Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
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INHIBITORS OF FACTOR Xa
Related Applications
This application claims benefit of priority under 35 USC ~ 119(e) to U.S.
Provisional Application No. 60/13~,~i9 filed on May 24, 1999, which is herein
incorporated in its entirety by reference.
Field of the Invention
This invention relates to novel compounds which are potent and highly
selective inhibitors of isolated factor Xa or when assembled in the
prothrombinase
complex. These compounds show selectivity for factor Xa versus other proteases
of
the coagulation (e.g. thrombin, fVIIa, flXa) or the fibrinolytic cascades
(e.g.
plasminogen activators, plasmin). In another aspect, the present invention
relates to
novel monoamidino-containing compounds, their pharmaceutically acceptable
salts,
and pharmaceutically acceptable compositions thereof which are useful as
potent
and specific inhibitors of blood coagulation in mammals. In yet another
aspect, the
invention relates to methods for using these inhibitors as therapeutic agents
for
disease states in mammals characterized by coagulation disorders.
Background of the Invention
Hemostasis, the control of bleeding, occurs by surgical means, or by the
physiological properties of vasoconstriction and coagulation. This invention
is
particularly concerned with blood coagulation and ways in which it assists in
maintaining the integrity of mammalian circulation after injury, inflammation,
disease, congenital defect, dysfunction or other disruption. Although
platelets and
blood coagulation are both involved in thrombus formation, certain components
of
the coagulation cascade are primarily responsible for the amplification or
acceleration of the processes involved in platelet aggregation and fibrin
deposition.
Thrombin is a key enzyme in the coagulation cascade as well as in
hemostasis. Thrombin plays a central role in thrombosis through its ability to
catalyze the conversion of fibrinogen into fibrin and through its potent
platelet
activation activity. Direct or indirect inhibition of thrombin activity has
been the
focus of a variety of recent anticoagulant strategies as reviewed by Claeson,
G.,
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"Synthetic Peptides and Peptidomimetics as Substrates and Inhibitors of
Thrombin
and Other Proteases in the Blood Coagulation System", Blood Coag. Fibrinol.
5_,
411-436 (1994). Several classes of anticoagulants currently used in the clinic
directly or indirectly affect thrombin (i.e. heparins, low-molecular weight
heparins,
heparin-like compounds and coumarins).
A prothrombinase complex, including Factor Xa (a serine protease, the
activated form of its Factor X precursor and a member of the calcium ion
binding,
gamma carboxyglutamyl (Gla)-containing, vitamin K dependent, blood coagulation
glycoprotein family), converts the zymogen prothrombin into the active
procoagulant thrombin. Unlike thrombin, which acts on a variety of protein
substrates as well as at a specific receptor, factor Xa appears to have a
single
physiologic substrate, namely prothrombin. Since one molecule of factor Xa may
be
able to generate up to 138 molecules of thrombin (Elodi et al., Thromb. Res.
15, 617-
619 (1979)), direct inhibition of factor Xa as a way of indirectly inhibiting
the
formation of thrombin may be an efficient anticoagulant strategy. Therefore,
it has
been suggested that compounds which selectively inhibit factor Xa may be
useful as
in vitro diagnostic agents, or for therapeutic administration in certain
thrombotic
disorders, see e.g., WO 94/13693.
Polypeptides derived from hematophagous organisms have been reported
which are highly potent and specific inhibitors of factor Xa. United States
Patent
4,588,587 describes anticoagulant activity in the saliva of the Mexican leech,
Haementeria o~cinalis. A principal component of this saliva was shown to be
the
polypeptide factor Xa inhibitor, antistasin (ATS), by Nutt, E. et al., "The
Amino
Acid Sequence of Antistasin, a Potent Inhibitor of Factor Xa Reveals a
Repeated
Internal Structure", J. Biol. Chem., 2_6~, 10162-10167 (1988). Another potent
and
highly specific inhibitor of Factor Xa, called tick anticoagulant peptide
(TAP), has
been isolated from the whole body extract of the soft tick Ornithidoros
moubata, as
reported by Waxman, L., et al., "Tick Anticoagulant Peptide (TAP) is a Novel
Inhibitor of Blood Coagulation Factor Xa" Science, 248, 593-596 (1990).
Factor Xa inhibitory compounds which are not large polypeptide-type
inhibitors have also been reported including: Tidwell, R.R. et al.,
"Strategies for
Anticoagulation With Synthetic Protease Inhibitors. Xa Inhibitors Versus
Thrombin
Inhibitors", Thromb. Res., 19, 339-349 (1980); Turner, A.D. et al., "p-Amidino
Esters as Irreversible Inhibitors of Factor IXa and Xa and Thrombin",
Biochemistry,
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25, 4929-4935 (1986); Hitomi, Y. et al., "Inhibitory Effect of New Synthetic
Protease Inhibitor (FUT-175) on the Coagulation System", Haemostasis, 1 S, 164-
168 (1985); Sturzebecher, J. et al., "Synthetic Inhibitors of Bovine Factor Xa
and
Thrombin. Comparison of Their Anticoagulant Efficiency", Thromb. Res., 54, 245-
252 (1989); Kam, C.M. et al., "Mechanism Based Isocoumarin Inhibitors for
Trypsin and Blood Coagulation Serine Proteases: New Anticoagulants",
Biochemistry, ~, 2547-2557 (1988); Hauptmann, J. et al., "Comparison of the
Anticoagulant and Antithrombotic Effects of Synthetic Thrombin and Factor Xa
Inhibitors", Thromb. Haernost., 63, 220-223 (1990); and the like.
Others have reported Factor Xa inhibitors which are small molecule organic
compounds, such as nitrogen containing heterocyclic compounds which have
amidino substituent groups, wherein two functional groups of the compounds can
bind to Factor Xa at two of its active sites. For example, WO 98/28269
describes
pyrazole compounds having a terminal C(=NH)-NHZ group; WO 97/21437 describes
benzimidazole compounds substituted by a basic radical which are connected to
a
naththyl group via a straight or branched chain alkylene,-C(=O) or -S(=O)Z
bridging
group; WO 99/10316 describes compounds having a 4-phenyl-N-alkylamidino-
piperidine and 4-phenoxy-N-alkylamidino-piperidine group connected to a 3-
amidinophenyl group via a carboxamidealkyleneamino bridge; and EP 798295
describes compounds having a 4-phenoxy-N-alkylamidino-piperidine group
connected to an amidinonaphthyl group via a substituted or unsubstituted
sulfonamide or carboxamide bridging group.
There exists a need for effective therapeutic agents for the regulation of
hemostasis, and for the prevention and treatment of thrombus formation and
other
pathological processes in the vasculature induced by thrombin such as
restenosis and
inflammation. In particular, there continues to be a need for compounds which
selectively inhibit factor Xa or its precursors. Compounds that have different
combinations of bridging groups and functional groups than compounds
previously
discovered are needed, particularly compounds which selectively or
preferentially
bind to Factor Xa. Compounds with a higher degree of binding to Factor Xa than
to
thrombin are desired, especially those compounds having good bioavailability
and/or
solubility.
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Summary of the Invention
The present invention relates to novel compounds which inhibit factor Xa,
their pharmaceutically acceptable isomers, salts, hydrates, solvates and
prodrug
derivatives, and pharmaceutically acceptable compositions thereof which have
particular biological properties and are useful as potent and specific
inhibitors of
blood coagulation in mammals. In another aspect, the invention relates to
methods
of using these inhibitors as diagnostic reagents or as therapeutic agents for
disease
states in mammals which have coagulation disorders, such as in the treatment
or
prevention of any thrombotically mediated acute coronary or cerebrovascular
syndrome, any thrombotic syndrome occurnng in the venous system, any
coagulopathy, and any thrombotic complications associated with extracorporeal
circulation or instrumentation, and for the inhibition of coagulation in
biological
samples.
In certain embodiments, this invention relates to novel compounds which are
potent and highly selective inhibitors of isolated factor Xa when assembled in
the
prothrombinase complex. These compounds show selectivity for factor Xa versus
other proteases of the coagulation cascade (e.g. thrombin, etc.) or the
fibrinolytic
cascade, and are useful as diagnostic reagents as well as antithrombotic
agents.
In a preferred embodiment, the present invention provides a compound of the
formula I:
A-Y-D-E-G-J-Z-L
wherein:
A is selected from:
(a) phenyl, which is independently substituted with 0-2 R' substituents;
(b) a monocyclic or fused bicyclic heterocyclic ring system having from
5 to 10 ring atoms, wherein 1-4 ring atoms of the ring system are
selected from N, O and S, and wherein the ring system may be
substituted with 0-2 R' substituents;
(c) naphthyl, which is independently substituted with 0-2 R' substituents;
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(d) C,-C6-alkyl; C3-C8-cycloalkyl; and
(e) NRzR3, -C(=NRz)NRZR3, -NRZC(=NRz)NRZR3, -C(=NRz)R4, and
~zC(=~z)-Rs
R' is selected from:
Halo, C,~alkyl, Cz_6alkenyl, Cz_balkynyl, C3_8cycloalkyl, Co_4alkylC3_
8cycloalkyl,-CN, -NOz, -(CHz)mNRzR3, -C(=O)NRzR3, -C(-NRz)NRZR3,
-NRZC( NRz)NRZR3, -C(=NRz)R4 and NRZC(=NRz)-R3, -SOzNR2R3, -SOZRz,
-CF3, -ORz, and a S-6 membered aromatic heterocyclic system containing
from 1-4 heteroatoms selected from N, O and S, wherein from 1-4 hydrogen
atoms on the aromatic heterocyclic system may be independently replaced
with a member selected from the group consisting of halo, C,-C4 alkyl, -CN
C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_gcycloalkyl, Co_4alkylC3_gcycloalkyl
and
_NOz~
Rz and R3 are independently selected from the group consisting of
H, -OR'4, -NR'4R'S, C,~,alkyl, Cz_6alkenyl, Cz_balkynyl, C3_gcycloalkyl, Co_
4alkylC3_8cycloalkyl, COOC,~,alkyl, COO-Co_4alkylphenyl Co~alkylphenyl
and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on the ring atoms
of the phenyl and naphthyl moieties may be independently replaced with a
member selected from the group consisting of halo, C,~,alkyl, Cz_balkenyl, Cz_
6alkynyl, C3_gcycloalkyl, Co_4alky1C3_8cycloalkyl, -CN, and -NOz;
m is an integer of 0-2;
Y is a member selected from the group consisting of
a direct link, -C(=O)-, -N(R4)-, -C(=O)-N(R4)-, -N(R4)-C(=O)-, -SOz-, -O-,
-SOz-N(R4)-, N(R4)-SOz-, -C(=NR4), -C(=S)-, -CHz , -CHzN(R4)-;
R4 is selected from:
H, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_gcycloalkyl,
Co~alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~alkyl,
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Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~,alkylC3_gcycloalkyl, -CN, and
-NOz;.
D is a direct link or is a member selected from the group consisting of:
(a) phenyl, which is independently substituted with 0-2 R'a substituents;
(b) naphthyl, which is independently substituted with 0-2 R'a
substituents; and
(c) a monocyclic or fused bicyclic heterocyclic ring system having from
S to 10 ring atoms, wherein 1-4 ring atoms of the ring system are
selected from N, O and S, and wherein the ring system may be
substituted with 0-2 R'a substituents;
R'a is selected from:
Halo, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_$cycloalkyl, Co~alkylC3_
8cycloalkyl, -CN, -NOz, (CHz)",NRzaR3a, SOzNR2aR3a~ SOzRza~ CF3~ ORza, and
a 5-6 membered aromatic heterocyclic system containing from 1-4
heteroatoms selected from N, O and S, wherein from 1-4 hydrogen atoms on
the aromatic heterocyclic system may be independently replaced with a
member selected from the group consisting of halo, C,_4alkyl, Cz_balkenyl, Cz_
6alkynyl, C3_8cycloalkyl, Co~alkylC3_8cycloalkyl, -CN and -NOz.
m is an integer of 0-2;
Rza and R3a are independently selected from the group consisting of
H, C,~alkyl, Cz_balkenyl, Cz_6alkynyl, C3_$cycloalkyl, Co~alkylC3_8cycloalkyl,
Co~alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~alkyl,
Cz_6alkenyl, Cz_6alkynyl, C3_gcycloalkyl, Co_4a1ky1C3_8cycloalkyl, -CN and
_NOz~.
E is a member selected from the group consisting of:
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-N(RS)-COO)-~ -C(=O)-N(RS)-~ -N(RS)-C(=O)-N(R6)-~ -S02-N~5)-~
-N(RS)-SOZ-N(R6)- and N(RS)-SOZ-N(R6)-C(=O)-;
RS and R6 are independently selected from:
H, C,~,alkyl, CZ_6alkenyl, Cz_6alkynyl, C3_gcycloalkyl,
Co~alkylC3_8cycloalkyl,
Co~alkylphenyl, Co_4alkylnaphthyl, Co~alkylheteroaryl, C,~alkylCOOH and
C,~alkylCOOC,-alkyl, wherein from 1-4 hydrogen atoms on the ring atoms
of the phenyl, naphthyl and heteroaryl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~alkyl,
CZ_balkenyl, CZ_6alkynyl, C3_gcycloalkyl, Co~alkylC3_$cycloalkyl, -CN and
-NO2;
G is selected from:
-CR'R8- and -CR'aRBa-CbRBb-
Wherein R', R8, R'a, Rga, R'~ and Rgb are each independently a member selected
from
the group consisting o~
hydrogen, halo, -C,_6alkyl, haloalkyl, -CN, -NOz, -Cz_6alkenyl, -CZ_
6alkynyl, -C3_8cycloalkyl, -Co~,alkyl-C3_8-cycloalkyl, -Co~alkyl-CN, -Co_
4alkyl-NO2, -Co~alkyl-O-R9, -Co.~alkyl-S-R9, -Co~alkyl-S(=O)z-R9,
-Co~alkyl-S(O)-R9, -Co~alkyl-C(=O)-OR9, -Co~alkyl-C(=O)-N(R9a, R9b)
-Co~alkyl-C(=O)-R9, -Co.~alkyl-N(R9a, R9b), -Co-
4alkyl-N(-R9a)-C(=O)-R9b), -Co~alkyl-N(-R9a)-C(=O)-R9b, -Co-
4alkyl-N(-R9a)-C(=O)-N(-R9b), -Co~alkyl-N(-R9a)-S(=O)z-R9n, -Co_
4alkyl-S(=O)z-N(R9a, R9b), -Co~alkyl-S(=O)z-R9, -Co_
4alkyl-P(=O)(-OR9a)(-OR9b), -Co~,alkyl-N(-R9)-P(=O)(-OR9a)(-OR9b), -Co_
4alkyl-phenyl, -Co~alkyl-naphthyl, -Co~alkyl-heterocyclic ring system
containing from 1-4 heteroatoms selected from the group consisting of
O, N and S, wherein the heterocyclic ring system is a 5-6 membered
monocyclic ring or a 8-12 membered bicyclic ring, and wherein 0-4
hydrogen atoms of the phenyl ring, the naphthyl ring carbon and the
heterocyclic ring system are replaced by a member selected from the
group consisting of -C,_4alkyl, haloalkyl, halo, -CN, -NO2, -OR9~, -
SR9°,
-S(O)R9', -C(=O)-OR9', -C(=O)-N(-R9', R9a), -C(=O)-R9~, -N(R9°, R9d),
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-N(-R9o)-C(=O)-R9a~ -N(-R9~)-C(=O)-OR9a~ -N(-R9~)-C(=O)-N(-H~ R9a)
-N(-R9~)-SOZ-R9a, -SOZ-N(-R9', -R9a), -SOZ-R9c; or one of R', Rg, R'a, RBa,
R'~ and R8b can combine with a nitrogen on the E group to form a 5-7
membered heterocyclic ring containing a 0-3 additional heteroatoms
selected from the group consisting of O, N and S; or R'a and R'~ on
adjacent carbons combine to form a 3-6 membered carbocyclic ring;
R'~ and R8b combine to form alkylidene groups, such as HZC=, C,_
4a1ky1CH=, (C,~alkyl)ZC=, PhCH=;
R9, R9a, R9b' R9° and R9a are each independently a member selected from
the group
consisting of
H, halo -C,_balkyl, -CZ_6alkenyl, -Cz_6alkynyl, -C3_8cycloalkyl, -C~alkyl-C3_
gcycloalkyl, -CHZCHZOH, -CHzCH2-O-CH3, -Co~alkylphenyl, -Co_
4alkylheterocycle wherein the heterocycle may be a 5-6 membered ring, and
wherein from 0-4 hydrogen atoms from the ring atoms of the phenyl and
heterocycle groups may be independently replaced with a member selected
from the group consisting of halo, -C,~,alkyl, -CZ_6alkenyl, -Cz_6alkynyl,
-C3_8cycloalkyl, -Co~alkyl-C3_gcycloalkyl, -CN, -NO2, -C(=O)-OH, -C(=O)-O-
C,~alkyl, -C(=O)-NH2, -C(=O)-N(-H, -C1-4alkyl), and -C(=O)-N(-C,.~alkyl,
-C,_4alkyl);
alternatively, R9a taken with R9b or R9~ taken with R9a when either pair of
groups is attached to the same nitrogen atom may combine with that nitrogen
atom to form a 5-8 membered saturated, partially saturated or unsaturated
ring which contains from 0-1 additional heteroatoms selected from a group
consisting of -N, -O, S, wherein any S ring atom may be present as a -S-,
-S(=O)- or -S(=O)2 group;
J is a member selected from the group consisting of
a direct link, -CH(R")- and -CH(R")-CHZ-;
R" is a member selected from the group consisting of:
hydrogen, C,~,alkyl, CZ_6alkenyl, CZ_6alkynyl, C3_8cycloalkyl, C~alkyl-C3_
8cycloalkyl, Co_4alkylphenyl, Co~alkylnaphthyl, Co~alkylheterocyclic ring
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having from 1 to 4 hetero ring atoms selected from the group consisting of N,
O and S, CHZCOOC,~alkyl, CHzCOOC,~,alkylphenyl and
CHZCOOC,_4alkylnaphthyl;
Z is a member selected from the group consisting of:
(a) phenyl, which is independently substituted with 0-2 R'b substituents;
(b) naphthyl, which is independently substituted with 0-2 R'b
substituents; and
(c) a monocyclic or fused bicyclic heterocyclic ring system having from
5 to 10 ring atoms, wherein 1-4 ring atoms of the ring system are
selected from N, O and S, and wherein the ring system may be
substituted with 0-2 R'b substituents;
R'b is selected from:
Halo, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_
8cycloalkyl, -CN, -NOz, NR2bR3b~ SOZNR2bR3b~ SOzRzb, C 'f3, ORzb, O-CHz-
CHZ ORzb, O-CHz-COORzb, N(Rzb)-CHz-CHz-ORzb, N(-CHz-CHz-ORzb)z,
N(Rzb)-C(=O)R3b, N(Rzb)-SOz-R3b, and a 5-6 membered aromatic heterocyclic
system containing from 1-4 heteroatoms selected from N, O and S, wherein
from 1-4 hydrogen atoms on the aromatic heterocyclic system may be
independently replaced with a member selected from the group consisting of
halo, C,~alkyl, Cz_balkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co_4alkylC3_
gcycloalkyl, -CN and -NOz;
Rzb and R3b are independently selected from the group consisting of:
H, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_gcycloalkyl,
Co~,alkylC3_8cycloalkyl,
Co~alkylphenyl and Co_4alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~alkyl,
Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co_4a1ky1C3_gcycloalkyl, -CN and
-NOz
L is selected from:
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H~ -CN~ C~-O~yzR~3~ ~Cgz)nWzR~3~ C~-yz~yzR~s~ yzRis~ OR'z,
-~12C~-~12~~12RI3' and NR'zC(=NR'z)-R's;
n is an integer of 0-2;
R'z and R'3 are independently selected from:
hydrogen, -OR'4, _NR'4R'S, C,~alkyl, Co~alkylphenyl, Co~alkylnaphthyl,
COOC,_4alkyl, COO-Co~alkylphenyl and COO-Co~alkylnaphthyl, wherein
from 1-4 hydrogen atoms on the ring atoms of the phenyl and naphthyl
moieties may be independently replaced with a member selected from the
group consisting of halo, C,_4alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl,
Co_4alkylC3_8cycloalkyl, -CN, and -NOz;
R'4 and R'S are independently selected from:
H, C,~,alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_gcycloalkyl,
Co~,alkylC3_$cycloalkyl,
Co~alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~,alkyl,
Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_8cycloalkyl, -CN, and
-NOz~
and all pharmaceutically acceptable isomers, salts, hydrates, solvates and
prodrug derivatives thereof.
In certain aspects of this invention, compounds are provided which are useful
as diagnostic reagents. In another aspect, the present invention includes
pharmaceutical compositions comprising a pharmaceutically effective amount of
the
compounds of this invention and a pharmaceutically acceptable carrier. In yet
another aspect, the present invention includes methods comprising using the
above
compounds and pharmaceutical compositions for preventing or treating disease
states characterized by undesired thrombosis or disorders of the blood
coagulation
process in mammals, or for preventing coagulation in biological samples such
as, for
example, stored blood products and samples. Optionally, the methods of this
invention comprise administering the pharmaceutical composition in combination
with an additional therapeutic agent such as an antithrombotic and/or a
thrombolytic
agent and/or an anticoagulant.
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The preferred compounds also include their pharmaceutically acceptable
isomers, hydrates, solvates, salts and prodrug derivatives.
Detailed Description of the Invention
Definitions
In accordance with the present invention and as used herein, the following
terms are defined with the following meanings, unless explicitly stated
otherwise.
The term "alkenyl" refers to a trivalent straight chain or branched chain
unsaturated aliphatic radical. The term "alkinyl" (or "alkynyl") refers to a
straight or
branched chain aliphatic radical that includes at least two carbons joined by
a triple
bond. If no number of carbons is specified alkenyl and alkinyl each refer to
radicals
having from 2-12 carbon atoms.
The term "alkyl" refers to saturated aliphatic groups including straight-
chain,
branched-chain and cyclic groups having the number of carbon atoms specified,
or if
no number is specified, having up to 12 carbon atoms. The term "cycloalkyl" as
used herein refers to a mono-, bi-, or tricyclic aliphatic ring having 3 to 14
carbon
atoms and preferably 3 to 7 carbon atoms.
As used herein, the terms "carbocyclic ring structure " and " C3_16
carbocyclic
mono, bicyclic or tricyclic ring structure" or the like are each intended to
mean
stable ring structures having only carbon atoms as ring atoms wherein the ring
structure is a substituted or unsubstituted member selected from the group
consisting
of: a stable monocyclic ring which is aromatic ring ("aryl") having six ring
atoms; a
stable monocyclic non-aromatic ring having from 3 to 7 ring atoms in the ring;
a
stable bicyclic ring structure having a total of from 7 to 12 ring atoms in
the two
rings wherein the bicyclic ring structure is selected from the group
consisting of ring
structures in which both of the rings are aromatic, ring structures in which
one of the
rings is aromatic and ring structures in which both of the rings are non-
aromatic; and
a stable tricyclic ring structure having a total of from 10 to 16 atoms in the
three
rings wherein the tricyclic ring structure is selected from the group
consisting of
ring structures in which three of the rings are aromatic, ring structures in
which two
of the rings are aromatic and ring structures in which three of the rings are
non-
aromatic. In each case, the non-aromatic rings when present in the monocyclic,
bicyclic or tricyclic ring structure may independently be saturated, partially
saturated
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or fully saturated. Examples of such carbocyclic ring structures include, but
are not
limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, adamantyl,
cyclooctyl,
[3.3.0]bicyclooctane, [4.3.0]bicyclononane, [4.4.0]bicyclodecane (decalin),
2.2.2]bicyclooctane, fluorenyl, phenyl, naphthyl, indanyl, adamantyl, or
tetrahydronaphthyl (tetralin). Moreover, the ring structures described herein
may be
attached to one or more indicated pendant groups via any carbon atom which
results
in a stable structure. The term "substituted" as used in conjunction with
carbocyclic
ring structures means that hydrogen atoms attached to the ring carbon atoms of
ring
structures described herein may be substituted by one or more of the
substituents
indicated for that structure if such substitutions) would result in a stable
compound.
The term "aryl" which is included with the term "carbocyclic ring structure"
refers to an unsubstituted or substituted aromatic ring, substituted with one,
two or
three substituents selected from loweralkoxy, loweralkyl, loweralkylamino,
hydroxy,
halogen, cyano, hydroxyl, mercapto, vitro, thioalkoxy, carboxaldehyde,
carboxyl,
carboalkoxy and carboxamide, including but not limited to carbocyclic aryl,
heterocyclic aryl, and biaryl groups and the like, all of which may be
optionally
substituted. Preferred aryl groups include phenyl, halophenyl,
loweralkylphenyl,
napthyl, biphenyl, phenanthrenyl and naphthacenyl.
The term "arylalkyl" which is included with the term "carbocyclic aryl"
refers to one, two, or three aryl groups having the number of carbon atoms
designated, appended to an alkyl group having the number of carbon atoms
designated. Suitable arylalkyl groups include, but are not limited to, benzyl,
picolyl,
naphthylinethyl, phenethyl, benzyhydryl, trityl, and the like, all of which
may be
optionally substituted.
As used herein, the term "heterocyclic ring" or "heterocyclic ring system" is
intended to mean a substituted or unsubstituted member selected from the group
consisting of stable monocyclic ring having from 5-7 members in the ring
itself and
having from 1 to 4 hetero ring atoms selected from the group consisting of N,
O and
S; a stable bicyclic ring structure having a total of from 7 to 12 atoms in
the two
rings wherein at least one of the two rings has from 1 to 4 hetero atoms
selected
from N, O and S, including bicyclic ring structures wherein any of the
described
stable monocyclic heterocyclic rings is fused to a hexane or benzene ring; and
a
stable tricyclic heterocyclic ring structure having a total of from 10 to 16
atoms in
the three rings wherein at least one of the three rings has from 1 to 4 hetero
atoms
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selected from the group consisting of N, O and S. Any nitrogen and sulfur
atoms
present in a heterocyclic ring of such a heterocyclic ring structure may be
oxidized.
Unless indicated otherwise the terms "heterocyclic ring" or "heterocyclic ring
system" include aromatic rings, as well as non-aromatic rings which can be
saturated, partially saturated or fully saturated non-aromatic rings. Also,
unless
indicated otherwise the term "heterocyclic ring system" includes ring
structures
wherein all of the rings contain at least one hetero atom as well as
structures having
less than all of the rings in the ring structure containing at least one
hetero atom, for
example bicyclic ring structures wherein one ring is a benzene ring and one of
the
rings has one or more hetero atoms are included within the term "heterocyclic
ring
systems" as well as bicyclic ring structures wherein each of the two rings has
at least
one hetero atom. Moreover, the ring structures described herein may be
attached to
one or more indicated pendant groups via any hetero atom or carbon atom which
results in a stable structure. Further, the term "substituted" means that one
or more
of the hydrogen atoms on the ring carbon atoms) or nitrogen atoms) of the each
of
the rings in the ring structures described herein may be replaced by one or
more of
the indicated substituents if such replacements) would result in a stable
compound.
Nitrogen atoms in a ring structure may be quaternized, but such compounds are
specifically indicated or are included within the term "a pharmaceutically
acceptable
salt" for a particular compound. When the total number of O and S atoms in a
single
heterocyclic ring is greater than 1, it is preferred that such atoms not be
adjacent to
one another. Preferably, there are no more that 1 O or S ring atoms in the
same ring
of a given heterocyclic ring structure.
Examples of monocylic and bicyclic heterocylic ring systems, in alphabetical
order, are acridinyl, azocinyl, benzimidazolyl, benzofuranyl,
benzothiofuranyl,
benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl,
benzisoxazolyl, benzisothiazolyl, benzimidazalinyl, carbazolyl, 4aH-
carbazolyl,
carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-
dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl,
imidazolinyl, imidazolyl, 1H-indazolyl, indolinyl, indolizinyl, indolyl, 3H-
indolyl,
isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl,
isoquinolinyl
(benzimidazolyl), isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl,
octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl,
1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl,
pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
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phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl,
pteridinyl,
purinyl, pyranyl, pyrazinyl, pyroazolidinyl, pyrazolinyl, pyrazolyl,
pyridazinyl,
pryidooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl,
pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl,
4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl,
tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadazinyl,
1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-
thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl,
thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl,
1,3,4-triazolyl
and xanthenyl. Preferred heterocyclic ring structures include, but are not
limited to,
pyridinyl, furanyl, thienyl, pyrrolyl, pyrazolyl, pyrrolidinyl, imidazolyl,
indolyl,
benzimidazolyl, 1H-indazolyl, oxazolinyl, or isatinoyl. Also included are
fused ring
and spiro compounds containing, for example, the above heterocylic ring
structures.
As used herein the term "aromatic heterocyclic ring system" has essentially
the same definition as for the monocyclic and bicyclic ring systems except
that at
least one ring of the ring system is an aromatic heterocyclic ring or the
bicyclic ring
has an aromatic or non-aromatic heterocyclic ring fused to an aromatic
carbocyclic
ring structure.
The terms "halo" or "halogen" as used herein refer to Cl, Br, F or I
substituents. The term "haloalkyl", and the like, refer to an aliphatic carbon
radicals
having at least one hydrogen atom replaced by a Cl, Br, F or I atom, including
mixtures of different halo atoms. Trihaloalkyl includes trifluoromethyl and
the like
as preferred radicals, for example.
The term "methylene" refers to -CH2-.
The term "pharmaceutically acceptable salts" includes salts of compounds
derived from the combination of a compound and an organic or inorganic acid.
These compounds are useful in both free base and salt form. In practice, the
use of
the salt form amounts to use of the base form; both acid and base addition
salts are
within the scope of the present invention.
"Pharmaceutically acceptable acid addition salt" refers to salts retaining the
biological effectiveness and properties of the free bases and which are not
biologically or otherwise undesirable, formed with inorganic acids such as
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hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric
acid and
the like, and organic acids such as acetic acid, propionic acid, glycolic
acid, pyruvic
acid, oxalic acid, malefic acid, malonic acid, succinic acid, fumaric acid,
tartaric acid,
citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid,
ethanesulfonic acid, p-toluenesulfonic acid, salicyclic acid and the like.
"Pharmaceutically acceptable base addition salts" include those derived from
inorganic bases such as sodium, potassium, lithium, ammonium, calcium,
magnesium, iron, zinc, copper, manganese, aluminum salts and the like.
Particularly
preferred are the ammonium, potassium, sodium, calcium and magnesium salts.
Salts derived from pharmaceutically acceptable organic nontoxic bases include
salts
of primary, secondary, and tertiary amines, substituted amines including
naturally
occurnng substituted amines, cyclic amines and basic ion exchange resins, such
as
isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine,
ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine,
arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine,
ethylenediamine, glucosamine, methylglucamine, theobromine, purines,
piperizine,
piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly
preferred
organic nontoxic bases are isopropylamine, diethylamine, ethanolamine,
trimethamine, dicyclohexylamine, choline, and caffeine.
"Biological properly" for the purposes herein means an in vivo effector or
antigenic function or activity that is directly or indirectly performed by a
compound
of this invention that are often shown by in vitro assays. Effector functions
include
receptor or ligand binding, any enzyme activity or enzyme modulatory activity,
any
carrier binding activity, any hormonal activity, any activity in promoting or
inhibiting adhesion of cells to an extracellular matrix or cell surface
molecules, or
any structural role. Antigenic functions include possession of an epitope or
antigenic site that is capable of reacting with antibodies raised against it.
In the compounds of this invention, carbon atoms bonded to four non-
identical substituents are asymmetric. Accordingly, the compounds may exist as
diastereoisomers, enantiomers or mixtures thereof. The syntheses described
herein
may employ racemates, enantiomers or diastereomers as starting materials or
intermediates. Diastereomeric products resulting from such syntheses may be
separated by chromatographic or crystallization methods, or by other methods
known in the art. Likewise, enantiomeric product mixtures may be separated
using
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the same techniques or by other methods known in the art. Each of the
asymmetric
carbon atoms, when present in the compounds of this invention, may be in one
of
two configurations (R or S) and both are within the scope of the present
invention.
Preferred Embodiments
In a preferred embodiment, the present invention provides a compound of the
formula I:
A-Y-D-E-G-J-Z-L
wherein:
A is selected from:
(a) phenyl, which is independently substituted with 0-2 R' substituents;
(b) a monocyclic or fused bicyclic heterocyclic ring system having from 5 to
10 ring atoms, wherein 1-4 ring atoms of the ring system are selected from
N, O and S, and wherein the ring system may be substituted with 0-2 R'
substituents;
(c) naphthyl, which is independently substituted with 0-2 R' substituents;and
(d) C,-C6 alkyl; C3-C8-cycloalkyl;
(e) NRZR3, -C(=NRZ)NRzR3, -NRZC(=NRz)NRZR3, -C(=NRz)R4, and
~2~(=~2)-R3
R' is selected from:
Halo, C,_4alkyl, CZ_6alkenyl, Cz_balkynyl, C3_gcycloalkyl, Co~alkylC3_
8cycloalkyl,-CN, -NO2, -(CHZ)mNRzR3, -C(=O)NRZR3, -C(=NRZ)NRZR3,
-NRZC(=NRZ)NRZR3, -C(=NRz)R4 and NRZC(=NRZ)-R3, -SOZNRZR3, -SOZR2,
-CF3, -OR2, and a 5-6 membered aromatic heterocyclic system containing
from 1-4 heteroatoms selected from N, O and S, wherein from 1-4 hydrogen
atoms on the aromatic heterocyclic system may be independently replaced
with a member selected from the group consisting of halo, C,-C4 alkyl, -CN
C,~alkyl, CZ_6alkenyl, CZ_6allcynyl, C3_8cycloalkyl, Co_4alky1C3_8cycloalkyl
and
-NO2;
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Rz and R3 are independently selected from the group consisting of:
H, -OR'a, -NR'aR's, C,_aalkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co_
aalkylC3_8cycloalkyl, COOC,_aalkyl, COO-Co~alkylphenyl Co_aalkylphenyl
and Co~,alkylnaphthyl, wherein from 1-4 hydrogen atoms on the ring atoms
of the phenyl and naphthyl moieties may be independently replaced with a
member selected from the group consisting of halo, C,~alkyl, Cz_balkenyl, Cz_
6alkynyl, C3_8cycloalkyl, Co_aalkylC3_8cycloalkyl, -CN, and -NOz;
m is an integer of 0-2;
Y is a member selected from the group consisting of
a direct link, -C(=O)-, -N(Ra)-, -C(=O)-N(Ra)-, -N(Ra)-C(=O)-, -SOz-, -O-,
-SOz-N(Ra)-, -N(Ra)-SOz-, -C(=NRa), -C(=S)-, -CHz-, -CHZN(Ra)-;
Ra is selected from:
H, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_gcycloalkyl,
Co~alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~,alkyl,
Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_8cycloalkyl, -CN, and
_NOz~.
D is a direct link or is a member selected from the group consisting o~
(a) phenyl, which is independently substituted with 0-2 R'a substituents;
(b) naphthyl, which is independently substituted with 0-2 R'a substituents;
and
(c) a monocyclic or fused bicyclic heterocyclic ring system having from 5 to
10 ring atoms, wherein 1-4 ring atoms of the ring system are selected
from N, O and S, and wherein the ring system may be substituted
with 0-2 R'a substituents;
R'a is selected from:
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Halo, C,~alkyl, Cz_balkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_
8cycloalkyl, -CN, -NOz, (CHz)",NRzaR3a, SOzNRzaR3a, SOZRza, CF3, ORza, and
a S-6 membered aromatic heterocyclic system containing from 1-4
heteroatoms selected from N, O and S, wherein from 1-4 hydrogen atoms on
the aromatic heterocyclic system may be independently replaced with a
member selected from the group consisting of halo, C,~,alkyl, Cz_balkenyl, Cz_
6alkynyl, C3_8cycloalkyl, Co~,alkylC3_8cycloalkyl, -CN and -NOz.
m is an integer of 0-2;
Rza and R3a are independently selected from the group consisting of:
H, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_$cycloalkyl,
Co_QalkylC3_8cycloalkyl,
Co_4alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~alkyl,
Cz_6alkenyl, Cz_balkynyl, C3_8cycloalkyl, C~alkylC3_8cycloalkyl, -CN and
-NOz;.
E is a member selected from the group consisting of
-N(RS)-C(=O)_, -C(=O)_N(RS)-, -N(Rs)_C(=O)_N(R6)-, -SOz_N(RS)-,
-N(RS)-SOz-N(R6)- and N(RS)-SOz-N(R6)-C(=O)-;
RS and R6 are independently selected from:
H, C,_4alkyl, Cz_balkenyl, Cz_6alkynyl, C3_scycloalkyl,
Co~alkylC3_8cycloalkyl,
Co_4alkylphenyl, Co~alkylnaphthyl, C~alkylheteroaryl, C,_4alkylCOOH and
C,_4alkylCOOC,,~alkyl, wherein from 1-4 hydrogen atoms on the ring atoms
of the phenyl, naphthyl and heteroaryl moieties may be independently
replaced with a member selected from the group consisting of halo, C,_4alkyl,
Cz_balkenyl, Cz_balkynyl, C3_8cycloalkyl, Co~alkylC3_gcycloalkyl, -CN and
-NOz
G is selected from:
-CR'Rg- and -CR'aRga-CbRgb
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Wherein R', Rg, R'a, Rga, R'~ and R8b are each independently a member selected
from
the group consisting of
hydrogen, halo, -C,_balkyl, haloalkyl, -CN, -NOZ, -CZ_balkenyl, -Cz
6alkynyl, -C3_8cycloalkyl, -Co~alkyl-C3_g-cycloalkyl, -Co~,alkyl-CN, -Co
4alkyl-NO2, -Co_4alkyl-O-R9, -Co_Qalkyl-S-R9, -Co,~alkyl-S(=O)2-R9,
-Co_4alkyl-S(O)-R9, -Co_4alkyl-C(=O)-OR9, -Co~alkyl-C(=O)-N(R9a, R9b)
-Co_4alkyl-C(=O)-R9, -Co~alkyl-N(R9a, R9b), -
4alkyl-N(-R9a)-C(=O)-R9b), -Co~alkyl-N(-R9a)-C(=O)-R9b, -Co_
4alkyl-N(-R9a)-C(=O)-N(-R9b), -Co~alkyl-N(-R9a)-S(=O)2-R9b, -Co_
4alkyl-S(=O)2-N(R9a, R9b), -Co~alkyl-S(=O)2-R9, -Co_
4alkyl-P(=O)(-OR9a)(-OR9b), -Co~,alkyl-N(-R9)-P(=O)(-OR9a)(-OR9b), -Co_
4alkyl-phenyl, -Co~,alkyl-naphthyl, -Co~alkyl-heterocyclic ring system
containing from 1-4 heteroatoms selected from the group consisting of
O, N and S, wherein the heterocyclic ring system is a 5-6 membered
monocyclic ring or a 8-12 membered bicyclic ring, and wherein 0-4
hydrogen atoms of the phenyl ring, the naphthyl ring carbon and the
heterocyclic ring system are replaced by a member selected from the
group consisting of -C,~alkyl, haloalkyl, halo, -CN, -NO2, -OR9°, -
SR9°,
-S(O)R9~~ -C(=~)-OR9~~ -C(=O)-N(-R9~~ R9a)~ -C(=O)-R9~~ -N(R9~~ R9a)
_N(_R9~)-C(=~)-R9a~ -N(-R9~)-C(=O)-OR9a~ -N(-R9~)-C(=O)-N(-H~ R9a)
-N(-R9°)-SOZ-R9a, -SOZ-N(-R9', -R9a), -SOZ-R9c; or one of R', R8, R'a,
Rga,
R'~ and R8b can combine with a nitrogen on the E group to form a 5-7
membered heterocyclic ring containing a 0-3 additional heteroatoms
selected from the group consisting of O, N and S; or R'a and R'~ on
adjacent carbons combine to form a 3-6 membered carbocyclic ring;
R'~ and R8b combine to form alkylidene groups, such as HZC=, C,_
4a1ky1CH=, (C,~alkyl)ZC=, PhCH=;
R9, R9a, R9b, R9' and R9a are each independently a member selected from the
group
consisting of
H, halo -C,_6alkyl, -CZ_balkenyl, -CZ_balkynyl, -C3_8cycloalkyl, -Co~alkyl-C3_
gcycloalkyl, -CHZCHzOH, -CHZCHZ-O-CH3, -Co~alkylphenyl, -Co_
4alkylheterocycle wherein the heterocycle may be a 5-6 membered ring, and
wherein from 0-4 hydrogen atoms from the ring atoms of the phenyl and
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heterocycle groups may be independently replaced with a member selected
from the group consisting of halo, -C,_4alkyl, -CZ_6alkenyl, -CZ_6alkynyl,
-C3_8cycloalkyl, -Co~alkyl-C3_8cycloalkyl, -CN, -NOZ, -C(=O)-OH, -C(=O)-O-
C,_4alkyl, -C(=O)-NH2, -C(=O)-N(-H, -C1-4alkyl), and -C(=O)-N(-C,_4alkyl,
-C,_4alkyl);
alternatively, R9a taken with R9b or R9' taken with R9d when either pair of
groups is attached to the same nitrogen atom may combine with that nitrogen
atom to form a 5-8 membered saturated, partially saturated or unsaturated
ring which contains from 0-1 additional heteroatoms selected from a group
consisting of -N, -O, S, wherein any S ring atom may be present as a -S-,
-S(=O)- or -S(=O)z- group;
J is a member selected from the group consisting of:
a direct link, -CH(R" )- and -CH(R" )-CHz-;
R" is a member selected from the group consisting of
hydrogen, C,_4alkyl, CZ_6alkenyl, Cz_balkynyl, C3_gcycloalkyl, Co~,alkyl-C3_
8cycloalkyl, Co~alkylphenyl, Co~alkylnaphthyl, Co_4alkylheterocyclic ring
having from 1 to 4 hetero ring atoms selected from the group consisting of N,
O and S, CHZCOOC,_Qalkyl, CHZCOOC,~,alkylphenyl and
CHZCOOC, _4alkylnaphthyl;
Z is a member selected from the group consisting of:
(a) phenyl, which is independently substituted with 0-2 R'b substituents;
(b) naphthyl, which is independently substituted with 0-2 R'b substituents;
and
(c) a monocyclic or fused bicyclic heterocyclic ring system having from 5 to
10 ring atoms, wherein 1-4 ring atoms of the ring system are selected
from N, O and S, and wherein the ring system may be substituted
with 0-2 R'b substituents;
R'b is selected from:
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Halo, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_
8cycloalkyl, -CN, -NOz, NRzbR3b, SOzNR2bR3b~ SOzRzb~ CF3, ORzb, O-CHz-
CHz-ORzb, O-CHz-COORzb, N(Rzb)-CHz-CHz-ORzb, N(-CHz-CHz-ORzb)z,
N(Rzb)-C(=O)R3b, N(Rzb)-SOz-R3b, and a 5-6 membered aromatic heterocyclic
system containing from 1-4 heteroatoms selected from N, O and S, wherein
from 1-4 hydrogen atoms on the aromatic heterocyclic system may be
independently replaced with a member selected from the group consisting of
halo, C,~alkyl, Cz_6alkenyl, Cz_6alkynyl, C3_8cycloalkyl, Co~alkylC3_
8cycloalkyl, -CN and -NOz;
IO Rzb and R3b are independently selected from the group consisting of:
H, C,~alkyl, Cz_6alkenyl, Cz_balkynyl, C3_8cycloalkyl, Co~alkylC3_gcycloalkyl,
Co~alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,_4alkyl,
Cz_6alkenyl, Cz_6alk3myl, C3_gcycloalkyl, Co~alkylC3_gcycloalkyl, -CN and
-NOz;
L is selected from:
H~ -CN~ C~-O~yzR~s~ ~CHz)nWzR~3~ C~_~12~~12R13~ ~rzR~s~ yzRi3~
OR'z, -NR'zC(°NR'z)NR'zR's, and NR'zC(=NR'z)-R~3;
n is an integer of 0-2;
R'z and R'3 are independently selected from:
hydrogen, -OR'4, -NR'4R'S, C,_4alkyl, Co~alkylphenyl, Co_4alkylnaphthyl,
COOC,~alkyl, COO-Co~,alkylphenyl and COO-Co~alkylnaphthyl, wherein
from 1-4 hydrogen atoms on the ring atoms of the phenyl and naphthyl
moieties may be independently replaced with a member selected from the
group consisting of halo, C,~alkyl, Cz_6alkenyl, Cz_balkynyl, C3_8cycloalkyl,
Co~alkylC3_8cycloalkyl, -CN, and -NOz;
R'4 and R'S are independently selected from:
H, C,_4alkyl, Cz_balkenyl, Cz_6alkynyl, C3_8cycloalkyl,
Co~,alkylC3_gcycloalkyl,
Co~alkylphenyl and Co~alkylnaphthyl, wherein from 1-4 hydrogen atoms on
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the ring atoms of the phenyl and naphthyl moieties may be independently
replaced with a member selected from the group consisting of halo, C,~alkyl,
CZ_6alkenyl, CZ_balkynyl, C3_8cycloalkyl, Co~alkylC3_$cycloalkyl, -CN, and
-NOz;
and all pharmaceutically acceptable isomers, salts, hydrates, solvates and
prodrug derivatives thereof.
In certain aspects of this invention, compounds are provided which are useful
as diagnostic reagents. In another aspect, the present invention includes
pharmaceutical compositions comprising a pharmaceutically effective amount of
the
compounds of this invention and a pharmaceutically acceptable Garner. In yet
another aspect, the present invention includes methods comprising using the
above
compounds and pharmaceutical compositions for preventing or treating disease
states characterized by disorders of the blood coagulation process in mammals,
or
for preventing coagulation in stored blood products and samples. Optionally,
the
1 S methods of this invention comprise administering the pharmaceutical
composition in
combination with an additional therapeutic agent such as an antithrombotic
and/or a
thrombolytic agent and/or an anticoagulant.
The preferred compounds also include their pharmaceutically acceptable
isomers, hydrates, solvates, salts and prodrug derivatives.
In a further preferred embodiment, the present invention provides a
compound according to the formula I:
A-Y-D-E-G-J-Z-L
wherein:
W~ ~~/71512 CA 02374820 2001-11-20 PCT/~15~0/14207
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A is a member selected from the group consisting of:
S02NH2 S02NHMe S02Me CN CONH2 CH2NH2
/ \ / \ / \ / \ / \ / \
S02NH2 S02Me
CH2NMe2 N02 / \
/ \ / \ / \ Me-NON- Me-N~N-
V
CI CI
Me~N N Me~N/~N- NH O NH 02 ~NH
CNH C ~/ ~ S
H2N
N~ \ N~ \ ~ ~ N ~ N- N/ \
N
Me H
. N , ~ ~-- ,~ C ~-
N C ~ ~~ ~S H NMe
O
Me
NH NH NH NH NH
HO~ MeO~ H2N
N- ' N_ \ N~ HsCHN~N~ PhHN~N~
H H H H H
F
O NH ~ O NH \ N O NH
O' ' ~ O~N~ N~
H H I O
Y is a member selected from the group consisting of
S a direct link, -C(=O}-; -N(-CH3}-; -N(CH3)-CHZ-; -C(=NH)-, -CHz-, -C(=S)-, -
NH-,
and -SOz-;
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D is a member selected from the group consisting of:
CI F B r
/ \ / \ / ~ / \ / \ / \
- -C
Me F F CI F
/ \ / \ / \ / \ /-\ /
N N
F
F F
Br
F
/ \ / \ / \ / \ CI and -N
N S S S S
or A-Y-D is a member selected from the group consisting of
R1a NH2 R1a NH2 Rya NH2 Rya NH2
~N ! ~ ~N i ~ ~N i \ ~N
I / / / NJ / N / O
H
~a R~ ~ NH2
R'la NH2 Rya NH2 R NI-12
W ~N W ~N ~~ vN I / ~N
I
I/ I/ ~N / S
Wherein R'a is selected from:
hydrogen, Cl, F, Br, Me, OMe, NO2, C02H, CN, C(=O)NH2, and C(=O)OMe;
E is a member selected from the group consisting of:
-N(-H)-C(=O)-and -C(=O)-N(-H)-;
G is -CR'aRga-CbR8b ;
wherein R'a, R$a, R'~ and R8b are independently a member selected from the
group
consisting of:
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hydrogen, F, Cl, Br, -OH, -NO2, -CN, -C,~alkyl, haloalkyl, -OR9, -CHZOR9,
-S(=O)2-R9, -CHZS(=O)2-R9, -C(=O)-OR9, -CHZC(=O)-OR9, -C(=O)-N(R9a, R9b),
-CHZC(=O)-N(R9a, R9b), -N(R9a, R9b), -CHzN(R9a, R9b), -N(R9a)-C(=O)-R9b),
phenyl,
benzyl, -Co_Zalkyl-heterocyclic ring system containing from 1-4 heteroatoms
selected
from the group consisting of O, N and S, wherein the heterocyclic ring system
is a 5-
6 membered monocyclic ring; wherein the phenyl ring and heterocyclic ring are
substituted by a member selected from the group consisting of CH3, halo, -CN,
-NO2, -OMe, -COZH, -COZMe;
or R'~ and R8b combine to form CH2 , (CH3)zC=, PhCH=;
R9, R9a and R9b are independently selected from:
hydrogen, -C,_4alkyl, haloalkyl, phenyl, benzyl; or R9a and R9b may combine
with that nitrogen atom to which they are attached to form a 5-6 membered
ring which contains from 0-1 additional heteroatoms selected from a group
consisting of -N, -O, S;
J is a member selected from the group consisting of:
a direct link, -CHZ-;
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Z-L is a member selected from the group consisting of:
OH
\ \ \ \
I/ I/ I/ I/ ~/ I/
~F
HZN NH HzN OH HzN NH HzN NHMe HzN NH HzN NH
z
F ~~ HN~COZMe
\ F \ F \
I / ~ ~ I ~ I / I / I /
H2N ~O
HpN NH HzN NH HpN NH HzN NH HzN NH
HN~COZH NHz OH
\ \ \
\ / \ \ / N /
/ / /
H N NH HzN \N~ HzN \N I Hz ~N I HZN NJ HzN S
z
NH S \ \ \
I ~ HN - / / O I /
\ - NH
./
N N
HzN I N HZN I / N HzN H2N HZN
\ \
H \
I / H ~ \N / O / I F I / O /
\ N
/ HzN ~ \ D~ Hz Ni \/\N/
NH HZN N I
z
In a further preferred embodiment, the present invention provides a compound
according to the formula I:
A-Y-D-E-G-J-Z-L
wherein:
A is a member selected from the group consisting of:
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S02NH2 S02NHMe S02NHBu(t) S02Me / \ / \
/ \ / \ / \ / \
CN CONH2
/ \ / \ CH2NH2 CH2NMe2
/ \ / \
/ \ / \
N02 NC H2NOC H2NH2C
H3N
N/ \ N~\ N/ \ / \ / \ ~N- ~N-
N N N N- N
~N- ~N- N- O N- 02 N- Me-N~N-
a
NH2 NH2
- N- N N N N N Me-N N-
v
-N C~ CN CN Co C
H2N H ME
H
N N O S HsC
H Me
Y is a member selected from the group consisting of:
a direct link, -C(=O)-; -N(-CH3)-; -N(CH3)-CHz-; -C(=NH)-, -CHZ-, -C(=S)-, -NH-
,
and -SOZ-;
D is a member selected from the group consisting of
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CI F B r
/ ~ / ~ / N /
- --
Me F F CI F
/ ~ /
N N
F
F F
Br
F
\ ~ \ ~ \ ~ \ CI and -N J--
N S S g S
E is a member selected from the group consisting of
-N(-H)-C(=O)-and -C(=O)-N(-H)-;
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G is a member selected from the group consisting of:
H H
HO ~ H F ~ H HO CI CH3 HO OHCH3
OCH3
HO H3~ CH3 F CI CH3 HO ~ Ph HO CH3
H3C I ~N H3C
HO CH3 F N HO F F CH3
CI ~CH3
F
HO F
O .N.
N HN ~N N- HO N-N F F
O
y
O CH3 OCH3 H3C
CH3 CF3
HN
O
H3C02C N ~N H3C02C
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J is a direct link;
Z-L is a member selected from the group consisting of:
H
\ \ \ \
I/ I/ I/ I/ / /
-F
HzN NH Hz ~ N HzN ~ N HpN ~ N
z H N NH
OH NHz NHMe H N NH z
F ~~ HN~COzMe
\ \ \ \ F \ F /
I/ I/ I/ / F /
H N NH HzN NH HzN O
HpN NH HzN NH HZN NH z
HN~COzH NHz H
\ \ \
I \ I / I / I / I / N I /
/ O O O
H N NH HzN \N HzN \N HZN \N HzN HZN S
z
NH \ \ \
/ / H I/ / I/
\ ~NH - O
N
N
HzN / HzN ~ ~ N HzN HZN HZN
- _ \ \ I\
HN F
I / H I ~N I / O / I I / O / I
/ HzN ~N y ~ Hz ~Ni NON/
NHz HZN N I
O
S
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In a further preferred embodiment, the present invention provides a compound
according to the formula I:
A-Y-D-E-G-J-Z-L
wherein
A is a member selected from the group consisting o~
S02NH2 S02Me
\ / \ ~N\ CN- ~N-
N NH NH
HO. ~ H2N,
N NMe H H
Me
Y is a member selected from the group consisting of:
a direct link, -C(=O)-; -N(-CH3)-; -N(CH3)-CHZ-; -C(=NH)-, -CHZ-, -C(=S)-,
-NH-, and -SOZ ;
D is a member selected from the group consisting of:
CI F Br
/ ~ / \ / \ / \ / \ -N
N
E is a member selected from the group consisting o~
-N(-H)-C(=O)-and -C(=O)-N(-H)-;
G is a member selected from the group consisting o~
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H H
HO ~ H F ~ H HO CI CHs HO OHCHs
F
HO H C CHs HO HO~CH / \3
C / \I
~ i /~ O
N- HO N-N ~N
J is a direct link;
Z-L is a member selected from the group consisting of
HN~COZMe OH
I ~ F
H2N ~NH H2N ~ N H N NH H2N NH HzN ~ NH
OH z
NHz
I ~ ~ ~ S
O . O I \-J
H2N N H2N N H2N N
I
I F
O ~ I
I
. J~O~ HzN N~O~N~N~
H2N N
O
and all pharmaceutically acceptable isomers, salts, hydrates, solvates and
prodrug
derivatives thereof.
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The following non-limiting table illustrates representative compounds of the
present invention:
Table of Preferred Compounds
F F F F F
F
A_B_D_N O I \ A_B_D_N O I / A_B_D_N O I \
\
OH I
H2N NH H2N N HZN N
HO F CH3 HO F CH3 HO F CH3
A_B_D_N \ A_B_D_N I \ A_B_D_N I \
O
I O w .OH O I
HZN NH H2N N HZN N
HO N ~ HO N ~ HO N
A-g-p-N I \ A-B-D-N I \ A-g-p-N I \
O ~ O / O
I
HZN NH HzN N'OH HZN N~
- ~ ~N
~N
H
A-B-D N I \ A_g_p. A B D N O I \ \
O / I ]
HzN N-
HZN NH '
A_B_D_N \ A_B_D_N \ A_B_
O I / O I /
,OH
HZN NH H2N N
~N O O N O ~N O
A_B_p_N \ A-B-D-N \ A-B-D-N I W
o I / O I / p I \
H N H2N ~N-OH HZN N~
2 NH
Wherein:
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A is a member selected from the group consisting of:
S02NH2 S02Me
\ / \ ~N\ ~N- O\~~N-
N NH NH
HO~ ~ H2N
N NMe H H
Me
Y is a member selected from the group consisting of
a direct link, -C(=O)-; -N(-CH3)-; -N(CH3)-CHz-; -C(=NH)-, -CHZ-, -C(=S)-, -NH-
,
and -SOZ-;
D is a member selected from the group consisting of
CI F Br
/ \ / \ / \ / \ / \ -N
N
This invention also encompasses all pharmaceutically acceptable isomers,
salts, hydrates and solvates of the compounds of the invention. In addition,
the
compounds can exist in various isomeric and tautomeric forms, and all such
forms
are meant to be included in the invention, along with pharmaceutically
acceptable
salts, hydrates and solvates of such isomers and tautomers.
The compounds of this invention may be isolated as the free acid or base or
converted to salts of various inorganic and organic acids and bases. Such
salts are
1 S within the scope of this invention. Non-toxic and physiologically
compatible salts
are particularly useful although other less desirable salts may have use in
the
processes of isolation and purification.
A number of methods are useful for the preparation of the salts described
above and are known to those skilled in the art. For example, the free acid or
free
base form of a compound of one of the formulas above can be reacted with one
or
more molar equivalents of the desired acid or base in a solvent or solvent
mixture in
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which the salt is insoluble, or in a solvent like water after which the
solvent is
removed by evaporation, distillation or freeze drying. Alternatively, the free
acid or
base form of the product may be passed over an ion exchange resin to form the
desired salt or one salt form of the product may be converted to another using
the
same general process.
Prodrug Derivatives of Compounds
This invention also encompasses prodrug derivatives of the compounds
contained herein. The term "prodrug" refers to a pharmacologically inactive
derivative of a parent drug molecule that requires biotransformation, either
spontaneous or enzymatic, within the organism to release the active drug.
Prodrugs
are variations or derivatives of the compounds of this invention which have
groups
cleavable under metabolic conditions. Prodrugs become the compounds of the
invention which are pharmaceutically active in vivo, when they undergo
solvolysis
under physiological conditions or undergo enzymatic degradation. Prodrug
compounds of this invention may be called single, double, triple etc.,
depending on
the number of biotransformation steps required to release the active drug
within the
organism, and indicating the number of functionalities present in a precursor-
type
form. Prodrug forms often offer advantages of solubility, tissue
compatibility, or
delayed release in the mammalian organism (see, Bundgard, Design of Prodrugs,
pp.
7-9, 21-24, Elsevier, Amsterdam 1985 and Silverman, The Organic Chemistry of
Drug Design and Drug Action, pp. 352-401, Academic Press, San Diego, CA,
1992).
Prodrugs commonly known in the art include acid derivatives well known to
practitioners of the art, such as, for example, esters prepared by reaction of
the
parent acids with a suitable alcohol, or amides prepared by reaction of the
parent
acid compound with an amine, or basic groups reacted to form an acylated base
derivative. Moreover, the prodrug derivatives of this invention may be
combined
with other features herein taught to enhance bioavailability.
As mentioned above, the compounds of this invention find utility as
therapeutic agents for disease states in mammals which have disorders of
coagulation such as in the treatment or prevention of unstable angina,
refractory
angina, myocardial infarction, transient ischemic attacks, thrombotic stroke,
embolic
stroke, disseminated intravascular coagulation including the treatment of
septic
shock, deep venous thrombosis in the prevention of pulmonary embolism or the
treatment of reocclusion or restenosis of reperfused coronary arteries.
Further, these
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compounds are useful for the treatment or prophylaxis of those diseases which
involve the production and/or action of factor Xa/prothrombinase complex. This
includes a number of thrombotic and prothrombotic states in which the
coagulation
cascade is activated which include but are not limited to, deep venous
thrombosis,
pulmonary embolism, myocardial infarction, stroke, thromboembolic
complications
of surgery and peripheral arterial occlusion.
Accordingly, a method for preventing or treating a condition in a mammal
characterized by undesired thrombosis comprises administering to the mammal a
therapeutically effective amount of a compound of this invention. In addition
to the
disease states noted above, other diseases treatable or preventable by the
administration of compounds of this invention include, without limitation,
occlusive
coronary thrombus formation resulting from either thrombolytic therapy or
percutaneous transluminal coronary angioplasty, thrombus formation in the
venous
vasculature, disseminated intravascular coagulopathy, a condition wherein
there is
1 S rapid consumption of coagulation factors and systemic coagulation which
results in
the formation of life-threatening thrombi occurnng throughout the
microvasculature
leading to widespread organ failure, hemorrhagic stroke, renal dialysis, blood
oxygenation, and cardiac catheterization.
The compounds of the invention also find utility in a method for inhibiting
the coagulation biological samples, which comprises the administration of a
compound of the invention.
The compounds of the present invention may also be used in combination
with other therapeutic or diagnostic agents. In certain preferred embodiments,
the
compounds of this invention may be coadministered along with other compounds
typically prescribed for these conditions according to generally accepted
medical
practice such as anticoagulant agents, thrombolytic agents, or other
antithrombotics,
including platelet aggregation inhibitors, tissue plasminogen activators,
urokinase,
prourokinase, streptokinase, heparin, aspirin, or warfarin. The compounds of
the
present invention may act in a synergistic fashion to prevent reocclusion
following a
successful thrombolytic therapy andlor reduce the time to reperfusion. These
compounds may also allow for reduced doses of the thrombolytic agents to be
used
and therefore minimize potential hemorrhagic side-effects. The compounds of
this
invention can be utilized in vivo, ordinarily in mammals such as primates,
(e.g.
humans), sheep, horses, cattle, pigs, dogs, cats, rats and mice, or in vitro.
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The biological properties of the compounds of the present invention can be
readily characterized by methods that are well known in the art, for example
by the
in vitro protease activity assays and in vivo studies to evaluate
antithrombotic
efficacy, and effects on hemostasis and hematological parameters, such as are
illustrated in the examples.
Diagnostic applications of the compounds of this invention will typically
utilize formulations in the form of solutions or suspensions. In the
management of
thrombotic disorders the compounds of this invention may be utilized in
compositions such as tablets, capsules or elixirs for oral administration,
suppositories, sterile solutions or suspensions or injectable administration,
and the
like, or incorporated into shaped articles. Subjects in need of treatment
(typically
mammalian) using the compounds of this invention can be administered dosages
that
will provide optimal efficacy. The dose and method of administration will vary
from subject to subject and be dependent upon such factors as the type of
mammal
being treated, its sex, weight, diet, concurrent medication, overall clinical
condition,
the particular compounds employed, the specific use for which these compounds
are
employed, and other factors which those skilled in the medical arts will
recognize.
Formulations of the compounds of this invention are prepared for storage or
administration by mixing the compound having a desired degree of purity with
physiologically acceptable carriers, excipients, stabilizers etc., and may be
provided
in sustained release or timed release formulations. Acceptable Garners or
diluents
for therapeutic use are well known in the pharmaceutical field, and are
described, for
example, in Remington's Pharmaceutical Sciences, Mack Publishing Co., (A.R.
Gennaro edit. 1985). Such materials are nontoxic to the recipients at the
dosages
and concentrations employed, and include buffers such as phosphate, citrate,
acetate
and other organic acid salts, antioxidants such as ascorbic acid, low
molecular
weight (less than about ten residues) peptides such as polyarginine, proteins,
such as
serum albumin, gelatin, or immunoglobulins, hydrophilic polymers such as
polyvinylpyrrolidinone, amino acids such as glycine, glutamic acid, aspartic
acid, or
arginine, monosaccharides, disaccharides, and other carbohydrates including
cellulose or its derivatives, glucose, mannose or dextrins, chelating agents
such as
EDTA, sugar alcohols such as mannitol or sorbitol, counterions such as sodium
and/or nonionic surfactants such as Tween, Pluronics or polyethyleneglycol.
Dosage formulations of the compounds of this invention to be used for
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therapeutic administration must be sterile. Sterility is readily accomplished
by
filtration through sterile membranes such as 0.2 micron membranes, or by other
conventional methods. Formulations typically will be stored in lyophilized
form or
as an aqueous solution. The pH of the preparations of this invention typically
will
be 3-11, more preferably 5-9 and most preferably 7-8. It will be understood
that use
of certain of the foregoing excipients, earners, or stabilizers will result in
the
formation of cyclic polypeptide salts. While the preferred route of
administration is
by injection, other methods of administration are also anticipated such as
orally,
intravenously (bolus and/or infusion), subcutaneously, intramuscularly,
colonically,
rectally, nasally, transdermally or intraperitoneally, employing a variety of
dosage
forms such as suppositories, implanted pellets or small cylinders, aerosols,
oral
dosage formulations and topical formulations such as ointments, drops and
dermal
patches. The compounds of this invention are desirably incorporated into
shaped
articles such as implants which may employ inert materials such as
biodegradable
polymers or synthetic silicones, for example, Silastic, silicone rubber or
other
polymers commercially available.
The compounds of the invention may also be administered in the form of
liposome delivery systems, such as small unilamellar vesicles, large
unilamellar
vesicles and multilamellar vesicles. Liposomes can be formed from a variety of
lipids, such as cholesterol, stearylamine or phosphatidylcholines.
The compounds of this invention may also be delivered by the use of
antibodies, antibody fragments, growth factors, hormones, or other targeting
moieties, to which the compound molecules are coupled. The compounds of this
invention may also be coupled with suitable polymers as targetable drug
carriers.
Such polymers can include polyvinylpyrrolidinone, pyran copolymer, polyhydroxy-
propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or
polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore,
compounds of the invention may be coupled to a class of biodegradable polymers
useful in achieving controlled release of a drug, for example polylactic acid,
polyglycolic acid, copolymers of polylactic and polyglycolic acid, polyepsilon
caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,
polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block
copolymers of hydrogels. Polymers and semipermeable polymer matrices may be
formed into shaped articles, such as valves, stems, tubing, prostheses and the
like.
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Therapeutic compound liquid formulations generally are placed into a
container having a sterile access port, for example, an intravenous solution
bag or
vial having a stopper pierceable by hypodermic injection needle.
Therapeutically effective dosages may be determined by either in vitro or in
vivo methods. For each particular compound of the present invention,
individual
determinations may be made to determine the optimal dosage required. The range
of
therapeutically effective dosages will be influenced by the route of
administration,
the therapeutic objectives and the condition of the patient. For injection by
hypodermic needle, it may be assumed the dosage is delivered into the body's
fluids.
For other routes of administration, the absorption efficiency must be
individually
determined for each compound by methods well known in pharmacology.
Accordingly, it may be necessary for the therapist to titer the dosage and
modify the
route of administration as required to obtain the optimal therapeutic effect.
The
determination of effective dosage levels, that is, the dosage levels necessary
to
achieve the desired result, will be readily determined by one skilled in the
art.
Typically, applications of compound are commenced at lower dosage levels, with
dosage levels being increased until the desired effect is achieved.
The compounds of the invention can be administered orally or parenterally in
an effective amount within the dosage range of about 0.1 to 100 mg/kg,
preferably
about 0.5 to SO mg/kg and more preferably about 1 to 20 mg/kg on a regimen in
a
single or 2 to 4 divided daily doses and/or continuous infusion.
Typically, about 5 to 500 mg of a compound or mixture of compounds of this
invention, as the free acid or base form or as a pharmaceutically acceptable
salt, is
compounded with a physiologically acceptable vehicle, Garner, excipient,
binder,
preservative, stabilizer, dye, flavor etc., as called for by accepted
pharmaceutical
practice. The amount of active ingredient in these compositions is such that a
suitable dosage in the range indicated is obtained.
Typical adjuvants which may be incorporated into tablets, capsules and the
like are binders such as acacia, corn starch or gelatin, and excipients such
as
microcrystalline cellulose, disintegrating agents like corn starch or alginic
acid,
lubricants such as magnesium stearate, sweetening agents such as sucrose or
lactose,
or flavoring agents. When a dosage form is a capsule, in addition to the above
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materials it may also contain liquid carriers such as water, saline, or a
fatty oil.
Other materials of various types may be used as coatings or as modifiers of
the
physical form of the dosage unit. Sterile compositions for injection can be
formulated according to conventional pharmaceutical practice. For example,
dissolution or suspension of the active compound in a vehicle such as an oil
or a
synthetic fatty vehicle like ethyl oleate, or into a liposome may be desired.
Buffers,
preservatives, antioxidants and the like can be incorporated according to
accepted
pharmaceutical practice.
Preparation of Compounds
The compounds of the present invention may be synthesized by either solid
or liquid phase methods described and referenced in standard textbooks, or by
a
combination of both methods. These methods are well known in the art. See,
Bodanszky, "The Principles of Peptide Synthesis", Hafner, et al., Eds.,
Springer-
Verlag, Berlin, 1984.
Starting materials used in any of these methods are commercially available
from chemical vendors such as Aldrich, Sigma, Nova Biochemicals, Bachem
Biosciences, and the like, or may be readily synthesized by known procedures.
Reactions are carried out in standard laboratory glassware and reaction
vessels under reaction conditions of standard temperature and pressure, except
where
otherwise indicated.
During the synthesis of these compounds, the functional groups of the amino
acid derivatives used in these methods are protected by blocking groups to
prevent
cross reaction during the coupling procedure. Examples of suitable blocking
groups
and their use are described in "The Peptides: Analysis, Synthesis, Biology",
Academic Press, Vol. 3 (Gross, et al., Eds., 1981) and Vol. 9 (1987), the
disclosures
of which are incorporated herein by reference.
Non-limiting exemplary synthesis schemes are outlined directly below, and
specific steps are described in the Examples. The reaction products are
isolated and
purified by conventional methods, typically by solvent extraction into a
compatible
solvent. The products may be further purified by column chromatography or
other
W~ ~~/71512 CA 02374820 2001-11-20 PCT/US00/142~7
-41 -
appropriate methods.
h 1
CHO CF3 _
+ OJ KN(Me3Si)2 I \ C02CHg
~F3C~0,~~0~ 18-crown-6, THF
CN O O CN
AIMe3, DCM F
i v ~ i NH2
S02NHtBu
O
~ ~ / H ~ CN
S02NHtBu F
1.HCI, methanol
2.NH40Ac, methanol, reflux
O
N ~ I NH2
H
NH
S02NH2 F
H2, 10%Pd/C, methanol
O i
/ ~ N ~ I NH2
H NH
S02NH2 F
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chem 2
HsC HsC O H
I \ O CI OtBu KOtBu I ~ OtBu
+ ~ ~ o
CN O HOtBu CN ,
NaOH
SO2NHtBu
SOzNHtBu
O H N H2N \ / \ / HsC O H
O \ / \ / I ~ OH
1' ~ O
CN BOP CN
1) NHZOH
2) AczO / HOAc
3) HZ / Pd-C SO NH
HO H 2 2
H
O S02NHtBu I ~ O N \ / \ /
H3C H H
O N \ / \ / T~ H2N NH +
HZN NH H3C HO H SOzNH2
H
I ~ OHO N \ / \ /
1 ) MeOH / HCl (g) HZN NH
2) NH40Ac
H CCi OH SOzNHz Me0 OH SOZNH2
HC _ _
H H
N \ / \ / + I ~ o N \ / \ /
O
HzN NH H2N NH
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_43_
Scheme3
0
O N w
H + HO N 'O AcONa/AcZOH I ~ - O
~H ~ O
CN CN
H2N
/ \
A1(Me)3 S02NHtBu
DCM
/ \
H~ 1) NHzOH'HC1/Et3N/EtOH O
N ~ 2) Ac20/AcOH
O' _NHO 3) Pd-C/HZ/MeOH
~NHO
HN NH2 / \ NC / \
S02NHtBu
SOZNHtBu
/ \ / \
TFA
H of
y N
O~ NHO
HN NHz / \
SOzNHz
/-\
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Scheme 4
OH , OTf , CN
Pd2(dba)3
I TfzO/TEA/DMAP \ I dPPE~(~2 I ~ I MeMgBr
I N J MeOH I N ~ D~ N J Et O
O H O H
O C1CHZCOZtBu
I ~-OtBu SM NaOH ~ I ~-OH
I a w O MeOH I w O
N ~ tBuOH I
N N
O H 1) MCPBA
N 2) TsCI/pyridine
BOP/TEA I lT
DMF w O / \ 3) NHZCHZCHzOH
I , ~ S02NHtBu
N / \
O H CI OH
H
i I ~N H
H~ ~ I O N I
O / \ S02NHtBu % S02NH2
N NH2 MeOH I '~
/ \ N- _NH2
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cheme 5
B(OH)2
R ~O~ (CF3S02)20 R ~O~ ~ CN
O~ ~O DIEA / DCM CF3S02~0'/ ~O Pd(Ph3P)4
K3P04 / dioxane
BuHtN
S,O
_ O' _
R ~ O~ H2N ~ ~ ~ ~ R ~ NH ~
O ~ O S02NHtBu
AIMe3, DCM
CN CN
1. NCI, MeOH
2. NH40Ac, MeOH
R / NH
O S02NH2
/ NH2
NH
H2, 10% Pd/C
MeOH
R NH
O S02NH2
NH2
NH
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cheme 6
O
O
Me0 ~ LiBH4 HO I ~ IBX, DMS~ H
OMe ~ OMe
OMe CN
CN CN
H2N NHtBu
O=S=O
Me0 ~
(CF3CH20)2P(O)CH2C02Me
O I ~ OMe
KN(SiMe3)2, 18~rown-6
CN AIMe3, DGM
1. HCI, MeOH
O home 2. NH40Ac, MeOH
S02NHtH.i CN
O ~ OMe H2, 10% Pd/C
S02NH2
HN NH2 MeOH
O ~ OMe
S02N H2
HN NH2
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cheme 7
F
F O F
LDA, DMF ~ ~ ~ ~ ~ ~ N
THF H I / O
S02NH2
CN CN HN NH2
O
H3C I ~ NBS, benzoyl peroxide Br ~ \ Me3N=O H
F CCI / F CHCI3 ~ F
CN CN
CN
O / F
S02NH2
HN NH2
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cheme 8
O
/ \
I \ H ~ AcONa/Ac20H
I J ~ OEt ~(Me)3
N~-OEt I DCM
CN O ~ O
CN
/ N
N I,
O' _ N H 1 ) HC1(g)/MeOH O N H
NC 2) NH40Ac/MeOH HN NH / \
/ \ 2
S02NHtBu S02NH2
/ \ / \
/ \
Pd/C HZ I ~ ~ N
MeOH
O~ NH
HN NH2 / \
S02NH2
/ \
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cheme 9
10
Pd2(dba)3 ~ S02NHtBu
S02NHtBu Br ~ Cs2C03 / P(tBu)3
w ~ + ~ i w
B(OH)2 H dioxane I / N>
H
i w CN , SOZNHtBu 1 ) HCI (9)
CN
H02C I / w ~ ~ I EtOAc / MeOH
BOP / DIEA / DMF ~ N 2) NH40Ac / 0
3) TFA
O
S02NH2 NH , SOZNH2
NHZ HZ ~ I i
10% Pd/C ~ ~ ~ NH
N / CH30H N
O O ~ ~NFi2
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Scheme 10
Br ~ CHO OCH3 o Br ~ ~ N H2S04
+ H2N~OCH
P
CH30 OCH3
Br I ~ ~N I ~ ~N MC~ Br ' ~ ~N+O ~+ I ~ ~N+,O
. . ~+ . . ) . . . ,
Br Br
CI CI CI
POC13 Br ~ ~ ~ N I ~ ~ N separate Br I ~ ~ N
. ~+
0
Br
OPh N Hz
PhOH Br I ~ ~ N NH40Ac Br ' ~ ~ N (BochO
KOH / 0 ~ ~ D ~ ~ DIEA / DMAP
Boc~N,Boc Boc,N.Boc
Br I ~ ~ N 1 ) n-BuLi / THF /-100 °C OHC I ~ ~ N
. . 2)DMF/THF . .
Boc. N. Boc ~ SiMe3 Boc~ N, Boc
( n ~-
OHC ~ ~ N \CF3 O/2 O _ . w ~ N
KN(SiMe3)2 / 18-crown-6 / THF M~Si~ I / .
O O
1)TBAF/THF
~ S02NHtBu
I ~ ~ , S02NHtBu Boc.N,Boc
I
NHz w I . I . I ~ ~ N TFA
HATU / DIEA / DMF ~ N O ~
H
SOZNHz NH / S02NHz NH
2 I z
. / w ~N Hz _ ~ . ~ ~N
10% Pd/C ~
O ~ CH30H H O
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Scheme 11
CHO CF3 _
CH3 KN M Si I \ \C02CH3
( ~ )z_
i
f 3C~0 O Ow 18-crown-6, THF
CN ~ CN
O
AIMe3, DCM
r v ~ ~ NHz
SOZNHtBu
O
~ ~ / H ~ CN
SOZNHtBu
1.HCI, methanol
2.NH40Ac, methanol, reflux
O
/ ~ ~ I NHz
~ N
H NH
S02NHz
Hz, 10%Pd/C, methanol
O i
/ ~ w I NHz
~ N
H NH
S02NHz
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Scheme 12
OTf
HCHO \
~ N ~ ~ OEt
co2Et K2C03, Bn4NI co2Et Pd(II), TEA \ ~ o
N
H Pd-C
2
S02NHtBu
S02NHtBu H2N
AlMe3
N
1 ) mCPBA
2) TsCI / Pyridine
3) ethanolamine
4) TFA
S02NH2
N ~
i~ O
~N NH2
Compositions and Formulations
The compounds of this invention may be isolated as the free acid or base or
converted to salts of various inorganic and organic acids and bases. Such
salts are
within the scope of this invention. Non-toxic and physiologically compatible
salts
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are particularly useful although other less desirable salts may have use in
the
processes of isolation and purification.
A number of methods are useful for the preparation of the salts described
above and are known to those skilled in the art. For example, reaction of the
free
acid or free base form of a compound of the structures recited above with one
or
more molar equivalents of the desired acid or base in a solvent or solvent
mixture in
which the salt is insoluble, or in a solvent like water after which the
solvent is
removed by evaporation, distillation or freeze drying. Alternatively, the free
acid or
base form of the product may be passed over an ion exchange resin to form the
desired salt or one salt form of the product may be converted to another using
the
same general process.
Diagnostic applications of the compounds of this invention will typically
utilize formulations such as solution or suspension. In the management of
thrombotic disorders the compounds of this invention may be utilized in
compositions such as tablets, capsules or elixirs for oral administration,
suppositories, sterile solutions or suspensions or injectable administration,
and the
like, or incorporated into shaped articles. Subjects in need of treatment
(typically
mammalian) using the compounds of this invention can be administered dosages
that
will provide optimal efficacy. The dose and method of administration will vary
from subject to subject and be dependent upon such factors as the type of
mammal
being treated, its sex, weight, diet, concurrent medication, overall clinical
condition,
the particular compounds employed, the specific use for which these compounds
are
employed, and other factors which those skilled in the medical arts will
recognize.
Formulations of the compounds of this invention are prepared for storage or
administration by mixing the compound having a desired degree of purity with
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physiologically acceptable carriers, excipients, stabilizers etc., and may be
provided
in sustained release or timed release formulations. Acceptable carriers or
diluents
for therapeutic use are well known in the pharmaceutical field, and are
described, for
example, in Remington 's Pharmaceutical Sciences, Mack Publishing Co., (A.R.
Gennaro edit. 1985). Such materials are nontoxic to the recipients at the
dosages
and concentrations employed, and include buffers such as phosphate, citrate,
acetate
and other organic acid salts, antioxidants such as ascorbic acid, low
molecular
weight (less than about ten residues) peptides such as polyarginine, proteins,
such as
serum albumin, gelatin, or immunoglobulins, hydrophilic polymers such as
polyvinalpyrrolidinone, amino acids such as glycine, glutamic acid, aspartic
acid, or
arginine, monosaccharides, disaccharides, and other carbohydrates including
cellulose or its derivatives, glucose, mannose or dextrins, chelating agents
such as
EDTA, sugar alcohols such as mannitol or sorbitol, counterions such as sodium
and/or nonionic surfactants such as Tween, Pluronics or polyethyleneglycol.
Dosage formulations of the compounds of this invention to be used for
therapeutic administration must be sterile. Sterility is readily accomplished
by
filtration through sterile membranes such as 0.2 micron membranes, or by other
conventional methods. Formulations typically will be stored in lyophilized
form or
as an aqueous solution. The pH of the preparations of this invention typically
will
be between 3 and 11, more preferably from 5 to 9 and most preferably from 7 to
8.
It will be understood that use of certain of the foregoing excipients,
carriers, or
stabilizers will result in the formation of cyclic polypeptide salts. While
the
preferred route of administration is by inj ection, other methods of
administration are
also anticipated such as intravenously (bolus and/or infusion),
subcutaneously,
intramuscularly, colonically, rectally, nasally or intraperitoneally,
employing a
variety of dosage forms such as suppositories, implanted pellets or small
cylinders,
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aerosols, oral dosage formulations and topical formulations such as ointments,
drops
and dermal patches. The compounds of this invention are desirably incorporated
into shaped articles such as implants which may employ inert materials such as
biodegradable polymers or synthetic silicones, for example, Silastic, silicone
rubber
or other polymers commercially available.
The compounds of this invention may also be administered in the form of
liposome delivery systems, such as small unilamellar vesicles, large
unilamellar
vesicles and multilamellar vesicles. Liposomes can be formed from a variety of
lipids, such as cholesterol, stearylamine or phosphatidylcholines.
The compounds of this invention may also be delivered by the use of
antibodies, antibody fragments, growth factors, hormones, or other targeting
moieties, to which the compound molecules are coupled. The compounds of this
invention may also be coupled with suitable polymers as targetable drug
carriers.
Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxy-
propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, or
polyethyleneoxide-polylysine substituted with palmitoyl residues. Furthermore,
the
factor Xa inhibitors of this invention may be coupled to a class of
biodegradable
polymers useful in achieving controlled release of a drug, for example
polylactic
acid, polyglycolic acid, copolymers of polylactic and polyglycolic acid,
polyepsilon
caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,
polydihydropyrans, polycyanoacrylates and cross linked or amphipathic block
copolymers of hydrogels. Polymers and semipermeable polymer matrices may be
formed into shaped articles, such as valves, stems, tubing, prostheses and the
like.
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Therapeutic compound liquid formulations generally are placed into a
container having a sterile access port, for example, an intravenous solution
bag or
vial having a stopper pierceable by hypodermic injection needle.
Therapeutically effective dosages may be determined by either in vitro or in
vivo methods. For each particular compound of the present invention,
individual
determinations may be made to determine the optimal dosage required. The range
of
therapeutically effective dosages will naturally be influenced by the route of
administration, the therapeutic objectives, and the condition of the patient.
For
injection by hypodermic needle, it may be assumed the dosage is delivered into
the
body's fluids. For other routes of administration, the absorption efficiency
must be
individually determined for each inhibitor by methods well known in
pharmacology.
Accordingly, it may be necessary for the therapist to titer the dosage and
modify the
route of administration as required to obtain the optimal therapeutic effect.
The
determination of effective dosage levels, that is, the dosage levels necessary
to
achieve the desired result, will be within the ambit of one skilled in the
art.
Typically, applications of compound are commenced at lower dosage levels, with
dosage levels being increased until the desired effect is achieved.
A typical dosage might range from about 0.001 mg/kg to about 1000 mg/kg,
preferably from about 0.01 mg/kg to about 100 mg/kg, and more preferably from
about 0.10 mglkg to about 20 mg/kg. Advantageously, the compounds of this
invention may be administered several times daily, and other dosage regimens
may
also be useful.
Typically, about 0.5 to 500 mg of a compound or mixture of compounds of
this invention, as the free acid or base form or as a pharmaceutically
acceptable salt,
is compounded with a physiologically acceptable vehicle, Garner, excipient,
binder,
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preservative, stabilizer, dye, flavor etc., as called for by accepted
pharmaceutical
practice. The amount of active ingredient in these compositions is such that a
suitable dosage in the range indicated is obtained.
Typical adjuvants which may be incorporated into tablets, capsules and the
like are a binder such as acacia, corn starch or gelatin, and excipient such
as
microcrystalline cellulose, a disintegrating agent like corn starch or alginic
acid, a
lubricant such as magnesium stearate, a sweetening agent such as sucrose or
lactose,
or a flavoring agent. When a dosage form is a capsule, in addition to the
above
materials it may also contain a liquid Garner such as water, saline, a fatty
oil. Other
materials of various types may be used as coatings or as modifiers of the
physical
form of the dosage unit. Sterile compositions for injection can be formulated
according to conventional pharmaceutical practice. For example, dissolution or
suspension of the active compound in a vehicle such as an oil or a synthetic
fatty
vehicle like ethyl oleate, or into a liposome may be desired. Buffers,
preservatives,
antioxidants and the like can be incorporated according to accepted
pharmaceutical
practice.
In practicing the methods of this invention, the compounds of this invention
may be used alone or in combination, or in combination with other therapeutic
or
diagnostic agents. In certain preferred embodiments, the compounds of this
inventions may be coadministered along with other compounds typically
prescribed
for these conditions according to generally accepted medical practice, such as
anticoagulant agents, thrombolytic agents, or other antithrombotics, including
platelet aggregation inhibitors, tissue plasminogen activators, urokinase,
prourokinase, streptokinase, heparin, aspirin, or warfarin. The compounds of
this
invention can be utilized in vivo, ordinarily in mammals such as primates,
such as
humans, sheep, horses, cattle, pigs, dogs, cats, rats and mice, or in vitro.
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The preferred compounds of the present invention are characterized by their
ability to inhibit thrombus formation with acceptable effects on classical
measures of
coagulation parameters, platelets and platelet function, and acceptable levels
of
bleeding complications associated with their use. Conditions characterized by
undesired thrombosis would include those involving the arterial and venous
vasculature.
With respect to the coronary arterial vasculature, abnormal thrombus
formation characterizes the rupture of an established atherosclerotic plaque
which is
the major cause of acute myocardial infarction and unstable angina, as well as
also
characterizing the occlusive coronary thrombus formation resulting from either
thrombolytic therapy or percutaneous transluminal coronary angioplasty (PTCA).
With respect to the venous vasculature, abnormal thrombus formation
characterizes the condition observed in patients undergoing major surgery in
the
lower extremities or the abdominal area who often suffer from thrombus
formation
in the venous vasculature resulting in reduced blood flow to the affected
extremity
and a predisposition to pulmonary embolism. Abnormal thrombus formation
further
characterizes disseminated intravascular coagulopathy commonly occurs within
both
vascular systems during septic shock, certain viral infections and cancer, a
condition
wherein there is rapid consumption of coagulation factors and systemic
coagulation
which results in the formation of life-threatening thrombi occurnng throughout
the
microvasculature leading to widespread organ failure.
The compounds of this present invention, selected and used as disclosed
herein, are believed to be useful for preventing or treating a condition
characterized
by undesired thrombosis, such as (a) the treatment or prevention of any
thrombotically mediated acute coronary syndrome including myocardial
infarction,
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unstable angina, refractory angina, occlusive coronary thrombus occurring post-
thrombolytic therapy or post-coronary angioplasty, (b) the treatment or
prevention of
any thrombotically mediated cerebrovascular syndrome including embolic stroke,
thrombotic stroke or transient ischemic attacks, (c) the treatment or
prevention of
any thrombotic syndrome occurring in the venous system including deep venous
thrombosis or pulmonary embolus occurnng either spontaneously or in the
setting of
malignancy, surgery or trauma, (d) the treatment or prevention of any
coagulopathy
including disseminated intravascular coagulation (including the setting of
septic
shock or other infection, surgery, pregnancy, trauma or malignancy and whether
associated with mufti-organ failure or not), thrombotic thrombocytopenic
purpura,
thromboangiitis obliterans, or thrombotic disease associated with heparin
induced
thrombocytopenia, (e) the treatment or prevention of thrombotic complications
associated with extracorporeal circulation (e.g. renal dialysis,
cardiopulmonary
bypass or other oxygenation procedure, plasmapheresis), (f) the treatment or
1 S prevention of thrombotic complications associated with instrumentation
(e.g. cardiac
or other intravascular catheterization, intra-aortic balloon pump, coronary
stmt or
cardiac valve), and (g) those involved with the fitting of prosthetic devices.
Anticoagulant therapy is also useful to prevent coagulation of stored whole
blood and to prevent coagulation in other biological samples for testing or
storage.
Thus the compounds of this invention can be added to or contacted with any
medium
containing or suspected to contain factor Xa and in which it is desired that
blood
coagulation be inhibited, e.g., when contacting the mammal's blood with
material
such as vascular grafts, stems, orthopedic prostheses, cardiac stems, valves
and
prostheses, extra corporeal circulation systems and the like.
Without further description, it is believed that one of ordinary skill in the
art
can, using the preceding description and the following illustrative examples,
make
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and utilize the compounds of the present invention and practice the claimed
methods. The following working examples therefore, specifically point out
preferred embodiments of the present invention, and are not to be construed as
limiting in any way the remainder of the disclosure.
EXAMPLES
Example 1. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(3-
amidinophenyl)-propionamide.
S02NH2
H
N
O
H2N ~ NH
A. Preparation of N-tert-butyl benzenesulfonamide
To a solution of tert-butylamine (5.73g, 78.4mmo1) and triethylamine
(16.6m1, 119mmol) in dichloromethane (200m1) in an ice bath was added
benzenesulfonyl chloride (13.85g, 78.4mmo1) dropwise. The mixture was stirred
at
room temperature overnight. It was washed with saturated sodium carbonate
(60m1)
and brine (60m1). The organic layer was separated, and the aqueous layer was
extracted with dichloromethane (2x50m1). The combined organic extracts were
dried over magnesium sulfate. The solvent was evaporated in vacuo to give the
title
compound as a light yellowish solid (15.92g, 95%). ES-MS (M+H)+ = 214.
B. Preparation of 2-(tert-butylaminosulfonyl)-benzeneboronic acid.
To a solution of N-tert-butyl benzenesulfonamide ( 15.92g, 74.7mmo1) in
tetrahydrofuran (200m1) in an ice bath was added 1.6M n-butyllithium in hexane
(100m1, 164mmo1) dropwise over 30 minutes. The mixture remained a clear
solution. In an ice bath it was added triisopropylborate (24.1m1, 104mmo1)
dropwise. The mixture was stirred at room temperature for 3.Shrs, solution
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becoming cloudy. After it was cooled in an ice bath, 1N hydrochloride (200m1)
was
added. The mixture was stirred at room temperature overnight. It was extracted
with
ether (2x50m1). The organic extract was washed with 1N sodium hydroxide
(2x60m1). The aqueous solution was acidified to pH=1 with 6N hydrochloride,
and
then extracted with ether (2x100m1). The ether extract was dried over
magnesium
sulfate, and concentrated in vacuo to give the title compound as a while solid
(ll.Sg,
60%). ES-MS (M+H)+ = 258.
C. Preparation of 4-[(2-tent-butylaminosulfonyl)phenyl]-aniline
To a solution of 2-(tent-butylaminosulfonyl)-benzeneboronic acid (6.00g,
23.35mmo1) in 120m1 toluene was added water (16m1), isopropanol (60m1), and
NaOH (40m1, SM aqueous solution). To this were added 4-bromoaniline and
Pd(Ph3P)4. This heterogeneous mixture is then refluxed for 6hr, then stirred
at
room temperature over night before refluxing for another l.Shr. The reaction
mixture is then partitioned between water and ethyl acetate. The aqueous layer
is
extracted twice with ethyl acetate. The organic layers are then dried over
MgS04,
filtered and concentrated in vacuo. The crude residue is purified by silica
gel flash
chromatography. The desired product can be eluded with 30% ethyl acetate in
hexanes and concentrated to give an orange solid (5.06g, 16.65, 71%). ES-
MS(M+H)+=305.
D. Preparation of (Z) methyl 3-cyanocinnamate
To a solution of bis(2,2,2-trifluoroethyl)(methoxycarbonylmethyl)
phosphonate (1.00 g, 3.14 mmol) and 18-crown-6 (4.14 g, 15.7 mmol) in THF (50
mL) at -78 C, potassium bis(trimethylsilyl)amide (6.3 mL, 0.5 M in toluene,
3.15
mmol) was added dropwise. After the addition was completed, 3-
cyanobenzaldehyde
(0.412 g, 3.14 mmol) in THF (8 mL) was added. The mixture was stirred at - 78
C
for 30 min before it was quenched with aq. NH4C1. Water and ether were added.
Aqueous phase was separated, extracted with ether once more. The combined
organic solutions were dried over Na2S04, concentrated in vacuo to give a
solid,
which was purified by a silica gel column, first eluted with EtOAc/hexane
(5/95),
then with EtOAc/hexane (10/90) to give the titled compound (0.40 g) (yield:
68%).
MS 188 (M + H).
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E. Preparation of (2Z)-N-[4-(2-[(tert-butylamino)sulfonyl]phenyl)phenyl]-3-(3-
cyanophenyl)-acrylamide.
To a solution of 4-(2'-tert-butylaminosulfonylphenyl)aniline (80 mg, 0.263
mmol) in CH2C12 (4 mL) at room temperature, trimethylaluminum (0.39 mL, 2.0 M
in hexane, 0.78 mmol) was added dropwise. After the solution was stirred for
30 min
at room temperature, compound (Z) methyl 3-cyanocinnamate (50 mg, 0.267 mmol)
was added. The mixture was stirred at room temperature for 2 days. The
solution
was neutralized with 1N HCl (10 mL) to pH = 1-2. Water and CH2Cl2 were added,
and organic phase was separated, dried over Na2S04, concentrated in vacuo to
give
a yellowish soild (120 mg) (yield: 98%), which was sufficiently pure to be
used in
the following reaction. MS 482 (M + Na)
F. Preparation of (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-
amidinophenyl)-acrylamide.
To a solution of compound (2Z)-N-[4-(2-[(tert-
butylamino)sulfonyl]phenyl)phenyl]-3-(3-cyanophenyl)-acrylamide ( 120 mg,
0.261
mmol) in anhydrous MeOH cooled in ice bath, hydrogen chloride gas was bubbled
to saturation. The solution was then stirred at room temperature overnight. It
was
concentrated in vacuo, the residue was dissolved in anhydrous MeOH (4 mL). To
the
solution, NH40Ac (120 mg, 1.56 mmol) was added. The mixture was heated to
reflux for 0.5 h. It was concentrated in vacuo. The residue was purified by
HPLC
using a gradient of 5% CH3CN in H20 (containing 0.1% TFA) to 80% CH3CN
over 60 min. Fractions containing the desired product were pooled, and
lyophilized
to give a powder (50 mg) (yield: 46%). MS 421 (M + H)
G. Preparation of N- f 4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-amidinophenyl)-
propionamide.
A solution of compound (2Z)-N-[4-(2-[(tert-
butylamino)sulfonyl]phenyl)phenyl]-3-(3-cyanophenyl)-acrylamide (12 mg, 22
~mol) and Pd-C (S%, 6 mg) in MeOH (2 mL) was hydrogenated in a Parr Shaker
under 50 psi overnight. The solution was then filtered through a plug of
celite. The
filtrate was concentrated in vacuo to give the titled compound (12 mg) (yield:
99%).
MS 423 (M + H)
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Example 2. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-2-
methoxycarbonyl-3-(3-amidinophenyl)-propionamide.
S02NH2
O H
N \ / \ /
~C02Me
i
H2N NH
A. Preparation of (2E) and (2Z) tent-butyl 2-methoxycarbonyl-3-(3-cyanophenyl)-
acrylate.
To a solution of 3-cyanobenzaldehyde (0.700 g, 5.34 mmol) and t-butyl
methyl malonate (0.845 mL, 5.00 mmol) in toluene (40 mL), piperidine (0.500
mL,
5.06 mmol) was added. The mixture was heated to reflux overnight. Dean-Stark
apparatus was used to remove generated water. Ethyl acetate (50 mL) and 0.5 N
HCl
(SO mL) were added. Organic phase was separated, washed with saturated aq.
NaHC03, dried over Na2S04, concentrated in vacuo to give an oil. The oil was
dry-
packed onto a silica gel column, eluted with hexane first, then with 5% to 10%
EtOAc in hexane gradually to give the desired product as a mixture of trans
and cis
isomers (0.44 g) (yield: 31%).
B. Preparation ofN-[4-(2{[(tert-butyl)amino]sulfonyl}phenyl)phenyl]-2-
methoxycarbonyl-3-(3-cyanophenyl)-acrylamide.
Compound (2E) and (2Z) tert-butyl 2-methoxycarbonyl-3-(3-cyanophenyl)-
acrylate (0.220 g, 0.767 mmol) was dissolved in TFA (6 mL). It was allowed to
stand at room temperature for 2 h. TFA was removed in vacuo to give a solid.
The
solid was dissolved in anhydrous DMF (7 mL). To the solution, 4-(2'-tert-
butylaminosulfonylphenyl)aniline (0.242 g, 0.796 mmol) and triethylamine
(0.200
mL, 1.44 mmol) were added, followed by addition of BOP (0.416 g, 0.940 mmol).
The mixture was then stirred room temperature overnight. Water and ethyl
acetate
were added. Organic phase was separated, washed with saturated aq. NaHC03,
dried
over Na2S04, concentrated in vacuo to give a solid (0.391 g) (yield: 99%). It
was
sufficiently pure to be used in the following reaction.
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C. Preparation ofN-{4-[(2-aminosulfonyl)phenyl]phenyl}-2-methoxycarbonyl-3-(3-
amidinophenyl)-acrylamide.
To a solution of compound N-[4-(2 {[(tert-
butyl)amino]sulfonyl}phenyl)phenyl]-2-methoxycarbonyl-3-(3-cyanophenyl)-
acrylamide (0.390 g, 0.750 mmol) in anhydrous MeOH cooled in ice bath,
hydrogen
chloride gas was bubbled to saturation. The solution was then stirred at room
temperature overnight. It was concentrated in vacuo, the residue was dissolved
in
anhydrous MeOH (6 mL). To the solution, NH40Ac (0.450 g, 5.84 mmol) was
added. The mixture was heated to reflux for 0.5 h. It was concentrated in
vacuo. The
residue was purified by HPLC using a gradient of 5% CH3CN in H20 (containing
0.1% TFA) to 95% CH3CN over 90 min. Fractions containing the desired product
were pooled, and lyophilized to give a powder (60 mg) (yield: 17%). MS 479 (M
+
H)
D. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-2-methoxycarbonyl-3-(3-
amidinophenyl)-propionamide.
A solution of compound N-{4-[(2-aminosulfonyl)phenyl]phenyl}-2-
methoxycarbonyl-3-(3-amidinophenyl)-acrylamide (9 mg, 19 pmol) and Pd-C (10%,
7 mg) in MeOH (2 mL) was hydrogenated under balloon H2 overnight. The solution
was then filtered through a plug of celite. The filtrate was concentrated in
vacuo to
give the titled compound (9 mg) (yield: 99%). MS 481 (M + H)
Example 3. Preparation of N-{4-[(2-aminosulfonyl)phenyl]-2-fluorophenyl}-3-(3-
amidinophenyl)-propionamide.
0
/ ~ w I NH2
N
H NH
S02NH2 F
A. Preparation of 4-[(2-tert-butylaminosulfonyl)phenyl]-2-fluoro-aniline.
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To a solution of 2-(tert-butylaminosulfonyl)-benzeneboronic acid (2.06g,
8mmol) in toluene (60m1) was added water (4m1), 8N sodium hydroxide (8m1),
isopropanol (16m1), 2-fluoro-4-iodoaniline (3.8g, l6mmol) and
tetrakis(triphenylphosphine)palladium(0) (464mg, 0.4mmo1). The mixture was
refluxed for 3-4 hrs, cooled to room temperature, and diluted with ethyl
acetate. The
organic layer was washed with water (25m1), and dried over magnesium sulfate.
After the evaporation of the solvent in vacuo, the crude reside was purified
by silica
gel column chromatography using solvent system 20-30% ethyl acetate in hexane
as
eluent to give the title compound as a white solid (1.49g, 58%). ES-MS (M+H)+
_
323.
B. Preparation of (2Z)-N-{4-[2-(tent-butylaminosulfonyl)phenyl]-2-
fluorophenyl}-3-
(3-cyanophenyl)-acrylamide
To a solution of 4-[(2-tert-butylaminosulfonyl)phenyl]-2-fluoro-aniline
(161mg, 0.5mmol) in dichloromethane (5m1) was added 2.0M trimethylaluminum
in hexane (0.75m1, l.5mmo1). The mixture was stirred at room temperature for
30
minutes, methane gas evolved. A solution of (Z) methyl 3-cyanocinnamate (94mg,
0.5mmo1) in dichloromethane (1m1) was added. The mixture was stirred at room
temperature overnight. 1N hydrochloride was added to acidify the solution to
pH=2.
After the addition of water and dichloromethane, the organic layer was
separated,
and the aqueous layer was extracted with dichloromethane. The combined organic
extracts were dried over magnesium sulfate, and concentrated in vacuo to give
the
title compound as a yellow oil (260mg, 100%). ES-MS (M+H)+ = 478.
C. (2Z)-N-{4-[2-(aminosulfonyl)phenyl]-2-fluorophenyl}-3-(3-amidinophenyl)-
acrylamide
To a solution of (2Z)-N-{4-[2-(tent-butylaminosulfonyl)phenyl]-2-
fluorophenyl}-3-(3-cyanophenyl)-acrylamide (100mg, 0.21mmo1) in absolute
methanol (3m1) in an ice bath was saturated with hydrochloride gas for 10
minutes.
The mixture was stirred at room temperature for 3 hrs. After the evaporation
of
solvent in vacuo, the residue was dissolved in absolute methanol (3m1), and
ammonia acetate (97mg, 1.26mmo1) was added. The mixture was refluxed for 3
hrs.
The solvent was evaporated in vacuo. The crude residue was purified by RP-HPLC
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to give the title compound as a white powder (53mg, 58%). ES-MS (M+H)+ _
439.
D. Preparation of N-{4-[(2-aminosulfonyl)phenyl]-2-fluorophenyl)-3-(3-
amidinophenyl)-propionamide.
To a solution of (2Z)-N-{4-[2-(aminosulfonyl)phenyl]-2-fluorophenyl}-3-(3-
amidinophenyl)-acrylamide (30mg, 0.07mmo1) in absolute methanol (2m1) was
added 10% Pd/C (catalytic amount). The mixture was hydrogenated under balloon
for lhr. After the filtration through celite, the solvent was evaporated in
vacuo. The
residue was purified by RP-HPLC to give the compound as a white powder (25mg,
81 %). ES-MS (M+H)+ = 441.
Example 4. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-2-(2-
furylcarbonylamino)-3-(3-amidinophenyl)-propionamide
H O
N ~1
O~ N HO
HN NH2 / \
S02NH2
/ \
A. Preparation of 3-[2-(furyl-2-yl)-5-oxo-1,3-oxazolin-4-
ylidene)methyl]benzenecarbonitrile.
A mixture of 3-cyanobenzaldehyde (2.102g, 15.320mmo1), N-2-
furoylglycine (1.85g, 10.9 mmol), and sodium acetate (0.636g, 7.753mmo1) in
15m1
acetic anhydride was refluxed for 7 hours. The mixture was then cooled to room
temperature before cooling in the freezer over night. The solid was washed
with ice
cold water then filtered (0.472g, 1.788mmo1, 16%). ES-MS(M+H)+=265.
B. Preparation ofN-{4-[(2-tert-butylaminosulfonyl)phenyl]phenyl}-2-(2-
furylcarbonylamino)-3-(3-cyanophenyl)-acrylamide.
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To a solution of 4-[(2-tert-butylaminosulfonyl)phenyl]-aniline (0.152g,
O.SOOmmol) in 9m1 DCM was added trimethylaluminum (!ml, 2M solution in
hexanes, 2mmol) which was allowed to stir for'/ hour. Then 3-[(2-(2-fury!)-S-
oxo-
1,3-oxazolin-4-ylidene)methyl]benzenecarbonitrile (0.!!g, 0.417mmol) was added
S drop wise as a solution in 3m1 DCM. Three hours later 6M HCl was added drop
wise to pH=0. l Oml portions of water and DCM were also added and the aqueous
layer was extracted twice with l Oml portions of DCM. The organic layers were
dried over MgS04, filtered and concentrated in vacuo to yield the desired
product
(0.259, 0.456, 109%). ES-MS(M+H)+=569.
C. Preparation ofN-{4-[(2-tent-butylaminosulfonyl)phenyl]phenyl}-2-(2-
furylcarbonylamino)-3-(3-amidinophenyl)-propionamide.
To a solution of N-{4-[(2-tert-butylaminosulfonyl)phenyl]phenyl}-2-(2-
furylcarbonylamino)-3-(3-cyanophenyl)-acrylamide (0.259g, 0.456mmol) in 7m!
ethanol was added hydroxyamine (0.192g, 2.763mmo1) and triethyl amine
(0.762m1,
5.407mmol). This mixture was refluxed for 2 hours before it was concentrated
in
vacuo. The residue was dissolved in AcOH (5m!), then acetic anhydride (0.30m1,
3.182mmo1) was added and the mixture was allowed to stir forl.5 hours. The
mixture was concentrated in vacuo. The residue was dissolved in dry MeOH
(3m1),
5%Pd/C (22.7mg) was added. A balloon filled with hydrogen gas was fitted to
the
flask with an adapter. The flask was evacuated and backfilled with hydrogen
gas
three times before being run for 0.75 hour. The mixture was then filtered over
a bed
of celite and concentrated in vacuo. The residue was purified via Preparative
HPLC
to yield the desired product (0.075g, 0.128mmo1, 28%). ES-MS(M+H)+=588.
D. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-2-(2-
furylcarbonylamino)-3-(3-amidinophenyl)-propionamide
A solution of compound N-{4-[(2-tert-butt'laminosulfonyl)phenyl]phenyl)-
2-(2-furylcarbonylamino)-3-(3-amidinophenyl)-propionamide (0.075g, 0.128mmol)
in TFA (6m!) was stirred at room temperature for 2 hours. The mixture was
concentrated in vacuo and the residue was purified via preparative HPLC to
give the
product (0.040g, yield: 58%). ES-MS(M+H)+=532.
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Example S. Preparation of N-[4-(2-methylsulfonylphenyl)phenyl]-3-(3-
amidinophenyl)-propionamide.
S02CH3
/
O I/
H ~NHz
A. Preparation of N-{4-[(2-methylsulfonyl)phenyl]phenyl}-3-(3-cyanophenyl)-
acrylamide
To a solution of 4-(2-methylsulfonylphenyl)aniline (74.1 mg, 0.3 mmol, 1.0
equiv) in 5 mL of CHZC12 at 0°C, was added a solution of AlMe3 (2M in
hexanes,
0.7 mL, 5 equiv). After l5min, methyl 3-cyanocinnamate (56.1 mg, 1.0 equiv)
was
added. The resulting solution was stirred overnight, carefully quenched with
water,
diluted with ethyl acetate. The organic layer was dried, evaporated and
chromatographied on silica gel to give the product in 55% yield. LRMS found
for
Cz3H,9N2O3S (M+H)+: 403.1.
B. Preparation of N-{4-[(2-methylsulfonyl)phenyl]phenyl}-3-(3-amidinophenyl)-
acrylamide
1 S The compound N-{4-[(2-methylsulfonyl)phenyl]phenyl}-3-(3-cyanophenyl)-
acrylamide (25 mg) was dissolved in 5 mL of methanol . The reaction mixture
was
cooled to 0°C and HCl gas was bubbled in until saturation. The mixture
was stirred
at room temperature overnight. The solvent was evaporated and the resulting
residue
was treated with ammonium acetate and 10 ml methanol at reflux temperature for
2
h. The solvent was removed at reduced pressure and the crude pruduct was
purified
by HPLC (C18 reversed phase) eluting with 0.5% TFA in H20/CH3CN to give the
desired compound in 77% yield. LRMS found for C23H22N3~3S (T'1+H)+: 420.1.
C. Preparation of N-[4-(2-methylsulfonylphenyl~henyl]-3-(3-amidinophenyl)-
propionamide
The compound N-{4-[(2-methylsulfonyl)phenyl]phenyl}-3-(3-
amidinophenyl)-acrylamide (8 mg) and S mg of 10% Pd/C was suspended in 1 mL
of methanol . The reaction mixture was stirred under 1 atm hydrogen balloon
for 2h
and filtered. The solvent was removed at reduced pressure and the crude
product was
purified by HPLC (C18 reversed phase) eluting with 0.5% TFA in H20/CH3CN to
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give the desired compound in 63% yield. LRMS found for Cz3H24N3O3S (M+H)+:
422.1.
Example 6. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(3-amidino-4-
fluorophenyl)-propionamide.
/ \ / \ N ( w
O ~~ F
S02NH2
HN NHZ
A. Preparation of 2-fluoro-5-bromomethylbenzonitrile.
2-Fluoro-5-methyl benzonitrile (1.26g, 9.32 mmol) was mixed with NBS
(1.66 g, 9.32 mmol), benzoyl peroxide (79 mg, 0.33 mmol) in CCl4 (45mL). The
mixture was refluxed for 2.5 hrs. It was cooled to room temperature, filtered
and
concentrated in vacuo to give the title compound. ES-MS (M+H)+ = 213.1.
B. Preparation of 3-cyano-4-fluorobenzaldehyde.
To a solution of 2-fluoro-S-bromomethylbenzonitrile (9.32 mmol) in CHC13
(SO mL), was added trimethylamino N-oxide (1.7 g, 23.3 mmol). The mixture was
refluxed for 3 hrs. Water was added. The organic layer was dried over MgS04,
filtered and filtrate was concentrated in vacuo. The residue was purified by
silica gel
column chromatography using solvent system 20% EtOAc in hexane as eluant to
give the title compound. ES-MS (M+H)+ = 150.1.
C. Preparation of (Z) methyl 3-cyano-4-fluorocinnamate
To a solution of bis(2,2,2-trifluoroethyl)(methoxycarbonylmethyl)
phosphonate (0.12 mL, 0.58 mmol) and 18-crown-6 (770 mg, 2.92 mmol) in THF (5
mL) at -78°C, was added potassium bis(trimethylsilyl)amide (1.17 mL,
0.57 mmol)
dropwise. After the addition was complete, 3-cyano-4-fluorobenzaldehyde (87
mg,
0.58 mmol) in THF (2 mL) was added. The mixture was stirred at -78°C
for 1 hour.
Aqueous NH4Cl solution was added to quench the reaction. Water and EtOAc was
added to the mixture. The organic layer was dried over MgS04, filtered and
concentrated in vacuo. This was purified by silica gel column chromatography
using
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solvent system 20% EtOAc in hexane as eluant to give the title compound (85
mg,
71 %). ES-MS (M+H)+ = 206.1.
D. Preparation of (2Z)-N-[4-(2-[(tert-butylamino)sulfonyl]phenyl)phenyl]-3-(3-
cyano-4-fluorophenyl)-acrylamide
To a solution of 4-[(2-tert-butylaminosulfonyl)phenyl]-aniline (121.6 mg, 0.4
mmol) in DCM (3 mL) was added trimethylaluminum (0.6 mL, 2M in hexane)
dropwise. The reaction mixture was stirred at room temperature for 30 min.
Compound (Z) methyl 3-cyano-4-fluorocinnamate (82 mg, 0.4 mmol) in DCM (2
mL) was added dropwise. The mixture was stirred at room temperature overnight.
2N HCl was added to pH 2. Water and DCM were added. The organic layer was
dried over MgS04 and concentrated in vacuo. It was purified by silica gel
column
chromatography using solvent system 50% EtOAc in hexane as eluant to give the
title compound. ES-MS (M+Na)+ = 500.1.
E. Preparation of (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl)-3-(3-amidino-4-
fluorophenyl)-acrylamide
A solution of (2Z)-N-[4-(2-[(tert-butylamino)sulfonyl]phenyl)phenyl]-3-(3-
cyano-4-fluorophenyl)-acrylamide (99 mg, 0.208 mmol) in MeOH (10 mL) was
treated with a stream of HCl gas for 10 min. at 0°C. The resulting
solution was
capped, stirred at room temperature overnight and evaporated in vacuo. The
residue
was reconstituted in MeOH (10 mL) and the mixture was treated with NH40Ac (80
mg, 1.04 mmol). The reaction mixture was refluxed for 2 hrs. and concentrated
in
vacuo. The obtained residue was purified by RP-HPLC to give the title compound
as a white powder. ES-MS (M+H)+ = 439.1.
F. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl{-3-(3-amidino-4-
fluorophenyl)-propionamide
The compound (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-amidino-
4-fluorophenyl)-acrylamide (10 mg, 0.022 mmol) was dissolved in MeOH (5 mL)
and 10% PdIC (catalytic amount) was added. The mixture was hydrogenated under
balloon overnight, filtered through Celite to remove the catalyst and the
filtrate was
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evaporated. The obtained residue was purified by RP-HPLC to give the title
compound as a white powder. ES-MS (M+H)+ = 441.1.
Example 7. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-fluoro-5-
amidinophenyl)-propionamide.
A. Preparation of 2-fluoro-5-cyanobenzaldehyde.
F
/ \ / \
O
S02NH2
HN NH2
To a solution of LDA (2.6 mL, 2N solution in hexane, 5.2 mmol) in THF (10
mL) at -78°C, was added 4-fluorobenzonitrile in THF (10 mL) dropwise.
The
mixture was stirred at -78°C for 1 hour. To the mixture was added DMF
(0.4 mL).
The mixture was stirred at -78°C for another 1 S min., quenched rapidly
with AcOH
(2 mL) and water (10 mL), extracted with ether (50 mL). The ether extracts
were
washed with 1N HCl (10 mL), brine (10 mL), dried over MgS04, filtered and
concentrated in vacuo to give the title compound. (M+H)+ = 150.
B. Preparation of the final compound N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-
fluoro-5-amidinophenyl)-propionamide, starting from 2-fluoro-5-
cyanobenzaldehyde, was completed analogously to preparation of N-[4-(2-
aminosulfonylphenyl)phenyl]-3-(3-amidino-4-fluorophenyl)-propionamide in
Example 6. ES-MS (M+H)+ = 441.1.
Example 8. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(3-amidino-4-
methoxyphenyl)-propionamide.
/ \ / \
o i
OMe
S02NH2
HN NH2
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A. Preparation of 3-cyano-4-methoxybenzyl alcohol.
To a solution of methyl-3-cyano-4-methoxybenzoate (5g, 26.2 mmol) in THF
(50 mL) was added lithium borohydride (53 mL, 2.00M solution in THF, 105 mmol)
at room temperature. The mixture was stirred at room temperature overnight. 1N
HCl was slowly added until bubbling stopped. THF was removed in vacuo and
EtOAc and water were added. The organic layer was washed with water, saturated
NaHC03 solution, brine, dried with NazS04 and solvent evaporated in vacuo to
give
the title compound (3.7 g, 86.7%).
B. Preparation of 3-cyano-4-methoxybenzaldehyde.
To a solution of 3-cyano-4-methoxybenzyl alcohol (2g, 12.3 mmol) in DMSO
(50 mL) was added IBX (4.673g, 17.7 mmol) slowly. The mixture was stirred at
room temperature overnight. EtOAc and water were added. The formed precipitate
was removed. The organic layer was washed with 1N HCI, water, saturated
NaHC03, brine, dried over NazS04 and concentrated in vacuo. The obtained
residue
was purified by silica gel column chromatography using DCM as eluant to give
the
title compound (1.1g, 56%). ES-MS (M+H)+= 162.1.
C. Preparation of the final compound N-[4-(2-aminosulfonylphenyl)phenyl]-3-(3-
amidino-4-methoxyphenyl)-propionamide, starting from 3-cyano-4-
methoxybenzaldehyde, was completed analogously to preparation of N-[4-(2-
aminosulfonylphenyl)phenyl]-3-(3-amidino-4-fluorophenyl)-propionamide in
Example 6. ES-MS (M+H)+ = 443.1.
Example 9. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-methoxy-5-
amidinophenyl)-propionamide.
OMe
O
S02NH2
HN NH2
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The compound was prepared analogously to preparation of N-[4-(2-
aminosulfonylphenyl)phenyl]-3-(3-amidino-4-methoxyphenyl)-propionamide in
Example 8. ES-MS (M+H)+ = 443.1.
Example 10. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-(2-
methoxyethoxy)-5-amidinophenyl)-propionamide.
home
O
\ /
O
S02NH2
HN NH2
The compound was prepared analogously to preparation of N-[4-(2-
aminosulfonylphenyl)phenyl]-3-(3-amidino-4-methoxyphenyl)-propionamide in
Example 8. ES-MS (M+H)+ = 487.1.
Example 11. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-hydroxy-5-
amidinophenyl)-propionamide. MS (M+H)+ = 429.
OH
\
'-' O
S02NH2
HN NH2
Example 12. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(4-hydroxy-3-
amidinophenyl)-propionamide. MS (M+H)+ = 429.
/ \ / \ N
O ~ OH
S02NH2
HN NH2
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Example 13. Preparation of (2S) N-[4-(2-aminosulfonylphenyl)phenyl]-2-amino-3-
(3-amidinophenyl)-propionamide.
H NH2
'-' O /
S02NH2
HN NH2
A. Preparation of (2S) N-[4-(2-(tert-butylaminosulfonyl)phenyl)phenyl]-2-tert-
butoxycarbonylamino-3-(3-cyanophenyl)-propionamide
N-Boc-meta-cyano-phenylalanine (200 mg, 0.69 mmol) and 4-[(2-tert-
butylaminosulfonyl)phenyl]-aniline (210 mg, 0.69 mmol) were dissolved in DMF
(3
mL). DIEA (0.24 mL, 1.4 mmol) was added followed by the addition of the
coupling
reagent PyBOP (572 mg, 1.1 mmol). The solution was stirred at room temperature
for 12 hours. The reaction mixture was diluted in a mixture of EtOAc/H20. The
organic layer was washed with water, saturated NazC03, water, 1M KHS04, brine,
dried over MgS04, filtered and solvent evaporated to give the title compound.
ES-
MS (M+H)+ = 521.1.
B. Preparation of (2S) N-[4-(2-aminosulfonylphenyl)phenyl]-2-amino-3-(3-
amidinophenyl)-propionamide.
A solution of (2S) N-[4-(2-(tert-butylaminosulfonyl)phenyl)phenyl]-2-tert-
butoxycarbonylamino-3-(3-cyanophenyl)-propionamide (132 mg, 0.23 mmol) in
MeOH (10 mL) was treated with a stream of HCl gas for 10 min. at
0°C. The
resulting solution was capped, stirred at room temperature overnight and
evaporated
in vacuo. The residue was reconstituted in MeOH (10 mL) and the mixture was
treated with NH40Ac (540 mg, 7 mmol). The reaction mixture was refluxed for 2
hrs. and concentrated in vacuo. The obtained residue was purified by RP-HPLC
to
give the title compound as a white powder. ES-MS (M+H)+ = 438.1.
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Example 14. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-methyl-3-(3-
amidinophenyl)-propionamide.
S02NH2
H
~N \ / \ /
O
H2N NH
Part A. Ethyl 3-{[(trifluoromethyl)sulfonyl]-oxy}-2-propenoate and ethyl (Z)-3-
{[(trifluoromethyl)-sulfonyl]oxy}-2-propenoate
To a solution of ethyl acetoacetate (1.3g, lOmmol) in lOml anhydrous
dichloromethane was added triethylamine (1.46m1, 10.5mmo1). The reaction was
cooled to -78°C under argon to which trifluoromethanesulfonic anhydride
(2.96g,
10.5mmo1) was added dropwise via syringe over 5 minutes. Reaction was allowed
to
warm to room temperature and stirred over night. Next morning the reaction was
diluted with 25m1 dichloromethane, organic was washed with 2x50m1 water,
2x50m1
1N HCI, dried over magnesium sulfate, filtered and concentrated. Crude oil was
chromatographed on silica gel using 5% EtOAc in hexane as the eluent to give
1) ethyl
(E)-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-propenoate (800mg, 60%) as a clear
oil:
H'NMR (CDC13) : 1.247-1.282 (t, 3H); 2.471 (s, H); 4.155-4.209 (m, 2H); 5.912
(s, H);
and 2) ethyl (Z)-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-propenoate (450mg, 30%)
as a
clear oil: H'NMR (CDC13) : 1.247-1.283 (t, 3H); 2.131 (s, 3H); 4.18-4.233 (m,
2H);
5.736 (s, H)
Part B. Ethyl (Z) 3-(3-cyanophenyl)-2-propenoate
To a solution of ethyl (Z)-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-propenoate
(330mg, 1.25mmo1) in 5m1 anhydrous dioxane was added potassium phosphate
(398mg, 1.88mmo1), 3-cyanophenyl boronic acid (185mg, 1.25mmo1), and tetrakis
(triphenylphosphine)palladium(0) (36mg, 0.031mmol). Reaction mixture was
heated
to reflux and stirred overnight. Mixture was filtered through a pad of Celite,
diluted
with 50m1 ethyl acetate, washed with 2x50m1 water, 2x50m1 saturated brine
solution,
dried over magnesium sulfate, filtered and concentrated in vacuo. Residue was
chromatographed on silica gel using 5% EtOAc in hexane as the eluent to give
ethyl
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(Z) 3-(3-cyanophenyl)-2-propenoate (240mg, 71 %) as a clear oil after drying.
ES-MS
(M+H+): 216.05
Part C. Preparation of (2Z) N-[4-(2-aminosulfonylphenyl)phenyl]-3-methyl-3-(3-
amidinophenyl)-acrylamide.
To a solution of 2'-tert-butylaminosulfonyl-4-amino-[1,1']-biphenyl (198mg,
0.65mmol) in Sml anhydrous dichloromethane was added a solution of 2M
trimethylaluminum in hexane (0.98m1, 1.95mmol). Reaction was stirred at room
temperature for 20 minutes to which a solution of ethyl (Z) 3-(3-cyanophenyl)-
2-
propenoate (140mg, 0.65mmol) in lml anhydrous dichloromethane was added.
Reaction was stirred at room temperature overnight. Reaction was quenched with
Sml
1N HCl after which an additional 20m1 dichloromethane was added. Organic was
washed with 2x25m1 water, dried over magnesium sulfate and concentrated to
give
(2Z) N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-methyl-3-(3-cyanophenyl)-
acrylamide (200mg, 65%) as a light brown residue which was sufficiently pure
to be
used without further purification.
To a solution of (2Z) N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-
methyl-3-(3-cyanophenyl)-acrylamide (90mg, 0.19mmo1) in Sml anhydrous methanol
cooled in an ice bath was bubbled HCl gas until saturation was achieved.
Reaction was
allowed to warm to room temperature and stirred overnight. The reaction was
then
concentrated in vacuo and dried under hi vacuum. The dried residue was
dissolved in
Sml anhydrous methanol to which ammonium acetate (144mg, 2mmol) was added and
the reaction heated to reflux for 2 hours. The reaction was concentrated and
purified on
a 2x25cm Vydac C,8 HPLC column to give 3-((1Z)-1-methyl-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl}vinyl)-benzenecarboxamidine (35mg, 20%) as a
fluffy white powder after lyophilization. ES-MS (M+H+): 435.1
Part D. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-methyl-3-(3-
amidinophenyl)-propionamide
To a solution of (2Z) N-[4-(2-aminosulfonylphenyl)phenyl]-3-methyl-3-(3-
amidinophenyl)-acrylamide (Smg, O.O115mmo1) in 4m1 methanol was added 10% Pd
on carbon (2mg). Mixture was treated with SOpsi hydrogen on the PARK apparatus
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for lhr. Reaction was filtered through a pad of Celite, concentrated and
lyophilized to
give the title compound (3mg, 60%) as a fluffy white powder. ES-MS (M+H+):
437.1
Example 15. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-
trifluoromethyl-3-
(3-amidinophenyl)-propionamide.
F3C S02NH2
H
~N \ / \ /
i O
H2N NH
Part A. Ethyl (Z)-4,4,4-trifluoro-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-
butenoate
To a solution of ethyl trifluoroacetoacetate (5g, 27.2mmo1) in 20m1 anhydrous
dichloromethane was added triethylamine (5.7m1, 40.7mmol). Reaction was cooled
under argon to -78°C to which trifluoromethanesulfonic anhydride (1
1.5g, 10.5mmo1)
was added dropwise via syringe over 5 minutes. Reaction was allowed to warm to
room temperature and stirred over night. Next morning the reaction was diluted
with
25m1 dichloromethane, organic was washed with 2x50m1 water, 2x50m1 1N HCI,
dried
over magnesium sulfate, filtered and concentrated in vacuo. Crude oil was
chromatographed on silica gel using 5% EtOAc in hexane as the eluent to give
ethyl
(Z)-4,4,4-trifluoro-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-butenoate (7.7g,
90%) as a
clear light yellow oil after drying. H'NMR (CDC13) : 1.31-1.35 (t, 3H); 4.33-
4.35 (m,
2H); 6.535 (s, H).
Part B. Ethyl (2E)-3-(3-cyanophenyl)-4,4,4-trifluorobut-2-enoate
To a solution of ethyl (Z)-4,4,4-trifluoro-3-{[(trifluoromethyl)sulfonyl]-oxy}-
2-
butenoate (250mg, 0.79mmol) in Sml anhydrous dioxane was added potassium
phosphate (251mg, 1.19mmol), 3-cyanophenyl boronic acid (116mg, 0.79mmo1), and
tetrakis (triphenylphosphine)palladium(0) (23mg, 0.02mmo1). Reaction mixture
was
heated to reflux and stirred overnight. Mixture was filtered through a pad of
Celite,
diluted with SOmI ethyl acetate, washed with 2x50m1 water, 2x50m1 saturated
brine
solution, dried over magnesium sulfate, filtered and concentrated in vacuo.
Residue
was chromatographed on silica gel using 20% EtOAc in hexane as the eluent to
give
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ethyl (2E)-3-(3-cyanophenyl)-4,4,4-trifluorobut-2-enoate (150mg, 79%) as a
yellow
residue after drying. H'NMR (CDC13) 1.107-1.142 (t, 3H); 4.05-4.107 (m, 2H);
6.684
(s, H); 7.38-7.72 (m, 4H).
Part C. Preparation of (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-
trifluoromethyl-3-
(3-amidinophenyl)-acrylamide.
To a solution of 2'-tert-butylaminosulfonyl-4-amino-[1,1']-biphenyl (79mg,
0.26mmo1) in Sml anhydrous dichloromethane was added a solution of 2M
trimethylaluminum in hexane (0.39m1, 0.78mmo1). Reaction was stirred at room
temperature for 20 minutes to which a solution of ethyl (Z) 3-(3-cyanophenyl)-
4,4,4-
trifluoro-2-butenoate (70mg, 0.26mmol) in lml anhydrous dichloromethane was
added.
Reaction was stirred at room temperature overnight. Reaction was quenched with
Sml
1N HCl after which an additional 20m1 dichloromethane was added. Organic was
washed with 2x25m1 water, dried over magnesium sulfate, filtered and
concentrated to
give (2E) N-[4-(2-tent-butylaminosulfonylphenyl)phenyl]-3-trifluoromethyl-3-(3-
cyanophenyl)-acrylamide (120mg, 88%) as a yellow foam which was sufficiently
pure
to be used without further purification.
To a solution of (2E) N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-
trifluoromethyl-3-(3-cyanophenyl)-acrylamide (90mg, 0.19mmo1) in lOml 1:1
ethyl
acetate:anhydrous methanol cooled to -78°C was bubbled HCl gas until
saturation was
achieved. Reaction was placed in the refrigerator at 0°C over the
weekend. The
reaction was then concentrated in vacuo and dried under hi vacuum. The dried
methyl
imidate residue was dissolved in Sml anhydrous methanol to which ammonium
acetate
( 144mg, 2mmo1) was added and the reaction heated to reflux for 2 hours. The
reaction
was concentrated then treated with l Oml trifluoroacetic acid for 2hrs,
concentrated and
purified on a 2x25cm Vydac C,8 HPLC column to give the title compound (57mg,
47%) as a fluffy white powder after lyophilization. ES-MS (M+H+): 489.15
Part D. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-trifluoromethyl-3-
(3-
amidinophenyl)-propionamide
To a solution of (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-trifluoromethyl-
3-(3-amidinophenyl)-acrylamide (lOmg, 0.02mmo1) in 4m1 methanol was added 10%
Pd on carbon (2mg). Mixture was treated with hydrogen at 1 atmosphere under
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balloon for lhr. Reaction was filtered through a pad of Celite, concentrated
and
lyophilized to give the title compound (8mg, 82%) as a fluffy white powder. ES-
MS
(M+H+): 491.1
Example 16. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(1-
pyrazolylmethyl)-3-(3-amidinophenyl)-propionamide.
N'~
N S02NH2
H
~N \ / \ /
O
H2N NH
Part A. 3-(2-bromoacetyl) benzonitrile
To a solution of 3-acetobenzonitrile (5g, 0.0344mo1) in 45m1 glacial acetic
acid
was added pyridinium tribromide (11.3g, 0.0355mo1). Reaction was stirred at
room
temperature under argon overnight. Reaction was then quenched with a saturated
sodium sulfite solution (20m1) and extracted with 3x25m1 dichloromethane.
Combined
organic phases were washed with 2x25m1 water, dried over magnesium sulfate,
filtered
and concentrated in vacuo. Crude oil was chromatographed on silica gel using
5%
EtOAc in hexane as the eluent to give 3-(2-bromoacetyl) benzonitrile (4.5g,
58%) as a
white solid. H'NMR (CDC13) 4.371-4.403 (s, 2H); 7.613-7.664 (m, H); 7.838-
7.888
(m, H); 8.192-8.261 (m, 2H)
Part B. 3-[2-(IH 1-Pyrazolyl)acetyl]benzonitrile
To a solution of 3-(2-bromoacetyl)benzonitrile (SOOmg, 2.23mmol) in Sml
dichloromethane was added pyrazole (304mg, 4.46mmo1) and triethylamine
(0.31m1,
2.23mmo1). Reaction was stirred at room temperature over night. Reaction was
then
diluted with 20m1 dichloromethane, washed with 2x25m1 water, 2x25m1 1N HCI,
dried
over magnesium sulfate, filtered and concentrated in vacuo. Crude residue was
chromatographed on silica gel using 2.5% EtOAc in hexane to give 3-[2-(IH 1-
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pyrazolyl)acetyl]benzonitrile (330mg, 70%) as a clear oil after drying. ES-MS
(M+H+): 212.05
Part C. Methyl (E)-3-(3-cyanophenyl)-4-(1H 1-pyrazolyl)-2-butenoate
To a solution of bis(2,2,2-trifluoroethyl)(methoxycarbonylmethyl)phosphonate
(0.39m1, 1.87mmo1) in Sml anhydrous tetrahydrofuran was added a solution of 18-
crown-6 (2g, 7.8mmo1) in Sml anhydrous tetrahydrofuran. Reaction was cooled to
-78°
C to which a O.SM solution of potassium bis(trimethylsilyl)amide in toluene
(0.93m1,
1.87mmo1) was added all at once. The reaction mixture was stirred at -
78° C for 20
minutes after which a solution of 3-[2-(IH 1-pyrazolyl)acetyl]- benzonitrile
(330mg,
1.56mmo1) in Sml anhydrous tetrahydrofuran was added dropwise over several
minutes. Reaction was gradually allowed to warm to room temperature and
stirred for
5 hours. Reaction was then quenched with a saturated ammonium chloride
solution
(lOml) and extracted with 2x25m1 diethyl ether. Combined organic layers were
washed
with 2x25m1 water, 2x25m1 saturated brine solution, dried over magnesium
sulfate,
filtered and concentrated to a brown residue. Crude residue was
chromatographed on
silica gel using a gradient of 5% EtOAc in hexane containing 0.1%
triethylamine to
20% EtOAc in hexane containing 0.1 % triethylamine to give methyl (E)-3-(3-
cyanophenyl)-4-(IH 1-pyrazolyl)-2-butenoate (135mg, 32%) as a clear oil after
drying.
H'NMR (CDC13) 3.521 (s, #H); 4.98 (s, 2H); 5.694 (s, H); 6.237-6.247 (t, H);
7.296-
7.593 (m, 6H). NOE experiment confirmed the stereochemical configuration.
Part D. Preparation of (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-
trifluoromethyl-3-
(3-amidinophenyl)-acrylamide.
To a solution of 2'-tert-butylaminosulfonyl-4-amino-[l,l']-biphenyl (lOSmg,
0.34mmo1) in 4m1 anhydrous dichloromethane was added a solution of 2M
trimethylaluminum in hexane (O.SmI, 1.02mmo1). Reaction was stirred at room
temperature for 20 minutes to which a solution of methyl (E)-3-(3-cyanophenyl)-
4-(1H
1-pyrazolyl)-2-butenoate (90mg, 0.34mmo1) in lml anhydrous dichloromethane was
added. Reaction was stirred at room temperature overnight. Reaction was
quenched
with Sml 1N HCl after which an additional 20m1 dichloromethane was added.
Organic
was washed with 2x20m1 water, dried over magnesium sulfate, filtered and
concentrated to give (2E) N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-(1-
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pyrazolylmethyl)-3-(3-cyanophenyl)-acrylamide (155mg, 85%) as an off white
foam
which was sufficiently pure to be used without further purification.
To a solution of (2E) N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-(1-
pyrazolylmethyl)-3-(3-cyanophenyl)-acrylamide (155mg, 0.287mmo1) in lOml 1:1
ethyl acetate:anhydrous methanol cooled to -78°C was bubbled HCl gas
until saturation
was achieved. Reaction was allowed to warm to room temperature and stirred
overnight. The reaction was then concentrated in vacuo and dried under hi
vacuum.
The dried methyl imidate residue was dissolved in Sml anhydrous methanol to
which
ammonium acetate (77mg, lmmol) was added and the reaction heated to reflux for
2
hours. The reaction was concentrated, treated with trifluoroacetic acid (lOml)
for 2hrs,
concentrated and purified on a 2x25cm Vydac C,8 HPLC column to give the title
compound (40mg, 28%) as a fluffy white powder after lyophilization. ES-MS
(M+H+):
SO1.1
Part E. N-[4-(2-aminosulfonylphenyl)phenyl]-3-(1-pyrazolylmethyl)-3-(3-
amidinophenyl)-propionamide.
To a solution of (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-trifluoromethyl-
3-(3-amidinophenyl)-acrylamide (Smg, O.Olmmol) in 4m1 methanol was added 10%
Pd
on carbon (lmg). Mixture was treated with hydrogen at 1 atmosphere under
balloon
for lhr. Reaction was filtered through a pad of Celite, concentrated and
lyophilized to
give the title compound (Smg, 100%) as a fluffy white powder. ES-MS (M+H+):
503.1
Example 17. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-furyl)-3-
(3-
amidinophenyl)-propionamide.
O ~ S02NH2
H
~N \ / \ /
O
H2N NH
Part A. Ethyl (Z)-3-(2-furyl)-3- f [(trifluoromethyl)sulfonyl]-oxy}-2-
propenoate
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To a solution of ethyl B-oxo-3-furanpropionate (1g, 5.49mmo1) in 5m1
anhydrous dichloromethane was added triethylamine (0.847m1, 6.04mmol).
Reaction
was cooled under argon to -78°C to which trifluoromethanesulfonic
anhydride (1.02m1,
6.04mmo1) was added dropwise via syringe over 5 minutes. Reaction was allowed
to
warm to room temperature and stirred over night. Next morning the reaction was
diluted with 25m1 dichloromethane, organic was washed with 2x50m1 water,
2x50m1
1N HCI, dried over magnesium sulfate, filtered and concentrated in vacuo. The
crude
oil was chromatographed on silica gel using 20% EtOAc in hexane as the eluent
to give
ethyl (Z)-3-(2-furyl)-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-propenoate (1.6g,
93%) as a
light brown solid after drying. H'NMR (CDC13) 1.31-1.35 (t, 3H); 4.26-4.314
(m,
2H); 6.065 (s, H); 6.522 (s, H); 7.47 (s, H); 7.76 (s, H).
Part B. Ethyl (E) 3-(3-cyanophenyl)-3-(2-furyl)-2-propenoate
To a solution of ethyl (Z)-3-(2-furyl)-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-
propenoate (500mg, 1.59mmo1) in 7m1 anhydrous dioxane was added potassium
phosphate (506mg, 2.4mmo1), 3-cyanophenyl boronic acid (234mg, 1.59mmo1), and
tetrakis (triphenylphosphine)palladium(0) (46mg, 0.04mmo1). Reaction mixture
was
heated to reflux and stirred overnight. Mixture was filtered through a pad of
Celite,
diluted with 50m1 ethyl acetate, washed with 2x50m1 water, 2x50m1 saturated
brine
solution, dried over magnesium sulfate, filtered and concentrated in vacuo.
The crude
residue was chromatographed on silica gel using a gradient from 5% EtOAc in
hexane
to 10% EtOAc in hexane as the eluent to give ethyl (E) 3-(3-cyanophenyl)-3-(2-
furyl)-
2-propenoate (100mg, 24%) as a clear yellow oil after drying. H'NMR (CDC13)
1.1-
1.14 (t, 3H); 4,016-4.035 (m, 2H); 5.293 (s, H); 7.45-7.549 (m, 3H); 7.669 (m,
H). ES-
MS (M+H+): 268.05
Part C. (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-furyl)-3-(3-
amidinopheny1)-
acrylamide.
To a solution of 2'-tButylaminosulfonyl-4-amino-[1,1']-biphenyl (102mg,
0.336mmo1) in 4m1 anhydrous dichloromethane was added a solution of 2M
trimethylaluminum in hexane (0.5m1, l.Ommo1). Reaction was stirred at room
temperature for 20 minutes to which a solution of ethyl (E) 3-(3-cyanophenyl)-
3-(2-
furyl)-2-propenoate (90mg, 0.336mmo1) in lml anhydrous dichloromethane was
added.
Reaction was stirred at room temperature overnight. Reaction was quenched with
5m1
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1N HCl after which an additional 20m1 dichloromethane was added. Organic was
washed with 2x20m1 water, dried over magnesium sulfate and concentrated to
give
(2E)-N-[4-(2- { [(tert-butyl)amino] sulfonyl } phenyl)phenyl]-3 -(3 -
cyanophenyl)-3-(2-
furyl)prop-2-enamide (200mg, 112%) as a brown foam which was sufficiently pure
to
be used without further purification.
To a solution of (2E)-N-[4-(2-{[(tert-butyl)amino]sulfonyl{phenyl)phenyl]-3-
(3-cyanophenyl)-3-(2-furyl)prop-2-enamide (176mg, 0.336mmol) in lOml 1:1 ethyl
acetate:anhydrous methanol cooled to -78°C was bubbled HCl gas until
saturation was
achieved. Reaction was allowed to warm to room temperature and stirred
overnight.
The reaction was then concentrated in vacuo and dried under hi vacuum. The
dried
methyl imidate residue was dissolved in Sml anhydrous methanol to which
ammonium
acetate (144mg, 2mmol) was added and the reaction heated to reflux for 2
hours. The
reaction was concentrated, treated with trifluoroacetic acid (lOml) for 2hrs,
concentrated and purified on a 2x25cm Vydac C,$ HPLC column to give the title
compound (60mg, (37%) as a fluffy off white powder after lyophilization. ES-MS
(M+H+): 487.15
Part D. N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-furyl)-3-(3-amidinophenyl)-
propionamide.
To a solution of (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-(2-furyl)-3-(3-
amidinophenyl)-acrylamide (lOmg, 0.02mmo1) in 4m1 methanol was added 10% Pd on
carbon (2mg). Mixture was treated with hydrogen at 1 atmosphere under balloon
for
lhr. Reaction was filtered through a pad of Celite, concentrated and
lyophilized to give
the title compound (9mg, 90%) as a fluffy white powder. ES-MS (M+H+): 489.15
Example 18. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-3-methoxymethyl-
3-
(3-amidinophenyl)-propionamide.
OMe S02NH2
H
~N \ / \ /
O
H2N NH
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Part A. Methyl (Z)-4-methoxy-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-butenoate
To a solution of methyl 4-methoxy-3-oxobutanoate (5g, 34.2mmol) in 20m1
anhydrous dichloromethane was added triethylamine (5.24m1, 37.6mmol). Reaction
was cooled under argon to -78°C to which trifluoromethanesulfonic
anhydride
(10.6gm1, 37.6mmo1) was added dropwise via syringe over 5 minutes. Reaction
was
allowed to warm to room temperature and stirred over night. Next morning the
reaction was diluted with 25m1 dichloromethane, organic was washed with 2x50m1
water, 2x50m1 1N HCI, dried over magnesium sulfate, filtered and concentrated
in
vacuo. The crude oil was chromatographed on silica gel using a gradient of 5%
EtOAc
in hexane to 10% EtOAc in hexane as the eluent to give methyl (Z)-4-methoxy-3-
{[(trifluoromethyl)sulfonyl]-oxy}-2-butenoate (5.1g, 54%) as a clear colorless
oil after
drying. H'NMR (CDC13) 3.342 (s, 3H); 3.711 (s, 3H); 3.99 (s, H); 6.02 (s, H).
Part B. Methyl (E)-3-(3-cyanophenyl)-4-methoxy-2-butenoate
To a solution of methyl (Z)-4-methoxy-3-{[(trifluoromethyl)sulfonyl]-oxy}-2-
butenoate (246mg, 1.Ommo1) in Sml anhydrous dioxane was added potassium
phosphate (318mg, l.5mmo1), 3-cyanophenyl boronic acid (162mg, l.Ommol), and
tetrakis (triphenylphosphine)palladium(0) (29mg, 0.0251mmo1). Reaction mixture
was
heated to reflux and stirred overnight. Mixture was filtered through a pad of
Celite,
diluted with 20m1 ethyl acetate. Organic was washed with 2x20m1 water, 2x20m1
saturated brine solution, dried over magnesium sulfate, filtered and
concentrated in
vacuo to give methyl (E)-3-(3-cyanophenyl)-4-methoxy-2-butenoate (220mg, 75%)
as
a clear brown oil which was sufficiently pure to be used without further
purification.
ES-MS (M+H+): 232.1
Part C. (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-methoxymethyl-3-(3-
amidinophenyl)-acrylamide.
To a solution of 2'-tButylaminosulfonyl-4-amino-[l,l']-biphenyl (lOSmg,
0.35mmol) in 4m1 anhydrous dichloromethane was added a solution of 2M
trimethylaluminum in hexane (0.53m1, l.OSmmol). Reaction was stirred at room
temperature for 20 minutes to which a solution of methyl (E) 3-(3-cyanophenyl)-
4-
methoxy-2-butenoate (80mg, 0.35mmol) in lml anhydrous dichloromethane was
added. Reaction was stirred at room temperature overnight. Reaction was
quenched
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with 5m1 1N HCl after which an additional 20m1 dichloromethane was added.
Organic
was washed with 2x20m1 water, dried over magnesium sulfate and concentrated to
give
(2E)-N-[4-(2- { [(tert-butyl)amino] sulfonyl}phenyl)phenyl]-3-(3-cyanophenyl)-
4-
methoxybut-2-enamide (150mg, 85%) as a white foam after drying which was
sufficiently pure to be used without further purification.
To a solution of (2E)-N-[4-(2-{[(tert-butyl)amino]sulfonyl]phenyl)phenyl]-3-
(3-cyanophenyl)-4-methoxybut-2-enamide (150mg, 0.298mmol) in lOml 1:l ethyl
acetate:anhydrous methanol cooled to -78°C was bubbled HCl gas until
saturation was
achieved. Reaction was allowed to warm to room temperature and stirred
overnight.
The reaction was then concentrated in vacuo and dried under hi vacuum. The
dried
methyl imidate residue was dissolved in 5m1 anhydrous methanol to which
ammonium
acetate (77mg, lmmol) was added and the reaction heated to reflux for 2 hours.
The
reaction was concentrated, treated with trifluoroacetic acid (lOml) for 2hrs,
concentrated and purified on a 2x25cm Vydac C,8 HPLC column to give the title
compound (34mg, (25%) as a fluffy off white powder after lyophilization. ES-MS
(M+H+): 465.15
Part D. N-[4-(2-aminosulfonylphenyl)phenyl]-3-methoxymethyl-3-(3-
amidinophenyl)-
propionamide.
To a solution of (2E) N-[4-(2-aminosulfonylphenyl)phenyl]-3-
methoxymethyl-3-(3-amidinophenyl)-acrylamide (5mg, O.Olmmol) in 4m1 methanol
was added 10% Pd on carbon (lmg). Mixture was treated with hydrogen at 1
atmosphere under balloon for lhr. Reaction was filtered through a pad of
Celite,
concentrated and lyophilized to give the title compound (5mg, 100%) as a
fluffy white
powder. ES-MS (M+H+): 467.15
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Example 19. Preparation of N- f 4-[(2-aminosulfonyl)phenyl]phenyl}-N-
(carboxylmethyl)-3-(3-amidinophenyl)-2-fluoro-3-methylpropionamide.
F ~ 02H S02NH2
N \ / \ /
O
H2N NH
A. Ethyl 3-(3-cyanophenyl)-2-fluoro-3-methylacrylate.
To a solution of triethyl 2-fluoro-2-phosphonoacetate (0.838 mL, 4.13 mmol)
in anhydrous THF (25 mL) at -78 C, potassium bis(trimethylsilyl)amide (0.5 M
in
toluene, 10.0 mL, 5.00 mmol) was added dropwise. After 10 min following the
addition, a solution of 3-acetylbenzonitrile (0.600 g, 4.14 mmol) in THF (8
mL) was
added dropwise. The reaction mixture was stirred at -78 C for 30 min, then
removed to room temperature, and stirred at the temperature overnight. Aqueous
NH4C1 and EtOAc were added. Organic phase was separated, washed with sat.
NaCI, dried over Na2S04, concentrated in vacuo to give an oil as a mixture of
E-
and Z-isomers in a ratio of 5 : 1 (0.920g, yield: 95%), which was pure enough
to be
used in the next reaction. MS 234 (M + H).
B. N- f 4-[(2-tert-butylaminosulfonyl)phenyl]phenyl}-3-(3-cyanophenyl)-2-
fluoro-3-methylacrylamide.
To the solution of 4-(2'-tert-butylaminosulfonylphenyl)aniline (0.195 g,
0.641 mmol) in CH2Cl2 (8 mL) at room temperature, trimethylaluminum (2.0 M in
hexane, 0.96 mL, 1.92 mmol) was added dropwise. The reaction mixture was
stirred
for 15 min. A solution of ethyl 3-(3-cyanophenyl)-2-fluoro-3-methylacrylate
(0.149
g, 0.639 mmol) in CH2C12 (5 mL) was added. It was stirred overnight. 1N HCl
was
added to neutralize the solution to pH 2-3. Water and CH2C12 were added.
Organic
phase was separated, dried over Na2S04, concentrated in vacuo to give a solid
(0.290 g, yield: 92%), which was pure enough to be used in the next reaction.
MS
436 (M + H - 'Bu) and 514 (M + Na).
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C. N-{4-[(2-tert-butylaminosulfonyl)phenyl]phenyl}-N-(methoxycarbonylmethyl)-
3-(3-cyanophenyl)-2-fluoro-3-methylacrylamide
To a solution of N-{4-[(2-tert-butylaminosulfonyl)phenyl]phenyl}-3-(3-
cyanophenyl)-2-fluoro-3-methylacrylamide (230mg, 0.47mmol) in 15m1 DMF was
added cesium carbonate (460mg, 1.41mmo1) and bromomethyl acetate (355mg,
2.35mmol). The reaction mixture was stirred at room temperature for 4 hours
then
diluted with 25m1 of ethyl acetate. Organic was washed with 3x25m1 water,
3x25m1
saturated brine solution, dried over magnesium sulfate, filtered and
concentrated in
vacuo to give the title compound (230mg, 86%) as yellow foam. ES-MS (M+H+):
563.2.
D. N-{4-[(2-aminosulfonyl)phenyl]phenyl}-N-(methoxycarbonylmethyl)-3-(3-
amidinophenyl)-2-fluoro-3-methylacrylamide
To a solution of N-{4-[(2-tent-butylaminosulfonyl)phenyl]phenyl}-N-
(methoxycarbonylmethyl)-3-(3-cyanophenyl)-2-fluoro-3-methylacrylamide (230mg,
0.408mmo1) in lOml 1:1 ethyl acetate:anhydrous methanol cooled to -78°C
was
bubbled HCl gas until saturation was achieved. Reaction was allowed to warm to
room
temperature and stirred 18 hours. The reaction was then concentrated in vacuo
and
dried under hi vacuum. The dried methyl imidate residue was dissolved in 5m1
anhydrous methanol to which ammonium acetate (115mg, l.5mmo1) was added and
the
reaction heated to reflux for 2 hours. The reaction was then concentrated and
purified
on a 2x25cm Vydac C,8 HPLC column to give the title compound (150mg, 70%) as a
fluffy white powder after lyophilization. ES-MS (M+H+): 525.2.
E. N-{4-[(2-aminosulfonyl)phenyl]phenyl}-N-(carboxylmethyl)-3-(3-
amidinophenyl)-2-fluoro-3-methylacrylamide
To a solution of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-N-
(methoxycarbonylmethyl)-3-(3-amidinophenyl)-2-fluoro-3-methylacrylamide
(100mg,
0.19mmo1) in 5m1 methanol was added a 0.5N lithium hydroxide solution (1m1,
0.5mmo1). The reaction was stirred at room temperature for 4 hours then
concentrated
and purified on a 2x25cm Vydac C,8 HPLC column to give the title compound
(70mg,
71%) as a fluffy white powder after lyophilization. ES-MS (M+H+): 511.1.
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F. N-{4-[(2-aminosulfonyl)phenyl]phenyl{-N-(carboxylmethyl)-3-(3-
amidinophenyl)-
2-fluoro-3-methylpropionamide.
To a solution of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-N-(carboxylmethyl)-
3-(3-amidinophenyl)-2-fluoro-3-methylacrylamide (9mg) in 4m1 methanol was
added
10% Pd on carbon (2mg). Mixture was treated with hydrogen at 40 psi overnight.
Reaction mixture was filtered through a pad of Celite, and concentrated in
vacuo. The
residue was purified by HPLC to give the title compound (Smg, yield: 56%) as a
fluffy
white powder. ES-MS (M+H+): 513.1.
Example 20. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-
amidinophenyl)-2-fluoro-3-methylpropionamide. MS (M+H+): 455.1.
S02NH2
H
~N \ / \ /
O
H2N NH
Example 21. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-
amidinophenyl)- 3-isopropylpropionamide.
S02NH2
H
N \ / \ /
O
H2N NH
A. Preparation of 3-isopropylbenzenecarbonitrile.
To a mixture prepared by adding copper cyanide (940mg, 10.5mmol) to a
cooled solution of lithium bromide (1.82g 21mmo1) in tetrahydrofurn at -
25°C under
argon atmosphere was added a solution of O.SM 3-cyanophenyl zinc iodide (20m1,
l Ommol) in tetrahydrofuran. The reaction mixture was allowed to warm to 0
°C for 30
minutes then cooled down to -25 °C to which neat isobutyryl chloride
(1.06m1,
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l0.lmmol) was added all at once. The reaction was kept at -25 °C for 30
minutes then
quenched by adding 20m1 of a saturated solution of ammonium chloride. The
mixture
was extracted with 2x25m1 diethyl ether. The combined organic layers were
dried over
magnesium sulfate, filtered and concentrated in vacuo to an oil. The crude oil
was
flushed through a silica plug using 10% ethyl acetate in hexane to give 3-
isopropylbenzenecarbonitrile (1.25g, 72%) as a clear oil. H'NMR (CDCl3) : 2.2-
2.25
(d, 6H); 4.499-4.568 (m, H); 8.614-8.655 (m, 2H); 8.831-8.857 (m, H); 9.172-
9.238
(m, H). ES-MS (M+H+): 174.1.
B. Preparation of (2Z) methyl 3-(3-cyanophenyl)-3-isopropylacrylate
To a solution of bis(2,2,2-trifluoromethyl)(methoxy
carbonylmethyl)phosphonate (0.38m1, l.8mmo1) in 2.5m1 anhydrous
tetrahydrofuran
was added a solution of 18-Crown-6 (1.9g, 7.Smmo1) in 2.5m1 anhydrous
tetrahydofuran. The reaction mixturewas cooled to -78 °C under argon to
which was
added a solution of O.SM bis(trimethylsilyl)amide in toluene (3.6m1, l.8mmol).
The
reaction was stirred at -78 °C for 15 minutes to which was added a
solution of 3-
isopropylbenzenecarbonitrile in 2.5m1 anhydrous tetrahydoft~ran. The reaction
was
allowed to warm to room temperature and stirred for 48 hours. The reaction was
quenched by the addition of 20m1 of a saturated ammonium chloride solution
followed
by extraction with 2x25m1 diethyl ether. Combined organic layers were washed
with
2x25m1 water, 2x25m1 saturated brine solution, dried over magnesium sulfate,
filtered
and concentrated to give a 9:1 mixture of Z and E isomers (420mg, 120%) as a
clear oil
which was sufficiently pure to use without further purification. ES-MS (M+H+):
230.1.
C. Preparation of (2Z)-N-{4-[(2-tert-butylaminosulfonyl)phenyl]phenyl}-3-(3-
cyanophenyl)-3-isopropylacrylamide
To a solution of 2'-tert-butylaminosulfonyl-4-amino-[1,1']-biphenyl (139mg,
0.46mmo1) in 4m1 anhydrous dichloromethane was added a solution of 2M
trimethylaluminum in hexane (0.69m1, 1.38mmo1). Reaction was stirred at room
temperature for 20 minutes to which a solution of crude methyl (2Z)-3-(3-
cyanophenyl)-4-methylpent-2-enoate (lOSmg, 0.46mmo1) in 2m1 anhydrous
dichloromethane was added. Reaction was stirred at room temperature overnight.
Reaction was quenched with Sml 1N HCI after which an additional 20m1
dichloromethane was added. Organic was washed with 2x20m1 water, dried over
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magnesium sulfate, filtered and concentrated to give the title compound
(190mg, 82%)
as an off white foam which was sufficiently pure to be used without further
purification. ES-MS (M+H+): 501.2.
D. Preparation of (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-
amidinophenyl)-3-isopropylacrylamide
To a solution of crude (2Z)-N-{4-[(2-tert-butylaminosulfonyl)phenyl]phenyl)-
3-(3-cyanophenyl)-3-isopropylacrylamide (190mg, 0.379mmol) in lOml 1:1 ethyl
acetate:anhydrous methanol cooled to -78°C was bubbled HCl gas until
saturation was
achieved. Reaction was allowed to warm to room temperature and stirred
overnight.
The reaction was then concentrated in vacuo and dried under hi vacuum. The
dried
methyl imidate residue was dissolved in Sml anhydrous methanol to which
ammonium
acetate (115mg, l.Smmo1) was added and the reaction heated to reflux for 2
hours. The
reaction was concentrated and purified on a 2x25cm Vydac C,8 HPLC column to
the
title compound (75mg, 43%) as a fluffy white powder after lyophilization. ES-
MS
(M+H+): 463.2.
E. N-[4-(2-aminosulfonylphenyl)phenyl]-3-isopropyl-3-(3-amidinophenyl)-
propionamide.
To a solution of (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl{-3-(3-
amidinophenyl)-3-isopropylacrylamide (7mg) in 4m1 methanol was added 10% Pd on
carbon (2mg). Mixture was treated with hydrogen at 1 atmosphere under balloon
for
lhr. Reaction was filtered through a pad of Celite, concentrated and
lyophilized to give
the title compound (7mg, 100%) as a fluffy white powder. ES-MS (M+H+): 465.2.
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Example 22. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-2-hydroxyl-3-
methylidenyl-3-(3-amidinophenyl)-propionamide and N-[4-(2-
aminosulfonylphenyl)phenyl]-2,3-dihydroxyl-3-methyl-3-(3-amidinophenyl)-
propionamide.
HO S02NH2 H3C HO H S02NH2
H
I H N \ / \ / I ~ OH N \ / \ /
i p O
H2N NH H2N NH
A. tert-Butyl3-methyl-3-(3-cyanophenyl)-2,3-epoxypropanoate.
To a solution of 3-acetylbenzonitrile (1.45 g, 10 mmol) and tert-butyl
chloroacetate
(1.43 mL, 10 mmol) in tent-butanol (60 mL) at room temperature, potassium t-
butoxide
(1.45 g, 12.9 mmol) was added. The reaction mixture was stirred at room
temperature
overnight. Aqueous ammonium chloride was added to quench the reaction. Ethyl
acetate and water were added. The organic phase was separated, washed with
brine,
dried over Na2S04, concentrated in vacuo. The residue was purified by a silica
gel
column, first eluted with hexane, followed by 5% and 10% ethyl acetate in
hexane, to
give the title compound as a mixture of stereoisomers (1.49 g, yield: 58%). 'H
NMR
(CDC13) 7.76 - 7.40 (m, 4H), 3.60 (s, 1H, major isomer, 60%), 3.33 (s, 1H,
minor
isomer, 40%), 1.80 (s, 3H, minor isomer), 1.75 (s, 3H, major isomer), 1.55 (s,
9H,
minor isomer), 1.08 (s, 9H, major isomer).
B. 3-Methyl-3-(3-cyanophenyl)-2,3-epoxypropionic acid.
To a solution of tert-Butyl 3-methyl-3-(3-cyanophenyl)-2,3-epoxypropanoate
(0.24
g, 0.93 mmol) in methanol (S mL), 5 N NaOH (1.0 mL, 5.0 mmol) was added. The
solution was stirred at room temperature for 3 hrs. It was neutralized with 6N
HCl to
pH = 1-2. Ethyl acetate and water were added. The organic phase was separated,
washed with brine, dried over Na2S04, concentrated in vacuo to give an oil
(0.17 g,
yield: 90%). MS (M+H) 204.
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C. N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-methyl-3-(3-cyanophenyl)-
2,3-
epoxypropionamide.
To solution of 3-Methyl-3-(3-cyanophenyl)-2,3-epoxypropionic acid (168 mg,
0.828 mmol) and 4-(2-tert-butylaminosulfonylphenyl)aniline (252 mg, 0.828
mmol)
and triethylamine (0.23 mL, 1.66 mmol) in anhydrous DMF (4 mL), BOP (735 mg,
1.66 mmol) was added. The reaction mixture was stirred at room temperature
overnight. Ethyl acetate and water were added. The organic phase was
separated,
washed with sat. NaHC03, dried over Na2S04, concentrated in vacuo to give a
solid
(402 mg, yield: 99%). MS (M+Na) 512.1; (M+H-tBu) 434Ø
D. N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-methyl-3-(3-amidinophenyl)-
2,3-
epoxypropionamide.
To a solution of N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-methyl-3-(3-
cyanophenyl)-2,3-epoxypropionamide (223 mg, 0.456 mmol) in ethanol (6 mL),
hydroxylamine hydrochloride (64 mg, 0.92 mmol) was added, followed by addition
of
triethylamine (0.190 mL, 1.37 mmol). The mixture was heated at 60 C overnight.
The
solution was concentrated in vacuo. The residue was dissolved in acetic acid
(3 mL).
To the solution, acetic anhydride (0.217 mL, 2.30 mmol) was added. The
reaction
mixture was stirred at room temperature for 30 min., then concentrated in
vacuo and
dried on high vaccuum. The residue was dissolved in methanol (S mL). To the
solution,
Palladium on carbon (5%, 25 mg) was added. The mixture was hydrogenated under
balloon H2 overnight. The solution was filtered through a plug of celite. The
filtrate
was concentrated in vacuo. The residue was purified by HPLC to give the title
compound (73 mg, yield: 32%). MS (M+H) 507.2.
E. N-[4-(2-aminosulfonylphenyl)phenyl]-2-hydroxyl-3-methylidenyl-3-(3-
amidinophenyl)-propionamide and N-[4-(2-aminosulfonylphenyl)phenyl]-2,3-
dihydroxyl-3-methyl-3-(3-amidinophenyl)-propionamide.
The compound N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-methyl-3-
(3-amidinophenyl)-2,3-epoxypropionamide (20 mg, 0.040 mmol) was dissolved in
TFA (1 mL). The solution was allowed to stand at room temperature overnight,
then
concentrated in vacuo. The residue was purified by HPLC to give the product N-
[4-
(2-aminosulfonylphenyl)phenyl]-2-hydroxyl-3-methylidenyl-3-(3-amidinophenyl)-
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propionamide (10 mg) and product N-[4-(2-aminosulfonylphenyl)phenyl]-2,3-
dihydroxyl-3-methyl-3-(3-amidinophenyl)-propionamide (5 mg). For compound N-
[4-(2-aminosulfonylphenyl)phenyl]-2-hydroxyl-3-methylidenyl-3-(3-
amidinophenyl)-propionamide, MS (M+H) 451.1; 'H NMR (CD30D) 8.09 (d, 1H),
7.91 (d, 1H), 7.90 (s, 1H), 7.68 (d, 1H), 7.63 - 7.47 (m, SH), 7.36 (d, 2H),
7.30 (d,
1H), 5.67 (d, 2H), 5.18 (s, 1H). For compound N-[4-(2-
aminosulfonylphenyl)phenyl]-2,3-dihydroxyl-3-methyl-3-(3-amidinophenyl)-
propionamide, MS (M+H) 469.1; 'H NMR (CD30D) 8.09 (d, 1H), 7.93 (s, 1H),
7.92 (d, 1H), 7.70 - 7.40 (m, 6H), 7.36 (d, 2H), 7.30 (d, 1H), 4.26 (s, 1H),
1.73 (s,
3H).
Example 23. Preparation of N-[4-(2-aminosulfonylphenyl)phenyl]-2-hydroxyl-3-
methyl-3-chloro-3-(3-amidinophenyl)-propionamide and N-[4-(2-
aminosulfonylphenyl)phenyl)-2-hydroxyl-3-methyl-3-methoxy1-3-(3-
amidinophenyl)-propionamide.
Me0 OH SO2NH2
CI OH SO2NH2 HsC H _ _
H
~H\ H N \ / \ / I ~ O N \ / \ /
O '-'
H2N NH
H2N NH
The compound N-[4-(2-tert-butylaminosulfonylphenyl)phenyl]-3-methyl-3-
(3-cyanophenyl)-2,3-epoxypropionamide (130 mg, 0.266 mmol), from Part C of
Example 22, was dissolved in anhydrous methanol (4 mL). To the solution cooled
in
ice bath, HCl gas was bubbled through until saturation was reached. It was
stirred at
room temperature overnight, and then concentrated in vacuo. The residue was
dissolve in anhydrous methanol (3mL). To the solution, ammonium acetate (144
mg,
1.87 mmol) was added. The mixture was allowed to stand at room temperature
overnight, and then concentrated in vacuo. The residue was purified by HPLC to
give the product N-[4-(2-aminosulfonylphenyl)phenyl]-2-hydroxyl-3-methyl-3-
chloro-3-(3-amidinophenyl)-propionamide (35 mg) and product N-[4-(2-
aminosulfonylphenyl)phenyl]-2-hydroxyl-3-methyl-3-methoxyl-3-(3-
amidinophenyl)-propionamide (11 mg). For compound N-[4-(2-
aminosulfonylphenyl)phenyl]-2-hydroxyl-3-methyl-3-chloro-3-(3-amidinophenyl)-
propionamide, MS (M+H) 487.0 and 489.0 (chlorine pattern); 'H NMR (CD30D)
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8.07(d, 1H), 8.05 (d, 1H), 8.00 (s, 1H), 7.75 - 7.28 (m, 9H), 4.68 (s, 1H,
major
isomer, 65%), 4.60 (s, 1H, minor isomer, 35%), 2.21 (s, 3H, minor isomer),
2.18 (s,
3H, major isomer). For compound N-[4-(2-aminosulfonylphenyl)phenyl]-2-
hydroxyl-3-methyl-3-methoxyl-3-(3-amidinophenyl)-propionamide, MS (M+H)
483.1; 'H NMR (CD30D) 8.09(d, 1H), 7.88 (d, 1H), 7.80 (s, 1H), 7.77 - 7.29 (m,
9H), 4.30 (s, 1H, minor isomer, 40%), 4.27 (s, 1H, major isomer, 60%), 3.25
(s, 3H,
minor isomer), 3.23 (s, 3H, major isomer), 1.82 (s, 3H, minor isomer), 1.78
(s, 3H,
major isomer).
Example 24. Preparation of N-[4-(1-pyrrolidinylcarbonyl)phenyl]-3-(3-
amidinophenyl)-propionamide.
0
N ~ I NH2
N ~ ~ H
NH
The compound was prepared analogously to preparation of N-[4-(2-
aminosulfonylphenyl)phenyl]-3-(3-amidinophenyl)-propionamide in Example 1. MS
(M+H) 365.1.
Example 25. Preparation of N-{4-[(2-aminosulfonyl)phenyl]phenyl{-3-(3-
amidinophenyl)-2-methylpropionamide
S02NH2
H
O N \ / \ /
H2N NH
Part A. Ethyl (Z)-3-(3-cyanophenyl)-2-methyl-2-butenoate
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A solution of 1.00 g (3.0 mmol) of ethyl 2-[bis(2,2,2-
trifluoroethyl)phosphono]-propionate (Synth. Comm., 1991, 21, 2391) and 3.9 g
(5
eq) of 18-crown-6 in 25 mL of anhydrous THF was cooled with a dry ice-acetone
bath, and 7.0 mL of a 0.5 M solution of potassium bis(trimethylsilyl)amide in
toluene were added. The solution was stirred in the cold for 20 min, then a
solution
of 400 mg (3.05 mmol) of 3-cyanobenzaldehyde in 10 of anhydrous THF was added
dropwise over a few minutes. The reaction was stirred in the cold for 1 hr,
then
allowed to warm to room temperature over 3 hr, quenched by the addition of 10
mL
of saturated aqueous ammonium chloride, and extracted with 2 x 50 mL of ether.
I0 The organic layer was washed with 50 mL of water, followed by 50 mL of
saturated
NaCI, then dried over MgS04. Filtration and concentration gave 1 g of a light
yellow oil, which was washed through a plug of silica gel with 200 mL of
CHzCIz.
Concentration then gave 586 mg (97%) of the desired product as a light yellow
oil,
which was >90% the desired (Z)-isomer by'H NMR: 'H NMR (CDCI3) 8 2.10 (s,
3H), 3.64 (s, 3H), 6.68 (s, 1H), 7.1-7.3 (m, 1H), 7.35-7.55 (m, 3H).
Part B. (2Z)-N-[4-(2{[(N-l,l-dimethylethyl)amino]sulfonyl}phenyl)phenyl]-3-(3-
cyanophenyl)-2-methylacrylamide
To a solution of 103 mg (0.34 mmol) of 4'-amino-N-(1,1-dimethylethyl)-
[1,1'-biphenyl]-2-sulfonamide in 5 mL of anhydrous CHZCIZwas added 0.5 mL of a
2.0 M solution of trimethylaluminum in hexanes, and the solution was stirred
at
room temperature for 30 minutes. A solution of 103 mg of ethyl (Z)-3-(3-
cyanophenyl)-2-methyl-2-butenoate in 5 mL of anhydrous CHzCIz was then added
dropwise over a few minutes, and the reaction was stirred at room temperature
overnight. The reaction was then carefully quenched by the addition of 10 mL
of
1N HCI, and the reaction mixture was then partitioned between 100 mL of CHZC12
and 50 mL of H20. The organic layer was dried over MgS04, filtered and
concentrated to give a solid residue, which was subj ected to flash column
chromatography on silica gel using 10% EtOAc in hexanes to give 104 mg (65%)
of
the desired product as a white solid: 'H NMR (CDC13) 8 1.00 (s, 9H), 2.22 (s,
3H),
3.60 (s, 1H), 6.59 (s, 1H), 7.15-7.6 (m, 12H), 8.14 (d, J = 7.6 Hz, I H).
Part C. (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl{-3-(3-amidinophenyl)-2-
methylacrylamide
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A suspension of 50 mg of the above nitrite in 10 mL of anhydrous methanol
was cooled with an ice-water bath, and HCl gas was bubbled into the solution
at a
moderate rate for 10 min. The reaction was then closed with a rubber septum
and
stirred at room temperature overnight. The reaction was concentrated to give a
semisolid residue, which was taken up in 5 mL of anhydrous methanol, and 41 mg
of vacuum dried ammonium acetate were added. The solution was heated at gentle
reflux for 1.5 hr, then concentrated to give a white solid. Preparative HPLC
(gradient elution with water: acetonitrile each containing 0.1 % TFA on C 18)
then
afforded 44 mg of the desired product as a white solid: 'H NMR (DMSO-d6) 8
2.17
(s, 3H), 6.59 (s, 1H), 7.25-7.35 (m, SH), 7.5-7.65 (m, 6H), 7.71 (d, J = 5.6
Hz, 1H),
8.01 (d, J = 7.6 Hz, 1H), 8.95 (s, 2H), 9.31 (s, 2H). 10.25 (s, 1H). MS 435.1.
Part D. N- f 4-[(2-aminosulfonyl)phenyl]phenyl}-3-(3-amidinophenyl)-2-
methylpropanamide
A solution of 14 mg of the alkene from Part C and 5 drops of triethyamine in
5 mL of methanol, together with 10 mg of 10% Pd/C was placed under a balloon
of
hydrogen and stirred overnight. The reaction was filtered and concentrated,
and the
residue was subjected to preparative HPLC (gradient elution with
water:acetonitrile
each containing 0.1 % TFA on C 18) to give 7 mg of the desired product as a
white
solid:'H NMR (CD30D) 8 1.24 (d, J = 6.4 Hz, 3H), 2.82 (d, J = 9.2 Hz, 2H),
3.02
(m, 1H), 7.2-7.65 (m, 11H), 8.02 (d, J = 7.6 Hz, 1H). MS (M+H) 437.1.
Example 26. N- f 4-[(2-aminosulfonyl)phenyl]phenyl}-3-(1-aminoisoquinolin-7-
yl)-
propionamide
S02NH2 O NH2
/ \ / \ NH i I ~N
Part A. 7-Bromoisoquinoline
This compound was prepared as a 60:40 mixture with 5-bromoisoquinoline as in
J.
Am. Chem. Soc., 1939, 61, 183.
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Part B. 7-Bromoisoquinoline N-oxide hydrochloride
This compound was prepared by a procedure analogous to that for 6-
bromoisoquinoline N-oxide hydrochloride as in PCT WO 98/47876. A solution of
7.8 g (37.5 mmol) of a 60:40 mixture of 7-bromo and 5-bromoisoquinoline in 125
mL of CHZC12 was treated portionwise with 9.7 g 039.4 mmol) of 3-
chloroperoxybenzoic acid (~70% purity). The solution, which was initially
homogeneous, deposited a voluminous precipitate over 1 hr. Then 100 mL of
methanol were added, and the reaction was concentrated to a volume of about
100
mL. Gaseous HCl was then bubbled through the solution for about 10 min, during
which time the solution became warm and all of the precipitate dissolved. A
few
minutes later, another voluminous precipitate began to form. To this solution
was
added 100 mL of ether, and the mixture was stirred in an ice-water bath for 20
minutes. The resulting product was isolated by filtration, washed thoroughly
with
ether, and air-dried to give 8.07 g (83%) of the desired compound as a white
solid,
which was still a 60:40 mixture of the 7- and 5-bromo isomers.
Part C. 7-Bromo-1-chloroisoquinoline
This compound was prepared by a procedure analogous to that for 6-bromo-
1-chloroisoquinoline as in PCT WO 98/47876. A solution of 8.07 g (31 mmol) of
the mixture from Part B was taken up in 50 mL of POC13, and the mixture was
heated at 90 °C for 2 hr. The reaction mixture was concentrated to
remove most of
the POC13, and the residue was taken up in 100 mL of CHZC12. The solution was
carefully basified to pH 10 by the slow addition of 1N NaOH, and the organic
layer
was washed with 100 mL of H20, 100 mL of sat. NaCI, and dried over MgS04.
Filtration and concentration gave a light yellow solid, which was subjected to
flash
column chromatography on silica gel first with 5% and then with 10% EtOAc in
hexanes. A total of 3.62 g (48%) of the desired 7-bromo-1-chloroisoquinoline
was
isolated from this chromatography free of the 5-bromo isomer.
Part D. 7-Bromo-1-phenoxyisoquinoline
A solution of 3.60 g (14.8 mmol) of 7-bromo-1-chloroisoquinoline and 1.5 g
of solid KOH in 11.2 g of phenol was heated at 140 °C for 2 hr. The
reaction was
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cooled to room temperature, then partitioned between 100 mL of CHZCIz and 100
mL of 3N NaOH. The organic layer was washed with another 2 x 100 mL of 3N
NaOH, then with 100 mL of H,O, and dried over MgS04. Filtration and
concentration gave a yellow oil, which was subj ected to flash column
chromatography on silica gel 30% CHzCIz in hexanes, giving 3.42 g (77%) of the
desired product as a light yellow solid.
Part E. 1-Amino-7-bromoisoquinoline
A mixture of 3.40 g (11.3 mmol) of 1-amino-7-bromoisoquinoline and 7.65 g
of ammonium acetate was heated at 150 °C for 1 S hr. The reaction was
cooled, and
the residue was partitioned between 200 mL of EtOAc and 200 mL of 3N NaOH.
The organic layer was extracted with 2 x 100 mL of 2N HCI, and the combined
aqueous extracts were basified to pH 10 using 50% NaOH. This solution was
extracted with 2 x 100 mL of EtOAc, and the organics were then washed with 100
mL of sat. NaCI and dried over MgS04. Filtration and concentration gave 1.68 g
(66%) of the desired amino compound as a yellow solid.
Part F. 1-[Bis(t-butoxycarbonyl)amino]-7-bromoisoquinoline
A solution of 740 mg (3.32 mmol) of 1-amino-7-bromoisoquinoline in 50
mL of acetonitrile was treated with 1.4 mL of N,N-diiospropylethylamine and
100
mg of 4-(N,N-dimethylamino)pyridine, followed by 3.0 g (4.1 eq) of di-t-
butyldicarbonate, and the reaction was stirred at 40 °C for 1 hr. By
HPLC analysis,
there was still some starting amino compound that remained , so another 1.0 g
of di-
t-butyldicarbonate were added, and the reaction was stirred at 40 °C
for another 30
min. The reaction mixture was concentrated to give a dark oil, which was
subjected
to flash column chromatography on silica gel with 20% EtOAc in hexanes to give
736 mg of the desired product as a light yellow solid. Also isolated were 156
mg of
product as a somewhat less pure light yellow solid, making the total yield
64%.
Part G. 1-[Bis(t-butoxycarbonyl)amino]isoquinoline-7-carboxaldehyde
A solution of 400 mg (0.95 mmol) of 1-[bis(t-butoxycarbonyl)amino]-7-
bromoisoquinoline in 50 mL of anhydrous THF was cooled with a liquid
nitrogen/methanol slush bath (-98 °C), and 0.55 mL of a 2.43 M solution
of n-BuLi
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in hexanes (1.3 eq) was added dropwise over 1 min. The solution was stirred in
the
cold for 5 min, then a solution of 5 mL of anhydrous DMF in 10 mL of anhydrous
THF was added rapidly. The solution was allowed to warm to about 0
°C, then
poured into 50 mL of 0.5 N HCI, and 50 mL of EtOAc were added. The aqueous
layer was brought to pH 6 with 1N NaOH, 25 mL of sat. NaCI were added, and the
layers were shaken and separated. The organic layer was dried over Mg S04,
filtered, and concentrated to give an oily residue. This residue was subjected
to
flash column chromatography on silica gel with 20% EtOAc in hexanes to give
190
mg (54%) of the desired aldehyde as a yellow semisolid.
Part H. (2Z)-3-{[1-bis(t-butoxycarbonyl)amino]isoquinolin-7-yl}acrylic acid, 2-
(trimethylsilyl)ethyl ester
A solution of 117 mg (0.29 mmol) of [bis(2,2,2-
trifluoroethoxy)phosphinyl]acetic acid, 2-(trimethylsilyl)ethyl ester (J. Org.
Chem.,
1991, 56, 4204) and 400 mg of 18-crown-6 in 25 mL of anhydrous THF was cooled
with a dry ice-acetone bath under Ar, and 0.75 mL of a 0.5 M solution of
potassium
bis(trimethylsilyl)amide in toluene were added dropwise over 2 min. The
reaction
was stirred in the cold for 15 min, then a solution of 100 mg (0.27 mmol) of 1-
[bis(t-
butoxycarbonyl)amino]isoquinoline-7-carboxaldehyde in 25 mL of anhydrous THF
was added dropwise over 10 min. The reaction was then allowed to warm to room
temperature overnight, then partitioned between 100 mL of CHZC12 and 50 mL of
HZO. The organics were washed with aqueous NaCI, and dried over MgS04.
Filtration and concentration gave an oily residue, which was subj ected to
flash
column chromatography on silica gel with 25% EtOAc in hexanes to give 33 mg of
the desired product as a clear, colorless oil.
Part L (2Z)-N-[4-(2{[(N-1,1-dimethylethyl)amino]sulfonyl}phenyl)phenyl]-3-{[1-
bis(t-butoxycarbonyl)amino]isoquinolin-7-yl} acrylamide
A solution of 63 mg (0.12 mmol) of (2Z)-3-{[1-bis(t-
butoxycarbonyl)amino]isoquinolin-7-yl}acrylic acid, 2-(trimethylsilyl)ethyl
ester in
1 mL of DMF was treated at room temperature with 150 ~L of 1.0 M
tetrabutylammonium fluoride in THF overnight. The reaction mixture was
directly
subjected to preparative HPLC (gradient elution with water:acetonitrile each
containing 0.1 % TFA on C 18) to give, after lyophilization, 21 mg of the
desired acid
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as a white solid. A solution of this acid and 18 mg of 4'-amino-N-(1,1-
dimethylethyl)-[1,1'-biphenyl]-2-sulfonamide in 2 mL of anhydrous DMF,
together
with 40 ~,L of N,N-diisopropylethylamine, was treated at room temperature with
25
mg (1.3 eq) of HATU, and the reaction was stirred at room temperature for 1
hr.
The reaction mixture was dissolved in 100 mL of CHZC12, washed with 2 x 25 mL
of
sat. NaHC03, and dried over MgS04. Filtration and concentration gave 53 mg of
the
desired product as a yellow oily residue, which was used in the next reaction
without
further purification.
Part J. (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(1-aminoisoquinolin-7-yl)-
acrylamide
A solution of the yellow oil from Part I in 2 mL of TFA was stirred first in
an
ice-water bath, and then at room temperature overnight. The reaction mixture
was
concentrated and directly subjected to preparative HPLC (gradient elution with
water:acetonitrile each containing 0.1 % TFA on C 18) to give 10 mg of the
desired
product as an off white solid.
Part K. N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(1-aminoisoquinolin-7-yl)-
propionamide
A solution of 11 mg of (2Z)-N-{4-[(2-aminosulfonyl)phenyl]phenyl}-3-(1-
aminoisoquinolin-7-yl)-acrylamide in 10 mL of methanol, together with a few mg
of
10% Pd/C was placed under a balloon of hydrogen gas for 1 hr. At this time,
the
reaction was complete by HPLC (the retention time was the same as the starting
material, but the UV spectrum had changed significantly). The reaction mixture
was
filtered and concentrated to give the crude product as an oil. A small amount
of
water was added, and the mixture was lyophilized to give the desired compound
as a
white solid: 'H NMR (CD30D) 8 2.80 (t, J = 7.2 Hz, 2H), 3.23 (t, J = 7.6 Hz,
2H),
7.16 (d, J = 6.8 Hz, 1H), 7.27 (d, J = 7.2 Hz, 1H), 7.31 (d, J = 8.0 Hz, 2H),
7.45-7.5
(m, 4H), 7.56, (app t, J = 7.2 Hz, 1 H), 7.84 (d, J = 8.4 Hz, 1 H), 7.90 (app
d, J = 8.0
Hz, 1H), 8.05 (app d, J = 8.0 Hz, 1H). 8.26 (s, 1H). . MS (M+H) 447.1.
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Example 27. Preparation of 3-(1-amino(7-isoquinolyl))-3-chloro-2-hydroxy-N-[4-
(2-
sulfamoylphenyl)phenyl]butanamide
OH
H
N SO NH
O I ~ 2 z
N~ NH2 I
A. Preparation of 7-isoquinolyl (trifluoromethyl)sulfonate
To a solution of 7-hydroxyisoquiniline (S.SOg, 37.9mmo1) in 100m1 CHzCl2
was added triethylamine (B.OOmI, 57.4mmo1) dropwise. Then DMAP (0.275g,
2.24mmol) was added. The reaction was stirred for Smin before being cooled to
0°C
for l5min. Tf20 (9.5m1, 56.Smmo1) was added and the reaction was allowed to
warm to room temperature overnight. Water and DCM were added and separated.
The aqueous layer was extracted two more times with DCM. The combined organic
layers were dried over MgS04, filtered and concentrated. The residue was
purified
by silica gel, first eluded with 20/80 EtOAc/hexanes, then 25/85 EtOAc/hexanes
and
finally with 30/70 EtOAc/hexanes to give the title compound (5.12g). MS 278 (M
+
H)
B. Preparation of isoquinoline-7-carbonitrile
To a solution of Pd2(dba)3 (0.958g, 0.926mmo1) and dppf (1.80g, 3.25mmo1)
in DMF (18m1) was added 7-isoquinolyl (trifluoromethyl)sulfonate (5.12g,
l8.Smmol). Heat reaction mixture to 70°C and add Zn(CN)2 (1.30g,
ll.lOmmol) in
three portions (~0.43g each) every l5min. The reaction was allowed to stir for
3
hours before being quenched with water and extracted with EtOAc three times.
The
combined organic layers were dried over MgS04, filtered and concentrated. The
residue was purified by silica gel, first eluded with 20/80 EtOAc/hexanes,
25/85
EtOAc/hexanes, 30/70 EtOAc/hexanes, and 35/75 EtOAc/hexanes to give the title
compound (1.75g). MS 155 (M + H)
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C. Preparation of 1-(7-isoquinolyl)ethan-1-one
To a solution of isoquinoline-7-carbonitrile (0.496g, 3.22mmo1) in 20m1
EtzO at 0°C was added MeMgBr (13.8m1, 19.3mmo1) dropwise. The
reaction was
allowed to stir at room temperature for ~3hrs before being quenched with
saturated
NHQCI. The mixture was neutralized to pH=8 with SM NaOH, then extracted twice
with Et20. The combined organic layers were dried over MgS04, filtered and
concentrated. The residue was purified by silica gel, first eluded with 20/80
EtOAc/hexanes, 25/85 EtOAc/hexanes, 30/70 EtOAc/hexanes, and 35/75
EtOAclhexanes to give the title compound (0.389g). MS 172 (M + H)
D. Preparation of tert-butyl 3-(7-isoquinolyl)-3-methyloxirane-2-carboxylate
To a solution of 1-(7-isoquinolyl)ethan-1-one (0.109g, 0.637mmol) in
tBuOH (7m1) was added t-butyl chloroacetate (0.182mL, 1.27mmol) then K'Bu0
(0.147g, 1.31mmol). The reaction was heated to 40°C for 2 hr. Reaction
not
complete therefore more t-butyl chloroacetate (0.091u1, 0.53mmol) was added.
The
reaction was heated for a few more hours before being quenched with AcOH
(1m1).
Water and EtOAc were added. Aqueous layer was extracted twice more with
EtOAc. The combined organic layers were dried over MgS04, filtered and
concentrated to a residue (0.206g). MS 286 (M+H)
E. Preparation of 3-(7-isoquinolyl)-3-methyloxirane-2-carboxylic acid
To a solution of tert-butyl 3-(7-isoquinolyl)-3-methyloxirane-2-carboxylate
in MeOH (5m1) was added SM NaOH (0.75m1, 3.75mmo1). The reaction was
allowed to stir for 2hrs before being concentrated and purified by Preparatory
HPLC
to yield the title compound (0.402g). MS 230 (M+H)
F. Preparation ofN-[4-(2-{[(tent-butyl)amino]sulfonyl}phenyl)phenyl](3-(7-
isoquinolyl)-3-methyloxiran-2-yl)carboxamide
To a solution of N-[4-(2- f [(tert-butyl)amino]sulfonyl}phenyl)phenyl](3-(7-
isoquinolyl)-3-methyloxiran-2-yl)carboxamide (0.40g, 1.76mmo1), {[2-(4-
aminophenyl)phenyl]sulfonyl}(tent-butyl)amine (0.53g, 1.75mmol), and BOP in
DMF(lOml) was added TEA (0.981m1, 7.04mmo1) dropwise. The reaction was
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stirred at room temperature for 3hrs. Water and EtOAc were added. The aqueous
layer was extracted twice with EtOAc. The combined organic layers were dried
over
MgS04, filtered and concentrated. The residue was purified by silica gel to
yield the
title compound (0.2378, 0.46mmol). MS 516 (M+H)
G. Preparation of [3-(1-amino(7-isoquinolyl))-3-methyloxiran-2-yl]-N-[4-(2-
{[(tert-butyl)amino]sulfonyl}phenyl)phenyl]carboxamide
To a solution of N-[4-(2-{[(tert-butyl)amino]sulfonyl)phenyl)phenyl](3-(7-
isoquinolyl)-3-methyloxiran-2-yl)carboxamide in acetone (8m1) was added MCPBA
(0.0918, max 77%, 0.29mmo1). Approximately lhr later more MCPBA (0.0348,
1.06mmo1) was added. Approximately lhr later the reaction mixture was
concentrated and the residue was dissolved in sat. NaHC03 and EtOAc. The
aqueous layer was extracted once more with EtOAc. The combined organic layers
were dried over MgS04, filtered and concentrated. The residue was dissolved in
pyridine (7m1) and tosyl chloride (0.0668, 0.346mmo1). The reaction was
allowed to
stir overnight before being concentrated. The residue was dissolved in ~Sml
ethanolamine and allowed to stir for a few hours. Water was added to the
reaction
and the solid was collected MS 531 (M+H)
H. Preparation of 3-(1-amino(7-isoquinolyl))-3-chloro-2-hydroxy-N-[4-
(2 sulfamoylphenyl)phenyl]butanamide
To a solution of [3-(1-amino(7-isoquinolyl))-3-methyloxiran-2-yl]-N-[4-(2-
{[(tert-butyl)amino]sulfonyl}phenyl)phenyl]carboxamide (0.0608, 1.13mmo1) in
MeOH (5m1) was bubbled HCl gas to saturation. The reaction was capped with a
septum and allowed to stir overnight. Preparatory HPLC purification usingl0/90
acetonitrile (containing 0.1 % TFA) /HQ water (containing 0.1 %TFA) to 70/30
acetonitrile (containing 0.1 % TFA) /HQ water (containing 0.1 % TFA) over
60minutes gave the title compound. MS 511 (M+H)
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Example 28. Preparation of 3-(2-(2-pyridyl)-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl}
ethyl)benzenecarboxamidine
A. Preparation of ethyl (2Z)-3-(3-cyanophenyl)-2-(2-pyridyl)prop-2-enoate
A solution of 3-cyanobenzaldehyde (0.797g, 6.07mmo1), ethyl 2-
pyridylacetate (0.670m1, 6.07mmol) and ammonium acetate (0.566g, 7.39mmo1) in
acetic acid (3m1) was refluxed overnight. The reaction was cooled to room
temperature and neutralized to pH=7 with 5M NaOH/H20. EtOAc was added and
the aqueous layer was washed twice with EtOAc. The combined organic layers
were
dried over MgS04, filtered and concentrated. The residue was purified by
silica gel,
first eluded with 10/90 EtOAc/hexanes, 20/80 EtOAc/hexanes, 30/70
EtOAc/hexanes, 35/65 EtOAc/hexanes, 40/60 EtOAc/hexanes, 45/55
EtOAc/hexanes and finally with 50/50 EtOAc/hexanes to give the title compound
(0.407g). MS 279 (M + H)
B. Preparation of (2Z)-N-[4-(2- f [(tert-butyl)amino]sulfonyl}phenyl)phenyl]-3-
(3-cyanophenyl)-2-(2-pyridyl)prop-2-enamide
To a mixture of ethyl (2Z)-3-(3-cyanophenyl)-2-(2-pyridyl)prop-2-enoate
(0.210g, 0.758mmol) and f [2-(4-aminophenyl)phenyl]sulfonyl}(tert-butyl)amine
(0.235g, 0.773mmo1) in CHzCl2 (5m1) was added AlMe3 (2m1 of 2M in hexane,
4mmo1). The mixture was allowed to stir overnight before quenching with 6M
HCI.
The aqueous layer was extracted twice more with DCM. The combined organic
layers were dried over MgS04, filtered and concentrated. The residue was
purified
by silica gel, first elude with 20/80 EtOAc/hexanes, 30/70 EtOAc/hexanes,
40/60
EtOAc/hexanes, 45/55 EtOAclhexanes, 50/50 EtOAc/hexanes, 55/45
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EtOAc/hexanes, 60/40 EtOAc/hexanes, 70/30 EtOAc/hexanes, then with 80/20
EtOAc/hexanes to give the title compound (0.040g). MS 536 (M + H)
C. Preparation of 3-((1Z)-2-(2-pyridyl)-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl}vinyl)benzenecarboxamidine
Through a mixture of (2Z)-N-[4-(2-{[(tert-
butyl)amino]sulfonyl}phenyl)phenyl]-3-(3-cyanophenyl)-2-(2-pyridyl)prop-2-
enamide (0.226g, 0.421mmo1) in MeOH (7m1) was bubbled HCl gas to saturation.
The reaction was capped with a septum and allowed to run overnight. The
reaction
was concentrated and the residue was dissolved in MeOH (7m1) to which was
added
NH40Ac (0.194g, 2.56mmo1). The mixture was refluxed for 4hrs before being
concentrated. The residue was purified by preparatory HPLC 10/90 acetonitrile
(containing 0.1 % TFA) /HQ water (containing 0.1 % TFA) to 90/10 acetonitrile
1 S (containing 0.1 % TFA) /HQ water (containing 0.1 % TFA) over 80minutes to
yield
the title compound (0.212g). MS 498 (M+H)
D. Preparation of 3-(2-(2-pyridyl)-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl} ethyl)benzenecarboxamidine
A mixture of 3-((1Z)-2-(2-pyridyl)-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl}vinyl) (0.043g, 0.087mmo1) Pd/C (Smg) in
MeOH (4m1) with a HZ balloon was allowed to stir overnight. The reaction was
filtered over celite and concentrated. Preparatory purification yielded the
target
compound. MS 500 (M+H)
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Example 29. Preparation of 3-(2-(3-pyridyl)-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl}
ethyl)b enzenecarboxamidine
'1
\ \ N
O"NH
N NHz
~S\~ NHz
O
A. Preparation of ethyl (2Z)-3-(3-cyanophenyl)-2-(3-pyridyl)prop-2-enoate
A solution of 3-cyanobenzaldehyde (0.795g, 6.OSmmo1), ethyl 3
pyridylacetate (0.666m1, 6.07mmol) and ammonium acetate (0.569g, 7.387mmo1) in
acetic acid (3m1) was refluxed overnight. The reaction was cooled to room
temperature and neutralized to pH=7 with SM NaOH/H20. EtOAc was added and
the aqueous layer was washed twice with EtOAc. The combined organic layers
were
dried over MgS04, filtered and concentrated. The residue was purified by
silica gel,
first eluded with 10/90 EtOAc/hexanes, 20/80 EtOAc/hexanes, 30/70
EtOAc/hexanes, 35/65 EtOAc/hexanes, 40/60 EtOAc/hexanes, 45/55
EtOAc/hexanes and finally with 50/50 EtOAc/hexanes to give the title compound
(0.365g). MS 279 (M + H)
B. Preparation of (2Z)-N-[4-(2-{[(tert-butyl)amino]sulfonyl}phenyl)phenyl]-3-
(3-
cyanophenyl)-2-(3-pyridyl)prop-2-enamide
To a mixture of ethyl (2Z)-3-(3-cyanophenyl)-2-(3-pyridyl)prop-2-enoate
(0.208g, 0.749mmo1) and {[2-(4-aminophenyl)phenyl]sulfonyl}(tert-butyl)amine
(0.228g, 0.750mmol) in CHzCl2 (5m1) was added AlMe3 (1.2m1 of 2M in hexane,
2.4mmo1). The mixture was allowed to stir overnight before quenching with 6M
HCI. The aqueous layer was extracted twice with DCM. The combined organic
layers were dried over MgS04, filtered and concentrated. The residue was
purified
by silica gel, first elude with 30/70 EtOAc/hexanes, 40/60 EtOAc/hexanes,
50/50
EtOAc/hexanes, 60/40 EtOAc/hexanes to give the title compound (0.157g). MS 536
(M+H)
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C. Preparation of 3-((1Z)-2-(3-pyridyl)-2-~N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl} vinyl)benzenecarboxamidine
Through a mixture of (2Z)-N-[4-(2- f [(tert-
butyl)amino]sulfonyl}phenyl)phenyl]-3-(3-cyanophenyl)-2-(3-pyridyl)prop-2-
enamide (0.157g, 0.293mmo1) in MeOH (7m1) was bubbled HCl gas to saturation.
The reaction was capped with a septum and allowed to run overnight. The
reaction
was concentrated and the residue was dissolved in MeOH (7m1) to which was
added
NH40Ac (0.138g, 1.79mmol). The mixture was refluxed for 4hrs before being
concentrated. The residue was purified by preparatory HPLC 5/95 acetonitrile
(containing 0.1 % TFA) /HQ water (containing 0.1 % TFA) to 95/5 acetonitrile
(containing 0.1 % TFA) /HQ water (containing 0.1 % TFA)water over 90minutes to
yield the title compound (0.237g). MS 498 (M+H)
D. Preparation of 3-(2-(3-pyridyl)-2-{N-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl} ethyl)benzenecarboxamidine
A mixture of 3-((1Z)-2-(3-pyridyl)-2- fN-[4-(2-
sulfamoylphenyl)phenyl]carbamoyl}vinyl)benzenecarboxamidine (0.053g,
0.087mmo1) Pd/C (Smg) in MeOH (5m1) with a HZ balloon was allowed to stir
overnight. The reaction was filtered over celite and concentrated. The residue
was
purified by preparatory HPLC 5/95 acetonitrile (containing 0.1 % TFA) /HQ
water
(containing 0.1 % TFA) water to 95/5 acetonitrile (containing 0.1 % TFA) /HQ
water
(containing 0.1 % TFA) water over 60minutes to yield the title compound
(0.016g).
MS 500 (M+H)
Example 30. Preparation of [S-(2-aminosulfonylphenyl)indolin-1-yl] 3-(3-
amidinophenyl)-propionyl amide.
NH2
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The compound was prepared analogously to preparation of N-[4-(2-
aminosulfonylphenyl)phenyl)-3-(3-amidinophenyl)-propionamide in Example 1. MS
(M+H) 449.1.
Example 31. Preparation of 3-(1-aminoisoquinol-7y1)-2-phenyl-N-[4-(2-
sulfamoylphenyl)phenyl]propionamide
O
",O
H2N-S
Ph
NH
I I
O
N~NH2
A. Preparation of ethyl 2-phenylprop-2-enoate
A mixture of ethyl 2-phenylacetate (15.9 mL, 100 mmol), paraformaldehyde
(4.5 g, 150 mmol), K2C03 (22.11 g, 100 mmol) and Bu4NI (0.7388 g, 2 mmol) in
toluene was heated to 80-90 C for 40 h. Standard workup (saturated NaCI
solution
washing) yielded 14.78 g (84%) of the titled compound. MS 177 (M+1).
B. Preparation of ethyl 3-(7-isoquinolyl)-2-phenylprop-2-enoate
traps-Di(~-acetato)bis[o-(di-o-tolylphosphino)benzyl]dipalladium(II)
(0.1654 g, 0.18 mmol) was added to a solution of 7-isoquinolyl
(trifluoromethyl)sulfonate (1.70 g, 7 mmol), ethyl 2-phenylprop-2-enoate (2.26
g,
7mmo1), and triethyl amine (1.95 mL, 14 mmol) in DMF (50 mL). After stirring
for
16 h at 120 C, Pd catalyst (0.1654 g, 0.18 mmol) was added again, and the
suspension was stirred for 16 h at 120 C. After standard workup (filtration
and
washing with saturated NaCI solution), the crude brown oil was purified by
Prep
HPLC to yield the titled compound 1.06 g (50%). MS 304 (M + 1).
C. Preparation ofN-[4-(2-([(tert-butyl)amino]sulfonyl}phenyl)phenyl]-3-(7-
isoquinolyl)-2-phenylpropanamide
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To a solution of compound ethyl 3-(7-isoquinolyl)-2-phenylprop-2-enoate
(172.5 mg, 0.57 mmol) in anhydrous MeOH (10 mL) was added 10% Pd/C (600
mg). The solution was then stirred under hydrogen ballon (1 atm) for 6 h. The
hydrogenated product of ethyl 3-(7-isoquinolyl)-2-phenylpropanoate was
collected
by the removal of the solvent in vacuo, and carried over to the next step
directly.
To a solution of 4-(2-tert-butylaminosulfonylphenyl)aniline (171.9 mg, 0.57
mmol) in CH2Cl2 (10 mL) at room temperature, trimethylaluminum (0.848 mL, 2.0
M in hexane, 1.7 mmol) was added dropwise. After the solution was stirred for
30
min at room temperature, compound ethyl 3-(7-isoquinolyl)-2-phenylpropanoate
from the last reaction was added. The mixture was stirred at room temperature
for 1
days. The solution was neutralized with 1N HCl (10 mL) to pH = 1-2. Water and
CH2C12 were added, and organic phase was separated, dried over Na2S04,
concentrated in vacuo to give a yellowish soild, which was further purified by
Prep
HPLC to yield the titled compound 198.9 mg (yield: 62%). MS 564 (M+1).
D. Preparation of 3-(1-amino(7-isoquinolyl))-2-phenyl-N-[4-(2-
sulfamoylphenyl)phenyl]propanamide
To a solution of N-[4-(2- f [(tert-butyl)amino]sulfonyl}phenyl)phenyl]-3-(7-
isoquinolyl)-2-phenylpropanamide (31 mg, 0.06 mmol) in acetone (5 mL) at 23 C
was added mCPBA (20 mg, 0.08 mmol). The mixture was stirred at this
temperature for 18 h. The solvent was removed in vacuo. The residue was
partitioned between EtOAc and saturated NaHC03 solution. The organic layers
were collected, dried over Na2S04 and concentrated in vacuo.
The crude product was dissolved in dry pyridine (5 mL) and TsCI (15.7 mg,
0.08 mmol) was added. The mixture was stirred at 23 C for 5 minutes. The
solvent
of pyridine was removed in vacuo. The residue was dissolved in ethanolamine (5
mL). The mixture was stirred at 23 C for 3 h before pouring into saturated
NaCI
solution for partition. The organic layers were collected and concentrated in
vacuo
to afford a yellow residue.
The residue was dissolved in TFA (5 mL). The mixture was stirred at 23 C
for 18 h. The solvent was removed in vacuo and the crude product was further
purified by Prep HPLC to yield the titled compound 10 mg (32%). MS 523 (M+1).
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BIOLOGICAL ACTIVITY EXAMPLES
Evaluation of the compounds of this invention is guided by in vitro protease
activity assays (see below) and in vivo studies to evaluate antithrombotic
efficacy,
and effects on hemostasis and hematological parameters.
The compounds of the present invention are dissolved in buffer to give
solutions containing concentrations such that assay concentrations range from
0 to
100 p,M. In the assays for thrombin, prothrombinase and factor Xa, a synthetic
chromogenic substrate is added to a solution containing test compound and the
enzyme of interest and the residual catalytic activity of that enzyme is
determined
spectrophotometrically. The ICSp of a compound is determined from the
substrate
turnover. The IC50 is the concentration of test compound giving SO% inhibition
of
the substrate turnover. The compounds of the present invention desirably have
an
IC50 of less than 500 nM in the factor Xa assay, preferably less than 200 nM,
and
more preferred compounds have an ICSO of about 100 nM or less in the factor Xa
assay. The compounds of the present invention desirably have an IC50 of less
than
4.0 pM in the prothrombinase assay, preferably less than 200 nM, and more
preferred compounds have an ICSO of about 10 nM or less in the prothrombinase
assay. The compounds of the present invention desirably have an ICSp of
greater
than 1.0 pM in the thrombin assay, preferably greater than 10.0 ~M, and more
preferred compounds have an ICSO of greater than 100.0 pM in the thrombin
assay.
Amidolvtic Assays for determining protease inhibition activity
The factor Xa and thrombin assays are performed at room temperature, in
0.02 M Tris~HCl buffer, pH 7.5, containing 0.15 M NaCI. The rates of
hydrolysis of
the para-nitroanilide substrate S-2765 (Chromogenix) for factor Xa, and the
substrate Chromozym TH (Boehringer Mannheim) for thrombin following
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preincubation of the enzyme with inhibitor for 5 minutes at room temperature,
and
were determined using the Softmax 96-well plate reader (Molecular Devices),
monitored at 405 nm to measure the time dependent appearance of p-
nitroaniline.
The prothrombinase inhibition assay is performed in a plasma free system
S with modifications to the method described by Sinha, U. et al., Thromb.
Res., 75,
427-436 (1994). Specifically, the activity of the prothrombinase complex is
determined by measuring the time course of thrombin generation using the p-
nitroanilide substrate Chromozym TH. The assay consists of preincubation ( 5
minutes) of selected compounds to be tested as inhibitors with the complex
formed
from factor Xa (0.5 nM), factor Va (2 nM), phosphatidyl serine:phosphatidyl
choline
(25:75, 20 ~.M) in 20 mM Tris~HCl buffer, pH 7.5, containing 0.15 M NaCI, 5 mM
CaCl2 and 0.1% bovine serum albumin. Aliquots from the complex-inhibitor
mixture are added to prothrombin (1 nM) and Chromozym TH (0.1 mM). The rate
of substrate cleavage is monitored at 405 nm for two minutes. Eight different
1 S concentrations of inhibitor are assayed in duplicate. A standard curve of
thrombin
generation by an equivalent amount of untreated complex are used for
determination
of percent inhibition.
Antithrombotic Efficacy in a Rabbit Model of Venous Thrombosis
A rabbit deep vein thrombosis model as described by Hollenbach, S. et al.,
Thromb. Haemost. 71, 357-362 (1994), is used to determine the in-vivo
antithrombotic
activity of the test compounds. Rabbits are anesthetized with LM. inj ections
of Ketamine,
Xylazine, and Acepromazine cocktail. A standardized protocol consists of
insertion of a
thrombogenic cotton thread and copper wire apparatus into the abdominal vena
cava of
the anesthetized rabbit. A non-occlusive thrombus is allowed to develop in the
central
venous circulation and inhibition of thrombus growth is used as a measure of
the
antithrombotic activity of the studied compounds. Test agents or control
saline are
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administered through a marginal ear vein catheter. A femoral vein catheter is
used for
blood sampling prior to and during steady state infusion of test compound.
Initiation of
thrombus formation begins immediately after advancement of the cotton thread
apparatus
into the central venous circulation. Test compounds are administered from time
= 30 min
to time = 1 SO min at which the experiment is terminated. The rabbits are
euthanized and
the thrombus excised by surgical dissection and characterized by weight and
histology.
Blood samples are analyzed for changes in hematological and coagulation
parameters.
Effects of Compounds in Rabbit Venous Thrombosis model
Administration of compounds in the rabbit venous thrombosis model demonstrates
antithrombotic efficacy at the higher doses evaluated. There are no
significant effects of
the compound on the aPTT and PT prolongation with the highest dose (100 ltg/kg
+ 2.57
pg/kg/min). Compounds have no significant effects on hematological parameters
as
compared to saline controls. All measurements are an average of all samples
after steady
state administration of vehicle or (D)-Arg-Gly-Arg-thiazole. Values are
expressed as
mean + SD.
Without further description, it is believed that one of ordinary skill in the
art can,
using the preceding description and the following illustrative examples, make
and utilize
the compounds of the present invention and practice the claimed methods.
