Note: Descriptions are shown in the official language in which they were submitted.
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NITROGEN HETEROCYCLE BIARYLS
FOR OSTEOPOROSIS AND OTHER DISEASES
Field of the Invention
[0001] The invention relates to a genus of nitrogen heterocycle biaryls having
a
carboxylate terminus. The nitrogen heterocycle biaryls are useful for treating
endometri.osis, osteoporosis, restenosis following angioplasty, rheumatoid
arthritis,
cancer, macular degeneration and obesity.
Background of the Invention
[0002] Integrins are a superfamily of heterodimeric transmembrane
glycoproteins that function in cellular adhesion, migration and signal
transduction.
These glycoproteins consist of an a and a(3-subunit, which associate non-
covalently
in defined combinations. To date, 17 a-subunits and eight 0-subunits have been
identified, which associate selectively to form at least 23 integrins, all of
which
appear to function as receptors.
[0003] The integrin aV(33, also referred to as the vitronectin receptor, is
expressed on a variety of cell types, including osteoclasts, vascular smooth
muscle
cells, endothelial cells and various tumor cells. In general, the level of
expression
of av(33 integrin is low on most cell types but it is greatly increased in
remodeling or
growing tissues. Consistent with its expression profile, aV(33 integrin
mediates
several biologically relevant processes, such as adhesion of osteoclasts to
bone,
vascular smooth muscle cell migration and angiogenesis. As a result,
vitronectin
receptor antagonists are useful for the treatment of endometriosis,
osteoporosis,
restenosis following percutaneous transluminal coronary angioplasty (PTCA),
rheumatoid arthritis, cancer and ocular diseases [see, e.g., Miller et al.,
Identification and in vivo efficacy of srnall-naolecule antagonists of
integrin a,,Q3
(the vitronectin receptoY), Drug Discovery Today, Vol, 5, No. 9, 397-408
(2000)].
= 10004] Osteoporosis is a debilitating bone disease characterized by a
decrease in
bone mass (osteopenia) leading to an increased risk of fracture. The
osteopenia
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associated with osteoporosis arises from an imbalance between bone resorption
and
formation, such that resorption exceeds formation. For bone resorption to
occur,
the bone-resorbing osteoclasts must first adhere to the bone matrix and this
key
adhesive event is mediated by av(33 integrin. Disruption of osteoclast
adhesion
inhi.bits bone resorption both in vitro and in vivo and provides a therapeutic
approach to the treatment and/or prevention of osteoporosis [see, Miller et
al.
(2000)].
[0005] Angiogenesis (the fornlation of new blood vessels) involves the av[33
integrin mediated migration and proliferation of endothelial cells.
Significantly, the
vitronectin receptor is upregulated only in growing vessels and not in mature
ones.
Thus, vitronectin antagonists are useful in the treatment of diseases
characterized by
excessive or undesirable angiogenesis, such as cancer, rheumatoid arthritis,
diabetic
retinopathy and macular degeneration. In support of this hypothesis,
vitronectin-
selective antibodies and peptides have been shown to be effective inhibitors
of
angiogenesis [see, Miller et al. (2000)].
100061 US patent 6,306,819 describes the use of angiogenesis inhibitors in
vivo
for obesity, intestinal polyps, cardiac hypertrophy, and endometriosis.
Initial studies
conducted in genetically obese mice showed that inhibition of angiogenesis led
to
reduction in adipose tissue mass. Weight gain in animals receiving
angiogenesis
inhibitors was significantly reduced, in spite of increases in appetite
sufficient to
cause weight gain in paired-fed mice. Discontinuation of the inhibitor
resulted in
rapid expansion of the adipose tissue. The effect was dose-dependent,
repeatedly
reversible, and occurred in response to all of the inhibitors tested.
Significant
inhibition was also observed in both the intestinal polyp and cardiac
hypertrophy
animal models. Results in vivo in an endometriosis model also showed decreased
development of endometriosis in animals treated with angiogenesis inhibitors.
[0007] Rheumatoid arthritis is a debilitating, systemic autoimmune disease in
which there is massive bone and cartilage destruction within articulating
joints.
Integrin av[i3 is expressed on the vessels within the invasive pannus and
appears to
play a role in angiogenic vessel formation within the highly invasive
hypertrophic
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synovium. In addition, vitronectin mediates the bone resorption process.
Because
rheumatoid arthritis involves both angiogenic vessel formation and bone
resorption,
vitronectin antagonists provide therapy for rheumatoid arthritis. A cyclic
avP3
integrin inhibitor (an Arg-Gly-Asp peptide) has been shown to be effective in
vivo
in a rabbit model of inflammatory arthritis [see, Miller et al. (2000)].
[0008] Restenosis refers to a significant, delayed loss of blood vessel lumen
that
generally occurs after percutaneous transluminal coronary angioplasty (PTCA).
Vascular smooth muscle cell migration into the neointima is a necessary step
in
restenosis and av(33 integrin, which is expressed on smooth muscle cells, has
been
shown to mediate this migration. In addition, vascular injury induced by PTCA
causes a rapid, persistent and coordinated upregulation of av(Is , avPs and
osteopontin during the period of neointimal development. Studies have shown
that
blocking the vitronectin receptor inhibits smooth muscle cell migration and
that
both peptide antagonists and a humanized monoclonal antibody are effective in
reducing neointimal hyperplasia following arterial injury in vivo [see, Miller
et al.
(2000)]. Based on these findings, persons of skill accept that vitronectin
antagonists would be useful for the treatment of restenosis following PTCA.
Summary of the Invention
[0009] In one aspect the invention relates to compounds of formula I:
R6
O R2 R2a RR5
---T E L cIIic:7
R1 Y
R3 N \.._/
NHR8
I
3
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wherein
Y is chosen from the group consisting of -0-, -S-, -SO2-, -CH2- and
-N(loweralkyl)-;
L is a linker. The linker comprises from one to eight carbons and from
zero to three nitrogens, sulfurs and oxygens. At least two atoms are
interposed
between ring B and carbon P. The linker may be straight chain, branched or
cyclic
and, when cyclic, is either attached at carbons a and b of ring B or, when R'
is
methylene, it may be attached at Rl to form a four, five or six membered ring;
Q is chosen from 0, S, CH=N, N=CH, CH=CH and NR9;
E is hydroxy, or E is a biolabile residue such that E and the carboxyl to
which it is attached together form an ester or amide cleavable in vivo to
provide a
compound in which E is hydroxy;
R' is cliosen from the group consisting of hydrogen, aryl, heteroaryl, (C1
to C6)hydrocarbon, substituted aryl, Cl to C3 alkylaryl, -NHCOOR10,-NHSO2R10
and -NHCOR10; when R' is methylene (i.e. a divalent Cl hydrocarbon) it
provides
the point of attachment for a cyclic linker L;
R2 is chosen from the group consisting of hydrogen, aryl, heteroaryl, C1
to C6 hydrocarbon, substituted aryl, Cl to C3 alkylaryl, -NHCOOR10, -NHSO2R10
and NHCOR10, and Raa is hydrogen; or taken together RZ and Raa form a
carbonyl;
R3 and R4 are independently chosen from the group consisting of hydrogen, Ci
to
C4 hydrocarbon, loweralkoxy, halogen and fluoro(loweralkyl);
R5, R6 and R7 are independently chosen from the group consisting of hydrogen,
halogen and fluoro(loweralkyl);
R8 is chosen from hydrogen and lower alkyl; and
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R9 is chosen from hydrogen, alkyl, substituted alkyl, aryl and Cl to C3
alkylaryl; or
taken together R8 and R9 represent a two to four carbon chain forming a five
to
seven membered cyclic structure, which may contain one degree of unsaturation;
and
R10 is chosen from the group consisting of alkyl, substituted alkyl, aryl and
Cl to C3 alkylaryl.
[0010] In another aspect, the invention relates to a method of treating a
condition that is associated with excessive vitronectin receptor activity by
administering a therapeutically effective amount of a compound of the
foregoing
formula. Conditions associated with excessive vitronectin receptor activity
include: endometriosis, osteoporosis, restenosis following angioplasty,
rheumatoid
arthritis, cancer, macular degeneration and obesity.
Detailed Description of the Invention
[0011] The invention relates to compounds of formula I:
R6
O R R2a
R 4 Re R7
ay E L b
Ri Y
Q
3 N
NHR 8
I
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[0012] One embodiment includes subgenera in which L forms a fused ring with
ring B, such as the subgenus of formula:
O R R2a R's
E R 4 R R7
R1 0(
Y
Q
3 N \/
NHR8
in which L is a cyclic linker forming a five-, six or seven-membered ring. The
ring
formed by L may be substituted with one or two substituents, preferably lower
alkyl
and/or oxo. Particularly preferred compounds having cyclic L may be described
by
the subgeneric formula:
O R2 R2a R6
R4 R5 R7
E u ~. ~
1 I I
W Y
R3 N
NHR8
in which U is CH, C(CH3) or N; V is C=O, CHa or 0; W is (CH2)õC=O,
C(=O)(CH2)õ ,(CHZ)nCHa, O(CH2)õ or (CH2)õO; and n is zero, one or two.
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Examples of these include:
s
0 R2 R2a O R
R4 Rs R7
E N
1 I
R (CH2)p
Y
R3 N
NHR8
in which p is one, two or three;
Rs
0 R2 R2a R11
O R4 R5 R7
~
E R 1
I
~ ~ --~
Y
R3 N
NHR8
in which Rl l is hydrogen or methyl;
0 R2 Rs
R2a
R4 R5
E N
R1
/ ~ --~
O Y
R3 N
NHR$
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O R2 R6
R2a
R4 R5 R
E
1 I
R
Y
Rs N
NHR8
and
O R2 R6
R2a
O R4 R5 / R7
E
(
1
R ~ ~
O Y
R3 N
NHR$
Examples of cyclic L are:
8
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R R a
2
2 ~
2
R Me 0 T0-LLL / 'z,~
2 2
2
0
0
,-r 01 ~~.r ~
.
'~. 2 0~~
0
R R 0
~t
N 1 /, 1 R I-
2
~ R
.
r!'N
~-N
0 0 R R
0
p p Me p
Me
9
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[0013] Another embodiment includes subgenera in which L, by attaching
through R1, forms a ring incorporating the (3 carbon. These subgenera are
represented by the formula:
R6
0 R2 R2a R5
\ R /
E I I
6'H2
Q
R3 N \J
NHR$
[0014] A preferred subgenus is that of formula:
R6
0 R2 R2a 0
R4 Re R7
E N ~
I I
H2C (CH2)n
Q
R3 N '-)
NHR$
in which n is zero, one or two.
[0015] In another preferred embodiment, L may be an acyclic residue of one to
four carbons and from zero to three nitrogens, sulfurs and oxygens, in a
straight or
branched chain, para to Y:
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0 R2 R2a R 6
E R4 Rs / R7
R~ I
Y
Q
3 N \~
NHR 8
or from one to eight carbons and from zero to three nitrogens, sulfurs and
oxygens,
in a straight or branched chain naeta to Y:
R 6
4 R5 / R7
O R2 R2a
E
Ri R3 N
NHR$
Examples of acyclic L are CHaCH2, CH=CH, cyclopropane, amide, N(Loweralkyl)
amide, OCH2, sulfonamide, N(Loweralkyl) sulfonamide. Preferred compounds are
those in which L is -C(=O)NH-, -CH=CH- or -CH2CH2-
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[0016] Preferred subgenera of "Q" include compounds of formula:
R6
O R R2a 4 R~ 7
\ R /
E L
y Qa
3 N-_-
NHR$
in which Qa is 0, S, CH N, N=CH, CH=CH or NR9, and R9 is hydrogen, alkyl,
aryl, (C1-C3 )alkylaryl or alkyl substituted with methoxy, fluoro or hydroxy.
Particularly preferred ring systems are:
Rs
O R2
R4 E L LR5R7
R1
y
R3 N y
NHRB
R6
O R2
RR7
E L I I
R1 / \ 1
Y S
3
NHR8
and
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R6
O R R4 R5 RP
E
L I I
Y
R' y NR
Rs NHR 8
wherein R9 is chosen from hydrogen, lower alkyl, and fluoro(loweralkyl); and
R6
O R2
R4 R5 R7
E L I
R~
Y N
Rs N-'-
N--""'(CH2)m
/
R$
wherein m is one or two. A particularly preferred subgenus is the subgenus in
which a cyclic linker is combined with a cyclic structure formed from R8 and
R9:
2 R6
O R R2a
R4 R5 R7
E
R1
Y N
R$ N-
N- (CH2)m
/
R8
wherein m is one or two.
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An example of such a particularly preferred subgenus is:
O R2 R2a 0 R6
R4 R5 R7
E N
1 I (
Y N
R3
N -(CH2)m
~
R8
wherein m is one or two.
[0017] As described above, E may be hydroxy or a biolabile residue. The
active drug substance appears to be the carboxylic acid, i.e. E is hydroxy.
However, the invention also encompasses prodrugs of the active carboxylic
acids.
The concept of a prodrug is well established in the art (see for example US
patents
6,166,089; 5,681,964 and 4,235,887, the disclosures of which are incorporated
herein by reference). In the compounds that function as prodrugs, E and the
carboxyl to which it is attached together form an ester or amide cleavable in
vivo to
provide a compound in which E is hydroxy.
[0018] Turning to the two-carbon chain between the carboxylate and the linker
L, preferred compounds are those in which RZ is hydrogen, C1-C6 hydrocarbon,
aryl,
substituted aryl or heteroaryl; RZa is hydrogen and Rl is hydrogen, -
NHCOOR10,-NHCOR10 or -NHSO2R10 . When Rl is other than hydrogen, the
preferred stereochemistry of the carbon to which Rl is attached is of the
configuration shown:
O
E
R1 ~~N~
H
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[00191 When Rl is hydrogen, R2a is hydrogen and R2 is other than hydrogen, the
preferred stereochemistry of the carbon to which R2 is attached is of the
configuration shown:
O R 2 H
E
It is preferred that only one of Rl and R2 be other than hydrogen.
[0020] Among the ring substituents, preferred values for R3 and R4 are
hydrogen, methyl, methoxy, halogen or trifluoromethyl; for R5 and R7,
hydrogen;
for R8, hydrogen or methyl; and for Y, oxygen. The most preferred subgenus is
that
in which E is hydroxy; R' is hydrogen, -NHCOOR10 or -NHCOR10; Ra is hydrogen,
aryl, heteroaryl or substituted aryl; R3 and R4 are chosen from hydrogen,
methyl,
methoxy, halogen and trifluoromethyl; R5 and R7 are hydrogen; and R8 is chosen
from hydrogen and methyl.
[0021] The present invention includes compounds of formula (1) in the form of
salts. Suitable salts include those formed with both organic and inorganic
acids.
Such acid addition salts will normally be pharmaceutically acceptable,
although
salts of non-pharmaceutically acceptable salts may be of utility in the
preparation
and purification of the compound in question. The tenn "pharmaceutically
acceptable salt" refers to salts prepared from pharmaceutically acceptable non-
toxic
acids or bases including inorganic acids and bases and organic acids and
bases.
When the compounds of the present invention are basic, salts may be prepared
from
pharmaceutically acceptable non-toxic acids including inorganic and organic
acids.
Suitable pharmaceutically acceptable acid addition salts for the compounds of
the
present invention include acetic, benzenesulfonic (besylate), benzoic,
camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic, glutamic,
hydrobromic,
hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic,
mucic,
nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric acid, p-
toluenesulfonic, and the like. When the compounds contain an acidic side
chain,
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suitable pharmaceutically acceptable base addition salts for the compounds of
the
present invention include metallic salts made from aluminum, calcium, lithium,
magnesium, potassium, sodium and zinc or organic salts made from lysine, N,N'-
dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,
ethylenediamine, meglumine (N-methylglucamine) and procaine.
[0022] While it may be possible for the compounds of formula (I) or their
salts
and solvates to be administered as the raw chemical, it is preferable to
present them
as a pharmaceutical composition. According to a further aspect, the present
invention provides a pharmaceutical composition comprising a compound of
formula (I) or a pharmaceutically acceptable salt or solvate thereof, together
with
one or more pharmaceutically carriers thereof and optionally one or more other
therapeutic ingredients. The carrier(s) must be "acceptable" in the sense of
being
compatible with the other ingredients of the formulation and not deleterious
to the
recipient thereof.
[0023] The formulations include those suitable for oral, parenteral (including
subcutaneous, intradermal, intramuscular, intravenous and intraarticular),
rectal and
topical (including dermal, buccal, sublingual and intraocular) administration.
The
most suitable route may depend upon the condition and disorder of the
recipient.
The formulations may conveniently be presented in unit dosage form and may be
prepared by any of the methods well known in the art of pharmacy. All methods
include the step of bringing into association a compound of formula (I) or a
pharmaceutically acceptable salt or solvate thereof ("active ingredient") with
the
carrier which constitutes one or more accessory ingredients. In general, the
formulations are prepared by uniformly and intimately bringing into
association the
active ingredient with liquid carriers or finely divided solid carriers or
both and
then, if necessary, shaping the product into the desired formulation.
[0024] Formulations of the present invention suitable for oral administration
may be presented as discrete units such as capsules, cachets or tablets each
containing a predetermined amount of the active ingredient; as a powder or
granules; as a solution or a suspension in an aqueous liquid or a non-aqueous
liquid;
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or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The
active
ingredient may also be presented as a bolus, electuary or paste.
[0025] A tablet may be made by compression or molding, optionally with one
or more accessory ingredients. Compressed tablets may be prepared by
compressing in a suitable machine the active ingredient in a free-flowing form
such
as a powder or granules, optionally mixed with a binder, lubricant, inert
diluent,
lubricating, surface active or dispersing agent. Molded tablets may be made by
molding in a suitable machine a mixture of the powdered compound moistened
with
an inert liquid diluent. The tablets may optionally be coated or scored and
may be
formulated so as to provide sustained, delayed or controlled release of the
active
ingredient therein.
[0026] Formulations for parenteral administration include aqueous and non-
aqueous sterile inj ection solutions which may contain anti-oxidants, buffers,
bacteriostats and solutes which render the formulation isotonic with the blood
of the
intended recipient. Formulations for parenteral administration also include
aqueous
and non-aqueous sterile suspensions, which may include suspending agents and
thickening agents. The formulations may be presented in unit-dose of multi-
dose
containers, for example sealed ampoules and vials, and may be stored in a
freeze-
dried (lyophilized) condition requiring only the addition of a sterile liquid
carrier,
for example saline, phosphate-buffered saline (PBS) or the like, immediately
prior
to use. Extemporaneous injection solutions and suspensions may be prepared
from
sterile powders, granules and tablets of the kind previously described.
[0027] Formulations for rectal administration may be presented as a
suppository
with the usual carriers, such as cocoa butter or polyethylene glycol.
[0028] Formulations for topical administration in the mouth, for example
buccally or sublingually, include lozenges comprising the active ingredient in
a
flavored basis such as sucrose and acacia or tragacanth, and pastilles
comprising the
active ingredient in a basis such as gelatin and glycerin or sucrose and
acacia.
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[0029] Preferred unit dosage formulations are those containing an effective
dose, or an appropriate fraction thereof, of the active ingredient.
[0030] The pharmaceutical compositions will usually include a
"pharmaceutically acceptable inert carrier" and this expression is intended to
include one or more inert excipients, which include starches, polyols,
granulating
agents, microcrystalline cellulose, diluents, lubricants, binders,
disintegrating
agents, and the like. If desired, tablet dosages of the disclosed compositions
may be
coated by standard aqueous or nonaqueous techniques. "Pharmaceutically
acceptable carrier" also encompasses controlled release means. Compositions of
the present invention may also optionally include other therapeutic
ingredients,
anti-caking agents, preservatives, sweetening agents, colorants, flavors,
desiccants,
plasticizers, dyes, and the like.
[0031] The compounds of formula (1) are preferably administered orally or by
injection (intravenous or subcutaneous). The precise amount of compound
administered to a patient will be the responsibility of the attendant
physician.
However, the dose employed will depend on a number of factors, including the
age
and sex of the patient, the precise disorder being treated, and its severity.
Also, the
route of administration may vary depending on the condition and its severity.
Definitions
[0032] Throughout this specification the terms and substituents retain their
definitions.
[0033] Alkyl is intended to include linear, branched, or cyclic hydrocarbon
structures and combinations thereof. Lower alkyl refers to alkyl groups of
from 1
to 6 carbon atoms. Examples of lower alkyl groups include methyl, ethyl,
propyl,
isopropyl, butyl, s-and t-butyl and the like. Preferred alkyl groups are those
of C20
or below. Cycloalkyl is a subset of alkyl and includes cyclic hydrocarbon
groups
of from 3 to 8 carbon atoms. Examples of cycloalkyl groups include c-propyl, c-
butyl, c-pentyl, norbomyl and the like.
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[0034] (C1 to Cn )Hydrocarbon includes alkyl, cycloalkyl, alkenyl, alkynyl,
aryl
and combinations thereof containing only hydrogen and one to n carbons.
Examples include vinyl, allyl, cyclopropyl, propargyl, phenethyl,
cyclohexylmethyl, camphoryl and naphthylethyl.
[0035] Alkoxy or alkoxyl refers to groups of from 1 to 8 carbon atoms of a
straight, branched, cyclic configuration and combinations thereof attached to
the
parent structure through an oxygen. Examples include methoxy, ethoxy, propoxy,
isopropoxy, cyclopropyloxy, cyclohexyloxy and the like. Lower-alkoxy refers to
groups containing one to four carbons.
[0036] Oxaalkyl refers to alkyl residues in which one or more carbons (and
their associated hydrogens) have been replaced by oxygen. Examples include
methoxypropoxy, 3,6,9-trioxadecyl and the like. The term oxaalkyl is intended
as it
is understood in the art [see Naming and Indexing of Chemical Substances for
Chemical Abstracts, published by the American Chemical Society, 196, but
without the restriction of 1127(a)], i.e. it refers to compounds in which the
oxygen
is bonded via a single bond to its adjacent atoms (forming ether bonds); it
does not
refer to doubly bonded oxygen, as would be found in carbonyl groups.
[0037] Acyl refers to groups of from 1 to 8 carbon atoms of a straight,
branched, cyclic configuration, saturated, unsaturated and aromatic and
combinations thereof, attached to the parent structure through an carbonyl
functionality. One or more carbons in the acyl residue may be replaced by
nitrogen, oxygen or sulfur as long as the point of attachment to the parent
remains
at the carbonyl. Examples include acetyl, benzoyl, propionyl, isobutyryl, t-
butoxycarbonyl, benzyloxycarbonyl and the like. Lower-acyl refers to groups
containing one to four carbons.
[0038] Aryl and heteroaryl mean a 5- or 6-membered aromatic or
heteroaromatic ring containing 0-3 heteroatoms selected from 0, N, or S; a
bicyclic
9- or 10-membered aromatic or heteroaromatic ring system containing 0-3
heteroatoms selected from 0, N, or S; or a tricyclic 13- or 14-membered
aromatic
or heteroaromatic ring system containing 0-3 heteroatoms selected from 0, N,
or S.
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The aromatic 6- to 14-membered carbocyclic rings include, e.g., benzene,
naphthalene, indane, tetralin, and fluorene and the 5- to 10-membered aromatic
heterocyclic rings include, e.g., imidazole, pyridine, indole, thiophene,
benzopyranone, thiazole, furan, benzimidazole, quinoli.ne, isoquinoline,
quinoxaline, pyrimidine, pyrazine, tetrazole and pyrazole.
[0039] Alkylaryl means an alkyl residue attached to an aryl ring. Examples of
Cl-C3 alkylaryl are benzyl, phenethyl, phenylpropyl and naphthylethyl.
Alkylheteroaryl means an alkyl residue attached to a heteroaryl ring. Examples
include, e.g., pyridinylmethyl, pyrimidinylethyl and the like.
[0040] Heterocycle means a cycloalkyl or aryl residue in which from one to
three carbons is replaced by a heteroatom selected from the group consisting
of N,
0 and S. The nitrogen and sulfur heteroatoms may optionally be oxidized, and
the
nitrogen heteroatom may optionally be quaternized. Examples of heterocycles
that
fall within the scope of the invention include pyrrolidine, pyrazole, pyrrole,
indole,
quinoline, isoquinoline, tetrahydroisoquinoline, benzofuran, benzodioxan,
benzodioxole (commonly referred to as methylenedioxyphenyl, when occurring as
a
substituent), tetrazole, morpholine, thiazole, pyridine, pyridazine,
pyrimidine,
thiophene, furan, oxazole, oxazoline, isoxazole, dioxane, tetrahydrofuran and
the
like. It is to be noted that heteroaryl is a subset of heterocycle in which
the
heterocycle is aromatic. Examples of heterocyclyl residues additionally
include
piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxo-pyrrolidinyl, 2-
oxoazepinyl,
azepinyl, 4-piperidinyl, pyrazolidinyl, imidazolyl, imidazolinyl,
imidazolidinyl,
pyrazinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolyl,
quinuclidinyl,
isothiazolidinyl, benzimidazolyl, thiadiazolyl, benzopyranyl, benzothiazolyl,
tetrahydrofuryl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl,
thiamorpholinylsulfoxide, thiamorpholinylsulfone, oxadiazolyl, triazolyl and
tetrahydroquinolinyl.
[0041] Substituted alkyl, aryl, cycloalkyl, heterocyclyl etc. refer to alkyl,
aryl,
cycloalkyl, or.heterocyclyl wherein up to three H atoms in each residue are
replaced
with halogen, haloalkyl, hydroxy, loweralkoxy including alkylene dioxy,
carboxy,
CA 02611971 2007-12-12
WO 2007/001249 PCT/US2005/020656
carboalkoxy (also referred to as alkoxycarbonyl), carboxarnido (also referred
to as
alkylaminocarbonyl), cyano, carbonyl, nitro, amino, alkylamino, dialkylamino,
mercapto, alkylthio, sulfoxide, sulfone, acylamino, amidino, phenyl, benzyl,
heteroaryl, phenoxy, benzyloxy, or heteroaryloxy.
j0042] The term "halogen" means fluorine, chlorine, bromine or iodine.
[0043] The term "oxo", when referring to a substituent, means an oxygen
double bonded to a carbon, e.g. a ketone or amide carbonyl.
[0044] As used herein, reference to "treatment" or "treating" a patient are
intended to include prophylaxis. The terms include amelioration, prevention
and
relief from the symptoms and/or effects associated with these disorders. The
terms
"preventing" or "prevention" refer to administering a medicament beforehand to
forestall or obtund an attack. Persons of ordinary skill in the medical art
(to which
the present method claims are directed) recognize that the term "prevent" is
not an
absolute term. In the medical art it is understood to refer to the
prophylactic
administration of a drug to diminish the likelihood or seriousness of a
condition,
and this is the sense intended.
[0045] The graphic representations of racemic, ambiscalemic and scalemic or
enantiomerically pure compounds used herein are taken from Maehr J. Chem. Ed.
62, 114-120 (1985): solid and broken wedges are used to denote the absolute
configuration of a chiral element; wavy lines indicate disavowal of any
stereochemical implication which the bond it represents could generate; solid
and
broken bold lines are geometric descriptors indicating the relative
configuration
shown but denoting racemic character; and wedge outlines and dotted or broken
lines denote enantiomerically pure compounds of indeterminate absolute
configuration. Thus, among the structures below, those having open wedges are
intended to encompass both of the pure enantiomers of that pair, those having
solid
wedges are intended to encompass the single, pure enantiomer having the
absolute
stereochemistry shown.
[0046] Abbreviations
21
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[0047] The following abbreviations and terms have the indicated meanings
throughout:
Ac = acetyl
ACN = acetonitrile
Bn = benzyl
BNB = 4-bromomethyl-3-nitrobenzoic acid
Boc = t-butyloxy carbonyl
Bu = butyl
CBZ = carbobenzoxy = benzyloxycarbonyl
DBU = diazabicyclo[5.4.0]undec-7-ene
DCM = dichloromethane = methylene chloride = CHaCIa
DEAD = diethyl azodicarboxylate
DIC = diisopropylcarbodiimide
DIEA = N,N-diisopropylethyl amine
DIPEA = N,N-diisopropylethyl amine
DMAP = 4-N,N-dimethylaminopyridine
DME = dimethoxyethane
DMF = N,N-dimethylformamide
DMSO = dimethyl sulfoxide
DVB = 1,4-divinylbenzene
EDC = ethylene dichloride = 1,2-dichloroethane
EEDQ = 2-ethoxy-l-ethoxycarbonyl-1,2-dihydroquinoline
Et = ethyl
FCC = flash column chromography
Fmoc = 9-fluorenylmethoxycarbonyl
GC = gas chromatography
HATU = O-(7-Azabenzotriazol=l-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate
HOAc = acetic acid
HOBt = hydroxybenzotriazole
m-CPBA = m-chloroperbenzoic acid
Me = methyl
22
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WO 2007/001249 PCT/US2005/020656
mesyl = methanesulfonyl
MTBE = methyl t-butyl ether
NMM = N-methylmorpholine
NMO = N-methylmorpholine oxide
NMP = N-methylpyrrolidone
PEG = polyethylene glycol
Ph or x = phenyl
PhOH = phenol
Pfl' = pentafluorophenol
PPTS = pyridinium p-toluenesulfonate
PyBroP = bromo-tris-pyrrolidino-phosphonium hexafluorophosphate
rt = room temperature
sat'd = saturated
s- = secondary
t- = tertiary
TBAB = tetrabutylammonium bromide
TBAI = tetrabutylammonium iodide
TBDMS = t-butyldimethylsilyl
TEA = triethylamine
TFA = trifluoroacetic acid
THF = tetrahydrofuran
TMOF = trimethyl orthoformate
TMS = trimethylsilyl
Ts = tosyl = p-toluenesulfonyl
Trt = triphenylmethyl
[0048) Although this invention is susceptible to embodiment in many different
forms, preferred embodiments of the invention are shown. It should be
understood,
however, that the present disclosure is to be considered as an exemplification
of the
principles of this invention and is not intended to limit the invention to the
embodiments illustrated.
23
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[0049] It may be found upon examination that certain members of the claimed
genus are not patentable to the inventors in this application. In this event,
subsequent exclusions of species from the compass of applicants' claims are to
be
considered artifacts of patent prosecution and not reflective of the
inventors'
concept or description of their invention; the invention encompasses all of
the
members of the genus (I) that are not already in the possession of the public.
[0050] In general, the compounds of the present invention may be prepared by
the methods illustrated in the general reaction schemes as, for example,
described
below, or by modifications thereof, using readily available starting
materials,
reagents and conventional synthesis procedures. In these reactions, it is also
possible to make use of variants that are in themselves known, but are not
mentioned here.
24
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WO 2007/001249 PCT/US2005/020656
Preparation of In.termediates 1-5 and 2-5:
O F qF Cs2CO3 0
'_O R NOa '_O R
OH DMF, 80 C 0
R R NOZ
R = OMe: 99% X
R = OMe or Me R = Me: 96% F
1-1 R=OMe
2-1 R=Me
0
CH3NH2,DIPEA __O ~ R I~ ~ 10% Pd/C, H2 (50psi)
DMF, r.t., 2 da s l~ O N~
Y I anhydrous EtOH
R NO2 H
R = OMe: 100% R = OMe: 70%
R"-Me: 80% R= Me: 100%
1-2 R=OMe
2-2 R=Me
O 0
~Ojt, q R 13rCN, MeOH ON r.t. I s O I? N--
R NH2 H R N~
R = OMe: 76% NH2
R = Me: 99%
1-3 R=OMe 1-4 R=OMe
2-3 R=Me 2-4 R=Me
0
LiOH, THF/MeOH/H20(6/3/1) HO I~ R I
or NaOH, EtOH b / /
N-
reflux R Nzz<
R = OMe: 47% NH2
R = Me: 65% 1-5 R=OMe
2-5 R=Me
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Preparation of Intennediate 1-5:
Step l:
[0051] To a solution of methylsyringate (1.0g; 4.72 mmol) in DMF (50 mL)
were added CsZCO3 (1.85 g; 5.66 mmol) and 2,6-difluoronitrobezene (0.5 mL;
4.72
mrnol). The reaction mixture was heated to 80 C for 16 h. The mixture was
cooled and poured into water, then extracted with EtOAc (3X). The organic
layers
were combined and washed with water (1X) then brine (1X). The organic phase
was dried (MgSO4), filtered and concentrated in vacuo giving 1.64 g (99%) of 1-
1
which was used in the next reaction without ftirther purification.
Step 2:
[0052] To a solution of 1-1 (1.46g, 4.16 mmol) in DMF (25 mL) were added
methylamine (2M in MeOH, 9.4 mL, 18.9 mmol) and diisopropylethylamine (1.64
mL; 9.44 mmol). The mixture was stirred at room temperature for 48 h. The
mixture was then poured into water and extracted with EtOAc (3X). The combined
organic layer was washed with brine (1X) and dried over MgSO4, filtered and
concentrated in vacuo. The residue obtained was chromatographed on silica gel
with n-hexane-EtOAc (1:1) to give 1.69 g of 1-2 (100%) as a yellowish solid.
Step 3:
[0053] A solution of 1-2 (1.69g, 4.66 mmol) in anhydrous ethanol (50mL)
containing 10 % of palladium on activated carbon (-50 mg) was shaken under a
hydrogen atmosphere (50 psi) in a Parr apparatus for 16 h. The mixture was
filtered
through the celite and washed with ethanol. The filtrate was concentrated in
vacuo
to yield 1.08 g (70%) of 1-3 which was used in the next step without further
purification.
Data for 1-3: MS: na/z (assignment, relative intensity) 333.0 (M+H+, 80).
26
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Step 4:
[0054] To a solution of 1-3 (1.08 g, 3.25 mmol) in anhydrous methanol under
argon, was added 5.0 M cyanogen bromide in acetonitrile (2.60 mL, 13.0 mmol).
The mixture was stirred for 16 h at room temperature and then conoentrated in
vacuo. The residue was chromatographed on silica gel with CH2C12-CH3OH (95:5)
giving 880 mg (76%) of 1-4.
Data for 1-4: MS: m/z (assignment, relative iintensity) 358.3 (M+W, 65).
Step 5:
[0055] A mixture of 1-4 (880 mg, 2.46 mmol) and lithium 12ydroxide hydrate
(540 mg, 13.00 mmol) in THF/MeOH/H20 (6:3:1) (40 mL) was refluxed for 16h.
The nizxture was concentrated in vacuo to dryness. The residue was re-
dissolved in
distilled water and the solution obtained was adjusted to pH around 5 with 1N
HC1
(aq). Precipitated solid was collected by filtration and dried in vacuo to
give 400 mg
(47%) of 1-5. Data for 1-5: MS: m/z (assignment, relative intensity) 344.2
(M+H+, 60).
Preparation of Intermediate 2-5:
Ste~~1:
[0056] Following the general procedure described for step 1 in example 1, 4-
hydroxy-3,5-dimethyl-benzoic acid methyl ester (1.0 g, 5.55 mmol) was reacted
with 2,6-difluronitrobenzene giving 1.70 g (96%) of crude 2-1 which was used
in
the next step without purification.
StM 2:
[0057] Following the general procedure described for step 2 in example 1, the
crude compound 2-1 was treated with methylamine to give 1.40 g (80 %) of 2-2
after FCC purification (silica gel, n-Hexane / EtOAc 3:1).
27
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Step 3:
[00581 Following the general procedure described for step 3 in example 1, the
nitro compound 2-2 was reduced by hydrogenation yielding 1.29 g(100 J ) of
corresponding amine 2-3 which was used directly in the next step without
further
purification.
Ste .~}4:
[0059] Following the procedure described for step 4 in example 1, compound 2-
3 was reacted with cyanogen bromide to yield, after purification by FCC
(silica gel,
eluted with 5% methanol/DCM), 1.38 g (99%) of aminobenzimidazole 2-4.
Data for 2-4: MS: m/z (assig11111ent, relative intensity) 326.1 (M+H}, 100).
St~
[0060] The compound 2-4 obtained above (1.38 g, 4.25 mmol) was combined
with 1N NaOH (aq) (10.6 mL, 10.6 mmol) in 20 mL EtOH. After 6 h at 60 C the
mixture was cooled and 10.6 mL 1 N HCl (aq) was added, adjusted fmal pH around
5. The resulting white precipitate was collected on a frit, washed with water,
then
with EtZO and dried providing 870 mg (65%) of acid 2-5. Data for 2-5: MS: m/z
(assignment, relative intensity) 312.3 (M+H+, 100).
28
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WO 2007/001249 PCT/US2005/020656
Exam.pl e 3:
~-o 0
o I ~
o o o o
HO / O~ H2N N
~ O~
H
~ ~
~ I ~ I
00 O N- N~ HOBt, EDC, Et3N, r.t. O\N N
~ oO --'
1_5 NH2 15% NH2
O--\ 3-1
O
1 N NaOH (aq), MeOH, 70 C O O N , O
100% HO H
00 N--~
NH2
3
step1:
[0061] To a solution of 1-5 (20 mg; 0.0583 mmol) in DMF (1 mL) were added
triethylamine (10 L; 0.064 mmol), 1-hydroxybenzotriazole (9 mg; 0.064 mmol),
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (12 mg; 0.064
mmol) and (S)-3-Amino-3-benzo[1,3]dioxol-5-yl-propionic acid ethyl ester
hydrochloride (16 mg; 0.0583 mmol) [For preparation see: Zablocki, J. A.; et
al. J.
Med. Chein. 1995, 38, 2378. ] The reaction mixture was stirred at room
temperature overnight. Then the mixture was partitioned between EtOAc and
water. The aqueous phase was extracted with EtOAc (3X). The organic phases
were combined and concentrated in vacuo. The residue was purified by FCC
(silica
gel; elution with 1:1 EtOAc :Hexane) giving 5 mg (15%) of 3-1.
Data for 3-1: MS: m/z (assignment, relative intensity) 563.1 (M+H+, 100).
29
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WO 2007/001249 PCT/US2005/020656
Step 2:
[0062] To a solution of ethyl ester 3-1 (5 mg; 0.0089 mmol) in MeOH (2 mL)
was added 1 N NaOH (aq) (50 L; 0.050 mmol). The resultant mixture was heated
to 70 C for 16h. The mixture was then cooled and concentrated in vacuo and
the
crude material was acidified to pH 2 with 1 N HC1(aq). The mixture was applied
to a column filled with Dowex 50 W (H) ion exchange resin and then eluted with
water until the eluent became neutral (pH 6). The column was then eluted with
5%
pyridine/H20. Fraction containing the desired product were pooled and
concentrated in vacuo and the zwitterion thus obtained was treated with 50%
TFA/DCM and concentrated in vacuo giving 6 mg of 3 (100%) as the TFA salt
Data for 3: MS: m/z (assignment, relative intensity) 535.2 (M+H-'-, 100).
Exainple 4:
0II
O NH2 O O
HO NHCBz ~O~H / I O~~
\ O\ N~ OuNH \ O\ N-
HOBt, EDC, Et3N, r.t. I O N=~
O I
O
NH2 78% 4-1 NH2
1-5 \ I
o O
50%TFA/DCM HO/Jj-_ '_\N /
Oy NH H \ I O\ N~
O O N~
70% ~ NH2
4
Step1:
[00631 To a solution of 1-5 (20 mg; 0.0583 mmol) in DMF (0.8 mL) were
added triethylamine (9.47 L; 0.07 mmol), 1-hydroxybenzotriazole (9.45 mg; 0.07
mmol), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (13.4 mg;
0.07 mmol) and (S)-3-amino-2-benzyloxycarbonylamino-propionic acid tert-butyl
ester (7.14 mg; 0.0583 mmol) [For preparation see: Stilz, Hans Ulrich; et al.
J.
CA 02611971 2007-12-12
WO 2007/001249 PCT/US2005/020656
Med. Cliem. 2001, 44, 1158-1176.] The reaction mixture was stirred at room
temperature overnight. Then the mixture was partitioned between EtOAc and
water. The aqueous phase was extracted with EtOAc (3X). The organic phases
were combined and concentrated in vacuo. The residue was purified by FCC
(silica
gel; elution with 1:1 EtOAc :Hexane) giving 28 mg (78%) of 4-1.
Data for 4-1: MS: m/z (assigmnent, relative intensity) 620.2 (M+H}, 100),
564.2
(M-t-Bu, 67).
Step ?:
[0064] To a solution of 4-1 (28 mg; 0.0533 mmol) in DCM (3 mL) was added
trifluoroacetic acid (3 mL). The reaction mixture was stirred at room
temperature
for 4 hours. The mixture was then concentrated in vacuo and the residue was
triturated with ethyl ether (3X) giving 18 mg (70%) of 4. Data for 4: MS: m/z
(assignment, relative intensity) 564.2 (M+H+, 100).
31
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WO 2007/001249 PCT/US2005/020656
Example 5:
I~ 4-fluorobenzonitriie, NC 1 N NaOH (aq), reflux;
\ N~ Cs2CO3, DMF, 80 C ~ I O~ I HCI (aq)
OH N\
O 5-1
O O
HO / I i I :z::::z THF, I ester H
N N\
5-2 5-3
O O
m-CPBA, DCM, ~O~Nkao TsCI, DCM; NH4OH (aq)
0 Cto23 C H <
5-4 O N ~ I
O ' ~ O
~O~H ~' 4 N HCI/dioxane HO H ~ I
~ O ~ O
NI N-
5-5 NHa NH2
*HCI
st~
[0065] To a solution of 8-hydroxyquinoline (1g; 6.89 mmol) in DMF (7 mL)
were added Cs2CO3 (2.24 g; 6.89 mmol) and 4-fluorobenzonitrile (834 mg; 6.89
mmol). The reaction mixture was heated to 80 C for 16 h. The mixture was
cooled and diluted with EtOAc (300 mL). This was washed with water (2 X 100
niL) and brine (1 X 100 mL). The organic phase was dried (NaaSO4), filtered
and
concentrated in vacuo giving 1.32 g(78%) of 5-1 which was used in the next
reaction without f.urther purification.
Step 2:
[0066] Benzonitrile 5-1 (1.32 g; 5.36 mmol) was treated with 13 mL of 1 N
NaOH (aq) and heated to reflux for 22 h. The mixture was then cooled to room
32
CA 02611971 2007-12-12
WO 2007/001249 PCT/US2005/020656
temperature and 13 mL of 1 N HC1(aq) was added carefully forming a tan
precipitate which was collected by filtration and washed with cold water (2X)
to
afford 1.39 g of 5-2 (98%).
Data for 5-2: MS: m/z (assignment, relative intensity) 266.2 (M+IT'", 100).
Step 3:
[0067] To a solution of 5-2 (197 mg; 0.743 mmol) in THF (7 mL) at 0 C were
added i-butylchloroformate (96 L; 0.743 mmol) and NMM (90 L; 0.817 mmol)
and the resultant mixture was stirred at 0 C for 90 s. To this were added (3-
alanine
t-butyl ester = HCl (135 mg; 0.743 mmol) and TEA (114 L; 0.817 mmol). The
mixture was allowed to warm to room temperature and stirred for 0.5 h after
which
it was filtered through a pad of Celite and the filtrate diluted with EtOAc
(100 mL).
This was washed with sat. Na2CO3 (aq) (1 X 50 mL) and brine (1 X 50 mL), dried
(Na2SO4), filtered and concentrated in vacuo. The residue was purified by FCC
(silica gel; elution with 2:1 EtOAc:hexanes) giving 234 mg (80%) of 5-3.
Data for 5-3: MS: fn/z (assignment, relative intensity) 393.0 (M+H-'-, 100),
337.1
(M-tBu, 21).
StM4:
[0068] To a solution of quinoline 5-3 (48 mg; 0.122 mmol) in DCM (2 mL) at 0
C was added in-CPBA (70%; 30 mg, 0.122 mmol). The reaction mixture was
allowed to warm to room temperature and stirred for 3 d. The mixture was then
filtered through basic alumina (elution with DCM then 5% MeOH/DCM) to remove
any unreacted m-CPBA and the filtrate was concentrated in vacuo. The crude
residue was then purified by FCC (silica gel; elution with 5% MeOH/DCM) to
afford 29 mg (58%) of N-oxide 5-4.
St~
[0069] To a solution of the N-oxide 5-4 (29 mg; 0.0710 mmol) in DCM (0.5
mL) was added TsCl (16 mg; 0.0852 mmol) and this was immediately followed by
the addition of conc. NH40H (aq) (0.15 mL). The biphasic mixture was stirred
vigorously at room temperature for 3 h and was then diluted with EtOAc and
33
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WO 2007/001249 PCT/US2005/020656
washed with sat. NaHCO3 (aq) (1X). The organic phase was dried (Na2SO4),
filtered and concentrated in vacuo and the residue was purified by FCC (silica
gel;
elution with 5% MeOH/DCM) giving 9 mg (31%) of aniinoquinoline 5-5.
Data for 5-5: MS: nz/z (assignment, relative intensity) 408.0 (M+H+, 100),
352.2
(M-tBu, 13).
Step6:
[0070] 5-5 was treated with 4 N HCl in dioxane (2 mL) and allowed to stand for
16 h. This was then concentrated in vacuo giving 9 mg of 5 as an HCl salt.
Data for 5: MS: fn/z (assignment, relative intensity) 352.2 (M+H'', 100).
34
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Example 6:
0 HO O acetic anhydride MOjL, q O isobutylene, H2SO4(cat) IOH DMAP, DCM O
CHCI3:dioxane(3:1)
,-O ~-O
100% 94%
6-1
O \
0
N
~O I \ ~
0 K2C03 ~O \ O Br
)OW / O MeOH OH
(CH3CN)4CuPF6
98% Cs2CO3, pyridine, 100 C
6-2 6-3 47%
O O 0
p O 1) TFA/DCM "~v~ _
ON O
EDCRDMAP, DCM k H O\
~O N~ 00 N 6-4 6-5
0 0 mCPBA /jjv_'H I\ O/ 1) tosyl chloride
DCM /X\ / O\ I 2)DCM H
50% O,N
6-6 35%
O O O 0
ko TFA/DCM HO~H ~\OO/ ~
O / \
~O N~ ~O N~ I
NHa
70% NH2
6-7
6
Step 1:
[0071] To syringic acid (9.9g, 50mmo1) in lOOmL DCM was added DIVIAP
(6.7g, 55mmol). To this solution was added slowly acetic anhydride (5.6g,
55mmol). The solution was stirred for 3 hours and then concentrated in vacuo.
The
CA 02611971 2007-12-12
WO 2007/001249 PCT/US2005/020656
residue was taken up in DCM and washed with 1N HCl. The organic layer was
dried over MgSO4 and concentrated to yield 6-1 as an orange solid (12g,
50mmo1,
100%).
Step 2
[0072] To 6-1 (1.3 g, 5.4mmo1)in 30 mL of CHC13:dioxane (3:1) in a pressure
vessel at -78 C was added approx l OmL isobutylene and a few drops of
concentrated sulfuric acid. The vessel was sealed, wanned to 25 C for 2 hours,
then heated to 80 C for 18 hours. The solution was then concentrated in vacuo.
The residue was taken up in DCM and washed with water. The organic layer was
dried over MgSO4 to yield 6-2 as a white solid (1.5g, 5.lmrnol, 94%).
Steb3
[00731 To 6-2 (1.5g, 5.lmmol) in 30mL MeOH was added K2C03 (1.4g,
l Ommol) and stirred for 1 hour. The mixture was filtered and concentrated in
vacuo. The residue was taken up in DCM and washed with 1N HCI. The organic
layer was dried over MgSO4 to yield 6-3 as an oil (1.3g, 5mmol, 98%).
Sten 4
[0074] 6-3 (0.25g, lmmol), 8-bromoquinoline (0.21g, lmmol), Cs2CO3 (1.0g,
3mmol), and tetrakisacetonitrile copper(I)hexafluoroacetate (0.37g, lmmol)
were
suspended in 2.5m1 anhydrous pyridine under an Argon atmosphere and heated at
100 C. After 8 hours and additional 0.37g of and tetrakisacetonitrile
copper(I)hexafluoroacetate was added. After 18 hours the mixture was diluted
with
water and extracted with DCM. The organic layer was concentrated in vacuo and
the residue chromatagraphed on a silica gel column, eluting with 30%
EtOAc:hexanes to yield 6-4 as a foam (0.18g, 0.47mmo1, 47%).
Data for 6-4: MS: in/z (assignment, relative intensity) 326.3 (M+H+ -tbu, 90)
Step5
[0075] 6-4 (0.3g, 0.8mmol) was treated with lOml 50% TFA/DCM for 45
minutes. The solution was concentrated in vacuo. The residue was taken up in
36
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DCM and basified with excess DIEA. To this solution was added N-
methylmorpholine (290 1, 2.6mmol) and isobuytylchloroformate (115 1, 0.9mmo1).
After 10 minutes (3-alanine t-butyl ester (0.18g, 0.96mmol) was added. The
solution was stirred for 18 hours. The solution was then concentrated in
vacuo.
The residue was taken up in DCM and washed with 0.5N HCl and 0.5N NaOH.
The organic layer was dried over MgSO4 and concentrated. The resulting oi16-5
was used without purification.
Data for 6-5: MS: na/z (assignment, relative intensity) 453.3 (M+H+, 90)
Step 6
[0076] To 6-5 in lOml DCM was added 3-perchlorobenzoic acid (0.69g,
4mmol). The mixture was stirred for 18 hours then diluted with DCM and washed
with 1N NaOH. The organic layer was dried over MgSO4 and concentrated to yield
6-6 as a glass (0.1g, 0.2 mmol, 50% over 2 steps)
Step 7
[0077] To 6-6 (0.1g, 0.2mmol) in 1.8m1 CHC13 was added tosyl chloride (0.5g,
0.26mmo1) immediately followed by ammonium hydroxide (110 1, 2m1/mmol).
The solution was stirred for 90 minutes then diluted with saturated NaHCO3 and
extracted with CHC13. After concentration, the residue was chromatagraphed on
a
silica gel column eluting with 5%MeOH/DCM to 7%MeOH/DCM to yield 6-7 as a
foam (0.033g, 0.07mmol, 35%).
Data for 6-7: MS: m/z (assignment, relative intensity) 468.2 (M+H+, 95)
Step 8
[0078] 6-7 was treated with l Oml TFA/DCM for 6 hours. The solution was
then concentrated and purified by preparative HPLC to yield 6 as a tan solid
(0.02g,
0.05im1iol, 70%).
Data for 6: MS: m/z (assignment, relative intensity) 412.6 (M+W, 90)
37
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Example 7:
O
Br acetic anhydride, TEA, Br IX_IL.
OH DMAP, DCM 4-Bromo-2,6-dimethylphenol 93% 7-1
ethyl-4-pentenoate, Pd(OAc)2, P(o-tol)3, O 0 K2C03, MeOH, 2h
/ / I O
TEA, ACN, reflux, 16h O 92%
87% 7-2
0 2,6-difluoronitrobenaene, Cs2CO3,
O DMF, 80 C
OH
7-3 34%
0 MeNH2, DIPEA, DMF 0
'-O / + O / ~
O~ F 57% O~ N
NO2 NO2 H
7-4 7-5
0
SnCl2*2H20, EtOH, reflux BrCN, MeOH
45% O y- 40%
NH2
7-6
0 0
O / \ I \ ' LiOH, MeOH/THF/H20 HO \ ' \ I
O N- (3:3:1) O
N=-{ N~
NH2 NH2
7-7
7
st~
[0079] To a solution of 4-bromo-2,6-dimethylphenol (1.00 g; 4.97 nmmol) in
DCM (25 mL) were added triethylamine (1.04 mL; 7.46 mmol) and acetic
anhydride (0.56 mL; 5.96 mmol). To this was added a small scoop of DMAP and
38
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the reaction mixture was stirred at 23 C for 1 h. The volatiles were removed
in
vacuo and the residue was taken up in EtOAc (100 mL). The organic solution was
washed with 1 M HCl (aq) (1 X 40 mL), sat. NaHCO3 (aq) (1 X 40 mL) and brine
(1 X 40 mL). The organic phase was dried (Na2SO4), filtered and concentrated
in
vacuo giving 1.13 g of 7-1 (93%) which was used in the next step without
further
purification.
Step 2:
[0080] To a solution of 7-1 (500 mg; 2.06 mmol) and ethyl-4-pentenoate (0.26
mL; 2.06 mmol) in acetonitrile (10 mL) were added TEA (0.57 mL; 4.12 mmol),
P(o-tol)3 (82 mg; 0.268 mmol) and Pd(OAc)2 (46 mg; 0.206 mmol). The reaction
vessel was flushed with Argon and the reaction mixture was then heated to
reflux
for 16 h. The mixture was cooled, concentrated in vacuo and the residue was
purified by FCC (silica gel; elution with 5:1 hexanes:EtOAc) affording 520 mg
of
7-2 (87%).
Step 3:
[0081] To a solution of 7-2 (520 mg; 1.80 mmol) in MeOH (9 mL) was added
potassium carbonate (371 mg; 2.69 mmol) and the mixture was stirred at 23 C
for
2 h. The solvent was then removed in vacuo and the crude residue was
partitioned
between EtOAc and sat. NH4C1(aq). The organic phase was washed with brine
(1X), dried (Na2SO4), filtered and concentrated in vacuo to afford 390 mg of
phenol
7-3 (92%) which was used in the following reaction without further
purification.
Ste4:
[0082] To a solution of phenol 7-3 (0.39 g; 1.66 mmol) in DMF (16 mL) were
added 2,6-difluoronitrobenzene (0.18 mL; 1.66 mmol) and Cs2CO3 (541 mg; 1.66
mmol). The reaction mixture was heated to 80 C for 16 h. The mixture was
cooled, diluted with EtOAc (100 mL) and washed with water (1 X 50 mL) and
brine (1 X 50 mL). The organic phase was dried over Na2SO4, filtered and
concentrated in vacuo and the crude residue was purified by FCC (silica gel;
elution
with 5:1 hexanes:EtOAc) giving 210 mg of 7-4 (34%).
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Sto 5:
[0083] To a solution of 7-4 (210 mg; 0.562 mmol) in DMF (5 mL) were added
MeNH2 (2.0 M in THF; 0.56 mL; 1.12 mmol) and DIPEA (0.2 mL; 1.12 mmol).
The resultant mixture was stirred at 23 C for 16 h. More MeNH2 (2.0 M in THF;
0.56 mL; 1.12 mmol) was added and stirring was continued for another 16 h.
After
diluting with EtOAc (50 mL) the mixture was washed with water (1 X 20 mL) and
brine (1 X 20 mL). The organic phase was dried (NaaSO4), filtered and
concentrated in vacuo. The residue was purified by FCC (silica gel; elution
with
2:1 hexanes:EtOAc) giving 123 mg of 7-5 (57%).
Data for 7-5: MS: m/z (assignment, relative intensity) 385.0 (M+H+, 100).
Step 6:
[0084] To a solution of alkene 7-5 (123 mg; 0.320 mmol) in EtOH (3 mL) was
added SnC12.2Ha0 (361 mg; 1.60 mmol). The reaction mixture was heated to
reflux
for 16 h. The mixture was then cooled and concentrated in vacuo. Water was
added (10 mL) and the pH was adjusted to 9 with sat. NaHCO3 (aq). This was
extracted with EtOAc (3 X 20 mL) and the combined organics were washed with
water (1 X 20 mL) and brine (1 X 20 mL). The organic layer was then dried
(NaaSO4), filtered and concentrated in vacuo giving 53 mg of ethyl ester 7-6
(45%).
Data for 7-6: MS: m/z (assignment, relative intensity) 369.1 (M+H+, 100).
Ste7:
[0085] To a solution of diamine 7-6 (53 mg; 0.144 mmol) in MeOH (1 mL) was
added cyanogen bromide (0.5 M in acetonitrile; 0.12 mL; 0.575 mmol). The
reaction mixture was stirred at 23 C for 3 days. The reaction was quenched
with
sat. NaHCO3 (aq) and concentrated in vacuo. The crude residue was partitioned
between EtOAc and water. The organic phase was washed with brine (1X), dried
.(Na2SO4), filtered and concentrated in vacuo. Purification by FCC (silica
gel;
elution with 10% MeOH/DCM) afforded 23 mg (40%) of aminobenzimidazole 7-7.
Data for 7-7: MS: m/z (assignment, relative intensity) 394.3 (M+W, 100).
CA 02611971 2007-12-12
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step 8:
[0086] To a solution of the ethyl ester 7-7 (23 mg; 0.0585 mmol) in MeOH (3
mL), THF (3 mL) and water (1 mL) was added LiOH=H2O (20 mg; XS). The
reaction mixture was stirred at 23 C for 16 h. The volatiles were removed in
vacuo
and the mixture was acidified to pH 1 with 2 N HCl (aq). The aqueous solution
was
applied to a column filled with Dowex 50 W(FIF) resin. The column was washed
with water until the eluant became neutral and was then eluted with 5%
pyridine/H20. The desired fractions were combined and concentrated in vacuo.
The crude material thus obtained was then purified by FCC (silica gel; elution
with
8:2:0.2 DCM:MeOH:H20) giving 7.5 mg (36%) of acid 7.
Data for 7: MS: m/z (assignment, relative intensity) 366.3 (M+H+, 100).
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Exam ine8:
p
~ ~ Ha O / I BBr3, DCM,
p I xO v'NHZ o
/ p ~--~ p -78 C to 23 C;
I TEA, MeOH, reflux +O MeOH
Br
43% 8-1
O 2,6-difluoronitrobenzene, O
O N ta s2 s O N ~ ~ ?
__/- OH O F
-p -O NO2
8-2 47% for 2 steps 8-3
O
1) H2 (9; 1 atm), 10% Pd/C,
methylamine, DMF, 23 C p---N EtOH
taO
N 2) BrCN, MeOH
100% NOa
8-4 28% for 2 steps
O O LiOH, MeOH/THF/H20 (3:2:1)
-ON 5zII:Li~:I1
O N' 58%
N-=~
NH2
8-5
O O
HO~N
O N~
N=-~
NH2
8
step 1:
[0087] To a solution of 2-Bromomethyl-4-methoxy-benzoic acid methyl ester
(310 mg;
1.20 mmol) [For preparation see: Wyrick, S. D.; et al. J. Med. Chem. 1987, 30,
1798-1806 and references therein] in MeOH (2 mL) were added triethylamine (418
L; 3.00 mmol) and (3-alanine t-Bu ester hydrochloride (240 mg; 1.32 mmol). The
reaction mixture was heated to reflux for 3 h. The mixture was then cooled,
42
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concentrated in vacuo and the crude residue was taken up in EtOAc. This was
washed with 1 N HCl (aq) (1X), dried (NaaSO4), filtered and concentrated in
vacuo.
The residue was purified by FCC (silica gel; elution with 1:1 EtOAc:hexanes)
giving 152 mg (43%) of 8-1.
Data for 8-1: MS: na/z (assignment, relative intensity) 291.9 (M+H+, 100),
236.1
(M-tBu, 25%).
StM 2:
[0088] To a solution of 8-1 (152 mg; 0.522 mmol) in DCM (2 mL) at -78 C
was added BBr3 (IM in DCM; 1.8 mL; 1.8 mmol). The mixture was allowed to
warm to room temperature and stirred for 16 h. The mixture was then cooled to -
78 C and quenched with MeOH (1 mL). The mixture was allowed to warm to
room temperature and the volatiles were removed in vacuo. The crude residue
was
azeotroped with MeOH (3X) and used in the next reaction without further
purification.
Data for 8-2: MS: m/z (assignment, relative intensity) 236.1 (M+H+, 100).
St~
[0089] To a solution of 8-2 (123 mg; 0.522 mmol) in DMF (5 mL) were added
2,6-difluoronitrobenzene (55 L; 0.522 mm.ol) and Cs2CO3 (170 mg; 0.522 mmol).
The reaction mixture was heated to 80 C for 16 h. The mixture was cooled and
partitioned between EtOAc and water. The aqueous phase was extracted with
EtOAc (3X). The organic phases were combined and washed with water (1X) and
brine (IX). The organic phase was dried (Na2SO4), filtered and concentrated in
vacuo. The residue was purified by FCC (silica gel, elution with 3:1
EtOAc:hexanes) giving 91 mg (47% for 2 steps) of 8-3.
Stgp 4:
[0090] To a solution of 8-3 (91 mg; 0.243 mmol) in DMF (2 mL) was added
MeNH2 (2.0 M in MeOH; 0.49 mL; 0.972 mmol). The reaction mixture was stirred
at room temperature for 16 h. The mixture was diluted with EtOAc (100 mL) and
43
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washed with water (1 X 25 mL) and brine (1 X 25 mL). The organic phase was
dried (Na2SO4), filtered and concentrated in vacuo. The desired aniline 8-4
was
used crude in the next reaction.
Step 5:
(0091] 8-4 (95 mg; 0.243 mmol) was reduced by hydrogenation in EtOH in the
presence of a catalytic amount of 10% Pd/C under 1 atm of H2 (g) (balloon) for
6
h. The reaction mixture was filtered through a pad of Celite and concentrated
in
vacuo. The crude residue was taken up in MeOH (2 mL) and to the resultant
solution was added cyanogen bromide (5 M in acetonitrile; 0.19 mL; 0.972
mmol).
The mixture was stirred for 16 h at room temperature and was then quenched
with
sat. NaHCO3 (aq) and concentrated in vacuo. The residue was partitioned
between
EtOAc and water and the aqueous phase was extracted with additional EtOAc
(2X).
The organic phases were combined, dried (Na2SO4), filtered and concentrated in
vacuo.
The residue was purified by FCC (silica gel, elution with 10% MeOH/DCM)
affording 26 mg of the desired aminobenzimidazole 8-5.
Data for 8-5: MS: m/z (assignment, relative intensity) 381.3 (M+W, 100), 761.0
(2M + H+, 6).
StM 6:
j0092] To a solution of methyl ester 8-5 (26 mg; 0.0683 mmol) in MeOH (1.5
mL), THF (1 mL) and water (0.5 mL) was added LiOH=H20 (9 mg; 0.205 mmol).
The reaction mixture was stirred for 0.5 h and then more LiOH=H2O (8 mg) was
added after which stirring was continued for 2 h. The mixture was then
concentrated in vacuo and the residue treated with 1 mL of water. This was
brought
to pH 4-5 by addition of 1 N HCl (aq) giving an off-white precipitate which
was
collected by filtration. The solid thus obtained was treated with 50% TFA/DCM
and concentrated in vacuo affording 19 mg of 8 as a TFA salt.
Data for 8: MS: na/z (assignment, relative intensity) 367.2 (M+H+, 100), 733.1
(2M + H}, 22).
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Example 9:
~O I\ LiOH, THF/MeOH/H20 HO~~,~~
0
(3:3:1) O
96% 9-1
1) (COCI)2, DCM
2) TMSdiazomethane, THF/ACN (1:1), O
0 C Bn BBr3, DCM,
~O \
3) benzyl alcohol, collidine, 180 C ~, -78 C-> 23 C; MeOH
O 1~
9-2
87% for 3 steps
O 2,6-difluoronitrobenzene, 0
~ \ CsCO3, DMF, 80 C ( \ I \
~ OH / O ~ .F
9-3 52% for 2 steps 9-4 NO2
0
\ ~ H2 (g; 1 atm), 10% Pd/C,
methylamine, DMF, 23 C '-O EtOH
I~ol/N
84% NO2 H 11 %
9-5
0 0
BrCN, MeOH "~p I \
( / I a i
O N
i O N-
NHZ H 50% N--~
9-6 9-7 NH2
0
1 N NaOH (aq), MeOH, 40 C; HO \ ~
H+ + / O I ~ N~
N=~ * TFA
85% NHz
9
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Example 9:
Step 1:
[0093] To a solution of (6-Methoxy-1,2,3,4-tetrahydro-naphthalen-2-yl)-acetic
acid ethyl ester (307 mg; 1.13 mmol) [For preparation see: Fisher, M. J.; et
al. J.
Med. Chem 1999, 42, 4875-4889] in THF (3 mL), MeOH (3 mL) and water (1 mL)
was added LiOH-HaO (142 mg; 3.38 mmol). The reaction mixture was stirred at
room temperature for 2 hours and was then concentrated in vacuo. The residue
was
made acidic by the addition of 1 N HCl (aq) and the cloudy white mixture was
extracted with EtOAc (3X). The combined organic layers were dried (NaZSO4),
filtered and then concentrated in vacuo giving 264 mg of acid 9-1 (96%) which
was
of suitable purity for synthetic use in the next step.
Step 2:
[0094] To a solution of acid 9-1 (83 mg; 0.340 mmol) in DCM (1 mL) were
added oxalyl choride (36 L; 0.408 mmol) and a drop of DMF (cat.). The
reaction
mixture was stirred at room temperature for 0.5 h until bubbling ceased. The
mixture was concentrated in vacuo and then azeotroped with toluene (2x). The
crude acid chloride was dissolved in THF (0.5 mL) and ACN (0.5 mL), cooled to
0 C, and to this was added TMSdiazomethane (2.0 M in hexanes; 0.37 mL; 0.748
mmol). The reaction mixture was stirred at 0 C for 2.5 h and was then
concentrated in vacuo. Benzyl alcohol (0.5 mL) and collidine (0.5 mL) were
then
added to the crude diazoketone and the mixture was heated to 180 C for 10
min.
The mixture was cooled to room temperature, diluted with ethyl ether and
washed
with 1N HCl (aq) (1 X 20 mL), water (1 X 10 mL) and brine (1 X 10 mL). The
ether layer was dried (Na2SO4), filtered and concentrated in vacuo. The crude
product was purified by FCC (silica gel; elution with 6:1 hexaries:EtOAc)
giving
99mg of benzyl ester 9-2(87%).
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Step 3:
[0095] Following the general procedure described for step 2 in example 8, the
product from above, 9-2, was reacted with BBr3 giving phenol 9-3 which was
used
in the next step crude without further purification.
Step 4:
[0096] Following the procedure described for step 3 in example 8, the crude
product from above, 9-3, was reacted with 2,6-difluoronitrobenzene giving,
after
purification by FCC (silica gel; elution with 4:1 hexanes : EtOAc), 67 mg (64%
for
2 steps) of 9-4.
Step 5:
[0097] Following the procedure described for step 4 in example 8, compound 9-
4 from above was reacted with methylamine to afford 58 mg (84%) of 9-5 after
purification by FCC (silica gel; elution with 4:1 hexanes:EtOAc).
Step 6:
[0098] 9-5 (58 mg; 0.151 mmol) was reduced by hydrogenation in EtOH in the
presence of a catalytic amount of 10% Pd/C under 1 atm of H2 (g) (balloon) for
16
h. The reaction mixture was filtered through a pad of Celite and concentrated
in
vacuo. The residue was purified by FCC (silica gel, elution with 2:1
hexanes:EtOAc) to afford 6 mg (11%) of the desired diamine 9-6.
Data for 9-6: MS: m/z (assignrnent, relative intensity) 355.3 (M+H'', 100).
Step 7:
[0099] The diamine 9-6 above was taken up in MeOH (1 mL) and to the
resultant solution was added cyanogen bromide (5 M in ACN; 0.014 mL; 0.0677
mmol). The mixture was stirred for 1 h at room temperature and was then
quenched with sat. NaHCO3 (aq) and concentrated in vacuo. The residue was
partitioned between EtOAc and brine and the aqueous phase was extracted with
additional EtOAc (2X). The organic phases were combined, dried (Na2SO4),
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filtered and concentrated in vacuo. The residue was purified by FCC (silica
gel,
elution with 10% MeOH/DCM) affording 3 mg (50%) of the desired
aminobenzimidazole 9-7.
Data for 9-7: MS: m/z (assignment, relative intensity) 380.5 (M+W, 100).
Step 8:
(00100] To a solution of methyl ester 9-7 (3 mg; 0.00791 mmol) in MeOH (0.5
mL) was added 50 L of 1 N NaOH (aq). The resultant mixture was stirred at
room
temperature for 16 h. Analysis by HPLC showed incomplete reaction so more 1 N
NaOH (aq) was added (50 L) and the mixture was heated to 40 C for lh. The
mixture was then cooled and concentrated in vacuo and the crude material was
acidified to pH 2 with 1 N HCl (aq). The crude suspension was applied to a
column
filled with Dowex 50 W(H) ion exchange resin and then eluted with water until
the eluent became neutral (pH 6). The column was then eluted with 5%
pyridine/H20. Fractions containing the desired product were pooled and
concentrated in vacuo and the zwitterion thus obtained was treated with 50%
TFA/DCM and concentrated in vacuo giving 3.4 mg of 9 (85%) as the TFA salt.
Data for 9: MS: m/z (assignment, relative intensity) 366.5 (M+H+, 100)
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Example 10:
0
0 ~-O OH 4-oxo-butyric acid te-f-butyl ester,
, pyrrolidine, MeOH, reflux OH
HOI ~
78% 0
10-1
0
1) NaBH4, MeOH ~ 2,6-difluoronitrobenzene,
O Cs2CO3, DMF, 80 C
2) H2 (g) (50psi), Pd(OH)2/C, EtOH I~ OH
10-2 23% for 3 steps
4 O ~ , methylamine, DMF, 23 C
O
0
I~ O\ I F 62%
NO2
10-3
1) Ha (g; 1 atm), 10% Pd/C,
EtOH
0
I/ O\ I F 2) BrCN, MeOH,
NOa
10-4 25% for 2 steps
0
-O O I~ / I 50% TFA/DCM
O
N=~
NH2
10-5
0
HO
N \
NH2 *TFA
Step 1:
[00101] To a solution of 4-oxo-butyric acid tert-butyl ester (53 mg; 0.335
mmol)
in MeOH (3 mL) were added 2',5'- dihydroxyacetophenone (51 mg; 0.335 mmol)
and pyrrolidine (28 L; 0.335 mmol). The reaction mixture was heated to 70 C
for
49
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16 h. The mixture was cooled, concentrated in vacuo and the residue was
purified
by FCC (silica gel; elution with 3:1 hexanes:EtOAc) to afford 76 mg (78%) of
10-1.
StM2:
[00102] To a solution of 10-1 (76 mg; 0.260 mmol) in MeOH (5 mL) was added
NaBH4 (12 mg; 0.312 mmol). The reaction mixture was stirred for 1 h and was
then concentrated in vacuo. The residue was treated with brine and extracted
with
EtOAc (3x). The organic layers were combined, dried (Na2SO4), filtered and
concentrated in vacuo. The crude alcohol was taken up in EtOH and the
resulting
solution placed in a Parr shaker vessel. To this was added a small scoop of
Pd(OH)2/C. The mixture was placed under 50 psi of H2 (g) and shaken for 16 h
in a
Parr hydrogenator. The mixture was then filtered through a pad of Celite and
concentrated in vacuo giving 72 mg (100%) of 10-2 which was used without
further
purification.
Data for 10-2: MS: :n/z (assignment, relative intensity) 301.0 (M+Na+, 100),
222.9
(M-tBu, 42).
Step3:
[00103] Following the procedure described for step 3 in example 8, the crude
product from above, 10-2, was reacted with 2;6-difluoronitrobenzene giving,
after
purification by FCC (silica gel; elution with 4:1 hexanes:EtOAc), 25 mg (23%
for 3
steps) of 10-3.
Sten 4:
[00104] Following the procedure described for step 4 in example 8, compound
10-3 from above was reacted with methylamine to afford 16 mg (62%) of 10-4
after
purification by FCC (silica gel; elution with 4:1 hexanes:EtOAc).
Stg 5:
[00105] Following the procedure described for step 5 in example 8, compound
10-4 from above was reduced and then reacted with cyanogen bromide to afford,
CA 02611971 2007-12-12
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after purification by FCC (silica gel; elution with 10% MeOH/DCM), 4 mg (25%
for 2 steps) of aminobenzimidazole 10-5.
Data for 10-5: MS: m/z (assignment, relative intensity) 424.1 (M+W, 100),
368.3
(M-tBu, 72).
Step 6:
[00106] 10-5 (4mg; 0.0094 mmol) was treated with 50% TFA/DCM and allowed
to stand for 16 h. This was then concentrated in vacuo and triturated with
Et20
giving 4 mg (89%) of 10 as a TFA salt.
Data for 10: MS: tn/z (assignment, relative intensity) 368.2 (M+H+, 100).
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Example 11:
0
NaN3 benzyl bromide
HN
OH TFA, 4 OH K2CO3, DMF
98% 11-1 36%
O 0 0
HN I \ ethyl bromopropionate ~O)~N \
/ O (\ NaH, DMF I/ O \
11-2 67%
11-3
H2, Pd/C O O nI,,,
F F
~
EtOH NO2
100% H Cs2CO3, DMF
11-4 70%
O 0 0 0
O--\/-~ao,P,F CH3NH2 ~O/ v 'N \ /
DIEA, DMF I/ O\ H
NO2 NO2 30% 11-6
11-5
0 0
Zn, AcOH /-~O)\~N \ / CNBr
THF I/ O\ I Ns EtOH
100% NH2 H 36%
11-7
0 0 0 0
1N LiOH J~'~
/\O~N \ THF HO N I\ / I
i I / \
/ O N~ p
N~ 38% N-
11-8 NH2 NH2
11
st~
[00107] To a solution of 5-hydroxyindanone (1.5g, 10 minol) in 15m1
trifluroracetic acid was added sodium azide (1.0g, 15 mmol) portionwise. The
solution was stirred at reflux for 2 h, then cooled and diluted with water.
The
volatiles were removed in vacuo and the resulting residue diluted with
saturated
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NaHCO3 and extracted with ethyl acetate. The organic layers were dried over
MgSO4 and concentrated to yield 11-1 (1.6g, 98%) as a tan solid which was used
without further purification.
Step2:
[00108] To a solution of 11-1 (0.8g, 5mmo1) in 10 ml DMF was added K2C03
(1.4g, lOmmol) and benzyl bromide (1.7g, lOmmol). The mixture was stirred at
25 C for 18 h. The resulting mixture was then filtered and concentrated in
vacuo.
The resulting oil was purified on a silica gel column, eluting with 5%
MeOH/DCM
to yield 11-2 (0.46g, 1.8mmo1, 3 6%) as an oil.
Sten 3:
[00109] A solution of 11-2 (0.46g, 1.8mmo1) in 15m1 anhydrous DMF was
added to NaH (60% in mineral oil, 110mg, 2.7mmol) slowly. The resulting
mixture was stirred at 25 C for 2 h. A solution of ethyl bromopropionate
(0.39g,
2.2mmo1) in 5m1 anhydrous DMF was then added dropwise. The mixture was
stirred for 18 h. The mixture was slowly diluted with l Oml water. The
solution
was concentrated in vacuo and the residue purified on a silica gel column,
eluting
with 2% MeOH/DCM to yield 11-3 ( 0.42g, 1.2mmo1, 67%) as an oil.
StM4:
[00110] To a solution of 11-3 ( 0.42g, 1.2mmo1) in lOml EtOH was added a
small scoop of 10% Pd/C. The mixture was stirred at 25 C under a H2 atmosphere
for 18 h. The mixture was then filtered and concentrated in vacuo to yield 11-
4 as
an oil (0.32g, 1.2mmol, 100%)
Data for 11-4: MS: m/z (assignment, relative intensity) 264.1 (M+H+, 95)
Step 5:
[00111] A solution of 11-4 (0.32g, 1.2mmol), 2,6-difluoronitrobenzene (0.23g,
1.4mmo1), and Cs2CO3 (0.86g, 2.6mmo1) in lOml DMF was stirred at 60 C for 5
hours. The mixture was cooled to room temp, filtered and concentrated in
vacuo.
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The residue was chromatagraphed on a silica gel column, eluting with 5%
MeOH/DCM to yield 11-5 as an oil (0.34g, 0.84mmol, 70%).
Step 6:
[00112] To 11-5 (0.32g, 0.84mmol) in 5m1 DMF was added DIEA (435 l,
2.5mmol) and methylamine (lml 2M in THF). The solution was stirred at 25 C for
18 h. The solution was concentrated in vacuo and the resulting oil was
chromatagraphed on a silica gel column, eluting with 5% MeOH/DCM to yield 11-
6 as an oil (0.21g, 0.50mmol, 60%).
Step 7:
[00113] To a solution of 11-6 (0.21g, 0.50mmo1) in lOml THF was added lml
acetic acid and 0.50g of Zn dust. The mixture was stirred at reflux for 30
min. The
mixture was filtered and concentrated in vacuo to yield 11-7 as an oil which
was
used without further purification.
St~8:
[00114] To 11-7 (0.21g, 0.50mmol) in 5ml EtOH was added cyanogen bromide
(l.Oml; 5M in acetonitrile) and stirred at 25 C for 2 h. The solution was
concentrated in vacuo and the resulting oil chromatagraphed on a silica gel
column,
eluting with 5% MeOH/DCM to 7% MeOH/DCM to yield 11-8 as a foam (0.07g,
0.18mmo1, 36%).
Step 9:
[00115] To 11-8 (0.07g, 0.18mmol) in 5m1 THF was added 200 l of 1N LiOH
(aq) and stirred at 25 C for 18 h. The mixture was concentrated in vacuo and
the
residue taken up in 7ml water and treated with 200 1 of 1N HCl (aq). The
precipitate was filtered and then treated with 4N HCUdioxane. Concentration in
vacuo yielded 11 as a pale orange solid (28 mg, 0.07mmol, 3 8%).
Data for 11: MS: m/z (assignment, relative intensity) 381.3 (M+H+, 90).
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Examnle 12:
O benzyl bromide O
O O NaBH4
OH K2C03, DMF ~ O \ -~
MeOH
/- s0 ~ /
55% for
12-1 2 steps
O
I\ SOC12 O KCN, 18-C-6 (cat.)
HO O ~ CI (s \O
CH3CN:H20 (1:1)
,O / O
12-2 81% 33%
12-3
\ O ~
O
NC I / O \ BH3"TH '
THF H2N O
1-O
12-4 82% 12-5
ethyl chtoroformate O \
'I ~ ethyl bromopropionate
DIEA, DCM ~O, N / O NaH, DMF
H s0 I i
44%
12-6 75%
O O triflic anhydride 0 0 '---O-'-N O DMAP, DCM O
~a ~
O
12-7 12-8
0 0 1
H2, Pd/C ,/,0)~,~N O
10-
EtOH OH
67% for i0
2 steps 12-9
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~ 1
O O
I I ~ ~
J~ p / O N
~O N ~/ F NOa F~ I/ O~ I F
OH
0 Cs2CO3, DMF 1-1O NO2
i
12-9 10% 12-10
O O ~
~J p Zn, AcOH
CH3NH2 9 T-
DIEA, DMF 0 N
/0 NOZ
12-11
O (
0
O O 1 CNBr qoo
p
/
N ~ \ N~
O ~ N~ Et0
s0 N=~
,O NH2 H 25% NH2
12-13
12-12
O O
IN NaOH ~~
THF
HO O N-
(!q-q
36% ,O N=~
NH2
12
stepl:
[00116] 4-hydroxy-2,5-dimethoxybenzaldehyde (5.0g, 27.4mmo1) in 75m1 DMF
was added Cs2CO3 (18g, 54.9 mmol) and benzyl bromide (6.5ml, 54.9mmol). The
mixture was heated at 75 C for 1 h. The mixture was filtered and concentrated
in
vacuo. The resulting yellow solid 12-1 (7.9g) was used without further
purification.
Step 2:
[00117] To a solution of 12-1 in 75m1 MeOH was added sodium borohydride
(1.9g, 50mmo1) portionwise. The resulting mixture was stirred for 18 h. The
mixture was then concentrated in vacuo. The residue was taken up in DCM and
56
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washed with 0.5N HCl. The organic layer was dried over MgSO4 and concentrated
to yield 12-2 as a yellow solid. (5.5g, 15mmol, 55% over 2 steps).
Step3:
[00118] To 12-2 (1.5g, 5.5mmol) was added 20m1 of thionyl chloride and heated
at reflux for 30 min. The solution was concentrated in vacuo and the residue
taken
up in DCM and washed with water. The organic layer was dried over MgSO4 and
concentrated to yield 12-3 as an oil. (1.3g, 4.4mmol, 81%)
Step 4:
[00119] To 12-3 (1.0g, 3.4mmol) in 20m1 CH3CN:water (1:1) was added
potassium cyanide (0.36g, 5.5mmol) and a crystal of 18-crown-6. The solution
was
stirred 18 h. The solution was then concentrated in vacuo and chromatagraphed
on
a silica gel column, eluting with EtOAc:hexanes (1:1) to yield 12-4 as a solid
(0.3g,
1.lmmol, 33%).
Step 5:
[00120] To 12-4 (0.3g, l.lmmol) in 5m1 anhydrous THF was added BH3-THF
(2ml l.OM in THF) and stirred for 18 h. The solution was then carefully
treated
with HCUEtOH (5m1, 9%W/V). After 2 h the solution was concentrated in vacuo.
The residue was taken up in DCM and washed with 1N NaOH. The organic layer
was dried over Na2SO4 and concentrated to yield 12-5 as an oil. (0.25g,
0.9mmol,
82%).
Data for 12-5: MS: nz/z (assignment, relative intensity) 288.1 (M+H+, 80)
Step 6:
[00121] To sodium hydride (70mg, 1.8mmo160% dispersion in oil) under argon
was added 12-5 (0.25g, 0.9mmol) in 5m1 anhydrous DMF dropwise. After 2 h ethyl
3-bromopropionate (0.18g, 1.35mmo1) in 5ml anhydrous DMF was added
dropwise. The mixture was stirred for 18 h after which 5m1 iN HCl was added
slowly. The solution was then concentrated in vacuo. The residue was
57
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chromatagraphed on a silica gel column, eluting with 2% MeOH/DCM to yield 12-
6 as an oil. (155mg, 0.4mmol, 44%)
Step 7:
[00122] To 12-6 (155mg, 0.4mmol)in 5m1 DCM was added DIEA (200m1,
l.lmmol) and ethyl chloroformate (100 m11.0 mmol). The solution was stirred
for
2 h then concentrated in vacuo. The residue was taken up in DCM and washed
with
1N NaOH, 1N HCI, and brine. The organic layers were dried over Na2SO4 and
concentrated to yield 12-7 as an oil (150mg, 0.3mmol, 75%)
Step 8:
[00123] To trifluoromethane sulfonic anhydride (0.2ml, 1.5mmo1) in 2.5m1
anhydrous DCM cooled to 0 C was added a solution of 12-7 (150mg, 0.3mmol) and
DMAP (110mg, 0.9mmol) in 12m1 anhydrous DCM dropwise. The solution was
stirred at 0 C for 18 h. The solution was then quenched with water and
extracted.
The organic layer was concentrated in vacuo and chromatagraphed on a silica
gel
column, eluting with 5% MeOH/DCM. The product 12-8 was then taken up in 5m1
EtOH and a small scoop of 10% Pd/C was added. This mixture was placed under a
balloon of hydrogen for 18 h. The mixture was then filtered and concentrated
in
vacuo to yield 12-9 as an oil (75mg, .2mmol, 67% for 2 steps)
Step 9: .
[00124] Following the general procedure described for example 11, step 5, 12-9
was reacted with 2,6-difluoronitrobenzene affording after purification on a
silica gel
column, eluting with 25% EtOAc/hexanes 10 mg of 12-10 (10%).
Step 10:
[00125] Following the general procedure described for example 11, step 6, 12-
10
was treated with methylamine to afford 12-11. The crude product was used
without
purification.
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Steb11:
[00126] Following the general procedure described for example 11, step 7, 12-
11
was treated with Zn and acetic acid in THF to afford 12-12. The crude product
was
used without purification.
Step 12:
[00127j Following the general procedure described for example 11, step 8, 12-
12
was treated with cyanogen bromide to afford 12-13 which was chromatagraphed on
a silica gel column, elution with 5% MeOH/DCM, to afford 2.5mg (25% for 3
steps).
Data for 12-13: MS: na/z (assignment, relative intensity) 468.5 (M+H'', 95)
Step13:
[00128] Following the general procedure described for example 11, step 9, 12-
13
was treated with IN NaOH to afford 12 which was purified by preparative HI'LC
giving 0.8mg (36%).
Data for 12: MS: m/z (assignment, relative intensity) 441.2 (M+H+, 95)
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Example 13:
c(NH2 1) TMSCI, Et3N, THF N 2) n-BuLi, THF, -78 C; j
trans-ethylcinnamate
OMe OMe
100% 13-1
~
ethylchloroformate, Et3N, O Tf2O, DMAP, DCM, 0 C
N
DCM O 15% (33% b.o.r.s.m.)
26% OMe
13-2
\ \
~ / ~ /
O O BBr3, DCM, -78 C-> 23 C; O O
~O u N / EtOH N / ~
'' I O~ \ OH
13-3 13-4
2,6-difluoronitrobenzene, O O methylamine, DMF, 23 C
CsaCO3, DMF, 80 C O U N / I
\ O 89%
83% for 2 steps NOZ
13-5
\
0 I/ O 1) H2 (9; 1 atm), O O
10% Pd/C, EtOH
2) BrCN, MeOH N \ I
O N O
NO2 H 59% for 2 steps
13-7 NH2
13-6
f \
/
1 N NaOH (aq), MeOH, 70 C; O O
H+ HO N ao
/ ~ \
N-
N=~ *TFA
13 NH2
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stm 1:
[00129] To a solution of 3-methoxyphenethylamine (0.5 mL; 3.41 mmol) and
triethylainine (0.68 mL; 4.88 mmol) in THF (3 mL) was added TMSCI (0.51 mL;
4.02 mmol). The reaction mixture was stirred for 1 h at room temperature. To
the
resultant thick slurry was added more THF (3 mL) and then this was filtered
through a sintered glass funnel into a 25 mL r.b. flask under a blanket of
argon.
The resulting clear solution was cooled to -78 C and n-BuLi (2.5 M; 1 mL;
2.56
mmol) was added and stirred for 15 min at the same temperature. Trans-
ethylcinnamate (0.29 mL; 1.71 mmol) was added via cannula as a solution in THF
(1 mL) and the mixture stirred for an additional 15 min at -78 C before
quenching
with sat. ammonium chloride (aq). The mixture was warmed to room temperature
and 1 N HCl (aq) was added forming a thick white precipitate. The mixture was
basified to pH 10-11 with 1 N NaOH (aq) and extracted with DCM (3X). The
combined organic layers were dried (Na2SO4), filtered and concentrated in
vacuo.
The crude residue was purified by FCC (silica gel, elution with 2:1
hexanes:EtOAc)
giving 560 mg (100%) of 13-1.
Data for 13-1: MS: m/z (assignment, relative intensity) 328.1 (M+W, 100).
Step 2:
[00130] To a solution of 13-1 (610 mg; 1.86 mmol) in DCM (10 mL) at room
temperature were added triethylamine (0.39 mL; 2.79 mmol) and
ethylchloroformate (0.21 mL; 2.24 mmol). A crystal of DMAP was added and the
reaction mixture was stirred at room temperature for 1 h afterwhich more
triethylamine (0.39 mL; 2.79 mmol) and ethylchloroformate (0.21 mL; 2.24 mmol)
were added. After continuing to stir at room temperature for 3 d, the mixture
was
concentrated in vacuo and the crude residue was taken up in EtOAc. This was
washed with 1 N HCl (aq) (1 X 20 mL), sat. NaHCO3 (aq) (1 X 20 mL) and brine
(1 X 20 mL). The organic phase was dried (NazSO4), filtered and concentrated
in
vacuo. The crude product was purified by FCC (silica gel, elution with 3:1
hexanes: EtOAc) to afford 190 mg (26%) of ethyl carbamate 13-2.
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Step 3:
[00131] To a solution of 13-2 (190 mg; 0.48 mmol) and DMAP (183 mg; 1.50
mmol) in DCM (20 mL) at 0 C was added dropwise triflic anhydride (0.40 mL;
2.40 mmol) as a solution in DCM (4 mL). The resultant cloudy mixture was
stirred
for an additional 2 h at 0 C and was then diluted with ether (40 mL) and
washed
with sat. NaHCO3 (aq) (1X) and 1 N HCl (aq) (1X). The organic phase was dried
(Na2SO4), filtered and concentrated in vacuo. The crude mixture was purified
by
FCC (silica gel, elution with 2:1 hexanes:EtOAc) giving 26 mg (15%) of
isoquinolinone 13-3 along with 100 mg of recovered starting material 13-2 (33%
based on reacted starting material).
St~
[00132] Following the general procedure described for step 2 in example 8, the
product from above, 13-3 (35 mg; .0991 mmol), was reacted with BBr3 giving
pheno113-4 which was used in the next step crude without farther purification.
Data for 13-4: MS: m/z (assignment, relative intensity) 340.0 (M+H'-, 100).
Step 5:
[00133] Following the procedure described for step 3 in example 8, the crude
product from above, 13-4, was reacted with 2,6-difluoronitrobenzene giving,
after
purification by FCC (silica gel; elution with 2:1 hexanes:EtOAc), 39 mg (83%
for 2
steps) of 13-5.
Step 6:
[00134] Following the procedure described for step 4 in example 8, compound
13-5 from above was reacted with methylamine to afford 34 mg (89%) of 13-6
after
purification by FCC (silica gel; elution with 2:1 hexanes:EtOAc).
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Sten 7:
[00135] Following the procedure described for step 5 in example 8, compound
13-6 from above was reduced and then reacted with cyanogen bromide to afford,
after purification by FCC (silica gel; elution with 5% MeOH/DCM then 10%
MeOHIDCM), 20 mg (59% for 2 steps) of aminobenzimidazole 13-7.
Data for 13-7: MS: nz/z (assignment, relative intensity) 485.2 (M+H+, 100),
969.0
(2M+H+, 32).
Stm 8:
[001361 To a solution of ethyl ester 13-7 (20 mg; 0.0413 mmol) in MeOH (2
mL) was added 1 N NaOH (aq) (0.12 mL; 0.12 mmol). The resultant mixture was
heated to 70 C for 3h. The mixture was then cooled and concentrated in vacuo
and
the crude material was acidified to pH 2 with 1 N HCl (aq). The mixture was
applied to a column filled with Dowex 50 W(H) ion exchange resin and then
eluted with water until the eluent became neutral (pH 6). The column was then
eluted with 5% pyridine/H20. Fraction containing the desired product were
pooled
and concentrated in vacuo and the zwitterion thus obtained was treated with
50%
TFA/DCM and concentrated in vacuo giving 8.3 mg of 13 (33%) as the TFA salt.
Data for 13: MS: m/z (assignment, relative intensity) 457.2 (M+H+, 100), 913.1
(2M+W, 30).
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Example14:
\
O FI~ F 0
'-O / NOZ O / ethanolamine, MeOH, 80 C
OH Cs2CO3, DMF O\ I F sealed tube, 16h
100% NO2
14-1 63%
0
~1 O / 1) H2 (g; 1 atm), 10% Pd/C,
\ ~ EtOH
O N~'~-OH 2) BrCN, MeOH,
NO2 H
14-2 71 /a for 2 steps
O 0
~10 1) SOCI2880 C
O
2) K2CO3, TBAI (cat.), \ ~ \ I
O ~
N-\,'OH DMF, 100 C O N~
N~NH2 N'~Nl
14-3 14-4 H
33% for 2 steps
0
NaOH, MeOH, 60 C HO p-alanine t-Bu ester*HCI, HOBt,
\ ! O \ ! N EDC,DMF
N==~, N 32% for 2 steps
14-5 H
0 0 0 0
XO" ~N 50% TFA/DCM HO" v 'N
H ) H ~
J:: N~ ~ O J:: -
O
N~N.~
14-6 H *TFA H
14
Step 1:
[001371 Following the procedure described for step 3 in example 8, 4-hydroxy-
3,5-dimethylbenzoic acid methyl ester (708 mg; 3.93 mmol) was reacted with 2,6-
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difluoronitrobenzene giving, after purification by FCC (silica gel; elution
with 1:1
hexanes:EtOac), 1.3 g (100%) of 14-1.
StM2.
[00138] To a solution of 14-1 (320 mg; 1.00 mmol) in MeOH (4 mL) was added
ethanolamine (0.6 mL; 10 mmol) and the reaction mixture was heated to 80 C in
a
sealed pressure vessel for 16 h. The mixture was then cooled and concentrated
in
vacuo and the crude residue was purified by FCC (silica gel; elution with 1:1
hexanes:EtOAc) to afford 228 mg (63%) of 14-2.
Data for 14-2: MS: m/z (assignment, relative intensity) 361.1 (1VI+H+, 100).
StM 3:
[00139] Following the procedure described for step 5 in example 8, compound
14-2 from above was reduced and then reacted with cyanogen bromide to afford,
after purification by FCC (silica gel; elution with 10% MeOH/DCM), 159 mg (71%
for 2 steps) of aminobenzimidazole 14-3.
Data for 14-3: MS: m/z (assignment, relative intensity) 356.2 (M+H+, 100).
Step 4:
[00140] 14-3 (52 mg; 0.146 mmol) was treated with thionyl chloride (1.5 mL)
and the resultant mixture was heated to 80 C for 0.5 h. The mixture was then
cooled and concentrated in vacuo and the residue was dissolved in DMF (1.5
mL).
To this was added K2C03 (101 mg; 0.73 mmol) and a crystal of
tetrabutylammonium iodide and the reaction mixture was heated to 100 C for 2
d.
The mixture was then cooled and partitioned between EtOAc and water. The
aqueous phase was extracted with EtOAc (2X) and the organic phases were
combined and washed with water (IX) and brine (1X). This was dried (Na2SO4),
filtered, and concentrated in vacuo and the crude residue was purified by FCC
(silica gel; elution with 5% MeOH/EtOAc) giving 16 mg (33% for 2 steps) of 14-
4.
Data for 14-4: MS: na/z (assignment, relative intensity) 338.3 (M+H', 100).
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Step 5:
[00141] 14-4 (16 mg; 0.0474 mmol) in MeOH (1 mL) was treated with 1 N
NaOH (aq) (142 L; 0.142 mmol) and the reaction mixture heated to 60 C for 4
h.
The mixture was cooled, concentrate in vacuo and treated with 1 N HCl (aq)
(142
L) forming a thick white precipitate which was collected by filtration. The
crude
acid 14-5 was used in the next step without further purification.
Data for 14-5: MS: m/z (assignment, relative intensity) 324.4 (M+H+, 100).
Step 6:
[00142] Following the procedure described for step 1 in example 3, 14-5 was
coupled with 0-alanine t-Bu ester hydrochloride giving, after purification by
FCC
(silica gel; elution with 10% MeOH/DCM), 7 mg (32%) of 14-6.
Data for 14-6: MS: m/z (assignment, relative intensity) 451.1 (M+W, 83), 395.2
(M-tBu, 100), 923.1 (2M+Na , 17).
Step 7:
[00143] 14-6 (4mg; 0.0094 mmol) was treated with 50% TFA/DCM and allowed
to stand for 16 h. This was then concentrated in vacuo and triturated with
Et20
giving 6.6 mg (84%) of 14 as a TFA salt.
Data for 14: MS: m/z (assignment, relative intensity) 394.44 (M+H+, 100).
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Example 15:
cl 0
(3-alanine ethyl ester*HCI, K2C03, fj ~
Os TBAB (cat), DMF, 80 oC O \ I O~
O O 33%
15-1
0
BBr3, DCM, 2,6-difluoronitrobenzene,
-78 C to 23 C; CsCO3, DMF, 80 C
EtOH 0 OH
15-2 100% for 2 steps
0
methylamine, DMF, 23 C
N \( qF
O0 NO2 38%
15-3
0 1) H2 (g; 1 atm), 10% Pd/C,
~O~ v N / I / I EtOH
O~O ~ N 2) BrCN, MeOH
NO2 H
15-4 21 % for 2 steps
0
~OJ~N s / IN NaOH (aq), MeOH, 60 C;
O \ N~ H +
N -~
15-5 NH2 100%
0
O%0
N~
NH2 *TFA
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Step 1:
[00144] To a solution of (2-Chloromethyl-5-methoxy-phenyl)-acetic acid ethyl
ester (248 mg; 1.02 mmol) in DMF (1 mL) was added (3-alanine ethyl ester
hydrochloride (188 mg; 1.23 mmol), K2C03 (423 mg; 3.06 mmol), and
tetrabutylammonium bromide (small scoop). The reaction mixture was heated to
70
C for 1 h and then the temperature was.increased to 80 C and stirring was
continued for a further 24 h. The mixture was cooled, diluted with EtOAc and
washed with 1N HCl (aq) (1X), water (1X) and brine (1X). The organic phase was
dried (Na2SO4), filtered and concentrated in vacuo. The crude residue was
purified
by FCC (silica gel; elution with 2:1 EtOAc:hexanes) giving 92 mg (33%) of 15-
1.
Step 2:
[00145] Following the general procedure described for step 2 in example 8, the
product from above, 15-1 (50 mg; 0.180 mmol), was reacted with BBr3 giving
phenol 15-2 which was used in the next step crude without fnrther
purification.
Step 3:
[00146] Following the procedure described for step 3 in example 8, the crude
product from above, 15-2, was reacted with 2,6-difluoronitrobenzene giving,
after
purification by FCC (silica gel; elution with 1:2 hexanes:EtOac), 72 mg (100%
for
2 steps) of 15-3.
Data for 15-3: MS: m/z (assignment, relative intensity) 403.2 (M+H+, 80),
425.2
(M-tBu, 45).
Stei~ 4:
[00147] Following the procedure described for step 4 in example 8, compound
15-3 from above was reacted with methylamine to afford 28 mg (38%) of 15-4
after
purification by FCC (silica gel; elution with 1:2 hexanes:EtOAc).
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Step 5:
[00148] Following the procedure described for step 5 in example 8, compound
15-4 from above was reduced and then reacted with cyanogen bromide to afford,
after purification by FCC (silica gel; elution with 10% MeOH/DCM), 6 mg (21 %
for 2 steps) of aminobenzimidazole 15-5.
Data for 15-5: MS: fn/z (assignment, relative intensity) 409.2 (M+H+, 100).
Step 6:
[00149] To a solution of ethyl ester 15-5 (6 mg; 0.0147 mmol) in MeOH (1 mL)
was added 1 N NaOH (aq) (44 L; 0.044 mmol). The resultant mixture was heated
to 60 C for 3h. The mixture was then cooled and concentrated in vacuo and the
crude material was acidified to pH 2 with 1 N HCl (aq). The mixture was
applied
to a column filled with Dowex 50 W(H) ion exchange resin and then eluted with
water until the eluent became neutral (pH 6). The column was then eluted with
5%
pyridine/HzO. Fractions containing the desired product were pooled and
concentrated in vacuo and the zwitterion thus obtained was treated with 50%
TFA/DC1V1 and concentrated in vacuo giving 7.5 mg of 15 (100%) as the TFA
salt.
Data for 15: MS: m/z (assignment, relative intensity) 381.3 (M+H+, 100).
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Exam 1be16:
I
O 0 3-amino-9-propanol,
N a EtOH, 80 C o ~ ~
NO2 66%
13-5
O O 1) H2 (g; I atm), 10% Pd/C,
EtOH
N
2) BrCN, MeOH
NO2 H OH 62% for 2 steps
16-1
1 ~
O ~ 0 1) SOC12, 80 C
~,O N )aj: 2) K?CO3, TBA! (cat.)
a DMSO, 120 C
~OH
16-2 NH2 30% for 2 steps
O 0 IN NaOH (aq), MeOH, 60 C;
ao N H+
N==( 45%
16-3 HND
Co
o HO N a
O N
N=-{
16 HN
*HCI
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WO 2007/001249 PCT/US2005/020656
~ \
0 ~ 0 3-amino-l-propanol,
-,-~-O N EtOH, 80 C
\ O ~ F
NO2 66%
13-5
O O 1) Hz (g; 1 atm), 10% Pd/C,
EtOH
N aO"1:: 2) BrCN, MeOH
H~
NO2 OH 62% for 2 steps
16-1
~ \
/
O O 1) SOCI2, 80 C
O N /
~ 2) K2C03, TBAI (cat.)
~ O DMSO, 120 C
N~ ~OH
16-2 NH2 30% for 2 steps
~ \
O ~ 0 IN NaOH (aq), MeOH, 60 C;
N a O N H+
N~/ 45%
~
16-3 H\N
O O
HO N a
O N~
N~
HN
*HCI
16
Step 1:
[00150] To a solution 13-5 [For preparation see example 13 above] (341 mg;
0.713 mmol) in anhydrous EtOH (5 mL) was added 3-amino-l-propanol (0.55 mL;
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7.13 mmol). The reaction mixture was heated to 80 C for 16 h. The mixture was
then cooled and concentrated in vacuo. The crude residue was purified by FCC
(silica gel; elution with 2:1 EtOAc:hexanes) giving 250 mg of 16-1 (66%)
Step 2:
[00151] Following the general procedure described for step 5 in example 8,
compound 16-1 from above was reduced and then reacted with cyanogen bromide
to afford, after purification by FCC (silica gel; elution with 5% MeOH/DCM
then
10% MeOH/DCM), 144 mg (62% for 2 steps) of aminobenzimidazole 16-2.
Data for 16-2: MS: nz/z (assignment, relative intensity) 529.2 (M+H+, 100),
1057.0
(2M+H+, 15).
Sten3:
[00152] 16-2 (100 mg; 0.19 mmol) was treated with thionyl chloride (2 mL) and
the resultant mixture was heated to 80 C for 1 h. The mixture was then cooled
and
concentrated in vacuo and the residue was dissolved in DMSO (4 mL). To this
was
added K2C03 (131 mg; 0.95 mmol) and a small scoop of tetrabutylammonium
iodide and the reaction mixture was heated to 135-140 C for 5 h. The heat was
then reduced to 120 C and stirring was continued for 16 h at this
temperature. The
mixture was then cooled, diluted with EtOAc (50 mL) and washed with water (2 X
mL) and brine (1X 10 mL). The organic phase was dried (Na2SO4), filtered, and
concentrated in vacuo. The crude residue was purified by FCC (silica gel;
elution
with 2% MeOH/DCM then 10% MeOH/DCM) giving 30 mg (30% for 2 steps) of
16-3 along with 27 mg of recovered 16-2 (resulting from hydrolysis of
intermediate
chloride).
Data for 16-3: MS: m/z (assignment, relative intensity) 511.2 (M+H+, 100),
1020.9
(2M+H+, 25).
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St~ 4:
[00153] To a solution of ethyl ester 16-3 (30 mg; 0.0588 mmol) in MeOH (1
mL) was added 1 N NaOH (aq) (0.2 mL; 0.2 mmol). The resultant mixture was
heated to 60 C for 3h. The mixture was then cooled and concentrated in vacuo
and
the crude material was acidified to pH 2 with 1 N HCl (aq). The mixture was
applied to a column filled with Dowex 50 W(FI) ion exchange resin and then
eluted with water until the eluent became neutral (pH 6). The column was then
eluted with 5% pyridine/H20. Fractions containing the desired product were
pooled
and concentrated in vacuo and the zwitterion thus obtained was suspended in
Et20
and HCl gas bubbled through to make the HCl salt. This was concentrated in
vacuo
and dried in a vacuum oven at 60 C for overnight giving 14 mg of 16 (45%) as
the
HCl salt.
Data for 16: MS: nz/z (assignment, relative intensity) 483.2 (M+H+, 100),
965.0
(2M+IH+, 8).
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Example 17:
0-\
O
O o
q
H
N piperonal, benzotriazole, N-N 6aoTsOH (cat), toluene, reflux, '_~Zz
1 , -H20 (Dean-Stark) ~ ,
11-2 55%
17-1
O--\
TBDMSO
>__ _O Yb(OTf)2, DCM 0
ij0
~O N ao
75% 17-2
O-\
O
1) H2 (1 atm), 10% Pd/C, MeOH
0 0
2) 2,6-difluoronitrobenzene, Cs2CO3, __O N / ~
DMF, 80 C \~ O\ I F
67% for 2 steps NO2
17-3
O-\
O
3-amino-1-propanol,
O O
EtOH, 80 C ~O N
64% \ I O \ ( N
NO2 H~-
17-4 OH
0--\
0
1) H2 (g; 1 atm), 10% Pd/C, I\
EtOH O ~ O
2) BrCN, MeOH \O N a
11
62% for 2 steps O '\-\
N~ OH
17-5 NH2
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O-\
0
OO 1) SOC12, 80 C
O N 2) DIPEA, NMP, 120 C
O N
N=~ ~OH 57% for 2 steps
17-5 NH2
O-'\
0
\
O' ~ O IN NaOH (aq), MeOH, 60 C;
O N H+
a
0 N 69%
17-6 N HN
D
O-\
0
O O
HO N \ ( \ I
O N
N=~
HN
*HCI
17
St~1:
[00154] A mixture of 11-2 [For preparation see example 11 above] (253 mg;
1.00 mmol), piperonal (150 mg; 1 mmol), benzotriazole (119 mg; 1 mmol) and
TsOH*H20 (19 mg; 0.1 mmol) was heated to reflux in 5 mL of toluene with a
Dean-Stark trap for 16 h. The rnixture was cooled, diluted with DCM (100 mL)
and washed with I N NaOH (aq) (1 X 50 mL). The aqueous phase was back-
extracted with DCM (2 X 50 mL) and the combined organic phases were dried
CA 02611971 2007-12-12
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(Na2SO4), filtered and concentrated in vacuo. The crude residue was purified
by
FCC (silica gel; elution with 2:1 hexanes:EtOAc) to afford 274 mg (55%) of 17-
1.
Ste,p 2:
[00155] To a solution of 17-1 (274 mg; 0.544 mmol) in DCM (5 mL) were added
ytterbium trifluoromethanesulfonate [Yb(OTf)3] (67 mg; 0.109 mmol) and 1-(tert-
butyldimethylsilyloxy)-1-methoxyethene (0.15 mL; 0.815 mmol). The resultant
mixture was stirred at room temperature for 16 h. The mixture was then
transferred
to a separatory fimnel and washed with water (1X). The aqueous phase was back-
extracted with DCM (2X) and the combined organic phases were dried (Na2SO4),
filtered and concentrated in vacuo. The crude residue was purified by FCC
(silica
gel; elution with 2:1 hexanes:EtOAc) giving 187 mg (75%) of 17-2.
Step 3:
100156] 17-2 (187 mg; 0.407 miriol) was debenzylated by hydrogenation in
MeOH in the presence of a catalytic amount of 10% Pd/C (1 small scoop) under I
atm of H2 (g) (balloon) for 3 d. The reaction mixture was filtered through a
pad of
Celite aaid concentrated in vacuo. The crude phenol was taken up in DMF (4 mL)
and 2,6 difluoronitrobenzene (43 L; 0.407 mmol) and Cs2CO3 (133 mg; 0.407
nnmol) were added. The mixture was heated to 80 C for 16 h. After cooling to
room temperature the mixture was filtered through a pad of Celite,
concentrated in
vacuo and the crude residue was purified by FCC (silica gel; elution with 3:1
hexanes:EtOAc) to afford 139 mg (67% for 2 steps) of 17-3.
Step4:
[00157] To a solution 17-3 (139 mg; 0.273 mmol) in anhydrous MeOH (3 mL)
was added 3-amino-1 -propanol (0.21 mL; 2.73 mmol). The reaction mixture was
heated to 80 C for 16 h. The mixture was then cooled, concentrated in vacuo
and
partitioned between EtOAc and water. The aqueous phase was extracted with
EtOAc (1X) and the combined organic phases were then washed with water (IX)
and brine (1X). The organic phase was then dried (Na2SO4), filtered and
concentrated in vacuo. The crude residue was purified by FCC (silica gel;
elution
with 3:1 EtOAc:hexanes) giving 98 mg (65%) of 17-4.
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Step 5:
[00158] Following the general procedure described for step 5 in example 8,
compound 17-4 from above was reduced and then reacted with cyanogen bromide
to afford, after purification by FCC (silica gel; elution with 5% MeOH/DCM
then
10% MeOH/DCM), 63 mg (65% for 2 steps) of aminobenzimidazole 17-5.
Data for 17-5: MS: nz/z (assignment, relative intensity) 559.2 (M+IT', 100),
1116.9
(2M+H'-, 36).
Step 6:
[00159] 17-5 (63 mg; 0.113 mmol) was treated with thionyl chloride (3 mL) and
the resultant mixture was heated to 80 C for 45 min. The mixture was then
cooled
and concentrated in vacuo and the residue was dissolved in NMP (2.5 mL). To
this
was added a scoop of 4 A molecular sieves and DIPEA (98 L; 0.563 mmol) and
the reaction mixture was heated to 120 C for 6.5 h. More DIPEA (98 L; 0.563
mmol) was then added and the mixture was heated at 120 C for an additional 9
h.
The mixture was then cooled and partitioned between EtOAc and water. The
aqueous phase was extracted with EtOAc and the organic phases were combined
and washed with water (1X) and brine (1X). The organic phase was dried
(Na2SO4), filtered, and concentrated in vacuo. The crude residue was purified
by
FCC (silica gel; elution with 2% MeOH/DCM then 5% MeOH/DCM) giving 35 mg
(57% for 2 steps) of 17-6.
Data for 17-6: MS: fn/z (assignment, relative intensity) 541.2 (M+H, 100).
Step 7:
[00160] To a solution of methyl ester 17-6 (35 mg; 0.0647 mmol) in MeOH (2
mL) was added 1 N NaOH (aq) (0.32 mL; 0.32 mmol). The resultant mixture was
heated to 60 C for 1.5 h. The mixture was then cooled to room temperature and
stirring continued for 16 h. The mixture was then warmed back to 60 C for an
additional 1 h, cooled and concentrated in vacuo. The crude material was
acidified
to pH 5 with 1 N HCl (aq) (0.32 mL; 0.32 mmol) forming a white precipitate
which
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was collected by filtration and washed with cold water. The material thus
obtained
was treated with 2 N HCl in EtaO, concentrated in vacuo , and then dried in a
vacuum oven at 70 C for 3 h to give 25 mg of 17 (69%) as the HC1 salt.
Data for 17: MS: ni/z (assignment, relative intensity) 527.1 (M+H+, 100),
1052.8
(2M+H'-, 45).
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Example 18:
II 0 0
HN
O NaH, DMF
112 100% 18-1 ~
I ~ O O
H2, Pd/C 0 O F / F
EtOH a N \ /
N02 I/ O\( F
90% OH K2CO3i DMF NO2
18-2 74% 18-3
0 0
H2N-'- ~OH '
DIEA, EtOH, d O\ H~'OH
94% 18-4 NO2
0 0
Zn, AcOH 0~~N CNBr
THF 0 EtOH
NH2 H .49%
18-5
O 0
/--,0,~~N \ 1) SaC12
I~ O\ I N~ 2) DIEA, DME, a
N OH 5%
18-6 NH2
O 0 0 0
6N HCI HO" ~N ao-P(N-
N quant. _ J N-
18-7 HN
18 *HCI
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step 1:
[00161] To 11-2 [For preparation see example 11 above] (0.25g, 1 mmol) in
l Oml dry DMF was added NaH (0.08g, 2mmol). After 2.5 hours
ethylbromopropionate (0.36g, 2mmol)in 5ml dry DMF was added dropwise. After
2 hours the reaction was quenched with 0.1N HCl and concentrated in vacuo. The
residue was purified on a silica gel column, eluting with 5 foMeOH/DCM to
yield
18-1 (0.36g, 1 mmol, 100%) as an oil.
Step 2:
[00162] To 18-1 in 5ml EtOH was added a small scoop of 10% Pd/C and placed
under a H2 balloon. The mixture was stirred at room temperature for 18 hours.
The
mixture was then filtered and concentrated in vacuo to yield 18-2 (0.24g, 0.9
mmol,
90%) as an oil.
Step3:
[00163] A mixture of 18-2 (0.24g, 0.9mmo1), 2,6-difluoronitrobenzene (0.24g,
1.5mmol), and K2C03 (0.28g, 2mmol) was stirred at 80 C in lOml DMF for 18
hours. The mixture was then filtered and concentrated in vacuo to yield 18-3
(0.27g, 0.67mmol, 74%) as an oil which was used without further purification.
Step 4:
[00164] A solution of 18-3 (0.27g, 0.67mmol), 3-amino-l-propapol (0.51m1,
6.7mmol) and DIEA (0.57m1, 3.3mmol) was heated to reflux in DME for 2 hours.
The solution was then concentrated in vacuo and the residue purified on a
silica gel
column, eluting with 5% MeOH/DCM to yield 18-4 (0.29g, 0.63mmo1, 94%) as an
oil.
Step 5:
[00165] To 18-4 (0.29g, 0.64mmol) in 15m1 TH was added 2ml acetic acid and
0.5g of zinc dust. The mixture was heated at reflux for 15 minutes. The
mixture
was cooled to room temperature, filtered and concentrated. He residue was
taken
up in DCM and washed with 1N NaOH and brine, dried over MgSO4 and
concentrated to yield 18-5 which was used without further purification.
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Ste6:
[00166] To crude 18-5 in 10m1 EtOH was added cyanogen bromide (0. l Og,
lmmol) and stirred at room temperature for 18 hours. The solution was then
concentrated and the residue purified on a silica gel column, eluting with 7%
MeOH/DCM to yield 18-6 (0.14g, 0.31mmol, 49%) as an oil.
St__l? 7
[00167] To 18-6 was added lOml SOC12 and stirred at room temperature for 30
minutes. The solution was concentrated, taken up in 15ml DME and basified with
DIEA. The solution was then heated at reflux for 48 hours. The solution was
then
concentrated and the residue purified on a silica gel column, eluting with 10%
MeOHIDCM. The resulting oil was then purified further by preparative HPLC to
yield 18-7 (0.007g, 0.016mmo1, 5%) as an oil.
Step 8:
[00168J 18-7 (4mg, 0.009mo1) was treated with 4m16N HCl for 18 hours. The
mixture was then concentrated in vacuo to yield 18 (4mg, 0.009mmol, 100%) as a
white solid.
Data for 18: MS: rn/z (assignment, relative intensity) 407.2 (M+W, 95)
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Example 19:
0
o
0 0
HO H *HCI O,~N
,~ O '
O N' N HOBt, EDC, Et3N, r.t. N' N
~
~NH2 NH2
12%
2"5 19-1
O
6N HCI (aq), 23 C, 16 h O} N ,~
H0 O
N-
100% N~
NH2 *HCI
19,
steb 1:
[00169] To a solution of 2-5 (87 mg; 0.28 mmol) in DMF (1 mL) were added
triethylamine (194 L; 1.4 mmol), 1-hydroxybenzotriazole (56.7 mg; 0.42 mmol),
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (80.5 mg; 0.42
mmol) and pyrrolidine-3-carboxylic acid ethyl ester (50 mg; 0.35 mmol). The
reaction mixture was stirred at room temperature overnight. Then the mixture
was
partitioned between EtOAc and water. The aqueous phase was extracted with
EtOAc (3X). The organic phases were combined and concentrated in vacuo. The
residue was purified by FCC (silica gel; elution with 10% MeOH/DCM) giving 15
mg (12%) of 19-1.
Data for 19-1: MS: m/z (assigmnent, relative intensity) 437.2 (M+H+, 100).
Stm 2:
[00170] 19-1 (15 mg; .034 mmol) was treated with 1 mL of 6 N HCl (aq) and
stirred at room temperature for 16 h. This was then concentrated in vacuo and
dried
in a vacuum oven at 60 C for 16 h giving 15 mg (100%) of 19.
Data for PS580985: MS: tta/z (assignment, relative intensity) 409.2 (M+H+,
100).
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[00171] A good review of the demonstrated utility in vivo of vitronectin
antagonists is provided by Miller et al. [Drug Discovery Today, 5, 397-408
(2000)].
The article is incorporated herein by reference. The article describes a
number of
compounds that have been tested in. vivo for the treatment of osteoporosis,
tumors,
rheumatoid arthritis, and restenosis based on their vitronectin inhibitory
activity in
vitro. Therefore compounds showing the vitronectin inhibitory activity in
vitro
described below would be expected by persons of skill to exhibit utility for
these
conditions in vivo.
The compounds were tested according to the following procedure:
[00172] Assays of human placental avP3 and av(35 were performed as described
in the literature [Pytela et al. "Methods In Enzymology, 144, 475-489 (1987)]
with
minor modification. Time-resolved fluorescence was used as an alternative to
colorimetric detection to improve the assay sensitivity and the signal to
noise ratio.
In the assay, 75 ng of either av(33 or av(35 was coated on the microtiter
wells.
Various concentrations of the test compound were added, together with 5 nmol
of
europium-labeled vitronectin. The samples were allowed to incubate at room
temperature for one hour. Following incubation, the wells were emptied and
washed. The amount of europium- labeled vitronectin bound to the plate was
determined by time-resolved fluorescence measurement. A 384 well plate was
coated by incubating each well with 25 L of a 3 g per mm solution of av(33
(75 ng
per well) in buffer A. Buffer A is an aqueous solution containing 50 mmol tris-
hydrochloride, 100 mm.ol sodium chloride, 1 mmol magnesium chloride, 1 mmol
magnesium chloride, 1 mmol calcium chloride, I mmol manganese chloride
adjusted to pH7.4. The plate was incubated overnight at 4 C. 70 L of 1%
bovine
serum albumin (BSA) per well was added and the plate incubated for three hours
at
room temperature, followed by washing three times with 70 L of Buffer A per
well. To each well was added 15 L of a solution of test compound in 46 L of
buffer A containing 1% dimethylsulfoxide, 20 mmol
diethylenetriaminepentaacetic
acid (DTPA) and 0.05% BSA (1-5 M final concentration of test compound). Also
added was 5 L of 20 nm Eu3+ vitronectin in Buffer A with 20 mmol DTPA and
0.05% BSA (to a 5 nm final concentration). The plate was incubated for one
hour
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at room temperature and washed three times with 70 L each of Buffer A. Twenty-
five L of a standard fluorescence enhancement solution was added and the
fluorescence measured and compared to control. The IC50 of a test compound
represents the concentration of that compound required to suppress
fluorescence by
50%.
[001731 All of the compounds of the preceding examples and in the following
table, which were made by analogous procedures, exhibited IC50's below 1 M
against av(33 integrin and below 10 M against av(3s integrin.
Example Structure
number
20 NHZ
N-~
O N---
I \
HO N /
~~
O O
21
O
O N ao HO \ N~
N-=-~
N H2
22 a O
H
O \ I
N~
NH2
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F O
123 S..",
HO I \ / ~
O \ N f
N-=-
NH2
24 0 0
oJ,-~ N O
H \ ~ \ I
O N'
NH2
25 O O
H \ ~ \ f
O N.-
NH2
26 O 0
H
O
N~
NH2
27 0
HO
0 N--
O N-=--~
NH2
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28
NH2
\O N--~
H N--,_,
~ \ I \
O
'~!1
O O
29 0
HO O
O N...--
N~(
NH2
0 ~
I ~,,,
0
HO ~
0
N.,,
NH2
31
NH2
C! N_
o N-_..,.
l ~
HO ~
N
O 0
32 0 0
HO''~N
H
O
N
HN
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33 0
HO CH3 /' ~ O N-"CH3
CH3 N=,
NH2
34 O
Ho o f o
O N,-
O
NH2
35 0 0
HA~N OCH3
H
0 s
OCH3 N=~
NH2
36 0 0 F
HO" V ' N OCH3
H
0 N-CH3
OCH3 N'~
NH2
37 0
0 OH
N,.GH
H
O
N
H2N-~/
N
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38 O O
HO' v 'N
H
O N--
NH2
39 0
0 OH
O y~CH
O
N :]6
OH2No N
40 O O
H \ , \ I
O N~
N={
NH2
41 0 0
HO' v 'N
H O NH
N=~
NH2
42 0
HO O
N
ao N--
N==Z(
NH2
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43 o 0
H F3
Hp'~I~N =~ C
/, p
N~
NH2
44 ~ N
(
O O
HO N ..---
N
NH2
45 0 0
\ OMe
HO H
/ p -CH
N 3
OMe N:-=<
NHCH3
46 NH2
O 0 N_~
N-.
HO H O
47 p p
HO'~N
H
/ p \ N,-
O N:::~<
NH2
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48 O 0
HO' v 'N
O N/
N~
NH2
49 ~ F
~ /
O O
O
HO N
lxxl O N
NH2
50 0 0
HO' v 'N
H \I \I
O N
/\
N- CF3
NH2
51 NH2
N O/
D6,
O
O
0
HN
I-Y O
OH
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52 O-\
0
( \
O ~ O
HO H / /
O
N
NH2
53
O O
HO)L",~ N
O NH H I / ~ ( ---
~ O N
O N==~
NH2
91
, , ,
