained
from Ascent Scientific and prepared as stock solutions in DMSO or ddH20 as
appropriate. All
compounds were stored at ¨20 C prior to use. Test compounds were cumulatively
applied for a
minimum of 10 minutes.
[0364] Analysis¨ All analysis was conducted using Excel (Microsoft), Clampfit
(MDS
Technologies) and Mini-analysis (Lavasoft) software. mIPSC analysis was
performed on 60s traces
using a threshold of ¨10 pA and an area of ¨150 fC/s for synaptic currents.
[0365] Three select compounds (e.g., 3034, 6009, 7049) were tested in synaptic
transmission in
adult rat hippocampal neurons in vitro. See FIGURES 10-12. Compounds 3034,
6009 increased
mIPSC frequency in a concentration-dependent manner. Compounds 3034, 6008,
6009, and 7049
decreased mIPSC inter-event interval in a concentration manner. Compounds
3034, 6008, 6009, and
7049 had no effect on mIPSC amplitude. These data indicate an increase in
presynaptic inhibition
corresponding to the increased mIPSC frequency. The compounds did not increase
the mIPSC
amplitude indicative that the compounds do not affect post-synaptic
inhibition. Further, this data is
indicative of these compounds increasing GABA inhibitory drive.
[0366] Compounds described herein which act parasynaptically may be
administered at high doses
(e.g., 100 mg/kg) without the unwanted side effects usually associated with
GABAergic compounds
(e.g., sedation from benzodiazepines). Compounds may show anticonvulsant
activity and may be
useful as therapeutics for treating seizure disorders without these unwanted
side effects.
EXAMPLE 187
6 Hz Psychomotor Test in the Mouse
[0367] The 6Hz psychomoter test method detects anticonvulsant activity and was
described by
Brown, et al. (1953) J. Pharmacol. Exp. Ther. 107: 273-283. Mice will
administered a rectangular
current (e.g., 32 mA, rectangular pulse: 0.2 ms pulse width, 3 s duration, 6
Hz) via corneal electrodes
connected to a constant current shock generator. The results for the number of
seizures as reflected
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by forelimb clonus and Straub-tail are recorded during 30 seconds following
current administration.
Forelimb clonus is scored as absent (0), mild (1) and strong (2) whereas
Straub tail is rated as absent
(0) or present (1).
[0368] Test substances will evaluated at 1 dose, administered i.p. 30 minutes
before the test and
compared with a vehicle control group. 15 mice will studied per group. The
test will performed
blind. Diazepam (2 mg/kg i.p.) will administered under the same experimental
conditions and used
as reference substance. Compounds which lower the bumber of seizures
experienced by the mouse
are indicative of compounds with anticonvuslant activity that may be useful in
the treatment of
seizures.
EXAMPLE 188
Amphetamine Hyperactivity Test in the mouse
[0369] This method detects antipsychotic and antiparkinson activity and was
described by Costa11, et
al. (1977) Brain Res. 123: 89-111, and uses an activity meter similar to that
described by Boissier
and Simon (1965) Arch. Int. Pharmacodyn. 158: 212-221.
[0370] Amphetamine induces hyperactivity in this test situation. Hyperactivity
is antagonized by
classical and atypical antipsychotics acting on dopaminergic systems at the
limbic level, and is
potentiated by antiparldnson drugs. Mice were injected with d-2014mphetamine
(3 mg/kg i.p.) and
are immediately placed in the activity meter. The activity meter may consist
of 24 covered Plexiglas
cages (20.5 x 10.5 x 18 cm) contained within a darkened cabinet and connected
to silent electronic
counters. Each cage was equipped with four photocell assemblies (two at each
end of the cage) 2.5
cm above the floor, in order to measure the number of movements by each animal
(one per cage) in
the horizontal plane. Seven additional photocell assemblies will placed at
even intervals 9.5 cm
above the floor along the long wall to record rearing. The number of
(horizontal) crossings by each
animal (one per cage) from one pair of photocells to the other will recorded
by computer in 10-
minute intervals for 30 minutes. A similar procedure will utilized for
recording of rearing, except
that individual photobeam breaks are recorded. The scores for activity and
rearing will recorded by
computer over 10-minute intervals and cumulated over a 30-minute period. 10
mice were studied
per group. The test was performed blind.
[0371] Each compound was evaluated at a dose of 50 mg/Kg, administered i.p. 30
minutes before
amphetamine, and compared with a vehicle control group. The experiment may
also include a
control group not treated with amphetamine. Haloperidol (0.25 mg/kg i.p.) may
be administered
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under the same experimental conditions as a reference substance. The
experiment may include 9
groups.
TABLE 11: Hyperactivity Test Results
Compound Number of Crossings Compared with Number of rears Compared
with
(0-30') Amphetamine (0-30') Amphetamine
fmean man.] ' Control [mean s.ean.] Control
Amphetamine Control 313.7 ( 54.4) +572% 80.7 ( 21.6)
¨33%
2014 232.2 ( 38.8) ¨26% 44.0 ( 15.7) ¨45%
5009 159.2 ( 33.4) ¨49% 32.7 ( 8.6) ¨59%
6008 137.8 ( 25.2) ¨56% 33.8 ( 14.4) ¨58%
HALOPERIDOL 110.3 ( 21.0) ¨65% 111.3 ( 34.3)
+38%
[0372] The selected compounds described herein were tested in the amphetamine-
induced
hyperactivity test and show a decrease in the number of crossings and number
of rears. Therefore
the compounds described herein show antipsychotic and antiparkinson activity
and may be useful in
the treatment of anxiety disorders, ADHD, and Parkinson's disease as well as
other movement
disorders that involve bradykinesia such as Huntington's Disease.
Example 189
Animal Model of Amphetamine Sensitization (Addiction)
[0373] Purpose. The therapeutic usefulness of compounds in the treatment of
behavior disorders
may be examined by measuring the ability of a compound to reverse the symptoms
of amphetamine
sensitization in rats.
[0374] Method. Amphetamine sensitization will induced in animals. Following
sensitization, the
rats are divided into two equal groups. One group receives treatment with
compounds and the other
half receives vehicle. All rats may then be given a challenge injection of
amphetamine. Open field
motor activity will monitored. If a compound reduces or blocks amphetamine
sensitization, the
group that received compounds prior to the amphetamine challenge exhibits
shorter distances and
fewer total rears.
[0375] Following three days of handling, the animals receive daily
intraperitoneal (i.p.) injections of
1.5 mg/kg amphetamine hydrochloride (injection volume 1.0 ml/kg) for 5 days
(amphetamine-
amphetamine group). Amphetamine may be freshly diluted with saline (0.9%)
every morning
(injections performed between 10:00 and 12:00 h). The fifth day of treatment
with amphetamine
will followed by withdrawal for 48 h. Following the 48 hours withdrawal,
eight, of the rats receive
an injection of compounds (i.v) and eight receive an injection of vehicle
(i.v). The rats then receive
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a challenge injection of amphetamine (1.5 mg/kg) and are monitored for
locomotor activity in an
open field. All injections except the challenge injection are administered in
the rats' home cage.
[0376] Locomotor activity will measured in an open field for 120 min following
the amphetamine
challenge. Total distance traveled and number of rears are automatically
recorded and compared
between groups using one-way analysis of variance. This model is described in
U.S. Patent
Application Publication No. 2006/0025387.
[0377] Although the invention has been described in some detail by way of
illustration and example
for purposes of clarity of understanding, it will be obvious that certain
changes and modifications
will practiced within the scope of the appended claims. Modifications of the
above-described modes
for carrying out the invention that are obvious to persons of skill in
medicine, pharmacology,
microbiology, and/or related fields are intended to be within the scope of the
following claims.
[0378] All publications (e.g., Non-Patent Literature), patent application
publications, and patent
applications mentioned in this specification are indicative of the level of
skill of those skilled in the
art to which this invention pertains. All such publications (e.g., Non-Patent
Literature), patent
application publications, and patent applications are herein incorporated by
reference to the same
extent as if each individual publication, patent, patent application
publication, or patent application is
specifically and individually indicated to be incorporated by reference.
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