U.S. patent application number 09/796319 was filed with the patent office on 2001-10-18 for benzamide and sulfonamide substitued aminoguanidines and alkoxyguanidines as protese inhibitors.
Invention is credited to Lu, Tianbao, Markotan, Thomas P., Siedem, Colleen, Soll, Richard M., Tomczuk, Bruce E..
Application Number | 20010031870 09/796319 |
Document ID | / |
Family ID | 26763687 |
Filed Date | 2001-10-18 |
United States Patent
Application |
20010031870 |
Kind Code |
A1 |
Soll, Richard M. ; et
al. |
October 18, 2001 |
Benzamide and sulfonamide substitued aminoguanidines and
alkoxyguanidines as protese inhibitors
Abstract
The present invention is directed to aminoguanidine and
alkoxyguanidine compounds, including compounds of Formula I: 1
wherein X is O or NH, L is --O-- or --SO.sub.2--, and
R.sup.1-R.sup.4, R.sup.9-R.sup.19, R.sup.a, R.sup.b, R.sup.c, Y, Z,
n and m are set forth in the specification, as well as hydrates,
solvates or pharmaceutically acceptable salts thereof, that inhibit
proteolytic enzymes such as thrombin. Also described are methods
for preparing the compounds of Formula I. Certain of the compounds
exhibit antithrombotic activity via direct, selective inhibition of
thrombin, or are intermediates useful for forming compounds having
antithrombotic activity. The invention includes a composition for
inhibiting loss of blood platelets, inhibiting formation of blood
platelet aggregates, inhibiting formation of fibrin, inhibiting
thrombus formation, and inhibiting embolus formation in a mammal.
Other uses of compounds of the invention are as anticoagulants
either embedded in or physically linked to materials used in the
manufacture of devices used in blood collection, blood circulation,
and blood storage.
Inventors: |
Soll, Richard M.;
(Lawrenceville, NJ) ; Lu, Tianbao; (Collegeville,
PA) ; Tomczuk, Bruce E.; (Collegeville, PA) ;
Markotan, Thomas P.; (Morgantown, PA) ; Siedem,
Colleen; (Kennett Square, PA) |
Correspondence
Address: |
STERNE, KESSLER, GOLDSTEIN & FOX PLLC
1100 NEW YORK AVENUE, N.W., SUITE 600
WASHINGTON
DC
20005-3934
US
|
Family ID: |
26763687 |
Appl. No.: |
09/796319 |
Filed: |
February 28, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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09796319 |
Feb 28, 2001 |
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09283241 |
Apr 1, 1999 |
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60080568 |
Apr 3, 1998 |
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Current U.S.
Class: |
546/113 ;
544/162; 544/242; 544/59; 546/141; 546/146; 546/153; 546/169 |
Current CPC
Class: |
A61P 9/10 20180101; C07D
217/06 20130101; C07D 213/74 20130101; C07D 211/16 20130101; C07C
279/00 20130101; C07D 213/40 20130101; A61P 29/00 20180101; C07D
295/192 20130101; A61P 43/00 20180101; A61P 1/18 20180101; C07D
211/62 20130101; A61P 7/02 20180101; A61P 9/08 20180101 |
Class at
Publication: |
546/113 ;
546/141; 546/146; 546/153; 546/169; 544/242; 544/162; 544/59 |
International
Class: |
C07D 279/12; C07D
215/20; C07D 217/06 |
Claims
What is claimed is:
1. A compound having the Formula I: 50or a solvate, hydrate or
pharmaceutically acceptable salt thereof; wherein: L represents
--C(O)-- or --SO.sub.2--; R.sup.1 represents a group: 51R.sup.2
represents a group: 52or R.sup.1 and R.sup.2 can be taken together
with the nitrogen atom to which they are attached to form a three
to seven membered ring, either of which contains an additional
nitrogen or oxygen atom, and which is optionally benzo- or
pyrido-fused, said ring being saturated or partially unsatuirated,
and said ring having one or two optional substituents on either a
ring carbon or nitrogen selected from the group consisting of
halogen, hydroxy, acyloxy, alkoxy, aryloxy, alkyl, cycloalkyl,
alkenyl, alkynyl, aryl, aralkyl, heteroaryl,
heteroar(C.sub.1-4)alkyl. carboxyalkyl, alkoxycarbonylalkyl,
hydroxyalkoxyalkyl, cyano(C.sub.2-10)alkyl,
hydroxy(C.sub.2-10)alkyl, alkoxy(C.sub.2-10)alkyl, alkoxyalkyl,
mono alkylamino(C.sub.2-10)alkyl and dialkylamino(C.sub.2-10)alkyl,
carboxy, alkoxycarbonyl, carboxamido, formyl, alkanoyl, aroyl,
aralkanoyl, sulfonyl, alkylsulfonyl, alkoxysulfonyl, and when said
optional substituent is present on a ring carbon.
NR.sup.13N.sup.14; R.sup.12 represents hydrogen, C.sub.3-7
cycloalkyl, C.sub.3-7 cycloalkenyl, C.sub.3-7 heterocycloalkyl,
C.sub.3-7 heterocycloalkenyl, aryl, or heteroaryl, which groups are
optionally substituted with C.sub.1-6 alkyl or hydroxy, or R.sup.12
represents diarylmethyl, diheteroarylmethyl, dicycloalkylmethyl or
(aryl)(heteroaryl)CH--; Z and Z' independently represent a bond, a
C.sub.1-6 alkyl chain, a C.sub.3-6 alkenyl chain, or a C.sub.3-6
alkynyl chain, where one or two nitrogen, oxygen, or sulfur atoms
may be optionally contained within each chain, and the chains are
optionally substituted by one or more groups selected from halogen,
hydroxy, CN, C .sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkoxy(C.sub.1-6)alkyl, C.sub.1-6 acyloxy, NR.sup.13R.sup.14,
NHCOR.sup.15, NHSO.sub.2R.sup.16, COR.sup.15, CO.sub.2R.sup.15,
CONR.sup.13R.sup.14, and SO.sub.2NR.sup.17R.sup.18; provided that
when one of R.sup.1 or R.sup.2 is C.sub.3-8 alkyl, cycloalkyl,
C.sub.3-8 alkenyl, C.sub.3-8alkynyl, aryl, aralkyl, or heteroaryl,
any of which is optionally substituted, then the other of R.sup.1
or R.sup.2 is other than hydrogen, alkyl, aralkyl, aryl,
hydroxy(C.sub.2-10)alkyl, amino(C.sub.2-10)alkyl,
monoalkylamino(C.sub.2-10)alkvl, dialkylamino(C.sub.2-10)alkyl or
carboxyalkyl; R .sup.13-R.sup.16 represent hydrogen, C.sub.1-6
alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.6-10 aryl, mono- or di-hydroxy(C.sub.6-10)aryl, C.sub.6-10
ar(C.sub.1-4)alkyl, pyridyl, pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)-alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono- and
di-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl; or R.sup.13 and R.sup.14
together with the nitrogen to which they are attached form a
C.sub.3-7 heterocycloalkyl ring, or R.sup.16 additionally may
represent trifluoromethyl; R.sup.17 and R.sup.18 are independently
selected from the group consisting of hydrogen, C.sub.1-6 alkyl,
C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.6-10 aryl, C.sub.6-10 ar(C.sub.1-4)alkyl, pyridyl,
pyridyl(C.sub.1-4)alkyl, carboxy(C.sub.1-6)alkyl, C.sub.1-4
alkoxycarbonyl-(C.sub.1-4)alkyl, cyano(C.sub.2-6)alkyl,
hydroxy(C.sub.2-6)alkyl, C.sub.1-4 alkoxy(C.sub.2-6)alkyl, and
mono- and di-(C.sub.1-4)alkylamino(C.sub.2-6)- alkyl, or R.sup.17
and R.sup.18 can be taken together with the nitrogen atom to which
they are attached to form a heterocyclic ring selected from the
group consisting of N-morpholinosulfonyl; N-piperazinylsulfonyl,
optionally substituted in the 4-position with C.sub.1-6 alkyl,
C.sub.1-6 hydroxyalkyl, C.sub.6-10 aryl, C.sub.6-10
aryl(C.sub.1-6)alkyl, C.sub.1-6 alkylsulfonyl, C.sub.6-10
arylsulfonyl, C.sub.1-6 alkylcarbonyl, morpholino or C.sub.6-10
arylcarbonyl; N-pyrrolylsulfonyl; N-piperidinylsulfonyl;
N-pyrrolidinylsulfonyl; N-dihydropyridylsulfonyl;
N-indolylsulfonyl; wherein any of said heterocyclic rings can be
optionally C-substituted; R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are
each independently hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl,
optionally substituted aryl, optionally substituted aralkyl,
optionally substituted heteroaryl, trifluoromethyl, halogen,
hydroxyalkyl, cyano, nitro, carboxamido, --CO.sub.2R.sup.x,
--CH.sub.2OR.sup.x or --OR.sup.x, or when present on adjacent
carbon atoms, R.sup.3 and R.sup.4 may also be taken together to
form --CH.dbd.CH--CH.dbd.CH-- or --(CH.sub.2).sub.q--, where q is
from 2 to 6, and R.sup.5 and R.sup.6 are defined as above; R.sup.x
is hydrogen, alkyl or cycloalkyl wherein said alkyl or cycloalkyl
may optionally have one or more unsaturations; Y is --O--,
--NR.sup.19--, --S--, --CHR.sup.19-- or a covalent bond; R.sup.19,
is hydrogen, C.sub.1-6 alkyl, benzyl, phenyl, C.sub.2-10
hydroxyalkyl, C.sub.2-10 aminoalkyl, C.sub.1-4
monoalkylamino(C.sub.2-8)alkyl, C.sub.1-4
dialkylamino(C.sub.2-8)alkyl or C.sub.2-10 carboxyalkyl; R.sup.7 is
hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl,
monoalkylaminoalkyl, dialkylaminoalkyl, carboxyalkyl, hydroxy,
alkoxy, aralkoxy, aryloxy, heteroaryloxy, or mono- or di-
alkylamino, provided that n is other than zero when R.sup.7 is
hydroxy, alkoxy, aralkoxy, aryloxy, heteroaryloxy, or mono- or di-
alkylamino; R.sup.8, R.sup.9 and R.sup.10 are each independently
hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl,
monoalkylaminoalkyl, dialkylaminoalkyl or carboxyalkyl; or R.sup.7
and R.sup.8 are taken together to form --(CH.sub.2).sub.y--, where
y is zero (a bond), 1 or 2, while R.sup.9 and R.sup.10 are defined
as above; or R.sup.7 and R.sup.10 are taken together to form
--(CH.sub.2).sub.t--, where t is zero (a bond), or 1 to 8, while
R.sup.8 and R.sup.9 are defined as above; or R.sup.8 and R.sup.9
are taken together to form --(CH.sub.2).sub.r--, where r is 2-8,
while R.sup.7 and R.sup.10 are defined as above; X is oxygen or NH;
R.sup.11 is hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl,
aminoalkyl, monoalkylamino(C.sub.2-10)alkyl,
dialkylamino(C.sub.2-10)alkyl or carboxyalkyl; R.sup.a, R.sup.b and
R.sup.c are independently hydrogen, alkyl, hydroxy, alkoxy,
aryloxy, aralkoxy, alkoxycarbonyloxy, cyano or --CO.sub.2R.sup.w;
R.sup.w is alkyl, trichloroethyl, cycloalkyl, phenyl, benzyl,
53where R.sup.d and R.sup.e are independently hydrogen, C.sub.1-6
alkyl, C.sub.2-6 alkenyl or phenyl, R.sup.f is hydrogen, C.sub.1-6
alkyl, C.sub.2-6 alkenyl or phenyl, R.sup.g is hydrogen, C.sub.1-6
alkyl, C.sub.2-6 alkenyl or phenyl, and R.sup.h is aralkyl or
C.sub.1-6 alkyl; n is from zero to 8; and m is from zero to 4.
2. A compound ofclaim 1, wherein the moiety --L--NR.sup.1R.sup.2 is
a ttached to the benzene ring in the meta-position.
3. A compound having the structure of Formula Ia: 54wherein:
R.sup.1 represents a group: 55R.sup.2 represents a group: 56or
R.sup.1 and R.sup.2 can be taken together with the nitrogen atom to
which they are attached to form a three to seven membered ring,
either of which contains an additional nitrogen or oxygen atom, and
which is optionally benzo- or pyrido-fused, said ring being
saturated or partially unsaturated, and said ring having one or two
optional substituents on either a ring carbon or nitrogen selected
from the group consisting ofhalogen, hydroxy, acyloxy, alkoxy,
aryloxy, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, heteroar(C.sub.1-4)alkyl, carboxyalkyl,
alkoxycarbonylalkyl, hydroxyalkoxyalkyl, cyano(C.sub.2-10)alkyl,
hydroxy(C.sub.2-10)alkyl, alkoxy(C.sub.2-10)alkyl, alkoxyalkyl,
monoalkylamino(C.sub.2-10)alkyl and dialkylamio(C.sub.2-10) alkyl,
carboxy, alkoxycarbonyl, carboxamido, formyl, alkanoyl, aroyl,
aralkanoyl, sulfonyl, alkylsulfonyl, alkoxysulfonyl, and when said
optional substituent is present on a ring carbon,
NR.sup.13R.sup.14; R.sup.12 represents hydrogen, C.sub.3-7
cycloalkyl, C.sub.3-7 cycloalkenyl, C.sub.3-7 heterocycloalkyl,
C.sub.3-7 heterocycloalkenyl, aryl, or heteroaryl, which groups are
optionally substituted with C.sub.1-6 alkyl or hydroxy, or R.sup.12
represents diarylmethyl, diheteroarylmethyl, dicycloalkylmethyl or
(aryl)(heteroaryl)CH--; Z and Z' independently represent a bond, a
C.sub.1-6 alkyl chain, a C.sub.3-6 alkenyl chain, or a C.sub.3-6
alkynyl chain, where one or two nitrogen, oxygen, or sulfur atoms
may be optionally contained within each chain, and the chains are
optionally substituted by one or more groups selected from halogen,
hydroxy, CN, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkoxy(C.sub.1-6)alkyl, C.sub.1-6 acyloxy, NR.sup.13R.sup.14,
NHCOR.sup.15, NHSO.sub.2R.sup.16, COR.sup.15, CO.sub.2R.sup.15,
CONR.sup.13R.sup.14, and SO.sub.2NR.sup.17R.sup.18: provided that
when one of R.sup.1 or R.sup.2 is C.sub.3-8 alkyl, cycloalkyl,
C.sub.3-8 alkenyl, C.sub.3-8 alkynyl, aryl, aralkyl, or heteroaryl,
any of which is optionally substituted, then the other of R.sup.1
or R.sup.2 is other than hydrogen, alkyl, aralkyl. aryl,
hydroxy(C.sub.2-10)alkyl, amino(C.sub.2-10,)alkyl,
monoalkylamino(C.sub.2-10)alkyl, dialkylamino(C.sub.2-10)alkyl or
carboxyalkyl; R.sup.13-R.sup.16 represent hydrogen, C.sub.1-6
alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.6-10 aryl, mono- or di-hydroxy(C.sub.6-10)aryl, C.sub.6-10
ar(C.sub.1-4)alkyl, pyridyl, pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)-alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono (C.sub.1-4)alkylamino(C.sub.2-6)alkyl
and di(C.sub.1-4)alkylamino(C.sub.2- -6)alkyl; or R.sup.13 and
R.sup.14 together with the nitrogen to which they are attached form
a C.sub.3-7 heterocycloalkyl ring; or R.sup.16 additionally may
represent trifluoromethyl; R.sup.17 and R.sup.18 are independently
selected from the group consisting of hydrogen, C.sub.1-6 alkyl,
C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.6-10 aryl, C.sub.6-10 ar(C.sub.1-4)alkyl, pyridyl,
pyridyl(C.sub.1-4)alkyl, carboxy(C.sub.1-6)alkyl, C.sub.1-4
alkoxycarbonyl-(C.sub.1-4)alkyl, cyano(C.sub.2-6)alkyl,
hydroxy(C.sub.2-6)alkyl, C.sub.1-4 alkoxy(C.sub.2-6)alkyl, and
mono(C.sub.1-4)alkylamino(C.sub.2-6)alkyl and
di(C.sub.1-4)alkylamino(C.s- ub.2-6)alkyl, or R.sup.17 and R.sup.18
can be taken together with the nitrogen atom to which they are
attached to form a heterocyclic ring selected from the group
consisting of N-morpholinosulfonyl; N-piperazinylsulfonyl
optionally substituted in the 4-position with C.sub.1-6 alkyl,
C.sub.1-6 hydroxyalkyl, C.sub.6-10 aryl, C.sub.6-10
aryl(C.sub.1-6)alkyl, C.sub.1-6 alkylsulfonyl, C.sub.6-10
arylsulfonyl, C.sub.1-6 alkylcarbonyl, morpholino or C.sub.6-10
arylcarbonyl; N-pyrrolylsulfonyl; N-piperidinylsulfonyl;
N-pyrrolidinylsulfonyl; N-dihydropyridylsulfonyl;
N-indolylsulfonyl; wherein any of said heterocyclic rings can be
optionally C-substituted; R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are
each independently one of hydrogen. alkyl, cycloalkyl, alkenyl,
alkynyl, optionally substituted aryl, optionally substituted
aralkyl, optionally substituted heteroaryl, trifluoromethyl,
halogen, hydroxyalkyl, cyano, nitro, carboxamido,
--CO.sub.2R.sup.x, --CH.sub.2OR.sup.x or --OR.sup.x, or when
present on adjacent carbon atoms, R.sup.3 and R.sup.4 may also be
taken together to form one of --CH.dbd.CH--CH.dbd.CH-- or
--(CH.sub.2).sub.q--, where q is from 2 to 6, and R.sup.5 and
R.sup.6 are defined as above; R.sup.x is hydrogen, alkyl or
cycloalkyl wherein said alkyl or cycloalkyl groups may optionally
have one or more unsaturations; Y is --O--, --NR.sup.19--, --S--,
--CHR.sup.19-- or a covalent bond; R.sup.19 is hydrogen, C.sub.1-6
alkyl, benzyl, phenyl, C.sub.2-10 hydroxyalkyl, C.sub.2-10
aminoalkyl, C.sub.1-4 monoalkylamino(C.sub.2-8)alkyl, C.sub.1-4
dialkylamino(C.sub.2-8)alkyl or C.sub.2-10 carboxyalkyl; R.sup.7 is
hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl,
monoalkylaminoalkyl, dialkylaminoalkyl, carboxyalkyl, hydroxy,
alkoxy, aralkoxy, aryloxy, heteroaryloxy, or mono- or di-
alkylamino, provided that n is other than zero when R.sup.7 is
hydroxy, alkoxy, aralkoxy, aryloxy, heteroaryloxy, or mono- or di-
alkylamino; R.sup.8, R.sup.9 and R.sup.10 are independently
hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl, aminoalkyl,
monoalkylaminoalkyl, dialkylaminoalkyl or carboxyalkyl; or R.sup.7
and R.sup.8 are taken together to form --CH.sub.2).sub.y--, where y
is zero (a bond), 1 or 2, while R.sup.9 and R.sup.10 are defined as
above; or R.sup.7 and R.sup.10 are taken together to form
--(CH.sub.2).sub.t--, where t is zero (a bond), or 1 to 8, while
R.sup.8 and R.sup.9 are defined as above; or R.sup.8 and R.sup.9
are taken together to form --(CH.sub.2).sub.r--, where r is 2-8,
while R.sup.7 and R.sup.10 are defined as above; X is oxygen or NH;
R.sup.11 is hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl,
aminoalkyl, monoalkylarnino(C.sub.2-10)alkyl,
dialkylamino(C.sub.2-10)alk- yl or carboxyalkyl; R.sup.a, R.sup.b
and R.sup.c are independently hydrogen, alkyl, hydroxy, alkoxy,
aryloxy, aralkoxy, alkoxycarbonyloxy, cyano or --CO.sub.2R.sup.w;
R.sup.w is alkyl, trichloroethyl, cycloalkyl, phenyl, benzyl,
57where R.sup.d and R.sup.e are independently hydrogen, C.sub.1-6
alkyl, C.sub.2-.sub.6 alkenyl or phenyl, R.sup.f is hydrogen,
C.sub.1-6 alkyl, C.sub.2-6 alkenyl or phenyl, R.sup.g is hydrogen,
C.sub.1-6 alkyl, C.sub.2-6 alkenyl or phenyl, and R.sup.h is
aralkyl or C.sub.1-6 alkyl; n is from zero to 8; and m is from zero
to 4.
4. A compound of claim 3, wherein R.sup.2 represents a group
58where Z' is C.sub.3-6 alkenyl or C.sub.1-6 alkyl; and R.sup.12 is
hydrogen, C.sub.3-7 heterocycloalkyl, aryl, or heteroaryl.
5. A compound of claim 4, wherein Z' is allyl, methyl, ethyl,
propyl, or pentyl.
6. A compound of claim 4, wherein Z' contains an oxygen group
within the chain.
7. A compound of claim 4, wherein Z' contains an oxygen group
within the chain, and said chain is substituted by a group selected
from hydroxy, C.sub.1-6 alkoxy, NHSO.sub.2R.sup.16,
CO.sub.2R.sup.15, CONR.sup.13R.sup.14, or
SO.sub.2NR.sup.17R.sup.18.
8. A compound of claim 4, wherein R.sup.12 is pyrrolidine,
morpholine, phenyl substituted by CO.sub.2R.sup.15, oxadiazole
substituted by hydroxy, triazole, or tetrazole substituted by
C.sub.1-6 alkyl.
9. A compound of claim 3, wherein R.sup.1 represents a group 59Z is
a bond or C.sub.1-6 alkyl group, and R.sup.12 is hydrogen,
C.sub.3-7 cycloalkyl, aryl, or heteroaryl.
10. A compound of claim 3, wherein Z represents a bond, and
R.sup.12 is C.sub.3-7 cycloalkyl or phenyl substituted with
C.sub.1-6 alkyl or hydroxy.
11. A compound of claim 10, wherein R.sup.12 is cyclobutyl,
cyclopentyl, cyclohexyl, diphenylmethyl or dicyclohexylmethyl.
12. A compound of claim 11, wherein Z represents a C.sub.1-4 alkyl
group, and R.sup.12 is hydrogen, cycloalkyl or heteroaryl.
13. A compound of claim 3, wherein R.sup.3 is hydrogen, C.sub.1-3
alkyl, to halogen, or C.sub.1-2 alkoxy.
14. A compound of claim 3, wherein R.sup.4, R.sup.5 and R.sup.6
independently represent hydrogen or halogen.
15. A compound of claim 3, wherein Y is divalent oxygen (--O--),
--NR.sup.19-- or a covalent bond.
16. A compound of claim 3, wherein R.sup.19 is hydrogen, C.sub.1-6
alkyl or C.sub.3-6 cycloalkyl.
17. A compound of claim 3, wherein R.sup.11 is hydrogen or
C.sub.1-6 alkyl.
18. A compound of claim 3, wherein R.sup.7, R.sup.8, R.sup.9 and
R.sup.10 are independently one of hydrogen, C.sub.1-6 alkyl,
C.sub.6-10 ar(C.sub.1-6)alkyl, C.sub.6-10 aryl, C.sub.2-10
hydroxyalkyl or C.sub.2-7 carboxyalkyl.
19. A compound of claim 18, wherein R.sup.7, R.sup.8, R.sup.9 and
R.sup.10 are independently hydrogen, methyl, ethyl, propyl,
n-butyl, benzyl, phenylethyl, 2-hydroxyethyl, 3-hydroxypropyl,
4-hydroxybutyl, 2-carboxymethyl, 3-carboxyethyl and
4-carboxypropyl.
20. A compound of claim 3, wherein R.sup.7 and R.sup.8 are taken
together to form --(CH.sub.2).sub.y-- and y is 2.
21. A compound of claim 3, wherein R.sup.8 and R.sup.9 are taken
together to form --(CH.sub.2).sub.r-- and r is 2.
22. A compound of claim 3, wherein X is O.
23. A compound of claim 3, wherein R.sup.a, R.sup.b and R.sub.c are
independently hydrogen, hydroxy, C.sub.1-6 alkyl, C.sub.1-6 alkoxy,
cyano or --CO.sub.2R.sup.w, where R.sup.w, in each instance, is
selected from the group consisting of C.sub.1-4 alkyl,
trichloroethyl, C.sub.4-7 cycloalkyl and benzyloxycarbonyl.
24. A compound of claim 23, wherein R.sup.a, R.sup.b and R.sup.c
are independently selected from the group consisting of hydrogen,
methyl, ethyl, propyl, n-butyl, hydroxy, methoxy, ethoxy, cyano,
--CO.sub.2CH.sub.3, --CO.sub.2CH.sub.2CH.sub.3 and
--CO.sub.2CH.sub.2CH.sub.2CH.sub.3.
25. A compound of claim 24, wherein R.sup.a, R.sup.b and R.sup.c
are each hydrogen.
26. A compound of claim 3, wherein R.sup.a, R.sup.b and R.sup.c are
independently --CO.sub.2R.sup.w, where R.sup.w is one of 60where
R.sup.d, R.sup.e, R.sup.f, R.sup.g and R.sup.h are as defined in
claim 3.
27. A compound of claim 3, wherein R.sup.d, R.sup.e and R.sup.g are
each hydrogen, R.sup.f is methyl, and R.sup.h is benzyl or
tert-butyl.
28. A compound of claim 3, wherein n is zero, 1 or 2.
29. A compound of claim 3, wherein m is zero, 1, 2 or 3.
30. A compound of claim 3, wherein R.sup.1 and R.sup.2 are taken
together with the nitrogen to which they are attached to form a
C.sub.3-7 heterocycloalkyl or C.sub.3-7 heterocycloalkenyl group,
optionally benzo fused and optionally including an oxygen atom or
an additional nitrogen atom, and which may be optionally
substituted by C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy,
C.sub.2-6 alkoxycarbonyl, formyl, (C.sub.6-10)ar(C.sub.1-4)alkyl,
C.sub.6-10 aryl, pyridyl, hydroxyalkoxyalkyl, halogen, or
NR.sup.13R.sup.14; R.sup.13 and R.sup.14 independently represent
hydrogen, C.sub.1-6 alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.6-10 aryl, C.sub.6-10 ar(C.sub.1-4)alkyl,
pyridyl, pyridyl(C.sub.1-4)alkyl, carboxy(C.sub.1-6)-alkyl,
C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl, cyano(C.sub.2-6)alkyl,
hydroxy(C.sub.2-6)alkyl, C.sub.1-4 alkoxy(C.sub.2-6)alkyl, mono-
and di-(C.sub.1-4)alkylamino(C.sub.2-6)alky- l; or R.sup.13 and
R.sup.14 together with the nitrogen to which they are attached form
a C.sub.3-7 heterocycloalkyl ring; R.sup.3 is hydrogen, C.sub.1-3
alkyl, halogen or C.sub.1-2 alkoxy; R.sup.4, R.sup.5 and R.sup.6
are hydrogen or halogen; Y is --O--; R.sup.a, R.sup.b and R.sup.c
are each one of hydrogen, C.sub.1-4 alkyl, hydroxy, C.sub.1-4
alkoxy, phenoxy, C.sub.1-4 alkyloxycarbonyl, benzyloxycarbonyl,
cyano, 61where R.sup.h is benzyl, methyl, ethyl, isopropyl,
sec-butyl or t-butyl, and where R.sup.f is hydrogen or C.sub.1-6
alkyl; R.sup.11 is one of hydrogen, C.sub.1-6 alkyl, C.sub.6-10
ar(C.sub.1-6)alkyl, C.sub.6-10 aryl, C.sub.2-10 hydroxyalkyl,
C.sub.2-10 aminoalkyl, mono(C.sub.1-4)alkylamino(C.sub.2-8)alkyl,
di(C.sub.1-4)alkylamino(C.sub.- 2-8)alkyl or C.sub.2-10
carboxyalkyl; R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are
independently one of hydrogen, C.sub.1-6 alkyl, C.sub.2-10
carboxyalkyl or C.sub.2-10 hydroxyalkyl, or R.sup.7 and R.sup.8 are
taken together to form --(CH.sub.2).sub.y-- where y is zero, 1 or
2, while R.sup.9 and R.sup.10 are defined as above; or R.sup.7 and
R.sup.10 are taken together to form --(CH.sub.2).sub.t--, where t
is zero (a bond), or 1, 2 or 3, while R.sup.8 and R.sup.9 are
defined as above; or R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r--, where r is 2, 3, or 4, while R.sup.7 and
R.sup.10 are defined as above; n is from zero to 4; and m is from
zero to 4.
31. A compound of claim 3, wherein R.sup.1 is C.sub.3-7 cycloalkyl
or C.sub.3-7 cycloalkenyl, either of which is optionally
substituted by C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy, halogen,
carboxylic acid, a C.sub.1-4 carboxylic acid ester group, or
NR.sup.13R.sup.14, and R.sup.2 is C.sub.3-6 alkenyl, or C.sub.3-6
alkynyl, either of which is optionally substituted by C.sub.1-6
alkyl, hydroxy. C.sub.1-4 alkoxy, halogen, carboxylic acid, a
C.sub.1-4 carboxylic acid ester group, or NR.sup.13R.sup.14;
R.sup.13 and R.sup.14 independently represent hydrogen, C.sub.1-6
alkyl, C.sub.3-7 cycloalkyl. C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.6-10 aryl, C.sub.6-10 ar(C.sub.1-4)alkyl, pyridyl,
pyridyl(C.sub.1-4)alkyl. carboxy(C.sub.1-6)-alkyl, C.sub.1-4
alkoxycarbonyl(C.sub.1-4)alkyl, cyano(C.sub.2-6)alkyl;
hydroxy(C.sub.2-6)alkyl, C.sub.1-4 alkoxy(C.sub.2-6)alkyl, mono-
and di-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl; or R.sup.13 and
R.sup.14 together with the nitrogen to which they are attached form
a C.sub.3-7 heterocycloalkyl ring; R.sup.3 is hydrogen, C.sub.1-3
alyl, halogen or C.sub.1-2 alkoxy; R.sup.4, R.sup.5 and R.sup.6 are
hydrogen or halogen; Y is --O--; R.sup.a, R.sup.b and R.sup.c are
each one of hydrogen, C.sub.1-4 alkyl, hydroxy, Cl.sub.1 alkoxy,
phenoxy, C.sub.1-4 alkyloxycarbonyl, benzyloxycarbonyl, cyano,
62where R.sup.h is benzyl, methyl, ethyl, isopropyl, sec-butyl or
t-butyl, and where R.sup.f is hydrogen or C.sub.1-6 alkyl; R.sup.11
is one of hydrogen, C.sub.1-6 alkyl, C.sub.6-10 ar(C.sub.1-6)alkyl,
C.sub.6-10 aryl. C.sub.2-10 hydroxyalkyl, C.sub.2-10 aminoalkyl,
mono(C.sub.1-4)alkylamino(C.sub.2-8)- alkyl.
di(C.sub.1-4)alkylamino(C.sub.2-8)alkyl or C.sub.2-10 carboxyalkyl;
R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are independently one of
hydrogen, C.sub.1-6 alkyl, C.sub.2-10 carboxyalkyl or C.sub.2-10
hydroxyalkyl, or R.sup.7 and R.sup.8 are taken together to form
--(CH.sub.2).sub.y-- where y is zero, 1 or 2, while R.sup.9 and
R.sup.10 are defined as above; or R.sup.7 and R.sup.10 are taken
together to form --(CH.sub.2).sub.t--, where t is zero (a bond), or
1, 2 or 3, while R.sup.8 and R.sup.9 are defined as above; or
R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r--, where r is 2, 3, or 4, while R.sup.7 and
R.sup.10 are defined as above; n is from zero to 4; and m is from
zero to 4.
32. A compound of claim 3, wherein R.sup.1 is C.sub.3-7
heterocycloalkyl(C.sub.1-6)alkyl, C.sub.3-7
heterocycloalkenyl(C.sub.1-6)- alkyl. heteroaryl(C.sub.1-6)alkyl,
C.sub.3-7 heterocycloalkyl(C.sub.3-6)al- kenyl, C.sub.3-7
heterocycloalkenyl(C.sub.3-6)alkenyl, heteroaryl(C.sub.3-6)alkenyl,
C.sub.3-7 heterocycloalkyl(C.sub.3-6)alkyny- l, C.sub.3-7
heterocycloalkenyl(C.sub.3-6)alkynyl, heteroaryl(C.sub.3-6)al-
kynyl, di(C.sub.5-10 aryl)(C.sub.1-3)alkyl, di(C.sub.3-8
cycloalkyl)(C.sub.1-3)alkyl or di(C.sub.3-8
cycloalkenyl)(C.sub.1-3)alkyl- , any of which is optionally
substituted by C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy, halogen,
carboxylic acid, a C.sub.1-4 carboxylic acid ester group, or
NR.sup.13R.sup.14; R.sup.13 and R.sup.14 independently represent
hydrogen, C.sub.1-6 alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.6-10 aryl, C.sub.6-10 ar(C.sub.1-4)alkyl,
pyridyl, pyridyl(C.sub.1-4)alkyl, carboxy(C.sub.1-6)-alkyl,
C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl, cyano(C.sub.2-6)alkyl,
hydroxy(C.sub.2-6)alkyl, C.sub.1-4 alkoxy(C.sub.2-6)alkyl, mono-
and di- (C.sub.1-4)alkylamino(C.sub.2-6)alk- yl; or R.sup.13 and
R.sup.14 together with the nitrogen to which they are attached form
a C.sub.3-7 heterocycloalkyl ring; R.sup.3 is hydrogen, C.sub.1-3
alkyl, halogen or C.sub.1-2 alkoxy; R.sup.4, R.sup.5 and R.sup.6
are hydrogen or halogen; Y is --O--; R.sup.a, R.sup.b and R.sup.c
are each one of hydrogen, C.sub.1-4 alkyl, hydroxy, C.sub.1-4
alkoxy, phenoxy, C.sub.1-4 alkyloxycarbonyl, benzyloxycarbonyl,
cyano, 63where R.sup.h is benzyl, methyl, ethyl, isopropyl,
sec-butyl or t-butyl, and where R.sup.f is hydrogen or C.sub.1-6
alkyl; R.sup.11 is one of hydrogen, C.sub.1-6 alkyl, C.sub.6-10
ar(C.sub.1-6)alkyl, C.sub.6-10 aryl, C.sub.2-10 hydroxyalkyl,
C.sub.2-10 aminoalkyl, mono(C.sub.1-4)alkylamino(C.sub.2-8)alkyl,
di(C.sub.1-4)alkylamino(C.sub.- 2-8)alkyl or C.sub.2-10
carboxyalkyl; R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are
independently one of hydrogen, C.sub.1-6 alkyl, C.sub.2-10
carboxyalkyl or C.sub.2-10 hydroxyalkyl, or R.sup.7 and R.sup.8 are
taken together to form --(CH.sub.2).sub.r-- where y is zero, 1 or
2, while R.sup.9 and R.sup.10 are defined as above; or R.sup.7 and
R.sup.10 are taken together to form --(CH.sub.2).sub.t--, where t
is zero (a bond), or 1, 2 or 3, while R.sup.8 and R.sup.9 are
defined as above; or R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r--, where r is 2, 3, or 4, while R.sup.7 and
R.sup.10 are defined as above; n is from zero to 4; and m is from
zero to 4.
33. A compound of claim 3 having the Formnula IIa: 64or a solvate,
hydrate, pharmaceutically acceptable salt or prodrug thereof,
wherein: R.sup.1A represents a group: 65wherein Z.sup.A represents
a bond or C.sub.1-6 alkyl; and R.sup.12A represents hydrogen,
C.sub.3-7 cycloalkyl, C.sub.1-6 alkoxy, aryl optionally substituted
by halogen, hydroxy. heteroaryl, diphenylmethyl or
dicyclohexylmethyl; R.sup.2A represents a group: 66wherein Z.sup.B
represents C.sub.3-6 alkenyl or C.sub.1-6 alkyl optionally
substituted by CO.sub.2R.sup.5 or COR.sup.15; R.sup.12B represents
hydrogen, C.sub.1-6 alkoxy, mono- or di- C.sub.1-3 alkylamino,
phenyl substituted by CO.sub.2R.sup.15, oxadiazole substituted by a
hydroxy group, or an unsubstituted C-linked tetrazole group; and
R.sup.15 is C.sub.1-3 alkyl or mono- or di-hydroxyphenyl; or
R.sup.1A and R.sup.2A can be taken together with the nitrogen atom
to which they are attached to form a three to seven membered ring,
either of which contains an additional nitrogen or oxygen atom, and
which is optionally benzo- or pyrido-fused, said ring being
saturated or partially unsaturated, and said ring having one or two
optional substituents on either a ring carbon or nitrogen selected
from the group consisting of halogen, hydroxy, acyloxy, alkoxy,
aryloxy, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, heteroar(C.sub.1-4)alkyl, carboxyalkyl,
alkoxycarbonylalkyl, hydroxyalkoxyalkyl, cyano(C.sub.2-10)alkyl,
hydroxy(C.sub.2-10)alkyl, alkoxy(C.sub.2-10)alkyl, alkoxyalkyl,
monoalkylarnino(C.sub.2-10)alkyl and dialkylamino(C.sub.2-10)alkyl,
carboxy, alkoxycarbonyl, carboxamido, formyl, alkanoyl, aroyl,
aralkanoyl, sulfonyl, alkylsulfonyl, alkoxysulfonyl, and when said
optional substituent is present on a ring carbon,
NR.sup.13N.sup.14; R.sup.13 and R.sup.14 independently represent
hydrogen, C.sub.1-6 alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.6-10 aryl, hydroxy(C.sub.6-10)aryl,
C.sub.6-10 ar(C.sub.1-4)alkyl, pyridyl, pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)-alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl,
or di- (C.sub.1-4)alkylamino(C.sub.2-6)alkyl, or R.sup.13 and
R.sup.14 together with the nitrogen to which they are attached form
a C.sub.3-7 heterocycloalkyl ring; R.sup.3A represents C.sub.1-3
alkyl or halogen; R.sup.11A represents hydrogen, C.sub.6-10
ar(C.sub.1-4) alkyl or C.sub.1-4 alkyl; X.sup.A is oxygen or NH;
R.sup.a, R.sup.b and R.sup.c are hydrogen; and a is from zero to
8.
34. A compound of claim 33, wherein X.sup.A is oxygen.
35. A compound of claim 33, wherein: R.sup.1A represents a group:
67wherein Z.sup.A represents a bond or C.sub.1-6 alkyl; and
R.sup.12A represents hydrogen, C.sub.3-7 cycloalkyl, C.sub.1-6
alkoxy, aryl optionally substituted by halogen or hydroxy, or
heteroaryl; R.sup.2A represents a group: 68wherein Z.sup.B
represents C.sub.3-6 alkenyl or C.sub.1-6 alkyl optionally
substituted by CO.sub.2R.sup.15 or COR.sup.15; R.sup.12B represents
hydrogen, C.sub.1-6 alkoxy, or mono- or di- C.sub.1-3 alkylamino;
and R.sup.15 is C.sub.1-3 alkyl or mono- or di-hydroxyphenyl; or
R.sup.1A and R.sup.2A are taken together with the nitrogen to which
they are attached to form a C.sub.3-7 heterocycloalkyl or C.sub.3-7
heterocycloalkenyl group, optionally benzo fused and optionally
including an oxygen atom or an additional nitrogen atom, and which
may be optionally substituted by C.sub.1-6 alkyl, hydroxy,
C.sub.1-4 alkoxy, C.sub.2-6 alkoxycarbonyl, formyl,
(C.sub.6-10)ar(C.sub.1-4)alkyl, C.sub.6-10 aryl, pyridyl,
hydroxy(C.sub.1-4)alkoxy(C.sub.1-4)alkyl, halogen, or
NR.sup.13R.sup.14; R.sup.13 and R.sup.14 independently represent
hydrogen, C.sub.1-6 alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl,
C.sub.2-6 alkynyl, C.sub.6-10 aryl, hydroxy(C.sub.6-10)aryl,
C.sub.6-10 ar(C.sub.1-4)alkyl, pyridyl, pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)-alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl,
or di- (C.sub.1-4)alkylamino(C.sub.2-6)alkyl; or R.sup.13 and
R.sup.14 together with the nitrogen to which they are attached form
a C.sub.3-7 heterocycloalkyl ring; R.sup.3A represents halogen;
X.sup.A is --O--; R.sup.11A is hydrogen, C.sub.6-10 ar(C.sub.1-4)
alkyl or C.sub.1-4 alkyl; R.sup.a, R.sup.b and R.sup.c are
hydrogen; and a is 1.
36. A compound of claim 35, wherein R.sup.3A is methyl.
37. A compound of claim 35, wherein R.sup.3A is chloro.
38. Acompoundofclaim33,whichis
[3-{5-chloro-3-(N-cyclopentyl-N-[prop-2-eny-
l]aminocarbonyl)phenoxy}propoxyamino]carboxamidine
hydrochloride.
39. A compound of claim 33, which is one of
[3-{5-chloro-3-(N-cyclopentyl-- N-[prop-2-eny ]aminocarbonyl)
phenoxy}propoxyamino]carboxamidine hydrochloride;
[3-{5-chloro-3-(4-benzylpiperidinylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate; [3-{5
-chloro-3 -(N,N-bis[2-methoxyethyl]aminocarbonyl)
phenoxy}propoxyamino]carboxamidin- e trifluoroacetate;
[3-{5-chloro-3 -(N-methyl-N-[2-{2-pyridyl }
ethyl]aminocarbonyl)phenoxy }propoxyamino]carboxamidine
trifluoroacetate; [3-{5-chloro-3-(N-methyl-N-[3-pyridylmethyl
]aminocarbonyl) phenoxy}propoxyamino]carboxamidine
trifluoroacetate;
[3-{5-chloro-.sup.3-(N-ethyl-N-[4-pyridylmethyl]aminocarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate; ethyl
2-[5-{3-(amidinoaminooxy)propoxy}-3-chlorophenyl]-N-{2-pyridylmethyl}amin-
ocarbonyl]acetate trifluoroacetate; methyl
2-[5-{3-(amidinoaminooxy)propox-
y}-3-chlorophenyl]-N-{2-pyridylmethyl}aminocarbonyl]acetate
trifluoroacetate; [3-{5-chloro-3-([2- {3,4-dihydroxyphenyl
}-2-oxoethyl]-N-methyl
aminocarbonyl)phenoxy}propoxyamino]carboxamidine trifluoroacetate;
[3-{5-chloro-3-(N-[2-{dimethylamino}ethyl]-N-ethylamnin-
ocarbonyl)phenoxy}propoxyamino]carboxamidine trifluoroacetate;
[3-{5-chloro-3-(4-formylpiperazinylcarbonyl)
phenoxy}propoxyamino]carboxa- midine trifluoroacetate;
[3-{5-chloro-3-(4-phenylpiperazinycarbonyl)
phenoxylpropoxyamino]carboxamidine trifluoroacetate;
[3-5-chloro-3-4-benzypperaznycarbony
phenoxylpropoxyamnino]carboxamidine trifluoroacetate;
[3-5-chloro-3-(N,N-dimethylaminocarbonyl
phenoxy}propoxyamino]carboxarnidine trifluoroacetate; [3
-{5-chloro-3)-(piperidinylcarbonyl)phenoxy}
propoxyamino]carboxamidine trifluoro acetate;
[3-{5-chloro-3-(4-[2-pyridyl]piperaznylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoro acetate;
[3-{5-chloro-3-(4-[4-pyridyl]piperazinylcarbonyl)
phenoxy}propoxyamino]ca- rboxarnidine trifluoroacetate;
[3-{5-chloro -3-(2-[1,2,3,4-tetrahydro]isoq- uinolinylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate;
[3-{5-chloro-3-(azaperhydroepinylcarbonyl)
phenoxyl}propoxyamino]carboxam- idine trifluoroacetate; ethyl
3-({5-[3-(amidinoaminooxy)propoxy]-3-chlorop-
henyl}-N-benzylcarbonylamino)propanoate trifluoroacetate; ethyl 1-(
{5-[3-(amidinoaminooxy)propoxy]-3 -chlorophenyl
}carbonyl)piperidine-4-ca- rboxylate trifluoroacetate;
[3-{5-chloro-3-(morpholin-4-ylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate; and methyl 2-(
{5-
[3-(amidinoaminooxy)propoxy]-3-chlorophenyl}-N-methylcarbonylamino)acetat-
e trifluoroacetate.
40. The process for preparing a hydroxyguanidine compound of claim
1, comprising reacting an alkoxyamine compound of the Formula IX:
69wherein R.sup.1-R.sup.6, Y, n, m, R.sup.7, R.sup.8, R.sup.9 and
R.sup.10 are defined as in claim 1, with a guanidinylating
reagent.
41. The process of claim 40, wherein the guanidinylating reagent is
selected from the group consisting of aminoiminosulfonic acid,
substituted 1 H-pyrazole-1-carboxamidines, and
N,N'-bis(tert-butoxycarbon- yl) S-methyl isothiourea.
42. A process for preparing an aminoguanidine compound of claim 1,
comprising reacting an aminoguanidine of the Formula VII: 70wherein
R.sup.11, R.sup.a, R.sup.b and R.sup.c are as defined in claim 1,
with a carbonyl-containing compound of the Formula VIII: 71wherein
R.sup.1-R.sup.6, Y, n, m, R.sup.7, R.sup.8, R.sup.9 and R.sup.10
are as defined in claim 1, to form an amidinohydrazone; and
thereafter selectively reducing the hydrazone carbon to nitrogen
double bond of the amidinohydrazone.
43. The process of claim 42, wherein the aminoguanidine is provided
as a salt.
44. The process of claim 42, wherein the aminoguanidine is provided
as a nitrate salt.
45. The process of claim 42, wherein (a) the first step proceeds at
ambient temperature using alcohol as a solvent; and (b) 4 N HCl in
dioxane is added to the reaction mixture.
46. A pharmaceutical composition for inhibiting proteolysis in a
mammal, comprising an amount of a compound of claim 1 effective to
inhibit proteolysis, and a pharmaceutically acceptable carrier or
diluent.
47. The pharmaceutical composition of claim 46, comprising an
amount of said compound effective to inhibit a trypsin-like
protease.
48. A method of inhibiting proteolysis in a mammal, comprising
administering to the mammal a composition of claim 46.
49. The method of claim 48, wherein a trypsin-like protease is
inhibited.
50. A method of treating pancreatitis, thrombosis, ischemia,
stroke, restenosis, emphysema or inflammation in a mammal,
comprising administering to the mammal a composition of claim
46.
51. A method of inhibiting thrombin-induced platelet aggregation
and clotting of fibrinogen in plasma, comprising administering to
the mammal a composition of claim 46.
52. A method for inhibiting thrombin in blood comprising adding to
the blood a compound of claim 1.
53. A method for inhibiting formation of blood platelet aggregates
in blood comprising adding to the blood a compound of claim 1.
54. A method for inhibiting thrombus formation in blood comprising
adding to the blood a compound of claim 1.
55. In a device used in blood collection, blood circulation, and
blood storage wherein said device includes an effective amount of a
thrombin inhibiting compound or macromolecule as an anticoagulant,
either embedded in, or physically linked to, one or more materials
that form the structure of said device, the improvement comprising
employing as said thrombin inhibitor one or more compounds as
claimed in claim 1.
56. The device of claim 55, wherein said device is a catheter,
blood dialysis machine, blood collection syringe, blood collection
tube, blood line or extracorporeal blood circuit.
57. The device of claim 55, wherein said device is a stent that can
be surgically inserted into a mammal.
Description
[0001] This application claims the benefit, under 35 U.S.C. .sctn.
119(e), of the earlier filing date of U.S. Provisional Application
No. 60/080,568, filed on Apr. 3, 1998.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates to novel compounds that
function as enzyme inhibitors, and particularly to a new class of
non-peptidic inhibitors of proteolytic enzymes.
[0004] 2. Related Art
[0005] Proteases are enzymes that cleave proteins at single,
specific peptide bonds. Proteases can be classified into four
generic classes: serine, thiol or cysteinyl, acid or aspartyl, and
metalloproteases (Cuypers et al., J. Biol. Chem. 257:7086 (1982)).
Proteases are essential to a variety of biological activities, such
as digestion, formation and dissolution of blood clots,
reproduction and the immune reaction to foreign cells and
organisms. Aberrant proteolysis is associated with a number of
disease states in man and other mammals. The human neutrophil
proteases, elastase and cathepsin G, have been implicated as
contributing to disease states marked by tissue destruction. These
disease states include emphysema, rheumatoid arthritis, corneal
ulcers and glomerular nephritis. (Barret, in Enzyme Inhibitors as
Drugs, Sandler, ed., University Park Press, Baltimore, (1980)).
Additional proteases such as plasmin, C-1 esterase, C-3 convertase,
urokinase, plasminogen activator, acrosin, and kallikreins play key
roles in normal biological functions of mammals. In many instances,
it is beneficial to disrupt the function of one or more proteolytic
enzymes in the course of therapeutically treating a mammal.
[0006] Serine proteases include such enzymes as elastase (human
leukocyte), cathepsin G, plasmin, C-1 esterase, C-3 convertase,
urokinase, plasminogen activator, acrosin, chymotrypsin, trypsin,
thrombin, factor Xa and kallikreins.
[0007] Human leukocyte elastase is released by polymorphonuclear
leukocytes at sites of inflammation and thus is a contributing
cause for a number of disease states. Cathepsin G is another human
neutrophil serine protease. Compounds with the ability to inhibit
the activity of these enzymes are expected to have an
anti-inflammatory effect useful in the treatment of gout,
rheumatoid arthritis and other inflammatory diseases, and in the
treatment of emphysema. Chymotrypsin and trypsin are digestive
enzymes. Inhibitors of these enzymes are useful in treating
pancreatitis. Inhibitors ofurokinase and plasminogen activator are
useful in treating excessive cell growth disease states, such as
benign prostatic hypertrophy, prostatic carcinoma and
psoriasis.
[0008] The serine protease thrombin occupies a central role in
hemostasis and thrombosis, and as a multifactorial protein, induces
a number of effects on platelets, endothelial cells, smooth muscle
cells, leukocytes, the heart and neurons (Tapparelli et al., Trends
in Pharmacological Sciences 14:366-376 (1993); Lefkovits and Topol,
Circulation 90(3): 1522-1536 (1994); Harker, Blood Coagulation and
Fibrinolysis 5 (Suppl 1):S47-S58 (1994)). Activation of the
coagulation cascade through either the intrinsic pathway (contact
activation) or the extrinsic pathway (activation by exposure of
plasma to a non-endothelial surface, damage to vessel walls or
tissue factor release) leads to a series of biochemical events that
converge on thrombin. Thrombin cleaves fibrinogen ultimately
leading to a hemostatic plug (clot formation), potently activates
platelets through a unique proteolytic cleavage of the cell surface
thrombin receptor (Coughlin, Seminars in Hematology 31(4):270-277
(1994)). and autoamplifies its own production through a feedback
mechanism. Thus, inhibitors of thrombin function have therapeutic
potential in a host of cardiovascular and non-cardiovascular
diseases, including: myocardial infarction; unstable angina;
stroke; restenosis; deep vein thrombosis; disseminated
intravascular coagulation caused by trauma, sepsis or tumor
metastasis; hemodialysis; cardiopulmonary bypass surgery; adult
respiratory distress syndrome; endotoxic shock; rheumatoid
arthritis; ulcerative colitis; induration; metastasis;
hypercoagulability during chemotherapy; Alzheimers disease; Down's
syndrome; fibrin formation in the eye; and wound healing. Other
uses include the use of said thrombin inhibitors as anticoagulants
either embedded in or physically linked to materials used in the
manufacture of devices used in blood collection, blood circulation,
and blood storage, such as catheters, blood dialysis machines,
blood collection syringes and tubes, blood lines and stents.
[0009] Factor Xa is another serine protease in the coagulation
pathway. Factor Xa associates with factor Va and calcium on a
phospholipid membrane thereby forming a prothrombinase complex.
This prothrombinase complex then converts prothrombin to thrombin
(Claeson, Blood Coagulation and Fibrinolysis 5:411-436 (1994);
Harker, Blood Coagulation and Fibrinolysis 5 (Suppl 1):S47-S58
(1994)). Inhibitors of factor Xa are thought to offer an advantage
over agents that directly inhibit thrombin since direct thrombin
inhibitors still permit significant new thrombin generation
(Lefkovits and Topol, Circulation 90(3):1522-1536 (1994); Harker,
Blood Coagulation and Fibrinolysis 5 (Suppl 1):S47-S58 (1994)).
[0010] A need continues to exist for non-peptidic compounds that
are potent and selective protease inhibitors, and which possess
greater bioavailability and fewer side-effects than currently
available protease inhibitors. Accordingly, new classes of potent
protease inhibitors, characterized by potent inhibitory capacity
and low mammalian toxicity, are potentially valuable therapeutic
agents for a variety of conditions, including treatment of a number
of mammalian proteolytic disease states.
SUMMARY OF THE INVENTION
[0011] The present invention is directed to novel compounds having
Formula I (below). Also provided are processes for preparing
compounds of Formula L The novel compounds of the present invention
are potent inhibitors of proteases, especially trypsin-like serine
proteases, such as chymotrypsin, trypsin, thrombin, plasmin and
factor Xa. Certain of the compounds exhibit antithrombotic activity
via direct, selective inhibition ofthrombin, or are intermediates
useful for forming compounds having antithrombotic activity.
[0012] The invention includes a composition for inhibiting loss
ofblood platelets, inhibiting formation of blood platelet
aggregates, inhibiting formation of fibrin, inhibiting thrombus
formation, and inhibiting embolus formation in a mammal, comprising
a compound ofthe invention in a pharmaceutically acceptable
carrier. These compositions may optionally include anticoagulants,
antiplatelet agents, and thrombolytic agents. The compositions can
be added to blood, blood products, or mammalian organs in order to
effect the desired inhibitions.
[0013] Also provided are methods of inhibiting or treating aberrant
proteolysis in a mammal, and methods for treating myocardial
infarction; unstable angina; stroke; restenosis; deep vein
thrombosis; disseminated intravascular coagulation caused by
trauma, sepsis or tumor metastasis; hemodialysis; cardiopulmonary
bypass surgery; adult respiratory distress syndrome; endotoxic
shock; rheumatoid arthritis; ulcerative colitis; induration;
metastasis; hypercoagulability during chemotherapy; Alzheimer's
disease; Downs syndrome; fibrin formation in the eye, and wound
healing. Other uses of compounds of the invention are as
anticoagulants either embedded in or physically linked to materials
used in the manufacture of devices used in blood collection, blood
circulation. and blood storage, such as catheters, blood dialysis
machines, blood collection syringes and tubes, blood lines and
stents.
[0014] The invention also includes a method for reducing the
thrombogenicity of a surface in a mammal by attaching to the
surface, either covalently or noncovalently, a compound of the
invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0015] The present invention is directed to a novel class of
benzamide and sulfonamide derivatives having Formula I: 2
[0016] or a solvate, hydrate or pharmaceutically acceptable salt
thereof, wherein:
[0017] L represents --C(O) or --SO.sub.2--; 3
[0018] or R.sup.1 and R.sup.2 can be taken together with the
nitrogen atom to which they are attached to form a three to seven
membered ring, either of which contains an additional nitrogen or
oxygen atom, and which is optionally benzo- or pyrido-fused, said
ring being preferably saturated, and said ring having one or two
optional substituents on either a ring carbon or nitrogen selected
from the group consisting of halogen, hydroxy, acyloxy, alkoxy,
aryloxy, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, heteroar(C.sub.1-4)alkyl, carboxyalkyl,
alkoxycarbonylalkyl, hydroxyalkoxyalkyl, cyano(C.sub.2-10)alkyl,
hydroxy(C.sub.2-10)alkyl, alkoxy(C.sub.2-10)alkyl, alkoxyalkyl,
mono- and di-alkylamino(C.sub.2-10)alkyl, carboxy, alkoxycarbonyl,
carboxamido, formyl, alkanoyl, aroyl, aralkanoyl, sulfonyl,
alkylsulfonyl, alkoxysulfonyl, and NR.sup.13R.sup.14 (when
C-substituted);
[0019] R.sup.12 represents hydrogen, C.sub.3-7 cycloalkyl,
C.sub.3-7 cycloalkenyl, C.sub.3-7 heterocycloalkyl, C.sub.3-7
heterocyFloalkenyl, aryl, or heteroaryl, which groups are
optionally substituted with C.sub.1-6 alkyl or hydroxy, or R.sup.12
represents diarylmethyl, diheteroarylmethyl, dicycloalkylmethyl or
(aryl)(heteroaryl)CH--;
[0020] Z and Z' independently represent a bond, a C.sub.1-6 alkyl
chain, a C.sub.3-6 alkenyl chain, or a C.sub.3-6 alkynyl chain,
where one or two nitrogen, oxygen, or sulfur atoms may be
optionally contained within each chain, and the chains are
optionally substituted by one or more groups selected from halogen,
hydroxy, CN, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6
alkoxy(C.sub.1-6)alkyl, C.sub.1-6 acyloxy, NR.sup.13R.sup.14,
NHCOR.sup.15, NHSO.sub.2R.sup.16, COR.sup.15, CO.sub.2R.sup.15,
CONR.sup.13R.sup.14, and SO.sub.2NR.sup.17R.sup.18;
[0021] provided that when one of R.sup.1 or R.sup.2 is C.sub.3-8
alkyl, cycloalkyl, C.sub.3-8 alkenyl, C.sub.3-8 alkynyl, aryl,
aralkyl, or heteroaryl, any of which is optionally substituted,
then the other of R.sup.1 or R.sup.2 is other than hydrogen, alkyl,
aralkyl, aryl, hydroxy(C.sub.2-10)alkyl, amino(C.sub.2-10)alkyl,
monoalkylamino(C.sub.2-- 10)alkyl, dialkylamino(C.sub.2-10)alkyl or
carboxyalkyl;
[0022] R.sup.13-R.sup.16 represent hydrogen, C.sub.1-6 alkyl,
C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl,
C.sub.6-10 aryl, mono- or di-hydroxy(C.sub.6-10)aryl, C.sub.6-10
ar(C.sub.1-4)alkyl, pyridyl, pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)-alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono- and
di-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl;
[0023] or R.sup.3 and R.sup.14 form a C.sub.3-7 heterocycloalkyl
ring,
[0024] or R.sup.16 additionally may represent trifluoromethyl;
[0025] R.sup.17 and R.sup.18 are independently selected from the
group consisting of hydrogen, C.sub.1-6 alkyl, C.sub.3-7
cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.6-10 aryl,
C.sub.6-10 ar(C .sub.1-4)alkyl, pyrdyl pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)alkyl, C.sub.1-4 alkoxycarbonyl-(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, and mono- and
di-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl,
[0026] or R.sup.17 and R.sup.18 can be taken together with the
nitrogen atom to which they are attached to form a heterocyclic
ring selected from the group consisting of N-morpholinosulfonyl,
N-piperazinylsulfonyl (optionally N' substituted with C.sub.1-6
alkyl, C.sub.1-6 hydroxyalkyl, C.sub.6-10 aryl, C.sub.6-10
aryl(C.sub.1-6)alkyl, C.sub.1-6 alkylsulfonyl, C.sub.6-10
arylsulfonyl, C.sub.1-6 alkylcarbonyl, morpholino or C.sub.6-10
arylcarbonyl), N-pyrrolylsulfonyl, N-piperidinylsulfonyl,
N-pyrrolidinylsulfonyl, N-dihydropyridylsulfonyl,
N-indolylsulfonyl, wherein said heterocyclic ring can be optionally
C-substituted;
[0027] R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are each independently
one of hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, optionally
substituted aryl, optionally substituted aralkyl, optionally
substituted heteroaryl, trifluoromethyl, halogen, hydroxyalkyl,
cyano, nitro, carboxamido, --CO.sub.2R.sup.x, --CH.sub.2OR.sup.x or
--OR.sup.x, or when present on adjacent carbon atoms, R.sup.3 and
R.sup.4 may also be taken together to form one of --CH=CH--CH=CH--
or --(CH.sub.2).sub.q-- where q is from 2 to 6, and R.sup.5 and
R.sup.6 are defined as above;
[0028] R.sup.x, in each instance, is independently one of hydrogen,
alkyl or cycloalkyl wherein said alkyl or cycloalkyl groups may
optionally have one or more unsaturations;
[0029] Y is one of --O--, --NR.sup.19--, --S--, --CHR.sup.19-- or a
covalent bond;
[0030] R.sup.19, in each instance, is independently hydrogen,
C.sub.1-6 alkyl, benzyl, phenyl, C.sub.2-10 hydroxyalkyl,
C.sub.2-10 aminoalkyl, C.sub.1-4 monoalkylamino(C.sub.2-8)alkyl,
C.sub.1-4 dialkylamino(C.sub.2-8)alkyl or C.sub.2-10
carboxyalkyl;
[0031] R.sup.7 is one of hydrogen, alkyl, aralkyl, aryl,
hydroxyalkyl, aminoalkyl, monoalkylarninoalkyl, dialkylaminoalkyl,
carboxyalkyl, hydroxy, alkoxy, aralkoxy, aryloxy, heteroaryloxy, or
mono- or di- alkylamino, provided that n is other than zero when
R.sup.7 is hydroxy, alkoxy, aralkoxy, aryloxy, heteroaryloxy, or
mono- or di- alkylamino;
[0032] R.sup.8, R.sup.9 and R.sup.10 are each independently one of
hydrogen, alkyl. aralkyl, aryl, hydroxyalkyl, aminoalkyl,
monoalkylaminoalkyl, dialkylaminoalkyl or carboxyalkyl;
[0033] or R.sup.7 and R.sup.8 are taken together to form
--(CH.sub.2).sub.y-- where y is zero (a bond), 1 or 2, while
R.sup.9 and R.sup.10 are defined as above; or R.sup.7 and R.sup.10
are taken together to form --(CH.sub.2).sub.t--, where t is zero (a
bond), or 1 to 8, while R.sup.8 and R.sup.9 are defined as above;
or R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r--, where r is 2-8, while R.sup.7 and R.sup.10
are defined as above;
[0034] X is oxygen or NH;
[0035] R.sup.11 is one of hydrogen, alkyl, aralkyl, aryl,
hydroxyalkyl, aminoalkyl, monoalkylamino(C.sub.2-10)alkyl,
dialkylamino(C.sub.2-10)alky- l or carboxyalkyl;
[0036] R.sup.a, R.sup.b and R.sup.c are independently hydrogen,
alkyl, hydroxy, alkoxy, aryloxy, aralkoxy, alkoxycarbonyloxy, cyano
or --CO.sub.2R.sup.w;
[0037] R.sup.w is alkyl, trichloroethyl, cycloalkyl, phenyl,
benzyl, 4
[0038] where R.sup.d and R.sup.e are independently hydrogen,
C.sub.1-6alkyl, C.sub.2-6 alkenyl or phenyl, R.sup.f is hydrogen,
C.sub.1-6alkyl, C.sub.2-6 alkenyl or phenyl, R.sup.g is hydrogen,
C.sub.1-6 alky C.sub.2-6 alkenyl or phenyl, and R.sup.h is aralkyl
or C.sub.1-6 alkyl;
[0039] n is from zero to 8; and
[0040] m is from zero to 4.
[0041] The moiety --L--NR.sup.1R.sup.2 is attached to the benzene
ring in a position ortho-, meta-, or para- to Y, with the meta-
position being preferred.
[0042] Preferably, the compounds have the structure of Formula Ia:
5
[0043] wherein each of the groups is as defined for Formula I
above.
[0044] Referring to the general Formula I and Formula Ia, where
R.sup.2 represents a group 6
[0045] Z' is suitably C.sub.3-6 alkenyl, e.g., allyl, or C.sub.1-6
alkyl, e.g., methyl, ethyl, propyl or pentyl, which optionally
contains an oxygen group within the chain and is optionally
substituted by a group selected from hydroxy, C.sub.1-6 alkoxy,
NHSO.sub.2R.sup.16, CO.sub.2R.sup.15, CONR.sup.13R.sup.14, or
SO.sub.2NR.sup.17R.sup.18, and R.sup.12 is suitably hydrogen,
C.sub.3-7 heterocycloalkyl, e.g., pyrrolidine or morpholine, aryl,
e.g., phenyl which is optionally substituted by CO.sub.2R.sup.15,
or heteroaryl, e.g., oxadiazole optionally substituted by hydroxy,
triazole, or tetrazole optionally substituted by C.sub.1-6
alkyl.
[0046] Referring to the general Formula I and Formula Ia where
R.sup.1 represents a group 7
[0047] Z is suitably a bond or C.sub.1-6 alkyl group, e.g., methyl,
isopropyl or isobutyl, and R.sup.12 suitably represents hydrogen,
C.sub.3-7 cycloalkyl, aryl, or heteroarnI, When Z represents a
bond, R.sup.12 is preferably optionally substituted phenyl,
C.sub.3-7 cycloalkyl, e.g., cyclobutyl, cyclopentyl or cyclohexyl,
diphenylmethyl or dicyclohexylmethyl. When Z represents a C.sub.1-4
alkyl group, R.sup.12 is preferably hydrogen, cycloalkyl, e.g.,
cyclohexyl, or heteroaryl, e.g., thienyl or furyl.
[0048] Useful values of R.sup.12 include C.sub.6-10 aryl,
pyridinyl, thiophenyl (i.e., thiophene), quinazolinyl, quinolinyl,
isoquinolinyl, or tetrahydroquinolinyl, any of which is optionally
substituted by one or two of hydroxy, nitro, trifluoromethyl,
halogen, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, C.sub.1-6 aminoalkyl,
C.sub.1-6 aminoalkoxy, amino, mono(C.sub.1-4)alkylamino,
di(C.sub.1-4)alkylamino, C.sub.2-6 alkoxycarbonylamino, C.sub.2-6
alkoxycarbonyl, carboxy, C.sub.1-6 hydroxyalkyl, C.sub.2-6
hydroxyalkoxy, C.sub.2-10 mono(carboxyalkyl)amino- , bis(C.sub.2-10
carboxyalkyl)amino, C.sub.6-14 ar(C.sub.1-6) alkoxycarbonyl,
C.sub.2-6 alkynylcarbonyl, C.sub.1-6 alkylsulfonyl, C.sub.2-6
alkenylsulfonyl, C.sub.2-6 alkynylsulfonyl, C.sub.1-6
alkylsulfinyl, C.sub.1-6 alkylsulfonamido, amidino, guanidino,
C.sub.1-6 alkyliminoamino, formyliminoamino, C.sub.2-6
carboxyalkoxy, C.sub.2-6 carboxyalkyl, carboxyalkylamino, cyano,
trifluoromethoxy, perfluoroethoxy, C.sub.1-6 acyloxy, and
R.sup.17R.sup.18NSO.sub.2--,
[0049] where R.sup.17 and R.sup.18 are independently selected from
the group consisting of hydrogen, C.sub.1-6 alkyl, C.sub.3-7
cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6 alkynyl, C.sub.6-10 aryl,
C.sub.6-10ar(C.sub.1-4)alkyl, pyridyl, pyridyl(C.sub.1-4)alkyl,
carboxy(C.sub.1-6)alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono- and
di-(C.sub.1-4)alkylamino(C.sub.2-6)alky- l, or R.sup.17 and
R.sup.18 can be taken together with the nitrogen atom to which they
are attached to form a heterocyclic ring selected from the group
consisting of N-morpholinosulfonyl, N-piperazinylsulfonyl
(optionally N' substituted with C.sub.1-6 alkyl, C.sub.1-6
hydroxyalkyl, C.sub.6-10 aryl, C.sub.6-10 aryl(C.sub.1-6)alkyl,
C.sub.1-6 alkylsulfonyl, C.sub.6-10 arylsulfonyl, C.sub.1-6
alkylcarbonyl, morpholino or C.sub.6-10 arylcarbonyl),
N-pyrrolylsulfonyl, N-piperidinylsulfonyl, N-pyrrolidinylsulfonyl,
N-dihydropyridylsulfonyl, N-indolylsulfonyl, wherein said
heterocyclic ring can be optionally substituted with one or two of
hydroxy, C.sub.1-8 alkanoyloxy, C.sub.1-6 alkoxy, C.sub.6-10
aryloxy, amino, mono- and di- C.sub.1-6 alkylamino, C.sub.1-8
alkanoylamino, C.sub.1-4 alkyl, C.sub.3-7 cycloalkyl, C.sub.6-10
aryl, C.sub.6-10 ar(C.sub.1-4)alkyl, heterocycle heterocycloalkyl,
carboxy(C.sub.1-6)alkyl, C.sub.1-4 alkoxycarbonyl(C.sub.1-4)alkyl,
cyano(C.sub.2-6)alkyl, hydroxy(C.sub.2-6)alkyl, C.sub.1-4
alkoxy(C.sub.2-6)alkyl, mono- and
di-(C.sub.1-4)alkylamino(C.sub.2-6)alkyl, carboxy, C.sub.1-6
alkoxycarbonyl, carboxamido, formyl, C.sub.1-6 alkanoyl, C.sub.6-10
aroyl, C.sub.6-10 ar(C.sub.1-4)alkanoyl, sulfonyl, C.sub.1-6
alkylsulfonyl, C.sub.1-6 alkoxysulfonyl, sulfonamido, phosphonyl,
phosphoramido, or phosphinyl.
[0050] R.sup.12 is more preferably one of phenyl, naphthyl,
pyridyl, thiophenyl, quinolinyl or isoquinolinyl, optionally
substituted by one or two of chloro, methoxy, methyl,
trifluoromethyl, phenyl, cyano, nitro, amino, dimethylamino,
alkylsulfonyl, arylsulfonyl, or R.sup.17R.sup.18NSO.sub.2--, where
R.sup.17 and R.sup.18 are defined as above.
[0051] Particularly preferred combinations of R.sup.1 and R.sup.2
include:
[0052] (A) R.sup.1 and R.sup.2 are taken together with the nitrogen
to which they are attached to form a C.sub.3-7 heterocycloalkyl or
C.sub.3-7 heterocycloalkenyl group, optionally benzo fused and
optionally including an oxygen atom or an additional nitrogen atom,
and which may be optionally substituted by C.sub.1-6 alkyl,
hydroxy, C.sub.1-4 alkoxy, C.sub.2-6 alkoxycarbonyl, formyl,
(C.sub.6-10)ar(C.sub.1-4)alkyl, C.sub.6-10 aryl, pyridyl,
hydroxyalkoxyalkyl, halogen, or NR.sup.13R.sup.14; or
[0053] (B) R.sup.1 is C.sub.3-7 cycloalkyl or C.sub.3-7
cycloalkenyl, either of which is optionally substituted by
C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy, halogen, carboxylic
acid. a C.sub.1-4 carboxylic acid ester group, or
NR.sup.13R.sup.14, and R.sup.2 is C.sub.3-6 alkenyl, or C.sub.3-6
alkynyl, either of which is optionally substituted by C.sub.1-6
alkyl, hydroxy, C.sub.1-4 alkoxy, halogen, carboxylic acid, a
C.sub.1-4 carboxylic acid ester group, or NR.sup.13R.sup.14; or
[0054] (C) R.sup.1 is C.sub.3-7 heterocycloalkyl(C.sub.1-6)alkyl,
C.sub.3-7 heterocycloalkenyl(C.sub.1-6)alkyl,
heteroaryl(C.sub.1-6)alkyl, C.sub.3-7
heterocycloalkyl(C.sub.3-6)alkenyl, C.sub.3-7
heterocycloalkenyl(C.sub.3-6)alkenyl, heteroaryl(C.sub.3-6)alkenyl,
C.sub.3-7 heterocycloalkyl(C.sub.3-6)alkynyl. C.sub.3-7
heterocycloalkenyl(C.sub.3-6)alkynyl, heteroaryl(C.sub.3-6)alkynyl,
di(C.sub.5-10aryl)(C.sub.1-3)alkyl, di(C.sub.3-8
cycloalkyl)(C.sub.1-3)al- kyl or di(C.sub.3-8
cycloalkenyl)(C.sub.1-3)alkyl, any of which is optionally
substituted by C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy, halogen,
carboxylic acid, a C.sub.1-4 carboxylic acid ester group, or
NR.sup.13R.sup.14; and
[0055] R.sup.2 is a group 8
[0056] where R.sup.12 and Z' have the values and preferred values
defined above.
[0057] R.sup.3 can represent hydrogen, Cl-.sub.3 alkyl, halogen, or
C.sub.1-2 alkoxy. R.sup.3 is preferably C.sub.1-3 alkyl, e.g.,
methyl, or halogen, e.g., chlorine or bromine.
[0058] R.sup.4, R.sup.5 and R.sup.6 can independently represent
hydrogen, or halogen. R.sup.4, R.sup.5 and R.sup.6 are preferably
hydrogen, or halogen, e.g., fluorine.
[0059] Preferred values of Y are divalent oxygen (--O--),
--NR.sup.9-- or a covalent bond, most preferably --O--.
[0060] Preferred values of R.sup.19 are hydrogen, C.sub.1-6 alkyl
and C.sub.3-6 cycloalkyl.
[0061] Preferred values of R.sup.11 are hydrogen, C.sub.1-6 alkyl,
or C.sub.6-10 ar(C.sub.1-6)alkyl.
[0062] Preferred values of R.sup.7, R.sup.8, R.sup.9 and R.sup.10
are independently one of hydrogen, C.sub.1-6 alkyl C.sub.6-10
ar(C.sub.1-6)alkyl, C.sub.6-10 aryl, C.sub.2-10 hydroxyalkyl or
C.sub.2-7 carboxyalkyl. Useful values of R.sup.7, R.sup.8, R.sup.9
and R.sup.10 include hydrogen, methyl, ethyl, propyl, n-butyl,
benzyl, phenylethyl, 2-hydroxyethyl, 3-hydroxypropyl,
4-hydroxybutyl, 2-carboxymethyl, 3-carboxyethyl and
4-carboxypropyl. Additional preferred compounds are those wherein
R.sup.7 and R.sup.8 are taken together to form --(CH.sub.2).sub.y--
where y is most preferably 2. Another group of preferred compounds
are those where R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r-- where r is most preferably 2.
[0063] A preferred value of X is O.
[0064] Preferred values of R.sup.a, R.sup.b and R.sup.c in Formula
I are hydrogen, hydroxy, C.sub.1-6 alkyl, C.sub.1-6 alkoxy, cyano
or --CO.sub.2R.sup.w, where R.sup.w, in each instance, is
preferably one of C.sub.1-4alkyl, C.sub.4-7cycloalkyl or
benzyloxycarbonyl. Suitable values of R.sup.a, R.sup.b and R.sup.c
include hydrogen, methyl, ethyl, propyl, n-butyl, hydroxy, methoxy,
ethoxy, cyano, --CO.sub.2CH.sub.3, --CO.sub.2CH.sub.2CH.sub.3 and
--CO.sub.2CH.sub.2CH.sub.2CH.sub.3. In the most preferred
embodiments, R.sup.a, R.sup.b and R.sup.c are each hydrogen.
[0065] Also preferred at R.sup.a, R.sup.b and R.sup.c is the group
--CO.sub.2R.sup.w, where R.sup.w is one of 9
[0066] where R.sup.d-R.sup.h are defined as above. When R.sup.a,
R.sup.b and R.sup.c are --CO.sub.2R.sup.w, where R.sup.w is one of
one of these moieties, the resulting compounds are prodrugs that
possess desirable formulation and bioavailability characteristics.
A preferred value for each of R.sup.d, R.sup.e and R.sup.g is
hydrogen, R.sup.f is methyl, and preferred values for R.sup.h
include benzyl and tert-butyl.
[0067] Preferred values of n in Formula I include from zero to 6,
more preferably from zero to 4, and most preferably zero, 1 or 2.
Preferred values of m include from zero to 4, more preferably zero,
1, 2 or 3.
[0068] A preferred group of compounds falling within the scope of
the present invention include compounds of Formula Ia wherein:
[0069] (A) R.sup.1 and R.sup.2 are taken together with the nitrogen
to which they are attached to form a C.sub.3-7 heterocycloalkyl or
C.sub.3-7 heterocycloalkenyl group, optionally benzo fused and
optionally including an oxygen atom or an additional nitrogen atom,
and which may be optionally substituted by C.sub.1-6 alkyl,
hydroxy, C.sub.1-4 alkoxy, C.sub.2-6 alkoxycarbonyl, formyl,
(C.sub.6-10)ar(C.sub.1-4)alkyl, C.sub.6-10 aryl, pyridyl,
hydroxyalkoxyalkyl, halogen, or NR.sup.13R.sup.14; or
[0070] (B) R.sup.1 is C.sub.3-7 cycloalkyl or C.sub.3-7
cycloalkenyl, either of which is optionally substituted by
C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy, halogen, carboxylic
acid, a C.sub.1-4 carboxylic acid ester group, or
NR.sup.13R.sup.14, and R.sup.2 is C.sub.3-6 alkenyl, or C.sub.3-6
alkynyl, either of which is optionally substituted by C.sub.1-6
alkyl, hydroxy. C.sub.1-4 alkoxy, halogen, carboxylic acid, a
C.sub.1-4 carboxylic acid ester group, or NR.sup.13R.sup.14; or
[0071] (C) R.sup.1 is C.sub.3-7 heterocycloalkyl(C.sub.1-6)alkyl,
C.sub.3-7 heterocycloalkenyl(C.sub.1-6)alkyl,
heteroaryl(C.sub.1-6)alkyl, C.sub.3-7
heterocycloalkyl(C.sub.3-6)alkenyl, C.sub.3-7
heterocycloalkenyl(C.sub.3-6)alkenyl, heteroaryl(C.sub.3-6)alkenyl,
C.sub.3-7 heterocycloalkyI(C.sub.3-6)alkynyl, C.sub.3-7
heterocycloalkenyl(C.sub.3-6)alkynyl, heteroaryl(C.sub.3-6)alkynyl,
di(C.sub.5-10 aryl)(C.sub.1-3)alkyl, di(C.sub.3-8
cycloalkyl)(C.sub.1-3)a- lkyl or di(C.sub.3-8
cycloalkenyl)(C.sub.1-3)alkyl, any of which is optionally
substituted by C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy, halogen,
carboxylic acid, a C.sub.1-4 carboxylic acid ester group, or
NR.sup.13R.sup.14; and
[0072] R.sup.2 is a group 10
[0073] where R.sup.12 and Z' have the values and preferred values
defined above;
[0074] R.sup.13 and R.sup.14 independently represent hydrogen,
C.sub.1-6 alkyl, C.sub.3-7 cycloalkyl, C.sub.2-6 alkenyl, C.sub.2-6
alkynyl, C.sub.6-10 aryl, C.sub.6-10 ar(C.sub.1-4)alkyl, pyridyl,
pyridyl(C.sub.1-4)alkyl, carboxy(C.sub.1-6)-alkyl, C.sub.1-4
alkoxycarbonyl(C.sub.1-4)alkyl, cyano(C.sub.2-6)alkyl,
hydroxy(C.sub.2-6)alkyl, C.sub.1-4 alkoxy(C.sub.2-6)alkyl, mono-
and di- (C.sub.1-4)alkylamino(C.sub.2-6)alkyl;
[0075] or R.sup.13 and R.sup.14 form a C.sub.3-7 heterocycloalkyl
ring;
[0076] R.sup.3 is hydrogen, C.sub.1-3 alkyl, halogen or C.sub.1-2
alkoxy;
[0077] R.sup.4, R.sup.5 and R.sup.6 are hydrogen or halogen;
[0078] Y is --O--;
[0079] R.sup.a, R.sup.b and R.sup.c are each one of hydrogen,
C.sub.1-4 alkyl, hydroxy, C.sub.1-4 alkoxy. phenoxy, C.sub.1-4
alkyloxycarbonyl, benzyloxycarbonyl, cyano, 11
[0080] where R.sup.h is benzyl, methyl, ethyl, isopropyl, sec-butyl
or t-butyl, and where R.sup.f is hydrogen or C.sub.1-6 alkyl
[0081] R.sup.11 is one of hydrogen, C.sub.1-6 alkyl C.sub.6-10
ar(C.sub.1-6)alkyl, C.sub.6-10 aryl, C.sub.2-10 hydroxyalkyl,
C.sub.2-10 aminoalkyl, mono(C.sub.1-4)alkylamino(C.sub.2-8)alkyl,
di(C.sub.1-4)alkylamino(C.sub.2-8)alkyl or C.sub.2-10
carboxyalkyl;
[0082] R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are independently one
of hydrogen, C.sub.1-6 alkyl, C.sub.2-10 carboxyalkyl or C.sub.2-10
hydroxyalkyl, or R.sup.7 and R.sup.8 are taken together to form
--CH.sub.2).sub.y-- where y is zero, 1 or 2, while R.sup.9 and
R.sup.10 are defined as above; or R.sup.7 and R.sup.10 are taken
together to form --(CH.sub.2).sub.t--, where t is zero (a bond), or
1, 2 or 3, while R.sup.8 and R.sup.9 are defined as above; or
R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r--, where r is 2, 3, or 4, while R.sup.7 and
R.sup.10 are defined as above;
[0083] R.sup.20 is hydrogen, or C.sub.1-10 alkyl, optionally
substituted with amino, mono(C.sub.1-4)alkylamino, C.sub.1-6
alkoxy, hydroxy, carboxy, phenyl, C.sub.1-4 alkyloxycarbonyl,
C.sub.6-10 ( ar(C.sub.1-4)alkoxycarbonyl, C.sub.1-6 acylamino,
cyano or trifluoromethyl;
[0084] n is from zero to 4; and m is from zero to 4.
[0085] An especially preferred group of compounds include compounds
of Formula Ia wherein:
[0086] R.sup.1 is cyclopentyl cyclohexyl or cycloheptyl; R.sup.2 is
aftyl, diphenylmethyl or dicyclohexylmethyl;
[0087] R.sup.3 is hydrogen, methyl, chloro or C.sub.1-C.sub.2
alkoxy;
[0088] R.sup.4, R.sup.5 and R.sup.6 are hydrogen or halogen;
[0089] Y is --O--;
[0090] R.sup.a, R.sup.b and R.sup.c are hydrogen, hydroxy, 12
[0091] where R.sup.h is benzyl or t-butyl, and where R.sup.f is
hydrogen or methyl;
[0092] R.sup.11 is hydrogen, C.sub.1-4 alkyl, C.sub.2-4
hydroxyalkyl, C.sub.2-4 carboxyalkyl, C.sub.2-4 aminoalkyl,
dimethylamino(C.sub.2-8)alk- yl, or
methylamino(C.sub.2-8)alkyl;
[0093] R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are independently one
of hydrogen, C.sub.1-6 alkyl, C.sub.2-10, hydroxyalkyl or
C.sub.2-10 carboxyalkyl, or R.sup.7 and R.sup.8 are taken together
to form --(CH.sub.2).sub.y-- where y is zero, 1 or 2, while R.sup.9
and R.sup.10 are defined as above; or R.sup.7 and R.sup.10 are
taken together to form --(CH.sub.2).sub.q--, where q is zero (a
bond), or 1, 2 or 3, while R.sup.8 and R.sup.9 are defined as
above; or R.sup.8 and R.sup.9 are taken together to form
--(CH.sub.2).sub.r--, where r is 2, 3 or 4, while R.sup.7 and
R.sup.10 are defined as above;
[0094] X is --O--;
[0095] n is from zero to 4; and
[0096] m is zero, 1, 2 or 3.
[0097] An especially preferred subclass ofthe compounds of
FormulaIis defined by compounds of Formula IIa and IIb: 13
[0098] or a solvate, hydrate, pharmaceutically acceptable salt or
prodrug thereof, wherein:
[0099] R.sup.1A represents a group: 14
[0100] wherein Z.sup.A represents a bond or C.sub.1-6 alkyl; and
R.sup.12A represents hydrogen, C.sub.3-7- cycloalkyl, C.sub.1-6
alkoxy, aryl optionally substituted by halogen, hydroxy,
heteroaryl, diphenylmethyl or dicyclohexylmethyl;
[0101] R.sup.2A represents a group: 15
[0102] wherein Z.sup.B represents C.sub.3-6 alkenyl or C.sub.1-6
alkyl optionally substituted by CO.sub.2R.sup.15 or COR.sup.15;
R.sup.12B represents hydrogen, C.sup.1-6 alkoxy, mono- or di-
C.sub.1-3 alkylamino, phenyl substituted by CO.sub.2R.sup.15,
oxadiazole substituted by a hydroxy group, oranunsubstituted
C-linkedtetrazole group; and R.sup.15 is C.sub.1-3 alkyl or mono-
or di-hydroxyphenyl;
[0103] or R.sup.1A and R.sup.2A can be taken together with the
nitrogen atom to which they are attached to form a three to seven
membered ring, either of which contains an additional nitrogen or
oxygen atom, and which is optionally benzo or pyrido fused, said
ring being preferably saturated, and said ring having one or two
optional substituents on either a ring carbon or nitrogen selected
from the group consisting of halogen, hydroxy, acyloxy, alkoxy,
aryloxy, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, aralkyl,
heteroaryl, heteroar(C.sub.1-4)alkyl, carboxyalkyl,
alkoxycarbonylalkyl, hydroxyalkoxyalkyl, cyano(C.sub.2-10)alkyl,
hydroxy(C.sub.2-10)alkyl, alkoxy(C.sub.2-10)alkyl- , alkoxyalkyl,
mono- and di-alkylamino(C.sub.2-10)alkyl, carboxy, alkoxycarbonyl,
carboxamido, formyl, alkanoyl, aroyl, aralkanoyl, sulfonyl,
alkylsulfonyl, alkoxysulfonyl, and NR.sup.13R.sup.14 (when
C-substituted):
[0104] R.sup.3A represents C.sub.1-3 alkyl or halogen, preferably
chloro, bromo or methyl;
[0105] R.sup.11A represents hydrogen, C.sub.6-10 ar(C.sub.1-4)alkyl
or C.sub.1-4 alkyl;
[0106] R.sup.a, R.sup.b and R.sup.c are hydrogen;
[0107] a is from zero to 8, preferably zero, 1, 2 or 3; and b is
from zero to 8, preferably 1, 2 or 3.
[0108] An even more especially preferred group of compounds include
compounds of Formula *Ia wherein:
[0109] R.sup.1A represents a group: 16
[0110] Z.sup.A represents a bond or C.sub.1-6 alkyl; and R.sup.12A
represents hydrogen, C.sub.3-7 cycloalkyl, C.sub.1-6 alkoxy, aryl
optionally substituted by halogen or hydroxy, or heteroaryl;
[0111] R.sup.2A represents a group: 17
[0112] wherein Z.sup.B represents C.sub.3-6 alkenyl or C.sub.1-6
alkyl optionally substituted by CO.sub.2R.sup.15 or COR.sup.15;
R.sup.12B represents hydrogen, C.sub.1-6 alkoxy, or mono- or di-
C.sub.1-3 alkylamino; and R.sup.15 is C.sub.1-3 alkyl or mono- or
di-hydroxyphenyl;
[0113] or R.sup.1A and R.sup.2A are taken together with the
nitrogen to which they are attached to form a C.sub.3-7
heterocycloalkyl or C.sub.3-7 heterocycloalkenyl group, optionally
benzo fused and optionally including an oxygen atom or an
additional 15 nitrogen atom, and which may be optionally
substituted by C.sub.1-6 alkyl, hydroxy, C.sub.1-4 alkoxy,
C.sub.2-6 alkoxycarbonyl, formyl, (C.sub.6-10)ar(C.sub.1-4)alkyl,
C.sub.6-10 aryl, pyridyl, hydroxy(C.sub.1-4)alkoxy(C.sub.1-4)alkyl,
halogen, or NR.sup.13R.sup.14, where R.sup.13 and R.sup.14 are as
defined above;
[0114] R.sup.3A represents halogen, preferably chloro;
[0115] X.sup.A is --O--;
[0116] R.sup.11A is hydrogen, C.sub.6-10 ar(C.sub.1-4)alkyl or
C.sub.1-4 alkyl;
[0117] R.sup.a, R.sup.b and R.sup.c are hydrogen; and
[0118] a is 1.
[0119] Non-limiting examples of compounds of the present invention
include [3-{5-chloro-3-(N-cyclopentyl-N-[prop-2-enyl]aminocarbonyl)
phenoxy }propoxyamino]carboxamidine hydrochloride, [3-{5-chloro-3
-(4-benzylpiperidinylcarbonyl) phenoxy}propoxyamino]carboxamidine
trifluoroacetate,
[3-{5-chloro-3-(N,N-bis[2-methoxyethyl]aminocarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(N-methyl-N-[2-{2-pyridyl}ethyl]aminocarbonyl)phenoxy3prop-
oxyamino]carboxamidinetrifluoroacetate,
[3-{5-chloro-3-(N-methyl-N-[3-5pyr- idylmethyl]aminocarboryl)
phenoxy }propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(N-ethyl-N-[4-pyridylmethyl]aminocarbony- l) phenoxy
}propoxyamino]carboxamidine trifluoroacetate, ethyl
2-[5-{3-(amidinoaminooxy)propoxy}-3-chlorophenyl]-N-
{2-pyridylmethyl}aminocarbonyl]acetate trifluoroacetate, methyl
2-[5-{3-(amidinoaminooxy)propoxy}-3-chlorophenyl]-N-{2-pyridylmethyl}amin-
ocarbonyl]acetate trifluoroacetate,
[3-{5-chloro-3-([2-{3,4-dihydroxypheny- l }-2 -oxoethyl]-N-methyl
aminocarbonyl)phenoxy}propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(N-[2-{dimethy lamino
}ethyl]-N-4methylaminocarbonyl)phenoxy}propoxyarnino]carboxamidine
trifluoroacetate, [3-{5-chloro-3-(4-formypiperazinycarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate, [3-{5-chloro-3
-(4-phenylpiperazinylcarbonyl) phenoxy}propoxyamino]carboxamidine
trifluoroacetate, [3-{5-chloro-3-(4-benzylpiperazinylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(N,N-dimethylaminocarbonyl) phenoxy
}propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(piperidinyl- carbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(4-[2-pyridyl]piperazinylcarbonyl)
phenoxy}propoxyamino]ca- rboxamidine trifluoroacetate, [3-
{5-chloro-3-(4-[4-pyridyl]piperazinylcar- bonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate, [3-{5-chloro-3
-(2-[1,2,3 ,4-tetrahydro]isoquinolinylcarbonyl) phenoxy
}propoxyamino]carboxamidine trifluoroacetate,
[3-{5-chloro-3-(azaperhydro- ep inylcarbonyl) phenoxy
}propoxyamino]carboxamidine trifluoroacetate, ethyl
3-({5-[3-(amidinoaminooxy)propoxy]-3-chlorophenyl}-N-benzylcarbonyl-
amino)propanoate trifluoroacetate, ethyl
1-({5-[3-(amidinoaminooxy)propoxy-
]-3-chlorophenyl}carbonyl)piperidine-4-carboxylate
trifluoroacetate, [3 - {5-chloro-3-(morpholin-4-ylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate, and methyl 2-(
{ 5-[3-(amidinoaminooxy)propoxy]-3-chlor-
ophenyl}-N-methylcarbonylamino)acetate trifluoroacetate.
[0120] Alternative embodiments of the present invention include
compounds of Formula I in which two "R" groups together form a
saturated or unsaturated hydrocarbon bridge, thus forming an
additional cyclic moiety in the resulting compounds. Alternative
embodiments include compounds of Formula I wherein Z,
R.sup.1-R.sup.4, Y, m and n are as defined above; and:
[0121] A. R.sup.7 and R.sup.10 are taken together to form
--(CH.sub.2).sub.o--, where o is 1, 2 or 3;
[0122] R.sup.9 is hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl or
carboxyalkyl;
[0123] R.sup.8 is hydrogen and R.sup.11, R.sup.a, R.sup.b and
R.sup.c are defined as above; or
[0124] B. R.sup.9 is hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl
or carboxyalkyl;
[0125] R.sup.7 is hydrogen;
[0126] R.sup.8 and R.sup.10 are taken together to form
--(CH.sub.2)--(CH.sub.2)--(CH.sub.2).sub.p--, where p is 1, 2 or 3;
and
[0127] R.sup.11, R.sup.a, R.sup.b and R.sup.c are defined as above;
or
[0128] C. R.sup.11 and R.sup.b are taken together to form
--(CH.sub.2)--(CH.sub.2).sub.r-- or .dbd.CH--N.dbd.CH--NH--, where
r is 1, 2 or 3;
[0129] R.sup.a is hydrogen or hydroxy;
[0130] R.sup.c is hydrogen, alkyl, hydroxy, alkoxy, aryloxy,
aralkoxy, alkoxycarbamoyloxy, cyano or --CO.sub.2R.sup.w--, where
R.sup.w is as defined above;
[0131] R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are each
independently one of hydrogen, alkyl, aralkyl, aryl, hydroxyalkyl
or carboxyalkyl, or R.sup.7 and R.sup.8 are taken together to form
--(CH.sub.2).sub.y--, where y is zero, 1 or 2; or
[0132] D. R.sup.a and R.sup.b are taken together to form
--CH.sub.2--(CH.sub.2).sub.s--, where s is 1 or2;
[0133] R.sup.11 is hydrogen, alkyl, alkoxy, aryloxy, aralkoxy,
alkoxycarbonyloxy, cyano or --CO.sub.2R.sup.w--, where R.sup.w is
as defined above; and
[0134] R.sup.7, R.sup.8, R.sup.9 and R.sup.10 are each
independently one of hydrogen, alkyl, 5 aralkyl, aryl, hydroxyalkyl
or carboxyalkyl, or R.sup.7 and R.sup.8 are taken together to form
--(CH.sub.2).sub.y--, where y is zero, 1 or 2.
[0135] Thus, compounds having Formulae III, IV, V and V
(representing embodiments A, B, C and D, respectively) are
contemplated: 18
[0136] wherein R.sup.1-R.sup.11, Z, Y, R.sup.a-R.sup.c, n, m, o, p,
r and s are defined as above. Preferred values for each of these
variables are the same as described for Formula I.
[0137] It is also to be understood that the present invention is
considered to include stereoisomers as well as optical isomers,
e.g. mixtures of enantiomers as well as individual enantiomers and
diastereomers, which arise as a consequence of structural asymmetry
in selected compounds of the present series.
[0138] The compounds of Formula I may also be solvated, especially
hydrated. Hydration may occur during manufacturing of the compounds
or compositions comprising the compounds, or the hydration may
occur over time due to the hygroscopic nature of the compounds.
[0139] Certain compounds within the scope of Formula I are
derivatives referred to as prodrugs. The expression "prodrug"
denotes a derivative of a known direct acting drug, which
derivative has enhanced delivery characteristics and therapeutic
value as compared to the drug, and is transformed into the active
drug by an enzymatic or chemical process; see Notari, R. E.,
"Theory and Practice of Prodrug Kinetics", Methods in Enzymology,
112:309-323 (1985); Bodor, N., "Novel Approaches in Prodrug
Design", Drugs of the Future, 6(3):165-182 (1981); and Bundgaard,
H., "Design of Prodrugs: Bioreversible-Derivatives for Various
Functional Groups and Chemical Entities", in Design of Prodrugs (H.
Bundgaard, ed.), Elsevier, New York (1985). Useful prodrugs are
those where R.sup.a, R.sup.b and/or R.sup.c are --CO.sub.2R.sup.w,
where R.sup.w is defined above. See, U.S. Pat. No. 5,466,811 and
Saulnier et al., Bioorg. Med. Chem. Lett. 4:1985-1990 (1994).
[0140] The term "alkyl" as employed herein by itself or as part of
another group refers to both straight and branched chain radicals
of up to 12 carbons, such as methyl, ethyl, propyl, isopropyl,
butyl, t-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl,
4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl,
undecyl, dodecyl.
[0141] The term "alkenyl" is used herein to mean a straight or
branched chain radical of 2-20 carbon atoms, unless the chain
length is limited thereto, including, but not limited to, ethenyl,
1-propenyl, 2-propenyl, 2-methyl-l-propenyl, 1-butenyl, 2-butenyl,
and the like. Preferably, the alkenyl chain is 2 to 10 carbon atoms
in length, more preferably, 2 to 8 carbon atoms in length most
preferably from 2 to 4 carbon atoms in length.
[0142] The term "alkynyl" is used herein to mean a straight or
branched chain radical of 2-20 carbon atoms, unless the chain
length is limited thereto, wherein there is at least one triple
bond between two of the carbon atoms in the chain, including, but
not limited to, acetylene, 1-propylene, 2-propylene, and the like.
Preferably, the alkynyl chain is 2 to 10 carbon atoms in length,
more preferably, 2 to 8 carbon atoms in length, most preferably
from 2 to 4 carbon atoms in length.
[0143] In all instances herein where there is an alkenyl or alkynyl
moiety as a substituent group, the unsaturated linkage, i.e., the
vinylene or acetylene linkage is preferably not directly attached
to a nitrogen, oxygen or sulfur moiety.
[0144] The term "alkoxy" is used herein to mean a straight or
branched chain radical of 1 to 20 carbon atoms, unless the chain
length is limited thereto, bonded to an oxygen atom, including, but
not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, and the
like. Preferably the alkoxy chain is 1 to 10 carbon atoms in
length, more preferably 1 to 8 carbon atoms in length.
[0145] The term "aryl" as employed herein by itself or as part of
another group refers to monocyclic or bicyclic aromatic groups
containing from 6 to 12 carbons in the ring portion, preferably
6-10 carbons in the ring portion, such as phenyl, naphthyl or
tetrahydronaphthyl.
[0146] The term "heteroaryl" as employed herein refers to groups
having 5 to 14 ring atoms; 6, 10 or 14 .pi. electrons shared in
acyclic array; and containing carbon atoms and 1, 2 or 3 oxygen,
nitrogen or sulfur heteroatoms (where examples of heteroaryl groups
are: thienyl, benzo[b]thienyl, naphtho[2,3-b]thienyl, thianthrenyl,
furyl, pyranyl, isobenzofliranyl, benzoxazolyl, chromenyl,
xanthenyl, phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl,
pyrazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl,
indolizinyl, isoindolyl, 3H-indolyl, indolyl, indazolyl, purinyl,
4H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl,
naphthyridinyl, quinazolinyl, cinnolinyl, pteridinyl,
4aH-carbazolyl, carbazolyl, .beta.-carbolinyl, phenanthridinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, isothiazolyl,
phenothiazinyl, isoxazolyl, furazanyl and phenoxazinyl groups).
[0147] The term "aralkyl" or "arylalkyl" as employed herein by
itself or as part of another group refers to C.sub.1-6alkyl groups
as discussed above having an aryl substituent, such as benzyl,
phenylethyl or 2-naphthylmethyl.
[0148] The term "cycloalkyl" as employed herein by itself or as
part of another group refers to cycloalkyl groups containing 3 to 9
carbon atoms. Typical examples are cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and
cyclononyl.
[0149] The terms "alkoxy" refers to any of the above alkyl groups
linked to an oxygen atom.
[0150] The term "halogen" or "halo" as employed herein by itself or
as part of another group refers to chlorine, bromine, fluorine or
iodine with chlorine being preferred.
[0151] The term "monoalkylamine" as employed herein by itself or as
part of another group refers to an amino group which is substituted
with one alkyl group having from 1 to 6 carbon atoms.
[0152] The term "dialkylamine" as employed herein by itself or as
part of another group refers to an amino group which is substituted
with two alkyl groups, each having from 1 to 6 carbon atoms
[0153] The term "hydroxyalkyl" as employed herein refers to any of
the above alkyl groups substituted by one or more hydroxyl
moieties.
[0154] The term "carboxyalkyl" as employed herein refers to any of
the above alkyl groups substituted by one or more carboxylic acid
moieties.
[0155] The term "heterocyclic" is used herein to mean a saturated
or wholly or partially unsaturated 3-7 membered monocyclic, or 7-10
membered bicyclic ring system, which consists of carbon atoms and
from one to four heteroatoms independently selected from the group
consisting of O, N, and S, wherein the nitrogen and sulfur
heteroatorns can be optionally oxidized, the nitrogen can be
optionally quaternized, and including any bicyclic group in which
any of the above-defined heterocyclic rings is fused to a benzene
ring, and wherein the heterocyclic ring can be substituted on
carbon or on a nitrogen atom if the resulting compound is stable.
Especially useful are rings containing one oxygen or sulfur, one to
three nitrogen atoms, or one oxygen or sulfur combined with one or
two nitrogen atoms. Examples of such heterocyclic groups include
piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl,
2-oxopyrrolodinyl, 2-oxoazepinyl, azepinyl, pyrrolyl,
4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl,
imidazolinyl, imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl,
pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl,
morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl,
isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl,
tetrahydroisoquinolinyl, benzimidazolyl, thiadiazoyl, benzopyranyl,
benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl,
tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl,
thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and
oxadiazolyl. Morpholino is the same as morpholinyl.
[0156] The term "heteroatom" is used herein to mean an oxygen atom
("O"), a sulfur atom ("S") or a nitrogen atom ("N"). It will be
recognized that when the heteroatom is nitrogen, it may form an
NR.sup.yR.sup.z moiety, wherein R.sup.y and R.sup.z are,
independently from one another, hydrogen or C.sub.1 to C.sub.8
alkyl, or together with the nitrogen to which they are bound, form
a saturated or unsaturated 5-, 6-, or 7-membered ring.
[0157] The term "heteroaryl" includes 5 or 6 membered aromatic
heterocyclic rings containing one or more heteroatoms selected from
nitrogen, sulphur and oxygen atoms, and fused bicyclic ring systems
containing one or more nitrogen, sulfur, and oxygen atoms. Examples
of such groups include oxadiazole, thiazole thiadiazole, triazole,
tetrazole, benzimidazole, pyridine, furan and thiophene.
[0158] A C.sub.3-7 cycloalkenyl group includes rings containing at
least one double bond incorporated in the ring.
[0159] A C.sub.3-7 heterocycloalkyl group includes rings containing
one or more heteroatoms selected from nitrogen, sulphur and oxygen
atoms, for example, a tetrahydropyran-4-yl group.
[0160] A C.sub.3-7 heterocycloalkenyl group includes rings
containing one or more heteroatoms selected from nitrogen, sulphur
and oxygen atoms, together with at least on double bond
incorporated in the ring.
[0161] Another aspect of the present invention is a process for
preparing an aminoguanidine compound ofFormulaI, comprising
reacting an aminoguanidine of the formula: 19
[0162] wherein R.sup.11, R.sup.a, R.sup.b and R.sup.c are defined
as above, with a carbonyl-containing compound of the formula:
20
[0163] wherein R.sup.1-R.sup.6, Y, n, m, R.sup.7, R.sup.8, R.sup.9
and R.sup.10 are defined as above to form an amidinohydrazone, and
thereafter selectively reducing the hydrazone carbon to nitrogen
double bond of the amidinohydrazone.
[0164] The aminoguanidine is typically provided as a salt,
preferably the nitrate salt. The first step proceeds at ambient
temperature using alcohol as a solvent. An acid, such as 4 N HCl in
dioxane is added to the reaction mixture.
[0165] Another aspect of the present invention is a process for
preparing a hydroxyguanidine compound of Formula I, comprising
reacting an alkoxyamine compound of the formula: 21
[0166] wherein R.sup.1-R.sup.6, Y, n, m, R.sup.7, R.sup.8, R.sup.9
and R.sup.10 are defined as above with a guanidinylating reagent.
Preferred guanidinylating reagents include: aminoiminosulfonic
acid, optionally substituted 1 H-pyrazole- 1-carboxamidines, or
N,N'-bis(tert-butoxycarbon- yl) S-methyl isothiourea.
[0167] Schemes 1a, 1b and 1c exemplify the synthetic steps to
produce compounds of the present invention. 22
[0168] Halogenated phenols 1 may be monoprotected (P.sup.a is a
protecting group) with a variety of protecting groups known in the
art, such as esters and benzyl ethers (Greene, T. W. and Wuts, P.
G. M., Protective Groups in Organic Synthesis, 2nd edition, John
Wiley and Sons, Inc. New York (1991)). Deprotection of the hydroxy
groups is routinely accomplished using the reaction conditions well
known in the art. For example, deprotection of benzyl ethers may be
effected through catalytic hydrogenation using palladium on carbon
as a catalyst in solvents such as ethanol or tetrahydrofuran.
Deprotection of an acetate is accomplished by basic hydrolysis,
most preferably with sodium hydroxide in aqueous
tetrahydrofuran.
[0169] Additional compounds ofthe invention are formed by employing
phenols that are halogenated ortho- orpara- to the hydroxy groups
in place of the meta-halogenated phenols 1.
[0170] Halogenated phenols 1 are carboxylated to form phenolic
carboxylic acids 2, which are then reacted with suitable amines to
form phenolic amides 3. Phenolic amides 3 are coupled to 4 (for
L=OH) using a Mitsunobu coupling procedure (Mitsunobu, O.,
Synthesis 1 (1981)), where P.sup.b of 4 may be a suitable alcohol
protecting group. Alternatively, suitable diols (P.sup.b=H) may be
used in the Mitsunobu reaction. Preferred coupling conditions
include using a trialkylphosphine or triarylphosphine, such as
triphenylphosphine or tri-n-butylphosphine, in a suitable solvent,
such as tetrahydrofuran or dichloromethane, and an azodicarbonyl
reagent, such as diethyl azodicarboxylate or
1,1'-(azodicarbonyl)dipiperidine. Typical P.sup.b (where P.sup.b is
an alcohol protecting group) is well known in the art, such as
esters and benzyl ethers (Greene, T. W. and Wuts, P. G. M., supra).
Alternatively, where L is a reactive leaving group such as halide
or sulfonate, phenol 3 may be treated with a base, such as sodium
hydride, in a solvent, such as N,N-dimethylformamide, and then
treated with 4. Removal of P.sup.b is routinely accomplished using
the reaction conditions well known in the art. For example,
deprotection of benzyl ethers may be effected through catalytic
hydrogenation using palladium on carbon as a catalyst in solvents
such as ethanol or tetrahydrofuran. Deprotection of an acetate is
accomplished by basic hydrolysis, most preferably with sodium
hydroxide in aqueous tetrahydrofuran.
[0171] Alcohol 5 is converted to 6 employing a Mitsunobu reaction
with an N-hydroxycyclic imide derivative such as
N-hydroxyphthalimide. Unveiling of the phthalimide protecting group
is accomplished using standard conditions well known in the art
(Greene, T. W. and Wuts, P. G. M., supra), for example, sodium
borohydride in a mixture of an appropriate alcohol (e.g. ethanol or
2-propanol)/water followed by acidification. Alternatively, removal
of the protecting group may be accomplished using hydrazine or
methylamine.
[0172] Guanidinylation of the resulting alkoxyamine to 7 is
achieved using standard reagents such as aminoiminosulfonic acid
(Miller, A. E. and Bischoff, J. J. Synthesis 777 (1986)), or
1H-pyrazole-1-carboxamidine hydrochloride (Bernatowicz, M. S. et.
al. J. Org. Chem 57(8):2497 (1992)), or with substituted
guanidinylating reagents such as
N,N'-bis(tert-butoxycarbonyl)-S-methylisothiourea (Bergeron, R. J.
and McManis, J. S. J. Org. Chem. 52:1700 (1987)) or N-R.sup.a,
N-R.sup.b, N'-R.sup.c-1H-pyrazole-1-carboxamidine, where R.sup.a,
R.sup.b and R.sup.c are defined as above for Formula I Useful
1H-pyrazole-1-carboxami- dines include
N,N'-bis(tert-butoxycarbonyl)-1H-pyrazole-1-carboxamidine and
N,N'-bis(benzyloxycarbonyl)- 1H-pyrazole- 1-carboxamidine (all
ofwhich can be prepared according to Bernatowicz, M. S. et. al.,
Tetrahedron Letters 34:3389 (1993)).
[0173] Alkoxyguanidines (where R.sup.11 is H) may be optionally
alkylated using such reagents as alkyl bromides, and bases such as
sodium bicarbonate, in a solvent such as N,N-dimethylformamide to
form compounds where R.sup.11 is alkyl.
[0174] Scheme 1b describes an alternative synthesis for forming
compounds where X is NH. Phenolic amide 3 may be converted to 9 by
the Mitsunobu reaction using 8 wherein L=OH and the P.sup.c's are
alkyl groups or are combined to form a cycloalkyl or cycloalkenyl
group. Alternatively, where L of 8 is a reactive leaving group such
as halide or sulfonate, phenol 3 may be treated with a base, such
as sodium hydride in a solvent such as N,N-dimethylformamide, and
then treated with 8. Protecting groups, P.sup.c, may then be
removed to afford 9 using standard conditions well known in the
art, for example, p-toluenesulfonic acid in acetone (Greene, T. W.
and Wuts, P. G. M., supra).
[0175] Compound 9 is then converted to amidinohydrazone 11 using
standard conditions, for example, treatment with an aminoguanidine,
such as aminoguanidine or 2-hydrazinoimidazoline, optionally in the
presence of an acid such as nitric acid, hydrogen chloride, or
hydrogen bromide, in an appropriate solvent, for example, ethanol
or methanol, which, in addition, may contain other solvents such as
dichloromethane or tetrahydrofuran. Conversion of 11 to 12 is
accomplished under reducing conditions well known in the art, for
example, lithium borohydride in an appropriate solvent such as
tetrahydrofuran or methanol at various temperatures up to reflux.
As an alternative method, catalytic hydrogenation with palladium on
carbon catalyst can be employed.
[0176] When R.sup.a, R.sup.b and/or R.sup.c are a protecting group,
for example tert-butyloxycarbonyl (Boc), these protecting groups
can be optionally removed by treatment with acid, usually
trifluoroacetic acid in a suitable solvent such as dichloromethane
or water, or by HCl gas dissolved in a suitable solvent. such as
1,4-dioxane.
[0177] Scheme 1c describes an alternative synthesis that can be
used to generate libraries of compounds 7 in parallel. Carboxylic
acid 2 may be protected with a protecting group (P.sup.c), such as
a benzyl ester, and the P.sup.a group (described above) removed
with a reagent, such as tetrabutylammonium fluoride, both well
known in the art (Green, T. W., and Wuts, P. G. M., supra) giving
phenol 33. Phenol 33, where L is a reactive leaving group such as
halide or sulfonate, can then be treated with a base, such as
cesium carbonate, in a solvent, such as acetonitrile, and reacted
with 4. The P.sup.b group may then be removed as above to form
alcohol 34, which can be converted to 35 employing a Mitsunobu
reaction with an N-hydroxycyclic imide derivative such as
N-hydroxyphthalimide. The phthalimide protecting group can be
removed and the resulting alkoxyamine can be guanidinylated as
above, and the P.sup.c group (e.g., benzyl ester) may then be
removed to afford 36 using standard conditions well known in the
art, for example, aqueous sodium hydroxide in ethanol (Greene, T.
W., and Wuts, P. G. M., supra). Carboxylic acid 36 can then be
coupled to a variety of different amines and purified in a parallel
format giving a library of compounds 7.
[0178] As an alternative scheme to produce the
O-phthalamide-containing intermediates 6, the respective phenolic
amides 3 may be reacted under basic conditions with reagent 23
which contains a leaving group L' (Scheme 2). This scheme is
limited to producing compounds where R.sup.10 is hydrogen. Reagent
23 is produced by reacting a compound (22) having two leaving
groups, L and L', under basic conditions with N-hydroxyphthalimide
(Khadilkar and Samant, Indian J Chem. Sec. B 1137 (1993)).
[0179] Compounds wherein R.sup.7 and R.sup.10 (III) or R.sup.8 and
R.sup.10 (IV) together form a methylene linkage can be synthesized
by substituting a cyclic ketone having a reactive group L that is
attached directly or indirectly to the carbocyclic ring. Examples
of suitable reagents include 2-hydroxycyclopentanone,
3-hydroxycyclopentanone, 2-hydroxycyclohexanone and 3
-hydroxycyclohexanone. 23 24
[0180] Compounds VI wherein R.sup.11 and R.sup.b are taken together
with the nitrogens to which they are attached to form a ring
structure are prepared by substituting a heterocyclic amine X
(below) for the aminoguanidine in the above Schemes. 25
[0181] Compounds V wherein R.sup.a and R.sup.c are taken together
with the nitrogen atoms to which they are attached to form an
imidazoline moiety are prepared by substituting a
2-hydrazinoimidazo line XI (above) for the aminoguanidines in the
above Schemes.
[0182] For medicinal use, the pharmaceutically acceptable acid
addition salts, those salts in which the anion does not contribute
significantly to toxicity or pharmacological activity of the
organic cation, are preferred. The acid addition salts are obtained
either by reaction of an organic base of Formula I with an organic
or inorganic acid, preferably by contact in solution, or by any of
the standard methods detailed in the literature available to any
practitioner skilled in the art. Examples of useful organic acids
are carboxylic acids such as maleic acid, acetic acid, tartaric
acid, propionic acid, fumaric acid, isethionic acid, succinic acid,
cyclamic acid, pivalic acid and the like; useful inorganic acids
are hydrohalide acids such as HCl, HBr, HI; sulfuric acid;
phosphoric acid and the like. Preferred acids for forming acid
addition salts include HCl and acetic acid.
[0183] The compounds of the present invention represent a novel
class of potent inhibitors of metallo, acid, thiol and serine
proteases. Examples of the serine proteases inhibited by compounds
within the scope of the invention include leukocyte neutrophil
elastase, a proteolytic enzyme implicated in the pathogenesis of
emphysema; chymotrypsin and trypsin, digestive enzymes; pancreatic
elastase, and cathepsin G, a chymotrypsin-like protease also
associated with leukocytes; thrombin and factor Xa, proteolytic
enzymes in the blood coagulation pathway. Inhibition ofthermolysin,
a metalloprotease, and pepsin, an acid protease, are also
contemplated uses of compounds of the present invention. The
compounds of the present invention are preferably employed to
inhibit trypsin-like proteases.
[0184] An end use application of the compounds that inhibit
chymotrypsin and trypsin is in the treatment of pancreatitis. For
their end-use application, the It !; potency and other biochemical
parameters of the enzyme-inhibiting characteristics of the
compounds of the present invention is readily ascertained by
standard biochemical techniques well known in the art. Actual dose
ranges for their specific end-use application will, of course,
depend upon the nature and severity of the disease state of the
patient or animal to be treated, as determined by the attending
diagnostician. It is expected that a useful dose range will be
about 0.01 to 10 mg per kg per day for an effective therapeutic
effect.
[0185] Compounds ofthe present invention that are distinguished by
their ability to inhibit either factor Xa or thrombin may be
employed for a number of therapeutic purposes. As factor Xa or
thrombin inhibitors, compounds of the present invention inhibit
thrombin production. Therefore, these compounds are useful for the
treatment or prophylaxis of states characterized by abnormal venous
or arterial thrombosis involving either thrombin production or
action. These states include, but are not limited to, deep vein
thrombosis; disseminated intravascular coagulopathy which occurs
during septic shock, viral infections and cancer; myocardial
infarction; stroke; coronary artery bypass; fibrin formation in the
eye; hip replacement; and thrombus formation resulting from either
thrombolytic therapy or percutaneous transluminal coronary
angioplasty (PCTA).
[0186] Other uses include the use of said thrombin inhibitors as
anticoagulants either embedded in or physically linked to materials
used in the manufacture of devices used in blood collection, blood
circulation, and blood storage, such as catheters, blood dialysis
machines, blood collection syringes and tubes, blood lines and
stents. The compounds of the present invention may also be used as
an anticoagulant in extracorporeal blood circuits.
[0187] Metal stents have been shown to reduce restenosis, but are
thrombogenic. A strategy for reducing the thrombogenicity of stents
is to coat, embed, adsord or covalently attach a
thrombin-inhibiting agent to the stent surface. The compounds of
the present invention can be employed for this purpose. Compounds
of the invention can be attached to, or embedded within soluble
and/or biodegradeable polymers as and thereafter coated onto stent
materials. Such polymers can include polyvinylpyrrolidone,
polyhydroxy-propyl-methacrylamide-phenol,
polyhydroxyethyl-aspartamide-phenol, or
polyethyleneoxide-polylysine substituted with palmitoyl residues,
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. See European Application 761 251, European
Application 604,022, Canadian Patent 2,164,684 and PCT Published
Applications WO 96/11668, WO 96/32143 and WO 96/38136.
[0188] By virtue of the effects of both factor Xa and thrombin on a
host of cell types, such as smooth muscle cells, endothelial cells
and neutrophils, the compounds of the present invention find
additional use in the treatment or prophylaxis of adult respiratory
distress syndrome; inflammatory responses; wound healing;
reperfusion damage; atherosclerosis; and restenosis following an
injury such as balloon angioplasty, atherectomy, and arterial stent
placement. The compounds of the present invention may be useful in
treating neoplasia and metastasis as well as neurodegenerative
diseases, such as Alzheimer's disease and Parkinson's disease.
[0189] When employed as thrombin or factor Xa inhibitors, the
compounds of the present invention may be administered in an
effective amount within the dosage range of about 0.1 to about 500
mg/kg, preferably between 0.1 to 10 mg/kg body weight, on a regimen
in single or 2-4 divided daily doses.
[0190] When employed as inhibitors of thrombin, the compounds of
the present invention may be used in combination with thrombolytic
agents such as tissue plasminogen activator, streptokinase, and
urokinase. Additionally, the compounds of the present invention may
be used in combination with other antithrombotic or anticoagulant
drugs such as, but not limited to, fibrinogen antagonists and
thromboxane receptor antagonists.
[0191] Human leucocyte elastase is released by polymorphonuclear
leukocytes at sites of inflammation and thus is a contributing
cause for a number of disease states. Compounds of the present
invention are expected to have an anti- inflammatory effect useful
in the treatment of gout, rheumatoid arthritis and other
inflammatory diseases, and in the treatment of emphysema. The
leucocyte elastase inhibitory properties of compounds of the
present invention are determined by the method described below.
Cathepsin G has also been implicated in the disease states of
arthritis, gout and emphysema, and in addition, glomerulonephritis
and lung infestations caused by infections in the lung. In their
end-use application the enzyme inhibitory properties of the
compounds of Formula I is readily ascertained by standard
biochemical techniques that are well-known in the art.
[0192] The Cathepsin G inhibitory properties of compounds within
the scope of the present invention are determined by the following
method. A preparation of partially purified human Cathepsin G is
obtained by the procedure of Baugh et al., Biochemistry 15: 836
(1979). Leukocyte granules are a major source for the preparation
of leukocyte elastase and cathepsin G (chymotrypsin-like activity).
Leukocytes are lysed and granules are isolated. The leukocyte
granules are extracted with 0.20 M sodium acetate, pH 4.0, and
extracts are dialyzed against 0.05 M Tris buffer, pH 8.0 containing
0.05 M NaCl overnight at 4.degree. C. A protein fraction
precipitates during dialysis and is isolated by centrifugation.
This fraction contains most of the chymotrypsin-like activity of
leukocyte granules. Specific substrates are prepared for each
enzyme, namely N-Suc-Ala-Ala-Pro-Val-p-nitroanilide and
Suc-Ala-Ala-Pro-Phe-p-nit- roanilide. The latter is not hydrolyzed
by leukocyte elastase. Enzyme preparations are assayed in 2.00 mL
of 0.10 M Hepes buffer, pH 7.5, containing 0.50 M NaCl, 10%
dimethylsulfoxide and 0.0020 M Suc-Ala-Ala-Pro-Phe-p-nitroanilide
as a substrate. Hydrolysis of the p-nitroanilide substrate is
monitored at 405 nm and at 25.degree. C.
[0193] Useful dose range for the application of compounds of the
present invention as neutrophil elastase inhibitors and as
Cathepsin G inhibitors depend upon the nature and severity of the
disease state, as determined by the attending diagnostician, with a
range of 0.01 to 10 mg/kg body weight, per day, being useful for
the aforementioned disease states.
[0194] Compounds ofthe present invention that inhibit urokinase or
plasminogen activator are potentially useful in treating excessive
cell growth disease state. As such compounds of the present
invention may also be useful in the treatment of benign prostatic
hypertrophy and prostatic carcinoma, the treatment of psoriasis,
and as abortifacients. For their end-use application, the potency
and other biochemical parameters of the enzyme inhibiting
characteristics of compounds of the present invention are readily
ascertained by standard biochemical techniques well known in the
art. Actual dose ranges for this application will depend upon the
nature and severity of the disease state of the patient or animal
to be treated as determined by the attending diagnostician. It is
to be expected that a general dose range will be about 0.01 to 10
mg per kg per day for an effective therapeutic effect.
[0195] Additional uses for compounds of the present invention
include analysis of commercial reagent enzymes for active site
concentration. For example, chymotrypsin is supplied as a standard
reagent for use in clinical quantitation of chymotrypsin activity
in pancreatic juices and feces. Such assays are diagnostic for
gastrointestinal and pancreatic disorders. Pancreatic elastase is
also supplied commercially as a reagent for quantitation ofa,
-antitrypsin in plasma. Plasma .alpha..sub.1-antitrypsin increases
in concentration during the course of several inflammatory
diseases, and .alpha..sub.1-antitrypsin deficiencies are associated
with increased incidence of lung disease. Compounds of the present
invention can be used to enhance the accuracy and reproducibility
of these assays by titrametric standardization of the commercial
elastase supplied as a reagent. See, U.S. Pat. No. 4,499,082.
[0196] Protease activity in certain protein extracts during
purification of particular proteins is a recurring problem which
can complicate and compromise the results of protein isolation
procedures. Certain proteases present in such extracts can be
inhibited during purification steps by compounds of the present
invention, which bind tightly to various proteolytic enzymes.
[0197] The pharmaceutical compositions ofthe invention can be
administered to any animal that can experience the beneficial
effects of the compounds of the invention. Foremost among such
animals are humans, although the invention is not intended to be so
limited.
[0198] The pharmaceutical compositions of the present invention can
be administered by any means that achieve their intended purpose.
For example, administration can be by parenteral, subcutaneous,
intravenous, intramuscular, intraperitoneal, transdermal, buccal,
or ocular routes. Alternatively. or concurrently, administration
can be by the oral route. The dosage administered will be dependent
upon the age, health, and weight of the recipient, kind of
concurrent treatment, if any, frequency of treatment, and the
nature of the effect desired.
[0199] In addition to the pharmacologically active compounds, the
new pharmaceutical preparations can contain suitable
pharmaceutically acceptable carriers comprising excipients and
auxiliaries that facilitate processing of the active compounds into
preparations that can be used pharmaceutically.
[0200] The pharmaceutical preparations of the present invention are
manufactured in a manner that is, itself, known, for example, by
means of conventional mixing, granulating, dragee-making,
dissolving, or lyophilizing processes. Thus, pharmaceutical
preparations for oral use can be obtained by combining the active
compounds with solid excipients, optionally grinding the resulting
mixture and processing the mixture of granules, after adding
suitable auxiliaries, if desired or necessary, to obtain tablets or
dragee cores.
[0201] Suitable excipients are, in particular, fillers such as
saccharides, for example, lactose or sucrose, mannitol or sorbitol,
cellulose preparations and/or calcium phosphates, for example,
tricalcium phosphate or calcium hydrogen phosphate, as well as
binders, such as, starch paste, using, for example, maize starch,
wheat starch, rice starch, potato starch, gelatin, tragacanth,
methyl cellulose, hydroxypropylmethylcellulose, sodium
carboxymethylcellulose, and/or polyvinyl pyrrolidone. If desired,
disintegrating agents can be added, such as, the above-mentioned
starches and also carboxymethyl-starch, cross-linked polyvinyl
pyrrolidone, agar, or alginic acid or a salt thereof, such as,
sodium alginate. Auxiliaries are, above all, flow-regulating agents
and lubricants, for exarnple, silica, talc, stearic acid or salts
thereof, such as, magnesium stearate or calcium stearate, and/or
polyethylene glycol. Dragee cores are provided with suitable
coatings that, if desired, are resistant to gastric juices. For
this purpose, concentrated saccharide solutions can be used, which
may optionally contain gum arabic, talc, polyvinyl pyrrolidone,
polyethylene glycol, and/or titanium dioxide, lacquer solutions and
suitable organic solvents or solvent mixtures. In order to produce
coatings resistant to gastric juices, solutions of suitable
cellulose preparations, such as, acetylcellulose phthalate or
hydroxypropylmethyl-cellulose phthalate, are used. Dye stuffs or
pigments can be added to the tablets or dragee coatings, for
example, for identification or in order to characterize
combinations of active compound doses.
[0202] Other pharmaceutical preparations which can be used orally
include push-fit capsules made of gelatin, as well as soft, sealed
capsules made of gelatin and a plasticizer, such as, glycerol or
sorbitol. The push-fit capsules can contain the active compounds in
the form of granules that may be mixed with fillers such as
lactose, binders such as starches, and/or lubricants such as talc
or magnesium stearate and, optionally, stabilizers. In soft
capsules, the active compounds are preferably dissolved or
suspended in suitable liquids, such as, fatty oils or liquid
paraffin. In addition, stabilizers may be added.
[0203] Suitable formulations for parenteral administration include
aqueous solutions of the active compounds in water-soluble form,
for example, water-soluble salts, alkaline solutions and
cyclodextrin inclusion complexes. Especially preferred salts are
hydrochloride and acetate salts. One or more modified or unmodified
cyclodextrins can be employed to stabilize and increase the water
solubility of compounds of the present invention. Useful
cyclodextrins for this purpose are disclosed in U.S. Pat. Nos.
4,727,064, 4,764,604, and 5,024,998.
[0204] In addition, suspensions of the active compounds as
appropriate oily injection suspensions can be administered.
Suitable lipophilic solvents or vehicles include fatty oils, for
example, sesame oil, or synthetic fatty acid esters, for example,
ethyl oleate or triglycerides or polyethylene glycol-400 (the
compounds are soluble in PEG-400). Aqueous injection suspensions
can contain substances that increase the viscosity of the
suspension, for example, sodium carboxymethyl cellulose, sorbitol,
and/or dextran. Optionally, the suspension may also contain
stabilizers.
[0205] In another aspect, the present invention includes
compositions which are useful for in vivo imaging of thrombi in a
mammal, comprising a compound of the present invention which is
capable of being detected outside the body. Preferred are
compositions comprising a compound of the present invention and a
detectable label, such as a radioactive or paramagnetic atom.
[0206] In another aspect, the present invention provides diagnostic
compositions which are use for in vivo imaging of thrombi in a
mammal, comprising a pharmaceutically acceptable carrier and a
diagnostically effective amount of a compound or composition of the
present invention.
[0207] In another aspect, the present invention includes methods
which are useful for in vivo imaging or thrombi in a mammal.
[0208] According to a preferred aspect, useful compounds are those
wherein the R.sup.1 substituent is substituted with a detectable
label, such as a radioactive iodine atom, such as I-125,I-131 or
I-123. In this aspect, R.sup.1 is preferably phenyl, having a para
I-123, para I-125 or para I-131 substitution.
[0209] The detectable label can also be a radioactive or
paramagnetic chelate in which a suitable ligand (L) is attached to
an R.sup.1 substituent, either directly or via a divalent linking
group A". Alternatively, the group -A"-L substitutes for the groups
-Z-R.sup.1 in Formula I By suitable ligand is meant an organic
moiety that is capable of chelating a radioactive or paramagnetic
metal ion.
[0210] In these compounds, the divalent linking group A" includes
groups that are capable of covalently bonding with a free amino
group and the chelating means. For example, A" may be
--C(.dbd.S)--, --C(.dbd.O)--,
--C(.dbd.NH)--(CH.sub.2).sub.6--C(.dbd.NH)--,
--C(.dbd.O)--(CH.sub.2).sub- .6--C(.dbd.O)--, 26
[0211] and the like.
[0212] Also, in the compounds represented by Formula I, the
chelating ligand, L, includes groups capable of covalently bonding
to or noncovalently binding to either a radioactive or paramagnetic
atom. The chelating means including those which are customarily
used for complexing radioactive or paramagnetic atoms. These
include chelating means containing 3 to 12, preferably 3 to 8,
methylene phosphonic acid groups, methylene carbohydroxamic acid
groups, carboxyethylidene groups, or especially carboxymethylene
groups, which are bonded to a nitrogen atom. If only one or two of
the acid groups are bonded to a nitrogen atom, then that nitrogen
is bonded to another nitrogen atom having such groups by an
optionally substituted ethylene groups or by up to four separated
ethylene units separated by a nitrogen or oxygen or sulfuir atom.
Preferred as acompleting means is
diethylenetrimine-N,N,N',N",N"-pentaace- tic acid (DTPA). DTPA is
well known in the art as a chelating means for the radioactive atom
indium-111 (In-111), technetium-99m (Tc-99m), and the paramagnetic
atom gadoliniuni (Gd). Khaw, et al., Science 209:295 (1980); Paik
C. H. et al., U.S. Pat. No. 4,652,440 (1987); Gries, H. et al.,
U.S. Pat. No. 4,957,939 (1990). An preferred chelating ligand, L,
is i-(p-aminobenzyl)-diethylenetriaminepentaacetic acid. Also
included as chelating means are compounds which contain sulfhdryl
or amine moieties, the total of which in any combination is at
least four. These sulfhydryl or amine moieties are separated from
each other by at least two atoms which can be either carbon,
nitrogen, oxygen, or sulfur. Especially preferred for chelating
means, L, is metallothionein which is well known in the art as a
chelating means for Tc-99m.
[0213] Compounds of Formula I can be labeled with radioactive
halogen atom by using an appropriate exchange reaction. Exchange of
hot iodine for cold iodine is well known in the art. Alternatively,
a radio iodine labeled compound can be prepared from the
corresponding bromo compound via a tributylstannyl intermediate.
See, U.S. Pat. No. 5,122,361, herein incorporated by reference.
[0214] The present invention also includes compositions which are
useful for in vivo imaging of thrombi in a mammal, wherein the
compositions are comprised of a compound of Formula I complexed
with a radioactive atom.
[0215] For the compounds of Formula I, suitable radioactive atoms
include Co-57, Cu-67, Ga-67, Ga-68, Ru-97, Tc-99m, In-111, In-113m,
Hg- 197, Au-198. and Pb-203. Some radioactive atoms have superior
properties for use in radiochemical imaging techniques. In
particular, technetium-99m (Tc-99m) is an ideal radioactive atom
for imaging because of its nuclear properties. It is a gamma
emitter and has a single photon energy of 140 ke V, a half-life of
about 6 hours, and it is readily available from a Mo-99/Tc-99
generator. Rhenium-186 and -188 also have gamma emission which
allows it to be imaged. Preferred compositions contain the
radioactive atom, Tc-99m.
[0216] Compositions of the present invention are conveniently
prepared by completing a compound of Formula I with radioisotopes
which are suitable for detection externally. The gamma emitters,
indium-111m and technetium-99m, are preferred as radioactive atoms
because they are detectable with a gamma camera and have favorable
half-lives in vivo.
[0217] The compounds of Formula I can be labeled by any of the many
techniques known in the art to provide a composition of the present
invention. For example, these compounds can be labeled through a
chelating agent such as diethylene-triaminepentaacetic acid (DTPA)
or metallothionein, both of which can be covalently attached to the
compound of Formula I via a bond to the R.sup.1 or R.sup.2 group
that will be outside the binding pocket of thrombin.
[0218] In general, the compositions of the present invention
containing technetium-99m are prepared by forming an aqueous
mixture oftechnetium-99m and a reducing agent and a water-soluble
ligand, and then contacting the mixture with a compound of the
present invention represented by Formula L For example, the imaging
compounds of this invention are made by reacting technetium-99m (in
an oxidized state) with the compounds of the present invention
having a chelating means in the presence of a reducing agent to
form a stable complex between technetium-99m in a reduced state (IV
or V valence state).
[0219] One embodiment of the composition of the present invention
is prepared by labeling a compound of Formula I having a DTPA
chelating means with technetium-99m. This may be accomplished by
combining a predetermined amount (as 5 .mu.g to 0.5 mg) of compound
of the present invention with an aqueous solution containing
citrate buffer and stannous reducing agent, then adding freshly
eluted sodium pertechnetate containing a predetermined level of
radioactivity (as 15 mCi). After allowing an incubation of the
mixture at room temperature, the reaction mixture is loaded into a
shielded syringe through a sterile filter (0.2-0.22 micron), then
is dispensed into 0.9% saline for injection, if desired.
[0220] Another embodiment of the compositions of the present
invention is prepared by labeling acompound ofFormulaIhaving
ametallothionein chelating means with technetium-99m. This may be
accomplished by combining aqueous sodium pertechnetate-99m with
aqueous stannous glucoheptonate to form a soluble complex of
technetium-99m (in reduced state) with two glucoheptonate
molecules, then combining this solution with a compound of the
Formula I having a metallothionein attached thereto. After
incubating the mixture for a period of time and under conditions
which allow for an exchange of the technetium-99m from the
glucoheptonate complex to the metallothionein of the compound of
Formula I, the technetium-labeled composition of the present
invention is formed.
[0221] The source of technetium-99m should preferably be water
soluble. Preferred sources are alkali and alkaline earth metal
pertechnetate (TcO.sub.4.sup.13 ). Technetium-99m is most
preferably obtained in the form of fresh sodium pertechnetate from
a sterile technetium-99m generator (as from a conventional
Mo-99/Tc-99m generator). However, any other source of
physiologically acceptable technetium-99m may be used.
[0222] Reducing agents for use in the method are physiologically
acceptable for reducing technetium-99m from its oxidized state to
the IV or V valence state or for reducing rhenium from its oxidized
state. Reducing agents which can be used are stannous chloride,
stannous fluoride, stannous glucoheptonate, stannous tartarate, and
sodium dithionite. The preferred agents are stannous reducing
agents, especially stannous chloride or stannous glucoheptonate.
The amount of reducing agent is that amount necessary to reduce the
technetium-99m to provide for the binding to the chelating means of
a compound of Formula I in this radioisotope's reduced state. For
example, stannous chloride (SnCl.sub.2) is the reducing agent and
can be used in range from 1-1,000 ,.mu.g/mL. Especially preferred
concentrations are about 30-500 .rho.g/mL.
[0223] Citric acid complexes with technetium-99m quickly to form a
stable technetium-99m-citrate complex. Upon contact with a compound
of Formula I, substantially quantitative transfer of technetium-99m
from its citrate complex to the chelating means of the compound of
Formula I is achieved rapidly and under mild conditions. The amount
of citric acid (as sodium citrate) can range from about 0.5 mg/ml
up to the amount maximally soluble in the medium. Preferred amounts
of citric acid range from 15 to 30.mu.g/ml.
[0224] The amount of compound of Formula I having a chelating means
can range from 0.001 to about 3 mg/mL, preferably about 0.017 to
about 0.15 mg/mL. Finally, technetium-99m in the form of
pertechnetate can be used in amounts of preferably about 1-50 mCi.
The amount of mCi per mg of compound of the present invention is
preferably about 30-150.
[0225] The reaction between the compound of Formula I and the metal
ion-transfer ligand complex is preferably carried out in a aqueous
solution at a pH at which the compound of Formula I is stable. By
"stable", it is meant that the compound remains soluble and retains
its inhibitory activity against .alpha.-thrombin. Normally, the pH
for the reaction will be from about 5 to 9, the preferred pH being
above 6-8. The technetium-99m-citrate complex and a compound of
Formula I are incubated, preferably at a temperature from about
20.degree. C. to about 60.degree. C., most preferably from about
20.degree. C. to about 37.degree. C., for a sufficient amount of
time to allow transfer ofthe metal ion from the citrate complex to
the chelating means of the compound of Formula I. Generally, less
than one hour is sufficient to complete the transfer reaction under
these conditions.
[0226] The present invention also includes compositions ofthe
compounds ofthe present invention which are useful for in vivo
imaging of thrombi in a mammal, comprised of a compound represented
by Formula I complexed to a paramagnetic atom.
[0227] Preferred paramagnetic atoms are divalent or trivalent ions
of elements with an atomic number of 21 to 29, 42, 44 and 58 to 70.
Suitable ions include chromium(III), manganese(II), iron(III),
iron(II), cobalt(II), nickel(II), copper(II), praseodymium(III),
neodymium(III), samarium(III) and ytterbium(III). Because of their
very strong magnetic moments, gadolinium(III), terbium(III),
dysoprosium(III), holmium(III), and erbium(III) are preferred.
Especially preferred for the paramagnetic atom is
gadolinium(III).
[0228] The compositions ofthe present invention may be prepared by
combining a compound of Formula I with a paramagnetic atom. For
example, the metal oxide or a metal salt (for example, nitrate,
chloride or sulfate) of a suitable paramagnetic atom is dissolved
or suspended in a medium comprised of water and an alcohol, such as
methyl, ethyl or isopropyl alcohol. This mixture is added to a
solution of an equimolar amount of the compound of Formula I in a
similar aqueous medium and stirred. The reaction mixture may be
heated moderately until the reaction is completed. Insoluble
compositions formed may be isolated by filtering, while soluble
compositions may be isolated by evaporation of the solvent. If acid
groups on the chelating means are still present in the composition
of the present invention, inorganic or organic bases, and even
amino acids, may be added to convert the acidic complex into a
neutral complex to facilitate isolation or purification of
homogenous composition. Organic bases or basic amino acids may be
used as neutralizing agents, as well as inorganic bases such as
hydroxides, carbonates or bicarbonates of sodium, potassium or
lithium.
[0229] The present invention also include diagnostic compositions
which are useful for in vivo imaging of thrombi in a mammal,
comprising a pharmaceutically acceptable carrier and a
diagnostically effective amount of compositions derived from the
compounds of Formula I. Compositions such as those described in
paragraphs B and C herein above may be conveniently used in these
diagnostic compositions.
[0230] The "diagnostically effective amount" of the composition
required as a dose will depend on the route of administration, the
type of mammal being treated, and the physical characteristics of
the specific mammal under consideration. These factors and their
relationship to determining this dose are well known to skilled
practitioners in the medial diagnostic arts. Also, the
diagnostically effective amount and method of administration can be
tailored to achieve optimal efficacy but will depend on such
factors as weight, diet, concurrent medication and other factors
which those skilled in the medical arts will recognize. In any
regard, the dose for imaging should be sufficient for detecting the
presence of the imaging agent at the site of a thrombus in
question. Typically, radiologic imaging will require that the dose
provided by the pharmaceutical composition position of the present
invention be about 5 to 20 .mu.Ci, preferably about 10 .mu.Ci.
Magnetic resonance imaging will require that the dose provided be
about 0.001 to 5 mmole/kg, preferably about 0.005 to 0.5 mmole/kg
of a compound of Formula VII complexed with paramagnetic atom. In
either case, it is known in the art that the actual dose will
depend on the location of the thrombus.
[0231] "Pharmaceutically acceptable carriers" for in vivo use are
well known in the pharmaceutical art, and are described, for
example, in Remington's Pharmaceutical Sciences, Mack Publishing
Co. (A. R. Gennaro edit. 1985). The pharmaceutical compositions of
the present invention may be formulated with a pharmaceutically
acceptable carrier to provide sterile solutions or suspensions for
injectable administration. In particular, injectables can be
prepared in conventional forms, either as liquid solutions or
suspensions, solid forms suitable for solution or suspensions in
liquid prior to injection, or as emulsions. Suitable excipients
are, for example, water, saline, dextrose, mannitol, lactose,
lecithin, albumin, sodium glutamate, cysteine hydrochloride, or the
like. In addition, if desired, the injectable pharmaceutical
compositions may contain minor amounts of nontoxic auxiliary
substances, such as wetting agents, pH buffering agents, and the
like. If desired, absorption enhancing preparations (e.g.,
liposomes) may be utilized.
[0232] The present invention also encompasses diagnostic
compositions prepared for storage or administration. These would
additionally contain preservatives, stabilizers and dyes. For
example, sodium benzoate, sorbic acid and esters of
p-hydroxybenzoic acid may be added as preservatives. Id. At 1449.
In addition, antioxidants and suspending agents may be used.
Id.
[0233] The in vivo imaging methods of the present invention also
offer several advantages over previous imaging techniques for the
detection or monitoring of the presence, size, regression or
increase of a thrombus. In particular, the present invention
provides compounds, compositions and diagnostic compositions have
been designed to bind extremely tightly to the thrombin associated
with a thrombus and thereby reduce "background" due to circulating
radioactivity or paramagnetism arising from unbound imaging agent.
Furthermore, in vivo imaging by intracoronary injection ofthe
compounds, compositions or diagnostic compositions of the present
invention, is expected to be almost instantaneous since these
imaging agents would saturate the thrombin bound to the thrombus
immediately.
[0234] Accordingly, the present invention also includes methods for
in vivo imaging of a thrombus in a mammal, comprising the steps of:
(1) administering to a mammal a diagnostically acceptable amount of
a compound, composition, or diagnostic composition of the present
invention and (2) detecting a thrombus in a blood vessel.
[0235] The term "in vivo imaging" as used herein relates to methods
of the detection of a thrombus in a meal, as well as the monitoring
of the size, location and number of thrombi in a mammal, as well as
dissolution or growth of the thrombus.
[0236] In employing the compounds, compositions or diagnostic
compositions in vivo by this method, "administering" is
accomplished parenterally, in either a systemic or local targeted
manner. Systemic administration is accomplished by injecting the
compounds, compositions by diagnostic compositions ofthe present
invention into a convenient and accessible vein or artery. This
includes but is not limited to administration by the ankecubutal
vein. Local targeted administration is accomplished by injecting
the compounds, compositions or diagnostic compositions of the
present invention proximal in flow to a vein or artery suspected to
contain thrombi distal to the injection site. This includes but is
not limited to direct injection into the coronary arterial
vasculature to image coronary thrombi, into the carotid artery to
image thrombi in the cerebral vasculature, or into a pedal vein to
image deep vein thrombosis of the leg.
[0237] Also, the manner of delivery of a composition of the present
invention to the site of a thrombus is considered within the scope
of the term "administering". For example, a compound represented by
Formula I having a chelating means attached thereto may be injected
into the mammal, followed at a later time by the radioactive atom
thereby forming in vivo at the site of the thrombus the composition
comprising the compound of formula complexed to radioactive atom.
Alternatively, a composition comprising the compound of formula
complexed to radioactive atom may be injected into the mammal.
[0238] The "diagnostically effective amount" of the compounds,
compositions or diagnostic compositions used in the methods of the
present invention will, as previously mentioned, depend on the
route ofadministration, the type ofmammal being treated, and the
physical characteristics of the specific mammal under treatment.
These factors and their relationship to determining this dose are
well known to skilled practitioners in the medical diagnostic arts.
In any regard, the dose for in vivo imaging should be sufficient
for detecting the presence of the imaging agent at the site of a
thrombus in question. Typically, radiologic imaging will require
that the dose provided by the diagnostic composition of the present
invention be about 5 to 20 .mu.Ci, preferably about 10 .mu.Ci.
Magnetic resonance imaging will require that the dose provided by
the diagnostic composition be about 0.001 to 5 mmole/kg, preferably
about 0.005 to 0.5 mmole/kg of a compound of Formula I complexed
with paramagnetic atom. In either case it is known in the art that
the actual dose will depend on the location of the thrombus.
[0239] The detecting of a thrombus by imaging is made possible by
the presence of radioactive or paramagnetic atoms localized at such
thrombus.
[0240] The radioactive atoms associated with the compositions and
diagnostic compositions of the present invention are preferably
imaged using a radiation detection means capable of detecting gamma
radiation, such as a gamma camera or the like. Typically, radiation
imaging cameras employ a conversion medium (wherein the high energy
gamma ray is absorbed, displacing an electron which emits a photon
upon its return to the orbital state), photoelectric detectors
arranged in a spatial detection chamber (to determine the position
of the emitted photons), and circuitry to analyze the photons
detected in the chamber and produce an image.
[0241] The paramagnetic atoms associated with the compositions and
diagnostic compositions of the present invention detected in
magnetic resonance imaging (MRI) systems. In such systems, a strong
magnetic field is used to align the nuclear spin vectors ofthe
atoms in apatients body. The field is disturbed by the presence of
paramagnetic atoms localized at a thrombus and an image of the
patient is read as the nuclei return to their equilibrium
alignments.
[0242] The following examples are illustrative, but not limiting,
of the method and compositions of the present invention. Other
suitable modifications and adaptations ofthe variety of conditions
and parameters normally encountered and obvious to those skilled in
the art are within the spirit and scope of the invention. 27 28
EXAMPLE 1
[3-[5-Chloro-3-(N-cyclopentyl-N-[prop-2-enyl]aminocarbonyl)
phenoxy}propoxyamino]carboxamidine hydrochloride (21)
[0243] Compound numbers appearing in Examples 1-21 refer to
compounds having the structures shown in Schemes 3 and 4. 29
[0244] a.) 1,3-Diclhloro-5-(tert-butyldimethylsilyloxy)benzene
(13)
[0245] To a solution of CH.sub.2Cl.sub.2 (60 mL) and
3,5-dichlorophenol (5.0 g, 30 miol) was added
tert-butyldimethylsilyl chloride (5.54 g, 36 mmor ,
N,N-diisopropylethylamine (8.0 mL, 46 mmol) and a catalytic amount
of 4-dimethylaminopyridine. The initially exothermic solution was
stirred at ambient temperature for 6 h then diluted with
CH.sub.2Cl.sub.2 (40 mL). The mixture was washed consecutively with
10% aqueous HCl (50 mL), saturated aqueous NaHCO.sub.3 (50 mL), and
brine (50 mL). The organic phase was dried over anhydrous
MgSO.sub.4, filtered and concentrated in vacuo to provide 13 as a
pale yellow liquid (8.8 g. 100%). .sup.1H NMR (300 MHz, CDCl.sub.3)
d 6.98 (s, 1 H), 6.72 (s, 2 H), 0.98 (s, 9 H), 0.22 (s, 6 H).
[0246] b.) 3-Chloro-5-(tert-butyldimethylsilyloxy)benzoic acid
(14)
[0247] To "Rieke Mg" (0.21 mol; Rieke, R. D.; Bales, S. E.;
Hudnall, P. M.; Bums, T. P.; Poindexter, G. S. Org. Synth.
Collective Volume VI, 1988, 845.) in tetrahydrofuran (1000 niL) was
added ether 13 (27.7 g, 0.10 mol). After the reaction mixture was
stirred for 20 min at ambient temperature there was an exotherm
observed. The exotherm subsided within 5 min, and the reaction
mixture was cooled to 20.degree. C. with an ice bath. After 15 min,
the reaction mixture was cooled to -78.degree. C. To the cool
reaction mixture was added CO.sub.2 gas for 30 min. The reaction
mixture was warmed to ambient temperature, then diluted with cold
(0.degree. C.) 10% aqueous HCl (150 mL) and ethyl acetate (400 mL).
The aqueous phase was extracted with ethyl acetate (400 mL). The
combined organic phases were washed with brine, dried over
anhydrous MgSO.sub.4, filtered and concentrated in vacuo. The
resulting solid was recrystallized from acetonitrile to provide 14
as fluffy white needles (19.3 g, 64%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.7.70 (s, 1 H), 7.43 (s, 1 H), 7.08 (s, 1 H),
0.99 (s, 9 H), 0.23 (s, 6H). IR (KBr) 2957, 1697, 1578, 1434, 1297,
1266, 1115, 990, 871 cm.sup.-1.
[0248] c.)
[3-chloro-5-(tert-butyldimethylsilyloxy)phenyl]-N-cyclopentyl-N-
-prop-2-enylearboxamide (15)
[0249] To a solution of CH.sub.2Cl.sub.2 (250 mL) and acid 14 (17.5
g, 60 mmol) was added triethylamine (33.8 mL, 0.24 mol) and
bis(2-oxo-3-oxazolidinyl)phosphinic chloride (17.0 g, 66 mmol). The
resulting mixture was stirred for 5 min, then
N-allylcyclopentylamine (9.8 mL, 66 mmol) was added. The mixture
was stirred for 1 h then filtered. The filtrate was washed with 10%
aqueous HCl (100 mL), saturated aqueous NaHCO.sub.3 (100 mL) and
brine (100 mL). The organic layer was dried over anhydrous
MgSO.sub.4 and concentrated in vacuo. The crude product was
purified by chromatography on silica gel to provide compound 15 as
a colorless oil (23.5 g, 97%). .sup.1H NMR (300 MHz. CDCl.sub.3)
.delta.6.95 (s, 1 H), 6.84 (s, 1 H), 6.71 (s, 1 H), 5.93 (bs, 1 H),
5.16 (d, 2 H). 3.95 (bs, 3 H), 1.4-1.9 (m, 8 H), 0.97 (s, 9 H),
0.20 (s, 6 H). Mass spectrum (CI) calcd. for
C.sub.21H.sub.32NO.sub.2SiCl- : 394 (M+H). Found: 394.
[0250] d.)
(5-chloro-3-lydroxyphenyl)-N-cyclopentyl-N-prop-2-enylcarboxami- de
(16)
[0251] To a solution of compound 15 (23.4 g, 59 mmol) and
tetrahydrofuran (200 mL) was added tetrabutyl-ammonium fluoride (1
M intetrahydrofaran, 66 mL, 66 mmol). The solution was stirred for
30 min, then poured into a separatory finnel containing 10% aqueous
HCl (100 mL) and ethyl acetate (200 mL). The layers were separated
and the aqueous layer was extracted with ethyl acetate (200 mL).
The organic layers were combined, washed with brine (100 mL), dried
over anhydrous MgSO.sub.4 and concentrated in vacuo. The crude
product was purified by chromatography on silica gel to yield
phenol 16 (12 g, 72%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta.8.73 (bs, 1 H), 6.82 (s, 2 H), 6.76 (s, 1 H), 5.95 (bs, 1
H), 5.16-5.23 (m, 2 H), 3.7-4.15 (m, 3 H), 1.45-2.0 (m, 8 H). IR
(NaCI) 3177, 2956, 1590, 1433, 1373, 1289, 935 cm.sup.-1.
[0252] e.)
[3-chloro-5-(3-hydroxypropoxy)phenyl]-N-cyclopentyl-N-prop-2-en- yl
carboxamide (17)
[0253] To a solution of tetrahydrofuran (100 mL), 1,3-propanediol
(1.6 mL, 30 mmol) and phenol 16 (4.3 g, 15 mmol) at 0.degree. C.,
was added diethyl azodicarboxylate (DEAD, 3.6 mL, 23 mmol) dropwise
over 10 min. The solution was warmed to ambient temperature and
stirred for 16 h, prior to concentration in vacuo. The oil was
partially purified by chromatography on silica gel to provide
alcohol 17 (3.9 g) contaminated with Mitsunobu by-products. This
material was used directly in subsequent experiments. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta.6.93 (s, 2 H), 6.8 (s, 1 H), 5.9 (bs,
1 H), 5.18 (d, 2 H), 4.4-3.8 (m, 7 H), 2.05 (m, 2 H), 1.4-1.9 (m,
10 H). Mass spectrum (CI) calcd. for C.sub.18H.sub.24NO.sub.3- Cl:
338 (M+H). Found: 338.
[0254] f.)
{5-[3-(1,3-dioxoisoindolin-2-yloxy)propoxy]-3-chlorophenyl}-N-c-
yclopentyl-N-prop-2-enylcarboxamide (18)
[0255] A mixture of alcohol 17 (8.8 g, 26 mmol), tetrahydrofuran
(100 mL), N-hydroxyphthalimide (3.8 g, 23 mmol) and
triphenylphosphine (5.2 g, 20 mmol) was stirred at 0.degree. C. for
5 min. To the mixture was added DEAD (3.4 mL. 20 mmol) dropwise.
The reaction mixture was then allowed to warm to ambient
temperature and stirred for 12 h prior to concentration in vacuo.
The residue was purified by chromatography to provide phthalimide
18 (3.7 g) contaminated with Mitsunobu by-products. This material
was used directly in subsequent experiments. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.7.78-7.85 (m, 2 H), 7.73-7.77 (m, 2 H), 6.94 (t,
2 H), 6.38 (d, 1 H), 5.95 (bs, 1 H), 5.18 (d, 2 H), 4.40 (t, 2 H),
3.8-4.4 (m, 4 H), 2.20-2.28 (m, 2 H), 1.4-1.9 (m, 1 H). Mass
spectrum (CI) calcd. for C.sub.26H.sub.27N.sub.2O.sub.5Cl: 483
(M+H). Found: 483.
[0256] g.)
{5-[3-(aminooxy)propoxy]-3-chlorophenyl}-N-cyclopentyl-N-prop-2-
-enyl carboxamide (19)
[0257] To a solution of ethanol (50 mL) and phthalimide 18 (1.8 g,
3.7 mmol) was added 40% aqueous methylamine (25 mL). The reaction
mixture was stirred at ambient temperature for 16 h. The reaction
mixture was concentrated in vacuo and then chromatographed on
silica gel to provide hydroxylamine 19 (0.5 g; 15% from 16).
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta.6.92 (s, 2 H), 6.79 (s, 1
H), 5.92 (bs, 1 H), 5.40 (s, 2 H), 5.17 (m, 2 H), 4.04 (t, 2 H),
3.88-4.03 (m, 3 H), 3.82 (t, 2 H), 2.01-2.12 (m, 2 H), 1.4-1.9 (m,
8 H).
[0258] h.)
[3-{5-Chloro-3-(N-cyclopentyl-N-[prop-2-enyl]aminocarbonyl)phen-
oxy}propoxyaminolcarboxamidine (20)
[0259] To a solution of amine 19 (1.4 g, 4 mmol) in N,
N-dimethylformamide (20 mL) was added 1 H-pyrazole-1-carboxamidine
hydrochloride (0.61 g, 4 mmol). The reaction mixture was stirred
for 18 h, concentrated in vacuo, and purified by chromatography on
silica gel to provide 20 (0.94 g, 61%) as a viscous oil.
[0260] i.)
[3-{5-Chloro-3-(N-cyclopentyl-N-[prop-2-enyl]aminocarbonyl)phen-
ox}propoxyamino]carboxamidine hydrocliloride (21)
[0261] To a solution of 20 (0.6 g, 1.5 mmol) and ethyl acetate (10
mL) was added 4N HCl in ethyl acetate (4 mL). The resulting mixture
was concentrated to dryness in vacuo, and triturated with ether to
provide title compound 21 (0.57 g, 88%) as a white amorphous solid.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta.7.07 (s, 1 H), 6.95 (s, 1
H), 6.85 (s, 1 H), 5.93 (bs, 1 H), 5.18 (d, 2 H), 4.14 (t, 2 H),
4.07 (t, 2 H), 3.95-4.1 (m, 2 H). .sup.13C NMR (CD.sub.3OD) 8
161.0, 136.0, 119.6, 116.9, 112.3, 75.2, 66.3,30.7,28.7,25.0.
IR(KBr)3355, 3137, 2951, 1670, 1618, 1437, 1052, 651 cm.sup.-1.
Mass spectrum (CI) calcd. for C.sub.19H.sub.27N.sub.4O.sub.3Cl: 395
(M+H). Found: 395.
EXAMPLE 2
[0262]
[3-{5-Chloro-3-(4-benzylpiperidinylcarbonyl)phenoxy}propoxyamino]ca-
rboxamidine trifluoroacetate (32) 30
[0263] a.) Benzyl3-chloro-5-(tert-butyldimetliylsilyloxy)benzoate
(24)
[0264] To a mixture of the acid 14 (3.80 g, 13.3 mmol) and oxalyl
chloride (5.8 mL, 66 mmol) in methylene chloride (60 mL) was added
ca. 0.05 mL of N,N-dimethylformamide as a catalyst. After stirring
1 hr at ambient temperature, the reaction was evaporated in vacuo,
diluted with methylene chloride, and reacted with benzyl alcohol
(1.38 mL, 13.3 mmol) and 4-(N,N-dimethylamino)pyridine (1.60 g,
13.3 mmol). After stirring 16 hrs at ambient temperature, the
solution was washed with dilute aqueous HCl, dilute aqueous
NaHCO.sub.3, and brine, dried over Na.sub.2SO.sub.4, and filtered.
The filtrate was then evaporated in vacuo giving a quantitative
yield of gold oil. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta.7.64
(t, 1H, J=1.5 Hz), 7.45-7.28 (m, 6 H), 7.02 (m, 1 H), 5.35 (s, 2
H), 0.98 (s, 9 H), 0.22 (s, 6 H).
[0265] b.) Benzyl 3-ckloro-5-hydroxybenzoate (25)
[0266] A 1.0 M solution oftetrabutylammonium fluoride in
tetrahydrofuiran (26.6 mL, 26.6 mmol) was added to neat benzyl
ester 24 (5.00 g, 13.3 mmol), the reaction stirred 10 minutes at
ambient temperature, and evaporated in vacuo. The crude product was
dissolved in methylene chloride, the solution washed with dilute
aqueous NaHCO.sub.3, dried over Na.sub.2SO.sub.4, and filtered. The
evaporated filtrate was then washed repeatedly with hexanes and
dried in vacuo giving product 25 as a gold oil (3.40 g, 98%).
H.sup.1NMR (300 MHz, CDCl.sub.3) .delta.7.35 (m, 7 H), 7.1 (t, 1 H,
J=2.1 Hz), 5.28 (s, 2 H).
[0267] c.) Benzyl3-chloro-5-(3-hydroxypropoxy)benzoate (26)
[0268] To a solution of phenol 25 (8.80 g, 33.0 mmol) and 3-bromo-
1-propanol (2.9 mL, 33 mmol) in acetonitrile (300 mL) was added
solid cesium carbonate (12 g, 37 mmol). After stirring 16 hrs at
50.degree. C., more 3-bromo- I -propanol (3.3 mmol) and cesium
carbonate (3.3 mmol) were added and the reaction stirred another 2
hrs. After adding more 3-bromo-1-propanol and cesium carbonate (3.3
mmol each) and sodium iodide (3.3 mmol), the reaction was stirred
30 minutes at 65.degree. C., then cooled and filtered. The filtrate
was evaporated in vacuo, the residue dissolved in ethyl acetate,
washed with water, and the organic layer dried over
Na.sub.2SO.sub.4 and filtered. The evaporated filtrate then gave
product 26 (5.96 g. 56%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta.7.63 (t, 1 H, J=1.5 Hz), 7.48 (dd, 1 H, J=2.4 Hz, 1.4 Hz),
7.46-7.31 (m, 5 H), 7.09 (t, IH, J=2.4 Hz), 5.35 (s, 2 H), 4.14 (t,
2 H, J=6.0 Hz), 3.85 (t, 2 H, J=6.0 Hz), 2.05 (pentet, 2 H, J=6.0
Hz).
[0269] d.) Benzyl
5-[3-(1,3-dioxoisoindolin-2-yloxy)propoxy]-3-chlorobenzo- ate
(27)
[0270] A solution of alcohol 26 (5.96 g, 18.6 mmol),
triphenylphosphine (5.40 g, 20.5 mmol), and N-hydroxyphthalimide
(2.10 g, 20.5 mmol) in tetrahydrofuran (186 mL) was cooled to
0.degree. C. and reacted slowly with diethyl azodicarboxylate (3.50
mL, 22.3 mmol). After warming to ambient temperature and stirring
16 hrs, the reaction was concentrated in vacuo and purified by
chromatography on silica gel (10% ethyl acetate in hexanes) giving
product 27 (5.18 g, 60%). .sup.1H NMR (300 MHz, CDCI.sub.3)
.delta.7.83 (dd, 2 H, J=5.6 Hz, 3.1 Hz), 7.75 (dd, 2 H, J=5.6 Hz,
3.1 Hz), 7.63 (t, 1 H, J=1.5 Hz), 7.51 (dd, 1 H, J =2.4 Hz, 1.4
Hz), 7.40 (m, 5 H), 7.12 (t, 1 H, J=2.3 Hz), 5.35 (s, 2 H), 4.41
(t, 2 H, J=6.1 Hz), 4.28 (t, 2 H, J=6.1 Hz), 2.25 (pentet, 2 H,
J=6.1 Hz).
[0271] e.) Benzyl 5-[3-(aminooxy)propoxy]-3-chlorobenzoate (28)
[0272] To a solution of phthalimide 27 (1.00 g, 2.15 mmol) in
tetrahydrofuran (20 mL) was added a 40% aqueous solution of
methylamine (0.148 mL, 1.72 mmol). After stirring 25 minutes at
0.degree. C., the reaction was evaporated in vacuo, the residue
partitioned between methylene chloride and water, and the organic
layer dried over Na.sub.2SO.sub.4 and filtered. The evaporated
filtrate was then purified by chromatography on silica gel (40%
ethyl acetate in hexanes) giving product 28 (0.576 g,80%). .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta.7.62 (t, 1 H, J =1.6 Hz), 7.48
(dd, 1 H, J=2.4 Hz, 1.4 Hz), 7.45-7.32 (m, 5H), 7.08 (t, lH, J =2.1
Hz), 5.35 (s, 2 H), 4.07 (t, 2 H, J=6.3 Hz), 3.82 (t, 2 H, J=6.3
Hz), 2.07 (pentet, 2 H, J =6.3 Hz).
[0273] f) Benzyl
5-[3-{N,N'-di-(tert-butoxycarbonyl)amidinoaminooxy}propoy-
]-3-chlorobenzoate (29)
[0274] A solution of amine 28 (0.710 g, 2.10 mmol) and
N,N'-bis(tert-butoxycarbonyl)amidinopyrazole (0.724 g, 2.30 mmol)
in N,N-dimethylformamide (10 mL) was stirred for 3 days at ambient
temperature and evaporated in vacuo. The crude product was purified
by chromatography on silica gel (20% diethyl ether in petroleum
ether) giving compound 29 (0.83 g, 69%). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.9.08 (s, 1 H), 7.70 (s, 1 H), 7.62 (t, 1 H,
J=1.6 Hz), 7.47 (dd, 1 H, J=2.5 Hz, 1.4Hz), 7.45-7.32 (m, 5 H),
7.09 (t, 1 H, J=2.1 Hz), 5.35 (s, 2 H), 4.23 (t, 2 H, J=6.0 Hz),
4.10 (t, 2 H, J =6.0 Hz), 2.18 (pentet, 2 H, J=6.2 Hz), 1.49 (s, 18
H).
[0275] g.)
5-[3-N,N'-di-(tert-butoxycarbonyl)amindinoaminooxy}propoxy]-3-c-
hlorobenzoic acid (30)
[0276] To a solution of product 29 (2.80 g, 4.85 mmol) in ethanol
(48 mL) was added a 2 N aqueous solution of NaOH (22 mL, 44 mmol).
The solution was stirred 30 minutes atambienttemperature andthe
ethanol removed in vacuo. The remaining solution was acidified to
pH 3 with 10% aqueous citric acid and extracted with methylene
chloride. The organic layer was dried over Na.sub.2SO.sub.4,
filtered, the filtrate evaporated, and the crude product purified
by chromatography on silica gel (30% ethyl acetate in hexanes)
giving compound 30 (1.50 g, 64%). .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta.9.10 (s, 1 H), 8.73 (bs, 1 H), 7.69 (bs, IH), 7.66 (t, 1 H,
J=1.5 Hz), 7.50 (dd, 1 H, J=2.3 Hz, 1.3 Hz), 7.12 (t, 1 H, J=2.3
Hz), 4.24 (t, 2 H, J=6.0 Hz), 4.12 (t, 2 H, J=6.2 Hz), 2.20
(pentet, 2 H, J=6.1 Hz), 1.50 (s, 18 H). Mass spectrum (LCMS, ESI
pos.) calcd. for C.sub.21H.sub.30N.sub.3O.sub.8Cl: 288.0 (M-2
Boc+H). Found: 288.2.
[0277] h.)
[3-{5-Chloro-3-(4-benzylpiperidinylcarbonyl)phenoxy}propoxyamin-
o]-N,N'-di-(tert-butoxycarbonyl)carboxamidine (31)
[0278] A solution of product 30 (0.032 g, 0.066 mmol),
benzotriazole-1-yl-oxy-tris-(dimethylamino)-phosphonium
hexafluorophosphate (0.032 g, 0.072 mmol), and triethylamine (0.01
mL, 0.07 mmol) in methylene chloride (1.0 mL) was added to a vial
containing diethylaminomethyl-polystyrene resin (0.06 g) and
4-benzylpiperidine (0.014 g, 0.077 mmol). The reaction was shaken
for 3 days, poured onto a Waters 2 g silica Sep-Pak, and the
product eluted with 5 to 15% methanol in methylene chloride. The
product-containing fractions were combined, washed with 10% aqueous
citric acid and saturated aqueous NaHCO.sub.3, and dried over
Na.sub.2SO.sub.4. The decanted solution was evaporated in vacuo
giving a clear oil that was used directly in the proceeding step
without analysis.
[0279] i)
[3-{5-chloro-3-(4-benzylpiperidinylcarbonyl)phenoxy}propoxyamino-
]carboxamidine trifluoroacetate (32)
[0280] Product 31 was dissolved in methylene chloride (2.0 mL),
treated with trifluoroacetic acid (0.5 mL), tightly capped, and
shaken for 18 hrs at ambient temperature. The solution was
evaporated in vacuo and the crude product purified on a Waters 2 g
silica Sep-Pak (5 to 20% methanol in methylene chloride) giving
title compound 32 (0.032 g, 87% from 30). .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta.7.33-7.19 (m, 3 H), 7.15 (m, 2 H), 6.96 (m, 1
H), 6.92 (t, 1 H, J=1.5 Hz), 6.79 (dd, 1 H, 2.3 Hz, 1.3 Hz), 4.60
(bd, 1 H, 13 Hz), 4.10 (t, 2 H, J=5.9Hz), 4.07 (t, 2 H, J=6.1 Hz),
2.98 (bt, 1 H, J=12 Hz), 2.73 (bt, IH, J=13 Hz), 2.58 (d. 2 H,
J=7.0 Hz), 2.15 (pentet, 2 H, J=6.0 Hz), 1.83 (m, 2 H), 1.65 (bd,
lH, J=13 Hz), 1.21 (m, 2 H). Mass spectrum (MALDI-TOF, gentisic
acid matrix) calcd. for C.sub.23 H.sub.29N.sub.4O.sub.3CI: 445.2
(M+H). Found: 445.1.
EXAMPLE 3
[0281]
[3-{5-Chloro-3-(N,N-bis[2-metlioxyethyl]aminocarbonyl)phenoxy}propo-
xyamnino]carboxamidine trifluoroacetate 31
[0282] The title compound was prepared from compound 30 and
bis(2-methoxyethyl)amine in a manner analogous to steps h and i of
Example 2. .sup.1H NMR (300 MHz, CDCl.sub.3/CD.sub.3OD) .delta.7.00
(t, 1 H, J=1.6 Hz), 6.95 (t, 1 H, J=2.1 Hz), 6.86 (dd, 1 H, J=2.3
Hz, 1.3 Hz), 4.10 (t, 2 H, J=5.8 Hz), 4.07 (t, 2 H, J=6.1 Hz), 3.72
(bm, 2 H), 3.66 (bm, 2 H), 3.52 (bm, 2 H), 3.46 (bm, 2 H), 3.39
(bs, 3 H), 3.30 (bs, 3 H), 2.15 (pentet, 2 H, J=6.2 Hz). Mass
spectrum (MALDI-TOF, gentisic acid matrix) calcd. for C.sub.17
H.sub.27N.sub.4O.sub.5Cl: 402.2 (M). Found: 402.0.
EXAMPLE 4
[0283]
[3-{5-Chloro-3-(N-methyl-N-[2-{2-pyridyl}ethyl]aminocarbonyl)plleno-
xy}propoxyaminolcarboxamidine trifluoroacetate 32
[0284] The title compound was prepared from compound 30 and
methyl(2-pyridyl-2-ethyl)amine in a manner analogous to steps h and
i of Example 2. .sup.1H NMR (300 MHz, CDCl.sub.3/CD.sub.3OD)
.delta.8.70 (m, 1 H), 8.18 (t, 1 H, J=7.6 Hz), 7.75 (d, 1 H, J=7.9
Hz), 7.64 (bt, 1 H, J=6.5 Hz), 6.95 (bs, lH), 6.86 (bs, 1 H), 6.74
(bs, 1 H), 4.08 (m, 6 H), 3.90 (t, 2 H, J=6.8 Hz), 3.37 (m, 2 H),
3.02 (s, 3 H), 2.14 (pentet, 2 H, J=5.9 Hz). Mass spectrum
(MALDI-TOF, gentisic acid matrix) calcd. for C.sub.19
H.sub.24N.sub.5O.sub.3Cl: 406.2 (M+H). Found: 406.3.
EXAMPLE 5
[0285]
[3-{5-Chloro-3-(N-methyl-N-[3-pyridylmethyl]aminocarbonyl)phenoxy}p-
ropoxyamino]carboxamidine trifluoroacetate 33
[0286] The title compound was prepared from compound 30 and
methyl(3-pyridylmethyl)amine in a manner analogous to steps h and i
of Example 2. .sup.1H NMR(300 MHz, CDCl.sub.3/CD.sub.3OD)
.delta.8.74-8.65 (bm, IH), 8.11 (bm, 1 H), 7.67 (bm, 1 H), 7.53
(bm, 1 H), 6.99 (m, 2 H), 6.88 (bs, 1 H), 4.80 (s, 2 H), 4.08 (m, 4
H), 3.00 (bs, 3 H), 2.16 (m, 2 H). Mass spectrum (MALDI-TOF,
.alpha.-cyano-4-hydroxycinnamic acid matrix) calcd. for
C.sub.18H.sub.22N.sub.5O.sub.3Cl: 391.1 (M). Found: 391.5.
EXAMPLE 6
[0287]
[3-{5-Chloro-3-(N-ethyl-N-[4-pyridylmethyl]aminocarbonyl)phenoxy}pr-
opoxyamino]carboxamidine trifluoroacetate 34
[0288] The title compound was prepared from compound 30 and
ethyl(4-pyridylmethyl)amine in a manner analogous to steps h and i
of Example 2. Mass spectrum (MALDI-TOF, gentisic acid matrix)
calcd. for C.sub.19H.sub.24N.sub.5O.sub.3Cl: 405.2 (M). Found:
405.5.
EXAMPLE 7
[0289] Ethyl
2-[5-{3-(amidinoaminooxy)propoxy}-3-chlorophenyl]-N-{2-pyridy-
lmethyl}aminocarbonyl]acetate trifluoroacetate 35
[0290] The title compound was prepared from compound 30 and
2-pyridylmethylglycine ethyl ester in a manner analogous to steps h
and i of Example 2. Mass spectrum (MALDI-TOF, gentisic acid matrix)
calcd. for C.sub.21H.sub.26N.sub.5O.sub.5Cl: 463.2 (M). Found:
463.5.
EXAMPLE 8
[0291]
[3-{5-Chloro-3-([2-{3,4-dihydroxyphenyl}-2-oxoethyl]-N-methyl
aminocarbonyl)phenoxy}propoxyamino]carboxamidine trifluoroacetate
36
[0292] The title compound was prepared from compound 30 and
methyl-2-(3,4-dihydroxyphenylacetyl)amine in a manner analogous to
steps h and i of Example 2. Mass spectrum (MALDI-TOF, gentisic acid
matrix) calcd. for C.sub.20H.sub.23N.sub.4O.sub.6Cl: 450.1 (M).
Found: 450.2.
EXAMPLE 9
[0293]
[3-{5-Chloro-3-(N-[2-{dimethlylamino}ethyl]-N-ethylaminocarbonyl)ph-
enoxy}propoxyamino]carboxamidine trifluoroacetate 37
[0294] The title compound was prepared from compound 30 and
ethyl(2-dimethylaminoethyl)amine in a manner analogous to steps h
and i of Example 2. .sup.1H NMR (300 MHz, CDCl.sub.3/CD.sub.3OD)
.delta.6.99 (t, 1 H, J=2.1 Hz), 6.96 (m, 1 H), 6.88 (bs, 1 H), 4.10
(m, 4 H), 3.83 (t, 2 H, J=6.6 Hz), 3.38 (m, 4 H). 96 (bs, 6 H),
2.16 (pentet, 2 H, J=6.0 Hz), 1.14 (t, 3 H, J=7.0 Hz). Mass
spectrum (MALDI-TOF, .alpha.-cyano-4-hydroxycinnamic acid matrix)
calcd. for C.sub.17H.sub.28N.sub.5O.sub.3Cl: 385.2 (M). Found:
385.6.
EXAMPLE 10
[0295]
[3-{5-Chloro-3-(4-formylpiperazinylcarbonyl)phenoxy}propoxyamino]ca-
rboxamidine trifluoroacetate 38
[0296] The title compound was prepared from compound 30 and
4-piperazinecarboxaldehyde in a manner analogous to steps h and i
of Example 2. .sup.1H NMR (300 MHz, CDC.sub.3/CD.sub.3OD)
.delta.8.08 (bs, 1 H), 7.01 (t, 1 H, J=2.1 Hz), 6.97 (m, 1 H), 6.84
(dd, 1 H, J=2.3 Hz, 1.4 Hz), 4.14-4.06 (m, 4 H), 2.16 (pentet, 2 H,
6.0 Hz). Mass spectrum (MALDI-TOF, gentisic acid matrix) calcd. for
C.sub.16H.sub.22N.sub.5O.sub- .4Cl: 383.1 (M). Found: 383.2.
EXAMPLE 11
[0297]
3-{5-Chloro-3-(4-phenylpiperazinylcarbonyl)phenoxy}propoxyamino]car-
boxamidine trifluoroacetate 39
[0298] The title compound was prepared from compound 30 and
4-phenylpiperazine in a manner analogous to steps h and i of
Example 2. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.7.27-6.85 (m, 8
H), 4.16 (t, 2 H, J=6.1 Hz), 4.08 (t, 2 H, J=6.3 Hz), 3.31 (m, 8
H), 2.18 (pentet, 2 H, J=6.2 Hz). Mass spectrum (LCMS, ESI pos.)
calcd. for C.sub.21H.sub.26N.sub.5O.sub.3Cl: 432.2 (M+H). Found:
432.3.
EXAMPLE 12
[0299] [3-{5-Chloro-3-(4-benzilpiperazinylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate 40
[0300] The title compound was prepared from compound 30 and
4-benzylpiperazine in a manner analogous to steps h and i of
Example 2. Mass spectrum (LCMS, ESI pos.) calcd. for
C.sub.22H.sub.28N.sub.5O.sub.3C- l: 446.2 (M+H). Found: 446.6.
EXAMPLE 13
[0301] [3-{5-Chloro-3-(N,N-dimethylaminocarbonyl)
phenoxy}propoxyamino]car- boxamidine trifluoroacetate 41
[0302] The title compound was prepared from compound 30 and
N,N-dimethylamine in a manner analogous to steps h and i of Example
2. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.10.99 (s, 1 H), 7.64
(bs, 4 H), 7.10 (t, 1 H J=2.0 Hz), 7.02 (m, 1 H), 6.91 (m, 1 H),
4.14 (t, 2 H, J=6.3 Hz), 3.94 (t, 2 H, J=6.3 Hz), 2.96 (s, 3 H),
2.88 (s, 3 H), 2.06 (m, 2 H). Mass spectrum (LCMS, ESI pos.) calcd.
for C.sub.13H.sub.19N.sub.4O.sub.3Cl: 315.1 (M+H). Found:
315.4.
EXAMPLE 14
[0303]
[3-{5-Chloro-3-(piperidinylcarbonyl)phenoxy}propoxyaminolcarboxamid-
ine trifluoroacetate 42
[0304] The title compound was prepared from compound 30 and
piperidine in a manneranalogous to steps hand i of Example 2.
.sup.1H NMR(300 MHz, DMSO-d.sub.6) .delta.7.67 (bs, 4 H), 7.10 (t,
1 H, J=2.0 Hz), 6.98 (t, 1 H, J=2.0 Hz), 6.87 (dd, 1 H, J=2.4Hz,
1.3 Hz), 4.14 (t, 2 H, J=6.2 Hz), 3.94 (t, 2 H, J=6.4 Hz), 2.06
(pentet, 2 H, J=6.3 Hz), 1.60-1.24 (m, 6 H). Mass spectrum (LCMS,
ESI pos.) calcd. for C.sub.16H.sub.23N.sub.4O.su- b.3Cl: 355.1
(M+H). Found: 355.3.
EXAMPLE 15
[0305] [3-{5-Chloro-3-(4-[2-pyridyl]piperazinylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate 43
[0306] The title compound was prepared from compound 30 and
4-(2-pyridyl)piperazine in a manner analogous to steps h and i of
Example 2. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.11.19 (s, 1
H), 8.11 (m, 1 H), 7.77 (bs, 3 H), 7.62 (m, 1 H), 7.14 (t, 1 H,
J=2.1 Hz), 7.08 (t, 1 H, J=1.5 Hz), 6.97 (dd, 1 H, J=2.3 Hz, 1.3
Hz), 6.90 (d, 1 H, J=8.7 Hz), 6.71 (dd, 1 H, J=6.8 Hz, 5.2 Hz),
4.15 (t, 2 H, J=6.4 Hz), 3.95 (t, 2 H, J=6.4 Hz), 3.70-3.42 (bm, 8
H), 2.07 (pentet, 2 H, J=6.3 Hz). Mass spectrum (LCMS, ESI pos.)
calcd. for C.sub.20 H.sub.25N.sub.6O.sub.3Cl: 433.2 (M+H). Found:
431.6.
EXAMPLE 16
[0307] [3-{5-Chloro-3-(2-[1,2,3,4-tetrahydro]isoquinolinylcarbonyl)
phenoxy}propoxyamino]carboxamidine trifluoroacetate 44
[0308] The title compound was prepared from compound 30 and
1,2,3,4-tetrahydroisoquinoline in a manner analogous to steps h and
i of Example 2. Mass spectrum (LCMS, ESI pos.) calcd. for C.sub.20
H.sub.23N.sub.4O.sub.3Cl: 403.1 (M+H). Found: 403.3.
EXAMPLE 17
[0309] [3-{5-Chloro-3-(azaperhydroepinylcarbonyl)
phenoxy}propoxyamino]car- boxamidine trifluoroacetate 45
[0310] The title compound was prepared from compound 30 and
N,N-cyclohexylimine in a manner analogous to steps h and i of
Example 2. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.11.23 (s, 1
H), 7.79 (bs, 4 H), 7.08 (t, 1 H, J=2.1 Hz), 6.97 (m, 1 H), 6.86
(dd, 1 H, J=2.3 Hz, 1.3 Hz), 4.13 (t, 2 H, J=6.3 Hz), 3.94 (t, 2 H,
J=6.4 Hz), 3.53 (t, 2 H, J=5.8 Hz), 3.27 (m, 2 H), 2.08 (m, 2 H),
1.71 (bm, 2 H), 1.53 (bm, 6 H). Mass spectrum (LCMS, ESI pos.)
calcd. for C.sub.17H.sub.25N.sub.4O.sub.3C- l: 369.2 (M+H). Found:
369.3.
EXAMPLE 18
[0311] Ethyl
3-({5-[3-(amidinoaminooxy)propoxy]-3-chlorophenyl}-N-benzylca-
rbonylamino)propanoate trifluoroacetate 46
[0312] The title compound was prepared from compound 30 and benzyl
(3-ethyl propionato)amine in a manner analogous to steps h and i of
Example 2. Mass spectrum (LCMS, ESI pos.) calcd. for
C.sub.23H.sub.29N.sub.4O.sub.5Cl: 477.2 (M+H). Found: 477.4.
EXAMPLE 19
[0313] Ethyl
1-({5-[3-(amidinoaminooxy)propoxy]-3-chlorophetiyljcarbonyl)p-
iperidine-4-carboxylate trifluoroacetate 47
[0314] The title compound was prepared from compound 30 and
4-(carboxyethyl)piperidine in a manner analogous to steps h and i
of Example 2. .sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta.11.26 (s,
1 H), 7.81 (bs, 4 H), 7.11 (t, 1 H, J=2.1 Hz), 7.00 (t, 1 H,
J=1.6Hz), 6.90 (dd, 1 H, J=2.3 Hz, 1.2 Hz), 4.13 (t, 2 H, J=6.3
Hz), 4.08 (q, 2 H, J=7.1 Hz), 3.94 (t, 2 H, J 6.4 Hz), 3.09-2.89
(bm, 2 H), 2.63 (tt, 1 H, J=10.9 Hz, 3.8 Hz), 2.07 (pentet, 2 H,
J=6.3 Hz), 1.89 (bm, 2 H), 1.52 (bm, 2 H), 1.18 (t, 3 H, J=7.1 Hz).
Mass spectrum (LCMS, ESI pos.) calcd. for
C.sub.19H.sub.27N.sub.4O.sub.5Cl: 427.2 (M+H). Found: 427.3.
EXAMPLE 20
[0315] [3-{5-Chloro-3-(morpholin-4-ylcarbonyl)
phenoxy}propoxyamino]carbox- amidine trifluoroacetate 48
[0316] The title compound was prepared from compound 30 and
morpholine in 15 a manner analogous to steps hand i of Example 2.
.sup.1H NMR (300 MHz. DMSO-d.sub.6) .delta.11.25 (s, 1 H), 7.80
(bs, 4 H), 7.12 (t, 1 H, J=2.1 Hz), 7.03 (t. 1 H. J=1.6 Hz), 6.93
(dd, 1 H. J=2.3 Hz, 1.2 Hz), 4.14 (t, 2 H, J=6.3 Hz), 3.94 (t. 2 H,
J=6.4 Hz), 3.59 (bm, 6 H), 3.31 (bm, 2 H), 2.07 (pentet, 2 H, J=6.3
Hz). Mass spectrum (LCMS, ESI pos.) calcd. for
C.sub.15H.sub.21N.sub.4O.sub.4Cl: 357.2 (M+H). Found: 357.6.
EXAMPLE 21
[0317] Methyl
2-({5-[3-(amidinoaminooxt)propxy]-3-chlorophenyl}-N-methtylc-
arbonylamino)acetate trifluoroacetate 49
[0318] The title compound was prepared from compound 30 and
methyl(2-methyl acetonato)amine in a manner analogous to steps h
and i of Example 2. Mass spectrum (LCMS, ESI pos.) calcd. for
C.sub.15H.sub.21N.sub.4O.sub.5Cl: 373.2 (M+H). Found: 373.5.
EXAMPLE 22
[0319] Tablet Preparation
[0320] Tablets containing 25.0, 50.0, and 100.0 mg, respectively,
of the active compound (compound 21 of Example 1) are prepared as
illustrated below:
1 TABLET FOR DOSES CONTAINING FROM 25-100 MG OF THE ACTIVE COMPOUND
Amount-mg Active Compound 25.0 50.0 100.00 Microcrystalline
cellulose 37.25 100.0 200.0 Modified food corn starch 37.25 4.25
8.5 Magnesium stearate 0.50 0.75 1.5
[0321] All of the active compound, cellulose, and a portion of the
corn starch are mixed and granulated to 10% corn starch paste. The
resulting granulation is sieved, dried and blended with the
remainder of the corn starch and the magnesium stearate. The
resulting granulation is then compressed into tablets containing
25.0, 50.0, and 100.0 mg, respectively, of active ingredient per
tablet.
EXAMPLE 23
[0322] Intravenous Solution Preparation
[0323] An intravenous dosage form of the above-indicated active
compounds is prepared as follows:
2 Active Compound 0.5-10.0 mg Sodium Citrate 5-50 mg Citric Acid
1-15 mg Sodium Chloride 1-8 mg Water for Injection (USP) q.s. to 1
ml
[0324] Utilizing the above quantities, the active compound (21) is
dissolved at room temperature in a previously prepared solution of
sodium chloride, citric acid, and sodium citrate in Water for
Injection (USP, see page 1636 of United States
Pharmacopeia/National Formulary for 1995, published by United
States Pharmacopeial Convention, Inc., Rockville, Md. (1994).
EXAMPLE 24
[0325] In vitro Inhibition of Purified Enzymes
[0326] Reagents: All buffer salts were obtained from Sigma Chemical
Company (St. Louis, Mo.), and were of the highest purity available.
The enzyme substrates, N-benzoyl-Phe-Val-Arg-p-nitroanilide (Sigma
B7632), N-benzoyl-Ile-Glu-Gly-Arg- p-nitroanilide hydrochloride
(Sigma B2291), N-p-Tosyl-Gly-Pro-Lys-p-nitroanilide (Sigma T6140),
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (Sigma S7388)
andN-CBZ-Val-Gly-Arg-p-nitroanilide (Sigma C7271) were obtained
from Sigma. N-succinyl-Ala-Ala-Pro-Arg-p-nitroanilide (BACHEM
L-1720) and N-succinyl-Ala-Ala-Pro-Val-p-nitroanilide (BACHEM
L-1770) were obtained from BIACHEM (King of Prussia, Pa.).
[0327] Human .alpha.-thrombin, human factor Xa and human plasmin
were obtained from Enzyme Research Laboratories (South Bend,
Indiana). Bovine .alpha.-chymotrypsin (Sigma C4129), bovine trypsin
(Sigma T8642) and human kidney cell urokinase (Sigma U5004) were
obtained from Sigma. Human leukocyte elastase was obtained from
Elastin Products (Pacific, Mo.).
[0328] K.sub.i Determinations: All assays are based on the ability
of the test compound to inhibit the enzyme catalyzed hydrolysis of
a peptidep-nitroanilide substrate. In a typical K.sub.i
determination, substrate is prepared in DMSO, and diluted into an
assay buffer consisting of 50 mM HEPES, 200 mM NaCl, pH 7.5. The
final concentrations for each of the substrates is listed below. In
general, substrate concentrations are lower than the experimentally
determined value for K.sub.m. Test compounds are prepared as a 1.0
mg/ml solution in DMSO. Dilutions are prepared in DMSO yielding 8
final concentrations encompassing a 200 fold concentration range.
Enzyme solutions are prepared at the concentrations listed below in
assay buffer.
[0329] In a typical K.sub.i determination, into each well of a 96
well plate is pipetted 280 mL of substrate solution, 10 mL of test
compound solution, and the plate allowed to thermally equilibrate
at 37.degree. C. in a Molecular Devices plate reader for >15
minutes. Reactions were initiated by the addition of a 10 mL
aliquot of enzyme and the absorbance increase at 405 nm is recorded
for 15 minutes. Data corresponding to less than 10% of the total
substrate hydrolysis were used in the calculations. The ratio of
the velocity (rate of change in absorbance as a function of time)
for a sample containing no test compound is divided by the velocity
of a sample containing test compound, and is plotted as a function
of test compound concentration. The data are fit to a linear
regression, and the value of the slope of the line calculated. The
inverse of the slope is the experimentally determined K.sub.i
value.
[0330] Thrombin: Thrombin activity was assessed as the ability to
hydrolyze the substrate N-succinyl-Ala-Ala-Pro-Arg-p-nitroanilide.
Substrate solutions were prepared at a concentration of 32 mM
(32mM<<Km=180 mM) in assay buffer. Final DMSO concentration
was 4.3%. Purified human a-thrombin was diluted into assay buffer
to a concentration of 15 riM. Final reagent concentrations were:
[thrombin] =0.5 nM,
[substrateN-succinyl-Ala-Ala-Pro-Arg-p-nitroanilide]=32 mM.
[0331] Factor X [FXa]: FXa activity was assessed as the ability to
hydrolyze the substrate N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide
hydrochloride. Substrate solutions were prepared at a concentration
of 51 mM (51<<K.sub.m=1.3 mM) in assay buffer. Final DMSO
concentration was 4.3%. Purified activated human Factor X was
diluted into assay buffer to a concentration of 300 nM. Final
reagent concentrations were: [FXa]=10 nM,
[N-benzoyl-Ile-Glu-Gly-Arg-p-nitroanilide hydrochloride]=51 mM.
[0332] Plasmin: Plasmin activity was assessed as the ability to
hydrolyze the N-p-Tosyl-Gly-Pro-Lys-p-nitroanilide. Substrate
solutions were prepared at a concentration of 37 mM (37
mM<<K.sub.m=243 mM) in assay buffer. Final DMSO concentration
was 4.3%. Purified human plasmin was diluted into assay buffer to a
concentration of 240 nM. Final reagent concentrations were:
[Plasmin]=8 nM, [N-p-Tosyl-Gly-Pro-Lys-p-nitroanilid- e]=37 mM.
[0333] Chymotrypsin: Chymotrypsin activity was assessed as the
ability to hydrolyze N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide.
Substrate solutions were prepared at a concentration of 14 mM (14
mM<<K.sub.m=62 mM) in assay buffer. Final DMSO concentration
was 4.3%. Purified bovine chymotrypsin was diluted into assay
buffer to a concentration of 81 nM. Final reagent concentrations
were: [Chymotrypsin]=2.7 nM,
[N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide]=14 mM.
[0334] Trypsin: Trypsin activity was assessed as the ability to
hydrolyze N-benzoyl-Phe-Val-Arg-p-nitroanilide. Substrate solutions
were prepared at a concentration of 13 mM (13 mM<<K.sub.m=291
mM) in assay buffer. Final DMSO concentration was 4.3%. Purified
bovine trypsin was diluted into assay buffer to a concentration of
120 nM. Final reagent concentrations were: [Trypsin]=4 nM,
[N-benzoyl-Phe-Val-Arg-p-nitroanilid- e]=13 mM.
[0335] Elastase: Elastase activity was assessed as the ability to
hydrolyze N-succinyl-Ala-Ala-Pro-Val-p-nitroanilide. Substrate
solutions were prepared at a concentration of 19 mM (19
mM<<K.sub.m=89 mM) in assay buffer. Final DMSO concentration
was 4.3%. Purified human leukocyte elastase was diluted into assay
buffer to a concentration of 750 nM. Final reagent concentrations
were: [Elastase]=25 nM,
[N-succinyl-Ala-Ala-Pro-Val-p-nitroanilide]=19 mM.
[0336] Urokinase: Urokinase activity was assessed as the ability to
hydrolyze N-CBZ-Val-Gly-Arg-p-nitroanilide. Substrate solutions
were prepared at a concentration of 100 mM (100 mM<K=1.2 mM) in
assay buffer. Final DMSO concentration was 4.3%. Purified human
kidney urokinase was diluted into assay buffer to a concentration
of 1.2 mM. Final reagent concentrations were: [Urokinase]=40 nM,
and N-CBZ-Val-Gly-Arg-p-nitroanilide]=100 mM.
[0337] The exemplified compounds of the present invention had K's
for thrombin between 20 nM and 12 .mu.M.
[0338] The results indicate that the compounds of the present
invention are inhibitors of proteases, including thrombin.
[0339] Having now fully described this invention, it will be
understood to those of ordinary skill in the art that the same can
be performed within a wide and equivalent range of conditions,
formulations, and other parameters without affecting the scope of
the invention or any embodiment thereof. All patents and
publications cited herein are fully incorporated by reference
herein in their entirety.
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