Note: Descriptions are shown in the official language in which they were submitted.
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METHOD FOR TREATING CHRONIC PAIN
USING MEK INHIBITORS
BACKGROUND
The invention features a method for treating chronic pain. using MEK
inhibitors. Chronic pain includes neuropathic pain, and chronic inflammatory
pain.
Abnormality anywhere in a nerve pathway disrupts nerve signals, which
in turn are abnormally interpreted in the brain, causing neuropathic pain.
Neuropathic pain may be, for example, a deep ache, a burning sensation, or
hypersensitivity to touch. Diseases or conditions associated with neuropathic
pain include, without limitation, diabetic neuropathy, causalgia, plexus
avulsion, neuroma, vasculitis, crush injury, viral infections (e.g., herpes
virus
infection or HIV), constriction injury, tissue injury, nerve injury from the
periphery to the central nervous system, limb amputation, hypothyroidism,
uremia, chronic alcoholism, post-operative pain, arthritis, back pain, and
vitamin deficiencies ,
Infections such as herpes zoster (shingles) can cause nerve
inflammation and produce postherpetic neuralgia, a chronic burning localized
to the area of viral infection. Hyperalgesia is when an already noxious
stimulus becomes more painful, and allodynia, when a previously non-noxious
stimulus becomes painful (such as contact of clothing or a breeze). Reflex
sympathetic dystrophy is accompanied by swelling and sweating or changes
in local blood flow, tissue atrophy, or osteoporosis. Causalgia, including
severe burning pain and swelling, sweating, and changes in blood flow, may
follow an injury or disease of a major nerve such as the sciatic nerve. Some
types of chronic low back pain can have a neuropathic component (e.g.,
sciatica, postpoliomyelitis and CPRM). Neuropathic pain may also be induced
by cancer or chemotherapy.
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Neuropathic pain is currently treated with anticonvulsants such as
carbamazepine and antidepressants such as amitryptaline. NSAIDS and
opioids generally have little effect (Fields et al 1994 Textbook of Pain p 991-
996 (pub: Churchill Livingstone), James & Page 1994
J.Am. Pediatr. Med.Assoc, 8: 439-44 7, Galer, 1995 Neurology 45 S 17-S25.
Neuropathic conditions that have been treated with gabapentin include:
postherpetic neuralgia, postpoliomyelitis, CPRM, HIV-related neuropathy,
trigeminal neuralgia, and reflex sympathetic dystrophy (RSD).
The generally weak efficacy of antiinflammatory agents suggests that the
mechanism for chronic pain is separate from hyperalgesia.
SUMMARY OF THE INVENTION
The invention features a method for treating chronic pain, which
method includes the step of administering a composition including a MEK
inhibitor to a patient in need of such treatment. Chronic pain includes
neuropathic pain, idiopathic pain, and pain associated with vitamin
deficiencies, uremia, hypothyroidism post-operative pain, arthritis, back
pain,
and chronic alcoholism. The invention also features compounds as disclosed,
formulated for the treatment of chronic pain. Such a composition may include
one or more MEK inhibitor compounds having a structure disclosed in patent
applications PCT/US98/13106, international filing date June 24, 1998, and
PCT/US98/13105, international filing date June 24, 1998.
Examples of MEK inhibitors include 4-bromo and 4-iodo.phenylamino
benzhydroxamic acid derivatives which are kinase inhibitors and as such are
useful for treating proliferative diseases such as cancer, psoriasis, and
restenosis. The compounds are defined by Formula I
O R6
~2 C-N-O-R~
R1
N
~ I RS
Br or I
R3 R4
2
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wherein:
R1 is hydrogen, hydroxy, C1-Cg alkyl, C1-Cg alkoxy, halo, trifluoromethyl, or
CN;
R2 is hydrogen;
Rg, R4, and R5 independently are hydrogen, hydroxy, halo, trifluoromethyl,
C1-Cg alkyl, C1-Cg alkoxy, nitro, CN, or (O or NH)m-(CH2)n-Rg, where
Rg is hydrogen, hydroxy, C02H or NR1 OR11
nisOto4;
mis0or1;
R10 and R11 independently are hydrogen or C1-Cg alkyl, or taken together
with the nitrogen to which they are attached can complete a 3- to
10-member cyclic ring optionally containing one, two, or three
additional heteroatoms selected from O, S, NH, or N-C1-Cg alkyl;
O
Rg is hydrogen, C1-Cg alkyl, C-C1-Cg alkyl, aryl, aralkyl, or
C3-C10 cycloalkyl;
R7 is hydrogen, C1-Cg alkyl, C2-Cg alkenyl, C2-Cg alkynyl,
Cg-C10 (cycloalkyl or cycloalkyl optionally containing a heteroatom
selected from O, S, or NRg); or Rg and R~ taken together with the N-O
to which they are attached can complete a 5- to 10-membered cyclic
ring, optionally containing one, two, or three additional heteroatoms
selected from O, S, or NR1 OR11
and wherein any of the foregoing alkyl, alkenyl, and alkynyl groups can be
unsubstituted or substituted by cycloalkyl (or cycloalkyl optionally
containing a
heteroatom selected from O, S, or NRg), aryl, aryloxy, heteroaryl, or
heteroaryloxy.
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Preferred compounds have Formula II
O R6
C-N-O- R~
R1 H
N ~ II
/ I RS
Br or I
R3 R4
where R~, Rg, R4, R5, Rg, and R7 are as defined above. Especially preferred
are compounds wherein R~ is methyl or halo, and Rg, R4, and R5 are halo
such as fluoro or bromo.
Another preferred group of compounds have Formula III
O
ii
1 C - NHOR~
H
N
I I III
/ /
Br or I R3 RS
R4
wherein R~, Rg, R4, R5, and R7 are as defined above.
The most preferred compounds are those wherein R~ is methyl or halo
such as F, Br, CI, and I, R3 is hydrogen or halo such as fluoro, R4 is halo
such as fluoro, and R5 is hydrogen or halo such as fluoro or bromo. Such
compounds have the formulas
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O O
II II
H~ or halo C -NHOR~ CH3 or halo C -NHOR7
H H
\ N \ \ N \
I / I / Br or I / F /
Br or I
F F
O O
II II
CHI or halo C -NHOR~ CH3 or halo C -NHOR7
H H
\ N \ I \ N I \
/ /
Br or 1 I / F I / Br Br or I F F
Specific compounds provided by the invention include the following:
3,4,5-Trifluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;
5-Chloro-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
5-Bromo-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-
benzamide;
N-Hydroxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzamide;
3,4,5-Trifluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;
5-Chloro-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-
benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-
benzamide;
2-(2-Fluoro-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide;
5-Chloro-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-
benzamide;
5-Bromo-2-(2-bromo-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-hydroxy-4-methyl-benzamide;
2-(2-Bromo-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide;
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2-(2-Bromo-4-iodo-phenylamino)-5-chloro-3,4-difluoro-N-hydroxy-
benzamide;
2-(2-Bromo-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide;
4-Fluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;
3,4-Difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;
2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;
2-(2-Bromo-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide;
2-(2-Bromo-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;
N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
5-Bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-
phenylamino)-benzamide;
N-Cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-
benzamide;
N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(2-fluoro-4-iodo-phenylamino)-
benzamide;
5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-
phenylamino)-benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-
3,4-difluoro-benzamide;
N-Cyclopropylmethoxy-2-(2-fluoro-4-iodo-phenylamino)-4-nitro-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-
benzamide;
5-Chloro-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-
3,4-difluoro-benzamide;
5-Bromo-2-(2-bromo-4-iodo-phenylamino)-N-ethoxy-3,4-difluoro-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-ethoxy-4-nitro-benzamide;
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2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-
benzamide;
2-(2-Bromo-4-iodo-phenylamino)-5-chloro-N-cyclopropylmethoxy-
3,4-difluoro-benzamide
2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-nitro-
benzamide;
N-Cyclopropylmethoxy-4-fluoro-2-(2-fluoro-4-iodo-phenylamino)-
benzamide;
N-Cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-
benzamide;
2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-
benzamide;
2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-
benzamide;
4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-N-isopropyl-
benzamide;
N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-N-methyl-
benzamide;
4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-5-nitro-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide;
3,4-Difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;
2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide
(HCI salt);
2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-(tetrahydro-pyran-2-yloxy)-
benzamide;
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3,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)-N-cyclobutylmethoxy-
benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-(2-dimethylamino-ethoxy)-
3,4-difluoro-benzamide monohydrochloride salt;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-
benzamide;
3,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-
benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-
difluoro-benzamide;
5-Bromo-N-cyclohexylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
5-Bromo-N-cyclopentylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide; and
5-Bromo-N-cyclobutylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide.
Other aspects of the invention are provided in the description,
examples and claims below.
Further examples of MEK inhibitors include 4-bromo and 4-iodo
phenylamino benzoic acid derivatives which are selective MEK kinase
inhibitors. The compounds are defined by Formula I(A)
12 Z
R1
N
I RS I (A)
BrorI
R3 R4
wherein:
R~ is hydrogen, hydroxy, C~-Cg alkyl, C~-Cg alkoxy, halo, trifluoromethyl, or
CN;
R2 is hydrogen;
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Rg, R4, and R5 independently are hydrogen, hydroxy, halo, trifluoromethyl,
C1-Cg alkyl, C1-Cg alkoxy, nitro, CN, or -(O or NH)m -(CH2)n-Rg,
where Rg is hydrogen, hydroxy, C02H, or NR1pR11;
n is 0-4;
mis0or1;
R1 p and R11 independently are hydrogen or C1-Cg alkyl, or taken together
with the nitrogen to which they are attached, can complete a
3-10 member cyclic ring optionally containing one, two, or three
additional heteroatoms selected from O, S, NH, or N-C1-Cg alkyl;
Z is COOR7, tetrazolyl, CONRgR7, CONHNR1pR11, or CH20R7;
Rg and R~ independently are hydrogen, C1-Cg alkyl, C2-Cg alkenyl,
O
C2-Cg alkynyl, C - C1-Cg alkyl, aryl, heteroaryl,
C3-C10 cycloalkyl, or C3-C10 (cycloalkyl optionally containing
one, two, or three heteroatoms selected from O, S, NH, or N
alkyl); or Rg and R~ together with the nitrogen to which they are
attached complete a 3-10 member cyclic ring optionally
containing 1, 2, or 3 additional heteroatoms selected from O, S,
NH, or N alkyl;
and wherein any of the foregoing alkyl, alkenyl, and alkynyl groups can be
unsubstituted or substituted by halo, hydroxy, alkoxy, amino,
alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, heteroaryl, or
heteroaryloxy, and the pharmaceutically acceptable salts thereof.
Preferred compounds have Formula II(A)
O
C-O-R~
R1 H
I I (A)
I Rs
Br or I v \
R3 R4
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where R1, R3, R4, R5, Rg, and R7 are as defined above. Especially preferred
are compounds wherein R1 is methyl or halo, and R3, R4, and R5 are halo
such as fluoro or bromo.
The compounds of Formula II(A) are carboxylic acids when R7 is
hydrogen, and are esters when R7 is other than hydrogen. Compounds which
are analogous to the acids in physical and biological properties are
tetrazolyl
derivatives of Formula Ila
N-N
i
N ~ NCR
9
R1 H
N
/ I RS or
Br or I
R3 R4
~ R9
N-N
ii
N ,N
R1 H
N
I RS
Br or I
R3 R4
Another preferred group of compounds are amides Formula III(A)
Ila
O
~i
R C - NR6R~
1 H
N
I I III(A)
/ /
Br or I R3 RS
R4
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and hydrazides of Formula Illa
O
ii
R C -NHNR1~R11
1 H
N
Illa
I / I
Br or I ~' R3 ~ \R5
R4
The benzyl alcohols of the invention have Formula IV(A)
H20R~
R1 H
N
I RS IV(A)
Br or I
R3 R4
Among this group, the most preferred compounds are those wherein
R1 is methyl, R3 is hydrogen or halo such as fluoro, R4 is halo such as
fluoro,
and R5 is hydrogen or halo such as fluoro, bromo, or chloro. Representative
compounds have the formulas
CH3 Z CH3 Z
H H
\ N \ \ N \
/ / / /
Br or I Br or I F
F F
CH3 Z CH3 Z
H H
\ N \ ~ \ N \
/ ~ / Cl gr or I / F / F
Br or I F ~Br
F F F
11
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Preferred embodiments for this invention include methods using one or
more of the following compounds:
(a) said MEK inhibitor has a structure selected from:
2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-
benzamide;
N-Cyclopropylmethoxy-3,4,5-trifuoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
N-Cyclopropylmethoxy-3,4,5-trifuoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide, potassium salt;
2-(2-Chloro-4-iodo-phenylamino)-N-cyclobutylmethoxy-3,4-difluoro-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-
benzamide;
5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-methoxy-
benzamide;
3,4-Difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-
benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-
benzamide;
N-Cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
5-Bromo-N-cyclobutylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
5-Bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
5-Chloro-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-
difluoro-benzamide;
4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;
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4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide,
hydrochloride salt;
5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-(2-hydroxy-ethoxy)-
benzamide;
3,4-Difluoro-N-(2-hydroxy-ethoxy)-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-(3-hydroxy-
propoxy)-benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4,5-trifluoro-N-(3-hydroxy-propoxy)-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4,5-trifluoro-N-[2-(2-methoxy-
ethoxy)-ethoxy]-benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-(3-hydroxy-propoxy)-
benzamide;
5-Bromo-3,4-difluoro-N-(3-hydroxy-propoxy)-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
3,4,5-Trifluoro-N-(3-hydroxy-propoxy)-2-(4-iodo-2-methyl-
phenylamino)-benzamide;
3,4,5-Trifluoro-N-(2-hydroxy-ethoxy)-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-(2-hydroxy-ethoxy)-
benzamide; and
3,4-Difluoro-N-(2-hydroxy-ethoxy)-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
(b) said MEK inhibitor has a structure selected from:
2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-
benzamide;
N-Cyclopropylmethoxy-3,4,5-trifuoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-(2-hydroxy-ethoxy)-
benzamide;
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3,4-Difluoro-N-(2-hydroxy-ethoxy)-2-(4-iodo-2-methyl-phenylamino)-
benzamide;
(c) said MEK inhibitor has a structure selected from:
2-(2-Chloro-4-iodo-phenylamino)-3,4difluoro-benzoic acid;
3,4,5-Trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid;
5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzoic acid;
3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid;
5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid;
2-(2-Chloro-4-iodo-pyenylamino)-3,4-difluoro-5-nitro-benzoic acid;
2-(2-Chloro-4-iodo-phenylamino)-3,4,5-trifluoro-benzoic acid;
7-Fluoro-6-(4-iodo-2-methyl-phenylamino)1 H-benzoimidazole-5-
carboxylic acid cyclopropylmethoxy-amide;
5-Chloro-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid;
and
5-Chloro-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzoic acid;
and
(d) said MEK inhibitor has a structure selected from:
2-(2-Chloro-4-iodo-phenylamino)-3,4difluoro-benzoic acid; and
7-Fluoro-6-(4-iodo-2-methyl-phenylamino)1 H-benzoimidazole-5-
carboxylic acid cyclopropylmethoxy-amide.
This invention also provides pharmaceutical formulations adapted for
the treatment of chronic pain, said formalities comprising a disclosed
compound together with a pharmaceutically acceptable excipient, diluent, or
carrier. Preferred formulations include any of the foregoing preferred
compounds together with an excipient, diluent, or carrier.
The disclosed compounds are potent and selective inhibitors of kinase
enzymes, particularly MEK1 and MEK2.
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BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 is a bar graph representing the paw withdrawal threshold (PWT)
in grams as a function of time in days. The empty, cross-hatched, and single-
s hatched bars are vehicle, PD 198306, and pregabalin, respectively. The
arrows indicate time of drug administration (30 mg/kg, p.o.).
FIG 2. is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments as a function of time in days.
Baseline (BL) measurements were taken before treatment. Animals received
a single p.o. administration of PD 198306 (3-30mg/kg), or pregabalin
(30mg/kg) and withdrawal thresholds were re-assessed 1 h after treatment.
Treatments were repeated twice a day for two days. Results are expressed
median ~ 1St and 3~d quartiles. *P<0.05, **P<0.01, ***P<0.001 significantly
different from vehicle treated animals (Mann-Whitney t test; n=7-8).
FIG. 3. is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments as a function of time in days.
Baseline (BL) measurements were taken before treatment. Animals received
a single p.o. administration of PD 198306 (3-30mg/kg), or pregabalin
(30mg/kg) and withdrawal thresholds were re-assessed 1 h after treatment.
Treatments were repeated twice a day for two days. Results are expressed
median ~ 1St and 3~d quartiles. **P<0.01 significantly different from vehicle
treated animals (Mann-Whitney t test; n=6).
FIG. 4. is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments as a function of time in days.
Baseline (BL) measurements were taken before treatment. Animals received
a single i.t. administration of PD 198306 (1-30wg/10w1), or pregabalin
(100p,g/10~1) and withdrawal thresholds were re-assessed at 30min, 1 h and
2h after treatment. Results are expressed median ~ 1 St and 3'd quartiles.
*P<0.05, ***P<0.001 significantly different from vehicle treated animals (Mann-
Whitney t test; n=7-9).
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FIG. 5. is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments as a function of time in days.
Baseline (BL) measurements were taken before treatment. Animals received
a single i.t. administration of PD 198306 (1-30pg/10p1), or pregabalin
(100~g/10p.1) and withdrawal thresholds were re-assessed at 30min, 1 h and
2h after treatment. Results are expressed median ~ 1St and 3'd quartiles.
*P<0.05, **P<0.01, ***P<0.001 significantly different from vehicle treated
animals (Mann-Whitney t test; n=6-8).
FIG. 6 is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments as a function of time in days .
Animals received a single intraplantar (i.pl.) administration of PD 198306
(3mg/100p,1), or an intrathecal injection of PD 198306 (30pg/10~,1) and
withdrawal thresholds were re-assessed 1 h after treatment. Results are
expressed median ~ 1St and 3'd quartiles. **P<0.01 significantly different
from
vehicle treated animals (Mann-Whitney t test; n=6-9).
FIG. 7. is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments as a function of time in days.
Animals received a single intraplantar (i.pl.) administration of PD 198306
(3mg/100~1), or an intrathecal injection of PD 198306 (30~.g/10p1) and
withdrawal thresholds were re-assessed 1 h after treatment. Results are
expressed median ~ 1St and 3'd quartiles. **P<0.01 significantly different
from
vehicle treated animals (Mann-Whitney t test; n=6).
FIG. 8 is a bar graph representing the force required in grams to elicit
paw withdrawal using von Frey hair filaments. Baseline (BL) measurements
were taken before treatment. Animals received a single i.t. administration of
PD219622, PD297447, PD 184352, or PD 254552 (30pg/10~1), or pregabalin
(100~g/10p1) and withdrawal thresholds were re-assessed at 30min, 1 h and
2h after treatment. Results are expressed median ~ 1St and 3'd quartiles.
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*P<0.05, **P<0.01, ***P<0.001 significantly different from vehicle treated
animals (Mann-Whitney t test; n=7-8).
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DETAILED DESCRIPTION
The compounds disclosed herein are pharmaceutically active, for
example, they inhibit MEK. MEK enzymes are dual specificity kinases
involved in, for example, immunomodulation, inflammation, and proliferative
diseases such as cancer and restenosis, as well as pair.
Proliferative diseases are caused by a defect in the intracellular
signaling system, or the signal transduction mechanism of certain proteins.
Defects include a change either in the intrinsic activity or in the cellular
concentration of one or more signaling proteins in the signaling cascade . The
cell may produce a growth factor that binds to its own receptors, resulting in
an autocrine loop, which continually stimulates proliferation. Mutations or
overexpression of intracellular signaling proteins can lead to spurious
mitogenic signals within the cell. Some of the most common mutations occur
in genes encoding the protein known as Ras, a G-protein that is activated
when bound to GTP, and inactivated when bound to GDP. The above-
mentioned growth factor receptors, and many other mitogenic receptors, when
activated, lead to Ras being converted from the GDP-bound state to the GTP-
bound state. This signal is an absolute prerequisite for proliferation in most
cell types. Defects in this signaling system, especially in the deactivation
of
the Ras-GTP complex, are common in cancers, and lead to the signaling
cascade below Ras being chronically activated.
Activated Ras leads in turn to the activation of a cascade of
serine/threonine kinases. One of the groups of kinases known to require an
active Ras-GTP for its own activation is the Raf family. These in turn
activate
MEK (e.g., MEK1 and MEK2) which then activates MAP kinase, ERK (ERK~
and ERK2). Activation of MAP kinase by mitogens appears to be essential for
proliferation; constitutive activation of this kinase is sufficient to induce
cellular
transformation. Blockade of downstream Ras signaling, for example by use of
a dominant negative Raf-1 protein, can completely inhibit mitogenesis,
whether induced from cell surface receptors or from oncogenic Ras mutants.
Although Ras is not itself a protein kinase, it participates in the activation
of
Raf and other kinases, most likely through a phosphorylation mechanism.
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Once activated, Raf and other kinases phosphorylate MEK on two closely
adjacent serine residues, S218 and S222 in the case of MEK-1, which are the
prerequisite for activation of MEK as a kinase. MEK in turn phosphorylates
MAP kinase on both a tyrosine, Y185, and a threonine residue, T183,
separated by a single amino acid.
This double phosphorylation activates MAP kinase at least 100-fold.
Activated MAP kinase can then catalyze the phosphorylation of a large
number of proteins, including several transcription factors and other kinases.
Many of these MAP kinase phosphorylations are mitogenically activating for
the target protein, such as a kinase, a transcription factor, or another
cellular
protein. In addition to Raf-1 and MEKK, other kinases activate MEK, and
MEK itself appears to be a signal integrating kinase. Current understanding
is that MEK is highly specific for the phosphorylation of MAP kinase. In fact,
no substrate for MEK other than the MAP kinase , ERK, has been
demonstrated to date and MEK does not phosphorylate peptides based on
the MAP kinase phosphorylation sequence, or even phosphorylate denatured
MAP kinase. MEK also appears to associate strongly with MAP kinase prior
to phosphorylating it, suggesting that phosphorylation of MAP kinase by MEK
may require a prior strong interaction between the two proteins. Both this
requirement and the unusual specificity of MEK are suggestive that it
may have enough difference in its mechanism of action to other protein
kinases that selective inhibitors of MEK, possibly operating through
allosteric
mechanisms rather than through the usual blockade of the ATP binding site,
may be found.
The effect of the MEK inhibitor PD 198306 has been investigated in two
animal models of neuropathic pain by assessing static allodynia with von Frey
hairs.
Oral administration of PD 198306 (3-30mg/kg) had no effect in the model
of chronic constriction injury of the sciatic nerve (CCI). However, after
repeated
administration (3 doses over two days) it had a transient effect in the
diabetic
neuropathy model (streptozocin). This may be due to disorders of the blood-
brain barrier induced by the diabetic condition in these animals, thus
allowing
central action of the compound. Intrathecal administration of PD 198306 (1-
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30~g) dose-dependently blocked static allodynia in both the streptozocin and
the
CCI models of neuropathic pain, with minimum effective doses (MED) of 3 and
10~g respectively. The highest dose used (30~g) totally blocked the
maintenance of static allodynia, for up to 1 h. Intraplantar administration of
PD
198306 (3mg/100~1) at a dose 100-fold higher than the dose shown to be
effective intrathecally (30~g/10~1) had no effect on static allodynia in
either of the
neuropathic pain models. This finding confirms the. lack of effect seen after
systemic administration and suggests a central site of action for the
compound.
This study supports the use of MEK inhibitors as potential new
therapeutic tools for chronic pain. The study of potential side-effects,
especially related to memory, of future brain-penetrant MEK inhibitors will
further support the therapeutic window for this novel class of compounds in
the treatment of pain.
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A. Terms
Certain terms are defined below and by their usage throughout this
disclosure.
Alkyl groups include aliphatic (i.e., hydrocarbyl or hydrocarbon radical
structures containing hydrogen and carbon atoms) with a free valence. Alkyl
groups are understood to include straight chain and branched structures.
Examples include methyl, ethyl, propyl, isopropyl, butyl, n-butyl, isobutyl, t-
butyl, pentyl, isopentyl, 2,3-dimethylpropyl, hexyl, 2,3-dimethylhexyl, 1,1-
dimethylpentyl, heptyl, and octyl. Cycloalkyl groups include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.
Alkyl groups can be substituted with 1, 2, 3 or more substituents which
are independently selected from halo (fluoro, chloro, bromo, or iodo),
hydroxy,
amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy,
arylalkyloxy,
heterocyclic radical, and (heterocyclic radical)oxy. Specific examples include
fluoromethyl, hydroxyethyl, 2,3-dihydroxyethyl, (2- or 3-furanyl)methyl,
cyclopropylmethyl, benzyloxyethyl, (3-pyridinyl)methyl, (2- or 3-
furanyl)methyl,
(2-thienyl)ethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl,
methoxymethyl, N-pyridinylethyl, diethylaminoethyl, and cyclobutylmethyl.
Alkenyl groups are analogous to alkyl groups, but have at least one
double bond (two adjacent sp2 carbon atoms). Depending on the placement
of a double bond and substituents, if any, the geometry of the double bond
may be entgegen (E), or zusammen (Z), cis, or trans. Similarly, alkynyl
groups have at least one triple bond (two adjacent sp carbon atoms).
Unsaturated alkenyl or alkynyl groups may have one or more double or triple
bonds, respectively, or a mixture thereof; like alkyl groups, unsaturated
groups
may be straight chain or branched, and they may be substituted as described
both above for alkyl groups and throughout the disclosure by example.
Examples of alkenyls, alkynyls, and substituted forms include cis-2-butenyl,
trans-2-butenyl, 3-butynyl, 3-phenyl-2-propynyl, 3-(2'-fluorophenyl)-2-
propynyl,
3-methyl(5-phenyl)-4-pentynyl, 2-hydroxy-2-propynyl, 2-methyl-2-propynyl, 2-
propenyl, 4-hydroxy-3-butynyl, 3-(3-fluorophenyl)-2-propynyl, and 2-methyl-2-
propenyl. In formula (I), alkenyls and alkynyls can be C 2~ or C 2_$, for
example, and are preferably C 3_4 or C 3_8.
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More general forms of substituted hydrocarbon radicals include
hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl,
and corresponding forms for the prefixes amino-, halo- (e.g., fluoro-, chloro-
,
or bromo-), nitro-, alkyl-, phenyl-, cycloalkyl- and so on, or combinations of
substituents. According to formula (I), therefore, substituted alkyls include
hydroxyalkyl, aminoalkyl, nitroalkyl, haloalkyl, alkylalkyl (branched alkyls,
such
as methylpentyl), (cycloalkyl)alkyl, phenylalkyl, alkoxy, alkylaminoalkyl,
dialkylaminoalkyl, arylalkyl, aryloxyalkyl, arylalkyloxyalkyl, (heterocyclic
radical)alkyl, and (heterocyclic radical)oxyalkyl. R~ thus includes
hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl, hydroxyaryl,
aminoalkyl, aminoalkenyl, aminoalkynyl, aminocycloalkyl, aminoaryl,
alkylalkenyl, (alkylaryl)alkyl, (haloaryl)alkyl, (hydroxyaryl)alkynyl, and so
forth.
Similarly, RA includes hydroxyalkyl and aminoaryl, and RB includes
hydroxyalkyl, aminoalkyl, and hydroxyalkyl(heterocyclic radical)alkyl.
Heterocyclic radicals, which include but are not limited to heteroaryls,
include: furyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, pyrrolyl,
imidazolyl,
1,3,4-triazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, indolyl, and
their
nonaromatic counterparts. Further examples of heterocyclic radicals include
piperidyl, quinolyl, isothiazolyl, piperidinyl, morpholinyl, piperazinyl,
tetrahydrofuryl, tetrahydropyrrolyl, pyrrolidinyl, octahydroindolyl,
octahydrobenzothiofuranyl, and octahydrobenzofuranyl.
Selective MEK 1 or MEK 2 inhibitors are those compounds which
inhibit the MEK 1 or MEK 2 enzymes, respectively, without substantially
inhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase,
EGF, and PDGF receptor kinases, and C-src. In general, a selective MEK 1
or MEK 2 inhibitor has an ICSO for MEK 1 or MEK 2 that is at least one-
fiftieth
(1/50) that of its ICSO for one of the above-named other enzymes. Preferably,
a selective inhibitor has an ICSO that is at least 1/100, more preferably
1/500,
and even more preferably 1/1000, 1/5000, or less than that of its ICSO or one
or
more of the above-named enzymes.
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B. Compounds
As used herein, the term "aryl" means a cyclic, bicyclic, or tricyclic
aromatic ring moiety having from five to twelve carbon atoms. Examples of
typical aryl groups include phenyl, naphthyl, and fluorenyl. The aryl may be
substituted by one, two, or three groups selected from fluoro, chloro, bromo,
iodo, alkyl, hydroxy, alkoxy, nitro, or amino. Typical substituted aryl groups
include 3-fluorophenyl, 3,5-dimethoxyphenyl, 4-nitronaphthyl, 2-methyl-
4-chloro-7-aminofluorenyl, and the like.
The term "aryloxy" means an aryl group bonded through an oxygen
atom, for example phenoxy, 3-bromophenoxy, naphthyloxy, and 4-methyl-
1-fluorenyloxy.
"Heteroaryl" means a cyclic, bicyclic, or tricyclic aromatic ring moiety
having from four to eleven carbon atoms and one, two, or three heteroatoms
selected from O, S, or N. Examples include furyl, thienyl, pyrrolyl,
pyrazolyl,
triazolyl, thiazolyl, xanthenyl, pyronyl, indolyl, pyrimidyl, naphthyridyl,
pyridyl,
and triazinyl. The heteroaryl groups can be unsubstituted or substituted by
one, two, or three groups selected from fluoro, chloro, bromo, iodo, alkyl,
hydroxy, alkoxy, nitro, or amino. Examples of substituted heteroaryl groups
include chloropyranyl, methylthienyl, fluoropyridyl, amino-1,4-benzisoxazinyl,
nitroisoquinolinyl, and hydroxyindolyl.
The heteroaryl groups can be bonded through oxygen to make
heteroaryloxy groups, for example thienyloxy, isothiazolyloxy,
benzofuranyloxy, pyridyloxy, and 4-methylisoquinolinyloxy.
The term "C1-Cg alkyl" means straight and branched chain aliphatic
groups having from one to eight carbon atoms. Typical C1-Cg alkyl groups
include methyl, ethyl, isopropyl, tert.-butyl, 2,3-dimethylhexyl, and
1,1-dimethylpentyl. The alkyl groups can be unsubstituted or substituted by
cycloalkyl, cycloalkyl containing a heteroatom selected from O, S, or NRg,
aryl, aryloxy, heteroaryl, or heteroaryloxy, as those terms are defined above.
Examples of aryl and aryloxy substituted alkyl groups include phenylmethyl,
2-phenylethyl, 3-chlorophenylmethyl, 1,1-dimethyl-3-(2-nitrophenoxy)butyl,
and 3,4,5-trifluoronaphthylmethyl. Examples of alkyl groups substituted by a
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heteroaryl or heteroaryloxy group include thienylmethyl, 2-furylethyl,
6-furyloxyoctyl, 4-methylquinolyloxymethyl, and 6-isothiazolylhexyl.
Cycloalkyl
substituted alkyl groups include cyclopropylmethyl, 2-cyclopentylethyl,
2-piperidin-1-ylethyl, 3-(tetrahydropyran-2-yl)propyl, and cyclobutylmethyl.
"C2-Cg Alkenyl" means a straight or branched carbon chain having one
or more double bonds. Examples include but-2-enyl, 2-methyl-prop-2-enyl,
1,1-dimethyl-hex-4-enyl, 3-ethyl-4-methyl-pent-2-enyl, and 3-isopropyl-pent-
4-enyl. The alkenyl groups can be substituted with aryl, aryloxy, heteroaryl,
or
heteroyloxy, for example 3-phenylprop-2-enyl, 6-thienyl-hex-2-enyl, 2-furyloxy-
but-2-enyl, and 4-naphthyloxy-hex-2-enyl.
"C2-Cg Alkynyl" means a straight or branched carbon chain having
from two to eight carbon atoms and at least one triple bond. Typical alkynyl
groups include prop-2-ynyl, 2-methyl-hex-5-ynyl, 3,4-dimethyl-hex-5-ynyl, and
2-ethyl-but-3-ynyl. The alkynyl groups can be substituted by aryl, aryloxy,
heteroaryl, or heteroaryloxy, for example 4-(2-fluorophenyl)-but-3-ynyl,
3-methyl-5-thienylpent-4-ynyl, 3-phenoxy-hex-4-ynyl, and 2-furyloxy-3-methyl-
hex-4-ynyl.
The alkenyl and alkynyl groups can have one or more double bonds or
triple bonds, respectively, or a combination of double and triple bonds. For
example, typical groups having both double and triple bonds include hex-2-en-
4-ynyl, 3-methyl-5-phenylpent-2-en-4-ynyl, and 3-thienyloxy-hex-3-en-5-ynyl.
The term "Cg-C1 p cycloalkyl" means a non-aromatic ring or fused rings
containing from three to ten carbon atoms. Examples include cyclopropyl,
cyclobutyl, cyclopenyl, cyclooctyl, bicycloheptyl, adamantyl, and cyclohexyl.
The ring can optionally contain a heteroatom selected from O, S, or NRg.
Such groups include tetrahydrofuryl, tetrahydropyrrolyl,
octahydrobenzofuranyl, octahydroindolyl, and octahydrobenzothiofuranyl.
R3, R4, and R5 can include groups defined by the term (O or NH)m-
(CH2)n-Rg. Examples of such groups are aminomethyl, 2-aminoethyl,
2-aminoethylamino, 3-aminopropoxy, N,N-diethylamino, 3-(N-methyl-N-
isopropylamino)-propylamino, 2-(N-acetylamino)-ethoxy, 4-(N-
dimethylaminocarbonylamino)-butoxy, and 3-(N-cyclopropylamino)-propoxy.
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As used herein, the term "aryl" means a cyclic, bicyclic, or tricyclic
aromatic ring moiety having from five to twelve carbon atoms. Examples of
typical aryl groups include phenyl, naphthyl, and fluorenyl. The aryl may be
substituted by one, two, or three groups selected from fluoro, chloro, bromo,
iodo, alkyl, hydroxy, alkoxy, nitro, amino, alkylamino, or dialkylamino.
Typical
substituted aryl groups include 3-fluorophenyl, 3,5-dimethoxyphenyl,
4-nitronaphthyl, 2-methyl-4-chloro-7-aminofluorenyl, and the like.
The term "aryloxy" means an aryl group bonded through an oxygen
atom, for example phenoxy, 3-bromophenoxy, naphthyloxy, and 4-methyl-
1-fluorenyloxy.
"Heteroaryl" means a cyclic, bicyclic, or tricyclic aromatic ring moiety
having from four to eleven carbon atoms and one, two, or three heteroatoms
selected from O, S, or N. Examples include furyl, thienyl, pyrrolyl,
pyrazolyl,
imidazolyl, triazolyl, thiazolyl, oxazolyl, xanthenyl, pyronyl, indolyl,
pyrimidyl,
naphthyridyl, pyridyl, benzinnidazolyl, and triazinyl. The heteroaryl groups
can -
be unsubstituted or substituted by one, two, or three groups selected from
fluoro, chloro, bromo, iodo, alkyl, hydroxy, alkoxy, nitro, amino, alkylamino,
or
dialkylamino. Examples of substituted heteroaryl groups include
chloropyranyl, methylthienyl, fluoropyridyl, amino-1,4-benzisoxazinyl,
nitroisoquinolinyl, and hydroxyindolyl.
The heteroaryl groups can be bonded through oxygen to make
heteroaryloxy groups, for example thienyloxy, isothiazolyloxy,
benzofuranyloxy, pyridyloxy, and 4-methylisoquinolinyloxy.
The term "C1-Cg alkyl" means straight and branched chain aliphatic
groups having from one to eight carbon atoms, preferably one to four. Typical
C1-Cg alkyl groups include methyl, ethyl, isopropyl, tert.-butyl,
2,3-dimethylhexyl, and 1,1-dimethylpentyl. The alkyl groups can be
unsubstituted or substituted by halo, hydroxy, alkoxy, amino, alkylamino,
dialkylamino, cycloalkyl, aryl, aryloxy, heteroaryl, or heteroaryloxy, as
those
terms are defined herein. Typical substituted alkyl groups include
chloromethyl, 3-hydroxypropyl, 2-dimethylaminobutyl, and
2-(hydroxymethylamino)ethyl. Examples of aryl and aryloxy substituted alkyl
groups include phenylmethyl, 2-phenylethyl, 3-chlorophenylmethyl,
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1,1-dimethyl-3-(2-nitrophenoxy)butyl, and 3,4,5-trifluoronaphthylmethyl.
Examples of alkyl groups substituted by a heteroaryl or heteroaryloxy group
include thienylmethyl, 2-furylethyl, 6-furyloxyoctyl, 4-
methylquinolyloxymethyl,
and 6-isothiazolylhexyl. Cycloalkyl substituted alkyl groups include
cyclopropylmethyl, 2-cyclohexyethyl, piperidyl-2-methyl, 2-(piperidin-1-yl)-
ethyl, 3-(morpholin-4-yl)propyl.
"C2-Cg Alkenyl" means a straight or branched carbon chain having one
or more double bonds. Examples include but-2-enyl, 2-methyl-prop-2-enyl,
1,1-dimethyl-hex-4-enyl, 3-ethyl-4-methyl-pent-2-enyl, and 3-isopropyl-pent-
4-enyl. The alkenyl groups can be substituted with halo, hydroxy, alkoxy,
amino, alkylamino, dialkylamino, aryl, aryloxy, heteroaryl, or heteroyloxy,
for
example 2-bromoethenyl, 3-hydroxy-2-butenyl, 1-aminoethenyl, 3-phenylprop-
2-enyl, 6-thienyl-hex-2-enyl, 2-furyloxy-but-2-enyl, and 4-naphthyloxy-hex-
2-enyl.
"C2-Cg Alkynyl" means a straight or branched carbon chain having
from two to eight carbon atoms and at least one triple bond. Typical alkynyl
groups include prop-2-ynyl, 2-methyl-hex-5-ynyl, 3,4-dimethyl-hex-5-ynyl, and
2-ethyl-but-3-ynyl. The alkynyl groups can be substituted as the alkyl and
alkenyl groups, for example, by aryl, aryloxy, heteroaryl, or heteroaryloxy,
for
example 4-(2-fluorophenyl)-but-3-ynyl, 3-methyl-5-thienylpent-4-ynyl,
3-phenoxy-hex-4-ynyl, and 2-furyloxy-3-methyl-hex-4-ynyl.
The alkenyl and alkynyl groups can have one or more double bonds or
triple bonds, respectively, or a combination of double and triple bonds. For
example, typical groups having both double and triple bonds include hex-2-en-
4-ynyl, 3-methyl-5-phenylpent-2-en-4-ynyl, and 3-thienyloxy-hex-3-en-5-ynyl.
The term "C3-C1 p cycloalkyl" means a nonaromatic ring or fused rings
containing from three to ten carbon atoms. Examples include cyclopropyl,
cyclobutyl, cyclopenyl, cyclooctyl, bicycloheptyl, adamantyl, and cyclohexyl.
The ring can optionally contain one, two, or three heteroatoms selected from
O, S, or NRg. Such groups include tetrahydrofuryl, tetrahydropyrrolyl,
octahydrobenzofuranyl, morpholinyl, piperazinyl, pyrrolidinyl, piperidinyl,
octahydroindolyl, and octahydrobenzothiofuranyl. The cycloalkyl groups can
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be substituted with the same substituents as an alkyl and alkenyl groups, for
example, halo, hydroxy, aryl, and heteroaryloxy. Examples include
3-hydroxycyclohexyl, 2-aminocyclopropyl, 2-phenylpyrrolidinyl, and
3-thienylmorpholine-1-yl.
S Rg and R7 can be taken together with the nitrogen to which they are
attached to complete a cyclic ring having from 3 to 10 members, which
may contain 1, 2, or 3 additional heteroatoms selected from O, S, NH, or N
alkyl. Examples of such cyclic rings include piperazinyl, piperidyl,
pyrrolidinyl,
morpholino, N-methylpiperazinyl, aziridynyl, and the like. Such rings can be
substituted with halo, hydroxy, alkyl, alkoxy, amino, alkyl, and dialkylamino,
aryl, aryloxy, heteroaryl, and heteroaryloxy. Typical examples include
3-hydroxy-pyrrolidinyl, 2-fluoro-piperindyl, 4-(2-hydroxyethyl)-piperidinyl,
and
3-thienylmorpholino.
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C. Synthesis
The 4-bromo and 4-iodo phenylamino benzhydroxamic acid derivatives
of Formula I can be prepared from commercially available starting materials
utilizing synthetic methodologies well-known to those skilled in organic
chemistry. A typical synthesis is carried out by reacting a 4-bromo or 4-iodo
aniline with a benzoic acid having a leaving group at the 2-position to give a
phenylamino benzoic acid, and then reacting the benzoic acid phenylamino
derivative with a hydroxylamine derivative. This process is depicted in
Scheme 1.
I0 Scheme 1
O
II
~2 C-OH
R1
\ NH L \
RS
Br or I
R3 R4
base
O
R II
R1 I2 C-OH
\ N \
/ I RS
Br or I
R3 R4
R6
HN-O-R~
R O R6
C-N-O-R
Rl I 7
\ N \
/ I RS
Br or I
R3 R4
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where L is a leaving group, for example halo such as fluoro, chloro, bromo or
iodo, or an activated hydroxy group such as a diethylphosphate,
trimethylsilyloxy, p-nitrophenoxy, or phenylsulfonoxy.
The reaction of the aniline derivative and the benzoic acid derivative
generally is accomplished by mixing the benzoic acid with an equimolar
quantity or excess of the aniline in an unreactive organic solvent such as
tetrahydrofuran, or toluene, in the presence of a base such as lithium
diisopropylamide, n-butyl lithium, sodium hydride, and sodium amide. The
reaction generally is carried out at a temperature of about -78°C to
about
25°C, and normally is complete within about 2 hours to about 4 days.
The
product can be isolated by removing the solvent, for example by evaporation
under reduced pressure, and further purified, if desired, by standard methods
such as chromatography, crystallization, or distillation.
The phenylamino benzoic acid next is reacted with a hydroxylamine
derivative HNRgOR7 in the presence of a peptide coupling reagent.
Hydroxylamine derivatives that can be employed include methoxylamine,
N-ethyl-isopropoxy amine, and tetrahydro-oxazine. Typical coupling reagents
include 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ),
1,3-dicyclohexylcarbodiimide (DCC), bromo-tris(pyrrolidino)-phosphonium
hexafluorophosphate (PyBrOP) and (benzotriazolyloxy)tripyrrolidino
phosphonium hexafluorophosphate (PyBOP). The phenylamino benzoic acid
and hydroxylamino derivative normally are mixed in approximately equimolar
quantities in an unreactive organic solvent such as dichloromethane,
tetrahydrofuran, chloroform, or xylene, and an equimolar quantity of the
coupling reagent is added. A base such as triethylamine or
diisopropylethylamine can be added to act as an acid scavenger if desired.
The coupling reaction generally is complete after about 10 minutes to 2 hours,
and the product is readily isolated by removing the reaction solvent, for
instance by evaporation under reduced pressure, and purifying the product by
standard methods such as chromatography or crystallizations from solvents
such as acetone, diethyl ether, or ethanol.
An alternative method for making the invention compounds involves
first converting a benzoic acid to a hydroxamic acid derivative, and then
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reacting the hydroxamic acid derivative with an aniline. This synthetic
sequence is depicted in Scheme 2.
Scheme 2
O O R6
C-OH R C-N-O-R~
16
L \ HN_O-R7 L \
RS I RS
v
R3 R4 R3 R4
NHR2
I R1
Br or I
R O R6
R I2 C-N-O-R~
1
\ N \
/ I RS
Br or I
R3 R4
where L is a leaving group. The general reaction conditions for both of the
steps in Scheme 2 are the same as those described above for Scheme 1.
Yet another method for making invention compounds comprises
reacting a phenylamino benzhydroxamic acid with an ester forming group as
depicted in Scheme 3.
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Scheme 3
R O R6
R I2 C-N-OH
1
N
/ I RS + L - R~
BrorI v
R3 R4
base
O R6
C-N-O-R~
R1
N
I RS
Br or I
R3 R4
where L is a leaving group such as halo, and a base is triethylamine or
diisopropylamine.
The synthesis of invention compounds of Formula I is further illustrated
by the following detailed examples numbers 1 to 102.
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The 2-(4-bromo and 4-iodo phenylamino)-benzoic acid derivatives of
Formula I(A) can be prepared from commercially available starting materials
utilizing synthetic methodologies well-known to those skilled in organic
chemistry and illustrated in synthetic examples 1A - 224A below. A typical
synthesis is carried out by reacting a 4-bromo or 4-iodo aniline with a
benzoic
acid having a leaving group at the 2-position to give a 2-(phenylamino)-
benzoic acid. This process is depicted in Scheme 1 (A).
Scheme 1 (A)
O
R II
R1 I 2 C-OH
\ NH L \
I RS
BrorI
R3 R4
base
O
I I
R i 2 C-OH
1
\ N \
/ I Rs
Br or I v
R3 R4
where L is a leaving group, for example halo such as fluoro.
The reaction of aniline and the benzoic acid derivative generally is
accomplished by mixing the benzoic acid with an equimolar quantity or excess
of the aniline in an unreactive organic solvent such as tetrahydrofuran or
toluene, in the presence of a base such as lithium diisopropylamide, n-butyl
lithium, sodium hydride, triethylamine, and Hunig's base. The reaction
generally is carried out at a temperature of about -78°C to about
100°C, and
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normally is complete within about 2 hours to about 4 days. The product can be
isolated by removing the solvent, for example by evaporation under reduced
pressure, and further purified, if desired, by standard methods such as
chromatography, crystallization, or distillation.
The 2-(phenylamino)-benzoic acid (eg, Formula IA, where R7 is
hydrogen) can be reacted with an organic or inorganic base such as pyridine,
triethylamine, calcium carbonate, or sodium hydroxide to produce a
pharmaceutically acceptable salt. The free acids can also be reacted with an
alcohol of the formula HORS (where R7 is other than hydrogen, for example
methyl) to produce the corresponding ester. Reaction of the benzoic acid with
an alcohol can be carried out in the presence of a coupling agent. Typical
coupling reagents include 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline
(EEDQ), 1,3-dicyclohexylcarbodiimide (DCC), bromo-tris(pyrrolidino)-
phosphonium hexafluorophosphate (PyBrOP), and (benzotriazolyloxy)
tripyrrolidino phosphonium hexafluorophosphate (PyBOP). The phenylamino
benzoic acid and alcohol derivative normally are mixed in approximately
equimolar quantities in an unreactive organic solvent such as
dichloromethane, tetrahydrofuran, chloroform, or xylene, and an equimolar
quantity of the coupling reagent is added. A base such as triethylamine or
diisopropylethylamine can be added to act as an acid scavenger if desired.
The coupling reaction generally is complete after about 10 minutes to 2 hours,
and the product is readily isolated by removing the reaction solvent, for
instance by evaporation under reduced pressure, and purifying the product by
standard methods such as chromatography or crystallizations from solvents
such as acetone, diethyl ether, or ethanol.
The benzamides of the invention, Formula I(A) where Z is CONRgR7,
are readily prepared by reacting the foregoing benzoic acids with an amine of
the formula HNRgR7. The reaction is carried out by reacting approximately
equimolar quantities of the benzoic acid and amine in an unreactive organic
solvent in the presence of a coupling reagent. Typical solvents are
chloroform,
dichloromethane, tetrahydrofuran, benzene, toluene, and xylene. Typical
coupling reagents include DCC, EEDQ, PyBrOP, and PyBOP. The reaction is
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generally complete after about 10 minutes to about 2 hours when carried out
at a temperature of about 0°C to about 60°C. The product amide
is readily
isolated by removing the reaction solvent, for instance by evaporation, and
further purification can be accomplished by normal methods such as
chromatography, crystallization, or distillation. The hydrazides
(z = CONHNR1pR11) are similarly prepared by coupling a benzoic acid with a
hydrazine of the formula H2HNR1pR11
The benzyl alcohols of the invention, compounds of Formula I(A) where
Z is CH20Rg and R6 is hydrogen, are readily prepared by reduction of the
corresponding benzoic acid according to the following scheme
0
R ~2 C - OH R ~2 HZOH
I ~ N ~ reducing I ~ N
RS agent I / I RS
Br or I Br or I
R3 R4 R3 R4
Typical reducing agents commonly employed include borane in
tetrahydrofuran. The reduction normally is carried out in an unreactive
organic
solvent such as tetrahydrofuran, and generally is complete within about
2 hours to about 24 hours when conducted at a temperature of about 0°C
to
about 40°C.
The following detailed examples 1A to 224A illustrate specific
compounds provided by this invention.
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D. Uses
The disclosed compositions are useful as both prophylactic and
therapeutic treatments for diseases or conditions relating to chronic pain,
including neuropathic pain, as provided in the Summary section, as well as
diseases or conditions modulated by the MEK cascade. For example, in one
embodiment, the disclosed method relates to postoperative pain, phantom
limb pain, burn pain, gout, trigeminal neuralgia, acute herpetic and
postherpetic pain, causalgia, diabetic neuropathy, plexus avulsion, neuroma,
vasculitis, crush injury, constriction injury, tissue injury, post-surgical
pain,
arthritis pain, or limb amputation
For example, local injuries can be treated with local or topical
administration. Chronic pain affecting the entire body, such as diabetic
neuropathy can be treated with systemic administration (injection or orally)
of
a disclosed composition. Treatment for chronic pain (e.g., post-operative
pain) confined to the lower body can be administered centrally, e.g.,
epidurally. Formulations and methods of administration can include the use of
more than one MEK inhibitor, or a combination of a MEK inhibitor and another
pharmaceutical agent, such as an anti-inflammatory, analgesic, muscle
relaxing, or anti-infective agent. Preferred routes of administration are
oral,
intrathecal or epidural, subcutaneous, intravenous, intramuscular, and, for
non-human mammals, intraplantar, and are preferably epidural.
1. Dosages
Those skilled in the art will be able to determine, according to known
methods, the appropriate dosage for a patient, taking into account factors
such as age, weight, general health, the type of pain requiring treatment, and
the presence of other medications. In general, an effective amount will be
between 0.1 and 1000 mg/kg per day, preferably between 1 and 300 mg/kg
body weight, and daily dosages will be between 10 and 5000 mg for an adult
subject of normal weight. Commercially available capsules or other
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formulations (such as liquids and film-coated tablets) of 100 mg, 200 mg, 300
mg, or 400 mg can be administered according to the disclosed methods.
2. Formulations
Dosage unit forms include tablets, capsules, pills, powders, granules,
S aqueous and nonaqueous oral solutions and suspensions, and parenteral
solutions packaged in containers adapted for subdivision into individual
doses.
Dosage unit forms can also be adapted for various methods of administration,
including controlled release formulations, such as subcutaneous implants.
Administration methods include oral, rectal, parenteral (intravenous,
intramuscular, subcutaneous), intracisternal, intravaginal, intraperitoneal,
intravesical, local (drops, powders, ointments, gels, or cream), and by
inhalation (a buccal or nasal spray).
Parenteral formulations include pharmaceutically acceptable aqueous
or nonaqueous solutions, dispersion, suspensions, emulsions, and sterile
powders for the preparation thereof. Examples of carriers include water,
ethanol, polyols (propylene glycol, polyethylene glycol), vegetable oils, and
injectable organic esters such as ethyl oleate. Fluidity can be maintained by
the use of a coating such as lecithin, a surfactant, or maintaining
appropriate
particle size. Carriers for solid dosage forms include (a) fillers or
extenders,
(b) binders, (c) humectants, (d) disintegrating agents, (e) solution
retarders, (f)
absorption acccelerators, (g) adsorbants, (h) lubricants, (i) buffering
agents,
and (j) propellants.
Compositions may also contain adjuvants such as preserving, wetting,
emulsifying, and dispensing agents; antimicrobial agents such as parabens,
chlorobutanol, phenol, and sorbic acid; isotonic agents such as a sugar or
sodium chloride; absorption-prolonging agents such as aluminum
monostearate and gelatin; and absorption-enhancing agents.
3. Related compounds
The invention provides the disclosed compounds and closely related,
pharmaceutically acceptable forms of the disclosed compounds, such as
salts, esters, amides, hydrates or solvated forms thereof; masked or protected
forms; and racemic mixtures, or enantiomerically or optically pure forms.
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Pharmaceutically acceptable salts, esters, and amides include
carboxylate salts (e.g., C ~_$ alkyl, cycloalkyl, aryl, heteroaryl, or non-
aromatic
heterocyclic), amino acid addition salts, esters, and amides which are within
a
reasonable benefit/risk ratio, pharmacologically effective, and suitable for
contact with the tissues of patients without undue toxicity, irritation, or
allergic
response. Representative salts include hydrobromide, hydrochloride, sulfate,
bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate,
laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate,
fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate,
lactiobionate, and laurylsulfonate. These may include alkali metal and alkali
earth cations such as sodium, potassium, calcium, and magnesium, as well as
non-toxic ammonium, quaternary ammonium, and amine cations such as
tetramethyl ammonium, methylamine, trimethylamine, and ethylamine. See,
for example, S.M. Berge, et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977,
66:1-19 which is incorporated herein by reference. Representative
pharmaceutically acceptable amides of the invention include those derived
from ammonia, primary C 1_6 alkyl amines and secondary di (C ~_6 alkyl)
amines. Secondary amines include 5- or 6-membered heterocyclic or
heteroaromatic ring moieties containing at least one nitrogen atom and
optionally between 1 and 2 additional heteroatoms. Preferred amides are
derived from ammonia, C ~_3 alkyl primary amines, and di (C ~_2 alkyl)amines.
Representative pharmaceutically acceptable esters of the invention include
C ~_~ alkyl, C 5_7 cycloalkyl, phenyl, and phenyl(C ~_6)alkyl esters.
Preferred
esters include methyl esters.
The invention also includes disclosed compounds having one or more
functional groups (e.g., hydroxyl, amino, or carboxyl) masked by a protecting
group. Some of these masked or protected compounds are pharmaceutically
acceptable; others will be useful as intermediates. Synthetic intermediates
and processes disclosed herein, and minor modifications thereof, are also
within the scope of the invention.
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HYDROXYL PROTECTING GROUPS
Hydroxyl protecting groups include: ethers, esters, and protection for 1,2-
and
1,3-diols. The ether protecting groups include: methyl, substituted methyl
ethers, substituted ethyl ethers, substituted benzyl ethers, silyl ethers and
conversion of silyl ethers to other functional groups.
Substituted Methyl Ethers
Substituted methyl ethers include: methoxymethyl, methylthiomethyl, t-
utylthiomethyl, (phenyldimethylsilyl) methoxymethyl, benzyloxymethyl, p-
ethoxybenzyloxymethyl, (4-methoxyphenoxy) methyl, guaiacolmethyl, t
butoxymethyl, 4-pentenyloxymethyl, siloxymethyl, 2-methoxyethoxymethyl,
2,2,2-trichloroethoxymethyl, bis(2-chloro- ethoxy)methyl, 2-
(trimethylsilyl)ethoxymethyl, tetrahydropyranyl, 3-bromotetrahydro-pyranyl,
tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl, 4-
methoxytetrahydrothio-pyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxido,
1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl, 1,4-dioxan-2-yl,
tetrahydrofuranyl, tetrahydrothiofuranyl, and 2,3,3a,4,5,6,7,7a-octahydro-
7,8,8-trimethyl-4,7-ethanobenzofuran-2-yl.
Substituted Ethyl Ethers
Substituted ethyl ethers include: 1-ethoxyethyl, 1-(2,chloroethoxy)ethyl,
1-methyl-1-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-
fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilyethyl, 2-
(phenylselenyl)ethyl, t-
butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, and benzyl.
Substituted Benzyl Ethers
Substituted benzyl ethers include: p-methoxybenzyl, 3,4-dimethoxybenzyl,
o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,
p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl,
p, p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, a-naphthyldiphenyl-
methyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,
tri-(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxy)phenyldiphenylmethyl,
4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-
tris(levulinoyloxyphenyl) methyl, 4,4',4"tris(benzoyloxyphenyl)methyl, 3-
(imidazol-1-ylmethyl)bis(4',4"-dimethoxyphenyl)-methyl, 1,1-bis(4-
methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl) xanthenyl, 9-(9-
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phenyl-10-oxo) anthryl, 1,3-benzodithiolan-2-yl, and benzisothiazolyl S,S-
dioxido.
Silyl Ethers
Silyl ethers include: trimethylsilyl, triethylsilyl, triisopropylsilyl,
dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl, t-
butyldimethylsilyl; t-butyldiphenylsilyl, tribenzylsilyl, tri-p-xylylsilyl,
triphenylsilyl,
diphenylmethylsilyl, and t-butylmethoxyphenylsilyl.
ESTERS
Esters protecting groups include: esters, carbonates, assisted cleavage,
miscellaneous esters, and sulfonates.
Esters
Examples of protective esters include: formate, benzoylformate, acetate,
chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate,
methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-
chlorophenoxyacetate, p-P-phenylacetate, 3-phenylpropionate, 4-
oxopentanoate (levulinate), 4,4-(ethylenedithio) pentanoate, pivaloate,
adamantoate,crotonate,4-methoxycrotonate, benzoate, p-phenylbenzoate,
and 2,4,6-trimethylbenzoate (mesitoate).
Carbonates
Carbonates include: methyl, 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl,
2-(trimethylsilyl) ethyl, 2-(phenylsulfonyl) ethyl, 2-(triphenylphosphonio)
ethyl,
isobutyl, vinyl, allyl, p-nitrophenyl, benzyl, p-methoxybenzyl, 3,4-
dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, S-benzyl thiocarbonate, 4-
ethoxy-1-naphthyl, and methyl dithiocarbonate.
Assisted Cleavage
Examples of assisted cleavage protecting groups include: 2-iodobenzoate, 4-
azido-butyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl) benzoate, 2-
formylbenzene-sulfonate, 2-(methylthiomethoxy) ethyl carbonate, 4-
(methylthiomethoxymethyl) benzoate, and 2-(methylthiomethoxymethyl)
benzoate.
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Miscellaneous Esters
In addition to the above classes, miscellaneous esters include: 2,6-dichloro-4-
methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)
phenoxyacetate, 2,4-bis(1,1-dimethylpropyl) phenoxyacetate,
chlorodiphenylacetate, isobutyrate, monosuccinoate, (E']-2-methyl-2-
butenoate (tigloate), o-(methoxycarbonyl) benzoate, p-P-benzoate, a-
naphthoate, nitrate, alkyl N,N,N' N'-tetramethylphosphorodiamidate, N-
phenylcarbamate, borate, dimethylphosphinothioyl, and 2,4-
dinitrophenylsulfenate.
Sulfonates
Protective sulfates includes: sulfate, methanesulfonate(mesylate),
benzylsulfonate, and tosylate.
PROTECTION FOR 1,2- AND 1,3-DIOLS
The protection for 1,2 and 1,3-diols group includes: cyclic acetals and
ketals,
cyclic ortho esters, and silyl derivatives.
Cyclic Acetals and Ketals
Cyclic acetals and ketals include: methylene, ethylidene, 1-t-butylethylidene,
1-phenylethylidene, (4-methoxyphenyl) ethylidene, 2,2,2-trichloroethylidene,
acetonide (isopropylidene), cyclopentylidene, cyclohexylidene,
cycloheptylidene, benzylidene, p-methoxybenzylidene, 2,4-
dimethoxybenzylidene, 3,4-dimethoxybenzylidene, and 2-nitrobenzylidene.
Cyclic Ortho Esters
Cyclic ortho esters include: methoxymethylene, ethoxymethylene, dimethoxy-
methylene, 1-methoxyethylidene, 1-ethoxyethylidine, 1,2-
dimethoxyethylidene, a-methoxybenzylidene, 1-(N,N-
dimethylamino)ethylidene derivative, a-(N,N-dimethylamino) benzylidene
derivative, and 2-oxacyclopentylidene.
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PROTECTION FOR THE CARBOXYL GROUP
ESTERS
Ester protecting groups include: esters, substituted methyl esters, 2-
substituted ethyl esters, substituted benzyl esters, silyl esters, activated
esters, miscellaneous derivatives, and stannyl esters.
Substituted Methyl Esters
Substituted methyl esters include: 9-fluorenylmethyl, methoxymethyl,
methylthiomethyl, tetrahydropyranyl, tetrahydrofuranyl, methoxyethoxymethyl,
2-(trimethylsilyl)ethoxy-methyl, benzyloxymethyl, phenacyl, p-bromophenacyl,
a-methylphenacyl, p-methoxyphenacyl, carboxamidomethyl, and N-
phthalimidomethyl.
2-Substituted Ethyl Esters
2-Substituted ethyl esters include: 2,2,2-trichloroethyl, 2-haloethyl, a-
chloroalkyl, 2-(trimethylsily)ethyl, 2-methylthioethyl, 1,3-dithianyl-2-
methyl,
2(p-nitrophenylsulfenyl)-ethyl, 2-(p-toluenesulfonyl)ethyl, 2-(2'-
pyridyl)ethyl, 2-
(diphenylphosphino)ethyl, 1-methyl-1-phenylethyl, t-butyl, cyclopentyl,
cyclohexyl, allyl, 3-buten-1-yl, 4-(trimethylsily)-2-buten-1-yl, cinnamyl, a-
methylcinnamyl, phenyl, p-(methylmercapto)-phenyl, and benzyl.
Substituted Benzyl Esters
Substituted benzyl esters include: triphenylmethyl, diphenylmethyl,
bis(o-nitrophenyl)methyl, 9-anthrylmethyl, 2-(9,10-dioxo)anthrylmethyl, 5-
dibenzo-suberyl, 1-pyrenylmethyl,2-(trifluoromethyl)-6-chromylmethyl, 2,4,6-
trimethylbenzyl, p-bromobenzyl, o=nitrobenzyl, p-nitrobenzyl, p-
methoxybenzyl, 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl, 4-sulfobenzyl,
piperonyl, and 4-P-benzyl.
Silyl Esters
Silyl esters include: trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, i-
propyldimethylsilyl, phenyldimethylsilyl, and di- t-butylmethylsilyl.
Miscellaneous Derivatives
Miscellaneous derivatives includes: oxazoles, 2-alkyl-1,3-oxazolines, 4-alkyl-
5-oxo-1,3-oxazolidines, 5-alkyl-4-oxo-1,3-dioxolanes, ortho esters, phenyl
group, and pentaaminocobalt(III) complex.
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Stannyl Esters
Examples of stannyl esters include: triethylstannyl and tri-n-butylstannyl.
AMIDES AND HYDRAZIDES
Amides include: N,N-dimethyl, pyrrolidinyl, piperidinyl, 5,6-
dihydrophenanthridinyl, o-nitroanilides, N-7-nitroindolyl, N-8-nitro-1,2,3,4-
tetrahydroquinolyl, and p-P-benzenesulfonamides. Hydrazides include: N-
phenyl, N,N'-diisopropyl and other dialkyl hydrazides.
PROTECTION FOR THE AMINO GROUP
CARBAMATES
Carbamates include: carbamates, substituted ethyl, assisted cleavage,
photolytic cleavage, urea-type derivatives, and miscellaneous carbamates.
Carbamates
Carbamates include: methyl and ethyl, 9-fluorenylmethyl, 9-(2-
sulfo)fluorenylmethyl, 9-(2,7-dibromo)fluorenylmethyl, 2,7-di-t butyl-[9-
(10,10-
dioxo-10,10,10,10-tetrahydro- thioxanthyl)]methyl, and 4-methoxyphenacyl.
Substituted Ethyl
Substituted ethyl protective groups include: 2,2,2-trichloroethyl, 2-
trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl, 1,1-
dimethyl-
2-haloethyl, 1,1 dimethyl-2,2-dibromoethyl, 1,1-dimethyl-2,2,2-trichloroethyl,
1-
methyl-1-(4-biphenylyl)ethyl, 1-(3,5-di-t-butylphenyl)-1-methylethyl, 2-(2'-
and
4'-pyridyl)ethyl, 2-(N,N-icyclohexylcarboxamido)- ethyl, t-butyl, 1-adamantyl,
vinyl, allyl, 1-isopropylallyl, connamyl, 4-nitrocinnamyl, quinolyl, N-
hydroxypiperidinyl, alkyldithio, benzyl, p-methoxybenzyl, p-nitrobenzyl, p-
bromobenzyl, p-chlorobenzyl, 2,4dichlorobenzyl, 4-methylsulfinylbenzyl,
9-anthrylmethyl, and diphenylmethyl.
Assisted Cleavage
Protection via assisted cleavage includes: 2-methylthioethyl,
2-methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl, [2-(1,3-dithianyl)]methyl,
4-methylthiophenyl, 2,4-dimethyl-thiophenyl, 2-phosphonioethyl,
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2-triphenylphosphonioisopropyl, 1,1-dimethyl-2cyanoethyl, m-chloro-p-
acyloxybenzyl, p-(dihydroxyboryl)benzyl, 5-benzisoxazolyl-methyl, and
2-(trifluoromethyl)-6-chromonylmethyl.
Photolytic Cleavage
Photolytic cleavage methods use groups such as: m-nitrophenyl, 3,5-
dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, and phenyl(o-
nitrophenyl)methyl.
Urea-Tyae Derivatives
Examples of of urea-type derivatives include: phenothiazinyl-(10)-carbonyl
derivative, N'-p-toluenesulfonylaminocarbonyl, and N'-
phenylaminothiocarbonyl.
Miscellaneous Carbamates
In addition to the above, miscellaneous carbamates include: t-amyl, S-benzyl
thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl, cyclopentyl,
cyclopropylmethyl, p-decyloxy-benzyl, diisopropylmethyl, 2,2-
dimethoxycarbonylvinyl, o-(N,N-dimethyl-carboxamido)-benzyl, 1,1-dimethyl
3(N,N-dimethylcarboxamido)propyl, 1,1-dimethyl-propynyl, di(2-pyridyl)methyl,
2-furanylmethyl, 2-iodoethyl, isobornyl, isobutyl, isonicotinyl, p(p'-
methoxyphenyl- azo)benzyl, 1-methylcyclobutyl, 1-methylcyclohexyl, 1-
methyl-1-cyclopropyl- methyl, 1-methyl-(3,5-dimethoxyphenyl)ethyl, 1-methyl-
1 (p-henylazophenyl)- ethyl, 1-methyl-1-phenylethyl, 1-methyl-1-(4-
pyridyl)ethyl, phenyl, p-(phenylazo)benzyl, 2,4,6-tri-t butylphenyl, 4-
(trimethylammonium) benzyl, and 2,4,6-trimethylbenzyl.
AMIDES
Amides
Amides includes: N-formyl, N-acetyl, N-chloroacetyl, N-trichloroacetyl,
N-trifluoroacetyl, N-phenylacetyl, N-3-phenylpropionyl, N-picolinoyl, N-3-
pyridyl-carboxamide, N-benzoylphenylalanyl derivative, N-benzoyl, and N-p-
phenylbenzoyl.
Assisted Cleavage
Assisted cleavage groups include: N-o-nitrophenylacetyl, N-o-
nitrophenoxyacetyl, N-acetoacetyl, (N'-dithiobenzyloxycarbonylamino)acetyl,
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N-3-(p-hydroxphenyl) propionyl, N-3-(o-nitrophenyl)propionyl, N-2-methyl-2-
(o-nitrophenoxy)propionyl, N-2-methyl-2-(o-phenylazophenoxy)propionyl, N-4-
chlorobutyryl, N-3-methyl-3-nitrobutyryl, N-o-nitrocinnamoyl, N-
acetylmethionine derivative, N-o-nitrobenzoyl, N-o-
(benzoyloxymethyl)benzoyl, and 4,5-diphenyl-3-oxazolin-2-one.
Cyclic Imide Derivatives
Cyclic imide derivatives include: N-phthalimide, N-dithiasuccinoyl,
N-2,3-diphenyl-maleoyl, N-2,5-dimethylpyrrolyl,
N-1,1,4,4-tetramethyldisilylazacyclopentane adduct, 5-substituted
1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-
1,3,5-triazacyclohexan-2-one, and 1-substituted 3,5-dinitro-4-pyridonyl.
SPECIAL -NH PROTECTIVE GROUPS
Protective groups for - NH include: N-alkyl and N-aryl amines, imine
derivatives, enamine derivatives, and N-hetero atom derivatives (such as N-
metal, N-N, N-P, N-Si, and N-S), N-sulfenyl, and N-sulfonyl.
N-Alkyl and N-Aryl Amines
N-alkyl and N-aryl amines include: N-methyl, N-allyl,
N-[2-(trimethylsilyl)ethoxyl]-methyl, N-3-acetoxypropyl,
N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), quaternary ammonium salts,
N-benzyl, N-di(4-methoxyphenyl)methyl, N-5-dibenzosuberyl,
N-triphenylmethyl, N-(4-methoxyphenyl)diphenylmethyl, N-9-phenylfluorenyl,
N-2,7-dichloro-9-fluorenylmethylene, N-ferrocenylmethyl, and
N-2-picolylamine N'-oxide.
Imine Derivatives
Imine derivatives include: N-1,1-dimethylthiomethylene, N-benzylidene,
N-p-methoxybenzylidene, N-diphenylmethylene,
N-[(2-pyridyl)mesityl]methylene, N-(N',N'-dimethylaminomethylene),
N,N'-isopropylidene, N-p-nitrobenzylidene, N-salicylidene,
N-5-chlorosalicylidene, N-(5-chloro-2-hydroxyphenyl)phenyl-methylene, and
N-cyclohexylidene.
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Enamine Derivative
An example of an enamine derivative is N-
(5,5-dimethyl-3-oxo-1-cyclohexenyl).
N-Hetero Atom Derivatives
N-metal derivatives include: N-borane derivatives, N-diphenylborinic acid
derivative, N-[phenyl(pentacarbonylchromium- or -tungsten)]carbenyl, and
N-copper or N-zinc chelate. Examples of N-N derivatives include: N-nitro,
N-nitroso, and N-oxide. Examples of N-P derivatives include:
N-diphenylphosphinyl, N-dimethylthiophosphinyl, N-diphenylthiophosphinyl,
N-dialkyl phosphoryl, N-dibenzyl phosphoryl, and N-diphenyl phosphoryl.
Examples of N-sulfenyl derivatives include: N-benzenesulfenyl,
N-o-nitrobenzenesulfenyl, N-2,4-dinitrobenzenesulfenyl,
N-pentachlorobenzenesulfenyl, N-2-nitro-4-methoxy-benzenesulfenyl,
N-triphenylmethylsulfenyl, and N-3-nitropyridinesulfenyl. N-sulfonyl
derivatives include: N-p-toluenesulfonyl, N-benzenesulfonyl,
N-2,3,6-trimethyl- 4-methoxybenzenesulfonyl,
N-2,4,6-trimethoxybenzenesulfonyl, N-2,6-dimethyl-4-methoxy-
benzenesulfonyl, N-pentamethylbenzenesulfonyl,
N-2,3,5,6-tetramethyl-4-methoxybenzene- sulfonyl,
N-4-methoxybenzenesulfonyl, N-2,4,6-trimethylbenzenesulfonyl,
N-2,6-dimethoxy- 4-methylbenzenesulfonyl,
N-2,2,5,7,8-pentamethylchroman-6-sulfonyl, N-methanesulfonyl,
N-~trimethylsilylethanesulfonyl, N-9-anthracenesulfonyl,
N-4-(4',8'-dimethoxynaphthylmethyl)-benzenesulfonyl, N-benzylsulfonyl,
N-trifluoromethylsulfonyl, and N-phenacylsulfonyl.
Disclosed compounds which are masked or protected may be prodrugs,
compounds metabolized or otherwise transformed in vivo to yield a disclosed
compound, e.g., transiently during metabolism. This transformation may be a
hydrolysis or oxidation which results from contact with a bodily fluid such as
blood, or
the action of acids, or liver, gastrointestinal, or other enzymes.
Features of the invention are further described in the examples below.
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E. Examples
BIOLOGICAL EXAMPLES
EXAMPLE 1
Effect of PD 198306 on streptozocin-induced static allodynia
Animals
Male Sprague Dawley rats (250-300g), obtained from Bantin and
Kingman, (Hull, U.K.) were housed in groups of 3. All animals were kept under
a 12h light/dark cycle (lights on at 07h OOmin) with food and water ad
libitum.
All experiments were carried out by an observer blind to drug treatments.
Development of diabetes in the rat
Diabetes was induced in rats by a single i.p. injection of streptozocin
(50 mg/kg) as described previously (Courteix et al., 1993).
Evaluation of static allodynia
Mechanical hypersensitivity was measured using Semmes-Weinstein
von Frey hairs (Stoelting, Illinois, U.S.A.). Animals were placed into wire
mesh
bottom cages allowing access to the underside of their paws. Animals were
habituated to this environment prior to the start of the experiment.
Mechanical
hypersensitivity was tested by touching the plantar surface of the animals
right
hind paw with von Frey hairs in ascending order of force ( 0.7, 1.2, 1.5, 2,
3.6,
5.5, 8.5, 11.8, 15.1 and 29g) for up to 6 sec. Once a withdrawal response was
established, the paw was re-tested, starting with the next descending von
Frey hair until no response occurred. The highest force of 29 g lifted the paw
as well as eliciting a response, thus represented the cut off point. The
lowest
amount of force required to elicit a response was recorded as the paw
withdrawal threshold (PWT) in grams.
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Drugs
PD 198306 [N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-
phenylamino)-benzamide] and CI-1008 (pregabalin) were synthesized at
Parke-Davis (Ann Arbor, MI, USA). PD 198306 was suspended in
cremophor:ethanol:water (1:1:8) vehicle. Pregabalin was dissolved in water.
Both compounds were administered orally. Streptozocin (Aldrich, UK) was
dissolved in 0.9% w/v NaCI and administered intraperitoneally. Drug
administrations were made in a volume of 1 ml/kg.
Statistics
The static allodynia data were analysed using a Kruskall-Wallis ANOVA
for non-parametric results, followed when significant by Mann-Whitney's t
test.
Experimental protocol
Static allodynia was assessed with von Frey hairs, before (baseline,
BL) and 1 h after oral administration of PD 198306 (30mg/kg, p.o.), vehicle
(cremophor:ethanol:water, 1:1:8) or pregabalin (30mg/kg, p.o.) (test).
Animals were administered again the same compounds on the following day,
both in the morning and the afternoon. Static allodynia was assessed only
before and 1 h after the afternoon administration, in order to minimise the
habituation of the animals to the testing conditions. Animals treated with
pregabalin received water in the morning administration, in order to avoid the
potential development of tolerance to the compound with repeated
administration.
Day 1: Day 2:
a.m.: PD 198306
Water
Vehicle
p.m.: BL p.m.: BL
PD 198306 PD 198306
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Pregabalin Pregabalin
Vehicle Vehicle
Test Test
RESULTS
A single administration of pregabalin (30mg/kg, p.o.) significantly
blocked streptozocin-induced static allodynia 1 h after administration. In
contrast, a single administration of PD 198306 (30mg/kg, p.o) had no effect on
streptozocin-induced static allodynia 1 h after administration (see below).
However, after the compound had been administered twice more on the
following day, it significantly blocked streptozocin-induced static allodynia
1 h
after the third administration. The effects had disappeared by the following
day (see FIG. 1 ).
EXAMPLE 2
MATERIALS AND METHODS
Animals
Male Sprague Dawley rats (250-300g), obtained from Charles River,
Margate, U.K.) were housed in groups of 3-6. All animals were kept under a
12h light/dark cycle (lights on at 07h OOmin) with food and water ad libitum.
All
experiments were carried out by an observer blind to drug treatments.
Diabetes was induced in rats by a single i.p. injection of streptozocin
(50mg/kg) as described previously (Courteix et al., 1993).
Development of Chronic Constriction Iniury in the rat
Animals were anaesthetised with 2% isoflurane 1:4 02/N20 mixture
maintained during surgery via a nose cone. The sciatic nerve was ligated as
previously described by Bennett and Xie, 1988. Animals were placed on a
homeothermic blanket for the duration of the procedure. After surgical
preparation the common sciatic nerve was exposed at the middle of the thigh
by blunt dissection through biceps femoris. Proximal to the sciatic
trifurcation,
about 7mm of nerve was freed of adhering tissue and 4 ligatures (4-0 silk)
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were tied loosely around it with about 1 mm spacing. The incision was closed
in layers and the wound treated with topical antibiotics.
Intrathecal infections
PD 198306 and pregabalin were administered intrathecally in a volume
of 10 ~I using a 100 ~I Hamilton syringe by exposing the spine of the rats
under
brief isoflurane anaesthesia. Injections were made into the intrathecal space
between lumbar region 5-6 with a 10 mm long 27 gauge needle. Penetrations
were judged successful if there was a tail flick response. The wound was
sealed
with an autoclip and rats appeared fully awake within 2-3 min following
injection.
Evaluation of static allodynia
Mechanical hypersensitivity was measured using Semmes-Weinstein
von Frey hairs (Stoelting, Illinois, U.S.A.). Animals were placed into wire
mesh
bottom cages allowing access to the underside of their paws. Animals were
habituated to this environment prior to the start of the experiment.
Mechanical
hypersensitivity was tested by touching the plantar surface of the animals
right
hind paw with von Frey hairs in ascending order of force ( 0.7, 1.2, 1.5, 2,
3.6,
5.5, 8.5, 11.8, 15.1 and 29g) for up to 6sec. Once a withdrawal response was
established, the paw was re-tested, starting with the next descending von
Frey hair until no response occurred. The highest force of 29g lifted the paw
as well as eliciting a response, thus represented the cut off point. The
lowest
amount of force required to elicit a response was recorded as the paw
withdrawal threshold (PWT) in grams.
Experimental protocol
Static allodynia was assessed with von Frey hairs, before (baseline,
BL) and 0.5h, 1 h and 2h after intrathecal or intraplantar administration of
PD
198306 (1-30~g, i.t.), vehicle (cremophor:ethanol:water, 1:1:8) or pregabalin
(10pg, i.t). For oral administration experiments, static allodynia was
assessed
with von Frey hairs, before (baseline, BL) and 1h after oral administration of
PD 198306 (3-30mg/kg, p.o.), vehicle (cremophor:ethanol:water, 1:1:8) or
pregabalin (30mg/kg, p.o.). Animals were administered again the same
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compounds on the following day, both in the morning and the afternoon. Static
allodynia was assessed before and 1 h after the morning administration. In the
afternoon static allodynia was assessed before, 1 h, 2h and 3h after
administration for streptozocin treated animals. CCI animals were assessed
before, 1 h and 2h after administration
Drugs used
PD 198306 and pregabalin were synthesised at Parke-Davis (Ann
Arbor, MI, USA). PD 198306 was suspended in cremophor:ethanol:water
(1:1:8) vehicle. Pregabalin was dissolved in water. Both compounds were
administered orally, intrathecally or intraplantar in volumes of 1 ml/kg, 1
Op,l and
100p1 respectively. Streptozocin (Aldrich, UK) was dissolved in 0.9% w/v NaCI
and administered intraperitoneally in a volume of 1 ml/kg.
Statistics
Data were analysed using a Kruskall-Wallis ANOVA for non-parametric
results, followed when significant by Mann-Whitney's t test vs vehicle group.
RESULTS
1. Effects of PD 198306 on static allodynia, following systemic
administration
1.1. Effect of PD198306 on streptozocin-induced static allodynia
A single administration of pregabalin (30mg/kg, p.o.) significantly blocked
streptozocin-induced static allodynia 1 h after administration. In contrast, a
single administration of PD 198306 (3-30mg/kg, p.o) had no effect on
streptozocin-induced static allodynia 1 h after administration (FIG. 2).
However, after the compound had been administered twice more on the
following day, PD 198306 (30mg/kg) significantly blocked streptozocin-
induced static allodynia for 2h after the third administration (FIG. 2).
1.2. Effect of PD198306 on CCI-induced static allodynia
A single administration of pregabalin (30mg/kg, p.o.) significantly blocked
CCI-
induced static allodynia 1 h after administration. In contrast, neither a
single or
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multiple administration of PD 198306 (3-30mg/kg, p.o) had any effect on CCI-
induced static allodynia (FIG. 3).
2. Effects of PD 198306 on static allodynia, following intrathecal
administration
Intrathecally administered PD198306 (1-30~,g) dose-dependently blocked the
maintenance of static allodynia in both streptozocin (FIG. 4) and CCI animals
(FIG. 5) with respective MEDs of 3 and 10 fig. This antiallodynic effect
lasted
for 1 h.
3. Effects of PD 198306 on static allodynia, following intraplantar
administration
An intrathecal administration of PD 198306 (30~g) significantly blocked static
allodynia in both neuropathic pain models (FIGS. 6,7). In contrast, a single
administration of PD 198306 at a dose 100-fold higher (3mg/100~1) directly
into the paw had no effect on streptozocin (FIG. 6) or CCI-induced static
allodynia (FIG. 7).
REFERENCES
Bennett GJ, Xie Y-K. A peripheral mononeuropathy in rat that produces
disorders of pain sensation like those seen in man. Pain 1988;33:87-107.
Courteix C, Eschalier A and Lavarenne J. Streptozocin -induced rats:
behavioural evidence for a model of chronic pain. Pain 1993;53:81-8
EXAMPLE 3
Effect of other MEK inhibitors in a neuropathic pain model in the rat
SUMMARY
The effect of several MEK inhibitors, with different binding affinities, has
been investigated in the CCI model of neuropathic pain in the rat, by
assessing
static allodynia with von Frey hairs. Intrathecal administration of PD219622
or
PD297447 (30~g) had no significant effect on allodynia. This lack of effect
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may reflect the low affinity or solubility of the compounds. However,
intrathecal administration of PD 254552 or PD 184352 (30~g), which posses
higher binding affinities, blocked the maintenance of static allodynia in CCI
animals. The antiallodynic effect was only evident for 30min post-injection
and
thus, shorter than the one observed for pregabalin (100~g). The magnitude of
the effect was similar for 30~,g of PD 184352 and 100pg of pregabalin. From
this study it is concluded that MEK inhibitors exert an antiallodynic effect
in
CCI-induced neuropathic rats when administered intrathecally, and that the
antiallodynic effect correlates with the affinity of the compounds.
The animals and methods for developing chronic constriction injury in
the rat, injecting test compounds, and evaluation of static allodynia were
according to Example 2 above. PD219622, PD297447, PD 184352, PD
254552 and pregabalin were administered intrathecally at doses of 30 g for all
PD compounds and 100p,g for pregabalin. Static allodynia was assessed with
von Frey hairs, before (baseline, BL) and 0.5h, 1 h and 2h after intrathecal
administration of the compounds
Drugs used
PD297447, PD219622, PD 254552, PD 184352 (CI-1040), and pregabalin
were synthesised at Parke-Davis (Ann Arbor, MI, USA). PD297447,
PD219622, PD 254552 and PD 184352 were suspended in
cremophor:ethanol:water (1:1:8) vehicle. Pregabalin was dissolved in water.
All
compounds were administered intrathecally in a 10p,1 volume.
Statistics
Data were analysed using a Kruskall-Wallis ANOVA for non-parametric
results, followed when significant by Mann-Whitney's t test vs vehicle group.
RESULTS
Intrathecally administered PD297447 or PD219622 (30wg) had no
significant effect on allodynia. This lack of effect may reflect the low
affinity of
the compounds (965nM and 100nM respectively). However, intrathecal
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administration of PD 184352 or PD 254552 (30~g) blocked the maintenance
of static allodynia in CCI animals (see FIG. 8). These compounds possess
higher affinity (2 and 5 nM respectively). The antiallodynic effect was only
evident for 30min post-injection and thus, shorter than the one observed for
pregabalin (100p,g). The magnitude of the effect was similar for 30~g of PD
184352 and 100~.g of pregabalin.
The results indicate that MEK inhibitors exert an antiallodynic effect in
CCI-induced neuropathic rats when administered intrathecally, and that the
antiallodynic effect correlates with the affinity of the compounds.
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CHEMICAL EXAMPLES
EXAMPLE 1
4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide
(a) Preparation of 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic
acid
To a stirred solution containing 3.16 g (0.0133 mol) of 2-amino-
5-iodotoluene in 5 mL of tetrahydrofuran at -78°C was added 10 mL
(0.020 mol) of a 2.0 M lithium diisopropylamide in
tetrahydrofuran/heptane/ethylbenzene (Aldrich) solution. The resulting green
suspension was stirred vigorously for 15 minutes, after which time a solution
of 1.00 g (0.00632 mol) of 2,4-difluorobenzoic acid in 10 mL of
tetrahydrofuran
was added. The reaction temperature was allowed to increase slowly to room
temperature, at which temperature the mixture was stirred for 2 days. The
reaction mixture was concentrated by evaporation of the solvent under
reduced pressure. Aqueous HCI (10%) was added to the concentrate, and the
solution was extracted with dichloromethane. The organic phase was dried
(MgS04) and then concentrated over a steambath to low volume (10 mL) and
cooled to room temperature. The off-white fibers which formed were collected
by vacuum filtration, rinsed with hexane, and dried in a vacuum-oven
(76°C;
ca. 10 mm of Hg) to afford 1.10 g (47%) of the desired material;
mp 224-229.5°C;
1 H NMR (400 MHz, DMSO): ~ 9.72 (s, 1 H), 7.97 (dd, 1 H, J=7.0, 8.7 Hz),
7.70 (d, 1 H, J=1.5 Hz), 7.57 (dd, 1 H, J=8.4, 1.9 Hz), 7.17 (d, 1 H, J=8.2
Hz),
6.61-6.53 (m, 2H), 2.18 (s, 3H);
13C NMR (100 MHz, DMSO): b 169.87, 166.36 (d, JC_F=249.4 Hz),
150.11 (d, JC_F=11.4 Hz), 139.83, 138.49, 136.07, 135.26 (d, JC_F=11.5 Hz),
135.07, 125.60, 109.32, 104.98 (d, JC_F=21.1 Hz), 99.54 (d, JC-F=26.0 Hz),
89.43, 17.52;
19F NMR (376 MHz, DMSO): b -104.00 to -104.07 (m);
IR (KBr) 1670 (C=O stretch)cm-1;
MS (CI) M+1 = 372.
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Analysis calculated for C14H11 FIN02:
C, 45.31; H, 2.99; N, 3.77.
Found: C, 45.21; H, 2.77; N, 3.64.
(b) Preparation of 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-
phenylamino)-benzamide
To a stirred solution of 4-fluoro-2-(4-iodo-2-methyl-phenylamino)-
benzoic acid (0.6495 g, 0.001750 mol), O-(tetrahydro-2H-pyran-2-yl)-
hydroxylamine (0.2590 g, 0.002211 mol), and diisopropylethylamine (0.40 mL,
0.0023 mol) in 31 mL of an equivolume tetrahydrofuran-dichloromethane
solution was added 1.18 g (0.00227 mol) of solid PyBOP
([benzotriazolyloxy]tripyrrolidino phosphonium hexafluorophosphate,
Advanced ChemTech) directly. The reaction mixture was stirred for
30 minutes after which time it was concentrated in vacuo. The brown oil was
treated with 10% aqueous hydrochloric acid. The suspension was extracted
with ether. The organic extraction was washed with 10% sodium hydroxide
followed by another 10% hydrochloric acid wash, was dried (MgS04) and
concentrated in vacuo to afford 1.0 g of a light-brown foam. This intermediate
was dissolved in 25 mL of ethanolic hydrogen chloride, and the solution was
allowed to stand at room temperature for 15 minutes. The reaction mixture
was concentrated in vacuo to a brown oil that was purified by flash silica
chromatography. Elution with dichloromethane -~ dichloromethane-methanol
(166:1) afforded 0.2284 g of a light-brown viscous oil. Scratching with
pentane-hexanes and drying under high vacuum afforded 0.1541 g (23%) of
an off-white foam; mp 61-75°C;
1 H NMR (400 MHz, DMSO): b 11.34 (s, 1 H), 9.68 (s, 1 H), 9.18 (s, 1 H),
7.65 (d, 1 H, J=1.5 Hz), 7.58 (dd, 1 H, J=8.7, 6.8 Hz), 7.52 (dd, 1 H, J=8.4,
1.9 Hz), 7.15 (d, 1 H, J=8.4 Hz), 6.74 (dd, 1 H, J=11.8, 2.4 Hz), 6.62 (ddd, 1
H,
J=8.4, 8.4, 2.7 Hz), 2.18 (s, 3H);
13C NMR (100 MHz, DMSO): b 165.91, 164.36 (d, JC_F=247.1 Hz), 146.78,
139.18, 138.77, 135.43, 132.64, 130.60 (d, JC_F=11.5 Hz), 122.23, 112.52,
104.72 (d, J=22.1 Hz), 100.45 (d, JC_F=25.2 Hz), 86.77, 17.03;
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19F NMR (376 MHz, DMSO): b -107.20 to -107.27 (m);
IR (KBr) 3307 (broad, O-H stretch), 1636 (C=O stretch) cm-1;
MS (CI) M+1 = 387.
Analysis calculated for C14H12FIN202:
C, 43.54; H, 3.13; N, 7.25.
Found: C, 43.62; H, 3.24; N, 6.98.
EXAMPLE 2
5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-
benzamide
(a) Preparation of 5-Bromo-2,3,4-trifluorobenzoic acid
To a stirred solution comprised of 1-bromo-2,3,4-trifluorobenzene
(Aldrich, 99%; 5.30 g, 0.0249 mol) in 95 mL of anhydrous tetrahydrofuran
cooled to -78°C was slowly added 12.5 mL of 2.0 M lithium
diisopropylamide
in heptane/tetrahydrofuran/ethylbenzene solution (Aldrich). The mixture was
stirred for 1 hour and transferred by canula into 700 mL of a stirred
saturated
ethereal carbon dioxide solution cooled to -78°C. The cold bath was
removed,
and the reaction mixture was stirred for 18 hours at ambient temperature.
Dilute (10%) aqueous hydrochloric acid (ca. 500 mL) was poured into the
reaction mixture, and the mixture was subsequently concentrated on a rotary
evaporator to a crude solid. The solid product was partitioned between diethyl
ether (150 mL) and aq. NCI (330 mL, pH 0). The aqueous phase was
extracted with a second portion (100 mL) of diethyl ether, and the combined
ethereal extracts were washed with 5% aqueous sodium hydroxide (200 mL)
and water (100 mL, pH 12). These combined alkaline aqueous extractions
were acidified to pH 0 with concentrated aqueous hydrochloric acid. The
resulting suspension was extracted with ether (2 x 200 mL). The combined
organic extracts were dried (MgS04), concentrated in vacuo, and subjected to
high vacuum until constant mass was achieved to afford 5.60 g (88% yield) of
an off-white powder; mp 139-142.5°C;
1 H NMR (400 MHz, DMSO): b 13.97 (broad s, 1 H, 8.00-7.96 (m, 1 H);
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13C NMR (100 MHz, DMSO): b 162.96, 129.34, 118.47, 104.54 (d,
JC_F=22.9 Hz);
19F NMR (376 MHz, DMSO): b -120.20 to -120.31 (m), -131.75 to
-131.86 (m), -154.95 to -155.07 (m);
IR (KBr) 1696 (C=O stretch)cm-1;
MS (CI) M+1 = 255.
Analysis calculated for C74H21 BrF302:
C, 32.97; H, 0.79; N, 0.00; Br, 31.34; F, 22.35.
Found: C, 33.18; H, 0.64; N, 0.01; Br, 30.14; F, 22.75.
(b) Preparation of 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-
phenylamino)-benzoic acid
To a stirred solution comprised of 1.88 g (0.00791 mol) of 2-amino-
5-iodotoluene in 10 mL of tetrahydrofuran at -78°C was added 6 mL
(0.012 mol) of a 2.0 M lithium diisopropylamide in
tetrahydrofuran/heptane/ethylbenzene (Aldrich) solution. The resulting green
suspension was stirred vigorously for 10 minutes, after which time a solution
of 1.00 g (0.00392 mol) of 5-bromo-2,3,4-trifluorobenzoic acid in 15 mL of
tetrahydrofuran was added. The cold bath was subsequently removed, and
the reaction mixture stirred for 18 hours. The mixture was concentrated, and
the concentrate was treated with 100 mL of dilute (10%) aqueous hydrochloric
acid. The resulting suspension was extracted with ether (2 x 150 mL), and the
combined organic extractions were dried (MgS04) and concentrated in vacuo
to give an orange solid. The solid was triturated with boiling
dichloromethane,
cooled to ambient temperature, and collected by filtration. The solid was
rinsed with dichloromethane, and dried in the vacuum-oven (80°C) to
afford
1.39 g (76%) of a yellow-green powder; mp 259.5-262°C;
1 H NMR (400 MHz, DMSO): b 9.03 (s, 1 H), 7.99 (dd, 1 H, J=7.5, 1.9 Hz),
7.57 (dd, 1 H, J=1.5 Hz), 7.42 (dd, 1 H, J=8.4, 1.9 Hz), 6.70 (dd, 1 H, J=8.4,
6.0 Hz), 2.24 (s, 3H);
19F NMR (376 MHz, DMSO): b -123.40 to -123.47 (m); -139.00 to
-139.14 (m);
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IR (KBr) 1667 (C=O stretch)cm-1;
MS (CI) M+1 = 469.
Analysis calculated for Cl4HgBrF21N02:
C, 35.93; H, 1.94; N, 2.99; Br, 17.07; F, 8.12; I, 27.11.
Found: C, 36.15; H, 1.91; N, 2.70; Br, 16.40; F, 8.46; I, 26.05.
(c) Preparation of 5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-
phenylamino)-benzamide
To a stirred solution comprised of 5-bromo-3,4-difluoro-2-(4-iodo-
2-methyl-phenylamino)-benzoic acid (0.51 g, 0.0011 mol), O-(tetrahydro-2H-
pyran-2-yl)-hydroxylamine (0.15 g, 0.0013 mol), and diisopropylethylamine
(0.25 mL, 0.0014 mol) in 20 mL of an equivolume tetrahydrofuran-
dichloromethane solution was added 0.6794 g (0.001306 mol) of solid PyBOP
(Advanced ChemTech) directly. The reaction mixture was stirred at 24°C
for
10 minutes, and then was concentrated to dryness in vacuo. The concentrate
was suspended in 100 mL of 10% aqueous hydrochloric acid. The suspension
was extracted with 125 mL of diethyl ether. The ether layer was separated,
washed with 75 mL of 10% aqueous sodium hydroxide, and then with 100 mL
of dilute acid. The ether solution was dried (MgS04) and concentrated
in vacuo to afford 0.62 g (100%) of an off-white foam. The foam was dissolved
in ca. 15 mL of methanolic hydrogen chloride. After 5 minutes, the solution
was concentrated in vacuo to an oil, and the oil was purified by flash silica
chromatography. Elution with dichloromethane -~ dichloromethane-methanol
(99:1) afforded 0.2233 g (42%) of a yellow powder. The powder was dissolved
in diethyl ether and washed with dilute hydrochloric acid. The organic phase
was dried (MgS04) and concentrated in vacuo to afford 0.200 g of a foam.
This product was triturated with pentane to afford 0.1525 g of a powder that
was repurified by flash silica chromatography. Elution with dichloromethane
afforded 0.0783 g (15%) of an analytically pure title compound, mp 80-
90°C;
1 H NMR (400 MHz, DMSO): b 11.53 (s, 1 H), 9.38 (s, 1 H), 8.82 (s, 1 H),
7.70 (dd, 1 H, J=7.0, 1.9 Hz), 7.53 (s, 1 H), 7.37 (dd, 1 H, J=8.4, 1.9 Hz),
6.55 (dd, 1 H, J=8.2, 6.5 Hz), 2.22 (s, 3H);
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19F NMR (376 MHz, DMSO): b -126.24 to -126.29 (m), -137.71 to
-137.77 (m);
IR (KBr) 3346 (broad, O-H stretch), 1651 (C=O stretch)cm-1;
MS (CI) M+1 = 484.
Analysis calculated for C14H10BrF21N202:
C, 34.81; H, 2.09; N, 5.80.
Found: C, 34.53; H, 1.73; N, 5.52,
Examples 3 to 12 and 78 to 102 in the table below were prepared by
the general procedures of Examples 1 and 2.
EXAMPLES 13-77
Examples 13 to 77 were prepared utilizing combinatorial synthetic
methodology by reacting appropriately substituted phenylamino benzoic acids
R6
(e.g., as shown in Scheme 1) and hydroxylamines (e.g., HN-O-R7). A general
method is given below:
To a 0.8 mL autosampler vial in a metal block was added 40 pL of a
0.5 M solution of the acid in DMF and 40 pL of the hydroxylamine (2 M
solution in Hunig's base and 1 M in amine in DMF). A 0.5 M solution of
PyBrop was freshly prepared, and 50 pL were added to the autosampler vial.
The reaction was allowed to stand for 24 hours.
The reaction mixture was transferred to a 2 dram vial and diluted with
2 mL of ethyl acetate. The organic layer was washed with 3 mL of distilled
water and the water layer washed again with 2 mL of ethyl acetate. The
combined organic layers were allowed to evaporate to dryness in an open
fume hood.
The residue was taken up in 2 mL of 50% acetonitrile in water and
injected on a semi-prep reversed phase column (10 mm x 25 cm, 5 ~M
spherical silica, pore Size 115 A derivatised with C-18, the sample was eluted
at 4.7 mUmin with a linear ramp to 100% acetonitrile over 8.5 minutes. Elution
with 100% acetonitrile continued for 8 minutes.) Fractions were collected by
monitoring at 214 nM. The desired fractions were evaporated using a Zymark
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Turbovap. The product was dissolved in chloroform and transferred to a
preweighed vial, evaporated, and weighed again to determine the yield. The
structure was confirmed by mass spectroscopy.
EXAMPLES 3-102
Example Compound Melting MS
No. Point (°C) (M-H+)
3 2-(4-bromo-2-methyl-phenylamino)-4-fluoro- 56-75 dec 523
N-hydroxy-benzamide
4 5-Chloro-N-hydroxy-2-(4-iodo-2-methyl- 65 dec
phenylamino)-benzamide
5-Chloro-N-hydroxy-2-(4-iodo-2-methyl- 62-67
phenylamino)-N-methyl-benzamide
6 5-Chloro-2-(4-iodo-2-methyl-phenylamino)- 105-108
N-(terahydropyran-2-yloxy)benzamide
7 5-Chloro-2-(4-iodo-2-methyl-phenylamino)- 64-68
N-methoxybenzamide
8 4-Fluoro-N-hydroxy-2-(4-fluoro-2-methyl- 119-135
phenylamino)-benzamide
9 4-Fluoro-N-hydroxy-2-(2-methyl 101-103
phenylamino)-benzamide
4-Fluoro-2-(4-fluor-2-methyl-phenylamino)- 142-146
N-(terahydropyran-2-yloxy)benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
11 4-Fluoro-N-hydroxy-2-(4-cluoro-2-methyl- 133.5-135
phenylamino)-benzamide
12 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 107-109.5
N-phenylmethoxy-benzamide
13 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 399
N-methoxy-benzamide
14 3,4-Difluoro-2-(4-iodo-2-methyl- 417
phenylamino)-N-methoxy-benzamide
15 2-(4-Bromo-2-methyl-phenylamino)- 369
3,4-difluoro-N-methoxy-benzamide
16 2-(4-Bromo-2-methyl-phenylamino)-N- 342*
ethoxy-3,4-difluoro-benzamide (M-Et0)
17 5-Bromo-N-ethoxy-3,4-difluoro-2-(4-iodo- 509
2-methyl-phenylamino)-benzamide
18 3,4-Difluoro-2-(4-iodo-2-methyl- 445
phenylamino)-N-isopropoxy-benzamide
19 2-(4-Bromo-2-methyl-phenylamino)- 397
3,4-difluoro-N-isopropoxy-benzamide
20 4-Fluoro-N-(furan-3-ylmethoxy)-2-(4-iodo- 465
2-methyl-phenylamino)-benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
21 3,4-Difluoro-N-(furan-3-ylmethoxy)- 483
2-(4-iodo-2-methyl-phenylamino)-benzamide
22 2-(4-Bromo-2-methyl-phenylamino)- 435
3,4-difluoro-N-(furan-3-ylmethoxy)-
benzamide
23 5-Bromo-3,4-difluoro-N-(furan-3-ylmethoxy)- 561
2-(4-iodo-2-methyl-phenylamino)-benzamide
24 5-Bromo-N-(but-2-enyloxy)-3,4-difluoro- 536
2-(4-iodo-2-methyl-phenylamino)-benzamide
25 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 423
N-(prop-2-ynyloxy)-benzamide
26 3,4-Difluoro-2-(4-iodo-2-methyl- 441
phenylamino)-N-(prop-2-ynyloxy)-
benzamide
27 3,4-Difluoro-2-(4-iodo-2-methyl- 455
phenylamino)-N-(1-methyl-prop-2-ynyloxy)-
benzamide
28 2-(4-Bromo-2-methyl-phenylamino)- 407
3,4-difluoro-N-(1-methyl-prop-2-ynyloxy)-
benzamide
29 N-(But-3-ynyloxy)-3,4-difluoro-2-(4-iodo- 455
2-methyl-phenylamino)-benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
30 2-(4-Bromo-2-methyl-phenylamino)-N-(but- 407
3-ynyloxy)-3,4-difluoro-benzamide
31 5-Bromo-N-(but-3-ynyloxy)-3,4-difluoro- 533
2-(4-iodo-2-methyl-phenylamino)-benzamide
32 3,4-Difluoro-2-(4-iodo-2-methyl- 517
phenylamino)-N-(3-phenyl-prop-2-ynyloxy)-
benzamide
33 3,4-Difluoro-2-(4-bromo-2-methyl- 469
phenylamino)-N-(3-phenyl-prop-2-ynyloxy)-
benzamide
34 3,4-Difluoro-N-[3-(3-fluoro-phenyl)-prop- 535
2-ynyloxy]-2-(4-iodo-2-methyl-phenylamino)-
benzamide
35 2-(4-Bromo-2-methyl-phenylamino)- 487
3,4-difluoro-N-[3-(3-fluoro-phenyl)-prop-
2-ynyloxy]-benzamide
36 3,4-Difluoro-N-[3-(2-fluoro-phenyl)-prop- 535
2-ynyloxy]-2-(4-iodo-2-methyl-phenylamino)-
benzamide
37 5-Bromo-3,4-difluoro-N-[3-(2-fluoro-phenyl)- 613
prop-2-ynyloxy]-2-(4-iodo-2-methyl-
phenylamino)-benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
39 2-(4-Bromo-2-methyl-phenylamino)- 510
3,4-difluoro-N-(3-methyl-5-phenyl-pent-2-en-
4-ynyloxy)-benzamide
40 N-Ethoxy-3,4-difluoro-2-(4-iodo-2-methyl- 431
phenylamino)-benzamide
41 2-(4-Bromo-2-methyl-phenylamino)-N- 383
ethoxy-3,4-difluoro-benzamide
42 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 427
N-propoxy-benzamide
43 3,4-Difluoro-2-(4-iodo-2-methyl- 445
phenylamino)-N-propoxy-benzamide
44 2-(4-Bromo-2-methyl-phenylamino)- 397
3,4-difluoro-N-propoxy-benzamide
45 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 523
phenylamino)-N-propoxy-benzamide
46 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 427
N-isopropoxy-benzamide
47 3,4-Difluoro-2-(4-iodo-2-methyl- 445
phenylamino)-N-isopropoxy-benzamide
48 2-(4-Bromo-2-methyl-phenylamino)- 397
3,4-difluoro-N-isopropoxy-benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
49 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 523
phenylamino)-N-isopropoxy-benzamide
50 N-Cyclobutyloxy-3,4-difluoro-2-(4-iodo- 457
2-methyl-phenylamino)-benzamide
51 2-(4-Bromo-2-methyl-phenylamino)-N- 409
cyclobutyloxy-3,4-difluoro-benzamide
52 N-Cyclopentyloxy-4-fluoro-2-(4-iodo- 453
2-methyl-phenylamino)-benzamide
53 N-Cyclopentyloxy-3,4-difluoro-2-(4-iodo- 471
2-methyl-phenylamino)-benzamide
54 2-(4-Bromo-2-methyl-phenylamino)-N- 423
cyclopentyloxy-3,4-difluoro-benzamide
55 N-Cyclopropylmethoxy-4-fluoro-2-(4-iodo- 439
2-methyl-phenylamino)-benzamide
56 N-Cyclopropylmethoxy-3,4-difluoro- 457
2-(4-iodo-2-methyl-phenylamino)-benzamide
57 2-(4-Bromo-2-methyl-phenylamino)-N- 409
cyclopropylmethoxy-3,4-difluoro-benzamide
58 5-Bromo-N-cyclopropylmethoxy-3,4-difluoro- 435
2-(4-iodo-2-methyl-phenylamino)
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Example Compound Melting MS
No. Point (°C) (M-H+)
59 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 505
N-(2-phenoxy-ethoxy)-benzamide
60 3,4-Difluoro-2-(4-iodo-2-methyl- 523
phenylamino)-N-(2-phenoxy-ethoxy)-
benzamide
61 2-(4-Bromo-2-methyl-phenylamino)- 475
3,4-difluoro-N-(2-phenoxy-ethoxy)-
benzamide
62 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 481
N-(thiophen-2-ylmethoxy)-benzamide
63 3,4-Difluoro-2-(4-iodo-2-methyl- 499
phenylamino)-N-(thiophen-2-ylmethoxy)-
benzamide
64 2-(4-Bromo-2-methyl-phenylamino)- 451
3,4-difluoro-N-(thiophen-2-ylmethoxy)-
benzamide
65 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 439
N-(2-methyl-allyloxy)-benzamide
66 3,4-Difluoro-2-(4-iodo-2-methyl- 457
phenylamino)-N-(2-methyl-allyloxy)-
benzamide
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Example Compound Melting MS
No. Point (°C) (M-hi+)
67 2-(4-Bromo-2-methyl-phenylamino)- 410
3,4-difluoro-N-(2-methyl-allyloxy)-benzamide
68 N-(But-2-enyloxy)-4-fluoro-2-(4-iodo- 439
2-methyl-phenylamino)-benzamide
69 N-(But-2-enyloxy)-3,4-difluoro-2-(4-iodo- 457
2-methyl-phenylamino)-benzamide
70 2-(4-Bromo-2-methyl-phenylamino)-N-(but- 410
2-enyloxy)-3,4-difluoro-benzamide
71 3,4-Difluoro-2-(4-iodo-2-methyl- 441
phenylamino)-N-(prop-2-ynyloxy)-
benzamide
72 N-(But-3-ynyloxy)-3,4-difluoro-2-(4-iodo- 455
2-methyl-phenylamino)-benzamide
73 2-(4-Bromo-2-methyl-phenylamino)-N- 449
(4,4-dimethyl-pent-2-ynyloxy)-3,4-difluoro-
benzamide
74 N-(But-2-enyloxy)-3,4-difluoro-2-(4-iodo- 457
2-methyl-phenylamino)-benzamide
75 2-(4-Bromo-2-methyl-phenylamino)-N-(but- 410
2-enyloxy)-3,4-difluoro-benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
76 N-(3-tert.-butyl-propyn-2-yl)oxy-4-fluoro- 479
2-(4-iodo-2-methyl-phenylamino)-benzamide
77 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 577*
N-phenylmethoxy-benzamide *CI
78 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl- oil
phenylamino)-N-isopropyl-benzamide
79 N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4- 125-127
iodo-2-methyl-phenylamino)-benzamide
80 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl- 45-55
phenylamino)-N-methyl-benzamide
81 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl- 208-209
phenylamino)-5-nitro-benzamide (GLASS)
82 2-(2-Chloro-4-iodo-phenylamino)-N-hydroxy- 199-200
4-nitro-benzamide
83 3,4-Difluoro-2-(4-iodo-2-methyl- 163-165
phenylamino)-N-(tetrahydro-pyran-2-yloxy)-
benzamide
84 3,4-Difluoro-N-hydroxy-2-(4-iodo-2-methyl- 65-75
phenylamino)-benzamide
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Example Compound Melting MS
No. ~ Point (°C) (M-H+)
85 3,4-Difluoro-5-bromo-2-(4-iodo-2-methyl- 95
phenylamino)-N-(2-piperidin-1-yl-ethoxy)-
benzamide
86 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 167-169
phenylamino)-N-(tetrahydro-pyran-2-yloxy)-
benzamide
87 2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N- 165-169
hydroxy-benzamide (NCI salt)
88 2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N- 166-167.5
(tetrahydro-pyran-2-yloxy)-benzamide
89 3,4-Difluoro-2-(2-chloro-4-iodo- 173-174
phenylamino)-N-cyclobutylmethoxy-
benzamide
90 3,4-Difluoro-2-(2-chloro-4-iodo- 121-122
phenylamino)-N-(tetrahydro-pyran-2-yloxy)-
benzamide
91 5-Bromo-2-(2-chloro-4-iodo-phenylamino)- 206-211.5
N-(2-dimethylamino-ethoxy)-3,4-difluoro- DEC
benzamide monohydrochloride salt
92 5-Bromo-N-(2-dimethylamino-propoxy)-3,4- 95-105
difluoro-2-(4-iodo-2-methyl-phenylamino)-
benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
93 5-Bromo-2-(2-chloro-4-iodo-phenylamino)- 266-280
3,4-difluoro-N-hydroxy-benzamide DEC
94 5-Bromo-2-(2-chloro-4-iodo-phenylamino)- 167.5-169.5
3,4-difluoro-N-(tetrahydro-pyran-2-yloxy)-
benzamide
95 3,4-Difluoro-2-(2-chloro-4-iodo- 172.5-173.5
phenylamino)-N-cyclopropylmethoxy-
benzamide
96 5-Bromo-2-(2-chloro-4-iodo-phenylamino)- 171-172.5
N-cyclopropylmethoxy-3,4-difluoro-
benzamide
97 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 173.5-175
phenylamino)-N-(2-morpholin-4-yl-ethoxy)-
benzamide
98 5-Bromo-N-(2-diethylamino-ethoxy)-3,4- 81 DEC
difluoro-(4-iodo-2-methyl-phenylamino)-
benzamide
99 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 126-128
phenylamino)-N-isobutoxy-benzamide
100 5-Bromo-N-cyclohexylmethoxy-3,4-difluoro- 139-142
2-(4-iodo-2-methyl-phenylamino)-benzamide
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Example Compound Melting MS
No. Point (°C) (M-H+)
101 5-Bromo-N-cyclopentylmethoxy-3,4-difluoro- 113-115
2-(4-iodo-2-methyl-phenylamino)-benzamide
102 5-Bromo-N-cyclobutylmethoxy-3,4-difluoro- 138-139
2-(4-iodo-2-methyl-phenylamino)-benzamide
The invention compounds are useful in treating chronic pain
proliferative diseases by virtue of their selective inhibition of the dual
specificity protein kinases MEK1 and MEK2. The invention compound has
been evaluated in a number of biological assays which are normally utilized to
establish inhibition of proteins and kinases, and to measure mitogenic and
metabolic responses to such inhibition.
EXAMPLE 1A
4-Fluoro-2-(4-iodo-2-methylphenylamino)benzoic acid
To a stirring solution comprised of 3.16 g (0.0133 mol) of 2-amino-
5-iodotoluene in 5 mL of tetrahydrofuran at -78°C was added 10 mL
(0.020 mol) of a 2.0 M lithium diisopropylamide in
tetrahydrofuran/heptane/ethenylbenzene (Aldrich) solution. The resulting
green suspension was stirred vigorously for 15 minutes, after which time a
solution of 1.00 g (0.00632 mol) of 2,4-difluorobenzoic acid in 10 mL of
tetrahydrofuran was added. The reaction temperature was allowed to increase
slowly to room temperature, at which temperature it was stirred for 2 days.
The reaction mixture was concentrated. Aqueous HCI (10%) was added to the
concentrate, and the solution was extracted with dichloromethane. The
organic phase was dried (MgS04) and then boiled over a steambath to low
volume and cooled to room temperature. The off-white fibers were collected
by vacuum filtration, rinsed with hexanes, and vacuum-oven dried.
(76°C; ca.
10 mm of Hg) to afford 1.10 g (47%) of the desired material; mp 224-
229.5°C;
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1 H NMR (400 MHz; DMSO): 8 9.72 (s, 1 H), 7.97 (dd, 1 H, J = 7.0, 8.7 Hz),
7.70 (d, 1 H, J = 1.5 Hz), 7.57 (dd, 1 H, J = 8.4, 1.9 Hz), 7.17 (d, 1 H,
J = 8.2 Hz), 6.61-6.53 (m, 2H), 2.18 (s, 3H);
13C NMR (100 MHz; DMSO): 8 169.87, 167.60, 165.12, 150.17, 150.05,
139.83, 138.49, 136.07, 135.31, 135.20, 135.07, 125.60, 109.32, 105.09,
104.87, 99.72, 99.46, 89.43, 17.52;
19F NMR (376 MHz; DMSO): 8 -104.00 to -104.07 (m);
IR (KBr) 1670 (C = O stretch) cm-1;
MS (CI) M+1 = 372.
Analysis calculated for C14H11 FIN02:
C, 45.31; H, 2.99; N, 3.77.
Found: C, 45.21; H, 2.77; N, 3.64.
EXAMPLES 2A-30A
By following the general procedure of Example 1A, the following
benzoic acids and salts were prepared:
Example Compound MP °C
No.
2A 3,4,5-Trifluoro-2-(4-iodo-2-methyl-phenylamino)- 206-210
benzoic acid
3A 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 240.5-244.5
benzoic acid
4A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 259.5-262
phenylamino)-benzoic acid
5A 5-Chloro-2-(2-chloro-4-iodo-phenylamino)-benzoic 255-260
acid
6A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic 234-238
acid
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Example Compound MP °C
No.
7A Sodium 5-Chloro-2-(4-iodo-2-methyl-phenylamino)- 310-320 DE
benzoate C
8A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-benzoic 239.5-240
acid
9A 2-(2-Chloro-4-iodo-phenylamino)-5-nitro-benzoic 289-293
acid
10A 4-Fluoro-2-(3-fluoro-4-iodo-2-methyl-phenylamino)- 233-235
benzoic acid
11A 2-(4-lodo-2-methyl-phenylamino)-5-vitro-benzoic 264-267
acid
12A 2-(2-Fluoro-4-iodo-phenylamino)-5-vitro-benzoic 256-258
acid
13A 2-(4-Bromo-2-methyl-phenylamino)-4-fluoro- 218.5-220
benzoic acid
14A 2-(2-Bromo-4-iodo-phenylamino)-5-vitro-benzoic 285-288 DE
acid C
15A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro- 230-234
benzoic acid
16A 3-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic 218-221
acid
17A 3,4-Difluoro-2-(4-iodo-2-methoxy-phenylamino)- 230-233
benzoic acid
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Example Compound MP °C
No.
18A 4-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic 245-255 DE
acid C
19A 2-(4-lodo-2-methyl-phenylamino)-benzoic acid 218-223
20A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic 243-46
acid
21A 5-lodo-2-(4-iodo-2-methyl-phenylamino)-benzoic 241-245
acid
26A 2-Fluoro-6-(4-iodo-2-methyl-phenylamino)-benzoic 179-182
acid
27A 4-Fluoro2-(2,3-dimethyl-4-iodo-2-methyl- 258-261
phenylamino)-benzoic acid
28A 5-Methyl-2-(4-iodo-2-methyl-phenylamino)-benzoic 209.5-211
acid
29A 2-Chloro-6-(4-iodo-2-methyl-phenylamino)-benzoic 171-175
acid
30A 2-(4-lodo-2-methyl-phenylamino)-4-nitro-benzoic 251-263
acid
EXAMPLE 31A
5-Chloro-N-(2-h dy roxyethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide
To a stirring solution comprised of 0.1020 g (0.2632 mmol) of 5-chloro-
2-(4-iodo-2-methyl-phenylamino)-benzoic acid, 0.1 mL (1.7 mmol) of
ethanolamine, and 0.05 mL (0.29 mmol) of diisopropylethylamine in 5 mL of a
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1:1 (v/v) tetrahydrofuran-dichloromethane solution was added 0.15 g
(0.29 mmol) of solid PyBOP powder directly. The reaction mixture was stirred
at room temperature overnight. The solvent was removed in vacuo. The crude
residue was partitioned between ether (50 mL) and 10% aqueous hydrochloric
acid (50 mL). The organic phase was washed with 10% aqueous sodium
hydroxide (50 mL), dried (MgS04) and concentrated in vacuo to afford a
yellow-brown oil which was crystallized from hexanes-ether to afford 0.0831 g
(73%) of a green-yellow powder; mp 120-121°C;
1 H NMR (400 MHz; CDC13): 8 9.11 (s, 1 H); 7.56 (d, 1 H, J = 1.4 Hz),
7.46-7.41 (m, 2H), 7.20 (dd, 1 H, J = 8.9, 2.4 Hz), 7.00 (t, 2H, J = 9.6 Hz),
6.55 (broad t, 1H), 3.86 (t, 2H, J = 5.0 Hz), 3.61 (dd, 2H, J = 10.1, 5.5 Hz),
2.23 (s, 3H), 1.56 (broad s, 1 H);
IR (KBr) 3297 (O-H stretch), 1627 (C = O stretch) cm-1;
MS (CI) M+1 = 431.
Analysis calculated for ClgHIgCIIN202:
C, 44.62; H, 3.74; N, 6.50.
Found: C, 44.63; H, 3.67; N, 6.30.
EXAMPLES 32-48A
By following the general procedure of Example 31A, the following
benzamides were prepared by reacting the corresponding benzoic acid with
the corresponding amine.
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Example Compound MP C
No.
32A 4-Methoxy-N-(4-methoxy-phenyl)-3-nitro-153.5-156
benzamide
33A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-158
benzamide
34A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-102.5-104.5
methyl-benzamide
35A N-Ethyl-4-fluoro-2-(4-iodo-2-methyl-90-91
phenylamino)-benzamide
36A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N,N-oil
dimethyl-benzamide
37A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-285-288
DE
(1 H-tetrazol-5-yl)-benzamide C
38A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-180-182
benzamide
39A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N,N-137-138
dimethyl-benzamide
40A [5-Chloro-2-(4-iodo-2-methyl-phenylamino)-170-173
benzoylamino]-acetic acid
41A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-69-71
propyl-benzamide
42A 5-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-132-133.4
2-methyl-phenylamino)-benzamide
43A N,N-Diethyl-4-fluoro-2-(4-iodo-2-methyl-oil
phenylamino)-benzamide
44A 4-Fluoro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-122-124
1-yl]-propyl}-2-(4-iodo-2-methyl-phenylamino)-
benzamide
45A N,N-Diethyl-2-(4-iodo-2-methyl-phenylamino)-91-93
5-nitro-benzamide
46A N-Butyl-4-fluoro-2-(4-iodo-2-methyl-97-99
phenylamino)-benzamide
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Example Compound MP °C
No.
47A 5-Chloro-N,N-diethyl-2-(4-iodo-2-methyl- 118-120
phenylamino)-benzamide
48A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N,N- 142.5-144
dimethyl-benzamide
EXAMPLE 49A
A4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzyl alcohol4-Fluoro-
2-(4-iodo-2-methyl-phenylamino)-benzoic acid (0.50 g, 1.35 mmol) was
dissolved in 6 mL (6 mmol) of cold 1.0 M borane-tetrahydrofuran complex in
tetrahydrofuran solution. The reaction mixture was stirred under nitrogen
atmosphere at room temperature overnight. The reaction was quenched with
80 mL of methanol. Concentration in vacuo produced a clear tan oil which was
purified by MPLC. Elution with dichloromethane afforded 0.4285 g (89%) of a
white solid; mp 99-100.5°C;
1 H NMR (400 MHz; DMSO): 8 7.57 (d, 1 H, J=1.7 Hz), 7.45 (dd, 1 H, J=8.4,
1.9 Hz), 7.39 (s, 1 H), 7.29 (t, 1 H, J=7.5 Hz), 6.89 (d, 1 H, J=8.4 Hz),
6.67-6.60 (m, 1 H), 5.47 (t, 1 H, J=5.5 Hz), 4.49 (d, 2H, 5.1 Hz), 2.14 (s,
3H);
IR (KBr) 3372 (O-H stretch) cm-1;
MS (CI) M+1 = 358.
Analysis calculated for C14H13FIN0:
C, 47.08; H, 3.67; N, 3.92.
Found: C, 47.17; H, 3.75; N, 3.72.
EXAMPLE 50A-52A
The following benzyl alcohols were prepared by the general procedure
of Example 49A.
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Example Compound MP °C
No.
50A [5-Chloro-2-(4-iodo-2-methyl-phenylamino)- 82-85
phenyl]-methanol
51A [2-(4-lodo-2-methyl-phenylamino)-5-nitro- 126.5-128.
phenyl]-methanol 5
52A [5-Bromo-2-(4-iodo-2-methyl-phenylamino)- 60.5-63.5
phenyl]-methanol
Several invention compounds of Formula I(A) were prepared utilizing
combinatorial synthetic techniques. The general procedure is as follows:
To a 0.8-mL autosampler vial in a metal block was added 40 ~L of a
0.5 M solution of the acid in DMF and 40 p,L of the reagent amine (2M solution
in Hunig's base and 1 M in amine in DMF). A 0.5M solution of PyBrop was
freshly prepared and 50 ~L were added to the autosampler vial. The reaction
was allowed to stand for 24 hours.
The reaction mixture was transferred to a 2-dram vial and diluted with
2 mL of ethyl acetate. The organic layer was washed with 3 mL of distilled
water and the water layer washed again with 2 mL of ethyl acetate. The
combined organic layers were allowed to evaporate to dryness in an open
fume hood.
The residue was taken up in 2 mL of 50% acetonitrile in water and
injected on a semi-prep reversed phase column (10 mm x 25 cm, 5 pM
spherical silica, pore size 115 A derivatized with C-18, the sample was eluted
at 4.7 mL/min with a linear ramp to 100% acetonitrile over 8.5 minutes.
Elution
with 100% acetonitrile continued for 8 minutes). Fractions were collected by
monitoring at 214 nM. The residue was dissolved in chloroform and
transferred to a preweighed vial, evaporated, and weighed again to determine
the yield.
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EXAMPLES 53A-206A
The following compounds of Formula I were prepared by combinatorial
methodology:
Example Compound MS
No. M-H
53A 5-Bromo-3,4-difluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-510
2-methyl-phenylamino)-benzamide
54A N-(2,3-Dihydroxy-propyl)-3,4-difluoro-2-(4-iodo-2-methyl-462
phenylamino)-benzamide
55A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-577
(2-piperidin-1-yl-ethyl)-benzamide
56A 3,4-Difluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-432
phenylamino)-benzamide
57A N-(2,3-Dihydroxy-propyl)-4-fluoro-2-(4-iodo-2-methyl-444
phenylamino)-benzamide
58A 3,4-Difluoro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-446
phenylamino)-benzamide
59A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-564
(2-pyrrolidin-1-yl-ethyl)-benzamide
60A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-571
(2-pyridin-4-yl-ethyl)-benzamide
61A 4-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-414
phenylamino)-benzamide
62A 5-Bromo-N-(3-dimethylamino-propyl)-3,4-difluoro-2-(4-iodo-551
2-methyl-phenylamino)-benzamide
63A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-580
(2-morpholin-4-yl-ethyl)-benzamide
64A 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-501
(2-morpholin-4-yl-ethyl)-benzamide
65A 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-485
(2-pyrrolidin-1-yl-ethyl)-benzamide
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Example Compound MS
No. M-H
66A 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-493
4-yl-ethyl)-benzamide
67A N-(3-Dimethylamino-propyl)-3,4-difluoro-2-(4-iodo-2-methyl-473
phenylamino)-benzamide
68A N-Benzyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-460
benzamide
69A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-384
(2-hydroxy-ethyl)-benzamide
70A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-483
4-yl-ethyl)-benzamide
71A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-495
1-yl-propyl)-benzamide
72A 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-513
1-yl-propyl)-benzamide
73A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-thiophen-480
2-yl-ethyl)-benzamide
74A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-467
1-yl-ethyl)-benzamide
75A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-453
(2-morpholin-4-yl-ethyl)-benzamide
76A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-557
pyridin-4-ylmethyl-benzamide
77A 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-479
4-ylmethyl-benzamide
78A 2-(4-Bromo-2-methyl-phenylamino)-N-(3-dimethylamino-425
propyl)-3,4-difluoro-benzamide
79A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-461
4-ylmethyl-benzamide
80A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-4-yl-475
ethyl)-benzamide
81A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-pyridin-445
4-yl-ethyl)-benzamide
82A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-400
(3-hydroxy-propyl)-benzamide
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Example Compound MS
No. M-H
83A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-437
(2-pyrrolidin-1-yl-ethyl)-benzamide
84A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-phenethyl-474
benzamide
85A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-450
(2-thiophen-2-yl-ethyl)-benzamide
86A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-pyridin-431
4-ylmethyl-benzamide
87A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-444
phenethyl-benzamide
88A 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-451
(2-piperidin-1-yl-ethyl)-benzamide
89A 5-Chloro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-557*
2-(4-iodo-2-methyl-phenylamino)-benzamide
90A 5-Fluoro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-541*
2-(4-iodo-2-methyl-phenylamino)-benzamide
91A 2-(4-lodo-2-methyl-phenylamino)-5-nitro-N-pyridin-4-yl487
methyl-benzamide
92A 5-Bromo-N-(3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-601*
2-(4-iodo-2-methyl- phenylamino)-benzamide
93A 5-Chloro-N-(2-diethylamino-ethyl)-2-(4-iodo-2-methyl-486*
phenylamino)-benzamide
94A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-497*
1-yl-ethyl)-benzamide
95A (3-Hydroxy-pyrrolidin-1-yl)-[2-(4-iodo-2-methyl-466
phenylamino)-5-nitro-phenyl]-
96A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-484*
1-yl-ethyl)-benzamide
97A 5-Bromo-N-(2-diethylamino-ethyl)-2-(4-iodo-2-methyl-530*
phenylamino)-benzamide
98A N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-chloro-2-(4-iodo-518*
2-methyl- phenylamino)-benzamide
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Example Compound MS
No. M-H
99A N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-bromo-2-(4-iodo-562*
2-methyl- phenylamino)-benzamide
100A [5-Bromo-2-(4-iodo-2-methyl-phenylamino)-phenyl]-499
(3-hydroxy-pyrrolidin-1-yl)-
101A 2-(4-lodo-2-methyl-phenylamino)-5-nitro-benzoic501
acid
phenethyl ester
102A N-{3-[4-(2-Hydroxy-ethyl)-piperazin-1-yl]-propyl}-2-(4-iodo-568*
2-methyl-phenylamino)-benzamide
103A [5-Chloro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-455
(3-hydroxy-pyrrol idi n-1-yl)-
104A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-460
4-ylmethyl-benzamide
105A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-528*
1-yl-ethyl)-benzamide
106A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-542*
1-yl-ethyl)-benzamide
107A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-468*
1-yl-ethyl)-benzamide
108A 5-Chloro-N-(3-dimethylamino-propyl)-2-(4-iodo-2-methyl-472*
phenylamino)-benzamide
109A N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-fluoro-2-(4-iodo-502*
2-methyl-phenylamino)-benzamide
110A 5-Chloro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-445*
phenylamino)-benzamide
111A 5-Chloro-N-(3-diethylamino-2-hydroxy-propyl)-2-(4-iodo-516*
2-methyl-phenylamino)-benzamide
112A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-482*
1-yl-ethyl)-benzamide
113A 5-Bromo-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-489*
phenylamino)-benzamide
114A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-556*
1-yl-propyl)-benzamide
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Example Compound MS
No. M-H
115A N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-2-(4-iodo-2-methyl-529*
phenylamino)-5-vitro-benzamide
116A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-500*
4-yl-ethyl)-benzamide
117A 5-Chloro-N-(3-diethylamino-propyl)-2-(4-iodo-2-methyl-500*
phenylamino)-benzamide
118A 5-Chloro-N-(2-diisopropylamino-ethyl)-2-(4-iodo-2-methyl-514*
phenylamino)-benzamide
119A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-512*
1-yl-propyl)-benzamide
120A 2-(4-lodo-2-methyl-phenylamino)-5-vitro-N-(2-piperidin-1-yl-509*
ethyl)-benzamide
121A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperazin-544*
1-yl-ethyl)-benzamide
122A N-(2-Diethylamino-ethyl)-5-fluoro-2-(4-iodo-2-methyl-470*
phenylamino)-benzamide
123A 5-Bromo-N-(3-dimethylamino-propyl)-2-(4-iodo-2-methyl-516*
phenylamino)-benzamide
124A N-(3-Hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-456*
5-vitro-benzamide
125A 5-Fluoro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-429*
phenylamino)-benzamide
126A N-(3-Diethylamino-propyl)-5-fluoro-2-(4-iodo-2-methyl-484*
phenylamino)-benzamide
127A N-(3-Diethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)-511*
5-vitro-benzamide
128A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-544*
4-yl-ethyl)-benzamide
129A 2-(4-lodo-2-methyl-phenylamino)-5-vitro-N-(3-piperidin-1-yl-523*
propyl)-benzamide
130A [5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-439
(3-hydroxy-pyrrolidin-1-yl)-
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Example Compound MS
No. M-H
131A 5-Bromo-N-(2-diisopropylamino-ethyl)-2-(4-iodo-2-methyl-558*
phenylamino)-benzamide
132A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-484*
4-yl-ethyl)-benzamide
133A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-496*
1-yl-propyl)-benzamide
134A [5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-482
[4-(2-hyd roxy-ethyl)-piperazin-1-
135A N-(3-Diethylamino-2-hydroxy-propyl)-5-fluoro-2-(4-iodo-500*
2-methyl-phenylamino)-benzamide
136A [5-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoylamino]-443
acetic acid
137A 2-(4-lodo-2-methyl-phenylamino)-5-vitro-N-(2-pyrrolidin-1-yl- 495*
ethyl)-benzamide
138A N-(3-Dimethylamino-propyl)-2-(4-iodo-2-methyl-483*
phenylamino)-5-vitro-benzamide
139A N-(2-Diisopropylamino-ethyl)-5-fluoro-2-(4-iodo-2-methyl-498*
phenylamino)-benzamide
140A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic490
acid
S-phenethyl ester
141A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic506
acid
S-phenethyl ester
142A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic536
acid
S-benzyl ester
143A 2-(4-lodo-2-methyl-phenylamino)-5-vitro-thiobenzoic503
acid
S-benzyl ester
144A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic476
acid
S-benzyl ester
145A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic492
acid
S-benzyl ester
146A N-Cyclopropyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-409
benzamide
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Example Compound MS
No. M-H
147A 5-Chloro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-429
~
phenylamino)-benzamide
148A 5-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-413
phenylamino)-benzamide
149A N-Benzyloxy-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-475
benzamide
150A N-Benzyloxy-5-bromo-2-(4-iodo-2-methyl-phenylamino)-593*
benzamide
151A 2-(4-lodo-2-methyl-phenylamino)-5-vitro-N-(4-sulfamoyl-567
benzyl)-benzamide
152A 5-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-473
phenylamino)-benzamide
153A N-(2-Hydroxy-ethyl)-5-iodo-2-(4-iodo-2-methyl-521
phenylamino)-benzamide
154A N-(2-Hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-440
5-vitro-benzamide
155A 2-(4-lodo-2-methyl-phenylamino)-N-methyl-5-vitro-N-phenyl-486
benzamide
156A 5-Chloro-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-425
benzamide
157A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-459
phenyl-benzamide
158A N-Allyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide409
159A N-Benzyloxy-5-iodo-2-(4-iodo-2-methyl-phenylamino)-583
benzamide
160A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-538
benzyl)-benzamide
161A N-Allyl-5-chloro-2-(4-iodo-2-methyl-phenylamino)-425
benzamide
162A N-Cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-5-vitro-436
benzamide
163A 5-Bromo-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-469
benzamide
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Example Compound MS
No. M-H
164A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-475
phenyl-benzamide
165A 5-lodo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-646
benzyl)-benzamide
166A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-598
benzyl)-benzamide
167A N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide436
168A 2-(4-lodo-2-methyl-phenylamino)-5-nitro-N-(4-sulfamoyl-565
benzyl)-benzamide
169A N-Allyl-5-bromo-2-(4-iodo-2-methyl-phenylamino)-469
benzamide
170A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-473
benzyl)-benzamide
171A N-Cyclopropyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-517
benzamide
172A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-519
phenyl-benzamide
173A N-Benzyloxy-2-(4-iodo-2-methyl-phenylamino)-5-nitro-502
benzamide
174A N-Cyclohexyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-559
benzamide
175A N-Allyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide517
176A 5-lodo-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-581
benzyl)-benzamide
177A 2-(4-lodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-500
5-vitro-benzamide
178A 5-lodo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-567
benzamide
179A N-Cyclohexyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-451
benzamide
180A 5-Chloro-N-cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-467
benzamide
181A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-533
benzyl)-benzamide
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Example Compound MS
No. M-H
182A 5-Bromo-N-cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-511
benzamide
183A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-489
benzyl)-benzamide
184A N-Cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-5-vitro-478
benzamide
185A N-Benzyloxy-5-bromo-2-(4-iodo-2-methyl-phenylamino)-538
benzamide
186A N-Benzyloxy-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-477
benzamide
187A 5-Chloro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-431
phenylamino)-benzamide
188A 5-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-475
phenylamino)-benzamide
189A 2-(4-lodo-2-methyl-phenylamino)-N-methyl-5-vitro-N-phenyl-488
benzamide
190A 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-477
phenyl-benzamide
191A N-(2-Hydroxy-ethyl)-5-iodo-2-(4-iodo-2-methyl-523
phenylamino)-benzamide
192A 5-Chloro-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-425
benzamide
193A N-Allyl-5-chloro-2-(4-iodo-2-methyl-phenylamino)-427
benzamide
194A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-461
phenyl-benzamide
195A N-(2-Hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-442
5-vitro-benzamide
196A 5-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-415
phenylamino)-benzamide
197A 5-Bromo-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-472
benzamide
198A N-Cyclopropyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-411
benzamide
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Example Compound MS
No. M-H
199A 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-540
benzyl)-benzamide
200A N-Cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-438
benzamide
201A N-Allyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide411
202A N-Benzyloxy-5-iodo-2-(4-iodo-2-methyl-phenylamino)-585
benzamide
203A N-Allyl-5-bromo-2-(4-iodo-2-methyl-phenylamino)-472
benzamide
204A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-601
benzyl)-benzamide
205A 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-522
phenyl-benzamide
206A N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide438
* M+H
EXAMPLE 207A
Preparation of f4-Chloro-2-(1 H-tetrazol-5-yl)-(4-iodo-2-methyl-phenyl)-amine
Step a: Preparation of 5-Chloro-2-fluoro-benzaldehyde
To a solution of 1-chloro-4-fluorobenzne (13.06 g, 0.1 mol) in THF
(180 mL), at -78°C, LDA (2 M solution in THF, 50 mL, 0.1 mol) was added
dropwise. After stirring at -78°C for 1.5 hours, DMF (8 mL) was added
to the
reaction mixture and allowed to warm up to room temperature overnight. The
reaction mixture was partitioned between water and Et20. The Et20 layer
was dried (MgS04) and the solvent removed in vacuum to give 14.95 g (94%)
yield of crude aldehyde:
1 H NMR (CDC13): 8, 10.3 (s, -C(=O)H).
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Step b: Preparation of 5-Chloro-2-fluoro-benzaldehyde oxime
A solution of 5-chloro-2-fluoro-benzaldehyde (10 g, 0.0631 mol),
hydroxylamine hydrochloride (6.57 g, 0.0946 mol) and pyridine (8.3 mL,
0.1010 mol) in EtOH (100 mL) was heated at 75°C (oil bath temperature)
for
1 hour and the solvent removed under vacuum to give an oil. The oil was
partitioned between water and CH2C12. The CH2C12 layer was dried (MgS04)
and the solvent removed under vacuum to give crude aldoxime as a solid. The
solid was purified by medium pressure liquid chromatography on silica. Elution
with CH2C12 gave 4.87 g (28%) of the aldoxime as white solid: mp 95-
97°C;
Analysis calculated for C7H5NOFC1:
C, 48.44; H, 2.90; N, 8.07.
Found: C, 48.55; H, 2.69, N, 7.90.
Step c: Pre~~aration of 5-Chloro-2-fluoro-benzonirile
A solution of the 5-chloro-2-fluoro-benzaldehyde oxime (3.15 g,
0.0182 mol) in acetic anhydride (150 mL) was refluxed for 16 hours. The
reaction mixture was cooled to room temperature and poured into saturated
aqueous NaHC03 (200 mL) solution. The mixture was extracted with Et20.
The Et20 layer was dried (K2C03) and the solvent removed to give the
product as an oily solid. The product was used without further purification in
the next step.
Step d: Preparation of 5-(5-Chloro-2-fluoro-phenyl)-1 H-tetrazole
A mixture of 5-chloro-2-fluoro-benzonitrile (2.84 g, 0.01823 mol),
butanol (15 mL), sodium azide (1.543 g, 0.0237 mol), acetic acid (1.36 mL,
0.0237 mol) was refluxed for 24 hours. The reaction mixture was cooled to
room temperature, additional 1.543 g sodium azide added, and the reaction
mixture refluxed for additional 24 hours. After cooling to room temperature,
Et20 (100 mL) and 10% aqueous NaOH (200 mL) were added sequentially.
The mixture was vigorously stirred. The aqueous layer was separated, cooled
with ice-methanol bath (-15°C) and acidified to pH 1 with conc. NCI. A
gray
solid precipitated. The solid was dried in vacuum at 50°C to give 1.76
g (49%)
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of 5-(5-chloro-2-fluoro-phenyl)-1 H-tetrazole: mp partial melt at
110°C,
complete melting at 124°C);
1 H (400 Mz, CDC13): 8 8.19-8.08 (m, 1 H), 7.77-7.71 (m, 1 H), 7.61-7.52 (m,
1 H); 13C (100 Mz, CDC13): 8 159.00, 156.49, 140.88, 133.02, 132.93, 130.73,
129.23, 129.21, 129.08, 126.05, 118.96, 118.73, 114.50;
MS (CI) M+1 = 199 (100), M = 198 (6).
Step e: Preparation of f4-Chloro-2-1~1 H-tetrazol-5-yl)-(4-iodo-2-methyl-
phenyl)-
amine
To a solution of 2-methyl-4-iodoaniline (3.52 g, 0.0151 mol) in THF
(25 mL) at -78°C, LDA (2 molar solution in THF, 11.33 mL, 0.02267 mol)
was
added dropwise. After stirring for 0.5 hours, a solution of 1-(tetrazol-5-yl)-
2-fluoro-5-chlorobenzene (1.5 g, 0.00756 mol) in THF (15 mL) was added
dropwise. The reaction was stirred for 16 hours as it warmed up to room
temperature. The reaction mixture was quenched with aqueous conc. NH4C1
solution and extracted with CH2C12. The organic layer was dried (MgS04)
and the solvent removed giving a crude product as an oil. The oil with
CH2C12->CH2C12:MeOH (9.7:0.3) gave 1.5 g (48%) of the desired product:
mp 205-208; 1 H (400 Mz, DMSO): 8 9.13 (s, 1 H), 8.00-7.99 (s, 1 H), 7.69 (s,
1 H), 7.55-7.52 (m, 1 H), 7.43-7.40 (m, 1 H), 7.12-7.05 (m, 1 H), 2.24 (s,
3H);
13C (100 Mz, CDC13): 8 141.87, 139.28, 138.88, 135.47, 133.71, 131.65,
128.15, 123.69, 121.94, 116.68, 87.79, 17.22;
MS (CI) M+2 = 413 (44), M+1 = 412 (85), M = 411 (100).
Analysis calculated for C14H11N5C11~0.5H20:
C, 39.97; H, 2.87; N, 16.65.
Found: C, 38.87, H, 2.77; N, 16.47.
The following tetrazole substituted phenylamines were prepared by
following the general procedure of Example 207A.
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EXAMPLE 208A
(4-lodo-2-methyl-~~henyl)-[2-(1 H-tetrazol-5-yl)-phenyllamine, mp 231
°C (dec)
EXAMPLE 209A
[4-Nitro-2-(1 H-tetrazol-5-yl)-(4-iodo-2-methyl-phenyl)-amine, mp 205-
208°C.
EXAMPLES 210A-224A
Additional invention compounds which were prepared by the general methods
described above are:
Example Compound MP C
No.
210A 2-(2-Chloro-4-iodo-phenylamino)-3-fluoro-4-(2-239-241
morpholin-4-yl-ethylamino)-5-nitro-benzoicDEC
acid
211A 4-Amino-2-(2-chloro-4-iodo-phenylamino)-3->270
fluoro-5-nitro-benzoic acid
212A 2,4-Bis-(2-chloro-4-iodo-phenylamino)-3-fluoro->265 DEC
5-nitro-benzoic acid
213A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-5-218-225
vitro-benzoic acid DEC
214A 2-(2,6-Difluoro-4-iodo-phenylamino)-3,4-247-249
difluoro-benzoic acid
215A 2-(2-Chloro-4-iodo-phenylamino)-4-vitro-267-269
benzoic acid
216A 2-(2,4-Diiodo-phenylamino)-4-fluoro-benzoic260-261
acid
217A 2-(2-Bromo-4-iodo-phenylamino)-4-fluoro-259-262
benzoic acid
218A 4-Fluoro-2-(2-fluoro-4-iodo-phenylamino)-215-217
benzoic acid
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Example Compound MP °C
No.
219A 2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-242-247
benzoic acid
220A 5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-312.5-318
difluoro-benzoic acid
221A 2,3,5-Trifluoro-6-(4-iodo-2-methyl-phenylamino)-118-121
4-(4-methyl-piperazin-1-yl)-benzoic
acid methyl
ester dihydrofluoride salt
222A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-214-217
phenylamino)-N-(4-methyl-piperazin-1-yl)-DEC
benazmide
223A 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-154-175
phenylamino)-benzoic acid N',N'-dimethyl-DEC
hydrazide
224A 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-153.5-156
benzoic acid hydrazide
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F. Other Embodiments
From the above disclosure and examples, and from the claims below,
the essential features of the invention are readily apparent. The scope of the
invention also encompasses various modifications and adaptations within the
knowledge of a person of ordinary skill. Examples include a disclosed
compound modified by addition or removal of a protecting group, or an ester,
pharmaceutical salt, hydrate, acid, or amide of a disclosed compound.
Publications cited herein are hereby incorporated by reference in their
entirety.
What is claimed is:
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