1. health and fitness
2. disease
3. chronic pain

; 13 MAY 2006
t 3 3 11 KOL -NP 12006
.\t: 2005/042497
PCTlUS2004/036297
t
,
TECHNICAL FIEID OF THE INVENTION
[0001]
sodium
The present invention
channels.
comprising
The invention
the compounds
relates to compounds
also provides
of the invention
useful as inhibitors
pharmaceutically
and methods
of voltage-gated
acceptable
compositions
of using the compositions
in the
treatment of various disorders,
BACKGROUND
OF THE INVENTION
[0002] Na channels are central to the generation of action:'potentials
neurons ~d myocytes.
,T~~y play key. roles
in excitable
in all excitable cells such as'
tis~u~ including brain ..smooth muscJes .
of the' gastrdi:ritestin~
tract, skeletal nhiscle,. the ,penp~e.rar nerVouB system,' spinaJ. cord' and,
.
."
.
.
'
airway. As such they play key roles in a variety of disease states such as epilepsy (See, Moulard,
'.
.'
B. and D. Bertrand (2002) "Epilepsy and sodium channel blockers" Expert Ooin. Ther. Patents
12(1): 85-91», pain (See, Waxman, S. G., S. Dib-Hajj, et ai, (1999) "Sodium channels and pain"
Proc Natl Acad Sci USA
"Voltage-gated
96(14): 7635-9 and Waxman, S. G., T. R. Cummins, et aI. (2000)
soqium channels and the ,molecular pathogenesis
of pain: a review" J Rehabil
Res Dev 37(5): 517-28), myotonia (See, Meola, G. and V. Sansone (2000) ''Therapy in myotonic
disorders and in muscle channelopathies"
Neurol Sci 21(5): S953-61 and Mankodi, A. and C. A.
Thornton (2002) "Myotonic syndromes" CUI! Opin NeuroI15(5):
M. H., J. A. Kearney, et al. (2002) "Mutations
of voltage-gated
545-52), ataxia (See, Meisler,
sodium channels in movement
disorders and epilepsy" Novartis Found SYillP 241: 72-81), multiple sclerosis (See, Black, J. A.,
- 11--
~
PCT/uS2004/036297
2005/042497
sodium channel SNS is abnormally expressed
S. Dib-Hajj, et al. (2000) "Sensory neuron-specific
in the brains of mice with experimental
sclerosis" Proc Natl Acad Sci USA
allergic encephalomyelitis
and humans with multiple
97(21): 11598-602, and Renganathan,
M., M. Gelderblom,
et aI. (2003) ''Expression of Na(v)1.8 sodium channels perturbs the firing patterns of cerebellar
purkinje
cells" Brain Res 959(2): 235-42), irritable bowel (See, Su, X., R. E. Wachtel, et al.
(1999) "Capsaicin sensitivity and voltage-gated
rat dorsal root ganglia" Am J Physiol277(6
sodium currents in colon sensory neurons from
and Laird, J. M., V. Souslova, et al.
Pt 1): GIl80-8,
(2002) "Deficits in visceral pain and referred hyperalgesia in Nav1.8 (SNSIPN3)Neurosci 22(19): 8352-6), urinary incontinence
and visceral pain (See,Yoshimura,
aI. (2001) "The involvement of the tetrodotoxin-resistant
a rat model of visceral pain" J Neurosci
dysfunctions
null mice"
1
N., S. Seki, et
sodium channel Na(v)1.8 (PN3/SNS) in
21(21): 8690-6), as well as an array of psychiatry
such as anxiety and depression (See, Hurley, S. C. (2002) "Lamotrigine
its use in mood disorders" Ann Pharmacother
update and
36(5): 860-73).
[0003] Voltage gated Na channels comprise a gene family consisting of 9 different subtypes
(NaV1.1-NaV1.9).
functional
As shown in Table 1, these subtypes show tissue specific localization and
differences
(See, Goldin, A. L. (2001) "Resurgence
Annu Rev Physiol 63: 871-94).
Three members
of sodium channel research"
of the gene family (NaV1.8,
resistant to block by the well-known Na channel blocker ITX, demonstrating
1.9, 1.5) are
subtype specificity
within this gene family. Mutational analysis has identified glutamate 387 as a critical residue for
TIX; bW~g·(See.
Noda, M., H .. Suzuki;
et aI. (1989) "A .single point .mutation
on
confers.
tetrOdot6xiiifPI~ saxitoxin' insensitivity
the '$ocIitim channel II" FEBS Leti 259(1): 213-6}. . . .
.
.'
.....
.
[0004] Table 1 (Abbreviations: CNS central nervous system, PNS peripheral nervous sytem,
=
DRG
= dorsal
=
root ganglion, TG = Trigeminal ganglion):
Na
isoform
NaVl.1
NaV1.2
NaV1.3
Tissue
TIXIC50
Indications
CNS,PNS
soma of
neurons
eNS,high in
axons
CNS,
embryonic,
injured nerves
lOnM
Pain, Epilepsy,
neurodegeneration
lOnM
Neurodegeneration
Epilepsy
Pain
150M
-2-
..
WO 2005/042497
NaV1.4
NaV 1.5
NaVl.6
PCTlUS2004/036297
Skeletal
muscle
Heart
25nM
Myotonia
2~
Arrythmia,
long QT
Pain, movement disorders
eNS
6nM
widespread,
most abuntant
25nM
PNS,DRG,
terminals
neuroendocrine
PNS,small
>SOIJM
neurons in
DRG&TG
PNS, small
1~
neurons in
DRG&TG
NaVl.7
NaV1.8
NaV1.9
Pain, Neuroendocrine
disorders
Pain
Pain
[0005] In general, voltage-gated sodium channels (NaVs) are responsible for initiating the rapid
upstroke of action potentials in excitable tissue in nervous system, which transmit the electrical
signals that compose and encode nonnal and aberrant pain sensations.
Antagonists of NaV
channels can attenuate these pain signals and are useful for treating a variety of pain conditions,
including but not limited to acute, chronic, inflammatory, and neuropathic pain. Known NaV
antagonists, such as ITX, lidocaine (See, Mao, J. and L. L. Chen (2000) "Systemic lidocaine for
neuropathic pain relief' Pain 87(1): 7-17.) bupivacaine, phenytoin (See, Jensen, T. S. (2002)
uAltticonwisants in neUropathic pai~: 'ratio~aie and Cli~c~·.e~id~ns~'; Bur J Pain
61-8), larnotri~ine (See, Rozen, T.
D. (200i)
6 (Suppi A);
"AJitiepilepdc' drug~ in the inanagement of clu~t~r
headache and trigeminal neuralgia" Headache 41 Suppl 1: 325-32 and Jensen, T. S. (2002)
"Anticonvulsants in neuropathic pain: rationale and clinical evidence" Bur J Pain 6 (Suppl A):
61-8.), and carbamazepine (See, Backonja, M. M. (2002) "Use of anticonvulsants for treatment
of neuropathic pain" Neurology 59(5 SuppI2): S14-7), have been shown to be useful attenuating
.
.
pain in humans and animal models.
[0006] Hyperalgesia (extreme sensitivity to something painful) that develops in the presence of
tissue injury or inflammation reflects, at least in part, an increase in the excitability of highthreshold primary afferent neurons innervating the site of injury. Voltage sensitive sodium
channels activation is critical for the generation and propagation of neuronal action potentials.
There is a growing body of evidence indicating that modulation of NaV currents is an
-3-
2005/042497
- WO
~
PCTlUS2004/036297
endogenous mechanism used to control neuronal excitability (See, Goldin, A. L. (2001)
"Resurgence of sodium channel research" AnTIuRev Physiol 63: 871-94.). Several kinetically
and pharmacologically distinct voltage-gated sodium channels are found in dorsal root ganglion
(DRO) neurons. The TTX-resistant current is insensitive to micromolar concentrations of
tetrodotoxin, and displays slow activation and inactivation kinetics and a more depolarized
activation threshold when compared to other voltage-gated sodium channels. TIX-resistant
sodium currents are primarily restricted to a subpopulation of sensory neurons likely to be
involved in nociception. Specifically, TTX-resistant sodium currents are expressed almost
exclusively in neurons that have a small cell-body diameter; and give rise to small-diameter
slow-conducting axons and that are responsive to capsaicin. A large body of experimental
evidence demonstrates that TIX-resistant sodium channels are expressed on C-fibers and are
important in the transmission of nociceptive information to the spinal cord,
[0007] Intrathecal administration of antisense oligo-deoxynucleotides targeting a unique region
of the TI'X-resistant sodium channel (NaV1.8) resulted in a significant reduction in PO~induced .hyperalgesia (See, Khasar, S. G., M. S. Gold, et al. (1998) "A tetrodotoxin-resistant
sodium current mediates inflammatory pain in the rat" Neurosci Lett 256(1): 17-20). More
recently, a knockout mouse line was generated by Wood and coIIeagues, which lacks functional
NaV1.8. The mutation has an analgesic effect in tests assessing the animal's response to the
inflammatory agent carrageenan (See, Akopi an, A. N., V. Souslova, et al. (1999) ''The
~trodotoxin-resistant sodium channel SNS has a specialized function,~ pain pathways" ,Nat.", .
"NeUrQse:i
2(6):
.
..
54'l~8.i.Iil'addition,"defi~it in ~oth mechiUlo- ~d ihermareceptlon'were
.....
.
0'
..
",.
'.'
~bse~ed
.,'
in these animals. The analgesia shown by the Nav1.8 knockout mutants is consistent with
observations about the role of TTX-resistant currents in nociception.
[0008] Immunohistochemical, in-situ hybridization and in-vitro electrophysiology experiments
have all shown that the sodium channel NaV1.8 is selectively localized to the small sensory
·neurons of the dorsal root ganglion and trigeminal ganglion (See, Akopian,.A. N., L. Sivilotti, et
al. (1996) "A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons"
Nature 379(6562): 257-62,). The primary role of these neurons is the detection and transmission
of nociceptive stimuli. Antisense and immunohistochemical evidence also supports a role for
NaV1.8 in neuropathic pain (See, Lai, J., M. S. Gold, et aI. (2002) "Inhibition of neuropathic
pain by decreased expression of the tetrodotoxin-resistant sodium channel, NaV1.8" Pain 95(1-
-4-
W.,.?2005/042497
PCT/US2004/036297
2): 143-52, and Lai, 1., J. C. Hunter, et al. (2000) "Blockade
of neuropathic pain by antisense
targeting of tetrodotoxin- resistant sodium channels in sensory neurons" Methods Enzymol 314:
201-13.).
NaV1.8 protein is upregulated along uninjured C-fibers adjacent to the nerve injury.
Antisense
treatment
neuropathic
pain.
prevents
the redistribution
of NaV1.8
Taken together the gene-knockout
and antisense data support a role for
NaV1.8 in the detection and transmission of inflammatory
[0009] In neuropathic pain states there is a remodeling
along the nerve and reverses
and neuropathic pain.
of Na channel distribution and subtype.
In the injured nerve, expression of NaV1.8 and NaV1.9 are greatly reduced whereas expression
of the TTX sensitive subunit NaVl.3 is 5-10 fold upregulated
(1999) ''Plasticity
of sodium channel expression
injury model of neuropathic
(See, Dib-Hajj, S. D., J. Fjell, et al.
in DRG neurons in the chronic constriction
pain." Pain 83(3): 591-600.)
The timecourse
of the increase in
NaV1.3 parallels the appearance of allodynia in animal models subsequent to nerve injury.
biophysics
of the NaV1.3
inactivation
channel is distinctive
in that it shows very fast repriming
The
after
This allows for sustained rates of high firing as is
following an action potential.
often seen in the injured nerve (See, Cummins, T. R., F. Aglieco, et aI. (2001) "Nav1.3 sodium
channels: rapid repriming and slow closed-state inactivation
display quantitative differences after
in a mammalian cell line and in spinal sensory neurons" J Neurosci 21(16): 5952-
expression
61.). Na V1.3 is expressed in the central and peripheral
systems of man.
Na V1.9 is similar to
NaV1.8 as it is selectively localized to small sensory neurons of the dorsal root ganglion and
.. trigeminal ganglion (See. Fang. X; L. ·Djolihri,. et ale (2002). "The 'presepce and role· of the
- . . ..
.
. . '..
.'
- ...
tetrodotoxin-resistant
sodium
channel
Na(v)1.9
(NaN)
in
nociceptive
primary
afferent
neurons."
.
.,
.
.
.
.,
....
.
~.'
,"
'"
','
It has a slow rate of inactivation
J Neurosci
22(17): 7425-33.).
dependence
for activation (See, Dib-Hajj, S., J. A. Black, et al. (2002) "NaNlNav1.9:
channel
properties
with unique
properties"
allow NaV1.9 to playa
nociceptive neurons.
Trends
Neurosci
25(5):
role in establishing
253-9.).
and left-shifted
the resting membrane
potential
of
The resting membr-ane potential of Na V1.9 expressing cells is in the -55 to
is in large part due to the sustained
This depolarization
a sodium
These two biophysical
-50mV range compared to -65mV for most other peripheral and central neurons.
depolarization
voltage
low-level
activation
of NaV1.9 channels.
allows the neurons to more easily reach the threshold
potentials in response to nociceptive stimuli.
Compounds
This persistent
for firing action
that block the NaV1.9 channel may
play an important role in establishing the set point for detection of painful stimuli.
-5-
In chronic
WO
2005/042497
,>t
peT IUS2004/036297
pain states, nerve and nerve ending can become swollen and hypersensitive exhibiting high
frequency action potential firing with mild or even no stimulation.
These pathologic nerve
swellings are termed neuromas and the primary Na channels expressed in them are NaV1.8 and
NaV1.? (See, Kretschmer, T., L. T. Happel, et a1. (2002) "Accumulation of PN1 and PN3
sodium channels in painful human neuroma- evidence from immunocytochemistry" Acta
Neurochir CWien) 144(8): 803-10; discussion 810.). NaVl.6 and NaVl.7 are also expressed in
dorsal root ganglion neurons and contribute to the small TTX sensitive component seen in these
cells. NaV!.? in particular my therefore be a potential pain target in addition to it's role in
neuroendocrine excitability (See, Klugbauer, N., L. Lacinova, et al. (1995) "Structure and
functional expression of a new member of the tetrodotoxin- sensitive voltage-activated sodium
channel family from human neuroendocrine cells" Embo J 14(6): 1084-90).
[0010] NaV!.l (See, Sugawara, T., E. Mazaki-Miyazaki, et al. (2001) ''Navl.1 mutations cause
febrile seizures associated with afebrile partial seizures." Neurology 57(4): 703-5.) and NaVl.2
(See, Sugawara, T., Y. Tsurubuchi, et al. (2001) "A missense mutation of the Na+ channel alpha
IT subunit gene Na(v)l.2 in a patient with febrile and afebrile seizures causes channel
dysfunction" Proc Natl Acad Sci USA 98(11): 6384-9) have been linked to epilepsy conditions
including febrile seizures. There are over 9 genetic mutations in NaVl.1 associated with febrile
seizures (See, Meisler, M. H., J. A. Kearney, et a1. (2002) "Mutations of voltage-gated scdium
channels in movement disorders and epilepsy" Novartis Found Symp 241: 72-81)
.[OOU] Antagonists for NaVl.5 have ',been developed ~d, used to treat .cardiac arrhythmias.
A'
gei:ie,defect:in"Navi~5
that prod~eS
a'larger
no~naetivatitig
component 'to..."the curreni has' .be~n
,"
.
. .
..'
..
.
~ .'
.
.
.
linked to long QT in man and the orally available local anesthetic mexilitine has been used to
.
treat this condition (See, Wang, D. W., K. Yazawa, et al. (1997) "Pharmacological targeting of
long QT mutant sodium channels." J Clin Invest 99(7): 1714-20).
[0012] Several Na channel blockers are currently used or being tested in the clinic to treat
epilepsy ~
Moulard, a"and D. Bertrand (2002) ''Epilepsy and sodium channel·blo~keJ.'s~'
Expert Opin. Ther. Patents 12(1): 85-91.); acute (See, Wiffen, P., S. Collins, et al, (2000)
"Anticonvulsant drugs for acute and chronic pain" Cochrane Database Syst Rev 3), chronic (See,
Wiffen, P., S. Collins, et a1.(2000) "Anticonvulsant drugs for acute and chronic pain" Cochrane
Database Syst Rev 3, and Guay, D. R. (2001) "Adjunctive agents in the management of chronic
pain" Pharmacotherapy 21(9): 1070-81), inflammatory (See, Gold, M. S. (1999) "Tetrodotoxin-
- 6-
"
2005/042497
PCTlUS2004/036297
resistant Na+ currents and inflammatory hyperalgesia."
9), and
neuropathic
concentrations
pain (See,
Strichartz,
Proc Natl Acad Sci USA
G. R., Z. Zhou,
"Lamotrigine
Anaesthesiol
monotherapy
for control
of neuralgia
Scand 46(10): 1261-4); cardiac arrhythmias
(1996) ''Lidocaine
J Clin Invest 99(7):
Narasimhan
1714-20);
(1997) "Sodium channels
Phannacol
39: 47-98)
''Therapeutic
neuropathic
after nerve
Acta
(See, Taylor,
C. P. and L. S.
system diseases"
Adv
G. R., Z. Zhou, et al. (2002)
~Strichartz,
of local anaesthetics
section"
Circ Res 79(1): 103-8, and
and therapy of central nervous
and as anesthetics
concentrations
A., G. Rumpold S.eitlinger, et
targeting of long QT mutant sodium
neuroprotection
unveil the potential role of sodium channels in
pain." Novartis Found Symp 241: 189-201).
[0013] Y.arious animal models with clinical significance
sodium
"Therapeutic
(See, An, R. H., R. Bangalore, et al.
block of LQT-3 mutant human Na+ channels"
Wang, D. W., K. Yazawa, et al, (1997) "Pharmacological
channels"
et al. (2002)
of local anaesthetics unveil the potential role of sodium channels in neuropathic
pain" Novartis Found Symp 241: 189-201, and Sandner-Kiesling,
al. (2002)
96(14): 7645-
channel modulators
for numerous
different pain indications.
pain, see, Kohase, H., et al., Acta Anaesthesiol
(see, Kohase,
H., et al., Acta Anaesthesial
have been developed
E.g., malignant
Scand. 2004; 48(3):382-3;
Scand. 2004; 48(3):382-3);
et al., TQxicol Pathol. 2003; 31(6):619-24);
osteoarthritis
spinal stenosis (see, Takenobu,
chronic
femur cancer pain
non-malignant
bone pain (see, Ciocon, 1. O. et al., J Am Geriatr Soc. 1994; 42(6):593-6);
(see, Calvina, B. et a1., Behav Brain Res. 1987; 24(1):11-29);
for the study of
chronic
rheumatoid arthritis
(see, Guzman, R. E.,
Y. et al" .JNeurosci
'. MethOdS., 2001; 104(2):191-8);
Neui:opa$ic low back pain (~ee: Iiin~, R., et' al,,. Pain Med,
.
,"..
.
.
2002; 3(4):361-5; Massie, J. B., et al., J Neurosci Methods, 2004; 137(2):283-9;
'.
.
neuropathic
."
low back pain (see, Hines, R., et al., Pain Med. 2002; 3(4):361-5; Massie, J. B., et
al., J Neurosci Methods. 2004; 137(2):283-9); myofascial pain syndrome (see, Dalpiaz & Dodds,
J Pain Palliat Care Pharmacother. 2002; 16(1):99-104; Sluka KA et al., Muscle Nerve. 2001;
24(1):37-46);
fibromyalgia (see, Bennet &.Tai, Int J CUn Pharmacol Res. 1995;15(3):115-9);
temporomandibular
joint pain (see, Ime H, Ren K, Brain Res Mol Brain Res. 1999; 67(1):87-97);
chr~mic visceral pain, including, abdominal (see, AI-Chaer, E. D., et al., Gastroenterology.
119(5):1276-85);
6); pancreatic
pelvic/perineal pain, (see, Wesselmann
(see, Vera-Portocarrero,
et al., Neurosci Lett. 1998; 246(2):73-
L. B., et al., Anesthesiology.
-7-
2000;
2003; 98(2):474-84);
.
..
PCT/uS2004/036297
WO 2005/042497
IDS pain (see, Verne, G. N., et al., Pain. 2003; 105(1-2):223-30; La JH et aI., World
2003; 9(12):2791-5); chronic headache pain (see, Willimas & Stark, Cephalalgia.
Gastroenterol.
2003; 23(10):963-71);
migraine (see, Yamamura,
H., et al., J Neurophysiol. 1999; 81(2):479-93);
. tension headache, including, cluster headaches (see, Costa, A., et al., Cephalalgia. 2000;
20(2):85-91);
Neurology.
chronic neuropathic pain, including, post-herpetic neuralgia (see, Attal, N., et al.,
2004; 62(2):218-25; Kim & Chung 1992, Pain 50:355); diabetic neuropathy (see,
Beidoun A et al., Clin J Pain. 2004; 20(3):174-8; Courteix, C., et al., Pain. 1993; 53(1):81-8);
IDV- associated neuropathy (see, Portegies & Rosenberg, Ned Tijdschr Geneeskd. 2001;
145(15):731-5;
Joseph EK et al., Pain. 2004; 107(1-2):147-58;
Oh, S. B., et aI., J Neurosci. 2001;
trigeminal neuralgia (see, Sato, J., et aI., Oral Surg Oral Med Oral Pathol Oral
21(14):5027-35);
Radiol Endod. 2004; 97(1):18-22; Imamura Y et al., Exp Brain Res. 1997; 116(1):97-103);
Charcot-Marie
Tooth neuropathy
(see, Sereda, M., et al., Neuron. 1996; 16(5): 1049-60);
hereditary sensory neuropathies (see, Lee, M. J., et aI., Hum Mol Genet. 2003; 12(15):1917-25);
peripheral nerve injury (see, Attal, N., et al., Neurology. 2004; 62(2):218-25; Kim & Chung
1992, Pain 50:355; Bennett & Xie, 1988, Pain 33:87; Decostered, I. & Woolf, C. J., 2000, Pain
87:149; Shir, Y. & Seltzer, Z. 1990; Neurosci Lett 115:62); painful neuromas (see, Nahabedian
& Johnson, Ann Plast Surg. 2001; 46(1):15-22; Devor & Raber, Behav Neural BioI. 1983;
ectopic proximal and distal discharges (see, Liu, X. et aI., Brain Res. 2001;
37(2):276-83);
900(1): 119-27); radiculopathy
N et a!.! Spine .. 1998; 23(8):877-85); chemotherapy induced neuropathic'pain
Hayashi
K.
0.,
. .
(see, Devers & Galer, (see, Clin J Pain. 2000; 16(3):205-8;
et al:"~eurosciexice.
.
"
1996; 73(1):25~,.65);· radiotherapy-~duced
neuropathic pain;
..,....
....
..
pain (see, Devers & Galer, Clin J Pain. 2000; 16(3):205-8); central pain
.,
post-mastectomy
(see, Aley;
.'
(Cabana, A, et aI., Anesth Analg. 2004; 98(6):1581-4),
-
spinal cord injury pain (see, Hains, B. C.,
et al., Exp Neural. 2000; 164(2):426-37); post-stroke pain; thalamic pain
(see, LaBuda, C. J., et
al., Neurosci LeU. 2000; 290(1):79-83); complex regional pain syndrome (see, Wallace, M. S., et
aI., Anesthesiology.
2000; 92(1):75-83; Xantos D et al., J Pain. 2004; 5(3 SuppI2):Sl);
pain (see, Weber, W.
phanton
E., Ned Tijdschr Geneeskd. 2001; 145(17):813-7; Levitt & Heyback, Pain.
1981; 10(1):67-73); intractable pain (see, Yokoyama, M., et al., Can J Anaesth. 2002; 49(8):8103); acute pain, acute post-operative pain (see, Koppert, W., et al., Anesth Analg. 2004;
98(4):1050-5;
Brennan, T. J., et al., Pain. 1996; 64(3):493-501);
pain (see, Gotoh, S., et aI., Ann RheumDis.
acute musculoskeletal
1993; 52(11):817-22);
-8-
pain; joint
mechanical low back pain
W~005/042497
PCT/US2004/036297
(see, Kehl, L. J., et al., Pain. 2000; 85(3):333-43); neck pain; tendonitis; injuryfexercise pain
(see, Sesay, M., et a1., Can J Anaesth. 2002; 49(2):137-43);
acute visceral pain, including,
abdominal pain; pyelonephritis;
intestinal obstruction; hernias; etc
(see, Giambernardino,
appendicitis; cholecystitis;
M. A., et aI., Pain. 1995; 61(3):459-69);
chest pain, including, cardiac
Pain (see, Vergona, R. A., et aI., Life Sci. 1984; 35(18):1877-84);
acute obstetric
pelvic pain, renal colic pain,
pain, including, labor pain (see, Segal, S., et al., Anesth Analg. 1998; 87(4):864-
9); cesarean section pain; acute inflammatory, burn and trauma pain; acute intermittent pain,
including, endometriosis
(see, Cason, A. M., et al.,Horm Behav. 2003; 44(2):123-31);
acute herpes zoster pain; sickle cell anemia; acute pancreatitis
2000; 119(5):1373-81);
breakthrough
pain; orofacial pain, including,
(see, Nusstein, J., et al., J Ended. 1998; 24(7):487-91;
6(1):55-67);
multiple
162(2):162-8);
leprosy
Haglund
Venereol.
disease
1999; 24(3):240-1);
syndrome;
2003;
2002;196(2):183-90);
(see, Devillers
phlebitic pain; Guillain-Barre
erythromelalgia
Bladder
& Kanazawa,
pain (see, Legroux-Crespel,
Fabry's
disease
and urogenital
pain
disease,
aI., Urology. 2003; 61(3):664-70);
E., et al., Ann Dermatol
(see, Germain,
including,
painful" bladder syndrome
.' NeU:rbs~i. 20~1; '21(2~):'8690-6);' int~~titiai' cyc~ti·s.
& Oranje, Clin Exp
pain; painful legs and moving toes;
D. P., J Soc BioI.
urinary incontinence
Berggren, T., et al., J Urol. 1993; 150(5 Pt 1): 1540-3); hyperactivity
et
J Neurol Sci. 1999;
(see, Greene B, Curr Med Res Opin. 2003; 19(4):272-7);
pain; adiposis dolorosa
130(4):429-33);
sinusitis pain, dental pain
J. J., et al., Eur J Pain. 2002;
Chidiac,
(MS) pain (see, Sakurai
pain in depression
pain; behcet's
Dermatol.
sclerosis
(see, Toma, H; Gastroenterology.
(see,
bladder (see, Chuang, Y. C.,
(see, Yoshimura, N., et: at, J
6¢) :(s~e.' Gia:nnak6poulos&
Arch Ital Urol Nefrol AndroI. 1992; 64(4):337-9; Boucher, M., et al., JUral.
8); and prostatitis (see, Mayersak, J. S., Int Sorgo 1998; 83(4):347-9;
.~.~pilo~ai~s,
2000; 164(1):203-
Keith, I. M., et al., J Urol.
2001; 166(1):323-8).
[0014] Voltage-gated calcium channels are membrane-spanning,
open in response to membrane depolarization,
multi-subunit proteins that
allowing Ca entry from the extrace1hl1ar milieu.
Calcium channels were initially classified based on the time and voltage-dependence
opening and on the sensitivity to pharmacological
of channel
block. The categories were low-voltage
activated (primarily T-type) and high-voltage activated (L,N,P,Q or R-type). This classification
scheme was replaced by a nomenclature based upon the molecular subunit composition, as
summarized in Table I (Bockerman, G. H., et. al. (1997) Annu. Rev. Pharmacol. Toxicol. 37:
-9-
peTIUS2004/036297
WQi005/042497
361-96; Striessnig, J. (1999) Cell. Physio!. Biochem. 9: 242-69). There are four primary subunit
types that make up calcium channels - al,
1120, ~
and functional diversity of voltage-activated
and
y (See, e.g., De Waard et al. Structural
calcium channels. In Ion Channels, (ed. T.
Narahashi) 41-87, (plenum Press, New York, 1996». The
of the pharmacological
UI
subunit is the primary determinant
properties and contains the channel pore and voltage sensor (Hockerman,
G. H., et. al. (1997) AnTIu.Rev. Pharmacal. Toxicol. 37: 361-96; Striessnig.
Physio!. Biochem. 9: 242-69). Ten isoforrns of the
III
J. (1999) Cell.
subunit are known, as indicated in Table 1.
The a20 subunit consists of two disulfide linked subunits, a2, which is primarily extracellular and
a transmembrane
0 subunit. Four isoforms of a2~ are known, a20-1, a20-2, 1128-3 and U20-4. The
~ subunit is a non-glycosylated
are known, termed
biochemically
~l
to
~4.
cytoplasmic protein that binds to the al subunit. Four isoforrns
The'l subunit is a transmembrane protein that has been
isolated as a component of Cavl and Cay2 channels. At least 8 isoforms are
known (Y1 to Y8) (Kang, M.G. and K. P. Campbell (2003) 1. BioI. Chern. 278: 21315-8). The
nomenclature
for voltage-gated calcium channels is based upon the content of the al subunit, as
indicated in Table I. Each type of
<Xl
subunit can associate with a variety of ~'
subunits, so that each Cay type corresponds to many different combinations
<X28
or y
of subunits.
Cay Nomenclature
al subunit
Pharmacological name
Cavl.l
als
L-type
Cav1.2
ale
L-type
Cay1.3
·CavI.4
...
..
....
aID
"
"
~7trPe .
.,
aiF'
Cav2.1
alA
P-orQ-type
Cav2.2
alB
N-type
Cay2.3
alE
R-type
Cay3.1
alG
T-type
Cav3.2
. aIR
T-type
Cav3.3
an
T-type
. [0016] Cay2 currents are found almost exclusively in the central and peripheral nervous system
and in neuroendocrine
cells and constitute the predominant forms of presynaptic
voltage-gated
calcium current. Presynaptic action potentials cause channel opening and neurotransmitter
- 10-
..
WO 10051042497
PCTlUS2004/036297
release is steeply dependent upon the subsequent calcium entry. Thus, Cav2 channels playa
central role in mediating neurotransmitter
release.
[0016] Cay2.1 and Cay2.2 contain high affinity binding sites for the peptide toxins D-conotoxinMVITC and D-conotoxin-GVIA,
respectively, and these peptides have been used to determine
the distribution and function of each channel type. Cav2.2 is highly expressed at the presynaptic
nerve terminals of neurons from the dorsal root ganglion and neurons of lamina I and ITof the
dorsal horn (Westenbroek,
R. E., et a1. (1998) J. Neurosci. 18: 6319-30; Cizkova, D, et a1. (2002)
Exp. Brain Res. 147: 456-63).
Cav2.2 channels are also found in presynaptic terminals between
second and third order intemeurons in the spinal cord. Both sites of neurotransmission
are very
important in relaying pain information to the brain.
[0017] Pain can be roughly divided into three different types: acute, inflammatory, and
neuropathic.
Acute pain serves an important protective function in keeping the organism safe
from stimuli that may produce tissue damage.
Severe thennal, mechanical, or chemical inputs
have the potential to cause severe damage to the organism if unheeded.
quickly remove the individual from the damaging environment.
generally is short lasting and intense. Inflammatory
Acute pain serves to
Acute pain by its very nature
pain, on the other hand, may last for much
longer periods of time and its intensity is more graded. Inflammation
may occur for many
reasons including tissue damage, autoimmune response, and pathogen invasion.
pain is mediated by a variety of agents that are released during inflammation,
., substance P, histamines; acid, prostaglandin, ·bradykinin, CGRP, 'eyto~es,
agents (julius, D ..and
.'
".
is neuropathic
A.l..
Basba:~
....
(2001)
including
ATP, and other.
Nature 413 (6852):
.....203-iO)... Th~
~
--,-
Inflammatory
third
..
"
c1as's of Pain. .
.
".
and involves nerve damage arising from nerve injury or viral infection and results
in reorganization
of neuronal proteins and circuits yielding a pathologic "sensitized"
state that
can produce chronic pain lasting for years. This type of pain provides no adaptive benefit and is
particularly difficult to treat with existing therapies .
.[0018]. Pain, particnlarly neuropathic and intractable pain is ~ large unmet medical need.
Millions of individuals suffer from severe pain that is not well controlled by current therapeutics.
The current drugs used to treat pain include NSAIDS, COX-2 inhibitors, opioids, tricyclic
antidepressants,
and anticonvulsants.
Neuropathic
pain has been particularly difficult to treat as
it does not respond well to opioids until high doses are reached.
Gabapentin is currently the most
widely used therapeutic for the treatment of neuropathic pain, although it works in only 60% of
-11-
"
¥) 2005/042497
PCTlUS2004/036297
..
patients and has modest efficacy. The drug is generally safe, although sedation is an issue at
higher
doses.
[0019] Validation of Cav2.2 as a target for the treatment of neuropathic pain is provided by
studies with ziconotide (also known as D-conotoxin-MVIIA),
a selective peptide blocker of this
channel (Bowersox, 8.S., et al. (1996) 1. Pharmacol. Exp. Ther. 279: 1243-9; Jain, K.K. (2000)
Exp. Opin. Invest. Drugs 9: 2403-10; Vanegas, H. and H. Schaible (2000) Pain 85: 9-18). In
man, intrathecal infusion of Ziconotide is effective for the treatment of intractable pain, cancer
pain, opioid resistant pain, and neuropathic pain. The toxin has an 85% success rate for the
treatment of pain in humans with a greater potency than morphine.
An orally available
antagonist of Cav2.2 should have similar efficacy without the need for intrathecal infusion.
Cav2.1 and Cav2.3
are also in neurons of nociceptive pathways and antagonists of these .
channels could be used to treat pain.
[00201 Antagonists of Cav2.1, Cav2.2 or Cav2.3 should also be useful for treating other
pathologies of the central nervous system that apparently involve excessive calcium entry.
Cerebral ischaemia
of neurons.
and stroke are associated with excessive calcium entry due to depolarization
The Cav2.2 antagonist ziconotide is effective in reducing infarct size in a focal
ischemia model using laboratory animals, suggesting that Cav2.2 antagonists could be used for
the treatment of stroke. Likewise, reducing excessive calcium influx into neurons may be useful
for the treatment of epilepsy, traumatic brain injury, Alzheimer's disease, multi-infarct dementia
and other classes of demen.tia,.amyotrophic
lateral.sclerosis,
by poison pr other toxic substances.
."
. '.
.
[0021] Cav2.2 also mediates release of neurotransmitters
amnesia, or neuronal damage caused
from neurons of the sympathetic
nervous system and antagonists could be used to treat cardiovascular
diseases such as
hypertension, cardiac arrhythmia, angina pectoris, myocardial infarction, and congestive heart
failure .
.[()022] Howeyer, as .described above, the efficacy of.currently used sodium channel blockers and
calcium channel blockers for the disease states described above has been to a large extent limited
by a number of side effects. These side effects include various CNS disturbances such as blurred
vision, dizziness, nausea, and sedation as well more potentially life threatening cardiac
arrhythmias and cardiac failure. Accordingly, there remains a need to develop additional Na
- 12-
WO 2005/042497
PCTlUS2004/036297
"'-
channel antagonists, and Ca channel antagonists preferably those with higher potency and fewer
side effects.
SUMN.lARY OF THE INVENTION
[0023]
It has now been found that compounds
acceptable
compositions
thereof,
of this invention,
are useful as inhibitors
These compounds have the structure of Formula
and pharmaceutically
of voltage-gated
sodium channels.
I:
I
acceptable salt thereof, wherein RA, Z, Y, r, and Ware as defined below.
or a pharmaceutically
These compounds
and pharmaceutically
acceptable
compositions
are useful for
treating or lessening the severity of a variety of diseases, disorders, or conditions, including, but
not limited to, acute, chronic, neuropathic, or inflammatory
pain such as femur cancer pain; non-
malignant chronic bone pain; rheumatoid arthritis; osteoarthritis;
back
.....
pain;
neuropathic
low
back
pain;
myofascial
spinal stenosis; neuropathic low
pain
syndrome;
fibromyalgia;
temporomandibular
joint pain; chronic visceral pain, including, abdominal; pancreatic; IDS pain;
chroI).ic headache
pain; migraine;
:
•
.neuropathic
'neuropathy;
':
••••
tensiop headache,
••
pain, including"
trlgeniin;U
o'
•
post-herpetic. neuralgia;
neuralii~;'
including,
cluster hea,dachel); chronic
..'
•
diabetic
••
•
neuropathy;
cJiai-~ot~M~e : Tooth . 'neuropathy;
,"
••
HIV - ' ,associated
'hereditary
sensory
neuropathies; peripheral nerve injury; painful neuromas;
ectopic proximal and distal discharges; radiculopathy; chemotherapy
radiotherapy-induced
neuropathic pain; post-mastectomy
induced neuropathic pain;
pain; central pain; spinal cord injury
:pain; post~stro,~e pain; thalamic p~n; complex regional pain syndro:me; phanton 1?ain;intr~ctabl.e,
pain; acute pain, acute post-operative
pain; acute musculoskeletal
pain; joint pain; mechanical
low back pain; neck pain; tendonitis; injury/exercise pain; acute visceral pain, including,
abdominal pain; pyelonephritis;
appendicitis; cholecystitis;
intestinal obstruction; hernias; etc;
chest pain, including, cardiac Pain; pelvic pain, renal colic pain, acute obstetric pain, including,
labor pain; cesarean section pain; acute inflammatory, burn and trauma pain; acute intennittent
-13 -
· "Y-~2005/042497
PCTIUS2004/036297
pain, including, endometriosis;
acute herpes zoster pain; sickle cell anemia; acute pancreatitis;
breakthrough pain; orofacial pain, including, sinusitis pain, dental pain; multiple sclerosis (MS)
pain; pain in depression; leprosy pain; behcet's disease pain; adiposis dolorosa; phlebitic pain;
Guillain-Barre pain; painful legs and moving toes; Haglund syndrome; erythromelalgia
pain;
Fabry's disease pain; bladder and urogenital disease, including, urinary incontinence;
hyperactivity bladder; painful bladder syndrome; interstitial cyctitis (Ie); or prostatitis, arthritis,
migrane, cluster headaches, trigeminal neuralgia, herpetic neuralgia, general neuralgias, epilepsy
or epilepsy conditions, neurodegenerative
disorders, psychiatric disorders such as anxiety and
depression, myotonia, arrythmia, movement disorders, neuroendocrine
disorders, ataxia, multiple
sclerosis, irritable bowel syndrome, and incontinence.
DETAILED DESCRIPTION OF THE INVENTION
1. General Description of Compounds of the Invention:
[0024]
The present invention provides a compound of Formula I:
y
o=
I .
z
(RA)r-,
)-w
#N
.'I. '
ot a,pharmac~~tically
acceptable salt thereof, w~ere~:
r is 0 to 4;
Z is 0, N or CH;
Y and W are independently selected from hydrogen, Formula Ia:
-,-A-V-
Ia
wherein:
- 14-
~
2005/042497
PCTlUS2004/036297
, or
wherein:
T is a bond or a CI-6 straight or branched aliphatic chain wherein a methylene unit
ofT is optionally replaced by a C3.s cycloaliphatic
group;
U is -CHz- or -CHz-CHz-;
X is N-CI-4aIkyl, NH, 0, S, S(O), or SOz; and
each occurrence ofRe is independently
M_Rx; wherein:
M is a bond or is a CI-6 alkylidene chain wherein up to two non-adjacent
methylene units of M
are optionally replaced by C(O), COz,
C(O)C(O), C(O)NR, OC(O)NR, NRNR, NRNRC(O),
NRC(O),
NRCOz, NRC(O)NR, S(O), SOz, NRSOz, SOzNR, NRSOzNR,
0, S,
orNR, and
X
R is R', halogen, NOz, or CN; wherein:
each occurrence ofR' is independently
selected from hydrogen or an
optionally substituted group selected from C1-S aliphatic, CG-IO aryl,
a heteroaryl ring having 5-10 ring atoms, or a heterocyclyl ring
, 'having 3-10 ring atoms, or Rand R' ..taken together with the ....
.' ato:t:Dcs)t,oWhicq they ar~ .bOl~~i or
two
~ccurrenc~~ 9~R; t*~n,
together with the atom(s) to which they are bound, form a 5-8
membered cycloalkyl, heterocyclyl,
0-3 heteroatoms independently
aryl, or heteroaryl ring having
selected from nitrogen, oxygen, or
sulfur;
V is a bond, -C(O)-, or -S(0)z-; .
Q is a bond or a Cl-4 alkylidene chain wherein up to two non-adjacent
methylene units of
Q are optionally replaced by -0-, -NH-, or -S-;
mis 0 or 1;
Ring E is
C6.10
aryl, a 5-10 membered heteroaryl ring having 1-4 heteroatoms
independently
selected from nitrogen, oxygen, or sulfur, or a 3-10 membered
- 15 -
~
2005/042497
PCT/uS2004/036297
heterocyclyl ring having 1-4 heteroatoms independently
selected from nitrogen,
oxygen,orsuffur;and
s is 0 to 8;
or Formula
Ib:
Ib
wherein:
D is -Cl_6alkyl- or a bond; and
tis 0 to 5;
each occurrence ofR is independently
selected from hydrogen or an optionally substituted Cl-6
aliphatic group; and
each occurrence ofRA, RB andRD are independently selected fromRl,
R2, R3, R4, orR5,
wherein:
Rl is oxo, R6 or (Cl-4aliphatic)n-J, wherein:
n is 0 or 1;
J is halo, CN, N02, CF3' OCF3' OH, SR6, S(O)R6, S02R6, NH2, NHR6, N(R6)z,
NR6R8, C(O)OH, C(O)OR6 or OR6; or:
~oR io~ adj~c.ent ~g
. 'ethylenedioxy;
a~oms, taken togeth~r, form 1,2-m~thYlen~~oxy
or 1,2-
'. .
R2 is Cl_6aliphatic, optionally substituted with 'up to two substituents independently
selected from R 1, R4, or R 5;
R3 is C3_scycloaIiphatic, C6-JOaryl, a 5-10 membered heteroaryl ring having 1-4
··heteroatoms independently
selected from nitrogen, oxygen\ or sulfur, or a 3-10.
membered heterocyclyl ring having 1-4 heteroatoms independently
3
selected from
nitrogen, oxygen, or sulfur, wherein R is optionally substituted with up to three
substituents independently
selected from Rl, R2, R4 or R5;
- 16 -
...
2005/042497
PCTruS20041036297
R4 is OR5, OR6, OC(O)R6, OC(O)R5, OC(O)OR6, OC(O)OR5, OC(O)N(R6h,
OC(O)N(R5)2,
SOzN(R6h,
OC(O)N(R6R5),
S02N(R5h,
C(O)R6, qO)OR6,
C(O)N(OR5)R6,
SR6, SR5, S(O)R6, S(O)RS, SOZR6, S02RS,
S02NR5R6,
S03R6,
S03RS, C(O)R5, C(O)OR5,
C(O)N(R6)z, C(O)N(R5h,
C(O)N(OR6)R5,
C(O)N(R5R6),
C(O)N(ORS)R5,
C(O)N(OR6)R6,
C(NOR6)R6,
C(NOR6)R5,
C(NOR5)R6, C(NOR5)R5, N(R6)Z, N(R5)2, N(R5R6), NR5C(O)R5, NR6C(O)R6,
NR6C(O)R5,
NR6C(O)OR6,
NR5C(O)OR6,
NR6C(O)OR5,
NR6C(O)N(R6h,
NR6C(O)NR5R6,
NR6C(O)N(R5)z,
NR5C(O)NR5R6,
NRSC(O)N(R5)z,
NR6S0ZR6,
z
NR6S02NR5R6,
NR5S02N(R5)Z'
N(OR6)R6, N(OR6)R5, N(OR5)R5,
NR6S02N(R5)z,
NR5S0ZR5,
NR5S02NR5R6,
or N(OR5)R6;
aryl, a 5-10 membered heteroaryl ring having 1-4
C6-JO
heteroatoms independently
NR5C(O)N(R6)Z,
NR6S0ZRS,
NR6S0 N(R6h,
RS is a C3_scycloaliphatic,
NR5C(O)OR5,
selected from nitrogen, oxygen, or sulfur, or a 3-10
membered heterocyclyl ring having 1-4 heteroatoms
independently
selected from
nitrogen, oxygen, or sulfur, wherein R5 is optionally substituted with up to three R1
substituents;
R6 is'R 0J?tioria1Iy'substi~ted with R:, ,!herein:
. R7'i~a C3~cycloaliphatic,
.
aryl; a 5-i.·O·me~bered hei~ro~l
C6-10
heteroatoms independently
.
ring having 14"
selected from nitrogen, oxygen, or sulfur, or a 3-10
membered heterocyclyl ring having 1-4 heteroatoms
independently
selected from
nitrogen, oxygen, or sulfur, wherein R7 is optionally substituted with up to two
substituents independently
ethylenedioxy,
selected from R, 1,Z-methylenedioxy,
1,Z-
or (CH2)n-G, wherein Gis seiected rrom halo, CN, NOz,
OCF3, OH, S-aliphatic, S(O)-aliphatic,
N(aliphatic)Z, N(aliphatic)R8,
SOZ-aliphatic,
COOH, C(O)O(-aliphatic,
CF3,
NHZ' N-aliphatic,
or O-aliphatic; and
R 8 is an amino protecting group;
provided that only one of Y and W is formula Ia or
- 17-
Ib and the other of Y and W is hydrogen.
WO 2005/042497
PCT/US20041036297
-/
In certain other embodiments, for compounds of Formula I as described generally
[0025]
above and herein:
a) when Z is N, W is Formula la, A is -C2alkyl-NH-,
V is -CCO)-, Q is -Cjalkyl-O-,
and
A
Ring E is phenyl, then R is not hydrogen, -CI, -Br, Cj-4alkyl, methoxy, or nitro, either
singly or in combination;
b) when Z is N, W is Formula la, A is -C2alkyl-NH-,
V is -eCO)-, Q is a bond, and Ring
E is phenyl, then RA is not hydrogen, -CI, -Br, CHalkyl, methoxy, or nitro, either singly
or in combination;
c) when Z is N, W is Formula la, A is -e3alkyl-NH-,
V is -CCO)-,
Q is a bond, and Ring
A
E is phenyl, then R is not 4-amino, or 4-methoxycarbonyl;
d) when Z is N, W is Formula la, A is -C3alkyl-NH-,
V is -CCO)-,' and Q is a bond, then
Ring E is not -2(2,3-dihydro-benzo[lAJdioxine);
e) when Z is N, W is Formula la, A is -C3alkyl-NH-,
-Cjalkyl-O-,
f)
V is -CCO)-, and Q is a
then Ring E is not -6(4-dirnethyl-2H-chromen-2-one);
when Z is N, W is Formula
la, A is -C2alkyl-NH-,
-C2alkyl-O-, then Ring E is not unsubstituted
V is -C(O)-,
and Q is a
phenyl;
g) when Z is N, W is Formula la, A is -e2alkyl-NH-,
V is -CCO)-, and B is a bond, then
Ring E is not unsubstituted thienyl;
h)
when Z is N, W is Formula la, A is -e2alkyl-NH-,
V is -C(O)-,
A
'.' . -C1alkyl-0-,. and Ring E is phenyl, then R is not phenyl at the 4 position;'
and Q is a
.'
i) ~he~·~.i~ N, Wis~0n.nula)~,.A.is·~2alkYl:-N:Eh
.
.
then Ring E is not 2-isoindoline-l ,3-dione;
Vis-C(Oh.and
Q.~~:a-:C2~YI,.
j) when Z is N, W is Formula la, A is -C2alkyl-NH-,
V is -C(O)-, and Q is a
A
-C2alkyl-O-, and Ring E is phenyl, then R is not phenyl at the 4 position; and
k) when Z is N, W is Formula la, A is -e2alkyl-NH-,
V is -e(O)-,
and Q is a bond, then
Ring E is not unsubstituted adamantyI.
[0026]
According
to another embodiment,
formula la, as defined generally above, wherein:
·18 -
the present invention provides
a compound
of
PCTlUS2004/036297
WO 2005/042497
I"qR
C
>(
~
N-~
(a) when Z is N, Y is hydrogen, W is formula la, A is
and V and Q are each a
0,
bond, then:
(i) when r is 1 and RA is methyl in the C-5 or C-6 position of the benzimidazole ring, then
E is not:
unsubstitued phenyl;
phenyl substituted in the ortho position with methyl, OMe, or OEt; or
phenyl substituted in the para position with OMe or methyl; and
(ii) when r is 0, then E is not:
unsubstituted phenyl;
unsubstituted naphthyl;
phenyl substituted in the para position with OEt, Br, OR, or OMe;
phenyl substituted in the meta position with chiaro; or
phenyl substituted in the ortho position with methyl;
(b) when Z is N, Y is hydrogen, W is formula la, Q is -NHCHz-, r is 0, and V is ceO), then:
cl~i
(i) when A is -CHzCH~-,
th~n E is not
.'
- 0
~;
~0M.
~.
(ii) when A is -CHzNH-, then E is not
""'('CI
CI;
and
and
(c) when Z is C, W is hydrogen, Y is formula la, r is 0, A is -eH2CHzNH-,
-CH20- and E is phenyl, then:
(i) s is not 0;
.
(ii) when s is 1, RB is not:
.
- 19-
V is C(O),
Q is
PCTlUS2004/036297
WO 2005/042497
unsubstituted
phenyl,
chloro,
OMe,
methyl,
bromo,
~
\-~''N'/l
in the para position;
~o
cyano or OMe in the ortho position; or
methyl in the meta position;
(iii) when s is 2, RB is not dichloro in the ortho/para positions; and
(iv) when s is 3, RB is not 2,3,4-trimethyoxy
In certain other embodiments,
[0027]
or 2,4,5-trichloro.
for cpmpounds
of Formula I as described generally
above and herein:
a) when Z is N, W is Formula la, A is -CH2CH2NH-, V is -C(O)-,
Q is -CH20-, and
Ring E is phenyl, then RA is not -Cl, -Br, Cl-4aIkyl, methoxy, or nitro, either singly or in
combination;
b) when Z is N, W is Formula la, A is -CH2CHzNH-, V is -C(O)-,
A
E is phenyl, then R
Q is a bond, and Ring
is not -Cl, -Br, CI-4alkyl, methoxy, or nitro, either singly or in
combination;
c) when Z is N, W is Formula la, A is -CH2CH2CH2NH-, V is -C(O)-, Q is a bond, and
Ring E is phenyl, then RA is not 4-arnino, or 4-methoxycarbonyl;
d) wh~n Z is N,
'w is Forin~lata;'A
is ~2CH2CH2NH-,
V is :::C(O)-, arid
",the~ Ri~giHs not -2(2,3-diliyciro~b~~o[i,4]~oxine);
"
e) when Z is N, W is Formula la, A is -CH2CH2CH2NH-,
,;'"
Q' is a bond:'
,,;,"
V is -C(O)-, and Q is a
-CH20-, then Ring E is not -6(4-dimethyl-2H-chromen-2-one);
f)
when Z is N, W is Formula
la, A is -CH2CH~-,
V is -C(O)-,
and Q is a
-CH2CH20-, then Ring E is not unsubstituted phenyl;
g)' when Z is N, W is'Formula la, A is -CH2CH2NH-, V is'-C(O)-,
and Q is a bond, then
Ring E is not unsubstituted thienyl;
h) when Z is N, W is Formula la, A is -CH2CH2NH-,
V is -C(O)-,
and Q is a -CH20-,
and Ring E is phenyl, then RA is not phenyl at the 4 position;
i) when Z is N, W is Formula la, A is -CH2CH2NH-, V is -C(O)-, and Q is -CH2CH2-,
then Ring E is not 2-isoindoline-l,3-dione;
- 20-
'
wo
2005/042497
.
..:
PCTlUS2004/036297
j) when Z is N, W is Formula la, A is -CHzCHzNH-,
V is -C(O)-, and Q is -CH2CHzO-,
and Ring E is phenyl, then RA is not phenyl at the 4 position; and
k) when Z is N, W is Formula la, A is -CHzCHzNH-,
./
V is -C(O)-, and Q is a bond, then
Ring E is not unsubstituted adamantyl.
1) when Z is N, W is Formula la, A is -CHzCHzNH-,
V is -C(O)-, Q is -CH2CH20-,
and
Ring E is phenyl, then RB is not -Cl, -Br, CI_4alkyl, methoxy, unsubstituted phenyl,
-C(CH3hphenyl,
or nitro, either singly or in combination;
m) when Z is N, W is Formula la, A is -CH2CH2NH-,
V is -C(O)-, Q is -CH=CH2-, and
B
Ring E is phenyl, then R is not -Cl in the ortho position; and
n) when Z is N, W is Formula la, A is -CHzCH2NH-,
V is -802-, Q is a bond, and Ring
B
E is phenyl, then R is not chloro.
[0028]
Another
lessening
the severity
neuropathic,
embodiment
of a disease,
or inflammatory
pain; rheumatoid
of the present
disorder,
invention
or condition
headache,
heIJletic neuralgia;
Charcot-Marie
p~~l
pancreatic;
including, 'cluster
pib~ar
temporomandibular
chronic
neuropathic
illV - associated
hereditary
pain; radiotherapy-induced
sensory
neuropathies;
neuropathic
obstruction;
obstetric
pain; migraine;
trigeminal
post-
neuralgia;
peripheral 'nerve .injury; ,
pain; thalamic
pain; post-mastectomy
pain; complex
joint pain; mechanical low back pain; neck pain; tendonitis;
abdominal
joint pain; chronic
pain, including,
neuropathy;
phanton pain; intractable pain; acute pain, acute post-operative
including"
chronic bone
and dist~ discharge~,; r8:clic~opathy;, chenio~~taPY indu~e.d
spinal cord injury pain; post-stroke
pain,
from acute, chronic,
IBS pain; chronic headache
headaches;
neuropathy;
Tooth neuropathy;
n<?m:omas; ~tOpic
neuropathic
abdominal;
diabetic
selected
or
spinal stenosis; neuropathic low back pain; neuropathic
low back pain; myofascial pain syndrome; fibromyalgia;
tension
a method of treating
pain, including femur cancer pain; non-malignant
arthritis; osteoarthritis;
visceral pain, including,
provides
pain;
pyelonephritis;
pain; central pain;
regional pain syndrome;
pain; acute musculoskeletal
injury/exercise
appendiGitis;
pain;
pain; acute visceral
cholecystitis;
'intestinal
hernias; etc; chest pain, including, cardiac Pain; pelvic pain, renal colic pain, acute
pain, including, labor pain; cesarean section pain; acute inflammatory, burn and trauma
pain; acute intermittent
pain, including,
endometriosis;
acute heIJles zoster pain; sickle cell
anemia; acute pancreatitis; breakthrough pain; orofacial pain, including,
pain; multiple
sclerosis (MS) pain; pain in depression;
-21-
leprosy
sinusitis pain, dental
pain; behcet's disease pain;
-'
PCTlUS2004/036297
WO 2005/042497
adiposis dolorosa; phlebitic pain; Guillain-Barre pain; painful legs and moving toes; Haglund
syndrome; erythromelalgia pain; Fabry's disease pain; bladder and urogenital disease, including,
urinary incontinence; hyperactivity bladder; painful bladder syndrome; interstitial cyctitis (Ie);
or prostatitis; arthritis, migrane, cluster headaches, trigeminal neuralgia, herpetic neuralgia,
general neuralgias, epilepsy or an epilepsy condition, a neurodegenerative disorder, a psychiatric
disorder such as anxiety and depression, myotonia, arrythmia, a movement disorder, a
neuroendocrine disorder, ataxia, multiple sclerosis, irritable bowel syndrome, or incontinence
comprising the step of administering to said patient an effective amount of a compound of
Formula I:
I
or a pharmaceutically acceptable salt thereof, wherein RA, Z, Y, r, and Ware as defined above
and in classes and subclasses as desribed herein.
[0029]
A preferred aspect of the present embodiment is where the disease, condition, or
disorder is acute, chronic, neuropathic, or inflammatory pain.
[0030]
Another embodiment of the present invention provides a method for treating or
lessening the severity' 6f. a disease~ condition: or disorder wherem the' disease, condition,' or '
.
.....
.
.
disorder is impllcated, in the, _activati~n,.of, voltage-:gateq . s~dium ,channels comprising ,
administering an effective amount of a compound of Formula I:
,',
I
or a pharmaceutically acceptable salt thereof, wherein R A, Z, Y, r, and W are as defined above
and in classes and subclasses as desribed herein.
[0031]
A preferred aspect of the present embodiment is where the disease, condition, or
disorder is acute, chronic, neuropathic, or inflammatory pain, epilepsy or an epilepsy condition, a
- 22-
WO 2005/042497
---
neurodegenerative
PCT/US2004/036297
disorder, a psychiatric
disorder such as anxiety and depression, myotonia,
arrythmia, a movement disorder, a neuroendocrine
disorder, ataxia, mUltiple sclerosis, irritable
bowel syndrome, or incontinence.
[0032J
A particularly
preferred
aspect of the present embodiment
is where the disease,
condition, or disorder is acute, chronic, neuropathic, or inflammatory pain.
[0033]
Another preferred aspect of the present embodiment is where the method comprises
an additional therapeutic agent.
[0034]
Yet another embodiment
NaVl.l,
NaVl.2,
of the present invention provides a method of inhibiting
NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaVl.?,
NaV1.8, NaV1.9, or CaV2.2
activity in:
(a) a patient; or
(b) a biological sample;
which method comprises administering to said patient, or contacting said biological sample with
a compound of Formula I:
..
:.
'.'
OJ;' a' pharmaceutically'
,
'
~..
..
.
~cceptable salt thet(mf, whe-rein
.
.'
'.
.
RA, Z~ Y, ,r,and.W
are as defined above '
and in classes and subclasses as desribed herein.
2. Compounds and Definitions.'
[0035]
Compounds of this invention include those described generally above, and are further
. illustrated by the classes, subclasses, and species disclosed herein.
definitions
elements
Handbook
chemistry
Sausalito:
shall apply unless otherwise indicated.
are identified in accordance
of Chemistry
and Physics,
are described in "Organic
As used herein, the following
For purposes of this invention, the chemical
with the Periodic Table of the Elements,
th
7S
Ed.
Chemistry",
1999, and "March's Advanced
Additionally,
- 23-
general principles
Thomas Sorrell, University
Organic Chemistry",
CAS version,
SUiEd., Ed.:
of organic
Science Books,
Smith, M.B. and
PCTffiS2004/036297
WO 2005/042497
......March,
J., John Wiley & Sons, New York:
incorporated
[0036]
2001, the entire contents of which are hereby
by reference.
The present invention provides compounds
B
monovalent,
such as RA, R
,
C
D
R and R
of Formula I with substituents
or divalent, such
;
that are
as A, B, and D. Those skilled in the
art will appreciate that for asymmetric divalent substituent groups, such as -Cl_6alkyl-NH-, and
-Ct-4alkyl-O-, there are two possible orientations relative to the parent structure.
As used within
the present specification, the orientation of a divalent substituent is set by its left/right orientation
relative Formula
appreciate
I as taught in the present specification.
Those skilled in the art will also
that this orientation convention is not relevant to symmetrical
divalent substituents,
such as -C(O)-, or -Ct-6alkyl-.
[0037]
-C(O)-,
For example for Formula I, where Z is N, W is present as la, RA is hydrogen, V is
Ring E is phenyl, RB is hydrogen;
-C3alkyl-NH-,
and A and Q are divalent
substituents
(A is
and Q is -Cta1kyl-O-), the following is the described compound.
o
O:>-'~·"-~~D
[0038]
-C(O)-,
For example for Formula I, where Z is N, W is present as la, RA is hydrogen, V is
Ring
E is phenyl, RB is hydrogen;
. ~Cialkyl~NIi-~d Q is -O-CiaIkyl-),'the'
'.'
.
and A and Q are divalent
substituents
(A is
following is the 'described compom.d .. :.,
.
0
.
())~~-~,~;~
V
H
[0039]
For example for Formula I, where Z is N, W is present as la, U is -CRr,
hydrogen, V is -CCO)-, Ring E is phenyl, RBis hydrogen; and A and
,
..
.
'.
. .
.'
(A is
and Q is -C]alkyl-O-),
- 24-
the following
RA is
Q are divalent
.
. substituents
.
is the described compound.
PCTlUS2004/036297
WO 20051042497
./
[0040]
As described herein, compounds of the invention may optionally be substituted with
one or more substituents,
classes,
subclasses,
such as are illustrated generally above, or as exemplified by particular
and species of the invention.
It will be appreciated
that the phrase
"optionally substituted" is used interchangeably with the phrase "substituted or unsubstituted."
general, the term "substituted",
replacement
In
whether preceded by the term "optionally" or not, refers to the
of hydrogen radicals in a given structure with the radical of a specified substituent.
Unless otherwise
indicated,
an· optionally substituted
group may have a substituent
at each
substitutable position of the group, and when more than one position in any given structure may
be substituted with more than one substituent selected from a specified group, the substituent
may be either the same or different at every position. Combinations
of substituents envisioned by
this invention are preferably those that result in the formation of stable or chemically feasible
compounds.
The term "stable", as used herein, refers to compounds that are not substantially
altered when subjected to conditions to allow for their production, detection, and preferably their
recovery, purification,
and use for one or more of the purposes
~m~odin'1ents;' a "~table compound
:0£
chemically
disclosed perein.
f~asible' com;~tJ.n«( is . otie . thai
In some
~s . not' ..: '.
sUb~antiaIiy altere~ when. k~p.t"at a temp~rllture .of· 40~C.p.r less, in. the. absence of .mois(irre or.
other chemically reactive conditions, for at least a week.
[0041]
The terms "aliphatic",
chain (Le., unbranched)
completely
saturated
hydrocarbon
or branched,
substituted
or un substituted
"cycloaliphatic"
hydrocarbon
or that contains one or more units of un saturation,
or bicyclic hydrocarbon that is completely
units of unsaturation,
molecule.
"aliphatic group" or "alkyl" as used herein, means a straight-
but which is not aromatic
or "cycloalkyl"),
chain that is
or a monocyclic
saturated or"that contains one or more
(also referred
to herein as "carbocycIe"
that has a single point of attachment
to the rest of the
Unless otherwise specified, aliphatic groups contain 1-20 aliphatic carbon atoms, Le.,
CI-20aIkyI. In some embodiments,
aliphatic groups contain
1-10 aliphatic carbon atoms, Le.,
In other embodiments,
aliphatic groups contain
1-8 aliphatic carbon atoms, Le.,
CI-lOaIkyI.
- 25-
..
WO 2005/042497
PCTfUS2004/036297
C1-salkyl. In still other embodiments,
aliphatic groups contain 1~6aliphatic carbon atoms, i.e.,
Cl-6a1kyl, and in yet other embodiments aliphatic groups contain 1-4 aliphatic carbon atoms, i.e.,
CHalkyl.
In some embodiments,
"cycloaliphatic"
(or "carbocycle"
monocyclic C3-Cg hydrocarbon or bicyclic Cg-CI2 hydrocarbon
or "cycloalkyl") refers to a
that is completely saturated or
that contains one or more units of unsaturation, but which is not aromatic, that has a single point
of attachment to the rest of the molecule wherein any individual ring in said bicyclic ring system
has 3-7 members.
substituted
Suitable aliphatic groups' include, but are not limited to, linear or branched,
or unsubstituted
(cycloalkyl)alkyl,
[0042]
alkyl, alkenyl,
and hybrids
thereof
such
as
(cycloalk:enyl)alkyl or (cycloaIkyl)alk.enyl.
The term "heteroaliphatic",
"heterocyclic"
aIkynyl groups
"heterocycle",
as used herein means non-aromatic,
"heterocyclyl",
monocyclic,
systems in which one or more ring members is an independently
embodiments, the "heterocycle",
"heterocyclyl",
"heterocycloaliphatic",
bicyclic,
or tricyclic ring
In some
selected heteroatom.
"heterocycloaliphatic",
or "heterocyclic"
has three to fourteen ring members in which one or more ring members
or
group
is a heteroatom
independently selected from oxygen, sulfur, nitrogen, or phosphorus, and each ring in the system
contains 3 to 7 ring members.
[0043]
The term "heteroatom"
or silicon (including,
means one or more of oxygen, sulfur, nitrogen, phosphorus,
any oxidized
form of nitrogen,
sulfur, phosphorus,
quatemized fonn of any basic nitrogen or; a substitutable
example
or silicon;
nitrogen of a heterocyclic
ring, for
N (at:. in 3,4-dmydr;2H~pyrrolyi);· NH· (as, in pyrroliciinYI) ·'or m+ (a~ in N~~~bstituted
.,. pY.r;0Ii~¥~)).'
.
.
[0044]
the
..
.
The term "unsaturated", as used herein, means that a moiety has one or more units of
unsaturation.
[0045]
The term "alkoxy",
or "thioalkyl",
as used herein, refers to an alkyl group, as
previously defined, attached to the principal carbon chain through an oxygen ("alkoxy") or sulfur
, ("thioalkyl")
atom, for example C14alkoxy refers to the alkoxyl group, methoxy,
ethyoxy,
propoxy, and butoxy, including for propoxy and butoxy, the straight and branched structures,
that is i-propoxy and n-propoxy; and n-butoxy, i-butoxy and
[0046]
The terms "haloaIkyl",
"haloalkenyl"
and
sec-butoxy.
"haloalkoxy"
means alkyl, alkenyl or
alkoxy, as the case may be, substituted with one or more halogen atoms.
means F, Cl, Br, or 1.
- 26-
The term "halogen"
PCTlUS2004/036297
WO 20051042497
[0047]
The term "aryl" used alone or as part of a larger moiety as in "aralkyl", "aralkoxy", or
"aryloxyalkyl", refers to monocyclic, bicyclic, and tricyclic ring systems having a total of five to
fourteen ring members, wherein at least one ring in the system is aromatic and wherein each ring
in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with
the term "aryl ring".
The term "aryl" also refers to heteroaryl ring systems as defined
hereinbelow.
[0048]
The term "heteroaryl", used alone or as part of a larger moiety as in "heteroaralkyl"
or "heteroarylalkoxy", refers to monocyclic, bicyclic, and tricyclic ring systems having a total of
five to fourteen ring members, wherein at least one ring in the system is aromatic, at least one
ring in the system contains one or more heteroatorns, and wherein each ring in the system
contains 3 to 7 ring members. The term "heteroaryl" may be used interchangeably with the term
"heteroaryl ring" or the term "heteroaromatic".
[0049]
An aryl (including aralkyl, aralkoxy, aryloxyalkyl and the like) or heteroaryl
(including heteroaralkyl and heteroarylalkoxy and the like) group may contain one or more
substituents. Suitable substituents on the unsaturated carbon atom of an aryl or heteroaryl group
are selected from halogen; -Ro; -ORO; -SRo; l,2-methylene-dioxy; 1,2-ethylenedioxy; phenyl
(Ph) optionally substituted with RO; -O(Ph) optionally substituted with RO; -(CH2)1-2(Ph),
optionally substituted with RO; -CH=CHcPh), optionally substituted with RO; -N02; -CN;
-N(RO)2;-NROC(O)RO;-NROCCO)N(R°h;-NRoC02Ro; -NRONROCCO)RO;
-NRONROCCO)N(R°h;
'.
"
.
.
.
-NRoNRoC02Ro;. ·-C(O)C(O)Ro~. -C(O)CHzCCO)RO; .:..COzRo; ~C(O)Ro;:' -C(O)N(R°h;'
"...
.
.' .
..'
.
. '.
.
.-OCCO)N(R°h; -S(O)2RO;-Sq2N(R°)z; .-S.(O)RO;.-NRoS02N(R~)2; -NRoS02R~;,·-CC=S)N(R°h;,
-C(=NH)-N(RO)2;or -(CH2)o-2NHC(O)ROwherein each independent occurrence of ROis selected
from hydrogen, optionally substituted Cl-6 aliphatic, an unsubstituted 5-6 membered heteroaryl
or heterocyclic ring, phenyl, -O(Ph), or -CH2(Ph), or, notwithstanding the definition above, two
independent occurrences of RO,on the same substituent or different substituents, taken together
with the atom(s)' to which each R ° group is' bound,
form
a
3-8- rnemb~red .cycloalkyl,
heterocyclyl, aryl, or heteroaryl ring having 0-3 heteroatorns independently selected from
nitrogen, oxygen, or sulfur. Optional substituents on the aliphatic group ofRo are selected from
NH2, NH(Cl-4aliphatic), N(Cl-4aliphatic)2, halogen, Cl-4aliphatic, OH, O(CI-4aliphatic), N02,
CN, C02H, CO2(CI-4aliphatic),O(haloCI-4 aliphatic), or haloC1•4aliphatic, wherein each of the
foregoing Cl_4aliphaticgroups ofRo is unsubstituted.
-27 -
wo
(0050]
peTIUS2004/036297
21)05/042497
An .aliphatic or heteroaliphatic
contain one or more substituents.
heteroaliphatic
heterocyclic
ring may
Suitable substituents on the saturated carbon of an aliphatic or
group, or of a non-aromatic heterocyclic ring are selected from those listed above
for the unsaturated
=0, =S, =NNHR",
carbon of an aryl or heteroaryl group and additionally include the following:
=NN(R*)z, =NNHC(O)R*,
where each RoOis independently
aliphatic.
group, or a non-aromatic
=NNHCOz(alkyl),
selected from hydrogen
=NNHSOz(alkyl),
or an optionally
or =NR*,
substituted
Cl-6
Optional substituents on the aliphatic group of R * are selected from NH2, NH(Cl_4
NOz, CN, C02H,
aliphatic), N(Cl-4 aliphatic)2, halogen, Cl-4 aliphatic, OH, O(Cl-4 aliphatic),
CO2(Cl-4 aliphatic), O(halo Cl-4 aliphatic), or halO(Cl-4 aliphatic), wherein each of the foregoing
Cl-4aliphatic groups of R· i~ unsubstituted.
[0051]
Optional substituents on the nitrogen of a non-aromatic heterocyclic ring are selected
from -R+, -N(Rlz,
-C(=S)N(Rlz,
-C(O)R+' -C02R+, -C(O)C(O)R+,
-C(=NH)-N(R+h,
-CCO)CH2C(O)R+, -SOzR+, -SOzN(R+h,
or -NR+SOzR+; wherein
R+ is hydrogen,
an optionally
substituted Cl-6 aliphatic, optionally substituted phenyl, optionally substituted -O(Ph), optionally
substituted
-CHz(Ph), optionally substituted -(CHZ)l-Z(Ph); optionally substituted -CH=CH(ph);
or an un substituted 5-6 membered heteroaryl or heterocyclic ring having one to four heteroatoms
independently
selected
from oxygen, nitrogen,
above, two independent
taken together
or sulfur, or, notwithstanding
occurrences of R+, on the same substituent
the definition
or different substituents,
with the atom(s) to which each R+ group is bound, form a 3-8-membered
'. .cycl<:>alkyl.;heterocyc1yl. li!1'l, or hete:rqaryl ring having .0-,3. ~e~eroat'?ms indepe~d~ntly, selected
,from -nitrogen~ oxygen. or
ot"R+ ~~ s'ele~ted fr~~
O(Cl-4 aliphatic),
sulfur:'
optional sobstituents op'the aliphatic group or'1he ph~nyl ring "
NHz, NH(CI~ ~iph~tic), N(C;'~'aiipha~c)z~'h~o~e~,'
NOz, CN, COzH, COZ(CI-4
aliphatic),
(:1_4 alipha~ic,'OH,
O(halo Cl-4 aliphatic).
or halo(Cl-4
aliphatic), wherein each ofthe foregoing Cl-4aliphatic groups ofR+ is unsubstituted.
[0052]
The term "Cl_2oalkylidene
chain" refers to a straight or branched
carbon chain of
twenty ca:b~n atoms, or less that roay be fq1ly ~a.tm:at~dor have oIJ,eor more units of unsaturati~n
and has two points of attachment to the rest of the molecule.
[0053]
Unless
otherwise
isomeric (e.g., enantiomeric.
structure; for example,
stated, structures
diastereomeric,
depicted
and geometric
the R and S configurations
double bond isomers, and (2) and (E) conformational
- 28-
herein are also meant to include
(or conformational)
for each asymmetric
isomers,
all
forms of the
center, (2) and (E)
Therefore, single stereochemical
PCTlUS2004/036297
WO 2005/042497
~
isomers as well as enantiomeric,
the present compounds
diastereomeric,
and geometric (or conformational)
mixtures of
Unless otherwise stated, all
are within the scope of the invention.
tautomeric forms of the compounds of the invention are within the scope of the invention.
For
example, for compounds of formula I:
wherein Z is Nor 0, one of ordinary skill would recognize that a suitable tautomer is as depicted
above.
When the Z group of formula I is CR, one of ordinary skill would recognize that a
suitable tautomer is as depicted below:
y
w
[0054)
Additionally,
unless otherwise stated, structures depicted herein are also meant to
include compounds that differ only in the presence of one or more isotopically enriched atoms.
For example, compounds having the present structures except for the replacement of hydrogen
by deuterium or tritium, or the replacement
of a carbon by a 13C_ or 14C-enriched carbon are .
withiri the scope 'of this invention~' Such compounds
or pr~be~ in:biolo~cai
assays:.
......
....
are useful!
for example, as analytical t061s
-
3. Description of Exemplary Compounds:
[0055]
In certain embodiments,
the present invention provides
a compound of formula I
wherein V is -8(0)2-.
[0056]
. In other embodiments,
the present invention
provides
a compound
of formula 'I
wherein V is -C(O)-.
[0057]
In one embodiment,
T is a
Cl-6
straight
or branched
methylene unit of T is optionally replaced by a C3-6 cycloaliphatic
aliphatic chain wherein
group.
[0058]
Also preferred is a compound of formula I wherein Ring E is phenyl.
[0059]
Also preferred is a compound of formula I wherein Ring E is naphthyl.
- 29-
a
WO 2005/042497
PCTlUS2004/036297
[0060J
Also preferred is a compound of formula I wherein Ring E is pyridinyl.
[0061J
Also preferred is a compound of formula I wherein Ring E is thienyl.
[0062]
Also preferred is a compound of formula I wherein Ring E is furanyl.
[0063J
Also preferred is a compound of formula I wherein Ring E is quinolinyl.
[0064J
Also preferred is a compound of formula I wherein Ring E is benzofuranyl.
[0065]
Also preferred is a compound of formula I wherein Ring E is 3,4-dihydro-2H-
chromene.
[0066]
Also preferred is a compound of formula I wherein Ring E is
2,3-dihydrobenzo[b] [1,4]dioxine.
[0067]
According to one aspect, the present invention provides a compound of formula I,
wherein Ring E is a preferred group as described above and said grOup is in combination with
the remaining variables of formula I as set forth in the classes and subclasses described herein.
[0068J
In certain embodiments, each RA of fonnula I, when present, is independently R6,
OR6, CN, or halo. In other embodiments, each RB of formula I, when present. is independently
OR6• N(R6)2' NR6C(O)R6• halo, R6, C(O)R6, or N02.
(0069]
In other embodiments, the A moiety of formula I, when present, is -T_NR5_, wherein
T is a Cl-6 straight or branched aliphatic chain. Such A moieties include -CH2CH2N(CH3)-,
-CHzCHzNH-, -CHzNH-, and -CH2CH(CH3)NH-.
[0070]
In certain embodiments, the present invention provides a compound of formula I
wherein A is·.,-T-NH- where~ Tis a,C{6 straight·or branched aliphatic chain wherein a· ..
,
methylene"ucit
of T is repla~d. by a C3.<;<.;yclo8.Iiphaticgroup.
S'uchcycloaliphatic
groups..
.
.
'.
.,
-.'."
'.'
'.
','
,
include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl groups.
[0071]
In other embodiments, the Q moiety of formula I is a Cl-4 alkylidene chain wherein
one methylene unit of Q is replaced by -0-, -NH-, or -S-. Such Q moieties of formula I include
-CHzCH20-. -CHzO-, -OCHz-, -OCHzCHz-, -CH(CH3)0-, -NHCHz-, -C(CH3hO-, and -CHzS-.
,[0072]. ,
As .described above, for compounds of the invention of Formula I, Z is 0,. N or C.
Accordingly. in certain embodiments, where Z is N, the corresponding compounds have the
structure of Formula II:
- 30-
PCTlUS2004/036297
WO,;f',OS/042497
IT
[0013] .
In certain embodiments of Formula II, Y is hydrogen, Ring E ,is phenyl, and W is
formula la, wherein V is -CCO)-, as shown in Formula ITa:
- 31 -
PCT/US2004/036297
WO 2005/042497
....
CN, CI-6 aliphatic, -N(R6)z, or halogen.
Such RB groups include methyl, ethyl, butyl, isopropyl,
chIaro, fluoro, bromo, N(Me)z, CF3 and -CHzphenyl.
According to another embodiment,
the
present invention provides a compound of formula Ua wherein the benzo ring is substituted at
one of, or both of, the C-4- and C-5 positions with tert-butyl, fiUOTO,or methyl.
[0083]
fonnula
According to another embodiment,
IIa, wherein A is -CH2CH2NH,
or -CH(CH3)NH-,
6'
independently
CI-6
the present invention provides a compound of
,
.
6
aliphatic, -N(R )z, -C(O)R
,
Q is -CHzO-
.
or halogen.
B
Such R
B
and each R
.
is
'
groups include methyl,
chloro, bromo, ethyl, N(Meh, -C=CH and C(O)CH3•
[0084]
Yet another embodiment of the present invention relates to a compound of formula
Q is -CHzO-, -NHCHz-, or -CH(CH3)O-,
ITa, wherein A is -CHzCHzNH-,
. Of h~Qgen ..
.,:
..
indepep.dept~y C1:6 aliph!l;tic:.. ~9R6,
.
chIoro, bromo; and flu6ro.
[0085]
Accordirig to' still ~other embodi~;mt,
B
Such R
.
-CH(CH3)O-,
or -C(CH3)20-,
and each R
is
gt:pups, ~ncll,1,demethyl, et:9yl,..:OMe"
the"present'inventio~
is independently
provides a compound '~f
Q is -CHzO-, -NHCHz-, -NH-,
formula Ha, wherein A is -CHzCHzNH- or -CHzCH(CH3)NH-.
B
B
and each R
6
CI-6 aliphatic, _OR , or halogen.
Such RB groups include methyl, ethyl, -OMe, chloro, bromo, and fiuoro.
According to another
A
embodiment, the present invention provides a compound of formula Ua wherein r is 2, each R
is fiuoro,and is present at the C-4- and C-5 positions.
[0086]
In certain embodiments, the present invention provides a compound of formula Ha,
wherein A is -CHzCHzNH-, Q is -CHzO-, and each RB is independently
halogen.
In other embodiments,
wherein A is -CH2CH2NH-,
bromo, or chIaro.
the present invention provides a compound of formula IIa,
Q is -CHzO-, and each RB is inependently methyl, isopropyl, fiuoro,
In still other embodiments,
the present invention provides a compound of
B
fonnula Ha, wherein A is -CH2CHzNH-, Q is -CHzO-, and each R
fluoro. or chloro.
is inependently
methyl,
In yet other embodiments, the present invention provides a compound of
B
formula IIa, wherein A is -CH2CH2NH-, Q is -CHzO-, and each R
is inependently
bromo, or chi oro.
[0087]
CI-6 aliphatic or
Also preferred is • compound of Formui.
- 32-
na. wherein
A is
~,A
methyl,
PCTlUS2004/036297
WO 2005/042497
~
I
l{'-~
[0088)
[0089].
Also preferred is a compound of Formula ITa, wherein A is
. Also preferred is a compound
of Formula
a
IIa, wherein A is
wherein U is -CHr or -CHzCHr.
[0090]
In certain embodiments of Formula II, Y is hydrogen, Ring E is phenyl, W is
Formula Ib and D is a bond, as shown below as fonnula Db .
.....
.
,. ,.
fib
or a pharmaceutically
acceptable salt thereof, wherein RA, r, RD, and s are as defined above and
herein.
[0091J
Preferred is a compound of Formula lIb, wherein RA is R6 or halo.
In certain
embodiments, RA is methyl, chIaro or bromo.
[0092]
Particularly preferred is a compound of Fonnula lIb, wherein RA is methyl.
[0093]
Also preferred is a compound of Formula lIb wherein RD is halo, OR6, N(R6
n,
NR6C(O)R6, or two RD are taken together to form a methylenedioxy
or ethylenedioxy
group. In
D
certain embodiments, R is -oH, -N(Et)z, -OMe, -NHC(O)CH3, fluoro, or chloro.
[0094]
According to another embodiment,
the present invention provides a compound
formula ill:
ill
- 33-
of
PCT/US2004/036297
WO 2005/042497
or a pharmaceutically
B
acceptable salt thereof, wherein A, Q, RA, r, R
,
and s are as defined above
and herein.
(0095]
In certain embodiments, each RA of formula nI, when present, is independently
R6,
OR6, CN, or halo, In other embodiments, each RB of formula In, when present, is independently
OR6, N(R6)z, NR6CCO)R6, halo, R6, C(O)R6, or NOz,
(0096]
In other embodiments, the
Q moiety of formula III is a Cl-4 alkylidene chain wherein
one methylene unit 0(0' h replaced by
-0-, -NH-, or -8-,
Such Q moieties of form~ia
nr
include -CHzCHzO-, -CHzO-, -OCHz-, -OCHzCHz-, -CH(CH3)O-, -NHCHz-, -CCCH3)20-, and
-CHzS-:.
(0097]
In'other embodiments, the A moiety of formula In, when present, is _T_NR6_,
wherein
T '"is a ...
Cl-6 "..
straight
or branched
aliphatic
chain.
Such A
moieties
'
. .
.." ...
'.,"
.,..
..
.,. . include.
,-'."
.....
"
'",
,
-CHiCH2N(CH3)-, ~CH2CH2NH-, -CH2NH-,and
-CHiCH(CH3)NH-;
.
. , [0098]'
III certain
embodiments. the presentinv'enti~n
. ..
pr~vides a ~omp~und offormul~
m'
wherein A is -T-NH- wherein T is a Cl-6 straight or branched aliphatic chain wherein a
methylene unit ofT is replaced by a C3-6 cycloaliphatic
group. Such cycloaliphatic groups
include cyc]obutyl, cyclopentyl, and cyclohexyl groups.
(0099]
According
to another embodiment,
formularY:
the present invention provides a compound
of
~~
>-C(/v,J0:tR'
o:
#
(RA>r-I-
.--PN
o
IV
or a pharmaceutically
acceptable salt thereof, wherein V. Q, RA, r, RB, n, and s are as defined
above and herein.
[00100]
ill certain embodiments, each RA of formula IV, when present, is independently R6,
OR6, eN, or halo. In other embodiments, each RB of formula IV, when present, is independently
OR6, N(R6)2, NR6C(O)R6, halo, R6, CCO)R6, or NOz.
(00101]
In other embodiments, the Ring E group of formula IV is phenyl or a 5-6 membered
heteroaryl ring having 1-4 heteroatoms independently
·34·
selected from nitrogen, oxygen, or sulfur,
PCTlUS20041036297
WO 20051042497
'-4
or a 3-7 membered monocyclic heterocyclyl ring having 1-4 heteroatoms independently selected
from nitrogen, oxygen, or sulfur,
Such Ring E groups of formula IV include pyridyl, thienyl,
furyl, and pyrazolyl.
[00102]
In still other embodiments,
bicyclic
aryl ring or an 8-10 membered
independently
.
the Ring E group of fonnula IV is
bicyclic heteroaryl
ring having 1-4 heteroatoms
selected from nitrogen, oxygen, or sulfur.
",
'.
.'
include naphthyl, quinolinyl, 3,4-dihydro-2H-chromene,
[00103]
an 8-10 membered
Such Ring E groups of formula IV
. .
.. .
and 2,3-dihydrobenzo[b](I,4]dioxine,
In other embodiments, the Q moiety of formula
one methylene unit of
IV is a Cl-4 alkylidene chain wherein
Q is replaced by -0-, -NH-, or -S-. Such Q moieties of formula IV include
-CH2CH20~, -CH20-, -OCH2-, -OCH2CHr, -CH(CH3)O-, -NHCH2-, -C(CH3)20-, and -CH2S-,
[9~1,~4J.. Acc?rding to a~othe~ .~p«c~ of ~e presen~.i.~ventiont .qne of V
According to yet' another aspect of the presen.t irivention, both of V and Q
~4 Q
are Iibond.
[00105]' "Rep~sentativ~ 'coz:npo~n"dsof formula I 'are 'set fo~ in Tabie 2 belo~.
·35·
is. a. qon4.
PCTlUS2004/036297
WQJOOS/042497
"
T
<J
T"~O
2.
t-V
"C
~
~.
Q
u
"C.
o.
. ..
N
~'lIo
U
I
\0
'",
T
0
...
P
d
1
~
Q
t
0
en
Q
U
=
::l
~
~
B'
u
u
~
i3en
Q)
"Cl
~ ~~
u
Q)
«l
.....
PCTlUS20041036297
WO 2005/042497
'+
Y-t'
'C
g
.-S'
-,'
0
U
.-
({
b
~~
%':r'
-.
"'='
S'*
-<t
U
I
I
1eo
u
Q
Oy;:"
0
'C
e-*
u
""'
PCTIUS20041036297
WO 2005/042497
r
"~I 9
't:l
0
:r:r
§
0
.~(
S-o
u
b
..
!C
'r::.
\D
~'ll:
I
00
en
I
u:'
.. ~
~, .',6..
o~
¢
o·
't:l
.%~
i~
~
S-o
U
.
'"
..
..
0
" Ii
't:l
S''ll:
U
In
WO 2005/042497
peT IUS2004/036297
'T"
"-;
~
't:l
=
::l
0
S-
.
"'''
0
u
'
b
.
,
'i~
"
0-
U
:
Mr
~I,4,
'0'
~ ..
't:l
~
~
Q
U
-
:Z'
J~.
0
a-~
't:l
u
I"'-
PCTlUS2004/036297
W0-.f.005/042497
Q
~F
't:l
~
~
u
o
I
~t
0.....
~.... .,,0
,.. '!.
T
..
. "::lft:jj:
:::
u
,
o
"",
1s-
o
U
Q
),1"
'!.
"%
't:l
S':jj:
u
!::!
PCT/US200~/036297
WO 2005/042497
"p
~,..
't:l
l=l
g
:r:r
a-
Q
O~
Co)
0
'"
U
",
..
..
'tl
:!l
S"=I!:
C,)
.....
.
"'"
I
",,\O'Q
'tl
§
Q
~
u
:t',.. .....'\,
.i.
.
(I
b
'tl
~~
C,)
;!;
PCT/US200M036297
WO 2005/042497
'l:l
~
t
0
U
-P
/'"
:z-r
o.=)
a.
~
"
<.J
(]
...
,
...
,
i*
....
t--
U
.
~,
I
.~
i
g,
~
u
:1
. :zo.,%,
o=) ~
cf"
"tl
S'*
U
10
.....
PCTlUS2004/036297
WO 2005/042497
Q
l~
'g
::I
0
~
0
u
°6'
...
'C.
S':ll:
0
N
U
I
.
~
,,.
;
Q,...-u'"
:
::
'C
§
0
~
0
u
'C
S':tl:
U
.L
b
0\
PCTlUS20041036297
WO 2005/042491
.;'
<.J
,&
~ lyO
]
t.
T;!!
a0
u
Z' .
0
'
. !C ..
S'=II:
.'
..
N
C'I
U
.
~
I
q
~
'C
§
c
t
Q
U
9'
'-I.
-;s:
'C
t=ll:
U
....
C'l
..
peT IUS2004/036297
WO 2005/042497
~
:7
~ I
]
0
~
-:r%'
.i.
0
t
0
U
.
,
0
"
,
't:I.
Ct.' .
.
'<t
N
~:jj:
.
.
~
i'O
7
<J
'
~,
yo
0
't:I
~
a-
0
u
%'7
J.
b
't:I
S"'!l:
U
1::)
peTIUS2004/036297
WO 2005/042497
'1
Q-:~
'\..
"C
~
'S'
0
U
~o
'
..
.
.
..
"C
S'*
U
..
'"
N
I
..,.
\0
I
JOY'
u
.
"C
§
0
a'
Q
U
1b
"C
S''Il:
U
1/1
N
PCTfUS2004/036297
WO 20051042497
Q;.
'1
't!
§
~.
,,,,.
~
U
Q
b
.
1*
a-
N
U
,
~,
q
~r
'0
§
t
Q
u
9'
<:i
'0
t~
U
co
C'l
-,
PCTfUS2004/036297
WO 20051042497
T
'Q
.-t:J
'C
1:1
::l
0
S'
..
0
_
0
U
V
. .
"CI.
'"'"
~=Il:
U
,
~,
~",
't:l
g
a-
0
U
P
"''''
o=)
0
P
V
'tl
S'=Il:
U
0
<'l
T
PCTlUS2004/036297
WO 2005/042497
I
r
..
y
Op
"t:l
g
S-
Q
U
~%
%'"
,
0
\
<J..
or
..
i~
r--
<'""l
U
,
.
0\
'<t
g"
"t:l
~
a<:l
U
OJ-p'
<J..
or
"t:l
a-~
U
'"
<'""l
PCTlUS2004/036297
WO 2005/042497
Q
1S'
Q
U
"!'
~~(>
"b"
%
"
'C-
I'll:
......
'<:t
U
I
......
trl
I
-
1
S'
Q
U
Q
.~~
~ ;}
"Cl
S''*
U
'D
'<:t
peT IUS2004/036297
WO 2005/042497
7~
U
0"
'tl
g
S'
e
U
¢
~ ,(0,
".1
0
b
"
,
.'C
~
8'=11:
U
,
~,
'C
§
8'
c
u
Q
~ ,(0
0
.Z"
".1
b
'd
S'=II:
U
....co
WO 20051042497
peT ruS2004/036297
o~ ","
'"
¢
"',J:.0
'tl
§
Q
.S'
:Jr.
0
,,{
Q
U
0
'C
S'*'
C'l
Irl
U
.
t<l
Irl
I
'C
6
Q
t
0:'".;~~
.
1.&.
.
~
.,.
8
"CI
S''Il:
U
....
Irl
PCTfUS2004/036297
WO 2005/042497
p
;;:
-
.
'tl
§
,..,..
0
.~
O~·
0
c.>
b
..
.
..
.
"'C.
g..
S'll:
c.>
.,.,
~
,
:1i,
I
'C
~
S'
Yr.~
:z:-.:
r
0
u
U~
T'
'tl
6'*
c.>
0
"<t
WO 2005/042497
PCTlUS2004/036297
¢rt
oe
~
~
e
U
TOT;::
./(
U
..
"Cl
8'=11:
""
on
U
,
~,
.Yt~
I ".''
"~,
9
...
.";'::;;'
0
"Cl
g
S'
Q
t.>
%,1:
~.
0
~=
..
"Cl
8''ll;
t.>
<"l
on
0
'(,I
WO 2005/042497
PCTlUS2004/036297
~
9-,
7.~J::
"0
g
S'
...
1
Q
u
0
-
't:l
Q"
.
.,
El'll:
\0
\f)
U
,
\f)
\f)
,
v
~
1:0.
"0
~
l:l..
u~
T"::i
.
..
i
b
't:l
S''ll:
U
:R
(I
'-I.
<r
PCT/US2004/036297
WO 20051042497
-'
.
""'.
10
,....,
u
,..'"
,
~.UXO
'l;1
g
S-o
O
t'..)
';'1%
i
"'<:",
U
'l;1
!#
¢
t'-
""'
PCT/US2004/036297
WO 2005/042497
...,
'2:
e-Q
"0
..
§
a-
..Q
z""
,.'7
0
u
..
"
"0
. S'=II:
6
\0
U
,...,
.,.,
,
..
.'YY
.%
' r
~
0
;.,.1:
iS'
0
U
T,:I:'
i·
~z
0
"0
5=11:
.,.,
0\
::{'
.
~
'"
0
.,.
WO 2005/042497
'--r
'tl
§
0
l:l<
S··
PCTlUS2004/036297
..
..
~
-:r':r
~.:z'.
Q
Q
U
U
(j
'[-
-e·'lt:
..
.~
u
I
co
'",
'tl
~
~
u
Q
.... ~
~
"%,%
-
~:r
Q
u
"Cl
t'lt:
U
..-<
10
..
-
WO 2005/042497
PCTfUS2004f036297
-r
~
':<
0
.
~
~
"Cl
§
0
S-o
C,)
'"
~.%J::
,.1.
b
..
'Cl
~.
f=ll:
C,)
,
'",
lr1
J?
,=0,,(0
0....1.
h-
II
'Cl
g
a-
0
C,)
..
1
0
b
"Cl
S'=lI:
C,)
'"
IC
WO 20051042497
PCTfUS2004/036297
.......
~
I
b
~
I
b
'tl
§
T~.A:
Q
a-
8
7"~
Q
"
i*
u
\l)
\l)
.
@
I
'H
~J:0
t
T
'tl
g
o:r"Z'_.
a-
0
u
or.::,
~Z'
b
'tl
a-*
u
\a
.
.
·0
PCTlUS2004/036297
WO 1005/042497
"\
.. ..
i'Q
'%
U
'%
u
't:l
~
'%'2J::.
.S'c
..t
u
b
..
..
. ..
..
'l:l
.S'*
U
00
'D
,
\0
I
P"'~2J::
i
.....
u
't:l
~
S'
0
U
'7':1"
~=
U
"0
S'*
U
l'\0
~.
..
u
.
"'.
PCTlUS2004/036297
WO 2005/042497
'""
0
I
~O
oy
o
"I:l
§
0
~
0
,
I.d-
i~-'"
.
U
7_%
~Z'
0
.
,
!CI
Q,'
8=11=
0
t"'
U
,
~,
..
f(C~
'"
~...
'"
"I:l
~
~
0
u
TJ:¢
...2 ·
0
"I:l
~=II=
U
.
01
\Q
~
":"
PCTfUS20041036297
WO 2005/042497
oy
JY~
(l
1
;e"'o:
~
U
~':Z
~:r
0
..
..
..
o.
'tl
'S'*
U
M
I"-
I
~,
0.:0
o
°0~~
iS'
Q
U
"":or
i"':
"':,.: or
0
'l:l
8*
.
....
I"-
..
wo
2005/042497
PCTfUS2004/036297
.y6J
"d
g
i~'~
~
0
U
OZ.Z'
~%
0
,',
,
'
"d'
t*
U
"
:
"
.,
"""
r--
, ,
,
'd'
\0
,
oy
<l
"Cl
g
i~'T
t
0
U
JY-'
:,,%
~:
0
"d
~=II:
U
r-'"
.
..
WO 2005/042497
PCTlUS2004/036297
~
"
~U
oy
o . Q
'l:I
g
1
7
~
8
-;t'tZ
'"
~-;zr
•
0
'"
"d
S'=It:
'Q
I"-
U
,
:a.
'l:I
o~
§
"~
i"o
Q
.
~
U
J9
,
;.)Z
"
~Z'
0
"d
S"=It:
U
\r)
r--
WO 2005/042497
-.
PCTlUS2004/036297
C£rC
oy
't:l
g
'S'=>
i%'
"
u
.'
':r
~
Q
•
'8
..
"C
S':lI:
'.
DO
l-
e.;
I
\0
\0
I
9Y.J?
"C
g
!
.
i"';%
'Z't:z
~%
b
't:l
S':lI:
U
r--
r-
o.%
,
PCTfUS2004/036297
WO 2005/042497
~~
oy J?o~
"Cl
§
~
" X~·~
0
S0
U
'%'z
".
~:::!
0
..
"
~..
"
"Cl
I'll=
U
:
"
, "
0
00
,
lO,
.~
oy
0
"Cl
§
0
i%'
I
8
·%'2
", .. '"
0
"Cl
S'=#;
U
0.
l'-
;;:...,
.
peTIUS2004/03629
WO 2005/042497
7
'~
,VU
1,,7
oy
0
"CJ
=
l~·
::I
0
'S'
"
0
'%~%
U
% '
~%
0
"CJ
S'~.
,
,
C'l
00
U
,
00
\0
,
0'0
.
.~
°r~~
0
'l:l
~
i%O%
S'
Cl
U
T ..Z'
"::z
0
"CJ
~=II:
u
l·
....
00
PCTlUS2004/036297
WO 2005/042497
-.t'
~:X)
oy
b
I
"Cl
§
0
S'
"T'2{'~'
0
U
U
..
..
'8.. .
a'll:
..,.
00
u
,
$
1
Q
~;%)%
%.
:r.
"Cl
§
~o
oyO'°
0
S'
0
U
i%>T
T.",
~"
0
"Cl
S''ll:
U
<"l
00
-.
PCTlUS2004/036297
WO 2005/042497
-,'
8
ct
f ~
]
0
0
t
' ,
~o,
,:,.:r"
0
U
z~
17'
~I
,
'
..
-=t* .
,
,
\0
00
U
.
g
,
0.,)"JY"
iZ'~
"C:l
IJ~
,
~
a-0
U
7.Z'
tr ,
~:If
0
't:l
S'*
U
In
00
peTIUS2004/036297
WO 2005/042497
---r
u
/>
0Y\O
't:l
=~
S' ..
Q
Q
U
..
"
~-%~:'::":
0
..
't:l
~~
00
00
PCTlUS2004/036297
...
WO 2005/042497
oyoJCX:
u
"CI
=:c:
--X%-:
~
C
U
":::z:
%'='
0
-.'
"
,
~
S'=II:
.'
0
Q'\
U
,
~,
.,JJI'
o
§
0
s-
o
U
<:i
1%0%
..
'%'z.
~:r
0
"CI
~=II:
t;,)
Q'\
00
PCTlUS2004/036297
WO 2005/042497
4r
...
1
"*"
.~
"4"
i~>T'
U
7' ~2'
.'
'::::z
b
~
. .
. 'tl
S'~
U
:
'.
N
Q\
I
~,
;oro
. ::7'1 . '
.~.':
00
'tl
~
!
.....,.0
i%'T
,
'%'%'
~='
b
'Cl
S':jj,
u
....
'"
wo
PCT/US2004/036297
2005/042497
~
xrr~
oy
<i
°
"'='
~
~.
°
"I..
~
i~'ir
·t
Cl
U
'%...2'
~:z
b
..
.,
';I
t'"'
..t-
'"
U
.
~
r,
..f'.~
'10'
Oy\°
'l:t
g
l~·~·~
t
Cl
U
'Zl:z:·
~.:r
U
';I
B':tl:
U
... <:1
~
'"
'"
w~
PCTlUS2004/036297
2005/042497
...
."
U
0"
,~
"d
~
aQ
U
:.:X:~:
<1, .
. u...
:r
U
"d
S'*
\D
'"
U
,
~,
,2
-=
"l:l
g
i
t~
u
0
~o
.
i%':r
.
T,,;,'
.~
....
:r
~".
Q
"l:l
lr\
~=lI:
'"
Q
WO 20051042497
PCTIUS2004/036297
4
~
:r
)~
oy ~
'l:l
§
. i"",r,
Q
.mQ
C.l
:'%
~2
•
0
".
."d
S'*
C.l
,0' •
.'
'.
Cl\
""
,
ID
t-I
~
~
7
oyL1l'~~
~,~l::'~
0
~
'l:l
g
S'
Q
C.l
"d
S''II:
C.l
t-Cl\
WO 2005/042497
PCT/US2004/036297
I
~-P
§
:X:'%
0
S-o
8
u
.f_~
.
"tl
,"
0
8
~=Il:
u
,
rrI
~
%.
<J.
;f
I .
O.,J ~
i%'
"0
==
0
aQ
U
%~%
~
..
T
U
"l:l
t=ll:
U
'"'"
&'~
PCTlUS2004/036297
WO 2005/042497
o~
't:l
1.4- .
°0~
1%0
§
0
S'
7'
0
U
7':::r
~%
U
..
't:l
....~
S'=ll:
U
,
.
00
t"-
~n
.
o
't:l
g
S'
0
U
°0~
1%"%
%,%
~%
U
't:l
S'=ll:
u
I
,...,
0
,...,
V
PCT/US20041036297
WO 2005/042497
:xr~
~I
Oyl~
'l~':
't:I
g
.~
'.
c
U
:.",
~%
U
'
.
.
.
. 'C
S'~
....
·0
.....
C,)
,
0\
f-o
,
'C
§
Q
t
Q
C,)
8~i=
<J.
•
:)
&..,
.x-... .
'C
<'1
~~
0
'%'.2'.
0
....
PCT/uS2004/036297
WO 2005/042497
'l:l
S=
'S'
0
u
2'
£-0
"i"'''~,
..
..
'C
\0.
0
S'*
.....
U
,
o
co
,
1S'
0
u
'd
S'*
U
. I
..
.~
~
~,%.~%
£-0
1f'l
0
'
PCTlUS2004/036297
WO 20051042497
~
'tl
~
t
Q
u
r~
O~--o
"'....,.~
'Cl
...
:
cO
S'*'
U
0
.....
I
,....,
00
I
'tl
i
t
0
u
'tl
t*
U
s:
%
I'
.. :
:'~
',' z
5~
.....
....
0
Ci
WO 2005/042-197
, PCTlUS2004/036297
~
...<J~
I
~P.o.
a··
Q
u
Z'
1#
'':1'.;'
Z
5-D
'C
0
.....
S'*
u
....
.
&l,
'C
~
S'
Q
U
'C
S''lI:
u.
r
.-{)
·~.I
'%~2.
~::!'
cr,
~
0';"
PCTlUS2004/036297
WO 2005/042497
.J'
0':'-
'tl
§
0
B'
u0
.,2
~--o
i
, 'tl .
~
......
B'=lI:
U
I
t""l
00
I
'tl
~
a0
U
'tl
~=lI:
2
CO
.Q
T.",
~'"
......
PCTlUS2004/036297
WO 2005/042497
)-
it·
Q
u
:.2'
£-0
<J
.&- <J
~~;i-.
;:r'
.,
..
"C
..
SO'#:
U
'<to
....
....
.
I
.
;;:t;
"C
~
~
Q
U
"C
S''#:
U
(:2'J
o
'0
e-o
''%12'
.. ~Z'.
........""'
.
WO 2005/042497
PCTIUS2004/036297
)-
0
D
Oy\
"Cl
§
Q
g.
~
U
i~'
'I-,
..
T
T'Z
~%
T
0
,
"
"Cl
~*
\0
.-<
.-<
U
,
\f')
00
,
'"........
PCTIUS2004/036297
WO 2005/042497
~
~
T
U
~
1
o~
:
i%;r
.~
Q
U
'%':z
~:r
b
...
.,
;
:
"Cl'
t:jj:
..
.,
.,
00
.....
.....
u
.
\<:l
00
,
I
I
;po
£(0
o~
"Cl
~
l:l<
Z
~
u
i
.
-:r,z
'
"::z ,..
b
"Cl
t:jj:
U
t--
.....
.....
.~
PCTlUS2004/036297
...
WO 2005/042497
,J?
oy
D
"C:l
§
i~-~
0
t·
Q
U
712
~::'
0
..
'0
t'll:
U
.
..
.
0
N
"'"'
.
.
t"-
oe
yM~
'0'
"C:l
g
i"'~';'
aCO
U
. ..
-:r.Z'
~;r
0
"Cl
S'll:
U
0-
"'"'
rl
~
.,.
PCTIUS2004/036297
WO 2005(042497
ii,
I~
'tl
§
O~
i%' .
0
.~
0
U
:<'%
'>::% .,.-
0
..
..
'tl
'S'*
M
M
.....
U
I
""
00
oyu.0
1S'
1%·
0
U
:<,=
~%:<
0
'l:l
S"""
U
.....
M
.....
PCT/US2004/036297
WO 2005/042497
~
I
.
a..
00
'C
~
~
u
Q,JO¥
.£~~~
'%'''. ~,.
b
'C
t=ll:
U
<"I
C'l
-<
ij
WO,.2005/042497
PCT/US20041036297
,
o
'",
/1
s'<9.
'tl
-r':i!:
~
.~
~
Co>
SO"
. !J
0
'tl
~=ll=
Co>
\0
N
:
-
WO 2005/042497
PCTlUS2004/036297
~
",,0
O~
'"
.J)
Oyl
i~.
",.,..
",in
Q
U
C
.,
s~
"CI
.
.
."
'.
...
"
"
0'
N
,
....
0\
,
00
N
....
PCTIUS2004/036297
WO 2005/042497
oy¥~ ~
~I
'l:l
~
=
0
%:r
'9'
%f~:r
0
C,)
b
"
'l:l
S'=n:
'
......
"
.".
<')
......
U
.
.
~
r-O
'l:l
~
0
S'
0
C,,)
oy
~o
"''''
",j",:r
0
'l:l
S'=n:
c.>
...,
0
....
I
:
PCTlUS20041036297
WO 2005/042497
~
9
!~
~
"Cl
0
~
~
c:>
..
'",..
"
'?
u
0
'I ~
u
.,
.'
"d
os'**'
M
M
......
U
o
M
0\
o
~
.':r'
JQt'
,cy ~
I
"Cl
§
Q
!
.~
.
'%
~Z.':l"
1
""~ ."',..
0
"Cl
8'**'
U
,if.
N
.....
M
peT IUS2004/036297
WO 2005/042497
...u~ ...u~
l)
%
-=B
0
e-
"
~~
.,.". "'::".
0
U
t--U
"
"
.~
-.
-==.'
....
....
'\0
8=11:
U
,
g,
,
I
u
O='!.".
p
PCT/uS20041036297
WO 2005/042497
r
,;
<J
'l:l
g
~.
Q
U
~i
0 ....
G~-b
T%
"'-'::~
..
'
1:*
U
.
00
••
.....
'"
,
."
'",
U
?
. -0
I
1
~
8
.~
l
'l:l
s*
U
r--
....
<"l
WO 20051042497
peT IUS2004/036297
r
X
"Cl
§
0
S'.
P
0
u
u
..
"Cl
~
....
~'*"
u
I
\0
0-.
I
.
,;;,
:r
"Cl
"~
~
a8
"',..
.
<:J...
,.,.
'~:I"
LO
"Cl
S'=ll:
U
'"....
c<1
WO 2005/042497
peT IUS2004/036297
r-
~"
'l:l
=
::t
Q
t
'!'~
,"''''
P
Q
u
<l
'
,
"
'Cl
..
S'*
U
'
'
•II!>
v
....
,
t"-
o.
,
£
I '
'l:l
g
~
8
_' , b._
<J
' . .
p
z '9". :z:>".
ij
'Cl
S'*
U
;<1
'"
v
..,.
WO 20051042497
PCTfUS2004/036297
,.....
u
....
0
~ I
'C
§
"5
0'"
0
g.
So
o
0
~
0 - ",,'7
0,.,,-.
.
P
U
u
,"
..
00
'C
t°:il:
U
00
Co
-.to
.0
0-
......
1
00
'"
1
:9'1 .
'"Cl
o~
~
a0
u
~
'Z'7
P
u
'C
~'ll:
U
%7
~
PCTIUS2004/036297
WO 2005/042497
Yo
uX:
'tl
g
.1.
S'
0
u
0
......
'tl
Cl<
e=ll:
•
,
•
-.t
Vl
.....
u
.
.
'"
'"
I
I
..
'"
1
so
U
u~
.~ I ..
U
..
-% 7" .. :r:t ,..
Jj
<J
'tl
S'=ll:
U
'"
.....
-.t
I
PCTlUS2004/036297
WO 20051042497
r
I
0~
0)(0
".
]
0
so
U
':Z':z'
-r~
A
-
,J,
O~
0 ':Z'
0
'tl
, 'S-*
a-
~
U
I
§
,
2:~~.
0
o.
~.'
iS'
0
u
72"
.J,
0
'tl
S'*
U
-
It)
It)
.0
WO 2005/042497
peT /uS2004/036297
,;;
¢
nJ::
1S'.
J.
Q
U
0
..
"
. .
-
'l:l
g~'
~.
..
,
....
s,
..
'.'%
1S'
0
u
¥
%:x:
.t.
0
'l:l
S''Il:
U
~.',.-'
0
\0
....
V
.,
PCTIUS2004/036297
WO 2005/042497
Q
7"
't:l
g
l:l.
8'
:13
e
u
..
...
1",
.:
...
..
'"C:S
",.
t*
U
....
Ie
,
~
....,
-
'd
8
te
u
¢
rC:
.i,
n
U
'd
t*
U
<'l
Ie
....
PCTlUS2004/036297
WO 2005/042497
~,
nJ::
'tI
g
~
Q
U
i.
z:
b
"CS.
t:i
u
.~
....
I
....
M
o
~
:r
0
¢
't:l
~
a8
nJ::
J.
b
'CI
S'=lI:
U
v
10
.....
WO 2005/042497
PCTlUS2004/036297
t-
. "CI
l;,
'.
El'1l:
U
,
2!;
....
iJ
¢
"CI
~
~
u
TrJ::·
.t.
u
"CI
~'11:
U
10
10
.....
PCTfUS2004/036297
WO 2005/042497
~
~±T~
o~u
<J
't:l
§
_
0
:r:Jr
J.
0
S'
0
u
<J
b
#
."r
-
.
-
"C:l
$-
S'*
u
,
:8
....
,
1
~
u=
8-QU{~~=o
~ Q
:r
't:l
S'*
U
00
....
'D
PCT/US2004/036297
WO 2005/042497
1-
q'
~~J::
0
'g
6
8'.
0
u
)!.,
U
..
~
;;..
r-....
S'=Ii:
u
I
.
\C
~
~-
,(q
'0
~
S'
0
u
::0
. .zz
,~{
0
'0
0
~~
·0
....
t-
PCTfUS2004/036297
WO 2005/042497
t
L.
"*"
..
..
"Cl
§
0
~
0
U
(0
7"~0
.~(
0
.:.
"CI
a'*
U
'
M
..
l'-
rl
,
§
,
u
~ I
.~ ~
"
"=
~
S-e
Co>
,,:x:
.1.
0
"Cl
S'*
Co>
N
l'-
.....
<J
WO 2005/042497
J-
PCTlUS2004/036297
...
:r
U
I
Y
~
't:l
§
1-;;
".:zo
Q
.i.
S'
Q
C,)
0'"
0
0
-,
"Cl
S'*
C,)
.- -
'
. .-r--"'....
,
~
.....
,
1--"
':r'
u
"Cl
§
Q
~
C,)
•
~ '=''-1-''.
.l'
0
>d
S''Il:
Co)
"""
.....
l'
. U
:
WO 2005/042497
peT IUS20041036297
~
I
I
§
0
S'
0
u
~
~
A
~
..Y
~:x:
-r
.l.
0
,
.
.'
..
"
"Cl
f:ll=
U
'r-r-......
,
...
0\
o
I
Q
]
0
Cl<
e
0
u
'"'
8
QX
n
f ~
"Cl
S'=II:
U
r'"
~
WO 2005/042497
PCTlUS2004/036297
~
c!i
.
... ~
or
't:l
'%:x:
=
:;l
0
S'
'.
.1.
0
u
b
..
'd
,S:il:
'.
:
. !:::
C\
..
..
,
o
.....
,
..
8ti=o
~~,.
'l:l
g
aQ
or",
u
~,
,\)
. ..:;
'd
a-#
U
00
t-
.....
WO 2005/042497
PCTfGS2004/036297
l"
~
'"
¢j
u
",~A:
"l:l
~
S-
Q
U
.t.
c-o
a-~
"Q
.....
co
.....
U
I
......
.....
.....
I
"l:l
§
Q
S'
Q
u
'=X
0
Y..
.l'
b
'tl
~*
0
co
.....
<j,
':J:
WO 2005/042497
PCT/US2004/036297
't
y::
~
'"
"I:l
g
S'
Q
u
nJ::
,!,
b
..
~
t~
C'">
~
U
9
0
"I:l
~
t
Q
u
¢
:t%J::
,!,
b
'1:l
S':II:
U
<"l
~
PCTlUS2004/036297
WO 2005/042497
"r-
...
:r
<J
(;l~
"Cl
1:1
;:l
0
eel
u
:rzl::
,i,
u
..
"Cl
t~
"fri
....
ClO
U
I
~
....
I
"Cl
~
t
0
u
Q
0·'
~O
.
j,'
U
"Cl
S''"'
U
.r
co
.....
PCTlUS2004/036297
WO 2005/042497
..,.
0y~
~
""
't:l
§
0
.a0
u
771::
.t.
0
,.
.,
"c:l
S''!l: .
U
l"-
....
ee
PCTlUS2004/036297
WO 2005/042497
't"
9
u{:
"Cl
§
0
Qc
e··
.!.
=
u
U
.
...
....
,
,.",'
..
'
"C!.
t:il:
0\
00
..
.....
u
,
.,.,
.....
....,
y.
'"
'"
"C!
~
~
Q
U
nJ::
. c,
U
"Cl
~'#:
u
00
~
..
WO 2005/042497
PCTlUS20041036297
q
;r"
'1;l
g
:r.,.
0
S
0
:r
u
0
f ~
-
\
U~
'"
...
,
'I:l
~""
,
"
"
......
,...
0-
U
I
,...
,...
\0
((f
'iJ
'I:l
~
~
t.>
.,.21::
.i.
U
'tl
S'll:
U
0
0-
......
PCTlUS20041036297
WO 2005/042497
~
~
"d
§
0
.
.~
0
u
nJ::
.J.
U
....
- ..
"
....Cl\
1*
....
U
.
r....
.....
0
·ui
~:J::
. e.
.»
"Cl
i
t
el
U
b
"Cl
S'*
U
M
....
C7\
PCTlUS20041036297
WO 2005/042497
...
(j
~"
uJ::
1t
.~(
0
l;,)
0
"
','
';
,
,
:
"
"'0
'D':li:
l;,)
"
,
, "
I{)
'"......
..
~t
'0
~
t
Q
u
'0
t=ll:
U
~~J::
.£.
C
-.:t
.....
'"
PCTruS2004/036297
WO 2005/042497
..
<r
nl::
.i.
'0
§
-0
~0
C,)
0
..
'0
g,.
'
r-- -..
'-'
.~
~'lI:
.
,
...
0\
......
,
u
~~
"d
§
Cl
S'
Cl
u
ul::
J.
0
"d
S'*
C,)
...
\C
0\
PCTlUS2004/036297
WO 2005/042497
~
~
X:
"Cl
~
n
J.
t·
0
u
'.
"
b
.
'
,-
"Cl
'0-
0-.
~'iI:
,
§
,
.~~
::x: '
~
"c:l
§
Q
t
Q
U
.~(
U
"Cl
t'll:
U
':t
00
0-.
,.
PCTfUS2004/036297
WO 2005/042497
'f
n,{:
1
~I
"'Cl
=g
~
8
';t:Z-
(j
~2
P
V
..
"
"'Cl
~:Il;'
.......
0
N·
U
,
.....
~
I
I
I
.~
'"
¢/
:"j
<J
.
,{Q
"'Cl
g
~
Q
U
l'
'p
.V
"'Cl
~'*l:
U
0
0
N
0
-
WO 2005/042497
PCTlUS2004/036297
~o
1
~o
Q
t,)
uJ::
!,
Q
z
u
'
U
'C
t'll:
0<"1
0
N
t,)
,
s.
T...~
¢U;
.:(
...
'tl
g
f
Q
t,)
T%J::
)~,
P
,0
'tl
S''lI:
t,)
...
N
0
N
..
PCT/uS2004/036297
WO 2005/042497
9-.
nJ::
'tl
t
~
'S'
Cl
U
z?
2':r
Q
...
.'
U~
"/.,
'"
'"
'd
:g
~*
C'l
..
¢t~'r'"
\J
."
<.J-
nJ::
'd
~
S'
0
U
S'
.()",,:
'"
'l.
'"
'tl
S':tI:
U
'<!'
0
C'l
peT /US20041036297
WO 2005/042497
~
~\
UJ::
'tl
g
~
o-
Q
U
':l'%
r
-.
~'2"
P
(J~
:r
..
'l:l
.
'
.~
"
~:g,
N
,
~,
I
..
:r
u
¢l
?,'
.~
~%J::
't:l
~
S'
Q
U
,.~
P
<.1..
.:r
"Cl
'1:1<
El'*
u
\0
~
I
WO 2005/042497
peT IUS2004/03629 7
J?
_
v
.d
uJ::
"c:l
g
.-
-a-
ni
~~-o
0
u
p
:
.....
..
.
..
"c:l
0'\
0
N
~#
U
,
trI
~,
,..~
'-'
¢l
I'
T~i:
"c:l
§
~
>,,c
U
\-0
II
_
0
'l:l
S':Il:
u
00
~
PCT/uS20041036297
WO 2005/042497
y,
n"c:
,,{
'tl
§
Cl
" c..a
Cl
\,)
P
o=-\
9
'"
"C
"r=ll: '
,
....
,
~
\,)
g~
~
'"
v
~
"
'7,'
"Co.
"Cl'
~
~
8
T%
i
p
~:r
'%'2
0,,"":\\
0
"Cl
S':lI:
\,)
....
N
0
~
,
wo
PCTIUS2004/036297
2005/042497
;-
I
(j
'1:l
§
0
S0
u
''9
~:J::
e-.!
U
..
'
.
.,
'tl
t~
U
-
. 'M
C'l
I
~
."...,~.
<y
~~A:
."-1
..
,?
. ~.f
§
0
S-o
u
0
t*
U
N
..-.
N
i.
PCT/US2004/036297
WO 2005/042497
;~
7:X:
"d
§
,1
0
~
U
...... '%
~2'
0
.,
.'
'.'8, ..
s*
U
'0'
N
,
00
N
,
.JI
]
e0
u
nJ::
,~,{
U
"d
~*
U
"<t
N
PCTlUS2004/036297
WO 2005/042497
~
,...
<.J
~l
,1,
"t:l
0
~
,
aCl
u
,.:z~o
0
,
.
1. ,"
s*
," l--
'
.-<
C'l
U
.
.
~
.-<
-9
Ct
o
.;.:;
"
iij
"l:l
~
a-
8
c,.{
0
0
"t:l
S''!l:
U
I£)
.-<
C'l
WO 20051042497
PCTfUS2004J036297
..
~"
0
'l:l
~
~
0
,./~o
,j.
0
U
'.
'l:l
. ~~
U
..
.
".0'>
.
N
.
-.
o
<'"l
~.
<J
~
0
'l:l
g
~
0
U
:<"~Q
.j.
0
'd
~~
U
...
00
N
peT IUS2004/036297
WO 2005/042497
-y,
u
u
0
iS'
n~o
0
c.>
.~.
U
•
"C
I'
••
=- .
s=!t:
U
...
. .-f'
~
,
.....
.
M
.....
u
7'
~.
1S'
0
u
0
~%Ao
.j.
0
"C
S'=!t:
U
0
81
PCT/US2004/036297
WO 2005/042497
~
2:1~
:r
0
lJ
~
't:I
§
0
0
':r'%
,c..
I
.£.
. El .
Q
U
0
..
"
-
.
..
'C.
~~
",'
""~
,
~
....
,
'C
g
S'
Q
U
.1.
~
b
i~
Co>
""
C'l
N
0
PCTlUS2004/036297
WO 2005/042497
N
<J~
:r
'0
\
.
$
"'''' 0
.J.
~
aQ
Co)
0
..
..
.,
:
'l:l
.g"
El'll::
U
"I:::l
~
e-
o
u
r--'
N
.N
r
.1. '
0
'l:l
~=Il:
U
~
N
PCTlUS2004/036297
WO 2005/042497
~
,2
"C:I
~
S'
.~(
Q
U
b
..
"C:I
..
...,
N
t=ll:
0
U
I
I
&"
.2.'
I
o~.
o;'U
o·
"C:I
~
t
8
0
b
"I:l
u~""
00
f;l
"':r'"
WO 2005/042497
PCTlUS2004/036297
'"
.£ .
:r
U
0)
"'.............
0 ~
IA
T
"CI
T
0..........,'"
§
t·
0
u
o'
J,
":E'Z'
b
'.
'C
a-=Il:
U
"''""
C'ol
,
\()
~,
2/
.-1'
.., .
."..
.
~
't:l
§
Q
t
Q
U
T" .
"CI
t'll:
U
'0
1,
b
""'~
(J'
I'
(?'
WO 2005/042497
PCTlUS20041036297
b
,1
'~'~
'tl
·S
g~
e
U
".'"
0
.i.
b
'tl
f=ll:
10
N
'"
U
,
10
c<1
......
I
~
~
".
(j
".
,0
'x'
I.
I
-b
'Cl
~
a-
e
i,,)
,I,
~%
b
'Cl
a-=Il:
U
'"
~
0
PCT/US2004/036297
WO 2005/042497
~'
n1::
~
§
0
s-
o
U
.1.
~
o~
:r
V
-.
";"'.
,
""
"l:l
.S''#:
00
'"
N
U
,
r--
~
I
~
q"
:r
U
·1"
'~
~
§
'7,,1::
Q
S'=
i=
(.l
." .
Q
<.J..
:r
"l:l
S''#:
U
P
r--
~
i
peTIUS2004/036297
WO 2005/042497
~"
%%1::
1
1
'S'
el
u
iF
~~ ~...
,
..
,
.:
..
"
.=,,'
a':II:
."
0
v
M
U
,
00
.....
....,
qi
.,~
<J
n1::
."
t
~
a~
u
;F
u
.,..
<..I....
.
~
"Cl
S':II:
U
'"
lQ
PCTlUS20041036297
WO 2005/042497
~'
nJ::
16'.
..
Q
U
~
.-q"
..
"
r~'.
'C
..
,
.
:
,
.
..
"
' ,"
~
N
U
,
'"
~
I
","
<)
<J'
,
~I'
'C
~
S'
Q
U
T%J::
1
"'%
.-q"
~%
L.
'C
S'=Il:
U
.....
~
".
<J
WO 2005/042497
PCTIUS2004/036297
~
Vo
nJ::
~
~
S-
o
U
":1'%
i
....,,;;:Z'
])
It;
:r
..
.
"d.
-;
f~
N
U
I
~
....
I
..
¢J
"0
~~;(:
1S'
t
0
u
J3
%
"c:l
S''It:
u
""
~
WO 2005/042497
PCTlUS2004/036297
>-
Y-.
%%1::
'C
=
'S'c
::I
c
.J.
u
.
"
..
..
.
"-9
..
,
..
'Cl
\0
1=11:
'<T
'"
U
,
rl
;:!;
,
<r~
:r
...
?P". ,
~.I
'C
:r./(:'
§
0
i
u
.i_
"-9
..
"Cl
1=11:
U
~
'"
PCT/US2004!036297
WO 2005/042497
~
I
~
"
~O
;.... ~ I
'1:l
~
~
Q
U
",,,,~o
(J
,,'"
J,
0
.,
.
,
~
t=ll:
01
.
~'
U
,
~,
i
Q
t
Q
U
f
i.·
...
,
U
.
~ ~
'1:l
~=II:
U
r--
~
WO 2005/042497
PCTlUS2004/036297
)-
~
:r:or
'0
§
n~o
0
.i,
i
0
u
0
..
'
.
!
,'0
S':lI:
.'
..
....
.,
--
~
U
,
~
I
'0
~
~
u
R
.
.t.
:r%
b
'0
S':lI:
U
0
~
u
0
peT IUS20041036297
WO 2005/042497
9'~
"''''
"C
§
0
'S'
0
u
n~o
i.
=r.
0
"C.
C'"l
~
~=lI:
U
.
.
~
....
y
i
I ..
.&
"C
~
S'
0
u
":I:::r"
. c.
".".~o
0
"Cl
S''**'
U
C'l
'"
C'l
"
peT /US2004/036297
WO 2005/042497
j
~
"
(J
~t
"Cl
2":
m0
g.
s'
0
U
%~
"'''
0
'.
,
.
,
.,.,
on
"Cl
S'*
U
'N
,
~
I
7"
I~
"I:l
g
r
u
%"
n~o
.J.
0
"I:l
S'*
U
~
lI'I
C'l
WO
PCTfUS2004/036297
2005/042497
"'
¢u~
."
<J
(0
"Cl
§
Q
.,,:r~o
~
,f~
Q
u
0
..
"Cl
.•
&.
S'~
U
Ii"l
N
"
~
.....
,
§
Q
e~
r~¥
o ·
.
z
'.
''%
• -
";I;
2:~
7%
-0....
"Cl
S'*
U
"K :#
.' .. ~~
00
N
'"
PCTlUS2004/036297
WO 2005/042497
~
0)
'l:l
§
1?
Q
~
8
0
"
,
...
",
'l:l
"
I:l<"
"CJ
......
:
"
\0
N
=II:
,
co
V
......
,
I
9L
<;l
'l:l
~
a0
U
2;::
:r:~'
%'%
0
'd
a-=II:
U
<:>
\0
'"
PCTfUS2004J036297
WO 2005/042497
I
a
V
"0
~
~
Q
U
5::: P
7'/
:l!":t:
0
,
..
.'
.'
=-., .
El'll:
"0
-""
'D
N
U
.
.
'"
::!:
1
~
Q
u
. ·/v
o:()
~O
",-7'
"':
U
d
"0
S''!I:
U
N
'D
N
'~
PCTJUS2004J036297
WO 2005/042497
~
:r
~U
O~
~~
1",
'0
.g
S'
,L
0
u
0
..
..
...
'0
-.
lJ'\
\0
C'l
~=lI:
U
I
~
.....
,
~
1
~
0
u
:r:J::
.f~
0
'tI
a-=lI:
u
..
'&
:z
N
'u
PCTlUS20041036297
WO 2005/042497
~
:r
U
¢/
-=
g=
:r:rJ:
O
,~
c
u
./.
0
',0
,
'tl
'0
'0
,
'
:
"
or--
S':#:
\0
N
U
--.
ltl
.y ~
~
'tl
g
S'
Q
u
",:r
.1
0
-=
S'*
U
\0
\0
C'l
WO 2005/042497
"Cl
§
0
~
0
{,)
PCTruS2004/036297
'''/Il\
.~..
"
U
,
I'
~.
'
0'1
t*
U
\D
C'I
\..{)(v
"Cl
~
s-
o
U
1\0
%.
__'Z."Z
0
't:l
S'*
U
00
\D
C'I
.
peT IUS2004/036297
WO 2005/042497
,q
'C
5
Q
.~
Q
U
..
.r,
"
~
'.
b
.,
..
'Cl
'.'
.r::
1=11:
C'l
u
;(r~
'Cl
§
t
Q
U
,1
.{
b
'Cl
1=11:
U
0
r-C'l
"
+
peT iUS2004/036297
WO 2005/042497
£'
yo"
b ~
.
'Cl
§
.~
Q
~o
~
I
.»
Q
U
%
l~
..
..
t"-
on
N
U
.
~
....
,
~
¢ri
%
<.J
.~.
'Cl
g
1
o~
.
."2
b
'Cl
f=lt:
U
\D
on
N
PCT/US2004/036297
WO 2005/042497
...~
OU
o
~
,1
'C
§
0
'S'
J~,
0
u
b
.
. -.
"
.
.
'.
. 'C.
en
S''ll:
N
u
,
.,jo
>n
.....
I
~
".
U
I
o~~
'd
m
S'
u0
,i
,.(
b
't:l
S''ll>
U
N
~
WO 2005/042497
PCTlUS20041036297
2:'
,.1 "',
't:l
g
~
.~.
Q
l;,)
b
.
,
,
"
't:l
't=ll:
l;,)
'
'
" ,,'C
~
,
trl
trl
,.....
,
PCT/CS2004/036297
WO 2005/042497
I
I
I
2
Q
%
'0
I::l
::
(;)
..
t·
c
U
..
.J. '
U
,.
:
. .
'0.
t*
U
00
!'
N
,
~
....,
or'"
<J
'0
g
tc
u
.i~~.
~
b
'0
S'#
U
!'
M
PCTlUS2004/036297
WO 2005/042497
>
§
<:>
j
<:>
U
21
..~
"Cl
..
. .
~" ""
.J. .
..
U
..~ ..
.
. .
. "Cl
S'*
U
0
co
<'l
I
<i
¢
'l:l
g
S-e>
U
7~'~:'
./-
0
'l:l
t*
U
<:1'
N
peTIUS2004/036297
WO 2005/042497
>
~
~
,..
~
,..'"
"Cl
~
0
'.
~
u
'tI..
,..
.t.
0
b
,.
.., .
,
"Cl.
a-~
U
.<'1
00
<'I
I
00
:!:l,
,..'"
<J.
¢
~.'
"Cl
~
a0
u
,..~rC:
J.
0
"Cl
S''Il:
U
00
<'l
T
WO 2005/042497
PCT/US2004/036297
)-
''<l'
.~ DO
I
0-
....,
V'l
§co
~
Q
U
WO 2005/042497
PCTfUS2004/036297
~
X
<.J
{'y
I
"Cl
§
,%%J::
0
S'
.t.
0
u
..
"
0
....
.
..
"Cl
t=ll:
U
"
,
..
'"
"
'00
N
,
§
I
%
Q
"Cl
g
S'
Q
U
UJ::
.;C.
0
"d
11')
~=Il:
U
00
N
PCT/US2004/036297
WO 2005/042497
.....
v
n~o
¢
"Cl
§
0
S'
0
u
.
,
.'
.T~J::
.1,
0
'"
'd
9'*
'00
DO
N
U
I
.....
~
~
:r.
'u
f
¢
"Cl
§
0
S'
Q
u
T~J::
.1,
0
"Cl
u~*
to
C'l
;
WO 2005/042497
PCTfUS2004/036297
I..
"*
(0
't:l
§
0
t
0
u
,.=~o
J,
0
-,
l*
..
0
0N
U
I
fi§
....
I
...~
<J
X
,0
.~~
't:l
A:
g
f
u
-p
I.-
~
0
n
..(.
0
-
't:l
~*
U
Cl\
CO
N
PCTIUS2004/036297
WO 2005/042497
v
Q1/
0
~
~
TT,,(:
,a-
,1,
8
0
"
.
:..
'
'.
'1:l
,
. . .,
"
'
..
..
~.
S':tl=
~
U
I
m......
I
0"
n,,(:
'~
~
~
t
0
U
I
"
'r:i~
"'T
0
~
~'lI:
U
......
~
(,,,,,
PCTfUS2004/036297
WO 20051042497
.~
'tl
~
~
e
j
00
U
~%X:
%?'
z":r
0
'tl
t*
U
..
o'
-to
~
,
$
....
,
I
I
~
§
e-o
0
u
UJ::'
J.
0
'tl
r<l
~'II:
U
~
PCT/US2004/036297
WO 2005/042497
..
~f'
nJ::
"l:l
~
·f
.i.
0
u
0
,
,
,'.,
'J'.
6'*
U
.
\0
C7\
C'l
I
§
,
~
T
9
~%J::
?
,-
~I
1S'
0
l;.)
.i.
b
"l:l
S'*
U
V1
C7\
C'I
WO 2005/042497
PCTlUS2004/036297
~
".
U
I
::::,..I
~
1
Q
f-
8
.'
Q ....
</,.
".
0
.".%1:: _
.!.
0
'1. .
ao
El=ll:
0\
N
e.>
u
¢Y.
't:l
§
Q
~
e.>
.
"."
~
'"
uJ::
1.
b
't:l
t=ll:
e.>
l"-
g)
~
PCTruS2004/036297
WO 2005/042497
..
,..
u
I
:
~
"C
~
~~
0"
UJ::
S-o
u
-0
"
;-~-
I
.1.
b
•
4
......
"Cl
U
0
0
....,
I
~
.....
,
1
~
u
,..zJ::-
.1.
0
"C
S'''*'
U
0C'l
'"
PCTlUS2004/036297
WO 2005/042497
~
'"
<,)
(~
:::,...
<,)
-=
.",a;::
~
t
.£.
Q
u
0
.. ,.
'. ..
.
. ..
:;,'.,,,i-t ..
\:1
1:"",.
.....
"Cl
.Q,.
~
El'*"
'"
u
,
-
~
I
"Cl
~
t
Q
U
-=
a'*"
U
r8
~.I. o /)rI~
. ..
.;:r
":0
0
•
~ ~
-
0
<'"l
PCTIUS2004/036297
WO 2005/042497
9YO
~
~
nJ::
"Cl
§
0
~
0
,J=
u0
'0
'
:
.
..
"Cl
~
'~:lI:
«)
U
,
0\
\0
.....,
..
.¢Y
%
1
a0
u
uJ::
j,
b
"Cl
S':lI:
u
«)
0
«)
15
0
WO 2005/042497
PCTIUS2004/036297
..
u
~.
u1::.
1
J,
~
u
...
0
.
:.
,
,
-=
S'~
10
0
<"l
U
,
g
......
I
tf
.0
~~~
.
..
0
1S'
~
, ~o
zz·
u
~,
. ~
,\)
.1 .
-=
S'~
U
In
0
<"l
.,&'
peT IUS2004/036297
WO 2005/042497
c;r'
T
~I
~
nJ::
]c
!'
c
.1.
u
0
..
.
:
.
'l:l
r».
. US'*
0
r<l
..
..+..
·19
'l:l
g
S'c
u
T2;(:
.~(
0
'l:l
S'*
u
8
<'"I
PCTIUS2004/036297
WO 2005/042497
)-
~
n'c:
'e
§
Q
'.
J.
'~
Q
U
b
'
..
..
"
'e.
0
......
~""
<'l
U
...~
<J
0""
{q~
'b
~....
'e
g
f
u
".
"J::J::
.i.
b
'e
S'''''
U
<;I
0\
0
'"
PCTlUS2004/036297
WO 2005/042497
tuQ
'tl
~
i
=
u
::J::
.i~
0
.
'
.. .,
"d
.
~*.
:
~.
""
..
.~ I ..
~
1
i
=
r..>
A:
u
J.
0
"d
S'*
U
.....
.....
<"l
;
WO 2005/042497
PCTlUS2004/036297
Q
.y
Ih-
'tl
g
i:
S'
'U
Q
U
.~(
d
..
..
..
'tl
S':ti:
U
.""....
"".
,
....""
......
,
..
oX"
¢"
'tl
g
S'
Q
u
'T.%.i:
.£.
b
'tl
a-=ll:
U
....
'"
<'"l
peT IUS20041036297
WO 2005/042497
s~
'CI
~
t
Q
u
.
-
h,
..
uJ::
.1.
b
-.
'C
Q,.
a
u
=II:
.....
'"
'"
,
111
!:;
,
OJ?
1
S'
I:)
u
uJ::
,j.
b
'C
S'=ll:
U
.....
<'"l
V)
peT IUS2004/036297
WO 2005/042497
•
00"
.....
•
<'1
I
\0
r-.....
§
0
t
Q
u
(J
s ~~:o
"
!J
~
"
,"
o~
,.
'l:l
t'!l:
U
r-rl
<'1
"
PCTfUS20041036297
WO 2005/042497
I
~ "'1
9l
i:'
~o
if
':r'"
.i.
0
u
0
.- ...
0
.'
..
'.
...
. t:jj,
'C
.'
~
<'l
U
..
,
.
r-
.
~
u
~o~
1
~
0
u
':rzio
.i.
b
'C
S'=Il:
U
0\
'"
wo
PCTlUS2004/036297
2005/042497
I
~
<J
'~~
'1;l
§
0
c..
,~
u
~%J::
<J..
""
.i=
u
"
,
,
"
'1;l
N'
8''11:
N
'"
U
,
.
00
l"'-
.....
..
<:J'
~l
1:0.
.J.
I '
'1;l
§
0
~
u
2:#
.
d
'1;l
S''II:
U
.....
N
t"l
Q
WO 2005/042497
PCTlUS2004/036297
qt
..
CO
"C:l
~
:r".~o
.t.
.~
Cl
u
0
:
"
.~
~
N
a=ll=
"".
C,)
,
0\
r--
.....,
<Ii
¢r'
"C:l
g
Sel
u
%Z1:
.t.
d
"C:l
~=Il=
C,)
M
N
""
PCTruS20041036297
WO 2005/042497
?o
,q
--.:::
l,q
o
1S'.
0
C)
..
"
7~~O
,,C,
0
"
.
.,
'l:l
"
\0
t*
M
..,
C)
,
o
00
,
<oJ.
·:I.~
;~
.
:z ",
'l:l
§
Q
t
Q
C)
:=~:
.t.
U
'l:l
S'*
C)
Irl
f;:l
,....
u
WO 2005/042497
peT IUS2004/036297
.*.
ui::
u
"C
=
0
==
I:l..
·s0
u
..
.1,
0
.
..
'
"C ..
t=ll:
00"
N
'"
U
"C
~
t
0
u
ui::
.J.
0
"C
S'=Il:
U
r--
N
M
,
PCTlUS2004/036297
WO 20051042497
..
'"u
oJy
1a-
0
C,)
u~:
i.
0
.
..
...
..
..
.
.
"c:I
~
. 8 '*
C,)
0
.~
,
C'l
....,
00
I
C,)
PCTlUS20041036297
WO 20051042497
01
~'y
-:PI
~
i
l:l.
Q
n
J-
u
,
:
i:
,
's
b
~
l:l.'
~'=Il=
<'l
,
,
C"\
'"
,
C"\
00
'"",
8
l':.~ ))
'~
IJ
0
i
~
>=0
,..,..
Q
u
~.
'\,
~
~=Il=
u
....
'"'"
PCTlUS2004/036297
WO 2005/042497
1f'
..
Q
u
.., .'}-J,-8'
cr>-~~
..
..
.
~.
J:l<
.
B:#:
,
-.t'
.....
<"l
U
,
~
,....
I
I
~,.
~
~1:1
5
t
Q
U
.T%J::
J,
0
~
ut*
""'""'
""'
WO 2005/042497
PCTfUS2004/036297
.;'
<.J
I
o~
0
)l
.&
%
"
"Cl
'J
7:~
UJ::
g
.S'
,J.
Q
f,)
b
.
~',
,
"Cl
."
,.
..
,..,,..,
'D
S'=Il:
U
,
It">
co
.....
,
9'.
.':l'
0
"Cl
g
S'
Q
f,)
':zl:.
./.
b
"Cl
S'=lI:
f,)
,..,
V'l
'"
PCTlUS2004/036297
WO 2005/042497
1'"
'Xz~a
¢
'0
§
UJ::
0
t
./.
0
u
b
"=
Ii .
U
=II:
co
"
e<!"
e<!
,
-
\D
00
,
ft
'J:0'9"
"I~.
"C
§
0
S'
0
u
o.
;r"Z"',
J~
b
'"6'=11:
U
.f'
le<!
'"
PCTlUS20041036297
WO 2005/042497
8-
i
f ~~ 0
0
: .y
~
U
....
.
~~=o
.
..
"Cl,
S'=ll:
U
:r
0
v
""'
I
rOO
......
,
"'y~ '.~
. I
<[00 ~
i
~
0
u
<r",l~
.i.
b
"Cl
S'=ll:
U
0M
M
PCTlUS2004/036297
WO 2005/042497
Qn
oJy
"Cl
:0=s
~%,;(:
e-o
.i.
C,)
'.
.
b
,
. ...
'
"Cl
t~
N
..
'<t'
crt
C,)
,
CO
CO
,.....
..
~
.',n. ',;~..
.~
"CI
g
~
U
Q
'7%1::
.iT
0
"Cl
S'~
C,)
,.....
'<t
l"'l
PCT/US2004/036297
WO 2005/042497
o~ ~
¢
]
'S'
uJ::
0
0
t,)
.1.
0
..
:
.'
~.
"
'd
t=ll:
"
..
~
C'1
t,)
,
a..
.
....
QO
9-tr
,.(0
.
'\ OJ
%~:r
'd
~
s=
u
'%Z.
0
.1.
0
'd
S'=ll:
U
..,
"<t
C'1
peTIUS2004/036297
WO 2005/042497
...~
'J
~5"
6
n~:
'tl
§
0
a-
.1.
p
0
C)
..
o.
.0
0"
'tl
00
\()
-.r
"S'=!!'
..
'"
C)
,
o
~
,"
~" ~o'
"0
%%~:
't:l
~
i
u
J~.
..~
'tl
a-'Il:
U
II">
-.r
'"
'~
PCTfUS2004/036297
WO 2005/042497
.
..
¢rc
n!:
]
.1,
.~
Q
u
¢
..
'.
't:l
c..' .
~.
.....
S:Il:
U
:
~
'"
<J
.-
:~
~,
~\J.
't:l
g
0
o::{
s-o
.. '"
Co)
S
~I
~
"Cl
~=II:
Co)
~
.....
..
..
wo
PCTfUS2004/036297
2005/042497
I
~u
.~J::
'0
§
Q
a-.
Q
u
,1,
¢
~
'0.
0
~."lt<
.'
II'>
<'1
U
.
~
I
..
'"
<.J
"*
~,(:
.-T
<.J'~I
~,
.-
<.J.
]
Q
aQ
U
.
,j,
¢
..
'0
a-'!I:
U
0"<t
""
:
PCTlUS2004/036297
WO 2005/042497
~~
i:
'tl
:0=:s
.t.
n
S'
0
c.>
P
l.
,
..
,
'. . .~
"
'tl,
'S'*
c.>
".
<'l
\(')
t"I
0J9
1:0
~, , »
'Cl
g
a0
u
.1.·
Q
..
'tl
~*
U
-
\I')
rr'l
peTIU52004/036297
WO 2005/042497
t~t
"I.,
i:
:r
't:l
§
T:
Q
.,
~
u
J.
9
..
.
".
~
-v
If'l
~=Ii:
'"
U
,
"t.
,...;
,
~
¢/
T'
U
l:
:
b
'T~J::
't:l
~
i
u
.j~
~
!-
't:l
~=Ii:
U
'"
~
,
PCTIUS2004/036297
WO 20051042497
..
~
:r
~
'ti
§
0
S'
0
u
0
:r:J::
.J.
0
....
,.
"C.
t=ll:
\0
It'>
<'"l
Co)
..
~
t
0
u
.t.
n
0
"C
S'=II:
U
~.
X:
"C
~
.
It'>
\f'I
<"I
:r
5i.
peT /uS2004/0362 97
WO 2005/042497
)
r
..
«
§
0
.
'a-
'
0
u
.
%%J::
J.
0
,
"tl
a-*
'
00
"
l(")
<'l
Co)
,
\0
'"
'"',
..
....
. '1::>.. .. ,
~~i:
.'
~
1f
e
u
,I.
0
"tl
a-*
U
,.,..~
l"V">
<"l
0
WO 2005/042497
peT IUS200410362 97
)
y
"u:;x:
1S'
0
u
nJ::
.i,
0
'.
..
~
t''ll:
U
-0
'"
<T'l
,
.
l"
'"
,....<
v'
~~J::
"Lo.
"l:I
~
t
=>
u
.....
.i.
U
"CI
S=ll:
'"
V"l
«)
•
WO 2005/042497
PCTruS2004/036297
)C(C:
't::l
g
0
~
0
u
..
nl::
.i.
b
..
't::l
.
,
"
·N
\0
<'l
~:II:
U
,
co
~,
'x(t
L.'
iS0
u
.
UJ::
.J.
0
't:t
S-'#:
U
·.D·
.....
\0
<'l
L.
PCT/US2004/036297
WO 2005/042497
':
ly
0
".."' .....11
'C
g
S'
Q
U
o ::::v.- '" '",
-r.,. ~
~ ~
-
..
'C.
\0
\0
f'*
'"
U
I
a,
a,
,
..
,
'~¢(
.
.'
. ..&
1
<:l
~
u
T%~:
.l.
b
'C
S'*
U
<'"I
\0
<'"I
WO 2005/042497
PCTruS2004/036297
'!"
ly
0
"..%....
't1
§
0
11
8
'S'
",
0
u
~"
"
i~
u
..
'.
ffl
....
,
o
~,
y
0
"!f
".." ......1/ '
ti
~'
't1
§
%~'%
Q.
0
S'
0
u
""" ~
~
'~
(j
'l:l
S'*
U
r-.
....
\0
PCTlUS2004/036297
WO 2005/042497
,
~
.0'
"i~ I
o
1t
Q
u
.
...
8'
'
.,.~
-
= ~
u ....
f. ~
'tl
~*
$
<'l
..
.4-.
PCTlUS2004/036297
WO 2005/042497
[00106]
Although certain exemplary embodiments are depicted and described above and
herein, it will be appreciated that a compounds of the invention can be prepared according to the
methods described generally above using appropriate starting materials by methods generally
available to one of ordinary skill in the art.
4. General Synthetic Methodology:
[00107]
The compounds of this invention may be prepared in general by methods known to
those skilled in the art for analogous compounds, as illustrated by the general schemes below,
and the preparative examples that follow. Starting materials are commercially available from
typical chemical reagent supply companies, such as, Aldrich Chemicals Co., Sigma Chemical
Company and the like. Compounds that are not commercially available can be prepared by one
of ordinary skill in art following procedures set forth in references such as, "Fieser and Fieser's
Reagents for Organic Synthesis", Volumes 1-15, John Wiley and Sons, 1991; "Rodd's Chemistry
of Carbon Compounds", Volumes 1-5 and Supplementals, Eiservier Science Publishers, 1989;
and "Organic Reactions", Volumes 1-40, John Wiley and Sons, 1991.
Scheme 1
[00108]
Scheme 1 teaches the general preparation of compounds of Formula 1. Typically
co~p,?unds of Fonn!1la I, where Y or yv is
":"
-AI( ( >;~'h
o
wherein A is -NH-, or -N(Cl-6alkyl)- are prepared by the coupling of optionally substituted
compounds of Fonnula A, which has a nucleophilic function with optionally substituted
..
'
.
;.,
.'
.
'.
..
'.
compounds of Formula B. which have a terminal electrophilic functionaIilty, such as a
carboxylic acid. sulfonyl halide, isocyanate. or the like, as defined previously. These methods
are also applicable to compounds of Forrmulas II and
- 202-
m as defined previously.
PCT/US2004/036297
WO 2005/042497
B
y
1 x=
/
OH, OR, halo, leaving group
(RA)p:)-Aya < }(R~'
o
- 203-
PCT/US20041036297
WO 2005/042497
Syntheti}: Scheme 2a. 2b:
[00109]
Scheme 2a teaches the preparation of optionally substituted benzimidazole
compounds of Formula II.
Scheme 2b teaches the preparation of additional optionally
substituted benzimidazole compounds of formula II.
y
(A')ce"H
,
I
+
NH2
j
Scheme 2a
- 204-
WO 2005/042497
PCTlUS2004/036297
~NH2
~
NH2
1) Acylate, cyclize
2) Deprotect
j
PG
HO~~
+
(RA)r-~
PG = protecting group
A
x = OH, OR, halo, leaving group
,II
'.
S~heme·2ti·
An optionally substituted l,2-diaminobenzene
is reacted with an optionally substituted
carboxylic acid with a protected nucleophilic group to provide the benzimidazole
which is then cyclized to form the benzimidazole
and acylated, . sulfonylated,
carbamoylated,
. .,
.
moiety. The nucleophilic
intermediate,
group is deprotected
of Formula ll.
. or alkylated
.,... to provide compounds
.
...
- 205-
..
PCT/uS2004/036297
WO 2005/042497
Synther!!c Scheme 3a. 3b, 3c:
[00110]
Scheme 3a teaches the preparation of optionally substituted aryloxy acids. Optionally
substituted aryloxy acids are prepared by reacting optionally substituted phenolic compounds
with optionally substituted halo-substituted alkyl esters (X is el, Br or 1) to obtain the
corresponding ester.
3
Scheme 30
The ester compound is then hydrolyzed to obtain a desired optionally substituted compound of
FonnulaB.
3
B
Scheme3b
As taught in Schemes 1, 2a, and 2b, compounds of Formula A and B are reacted together to
obtain compounds of Formula I. For example, FonTIulaA as taught in Scheme 2b and Formula
B as taught in the present Scheme 3b can be reacted to form the corresponding compound of
. fOI:Qlula I (Scheme ~c). .
-206 -
WO 2005/042497
PCT/US2004/036297
SCheme3c
Synthetic Scheme 4:
[00111]
Following the procedures taught in Schemes 1, 2a, 2b, 3a, 3b, and 3c, and using an
optionally substituted aryl isocyanate compound of Formula B,
0r
(AAlr-V--}-1
.
N
.
,~.,\. ' A
. y'
H
"1'"
O==C===N-R'
.
......
Scheme 4
optionally substituted compounds of Fonnula I are obtained.
Synthetic Scheme 5:
·207 -
B'
.
PCT/US2004/036297
WO 2005/042497
[00112] '>'- Following the procedures taught in Schemes 1, 2a, 2b, 3a, 3b, and 3c, and using an
optionally substituted aryl sulfonyl chloride compound of Formula B,
B
(R
A
1
l-A-S R-Q(AI)S
\\ /;
II
o:~
N
)'-1
~
z
\
y
0
Scheme 5
optionally substituted compounds of Formula I are obtained.
Synthetic Scheme 6:
[00113]
Protected optionally substituted benzimidazoles of Formula A are prepared by
reacting a star4ng cy,'!Jl~s~bstituted benzimidazole ~~th (BOC)20 followed ]:>y reduction of the
~r.thel~e): t\?op~ovjdethe.4esii'ed·~o~poWld.:pfForm:uiaA,.'
. c~~o' ~oup (R~ey':1"UckeIlfJ;2
,.
...
.' ..... ~,... .
"-":::::
(RA)
Ct
I
&=
H
N
~
N
CN
..... ~:"". "0 '
A}-O
(R lro=-..:::::
•
1
A
N
k .
/hCN
N
. . 6' .':,').(;
·
t-
0
A
(R )rcr-..:::::
I
N
N;,'>-----.\.-NH2
A
A
.scheme 6'
As taught previously in Schemes 1, 2a, 2b, 3a, 3b, 3c, 4 and 5, compounds of Fonnula A from
the present Scheme 6 are derivatized to provide the corresponding compounds of Formula 1.
Synthetic Schemes 7a, 7b:
- 208·
WO 2005/042497
PCT/uS2004/036297
)--
[00114]
An optionally substituted benzyl alcohol is reacted with phosgene to provide an
optionally substituted compound of Formula B,
B
Scheme7a
This compound is then reacted with a compound of Formula A to provide the corresponding
compound of Formula I. For example, where the compound of Formula A is a benzimidazole
taught in Scheme 2, the following compounds of Formula I, where s is 0 to 4, and m is 1 or 4, as
it corresponds to Q where Q is -O-Cl-4alkylare obtained,
+
B
A
J
.. "0:": ~~Ay'.'....
J~r~
. ""
.. "" ...'
~
"z '
\
y
,
0
Soheme7b
Syntheti~ Scheme 8: ,
[00115]
A compound of Formula I, where W is
-209 -
o~
.'
n·
'~R~~"
:~.-.~
(CH~m"
..'
... ,
",
"
PCTlUS2004f036297
WO 2005/042497
is prepked by reacting an optionally substituted di-amino benzene with an optionally substituted
benzaldehyde or benzoic acid to obtain the corresponding optionally substituted compound of
Fonnula I, where RA, RD, r and t are as taught for Formula I, in a manner analogous to Schemes
2a and2b.
[00116]
Although certain exemplary embodiments are depicted and described above and
herein, it will be appreciated that a compounds of the invention can be prepared according to the
methods described generally above using appropriate starting materials by methods generally
available to one of ordinary skill in the art.
5. Uses, Formulation and Administration
[00117]
In yet another aspect, a method for the treatment or lessening the severity of acute,
chronic, neuropathic, or inflammatory pain, arthritis, migrane, cluster headaches, trigeminal
neuralgia,
herpetic
neuralgia,
general
neuralgias,
epilepsy
or
epilepsy
conditions,
neurodegenerative disorders, psychiatric disorders such as anxiety and depression, myotonia,
arrythmia, movement disorders, neuroendocrine disorders, ataxia, multiple sclerosis, irritable
bowel syndrome, or incontinence is provided comprising administering an effective amount of a
compound, or a pharmaceutically acceptable composition comprising a compound to a subject in
,ne~d thereof. ,In certain preferred embodiments, a method for ,.the trea~ent
: ,s~verity :of,':acute,,' 'chronic, neU,ropaWc', or' infl~t~ry
pain'"
OJ:
lessening the
is: provide.d" c~mpriSing ",
an eff~cti~e"~J~lOunfof a -cQmpoun:d:~; '& phannaceutlc8ny
'aChiiinistermg
'acbeptable CO~pd~iiion''.
to a subject in need thereof, In certain embodiments of the present invention an "effective
amount" of the compound or pharmaceutically acceptable ,composition is that amount effective
for treating or lessening the severity of one or more of acute, chronic, neuropathic, or
inflammatory pain, epilepsy or epilepsy conditions, neurodegenerative disorders, psychiatric
.".
.
.".
.
..',..
.
"
..
'
disorders such as anxiety and depression, myotonia, 3.1Tythmia, movement disorders,
neuroendocrine disorders, ataxia, multiple sclerosis, irritable bowel syndrome, or incontinence,
[00118]
As described generally above, the compounds of the invention are useful as inhibitors
of voltage-gated sodium ion channels or calcium channels, preferably N-type calcium channels.
In one embodiment, the compounds and compositions of the invention are inhibitors of one or
- 210-
WO 2005/042497
PCTfUS2004/036297
ll;
more of NaVl.I, NaVl.2, NaVl.3, NaV1.4, NaV1.5, NaVl.6, NaVl.?, NaV1.8, NaVl.9, or
CaV2.2, and thus, without wishing to be bound by any particular theory, the compounds and
compositions are particularly useful for treating or lessening the severity of a disease, condition,
or disorder where activation or hyperactivity of one or more of NaVl.l, NaVl.2, NaVl.3,
NaVl.4, NaVl.5, NaV1.6, NaVl.?, NaV1.8, NaV1.9, or CaV2.2 is implicated in the disease,
condition, or disorder. When activation or hyperactivity of NaVl.I, NaVl.2, NaVl.3, NaV1.4,
NaV1.5, NaVl.6, NaVl.7, NaVl.8, NaVl.9, or CaV2.2, is implicated in a particular disease,
condition, or disorder, the disease, condition, or disorder may also be referred to as a "NaVl.I,
NaV1.2, NaV1.3, NaVl.4, NaV1.5, NaV1.6, NaV1.?, NaV1.8, NaV1.9, or CaV2.2-mediated
disease, condition or disorder" or a "CaV2.2-mediated condition or disorder". Accordingly, in
another aspect, the present invention provides a method for treating or lessening the severity of a
disease, condition, or disorder where activation or hyperactivity of one or more of NaVl.l,
NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV1.?, NaV1.8, NaVl.9, or CaV2.2 is implicated
in the disease state.
[00119]
The activity of a compound utilized in this invention as au inhibitor of NaVl.l,
NaV1.2, NaV1.3, NaVl.4, NaVl.5, NaVl.6, NaVl.?, NaV1.8, NaV1.9, or CaV2.2 may be
assayed according to methods described generally in the Examples herein, or according to
methods available to one of ordinary skill in the 8.1.1.
[00120]
In certain exemplary embodiments, compounds of the invention are useful as
.inhiPit~rs of NaVI. 8.. In other ~bQ(limentS, compomids of the invention are useful as inhibitors
'... ; .... Qf·NaVi".~ and:Cll-YZ.2.. ·m ~other
,"
'.
-
.
'.
em6odhnei1~,
.
'.'
c9mpo~~ of the.....
in~~nij'oii ar~.~sefw~·
".
.." ..
"
inhibitors of CaV2.2.
[00121]
It will also be appreciated that certain of the compounds of present invention can exist
in free form for treatment, or where appropriate, as a pharmaceutically acceptable derivative
thereof. According to the present invention, a pharmaceutically acceptable derivative includes,
. but is not limited to, .pharmaceutically acceptable salts, esters, salts of such esters, ·or any other
adduct or derivative which upon administration to a patient in need is capable of providing,
directly or indirectly, a compound as otherwise described herein, or a metabolite or residue
thereof.
[00122J
As used herein, the term "pharmaceutically acceptable salt" refers to those salts which
are, within the scope of sound medical judgement, suitable for use in contact with the tissues of
-211-
WO 2005/042497
PCTlUS2004/036297
humar~ and lower animals without undue toxicity, irritation, allergic response and the like, and
are commensurate
means
with a reasonable
any non-toxic
administration
benefit/risk
ratio. A "pharmaceutically
salt or salt of an ester of a compound
of this invention
to a recipient, is capable of providing, either directly or indirectly,
this invention or an inhibitorily active metabolite
"inhibitorily
acceptable
or residue thereof.
salt"
that, upon
a compound of
As used herein, the term
active metabolite or residue thereof" means that a metabolite
or residue thereof is
also an inhibitor of a voltage-gated sodium ion channel.
[00123]
Pharmaceutically
Berge, et al. describe pharmaceutically
1977, 66, 1-19, incorporated
compounds
S. M.
acceptable salts in detail in J. Pharmaceutical Sciences,
herein by reference.
Pharmaceutically
acceptable
salts of the
of this invention include those derived from suitable inorganic and organic acids and
bases.
Examples
amino
group
phosphoric
salts are well known in the art. For example,
acceptable
of pharmaceutically
formed
acceptable,
with inorganic
acids
nontoxic acid addition
such as hydrochloric
acid, sulfuric acid and perchloric
salts are salts of an
acid,
hydrobromic
acid,
acid or with organic acids such as acetic acid,
oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other
methods used in the art such as ion exchange. Other pharmaceutically
adipate,
alginate, ascorbate, aspartate, benzenesulfonate,
camphorate,
camphorsulfonate,
ethanesulfonate,
.heptanoate,
citrate,
cyclopentanepropionate,
formate, fumarate, glucoheptonate,
hexanoate,
hydroiodide,
benzoate,
salts include
bisulfate,
borate, butyrate,
digluconate,
dodecylsulfate,
glycerophosphate,
2-hydroxy~ethanesulfonate;
acceptable
gluconate,
lactobionate,
hernisulfate,
lactate,. iaurate,'
la9i)rl s~~ie, .ci~att;,..P1al~~~~,
'inal(jn~te3 ~t;th~e~ilio~at~~.7~n;q>h~al~nes1,1l:fQn~t~,
~9otin~te,
'ci~~e,
~l~te: ox;Uate, palrnit~te,' p~~at~~
pecti~ate, pe~sulfate, 3-phe~ylpr~~ionate:
picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate,
undecanoate,
valerate salts, and the like. Salts derived from appropriate
metal, alkaline earth metal, ammonium
the quaternization
Water
or oil-soluble
Representative
magnesium,
or dispersable
p-toluenesulfonate,
bases include alkali
and W(Cl-4alkyl)4 salts. This invention
of any basic nitrogen-containing
products
groups of the coInpounds
may be obtained
by
such
also envisions
disclosed herein.
quatemization.
alkali or alkaline earth metal salts include sodium, lithium, potassium,
and the like. Further pharmaceutically
nontoxic ammonium, quaternary ammonium,
ph~sph~t~,
calcium,
acceptable salts include, when appropriate,
and amine cations fonned using counterions such
- 212-
WO 2005/042497
PCTlUS2004/036297
as hal~e, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl
sulfonate.
Pharmaceutically
[00124]
acceptable compositions
As described above, the pharmaceutically acceptable compositions of the present
invention additionally comprise a pharmaceutically acceptable carrier, adjuvant, or vehicle,
which, as used herein, includes any and all solvents, diluents, or other liquid vehicle, dispersion
or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents,
preservatives, solid binders, lubricants and the like, as suited to the particular dosage form
desired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack
'Publishing Co., Easton, Pa.,
1980) discloses
various
carriers
used in fommlating
pharmaceutically acceptable compositions and known techniques for the preparation thereof.
Except insofar as any conventional carrier medium is incompatible with the compounds of the
invention, such as by producing any undesirable biological effect or otherwise interacting in a
deleterious manner with any other component(s)
of the pharmaceutically
acceptable
composition, its use is contemplated to be within the scope of this invention. Some examples of
materials which can serve as pharmaceutically acceptable carriers include, but are not limited to,
ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum
albumin, buffer substances such as phosphates, glycine, sorbic acid, or potassium sorbate, partial
glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as
.pro~e.
sulf~l.te, disodium hydrogen' phosphate; potassium, hyc}rogeQphosphate, 'sodium
,c~op<;Je"~inc saItS,,coll~idal;sil!ca~'~gQ~~~
~si~pa~e,:ppl~nyl
'pyriy~d~n~, .po.lyacrylat~,~·
w:U:es,potyethyl~~e-polyo~ypr~pyl~e-block polymers, ~o~l fat: s~g~s s~~h a'~lact6~e,:gi~cose
and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as
sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth;
malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil,
cottonseed oil; safflower oil; sesame oil; olive oil; com oil· and soybean oil; .glycols; .such a
propylene glycol or polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar;
buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogenfree water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffer solutions, as
well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and
- 213-
PCTIUS2004/036297
WO 2005/042497
~
perfuming agents, preservatives and antioxidants can also be present in the composition,
according to the judgment of the formulator.
Uses of Compounds and Pharmaceutically Acceptable Compositions
[00125]
In yet another aspect, a method for the treatment or lessening the severity of acute,
. chronic, neuropathic, or inflammatory pain, arthritis, migrane, cluster headaches, trigeminal
neuralgia,
herpetic
neuralgia,
general
neuralgias,
epilepsy
or
epilepsy
conditions,
neurodegenerative disorders, psychiatric disorders such as anxiety and depression, myotonia,
arrythmia, movement disorders, neuroendocrine disorders, ataxia, multiple sclerosis, irritable
bowel syndrome, or incontinence is provided comprising administering an effective amount of a
compound, or a pharmaceutically acceptable composition comprising a compound to a subject in
need thereof. In certain preferred embodiments, a method for the treatment or lessening the
severity of acute, chronic, neuropathic, or inflammatory pain is provided comprising
administering an effective amount of a compound or a pharmaceutically acceptable composition
to a subject in need thereof. In certain embodiments of the present invention an "effective
amount" of the compound or pharmaceutically acceptable composition is that amount effective
for treating or lessening the severity of one or more of acute, chronic, neuropathic, or
inflammatory pain, epilepsy or epilepsy conditions, neurodegenerative disorders, psychiatric
disorders such as anxiety and depression, myotonia,
.
.
arrythmia, movement disorders,
.neuroendocrine dis,orders,at<pU.ll;
multiple ·sclerosis,iIritabie bowel syticJ.i.o:roe.,
.orincontinence.
tqo~6]:,,:.
may
~e
~oJ?lptilinds~~ c~mposition's, aC9P~g to ·th~.~e;thod '~fQ1e:p~e.1l:t.mventlo~,
be ~dmi~st~red' ri~ing'any amount and' any' route of ~dnrini~tration ~ffe~ti~~fo~tre~ting or '.'
lessening the severity of one or more of acute, chronic, neuropathic, or inflammatory pain,
epilepsy or epilepsy conditions, neurodegenerative disorders, psychiatric disorders such as
anxiety and depression, myotonia, arrythmia, movement disorders, neuroendocrine disorders,
ataxia, multiple sclerosis,.irritable bowel syndrome, or incontinence. The exact amount required
will vary from subject to subject, depending on the species, age, and general condition of the
subject.,the severity of the infection, the particular agent, its mode of administration, and the like.
The compounds of the invention are preferably formulated in dosage unit form for ease of
administration and uniformity of dosage. The expression "dosage unit form" as used herein refers
to a physically discrete unit of agent appropriate for the patient to be treated. It will be
·214·
PCT/US2004/036297
WO 2005/042497
>-
understood,
however,
that the total daily usage of the compounds
and compositions
of the
present invention will be decided by the attending physician within the scope of sound medical
judgment.
The specific effective dose level for any particular patient or organism will depend
upon a variety of factors including the disorder being treated and the severity of the disorder; the
activity of the specific compound employed; the specific composition employed; the age, body
weight,
general health,
administration,
sex and diet of the patient;
the time of administration,
and rate of excretion of the specific compound employed;
route of
the duration of the
treatment; drugs used in combination or coincidental with the specific compound employed, and
like factors well known in the medical arts. The term "patient", as used herein, means an animal,
preferably a mammal, and most preferably a human.
[00127]
The pharmaceutically
to humans
and other animals
intraperitone~y,
spray,
acceptable compositions
orally,
topically (as by powders,
or the like, depending
embodiments,
rectally,
the compounds
of this invention can be administered
parenterally,
ointments,
on the severity
intracisternally,
intravaginally,
or drops), bucally, as an oral or nasal
of the infection
being
of the invention may be administered
In certain
treated.
orally or parenterally
dosage levels of about 0.01 mglkg to about 50 mg/kg and preferably
at
from about 1 mg/kg to
about 25 mg/kg, of subject body weight per day, one or more times a day, to obtain the desired
therapeutic effect.
[00128]
Liquid
" pnarmaGeutjcally
dosage
forms
acc~ptable
for oral administration
emulsions,
include,
riJicroem.ulsions', solutions;
but
are not limited
suspensions,
, elixiis.:In .~dditi?n to ,the..~ctiYe.co~C?u1?-ds.,the liquj~ dQsagf?J6i:ms 'piay pori~
co~only
u~ed in
th~~
~~ch ~, 'ior ex~pl~"
emUlsifiers such as ethyl alcohol, isopropyl
alcohol, benzyl benzoate, propylene
particular,
cottonseed,
tetrahydrofurfuryl
groundnut,
s"rops. and
in~rt; 4ilpents, :" :.';,
~ater ~r other' ~olv~~ts, s~lubilizin~ '~ge~~~
alcohol, ethyl carbonate,
alcohol, polyethylene
germ,
olive,
castor,
and
~d .
ethyl acetate, benzyl
glycol, 1,3-butylene glycol, dimethylformamide,
com,
to,
sesame
oils),
oils (in
glycerol,
glycols and fatty acid esters of sorbitan~ and mixtures
thereof. Besides inert diluents, the oral compositions
can also include adjuvants such as wetting
agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
[00129]
Injectable
preparations,
for example,
sterile
injectable
aqueous
or oleaginous
suspensions may be formulated according to the known art using suitable dispersing or wetting
agents and suspending agents. The sterile injectable preparation may also be a sterile injectable
- 215·
WO 20051042497
PCTlUS2004/036297
)4-
solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for
example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may
be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In
addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For
this purpose any bland fixed oil can be employed including synthetic mono- or ~igJycerides. In
addition, fatty acids such as oleic acid are used in the preparation of injectables.
[00130]
The injectable formulations can be sterilized, for example, by filtration through a
bacterial-retaining filter, or by incorporating sterilizing agents in the fonn of sterile solid
compositions which can be dissolved or dispersed in sterile water or other sterile injectable
medium prior to use.
[00131]
In order to prolong the effect of a compound of the present invention, it is often
desirable to slow the absorption of the compound from subcutaneous or intramuscular injection.
This may be accomplished by the use of a liquid suspension of crystalline or amorphous material
with poor water solubility. The rate of absorption of the compound then depends upon its rate of
dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively,
delayed absorption of a parenterally administered compound form is accomplished by dissolving
or suspending the compound in an oil vehicle. Injectable depot forms are made by forming
microencapsule matrices of the compound in biodegradable polymers such as polylactidepolygl~colide. Depending upon the ratio of compound to po~ymerand the nature of the particular
.'polymer employed, the.. rate .'or"compound release .can be ·controlled. ExampleS of· other
" :.,·1?iocleg(ar1abl~"p~lyin~
•
'..
.:"
"0
incl~de polY(Qruio.es~ersY·
~d. po1Y(anhy'dIi~$).. Depot ;injeCt~ble
-
•
'.
"
."
"0 " ••
·0..
•
.'
•
...
.0'
•
"
•••••
1"
• ' ...
_ '''',
formulations are also prepared by entrapping the compound in liposomes or microemulsions that .
are compatible with body tissues.
[00132]
Compositions for rectal or vaginal administration are preferably suppositories which
can be prepared by mixing the compounds of this invention with suitable non-irritating
excipients or carriers such as cocoa butter, polyethylene glycol Qr a suppository wax which are .
solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or
vaginal cavity and release the active compound.
[00133]
Solid dosage forms for oral administration include capSUles, tablets, pills, powders,
and granules. In such solid dosage forms, the active compound is mixed with at least one inert,
pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate
- 216-
PCTlUS2004/036297
WO 2005/042497
and/ota) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic
acid,
b)
binders
such
as, for example,
carboxymethylcellulose,
alginates,
gelatin,
polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating
agents such as agar--agar, calcium carbonate, potato or tapioca starch, alginic acid, certain
silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption
accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example,
cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i)
lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium
lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form
may also comprise buffering agents.
Solid compositions of a similar type may also be employed
[00134]
as fillers in soft and hard-
filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular
weight polyethylene glycols and the like. The solid dosage fonDS of tablets, dragees, capsules,
pills, and granules can be prepared with coatings and shells such as enteric coatings and other
coatings well known in the pharmaceutical formulating art. They may optionally contain
opacifying agents and can also be of a composition that they release the active ingredient(s) only,
or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
Examples of embedding compositions that can be used include polymeric substances and waxes.
Solid compositions of a similar type may also be employed as fillers in soft and hard-filled
gelatin
~psUles u~in,gsuch
oq:nilk sugar as well
weight.
.
. .
. . -. eX~ipiehts.as la.ctose
.'
. as .high.molecular
.
. polethylene glycols ~d the like. '.'...., ...':" '.
.' .
. . .'
".
.,
'.'
•
....
."
"[00135]
I
•
.'
•.••
'!'
'.
.......
','
.•'.
.
.'
',....
.:.,
. ~..
.
',"
.•.• :. ."" .
.".
__ •
-
......
. The' active coinpounds can also be 'in microencapsulated form"with one 'or more
excipients as noted above. The solid dosage forms of tablets, dragees, capsules, pills, and
granules can be prepared with coatings and shells such as enteric coatings, release controlling
coatings and other coatings well known in the phannaceutical formulating art. In such solid
dosage forms th~ acqve cO:Qlpoundmay b~ adpllxed with at leas~ one..inert diluent such as
sucrose, lactose or starch. Such dosage forms may also comprise, as is normal practice,
additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids
such a magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and
pills, the dosage forms may also comprise buffering agents. They may optionally contain
opacifying agents and can also be of a composition that they release the active ingredient(s) only,
- 217-
"l
PCTmS2004/036297
WO 2005/042497
or prJ'erentially, in a certain part of the intestinal tract, optionally, in a delayed manner.
Examples of embedding compositions that can be used include polymeric substances and waxes.
[00136]
Dosage fonus for topical or transdermal administration of a compound of this
invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants
or patches. The active component is admixed under sterile conditions with a pharmaceutically
acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic
formulation, eardrops, and eye drops are also contemplated as being within the scope of this
invention. Additionally, the present invention contemplates the use of tninsdermal patches,
which have the added advantage of providing controlled delivery of a compound to the body.
Such dosage forms are prepared by dissolving or dispensing the compound in the proper
.
'
medium. Absorption enhancers can also be used to increase the flux of the compound across the
skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing
the compound in a polymer matrix or gel.
[00137]
As described generally above, the compounds of the invention are useful as inhibitors
of voltage-gated sodium ion channels. In one embodiment, the compounds and compositions of
the invention are inhibitors of one or more of NaVl.l, N.aV1.2, NaV1.3, NaVl.4, NaV1.5,
NaV1.6, NaVl.?, NaV1.8, NaVl.9, or CaV2.2 and thus, without wishing to be bound by any
particular theory, the compounds and compositions are particularly useful for treating or
lessening the severity of a disease, condition, or disorder where activation or hyperactivity of
one or:inore of NaVl.l, NaV1.2, NaV1.3, NaVlA',.NaVli5, N~VI:6, NaV'1~7,NaV1.8, NaVi.9,·
Qt. ¢aV7.2
ot.
i~ j,inpUc~ted,ip·th~d;isease?,?on,dition, dis~~
•.."~en
or hYPera~tivitY..
a~~vatiQri
of·NaV1.1, N~V·1.2·:NaV1.3: N~VI.·4, NaVl~5, NaV1.6, NaVl~7,"N~V1.8·,N~V1.9, or C~V2.2
is implicated in a particular disease, condition, or disorder, the disease, condition, or disorder
may also be referred to as a "NaV1.1, NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV1.?,
NaV1.8, NaV1.9, or CaV2.2-mediated disease, condition or disorder". Accordingly, in another
aspect, the present invention provides a method for treating or lessening the severity 'Ofa disease,
condition, or disorder where activation or hyperactivity of one or more of NaV1.1, NaV1.2,
NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaVl.?, NaV1.8 or NaV1.9 is implicated in the disease
state.
- 218-
..
WO 2005/042497
[00138]
NaV1.2,
PCTfUS2004/036297
The activity of a compound
utilized in this invention as an inhibitor of NaV!.1,
NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV1.7, NaV1.8, NaV1.9,
or CaV2.2 may be
assayed according to methods described generally in the Examples herein.
It will also be appreciated
[00139]
compositions
that the compounds
and pharmaceutically
of the present invention can be employed in combination
compounds
and pharmaceutically
acceptable
compositions
therapies, that is, the
can be administered
with, prior to, or subsequent to, one or more other desired therapeutics
acceptable
concurrently
or medical procedures.
The· particular combination of therapies (therapeutics or procedures) to employ in a combination
regimen will take into account compatibility
effect to be achieved. It will also be appreciated
the desired therapeutic
employed
of the desired therapeutics andlor procedures and
may achieve
a desired effect for the same disorder
that the therapies
(for example,
an inventive
compound may be administered concurrently with another agent used to treat the same disorder),
or they may achieve different effects (e.g., control of any adverse effects). As used herein,
additional
therapeutic
agents that are normally
administered
to treat or prevent a particular
disease, or condition, are known as "appropriate for the disease, or condition, being treated".
[00140]
The amount of additional
therapeutic
agent present in the compositions
of this
invention will be no more than the amount that would normally be administered in a composition
that therapeutic agent as the only active agent. Preferably the amount of additional
comprising
therapeutic agent in the presently disclosed compositions
the ~ount
noCmanY pi'esent)n.a
'.. ~cti.ve i!-gent.
'.'
·ne·
. [~i)141]'....
.....
.. ...
.,
.. ,
of this inv~ntion' or phaImacerit~citIlY ·accept~bie· compositions
thereof may also be incorporated
such as prostheses,
compQsition comprising that agept as the.onlY therapeutiG~ly
. .....
'c~~pounci~
will range from about 50% to 100% of
into compositions
for coating an implantable medical device,
artificial valves, vascular grafts, stents and catheters.
present invention, in another aspect, includes a composition
Accordingly,
for coating an implantable
the
device
.coIQ.prising a compound of the present invention as described generally above, and in classes and
subclasses
aspect,
herein, and a carrier suitable for coating said implantable
the present
invention
includes
an implantable
device
device.
In still another
coated with a composition
comprising a compound of the present invention as described generally above, and in classes and
subclasses herein, and a carrier suitable for coating said implantable
device.
Suitable coatings
and the general preparation of coated implantable devices are described in US Patents 6,099,562;
- 219-
peT IUS2004/036297
wo 2005/042497
~
5,886,026; and 5,304,121.
The coatings are typically biocompatible polymeric materials such as
a hydrogel polymer, polymethyldisiloxane,
polycaprolactone,
acid, ethylene vinyl acetate, and mixtures thereof.
covered
by
phospholipids
a suitable
topcoat
or combinations
of
fluorosilicone,
thereof
to impart
polyethylene
glycol, polylactic
The coatings may optionally be further
polys accari des,
controlled
release
polyethylene
glycol,
characteristics
in the
composition.
[00142]
Another
aspect of the invention
relates to inhibiting
NaVl.1,
NaV1.2, NaV1.3,
NaV1.4, NaV1.5, NaV1.6, NaV1.7, NaV1.8, NaV1.9, or CaV2.2 activity in a biological sample
or a patient, which method comprises administering to the patient, or contacting said biological
.sample with a .compound of formula I or a composition comprising said compound.
The term
.''biological sample", as used herein, includes, without limitation, cell cultures or extracts thereof;
biopsied material obtained from a mammal or extracts thereof; and blood, saliva, urine, feces,
semen, tears, or other body fluids or extracts thereof.
[00143]
Inhibition of NaVl.l,
NaV1.2, NaV1.3, NaV1.4, NaV1.5, NaV1.6, NaV1.?, NaV1.8,
NaV1.9, or CaV2.2 activity in a biological sample is useful for a variety of purposes that are
known to one of skill in the art. Examples of such purposes include, but are not limited to, the
study of sodium ion channels in biological and pathological
phenomena;
and the comparative
evaluation of new sodium ion channel inhibitors.
[00144]
In order that the invention described herein may be more fully understood,
. :following"e},Camplesare.s~t.fqrtP:. ~t shol,lld be'qnderstood'that
'
. ..".
p~ose~
these examples are for illustrative
only a~.lf~~. DC!tto~e:c<?il~tnie~~ limitlQg tlus-inyentiq!1 in.ani~anner.
..'
......:..
'.'
..
'
.... .....
: ~.'
EXAMPLES
SYNTHESIS OF EXEMPLARY COMPOUNDS OF THE INVENTION:
EXalIl,ple 1:
- 220-
the
.
WO 2005/042497
PCTIUS2004/036297
~
This example teaches the preparation of [2-(1-methyl-1H-benzoimidazol-2-yl)-ethyl]-carbamic
acid tert-butyl ester (6). To a solution of beta-alanine (3.5 g, 18.5 mmol) and pyridine (3 rnL,37
mmol) in dichloromethane
(20 mL) was added PFP ester (3.2 mL, 18.5 mmol).
room temperature for 1 hour, 1,2-phenyleneamine
mL) was added to the reaction mixture.
water, extracted with dichloromethane
After stirring at
1 (2 g, 18.5 nunol) in dichloromethane
(20
After stirring overnight, the reaction was quenched with
(150 mL*2), dried and removed the solvent. Then the
crude mixture was triturated with dichloromethane
(20 mL), filtered to give white solid 2 as the
desired product (4.8 g) at 90% yield. LCIMS (10-99%) M+lIZ 280.3 retention time 2.03 min.
H
ero
~~NH80C.
cr~~NHBOC
NH2
3
2
[00145]
Compound 2 (4 g) was dissolved in acetic acid (50 mL) and heated to 65°C for 1 h,
then cooled down to room temperature,
with dichloromethane,
removed the solvent in vacuo. The residue was taken up
quenched with sat NaHC03, extracted with diChloromethane
(150 mL)
(pH = 10), concentrated to afford 3 as light yellow solid (3.7 g) at 100% yield. LCIMS (10-99%)
M+ 1/Z 262.2 retention time 2.30 min.
r(Y~
'.'
.~~
. ~~~NHB~C
:.
'"0'
'.
.',--:--~--.....
.
'. ~",
~
I',
••.•
•
H~I
.
~~~NH2.~OI'
• ':.•
'
.'
• '.
.
'.
'..
..•••
3
[00146]
•
4
Compound 3 (3 g) was dissolved in 3 N HClIEtOAC
room temperature
"','
0""
overnight.
compound 4 as pink solid
(50 mJ.125 mL) and stirred at
The solvent was in vacuo and dried in high vacuum to give
(2.4 g) at greater than 95% yield.
LCIMS (10-99%) M+lIZ
retention time 0.62 inin.
5
4
- 221-
162.4
WO 2005/042497
[001471
PCTIUS2004/036297
To a solution of benzimidazole
mmol), 3,4-dimethylphenoxyacetic
ethylene amine dihydrochloride
acid (77 mg, 0.43 mmol) , and BOP reagent (190 mg, 0.43
(5 mL) was added triethylamine (0.23 mL, 1.72 mmol).
nunol) in acetonitrile
(100 mg, 0.43
room temperature overnight, the solvent was removed under vacuum.
After stiffing at
The residue was takt:n up
with water and saturated sodium bicarbonate (20 mL), extracted with dichloromethane
2), dried and removed the solvent.
(30 mL x
The crude mixture was purified by using Gilson HPLC to
give white solid 5 as a TFA salt (170 mg) at 93 %yield. MUX LCIMS (10-99%) M+1/Z 324.163
retention time 2.62 min; lH NMR (DMSO) <52.12 (s, 3R), <52.15 (5, 3H), <52.99 (t, 2H, J
Hz), B 3.59 (m, 2H), (54.40 (s, 2H), (56.64 (dd, IH, J = 2 Hz, 6.5 Hz), 0 6.75 (d, IH, J
= 5.88
= 1.8 Hz),
86.98 (8, 1H, J = 6.6 Hz), 8 7.12 (m, 2H), 7.40 (d, 1H, J = 5.76 Hz), 7.53 (m, HI), 8.30 (t, 1H, J
= 4.56 Hz), 12.24 (s, IH); 13C NMR (DMSO) 0 18.4,
B 111.58,
8143.24,
<5 116.13,
S 152.74,
B 118.12,
<5 155.73,
<5 120.86,
.5 121.52,
<5 19.57,
S 28.58, <5 36.74, 067.07,
<5 128.70,
S 130.12, .5 134.21,
<5 110.8,
<5 137.25,
S 167.85.
Me
H
~N»-~:
O:~~NHBOC
6
3
[00148]
"'-.-NHBoc
Boc-protected
benzimidazole
ethyl amine 3 (400 mg, 1.53 rnmol) was dissolved in
THF (50 mL) and cooled to DoC under nitrogen, followed by dropwise addition of LiHMDS (535
. mg,
3.37 mmol)
stirred fQ~ iO ..~.
~x·~e.··
in t.ffp(10
mL)
via sYringe.'
Me~hyl.iPdide
Aft~~"stici~g
'f~;5'h
The mlxtUre was waimed
'0:).1.~, .1.68. ~Qlt
:o/as~:added
'to roo~ temperatUre and ..
dropwi~e to ,the reactjo~, .'
at r~o~ 't~mperature, 'the 're~cti~n 'wa~ quen~hed ~ith '';;;t~r (20
mL) and extracted with EtOAc (30 mL x 2), dried, concentrated
and purified by ISCO flash
chromatography.
Example 2:
o
Ji'0H
This example teaches the preparation
dimethyl-benzyl
ester.
. ~J-CI
of [2-(IH-benzoimidazol-2-yl)-ethyl]-carbamic
To a solution of 3,4-dimethylbenzyl
- 222-
alcohol (1.5 g, 11.0
acid 3,4mmol) in
.
peT IUS2004f036297
WO 2005/042497
.,;
toluene (20 mL) was added phosgene (13 roL, 24.3 mmol, 20% in toluene). After stirring at
room temperature overnight, excess phosgene and toluene was removed in vacuo, and dried
under high vacuum for 2 h. providing 3,4-dimethylbenzylchloroformate (1.8 g) as an oil at 80 %
yield. (reference: Nagele, E.; Schelhaas, M.; Kuder, N.; Waldmann, H. 1. Am. Chem. Soc. 1998,
120,6889)
Example 3:
H
cr~~NH
..
}-N~
o
y
CI
CI
This example teaches the preparation of 1-[2-(lH-benzoimidazol-2-yl)-ethyl]-3-(3,4-dichlorobenzyl)-urea. To a solution of benzimidazole ethyl amine dihydrochloride (50 mg, 0.21 mmol)
in pyridine (1.5 mL) was added 3,4-dichlorobenzylisocyanate (43 p.L, 0.21 mmol). After stirring
overnight at room temperature, the crude mixture was purified by Gilson HPLC to afford white
solid (50 mg) as a TFA salt at 92% yield. LCIMS (10-99%) M+l/Z 324.2 retention time 2.97
min.
Example 4:
H
H
~N
0
•'.. ··~0N;S~
O=~hy",
~.
0
.• " " .. , •. '. '" ,.'
• " :-<::1·
..
.'.
'..
This example teaches the preparation of N-[2-(lH-Benzoimidazol-2-yl)-ethyl]-3,4-dichlorobenzenesulfonamide. To a solution of benzimidazole ethyl amine dihydrochloride (50 mg, 0.21
mmol) in pyridine (1.5 mL) was added 3,4-dichlorobenzyisocyanate (58 p.L, 0.21 mmol). After
stirring overnight at room temperature, the crude mixture was purified by Gilson HPLC to afford
. white solid (55 mg) as TFA salt at 92%·yield.
Example 5:
F
+
6:
I
~
HO
F C
-223 -
WO 2005/042497
PCTlUS2004/036297
j'
This example teaches the preparation of 2-(2,6-Difluoro-phenyl)-7-methyl-lH-benzoimidazole
according to the following procedure.
A solution of 3-methyl-benzene-l,2-diamine
in EtOH (2 mL) was heated for 5 min at 180°C in a
0.82 mmol) and 1,6-difluorobenzaldehyde
microwave synthesizer.
(100 mg,
The EtOH was removed, the residue was dissolved in DMSO (l rnL),
and was purified on a Gilson HPLC to afford 2-(2,6-difluoro-phenyl)-7-methyl-1Hbenzoimidazole
as 1FA salt (200 mg) at 100% yield. MUX LCIMS (10-99%) M+l/Z 245.061
retention time 2.1 min.
[00149]
Other compounds of Formula I have been prepared by methods substantially similar
to those described above. The characterization
data for these compounds is summarized in Table
3 below. The compound numbers correspond to the compound numbers listed in Table 2.
Table:3
Characterization Data for Selected Comoounds of Fomula I from Table 2
LCIMS
LCIRT
LCIRT
LCIMS
Cmpd#
Cmpd#
(min)
(min)
M+
M+
3
14
16
23
49
54
66
165
1'71
189
·192
196
197
1.9~,
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
256.28
336.40
326,20
324.40
326.20
344.00
372.20
324.20
364.00
324.20
324.40
. 35820
332.00'
.. 321.20'
309.20
314.00
313.20
329.20
309.20
363.20
364.00
365.20
379.20
364.00
339.20
372.20
372.20
386.00
372.20
346.20
338.20
3.20
2.63
1.90
3.74
2.75
2.94
2.46
1.84
2.32
1.69
2.79
2.45
2.87
...
.
''L8S' .
2.06
2.16
1.91
1.97
1.96
2.17
2.25
2.19
2.35
2.25
2.14
2.15
2.09
2.22
2.07
2.04
3.13
'"
.
'
:"
"
,
,
..
...
"."
"
,
,-.
..
' ,
-224-
266
267
268
269
270
271
272
273
274
275
276
277
'278 '
,'219:'
280
281
282
283
284
285
286
. '281
288
289
290
291
292
293
294
295
296
344.20
323.20
370,00
336.40
344,00
346.00
340.00
324.00
325.20
366.82
.. 34MO
315.78
267.30
". '346.85.
335.30
326.43
284.33
323.41
317.36
321.20
339.20
354.20
353.20
366.20
350.20
382.00
372.00
372.00
332.00
338.20
348.00
3.00
2.89
2.89
2.08
3.90
3.70
3.63
3.81
3.18
3.20
3.30
3.10
3.30
. .3~30
3.10
3.50
3.50
3.10
3.10
1.88
1.76
2;28
1.55
2.16
2.06
2.96
2.84
3.07
2.75
2.62
2.90
..
..
.
',:.
','
PCTlUS2004/036297
WO 2005/042497
Cmpd#
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
.. 327
.' 328'
..'. 329
: "330
331
332
333
LC/MS
M+
373.00
384.00
400.20
384.00
413.00
358.00
356.20
400.20
352.20
398.20
380.00
320.20
398.20
321.20
332.20
338.20
380.00
388.20
386.20
422.00
381.00
386.20
388.20
381.40
388.20
386.20
408,20
328.20
401.20
372.20
400.20
'398.00
347.20
. : '338.20 .
347.20
381.40
353.20
LCIRT
Cmpd#
(min)
2.66
2.85
3.00
2.77
3.10
3.09
2.96
2.29
1.68
2.44
2.35
2.11
1.88
1.90
2.01
1.90
2.37
2.49
2.52
2.46
2.01
2.57
2.23
1.68
2.34
2.69
2.34
2.15
2.10
2.42
2.26
2.43
1-.'30' .
,
1$1:
1.26
1.61
1.74
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
366
367
368 '
" ',369'"
370
LCIMS
M+
LCIRT
(min)
335.40
1.63
335.20
1.86
383.20
1.73
374.20
1.65
339.00
1.46
408.40
1.44
347.20
1.44
381.40
1.53
415.20
2.15
364.40
1.79
388.20
2.39
362.20
2.27
342.20
2.26
2.05
421.00
350.20
2.13
2.31
376.00
2.23
382.00
2.46
404.40
2.37
376.00
406.20
2.26
404.20
2.63
2.06
328.00
342.00
1.82
344.00
1.87
350.20
2.02
350.20
1.89
350.20
1.98
368.00
1.96
366.00
1.99
2.08
350.20
2.54
.334.00
' 2.75
350:20
. '316.20
·2.45'
"'3,30.20' .: . ' '2:t';iS:'.
344.00
2.83
Micromass MUX LCT 4 channel LCIMS, Waters 60F pump, Gilson 215 4 probe
autosampler, Gilson 849 injection module, 1.5 mL/minlcolumn flow rate, 10-99% CH3CN
'(0.035'% TFA) IHzO (0.05 % 'ITA) gradient, PhenomenexLuna 5u CIS columns (50 X'
4.60 nun), Waters MUX UV·2488 UV detector, Cedex 75 ELSD detectors.
ASSAYS FOR DETECTING AND MEASURING NaV INHIBITION PROPERTIES OF
COMPOUNDS
[00150]
A) Optical methods for assaying NaV inhibition properties of compounds:
-225 -
PCTIUS2004/036297
WO 2005/042497
-'I
[00151]
Compounds
of the invention are useful as antagonists
channels.
Antagonist properties of test compounds were assessed as follows,
the Na V of interest were placed into'microtiter
stained with fluorescent
of voltage-gated
sodium ion
Cells expressing
plates. After an incubation period, the cells were
dyes sensitive to the transmembrane
potential.
The test compounds
were added to the microtiter plate. The cells were stimulated with either a chemical or electrical
means to evoke a NaV dependent membrane potential change from unblocked channels, which
was detected
and measured with trans-membrane
potential-sensitive
dyes.
detected as a decreased membrane potential response to the stimulus.
potential
assay utilized voltage-sensitive
Antagonists
The optical membrane
FRET sensors described by Gonzalez and Tsien (See,
J. E. and R. Y. Tsien (1995) "Voltage sensing by fluorescence
Gonzalez,
were
resonance energy
transfer in single cells" Biophys J 69(4): 1272-80, and Gonzalez, J. E. and R. Y. Tsien (1997)
"Improved
transfer"
indicators
Chern
fluorescence
Bioi
of cell membrane
4(4): 269-77)
potential
that use fluorescence
in combination
changes such as the VoltagelIon
with
resonance
instrumentation
energy
for measuring
Probe Reader (VlPR®) (See, Gonzalez, J. E., K.
Oades, et al, (1999) "Cell-based assays and instrumentation
for screening ion-channel targets"
Drug Discov Today 4(9): 431-439).
[00152]
B) VIPR® optical membrane potential assay method with chemical stimulation
Cell Handling and Dye Loading
[00153]
24 hours before the assay on VIPR. CHO cells endogenously
type voitage-gated
Other subtypes'
.
NaV
~eseeded. in 96-well
ar~perfoP:ne,cl"in~,8naiogous
expressing a Na V1.2
poly-lysine coated piates at 60.000 cells per well:
v ~iln~res_t:".
,', :, ' .
....
.....
iQo,d(d~ a cell, fine"eXPreSSJ~g"the'Na
...
. .. ".
....
. . .
"
1) On the day of the assay, medium is aspirated and cells are washed twice with 225
J.IL of Bath
Solution #2 (BS#2).
2) A 15 uM CC2-DMPE
solution is prepared by mixing 5 mM coumarin
stock solution with
10% Pluronie 127 1:1 and then' dissolving the:mix in the appropriate volume of BS#2.
3) After bath solution is removed from the 96-well plates, the cells are loaded with 80 I!L of the
CC2-DMPE
solution. Plates are incubated in the dark for 30 minutes at room temperature.
4) While the cells are being stained with coumarin,
prepared. In addition to DiSBAC2(3),
I-IL veratridine
a 15
J.IL
oxonol
this solution should contain 0.75
(prepared from 10 mM EtOH stock. Sigma #V-5754).
- 226-
solution in BS#2 is
roM ABSCI and 30
PCT/US2004/036297
WO 2005/042497
~
5) After 30 minutes, CC2-DMPE
is removed and the cells are washed twice with 225
J.IL
of
BS#2. As before, the residual volume should be 40 ilL.
6) Upon removing the bath, the cells are loaded with 80 ilL of the DiSBAC2(3)
solution, after
which test compound, dissolved in DMSO, is added to achieve the desired test concentration
to each well from the drug addition plate and mixed thoroughly.
The volume in the well
should be roughly 121 ilL. The cells are then incubated for 20-30 minutes.
7) Once the incubation is complete, the cells are ready to be assayed on VIPR® with a sodium
addback protocol.
depolarization.
120 ilL of Bath solution #1 is added to stimulate the NaV dependent
J.IL tetracaine
200
was used as an antagonist positive control for block of the
NaV channel.
Analysis of VIPR
[00154]
®
Data:
Data are analyzed
and reported
as normalized
ratios
of background-subtracted
emission intensities measured in the 460 nm and 580 run channels.
then subtracted from each assay channel.
Background
intensities are
Background intensities are obtained by measuring the
emission intensities
during the same time periods from identically treated assay wells in which
there are no cells.
The response as a function of time is then reported as the ratios obtained
.usinK the fol1<?winRformula:
.
. (intensity.:460 ~ . ~'baCkgrom1d~o~)'
.
R(t) = ---------------------------------------(intensity
[00155]
580
nm - background
580
nm)
The data is further reduced by calculating the initial (Ri) and final (Rr) ratios.
are the '~v~rag~ ratio v;U~es duri~g part or all of th~ pre-;timulation
points during the stimulation
period.
The response to the stimulus
These
period, and during ~~ple
l? = RflRj
is then calculated.
For the Na+ addback analysis time windows, baseline is 2-7 see and final response is sampled at
15-24 sec.
[00156]
Control responses
with the desired
properties
are obtained by performing
(positive
control),
assays in the presence of a compound
such as tetracaine,
- 227-
and in the absence
of
PCTlUS2004/036297
WO 2005/042497
k
pharmacological agents (negative control).
Responses to the negative (N) and positive (P)
controls are calculated as above. The compound antagonist activity A is defined as:
A
= R - P * 100 .
N-P
where R is the ratio response of the test compound
Solutions [roM]
Bath Solution #1:
NaCI 160, KCI4.5, CaCl2 2, MgCl2 1, HEPES 10, pH 7.4 with NaOH
Bath Solution #2
TMA-CI160, eaCb 0.1, MgCh 1, HEPES 10, pH 7.4 with KOH (final K
concentration - 5 mM)
CC2-DMPE:
prepared as a 5 mM stock solution in DMSO and stored at -20°C
DiSBAC2(3):
prepared as a 12 mM stock in DMSO and stored at -20°C
ABSC1:
prepared as a 200 mM stock in distilled H20 and stored at room
temperature
Cell Culture
[00157]
CHO cells are grown in DMEM (Dulbecco's Modified Eagle Medium; GibcoBRL
#1O:S~9-o10).suppl~¢erit~4 wi~ 10% FBS (F~taIBovine· Seruin, 'qu~ifiedi Gibco;BRL'#1614.0:· '.
:(71) ~d. 1%.Pe~-S~p ·.(Peili¢illin,-'Sti.-eptomY~;:GibcoBRL#15~40':·122).. CeTIs..ate gtp-wn)p.'
'~erit~dca~'ti~s~, i~ 90% h~~dlt~'~;;
10'%'C~~, :to
100%'~onti~~n~~:'Th~Y ~e' tis~ally"~p~t'bY
trypsinization 1:10 or 1:20, depending on scheduling needs, and grown for 2-3 days before the
next split.
[00158J
C) VlPR® optical membrane potential assay method with electrical stimulation
.[00159J.
The following is an example of how NaVL3 inhibition activity is measured using the
optical membrane potential method#2. Other subtypes are performed in an analogous mode in a
cell line expressing the NaV of interest.
[00160J
HEK293 cells stably expressing NaV1.3 are plated into 96-well microtiter plates.
After an appropriate incubation period, the cells are stained with the voltage sensitive dyes CC2DMPE/DiSBAC2(3) as follows.
- 228-
~..
PCTlUS2004f036297
WO 2005/042497
Reagents:
100 mg/rnL PlUTonic F-127 (Sigma #P2443), in dry DMSO
10 mM DiSBAC2(3) (Aurora #00-100-010)
in dry DMSO
10 IruV1CC2-DNIPE
in dry DMSO
(Aurora #00-100-008)
200 roM ABSCI in H20
Hank's
Balanced
Salt Solution
(Hyc1one #SID0268.02)
supplemented
with 10 mM HEPES
(Gibco #15630-080)
Loading protocol:
= 20 JtM
[00161]
2X CC2-DMPE
equivalent
volume of 10% plUTOnic, followed
containing
10 mM HEPES.
CC2-DMPE
10 mM CC2-DMPE
by vortexing
is vortexed with an
amount of HBSS
in required
Each cell plate will require 5 mL of 2X CC2-DMPE.
is to wells containing
concentration.
washed
cells, resulting
in a 10
50 ILL of 2X
p.M final staining
The cells are stained for 30 minutes in the dark at RT.
2X DISBACz(3)
[00162]
CC2·DMPE:
with ABSC1
= 6JtM DISBACz(3) and 1 roM ABSC1:
The
required amount of 10 ruM DISBAC2(3) is added to a 50 ml conical tube and mixed with 1 ILL
10% plUTOniCfor each mL of solution to be made
and vortexed together.
Then HBSS/HEPES
is added to make up 2X solution. Finally, the ABSCI is added.
,[O01~3] 'Th~ ~X'])iSBAC2(3)
"
"
,-
. conipo~d,pl{ltes
.,
••
",._
•••
solutiqn can be',used to./>oIYl:ItecompoWId plates.
w:etrla9-e,at ~X'drqg co~cen~tiop..
' ~olu~~ 01"50 '~L:
'.....
•
Add
50 ill.:veli"of
•••
',...
Note
that
,W~sh staiJi¢ plate. again, le~;.mg:re:sjciti~·
"0.0
•
th~'2X'DiSBAC2(3)"w1
.'
ABSCi.
.,
to.
Stai~'f~~ '30'~~ute~
'hi"
the dark at RT.
[00164]
The electrical
stimulation
instrument
Channel Assay Methods PCTIUS01l21652,
comprises
a microtiter
simultaneously
plate handler,
and methods
herein incorporated
of use are described
by reference.
in ION
The instrument
an optical system for exciting. the coumarin dye ,while
recording the coumarin and axonal emissions, a waveform generator, a current-
or voltage-controlled
amplifier, and a device for inserting electrodes in well.
computer control, this instrument passes user-programmed
within the wells of the microtiter plate.
Reagents
- 229-
....
Under integrated
electrical stimulus protocols to cells
: , .',
.
PCTlUS2004/036297
WO 2005/042497
[0016St
Assay buffer #1
140 roM NaCI, 4.5 IIliVIKCI, 2 IIliYfCaCh, 1 mM MgCh. 10 mM HEPES,
10 roM glucose, pH
7.40, 330 mOsm
Pluronic stock (lOOOX): 100 mglmL pluronic 127 in dry DMSO
axonal
stock
(3333X): 10 roM DiSBAC2(3) in dry DMSO
(1000X): 10 roM CC2-D11PE in dry DMSO
Coumarin stock
ABSCI stock (400X): 200
roM ABSCI in water
Assay Protocol
1. rosert or use electrodes into each well to be assayed.
2. Use the current-controlled
amplifier to deliver stimulation
seconds of pre-stimulus recording are performed
wave pulses for 3 s. Two
to obtain the un-stimulated
intensities.
Five seconds of post-stimulation recording are performed to examine the relaxation to the
resting state.
Data Analysis
[00166]
Data
are analyzed
and reported
as normalized
ratios
emission intensities measured in the 460 nm and 580 nm channels.
then subtrac:t~
from eae}:!.assay chanhel.
4~i :the.~~e
e~ssiqn; m~n.sit!ei:;""
" " . ther~ ~e rio ·celi;.·
f:ime
Ba6kk!:oup.d.mtensfties
of background-subtracted
Background
intensities are
are' obtairi~(f by measuring toe'·
'1?~rio~~'rto:n.t: id~n~~~ly ·~at~d.~s~y. :wells in ~wNcli :'.
.The 'r~s~onse as a 'fun~ti~n of'timti~
the~
;~~~rt~d'~
th~rati~~~b~ain~ci
using the following formula:
(intensity
. ,R(t).:::;.
nm - background
460
nm)
----------.--------~7--.----"7--(intensity
[00167]
460
S80 nm -
background sso nm)
The data is further reduced by calculating the initial (Rj) and final CRf) ratios. These
are the average ratio values during part or all of the pre-stimulation
period, and during sample
points during the stimulation period. The response to the stimulus .R. ::: RtlRi is then calculated.
-230 '
.r.'
.
PCTIUS2004/036297
WO 2005/042497
[00168]
Control responses are obtained by perfonning assays in the presence of a compound
with the desired properties (positive control), such as tetracaine, and in the absence of
pharmacological agents (negative control). Responses to the negative (N) and positive (P)
controls are calculated as above. The compound antagonist activity A is defined as:
A= R-P *100.
where R is the ratio response of the test compound.
N-P
[00169]
ELEClROPHYSIOLOGY
ASSAYS FOR NaV ACTIVITY AND INHBITION OF
TEST COMPOUNDS
[00170]
Patch clamp electrophysiology was used to assess the efficacy and selectivity of
sodium channel blockers in dorsal root ganglion neurons. Rat neurons were isolated from the
dorsal root ganglions and maintained in culture for 2 to 10 days in the presence of NGF (50
nglmI) (culture media consisted of NeurobasalA supplemented with B27, glutamine and
antibiotics). Small diameter neurons (nociceptors, 8-12 J.Ul1 in diameter) have been visually
identified and probed with fine tip glass electrodes connected to an amplifier (Axon
Instruments). The "voltage clamp" mode has been used to assess the compound's IC50 holding
the cells at - 60 mV. In addition, the "current clamp" mode has been employed to test the
efficacy of the compounds in blocking action potential generation in response to current
injections. The results of these experiments have contributed to the definition of the efficacy
pro:file ~f the compoun~~ ,
. [00171]
.
,...-;.
VOLTAGE-CLAMP assay inDRG neurons' .
':"
[00172]
",
:'
'..
:'
.,"
',::
,
',~'
': '.'
:
," ,.. ,
,.:' ,": ,'".."
".
' ,:'
• ",
"
;' '" '
.. , ~"
..' :"',~_,
:'
l'"
ITX-resistant sodium CUITehts'wererecorded from DRGsomata using the whole-cell
variation of the patch clamp technique. Recordings were made at room temperature (-220 C)
with thick walled borosilicate glass electrodes (wpI; resistance 3-4 M.Q) using an Axopatch'
200B
amplifier
(Axon Instruments).
After establishing
the whole-cell
configuration,
'approximately 15 minutes' were' allowed for the pipette solution'to' equilibrate within the cell
before beginning recording. Currents were lowpass filtered between 2-5 kHz and digitally
sampled at 10 kHz. Series resistance was compensated 60-70% and was monitored continuously
throughout the experiment. The liquid junction potential (-7 mY) between the intracellular
pipette solution and the external recording solution was not accounted for in the data analysis.
- 231-
•
'.. ','
'
PCTIUS2004/036297
WO 2005/042497
~
Test solutions were applied to the cells with a gravity driven fast perfusion
system (SF-77;
Warner Instruments).
[00173]
Dose-response
relationships
were detennined
in voltage clamp mode by repeatedly
depolarizing the cell from the experiment specific holding potential to a test potential of +10mV
once every 60 seconds. Blocking effects were allowed to plateau before proceeding to the next
test concentration.
Solutions
[00174]
Intracellular
solution (in mM): Cs-F (130), NaCl (10), MgCIZ (1), BGTA (1.5),
CaC1z (0.1); HEPES (10), glucose (Z), pH = 7.4Z, Z90 mOsm.
[0~175]
Extracellular
solution (in mM): NaCI (138), CaCl2 (1.26), KCI (5.33), KHZP04
(0.44), MgCI2 (0.5), MgS04
(0,41), NaHC03
CdC12 (0.4 ), NiC12 (0.1), 1TX (0.25
(0.3), glucose (5.6), HEPES (10),
x 10-3).
[00176]
CURRENT-CLAMP
[00177]
Cells were current-clamped
amplifier
(4), NaZHP04
assay for NaV channel inhibition activity of compounds
(Axon Inst). Borosilicate
in whole-cell
configuration
with a Multiplamp
700A
pipettes (4-5 MOhm) were filled with (in mM):150 K-
gluconate, 10 NaCI, 0.1 EGTA, 10 Hepes, 2 MgCh, (buffered to pH 7.34 with KOB). Cells were
bathed in (in mM): 140 NaCI, 3 KCI, 1 MgCI , 1 CaC} , and 10 Hepes). Pipette potential was
~ero~d before seal formation;
,. Recprdi,ngs
. . .'
"
......
were made.~t ~oom te~pt;r~~
'
.:"
[00178]
liq~~ jpnction potentials weJe not corrected during acquisition.
ASSAYS
.. ,:
'.
FOR
,",
.
c'
..
. ',
. '.' ..
DETECTING
'.
;
.'
.•.. -.-
AND
.,_
.
..
.
:.
'.
'.',:'
. .. '.
•
.;.
.
.
'.:
,"
MEASURING
CaV
INHmITION
PROPERTIES OF COMPOUNDS
A) Optical methods for assaying CaY inhibition properties of compounds:
[00179]
Compounds
of the invention are useful as antagonists of voltage-gated
·ch~neis.
Antagonist properties ~f ~st 'compo~nds ~e;e' ~ssessed as follo~s.
calcium ion
'Cells ~x~ressing
the CaY of interest were placed into microtiter plates_ After an incubation period, the cells were
stained with fluorescent
dyes sensitive to the transmembrane
potential.
The test compounds
were added to the microtiter plate. The cells were stimulated with electrical means to evoke a
Ca V dependent membrane potential change from unblocked channels, which was detected and
measured
with trans-membrane
potential-sensitive
- 232-
dyes.
Antagonists
were detected
as a
PCTfUS2004/036297
WO 2005/042497
~
decreased membrane potential response to the stimulus. The optical membrane potential assay
utilized voltage-sensitive FRET sensors described by Gonzalez and Tsien (See, GonzaIez, J. E.
and R. Y. Tsien (1995) "Voltage sensing by fluorescence resonance energy transfer in single
cells" Biophys J 69(4): 1272-80, and Gonzalez, J. E. and R. Y. Tsien (1997) "Improved
indicators of cell membrane potential that use fluorescence resonance energy transfer" Chern
BioI 4(4): 269-77) in combination with instrumentation for measuring fluorescence changes such
as the Voltagellon Probe Reader (VlPR®) (See, Gonzalez, J. E., K. Oades, et aI. (1999) "Cellbased assays and instrumentation for screening ion-channel targets" Drug Discov Today 4(9):
431-439).
[00180]
VlPR® optical membrane potential assay method with electrical stimulation
[00181]
The following is an example of how CaV2.2 inhibition activity is measured using the
optical membrane potential method. Other subtypes are perfonned in an analogous mode in a
cell line expressing the CaV of interest.
[00182]
HEK293 cells stably expressing CaV2.2 are plated into 96-well microtiter plates.
After an appropriate incubation period, the celIs are stained with the voltage sensitive dyes CC2DMPEJDiSBAC2(3) as follows.
Reagents:
100 mglmL P1uronic F-127 (Sigma #P2443), in dry DMSO
WmM'DiSBAC<;(3) (A~ora 000-100-010) i~ dry DMSO ...•
10·,mM.CC2~DMPE:·(Aurora':,oO-100~8).ili
..
..
-,'
,
"'
'
'
"
clry'DMSO·.·,·. '.' :
','
200 roM Acid Yellow 17 (Aurora #VABSC) in H20
370mM Barium Chloride (Sigma Cat# B6394) in H20
Bath X
160roM NaCl (Sigffia Cat# S-9888)
4.5mM KCI (Sigma Cat# P-5405)
1mM MgCl2 (Fluka Cat# 63064)
10mM HEPES (Sigma Cat# H-4034)
pH 7.4 llsing NaOH
- 233·
.
','
":.'
"
WO 2005/042497
PCTfUS2004/036297
of
Loading rotocol:
[00183]
2X CC2-DMPE = 20 ~M CC2-DMPE:
10 roM CC2-D:MPE is vortexed with· an
equivalent volume of 10% pluronic, followed by vortexing in required amount of HBSS
containing 10 mM HEPES. Each cell plate will require 5 mL of 2X CC2-DMPE, 50 fJ-L of 2X
CC2-DMPE is added to wells containing washed cells, resulting in a 10 fJ-M final staining
concentration. The cells are stained for 30 minutes in the dark at RT.
[00184]
2X CC2DMPE & DISBAC6(3) = 8 p.M CC2DMPE & 2. 5 p.M DISBAC6(3):
Vortex together both dyes with an equivalent volume of 10% pluronic (in DMSO).
Vortex in
required amount of Bath X with beta-cyclodextrin. Each 96well cell plate will require 5 ml of
2XCC2D:MPE. Wash plate with ELx.405 with Bath X, leaving a residual volume of 50 /LUwell.
Add 50 fJ-L of 2XCC2DMPE & DISBAC6(3) to each well. Stain for 30 minutes in the dark at
RT.
[00185]
1. 5X AY17 = 750
J.lM AY17
with 15mM BaCh: Add Acid Yellow 17 to vessel
containing Bath X. Mix well. Allow solution to sit for 10 minutes. Slowly mix in 370mM
BaCh. This solution can be used to solvate compound plates, Note that compound plates are
made at 1.5X drug concentration and not the usual 2X. Wash Ce2 stained plate, again, leaving
residual volume of 50 ILL. Add 100 uUwell of the AY17 solution. Stain forI5 minutes in the
dark at RT. Run plate on the optical reader.
[00186]
The electrical stimulation instrument and methods of use are described in ION
Channel Assay Methods PCrIUS011216.52, herein incoq,oratecf by reference.' .The' instrument .
.
.
.
.':..' '. : .c.o.~pris~s·:a. ~c;~otitet. piate.'.h~d,ler,. 'an':,ppt,icaI
.
'.'
.'
. ,.:'
.,...
.
~
.
.
....
"
~Ystem
for. ex.«iti~g tile .coUlllariri
. '.
.'
.
".
.
:dy~.w.hile. _ .
,.,.'
simultaneously recording the coumarin and oxonol emissions, a waveform generator, a currentor voltage-controlled amplifier, and a device for inserting electrodes in well. Under integrated
computer control, this instrument passes user-programmed electrical stimulus protocols to cells
within the wells of the microtiter plate.
Assay Protoool
[00187]
Insert or use electrodes into each well to be assayed.
[00188]
Use the current-controlled amplifier to deliver stimulation wave pulses for 3-5 s.
Two seconds of pre-stimulus recording are performed to obtain the un-stimulated intensities.
Five seconds of post-stimulation recording are performed to examine the relaxation to the resting
state.
- 234-
WO 20051042497
PCTlUS20041036297
-<
Data Ailalysis
[00189]
Data are analyzed and reported as nonnalized ratios of background-subtracted
emission intensities measured in the 460 nm and 580 nm channels, Background intensities are
then subtracted from each assay channel. Background intensities are obtained by measuring the
emission intensities during the same time periods from identically treated assay wells in which
The response as a function of time is then reported as the ratios obtained
there are no cells.
using the following formula:
(intensity
R(t)
460nm
-
background 460nm)
= -------------------------------(intensity 580 nm - background 580 nm)
[00190]
The data is further reduced by calculating the initial (Ri) and final (Rf) ratios. These
are the average ratio values during part or all of the pre-stimulation period, and during sample
points during the stimulation period. The response to the stimulus
[00191]
R = RflRi is then calculated.
Control responses are obtained by performing assays in the presence of a compound
with the desired properties (positive control), such as mibefradil, and in the absence of
pharmacological agents (negative control).
Responses to the negative (N) and positive (P)
controls are calculated as above. The compound antagonist activity A is defined as:
.' ~='R:-P *100 ..
,. ·N -P ...."
,."
~her~R'is'~er~iiorespons~ofthete~tc~mpOund.
. '".
....'
" ' "
'[00192]'.'EmetRO~ffi:sIOL(j(}y 'AssAyS' Foi'~~,i
A~'
.
.'''p"
.• ,' .","
ANn ~ITIoN'b~':"
TEST COMPOUNDS
[00193]
Patch clamp electrophysiology was used to assess the efficacy of calcium channel
blockers expressed in HEK293 cells. HEK293 cells expressing CaV2.2 have been visually
identified ane!-.probed with fjn~, tip glass el~trodes
connected, to an ~plifier
(Axon.
Instruments). The "voltage clamp" mode has been used to assess the compound's IC50 holding
the cells at - 100 mV. The results of these experiments have contributed to the definition of the
efficacy profile of the compounds.
VOLTAGE-CLAMP assay in HEK293 cells expressing CaV2.2
- 235 -
"',
PCTlUS2004/036297
WO 2005/042497
~
[00194]
CaV2.2 calcium currents were recorded from. HEK293
variation of the patch clamp technique.
with thick walled borosilicate
200B
amplifier
approximately
(Axon
Recordings were made at room temperature (-220 C)
glass electrodes
Instruments).
cells using the whole-cell
(wpI;
After
resistance
establishing
3-4 MQ)
the
using an Axopatch
whole-cell
configuration,
15 minutes were allowed for the pipette solution to equilibrate
before beginning
recording.
Currents
within the cell
2-5 kHz and digitally
were lowpass filtered between
sampled at 10 kHz. Series resistance was compensated 60-70% and was monitored continuously
throughout
the experiment.
The liquid junction
potential
(-7 mY) between the intracellular
pipette solution and the external recording solution was not accounted for in the data analysis.
Test solutions were applied to the cells with a gravity driven fast perfusion system (SF-77;
. Warner Instruments).
[00195]
Dose-response
relationships
were determined
in voltage clamp mode by repeatedly
depolarizing the cell from the experiment specific holding potential to a test potential of +20mV
for 50ms at frequencies of 0.1, 1,5, 10, 15, and 20 Hz. Blocking effects were allowed to plateau
before proceeding to the next test concentration.
[00196]
Solutions
[00197]
Intracellular
(in roM): Cs-F (130), NaCI (10), MgCl2
solution
(1), EGTA (1.5),
CaCl2 (0.1), HEPES (10), glucose (2), pH:: 7.42, 290 mOsm.
[00198]
Extracel~u1ar solu~on (in~:
NaCI (l~8), Ba912 (10), KCI (5.33), KHZP04.(O.44),
.l\1gC;li (D,S), Mg~04·,(Q.41),.·NaHCQ3 (4),. Na2HP04·(O.3), 'gl~C~~e.(5.6):
. . .' .. ~.
.' . ~~.'.
'... .. .
.:
"..
.
:
'
[00199]'
found
'.
':
~es.·(lO)~:. . ...
'."
'Pollowhig these proceatirelf; tepres'eiltatiVe compoundS of the'present
to
possess
desired
N-type
calcium
- 236-
channel
.
invention were·
modulation
activity.
PCTlUS2004/036297
WO 2005/042497
--
[00200]'
Compounds of the invention as shown in Table 2 were found modulate voltage-gated
sodium channels at 25.0 p.M or less.
[00201]
In order that the invention described herein may be more fully understood, the
following examples are set forth. It should be understood that these examples are for illustrative
purposes only and ~e n.otto be construed as limiting this invention in any manner.
-237 -