Note: Descriptions are shown in the official language in which they were submitted.
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SULFONYLATED DIPEPTIDE COMPOUNDS WHICH INHIBIT
LEUKOCYTE ADHESION MEDIATED BY VLA-4
BACKGROUND OF THE INVENTION
Field of the Invention
This invention relates to compounds which inhibit leukocyte adhesion
and, in particular, leukocyte adhesion mediated by VLA-4.
rences
The following publications, patents and patent applications are cited
in this application as superscript numbers:
' Hemler and Takada, European Patent Application Publication
No. 330,506, published August 30, 1989
Elices, et al., Cell, ø0_:577-584 (1990)
3 Springer, Nature, x:425-434 (1990)
Osborn, Cell, ~:3-6 (1990)
Vedder, et al., Surgery,11~.:509 (1989)
Pretolani, et al., J. Exp. Med.,1$Q:795 (1994)
' Abraham, et al., J. Clin. Invest., x.:776 (1994)
g Mulligan, et al., J. Immunology, 5]~Q:2407 (1993)
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Cybulsky, et al., Science, X15 :788 (1991)
' Li, et al., Arterioscler. Thromb., 1:197
(1993)
" Sasseville, et al., Am. J. Path., 14:27
(1994)
'z Yang, et al., Proc. Nat. Acad. Science
(USA), 9:10494
(1993)
'3 Burkly, et al., Diabetes, x:529 (1994)
Baron, et al., J. Clin. Invest., x:1700
(1994)
Hamann, et al., J. Immunology, .L52:3238
(1994)
Yednock, et al., Nature, ~5 :63 (1992)
" Baron, et al., J. Exp. Med., X77:57 (1993)
'8 van Dinther-Janssen, et al., J. Immunology,
x:4207 (1991)
van Dinther-Janssen, et al., Annals. Rheumatic
Dis., x:672
( 1993)
z Elices, et al., J. Clin. Invest., ,3:405
(1994)
z' Postigo, et al., J. Clin. Invest., $x:1445
(1991)
'-'- Paul, et al., Transpl. Proceed., 2:813
(1993)
Okarhara, et al., Can. Res., x:3233 (1994)
za paavonen, et al., Int. J. Can., x:298 (1994)
zs Schadendorf, et al., J. Path., x:429 (
1993)
z6 Bao, et al., Diff., ,5:239 (1993)
z' Lauri, et al., British J. Cancer, $:862
(1993)
z8 Kawaguchi, et al., Japanese J. Cancer Res.,
$x:1304 (1992)
z9 Kogan, et al., U.S. Patent No. 5, 510,
332, issued April 23,
1996
3 International Patent Appl. Publication
No. WO 96/01644
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All of the above publications, patents and patent applications are
herein incorporated by reference in their entirety to the same extent as if
each
individual publication, patent or patent application was specifically and
individually indicated to be incorporated by reference in its entirety.
State of the Art
VLA-4 (also referred to as a4(3, integrin and CD49d/CD29), first
identified by Hemler and Takada' is a member of the p I integrin family of
cell surface receptors, each of which comprises two subunits, an a chain and
a ~i chain. VLA-4 contains an a4 chain and a ~i 1 chain. There are at least
nine X31 integrins, all sharing the same ~i 1 chain and each having a distinct
a
chain. These nine receptors all bind a different complement of the various
cell matrix molecules, such as fibronectin, laminin, and collagen. VLA-4,
for example, binds to fibronectin. VLA-4 also binds non-matrix molecules
i5 that are expressed by endothelial and other cells. These non-matrix
molecules include VCAM-1, which is expressed on cytokine-activated
human umbilical vein endothelial cells in culture. Distinct epitopes of VLA-
4 are responsible for the fibronectin and VCAM-1 binding activities and each
activity has been shown to be inhibited independently.Z
Intercellular adhesion mediated by VLA-4 and other cell surface
receptors is associated with a number of inflammatory responses. At the site
of an injury or other inflammatory stimulus, activated vascular endothelial
cells express molecules that are adhesive for leukocytes. The mechanics of
leukocyte adhesion to endothelial cells involves, in part, the recognition and
binding of cell surface receptors on leukocytes to the corresponding cell
surface molecules on endothelial cells. Once bound, the leukocytes migrate
across the blood vessel wall to enter the injured site and release chemical
mediators to combat infection. For reviews of adhesion receptors of the
immune system, see, for example, Springer3 and Osborn4.
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Inflammatory brain disorders, such as experimental autoimmune
encephalomyelitis (EAE), multiple sclerosis {MS) and meningitis, are
examples of central nervous system disorders in which the
endothelium/leukocyte adhesion mechanism results in destruction to
otherwise healthy brain tissue. Large numbers of leukocytes migrate across
the blood brain barrier (BBB) in subjects with these inflammatory diseases.
The leukocytes release toxic mediators that cause extensive tissue damage
resulting in impaired nerve conduction and paralysis.
In other organ systems, tissue damage also occurs via an adhesion
mechanism resulting in migration or activation of leukocytes. For example,
it has been shown that the initial insult following myocardial ischemia to
heart tissue can be further complicated by leukocyte entry to the injured
tissue causing still further insult (Vedder et a1.5). Other inflammatory
conditions mediated by an adhesion mechanism include, by way of example,
asthmab'8, Alzheimer's disease, atherosclerosis9''°, AIDS dementia",
diabetes'2'" (including acute juvenile onset diabetis), inflammatory bowel
disease's (including ulcerative colitis and Crohn's disease), multiple
sclerosis'6'", rheumatoid arthritis'8'2', tissue transplantation2z, tumor
metastasis23-is, meningitis, encephalitis, stroke, and other cerebral traumas,
nephritis, retinitis, atopic dermatitis, psoriasis, myocardial ischemia and
acute leukocyte-mediated lung injury such as that which occurs in adult
respiratory distress syndrome.
In view of the above, assays for determining the VLA-4 level in a
biological sample containing VLA-4 would be useful, for example, to
diagnosis VLA-4 mediated conditions. Additionally, despite these advances
in the understanding of leukocyte adhesion, the art has only recently
addressed the use of inhibitors of adhesion in the treatment of inflammatory
brain diseases and other inflammatory conditionsz9,so. The present invention
addresses these and other needs.
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SUMMARY OF THE INVENTION
This invention provides compounds which bind to VLA-4. Such
compounds can be used, for example, to assay for the presence of VLA-4 in
a sample and, in pharmaceutical compositions, to inhibit cellular adhesion
5 mediated by VLA-4, for example, binding of VCAM-1 to VLA-4. The
compounds of this invention have a binding affinity to VLA-4 as expressed
by an ICso of about 15 ~cM or less (as measured using the procedure shown in
Example 203 below} which compounds are defined by formula I below:
R3 O
II
R'-SOZ-N(RZ)-C-Q-CH-C-OH
H RS
where
R' is selected from the group consisting of alkyl, substituted alkyl,
aryl, substituted aryl, cycioalkyl, substituted cycloalkyl, heterocyclic,
substituted heterocylic, heteroaryl and substituted heteroaryl;
R'- is selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,
substituted
cycloalkenyl, heterocyclic, substituted heterocyclic, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, and R' and RZ together with the nitrogen
atom bound to Rz and the SOZ group bound to R' can form a heterocyclic or a
substituted heterocyclic group;
R3 is selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic and
where Rz and R3 together with the nitrogen atom bound to Rz and the carbon
atom bound to R' can form a saturated heterocyclic group or a saturated
substituted heterocyclic group with the proviso that when monosubstituted,
the substituent on said saturated substituted heterocyclic group is not
carboxyl;
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Ar is aryl, heteroaryl, substituted aryl or substituted heteroaryl;
x is an integer of from 1 to 4;
Q is -C(X)NR'- wherein R' is selected from the group consisting of
hydrogen and alkyl;
X is selected from the group consisting of oxygen and sulfur;
RS is -CHZX where X is selected from the group consisting of
hydrogen, hydroxyl, acylamino, alkyl, alkoxy, aryloxy, aryl, aryloxyaryl,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, cycloalkyl, substituted alkyl, substituted
alkoxy, substituted aryl, substituted aryloxy, substituted aryloxyaryl,
substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic;
with the further provisos that:
A. R5 is not selected from the group consisting of -(CHZ)~ aryl
and -(CHZ)"heteroaryl where n is an integer equal to 1 to 4 when RZ and R3
together with the nitrogen atom bound to Rz and the carbon atom bound to R3
form a saturated heterocyclic group or a saturated substituted heterocyclic
group;
B. RS is not -(CHZ)X Ar-R5~ where RS' is selected from the group
consisting of -O-Z-NRBR$' and -O-Z-R'2 wherein Ar is aryl, heteroaryl,
substituted aryl or substituted heteroaryl, x is an integer of from 1 to 4, R8
and R8~ are independently selected from the group consisting of hydrogen,
alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic,
and
substituted heterocyclic, and where R8 and Rg' are joined to form a
heterocycle or a substituted heterocycle, R'2 is selected from the group
consisting of heterocycles and substituted heterocycles, and Z is selected
from the group consisting of -C(O)- and -SOZ-;
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C. R5 is not -(CHz)X-Ar-RS' where Ar is aryl, substituted aryl,
heteroaryl or substituted heteroaryl, x is an integer of from 1 to 4, R5' is
selected from the group consisting of -NRzaC(Z')NRgRg~ and
-NRZaC{Z')R" wherein Z' is selected from the group consisting of oxygen,
sulfur and NRZa, RZa is selected from the group consisting of hydrogen, alkyl
and aryl, R8 and Rg' are independently selected from the group consisting of
hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl,
substituted cycloalkyl, heterocyclic, substituted heterocyclic, heteroaryl and
substituted heteroaryi provided that when Z' is oxygen, at least one of R8 and
R8' is sustituted alkyl, cycloalkyl, substituted cycloalkyl, saturated
heterocyclic other than morpholino and thiomorpholino, substituted
heterocyclic or R$ and Rg' are joined to form a saturated heterocycle other
than morpholino or thiomorpholino, a saturated substituted heterocycle or a
saturated/unsaturated heterocycle having an amino group substituted with an
alkoxycarbonyl substituent, and further provided that when Z' is sulfur, at
least one of R$ and R8' is a group other than aryl, substituted aryl,
heteroaryl
or substituted heteroaryl, and R'3 is selected from the group consisting of
substituted heterocyles and saturated heterocycles other than morpholino and
thiomorpholino;
D. RS is not -ALK-X where ALK is an alkyl group of from 1 to
10 carbon atoms attached via a methylene group (-CHI-) to the carbon atom
to which it is attached; X is selected from the group consisting of
substituted
alkylcarbonylamino, substituted alkenylcarbonylamino, substituted
alkynylcarbonylamino, heterocyclylcarbonylamino, substituted
heterocyclylcarbonylamino, acyl, acyloxy, aminocarbonyloxy, acylamino,
oxycarbonylamino, alkoxycarbonyl, substituted alkoxycarbonyl,
aryloxycarbonyl, substituted aryloxycarbonyl, cycloalkoxycarbonyl,
substituted cycloalkoxycarbonyl, heteroaryloxycarbonyl, substituted
heteroaryloxycarbonyl, heterocyclyloxycarbonyl, substituted
heterocyclyloxycarbonyl, cycloalkyl, substituted cycloalkyl, saturated
heterocyclic, substituted saturated heterocyclic, substituted alkoxy,
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g
substituted alkenoxy, substituted alkynoxy, heterocyclyloxy, substituted
heterocycloxy, substituted thioalkyl, substituted thioalkenyl, substituted
thioalkynyl, aminocarbonylamino, aminothiocarbonylamino, guanidino,
amidino, alkylamidino, thioamidino, halogen, cyano, vitro, -OS(O)2-alkyl,
-OS(O)2-substituted alkyl, -OS(O)Z-cycloalkyl, -OS(O)Z-substituted
cycloalkyl, -OS{O)2-aryl, -OS(O)2-substituted aryl, -OS(O)Z-heteroaryl,
-OS(O)2-substituted heteroaryl, -OS(O)z-heterocyclic, -OS(O)2-substituted
heterocyclic, -OSOZ-NRR where R is hydrogen or alkyl, -NRS(O)2-alkyl,
-NRS{O)2-substituted alkyl, -NRS(O)z-cycloalkyl, -NRS{O)2-substituted
cycloalkyl, -NRS(O)2-aryl, -NRS(O)2-substituted aryl, -NRS(O)z-heteroaryl,
-NRS{O)2-substituted heteroaryl, -NRS(O)2-heterocyclic, -NRS(O)z-
substituted heterocyclic, -NRS(O)2-NR-alkyl, -NRS(O)2-NR-substituted
alkyl, -NRS(O)2-NR-cycloalkyl, -NRS(O)2-NR-substituted cycloalkyl,
-NRS(O),-NR-aryl, -NRS(O)Z-NR-substituted aryl, -NRS(O)z-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O),-NR-heterocyclic,
-NRS(O)2-NR-substituted heterocyclic where R is hydrogen or alkyl, -S(O)~-
alkyl, -S(O)2-substituted alkyl, -S(O)S-aryl, -S(O),-substituted aryl, -S(O)Z-
substituted heteroaryl, -S(O)2-substituted heteroaryl, -S(O)S-heterocyclic,
-S(O)Z-substituted heterocyclic, mono- and di-(substituted alkyl)amino, N,N-
(alkyl, substituted alkyl)amino, N,N-(aryl, substituted alkyl)amino, N,N-
(substituted aryl, substituted alkyl)amino, N,N-(heteroaryl, substituted
alkyl)amino, N,N-(substituted heteroaryl, substituted alkyl)amino, N,N-
(heterocyclic, substituted alkyl)amino, N,N-N,N-(substituted heterocyclic,
substituted alkyl)amino, mono- and di-(heterocyclic)amino, mono- and di-
(substituted heterocyclic)amino, N,N-(alkyl, heterocyclic)amino, N,N-{alkyl,
substituted heterocyclic)amino, N,N-(aryl, heterocyclic)amino, N,N-
(substituted aryl, heterocyclic)amino, N,N-(aryl, substituted
heterocyclic)amino, N,N-(substituted aryl, substituted heterocyclic)amino,
N,N-(heteroaryl, heterocyclic)amino, N,N-(heteroaryl, substituted
heterocyclic)amino, N,N-(substituted heteroaryl, heterocyclic)amino, and
N,N-(substituted heteroaryl, substituted heterocyclic)amino; and
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E. RS is not -(CHz)X Ar-RS" where RS" is a substituent selected
from the group consisting of:
(a) substituted alkylcarbonylamino with the proviso that at least one
of the substituents on the substituted alkyl moiety is selected from the group
consisting of alkoxy, substituted alkoxy, acyl, acylamino,
thiocarbonylamino, acyloxy, alkenyl, amino, amidino, alkyl amidino,
thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryloxy, substituted aryloxy, cyano, nitro, halogen,
hydroxyl; carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-
cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-
substituted aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,
substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl,
substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,
thioheterocyclic, substituted thioheterocyclic, heterocyclic, substituted
heterocyclic, cycloalkoxy, substituted cycloalkyoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -OS(O)Z-alkyl, -OS(O)2-
substituted alkyl, -OS(O)z-aryl, -OS(O)z-substituted aryl, -OS(O)Z-heteroaryl,
-OS(O)2-substituted heteroaryl, -OS(O)2-heterocyclic, -OS(O)2-substituted
heterocyclic, -OSOZ-NRR, -NRS(O)2-alkyl, -NRS(O)Z-substituted alkyl, -
NRS(O)Z-aryl, -NRS(O)2-substituted aryl, -NRS(O)z-heteroaryl, -NRS(O)z-
substituted heteroaryl, -NRS(O)Z-heterocyclic,
-NRS(O),-substituted heterocyclic, -NRS(O)2-NR-alkyl, -NRS(O)Z-NR-
substituted alkyl, -NRS(O)~-NR-aryl, -NRS(O)Z-NR-substituted aryl,
-NRS(O)2-NR-heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O)z-
NR-heterocyclic, -NRS(O)Z-NR-substituted heterocyclic, mono- and di-
alkylamino, mono- and di-(substituted alkyl)amino, mono- and di-arylamino,
mono- and di-(substituted aryl)amino, mono- and di-heteroarylamino, mono-
and di-(substituted heteroaryl)amino, mono- and di-heterocyclic amino,
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mono- and di-(substituted heterocyclic) amino, unsymmetric di-substituted
amines having different substituents selected from the group consisting of
alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, heterocyclic and substituted heterocyclic, substituted alkyl
groups
5 having amino groups blocked by conventional blocking groups (such as Boc,
Cbz, formyl, and the like), and alkyl/substituted alkyl groups substituted
with
-SOz-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SOz-substituted alkenyl,
-SOZ-cycloalkyl, -SOZ-substituted cycloalkyl, -SOZ-aryl, -SOZ-substituted
aryl, -SO,-heteroaryl, -SOZ-substituted heteroaryl, -SOS-heterocyclic, -SOZ-
10 substituted heterocyclic or -SOZNRR, where R is hydrogen or alkyl;
(b) alkoxyaryl substituted on the alkoxy moiety with a substituent
selected from the group consisting of carboxyl and -COORz3 where R23 is
alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic,
(c) aryl and heteroaryl;
(d) -NR'R' wherein each R' is independently selected from the group
consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,
substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic and
substituted heterocyclic with the proviso that at least one of R' is
substituted
alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic and substituted
heterocyclic and with the further proviso that when R' is substituted alkyl at
least one of the substituents on the substituted alkyl moiety is selected from
the group consisting of alkoxy, substituted alkoxy, acyl, acylamino,
thiocarbonylamino, acyloxy, alkenyl, amino, amidino, alkyl amidino,
thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryloxy, substituted aryloxy, cyano, nitro, halogen,
hydroxyl, carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-
cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-
substituted aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,
substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl,
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substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,
thioheterocyclic, substituted thioheterocyclic, heterocyclic, substituted
heterocyclic, cycloalkoxy, substituted cycloalkyoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -OS(O)2-alkyl, -OS(O)2-
substituted alkyl, -OS(O)2-aryl, -OS(O)2-substituted aryl, -OS(O)2-heteroaryl,
-OS(O)2-substituted heteroaryl, -OS(O)2-heterocyclic, -OS(O)Z-substituted
heterocyclic, -OS02-NRR, -NRS(O)2-alkyl, -NRS(O)2-substituted alkyl,
-NRS(O)2-aryl, -NRS(O)Z-substituted aryl, -NRS(O)Z-heteroaryl, -NRS(O}z-
substituted heteroaryl, -NRS(O)2-heterocyclic, -NRS(O)z-substituted
heterocyclic, -NRS(O)2-NR-alkyl, -NRS(O)2-NR-substituted alkyl,
-NRS(O)2-NR-aryl, -NRS(O)2-NR-substituted aryl, -NRS(O)Z-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O)2-NR-heterocyclic,
-NRS(O)z-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
i5 di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, and unsymmetric di-substituted amines
having different substituents, wherein the substituents are selected from the
group consisting of alkyl, substituted alkyl, aryl, substituted aryl,
heteroaryl,
substituted heteroaryl, heterocyclic and substituted heterocyclic, substituted
alkyl groups having amino groups blocked by conventional blocking groups
(such as Boc, Cbz, formyl, and the like), and alkyl/substituted alkyl groups
substituted with -SOZ-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SOz-
substituted alkenyl, -SOZ-cycloalkyl, -SOZ-substituted cycloalkyl, -SOZ-aryl,
-SOZ-substituted aryl, -SOZ-heteroaryl, -SOZ-substituted heteroaryl, -SOZ-
heterocyclic, -SOZ-substituted heterocyclic or -SOZNRR, where R is
hydrogen or alkyl;
(e) -alkoxy-NR"R" wherein each R" is independently selected from
the group consisting of alkyl, substituted alkyl, aryl, substituted aryl,
cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl,
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heterocyclic, and substituted heterocyclic with the proviso that when each R"
is substituted alkyl then at least one of the substituents on the substituted
alkyl moiety is selected from the group consisting of alkoxy, substituted
alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, alkenyl, amino,
amidino, alkyl amidino, thioamidino, aminoacyl, aminocarbonylamino,
aminothiocarbonylamino, aminocarbonyloxy, aryloxy, substituted aryloxy,
cyano, vitro, halogen, hydroxyl, carboxyl, carboxylalkyi, carboxyl-
substituted alkyl, carboxyl-cycIoalkyl, carboxyl-substituted cycloalkyl,
carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-
substituted heteroaryl, carboxylheterocyclic, carboxyl-substituted
heterocyclic, cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone,
thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,
thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substituted
thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heterocyclic,
substituted heterocyclic, cycloalkoxy, substituted cycloalkyoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -OS(O)2-alkyl, -
OS(O)2-substituted alkyl, -OS(O)2-aryl, -OS(O)2-substituted aryl, -OS(O)z-
heteroaryl, -OS(O)Z-substituted heteroaryl, -OS(O)Z-heterocyclic, -OS(O)2-
substituted heterocyclic, -OSO,-NRR, -NRS(O)z-alkyl, -NRS(O)2-substituted
alkyl, -NRS(O)2-aryl, -NRS(O)2-substituted aryl, -NRS(O)~-heteroaryl, -
NRS(O)2-substituted heteroaryl, -NRS(O)2-heterocyclic, -NRS(O)2-
substituted heterocyclic, -NRS(O)z-NR-alkyl, -NRS(O)2-NR-substituted
alkyl, -NRS(O)2-NR-aryl, -NRS(O)2-NR-substituted aryl, -NRS(O)2-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O)2-NR-heterocyclic,
-NRS(O)Z-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl;
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substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocycIic, substituted alkyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like), and alkyl/substituted alkyl groups substituted with -
SOZ-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SOZ-substituted alkenyl, -
SOZ-cycloalkyl, -SOZ-substituted cycloalkyl, -S02-aryl, -SOZ-substituted aryl,
-SOZ-heteroaryl, -SOZ-substituted heteroaryl, -SO~-heterocyclic, -SOz-
substituted heterocyclic or -SOZNRR, where R is hydrogen or alkyl;
(f) substituted alkenyl or substituted alkynyl with the proviso that at
least one of the substituents on the substituted alkenyl/alkynyl moiety is
selected from the group consisting of alkyl, substituted alkyl, aryl,
substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted
heteroaryl, heterocyclic, and substituted heterocyclic with the proviso that
when substituted with substituted alkyl, then at least one of the substituents
on the substituted alkyl moiety is selected from the group consisting of
alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy,
alkenyl, amino, amidino, alkyl amidino, thioamidino, aminoacyl,
aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aryloxy,
substituted aryloxy, cyano, nitro, halogen, hydroxyl, carboxyl, carboxylalkyl,
carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substituted
cycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,
carboxyl-substituted heteroaryl, carboxylheterocyclic, carboxyl-substituted
heterocyclic, cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone,
thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,
thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substituted
thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heterocyclic,
substituted heterocyclic, cycloalkoxy, substituted cycloalkyoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -OS(O),-alkyl, -
OS(O)z-substituted alkyl, -OS(O),-aryl, -OS(O)~-substituted aryl, -OS(O),-
heteroaryl, -OS(O),-substituted heteroaryl, -OS(O),-heterocyclic, -OS(O),-
*rB
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substituted heterocyclic, -OSOZ-NRR, -NRS(O)2-alkyl, -NRS(O)2-substituted
alkyl, -NRS(O)2-aryl, -NRS(O)2-substituted aryl, -NRS(O),-heteroaryl, -
NRS(O)Z-substituted heteroaryl, -NRS(O)Z-heterocyclic, -NRS(O)2-
substituted heterocyclic, -NRS(O)2-NR-alkyl, -NRS{O)2-NR-substituted
alkyl, -NRS(O)Z-NR-aryl, -NRS(O)2-NR-substituted aryl, -NRS(O)2-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O)Z-NR-heterocyclic,
-NRS(O),-NR-substituted heterocyclic or mono- and di-alkylamino, mono-
and di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic, substituted alkyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like) and alkyl/substituted alkyl groups substituted with -
SO,-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SO~-substituted alkenyl, -
SOZ-cycloalkyl, -SO~-substituted cycloalkyl, -SO,-aryl, -SO,-substituted aryl,
-SOz-heteroaryl, -SO,-substituted heteroaryl, -SO,-heterocyclic, -SOZ-
substituted heterocyclic or -SO~NRR, where R is hydrogen or alkyl;
(g) substituted aryloxy and substituted heteroaryloxy with the proviso
that at least one substituent on the substituted aryloxy/heteroaryloxy is
other
than halogen, hydroxyl, amino, nitro, trifluoromethyl, trifluoromethoxy,
alkyl, alkenyl, alkynyl, 1,2-dioxymethylene, 1,2-dioxyethylene, alkoxy,
alkenoxy, alkynoxy, alkylamino, alkenylamino, alkynylamino,
alkylcarbonyloxy, acyl, alkylcarbonylamino, alkoxycarbonylamino,
alkylsulfonylamino, N-alkyl or N,N-dialkylurea;
(h) -alkoxy-saturated heterocyclic, -alkoxy-saturated substituted
heterocyclic, -substituted alkoxy-heterocyclic and -substituted alkoxy-
substituted saturated heterocyciic;
(i) -O-heterocyclic and -O-substituted heterocyclic;
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(j) tetrazolyl;
(k) -NR-SO,-substituted alkyl where R is hydrogen, alkyl or aryl,
with the proviso that at least one substituent on the alkyl moiety of the
substituted alkylsulfonylamino is other than halogen, hydroxyl, amino, vitro,
5 trifluoromethyl, trifluoromethoxy, alkyl, alkenyl, alkynyl, 1,2-
dioxymethylene, 1,2-dioxyethylene, alkoxy, alkenoxy, alkynoxy,
alkylamino, alkenylamino, alkynylamino, alkylcarbonyloxy, acyl,
alkylcarbonylamino, alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or
N,N-dialkylurea;
10 (1) alkenylsulfonylamino, alkynylsulfonylamino, substituted
alkenylsulfonylamino and substituted alkynylsulfonylamino;
(m) substituted alkoxy with the proviso that the substitution on the
alkyl moiety of said substituted alkoxy does not include alkoxy-NR"R" as
defined above, unsaturated heterocyclyl, alkyloxy, aryloxy, heteroaryloxy,
15 aryl, heteroaryl or aryl/heteroaryl substituted with halogen, hydroxyl,
amino,
vitro, trifluoromethyl, trifluoromethoxy, alkyl, alkenyl, alkynyl, 1,2-
dioxymethylene, 1,2-diaxyethylene, alkoxy, alkenoxy, alkynoxy,
alkylamino, alkenylamino, alkynylamino, alkylcarbonyloxy, acyl,
alkylcarbonylamino, alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or
N,N-dialkylurea;
(n) amidine and amidine substituted with from 1 to 3 substituents
independently selected from the group consisting of alkyl, substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl
and
heterocyclic;
(o) -C(O)NR"'R"' where each R"' is independently selected from the
group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,
substituted
cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic with the proviso that when one R"' is unsaturated
heterocyclic, aryl, heteroaryl or aryl/heteroaryl substituted with halogen,
hydroxyl, amino, vitro, trifluoromethyl, trifluoromethoxy, alkyl, alkenyl,
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16
alkynyl, 1,2-dioxymethylene, 1,2-dioxyethylene, alkoxy, alkenoxy,
alkynoxy, alkylamino, alkenylamino, alkynylamino, alkylcarbonyloxy, acyl,
alkylcarbonylamino, alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or
N,N-dialkylurea, then the other R"' is alkyl, substituted alkyl (other than
unsaturated heterocyclyl substituted-alkyl), cycloalkyl, substituted
cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
heterocyclic or substituted heterocyclic;
(p) -NRZZC(O)-R'8 where R'8 is selected from the group consisting of
alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,
substituted
aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted
heterocyclic, and R22 is alkyl, substituted alkyl, aryl, substituted aryl,
cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl,
heterocyclic or substituted heterocyclic;
(q) -SOZ-aryl, -S02-substituted aryl, -SO~-heteroaryl, -SO,-substituted
heteroaryl or -SO,-alkyl;
(r) -NR'C(O)NR'9R'9 wherein R' is selected from the group
consisting of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl,
substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic and each R'9 is independently selected from the
group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,
substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic:
(s) -NR'C(O)OR'9 wherein R' is selected from the group consisting
of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl. substituted
cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted
heterocyclic and R'9 is selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic;
(t) -aminocarbonyl-(N-formylheterocylcyl); and
(u) -alkyl-C(O)NH-heterocyclyl and -alkyl-C(O)NH-substituted
heterocyclyl;
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17
and pharmaceutically acceptable salts thereof;
and still further with the following provisos excluding the following
compounds:
A. when R' is p-methylphenyl, Rz and R' together with their pendent
nitrogen and carbon atoms form a pyrrolidinyl ring and Q is -C(O)NH-, then
RS is not -CHZCOOH or -CHZCHZCOOH; and
B. when R' is p-methylphenyl, Rz and R3 together with their pendent
nitrogen and carbon atoms form a pyrrolidinyl ring and Q is -C(O)NH-, then
RS is not 2,4,6-trimethylbenzyl.
In another embodiment, the compounds of this invention can also be
provided as prodrugs which convert (e.g., hydrolyze, metabolize, etc.) in vivo
to a compound of formula I above. In a preferred example of such an
embodiment, the carboxylic acid in the compound of formula I is modified
into a group which, in vivo, will convert to the carboxylic acid (including
salts thereof). In a particularly preferred embodiment, such prodrugs are
represented by compounds of formula IA:
R' O
~ II
R'-SO,-N(R'-)-C-Q-CH-C-R6 IA
H RS
where
R' is selected from the group consisting of alkyl, substituted alkyl,
aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heterocyclic,
substituted heterocylic, heteroaryl and substituted heteroaryl;
R'- is selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,
substituted
cycloalkenyl, heterocyclic, substituted heterocyclic, aryl, substituted aryl,
heteroaryl, and substituted heteroaryl, and R' and R'- together with the
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18
nitrogen atom bound to RZ and the SOZ group bound to R' can form a
heterocyclic or a substituted heterocyclic group;
R3 is selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, heterocyclic, and substituted
heterocyclic,
and Rz and R3 together with the nitrogen atom bound to Rz and the carbon
atom bound to R3 can form a saturated heterocyclic group or a saturated
substituted heterocyclic group with the proviso that when monosubstituted,
the substituent on said saturated substituted heterocyclic group is not
carboxyl;
Ar is aryl, heteroaryl, substituted aryl or substituted heteroaryl;
x is an integer of from 1 to 4;
R6 is selected from the group consisting of 2,4-dioxo-tetrahydrofuran-
3-yl (3,4-enol), amino, alkoxy, substituted alkoxy, cycloalkoxy, substituted
cycloalkoxy, -O-(N-succinimidyl), -NH-adamantyl, -O-cholest-5-en-3-(3-yl,
-NHOY where Y is hydrogen, alkyl, substituted alkyl, aryl, and substituted
aryl, -NH(CHZ)PCOOY where p is an integer of from I to 8 and Y is as
defined above, -OCHZNR9R'° where R9 is selected from the group
consisting
of -C(O)-aryl and -C(O)-substituted aryl and R'° is selected from the
group
consisting of hydrogen and -CH,COOR" where R" is alkyl, and -NHSO,Z"
where Z" is alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl,
aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic or
substituted
heterocyclic;
Q is -C(X)NR'- wherein R' is selected from the group consisting of
hydrogen and alkyl;
X is selected from the group consisting of oxygen and sulfur;
RS is -CH~X where X is selected from the group consisting of
hydrogen, hydroxyl, acylamino, alkyl, alkoxy, aryloxy, aryl, aryloxyaryl,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
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carboxyl-substituted heterocyclic, cycloalkyl, substituted alkyl, substituted
alkoxy, substituted aryl, substituted aryloxy, substituted aryloxyaryl,
substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic;
with the further provisos that:
A. RS is not selected from the group consisting of -(CHZ)n aryl
and -(CHZ)"heteroaryl where n is an integer equal to 1 to 4 when RZ and R3
together with the nitrogen atom bound to R'- and the carbon atom bound to R3
form a saturated heterocyclic group or a saturated substituted heterocyclic
group;
B. RS is not -(CHZ)X Ar-RS' where RS' is selected from the group
consisting of -O-Z-NRBR$' and -O-Z-R'Z wherein Rg and R$' are
independently selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic, and
substituted heterocyclic, and where R$ and Rg' are joined to form a
heterocycle or a substituted heterocycle; R''- is selected from the group
consisting of heterocycles and substituted heterocycles, and Z is selected
from the group consisting of -C(O)- and -SOZ-,
Ar is aryl, heteroaryl, substituted aryl or substituted heteroaryl,
x is an integer of from 1 to 4;
C. RS is not -(CH,)X-Ar-RS' where RS' is selected from the group
consisting of -NR24C(Z')NRgR$' and -NR24C(Z')R'3 wherein Z' is selected
from the group consisting of oxygen, sulfur and NR'-4, R'-4 is selected from
the group consisting of hydrogen, alkyl and aryl, R8 and R8~ are
independently selected from the group consisting of hydrogen, alkyl,
substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,
heterocyclic, substituted heterocyclic, heteroaryl and substituted heteroaryl
provided that when Z' is oxygen, at least one of R8 and R8' is substituted
alkyl, cycloalkyl, substituted cycloalkyl, saturated heterocyclic other than
morpholino and thiomorpholino, and substituted heterocyclic, or R8 and R$'
can be joined to form a saturated heterocycle other than morpholino or
*rB
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thiomorpholino, a saturated substituted heterocycle or a saturated/unsaturated
heterocycle having an amino group substituted with an alkoxycarbonyl
substituent, and further provided that when Z' is sulfur, at least one of R$
and
Rg' is a group other than aryl, substituted aryl, heteroaryl or substituted
5 heteroaryl, R'3 is selected from the group consisting of substituted
heterocyles and saturated heterocycles other than morpholino and
thiomorpholino,
Ar is aryl, substituted aryl, heteroaryl or substituted heteroaryl,
x is an integer of from 1 to 4;
10 D. RS is not -ALK-X where ALK is an alkyl group of from 1 to
10 carbon atoms attached via a methylene group (-CH,-) to the carbon atom
to which it is attached; X is selected from the group consisting of
substituted
alkylcarbonylamino, substituted alkenylcarbonylamino, substituted
alkynylcarbonylamino, heterocyclylcarbonylamino, substituted
15 heterocyclylcarbonylamino, acyl, acyloxy, aminocarbonyloxy, acylamino,
oxycarbonylamino, alkoxycarbonyl, substituted alkoxycarbonyl,
aryloxycarbonyl, substituted aryloxycarbonyl, cycloalkoxycarbonyl,
substituted cycloalkoxycarbonyl, heteroaryloxycarbonyl, substituted
heteroaryloxycarbonyl, heterocyclyloxycarbonyl, substituted
20 heterocyclyloxycarbonyl, cycloalkyl, substituted cycloalkyl, saturated
heterocyclic, substituted saturated heterocyclic, substituted alkoxy,
substituted alkenoxy, substituted alkynoxy, heterocyclyloxy, substituted
heterocycloxy, substituted thioalkyl, substituted thioalkenyl, substituted
thioalkynyl, aminocarbonylamino, aminothiocarbonylamino, guanidino,
amidino, alkylamidino, thioamidino, halogen, cyano, nitro, -OS(O)2-alkyl,
-OS(O)2-substituted alkyl, -OS(O)~-cycloalkyl, -OS(O),-substituted
cycloalkyl, -OS(O)~-aryl, -OS(O)~-substituted aryl, -OS(O)~-heteroaryl,
-OS(O)z-substituted heteroaryl, -OS(O),-heterocyclic, -OS(O)2-substituted
heterocyclic, -OSOz-NRR, -NRS(O)~-alkyl, -NRS(O),-substituted alkyl,
-NRS(O),-cycloalkyl, -NRS(O),-substituted cycloalkyl, -NRS(O)~-aryl,
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21
-NRS(O)2-substituted aryl, -NRS(O),-heteroaryl, -NRS(O)2-substituted
heteroaryl, -NRS(O)2-heterocyclic, -NRS(O),-substituted heterocyclic,
-NRS(O)2-NR-alkyl, -NRS(O)2-NR-substituted alkyl, -NRS{O)2-NR-
cycloalkyl, -NRS(O)2-NR-substituted cycloalkyl, -NRS(O)z-NR-aryl,
-NRS(O)2-NR-substituted aryl, -NRS(O)2-NR-heteroaryl, -NRS(O)2-NR-
substituted heteroaryl, -NRS(O)Z-NR-heterocyclic, -NRS(O)z-NR-substituted
heterocyclic where R is hydrogen or alkyl, -S(O),-alkyl, -S(O)2-substituted
alkyl, -S(O)2-aryl, -S(O)z-substituted aryl, -S(O),-substituted heteroaryl,
-S(O),-substituted heteroaryl, -S(O)2-heterocycIic, -S(O)S-substituted
heterocyclic, mono- and di-(substituted alkyl)amino, N,N-(alkyl, substituted
alkyl)amino, N,N-(aryl, substituted alkyl)amino, N,N-(substituted aryl,
substituted alkyl)amino, N,N-{heteroaryl, substituted alkyl)amino, N,N-
(substituted heteroaryl, substituted alkyl)amino, N,N-(heterocyclic,
substituted alkyl)amino, N,N-N,N-(substituted heterocyclic, substituted
alkyl)amino, mono- and di-(heterocyclic)amino, mono- and di-(substituted
heterocyclic)amino, N,N-(alkyl, heterocyclic)amino, N,N-(alkyl, substituted
heterocyclic)amino, N,N-(aryl, heterocyclic)amino, N,N-(substituted aryl,
heterocyclic)amino, N,N-(aryl, substituted heterocyclic)amino, N,N-
(substituted aryl, substituted heterocyclic)amino, N,N-(heteroaryl,
heterocyclic)amino, N,N-(heteroaryl, substituted heterocyclic)amino, N,N-
(substituted heteroaryl, heterocyclic)amino, and N,N-(substituted heteroaryl,
substituted heterocyclic)amino; and
E. RS is not -{Cl-iz)X Ar-Rsu where RS~~ is a substituent selected
from the group consisting of:
(a) substituted alkylcarbonylamino with the proviso that at least one
of the substituents on the substituted alkyl moiety is selected from the group
consisting of alkoxy, substituted alkoxy, acyl, acylamino,
thiocarbonylamino, acyloxy, alkenyl, amino, amidino, alkyl amidino,
thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryloxy, substituted aryloxy, cyano, nitro, halogen,
hydroxyl. carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-
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cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-
substituted aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,
substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl,
substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,
thioheterocyclic, substituted thioheterocyclic, heterocyclic, substituted
heterocyclic, cycloalkoxy, substituted cycloalkyoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -OS(O),-alkyl, -OS(O)z-
substituted alkyl, -OS(O)~-aryl, -OS(O)Z-substituted aryl, -OS(O),-heteroaryl,
-OS(O)2-substituted heteroaryl, -OS(O)2-heterocyclic, -OS(O)~-substituted
heterocyclic, -OSOZ-NRR, -NRS(O),-alkyl, -NRS(O)2-substituted alkyl,
-NRS(O)Z-aryl, -NRS(O),-substituted aryl, -NRS(O)z-heteroaryl, -NRS(O)2-
substituted heteroaryl, -NRS(O)Z-heterocyclic, -NRS(O)~-substituted
heterocyclic, -NRS(O),-NR-alkyl, -NRS(O),-NR-substituted alkyl, -
NRS(O)2-NR-aryl, -NRS(O)Z-NR-substituted aryl, -NRS(O)2-NR-heteroaryl,
-NRS(O)~-NR-substituted heteroaryl, -NRS(O),-NR-heterocyclic, -NRS(O),-
NR-substituted heterocyclic, mono- and di-alkylamino, mono- and di-
(substituted alkyl)amino, mono- and di-arylamino, mono- and di-(substituted
aryl)amino, mono- and di-heteroarylamino, mono- and di-(substituted
heteroaryl)amino, mono- and di-heterocyclic amino, mono- and di-
(substituted heterocyclic) amino, unsymmetric di-substituted amines having
different substituents selected from the group consisting of alkyl,
substituted
alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic
and substituted heterocyclic, substituted alkyl groups having amino groups
blocked by conventional blocking groups (such as Boc, Cbz, formyl, and the
like) and alkyl/substituted alkyl groups substituted with -SO~-alkyl, -SOZ-
substituted alkyl, -SO~-alkenyl, -SO~-substituted alkenyl, -SOz-cycloalkyl,
-SOZ-substituted cycloalkyl, -SO,-aryl, -SO~-substituted aryl, -SO,-
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23
heteroaryl, -SOz-substituted heteroaryl, -SOZ-heterocyclic, -SOz-substituted
heterocyclic or -SOZNRR, where R is hydrogen or alkyl;
(b) alkoxyaryl substituted on the alkoxy moiety with a substituent
selected from the group consisting of carboxyl and -COORz3 where Rz3 is
alkyl, substituted alkyl, cycloalkyl, aryl, heteroaryl or heterocyclic;
(c) aryl and heteroaryl;
(d) -NR'R' wherein each R' is independently selected from the group
consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl,
substituted heteroaryl, cycloalkyl, substituted cycloalkyl, heterocyclic and
substituted heterocyclic with the proviso that at least one of R' is
substituted
alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic or substituted
heterocyclic and with the further proviso that when R' is substituted alkyl at
least one of the substituents on the substituted alkyl moiety is selected from
the group consisting of alkoxy, substituted alkoxy, acyl, acylamino,
thiocarbonylamino, acyloxy, alkenyl, amino, amidino, alkyl amidino,
thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryloxy, substituted aryloxy, cyano, nitro, halogen,
hydroxyl, carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-
cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-
substituted aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, thiol, thioalkyl,
substituted thioalkyl, thioaryl, substituted thioaryl, thiocycloalkyl,
substituted thiocycloalkyl, thioheteroaryl, substituted thioheteroaryl,
thioheterocyclic, substituted thioheterocyclic, heterocyclic, substituted
heterocyclic, cycloalkoxy, substituted cycloalkyoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -OS(O),-alkyl, -OS(O)~-
substituted alkyl, -OS(O)Z-aryl, -OS(O)~-substituted aryl, -OS(O),-heteroaryl,
-OS(O),-substituted heteroaryl, -OS(O),-heterocyclic, -OS(O),-substituted
heterocyclic, -OSOz-NRR, -NRS(O)~-alkyl, -NRS(O),-substituted alkyl,
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-NRS(O)2-aryl, -NRS(O)Z-substituted aryl, -NRS(O),-heteroaryl, -NRS(O)2-
substituted heteroaryl, -NRS(O),-heterocyclic, -NRS(O)~-substituted
heterocyclic, -NRS(O)2-NR-alkyl, -NRS(O)2-NR-substituted alkyl,
-NRS(O)2-NR-aryl, -NRS(O)2-NR-substituted aryl, -NRS(O)Z-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS{O)Z-NR-heterocyclic,
-NRS(O)2-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
{substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic, substituted alkyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like) and alkyl/substituted alkyl groups substituted with -
SO,-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SO,-substituted alkenyl, -
SOZ-cycloalkyl, -SOZ-substituted cycloalkyl, -SO,-aryl, -SO,-substituted aryl,
-SO~-heteroaryl, -SO,-substituted heteroaryl, -SO,-heterocyclic, -SOz-
substituted heterocyclic or -SO~NRR, where R is hydrogen or alkyl;
(e) -alkoxy-NR"R" wherein each R" is independently selected from
the group consisting of alkyl, substituted alkyl, aryl, substituted aryl,
cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl,
heterocyclic, and substituted heterocyclic with the proviso that when each R"
is substituted alkyl then at least one of the substituents on the substituted
alkyl moiety is selected from the group consisting of alkoxy, substituted
alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy, alkenyl, amino,
amidino, alkyl amidino, thioamidino, aminoacyl, aminocarbonylamino,
aminothiocarbonylamino, aminocarbonyloxy, aryloxy, substituted aryloxy,
cyano, nitro, halogen, hydroxyl, carboxyl, carboxylalkyl, carboxyl-
substituted alkyl, carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl,
carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-
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substituted heteroaryl, carboxylheterocyclic, carboxyl-substituted
heterocyclic, cycloalkyl, substituted cycloalkyl, guanidine, guanidinosulfone,
thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,
thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substituted
5 thioheteroaryl, thioheterocyclic, substituted thioheterocyclic,
heterocyclic,
substituted heterocyclic, cycloalkoxy, substituted cycloalkyoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -OS(O)~-alkyl,
OS(O),-substituted alkyl, -OS(O)2-aryl, -OS(O),-substituted aryl, -OS(O)2-
10 heteroaryl, -OS(O)2-substituted heteroaryl, -OS(O)~-heterocyclic, -OS(O),-
substituted heterocyclic, -OSOZ-NRR, -NRS(O)~-alkyl, -NRS(O)~-substituted
alkyl, -NRS(O)Z-aryl, -NRS(O)~-substituted aryl, -NRS(O),-heteroaryl,
NRS(O)2-substituted heteroaryl, -NRS(O),-heterocyclic, -NRS(O)2-
substituted heterocyclic, -NRS(O)2-NR-alkyl, -NRS{O)~-NR-substituted
15 alkyl, -NRS(O)2-NR-aryl, -NRS(O),-NR-substituted aryl, -NRS(O),-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O)~-NR-heterocyclic,
-NRS(O)z-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
{substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
20 (substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, and unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic, substituted alkyl groups having
25 amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like) and alkyl/substituted alkyl groups substituted with -
SO~-alkyl, -SOZ-substituted alkyl, -SO~-alkenyl, -SO,-substituted alkenyl, -
SOZ-cycloalkyl, -SO~-substituted cycloalkyl, -SO,-aryl, -SO,-substituted aryl,
-SO,-heteroaryl, -SO,-substituted heteroaryl, -SO~-heterocyclic, -SO,-
substituted heterocyclic or -SO,NRR, where R is hydrogen or alkyl;
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26
(f) substituted alkenyl or substituted alkynyl with the proviso that at
least one of the substituents on the substituted alkenyl/alkynyl moiety is
selected from the group consisting of alkyl, substituted alkyl, aryl,
substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted
heteroaryl, heterocyclic, and substituted heterocyclic with the proviso that
when substituted with substituted alkyl then at least one of the substituents
on the substituted alkyl moiety is selected from the group consisting of
alkoxy, substituted alkoxy, acyl, acylamino, thiocarbonylamino, acyloxy,
alkenyl, amino, amidino, alkyl amidino, thioamidino, aminoacyl,
aminocarbonylamino, aminothiocarbonylamino, aminocarbonyloxy, aryloxy,
substituted aryloxy, cyano, nitro, halogen, hydroxyl, carboxyl, carboxylalkyl,
carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-substituted
cycloalkyl, carboxylaryl, carboxyl-substituted aryl, carboxylheteroaryl,
carboxyl-substituted heteroaryl, carboxylheterocyclic, carboxyl-substituted
heterocycIic, cycloalkyl, substituted cycloalkyl, guanidino, guanidinosulfone,
thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted thioaryl,
thiocycloalkyl, substituted thiocycloalkyl, thioheteroaryl, substituted
thioheteroaryl, thioheterocyclic, substituted thioheterocyclic, heterocyclic,
substituted heterocyclic, cycloalkoxy, substituted cycloalkyoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino, -OS(O)~-alkyl, -
OS(O)~-substituted alkyl, -OS(O),-aryl, -OS(O),-substituted aryl, -OS(O)~-
heteroaryl, -OS(O)~-substituted heteroaryl, -OS(O),-heterocyclic, -OS(O)2-
substituted heterocyclic, -OSO~-NRR, -NRS(O),-alkyl, -NRS(O)2-substituted
alkyl, -NRS(O)2-aryl, -NRS(O)Z-substituted aryl, -NRS(O),-heteroaryl, -
NRS(O),-substituted heteroaryl, -NRS(O)~-heterocyclic, -NRS(O),-
substituted heterocyclic, -NRS(O),-NR-alkyl, -NRS(O),-NR-substituted
alkyl, -NRS(O)2-NR-aryl, -NRS(O)2-NR-substituted aryl, -NRS(O)2-NR-
heteroaryl, -NRS(O)~-NR-substituted heteroaryl, -NRS(O),-NR-heterocyclic,
-NRS(O),-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di
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(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic, substituted alkyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like) and alkyl/substituted alkyl groups substituted with -
SO~-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SOZ-substituted alkenyl, -
SOz-cycloalkyl, -SOZ-substituted cycloalkyl, -SO~-aryl, -SOZ-substituted aryl,
-SO,-heteroaryl, -SOZ-substituted heteroaryl, -SO,-heterocyclic, -SOZ-
substituted heterocyclic or -SO~NRR, where R is hydrogen or alkyl;
(g) substituted aryloxy and substituted heteroaryloxy with the proviso
that at least one substituent on the substituted aryloxy/heteroaryloxy is
other
than halogen, hydroxyl, amino, vitro, trifluoromethyl, trifluoromethoxy,
alkyl, alkenyl, alkynyl, 1,2-dioxymethylene, 1,2-dioxyethylene, alkoxy,
alkenoxy, alkynoxy, alkylamino, alkenylamino, alkynylamino,
alkylcarbonyloxy, acyl, alkylcarbonylamino, alkoxycarbonylamino,
alkylsulfonylamino, N-alkyl or N,N-dialkylurea;
(h) -alkoxy-saturated heterocyclic, -alkoxy-saturated substituted
heterocyclic, -substituted alkoxy-heterocyclic and -substituted alkoxy-
substituted saturated heterocyclic;
(i) -O-heterocyclic and -O-substituted heterocyclic;
(j) tetrazolyl;
{k) -NR-SO~-substituted alkyl where R is hydrogen, alkyl or aryl,
with the proviso that at least one substituent on the alkyl moiety of the
substituted alkylsulfonylamino is other than halogen, hydroxyl, amino, vitro,
trifluoromethyl, trifluoromethoxy, alkyl, alkenyl, alkynyl, 1,2-
dioxymethylene, 1,2-dioxyethylene, alkoxy, alkenoxy, alkynoxy,
alkylamino, alkenylamino, alkynylamino, alkylcarbonyloxy, acyl,
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alkylcarbonylamino, alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or
N,N-dialkylurea;
(1) alkenylsulfonylamino, alkynylsulfonylamino, substituted
alkenylsulfonylamino and substituted alkynylsulfonylamino;
S (m) substituted alkoxy with the proviso that the substitution on the
alkyl moiety of said substituted alkoxy does not include alkoxy-NR"R" as
defined above, unsaturated heterocyclic, alkyloxy, aryloxy, heteroaryloxy,
aryl, heteroaryl or aryl/heteroaryl substituted with halogen, hydroxyl, amino,
nitro, trifluoromethyl, trifluoromethoxy, alkyl, alkenyl, alkynyl, 1,2-
dioxymethylene, 1,2-dioxyethylene, alkoxy, alkenoxy, alkynoxy,
alkylamino, alkenylamino, alkynylamino, aikylcarbonyloxy, acyl,
alkylcarbonylamino, alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or
N,N-dialkylurea;
(n) amidine and amidine substituted with from I to 3 substituents
independently selected from the group consisting of alkyl, substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, heteroaryl
and
heterocyclic;
(o) -C(O)NR"'R"' where each R"' is independently selected from the
group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,
substituted
cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic with the proviso that when one R"' is unsaturated
heterocyclic, aryl, heteroaryl or aryl/heteroaryl substituted with halogen,
hydroxyl, amino, nitro, trifluoromethyl, trifluoromethoxy, alkyl, alkenyl,
alkynyl, 1,2-dioxymethylene, 1,2-dioxyethylene, alkoxy, alkenoxy,
alkynoxy, alkylamino, alkenylamino, alkynylamino, alkyicarbonyloxy, acyl,
alkylcarbonylamino, alkoxycarbonylamino, alkylsulfonylamino, N-alkyl or
N,N-dialkylurea, then the other R"' is alkyl, substituted alkyl (other than
unsaturated heterocyclyl substituted-alkyl), cycloalkyl, substituted
cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
heterocyclic or substituted heterocyclic;
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(p) -NRZZC(O)-R'g where R'$ is selected from the group consisting of
alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl,
substituted
aryl, heteroaryI, substituted heteroaryl, heterocyclic and substituted
heterocyclic, and Rzz is alkyl, substituted alkyl, aryl, substituted aryl,
cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl,
heterocyclic or substituted heterocyclic;
(q) -SOZ-aryl, -SOZ-substituted aryl, -SO,-heteroaryl, -SOZ-substituted
heteroaryl or -SOZ-alkyl;
(r) -NR'C(O)NR'9R'9 wherein R' is selected from the group
consisting of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl,
substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic and each R'9 is independently selected from the
group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl,
substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic:
(s) -NR'C(O)OR'9 wherein R' is selected from the group consisting
of alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted
cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted
heterocyclic and R'9 is selected from the group consisting of hydrogen, alkyl,
substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl,
heteroaryl, substituted heteroaryl, heterocyclic and substituted.
heterocyclic;
(t) -aminocarbonyl-(N-formylheterocylcyl); and
(u) -alkyl-C(O)NH-heterocyclyl and -alkyl-C(O)NH-substituted
heterocyclyl;
and pharmaceutically acceptable salts thereof;
with the following additional proviso:
when R' is p-methylphenyl, RZ and R3 together with their pendent
nitrogen and carbon atoms form a pyrrolidinyl ring. RS is p-[-OCHZCHZ-(4,5-
dihydroimidizol-2-y1), Q is -C(O)NH-, then R6 is not -O-methyl.
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Preferred compounds within the scope of formulas 1 and IA above
include by way of example the following:
N (Toluene-4-sulfonyl)-L-prolyi-4-(a-methyIbenzyloxy)-L-phenylalanine
5 N (Toluene-4-sulfonyl)-L-prolyl-L-tyrosine
N {Toluene-4-sulfonyl)-L-prolyl-4-carboxyphenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-3-(carboxy)phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-4-(2-carboxyphenoxy)-L-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-O-(benzyl)-L-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-4-(iodo)-L-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(methoxy)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-4-nitro-L-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-4-(O-tert-butyl)-L-tyrosine
N {Toluene-4-sulfonyl)-L-prolyl-L-(3,5-diiodo)-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(aminobenzoyl)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-(3-iodo-4-hydroxy)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-chloro)phenylalanine
N-(Toluene-4-sulfonyl)-L-prolyl-L-leucine
N (Toluene-4-sulfonyl)-L-prolyl-L-alanine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(acetamido)-phenylalanine
N-(Toluene-4-sulfonyl)-L-prolyI-L-isoleucine
N-(Toluene-4-sulfonyl)-L-prolyl-L-aspartic acid
N (Toluene-4-sulfonyl)-L-prolyl-L-lysine
N (Toluene-4-sulfonyl)-L-prolyl-L-glutamic acid
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N (Toluene-4-sulfonyl)-L-prolyl-L-(4-dibenzylamino)-phenylalanine methyl
ester
N-(Toluene-4-sulfonyl)-L-prolyl-L-(N benzyl)-histidine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-dibenzylamino)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-methionine
N (Toluene-4-sulfonyl)-L-prolyl-L-serine
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolylL-(N benzyl)-histidine
N (Toluene-4-sulfonyl)-L-prolyl-L-(1-methyl)histidine
N (Toluene-4-sulfonyl)-L-prolyl-D-(N benzyl}histidine
N (Toluene-4-sulfonyl)-L-glutamyl-L-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-L-(N 3-methyl)histidine
N (Toluene-4-sulfonyl)-L-prolyl-a-amino-2,3-dihydro-(1,4-benzodioxin)-6-
propanoic acid
N-(Toluene-4-sulfonyl)-L-prolyl-a-amino-1,3-benzodioxole-5-propanoic
acid
N (Toluene-4-sulfonyl)-L-prolyl-L-valine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-iodo)-phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(aminobenzoyl)phenylalanine methyl
ester
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-chloro)phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-amino)phenylaianine methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-acetamido)phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-(N-benzyl)-histidine
methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-4-(3-3'-tolylureido)-L-phenylalanine
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N-(Toluene-4-sulfonyl-L-prolyl-4-[(2,3,3a,7a-tetrahydro-I H-indole-2-
carbonyl)-amino]-L-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-pentylamino)phenylalanine methyl
ester
N (Toluene-4-sulfonyl)-L-prolyl-L-O-(2-dibenzylamino-ethyl)-tyrosine
methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-O-[2-(dibenzylamino)ethylj-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-pentylamino)phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(4-chlorobenzylamino)-phenylalanine
methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[3-(4-cyanophenyl)-ureido]-phenyl-
alanine methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[3-(4-cyanophenyl)-ureido]-phenyl-
alanine
30
N (Toluene-4-sulfonyl)-L-prolyl-L-O-(tent-butoxycarbonylmethyl)-tyrosine
methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-O-(tent-butoxycarbonylmethyl)-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[(3S)-3,4-dihydro-isoquinolin-3-yl-
aminocarbonyl]-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-4-[3-(3-methoxy-phenyl)-ureido]-L-
phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-4-[3-(3-methoxy-phenyl)-ureido]-L-
phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-O-(4,5-dihydro-1 H-imidazol-2-yl-
methyl)-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(3-propyl-ureido)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-benzylamino)phenylalanine
IV=(Toluene-4-sulfonyl)-L-prolyl-L-(4-benzylamino)phenylalanine methyl
ester
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S
N (Toluene-4-sulfonyl)-L-prolyl-L-4-{4-chlorobenzylamino)-phenylalanine
N {Toluene-4-sulfonyl)-L-prolyl)-L-(4-chloromethanesulfonylamino)-
phenylalanine
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-4-
(aminobenzoyl)phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-(S,S-dirnethyl)-thiaprolyl-L-4-
(benzamido)phenylalanine
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-tyrosine ethyl ester
N (Toluene-4-Sulfonyl)-L-(S,S-dimethyl}-thiaprolyl-L-tyrosine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[(pyridin-4-
yl)methylamino]phenylanine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[(pyridin-4-
yl)methylamino]phenylanine methyl ester
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-4-(pyridin-3-
carboxamido)phenylalanine methyl ester
2S N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-4-(pyridine-3-
carboxamido)phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[(pyridin-3-
ylmethyl)amino]phenylalanine
3S
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-4-nitrophenylalanine
methyl ester
N (Toluene-4-suIfonyl)-L-prolyl-L-4-nitrophenylalanine ethyl ester
N (Toluene-4-sulfonyl}-L-prolyl-L-4-(2-methoxybenzamido)phenylalanine
ethyl ester
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-(4-nitro)phenylalanine
ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(2-bromobenzamido)phenylalanine
ethyl ester
4S N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiamorphyl-L-4-
aminophenylalanine ethyl ester
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N (Toluene-4-sulfonyl)-L-prolyl-L-4-acetamidophenylalanine ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(2-methoxybenzamido)phenylalanine
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L-4-
acetamidophenylalanine ethyl ester
1S
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L-4-
acetamidophenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-4-acetamidophenylalanine isopropyl ester
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-L-4-(isonicotinamido)
phenylalanine ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(isonicotinamido)phenylalanine
ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-(~t-toluene-4-sulfonyl)histidine methyl
ester
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L-4-
(nicotinamido)phenylalanine ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-(O-methyl)tyrosine ethyl ester
N (a-Toluenesulfonyl)-L-prolyl-L-4-(isonicotinamido)phenylalanine ethyl
ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(2-bromobenzamido)phenylalanine
N (a-Toluenesulfonyl)-L-prolyl-L-4-(isonicotinamido)phenylalanine
N (Toluene-4-sulfonyl)-L-(1,1-dioxo)thiamorpholyl-L-4-(2-
bromobenzamido))phenylalanine ethyl ester
N (~'oluene-4-sulfonyl)-L-(1,1-dioxo)thiamorpholyl-L-4-(2-
bromobenzamido)phenylalanine
N-(Toluene-4-sulfonyl)-L-(5,5-dimethyl)-L-4-
(isonicotinamido)phenylalanine
N (a-Toluenesulfonyl)-L-prolyl-L-4-(2-bromobenzamido)phenylalanine
ethyl ester
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3S
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-tyrosine isopropyl
ester
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-tyrosine tent-butyl
S ester
N (Toluene-4-sulfonyl)-L-prolyl-L-tyrosine tert-butyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(2-
trifluoromethylbenzamido)phenylalanine ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(2-methylbenzamido)phenylalanine
ethyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-4-{2-trifluoromethylbenzamido)
phenylalanine
N (4-Fluorobenzenesulfonyl)-L-thiaprolyl-L-tyrosine tert-butyl Ester
N (4-Fluorobenzenesulfonyl)-L-(S,S-dimethyl)-thiaprolyl-L-tyrosine tert-
butyl ester
N-{Toluene-4-sulfonyl)-L-prolyl-4-(dimethylamino)-L-phenylalanine
2S N-(Toluene-4-sulfonyl)-L-prolyl-4-[(2-bromo)benzamidoJ-L-phenylalanine
ethyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-[(pyrazin-2-yl)C(O)NH)-L-phenylalanine
methyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(4-nitrobenzoyl)-L-phenylalanine ethyl
ester
N-(Toluene-4-sulfonyl)-L-(S,S-dimethyl)thiaprolyl-4-(2-bromobenzarnido)-
3S L-phenylalanine t-butyl ester
N-(Toluene-4-sulfonyl)-L-(S,S-dimethyl)thiaprolyl-4-(2-bromobenzamido)-
L-phenylalanine t-butyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(1-H-2-oxo-3-
methyltetrahydropyrimidin-1-yl)-L-phenylalanine t-butyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-( 1-H-2-oxo-3-
methyltetrahydropyrimidin-1-yl)-L-phenylalanine
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N-(Toluene-4-sulfonyl)-L-prolyl-3-chloro-4-(t-butoxy)-L-phenylalanine t-
butyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-3-chloro-4-(hydroxy)-L-phenylalanine t-
butyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(N,N-dimethylureido)-L-phenylalanine t-
butyl ester
N-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyi-3-chloro-4-
(hydroxy)-L-phenylalanine t-butyl ester
N-(4-fluorobenzenesulfonyl)-L-{5,S-dimethyl)thiaprolyl-3-chloro-4-
(hydroxy)-L-phenylalanine isopropyl ester
20
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methoxyphenyl)-L-phenylalanine t-
butyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methoxyphenyl)-L-phenylalanine
N-(Toluene-4-sulfonyl)-L-prolyl-[(5-N,N-dimethyl ureido)-pyridin-2-yl]-
alanine
N-(Toluene-4-sulfonyl)-L-prolyl-4-(N,N-dimethylsulfamyl)-L-phenylalanine
N-(Toluene-4-sulfonyl)-L-prolyl-4-(N',N',N'--trimethylsulfamyl)-L-
phenylalanine
N-(Toluene-4-sulfonyl)-L-prolyl-( 1-t-butoxycarbonylmethylimidazol-4-yl)-
L-alanine methyl ester
N-{Toluene-4-sulfonyl)-L-prolyl-[N,N-
dimethylaminocarbonylmethylimidazol-4-yl]-L-alanine methyl ester
N-[4-(dimethyl ureylenyl)-benzenesulfonyl]-L-prolyl-4-hydroxy-L-
phenylalanine isopropyl ester
N-(Toluene-4-sulfonyl)-L-(S,5-dimethyl)thiaprolyl-3-chloro-4-hydroxy-L-
phenylalanine isopropyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-3-chloro-4-hydroxy-L-phenylalanine
isopropyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(carboxymethyl)-L-phenylalanine methyl
ester
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N-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methylbenzamido)-L-phenylalanine
methyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(N,N-dimethylaminocarbonylmethyl)-L-
phenylalanine
N-( 1-methylimidazol-4-sulfonyl)-L-prolyl-4-hydroxy-L-phenylalanine
isopropyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2,4,5-trioxo-3-
phenyltetrahydroimidazol-1-yl)-L-phenylalanine isopropyl ester
N-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-3-fluoro-4-
hydroxy-L-phenylalanine isopropyl ester
N-(4-Fluorobenzenesulfonyl)-L-(5,S-dimethyl)thiaprolyl-3-chloro-4-t-
butoxy-L-phenylalanine t-butyl ester
N-{N-[(1S)-2,10-camphorsultamyl~acetyl}-L-tyrosine t-butyl ester
N-{N-[(1S)-2,10-camphorsultamyl]acetyl}-3-chlorotyrosine isopropyl ester
N-(4-Fluorobenzenesulfonyl)-L-thiaprolyl-4-hydroxy-L-phenylalanine t-
butyl ester
N-(Toluene-4-sulfonyl)-L-prolyl-4-(N,N-dimethylaminocarbonylmethyl)-L-
phenylalanine t-butyl ester
N-(4-Nitrobenzenesulfonyl)-L-prolyl-4-(hydroxy)-L-phenylalanine t-butyl
ester
N-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)thiaprolyl-4-hydroxy-L-
phenylalanine isopropyl ester
N-(4-Fluorobenzenesulfonyl)-L-prolyl-4-hydroxy-L-phenylalanine t-butyl
ester
N-(pyridin-3-sulfonyl)-L-prolyl-4-hydroxy-L-phenylalanine t-butyl ester
N-{Toluene-4-sulfonyl)-L-prolyl-4-(methanesulfonamido)-L-phenylalanine
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2,4, 5-trioxo-3-(3-chlorophenyl)-
tetrahydroimidazol-1-yl)-L-phenylalanine benzyl ester
N-(1-methylimidazol-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-3-chloro-4-
hydroxy-L-phenylalanine ethyl ester.
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N-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl) thiaprolyl-L-3-chloro-
4-~-butoxy-phenylalanine ~-Butyl Ester
N-[2-(N-2,10-camphorsultamyl) acetyl]-L-3-chloro-4-hydroxyphenylalanine
Isopropyl Ester
Preferably, in the compounds of formula I and IA above, R' is
selected from the group consisting of alkyl, substituted alkyl, aryl,
substituted aryl, heterocyclic, substituted heterocylic, heteroaryl and
substituted heteroaryl. Even more preferably, R' is selected from the group
consisting of 4-methylphenyl, methyl, benzyl, n-butyl, 4-chlorophenyl, 1-
naphthyl, 2-naphthyl, 4-methoxyphenyl, phenyl, 2,4,6-trimethylphenyl,
2-(methoxycarbonyl)phenyl, 2-carboxyphenyl, 3,5-dichlorophenyl,
4-trifluoromethylphenyl, 3,4-dichlorophenyl, 3,4-dimethoxyphenyl,
4-(CH3C(O)NH-)phenyl, 4-trifluoromethoxyphenyl, 4-cyanophenyl,
isopropyl, 3,5-di-(trifluoromethyl)phenyl, 4-t-butylphenyl, 4-t-butoxyphenyl,
4-nitrophenyl, 2-thienyl, 1-N-methyl-3-methyl-5-chloropyrazol-4-yi,
phenethyl, 1-N-methylimidazol-4-yl, 4-bromophenyl, 4-amidinophenyl,
4-methylamidinophenyl, 4-[CH3SC(=NH)]phenyl, 5-chloro-2-thienyl, 2,5-
dichloro-4-thienyl, 1-N-methyl-4-pyrazolyl, 2-thiazolyl, 5-methyl-1,3,4-
thiadiazol-2-yl, 4-[HZNC(S)]phenyl, 4-aminophenyl, 4-fluorophenyl,
2-fluorophenyl, 3-fluorophenyl, 3,5-difluorophenyl, pyridin-3-yl, pyrimidin-
2-yl, 4-(3'-dimethylamino-n-propoxy)-phenyl, and 1-methylpyrazol-4-yl.
Preferably, in the compounds of formula I and IA above, RZ is
hydrogen, methyl, phenyl, benzyl, -(CHZ),-2-thienyl, and -(CH~)z-~.
In one embodiment, R' and RZ together with the nitrogen atom bound
to RZ and the SO, group bound to R' are joined to form a heterocyclic group
or substituted heterocyclic group. Preferred heterocyclic and substituted
heterocyclic groups include those having from 5 to 7 ring atoms and having 2
to 3 heteroatoms in the ring selected from the group consisting of nitrogen,
oxygen and sulfur, which ring is optionally fused to another ring such as a
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
39
phenyl or cyclohexyl ring to provide for a fused ring heterocycle of from 10
to 14 ring atoms and having 2 to 4 heteroatoms in the ring selected from the
group consisting of nitrogen, oxygen and sulfur. Specifically preferred R'/RZ
joined groups include, by way of example, benzisothiazoIinyl (saccharin-2-
yl).
Preferably, in the compounds of formula I and IA above, R3 includes
all of the isomers arising by substitution with methyl, phenyl, benzyl,
diphenylmethyl, -CH,CHZ-COOH, -CHZ-COOH, 2-amidoethyl, iso-butyl,
t-butyl, -CHZO-benzyl and hydroxymethyl.
In another preferred embodiment, R'- and R3 together with the
nitrogen atom bound to Rz and the carbon atom bound to R' form a saturated
heterocyclic group or a saturated substituted heterocyclic group with the
proviso that when monosubstituted, the substituent on said saturated
substituted heterocyclic group is not carboxyl.
Q is preferably -C(O)NH- or -C(S)NH-.
RS is preferably selected from the group consisting of all possible
isomers arising by substitution with the following groups: p-[-OCH(CH3)~)-
benzyl, 4-hydroxybenzyl, 2-carboxybenzyl, 3-carboxybenzyl,
4-carboxybenzyl, 4-(2-carboxyphenoxy)benzyl, 4-(benzyloxy)benzyl,
4-iodobenzyl, 4-methoxybenzyl, 4-nitrobenzyl, 4-(tert-butoxy)benzyl, 3,5-
diiodo-4-hydroxybenzyl, 4-(benzamido)benzyl, benzyl, 4-hydroxy-3-
iodobenzyl, 4-chlorobenzyl, isobutyl, methyl, 4-(acetamido)benzyl, n-butyl,
carboxymethyl, 4-aminobutyl, 2-carboxyethyl, 4-(N,N dibenzylamino)-
benzyl, (N benzylimidazol-4-yl)methyl, 2-thiomethoxyethyl, hydroxymethyl,
(N methylimidazol-4-yl)methyl, 4-(isopropyl-C(O)NH-) butyl,
4-{benzarnido)butyl, 4-(benzyl-C(O)NH-)butyl, (N methylimidazol-5-
yl)methyl, 4-(pyridin-2-yl-C(O)NH-)butyl, 4-(6-methylpyridin-3-yl-
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
C(O)NH-) butyl, 4-(3-methylthien-2-yl-C(O)NH-)butyl, 4-(pyrrol-2-yl-
C(O)NH-)butyl, 4-(furan-2-yl-C(O)NH)-butyl, isopropyl, 4-aminobenzyl, 4-
(4-phenylbutoxy)benzyl, 4-(1-methylindol-3-yl-C(O)NH-)butyl, 4-(4-
methanesulfonylphenyl-C(O)NH-)butyl, 4-(4-acetylphenyl-C(O)NH-)butyl,
5 4-(4-fluorophenyl-C(O)NH-)butyl, 4-[2-(pyridin-2-yl)ethynylJbenzyl,
4-[2-(3-hydroxyphenyl)ethynyl]benzyl, 4-(pyridin-4-yl-C(O)NH-)benzyl,
4-(pyridin-3-yl-C(O)NH-)benzyl, 4-(3-methylphenyl-NHC(O)NH-)benzyl,
4-(2,3-dihydroindol-2-yl-C(O)NH-)benzyl, 4-(N,N dipentylarnino)benzyl,
4-(N pentylamino)benzyl, 4-[2-(N,N dibenzylamino)ethoxy]benzyl,
10 3-hydroxybenzyl, 4-(N n-butyl-N n-pentylamino)benzyl, 4-(N 4-
chlorophenylamino)benzyl, 4-(4-cyanophenyl-NHC(O)NH-)benzyl,
4-(carboxymethoxy)benzyl, 4-(tert-butoxycarbonylmethoxy)benzyl,
4-(5-fluoroindol-2-yl-C(O)NH-)benzyl, 4-(1,2,3,4-tetrahydroisoquinolin-3-
yl-C(O)NH-)benzyl, 4-(3-methoxyphenyl-NHC(O)NH-)benzyl, 4-[2-(indol-
15 3-yl)ethoxy]benzyl, 4-(4,S-dihydroimidazol-2-ylmethoxy)benzyl, 4-(n-
propyl-NHC(O)NH-)benzyl, 4-(N benzylamino)benzyl, 3-methoxybenzyl, 4-
(pyridin-2-yl-C(O)NH-)benzyl, 4-(N 4-chlorobenzylamino)benzyl, 4-(2-
chloromethylsulfonylamino)benzyl, 4-(N,N-dimethylamino)benzyl,
3-aminobenzyl, 4-(benzyl)benzyl, 2-hydroxyethyl, 4-nitrobenzyl, 4-(phenyl-
20 NHC(S)NH-)benzyl, 4-(pyridin-3-yl-NHC(S)NH-)benzyl, 4-(pyridin-4-
ylmethylamino)benzyl, 4-[~CHzOCHz(Boc-HN)CHC(O)NH-]benzyl,
4-(pyridin-3-yl-C(O)NH-)butyl, 4-(pyridin-4-yl-C(O)NH-)butyl, 4-(pyridin-
3-yl-C(O)NH-)benzyl, 4-(pyridin-4-yl-C(O)NH-)benzyl, 4-(N
toluenesulfonylpyrrolidin-2-yl-C(O)NH-)butyl, 4-(piperidin-4-yl-C(O)NH-)
25 butyl, 4-(2-Boc-1,2,3,4-tetrahydroisoquinolin-3-yl-NHCH,-)benzyl, 4-(2-
Boc-1,2,3,4-tetrahydroisoquinolin-3-yl-C(O)NH-)benzyl, 4-{pyridin-3-
ylmethylamino)benzyl, 4-[c~CH20(O)C(CbzNH)CHCH,CH,C(O)NH-]
benzyl, 4-(2-methoxybenzamido)benzyl, 4-(2-bromobenzamido)benzyl,
4-(pyrazin-2-yl-C(O)NH-)benzyl, (1-toluenesulfonylimidizol-4-yl)methyl,
30 [ 1-(N,N-dimethylaminosulfonyl)imidizol-4-yl]methyl, 4-(trifluoromethyl)-
benzyl, 4-(3,3-dimethylureido)benzyl, 4-(methoxycarbonylamino)benzyl, 4-
CA 02290748 1999-11-19
WO 99/Ob437 PCT/US98/16070
41
(1,3,3-trimethylureido)benzyl, 4-(methoxycarbonyl-N methylamino)benzyl,
4-cyanobenzyl, 4-(2-formyl-1,2,3,4-tetrahydroisoquinolin-3-yl-
C(O)NH)benzyl, phenyl, 4-(aminomethyl)benzyl, 4-(1-Boc-piperidin-4-yl-
C(O)NHCHZ-)benzyl, 4-(1-Boc-piperidin-4-yl-C(O)O-)benzyl, 4-(piperidin-
4-yl-C(O)NHCH2-)benzyl, 4-[(1-methylpiperidin-4-yl)-O-]benzyl, 4-
(1,2,3,4-tetrahydroquinolin-2-yl-C(O)NH-)benzyl, a-methylbenzyl,
4-(trimethylacetamido)benzyl, 4-(2-methylpropionamido)benzyl, 4-
(morpholin-4-yl-C(O)NH-)benzyl, 4-(3,3-diethylureido)benzyl, 4-(2-
trifluoromethylbenzamido)benzyl, 4-(2-methylbenzamido)benzyl, 4-
hydroxy-3-nitrobenzyl, 3-hydroxy-4-(~-OC(O)NH-)benzyl, 4-
(thiomorpholino-4-yl-C(O)NH-)benzyl, 4-(l,l-dioxothiomorpholino-4-yl-
C(O)NH-)benzyl, 3-nitro-4-(methoxycarbonylmethoxy)benzyl, (2-
benzoxazoIinon-6-yl)methyl, (2H-1,4-benzoxazin-3(4H)-one-7-yl)methyl, 4-
[N,N dimethyaminosulfonyl-(N methyl)amino]benzyl, 4-[(2-
methylpyrrolidin-1-yl)-C(O)NH-]benzyl, (pyridin-4-yl)methyl, 4-(1-
methylpiperidin-4-yl-C(O)NH-)benzyl, 4-[bis(N,N-
dimethylaminothiocarbonyl)amino]benzyl, 4-(N,N-dimethylaminosulfonyl)-
benzyl, 4-(imidazolid-2-one-1-yl)benzyl, 3,4-(ethylenedioxy)benzyl-, 3,4-
(methylenedioxy)benzyl- and 4-(3-formylimidazolid-2-one-1-yl)benzyl.
In the compounds of formula IA, R6 is preferably 2,4-dioxo-
tetrahydrofuran-3-yl (3,4-enol), methoxy, ethoxy, iso-propoxy, n-butoxy,
t-butoxy, cyclopentoxy, neo-pentoxy, 2-a-iso-propyl-4-[i-
methylcyclohexoxy, 2-[3-isopropyl-4-[3-methylcyclohexoxy, -NHz,
benzyloxy, -NHCHZCOOH, -NHCH,CH,COOH, -NH-adamantyl, -
NHCH,CHZCOOCHzCH3, -NHSOz p-CH3-~, -NHORg where R8 is
hydrogen, methyl, iso-propyl or benzyl, O-(N-succinimidyl), -O-cholest-5-
en-3-(3-yh -OCH,-OC(O)C(CH3)3, -O(CH~)ZNHC(O)W where z is 1 or 2 and
W is selected from the group consisting of pyrid-3-yl, N-methylpyridyl, and
N-methyl-1,4-dihydro-pyrid-3-yl, -NR"C(O)-R' where R' is aryl, heteroaryl
or heterocyclic and R" is hydrogen or -CH,C(O)OCH,CH3.
CA 02290748 1999-11-19
WO 99/06437 PCTlUS98/16070
42
This invention also provides methods for binding VLA-4 in a
biological sample which method comprises contacting the biological sample
with a compound of formula I or IA above under conditions wherein said
compound binds to VLA-4.
Certain of the compounds of formula I and IA above are also useful
in reducing VLA-4 mediated inflammation in vivo.
This invention also provides pharmaceutical compositions
comprising a pharmaceutically acceptable carrier and a therapeutically
effective amount of one or more of the compounds of formula I or IA above
with the exception that R3 and RS are derived from L-amino acids or other
similarly configured starting materials. Alternatively, racemic mixtures can
be used.
The pharmaceutical compositions may be used to treat VLA-4
mediated disease conditions. Such disease conditions include, by way of
example, asthma, Alzheimer's disease, atherosclerosis, AIDS dementia,
diabetes (including acute juvenile onset diabetis), inflammatory bowel
disease (including ulcerative colitis and Crohn's disease), multiple
sclerosis,
rheumatoid arthritis, tissue transplantation, tumor metastasis, meningitis,
encephalitis, stroke, and other cerebral traumas, nephritis, retinitis, atopic
dermatitis, psoriasis, myocardial ischemia and acute leukocyte-mediated lung
injury such as that which occurs in adult respiratory distress syndrome.
Accordingly, this invention also provides methods for the treatment
of an inflammatory disease in a patient mediated by VLA-4 which methods
comprise administering to the patient the pharmaceutical compositions
described above.
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
43
Preferred compounds of formula I and IA above include those set
forth in Tables IA and IB below.
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
44
x
z
U
x x x x x
,
II
a
oc o 0 o z
, , Q Q
, _~ _,
N
C
~ n
2 K
U E
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s
~
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r3
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a a
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a
I n H U H U H V U H
.. ~ ~. n H ~
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E>,
~, ;, ;,
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=_ =_ = i
re Wo ;g
v
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Z 0.! II II II _ II
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~~ n~
, x a. . o
x " a "
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. 7
1
~ ~ ~ C~ t~-,~
~ ,
ac
,
U U U U U
b
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
."
s x x x x x x x
a
a
~c o 0 0 0 0 0 0 0
s
N
C V
T j,
~ ~ C
N n
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p
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U
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z_
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d
~ ~ ~~, Vf N ~ V1 H
U H U H U H U H .~~ U ~ U V ..~~ U ~
E >, ~ E r>. ~ E >, ~ E a, ~ E >, ~ E i, ~ E ~~ ' E a,
~ U C V C U C_ U C_ U p C_ U _Q C_ V _Q C . U _p C_
U ~ b U ~ ;b U ~ ,Q U ~ ,0 V ~ b U ~ ,~ U ~ ,p V ~ ,p
pG II ° ~ II ° ~ II ° ~ II C ~ II C ~ II ~ ~ ~I ~ II
n n n
x a x .~ x Q oc J x a x .a oc ~ ,.~ x
f"1 ~ G~" t'~Y 0~, (n5 v ~ f'~1 ar tr1 v C14 c'1 yr ~" t'~S v
-fi -s. -6w9 8
Z ~ x x x x Z T x
d. ~ c, c. ca c~., c.
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
46
z
..
m x x x x x x x
a
oc o 0 0 0 0 0 0 0
~
s ~ _, x ,.
V ~, ~1
04 ,H a '~ V ~ U U
en
n
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z
._
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V H ~ V H ~ V H ~ V H ~ U N ~ V H ~ U H ~ U H ~
i,
,a,8 ~ >,8 ~ ?~~'°- a,8 °- ~~'~ i,8 ~ .~,5 ~ T3
U ca ;G U ld ;C7 U N ~7 v nl ;C V (d :O V N Z7 U N 'n
U ~
~r
L~ M a CZi M ~ M v ~ M ...n ~ M ~ CG M yr G~ M a.i ~" M 'r
i ~ n v i n n
.a .g, .9 .e '~ .e
U U U U ~ U a U
b. ~, ~, a ~ ~ d.
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
47
~~.
..
x x x x x ~ x
II
a
s x x x
oC o 0
~
, 0 0 0 0
U
N U N
C C '
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c o U ~
~' Z
. . x
~ _ ~
V E ~ ~ E
_ U .
e
z
z
x
U H U H U H 1i! ' C
.-~. ~ ~"' ~
i, ' ~
.
VNC ~'sC VcSV~ Verlb >,s C
U N N v ~
'b ' '
~ _ U G
~ C ~ U U
~
04 11 II II II o vi a,
o ~ o o o G p U , ,
o II II
~
OG ~0~, C~ GY GY, CG 4 ~' V
i ~ 0. ~ p
0. G
OC 04 04 CG 04 p4 _
eh r, en eh en O
Ga C~, ~ v V ~ V E
e G
r
v
.6 -
cc ~ z x x
r:. ~ ~. ~ 4 d,
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
48
x
z
..
x x x x x x x x
a
a
ac o 0 0 0 0 0 0 0
N H
U U ~. :: '. '.
~ -~ .a
a, a, U U 2 T ~ Z
~9' . ~ .. ~. U U U U
i
z ~Z z z
E E ~ ~ ~ O O O
U U U U
a'
a. a. ~ ~ i, i,
o ~ ~ ~ v
a a ~ ~ a, c.
~4
N N ~ N ~ H
E~. ''~'Ea. ''~'Ea, '"Ei, '=Ea, '-'E>, '-'ET ''-'E>,
,a,9 ° a,3 E a,8 = a,s ~ ~3 E .>,°~~ ,i,8 E r8 E
V cd b V e0 ;b V ed ;fl V of b V etf 'C V c0 b V ~C 'G
V ~ ,~
0. x ~ O. OC ~ 0. ~ ~ 0. ~ d OC O~'. OG ~ G~'. 04 ~'~' 0.
04~~ ~r,~ ~~~,d acMd ocr,~ acr,~ xr,~ ocr,d
-e 'e -a ~ L -a o ~ -~ o a
~~ ~~ ~ ~ a. ~~ ~ a ~ a.
x U U U ~~ "Jr"~' U ~C ~ U
C ~,~, ~ a~ ~S. Y? e~ C ~ a
ri E c~~ E r~ E
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
49
x x x x x x x
a
a
oc o 0 0 0 0 0 0
z , _~.
U >, ~, ~'
-a ~
N
x x
U U T vi .~ ~ U
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...
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.a, 8 .E >, 9 E .~ 8 .E '~ 8 .E .a, 9 " .~ g ,c
v ~:v v cd;v v ~a;o v ~;v v coy v ~o:v
p4 II ° ~ II ° ~ x il ~ ~ 11 ° ~ II ~ ~ II
04 ~ a oG ~ 0. ~ ~ a x ~ ~ oG ~ a oG ~ 10''..
LY, M ~, ~ M V CX M ~ i~ M "'v~ L1C tit '~ C>r M ~
$ ~ $ '$ $ $ '-0
M n
h eu
d. 5 ~ b. ~ c, d, d.
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CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
,
04
Z
..
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a
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U N ..~., U N ~ U '~ ~ U of ~ ~ ,n ~ ,n ~ U N ~ U N ~~~~
E a, . E a, . E- i, - E T . E >, . E a, . E i, . E T
.a,8 ~ a,8 °- >,8 c ,>,g.G ~,B.C >,8 ° i,s ~c ~3
U td 'D v cb 'D V e~ :C
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OG
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Q ~ ~ ~. ~ ~ ~,
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
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CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
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CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
53
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CA 02290748 1999-11-19
WO 99/06437 PCT/US98116070
54
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a
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CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
x x x x x x x x
a
a
Q ~ q o o ~ o
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a
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CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
DETAILED DESCRIPTION OF THE INVENTION
As above, this invention relates to compounds which inhibit
leukocyte adhesion and, in particular, leukocyte adhesion mediated by VLA-
4. However, prior to describing this invention in further detail, the
following
terms will first be defined.
Definitions
As used herein, "alkyl" refers to alkyl groups preferably having from
1 to 10 carbon atoms and more preferably 1 to 6 carbon atoms. This term is
10 exemplified by groups such as methyl, t-butyl, n-heptyl, octyl and the
like.
"Substituted alkyl" refers to an alkyl group, preferably of from 1 to
10 carbon atoms, having from 1 to S substituents selected from the group
consisting of alkoxy, substituted alkoxy, acyl, acylamino,
15 thiocarbonylamino, acyloxy, amino, amidino, alkyl amidino,thioamidino,
aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,
aryloxylaryl, substituted aryloxyaryl, cyano, halogen, hydroxyl, nitro,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
20 carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,
guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,
thioaryl,
substituted thioaryl, thiocycloalkyl, substituted thiocycloalkyl,
25 thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted
thioheterocyclic, heteroaryl, substituted aryl, substituted heteroaryl,
heterocyclic, substituted heterocyclic, cycloalkoxy, substituted cycloalkoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino,
CA 02290748 1999-11-19
WO 99/06437 PCT/US98/16070
96
-OS(O)2-alkyl, -OS(O)2-substituted alkyl, -OS(O)2-aryl, -OS(O)z-substituted
aryl, -OS(O)2-heteroaryl, -OS(O)z-substituted heteroaryl, -OS(O)z-
heterocyclic, -OS(O)2-substituted heterocyclic, -OSOZ-NRR,
-NRS(O)2-alkyl, -NRS(O)Z-substituted alkyl, -NRS(O)2-aryl, -NRS(O)2-
substituted aryl, -NRS(O)2-heteroaryl, -NRS(O)2-substituted heteroaryl,
-NRS(O)~-heterocyclic, -NRS(O)z-substituted heterocyclic, -NRS(O)2-NR-
alkyl, -NRS(O)2-NR-substituted alkyl, -NRS(O)2-NR-aryl, -NRS(O)2-NR-
substituted aryl, -NRS(O)z-NR-heteroaryl, -NRS(O)z-NR-substituted
heteroaryl, -NRS(O)2-NR-heterocyclic, -NRS(O)2-NR-substituted
heterocyclic, mono- and di-alkylamino, mono- and di-(substituted
alkyl)amino, mono- and di-arylamino, mono- and di-(substituted aryi)amino,
mono- and di-heteroarylamino, mono- and di-(substituted heteroaryl)amino,
mono- and di-heterocyclic amino, mono- and di-(substituted heterocyclic)
amino, unsymmetric di-substituted amines having different substituents
1 S selected from the group consisting of alkyl, substituted alkyl, aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic; substituted alkyl groups having amino groups
blocked by conventional blocking groups (such as Boc, Cbz, formyl, and the
like) and alkyl/substituted alkyl groups substituted with -SO~-alkyl, -SOZ-
substituted alkyl, -SO,-alkenyl, -SO~-substituted alkenyl, -SO~-cycloalkyl,
-SOz-substituted cycloalkyl, -SOZ-aryl, -SO,-substituted aryl, -SOz-
heteroaryl, -SOZ-substituted heteroaryl, -SO,-heterocyclic, -SO~-substituted
heterocyclic or -SOZNRR, where R is hydrogen or alkyl.
"Alkoxy" refers to the group "alkyl-O-" which includes, by way of
example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, tert-butoxy,
sec-butoxy, n-pentoxy, n-hexoxy, 1,2-dimethylbutoxy, and the like.
"Substituted alkoxy" refers to the group "substituted alkyl-O-".
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"Acyl" refers to the groups H-C(O)-, alkyl-C(O)-, substituted alkyl-
C(O)-, alkenyl-C(O)-, substituted alkenyl-C(O)-, alkynyl-C(O)-, substituted
alkynyl-C(O)- cycloalkyl-C(O)-, substituted cycloalkyl-C(O)-, aryl-C(O)-,
substituted aryl-C(O)-, heteroaryl-C(O)-, substituted heteroaryl-C(O),
heterocyclic-C(O)-, and substituted heterocyclic-C(O)- wherein alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl,
substituted heteroaryl, heterocyclic and substituted heterocyclic are as
defined herein.
"Acylamino" refers to the group -C(O)NRR where each R is
independently selected from the group consisting of hydrogen, alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl,
substituted heteroaryl, heterocyclic, and substituted heterocyclic; and where
each R can be joined to form, together with the nitrogen atom, a heterocyclic
or substituted heterocyclic ring wherein alkyl, substituted alkyl, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic are as defined herein.
"Thiocarbonylamino" refers to the group -C(S)NRR where each R is
independently selected from the group consisting of hydrogen, alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl,
substituted heteroaryl, heterocyclic, and substituted heterocyclic, and where
each R can be joined to form, together with the nitrogen atom, a heterocyclic
or substituted heterocyclic ring wherein alkyl, substituted alkyl, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted
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cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic are as defined herein.
"Acyloxy" refers to the groups alkyl-C(O)O-, substituted alkyl-
s C(O)O-, alkenyl-C(O)O-, substituted alkenyl-C(O)O-, alkynyl-C(O)O-,
substituted alkynyl-C(O)O-, aryl-C(O)O-, substituted aryl-C(O)O-,
cycloalkyl-C(O)O-, substituted cycloalkyl-C(O)O-, heteroaryl-C(O)O-,
substituted heteroaryl-C(O)O-, heterocyclic-C(O)O-, and substituted
heterocyclic-C(O)O- wherein alkyl, substituted alkyl, alkenyl, substituted
alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,
aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic are as defined herein.
"Alkenyl" refers to alkenyl group preferably having from 2 to 10
carbon atoms and more preferably 2 to 6 carbon atoms and having at least 1
and preferably from 1-2 sites of alkenyl unsaturation.
"Substituted alkenyl" refers to alkenyl groups having from 1 to 5
substituents selected from the group consisting of alkoxy, substituted alkoxy,
acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino,
thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,
aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,
guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,
thioaryl,
substituted thioaryl, thiocycloalkyl, substituted thiocycloalkyl,
thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted
*rB
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thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyclic,
substituted
heterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino; -OS(O),-alkyl, -OS(O)2-
substituted alkyl, -OS(O)z-aryl, -OS(O)Z-substituted aryl, -OS(O)z-heteroaryl,
-OS(O)z-substituted heteroaryl, -OS(O)z-heterocyclic, -OS(O)2-substituted
heterocyclic, -OSO,-NRR, -NRS(O)2-alkyl, -NRS(O)2-substituted alkyl,
-NRS(O)z-aryl, -NRS(O)2-substituted aryl, -NRS(O),-heteroaryl, -NRS(O),-
substituted heteroaryl, -NRS(O)2-heterocyclic, -NRS(O),-substituted
heterocyclic, -NRS(O)z-NR-alkyl, -NRS(O)Z-NR-substituted alkyl,
-NRS(O),-NR-aryl, -NRS(O)Z-NR-substituted aryl, -NRS(O)2-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O)~-NR-heterocyclic,
-NRS(O),-NR-substituted heterocyclic; mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryI)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic and substituted alkenyl groups
having amino groups blocked by conventional blocking groups (such as Boc,
Cbz, formyl, and the like) and alkenyl/substituted alkenyl groups substituted
with -SO~-alkyl, -SOZ-substituted alkyl, -SO~-alkenyl, -SO,-substituted
alkenyl, -SO,-cycloalkyl, -SOz-substituted cycloalkyl, -SO~-aryl, -SOZ-
substituted aryl, -SOZ-heteroaryl, -SOZ-substituted heteroaryl, -SOz-
heterocyclic, -SO,-substituted heterocyclic or -SO,NRR, where R is
hydrogen or alkyl.
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"Alkynyl" refers to alkynyl group preferably having from 2 to 10
carbon atoms and more preferably 3 to 6 carbon atoms and having at least 1
and preferably from 1-2 sites of alkynyl unsaturation.
"Substituted alkynyl" refers to alkynyl groups having from 1 to 5
substituents selected from the group consisting of alkoxy, substituted alkoxy,
acyl, acylamino, thiocarbonylamino, acyloxy, amino, amidino, alkylamidino,
thioamidino, aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,
aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxyiheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,
1 S guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,
thioaryl,
substituted thioaryl, thiocycloalkyl, substituted thiocycloalkyl,
thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted
thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyclic,
substituted
heterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylarnino, oxythiocarbonylamino, -OS(O),-alkyl, -OS(O)2-
substituted alkyl, -OS(O)2-aryl, -OS(O)2-substituted aryl, -OS(O)2-heteroaryl,
-OS(O),-substituted heteroaryl, -OS(O)~-heterocyclic, -OS(O)~-substituted
heterocyclic, -OSO,-NRR, -NRS(O)2-alkyl, -NRS(O)~-substituted alkyl,
-NRS(O)~-aryl, -NRS(O)2-substituted aryl, -NRS(O)2-heteroaryl, -NRS(O),-
substituted heteroaryl, -NRS(O)2-heterocyclic, -NRS(O),-substituted
heterocyclic, -NRS(O)2-NR-alkyl, -NRS{O)z-NR-substituted alkyl,
-NRS(O),-NR-aryl, -NRS(O),-NR-substituted aryl, -NRS(O)z-NR-
heteroaryl, -NRS(O),-NR-substituted heteroaryl, -NRS(O),-NR-heterocyclic,
-NRS(O)z-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
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di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic; substituted alkynyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like), and alkynyl/substituted alkynyl groups substituted with
-SOZ-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SO,-substituted alkenyl,
-SOZ-cycloalkyl, -SO~-substituted cycloalkyl, -SOZ-aryl, -SOZ-substituted
aryl, -SOZ-heteroaryl, -S02-substituted heteroaryl, -SO~-heterocyclic, -SO,-
substituted heterocyclic or -SOZNRR, where R is hydrogen or alkyl.
"Amidino" refers to the group HZNC- and the term "alkylamidino"
NH
refers to compounds having I to 3 alkyl groups (e.g., alkylHNC-),
NH
"Thioamidino" refers to the group RSC- where R is hydrogen or
NH
alkyl.
"Aminoacyl" refers to the groups -NRC(O)alkyl,
-NRC(O)substituted alkyl, -NRC(O)cycloalkyl, -NRC(O)substituted
cycloalkyl, -NRC(O)alkenyl, -NRC{O)substituted alkenyl,
-NRC(O)alkynyl, -NRC(O)substituted alkynyl, -NRC(O)aryl,
-NRC(O)substituted aryl, -NRC(O)heteroaryl, -NRC(O)substituted
heteroaryl, -NRC(O)heterocyclic, and -NRC(O)substituted heterocyclic,
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where R is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic are as defined herein.
"Aminocarbonyloxy" refers to the groups -NRC(O)O-alkyl,
-NRC{O)O-substituted alkyl, -NRC(O)O-alkenyl, -NRC(O)O-substituted
alkenyl, -NRC(O)O-alkynyl, -NRC(O)O-substituted alkynyl, -NRC(O)O-
cycloalkyl, -NRC(O)O-substituted cycloalkyl, -NRC(O)O-aryl, -NRC(O)O-
substituted aryl, -NRC(O)O-heteroaryl, -NRC(O)O-substituted heteroaryl,
-NRC(O)O-heterocyclic, and -NRC(O)O-substituted heterocyclic where R is
hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl, substituted
alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl,
aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
I S substituted heterocyclic are as defined herein.
"Oxycarbonylamino" refers to the groups -OC(O)NRR,
-OC(O)NR-alkyl, -OC(O)NR-substituted alkyl, -OC(O)NR-alkenyl,
-OC(O)NR-substituted alkenyl, -OC(O)NR-alkynyl, -OC(O)NR-substituted
alkynyl, -OC(O)NR-cycloalkyl, -OC(O)NR-substituted cycloalkyl,
-OC(O)NR-aryl, -OC(O)NR-substituted aryl, -OC(O)NR-heteroaryl,
-OC(O)NR-substituted heteroaryl,- OC(O)NR-heterocyclic, and
-OC(O)NR-substituted heterocyclic where R is hydrogen or alkyl, and where
each R can be joined to form, together with the nitrogen atom, a heterocyclic
or substituted heterocyclic ring and wherein alkyl, substituted alkyl,
alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic are as defined herein.
"Oxythiocarbonylamino" refers to the groups -OC(S)NRR,
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-OC(S)NR-alkyl, -OC(S)NR-substituted alkyl, -OC(S)NR-alkenyl,
-OC(S)NR-substituted alkenyl, -OC(S)NR-alkynyl, -OC(S)NR-substituted
alkynyl, -OC(S)NR-cycloalkyl, -OC(S)NR-substituted cycloalkyl,
-OC(S)NR-aryl, -OC(S)NR-substituted aryl, -OC(S)NR-heteroaryl,
-OC(S)NR-substituted heteroaryl, -OC(S)NR-heterocyciic, and -OC(S)NR-
substituted heterocyclic where R is hydrogen or alkyl, or where each R can
be joined to form, together with the nitrogen atom, a heterocyclic or
substituted heterocyclic ring and wherein alkyl, substituted alkyl, alkenyl,
substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted
cycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic are as defined herein.
"Aminocarbonylamino" refers to the groups -NRC(O)NRR,
-NRC(O)NR-alkyl, -NRC(O)NR-substituted alkyl, -NRC(O)NR-alkenyl,
-NRC(O)NR-substituted alkenyl, -NRC(O)NR-alkynyl, -NRC(O)NR-
substituted alkynyl, -NRC(O)NR-aryl, -NRC(O)NR-substituted aryl,
-NRC(O)NR-cycloalkyl, -NRC(O)NR-substituted cycloalkyl, -NRC(O)NR-
heteroaryl, -NRC(O)NR-substituted heteroaryl, -NRC(O)NR-heterocyclic,
and -NRC(O)NR-substituted heterocyclic where each R is independently
hydrogen, alkyl, and where each R can be joined to form, together with the
nitrogen atom, a heterocyclic or substituted heterocyclic ring as well as
where one of the amino groups is blocked by conventional blocking groups
(such as Boc, Cbz, formyl, and the like) and wherein alkyl, substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, substituted aryl, heteroaryl, substituted
heteroaryl, heterocyclic and substituted heterocyclic are as defined herein.
"Aminothiocarbonylamino" refers to the groups -NRC(S)NRR,
-NRC(S)NR-alkyl, -NRC(S)NR-substituted alkyl, -NRC(S)NR-alkenyl,
*rB
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-NRC(S)NR-substituted alkenyl, -NRC(S)NR-alkynyl, -NRC(S)NR-
substituted alkynyl, -NRC(S)NR-aryl, -NRC(S)NR-substituted aryl,
-NRC(S)NR-cycloalkyl, -NRC(S)NR-substituted cycloalkyl, -NRC(S)NR-
heteroaryl, and -NRC(S)NR-substituted heteroaryl, -NRC(S)NR-
heterocyclic, and -NRC(S)NR-substituted heterocyclic where each R is
independently hydrogen, alkyl, and where each R can be joined to form,
together with the nitrogen atom, a heterocyclic or substituted heterocyclic
ring as well as where one of the amino groups is blocked by conventional
blocking groups (such as Boc, Cbz, formyl, and the like) and wherein alkyl,
substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,
cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl,
substituted heteroaryl, heterocyclic and substituted heterocyclic are as
defined herein.
"Aryl" or "Ar" refers to an unsaturated aromatic carbocyclic group of
from 6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiple
condensed rings (e.g., naphthyl or anthryl) which condensed rings may or
may not be aromatic (e.g., 2-benzoxazolinone, 2H-1,4-benzoxazin-3(4H)-
one-7-yl, and the like). Preferred aryls include phenyl and naphthyl.
Substituted aryl refers to aryl groups which are substituted with from
1 to 3 substituents selected from the group consisting of hydroxy, acyl,
acylamino, thiocarbonylamino, acyloxy, alkyl, substituted alkyl, alkoxy,
substituted alkoxy, alkenyl, substituted alkenyl, alkynyl, substituted
alkynyl,
amidino, alkylamidino, thioamidino, amino, aminoacyl, aminocarbonyloxy,
aminocarbonylamino, aminothiocarbonylamino, aryl, substituted aryl,
aryloxy, substituted aryloxy, cycloalkoxy, substituted cycloalkoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted alkyl,
carboxyl-cycloalkyl, carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-
*rB
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substituted aryl, carboxylheteroaryl, carboxyl-substituted heteroaryl,
carboxylheterocyclic, carboxyl-substituted heterocyclic, carboxylamido,
cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl, substituted
thioaryl,
thioheteroaryl, substituted thioheteroaryl, thiocycloalkyl, substituted
thiocycloalkyl, thioheterocyclic, substituted thioheterocyclic, cycloalkyl,
substituted cycloalkyl, guanidino, guanidinosulfone, halo, nitro, heteroaryl,
substituted heteroaryl, heterocyclic, substituted heterocyclic, cycloalkoxy,
substituted cycloalkoxy, heteroaryloxy, substituted heteroaryloxy,
heterocyclyloxy, substituted heterocyclyloxy, oxycarbonylamino,
oxythiocarbonylamino, -S(O),-alkyl, -S(O)S-substituted alkyl, -S(O)2-
cycloalkyl, -S(O)S-substituted cycloalkyl, -S(O),-alkenyl, -S(O)2-substituted
alkenyl, -S(O)2-aryl, -S(O)2-substituted aryl, -S(O),-heteroaryl, -S(O)2-
substituted heteroaryl, -S{O),-heterocyclic, -S(O),-substituted heterocyclic,
-OS(O)z-alkyl, -OS(O),-substituted alkyl, -OS(O),-aryl, -OS(O),-substituted
aryl, -OS(O)2-heteroaryl, -OS(O),-substituted heteroaryl, -OS(O)z-
heterocyclic, -OS(O)~-substituted heterocyclic, -OSO.,-NRR, -NRS{O)Z-
alkyl, -NRS(O),-substituted alkyl, -NRS(O)2-aryl, -NRS(O),-substituted aryl,
-NRS(O),-heteroaryl, -NRS(O)Z-substituted heteroaryl, -NRS(O)Z-
heterocyclic, -NRS(O)~-substituted heterocyclic, -NRS(O),-NR-alkyl,
-NRS(O),-NR-substituted alkyl, -NRS(O),-NR-aryl, -NRS(O),-NR-
substituted aryl, -NRS(O)~-NR-heteroaryl, -NRS(O),-NR-substituted
heteroaryl, -NRS(O),-NR-heterocyclic, -NRS(O),-NR-substituted
heterocyclic, mono- and di-alkylamino, mono- and di-(substituted
alkyl)amino, mono- and di-arylamino, mono- and di-(substituted aryl)amino,
mono- and di-heteroarylamino, mono- and di-(substituted heteroaryl)amino,
mono- and di-heterocyclic amino, mono- and di-(substituted heterocyclic)
amino, unsymmetric di-substituted amines having different substituents
selected from the group consisting of alkyl, substituted alkyl, aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic,
substituted
heterocyclic, amino groups on the substituted aryl blocked by conventional
*rB
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blocking groups (such as Boc, Cbz, formyl, and the like), and -SOZNRR,
where R is hydrogen or alkyl.
"Aryloxy" refers to the group aryl-O- which includes, by way of
example, phenoxy, naphthoxy, and the like.
"Substituted aryloxy" refers to substituted aryl-O- groups.
"Aryloxyaryl" refers to the group -aryl-O-aryl.
"Substituted aryloxyaryl" refers to aryloxyaryl groups substituted
with from 1 to 3 substituents on either or both aryl rings selected from the
group consisting of hydroxy, acyl, acylamino, thiocarbonylamino, acyloxy,
alkyl, substituted alkyl, aikoxy, substituted alkoxy, alkenyl, substituted
alkenyl, alkynyl, substituted alkynyl, amidino, alkylamidino, thioamidino,
amino, aminoacyl, aminocarbonyloxy, aminocarbonylamino,
aminothiocarbonylamino, aryl, substituted aryl, aryloxy, substituted aryloxy,
cycloalkoxy, substituted cycloalkoxy, heteroaryloxy, substituted
heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy, carboxyl,
carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl, carboxyl-
substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, carboxylamido, cyano, thiol, thioalkyl,
substituted thioalkyl, thioaryl, substituted thioaryl, thioheteroaryl,
substituted
thioheteroaryl, thiocycloalkyl, substituted thiocycloalkyl, thioheterocyclic,
substituted thioheterocyclic, cycloalkyl, substituted cycloalkyl, guanidino,
guanidinosulfone, halo, nitro, heteroaryl, substituted heteroaryl,
heterocyclic,
substituted heterocyclic, cycloalkoxy, substituted cycloalkoxy,
heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy, substituted
heterocyclyloxy, oxycarbonylamino, oxythiocarbonylamino,
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-S(O)z-alkyl, -S(O)2-substituted alkyl, -S(O),-cycloalkyl, -S(O)2-substituted
cycloalkyl, -S(O)2-alkenyl, -S(O)2-substituted alkenyl, -S(O)z-aryl, -S(O)Z-
substituted aryl, -S(O)2-heteroaryl, -S(O)z-substituted heteroaryl, -S(O)Z-
heterocyclic, -S(O)z-substituted heterocyclic, -OS(O)z-alkyl, -OS(O)2-
substituted alkyl, -OS(O)2-aryl, -OS(O)Z-substituted aryl, -OS(O)2-heteroaryl,
-OS(O)Z-substituted heteroaryl, -OS(O)z-heterocyclic, -OS(O)Z-substituted
heterocyclic, -OSOz-NRR, -NRS(O)2-alkyl, -NRS(O)z-substituted alkyl,
-NRS(O),-aryl, -NRS(O)z-substituted aryl, -NRS(O),-heteroaryl, -NRS(O)2-
substituted heteroaryl, -NRS(O)2-heterocyclic, -NRS(O)2-substituted
heterocyclic, -NRS(O)2-NR-alkyl, -NRS(O)2-NR-substituted alkyl,
-NRS(O)Z-NR-aryl, -NRS(O)~-NR-substituted aryl, -NRS(O)2-NR-
heteroaryi, -NRS(O),-NR-substituted heteroaryl, -NRS(O)2-NR-heterocyclic,
-NRS(O)~-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic, amino groups on the substituted
aryl blocked by conventional blocking groups (such as Boc, Cbz, formyl, and
the like) and substituted with -SO,NRR, where R is hydrogen or alkyl.
"Cycloalkyl" refers to cyclic alkyl groups of from 3 to 8 carbon atoms
having a single cyclic ring including, by way of example, cyclopropyl,
cyclobutyl, cyclopentyl, cyclooctyl and the like. Excluded from this
definition are mufti-ring alkyl groups such as adamantanyl, etc.
"Cycloalkenyl" refers to cyclic alkenyl groups of from 3 to 8 carbon
atoms having single or multiple unsaturation but which are not aromatic.
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"Substituted cycloalkyl" and "substituted cycloalkenyl" refer to a
cycloalkyl and cycloalkenyl groups, preferably of from 3 to 8 carbon atoms,
having from 1 to 5 substituents selected from the group consisting of oxo
(=O), thioxo (=S), alkoxy, substituted alkoxy, acyl, acylamino,
thiocarbonylamino, acyloxy, amino, amidino, alkylamidino, thioamidino,
aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,
aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, vitro,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,
guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,
thioaryl,
substituted thioaryl, thiocycloalkyl, substituted thiocycloalkyl,
thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted
thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyclic,
substituted
heterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -OS(O),-alkyl, -OS(O),-
substituted alkyl, -OS(O)2-aryl, -OS(O)~-substituted aryl, -OS(O)Z-heteroaryl,
-OS(O),-substituted heteroaryl, -OS(O)Z-heterocyclic, -OS(O),-substituted
heterocyclic, -OSOZ-NRR, -NRS(O)2-alkyl, -NRS(O),-substituted alkyl,
-NRS(O)~-aryl, -NRS(O)Z-substituted aryl, -NRS(O),-heteroaryl, -NRS(O),-
substituted heteroaryl, -NRS(O),-heterocyclic, -NRS(O)~-substituted
heterocyclic, -NRS(O),-NR-alkyl, -NRS(O)~-NR-substituted alkyl,
-NRS{O),-NR-aryl, -NRS(O)Z-NR-substituted aryl, -NRS(O)~-NR-
heteroaryl, -NRS(O),-NR-substituted heteroaryl, -NRS(O),-NR-heterocyclic,
-NRS(O),-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
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(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryi,
S heterocyclic and substituted heterocyclic, substituted alkynyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
formyl, and the like) and alkynyl/substituted alkynyl groups substituted with
-SOZ-alkyl, -SOZ-substituted alkyl, -SOZ-alkenyl, -SO~-substituted alkenyl, -
SOz-cycloalkyl, -SOZ-substituted cycloalkyl, -SOz-aryl, -SOZ-substituted aryl,
IO -SO,-heteroaryl, -SOz-substituted heteroaryl, -SOZ-heterocyclic, -SOZ-
substituted heterocyclic or -SOZNRR, where R is hydrogen or alkyl.
"Cycloalkoxy" refers to -O-cycloalkyl groups.
1 S "Substituted cycloalkoxy" refers to -O-substituted cycloalkyl groups.
"Guanidino" refers to the groups -NRC(=NR)NRR,
-NRC(=NR)NR-alkyl, -NRC(=NR)NR-substituted alkyl,
-NRC(=NR)NR-alkenyl, -NRC(=NR)NR-substituted alkenyl,
20 -NRC(=NR)NR-alkynyl, -NRC(=NR)NR-substituted alkynyl,
-NRC(=NR)NR-aryl, -NRC(=NR)NR-substituted aryl, -NRC(=NR)NR-
cycloalkyl, -NRC(=NR)NR-substituted cycloalkyl,-NRC(=NR)NR-
heteroaryl, -NRC(=NR)NR-substituted heteroaryl, -NRC(=NR)NR-
heterocyclic, and -NRC(=NR)NR-substituted heterocyclic where each R is
2S independently hydrogen and alkyl as well as where one of the amino groups
is blocked by conventional blocking groups (such as Boc, Cbz, formyl, and
the like) and wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl,
alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
30 substituted heterocyclic are as defined herein.
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"Guanidinosulfone" refers to the groups -NRC(=NR)NRSOz-alkyl,
-NRC(=NR)NRSOZ-substituted alkyl, -NRC(=NR)NRSOZ-alkenyl,
-NRC(=NR)NRSOZ-substituted alkenyl, -NRC(=NR)NRSO~-alkynyl,
-NRC(=NR)NRSOZ-substituted alkynyl, -NRC(=NR)NRSOz-aryl,
-NRC(=NR)NRSOZ-substituted aryl, -NRC(=NR)NRSO~-cycloalkyl,
-NRC(=NR)NRSOz-substituted cycloalkyl, -NRC(=NR)NRSOZ-heteroaryl,
and -NRC(=NR)NRSOZ-substituted heteroaryl, -NRC(=NR)NRSOZ-
heterocyclic, and -NRC(=NR)NRSOZ-substituted heterocyclic where each R
is independently hydrogen or alkyl and wherein alkyl, substituted alkyl,
alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,
substituted cycloalkyl, aryl, substituted aryl, heteroaryi, substituted
heteroaryl, heterocyclic and substituted heterocyclic are as defined herein.
"Halo" or "halogen" refers to fluoro, chloro, bromo and iodo and
preferably is either chloro or bromo.
"Heteroaryl" refers to an aromatic carbocyclic group of from 2 to 10
carbon atoms and 1 to 4 heteroatoms selected from the group consisting of
oxygen, nitrogen and sulfur within the ring. Such heteroaryl groups can have
a single ring (e.g., pyridyl or furyl) or multiple condensed rings (e.g.,
indolizinyl or benzothienyl). Preferred heteroaryls include pyridyl, pyrrolyl,
indolyl and filryl.
"Substituted heteroaryl" refers to heteroaryl groups which are
substituted with from 1 to 3 substituents selected from the group consisting
of hydroxy, acyl, acylamino, thiocarbonylamino, acyloxy, alkyl, substituted
alkyl, alkoxy, substituted alkoxy, alkenyl, substituted alkenyl, alkynyl,
substituted alkynyl, amidino, alkylamidino, thioamidino, amino, aminoacyl,
aminocarbonyloxy, aminocarbonylamino, aminothiocarbonylamino, aryl,
substituted aryl, aryloxy, substituted aryloxy, cycloalkoxy, substituted
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cycloalkoxy, heteroaryloxy, substituted heteroaryloxy, heterocyclyloxy,
substituted heterocyclyloxy, carboxyl, carboxylalkyl, carboxyl-substituted
alkyl, carboxyl-cycloaikyl, carboxyl-substituted cycloalkyl, carboxylaryl,
carboxyl-substituted aryl, carboxylheteroaryl, carboxyl-substituted
heteroaryl, carboxylheterocyclic, carboxyl-substituted heterocyclic,
carboxylamido, cyano, thiol, thioalkyl, substituted thioalkyl, thioaryl,
substituted thioaryl, thioheteroaryl, substituted thioheteroaryl,
thiocycloalkyl,
substituted thiocycloalkyl, thioheterocyclic, substituted thioheterocyclic,
cycloalkyl, substituted cycloalkyl, guanidine, guanidinosulfone, halo, nitre,
heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic,
cycloalkoxy, substituted cycloalkoxy, heteroaryloxy, substituted
heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -S(O)S-alkyl, -S(O)2-substituted
alkyl, -S(O)2-cycloalkyl, -S(O)2-substituted cycloalkyl, -S(O)z-alkenyl,
-S(O)2-substituted alkenyl, -S(O)2-aryl, -S(O)S-substituted aryl, -S(O)z-
heteroaryl, -S(O)2-substituted heteroaryl, -S{O)Z-heterocyclic, -S(O)2-
substituted heterocyclic, -OS(O)2-alkyl, -OS(O)z-substituted alkyl, -OS(O),-
aryl, -OS(O)2-substituted aryl, -OS(O)2-heteroaryl, -OS{O)2-substituted
heteroaryl, -OS(O)z-heterocyclic, -OS(O)2-substituted heterocyclic, -OSOZ-
NRR, -NRS(O),-alkyl, -NRS(O)2-substituted alkyl, -NRS(O),-aryl,
-NRS(O),-substituted aryl, -NRS(O),-heteroaryl, -NRS(O),-substituted
heteroaryl, -NRS(O),-heterocyclic, -NRS(O)z-substituted heterocyclic,
-NRS(O),-NR-alkyl, -NRS(O)Z-NR-substituted alkyl, -NRS(O)2-NR-aryl,
-NRS(O),-NR-substituted aryl, -NRS(O)Z-NR-heteroaryl, -NRS(O)~-NR-
substituted heteroaryl, -NRS(O)Z-NR-heterocyclic, -NRS(O)z-NR-substituted
heterocyclic, mono- and di-alkylamino, mono- and di-(substituted
alkyl)amino, mono- and di-arylamino, mono- and di-(substituted aryl)amino,
mono- and di-heteroarylamino, mono- and di-(substituted heteroaryl)amino,
mono- and di-heterocyclic amino, mono- and di-(substituted heterocyclic)
amino, unsymmetric di-substituted amines having different substituents
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selected from the group consisting of alkyl, substituted alkyl, aryl,
substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and
substituted heterocyclic, amino groups on the substituted aryl blocked by
conventional blocking groups (such as Boc, Cbz, formyl, and the like), and
-SOZNRR, where R is hydrogen or alkyl.
"Heteroaryloxy" refers to the group -O-heteroaryl and "substituted
heteroaryloxy" refers to the group -O-substituted heteroaryl.
"Heterocycle" or "heterocyclic" refers to a saturated or unsaturated
group having a single ring or multiple condensed rings, from 1 to 10 carbon
atoms and from 1 to 4 hetero atoms selected from the group consisting of
nitrogen, sulfur or oxygen within the ring wherein, in fused ring systems, one
or more the rings can be aryl or heteroaryl.
"Saturated heterocyclic" refers to heterocycles of single or multiple
condensed rings lacking unsaturation in any ring (e.g., carbon to carbon
unsaturation, carbon to nitrogen unsaturation, nitrogen to nitrogen
unsaturation, and the like).
"Unsaturated heterocyclic" refers to non-aromatic heterocycles of
single or multiple condensed rings having unsaturation in any ring (e.g.,
carbon to carbon unsaturation, carbon to nitrogen unsaturation, nitrogen to
nitrogen unsaturation, and the like).
"Substituted heterocyclic" refers to heterocycle groups which are
substituted with from 1 to 3 substituents selected from the group consisting
of oxo (=O), thioxo (=S), alkoxy, substituted alkoxy, acyl, acylamino,
thiocarbonylamino, acyloxy, amino, amidino, alkylamidino, thioamidino,
aminoacyl, aminocarbonylamino, aminothiocarbonylamino,
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aminocarbonyloxy, aryl, substituted aryl, aryloxy, substituted aryloxy,
aryloxyaryl, substituted aryloxyaryl, halogen, hydroxyl, cyano, nitro,
carboxyl, carboxylalkyl, carboxyl-substituted alkyl, carboxyl-cycloalkyl,
carboxyl-substituted cycloalkyl, carboxylaryl, carboxyl-substituted aryl,
carboxylheteroaryl, carboxyl-substituted heteroaryl, carboxylheterocyclic,
carboxyl-substituted heterocyclic, cycloalkyl, substituted cycloalkyl,
guanidino, guanidinosulfone, thiol, thioalkyl, substituted thioalkyl,
thioaryl,
substituted thioaryl, thiocycloalkyl, substituted thiocycloalkyl,
thioheteroaryl, substituted thioheteroaryl, thioheterocyclic, substituted
thioheterocyclic, heteroaryl, substituted heteroaryl, heterocyciic,
substituted
heterocyclic, cycloalkoxy, substituted cycloalkoxy, heteroaryloxy,
substituted heteroaryloxy, heterocyclyloxy, substituted heterocyclyloxy,
oxycarbonylamino, oxythiocarbonylamino, -OS(O)2-alkyl, -OS(O)2-
substituted alkyl, -OS(O)z-aryl, -OS(O)Z-substituted aryl, -OS(O)2-heteroaryl,
-OS(O)2-substituted heteroaryl, -OS(O)z-heterocyclic, -OS(O)z-substituted
heterocyclic, -OSOZ-NRR, -NRS(O),-alkyl, -NRS(O)~-substituted alkyl,
-NRS(O),-aryl, -NRS(O)z-substituted aryl, -NRS(O)Z-heteroaryl, -NRS(O)2-
substituted heteroaryl, -NRS(O)Z-heterocyclic, -NRS(O)2-substituted
heterocyclic, -NRS(O)2-NR-alkyl, -NRS(O)z-NR-substituted alkyl,
-NRS(O)2-NR-aryl, -NRS(O)Z-NR-substituted aryl, -NRS(O),-NR-
heteroaryl, -NRS(O)2-NR-substituted heteroaryl, -NRS(O),-NR-heterocyclic,
-NRS(O)Z-NR-substituted heterocyclic, mono- and di-alkylamino, mono- and
di-(substituted alkyl)amino, mono- and di-arylamino, mono- and di-
(substituted aryl)amino, mono- and di-heteroarylamino, mono- and di-
(substituted heteroaryl)amino, mono- and di-heterocyclic amino, mono- and
di-(substituted heterocyclic) amino, unsymmetric di-substituted amines
having different substituents selected from the group consisting of alkyl,
substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl,
heterocyclic and substituted heterocyclic, substituted alkynyl groups having
amino groups blocked by conventional blocking groups (such as Boc, Cbz,
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formyl, and the like) and alkynyl/substituted alkynyl groups substituted with
-SOZ-alkyl, -SOz-substituted alkyl, -SOz-alkenyl, -SOZ-substituted alkenyl,
-SOZ-cycloalkyl, -SOZ-substituted cycloalkyl, -SOZ-aryl, -SOZ-substituted
aryl, -SOZ-heteroaryl, -SOZ-substituted heteroaryl, -SOz-heterocyclic, -SOz-
substituted heterocyclic or -SOZNRR, where R is hydrogen or alkyl.
Examples of heterocycles and heteroaryls include, but are not limited
to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine,
pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine,
quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine,
quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline,
phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole,
phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine,
piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-
tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,
benzo[b]thiophene, morpholino, thiomorpholino, piperidinyl, pyrrolidine,
tetrahydrofuranyl, and the like.
"Saturated substituted heterocyclic" refers to substituted heterocycles
of single or multiple condensed rings lacking unsaturation in any ring (e.g.,
carbon to carbon unsaturation, carbon to nitrogen unsaturation, nitrogen to
nitrogen unsaturation, and the like).
"Unsaturated substituted heterocyclic" refers to non-aromatic
substituted heterocycles of single or multiple condensed rings having
unsaturation in any ring (e.g., carbon to carbon unsaturation, carbon to
nitrogen unsaturation, nitrogen to nitrogen unsaturation, and the like).
"Heterocyclyloxy" refers to the group -O-heterocyclic and
"substituted heterocyclyloxy" refers to the group -O-substituted heterocyclic.
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"Thiol" refers to the group -SH.
"Thioalkyl" refers to the groups -S-alkyl.
"Substituted thioalkyl" refers to the group -S-substituted alkyl.
"Thiocycloalkyl" refers to the groups -S-cycloalkyl.
"Substituted thiocycloalkyl" refers to the group -S-substituted
cycloalkyl.
"Thioaryl" refers to the group -S-aryl and "substituted thioaryl" refers
to the group -S-substituted aryl.
"Thioheteroaryl" refers to the group -S-heteroaryl and "substituted
thioheteroaryl" refers to the group -S-substituted heteroaryl.
"Thioheterocyclic" refers to the group -S-heterocyclic and
"substituted thioheterocyclic" refers to the group -S-substituted
heterocyclic.
"Pharmaceutically acceptable salt" refers to pharmaceutically
acceptable salts of a compound of Formula I or Formula IA, which salts are
derived from a variety of organic and inorganic counter ions well known in
the art and include, by way of example only, sodium, potassium, calcium,
magnesium, ammonium, tetraalkylammonium, and the like; and when the
compound (of Formula I or IA) contains a basic functionality, salts of
organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate,
mesylate, acetate, maleate, oxalate and the like.
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Comnoun_d Preparation
The compounds of this invention can be prepared from readily
available starting materials using the following general methods and
procedures. It will be appreciated that where typical or preferred process
conditions (i.e., reaction temperatures, times, mole ratios of reactants,
solvents, pressures, etc.) are given, other process conditions can also be
used
unless otherwise stated. Optimum reaction conditions may vary with the
particular reactants or solvent used, but such conditions can be determined by
one skilled in the art by routine optimization procedures.
Additionally, as will be apparent to those skilled in the art,
conventional protecting groups may be necessary to prevent certain
functional groups from undergoing undesired reactions. Suitable protecting
groups for various functional groups as well as suitable conditions for
protecting and deprotecting particular functional groups are well known in
the art. For example, numerous protecting groups are described in T. W.
Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, Second
Edition, Wiley, New York, 1991, and references cited therein.
Furthermore, the compounds of this invention will typically contain
one or more chiral centers. Accordingly, if desired, such compounds can be
prepared or isolated as pure stereoisomers, i.e., as individual enantiomers or
diastereomers, or as stereoisomer-enriched mixtures. All such stereoisomers
(and enriched mixtures) are included within the scope of this invention,
unless otherwise indicated. Pure stereoisomers (or enriched mixtures) may
be prepared using, for example, optically active starting materials or
stereoselective reagents well-known in the art. Alternatively, racemic
mixtures of such compounds can be separated using, for example, chiral
column chromatography, chiral resolving agents and the like.
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In a preferred method of synthesis, the compounds of formula I and
IA wherein Q is -C(O)NR'- are prepared by first coupling an amino acid of
formula II:
R3
RZ-NH-C-COOH II
1
H
wherein RZ and R3 are as defined above, with a sulfonyl chloride of formula
III:
R'-SOZ-Cl III
wherein R' is as defined above, to provide an N sulfonyl amino acid of
formula IV:
R3
R'-SOZ-N(RZ)-C-COOH IV
H
wherein R'-R3 are as defined above.
This reaction is typically conducted by contacting the amino acid of
formula II with at least one equivalent, preferably about 1.1 to about 2
equivalents, of sulfonyl chloride III in an inert diluent such as
dichloromethane and the like. Generally, the reaction is conducted at a
temperature ranging from about -70 ° C to about 40 ° C for about
1 to about 24
hours. Preferably, this reaction is conducted in the presence of a suitable
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base to scavenge the acid generated during the reaction. Suitable bases
include, by way of example, tertiary amines, such as triethylamine,
diisopropylethylamine, N methylmorpholine and the like. Alternatively, the
reaction can be conducted under Schotten-Baumann-type conditions using
aqueous alkali, such as sodium hydroxide and the like, as the base. Upon
completion of the reaction, the resulting N sulfonyl amino acid IV is
recovered by conventional methods including neutralization, extraction,
precipitation, chromatography, filtration, and the like.
The amino acids of formula II employed in the above reaction are
either known compounds or compounds that can be prepared from known
compounds by conventional synthetic procedures. Examples of suitable
amino acids for use in this reaction include, but are not limited to, L-
proline,
traps-4-hydroxyl-L-proline, cis-4-hydroxyl-L-proline, traps-3-phenyl-L-
proline, cis-3-phenyl-L-proline, L-(2-methyl)proline, L-pipecolinic acid, L-
azetidine-2-carboxylic acid,glycine, 2-tert-butylglycine, D,L-phenylglycine,
L-alanine, a-methylalanine, N methyl-L-phenylalanine, L-diphenylalanine,
sarcosine, D,L-phenylsarcosine, L-aspartic acid ~i-tert-butyl ester, L-
glutamic
acid 'y-tert-butyl ester, L-(O-benzyl)serine, 1-aminocyclopropanecarboxylic
acid, 1-aminocyclobutanecarboxylic acid, 1-aminocyclopentanecarboxylic
acid (cycloleucine) 1-aminocyclo-hexanecarboxylic acid, L-serine and the
like. If desired, the corresponding carboxylic acid esters of the amino acids
of formula II, such as the methyl esters, ethyl esters and the like, can be
employed in the above reaction with the sulfonyl chloride III. Subsequent
hydrolysis of the ester group to the carboxylic acid using conventional
reagents and conditions, i.e., treatment with an alkali metal hydroxide in an
inert diluent such as methanol/water, then provides the N sulfonyl amino
acid IV.
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Similarly, the sulfonyl chlorides of formula III employed in the above
reaction are either known compounds or compounds that can be prepared
from known compounds by conventional synthetic procedures. Such
compounds are typically prepared from the corresponding sulfonic acid, i.e.,
from compounds of the formula R'-S03H where R' is as defined above,
using phosphorous trichloride and phosphorous pentachloride. This reaction
is generally conducted by contacting the sulfonic acid with about 2 to 5
molar equivalents of phosphorous trichloride and phosphorous pentachloride,
either neat or in an inert solvent, such as dichloromethane, at temperature in
the range of about 0°C to about 80°C for about 1 to about 48
hours to afford
the sulfonyl chloride. Alternatively, the sulfonyl chlorides of formula III
can
be prepared from the corresponding thiol compound, i.e., from compounds of
the formula R'-SH where R' is as defined above, by treating the thiol with
chlorine (C12) and water under conventional reaction conditions.
Examples of sulfonyl chlorides suitable for use in this invention
include, but are not limited to, methanesulfonyl chloride, 2-propanesulfonyl
chloride, 1-butanesulfonyl chloride, benzenesulfonyl chloride,
1-naphthalenesulfonyl chloride, 2-naphthalenesulfonyl chloride,
p-toluenesulfonyl chloride, a-toluenesulfonyl chloride,
4-acetamidobenzenesulfonyl chloride, 4-amidinobenzenesulfonyl chloride,
4-tert-butylbenzenesulfonyl chloride, 4-bromobenzenesulfonyl chloride,
2-carboxybenzenesulfonyl chloride, 4-cyanobenzenesulfonyl chloride, 3,4-
dichlorobenzenesulfonyl chloride, 3,5-dichlorobenzenesulfonyl chloride, 3,4-
dimethoxybenzenesulfonyl chloride, 3,5-ditrifluoromethylbenzenesulfonyl
chloride, 4-fluorobenzenesulfonyl chloride, 4-methoxybenzenesulfonyl
chloride, 2-methoxycarbonylbenzenesulfonyl chloride,
4-methylamidobenzenesulfonyl chloride, 4-nitrobenzenesulfonyl chloride,
4-thioamidobenzenesulfonyl chloride, 4-trifluoromethylbenzenesulfonyl
chloride, 4-trifluoromethoxybenzenesulfonyl chloride, 2,4,6-
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trimethylbenzenesulfonyl chloride, 2-phenylethanesulfonyl chloride,
2-thiophenesulfonyl chloride, 5-chloro-2-thiophenesulfonyl chloride, 2,5-
dichloro-4-thiophenesulfonyl chloride, 2-thiazolesulfonyl chloride, 2-methyl-
4-thiazolesulfonyl chloride, 1-methyl-4-imidazolesulfonyl chloride,
1-methyl-4-pyrazolesulfony~ chloride, S-chloro-1,3-dimethyl-4-
pyrazolesulfonyl chloride, 3-pyridinesulfonyl chloride, 2-pyrimidinesulfonyl
chloride, and the like. If desired, a sulfonyl fluoride, sulfonyl bromide or
sulfonic acid anhydride may be used in place of the sulfonyl chloride in the
above reaction to form the N sulfonyl amino acids of formula IV.
The intermediate N sulfonyl amino acids of formula IV, wherein R3
is hydrogen, can also be prepared by reacting a sulfonamide of formula V:
R'-SOZ-NH-RZ V
wherein R' and RZ are as defined above, with a carboxylic acid derivative of
the formula L(R3)CHCOOR where L is a leaving group, such as chloro,
bromo, iodo, mesylate, tosylate and the like, R' is as defined above and R is
hydrogen or an alkyl group. This reaction is typically conducted by
contacting the sulfonamide V with at least one equivalent, preferably 1.1 to 2
equivalents, of the carboxylic acid derivative in the presence of a suitable
base, such as triethylamine, in an inert diluent, such as DMF, at a
temperature ranging from about 24 ° C to about 3 7 ° C for about
0.5 to about 4
hours. This reaction is further described in Zuckermann et al., J. Am. Chem.
Soc.,1992, 114, 10646-10647. Preferred carboxylic acid derivatives for use
in this reaction are a-chloro and a-bromocarboxylic acid esters such as tert-
butyl bromoacetate, and the like. When an carboxylic acid ester is employed
in this reaction, the ester group is subsequently hydrolyzed using
conventional procedures to afford an N sulfonyl amino acid of formula IV.
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The compounds of formula I and IA are then prepared by coupling
the intermediate N sulfonyl amino acid of formula IV with an amino acid
derivative of formula VI:
O
II
R'-NH-CH-C-R6 VI
Rs
wherein RS-R' are as defined above. This coupling reaction is typically
conducted using well-known coupling reagents such as carbodiimides, BOP
reagent (benzotriazol-1-yloxy-tris(dimethylamino)phosphonium
hexafluorophosphonate) and the like. Suitable carbodiimides include, by
way of example, dicyclohexylcarbodiimide (DCC), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and the like. If desired,
polymer supported forms of carbodiimide coupling reagents may also be
used including, for example, those described in Tetrahedron Letters, 34(48),
7685 (1993). Additionally, well-known coupling promoters, such as N-
hydroxysuccinimide, 1-hydroxybenzotriazole and the like, may be used to
facilitate the coupling reaction.
This coupling reaction is typically conducted by contacting the N
sulfonylamino acid IV with about 1 to about 2 equivalents of the coupling
reagent and at least one equivalent, preferably about 1 to about 1.2
equivalents, of amino acid derivative VI in an inert diluent, such as
dichloromethane, chloroform, acetonitrile, tetrahydrofuran, N,N
dimethylformamide and the like. Generally, this reaction is conducted at a
temperature ranging from about 0°C to about 37°C for about 12 to
about 24
hours. Upon completion of the reaction, the compound of formula I is
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recovered by conventional methods including neutralization, extraction,
precipitation, chromatography, filtration, and the like.
Alternatively, the N sulfonyl amino acid IV can be converted into an
acid halide and the acid halide coupled with amino acid derivative VI to
provide compounds of formula I. The acid halide of VI can be prepared by
contacting VI with an inorganic acid halide, such as thionyl chloride,
phosphorous trichloride, phosphorous tribromide or phosphorous
pentachloride, or preferably, with oxalyl chloride under conventional
conditions. Generally, this reaction is conducted using about 1 to 5 molar
equivalents of the inorganic acid halide or oxalyl chloride, either neat or in
an inert solvent, such as dichloromethane or carbon tetrachloride, at a
temperature in the range of about 0°C to about 80°C for about 1
to about 48
hours. A catalyst, such as N,N-dimethylformamide, may also be used in this
reaction.
The acid halide of N sulfonyl amino acid IV is then contacted with at
least one equivalent, preferably about 1.1 to about 1.5 equivalents, of amino
acid derivative VI in an inert diluent, such as dichloromethane, at a
temperature ranging from about -70°C to about 40°C for about 1
to about 24
hours. Preferably, this reaction is conducted in the presence of a suitable
base to scavenge the acid generated during the reaction. Suitable bases
include, by way of example, tertiary amines, such as triethylamine,
diisopropylethylamine, N methylmorpholine and the like. Alternatively, the
reaction can be conducted under Schotten-Baumann-type conditions using
aqueous alkali, such as sodium hydroxide and the like. Upon completion of
the reaction, the compound of formula I is recovered by conventional
methods including neutralization, extraction, precipitation. chromatography,
filtration, and the like.
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Alternatively, the compounds of formula I can be prepared by first
forming a diamino acid derivative of formula VII:
R3 R' O
II
RZ-NH-C-C(O)N-CH-C-R6 VII
H RS
wherein R2, R3, R5, R6, and R' are as defined above. The diamino acid
derivatives of formula VII can be readily prepared by coupling an amino acid
of formula II with an amino acid derivative of formula VI using conventional
amino acid coupling techniques and reagents, such carbodiimides, BOP
reagent and the like, as described above. Diamino acid VII can then be
sulfonated using a sulfonyl chloride of formula III and using the synthetic
procedures described above to provide a compound of formula I.
The amino acid derivatives of formula VI employed in the above
reactions are either known compounds or compounds that can be prepared
from known compounds by conventional synthetic procedures. For example,
amino acid derivatives of formula VI can be prepared by C-alkylating
commercially available diethyl 2-acetamidomalonate (Aldrich, Milwaukee,
Wisconsin, USA) with an alkyl or substituted alkyl halide. This reaction is
typically conducted by treating the diethyl 2-acetamidomalonate with at least
one equivalent of sodium ethoxide and at least one equivalent of an alkyl or
substituted alkyl halide in refluxing ethanol for about 6 to about 12 hours.
The resulting C-alkylated malonate is then deacetylated, hydrolyzed and
decarboxylated by heating in aqueous hydrochloric acid at reflux for about 6
to about 12 hours to provide the amino acid, typically as the hydrochloride
salt.
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Examples of amino acid derivatives of formula VI suitable for use in
the above reactions include, but are not limited to, L-alanine methyl ester, L-
isoleucine methyl ester, L-leucine methyl ester, L-valine methyl ester, ~3-
tert-
butyl-L-aspartic acid methyl ester, L-asparagine tert-butyl ester, E-Boc-L-
lysine methyl ester, E-Cbz-L-lysine methyl ester, y-tert-butyl-L-glutamic
acid methyl ester, L-glutamine tert-butyl ester, L-(N methyl)histidine methyl
ester, L-(N benzyi)histidine methyl ester, L-methionine methyl ester, L-(D-
benzyl)serine methyl ester, L-tryptophan methyl ester, L-phenylalanine
methyl ester, L-phenylalanine isopropyl ester, L-phenylalanine benzyl ester,
L-phenylalaninamide, N-methyl-L-phenylalanine benzyl ester, 3-carboxy-
D,L-phenylalanine methyl ester, 4-carboxy-D,L-phenylalanine methyl ester,
L-4-chiorophenylalanine methyl ester, L-4-(3-
dimethylaminopropyloxy)phenylalanine methyl ester, L-4-iodophenylalanine
methyl ester, L-3,4-methylenedioxyphenylalanine methyl ester, L-3,4-
ethylenedioxyphenylalanine methyl ester, L-4-nitrophenylalanine methyl
ester, L-tyrosine methyl ester, D,L-homophenylalanine methyl ester, L-(O-
methyl)tyrosine methyl ester, L-(D-tert-butyi)tyrosine methyl ester, L-(O-
benzyl)tyrosine methyl ester, L-3,5-diiodotyrosine methyl ester, L-3-
iodotyrosine methyl ester, (3-(1-naphthyl)-L-alanine methyl ester, (3-(2-
naphthyl)-L-alanine methyl ester, ~3-(2-thienyl)-L-alanine methyl ester, (3-
cyclohexyl-L-alanine methyl ester, ~3-(2-pyridyl)-L-alanine methyl ester, ~i-
(3-pyridyl)-L-aianine methyl ester, ~i-(4-pyridyl)-L-alanine methyl ester, ~i-
(2-thiazolyl)-D,L-alanine methyl ester, (3-(1,2,4-triazol-3-yl)-D,L-alanine
methyl ester, and the like. If desired, of course, other esters or amides of
the
above-described compounds may also be employed.
For ease of synthesis, the compounds of formula I are typically
prepared as an ester, i.e., where R6 is an alkoxy or substituted alkoxy group
and the like. If desired, the ester group can be hydrolysed using conventional
conditions and reagents to provide the corresponding carboxylic acid.
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Typically, this reaction is conducted by treating the ester with at least one
equivalent of an alkali metal hydroxide, such as lithium, sodium or
potassium hydroxide, in an inert diluent, such as methanol or mixtures of
methanol and water, at a temperature ranging about 0°C to about
24°C for
about 1 to about 12 hours. Alternatively, benzyl esters may be removed by
hydrogenolysis using a palladium catalyst, such as palladium on carbon. The
resulting carboxylic acids may be coupled, if desired, to amines such as
~i-alanine ethyl ester, hydroxyamines such as hydroxylamine and
N hydroxysuccinimide, alkoxyamines and substituted alkoxyamines such as
O-methylhydroxylamine and O-benzylhydroxylamine, and the like, using
conventional coupling reagents and conditions as described above.
As will be apparent to those skilled in the art, other functional groups
present on any of the substituents of the compounds of formula I can be
readily modified or derivatized either before or after the above-described
coupling reactions using well-known synthetic procedures. For example, a
nitro group present on a substituent of a compound of formula I or an
intermediate thereof may be readily reduced by hydrogenation in the
presence of a palladium catalyst, such as palladium on carbon, to provide the
corresponding amino group. This reaction is typically conducted at a
temperature of from about 20°C to about 50°C for about 6 to
about 24 hours
in an inert diluent, such as methanol. Compounds having a nitro group on
the RS substituent can be prepared, for example, by using a 4-
nitrophenylalanine derivative and the like in the above-described coupling
reactions.
Similarly, a pyridyl group can be hydrogenated in the presence of a
platinum catalyst, such as platinum oxide, in an acidic diluent to provide the
corresponding piperidinyl analogue. Generally, this reaction is conducted by
treating the pyridine compound with hydrogen at a pressure ranging from
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about 20 psi to about 60 psi, preferably about 40 psi, in the presence of the
catalyst at a temperature of about 20°C to about 50°C for about
2 to about 24
hours in an acidic diluent, such as a mixture of methanol and aqueous
hydrochloric acid. Compounds having a pyridyl group can be readily
prepared by using, for example, ~3-(2-pyridyl)-, ~i-(3-pyridyl)- or ~i-(4-
pyridyl)-L-alanine derivatives in the above-described coupling reactions.
Additionally, when the RS substituent of a compound of formula I or
an intermediate thereof contains a primary or secondary amino group, such
amino groups can be further derivatized either before or after the above
coupling reactions to provide, by way of example, amides, sulfonamides,
areas, thioureas, carbamates, secondary or tertiary amines and the like.
Compounds having a primary amino group on the RS substituent may be
prepared, for example, by reduction of the corresponding nitro compound as
described above. Alternatively, such compounds can be prepared by using
an amino acid derivative of formula VI derived from lysine,
4-aminophenylalanine and the like in the above-described coupling reactions.
By way of illustration, a compound of formula I or an intermediate
thereof having a substituent containing a primary or secondary amino group,
such as where RS is a (4-aminophenyl)rnethyl group, can be readily
N acylated using conventional acylating reagents and conditions to provide
the corresponding amide. This acylation reaction is typically conducted by
treating the amino compound with at least one equivalent, preferably about
1.1 to about 1.2 equivalents, of a carboxylic acid in the presence of a
coupling reagent such as a carbodiimide, BOP reagent (benzotriazol-1-yl-
oxy-tris{dimethylamino)phosphonium hexafluorophosphonate) and the like,
in an inert diluent, such as dichloromethane, chloroform, acetonitrile,
tetrahydrofuran, N,N dimethylformamide and the like, at a temperature
ranging from about 0°C to about 37°C for about 4 to about 24
hours.
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Preferably, a promoter, such as N hydroxysuccinimide, 1-hydroxy-
benzotriazole and the like, is used to facilitate the acylation reaction.
Examples of carboxylic acids suitable for use in this reaction include, but
are
not limited to, N tert-butyloxycarbonylglycine, N tent-butyloxycarbonyl-L-
phenylalanine, N tert-butyloxycarbonyl-L-aspartic acid benzyl ester, benzoic
acid, N tert-butyloxycarbonylisonipecotic acid, N methylisonipecotic acid,
N tert-butyloxycarbonylnipecotic acid, N tert-butyloxycarbonyl-L-
tetrahydroisoquinoline-3-carboxylic acid, N (toluene-4-sulfonyl)-L-proline
and the like.
Alternatively, a compound of formula I or an intermediate thereof
containing a primary or secondary amino group can be N acylated using an
acyl halide or a carboxylic acid anhydride to form the corresponding amide.
This reaction is typically conducted by contacting the amino compound with
at least one equivalent, preferably about 1.1 to about 1.2 equivalents, of the
acyl halide or carboxylic acid anhydride in an inert diluent, such as
dichloromethane, at a temperature ranging from about of about -70°C to
about 40°C for about 1 to about 24 hours. If desired, an acylation
catalyst
such as 4-(N,N dimethylamino)pyridine may be used to promote the
acylation reaction. The acylation reaction is preferably conducted in the
presence of a suitable base to scavenge the acid generated during the
reaction. Suitable bases include, by way of example, tertiary amines, such as
triethylamine, diisopropylethylamine, N-methylmorpholine and the like.
Alternatively, the reaction can be conducted under Schotten-Baumann-type
conditions using aqueous alkali, such as sodium hydroxide and the like.
Examples of acyl halides and carboxylic acid anhydrides suitable for
use in this reaction include, but are not limited to, 2-methylpropionyl
chloride, trimethylacetyl chloride, phenylacetyl chloride, benzoyl chloride, 2-
bromobenzoyl chloride, 2-methylbenzoyl chloride, 2-trifluoro-
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methylbenzoyl chloride, isonicotinoyl chloride, nicotinoyl chloride,
picolinoyl chloride, acetic anhydride, succinic anhydride, and the like.
Carbamyl chlorides, such as N,N dimethylcarbamyl chloride, N,N
diethylcarbamyl chloride and the like, can also be used in this reaction to
provide areas. Similarly, dicarbonates, such as di-tert-butyl dicarbonate,
may be employed to provide carbamates.
In a similar manner, a compound of formula I or an intermediate
thereof containing a primary or secondary amino group may be N sulfonated
to form a sulfonamide using a sulfonyl halide or a sulfanic acid anhydride.
Sulfonyl halides and sulfonic acid anhydrides suitable for use in this
reaction
include, but are not limited to, methanesulfonyl chloride,
chloromethanesulfonyl chloride, p-toluenesulfonyl chloride,
trifluoromethanesulfonic anhydride, and the like. Similarly, sulfamoyl
chlorides, such as dimethylsulfamoyl chloride, can be used to provide
sulfamides (e.g., >N-SO~-N<).
Additionally, a primary and secondary amino group present on a
substituent of a compound of formula I or an intermediate thereof can be
reacted with an isocyanate or a thioisocyanate to give a urea or thiourea,
respectively. This reaction is typically conducted by contacting the amino
compound with at least one equivalent, preferably about 1.1 to about 1.2
equivalents, of the isocyanate or thioisocyanate in an inert diluent, such as
toluene and the like, at a temperature ranging from about 24 °C to
about
37°C for about 12 to about 24 hours. The isocyanates and
thioisocyanates
used in this reaction are commercially available or can be prepared from
commercially available compounds using well-known synthetic procedures.
For example, isocyanates and thioisocyanates are readily prepared by
reacting the appropriate amine with phosgene or thiophosgene. Examples of
isocyanates and thioisocyanates suitable for use in this reaction include, but
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are not limited to, ethyl isocyanate, n-propyl isocyanate, 4-cyanophenyl
isocyanate, 3-methoxyphenyl isocyanate, 2-phenylethyl isocyanate, methyl
thioisocyanate, ethyl thioisocyanate, 2-phenylethyl thioisocyanate, 3-
phenylpropyl thioisocyanate, 3-(N,N diethylamino)propyl thioisocyanate,
phenyl thioisocyanate, benzyl thioisocyanate, 3-pyridyl thioisocyanate,
fluorescein isothiocyanate (isomer I) and the like.
Furthermore, when a compound of formula I or IA, or an
intermediate thereof, contains a primary or secondary amino group, the
amino group can be reductively alkylated using aldehydes or ketones to form
a secondary or tertiary amino group. This reaction is typically conducted by
contacting the amino compound with at least one equivalent, preferably
about 1.1 to about 1.5 equivalents, of an aldehyde or ketone and at least one
equivalent based on the amino compound of a metal hydride reducing agent,
such as sodium cyanobarohydride, in an inert diluent, such as methanol,
tetrahydrofuran, mixtures thereof and the like, at a temperature ranging from
about 0°C to about SO°C for about 1 to about 72 hours. Aldehydes
and
ketones suitable for use in this reaction include, by way of example,
benzaldehyde, 4-chlorobenzaldehyde, valeraldehyde and the like.
In a similar manner, when a compound of formula I or an
intermediate thereof has a substituent containing a hydroxyl group, the
hydroxyl group can be further modified or derivatized either before or after
the above coupling reactions to provide, by way of example, ethers,
carbamates and the like. Compounds having a hydroxyl group on the RS
substituent, for example, can be prepared using an amino acid derivative of
formula VI derived from tyrosine and the like in the above-described
reactions.
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By way of example, a compound of formula I or an intermediate
thereof having a substituent containing a hydroxyl group, such as where Rs is
a (4-hydroxyphenyl)methyl group, can be readily O-alkylated to form ethers.
This O-alkylation reaction is typically conducted by contacting the hydroxy
compound with a. suitable alkali or alkaline earth metal base, such as
potassium carbonate, in an inert diluent, such as acetone, 2-butanone and the
like, to form the alkali or alkaline earth metal salt of the hydroxyl group.
This salt is generally not isolated, but is reacted in situ with at least one
equivalent of an alkyl or substituted alkyl halide or sulfonate, such as an
alkyl chloride, bromide, iodide, mesylate or tosylate, to afford the ether.
Generally, this reaction is conducted at a temperature ranging from about
60 ° C to about 150 ° C for about 24 to about 72 hours.
Preferably, a catalytic
amount of sodium or potassium iodide is added to the reaction mixture when
an alkyl chloride or bromide is employed in the reaction.
Examples of alkyl or substituted alkyl halides and sulfonates suitable
for use in this reaction include, but are not limited to, tent-butyl
bromoacetate, N tent-butyl chloroacetamide, 1-bromoethylbenzene, ethyl a-
bromophenylacetate, 2-(N ethyl-N phenylamino)ethyl chloride, 2-(N,N-
ethylamino)ethyl chloride, 2-{N,IV diisopropylamino)ethyl chloride, 2-(N,N
dibenzylamino)ethyl chloride, 3-(N,N ethylamino)propyl chloride, 3-(N
benzyl-N methylamino)propyl chloride, N-(2-chloroethyl)morpholine, 2-
(hexamethyleneimino)ethyl chloride, 3-(N-methylpiperazine)propyl chloride,
1-(3-chlorophenyl)-4-(3-chloropropyl)piperazine, 2-(4-hydroxy-4-
phenylpiperidine)ethyl chloride, N tent-butyloxycarbonyl-3-piperidinemethyl
tosylate, and the like.
Alternatively, a hydroxyl group present on a substituent of a
compound of formula I or an intermediate thereof can be O-alkylating using
the Mitsunobu reaction. In this reaction, an alcohol, such as 3-(N,N
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dimethylamino)-1-propanol and the like, is reacted with about 1.0 to about
1.3 equivalents of triphenylphosphine and about 1.0 to about 1.3 equivalents
of diethyl azodicarboxylate in an inert diluent, such as tetrahydrofuran, at a
temperature ranging from about -10 ° C to about 5 ° C for about
0.25 to about
1 hour. About 1.0 to about 1.3 equivalents of a hydroxy compound, such as
N tert-butyltyrosine methyl ester, is then added and the reaction mixture is
stirred at a temperature of about 0°C to about 30°C for about 2
to about 48
hours to provide the O-alkylated product.
In a similar manner, a compound of formula I or an intermediate
thereof containing a aryl hydroxy group can be reacted with an aryl iodide to
provide a diaryl ether. Generally, this reaction is conducted by forming the
alkali metal salt of the hydroxyl group using a suitable base, such as sodium
hydride, in an inert diluent such as xylenes at a temperature of about -25
°C
to about 10 ° C. The salt is then treated with about 1.1 to about 1.5
equivalents of cuprous bromide dimethyl sulfide complex at a temperature
ranging from about 10 ° C to about 30 ° C for about 0.5 to about
2.0 hours,
followed by about 1.1 to about 1.5 equivalents of an aryl iodide, such as
sodium 2-iodobenzoate and the like. The reaction is then heated to about
70°C to about 150°C for about 2 to about 24 hours to provide the
diaryl
ether.
Additionally, a hydroxy-containing compound can also be readily
derivatized to form a carbamate. In one method for preparing such
carbamates, a hydroxy compound of formula I or an intermediate thereof is
contacted with about 1.0 to about 1.2 equivalents of 4-nitrophenyl
chloroformate in an inert diluent, such as dichloromethane, at a temperature
ranging from about -25°C to about 0°C for about 0.5 to about 2.0
hours.
Treatment of the resulting carbonate with an excess, preferably about 2 to
about 5 equivalents, of a trialkylamine, such as triethylamine, for about 0.5
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to 2 hours, followed by about 1.0 to about 1.5 equivalents of a primary or
secondary amine provides the carbamate. Examples of amines suitable for
using in this reaction include, but are not limited to, piperazine, 1-
methylpiperazine, 1-acetylpiperazine, morpholine, thiomorpholine,
pyrrolidine, piperidine and the like.
Alternatively, in another method for preparing carbamates, a
hydroxy-containing compound is contacted with about 1.0 to about 1.5
equivalents of a carbamyl chloride in an inert diluent, such as
dichloromethane, at a temperature ranging from about 25 °C to about
70°C
for about 2 to about 72 hours. Typically, this reaction is conducted in the
presence of a suitable base to scavenge the acid generated during the
reaction. Suitable bases include, by way of example, tertiary amines, such as
triethylamine, diisopropylethylamine, N methylmorpholine and the like.
Additionally, at least one equivalent (based on the hydroxy compound) of 4-
(N,N dimethylamino)pyridine is preferably added to the reaction mixture to
facilitate the reaction. Examples of carbamyl chlorides suitable for use in
this reaction include, by way of example, dimethylcarbamyl chloride,
diethylcarbamyl chloride and the like.
Likewise, when a compound of formula I or an intermediate thereof
contains a primary or secondary hydroxyl group, such hydroxyl groups can
be readily converted into a leaving group and displaced to form, for example,
amines, sulfides and fluorides. For example, derivatives of 4-hydroxy-L-
proline can be converted into the corresponding 4-amino, 4-thin or 4-fluoro-
L-proline derivatives via nucleophilic displacement of the derivatized
hydroxyl group. Generally, when a chiral compound is employed in these
reactions, the stereochemistry at the carbon atom attached to the derivatized
hydroxyl group is inverted.
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These reactions are typically conducted by first converting the
hydroxyl group into a leaving group, such as a tosylate, by treatment of the
hydroxy compound with at least one equivalent of a sulfonyl halide, such as
p-toluenesulfonyl chloride and the like, in pyridine. This reaction is
generally conducted at a temperature of from about 0°C to about
70°C for
about 1 to about 48 hours. The resulting tosylate can then be readily
displaced with sodium azide, for example, by contacting the tosylate with at
least one equivalent of sodium azide in an inert diluent, such as a mixture of
N,N dimethylformamide and water, at a temperature ranging from about
0°C
to about 37°C for about 1 to about 12 hours to provide the
corresponding
azido compound. The azido group can then be reduced by, for example,
hydrogenation using a palladium on carbon catalyst to provide the amino (-
NH2) compound.
1 S Similarly, a tosylate group can be readily displaced by a thiol to form
a sulfide. This reaction is typically conducted by contacting the tosylate
with
at least one equivalent of a thiol, such as thiophenol, in the presence of a
suitable base, such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), in an inert
diluent, such as N,N dimethylformamide, at a temperature of from about
0°C
to about 37°C for about 1 to about 12 hours to provide the sulfide.
Additionally, treatment of a tosylate with morpholinosulfur trifluoride in an
inert diluent, such as dichloromethane, at a temperature ranging from about
0°C to about 37°C for about 12 to about 24 hours affords the
corresponding
fluoro compound.
Furthermore, a compound of formula I or IA or an intermediate
thereof having a substituent containing an iodoaryl group, for example, when
RS is a (4-iodophenyl)methyl group, can be readily converted either before or
after the above coupling reactions into a biaryl compound. Typically, this
reaction is conducted by treating the iodoaryl compound with about 1.1 to
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about 2 equivalents of an arylzinc iodide, such as 2-(methoxycarbonyl)-
phenylzinc iodide, in the presence of a palladium catalyst, such as palladium
tetra(triphenylphosphine), in an inert diluent, such as tetrahydrofuran, at a
temperature ranging from about 24°C to about 30°C until reaction
completion. This reaction is further described, for example, in Rieke, J. Org.
Chem. 1991, 56, 1445.
In some cases, the compounds of formula I or IA or intermediates
thereof may contain substituents having one or more sulfur atoms. Such
sulfur atoms will be present, for example, when the amino acid of formula II
employed in the above reactions is derived from L-thiazolidine-4-carboxylic
acid, L-(5,5-dimethyl)thiazolidine-4-carboxylic acid, L-thiamorpholine-3-
carboxylic acid and the like. When present, such sulfur atoms can be
oxidized either before or after the above coupling reactions to provide a
sulfoxide or sulfone compound using conventional reagents and reaction
conditions. Suitable reagents for oxidizing a sulfide compound to a
sulfoxide include, by way of example, hydrogen peroxide,
3-chloroperoxybenzoic acid (MCPBA), sodium periodate and the like. The
oxidation reaction is typically conducted by contacting the sulfide compound
with about 0.95 to about 1.1 equivalents of the oxidizing reagent in an inert
diluent, such as dichloromethane, at a temperature ranging from about -
50°C
to about 75 °C for about 1 to about 24 hours. The resulting sulfoxide
can
then be further oxidized to the corresponding sulfone by contacting the
sulfoxide with at least one additional equivalent of an oxidizing reagent,
such
as hydrogen peroxide, MCPBA, potassium permanganate and the like.
Alternatively, the sulfone can be prepared directly by contacting the sulfide
with at least two equivalents, and preferably an excess, of the oxidizing
reagent. Such reactions are described further in March, "Advanced Organic
Chemistry", 4th Ed., pp. 1201-1202, Wiley Publisher, 1992.
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As described above, the compounds of formula I having an RZ
substituent other an hydrogen can be prepared using an N substituted amino
acid of formula II, such as sarcosine, N methyl-L-phenylalanine and the like,
in the above-described coupling reactions. Alternatively, such compounds
can be prepared by N alkylation of a sulfonamide of formula I or IV (where
RZ is hydrogen) using conventional synthetic procedures. Typically, this N
alkylation reaction is conducted by contacting the sulfonamide with at least
one equivalent, preferably 1.1 to 2 equivalents, of an alkyl or substituted
alkyl halide in the presence of a suitable base, such as potassium carbonate,
in an inert diluent, such as acetone, 2-butanone and the like, at a
temperature
ranging from about 25°C to about 70°C for about 2 to about 48
hours.
Examples of alkyl or substituted alkyl halides suitable for use in this
reaction
include, but are not limited to, methyl iodide, and the like.
Additionally, the sulfonamides of formula I or IV wherein R' is
hydrogen and R' is a 2-alkoxycarbonylaryl group can be intramolecularly
cyclized to form 1,2-benzisothiazol-3-one derivatives or analogues thereof.
This reaction is typically conducted by treating a sulfonamide, such as N (2-
methoxycarbonylphenylsulfonyl)glycine-L-phenylalanine benzyl ester, with
about 1.0 to 1.5 equivalents of a suitable base, such as an alkali metal
hydride, in a inert diluent, such as tetrahydrofuran, at a temperature ranging
from about 0°C to about 30°C for about 2 to about 48 hours to
afford the
cyclized 1,2-benzisothiazol-3-one derivative.
Lastly, the compounds of formula I where Q is -C(S)NR'- can be
prepared by using an amino thionoacid derivative in place of amino acid II in
the above described synthetic procedures. Such amino thionoacid derivatives
can be prepared by the procedures described in Shalaky, et al., J. Org.
Chem., øx:9045-9048 (1996) and Brain, et al., J. Org. Chem., øx:3808-3809
( 1997) and references cited therein.
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Pharmaceutical Formulations
When employed as pharmaceuticals, the compounds of formula I and
IA are usually administered in the form of pharmaceutical compositions.
These compounds can be administered by a variety of routes including oral,
rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal.
These compounds are effective as both injectable and oral compositions.
Such compositions are prepared in a manner well known in the
pharmaceutical art and include at least one active compound.
This invention alsa includes pharmaceutical compositions which
contain, as the active ingredient, one or more of the compounds of formula I
and IA above associated with pharmaceutically acceptable Garners. In
making the compositions of this invention, the active ingredient is usually
mixed with an excipient, diluted by an excipient or enclosed within such a
carrier which can be in the form of a capsule, sachet, paper or other
container. When the excipient serves as a diluent, it can be a solid, semi-
solid, or liquid material, which acts as a vehicle, carrier or medium for the
active ingredient. Thus, the compositions can be in the form of tablets,
pills,
powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions,
solutions, syrups, aerosols (as a solid or in a liquid medium), ointments
containing, for example, up to 10% by weight of the active compound, soft
and hard gelatin capsules, suppositories, sterile injectable solutions, and
sterile packaged powders.
In preparing a formulation, it may be necessary to mill the active
compound to provide the appropriate particle size prior to combining with
the other ingredients. If the active compound is substantially insoluble, it
ordinarily is milled to a particle size of less than 200 mesh. If the active
compound is substantially water soluble, the particle size is normally
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adjusted by milling to provide a substantially uniform distribution in the
formulation, e.g. about 40 mesh.
Some examples of suitable excipients include lactose, dextrose,
sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate,
alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose,
polyvinylpyrrolidone, cellulose, water, syrup, and methyl cellulose. The
formulations can additionally include: lubricating agents such as talc,
magnesium stearate, and mineral oil; wetting agents; emulsifying and
suspending agents; preserving agents such as methyl- and propylhydroxy-
benzoates; sweetening agents; and flavoring agents. The compositions of the
invention can be formulated so as to provide quick, sustained or delayed
release of the active ingredient after administration to the patient by
employing procedures known in the art.
The compositions are preferably formulated in a unit dosage form,
each dosage containing from about 5 to about 100 mg. more usually about 10
to about 30 mg, of the active ingredient. The term "unit dosage forms" refers
to physically discrete units suitable as unitary dosages for human subjects
and other mammals, each unit containing a predetermined quantity of active
material calculated to produce the desired therapeutic effect, in association
with a suitable pharmaceutical excipient.
The active compound is effective over a wide dosage range and is
generally administered in a pharmaceutically effective amount. It, will be
understood, however, that the amount of the compound actually administered
will be determined by a physician, in the light of the relevant circumstances,
including the condition to be treated, the chosen route of administration, the
actual compound administered, the age, weight, and response of the
individual patient, the severity of the patient's symptoms, and the like.
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For preparing solid compositions such as tablets, the principal active
ingredient is mixed with a pharmaceutical excipient to form a solid
preformulation composition containing a homogeneous mixture of a
compound of the present invention. When referring to these preformulation
compositions as homogeneous, it is meant that the active ingredient is
dispersed evenly throughout the composition so that the composition may be
readily subdivided into equally effective unit dosage forms such as tablets,
pills and capsules. This solid preformulation is then subdivided into unit
dosage forms of the type described above containing from, for example, 0.1
to about 500 mg of the active ingredient of the present invention.
The tablets or pills of the present invention may be coated or
otherwise compounded to provide a dosage form affording the advantage of
prolonged action. For example, the tablet or pill can include an inner dosage
and an outer dosage component, the latter being in the form of an envelope
over the former. The two components can separated by an enteric layer
which serves to resist disintegration in the stomach and permit the inner
component to pass intact into the duodenum or to be delayed in release. A
variety of materials can be used for such enteric layers or coatings, such
materials including a number of polymeric acids and mixtures of polymeric
acids with such materials as shellac, cetyl alcohol, and cellulose acetate.
The liquid forms in which the compositions of the present invention
may be incorporated for administration orally or by injection include
aqueous solutions suitably flavored syrups, aqueous or oil suspensions, and
flavored emulsions with edible oils such as cottonseed oil, sesame oil,
coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical
vehicles.
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Compositions for inhalation or insufflation include solutions and
suspensions in pharmaceutically acceptable, aqueous or organic solvents, or
mixtures thereof, and powders. The liquid or solid compositions may
contain suitable pharmaceutically acceptable excipients as described supra.
Preferably the compositions are administered by the oral or nasal respiratory
route for local or systemic effect. Compositions in preferably
pharmaceutically acceptable solvents may be nebulized by use of inert gases.
Nebulized solutions may be breathed directly from the nebulizing device or
the nebulizing device may be attached to a face mask tent, or intermittent
positive pressure breathing machine. Solution, suspension, or powder
compositions may be administered, preferably orally or nasally, from devices
which deliver the formulation in an appropriate manner.
The following formulation examples illustrate the pharmaceutical
compositions of the present invention.
Formulation Example 1
Hard gelatin capsules containing the following ingredients are
prepared:
Quantity
Ingredient e/capsulel
Active Ingredient 30.0
Starch 305.0
Magnesium stearate 5.0
The above ingredients are mixed and filled into hard gelatin capsules
in 340 mg quantities.
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Formulation Example 2
A tablet formula is prepared using the ingredients below:
Quantity
Active Ingredient 25.0
Cellulose, microcrystalline 200.0
Colloidal silicon dioxide 10.0
Stearic acid 5.0
The components are blended and compressed to form tablets, each
weighing 240 mg.
Formulation Example 3
A dry powder inhaler formulation is prepared containing the
following components:
n~Wei t
Active Ingredient 5
Lactose 95
The active mixture is mixed with the lactose and the mixture is added
to a dry powder inhaling appliance.
Formulation Example 4
Tablets, each containing 30 mg of active ingredient, are prepared as
follows:
Quantity
Ir~;redient ~n /ta et
Active Ingredient 30.0 mg
Starch 45.0 mg
Microcrystalline cellulose 35.0 mg
Polyvinylpyrrolidone
(as 10% solution in water) 4.0 mg
Sodium carboxymethyl starch 4.5 mg
Magnesium stearate 0.5 mg
Talc 1.0 mg
Total 120 mg
*rB
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The active ingredient, starch and cellulose are passed through a No.
20 mesh U.S. sieve and mixed thoroughly. The solution of polyvinyl-
pyrrolidone is mixed with the resultant powders, which are then passed
through a 16 mesh U.S. sieve. The granules so produced are dried at 50°
to
60°C and passed through a 16 mesh U.S. sieve. The sodium carboxymethyl
starch, magnesium stearate, and talc, previously passed through a No. 30
mesh U.S. sieve, are then added to the granules which, after mixing, are
compressed on a tablet machine to yield tablets each weighing 150 mg.
Formulation Example 55
Capsules, each containing 40 mg of medicament are made as follows:
Quantity
InQredien~ fmg/c_a sp ulel_
Active Ingredient 40.0 mg
Starch 109.0 mg
Magnesium stearate 1.0 m~
Total 150.0 mg
The active ingredient, cellulose, starch, an magnesium stearate are
blended, passed through a No. 20 mesh U.S. sieve, and filled into hard
gelatin capsules in 150 mg quantities.
Formulation Exnle 6
Suppositories, each containing 25 mg of active ingredient are made as
follows:
In ' t Amount
Active Ingredient 25 mg
Saturated fatty acid glycerides to 2,000 mg
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The active ingredient is passed through a No. 60 mesh U.S. sieve and
suspended in the saturated fatty acid glycerides previously melted using the
minimum heat necessary. The mixture is then poured into a suppository
mold of nominal 2.0 g capacity and allowed to cool.
Forr,~,ulat~~ E~ xam~e 7
Suspensions, each containing 50 mg of medicament per 5.0 ml dose
are made as follows:
Ingredi~t Amount
Active Ingredient SO.OOmg
Xanthan gum 4.OOmg
Sodium carboxymethyl cellulose ( 11 %)
Microcrystalline cellulose (89%) SO.OOmg
Sucrose 1.75 g
Sodium benzoate 10.00mg
Flavor and Color q.v.
Purified water to S.OOm1
The medicament, sucrose and xanthan gum are blended, passed
through a No. 10 mesh U.S. sieve, and then mixed with a previously made
solution of the microcrystalline cellulose and sodium carboxymethyl
cellulose in water. The sodium benzoate, flavor, and color are diluted with
some of the water and added with stirring. Sufficient water is then added to
produce the required volume.
Formulation l~xamp~e 8
Quantity
~gredient (,m~/ca sh ulel_
Active Ingredient 15.0 mg
Starch 407.0 mg
Magnesium stearate 3.0 me
Total 425.0 mg
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The active ingredient, cellulose, starch, and magnesium stearate are
blended, passed through a No. 20 mesh U.S. sieve, and filled into hard
gelatin capsules in 560 mg quantities.
Formulation Example 9
An intravenous formulation may be prepared as follows:
In re ' t Quantitv
Active Ingredient 250.0 mg
Isotonic saline 1000.0 ml
Formulation Examnl_~ 10
A topical formulation may be prepared as follows:
1 S I ~ ' t a tit
Active Ingredient 1-10 g
Emulsifying Wax 30 g
Liquid Paraffin 20 g
White Soft Paraffin to 100 g
The white soft paraffin is heated until molten. The liquid paraffin
and emulsifying wax are incorporated and stirred until dissolved. The active
ingredient is added and stirring is continued until dispersed. The mixture is
then cooled until solid.
Another preferred formulation employed in the methods of the
present invention employs transdermal delivery devices ("patches"). Such
transdermal patches may be used to provide continuous or discontinuous
infusion of the compounds of the present invention in controlled amounts.
The construction and use of transdermal patches for the delivery of
pharmaceutical agents is well known in the art. ~~g. e~,~., U.S. Patent
5,023,252, issued June 11, 1991, herein incorporated by reference. Such
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patches may be constructed for continuous, pulsatile, or on demand delivery
of pharmaceutical agents.
It may be desirable or necessary to introduce the pharmaceutical
composition to the brain, either directly or indirectly. Direct techniques
usually involve placement of a drug delivery catheter into the host's
ventricular system to bypass the blood-brain barrier. One such implantable
delivery system used for the transport of biological factors to specific
anatomical regions of the body is described in U.S. Patent 5,011,472 which
is herein incorporated by reference.
Indirect techniques, which are generally preferred, usually involve
formulating the compositions to provide for drug latentiation by the
conversion of hydrophilic drugs into lipid-soluble drugs. Latentiation is
generally achieved through blocking of the hydroxy, carbonyl, sulfate, and
primary amine groups present on the drug to render the drug more lipid
soluble and amenable to transportation across the blood-brain barrier.
Alternatively, the delivery of hydrophilic drugs may be enhanced by
intra-arterial infusion of hypertonic solutions which can transiently open the
blood-brain barner.
tilt
The compounds of this invention can be employed to bind VLA-4
(a4(3, integrin) in biological samples and, accordingly have utility in, for
example, assaying such samples for VLA-4. In such assays, the compounds
can be bound to a solid support and the VLA-4 sample added thereto. The
amount of VLA-4 in the sample can be determined by conventional methods
such as use of a sandwich ELISA assay. Alternatively, labeled VLA-4 can
be used in a competitive assay to measure for the presence of VLA-4 in the
sample. Other suitable assays are well known in the art.
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In addition, certain of the compounds of this invention inhibit, in
vivo, adhesion of leukocytes to endothelial cells mediated by VLA-4 and,
accordingly, can be used in the treatment of diseases mediated by VLA-4.
Such diseases include inflammatory diseases in mammalian patients
such as asthma, Alzheimer's disease, atherosclerosis, AIDS dementia,
diabetes (including acute juvenile onset diabetes), inflammatory bowel
disease (including ulcerative colitis and Crohn's disease), multiple
sclerosis,
rheumatoid arthritis, tissue transplantation, tumor metastasis, meningitis,
encephalitis, stroke, and other cerebral traumas, nephritis, retinitis, atopic
dermatitis, psoriasis, myocardial ischemia and acute leukocyte-mediated lung
injury such as that which occurs in adult respiratory distress syndrome.
The biological activity of the compounds identified above may be
assayed in a variety of systems. For example, a compound can be
immobilized on a solid surface and adhesion of cells expressing VLA-4 can
be measured. Using such formats, large numbers of compounds can be
screened. Cells suitable for this assay include any leukocytes known to
express VLA-4 such as T cells, B cells, monocytes, eosinophils, and
basophils. A number of leukocyte cell lines can also be used, examples
include Jurkat and U937.
The test compounds can also be tested for the ability to competitively
inhibit binding between VLA-4 and VCAM-1, or between VLA-4 and a
labeled compound known to bind VLA-4 such as a compound of this
invention or antibodies to VLA-4. In these assays, the VCAM-1 can be
immobilized on a solid surface. VCAM-1 may also be expressed as a
recombinant fusion protein having an Ig tail (e.g., IgG) so that binding to
VLA-4 may be detected in an immunoassay. Alternatively, VCAM-19
expressing cells, such as activated endothelial cells or VCAM-1 transfected
fibroblasts can be used. For assays to measure the ability to block adhesion
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to brain endothelial cells, the assays described in International Patent
Application Publication No. WO 91/05038 are particularly preferred. This
application is incorporated herein by reference in its entirety.
Many assay formats employ labelled assay components. The
labelling systems can be in a variety of forms. The label may be coupled
directly or indirectly to the desired component of the assay according to
methods well known in the art. A wide variety of labels may be used. The
component may be labelled by any one of several methods. The most
common method of detection is the use of autoradiography with 3H, 'zSI, 3sS,
'4C, or 3'-P labelled compounds and the like. Non-radioactive labels include
ligands which bind to labelled antibodies, fluorophores, chemiluminescent
agents, enzymes and antibodies which can serve as specific binding pair
members for a labelled ligand. The choice of label depends on sensitivity
required, ease of conjugation with the compound, stability requirements, and
available instrumentation.
Appropriate in vivo models for demonstrating efficacy in treating
inflammatory responses include EAE (experimental autoimmune
encephalomyelitis) in mice, rats, guinea pigs or primates, as well as other
inflammatory models dependent upon a4 integrins.
Compounds having the desired biological activity may be modified
as necessary to provide desired properties such as improved pharmacological
properties (e.g., in vivo stability, bio-availability), or the ability to be
detected in diagnostic applications. For instance, inclusion of one or more
D-amino acids in the sulfonamides of this invention typically increases in
vivo stability. Stability can be assayed in a variety of ways such as by
measuring the half life of the proteins during incubation with peptidases or
human plasma or serum. A number of such protein stability assays have
*rB
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been described (see, e.g., Verhoef et aL, Eur. J. Drug Metab.
Pharmacokinet., 1990, ,1~2 :83-93).
For diagnostic purposes, a wide variety of labels may be linked to the
compounds, which may provide, directly or indirectly, a detectable signal.
Thus, the compounds of the subject invention may be modified in a variety
of ways for a variety of end purposes while still retaining biological
activity.
In addition, various reactive sites may be introduced at the terminus for
linking to particles, solid substrates, macromolecules, and the like.
IO
Labeled compounds can be used in a variety of in vivo or in vitro
applications. A wide variety of labels may be employed, such as
radionuclides (e.g., gamma-emitting radioisotopes such as technetium-99 or
indium-11 I), fluorescers (e.g., fluorescein), enzymes, enzyme substrates,
enzyme cofactors, enzyme inhibitors, chemiluminescent compounds,
bioluminescent compounds, and the like. Those of ordinary skill in the art
will know of other suitable labels for binding to the complexes, or will be
able to ascertain such using routine experimentation. The binding of these
labels is achieved using standard techniques common to those of ordinary
skill in the art.
In vitro uses include diagnostic applications such as monitoring
inflammatory responses by detecting the presence of leukocytes expressing
VLA-4. The compounds of this invention can also be used for isolating or
labeling such cells. In addition, as mentioned above, the compounds of the
invention can be used to assay for potential inhibitors of VLA-4NCAM-1
interactions.
For in vivo diagnostic imaging to identify, e.g., sites of inflammation,
radioisotopes are typically used in accordance with well known techniques.
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The radioisotopes may be bound to the peptide either directly or indirectly
using intermediate functional groups. For instance, chelating agents such as
diethylenetriaminepentacetic acid (DTPA) and ethylenediaminetetraacetic
acid (EDTA) and similar molecules have been used to bind proteins to
metallic ion radioisotopes.
The complexes can also be labeled with a paramagnetic isotope for
purposes of in vivo diagnosis, as in magnetic resonance imaging (MRI) or
electron spin resonance (ESR), both of which are well known. In general,
any conventional method for visualizing diagnostic images can be used.
Usually gamma- and positron-emitting radioisotopes are used for camera
imaging and paramagnetic isotopes are used for MRI. Thus, the compounds
can be used to monitor the course of amelioration of an inflammatory
response in an individual. By measuring the increase or decrease in
lymphocytes expressing VLA-4 it is possible to determine whether a
particular therapeutic regimen aimed at ameliorating the disease is effective.
The pharmaceutical compositions of the present invention can be
used to block or inhibit cellular adhesion associated with a number of
diseases and disorders, or to treat diseases in a mammalian patient which are
mediated by a9~i,. For instance, a number of inflammatory disorders are
associated with integrins or leukocytes. Treatable disorders include, e.g.,
transplantation rejection (e.g., allograft rejection), Alzheimer's disease,
atherosclerosis, AIDS dementia, diabetes (including acute juvenile onset
diabetes), retinitis, cancer metastases, rheumatoid arthritis, acute
leukocyte-mediated lung injury (e.g., adult respiratory distress syndrome),
asthma, nephritis, and acute and chronic inflammation, including atopic
dermatitis, psoriasis, myocardial ischemia, and inflammatory bowel disease
(including Crohn's disease and ulcerative colitis). In preferred embodiments
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the pharmaceutical compositions are used to treat inflammatory brain
disorders, such as multiple sclerosis (MS), viral meningitis and encephalitis.
Inflammatory bowel disease is a collective term for two similar
diseases referred to as Crohn's disease and ulcerative colitis. Crohn's
disease
is an idiopathic, chronic ulceroconstrictive inflammatory disease
characterized by sharply delimited and typically transmural involvement of
all layers of the bowel wall by a granulomatous inflammatory reaction. Any
segment of the gastrointestinal tract, from the mouth to the anus, may be
involved, although the disease most commonly affects the terminal ileum
and/or colon. Ulcerative colitis is an inflammatory response limited largely
to the colonic mucosa and submucosa. Lymphocytes and macrophages are
numerous in lesions of inflammatory bowel disease and may contribute to
inflammatory injury.
Asthma is a disease characterized by increased responsiveness of the
tracheobronchial tree to various stimuli potentiating paroxysmal constriction
of the bronchial airways. The stimuli cause release of various mediators of
inflammation from IgE-coated mast cells including histamine, eosinophilic
and neutrophilic chemotactic factors, leukotrines, prostaglandin and platelet
activating factor. Release of these factors recruits basophils, eosinophils
and
neutrophils, which cause inflammatory injury.
Atherosclerosis is a disease of arteries (e.g., coronary, carotid, aorta
and iliac). The basic lesion, the atheroma, consists of a raised focal plaque
within the intima, having a core of lipid and a covering fibrous cap.
Atheromas compromise arterial blood flow and weaken affected arteries.
Myocardial and cerebral infarcts are a major consequence of this disease.
Macrophages and leukocytes are recruited to atheromas and contribute to
inflammatory injury.
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Rheumatoid arthritis is a chronic, relapsing inflammatory disease that
primarily causes impairment and destruction of joints. Rheumatoid arthritis
usually first affects the small joints of the hands and feet but then may
involve the wrists, elbows, ankles and knees. T'he arthritis results from
interaction of synovial cells with leukocytes that infiltrate from the
circulation into the synovial lining of the joints. See e.g., Paul, Immunology
(3d ed., Raven Press, 1993).
Another indication for the compounds of this invention is in
treatment of organ or graft rejection mediated by VLA-4. Over recent years
there has been a considerable improvement in the efficiency of surgical
techniques for transplanting tissues and organs such as skin, kidney, liver,
heart, lung, pancreas and bone marrow. Perhaps the principal outstanding
problem is the lack of satisfactory agents for inducing immunotolerance in
the recipient to the transplanted allograft or organ. When allogenetic cells
or
organs are transplanted into a host (i.e., the donor and donee are different
individuals from the same species), the host immune system is likely to
mount an immune response to foreign antigens in the transplant (host-versus-
graft disease) leading to destruction of the transplanted tissue. CD8+ cells,
CD4 cells and monocytes are all involved in the rejection of transplant
tissues. Compounds of this invention which bind to alpha-4 integrin are
useful, inter alia, to block alloantigen-induced immune responses in the
donee thereby preventing such cells from participating in the destruction of
the transplanted tissue or organ. See, e.g., Paul et al., Transplant
International 9, 420-425 (1996); Georczynski et al., Immunology 87, 573-
580 (1996); Georcyznski et al., Transplant. Immunol. 3, 55-61 (1995); Yang
et al., Transplantation 60, 71-76 (1995); Anderson et al., APMIS 102, 23-27
( 1994).
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A related use for compounds of this invention which bind to VLA-4
is in modulating the immune response involved in "graft versus host" disease
(GVHD). See e.g., Schlegel et al., J. Immunol. 155, 3856-3865 (1995).
GVHD is a potentially fatal disease that occurs when immunologically
competent cells are transferred to an allogenetic recipient. In this
situation,
the donor's immunocompetent cells may attack tissues in the recipient.
Tissues of the skin, gut epithelia and liver are frequent targets and may be
destroyed during the course of GVHD. The disease presents an especially
severe problem when immune tissue is being transplanted, such as in bone
marrow transplantation; but less severe GVHD has also been reported in
other cases as well, including heart and liver transplants. The therapeutic
agents of the present invention are used, inter alia, to block activation of
the
donor T-cells thereby interfering with their ability to lyse target cells in
the
host.
A further use of the compounds of this invention is inhibiting tumor
metastasis. Several tumor cells have been reported to express VLA-4 and
compounds which bind VLA-4 block adhesion of such cells to endothelial
cells. Steinback et al., Urol. Res. 23, 175-83 (1995); Orosz et al., Int. J.
Cancer 60, 867-71 (1995); Freedman et al., Leuk. Lymphoma 13, 47-52
(1994); Okahara et al., Cancer Res. 54, 3233-6 ( 1994).
A further use of the compounds of this invention is in treating
multiple sclerosis. Multiple sclerosis is a progressive neurological
autoimmune disease that affects an estimated 250,000 to 350,000 people in
the United States. Multiple sclerosis is thought to be the result of a
specific
autoimmune reaction in which certain leukocytes attack and initiate the
destruction of myelin, the insulating sheath covering nerve fibers. In an
animal model for multiple sclerosis, marine monoclonal antibodies directed
against VLA-4 have been shown to block the adhesion of leukocytes to the
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endothelium, and thus prevent inflammation of the central nervous system
and subsequent paralysis in the animals'6.
Pharmaceutical compositions of the invention are suitable for use in a
S variety of drug delivery systems. Suitable formulations for use in the
present
invention are found in Remington's Pharmaceutical Sciences, Mace
Publishing Company, Philadelphia, PA, 17th ed. ( 1985).
In order to enhance serum half life, the compounds may be
encapsulated, introduced into the lumen of liposomes, prepared as a colloid,
or other conventional techniques may be employed which provide an
extended serum half life of the compounds. A variety of methods are
available for preparing liposomes, as described in, e.g., Szoka, et al., U.S.
Patent Nos. 4,235,871, 4,501,728 and 4,837,028 each of which is
incorporated herein by reference.
The amount administered to the patient will vary depending upon
what is being administered, the purpose of the administration, such as
prophylaxis or therapy, the state of the patient, the manner of
administration,
and the like. In therapeutic applications, compositions are administered to a
patient already suffering from a disease in an amount sufficient to cure or at
least partially arrest the symptoms of the disease and its complications. An
amount adequate to accomplish this is defined as "therapeutically effective
dose." Amounts effective for this use will depend on the disease condition
being treated as well as by the judgment of the attending clinician depending
upon factors such as the severity of the inflammation, the age, weight and
general condition of the patient, and the like.
The compositions administered to a patient are in the form of
pharmaceutical compositions described above. These compositions may be
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sterilized by conventional sterilization techniques, or may be sterile
filtered.
The resulting aqueous solutions may be packaged for use as is, or
lyophilized, the lyophilized preparation being combined with a sterile
aqueous carrier prior to administration. The pH of the compound
preparations typically will be between 3 and 11, more preferably from S to 9
and most preferably from 7 to 8. It will be understood that use of certain of
the foregoing excipients, carriers, or stabilizers will result in the
formation of
pharmaceutical salts.
The therapeutic dosage of the compounds of the present invention
will vary according to, for example, the particular use for which the
treatment is made, the manner of administration of the compound, the health
and condition of the patient, and the judgment of the prescribing physician.
For example, for intravenous administration, the dose will typically be in the
range of about 20 ~g to about 500 ,ug per kilogram body weight, preferably
about 100 ,ug to about 300 ,ug per kilogram body weight. Suitable dosage
ranges for intranasal administration are generally about 0.1 pg to 1 mg per
kilogram body weight. Effective doses can be extrapolated from
dose-response curves derived from in vitro or animal model test systems.
The following synthetic and biological examples are offered to
illustrate this invention and are not to be construed in any way as limiting
the
scope of this invention. Unless otherwise stated, all temperatures are in
degrees Celsius.
zs
EXAMPLES
In the examples below, the following abbreviations have the
following meanings. If an abbreviation is not defined, it has its generally
accepted meaning.
aq or aq. - aqueous
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AcOH - acetic acid
bd - broad doublet
bm - broad multiplet
bs - broad singlet
Bn - benzyl
Boc - N tert-butoxylcarbonyl
BoczO - di-tert-butyl dicarbonate
BOP - benzotriazol-1-yloxy-
tris(dimethylamino)phosphonium
hexafluorophosphate
Cbz - carbobenzyloxy
CHC13 - chloroform
CH~C12 - dichloromethane
(COCI)2 - oxalyl chloride
d - doublet
dd - doublet of doublets
dt - doublet of triplets
DBU - 1,8-diazabicyclo[5.4.0]undec-7-ene
DCC - 1,3-dicyclohexylcarbodiimide
DMAP - 4-N,N dimethylaminopyridine
DME - ethylene glycol dimethyl
ether
DMF - N,N dimethylformamide
DMSO - dimethylsuifoxide
EDC - 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride
Et3N - triethyiamine
Et,O - diethyl ether
EtOAc - ethyl acetate
EtOH - ethanol
eq or eq. - equivalent
Fmoc - N (9-fluorenylmethoxycarbonyl)
FmocONSu - N (9-fluorenylmethoxycarbonyl)-
succinimide
g - grams
h - hour
H,O - water
HBr - hydrobromic acid
HCl - hydrochloric acid
HOBT - 1-hydroxybenzotriazole hydrate
hr - hour
K,C03 - potassium carbonate
L - liter
m - multiplet
MeOH - methanol
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mg - milligram
MgS04 - magnesium sulfate
mL - milliliter
mm - millimeter
mM - millimolar
mmol - millimol
mp - melting point
N - normal
NaCI - sodium chloride
Na2C03 - sodium carbonate
NaHC03 - sodium bicarbonate
NaOEt - sodium ethoxide
NaOH - sodium hydroxide
NH4C1 - ammonium chloride
NMM - N methylmorpholine
Phe - L-phenylalanine
Pro - L-proline
psi - pounds per square inch
PtOz - platinum oxide
q - quartet
quint. - quintet
rt - room temperature
s - ringlet
sat - saturated
t - triplet
t-BuOH - tert-butanol
TFA - trifluoroacetic acid
THF - tetrahydrofuran
TLC or tlc - thin layer chromatography
Ts - tosyl
TsCI - tosyl chloride
TsOH - tosylate
~cL - microliter
In the examples below, all temperatures are in degrees Celcius
(unless otherwise indicated). The following Methods were used to prepare
the compounds set forth below as indicated.
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Method 1
N Tosylation Procedure
N Tosylation of the appropriate amino acid was conducted via the
method of Cupps, Boutin and Rapoport J. Org. Chem. 1985, 50, 3972.
S
Method 2
Meth,~l Ester preparation Procedure
Amino acid methyl esters were prepared using the method of Brenner
and Huber Helv. Chim. Acta 1953, 36, 1109.
Method 3
BOP Coupling Procedure
The desired dipeptide esters were prepared by the reaction of a
suitable N-protected amino acid (1 equivalent) with the appropriate amino
acid ester or amino acid ester hydrochloride (1 equivalent), benzotriazol-1-
yloxy-tris(dimethylamino)phosphonium hexafluorophosphate [BOP] (2.0
equivalent), triethylamine (1.1 equivalent), and DMF. The reaction mixture
was stirred at room temperature overnight. The crude product was purified
flash chromatography to afford the dipeptide ester.
Method 4
H,~drogenatior~ Procedure I
Hydrogenation was performed using 10% palladium on carbon (10%
by weight) in methanol at 30 psi overnight. The reaction mixtures were
filtered through a pad of Celite and the filtrate concentrated to yield the
desired amino compounds.
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Method S
j~vdrolvsis Procedure I
To a chilled (0°C) THF/Hz0 solution (2:1, 5 - 10 mL) of the
appropriate ester was added LiOH (or NaOH) (0.95 equivalents). The
temperature was maintained at 0°C and the reaction was complete in 1 to
3
hours. The reaction mixture was extracted with ethyl acetate and the
aqueous phase was lyophilized resulting in the desired carboxylate salt.
Method 6
ester H, dro ,~i~ Proce ure II
To a chilled (0°C) THF/H20 solution (2:1, 5 - 10 mL) of the
appropriate ester was added LiOH (1.1 equivalents). The temperature was
maintained at 0°C and the reaction was complete in 1 to 3 hours. The
reaction mixture was concentrated and the residue was taken up into H20 and
the pH adjusted to 2 to 3 with aqueous HCI. The product was extracted with
ethyl acetate and the combined organic phase was washed with brine, dried
over MgS04, filtered and concentrated to yield the desired acid.
Method 7
Fster Hydrolysis Procedure III
The appropriate ester was dissolved in dioxane/H,O { 1:1 ) and 0.9
equivalents of 0.5 N NaOH was added. The reaction was stirred for 3 to 16
hours and then concentrated. The resulting residue was dissolved in HZO and
extracted with ethyl acetate. The aqueous phase was lyophilized to yield the
desired carboxylate sodium salt.
Method 8
ulfonylation Procedure I
To the appropriately protected amino acid analog (11.2 mmol),
dissolved in methylene chloride (25m1) and cooled to -78°C was added
the
*rB
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desired sulfonyl chloride (12 rnmol) followed by dropwise addition of
pyridine (2 mL). The solution was allowed to warm to room temperature
and was stirred for 48 hr. The reaction solution was transferred to a 250 mL
separatory funnel with methylene chloride (100 mL) and extracted with 1N
HCI (50 mL x 3), brine (50 mL), and water (100 mL). The organic phase
was dried (MgS04) and the solvent concentrated to yield the desired product.
Method 9
Reductive Amination Procedure
Reductive amination of Tos-Pro-p-NH,-Phe with the appropriate
aldehyde was conducted using acetic acid, sodium triacetoxyborohydride,
methylene chloride and the combined mixture was stirred at room
temperature overnight. The crude product was purified by flash
chromatography.
Method 10
BOC Removal Procedure
Anhydrous hydrochloride (HCl) gas was bubbled through a
methanolic solution of the appropriate Boc-amino acid ester at 0°C for
15
minutes and the reaction mixture was stirred for three hours. The solution
was concentrated to a syrup and dissolved in Et~O and reconcentrated. This
procedure was repeated and the resulting solid was placed under high
vacuum overnight.
Method 11
Tert-Butyl Ester ydrolvsis Pro~gdure I
The tent-butyl ester was dissolved in CH,CI, and treated with TFA.
The reaction was complete in 1-3 hr at which time the reaction mixture was
concentrated and the residue dissolved in H20 and lyophilized to yield the
desired acid.
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Method 12
EDC ~ouDing Procedure I
To a CHZCIZ solution (5-20 mL) of N-(toluene-4-sulfonyl)-L-proline
(1 equivalent), the appropriate amino acid ester hydrochloride (1 equivalent),
N-methylmorpholine (1.1-2.2 equivalents) and 1-hydroxybenzotriazole (2
equivalents) were mixed, placed in an ice bath and 1-(3-
dimethylaminopropyl)-3-ethyl carbodiimide ( 1.1 equivalents) was added.
The reaction was allowed to rise to room temperature and stirred overnight.
The reaction mixture was poured into H,O and the organic phase was washed
with sat. NaHC03, brine, dried (MgS04 or Na~SO~), filtered and
concentrated. The crude product was purified by column chromatography.
Method 13
EDC Coupli~~ Procedure II
To a DMF solution (5-20 mL) of the appropriate N-protected amino
acid (1 equivalent), the appropriated amino acid ester hydrochloride (1
equivalent), Et3N (1.1 equivalents) and 1-hydroxybenzotriazole (2
equivalents) were mixed, placed in an ice bath and 1-(3-
dimethylaminopropyl)-3-ethyl carbodiimide ( 1.1 equivalents) was added.
The reaction was allowed to rise to room temperature and stirred overnight.
The reaction mixture was partitioned between EtOAc and H,O and the
organic phase washed with 0.2 N citric acid, H,O, sat. NaHC03, and brine,
then dried (MgS04 or NaZS04), filtered and concentrated. The crude product
was purified by column chromatography or preparative TLC.
Method 14
Sulfonvlation Procedure I~,
The appropriate sulfonyl chloride was dissolved in CHZCI, and placed
in an ice bath. L-Pro-L-Phe-OMe ~ HCl ( 1 equivalent) and Et3N
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(1.1 equivalent) was added and the reaction allowed to warm to room
temperature and stirred overnight under an atmosphere of nitrogen. The
reaction mixture was concentrated and the residue partitioned between
EtOAc and H20 and the organic phase washed with sat. NaHC03, brine,
dried (MgS04 or Na~S04), filtered and concentrated. The crude product was
purified by column chromatography or preparative TLC.
Method 15
Sulfonvlation Procedure III
To a solution of L-Pro-L-4-(3-dimethylaminopropyloxy)-Phe-OMe
[prepared using the procedure described in Method 10] ( 1 equivalent) in
CHZCIz was added Et3N (5 equivalents) followed by the appropriate sulfonyl
chloride ( 1.1 equivalent). The reaction was allowed to warm to room
temperature and stirred overnite under an atmosphere of nitrogen. The
mixture was concentrated, dissolved in EtOAc, washed with sat. NaHC03
and 0.2 N citric acid. The aqueous phase was made basic with solid
NaHC03 and the product extracted with EtOAc. The organic phase was
washed with brine, dried (MgS04 or Na~S04), filtered and concentrated. The
crude methyl ester was purified by preparative TLC. The corresponding acid
was prepared using the procedure described in Method 7.
Method 16
~3rdrogenation Procedure II
To a methanol (10 -15 mL) solution of the azlactone was added
NaOAc (1 equivalent) and 10% Pd/C. This mixture was placed on the
hydrogenator at 40 psi H,. After 8 - 16 hours, the reaction mixture was
filtered through a pad of Celite and the filtrate concentrated to yield the
dehydrodipeptide methyl ester. The ester was dissolved in dioxane/H~O (5-
10 mL), to which was added 0.5 N NaOH ( 1.05 equivalents). After stirring
for 1- 3 hours, the reaction mixture was concentrated and the residue was
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redissolved in H20 and washed with EtOAc. The aqueous phase was made
acidic with 0.2 N HCI and the product was extracted with EtOAc. The
combined organic phase was washed with brine (1 x 5 mL), dried (MgS04 or
Na2S04), filtered and concentrated to yield the acid as approximately a 1:1
mixture of diastereomers.
Method 17
Tert-Bu~,vl Ester Hydrolysis Procedure II
The tent-butyl ester was dissolved in CH,CIz (5 mL) and treated with
TFA (5 mL). The reaction was complete in 1-3 hours at which time the
reaction mixture was concentrated and the residue dissolved in HZO and
concentrated. The residue was redissolved in Hz0 and lyophilized to yield
the desired product.
Example 1
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-(a-methylbenzyloxy)-L-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-tyrosine methyl ester (see
Example 2 (3)) (785 mg, I.89 mmol) was dissolved in DMF {20 mL) at room
temperature. To this was added K,C03 (1.1 eq, 281 mg) and I-bromoethyl
benzene (1.1 eq, 284 ~L). The reaction was stirred for I2 hours at room
temperature. Ethyl acetate (100 mL) was added, and the organic layer
washed several times with brine (5 x 50 mL). The organic layer was dried
over MgS04. Upon filtration and evaporation of the solvents under reduced
pressure, an oil was isolated. The crude material was purified by elution on
silica gel (EtOAc/hexanes (1:4)). The desired material was isolated in 32%
yield (330 mg, 0.6 mmol). The methyl ester (330 mg. 0.6 mmol) was then
converted to the corresponding acid upon treatment with NaOH (1.1 eq, 27
mg), in MeOH:H,O (1:1) (15 mL), for 4 hours at room temperature. EtOAc
was added as well as water. The aqueous layer was collected and acidified
with IN HC1 to pH 2.5, and reextracted with EtOAc. The organic layer was
4
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dried over MgS04. Upon filtration and evaporation of the solvents under
reduced pressure, a foam was isolated in quantitative yields.
NMR data was as follows:
'H NMR (300 MHz, CDC13): 8 = 7.71 (bd, 2H), 7.34 (m, 7H), 7.20
(m, 1H), 7.01 (m, 2H), 6.80 (d, 2H, J= 8.37 Hz), 5.27 (m, 1H), 4.75 (m, 1H),
4.04 (m, 1H), 3.23-2.93 (m, 4H), 2.42 (s, 3H), 1.85 (m, 1H), 1.60 (d, 3H, J=
6.09 Hz), 1.36-1.26 (m, 3H).
'3C NMR (75 MHz, CDC13): 8 = 174.74, 172.22, 157.53, 145.00,
143.77, 133.42, 130.76, 130.58, 129.14, 128.60, 128.48, 127.94, 126.15,
116.57, 76.39, 62.73, 53.90, 50.09, 37.09, 25.07, 24.52, 22.17.
Mass Spectroscopy: (FAB) 537 (M+H).
Example 2
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-tyrosine
N toluene-4-sulfonyl)-L-proline (1.56 g, 6.93 mmol) was dissolved in
50 mL of DMF, with L-tyrosine methyl ester ( 1.1 eq, 1.76g), Et3N (2.2 eq,
2.0 mL), and BOP reagent {l .l eq, 3.36 g). The ester was isolated in 75%
yield (2.14 g, 5.16 mmol). The ester ( 120 mg, 0.27 mmol) was taken up in a
1:1 mixture of MeOH:H20 (5 mL) with 1.1 eq. of NaOH. The title acid was
isolated in quantitative yield as an oil.
NMR data was as follows:
'H NMR (300 MHz, CD30D): 8 = 7.46 (d, 2H, J= 8.25 Hz), 7.12 (d,
2H, J= 7.83 Hz), 6.77 (d, 2H, J= 8.35 Hz), 6.49 (d, 2H, J= 8.39 Hz), 4.46
(m, 1 H), 3 . 80 (m, 1 H), 3 .11-2.70 (m, 4H), 2.19 (s, 3 H), 1.66 (m, 1 H),
1.31
(m, 3H).
'3C NMR (75 MHz, CD30D): 8 = 177.36, 176.12, 160.30, 149.12,
137.29, 134.79, 134.50, 132.22, 131.50, 119.63, 66.67, 57.90, 54.02, 41.06,
34.57, 28.54, 25.79.
Mass Spectroscopy: (FAB) 417 (M+H).
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Example 3
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-carboxyphenylalanine
Diethyl 2-acetamidomalonate was treated with 4-cyanobenzyl
bromide and NaOEt in EtOH, to give after dilution with HzO, and isolation
of the resulting precipitate, diethyl 2-acetamido-2-(4-cyanobenzyl}malonate.
This product was heated in HCI, and the mixture evaporated to give 4-
carboxy-D,L-phenylalanine hydrochloride (D,L-Phe(4-CO,H)-OH~HCI].
This product was treated with MeOH and HCI gas to give after evaporation,
D,L-Phe(4-COzMe)-OMe~HCI. This product was treated with N (toluene-4-
sulfonyl)-L-Pro-OH, BOP, and NMM in DMF, to give after aqueous workup
and flash chromatography, N (toluene-4-sulfonyl)-L-Pro-D,L-Phe(4-
COZMe)-OMe. This product was treated with NaOH in dioxane and water,
to give after acidification, extraction, drying with MgSO~, filtration and
evaporation the title compound as a clear oil.
NMR data was as follows:
'H NMR (CD30D, 300 MHz): b = 7.86 (d, J= 8.2, 1H), 7.83 (d, J=
8.2, 1 H), 7.74 (d, J = 8.3, 1 H), 7.73 (d, J = 8.3, 1 H), 7.41 {d, J = 8.2,
2H),
7.31 (d, J= 8.2, 1H), 7.21 (d, J= 8.1, 1H), 4.53 (dd, J= 7.1, J= 4.7, O.SH),
4.46 (dd, J= 5.9, O.SH}, 4.07-3.99 (m, IH), 3.58-3.45 (m, 1H), 3.28-3.02 (m,
3H), 2.43 (s, 3H), 1.83-1.43 (m, 4H).
'3C NMR (CD30D w/ CD30Na, 75 MHz): 8 = 179.12, 179.08,
177.5, 177.4, 173.1, 173.0, 145.83, 145.80, 139.6, 139.4, 137.8, 137.7,
134.77, 134.75, 131.0, 130.9, 130.5, 130.4, 129.44, 129.39, 129.1, 129.0,
63.61, 63.57, 57.0, 56.9, 50.74, 50.70, 38.7, 38.6, 31.71, 31.65, 25.2, 25.1,
21.50. 21.49.
Mass Spectroscopy: (+FAB, 3-nitrobenzyl alcohol) 461 (M+H).
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Example 4
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-3-(carboxy)phenylalanine
Substitution of 3-cyanobenzyl bromide for 4-cyanobenzyI bromide,
and following the methods for preparation of Example 3, gave the title
compound as a clear oil.
NMR data was as follows:
'H NMR (CD30D, 300 MHz): 8 = 7.82 (s, O.SH), 7.77 (s, O.SH), 7.73
(d, J= 8.2, 1H), 7.72 (d, J= 8.2, 1H), 7.40 (d, J= 8.2, 2H), 7.37-7.19 (m,
3H), 4.53 (dd, J= 7.5, J=4.9, O.SH), 4.46 (t, J= 5.9, O.SH), 4.05 (dd, J= 8.2,
J= 3.5, O.SH), 3.99 (dd, J= 8.8, J= 3.4, O.SH), 3.48-3.42 {m, O.SH), 3.36-
2.99 (m, 3.SH), 2.42 (s, 3H), 1.88-1.38 (m, 4H).
'3C NMR (CD30D w/ CD30Na, 75 MHz): b = 179.18, 179.16,
177.6, 177.5, 173.04, 172.97, 145.82, 145.81, 139.5, 139.4, 137.81, 137.78,
134.77, 134.75, 132.33, 132.31, 131.03, 130.96, 130.7, 130.6, 129.87,
129.85, 129.4, 129.34, 128.96, 128.4, 63.6, 63.5, 57.0, 56.8, 50.77, 50.76,
38.72, 38.68, 31.7, 31.6, 25.4, 25.1, 21.50. 21.48.
Mass Spectroscopy: (+FAB, 3-nitrobenzyl alcohol) 461 (M+H).
Example 5
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-(2-carboxyphenoxy)-L-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-tyrosine methyl ester (2.14 g,
5.16 mmol) was added to a suspension of sodium hydride, 60% in oil (I.l
eq., 228 mg) in xylenes (50 mL) at 0°C. The reaction mixture was
stirred for
5 minutes and cuprous bromide~dimethyl sulfide complex ( 1.4 eq., 1.48 g)
was added. The reaction mixture was stirred at 23 °C for 0.5 hr. To
this was
added sodium 2-iodobenzoate (1.5 eq., 8.06 mmol), and the reaction mixture
was refluxed for 12 hours. EtOAc ( 100 mL) was added, and the organic
layer washed with NH4C1, 10% HCI, and brine, then dried over MgS04. The
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crude material was eluted on column chromatography (silica gel), with
CHCI3:MeOH (9:1), and isolated as an oil. The acid was prepared by
treatment with NaOH ( 1. I eq), in MeOH:HzO ( 1:1 ) for 4 hours at room
temperature. The diacid was isolated as a foam.
NMR data was as follows:
'H NMR (300 MHz, CDCI3): b = 7.71 (m, 2H), 7.29 (m, 4H), 7.19
(m, 4H), 6.72 (m, 1 H), 4.84 (m, 1 H), 4.13 (m, 1 H), 3 .3 9 (m, 1 H), 3 .11
(m,
3H), 2.43 (s, 3H), 1.89 (m, 1H), 1.48 (m, 3H).
'3C NMR {75 MHz, CDCl3): 8 = 172.67, 157.84, 155.89, 155.04,
145.17, 133.61, 133.19, 133.08, 131.69, 131.02, 130.64, 128.42, 127.87,
124.24, 120.04, 119.61, I 16.12, 62.81, 50.31, 37.28, 30.b9, 24.81, 22.15.
Mass Spectroscopy: (FAB) 553 (M+H).
Example 6
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-O-(benzyl)-L-tyrosine
N (Toluene-4-sulfonyl)-L-Pro-OH was treated with (COC1)z and
DMF in CH~Ch to give, after evaporation, N-(Toluene-4-sulfonyl)-L-Pro-CI.
This product was treated with L-Tyr(Bn)-OH and NaOH in THF and HZO, to
give, after acidification, extraction, drying with MgSOa, and evaporation the
title compound as a clear oil.
NMR data was as follows:
'H NMR (DMSO-db, 300 MHz): 8 = 8.04 (d, J = 8.2, 1 H), 7.70 (d,
J= 8.1, 2H), 7.42-7.21 (m, 6H), 7.15 (d, J= 8.5, 2H), 6.90 (d, J= 8.5, 2H),
5.04 (s, 2H), 4.49-4.42 (m, 1H), 4.13-4.09 (m, 1H), 3.33-3.27 (m, 2H), 3.10-
2.89 (m, 3H), 2.38 (s, 3H), 1.60-1.35 (m, 4H).
'3C NMR (DMSO-db, 75 MHz): 8 = 172.63, 170.8, 157.0, 143.6,
137.2, 133.8, 130.3, 129.8, 129.4, 128.9, 128.4, 127.6, 125.3, 114.4, 69.1,
61.3, 53.4, 49.0, 35.8, 30.4, 23.8, 21Ø
Mass Spectroscopy: (+FAB, 3-nitrobenzyl alcohol) 523 (M+H).
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Example 7
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-(iodo)-L-phenylalanine
Following the experimental procedure described for the synthesis of
Example 8, affords the title compound as a foam.
NMR data was as follows:
'H NMR (300 MHz, DMSO-db): 8 = 8.30 (d, 1H, J= 8.40 Hz), 7.87
(d, 2H, J= 8.50 Hz), 7.78 (d, 2H, J= 8.50 Hz), 7.55 (d, 2H, J= 8.30 Hz),
7.25 (d, 2H, J= 8.30 Hz), 4.63 (m, 1H), 4.26 (m, 1H), 3.20 (m, 4H), 2.53 (s,
3H), 1.75 (m, 4H).
Mass Spectroscopy: (FAB) 543 (M+H).
Example 8
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(methoxy)-phenylalanine
Following the experimental procedure described for Example 1, N-
(toluene-4-sulfonyl)-L-proline (861 mg, 3.2 mmol) was dissolved in 20 mL
of DMF, with Et3N (2.0 eq, 981 ~L), BOP (1.1 eq, 1.55 g), and O-methyl-L-
tyrosine methyl ester~HCl salt (1.1 eq). The methyl ester was isolated in 25%
yield (370 mg, 0.80 mmol). It was then hydrolyzed in a 1:1 solution of
MeOH: H,O (5 mL), with NaOH (1.1 eq, 35 mg). The acid was isolated as a
foam, in 60% yield (216 mg, 0.48 mmol).
NMR data was as follows:
'H NMR (300 MHz, CDC13): b = 7.72 (d, 2H, J= 7.80 Hz), 7.46 (d,
1 H, J = 6.30 Hz), 7.33 (d, 2H, J = 7.20 Hz), 7.26 (d, 2H, J = 7.80 Hz), 7.14
(d, 2H, J= 8.40 Hz), 5.65 (bs, 2H), 4.82 (m, 1H), 4.10 (m, 1H), 3.75 (s, 3H),
3.35-3.01 (m, 4H), 2.42 (s, 3H), 1.97 (m, 1H), 1.48 (m, 3H).
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'3C NMR (75 MHz, CDC13): b = 174.57, 172.25, 159.18, 145.07,
133.40, 130.97, 130.59, 128.66, 128.44, 114.53, 62.78, 55.84, 54.09, 50.20,
37.09, 30.48, 24.68, 22.15.
Mass Spectroscopy: (FAB) 447 (M+H).
Example 9
Synthesis of
N (Toluene-4-sulfony!)-L-prolyl-4-nitro-L-phenylalanine
N (toluene-4-sulfonyl)-L-proline (955 mg, 3.4 mmol) was dissolved
in dry DMF (50 mL) with L-(4-nitro)-phenylalanine methyl ester (l.l eq,
777 mg), Et3N (2.2 eq, 1.04 mL), and BOP reagent (1.1 eq, 1.65 g). The
desired methyl ester was isolated in 40% yield {500 mg, 1.05 mmol). The
ester was hydrolyzed in a 1:1 solution of MeOH:HzO (10 mL) with NaOH
(1.1 eq, 42 mg). The title compound was isolated as an oil in 51% yield (246
mg, 0.53 mmol).
NMR data was as follows:
'H NMR (300 MHz, CDCl3): 8 = 8.10 (d, 2H, J= 8.79 Hz), 7.81 (d,
1 H, J = 8.25 Hz), 7.68 (d, 2H, J = 8.22 Hz), 7.42 (d, 2H, J = 8.79 Hz), 7.32
(d, 2H, J = 8.25 Hz), 4.93 (m, 1 H), 4.09 (m, 1 H), 3.44 (m, 2H), 3.24 (m, 1
H),
3.04 (m, 1H), 2.38 (s, 3H), 1.83 (m, 1H), 1.38 {m, 3H).
'3C NMR (75 MHz, CDC13): b = 174.25, 173.OU, 147.87, 145.34,
145.06, 133.06, 131.13, 130.73, 128.42, 124.18, 62.62, 53.29, 50.04, 37.48,
30.21, 24.35, 21.76.
Mass Spectroscopy: (FAB) 462 (M+H).
Example 10
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-(O-tert-butyl)-L-tyrosine
Following the experimental procedure described in Example 8, the
desired material was isolated as a solid, mp = dec >80°C.
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NMR data was as follows:
'H NMR (300 MHz, CDCl3): 8 = 7.66 (d, 2H, .I= 8.40 Hz), 7.32 (d,
2H, J= 8.01 Hz), 7.09 (d, 2H, J= 8.37 Hz), 6.91 (d, 2H, J= 7.86 Hz), 4.75
(m, 1 H), 4.00 (m, 1 H), 3.25 (m, 2H), 3.05 (m, 2H), 2.40 (s, 3H), 1.95 (m,
4H), 1.27 (s, 9H).
Mass Spectroscopy: (FAB) 489 (M+H).
Example 11
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(3,5-diiodo)-tyrosine
Following the experimental procedure described in Example 8 but
substituting 3,5-diiodo-L-tyrisine methyl ester-HCl salt for the O-methyl-L-
tyrosine provided for the title compound as a foam.
NMR data was as follows:
'H NMR (300 MHz, CDCl3): b = 7.74 (d, 2H, J= 8.19 Hz), 7.51 (s,
1 H), 7.42 (m, 1 H), 7.36 (d, 2H, J = 8.22 Hz), 7.26 (s, 1 H), 4.75 (m, 1 H),
4.12
(m, 1H), 3.55 (m, 1H), 3.24 (m, 2H), 3.05 (m, 1H), 2.44 (s, 3H), 1.62 (m,
4H).
Mass Spectroscopy: (FAB) 685 (M+H), 713 (M+Na).
Example 12
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(aminobenzoyl)-phenylalanine
Following the experimental procedure described for Example l, the
title compound was isolated as an oil in quantitative yields.
NMR data was as follows:
'H NMR (300 MHz, CD30D): b = 7.70 (d, 2H, J= 8.50 Hz), 7.62 (d,
2H, J= 8.50 Hz), 7.44-7.17 (m, 5H), 7.20 (d, 2H, J= 8.25 Hz), 7.06 {d, 2H, J
= 8.52 Hz), 4.47 (m, 1 H), 3.89 (m, 1 H), 3.18 (m, 1 H), 3 .05-2.83 (m, 3H),
2.20 (s, 3H), 1.60-1.39 (m, 4H).
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'3C NMR (75 MHz, CD30D): b = 177.08, 171.91, 148.83, 141.80,
139.37, 138.10, 137.48, 136.92, 134.12, 132.72, 132.37, 131.69, 125.32,
66.34, 58.09, 53.70, 40.85, 34.78, 28.47, 24.61.
Mass Spectroscopy: (FAB) 537 (M+H).
Preparative Example A
Synthesis of
N (Toluene-4-sulfonyl)-D-prolyl-D-phenylalanine
Boc-D-Pro-OH and D-Phe-OBmHCI were treated with BOP and
NMM in DMF, to give after aqueous workup and flash chromatography,
Boc-D-Pro-D-Phe-OBn. This product was treated with TFA and anisole, and
the mixture was evaporated. The residue was dissolved in EtzO and washed
with saturated aqueuos NaHC03 and saturated aqueous NaCI. The Et~O
layers were dried with MgS04, f ltered, and evaporated to give D-Pro-D-Phe-
OBn. This product was treated with CH3SO,CI and Et3N in CHzCIz, to give
after aqueous workup and flash chromatography, N-(CH3S0~)-D-Pro-D-Phe-
OBn. This product was treated with 10% Pd on C in THF, and the mixture
was shaken under 50 psi Hz. The mixture was filtered through Celite, and
evaporated to give the title compound as a solid, mp = 71-75 °C.
NMR data was as follows:
'H NMR (CDCl3, 300 MHz): 8 = 1.24-1.54 (m, 3H), 1.95 (m, 1H),
2.43 (s, 3H), 3.10 (m, 2H), 3.32 (m, 2H), 4.09 (m, 1 H), 4.82 (m, 1 H), 7.10-
7.40 (m, 7H), 7.69 (d, 2H, J = 8.0 Hz).
'3C NMR (CDC13, 75 MHz): 8 = 22.1, 25.8, 32.2, 38.8, 55.4, 63.8,
128.5, 129.5, 130.0, 131.1, 131.6, 135.5, 138.7, 146.3, 174.58, 174.63.
Mass Spectroscopy: (FAB+) 417 (M+H).
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Example 13
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(3-iodo-4-hydroxy)-phenylalanine
The title compound was prepared according to the procedure
described for the synthesis described in Example 8. The corresponding
material was isolated as a film.
NMR data was as follows:
'H NMR (300 MHz, DMSO-db): 8 = 7.72 (d, 2H, J= 8.50 Hz), 7.42
(s, 1H), 7.43 (d, 2H, J= 8.50 Hz), 7.08 (d, 1H, J= 8.05 Hz), 6.75 (d, 1H, J=
8.05 Hz), 4.81 (m, 1 H), 4.11 (m, 1 H), 3.45 (m, 1 H), 3.20 {m, 2H), 2.98 (m,
1 H}, 2.62 (m, 1 H), 2.44 (s, 3H), 1.96 (m, 2H).
Example 14
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-chloro)phenylalanine
The title compound was prepared from the product of Example 37
using the procedure described in Method 7, mp = >200°C
Example 15
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-leucine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-leucine
methyl ester hydrochloride using the procedure described in Method 3. The
title compound was prepared via hydrolysis of the methyl ester using LiOH
in THF/water.
NMR data was as follows:
'H NMR (CDCl3): b = 9.80 (bs, 1H), 7.72 (d, 2H, J= 8.0 Hz), 7.47
(d, 1H, J= $.0 Hz), 7.32 (d, 2H, J= 8.0 Hz), 4.56 (m, 1H), 4.16 (m, 1H),
3.51 (m, 1H), 3.17 (m, 1H), 2.38 {s, 3H), 2.12 (m, 1H), 1.73-1.54 (6H), 0.91
(d, 3H, J= 6.5 Hz), 0.89 (d, 3H, J= 6.0 Hz).
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'3C NMR (CDC13): 8 = 176.2, 172.5, 145.0, 133.4, 130.6 128.4, 62.7,
51.7, 50.3, 41.4, 30.5, 25.5, 23.4, 22.4, 22.1.
Mass Spectroscopy: FAB m/e 383 (M+H).
Example 16
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-alanine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-alanine
methyl ester hydrochloride using the procedure described in Method 3. The
resulting methyl ester was deesterified via hydrolysis using LiOH in
THF/water to provide the title compound as a solid, mp = 160.5-162.5
°C.
NMR data was as follows:
'H NMR (CDCl3): b = 7.76 (d, 2H, .I = 8.2 Hz), 7.52 (d, 1H, J= 6.7
Hz), 7.37 (d, 2H, J = 8.0 Hz), 4.56 (p, 1 H, J = 7.1 Hz), 4.14 (dd, 1 H, J =
2.6,
8.5 Hz), 3.56 (m, 1H), 3.21 (dt, 1H, J= 6.6, 9.7 Hz), 2.45 (s, 3H), 2.21 (m,
1 H), 1.78 (m, 1 H), 1.62 (m, 2H), 1.52 (d, 3 H, J = 7.1 Hz).
'3C NMR {CDC13): 8 = 176.4, 172.3, 145.1, 133.3. 130.6, 128.5, 62.7,
50.4, 49.1, 30.5, 25.0, 22.2, 18.5.
Mass Spectroscopy: FAB m/e 341 (M+H).
Example 17
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(acetamido)-phenylalanine
The title compound was prepared from the product of Example 39
using the procedure described in Method 6.
NMR data was as follows:
'H NMR (300 MHz, CD30D): 8 = 7.50 (d. 2H, J= 8.25 Hz), 7.28 (d,
2H, J= 8.37 Hz), 7.17 (d, 2H, J= 8.20 Hz), 6.99 (d, 2H, J= 8.37 Hz), 4.50
(m, 1 H), 3.92 (m, 1 H), 3.17 (m, 1 H), 3.04 (m, 2H), 2.81 (m, 1 H), 2.19 (s,
3H), 1.87 (s, 3H), 1.56 (m, 1H), 1.40 (m, 3H).
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'3C NMR (7S MHz, CD30D): 8 = 174.61, 172.14, 146.30, 139.29,
135.47, 134.38, 131.64, 131.48, 129.54, 121.71, 63.82, SS.36, S 1.20, 38.27,
32.26, 25.93, 24.50, 22.19.
Mass Spectroscopy: (FAB) 474 (M+H).
S
Example 18
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-isoleucine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-
isoleucine methyl ester hydrochloride using the procedure described in
Method 3. The title compound was prepared via hydrolysis of the methyl
ester using LiOH in THF/water.
NMR data was as follows:
'H NMR (CDCI3): b = 7.72 (d, 2H, J= 8.3 Hz), 7.46 (d, IH, J= 7.S
1 S Hz), 7.36 (d, 2H, J = 8.1 Hz), 4. S4 (dt, 1 H, J = S .4, 7.6 Hz), 4.16
(dd, 1 H, J =
2.6, 8.7 Hz), 3.54 (m, 1 H), 3.21 (m, 1 H), 2.44 (s, 3H), 2.20 (m, 1 H), 1.97
(m,
1H), 1.33-1.57 (4H), 1.35 (m, 4H), 0.90 (d, 3H, J= 7.0 Hz).
''C NMR (CDC13): 8 = 176.2, 172.2, 145.1, 133.4, 130.6, 128.5, 62.8,
53.3, 50.4, 32.1, 30.4, 28.0, 25.1, 22.8, 22.2, 14.5.
Mass Spectroscopy: FAB m/e 383 (M+H).
Example 19
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-aspartic acid
2S l~=(Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-4-(1,1-
dimethylethyl)-aspartic acid methyl ester hydrochloride using the procedure
described in Method 3. The methyl ester was hydrolyzed using LiOH in
THF/water to provide a solid, mp = IS3-1SS°C. The title compound
was
prepared, via cleavage of the t-butyl ester using trifluoroacetic acid in
CHZCI,, as a solid, mp = 174-176°C.
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NMR data was as follows:
'H NMR (DMSO-db): 8 = 8.27 (d, 1H, J= 8.0 Hz), 7.75 (d, 2H, J=
8.2 Hz), 7.44 (d, 2H, J = 8.2 Hz), 4.54 (m, 1 H), 4.16 (m, 1 H), 3 .3 8 (m, 1
H),
3 .13 (m, I H), 2.74 (dd, 1 H, J = 5.9, 16.7 Hz), 2.63 (dd, 1 H, J = 6.3, 16.7
Hz),
S 2.41 (s, 3H), 1.75 (m, 2H), I.51 (m, 2H).
'3C NMR (DMSO-db): 8 = 172.5, 172.2, 171.3, 144.0, 134.3, 130.2,
127.9, 61.7, 49.3, 48.9, 36.2, 30.9, 24.3, 21.4.
Mass Spectroscopy: FAB m/e 385 (M+H).
Example 20
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-lysine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-E-Cbz-
lysine methyl ester hydrochloride using the procedure described in Method 3.
The methyl ester was hydrolyzed using LiOH in THF/water. The title
compound was prepared using the procedure described in Method 4 as a
solid, mp = >200°C.
Example 2I
Synthesis of
N {Toluene-4-sulfonyl)-L-prolyl-L-glutamic acid
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-5-( I , l -
dimethylethyl)-glutamic acid methyl ester hydrochloride using the procedure
described in Method 3. The methyl ester was hydrolyzed using LiOH in
THF/water to provide a solid, mp = 164-166°C. The title compound
was
prepared, via cleavage of the t-butyl ester using trifluoroacetic acid in
CH,CI,, as a solid, mp = >200°C.
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Example 22
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L
(4-dibenzylamino)-phenylalanine methyl ester
The title compound was prepared using the procedure described in
Method 9. The crude product was purified by flash chromatography (silica,
1:1 EtOAc:hexane) to afford the methyl ester as a white solid, mp = 60-
65 ° C.
Example 23
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(N benzyl)-histidine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-(N
benzyl)-histidine methyl ester dihydrochloride using the procedure described
1 S in Method 3. The title compound was prepared via hydrolysis of the methyl
ester using LiOH in THF/water.
NMR data was as follows:
'H NMR (DMSO-db): b = 8.15 (2, IH, J= 7.8 Hz), 7.72 (m, 3H),
7.42 (d, 2H, J = 8.0 Hz), 7.37-7.21 (6H), 6.96 (s, 1 H), 5.11 (s, 2H), 4.42
(m,
1 H), 4.09 (m, 1 H), 3.29 (m, 1 H), 3.04 (m, 1 H), 2.90 (m, 3H), 2.40 (s, 3H),
1.65-1.39 (4H).
'3C NMR (DMSO-db): 8 = 173.1, 171.0, 144.0, 138.0, 137.7, I37.1,
134.2, 130.2, 129.0, 128.9, 128.0, 127.9, 117.3, 61.8, 52.5, 49.9, 49.3, 30.7,
30.1, 24.1, 21.4.
Mass Spectroscopy: FAB m/e 497 (M+H).
Example 24
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-dibenzylamino)-phenylalanine
The methyl ester was prepared using the procedure described in
Example 22. The crude product was purified by flash chromatography
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(silica, 1:1 EtOAc:hexane) to afford the methyl ester as a white solid, mp =
60-65 °C. The title compound was then prepared using the procedure de-
scribed in Method 5.
NMR data was as follows:
'H NMR {DMSO-db, 400 MHz): b = 7.72 (d, 2H, J= 8.34Hz); 7.55
(d, 1 H, J = 5.71 Hz); 7.4 (d, 2H, J = 7.9Hz); 7.24 (m, 4H); 7.18 (m, 6H);
6.76
(d, 2H, J= 8.56Hz); 6.44 (d, 2H, J= 8.56Hz); 4.58 (d, 4H, J= 3.07Hz); 3.89
(dd, 1H, J= 2.74, 8.89Hz); 3.76 (m, 1H); 2.92 (m, 4H); 2.39 (s, 3H); 1.61
(m, 1 H); 1.05-I .34 (m, 3H).
IR (KBr, cm-') 3400, 1655, 1620, 1520, 1405, 1300, 1160.
Mass Spectroscopy: ((+) FAB, m/e {%)) 634 (20 [M+ Li]+); 612 (100
[M+H]+).
Example 25
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-methionine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to methionine
methyl ester using the procedure described in Method 3. The title compound
was prepared via hydrolysis of the methyl ester using LiOH in THF/water.
NMR data was as follows:
'H NMR (CDC13): 8 = 9.05 (bs, 1H), 7.76 (d. 2H, J= 8.2 Hz), 7.69
(d, 1H, J= 7.7 Hz), 7.37 (d, 2H, J= 7.9 Hz), 4.68 (m. 1 H), 4.16 (m, 1H),
3.57 (m, 1 H), 3.20 (m, 1 H), 2.59 (t, 2H, J = 7.4 Hz), 2.45 (s, 1 H), 2.21 (m
3H), 2.19 (s, 3H), 1.79 (m, 1H), 1.63 (m, 2H).
'3C NMR (CDC13): 8 = 174.5, 172.0, 144.5, I 32.6, 130.0, 127.9, 62.2,
51.9, 49.8, 30.8, 30.0, 29.9, 24.5, 21.6, 15.4.
Mass Spectroscopy: FAB m/e 401 (M+H).
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Example 26
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyi-L-serine
The title compound was prepared via hydrogenolysis of the
S corresponding benzyl ether using 10% Pd/C in EtOH/HOAc (20:1 ). The
reaction mixture was filtered through celite and the filtrate was evaporated
in
vacuo to give a residue which was lyophilized from water to give the title
compound as a white solid.
NMR data was as follows:
'H NMR (CDCl3): 8 = 7.97 (d, 1H, J= 7.1 Hz), 7.74 (d, 2H, J= 8.0
Hz), 7.33 (d, 2H, J= 8.4 Hz), 4.62 (m 1H), 4.21-3.94 (2H), 3.60 (m, 1H),
3.17 (m, 1H), 2.41 (s, 3H), 2.07 (m, 1H), 1.88-1.55 (3H).
'3C NMR (CDCl3): 8 = 172.8, 144.4, 132.8, 130.0, 127.8, 62.3, 55.0,
49.8, 30.8, 30.2, 24.4, 21.5.
I S Mass Spectroscopy: FAB m/e 357 (M+H).
Preparative Example B
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-D-aspartic acid
N Benzyloxycarbonyl-D-4-(1,1-dimethylethyl)-aspartic acid was con-
vented to the methyl ester using the procedure described in Method 2. D-4-
(I,1-dimethylethyl)-aspartic acid methyl ester was prepared from the product
of the previous step utilizing the procedure described in Method 4. N
(Toluene-4-sulfonyl)-L-proline hydrate was coupled to the resulting D-4-
(I,1-dimethylethyl)-aspartic acid methyl ester utilizing the procedure
described in Method 3. The methyl ester was hydrolyzed using the
procedure described in Method 6. The product was isolated as a white solid,
mp = 55 °C. The title compound was then prepared using the procedure
described in Method 11. The product was isolated as a white solid, mp =
131-132°C.
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NMR data was as follows;
'H NMR (DMSO-db, 300 MHz ): 8 = 8.19 (d, 1 H, J = 8.0 Hz); 7.73
(d, 2H, J= 7.0 Hz); 7.43 (d, 2H, J= 8.0 Hz); 4.55, (m 1 H); 4.12, (m, 1H);
3.40 {m, 1H); 3.13 (m, 1H); 2.60 (m, 2H); 2.41 (s, 3H); 1.76 (m, 2H); 1.55
(m, 2H).
'3C NMR (DMSO-db, 75 MHz): b = 172.5, 172.2, 171.3, 144.0,
134.2, 130.2, 127.9, 61.8, 49.4, 48.8, 36.2, 30.9, 24.2, 21.4.
Mass Spectroscopy: (PI-FAB) 385, (M+H)+.
Preparative Example C
Synthesis of
N (Toluene-4-sulfonyl)-L-proiyl-D-glutamic acid
N Benzyloxycarbonyl-D-5-(1,1-dimethylethyl)-glutamic acid was
converted to the methyl ester using the procedure described in Method 2. D-
5-(1,1-dimethylethyl)-glutamic acid methyl ester was prepared from the
product of the previous step utilizing the procedure described in Method 4.
N-(Toluene-4-sulfonyl)-L-proline hydrate was coupled to the resulting D-5-
{ 1,1-dimethylethyl)-glutamic acid methyl ester utilizing the procedure
described in Method 3. The methyl ester was hydrolyzed using the
procedure described in Method 6. The product was isolated as a white solid,
mp = 50°C.
The title compound was then prepared using the procedure described
in Method 11. The product was isolated as a white solid, mp = 60°C.
NMR data was as follows:
'H NMR (DMSO-db, 300 MHz): b = 8.12 (d, 1 H, J = 4.0 Hz); 7.73
(m, 2H); 7.43, (m, 2H); 4.25 (m, 1 H); 4.05 (m, 1 H); 3.43 {m, 1 H); 3.15 (m,
1H); 2.40 (s, 3H), 2.45 (m, 2H); 2.02 (m, 2 H); 1.90 - 1.40 (bm, 4 H).
'3C NMR (DMSO-db, 75 MHz): 8 = 174.3, 173.3, 171.6, 144.0,
134.1, 130.3, 127.8, 61.9, 51.4, 49.5, 31.2, 30.3, 26.3, 24.3, 21.4.
Mass Spectroscopy: (PI-FAB) 399, (M+H)T.
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Example 27
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
thiaprolyl-L-(N benzyl)-histidine
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-thiazolidine-4-carboxylic
acid was prepared from L-(5,5-dimethyl)-thiazolidine-4-carboxylic acid
using the procedure described in Method 1. The title compound was then
prepared using standard coupling procedures to provide a solid, mp =
>200°C (dec.).
I O NMR data was as follows:
'H NMR (D,O, 300 MHz): 8 = 0.95 (s, 3H), 1.08 (s, 3H), 2.41 (s,
3H), 2.85-3.12 (m, 2H), 3.84 (s, 1H), 4.32 (m, 1H), 4.40 (s, 2H), 5.09 (s,
2H), 7.01 (s, 1H), 7.24-7.43 (m, 7H), 7.60-7.71 (m, 3H).
'3C NMR (DZO, 75 MHz): 8 = 27.9, 30.2, 30.3, 35.6, 37.0, 57.7, 61.3,
61.7, 79.8, 124.9, 134.6, 135.06, 135.10, 135.8, 137.1, 138.9, 143.2, 143.4,
143.5, 152.6, 176.7, 184.1.
Mass Spectroscopy: (FAB+) 565 (M+H).
Example 28
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(1-methyl)histidine
The ester of the title compound was synthesized as described in
Method 12. The ester was dissolved in dioxane/H~0 (8 mL) to which was
added solid NaOH (1 eq.). After stirring overnight, the reaction mixture was
concentrated. The residue was redissolved in H20 and loaded onto an ion
exchange column (Dowex SOW-X8-100, H+ form). After washing the
column with H,O, the compound was eluted with 5% aqueous pyridine.
Fractions containing the compound were pooled and concentrated and the
residue was dissolved in H20 and lyophilized. The product was isolated as a
white solid, mp = 135-137°C.
NMR data was as follows:
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'H NMR(DMSO-db, 300 MHz): b = 8.16 (d, 2H, J= 8.0 Hz); 7.72 (d,
2H, J = 8.0 Hz); 7.54 (s, 1 H); 7.42 (d, 2H, J = 8.0 Hz); 6.91 (s, 1 H); 5.10
(bs, 1 H); 4.41 (m, 1H); 4.11 (M, 1H); 3.57 (s, 3H), 3.34 (m, IH); 3.10 (m,
1H); 2.92 (m, 2 H); 2.39 (s, 3 H); 1.80-1.45 (bm, 4H).
'3C NMR(DMSO-db, 75 MHz): b = 173.0, 171.0, 143.9, 137.5, 136.9,
134.3, 130.2, 127.8, 118.6, 61.8, 52.5, 49.3, 33.3, 30.8, 29.9, 24.2, 21.4.
Mass Spectroscopy: (PI-FAB) 421, (M+H)+.
Example 29
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-D-(N benzyl)histidine
A CHZCI, solution of Boc-D-(N benzyl)histidine was chilled to
-15°C (dry ice/CH3CN bath) to which was added diethylisopropylamine
(1.5
eq.), methanol (3.0 eq.) and BOP (1.1 eq) to form the methyl ester. The
reaction was allowed to warm to room temperature and stirred overnight. The
reaction mixture was then poured into 0.1 M HCI and the organic phase
washed with H20, saturated NaHC03, brine, dried (MgSO~), filtered and
concentrated. The crude methyl ester was purified by column
chromatography. The Boc group was removed with TFA/CH,CIz. The crude
reaction mixture was taken-up in CH,CI, and washed with 5% Na,C03. The
organic phase was washed with brine and dried (MgSO~), filtered and
concentrated to give the free amine. The coupling was performed using the
procedure described in Method 12 and the ester hydrolyzed using the
procedure described in Method 7. The product was isolated as a white solid,
mp = >200 ° C.
NMR data was as follows:
'H NMR (DMSO-db, 300 MHz): 8 = 7.75 - 7.61 (m, 3 H); 7.51 (s,
1H); 7.45-7.15 (bm, 7 H); 6.89 (s, 1H); 5.05 (m, 2H}; 4.05 (m, 2H); 3.30 (m,
1 H); 3.10 (m, 1 H); 2.95 (m, 1 H); 2.75 (m, 1 H); 2.38 (s, 3H); 1.67 (m, 2H);
1.35 (m, 2H).
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'3C NMR (DMSO-db, 75 MHz): 8 = 17403, 169.6, 143.9, 139.3,
138.3, 136.5, 134.1, 130.3, 128.9, 127.8, 127.7, 117.1, 62.4, 54.3, 49.8,
49.4,
31.8, 30.6, 24.2, 21.4.
Mass Spectroscopy: (PI-FAB) 519, (M+H)+.
Example 30
Synthesis of
N (Toluene-4-sulfonyl)-L-glutamyl-L-tyrosine
Cbz-L-5-(1,1-dimethylethyl)-glutamic acid was coupled with L-
tyrosine t-butyl ester using the procedure described in Method 3. The Cbz
group was removed using the procedure described in Method 4. The
resulting ester was tosylated using the procedure described in Method 1. The
title compound was prepared using the procedure outlined in Method 11
which provided a solid, mp = 173-175 °C.
NMR data was as follows:
'H NMR (DMSO-db, 300 MHz): b = 1.70-1.90 (m, 2H), 2.24-2.32
(m, 2H), 2.40 (s, 3H), 2.80-3.10 (m, 2H), 3.51 (m, 1 H), 4.0-4.5 (C-H buried
under large Hz0 peak), 6.87 (d, 2H, J= 8.4 Hz), 7.18 (d, 2H, J= 8.2 Hz),
7.44 (d, 2H, J= 8.2 Hz), 7.70 (d, 2H, J= 8.2 Hz), 8.34 (bs, 1H).
"C NMR (DMSO-db, 75 MHz): 8 = 21.5, 28.5, 31.1, 36.7, 52.9, 54.8,
121.9, 128.5, 130.6, 131.1, 131.9, 138.0, 146.1, 147.9, 170.7, 173.0, 174.7.
Mass Spectroscopy: (FAB+) 465 (M+H).
Example 31
Synthesis of
N {Toluene-4-sulfonyl)-L-prolyl-L-(N 3-methyl)histidine
Boc-L-(3-methyl)histidine was dissolved in MeOH and chilled in an
ice bath. HCl gas was then bubbled through the reaction mixture for i 5
minutes. After stirring overnight, the reaction mixture was concentrated to a
gummy solid. The dihydrochloride was coupled to Tos-Pro-OBn using the
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procedure described in Method 12 and hydrolyzed via Method 7. The
product was isolated as a white solid, mp = >200 °C.
NMR data was as follows:
'H NMR (DMSO-db, 300 MHz): b = 7.74 (m, 3H); 7.45 - 7.35 (bm,
3H); 6.56 (s, 1H); 4.10-3.93 (m, 2H); 3.51 (s, 3 H), 3.22 (m, IH), 3.10 (m,
2H); 2.91 (m, 1H); 2.38 (s, 3 H); 1.70 (m, 1H); 1.45 (m, 3H).
'3C NMR (DMSO-db, 75 MHz): b = 173.3, 170.1, 144.1, 137.5,
133.9, 130.3, 128.8, 128.1, 127.5, 62.4, 53.6, 49.1, 31.2, 30.6, 26.2, 24.1,
21.4.
Mass Spectroscopy: (PI-FAB) 443, (M+H)+.
Example 32
Synthesis of
N {Toluene-4-sulfonyl)-L-prolyl-a-amino-2,3-
dihydro-{1,4-benzodioxin)-6-propanoic acid
The title compound was prepared using the procedure described in
Method 16 and isolated as a white solid.
NMR data was as follows:
'H NMR (CDCI3, 300 MHz): b = 7.73 (m, 2H); 7.32 (m, 2H); 6.87-
6.26 (m, 2 H); 6.64 (m, 1 H); 4.84 (m, I H); 4.21 (m, 4H), 4.10 (m, 1 H); 3.55-
3.40 (m, 1H); 3.30-2.95 (m, 3H); 2.43 (m, 3H); 2.06 (m, 1 H); 1.64 (m, 3H).
'3C NMR (CDCl3, 75 MHz): 8 = 175.0, 174.5, 172.4, 172.3, 145.0,
144.9, 144.2, 143.9, 143.4, 143.2, 133.5, 133.4, 130.6, 129.6, 129.1, 128.5,
123.2, 122.69, 118.9, 118.7, 118.1. 117.9, 64.9, 63.1, 62.8, 53.9, 53.8, 50.4,
50.2, 37.3, 37.2, 30.9, 30.5, 24.8, 24.7, 22.2.
Mass Spectroscopy: (PI-FAB) 475, (M+H)+.
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Example 33
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
a-amino-1,3-benzodioxole-5-propanoic acid
The title compound was prepared using the procedure described in
Method 16 and isolated as a white solid.
NMR data was as follows:
'H NMR (CDCl3, 300 MHz): b = 7.71 (m, 2H); 7.50 - 7.29 (bm, 3H);
7.81-7.60 (m, 3H); 5.90 (m, 2H); 4.80 (m, 1 H); 4.15 (m, 1 H); 3.43 (m, 1 H);
3.30-3.00 (bm, 3H); 2.41 (s, 3H); 2.10 (m, 1 H); 1.70 (m, 1 H); 1.51 (m, 2H).
'3C NMR (CDCl3, 75 MHz): 8 = 174.9, 174.5, 172.5, 172.3, 148.5,
148.2, 147.4, 147.2, 145.0, 133.4, 133.2, 130.6, 130.2, 129.5, 128.5, 128.5,
123.6, 123.1, 110.5, 110.2, 109.2, 108.8, 101.5, 63.1, 62.7, 54.0, 53.7, 50.4,
50.2, 37.8, 37.6, 31.0, 30.4, 24.8, 22.1.
1 S Mass Spectroscopy: (PI-FAB) 461, (M+H)+.
Example 34
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-valine
N (Toluene-4-sulfonyl)-L-proline hydrate was coupled to L-valine
methyl ester hydrochloride using the procedure described in Method 3. The
title compound was prepared via hydrolysis of the methyl ester using LiOH
in THF/water.
NMR data was as follows:
'H NMR (CDCl3): b = 8.20 (br s, 1 H), 7.76 (d, 2H, J = 8. Hz), 7.49
(d, 1H, J = 8.3 Hz), 7.35 (d, 2H, J = 8.0 Hz), 4.50 (dd, 1H, J = 4.8, 8.3 Hz),
4.19 (m, 3H), 3.54 (m, 1H), 3.21 (m, 1H), 2.44 (s, 3H), 2.31-2.18 (2H), 1.70-
1.56 (3H), 1.00 (d, 3H, J = 6.9 Hz), 0.99 (d, 2H, J = 6.9 Hz).
'3C NMR (CDCl3): b = 175.7, 172.3, 145.0, 133.5, 130.6, 128.5, 62.8,
58.1, 50.3, 31.6, 30.3, 25.1, 22.2, 19.7, 18.2.
Mass Spectroscopy: FAB m/e 369 (M+H).
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Example 35
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-iodo)-phenylalanine methyl ester
The procedure used for the preparation of Example 8 was utilized.
The title compound was isolated as a white foam.
NMR data was as follows:
'H NMR (300 MHz, CDC13): b = 7.72 (d, 2H, J = 8.25 Hz), 7.64 (d,
2H, J = 8.25 Hz), 7.41 (d, 1 H, J = 8.20 Hz), 7.36 (d, 2H, J = 8.10 Hz), 7.00
(d, 2H, J = 8.25 Hz), 4.82 (m, 1H), 4.08 (m, 1H), 3.71 (s, 3H), 3.35 (m,2H),
3.10 (m, 2H), 2.44 (s, 3H), 2.01 (m, 1H), 1.55 (m, 3H).
Mass Spectroscopy: (FAB) 557 (M+H).
Example 36
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-
L-4-(aminobenzoyl)phenylalanine methyl ester
Followed the experimental section described for Example 1. N-
(Toluene-4-sulfonyl)-L-prolyl-L-p-amino-phenylalanine methyl ester { 1.75
g, 3.93 mmol) was dissolved in dichloromethane (25 mL) with Et3N ( 1.1 eq,
600 mL) and benzoyl chloride (1.1 eq, 860 mL). The title material was
isolated as a solid, in 90% yield (1.95 g, 3.55 mmol), mp = 191-193 °C.
NMR data was as follows:
'H NMR (300 MHz, CDCl3): 8 = 7.94 (broad s, 1 H), 7.67 (d, 2H, J =
8.31 Hz), 7.71 (d, 2H, J = 8.22 Hz), 7.60 (d, 2H, J = 8.37 Hz), 7.45 (m, 5H),
7.34 (d, 2H, J = 8.34 Hz), 7.16 (d, 2H, J = 8.25 Hz), 4.82 (m, 1 H), 4.40
(broad s, 1 H), 4.05 (m, 1 H), 3.77 (s, 3H), 3.40 (m, l H), 3.23 (m, 1 H),
3.09
(m, 2H), 2.42 (s, 3H), 2.02 (m, 1H0, 1.56 (m, 3H).
'3C NMR (75 MHz, CDC13): b = 171.93, 171. 46, 144.93, 137.98,
135.45, 133.49, 132.78, 132.44, 130.55, 129.36, 128.43, 127.53, 120.80,
62.83, 54.02, 53.11, 50.29, 37.96, 30.31, 24.88, 22.16.
Mass Spectroscopy: (FAB) 550 (M+H).
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Example 37
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-chloro)phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-proline (660 mg, 2.3 mmol) was added to
12 mL of DMF, at room temperature under Nz. To this was added L-(4-
chloro)phenylalanine methyl ester hydrochloride ( 1.1 eq. 493.3 mg), N
methyl morpholine (3.5 eq., 890 mL) and BOP reagent (1.0 eq., 660 mg).
The title compound was isolated as an oil in 31 % yield (310 mg, 0.7 mmol).
NMR data was as follows:
'H NMR (300 MHz, CDCl3): 8 = 7.70 (broad d, 2H), 7.35 (m, 2H),
7.25 (m, 2H), 7.11 (m, 2H), 4.82 (m, 1 H), 4.05 (m, 1 H), 3.78 (s, 3H), 3.35
(m, 1H), 3.25 (m, 1H), 3.05 (m,2H), 2.45 (s,3H), 1.52 (m, 3H).
Mass Spectroscopy: (FAB) 433 (M+H).
Example 3 8
Synthesis of
N {Toluene-4-sulfonyl)-L-prolyl-L-(4-amino)phenylalanine methyl ester
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-nitro)phenylalanine methyl
ester (2.00 g, 4.48 mmol) was dissolved in MeOH ( 10 mL) with a catalytic
amount of 10% Pd on C. The hydrogenation reaction was run at room
temperature for 12 hours at 40 psi. Upon filtration of the reaction mixture
over Celite, the solvent was evaporated under reduced pressure yielding the
title compound as a pink foam in quantitative yields.
NMR data was as follows:
'H NMR (300 MHz, CDC13): b = 7.68 (d, 2H, J = 8.25 Hz), 7.30 (d,
2H, J = 8.07 Hz), 6.88 (d, 2H, J = 8.25 Hz), 6.57 (d, 2H, J = 8.25 Hz), 4.74
(m, 1H), 4.04 (broad m, 3H), 3.70 (s, 3H), 3.36 (m, 1H), 3.11 (m, 2H), 2.92
(m, 1H), 2.39 (s, 3H), 1.97 (m, 1H), 1.48 {m, 3H).
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'3C NMR (75 MHz, CDC13): b = 172.14, 171.31, 145.97, 144.86,
133.61, 130.65, 130.51, 128.43, 126.23, 115.79, 111.35, 62.83, 54.16, 52.96,
50.21, 37.68, 30.34, 24.76, 22.16, 14.7.
Mass Spectroscopy: (FAB) 446 (M+H).
Example 39
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L
(4-acetamido)phenylalanine methyl ester
The title compound was prepared following the experimental proce-
dure described for Example l and was isolated as a foam in quantitative
yields.
NMR data was as follows:
'H NMR (300 MHz, CDCl3): b = 7.70 (d, 2H, J = 8.10 Hz), 7.65 (m,
2H), 7.33 (d, 2H, J = 8.40 Hz), 7.08 (d, 2H, J = 8.40 Hz), 4.79 (m, 1H), 4.05
(m, 1H), 3.75 (s, 3H), 3.37 (m, 1H), 3.11 (m, 3H), 2.42 (s, 3H), 2.22 (s, 3H),
2.02 (m, 1H), 1.48 (m, 3H).
"C NMR (75 MHz, CDC13): S = 177.61, 171.92, 171.46, 169.07,
144.98, 137.69, 133.41, 132.25, 130.56, 130.32, 128.41, 120.40, 62.83,
54.00, 53.10, 50.28, 45.10, 37.89, 30.34, 25.13, 24.84, 22.17.
Mass Spectroscopy: (FAB) 488 (M+H).
Example 40
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiaprolyl-L-
(N benzyl)-histidine methyl ester
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)thiazolidine-4-carboxylic
acid was prepared from L-(5,5-dimethyl)thiazolidine-4-carboxylic acid using
the procedure described in Method 1. The title compound was prepared
using standard coupling procedures to provide a solid, mp = 60-66°C.
NMR data was as follows:
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'H NMR (CDC13, 300 MHz): 8 = 1.17 (s, 3H), 1.30 (s, 3H), 2.43 (s,
3H), 3.09 (m, 2H), 3.67 (s, 3H), 3.92 (s, 1H), 4.45 (d, 1H, J = 10.2 Hz), 4.62
(d, 1 H, J = 10. I Hz), 5.03 (s, 2H), 6.71 (s, 1 H), 7.13 (d, 2H, J = 7.7 Hz),
7.29-7.3 8 (m, SH), 7.46 (s, 1 H), 7.76 (d, 2H, J = 8.1 Hz), 7.93 (d, 1 H, J =
7.4
Hz).
'3C NMR (CDC13, 75 MHz): 8 = 22.3, 24.9, 29.8, 30.3, 51.1, 51.5,
52.9, 53.0, 55.3, 74.0, 117.9, 128.0, 128.6, 128.9, 129.6, 130.5, 133.0,
136.5,
137.6, 137.9, 145.0, 169.1, 171.9.
Mass Spectroscopy: (FAB+) 557 (M+H).
Example 4
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-(3-3'-tolylureido)-L-phenylalanine
The methyl ester was prepared by the reaction of N (toluene-4-
sulfonyl)-L-prolyl-L-4-aminophenylalanine with 3-tolylisocyanate
(triethylamine, toluene, reflux one hour). The resulting solid was removed
by filtration and washed well with EtOAC/Hexane to give the methyl ester as
a white solid. The methyl ester was hydrolyzed using 1 M LiOH in THF.
The title compound was isolated following acid/base work-up as a white
solid, mp = 135-140°C.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): S = 12.79 (br s, 1H); 8.55 {s, 1H);
8.50 (s, 1H); 8.02 (d, 1H, J = 7.9Hz); 7.69 (d, 2H, J = 8.3Hz); 7.39 (d, 2H, J
= 7.9Hz); 7.33 (d, 2H, J = 8.56Hz); 7.26 (s, 1 H); 7.18 (d, 1 H, J = 8.4Hz);
7.12 (m, 3H); 6.75 (d, 1H, J = 7.25Hz); 4.43 (m, IH); 4.11 (dd, IH, J = 3,
8.1 Hz); 3.32 (m, 1 H); 3.1 (m, 1 H); 3.01 (dd, l H, J = 5.05, 13.83Hz); 2.91
(dd,
1H, J = 8.45,13.73Hz);2.38 (s,3H); 1.4-1.63 (m, 4H).
IR (KBr, cm'): 3340, 3290, 3090, 2980, 1730, 1650, 1600, 1550,
1495, 1350, 1290, 1210, 1160, 660.
Mass Spectroscopy: ( (+) FAB, m/e (%)) 565 (60 [M+H]+),
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Example 42
Synthesis of
N (Toluene-4-sulfonyl-L-prolyl-4-[(2,3,3a,7a-tetrahydro
1H-indole-2-carbonyl)-amino]-L-phenylalanine
The N Boc compound was prepared using the procedures described
in Method I3 and Method 6. The title compound was then prepared using
the procedure described in Method 10.
NMR data was as follows:
'H NMR (DMSO-d6): b = 7.75 (d, 2H), 7.65 (m, 2H), 7.45 (d, 4H),
7.10 (m, 4H), 6.80 (d, 2H), 4.35 (m, 1 H), 4.10 (b, 1 H), 4.02 (m, 1 H), 3.10
(m, 4H), 2.40 (s, 3H), 1.70 (b, 1H), 1.41 (m, 2H).
Mass Spectroscopy: (+) FAB (M+H)+ 579
Example 43
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L
(4-pentylamino)phenylalanine methyl ester
The methyl ester was prepared by reductive amination of N-(toluene-
4-sulfonyl)-L-prolyl-L-(4-amino)phenylalanine with valeraldehyde (acetic
acid, sodium triacetoxyborohydride, methylene chloride, which was stirred at
room temperature for one hour). The crude product was purified by flash
chromatography (silica, I :1 EtOAc:hexane) to afford the methyl ester as a
white solid, mp = >160°C.
Example 44
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-O
(2-dibenzylamino-ethyl)-tyrosine methyl ester
The title compound was prepared via O-alkylation of N (toluene-4-
sulfonyl)-L-prolyl-L-tyrosine methyl ester with N {2-
chloroethyl)dibenzylamine hydrochloride in refluxing 2-butanone in the
presence of potassium carbonate and sodium iodide. The obtained mixture
*rB
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was chromatographed on Si02 in 1 % MeOH/CHCl3 (Rf = 0.75 using 10%
MeOH/CHC13).
NMR data was as follows:
'H NMR (DMSO-db, 400 Mhz): 8 = 1.4-1.6 (4H, m); 2.38 (3H, s);
S 2.75 (2H, t, J=7,7Hz); 2.95 (2H, m); 3.1 ( 1 H, dd, J=6,10,6Hz); 3.33 ( 1 H,
dd);
3.6 (3H, s); 3.65 (4H, s); 4.0 (2H, t, J=6,6Hz); 4.1 ( 1 H, t, J=8,8Hz); 4.5 (
1 H,
t, J=8,8Hz); 6.75 (2H, d, J=IOHz); 7.08 (2H, d, J=IOHz); 7.2-7.35 (lOH, m);
7.4 (2H, d, J=IOHz); 7.68 (2H, d, J=IOHz); 8.1 (1H, d, J=IOHz).
MS: +FAB, m/z 670 ([MH]+, 45 %), 154 (100%).
Example 45
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-O-[2-(dibenzylamino)ethyl]-tyrosine
The methyl ester was prepared via O-alkylation of N (toluene-4
sulfonyl)-L-prolyl-L-tyrosine methyl ester with N (2-
chloroethyl)dibenzylamine hydrochloride in refluxing 2-butanone in the
presence of potassium carbonate and sodium iodide. The title compound
was prepared using the procedure described in Method 7, and was isolated as
a solid, mp = 106-110°C.
NMR data was as follows:
'H NMR (DMSO-db, 400 Mhz): 8 = 1.2-1.45 (3H, m); 1.75 (1H, m);
2.38 (3H, s); 2.75 (2H, t, J=6,6Hz); 2.95 (2H, m); 3.1 (1H, dd, J=7,6,7Hz);
3.38 (1H, bs); 3.62 (4H, s); 3.85 (1H, dd, J=6,4,6Hz); 3,95 (1H, t, J=4,4Hz);
3.97 {2H, t, J=8,8Hz); 6.62 (2H, d, J=IOHz); 6.95 (2H, d, J=IOHz); 7.2-7.4
(IOH, m); 7.41 (2H, d, J=IOHz); 7.62 (1H, d, J=4Hz); 7.75 (2H, d, J=lOHz).
MS: +FAB, rn/z 656 ([MH]+, 15 %), 154 (100%).
*rB
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Example 46
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-pentylamino)phenylalanine
The title compound was prepared from the product of Example 43
using the procedure described in Method 6.
NMR data was as follows:
'H NMR (DMSO-d6, 400 MHz): b = 7.73 (d, 2H, J = 8.lHz); 7.57 (d,
1 H, J = 5.7Hz); 7.39 (d, 2H, J = 7.9Hz); 6.76 (d, 2H, J = 8.3Hz); 6.32 (d,
2H,
J = 8.SHz); 5.14 (t, 1 H, J = 5.6Hz); 3.94 (dd, 1 H, J = .2,.007Hz); 3.77 (m,
1H); 3.18 (m, 2H); 2.90 (m, 4H); 2.38 (s, 3H); 1.73 (m, 1H); 1.41 (m, SH);
1.29 (m, 4H); .85 (m, 3H).
IR (KBr, cm'): 3400, 2950, 2910, 2870, 1660, 1620, 1525, 1405,
1350, 1155, 670, 600, 550.
Mass Spectroscopy: ( (+) FAB, m/e (%)) 508 (60 [M+ Li)+).
Example 47
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4
(4-chlorobenzylamino)-phenylalanine methyl ester
The title compound was prepared by reductive amination of N
(toluene-4-sulfonyl)-L-prolyl-L-(4-amino)-phenylalanine methyl ester with
4-chlorobenzaldehyde (acetic acid. sodium triacetoxyborohydride, methylene
chloride, stirred at room temperature for 2.5 hours). The crude product was
purified by flash chromatography (silica, 1:1 EtOAc:hexane) to afford the
title compound as a white solid.
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Example 48
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[3
(4-cyanophenyl)-ureido]-phenylalanine methyl ester
The title compound was prepared by the reaction of N (toluene-4-
sulfonyl)-L-prolyl-L-(4-amino)-phenylalanine methyl ester with 4-
cyanophenylisocyanate (triethylamine, toluene, reflux 3.5 hours). The crude
product was purified by flash chromatography (silica, 1:1 EtOAc:hexane) to
give the methyl ester as a white solid, mp = 204-206°C.
Example 49
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[3-
(4-cyanophenyl)-ureido]-phenylalanine
The title compound was prepared from the product of Example 48
using the procedure described in Method 6 as a solid, mp = 163-165 °C.
NMR data was as follows:
' H NMR (DMSO-db, 400 MHz): b = 12.8 (br s, 1 H); 9.12 {s, 1 H);
8.78 (s, 1 H); 8.04 (d, 1 H, J = 8.1 Hz); 7.69 (m,4H); 7.6 (d, 2H, J = 9Hz);
7.39
(d, 2H, J = 8.lHz); 7.35 (d, 2H, J = 8.SHz); 7.15 (d, 2H, J = 8.SHz); 7.12 {m,
3 H); 4.43 (m, 1 H); 4.11 (dd, 1 H, J = 3.1, 8.2Hz); 3.29 (m, 1 H); 3.06 (m,
2H);
2.92 (dd,lH, J = 8.4, 13.9Hz); 2.38 (s,3H); 1.38-1.63 (m, 4H).
IR (KBr, cm'): 3360, 2220, 1720, 1650, 1590, 1530, 1510, 1410,
1230, 1150,1090, 830, 670, 595, 550.
Mass Spectroscopy: ( (-) FAB, m/z (%)) 574 (40 [M-H]-).
Example 50
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-O-(tert-
butoxycarbonylmethyl)-tyrosine methyl ester
The title compound was prepared by reaction of N (toluene-4-
sulfonyl)-L-prolyl-L-tyrosine methyl ester with t-butyl bromoacetate
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(potasiurn carbonate, DMF, under Argon for 72 hrs). The product was
purified by flash column chromatography (silica, 1:1 hexane:EtOAc) to
afford the methyl ester as a white solid, mp = 55 °C.
Example 51
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-O-
(tent-butoxycarbonylmethyl)-tyrosine
The title compound was prepared from the product of Example 50
using the procedure described in Method 6 and was isolated as a solid, mp =
69-70 ° C.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): ~ = 12.2 (br s, 1 H); 8.00 (d, 1 H, J =
8.lHz); 7.67 (d, 2H, J = 8.2Hz); 7.38 (d, 2H, J = 7.9Hz); 7.11 {d, 2H, J =
8.6Hz); 6.76 (d, 2H, J = 8.SHz); 4.5 (s, 2H); 4.4 (m, 1 H); 4.07 (m, 1 H); 2.8-
3.1 {m, 4H); 2.37 (s, 3H); 1.5 (m, 4H); 1.38 (s, 9H).
IR (KBr, cm'): 3400, 2950, 1750, 1660, 1510, 1340, 1225, 1160,
1075, 660, 575, 525.
Mass Spectroscopy: (ESI, m/e (%)) 545 ( 100, (M-H)).
Example 52
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-[(3S)-3,
4-dihydro-isoquinolin-3-yl-aminocarbonyl)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-amino)phenylalanine methyl
ester (20 mmol) was taken into CH~C12 with HOBT, Boc-L-tetrahydro-
isoquinoline-3-carboxylic acid (20 mmol), and DCC (24.3 mmol). The
mixture was stirred overnight at room temperature, filtered and concentrated
in vacuo. The resulting residue was partitioned between 10% citric acid and
EtOAc. The organic layer was washed with H,O, saturated NaHC03, and
H,O, dried over Na,S04 and concentrated in vacuo to yield an amorphous
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solid. The resulting ester was converted to the acid using the procedure
described in Method 6. The title compound was then prepared using the
procedure described in Method 10 and was isolated as a solid, mp = 145-
I50°C.
NMR data was as follows:
'H NMR (DMSO-db): 8 = 8.12 (d, 1H), 7.70 (d, 2H), 7.55 (d, 2H),
7.40 (d, 2H), 7.25 (d, 6H), 4.45 (m, 1 H), 4.3 8 (d, 1 H), 4.28 (m, 1 H), 4.10
(d,
1 H), 3.10 (m, 2H), 2.40 (s, 3H), 1.60 (m, 4H).
Mass Spectroscopy: (FAB) (M+H)+ 591 (M + Na)+ 613.
Example 53
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-[3
(3-methoxy-phenyl)-ureido]-L-phenylalanine methyl ester
The title compound was prepared by the reaction of N (toluene-4-
sulfonyl)-L-prolyl-L-(4-amino)-phenylalanine methyl ester with 3-methoxy
phenylisocyanate (triethylamine, toluene, reflux one hour). The resulting
solid was removed by filtration and washed with EtOAc/hexane to give the
methyl ester as a white solid, mp = 206-209°C.
Example 54
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-4-[3
(3-methoxy-phenyl)-ureido]-L-phenylalanine
The title compound was prepared from the product of Example 53
using the procedure described in Method 6.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): b = 12.80 (br s, IH); 8.b (d, 2H. J =
13.8); 8.03 (d, 1H, J = 7.9Hz); 7.69 (d, 2H, J = 8.IHz); 7.39 (d, 2H, J =
8.lHz); 7.33 (d, 2H, J = 8.56Hz); 7.14 (m, 1H); 6.9 (dd, 1H, J = 1.53,7.9Hz);
6.52 (dd, 1 H, J = 2.19, 8.12Hz); 4.42 (m, 1 H); 4.11 (dd, 1 H, J = 2.96,
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8.45Hz); 3.7 (s, 3H); 3.35 (m, 1H); 3.09 (m, 1H); 3.01 (dd,lH, J = 4.83,
13.6Hz); 2.91 (dd, 1H, J = 8.34,13.8Hz);2.48 (s,3H); 1.41-1.63 (m, 4H).
IR (KBr, cm'): 3390, 2940, 1655, 1595, 1545, 1500, 1450, 1345,
1320, 1210, 1160, 660, 580, 545.
Mass Spectroscopy: ( (+) FAB, m/e (%)) 603(30[M+Na)+); 581 (10
[M+H]+).
Example 55
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-O--(4,5-
dihydro-1H-imidazol-2-yl-methyl)-tyrosine
The methyl ester was prepared by reaction of N-(toluene-4-sulfonyl)-
L-prolyl)-L-O-cyanomethyl-tyrosine methyl ester with ethylene diamine (in
refluxing methanol under Argon overnight). The product was purified by
flash column chromatography (silica, 5% methanol in chloroform) to afford
the methyl ester as a white foam. The title compound was prepared using the
procedure described in Method 6 and was isolated as a solid, mp = 147-
149°C.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): 8 = 7.74 (d, 2H, J = 8.4Hz); 7.61
(d, 1 H, J = 5.7Hz); 7.40 (d, 2H, J = 8.2Hz); 6.97 (d, 2H, J = 8.6Hz); 6.75
(m,
2H); 6.51 (br s, 1 H); 4.49 (s, 2H); 3.96 (dd, 1 H; 3 = 2.5, 6Hz); 3.82 (dd, 1
H,
J = 5.1, 9.6Hz); 3.59 (t, 1H, J = 9.7Hz); 3.21 (t, 1H, J =9.6Hz); 3.04 (m,
4H);
2.39 (s, 3H); 1.70 {m, 1H); 1.35 (m, 3H).
IR (KBr, cm''): 3400, 2900, 1650, 1610, 1510, 1400, 1315, 1245,
1150, 1075, 1040, 700, 600, 550.
Mass Spectroscopy: (ESI, m/e (%)) 515 (100, (M+H)+)
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Example 56
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(3-propyl-ureido)-phenylalanine
The methyl ester was prepared by the reaction of N (toluene-4-
sulfonyl)-L-prolyl-L-(4-amino)-phenylalanine methyl ester with propyl
isocyanate (triethylamine, toluene, reflux 1.25 hours). The crude product
was purified by flash chromatography (silica, 1:1 EtOAc:hexane) to give the
methyl ester as a white foam. The title compound was prepared using the
procedure described in Method 6.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): 8 = 12.78 (br s, 1 H); 8.29 (s, 1 H);
7.99 {d, 1 H, J = 8.1 Hz); 7.68 (d, 2H, J = 8.1 Hz); 7.39 (d, 2H, J = 8.1 Hz);
7.26 (d, 2H, J = 8.SHz); 7.06 (d, 2H, J = B.SHz); 6.04 (t, 1H, J = 5.7Hz};
4.39
{m, 1 H); 4.1 (dd, 1 H, J = 2.9, 8.2Hz); 3.3 (m, 1 H); 3.09 (m, 1 H); 2.99 (m,
3H); 2.87 (dd, 1H, J = 8.34, 13.$3Hz); 2.38 (s,3H); 1.49-1.64 (m, 3H); 1.40
(m, 3H); 0.84 (t, 3H, J = 7.46Hz).
IR (KBr, cm''): 3590, 3350, 3220, 3050, 2960, 2930, 2860, 1750,
1660, 1630, 1590, 1560, 1540, 1345, 1320, 1160, 660, 590.
Mass Spectroscopy: ( (+) FAB, m/e (%)) 539 (40, [M + Na]+); 517
(20 [M+H]+}.
Example 57
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-benzylamino)phenylalanine
The title compound was prepared from the product of Example 58
using the procedure described in Method 6.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHZ): 8 7.72 (d, 2H, J = 8.3 Hz); 7.54 (d,
1 H, J = 5.49 Hz); 7.4 (d, 2H, J = 7.9 Hz); 7.27 (m, 4H); 717 (m, 1 H); 6.72
(d,
2H, J = 8.~6 Hz); 6.34 {d, 2H, J = 8.56 Hz); 5.91 (t, 1 H, J = 6.15 Hz); 4.17
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(d, 2H, J = 6.15 Hz); 3.91 (dd, 1 H, J = 2.74, 8.89 Hz); 3.75 (m, 1 H); 2.95
(m,
2H); 2.88 (m, 2H); 2.38 (s, 3H); 1.66 (m, 1H); 1.18-1.42 (m, 3H).
1R (KBr, cm'') 3400, 1655, 1620, 1525, 1410, 1340, 1160, 670.
MS ((+) FAB, m/e (%)) 528 ( I 00 [M + Li]+); 522 ( 15 [M+H]+)
S Anal. Calc'd for CZ8H3°N305S Li 1.5 H20; C, 60.64; H, 5.90; N,
7.57.
Found: C, 60.60; H, 5.61; N, 7.45.
Example 58
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-
L-(4-benzylamino)phenylalanine methyl ester
The title compound was prepared by reductive amination of N-
(toluene-4-sulfonyl)-L-prolyl-L-4-aminophenylalanine methyl ester with
benzaldehyde (acetic acid, sodium triacetoxyborohydride, methylene
chloride, stirred at room temperature overnight). The crude product was
purified by flash chromatography (silica, 1:1 EtOAc:hexane) to afford the
title compound as a white solid, mp = 53-56 ° C (0.501 g, 41 %).
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): 8 8.09 (d, 1 H, J = 7.90 Hz); 7.68 (d,
2H, J = 8.34 Hz); 7.39 (d, 2H, J = 7.90 Hz); 7.28 (m. 4H); 717 (m, 1 H); 6.87
(d, 2H, J = 8.34 Hz); 6.46 (d, 2H, J = 8.56 Hz); 6.12 (t, 1 H, J = 6.15 Hz);
4.38 (m, 1H); 4.20 (d, 2H, J = 5.71 Hz); 4.07 (m, 1 H); 3.58 (s, 3H); 3.27 (m,
1H); 3.06 (m, 1H); 2.82 (m, 2H); 2.39 (s, 3H); 1.53 (m, 3H); 1.36 (m, 1H).
IR (KBr, cm'') 3400, 2950, 1745, 1675, 1610, 1525, 1450, 1350,
1210, 1100, 680, 595, 550.
MS ((+) FAB, m/e (%)) 536 (70 [M+H]+)
Anal. Calc'd for C,9H3;N3OSS: C, 65.03: H, 6.21; N, 7.84.
Found: C, 64.57; H, 6.10; N, 7.58.
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Example 59
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-
(4-chlorobenzylamino)-phenylalanine
The methyl ester was prepared by reductive amination of N (toluene-
4-sulfonyl)-L-prolyl-L-(4-amino)-phenylalanine methyl ester with 4-
chlorobenzaldehyde (acetic acid, sodium triacetoxyborohydride, methylene
chloride, stirred at room temperature for 2.5 hours). The crude product was
purified by flash chromatography (silica, 1:1 EtOAc:hexane) to afford the
methyl ester as a white solid. The title compound was prepared using the
procedure described in Method 6.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz): 8 = 7.72 (d, 2H, J = 8.3Hz); 7.53 (d,
1H, J = 5.7Hz); 7.4 (d, 2H, J = 7.9Hz); 7.3 (s, 4H); 6.72 (d, 2H, J = 8.3Hz);
I 5 6.31 (d, 2H, J = 8.5Hz); 6.0 (m, 1 H); 4.16 {d, 2H, J = 6.1 Hz); 3.92 (dd,
1 H, J
= 9, 2.85Hz); 3.75 (m, 1H); 2.91 (m, 4H); 2.38 (s, 3H); 1.65 (m, 1H); 1.35
(m, 1 H); 1.18 (m, 2H).
IR (KBr, cm'): 3400, 1620, 1525, 1410, 1340, 670.
Mass Spectroscopy: ( (+) FAB, m/e (%)) 578 (95, [M + Na]+); 562
(50, [M+ Li]+); 556 (40 [M+H]+),
Example 60
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl)-L-
(4-chloromethanesulfonylamino)-phenylalanine
N (Toluene-4-sulfonyl)-L-prolyl-L-(4-amino)-phenylalanine methyl
ester (I.OOg, 2.2 mmol) was dissolved in methylene chloride (2.5 mL) and
cooled to -78°C. With stirring, chloromethane sulfonyl chloride (0.34g,
2.2
mmol) was added followed by dropwise addition of pyridine (182 mL, 2.2
mmol). The clear solution became yellow, then orange, then finally red upon
the chloromethanesulfonyl chloride addition. The solution was allowed to
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warm to room temperature and was stirred for 16 hr. The reaction solution
was transferred to a 250 mL separatory funnel with CHzCl2 (50 mL) and
extracted with 1N HCl (50 rnL x 2), brine (50 mL x 2), and water (50 mL).
The organic phase was dried (MgS04) and the solvent removed to give a red
solid which was flash chromatographed (Si02, 3:1 CH,C1~:CH3CN) to yield a
white solid. The title compound was prepared using the procedure described
in Method 6 and was isolated as a solid, mp = 163-170°C.
NMR data was as follows:
'H NMR (DMSO-db, 400 Mhz): b = 8.08 (d, IH, J = 7.7Hz); 7.71
(dd, 4H, J = 14.7Hz, 8.3Hz); 7.51 (d, 1H, J = 5.9Hz); 7.41-7.39 (m, 4H);
6.84-6.82 (m, 2H); 6.78-6.72 (m, 4H); 6.68 (d, 2H, J = 7.5Hz); 4.40 (q, 1H, J
= 7.7Hz); 4.18 (s, 3H); 4.19-4.08 (m, 1H); 3.95-3.89 (m, 2H); 3.60 (s, 2H);
3.11-3.04 (m, 2H); 2.95-2.83 (m, 3H); 2.39 (s, 3H); 1.78-1.65 (m, 1H); 1.61-
1.56 (m, 3H); 1.42-1.37 (m, 4H).
IR (KBr, cm-1): 3400, broad; 2960; 2850; 1749; 1650; 1510; 1450;
1400; 1350; 1300; 1260; 1160; 1100; 1000; 840; 800; 650; 600; 550.
Mass Spectroscopy: (-) ESI: 542.1 (M-H); 483.8; 442.2; 341.6;
273.8; 202.1; 144.9.
Example 61
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
thiaprolyl-L-4-(aminobenzoyl)phenylalanine methyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): b = 7.89-7.85 (m, 3H), 7.74 (d, 2H), 7.60-7.47 (m,
5H), 7.29 (d, 2H), 7.21 (d, 2H), 7.02 (d, 1 H), 4.89 (m, 1 H), 4.56 (d, 1 H),
4.37
(d, 1H), 3.85 (s, 1H), 3.75 (s, 3H), 3.12 (m, 2H), 2.44 (s, 3H), 1.18 (s, 3H),
1.08 (s, 3H).
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'3C NMR (CDC13): 8 = 171.8, 169.1, 166.3, 145.4, 137.6, 135.8,
133.3, 132.7, 132.5, 130.7, 130.6, 129.4, 128.7, 127.6, 120.7, 74.1, 55.2,
53.9, 53.1, 51.1, 38.4, 29.8, 24.6, 22.3.
.5 Example 62
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-
thiaprolyl-L-4-(benzamido)phenylalanine
The title compound was prepared from the product of Example 61
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CDCl3): b = 8.43 (s, 1H), 7.84 (d, 2H), 7.72 (d, 2H), 7.61
(d, 2H), 7.49-7.20 (m, 7H), 4.93 (q, 1 H), 4.55 (d, 1 H), 4.37, (d, 1 H), 3.92
(s,
1H), 3.15 (m, 2H), 2.42 (s, 3H), 1.19 (s, 3H), 1.08 (s, 3H).
'3C NMR (CDCl3): 8 = 177.62, 174.35, 169.69, 145.53, 137.73,
135.51, 133.19, 132.44, 132.39, 130.79, 130.63, 129.23, 128.63, 127.82,
121.1, 73.94, 55.13, 53.97, 51.08, 38.02, 29.79, 24.62, 22.24.
Example 63
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
thiaprolyl-L-tyrosine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): 8 = 7.73 (d, 2H), 7.33 (d, 2H), 7.08-7.04 {m, 3H),
6.73 (d, 2H), 6.51 (s, 1H), 4.88 (e, 1H), 4.54 (d, 2H), 4.33 (d, 2H),1.28 (t,
3H), 4.23 (q, 2H), 3.83 (s, 1H), 3.14-2.93 (m, 2H), 2.43 (s, 3H), 1.03 (s,
3H),
1.02 (s, 3H).
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'3C NMR (CDC13): b = 171.56, 169.44, 155.87, 145.40, 133.16,
131.16, 130.59, 128.66, 127.97, 116.00, 74.01, 62.35, 55.11, 54.16, 51.04,
38.33, 29.79, 24.26, 22.24, 14.67.
Example 64
Synthesis of
N (Toluene-4-Sulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L-tyrosine
The title compound was prepared from the product of Example 63
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CDCl3): 8 = 7.96 (d, 1H), 7.49 (d, 2H), 7.17 (d, 2H), 6.91
(d, 2H), 6.49 (d, 2H), 4.40 (m, 1H), 4.38-4.27 (dd, 2H), 3.83 (s, 1H), 2.95-
2.70 (m, 2H), 2.23 (s, 3H), 0.94 (s, 3H), 0.92 (s, 3H).
Example 65
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-4-[(pyridin-4-yl)methylamino]phenyianine
The title compound was prepared from the product of Example 66
using the procedure described in Method 6.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHZ): 8 = 8.42 (d, 2H, J = 5.93Hz); 7.72
(d, 2H, J = 8.34Hz); 7.53 (d, 1H, J = 5.71Hz); 7.39 (d, 2H, J = 7.90Hz); 7.28
{d, 2H, J = 5.93Hz); 6.74 (d, 2H, J = 8.56Hz); 6.31 (d, 2H, J = 8.34Hz); 6.07
(t, 1 H, J = 6.26Hz); 4.22 (d, 2H, J = 6.15Hz); 3.91 (dd, 1 H, J = 2.85,
8.78Hz); 3.77 (dd, 1H J = 5.38, 9.99 Hz); 2.95 {m, 2H); 2.89 (m, 2H); 2.38
(s, 3H); 1.66 (m, 1H); 1.19-1.38 (m, 3H).
IR (KBr, cm') 3400, 1650, 1610, 1520, 1420, 1340, 1155.
MS ((-) ESI, m/e {%)) 521 (100[M-H]')
Anal. Calc'd for C,,HZ9N405S Li 2.5 H,O: C, 56.47; H, 5.97; N, 9.7.6.
Found: C, 56.32; H, 5.61; N, 9.64.
*rB
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Example 66
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4
[(pyridin-4-yl)methylamino]phenylanine methyl ester
Substituting 4-pyridinecarboxaldehyde for benzaldehyde, and
following the method for the preparation of Example 58, gave the title
compound.
NMR data was as follows:
'H NMR (DMSO-db, 400 MHz) 8 = 8.43 (d, 2H, J = 6.1 SHz); 8.10 (d,
1H, J = 7.90 Hz); 7.67 (d, 2H, J = 8.12Hz); 7.39 (d, {2H, J=7.90Hz); 7.28 (d,
2H, J = 6. i SHz); 6.89 (d, 2H, J = 8.56 Hz); 6.43 (d, 2H, J = 8.56); 6.26 (t,
1H, J = 6.26 Hz); 4.38 (m, IH); 4.26 (d, 2H), J = 6.37Hz); 4.06 (m, IH); 3.58
(s, 3H); 3.28 (m, 1H); 3.06 (m, IH); 2.84 (m, 2H); 2.39 (s, 3H), 1.51 (m,
3H); 1.34 (m, 1H).
IR (KBr, cm-') 3400, 2930, 1745, 1675, 1615, 1600, 1520, 1345,
1 i60, 1090, 1000, 770, 720, 600, 550.
MS ((+) FAB, m/e (%)) 537 (75[m+H]+).
Example 67
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L-4
(pyridin-3-carboxamido)phenylalanine methyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): 8 = 9.01 (d, IH), 8.74 (m, 1H), 8.23 (m, 2H), 7.73
(d, 2H), 7.59 (d, 2H), 7.43 (m, 1H), 7.33 (d, 2H), 7.27 (m, IH), 7.20 (d, 2H),
4.84 (m, 1H), 4.40-4.32 (dd, 2H), 3.86 (s, 1H), 3.74 (s, 3H), 3.15-3.10 (m,
2H), 2.44 (s, 3H), 1.19 (s, 3H), 1.05 (s, 3H).
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'3C NMR (CDC13): 8 = 171.8, 169.2, 164.3, 152.9, 148.5, 145.4,
137.2, 136.1, 133.3, 133.2, 131.3, 130.8, 130.6, 128.6, 124.3, 121.0, 74.1,
55.1, 54.1, 53.0, 51.1, 38.2, 29.8, 24.6, 22.2.
S Example 68
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
thiaprolyl-L-4-(pyridine-3-carboxamido)phenylalanine
The title compound was prepared from the product of Example 67
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CD3)ZSO): b = 9.10 (s, 1H), 8.73 (d, 1H), 8.33 (d, 1H), 7.78
(d, 2H), 7.73 (d, 1 H), 7.62 (d, 2H), 7.53 (m, 1 H), 7.41 (d, 2H), 7.20 {d,
2H),
4.51 (dd, 2H), 4.08-4.03 (m, 2H), 3.00 (m, 2H), 2.39 (s, 3H), 1.19 (s, 3H),
1.10 (s, 3H).
'3C NMR (CD3)ZSO): 8 = 173.1, 167.1, 164.1, 152.3, 149.1, 144.3,
137.0, 135.8, 135.2, 134.2, 131.1, 130.3, 130.2, 128.3, 123.8, 120.1, 73.0,
55.9, 54.9, 50.6, 37.5, 29.9, 25.0, 21.4.
Example 69
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L
4-[(pyridin-3-ylmethyl)amino]phenylalanine
Substituting 3-pyridinecarboxaldehyde for benzaldehyde, and
following the method for the preparation of Example 58, gave the title
compound.
NMR data was as follows:
'H NMR {DMSO-d6,400 MHz) 8 = 8.51 (s, 1H); 8.38 (m, 1H); 7.72
{d, 2H, J=8.13Hz); 7.68 (m, 1 H); 7.54 (d, 1 H, J=5.71 Hz); 7.40 (d, 2H,
J=7.90Hz); 7.28 (m, 1H); 6.75 (d, 2H, J=8.56 Hz); 6.36 (d, 2H, J=8.56Hz);
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5.98 (t, 1H, J=5.27, 9.88Hz); 2.96 (m, 2H); 2.89 (m, 2H); 2.39 (s, 3H); 1.65
(m, 1H); 1.16-1.43 (m, 3H).
IR (KBr, cm'') 3400, 1670, 1620, 1520, 1400, 1345, 1160, 675, 600.
MS ((+) FAB, m/e (%)) 523 (25[M+H]+); 529 (100[M + Li]+)
Anal. Calc'd for C2,HZ9N405S Li 2.5 HzO: C,56.47; H, 5.97; N, 9.76.
Found: C, 56.62; H, 5.57; N, 9.69.
Example 70
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-
thiaprolyl-L-4-nitrophenylalanine methyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): 8 = 8.14 (d, 2H), 7.74 (d, 2H), 7.41 (d, 2H), 7.35
(d, 2H), 7.10 (d, 1 H), 4.94 (m, 1 H), 4.58 (d, 1 H), 4.33 (d, 1 H), 3.82 (s,
1 H),
3.76 {s, 3H), 3.25 (m, 2H), 2.44 (s, 3H), I.l 1 (s, 3H), 1.02 (s, 3H).
'3C NMR (CDCl3): b = 171.3, 169.3, 147.7, 145.6, 144.6, 133.0,
131.0, 130.6, 128.6, 124.2, 74.0, 54.9, 53.6, 53.3, 51.1, 38.7, 29.8, 24.5,
22.3.
Example 71
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-nitrophenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): 8 = 8.16 (d, 2H), 7.71 (d, 2H), 7.48 (d, 1H), 7.40-
7.31 (m, 4H), 4.87 (q, 1 H), 4.24 (q, 2H), 4.07 (m, 1 H), 3.44-3.35 (m, 2H),
3.23-3.05 (m, 2H), 2.44 (s, 3H), 2.04 (m, 1H), 1.62-1.44 (m, 3H), 1.30 (t,
3H).
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'3C NMR (CDC13): b = 171.5, 170.9, 147.6, 145.1, 144.7, 133.2,
130.9, 130.6, 128.4, 124.2, 62.7, 62.6, 53.7, 50.8, 38.4, 30.1, 25.0, 22.2,
14.7.
Example 72
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4-(2
methoxybenzamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): 8 = 9.81 (s, 1 H), 8.27(d, 1 H), 7.71 (d, 2H), 7.60
{d, 2H), 7.49 (t, 1H), 7.37-7.28 (m, 3H), 7.18-7.10 (m, 3H), 7.03 (d, 1H),
4.82 (m, 2H), 4.22 (q, 2H), 4.10-4.04 (m, 4H), 3.40 (m, IH), 3.30-3.02 (m,
3H), 2.42 (s, 3H), 2.03 (m, 1H), 1.53 (m, 3H), 1.30 (t, 3H).
'3C NMR (CDC13): b = 171.4, 163.7, 157.8, 144.9, 137.9, 133.9,
133.6, 133.1 ,132.4, 130.5, 128.5, 122.3, 120.9, 112.1, 62.9, 62.3, 56.8,
54.1,
50.3, 38.0, 30.5, 24.9, 22.2, 14.7.
Example 73
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
thiaprolyl-L-(4-nitro)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): b = 8.12 (d, 2H), 7.73 (d, 2H), 7.42 {d, 2H), 7.34
(d, 2H), 7.12 (d, 1 H), 4.92 (m, 1 H), 4.57 (d, 1 H), 4.3 3 (d, 1 H), 4.20 (q,
2H),
3.83 (s, 1H), 3.33-3.15 (m, 2H), 2.43 (s, 3H), 1.25 (t, 3H), 1.11 (s, 3H),
1.01
(s, 3H).
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'3C NMR (CDC13): b = 170.8, 169.3, 147.6, 145.5, 144.7, 133.0,
131.1, 130.6, 128.6, 124.2, 74.0, 62.6, 54.9, 53.6, 51.0, 38.8, 29.8, 24.5,
22.2,
14.7.
Example 74
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4
(2-bromobenzamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCI~): b = 7.78 (s, 1H), 7.70 (d, 2H), 7.65-7.55 (m, 4H),
7.42-7.29 (m, SH), 7.16 (d, 2H), 4.81 (m, 1 H), 4.23 (q, ZH), 4.06 (m, 1 H),
3.43-3.00 (m, 4H), 2.43 (s, 3H), 2.08-1.99 (m, 1H), 1.60-1.45 (m, 3H), 1.30
(t, 3H).
'3C NMR (CDCl3): 8 = 171.5, 171.3, 166.1, 144.9, 138.3, 137.1,
134.1, 133.5, 133.3, 132.3, 130.6, 130.5, 130.3, 128.5, 128.3, 120.6, 119.8,
62.9, 62.3, 51.0, 50.3, 38.0, 30.4, 24.9, 22.2, 14.7.
Example 75
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-diraethyl)
thiamorphyl-L-4-aminophenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 38.
NMR data was as follows:
'H NMR (CDC13): b = 7.74 (d, 2H), 7.33 (d, 2H), 6.96 (m, 3H), 6.59
(d, 2H), 4.81 (m, 1 H), 4.53 (d, 1 H), 4.38 (d, 1 H), 4.19 (q, 2H), 3.85 (s, 1
H),
3.61 (br s, 2H), 2.98 (m, 2H), 2.44 (s, 3H), 1.26 (t, 3H), 1.17 (s, 3H), 1.10
(s,
3H).
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'3C NMR (CDC13): b = 171.5, 168.8, 125.9, 145.3, 133.4, 130.9,
130.5, 128.7, 126.3, 115.8, 74.1, 62.1, 55.3, 54.1, 38.2, 29.8, 24.6, 22.2,
14.7.
Example 76
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-4-acetamidophenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 39.
NMR data was as follows:
'H NMR (CDCl3): b = 7.69 (d, 2H), 7.52 (s, 1H), 7.42 (d, 2H), 7.38-
7.30 (m, 3H), 7.09 (d, 2H), 4.78 (m, 1H}, 4.21 (q, 2H}, 4.04 (m, 1H), 3.38
(m, 1H), 3.25-2.97 (m, 3H), 2.43 (s, 3H), 2.13 (s, 3H), 2.02 (m, 1H), 1.58-
1.44 (m, 3H), 1.28 (t, 3H).
'3C NMR (CDCl3): 8 = 171.4, 169.0, 144.9, 137.6, 133.4, 132.4,
130.6, 130.4, 128.4, 120.4, 62.9, 62.3, 54.0, 50.3, 37.9, 30.4, 25.2, 24.9,
22.2,
14.7.
Example 77
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-4-(2-methoxybenzamido)phenylalanine
The title compound was prepared from the product of Example 72
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CD30D): b = 7.82 (d, 1 H), 7.67 (d, 1 H), 7.51 (d, 2H), 7.41
(d, 2H), 7.31 (t, 1 H), 7.19 (d, 2H), 7.05 (d, 2H), 6.97 (d, 1 H), 6.89 (t, 1
H),
4.49 (m, 1H}, 3.91 (m, 1H), 3.78 (s, 3H), 3.23-2.83 (m, 3H), 2.21 (S, 3H),
1.68-1.28 (m, 4H).
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Example 78
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
thiaprolyl-L-4-acetamidophenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 39.
NMR data was as follows:
'H NMR (CDCl3): b = 7.73 (d, 2H), 7.50 (brs, 1H), 7.42 (d, 2H), 7.33
(d, 2H), 7.15 (d, 2H), 7.02 (d, 1 H), 4. 84 (m, 1 H), 4.53 (d, 1 H), 4.3 5 (d,
1 H),
4.18 (q, 2H), 3.84 (s, 1H), 3.08 (m, 2H), 2.43 (s, 3H), 2.15 (s, 3H), 1.25 (t,
3H), 1.13 (s, 3H), 1.06 (s, 3H).
'3C NMR (CDCl3): 8 = 171.3, 169.1, 163.0, 145.4, 137.6, 133.3,
132.4, 130.6, 128.7, 120.4, 112.0, 74.1, 62.3, 55.1, 54.0, 41.1, 38.4, 29.8,
25.2, 24.5, 22.2, 14.7.
Example 79
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-
thiaprolyl-L-4-acetamidophenylalanine
The title compound was prepared from the product of Example 78
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CDCl3): 8 = 8.19 (s, 1 H), 7.73 (d, 2H), 7.40 (d, 2H), 7.32
(d, 2H), 7.23 (d, 1H), 7.15 (d, 2H), 4.86 (m, 1H), 4.55 (d, 1H), 4.36 (d, 1H),
3.92 (s, 1H), 3.10 (m, 2H), 2.42 (s, 3H), 2.10 (s, 3H), 1.19 (s, 3H), 1.07 (s,
3H).
*rB
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Example 80
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-4-acetamidophenylalanine Isopropyl Ester
The title compound was prepared following the procedure outlined
for the preparation of Example 39.
NMR data was as follows:
'H NMR (CDC13): b = 7.70 (d, 2H), 7.42 (d, 2H), 7.33 (d, 2H), 7.10
(d, 2H), 5.11-5.03 (m, 1 H), 4.80-4.73 (m, 1 H), 4.06 (dd, 1 H), 3.41-3.36 (m,
1 H), 3.20 (dd, 1 H), 3.12-3.09 (m, 1 H), 3.02 (dd, 1 H), 2.44 (s, 3H), 2.15
(s,
3H), 2.05-2.01 (m, 1H), 1.58-1.44 (m, 3H), 1.27 (d, 3H).
'3C NMR (CDC13): b = 170.7, 170.2, 168.3, 144.3,.136.9, 132.9,
131.9, 129.9, 129.9, 127.8, 119.7, 69.5, 62.3, 53.5, 49.7, 37.3, 29.8, 24.6,
24.2, 21.7, 21.7, 21.6.
Example 81
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
L-4-(isonicotinamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): b = 8.75 (d, 2H), 8.27 (s, 1 H), 7.74-7.68 (m, 4H),
7.59 (d, 2H), 7.33 (d, 2H), 7.22 (d, 2H), 7.08 (d, 1 H), 4.86 (m, 1 H), 4.54
(d,
1 H), 4.34 (d, 1 H), 4.18 (q, 2H), 3.84 (s, 1 H), 3.18-3.03 (m, 2H), 2.43 (s,
3H),
1.26 (t, 3H), 1.26 (t, 3H), 1.15 (s, 3H), 1.04 (s, 3H).
'3C NMR (CDCI3): 8 = 171.3, 169.1, 164.3, 151.2, 145.4, 142.6,
137.0, 133.4, 133.2, 130.8, 130.6, 128.6, 121.6, 121.0, 74.1, 62.4, 55.1,
54.0,
51.1, 3 8.4, 29.8, 24.6, 22.2, 14.7.
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Example 82
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4
(isonicotinamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): b = 8.76 (d, 2H), 8.34 (s, 1 H), 7.72 (d, 2H), 7.68
(d, 2H), 7.59 (d, 2H), 7.40 (d, 1 H), 7.32 (d, 2H), 7.16 {d, 2H), 4.80 (m, 1
H),
4.21 (q, 2H), 4.04 (m, 1H), 3.40 (m, 1H), 3.28-3.02 (m, 3H), 2.43 (s, 3H),
2.03 (m, 1H), 1.60-1.42 (m, 3H), 1.29 (t, 3H).
"C NMR (CDC13): 8 = 171.4, 171.3, 164.3, 151.2, 145.0, 142.6,
137.0, 133.5, 133.4, 130.6, 130.6, 128.4, 121.6, 121.0, 62.9, 62.3, 54.1,
52.3,
38.0, 30.3, 24.9, 22.2, 14.7.
Example 83
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-(~-toluene-4-sulfonyl)histidine methyl ester
L-Prolyl-L-histidine methyl ester was treated with CH3SO~C1 and
Et3N in CH~C12 to give the title compound.
NMR data was as follows:
'H NMR (CDC13): 8 = 7.93 (s, 1H), 7.81 (d, 2H). 7.77 (d, 1H), 7.73
(d, 2H), 7.34 (d, 4H), 7.09 (s, 1 H), 4.82 (m, 1 H), 4.76-4.09 (dd, 1 H), 3.68
(s,
3H), 3.51-3.44 (m, 1H), 3.18-3.08 (m, 3H), 2.45 (s, 3H), 2.41 (s, 3H), 2.10-
2.05 (m, 1H), i.68-1.53 (m, 3H).
'3C NMR (CDCl3): 8 = 171.1, 171.0, 146.2, 144.2, 140.0, 136.5,
134.8, 133.2, 130.4, 129.9, 127.8, 127.4, 114.8, 62.2, 52.5, 51.9, 49.5, 30.1,
29.9, 24.3, 21.7, 21.6.
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Example 84
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-thiaprolyl-L
4-(nicotinamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): 8 = 9.06 (s, 1H), 8.76 (d, 1H), 8.22 {d, 1H), 8.14
(s, 1H), 7.73 (d, 2H), 7.59 (d, 2H), 7.45 (t, 1H), 7.33 (d, 2H), 7.23 (d, 2H),
7.18 (d, 1 H), 4.86 (m, 1 H), 4.52 (d, 1 H), 4.34 (d, 1 H), 4.20 (q, 2H), 3.06-
2.20 (m, 2H), 2.43 (s, 3H), 1.27 (t, 3H), 1.20 (s, 3H), 1.08 (s, 3H).
'3C NMR (CDC13): b = 171.3, 169.1, 152.9, 148.4, 145.4, 137.1,
136.1, 133.4, 130.8, 130.6, 128.7, 124.3, 120.9, 74.1, 62.3, 55.1, 54.0, 51.1,
38.7, 29.8, 24.6, 22.3, 14.7.
Example 85
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-(O-methyl)tyrosine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): 8 = 7.71 (d, 2H), 7.32 (m, 3H), 7.06 (d, 2H), 6.80
(d, 2H), 4.77 (m, 1 H), 4.21 (q, 1 H), 4.18-4.06 (m, 1 H), 3.77 (s, 3 H), 3.37-
3.34 (m, 1H), 3.20-3.09 (m, 2H), 3.00 (dd, 3H), 2.43 (s, 3H), 1.55-1.46 (m,
3H), 1.28 (t, 3H).
''C NMR (CDCl3): b = 171.5, 171.3, 159.2, 144.9, 133.6, 130.9,
130.5, 128.7, 128.4, 114.4, 62.8, 62.2, 55.8, 54.1, 50.2, 37.7, 30.3, 24.8,
22.3,
14.7.
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Example 86
Synthesis of
N (a-Toluenesulfonyl)-L-prolyl
L-4-(isonicotinamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): 8 = 8.72 (d, 2H), 8.45 (s, 1H), 7.69 (d, 2H), 7.55
(d, 2H), 7.45-7.30 (m, SH), 7.13 (d, 2H), 6.99 (d, 1H), 4.78 (m, 1H), 4.28 (s,
2H), 4.18 (q, 2H), 4.03 (m, 1H), 3.23-2.98 (m, 4H), 2.10-1.62 (m, 4H), 1.27
(t, 3H).
'3C NMR (CDCl3): 8 = 171.6, 174.5, 164.4, 151.1, 142.6, 137.1,
133.3, 131.4, 130.6, 129.5, 129.4, 129.0, 121.6, 12I.I, 62.9, 62.4, 57.3,
53.9,
50.1, 37.9, 31.0, 25.5, 14.7.
Example 87
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-4-(2-bromobenzamido)phenylalanine
The title compound was prepared from the product of Example 74
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CD30D): 8 = 8.01 (brd, 1 H), 7.75-7.58 (m, 4H), 7.50-7.33
(m, 4H), 7.24 (d, 2H), 4.70 (m, IH), 3.50-3.00 (m, SH), 2.43 (s, 3H), 1.85-
1.48 (m, 4H).
Example 88
Synthesis of
N (a-Toluenesulfonyl)-L-prolyl-
L-4-(isonicotinamido)phenylalanine
The title compound was prepared from the product of Example 86
using the procedure described in Method 7.
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NMR data was as follows:
'H NMR (Dz0): 8 = 8.45 (d, 2H), 7.50 (d, 2H), 7.26 {d, 2H), 7.18 (s,
SH), 7.05 (d, 2H), 4.29 (m, 1H), 4.10 (m, 2H), 3.83 (m, 1H), 3.20-2.97 (m,
3H), 2.76 {m, 1H), 1.94 (m, 1H), 1.70-1.43 (m, 3H).
Example 89
Synthesis of
N (Toluene-4-sulfonyl)-L-(1,1-dioxo)thiamorpholyl-
L-4-(2-bromobenzamido))phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36 and the procedure described by Larsson et
al., Acta Chemica Scan., 1994, 48, 522.
NMR data was as follows:
'H NMR (CDC13): 8 = 8.19 (d, 1H), 7.70 (d, 1H), 7.61-7.50 (m, SH),
7.36-7.23 (m, 3H), 7.09-7.04 (m, 2H), 6.84 (d, 1/2H), 6.74 (d, 1/2H), 5.10 (d,
1 /2H), 4.97 (d, 1 /2H), 4.81 (m, 1 /2H), 4.66 (m, 1 /2H) 4.14 (m, 3H), 3.87
(m,
1H), 3.23-3.74 (m, 6H), 2.41 (s, 3H), 1.22 (t, 3H).
"C NMR (CDC13): b = 170.9, 170.7, 166.0, 165.8; 165.1, 164.8,
145.6, 145.5, 137.9, 136.9, 136.7, 135.1, 135.0, 133.4, 132.0, 131.9, 131.5,
130.7, 130.6, 130.0, 130.1, 129.5, 129.4, 127.6, 127.4, 127.2, 120.7, 120.2,
61.8, 55.9, 55.8, 53.5, 53.3, 49.3, 49.2, 48.7, 48.6, 41.8, 41.7, 36.8, 36.3,
21.5, 13.9.
Example 90
Synthesis of
N (Toluene-4-sulfonyl)-L-(1,1-dioxo)thiamorpholyl
L-4-(2-bromobenzamido)phenylalanine
The title compound was prepared from the product of Example 89
using the procedure described in Method 7.
NMR data was as follows:
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'H NMR (CD30D): 8 = 8.12 (d, 0.5H), 7.97 (d, 0.5H), 7.76-7.59 (m,
5.5H), 7.60-7.18 (m, 7H), 5.12 (m, 1 H), 4.64 (m, 1 H), 4.07 (m, 1 H), 3.62
(m,
1H), 3.35 (m, 6H), 2.40 (s, 3H), 2.32-2.14 (m, 3H).
'3C NMR (CD30D): 8 = 174.1, 169.7, 169.6, 169.2, 169.1, 146.4,
146.2, 140.3, 140.3, 138.7, 138.6, 137.4, 137.2, 134.9, 134.6, 134.3, 132.5,
131.6, 131.5, 131.2, 131.1, 130.0, 129.9, 128.9, 128.7, 128.6, 122.1, 121.8,
120.6, 56.9, 56.8, 54.9, 54.5, 50.8, 50.3, 40.8, 10.4, 37.8, 37.4, 21.6.
Example 91
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)
L-4-{isonicotinamido)phenylalanine
The title compound was prepared from the product of Example 81
using the procedure described in Method 7.
Physical data was as follows:
M.p. >200°C;
MS(FAB+) 583(M+H).
Example 92
Synthesis of
N (a-Toluenesulfonyl)-L-prolyl-L-4
{2-bromobenzamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): b = 7.37 (s, 1H, 7.68-7.30 (m, 11H), 7.16 (d, 2H),
6.93 (d, 1 H), 4.83 (m, 1 H, 4.32 (s, 2H, 4.23 (q, 2H), 4.10 (m, 1 H), 3.27-
3.03
(m, 5H), 2.10-1.62 (m, 4H), 1.29 (t, 3H).
'3C NMR (CDC13): 8 = 166.7, 161.2, 133.4, 132.2, 129.2, 128.1,
127.3, 126.4, 125.7, 125.4, 125.6, 124.5, 124.1, 123.4, 115.7, 114.9, 57.8,
57.3, 55.9, 52.4, 48.7, 45.1, 32.8. 26.0, 20.4, 9.6.
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Example 93
Synthesis of
N (Toluene-4-sulfonyl)-L-(S,S-dimethyl)-
thiaprolyl-L-tyrosine Isoropyl Ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): b = 7.73 (d, 2H), 7.33 (d, 2H), 7.10 (br d, 1H),
7.08 (d, 2H), 6.75 (d, 2H), 5.06 (m, 1 H), 4.86 (m, 1 H), 4.54 (d, 1 H), 4.33
(d,
1 H), 3.84 (s, 1 H), 3.09-2.97 (m, 2H), 2.43 (s, 3H), 1.23 (dd, 6H), 1.05 (s,
3H), 1.02 (s, 3H).
'3C NMR (CDCl3): 8 = 170.6, 169.0, 155.3, 144.9, 132.6, 130.6,
130.0, 128.1, 127.2, 115.4, 73.3, 69.6, 65.8, 54.4, 53.6, 50.3, 37.6, 29.0,
23.5,
21.5, 21.5.
Example 94
Synthesis of
N (Toluene-4-sulfonyl)-L-(5,5-dimethyl)-
thiaprolyl-L-tyrosine tent-Butyl Ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): 8 = 7.75 (d, 2H), 7.34 (d, 2H), 7.11 (d, 2H), 7.07
(d, 1H), 6.76 (d, 2H), 6.62 (br s, 1H), 4.79 (m, 1H), 4.56 (d, 1H), 4.35 (d,
1H), 3.84 (s, 1H), 3.07-2.96 (m, 2H), 2.45 (s, 3H), 1.46 (s, 9H), 1.05 (s,
3H),
1.03 (s, 3H).
'3C NMR (CDCl3): 8 = 170.1, 168.8, 155.3, 144.9, 132.6, 130.7,
130Ø 128.1, 127.6, 115.3, 82.7, 73.3, 54.5, 54.1, 50.4, 37.8, 29.1, 27.8,
23.5,
21.5.
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Example 95
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-tyrosine tert-Butyl Ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): b = 7.62 (d, 2H), 7.35 (d, 1H), 7.25 (d, 2H), 6.96
(d, 2H), 6.71 (d, 2H), 4.65 (m, 1 H), 4.07 (m, 1 H), 3.30 (m, 1 H), 3.15-2.83
(m, 3H), 2.35 (s, 3H), 1.84 (m, 1H), 1.52-1.34 (m, 13H).
'3C NMR (CDC13): b = 171.5, 170.0, 155.8, 144.5, 132.8, 130.4,
130.0, 127.8 ,127.1, 115.3, 82.5, 62.2, 54.1, 49.5, 37.0, 29.8, 27.7, 23.9,
21.3,
14Ø
Example 96
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl-L-4
(2-trifluoromethytbenzamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): b = 7.81 (s, 1H), 7.50 - 7.38 {m, 8H), 7.38 (d, 1H),
7.33 (d, 2H), 7.14 (d, 2H), 4.80 (m, 1H), 4.22 (q, 2H), 4.03 (m, 1H), 3.37 (m,
1H), 3.30 - 3.20 (m, 1H), 3.14 - 2.98 (m, 2H), 2.43 (s, 3H), 1.95 (m, 1H),
1.57 - 1.40 (m, 3H), 1.30 (t, 3H).
''H NMR (CDC13): 8 = 171.2, 170.9, 165.8, 144.4, 136.6, 135.7,
132.9, 132.8, 132.2, 130.7, 130.2, 130.0, 130.0, 128.5, 127.9, 126.5, 120.2,
62.1, 61.7, 53.3, 49.6, 37.2, 29.7. 24.0, 21.4, 13.9.
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Example 97
Synthesis of
N (Toluene-4-sulfonyl)-L-proly!
L-4-(2-methylbenzamido)phenylalanine ethyl ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDCl3): 8 = 7.72 (d, 2H), 7.58-7.43 (m, 4H), 7.40-7.20 (m,
7H), 7.15 (d, 2H), 4.82 (m, 1 H), 4.24 (g, 2H), 4.08 {m, 1 H), 3.43 (m, 1 H),
3.29-3.03 (m, 3H), 2.50 (s, 3H), 2.44 (s, 3H), 2.05 (m, 1H), 1.55 {m, 3H),
1.31 (t, 3H).
'3C NMR (CDCl3): b = 170.9, 170.9, 144.4, 137.0, 136.5, 133.0,
132.4, 131.3, 130.3, 130.1, 130.0, 127.9, 126.6, 125.9, 119.8, 62.2, 61.6,
53.4, 49.6, 37.3, 29.6, 24.2, 21.4, 19.7, 14Ø
Example 98
Synthesis of
N (Toluene-4-sulfonyl)-L-prolyl
L-4-(2-trifluoromethylbenzamido)phenylalanine
The title compound was prepared from the product of Example 96
using the procedure described in Method 7.
NMR data was as follows:
'H NMR (CDCl3): 8 = 8.28 (s, 1H), 7.68-7.40 (m, 9H), 7.33 (d, 2H),
7.17 (d, 2H), 4.84 (m, 1 H), 4.05 (m, 1 H), 3.44-3.04 (m, 4H), 2.43 (s, 3H),
1.90 (m, 1H), 1.49 (m, 3H).
Example 99
Synthesis of
N (4-Fluorobenzenesulfonyl)-L-thiaprolyl-L-tyrosine tent-Butyl Ester
The title compound was prepared following the procedure outlined
for the preparation of Example 36.
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NMR data was as follows:
'H NMR (CDCl3): b = 7.87 (m, 2H), 7.23 (m, 3H), 7.03 (d, 2H), 6.73
(d, 2H), 6.29 (s, 1 H), 4.68 (m, 1 H), 4.56 (m, 2H), 4.10 (d, 1 H), 3.29-2.93
(m,
3H), 2.61 (m, 1H), 1.49 (s, 9H).
'3C NMR (CDCl3): 8 = 169.9, 168.2, 155.4, 132.5, 130.9, 130.7,
127.3, 117.1, 116.8, 115.4, 82.9, 65.0, 54.1, S 1.5, 36.8, 33.3, 27.9.
Example 100
Synthesis of
N (4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl)-
thiaprolyl-L-tyrosine tert-Butyl Ester
The title compound was prepared following the procedure described
for the preparation of Example 36.
NMR data was as follows:
'H NMR (CDC13): 8 = 7.90 (m, 2H}, 7.23 (m, 2H), 7.09 {d, 2H), 6.98
(d, 1 H), 6.76 (d, 2H), 4.77 (m, 1 H), 4.55 (d, 1 H), 4.3 7 (d, 1 H), 3.83 (s,
1 H),
3.01 (m, 1H), 1.44 (s, 9H), 1.10 (s, 3H),1.05 (s, 3H).
''C NMR (CDC13): 8 = 170.1, 168.6, 155.3, 132.0, 131.0, 130.8,
127.7, 116.9, 116.6, 115.3, 82.8, 73.4, 54.6, 54.0, 50.4, 37.8, 29.1, 27.8,
23.5.
Example 203
Synthesis of
N-(Toluene-4-sulfonyl)-L-prolyl-L-4-(4-vitro-phenoxycarbonyloxy)
phenylalanine ethyl ester
In a nitrogen atmosphere, Tos-Pro-Tyr ethyl ester was dissolved in
CH~Ch at RT, the flask cooled to 0°C and 4-
nitrophenylchloroformate added
in one portion. Next, Et3N was added dropwise slowly, stirred at 0°C
for 30
minutes and then at RT for 30 minutes. The reaction mixture was again
cooled to 0°C and N,N-dimethylethanolamine was added dropwise. The ice
bath was taken away after 10 minutes and the reaction mixture stirred at RT
for 3 hours. The reaction mixture was diluted with Et,O and washed with
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10% K,C03 until all yellow color disappeared. Flash chromatography (Si02)
eluting with 40% EtOAc/hex yielded a white foam in 30% yield.
Example 204
Synthesis of N-(Toluene-4-sulfonyl)-L-(5,5-diraethyl)
thiaprolyl-4-(2-bromobenzaraido)-D-phenylalanine t-butyl ester
The starting (2R)-3-(4-amino-phenyl)-2-{[(4R)-5,5-dimethyl-3-
(toluene-4-sulfonyl)-thiazolidine-4-carbonyl]-amino}-propionic acid
tert-butyl ester (230 mg), 2-bromobenzoic acid ( 100 mg), and
4-methylmorpholine (0.19 mL) were dissolved in DMF (5 mL) at 0°C in an
ice bath. BOP (229 mg) was added to the solution. The ice bath was
removed after 10 minutes. The reaction mixture was stirred at room
temperature for 18 hours. Ethyl acetate (20 mL) was added. The mixture
was washed with citric acid (5%, 20 mL, 2x) and saturated NaHC03 solution
(20 mL, 2x), then washed with brine. The solution was dried with MgS04.
The solvent was evaporated in vacuo and the residue was purified using
HPLC (reverse phase, 65-95% CH30H/water). The retention time was 13
minutes. The desired product was freeze dried to give 181 mg of a white
solid. MP:92-93°C
Example 205
Synthesis of N-(Toluene-4-sulfonyl)-L-(5,5-diraethyl)thiaprolyl-4
(2-bromobenzamido)-L-phenylalanine t-butyl ester
The starting (2S)-3-(4-amino-phenyl)-2-{ [(4R)-5,5-dimethyl-3-
(toluene-4-sulfonyl)-thiazolidine-4-carbonyl]-amino}-propionic acid
tert-butyl ester (230 mg), 2-bromobenzoic acid ( 100 mg), and
4-methylmorpholine (0.19 mL) were dissolved in DMF (5 mL) at 0°C in an
ice bath. BOP (229 mg) was added to the solution. The ice bath was
removed after 10 minutes. The reaction mixture was stirred at room
temperature for 18 hours. Ethyl acetate (20 mL) was added. The mixture
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was washed with citric acid (5%, 20 mL, 2x) and saturated NaHC03 solution
(20 mL, 2x), then washed with brine. The solution was dried with MgS04.
The solvent was evaporated in vacuo and the residue was purified using
HPLC (reverse phase, 65-95% CH30H/water). The retention time was 13.5
minutes. The desired product was freeze dried to give 27 mg of the title
compound as a white solid. MP: 92-93°C
Example 207
Synthesis of N-(Toluene-4-sulfonyl)-L-prolyl-4-(1-H,
2-oxo-3-methyltetrahydropyrimidin-1-yl)-L-phenylalanine
The title compound was prepared from the corresponding t-butyl
ester using the procedure described in Method 11.
NMR data was as follows:
'H NMR (CDC13): & = 7.73 (d, 2H), 7.58 (d, 1H), 7.34 (d, 2H), 7.21
(d, 2H), 7.17 (d, 2H), 4.79 (q, 1 H), 4.15 - 4.1 I (m, 1 H), 3.68 - 3.63 (m,
2H),
3.48 - 3.39 (m, 3H), 3.27 (dd, 1 H), 3.17 (dd, 1 H), 3. I 5 - 3.07 (m, 1 H),
2.99
(s, 3H), 2.43 (s, 3H), 2.16 -2.08 (m, 2H), 2.00 - 1.98 {m, 1H).
"C NMR (CDC13):8 = 173.4, 172.2, 164.2, 156.4, 144.4, 142.5, 134.1, 133.0,
130.2, 130.0, 127.9, 126.2, 62.1, 53.4, 49.5, 48.9, 47.9, 36.5, 35.9, 30.2,
24.2,
22.0, 21.4.
Example 212
N-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl) thiaprolyl-L-3-chloro-4
hydroxyphenylalanine Isopropyl Ester
The compound was prepared following the procedure described for
the preparation of Example 36 employing N-(4-fluorobenzenesulfonyl)-L-
(5,5-dimethyl, thiaproline (prepared according to Method Band 3-chloro-L-
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tyrosine isopropyl ester (prepared from commercially available 3-chloro-L-
tryosine and isopropyl alcohol in the presence of anhydrous HCl).
MS:(-) ESI [M-H)-557
HPLC: 96.8%
Example 214
Synthesis of
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methoxyphenyl)-L-phenylalanine
The title compound was prepared from the corresponding t-butyl
ester using the procedure described in Method 11.
NMR data was as follows:
'H NMR (CD30D):8 = 7.70 (m, 2H), 7.36 (m, 4H), 7:22 (m, 4H),
6.98 (m, 2H), 4.75 (m, 1H), 4.10 (m, 1H), 3.71 (s, 3H), 3.29 (m, 2H), 3.11
(m, 2H), 2.39 (s, 3H), 1.75 (m, IH), 1.53 (m, 3H).
''C NMR (CD30D):8 = 174.4, 174.2, 158.1, 145.9, 138.9, 136.7,
135.1, 131.2, 130.9, 130.8, 130.2, 129.9, 129.1, 122.0, 112.6, 63.3, 55.9,
54.6, 50.5, 37.9, 31.5, 25.2, 21.4.
Example 215
Synthesis of
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2-methoxyphenyl)-L-phenylalanine
The title compound was prepared from the corresponding t-butyl
ester using the procedure described in Method 11.
NMR data was as follows:
'H NMR (CDCl3): b 8.41 (d, 1 H), 8.21 (s, 1 H), 8.03 (d, 1 H), 7.98 (s,
1 H), 7.74 (d, 2H), 7.39 (d, 1 H), 7.33 (d, 2H), 4.72-4.68 (m, 1 H), 4.17-4.13
(m, 1 H). 3.54-3.34 (m, 3H), 3.20-3.12 (m, 1 H), 2.82 (s, 6H)m 2.43 (s, 3H),
2.09-2.04 (m, 1H), 1.79-1.59 (m, 3H).
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'3C NMR (CDC13):8 = 173.7, 171.8, 154.5, 147.2, 144.4, 137.8,
135.5, 133.2, 130.1, 127.9, 126.4, 62.2, 53.0, 49.5, 38.5, 36.0, 30.3, 24.4,
21.4.
Example 216
N-(Toluene-4-sulfonyl)-L-prolyl-4-(N,N-dimethylsulfamyl)
L-phenylalanine
The title compound was prepared from the corresponding tert-butyl
ester using the procedure described in Method 11.
NMR data was as follows:
'H NMR (CDCl3):8 = 7.71 (d, 2H), 7.58 (d, 1H), 7.49 (bs, 1H), 7.34
(d, 2H), 7.16 (d, 2H), 7.12 (d, 2H), 4.90 - 4.83 (m, 1 H), 4.13 - 4.09 (m, 1
H),
3.46 - 3.40 (m, 1 H), 3.28 (dd, 1 H), 3.15 - 3.04 (m, 2H), 2.77 (s, 6H), 2.43
(s,
3H), 1.93 - 1.88 (m, 1H), 1.54 - 1.45 (m, 3H).
'3C NMR (CDCl3):8 = 173.7, 172.0, 144.7, 136.6, 132.6, 132.1,
130.4, 130.2, 127.9, 120.3, 62.2, 53.3, 49.7, 38Ø 36.8, 29.9, 24.1, 21.4.
Example 217
Synthesis of
N-(Toluene-4-sulfonyl)-L-prolyl-(NI,N1,N2-trimethylsulfamyl)-
L-phenylalanine
The title compound was prepared from the corresponding ten-butyl
ester using the procedure described in Method 11.
NMR data was as follows:
'H NMR (CDCl3):8 = 7.71 (d, 2H), 7.49 (d, 1 H), 7.35 (d, 4H), 7.24
(d, 2H), 4.89 - 4.82 (m, 1 H), 4.10 - 4.07 (m, 1 H), 3.41 - 3.31 (m, 2H), 3.22
(s, 3H), 3.16 - 3.06 (m, 2H), 2.76 (s, 6H), 2.44 (s, 3H), 1.98 - 1.95 (m, 1H),
1.55 - 1.38 (m, 3H).
'3C NMR (CDCl3):8 = 173.7, 171.8, 144.6, 141.7, 135.1, 132.7,
130.2, 103.1, 127.9, 126.6, 62.2, 53.2, 49.6, 39.4, 38.2, 36.7, 29.6, 24.1,
21.5.
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Example 220
Synthesis of
N-[4-(3,3-Dimethyl-ureido)-benzenesulfonyl)-L-prolyl-(4-hydroxy)
L-phenylalanine isopropyl ester
The reaction vessel was flushed with NZ, Pd/C was added to the flask
and wet with iso-PrOH. Next, the nitro compound was added to the flask in
iso- PrOH and a balloon filled with HZ was attached to the flask. The NZ was
vacuumed out, HZ added, evacuated, and Hz added again. The reaction was
complete in 2 hours; diluted with iso-PrOH and filtered through Celite and
evaporated to dryness. Flash chromatography (SiO,) diluting with 1%
MeOH/CHZCI, yielded a white solid in 62% yield of the lower Rf from the
separation.
Example 225
Synthesis of
N-(Toluene-4-sulfonyl)-L-prolyl-4-{N,N-dimethylaminocarbonylmethyl)-
L-phenylalanine
The title compound was prepared from the product of Example 233
using the procedure described in Method I 1.
NMR data was as follows:
'H NMR (CD30D): b = 8.04 (d, I H), 7.70 (d, 2H), 7.40 (d, 2H), 7.20
(m, 4H), 4.99 (bs, 1 H), 4.70 (m, 1 H), 4.10 (m, 1 H), 3.69 (s, 2H), 3.40 (m,
1H), 3.20 (m, 1H), 3.04 (m, 2H), 3.01 (s, 3H), 2.92 (s, 3H), 2.4I (s, 3H),
1.80
(m, 1 H), 1.62 (m, 3H).
''C NMR (CD30D):8 = 174.2, 174.1, 1.73.9, 145.8, 136.8, 135.0,
131.2, 131.0, 130.0, 129.1, 63.2, 54.7, 54.6, 50.5, 41.0, 38.3, 37.7, 35.9,
31.6,
25.2, 21.5.
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Example 226
Synthesis of
N-(1-Methylimidazol-4-sulfonyl)-L-prolyl-(4-hydroxyl)-L-phenylalanine
isopropyl ester
The compound was prepared by BOP coupling of
1-methylimidazole-4-sulfonyl-Pro with Tyr-isopropyl ester. The crude
product was purified by flash chromatography {silica, 95:3:2
EtOAc:EtOH:Et3N) to afford a white solid, m.p. = 159-162°C.
(2.03g, 65%).
MS((+)ESI, m/z {%)) 465 (100 [M+H]+).
Example 227
Synthesis of
N-(Toluene-4-sulfonyl)-L-prolyl-4-(2,4,5-trioxo-3-phenyltetrahydro
imidazol-1-yl)-L-phenylalanine isopropyl ester
The compound was prepared by treatment of N-(toluene-4-sulfonyl)-
L-prolyl-4-[(phenyl ureido)-tetrahydroimidazol-1-yl]-L-phenylalanine
isopropyl ester with oxalyl chloride in methylene chloride. The crude
product was purified by flash chromatography (silica, 3:2 Hex:EtOAc) to
afford a white solid. (0.490g, 49%).
MS((+)ESI, m/z (%)) 647 (15 [M+H]+); 664 (100[M+NH4]+.
Example 229
N-(4-Fluorobenzenesulfonyl)-L-(5,5-dimethyl) thiaprolyl-L-3-chloro
4-~t-butoxy-phenylalanine tart-Butyl Ester
The compound was prepared according to the procedure described in
Example 36. The required 3-chloro-O-tart-butyl-L-tyrosine tart-butyl ester
was prepared from 3-chloro-L-tyrosine and iso-butylene in the presence of
one equivalent of sulfuric acid and used without further purification.
MS-(+)ESI [M+H]+ 629 [M+NH4]+ 646.
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Example 231
N-[2-(N-2,10-camphorsultamyl) acetyl]
-L-3-chloro-4-hydroxyphenylalanine Isopropyl Ester
S The above compound was prepared by the following steps:
Step Preparation of N-(tert-butoxycarbonylmethyl)
camphorsultam
Camphorsultam (3g, 14 mmole) in DMF (25 mol) was mixed with
NaH (1.1 equiv.) for 15 minutes, then tert-butyl bromoacetate (2.5 ml) was
added and the reaction mixture was stirred overnight. The solvent was
evaporated to dryness and the residue was partitioned between aq-citric
acid and EtoAc. The organic layer was washed with sat. aq. NaHC43, and
water, dried and evaporated to give the ester. (3.5 g)
1 S te~2_: Preparation of N-(carboxymethyl) camphorsultam
The ester (3.3g) was converted to the acid according to Method 17.
MS (-)ESI [M-H]- 272
~p~ Preparation of N-[2-(N-2,10-camphorsultamyl) acetyl]-
L-3-chloro-4-hydroxyphenylalanine Isopropyl Ester
The titled compound was prepared by amide bond formation
according to a method similar to Example 52.
MS:(+)ESI [M+H]+513
HPLC : 99.4
Example 236
Synthesis of
N-(4-Fluorobenzenesulfonyl)-L-prolyl-4-hydroxy-L-phenylalanine
t-butyl ester
The compound was prepared using the procedure described in
Method 12. The crude product was purified by flash column
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chromatography (silica, hexane:EtOAc 1:1 ) to give the title compound as a
white foam (580 mg, 64%).
MS (+)ESI, [M+H]+ @ rn/z 493 [M+NH4]+ @ rn/z 510
Example 237
Synthesis of
N-(4-Fluorobenzenesulfonyl)-L-prolyl-4-hydroxy-L-phenylalanine
ispropyl ester
The compound was prepared using the procedure described in
Method 12. The crude product was purified by flash column
chromatography (silica, hexane:EtOAc 1:1) (380 mg, 43%).
MS (+)ESI, [M+H]+ @ tn/z 479 (100%) [M+NH,]+ @ m/z 496
Example 238
Synthesis of
N-(1-Methylimidazol-4-sulfonyl)-L-prolyl-4-hydroxy-L-phenylalanine
t-butyl ester
The compound was prepared by BOP coupling of
1-methylimidazole-4-sulfonyl-Pro with Tyr-t-butyl ester. The crude product
was purified by flash chromatography (silica. 95:3:2 EtOAc: EtOH:Et3N) to
afford a white solid. (1.21 g, 66%)
MS ((+)ESI, m/z (%)) 479 (100 [M+H]+)
Example 239
Synthesis of N-(Pyridine-3-sulfonyl)-L-prolyl-4-hydroxy-L-
phenylalanine t-butyl ester
The compound was prepared by BOP coupling of
3-pyridinesulfonyl-Pro with Tyr-t-butyl ester. The crude product was
purified by flash chromatography (silica, 95:3:2 EtOAc: EtOH:Et3N) to
afford a pale orange foam. (0.930g, 95%)
MS ((+)ESI, m/z (%)) 476 (100 [M+H]+)
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Example 240
Synthesis of N-(Toluene-4-sulfonyl)-L-prolyl-4-(methanesulfonamido)
L-phenylalanine
To a solution of 3-(4-methanesulfonylamino-phenyl)-
2-{[1-(toluene-4-sulfonyl)-pyrrolidine-2-carbonyl]-amino}-propionic acid
methyl ester (0.250 g, 0.477 mmol) in THF (I .0 mL) was added IN LiOH
(955 mL, 0.955 mmol) and the clear solution was stirred at 25°C, under
nitrogen, for 16 h. The reaction solution was then diluted with additional
water (SOmL) and washed with diethyl ether (25 mL). The aqueous layer was
IO lyophiiized to afford a fluffy white solid (0.216 g, 87%).
MS (-ESI): 508 [M - I]-
Example 242
Synthesis of
15 N-(Toluene-4-sulfonyl)-L-prolyl-4-(2,4,5-trioxo-3-(3-chlorophenyl)-tetra
hydroimidazol-1-yl)-L-phenylalanine benzyl ester
The compound was prepared by treatment of N-(toluene-4-sulfonyl)-
L-prolyl-4-[(3-chlorophenyl ureido)-tetrahydroimidazol-I-yl]-L-
phenylalanine isopropyl ester with oxalyl chloride in methylene chloride.
20 The crude product was purified by flash chromatography (silica, 3:2 Hex:
EtOAc) to afford a white solid. (0.41 Og, 50%).
MS ((+)ESI, m/z (%) 746 (100[M+H]+) (746/748 1C1)
Certain of the above exemplified compounds as well as certain other
25 compounds which can be prepared by the methods described above include
those set forth in Tables IA and IB above.
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Example 244
In vitro Assay For Determining Binding of
Candidate Compounds to VLA-4
An in vitro assay was used to assess binding of candidate compounds
to a4(3, integrin. Compounds which bind in this assay can be used to assess
VCAM-1 levels in biological samples by conventional assays (e.g.,
competitive binding assays). This assay is sensitive to ICso values as low as
about 1 nM.
The activity of a4~i, integrin was measured by the interaction of
soluble VCAM-1 with Jurkat cells (e.g., American Type Culture Collection
Nos. TIB 152, TIB I53, and CRL 8163), a human T-cell line which
expresses high levels of a4~i, integrin. VCAM-1 interacts with the cell
surface in an a4~i, integrin-dependent fashion (Yednock, et al. J. Biol.
Chem.,
1995, X0:28740).
Recombinant soluble VCAM-1 was expressed as a chimeric fusion
protein containing the seven extracellular domains of VCAM-1 on the N-
terminus and the human IgG, heavy chain constant region on the C-terminus.
The VCAM-1 fusion protein was made and purified by the manner described
by Yednock, supra.
Jurkat cells were grown in RPMI 1640 supplemented with 10% fetal
bovine serum, penicillin, streptomycin and glutamine as described by
Yednock, supra.
Jurkat cells were incubated with 1.5 mM MnCI, and 5 ~g/mL 15/7
antibody for 30 minutes on ice. Mn+2 activates the receptor to enhance
ligand binding, and 1 S/7 is a monoclonal antibody that recognizes an
activated/ligand occupied conformation of a4(3, integrin and locks the
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molecule into this conformation thereby stabilizing the VCAM-1/x4[3,
integrin interaction. Yednock, et al., supra. Antibodies similar to the 15/7
antibody have been prepared by other investigators {Luque, et al, 1996, J.
Biol. Chem. x:11067) and may be used in this assay.
Cells were then incubated for 30 minutes at room temperature with
candidate compounds, in various concentrations ranging from 66 pM to 0.01
~M using a standard 5-point serial dilution. 15 ~L soluble recombinant
VCAM-1 fusion protein was then added to Jurkat cells and incubated for 30
minutes on ice. (Yednock et al., supra.).
Cells were then washed two times and resuspended in PE-conjugated
goat F(ab')z anti-mouse IgG Fc (Immunotech, Westbrook, ME) at 1:200 and
incubated on ice, in the dark, for 30 minutes. Cells were washed twice and
analyzed with a standard fluorescence activated cell sorter ("FACS") analysis
as described in Yednock, et al., supra.
Compounds having an ICSO of less than about 15~.M possess a
suitable binding affinity to a4y.
When tested in this assay, each of the compounds in Examples 1-243
has an ICSO of 15 ~cM or less.
Example 245
In vitro Saturation Assay For Determining Binding of
Candidate Compounds to a4~3,
The following describes an in vitro assay to determine the plasma
levels needed for a compound to be active in the Experimental Autoimmune
Encephalomyelitis ("EAE") model, described in the next example, or in other
in vivo models.
*rB
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Log-growth Jurkat cells are washed and resuspended in normal
animal plasma containing 20 wg/ml of the 15/7 antibody (described in the
above example).
The Jurkat cells are diluted two-fold into either normal plasma
samples containing known candidate compound amounts in various
concentrations ranging from 66 ~,M to 0.01 ~M, using a standard 12 point
serial dilution for a standard curve, or into plasma samples obtained from the
peripheral blood of candidate compound-treated animals.
Cells are then incubated for 30 minutes at room temperature, washed
twice with phosphate-buffered saline ("PBS") containing 2% fetal bovine
serum and 1mM each of calcium chloride and magnesium chloride (assay
medium) to remove unbound 15/7 antibody.
The cells are then exposed to phycoerythrin-conjugated goat F(ab')2
anti-mouse IgG Fc (Immunotech, Westbrook, ME), which has been adsorbed
for any non-specific cross-reactivity by co-incubation with 5% serum from
the animal species being studied, at 1:200 and incubated in the dark at
4°C
for 30 minutes.
Cells are washed twice with assay medium and resuspended in the
same. They are then analyzed with a standard fluorescence activated cell
sorter ("FACS") analysis as described in Yednock et al. J. Biol. Chem., 1995,
270:28740.
The data is then graphed as fluorescence versus dose, e.g., in a
normal dose-response fashion. The dose levels that result in the upper
plateau of the curve represent the levels needed to obtain efficacy in an in
vivo model.
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This assay may also be used to determine the plasma levels needed to
saturate the binding sites of other integrins, such as the a9(3, integrin,
which
is the integrin most closely related a4~i, (Palmer et al, 1993, J. Cell Bio.,
123:1289). Such binding is predictive of in vivo utility for inflammatory
conditions mediated by a9~3, integrin, including by way of example, airway
hyper-responsiveness and occlusion that occurs with chronic asthma, smooth
muscle cell proliferation in atherosclerosis, vascular occlusion following
angioplasty, fibrosis and glomerular scarring as a result of renal disease,
aortic stenosis, hypertrophy of synovial membranes in rheumatoid arthritis,
and inflammation and scarring that occur with the progression of ulcerative
colitis and Crohn's disease.
Accordingly, the above-described assay may be performed with a
human colon carcinoma cell line, SW 480 (ATTC #CCL228) transfected
1 S with cDNA encoding a9 integrin (Yokosaki et al., I 994, J. Biol. Chem.,
269:26691 ), in place of the Jurkat cells, to measure the binding of the a9(3,
integrin. As a control, SW 480 cells which express other a and ~3, subunits
may be used.
Using a conventional oral formulation, compounds of this invention
would be active in this model.
Example 246
In vivo Evaluation
The standard multiple sclerosis model, Experimental Autoimmune
(or Allergic) Encephalomyelitis ("EAE"), was used to determine the effect of
candidate compounds to reduce motor impairment in rats or guinea pigs.
Reduction in motor impairment is based on blocking adhesion between
leukocytes and the endothelium and correlates with anti-inflammatory
activity in the candidate compound. This model has been previously
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described by Keszthelyi et al., Neurology, 1996, 47:1053-1059, and
measures the delay of onset of disease.
Brains and spinal cords of adult Hartley guinea pigs were
homogenized in an equal volume of phosphate-buffered saline. An equal
volume of Freund's complete adjuvant (100 mg mycobacterium tuberculosis
plus 10 ml Freund's incomplete adjuvant) was added to the homogenate. The
mixture was emulsified by circulating it repeatedly through a 20 ml syringe
with a peristaltic pump for about 20 minutes.
Female Lewis rats (2-3 months old, 170-220 g) or Hartley guinea
pigs (20 day old, 180-200 g) were anesthetized with isoflurane and three
injections of the emulsion, 0.1 ml each, were made in each flank. Motor
impairment onset is seen in approximately 9 days.
Candidate compound treatment began on Day 8, just before onset of
symptoms. Compounds were administered subcutaneously ("SC"), orally
("PO") or intraperitoneally ("IP"). Doses were given in a range of l0mg/kg
to 200 mg/kg, bid, for five days, with typical dosing of 10 to 100 mg/kg SC,
10 to 50 mg/kg PO, and 10 to 100 mg/kg IP.
Antibody GGS/3 against a4~3, integrin (Keszthelyi et al., Neurology,
1996, 47:1053-1059), which delays the onset of symptoms, was used as a
positive control and was injected subcutaneously at 3 mg/kg on Day 8 and
11.
Body weight and motor impairment were measured daily. Motor
impairment was rated with the following clinical score:
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0 no change
1 tail weakness or paralysis
2 hindlimb weakness
3 hindlimb paralysis
4 moribund or dead
A candidate compound was considered active if it delayed the onset
of symptoms, e.g., produced clinical scores no greater than 2 or slowed body
weight loss as compared to the control.
When tested in this in vivo assay, the compounds of Examples 10 and
27 were active.
Example 247
In vivo Evaluation - Asthma
Inflammatory conditions mediated by a4~3, integrin include, for
example, airway hyper-responsiveness and occlusion that occurs with
chronic asthma. The following describes an asthma model which can be
used to study the in vivo effects of the compounds of this invention for use
in
treating asthma.
Following the procedures described by Abraham et al, J. Cl in. Invest.
X3:776-787 (1994) and Abraham et al, Am J. Respir Crit Care Med. x:696-
703 (1997), both of which are incorporated by reference in their entirety,
compounds of this invention are formulated into an aerosol and administered
to sheep which are hypersensitive to Ascaris suum antigen. Compounds
which decrease the early antigen-induced bronchial response and/or block the
late-phase airway response, e.g., have a protective effect against antigen-
induced late responses and airway hyper-responsiveness ("AHR"), are
considered to be active in this model.
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Allergic sheep which are shown to develop both early and late
bronchial responses to inhaled Ascaris swum antigen are used to study the
airway effects of the candidate compounds. Following topical anesthesia of
the nasal passages with 2% lidocaine, a balloon catheter is advanced through
one nostril into the lower esophagus. The animals are then intubated with a
cuffed endotracheal tube through the other nostril with a flexible fiberoptic
bronchoscope as a guide.
Pleural pressure is estimated according to Abraham (1994). Aerosols
(see formulation below) are generated using a disposable medical nebulizer
that provides an aerosol with a mass median aerodynamic diameter of 3.2 ~m
as determined with an Andersen cascade impactor. The nebulizer is
connected to a dosimeter system consisting of a solenoid valve and a source
of compressed air (20 psi). The output of the nebulizer is directed into a
plastic T-piece, one end of which is connected to the inspiratory port of a
piston respirator. The solenoid valve is activated far 1 second at the
beginning of the inspiratory cycle of the respirator. Aerosols are delivered
at
VT of 500 ml and a rate of 20 breaths/minute. A 0.5% sodium bicarbonate
solution only is used as a control.
zo
To assess bronchial responsiveness, cumulative concentration-
response curves to carbachol can be generated according to Abraham (1994).
Bronchial biopsies can be taken prior to and following the initiation of
treatment and 24 hours after antigen challenge. Bronchial biopsies can be
preformed according to Abraham (1994).
An in vitro adhesion study of alveolar macrophages can also be
performed according to Abraham (1994), and a percentage of adherent cells
is calculated.
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Aerosol Formt lax tion
A solution of the candidate compound in 0.5% sodium
bicarbonate/saline (w/v) at a concentration of 30.0 mg/mL is prepared using
the following procedure:
~. P,r~enaration of 100.0 mL of 0~,5%
Sodium Bicarbonate / Saline Stock Solution
Ingredient Gram / 100.0 mL Final Concentration
Sodium Bicarbonate0.5 g 0.5%
Saline q.s. ad 100.0 q.s. ad 100%
mL
Procedure:
1. Add O.Sg sodium bicarbonate into a 100 mL voh.~metric flask.
2. Add approximately 90.0 mL saline and sonicate until
dissolved.
3. Q.S. to 100.0 mL with saline and mix thoroughly.
B. Preparation of 30.0 mg/mL Candidate ~ompoun~l: 10.0~~L
zo
Ingredient Gram / 10.0 mL Final Concentration
Candidate Compound 0.300 g 30.0 mg/mL
0.5% Sodium Bicarbonateq.s. ad 10.0 q.s ad 100%
/ mL
Saline Stock Solution
Procedure:
1. Add 0.300 g of the candidate compound into a 10.0 mL
vohzmetric flask.
2. Add approximately 9.7 mL of 0.5% sodium bicarbonate /
saline stock solution.
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3. Sonicate until the candidate compound is completely
dissolved.
4. Q.S. to 10.0 mL with 0.5% sodium bicarbonate / saline stock
solution and mix thoroughly.
Using a conventional oral formulation, compounds of this invention
would be active in this model.
*rB
