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TITLE
ISOXAZOLINE AND ISOXAZOLE DERIVATIVES AS INTEGRIN RECEPTOR ANTAGONISTS
FIELD OF THE INVENTION
This invention relates to novel heterocycles which
are useful as antagonists of the av03 and related
integrin receptors, to pharmaceutical compositions
containing such compounds, processes for preparing such
compounds, and to methods of using these compounds,
alone or in combination with other therapeutic agents,
for the inhibition of cell adhesion and the treatment of
angiogenic disorders, inflammation, bone degradation,
tumors, metastases, thrombosis, and other cell
aggregation-related conditions.
BACKGROUND OF THE INVENTION
Angiogenesis or neovascularization is critical for
normal physiological processes such as embryonic
development and wound repair (Fol}anan and Shing, J.
Biol. Chem. 1992, 267:10931-10934; D'Amore and Thompson,
Ann. Rev. Physiol. 1987, 49:453-464). However,
angiogenesis occurs pathologically, for example, in
ocular neovascularization (leading to diabetic
retinopathy, neovascular glaucoma, retinal vein
occlusion and blindness), in rheumatoid arthitis and in
solid tumors (Folkman and Shing, J. Biol. Chem., 1992,
267:10931-10934; Blood and Zetter, Biochim. Biophys.
Acta., 1990, 1032:118-128).
Tumor dissemination, or metastasis, involves
= several distinct and complementary components, including
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the penetration and transversion of tumor cells through
basement membranes and the establishment of self-
sustaining tumor foci in diverse organ systems. To this
end, the development and proliferation of new blood f
vessels, or angiogenesis, is critical to tumor survival.
Without neovascularization, tumor cells lack the
nourishment to-divide and will not be able to leave the
primary tumor site (Folkman and Shing, J. Biol. Chem.,
1992, 267:10931-10934).
Inhibition of angiogenesis in animal models of
cancer has been shown to result in tumor growth
suppression and prevention of metastatic growth (Herblin
et al., Exp. Opin. Ther. Patents. 1994, 1-14). Many
angiogenic inhibitors have been directed toward blocking
initial cytokine-dependent induction of new vessel
growth, e.g. antibodies to endothelial cell growth
factors. However, these approaches are problematic
because tumor and inflammatory cells can secrete
multiple activators of angiogenesis (Brooks et al.,
Cell, 1994, 79:1157-1164). Therefore, a more general
approach that would allow inhibition of angiogenesis due
to a variety of stimuli would be of benefit.
The integrin av03 is preferentially expressed on
angiogenic blood vessels in chick and man (Brooks et
al., Science, 1994, 264:569-571; Enenstein and Kramer,
J. invest. Dermatol., 1994, 103:381-386). Integrin avP3
is the most promiscuous member of the integrin family,
allowing endothelial cells to interact with a wide
variety of extracellular matrix components (Hynes, Cell,
1992, 69:11-25). These adhesive interactions are
considered to be critical for angiogenesis since
vascular cells must ultimately be capable of invading
virtually all tissues.
While integrin avP3 promotes adhesive events
important for angiogenesis, this receptor also transmits
signals from the extracellular environment to the =
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intracellular compartment (Leavesley et al., J. Cell
Biol., 1993, 121:163-170, 1993). For example, the
interaction between the avP3 integrin and extracellular
matrix components promotes a calcium signal required for
cell motility.
During endothelium injury, the basement membrane
zones of blood vessels express several adhesive
proteins, including but notlimited to von Willebrand
factor, fibronectin, and fibrin. Additionally, several
members of the integrin family of adhesion receptors are
expressed on the surface of endothelial, smooth muscle
and on other circulating cells. Among these integrins
is av/03, the endothelial cell, fibroblast, and smooth
muscle cell receptor for adhesive proteins including von
Willebrand factor, fibrinogen (fibrin), vitronectin,
thrombospondin, and osteopontin. These integrins
initiate a calcium-dependent signaling pathway that can
lead to endothelial cell, smooth muscle cell migration
and, therefore, may play a fundamental role in vascular
cell biology.
Recently, an antibody to the avP3 integrin has been
developed that inhibits the interaction of this integrin
with agonists such as vitronectin (Brooks et al.,
Science, 1994, 264:569-571). Application of this
antibody has been shown to disrupt ongoing angiogenesis
on the chick chorioallantoic membrane (CAM), leading to
rapid regression of histologically distinct human tumor
transplanted onto the CAM (Brooks et al., Cell, 1994,
79:1157-1164). In this model, antagonists of the avP3
integrin induced apoptosis of the proliferating
angiogenic vascular cells, leaving pre-existing
quiescent blood vessels unaffected. Thus, avP3 integrin
antagonists have been shown to inhibit angiogenesis.
= Based on this property, therapeutic utility of such
agents is expected in human diseases such as cancer,
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rheumatoid arthritis and ocular vasculopathies (Folkman
and Shing, J. Biol. Chem., 1992, 267:10931-10934).
Increasing numbers of other cell surface receptors
have been identified which bind to extracellular matrix
ligands or other cell adhesion ligands thereby mediating
cell=cell and cell-matrix adhesion processes. These
receptors belong to a gene superfamily called integrins
and are composed of heterodimeric transmembrane
glycoproteins containing a- and O-subunits. Integrin
subfamilies contain a common P-subunit combined with
different a-subunits to form adhesion receptors with
unique specificity. The genes for eight distinct ~-
subunits have been cloned and sequenced to date.
Two members of the pi subfamily, a4/P1 and a5/pi have
been implicated in various inflammatory processes.
Antibodies to a4 prevent adhesion of lymphocytes to
synovial endothelial cells in vitro, a process which may
be of importance in rheumatoid arthritis (VanDinther-
Janssen et al., J. Immunol., 1991, 147:4207).
Additional studies with monoclonal anti-a4 antibodies
provide evidence that a4/01 may additionally have a role
in allergy, asthma, and autoimmune disorders (Walsh et
al., J. Immunol., 1991, 146:3419; Bochner et al., J.
Exp. Med., 1991 173:1553; Yednock et al., Nature, 1992,
356:63). Anti-a4 antibodies also block the migration of
leukocytes to the site of inflammation (Issedutz et al.,
J. Immunol., 1991, 147:4178).
The av/03 heterodimer is a member of the P3 integrin
subfamily and has been described on platelets,
endothelial cells, melanoma, smooth muscle cells, and
osteoclasts (Horton and Davies, J. Bone Min. Res. 1989,
4:803-808; Davies et al., J. Cell. Biol. 1989, 109:1817-
1826; Horton, Int. J. Exp. Pathol., 1990, 71:741-759).
Like GPIIb/IIIa, the vitronectin receptor binds a
variety of RGD-containing adhesive proteins such as
vitronectin, fibronectin, VWF, fibrinogen, osteopontin,
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bone sialo protein II and thrombosponden in a manner
mediated by the RGD sequence. A key event in bone
resorption is the adhesion of osteoclasts to the matrix
of bone. Studies with monoclonal antibodies have
implicated the av/P3 receptor in this process and suggest
that-a selective av/p3 antagonist would have utility in
blocking bone -resorption (Horton et al., J. Bone Miner.
Res., 1993, 8:239-247; Heif rich et al., J. Bone Miner.
Res., 1992, 7:335-343).
European Patent Application Publication Number
525629 (corresponds to Canadian Patent Application
Publication Number 2,074,685) discloses compounds ha`ving
the general formula:
x,,
xs, x2
A-B-C Xa 3 D-E-F
Copending, commonly assigned U.S. Patent
Application Serial Number 08/337,920 file6 11/10/94
discloses integrin inhibitors of the general formula
shown below:
R16 4 b O
Ri4 5
31 w-x-~
R'-U-V N..--O Y
PCT Patent Application WO 94/08577 published
4/28/94 discloses fibrinogen antagonists, including the
isoxazole-containing compound below:
-5-
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O C02H
H NHSO2Ph
HN
O NO
None of the above references teaches or suggests
the compounds of the present invention which are
described in detail below.
SLTNMARY OF THE INVENTION
The present invention provides novel nonpeptide
compounds which bind to integrin receptors thereby
altering cell-matrix and cell-cell adhesion processes.
The compounds of the present invention are useful for
the treatment of angiogenic disorders, inf lammation,
bone degradation, tumors, metastases, thrombosis, and
other cell aggregation-related conditions in a mammal.
One aspect of this invention provides novel
compounds of Formula I (described below) which are
useful as antagonists of the av/03 or vitronectin
receptor. The compounds of the present invention
inhibit the binding of vironectin to av/P3 and inhibit
cell adhesion. The present invention also includes
pharmaceutical compositions containing such compounds of
Formula I, and methods of using such compounds for the
inhibition of angiogenesis, and/or for the treatment of
angiogenic disorders.
The present invention also provides novel
compounds, pharmaceutical compositions and methods which
may be used in the treatment or prevention of diseases
which involve cell adhesion processes, including, but
not limited to, rheumatoid arthritis, asthma, allergies,
adult respiratory distress syndrome, graft versus host
disease, organ transplantation, septic shock, psoriasis, =
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eczema, contact dermatitis, osteoporosis,
osteoarthritis, atherosclerosis, metastasis, wound
healing, diabetic retinopathy, ocular vasculopathies,
thrombosis, inflammatory bowel disease and other
autoimmune diseases.
Also included in the present invention are
pharmaceutical kits comprising one or more containers
containing pharmaceutical dosage units comprising a
compound of Formula I, for the treatment of cell
adhesion related disorders, including, but not limited
to, angiogenic disorders.
DETAILED DESCRIPTION OF THE INVENTION
The present invention provides novel nonpeptide
compounds of Formula I (described below) which bind to
integrin receptors thereby altering cell-matrix and
cell-cell adhesion processes. The compounds of the
present invention are useful for the treatment of
angiogenic disorders, inflammation, bone degradation,
tumors, metastases, thrombosis, and other cell
aggregation-related conditions in a mammal.
One aspect of this invention provides novel
compounds of Formula I (described below) which are
useful as antagonists of the av/03 or vitronectin
receptor. The compounds of the present invention
inhibit the binding of vitronectin to av/03 and inhibit
cell adhesion. The present invention also includes
pharmaceutical compositions containing such compounds of
Formula I, and methods of using such compounds for the
inhibition of angiogenesis, and/or for the treatment of
angiogenic disorders.
[1] The present invention comprises compounds of the
Formula I:
-7-
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R1$
4 b
R14
g\ W X Y
R' U V O (I)
including stereoisomeric forms thereof, or mixtures of
stereoisomeric forms thereof, or pharmaceutically
acceptable salt or prodrug forms thereof wherein:
b, the bond between carbon atoms numbered 4 and 5, is a
carbon-carbon single or double bond;
Rl is selected from:
%-p- i ~ r N-M
' ~ A' ~ 11, L B ~ . B1'~ J=K
O
NH
/~ \4 F R2aN I
F~E p~E /
N N
= H
NHR12 NHR12
`N "\F
" p I or <
F~,E N~E
A and B are independently -CH2-, -0-, -N(R12)-, or
-C(=O)-;
Al and B1 are independently -CH2- or -N(Rlo)-; D is -N(R2a)-, -0-, -S-, -C(=0)-
or -SO2-;
-8-
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E-F is -C(R2)=C(R3) -, -N=C(R2) -, -C(R2)=N-, -N=N-, or
-C(R2)2C(R3)2-;
J, K, L and M are independently selected from -C(R2)- or
--N-, provided that at least one of J, K, L and M is
-C(R2)-;
R2 and R3 are independently selected from: H, C1-C4
alkoxy, NR11R12, =NR12, halogen, NO2 , CN, CF3 , C1-C6
alkyl, C3-C6 alkenyl, C3-C7 cycloalkyl, C4-C11
cycloalkylalkyl, C6-Cl0 aryl, C7-C11 arylalkyl,
C2 - C7 alkylcarbonyl, C6 - C10 carbonyl or C7- Cll
arylcarbonyl;
alternatively, R2 and R3, when substituents on
adjacent atoms, can be taken together with the
carbon atoms to which they are attached to form a
5-7 membered carbocyclic or 5-7 membered
heterocyclic aromatic or nonaromatic ring system,
said carbocyclic or heterocyclic ring being
optionally substituted with 0-2 R7;
R2a is absent or R12;
U is selected from:
- (CH2)n-,
- (CH2)nO(CH2)m-,
- (0H2)n,N(R12) (CH2)m-,
- (CH2)nC(=0) (CH2)m-,
- (CH2)nS(O)p(CH2)m-,
- (CH2)nNHNH(CH2)m-,
-N(R10)C(=0)-, or
C(=0)N(R10) -;
-N (Ri0) S (O) p- , or
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v is selected from:
- (CH2)n-,
-(C1-C6 alkylene)-Q-, substituted with 0-3 groups
'independently selected from R13,
-(C2-C7 alkenylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(C2-C7 alkynylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(phenyl)-Q-, said phenyl substituted with 0-2
groups independently selected from R13,
-(pyridyl)-Q-, said pyridyl substituted with 0-2
groups independently selected from R13, or
-(pyridazinyl)-Q-, said pyridazinyl substituted
with 0-2 groups independently selected from
R13;
Q is selected from:
- (CH2)n_ ,
- (CH2)n0(CH2)m-,
- (CH2)nN(R12) (CH2)m-,
- (CH2)nC(=O) (CH2)m-,
- (CH2) nS (O)p(CH2)m-,
- (CH2)nNHNH(CH2)m
-N(RlO) C(=0) -, or
-C(=0)N(Rl0) -;
W is selected from:
- (C(R4)2)QC(=O)N(Rlfl) -,
-C(=0) -N(RlO) - (C(R4) 2)q-;
X is selected from:
a single bond (i.e., X is absent),
- (C (R4) 2) q- LC (R4) (R8) l g-C (R4) (R9) - ;
:
alternatively, W is
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- (CHp)qC(=O)-N~ JZ
= and X is absent or -CH2-
Y is selected from:
-COR20, -SO3H, -PO3H, -CONHNHSO2CF3, -CONHSO2R18a,
-CONHSO2NHR18b, -NHCOCF3, -NHCONHSO2R18a,
-NHSO2R18a' -OP03H2, -OSO3H, -P03H2, -SO3H,
-SO2NHCOR18a, -gO2NHCO2R18a, or
N O
I~N ~ AN
N ~ CF3
- Nj _ _
I
H H or HO O
Z is selected from -CH(R9) -, or -N(R16) -;
R4 is selected from H, C1-Clo alkyl, C1-Clo
alkylcarbonyl, aryl, arylalkyl, cycloalkyl, or
cycloalkylalkyl;
alternatively, two R4 groups on adjacent carbon
atoms may join to form a bond, thereby to form a
carbon-carbon double or triple bond between the
adjacent carbon atoms;
RS is selected from H, C1-C6 alkyl, C2-C6 alkenyl, C2-
C6 alkynyl, C3-C7 cycloalkyl, C7-C14 bicycloalkyl,
hydroxy, C1-C6 alkoxy, C1-C6 alkylthio, C1-C6
alkylsulfinyl, C1-C6 alkylsulfonyl, nitro, C1-C6
alkylcarbonyl, C6-Clo aryl, -N(R11)R12, halo, CF3,
CN, C1-C6 alkoxycarbonyl, carboxy, piperidinyl,
morpholinyl or pyridinyl;
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R6 is selected from:
H, Cl-C10 alkyl, hydroxy, Cl-C10 alkoxy, nitro, Cl-
C10 alkylcarbonyl, -N(Rll)R12, cyano, halo,
CF3, CHO, CO2Rl8b, C(=O) RlBb, CONR17R18b, 5 OC (=0) R10, OC (=0) OR21, OR10,
OC (=O) NR10R11 ,
OCH2C02R10, C02CH2CO2R10, NO2, NRlOC(=O)R10, NR10C (=O) OR21, NR10C (=0 )
NR10g.11 , Ng10802NR1OR11,
NRlOSO2R21, S( O) pRll , S02NR10R11 , SiMe3, C2 to
C6 alkenyl, C3 to C11 cycloalkyl, C4 to C11
cycloalkylmethyl,
C6 to C10 aryl optionally substituted with 1-3
groups selected from halogen, Cl-C6 alkoxy,
Cl-C6 alkyl, CF3, S(O)mMe, or -NMe2;
C7 to C11 arylalkyl, said aryl being optionally
substituted with 1-3 groups selected from
halogen, Cl-C6 alkoxy, C1-C6 alkyl, CF3,
S(O)pMe, or -NMe2,
methylenedioxy when R6 is a substituent on aryl, or
a 5-10 membered heterocyclic ring containing 1-3 N,
0, or S heteroatoms, wherein said heterocyclic
ring may be saturated, partially saturated, or
fully unsaturated, said heterocyclic ring
being substituted with 0-2 R7;
R7 is selected from:
H, Cl-C10 alkyl, hydroxy, Cl-C10 alkoxy, nitro,
Cl-C10 alkylcarbonyl, -N(R11)R12, cyano, halo,
CF3, CHO, CO2R10, C(=O) R10 , CONR10R11,
OC (=O) R10, OC (=0) OR21, OR10, OC (=0) NR10R11 ,
OCH2CO2R10, CO2CH2CO2R10, NO2, NRlOC(=O)R10,
NRlOC (=O) OR21, NRlOC (=0) NR10R11 , NR10S02NR10R11,
NR10S02R21, S(O)pRll, SO2NR10R11, SiMe3, C2 to
C6 alkenyl, C3 to C11 cycloalkyl, C4 to C11
cycloalkylmethyl, C6 to C10 aryl, or C7 to C11
arylalkyl;
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RS is selected from:
H. R6,
C1-C10 alkyl, substituted with 0-3 R6,
C2-Cl0 alkenyl, substituted with 0-3 R6,
C2-C10 alkynyl, substituted with 0-3 R6,
C3-CS cycloalkyl, substituted with 0-3 R6,
Cg-C6 cycloalkenyl, substituted with 0-3 R6,
aryl, substituted with 0-3 R6, or
5-10 membered heterocyclic ring containing 1-3 N,
0, or S heteroatoms, wherein said heterocyclic
ring may be saturated, partially saturated, or
fully unsaturated, said heterocyclic ring
being substituted with 0-2 R7;
R9 is selected from H, hydroxy, C1-Clp alkoxy, nitro,
N(R10) R11, -N(R16) R17 , OR22, Cl-Clp alkyl substituted
with 0-3 R7, aryl substituted with 0-3 R7,
heteroaryl substituted with 0-3 R7, Cl-Clo
alkylcarbonyl; aryl(Co-C6 alkyl)carbonyl, C1-Clo
alkenyl, C1-Clp alkynyl ,C3-Clp cycloalkyl, C3-Clo
cycloalkylalkyl, aryl(Ci-C6 alkyl) -, heteroaryl(C1-
Cs alkyl)-, CO2Ri8a, C(=O)Rl8a, CONR18aR20, S0ZR18a,
or SO2NRl8ap20 , provided that any of the above
alkyl, cycloalkyl, aryl or heteroaryl groups may
be unsubstituted or substituted independently with
0-2 R7;
R10 is selected from H, C1-C8 alkyl, C3-C6 alkenyl,
C3-C11 cycloalkyl, C4-C11 cycloalkylmethyl, C6-C1o
aryl, C7-C11 arylalkyl, or Cl-Clo alkyl substituted
with 0-2 R4;
R11 is selected from hydrogen, hydroxy, C1 to C8 alkyl,
C3-C6 alkenyl, C3 to C11 cycloalkyl, C4 to C11
cycloalkylmethyl, C1-C6 alkoxy, bentyloxy, C6 to C10
aryl, heteroaryl, heteroarylalkyl, C7 to C11
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arylalkyl, adamantylmethyl, or Cl-C10 alkyl
substituted with 0-2 R4;
alternatively, Rio and R" when both are substituents on =
the same nitrogen atom (as in -NRlORll) can be taken
together with the nitrogen atom to which they are
attached,to form a heterocycle selected from:
3-azabicyclononyl, 1,2,3,4-tetrahydro-l-quinolinyl,
1,2,3,4-tetrahydro-2-isoquinolinyl, 1-piperidinyl,
1-morpholinyl, 1-pyrrolidinyl, thiamorpholinyl,
thiazolidinyl or 1-piperazinyl; said heterocycle
being optionally substituted with 0-3 groups
selected from: C1-C6 alkyl, C6-Clo aryl, heteroaryl,
C7-C11 arylalkyl, C1-C6 alkylcarbonyl, C3-C7
cycloalkylcarbonyl, C1-C6 alkoxycarbonyl, C7-C11
arylalkoxycarbonyl, C1-C6 alkylsulfonyl or C6-Clo
aryl sul f onyl ;
R12 is selected from:
H, Cl-Clo alkyl, triphenylmethyl,
methoxyphenyldiphenylmethyl,
trimethylsilylethoxymethyoxy (SEM), C1-Clo
alkoxycarbonyl, C1-C10 alkylcarbonyl, C1-Clo
alkylsulfonyl, aryl (C1-Clo alkyl) sulfonyl,
arylsulfonyl, aryl (C2-Clo alkenyl) sulfonyl,
heteroarylsulfonyl, aryl, C2-C6 alkenyl, C3-C11
cycloalkyl, C4-C11 cycloalkylalkyl, C7-C11
arylalkyl, C7-C11 arylcarbonyl, C4-C11
cycloalkoxycarbonyl, C7-C11 bicycloalkoxycarbonyl,
C7-C11 aryloxycarbonyl, heteroarylcarbonyl,
heteroarylalkylcarbonyl, or
aryl(C1-Cl0 alkoxy)carbonyl; wherein said aryl
groups are optionally substituted with 0-3
substituents selected from the group consisting of:
C1-C4 alkyl, Cl-C4 alkoxy, halo, CF3, and NO2;
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R13 is selected from: H, hydroxy, C1-C10 alkoxy, nitro,
N(R10)R11, -N(R16)R17, C1-Clo alkyl substituted with
0-3 R7, aryl substituted with 0-3 R7, heteroaryl
substituted with 0-3 R7, or C1-C10 alkylcarbonyl;
R14 is selected from H, C1-C14 alkyl, C2-C10 alkenyl,
C2-C10 alkynyl, C1-Clo alkoxy, aryl, heteroaryl or
C1-C10 alkoxycarbonyl, CO2R10 or -C(=0)N(R10)R11;
R15 is selected from:
H, R6, -CO2R10, -C (=0) N(R10) Rll;
C1-C10 alkoxycarbonyl substituted with 0-2 R6;
C1-C10 alkyl, substituted with 0-3 R6;
C2-C10 alkenyl, substituted with 0-3 R6;
C1-C10 alkoxy, substituted with 0-3 R6;
aryl, substituted with 0-3 R6; or
5-10 membered heterocyclic ring containing 1-3 N,
0, or S heteroatoms, wherein said heterocyclic
ring may be saturated, partially saturated, or
fully unsaturated, said heterocyclic ring
being substituted with 0-2 R7;
R16 is selected from:
-C(=O) -0-R18a,
-C(=O) -R18b,
-C(=0)N(R18b)2,
-C(=0)NHSO2R18a,
-C (=0) NHC (=0) R18b,
- C ( =O ) NHC ( =0 ) OR18 a ,
-C (=0) NHSO2NHR18b,
-C(=S) -NH-R18b,
-NH-C (=0) -O-R18a,
-NH-C (=0) -R18b,
-NH-C (=0) -NH-R18b,
-S02-0-R18a,
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-S02-R18a,
-S02-N(R18b)2,
-S02-NHC (=0) OR18b;
R3-7 is selected from: H, C1-Clo alkyl, C2-C6 alkenyl, C3-
C11 cycloalkyl, C4-C15 cycloalkylalkyl, aryl,
aryl(C1-Cip alkyl) - ;
R18a is selected from:
C1-C8 alkyl substituted with 0-2 R19,
C2-C8 alkenyl substituted with 0-2 R19,
C2-C8 alkynyl substituted with 0-2 R19,
C3-C8 cycloalkyl substituted with 0-2 R19,
aryl substituted with 0-4 R19,
aryl(Cl-C6 alkyl)- substituted with 0-4 R19,
a 5-10 membered heterocyclic ring system having 1-3
heteroatoms selected independently from 0, S,
and N, said heterocyclic ring being
substituted with 0-4 R19,
C1-C6 alkyl substituted with a 5-10 membered
heterocyclic ring system having 1-3
heteroatoms selected independently from 0, S,
and N, said heterocyclic ring being
substituted with 0-4 R19;
R18b is selected from R18a or H;
R19 is selected f rom H, halogen, CF3, CO2H, CN, NO2,
NR11R12, C1-C8 alkyl, C2-C6 alkenyl, C2-C6 alkynyl,
C3-C11 cycloalkyl, C4-C11 cycloalkylalkyl,
aryl(Cl-C6 alkyl)-, C1-C6 alkoxy, OCF3, or C1-C4
alkoxycarbonyl, aryl, -O-aryl, -S02-aryl,
heteroaryl, or -S02-heteroaryl, wherein said aryl
and heteroaryl groups may be substituted with 0-4
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groups selected from hydrogen, halogen, CF3, C1-C3
alkyl, or C1-C3 alkoxy;
R20 is selected from hydroxy, C1 to CIO alkyloxy, C3 to
C1,i cycloalkyloxy, C6 to C10 aryloxy, C7 to C11
arylalkyloxy, C3 to CIO alkylcarbonyloxyalkyloxy, C3
to CIO alkoxycarbonyloxyalkyloxy, C2 to Clo
s,lkoxycarbonylalkyloxy, CS to CIO
cycloalkylcarbonyloxyalkyloxy, CS to CIO
cycloalkoxycarbonyloxyalkyloxy, C5 to CIO
cycloalkoxycarbonylalkyloxy, C7 to C11
aryloxycarbonylalkyloxy, C8 to C12
aryloxycarbonyloxyalkyloxy, C8 to C12
arylcarbonyloxyalkyloxy, C5 to C1o
alkoxyalkylcarbonyloxyalkyloxy, C5 to CIO (5-alkyl-
1,3-dioxa-cyclopenten-2-one-yl)methyloxy, C10 to C14
(5-aryl-l,3-dioxa-cyclopenten-2-one-yl)methyloxy,
(R11) (R12)N- (Cl - Clo alkoxy) -;
R21 is selected from: C1-C8 alkyl, C2-C6 alkenyl, C3-C11
cycloalkyl, C4-C11 cycloalkylmethyl, C6-C10 aryl,
C7-C11 arylalkyl, or C1-C10 alkyl substituted with
0-2 R5;
R22 is selected from:
-C (=O) -R18b,
-C(=0)N(R18b)2,
-C(=0)NHSO2R18a,
-C (=0) NHC (=0) R18b,
-C (=0) NHC (=O) OR18a,
- C (=O ) NHSO2NHR18b,
- C (=S) - NH - R18b,
-S02-R18a,
-S02-N(R18b)2,
S02-NHC (=0) OR18b
-17-
CA 02221980 1997-11-24
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m is 0-2;
n is 0-4;
p is 0-2;
q is 0-4;
r is 0-2;
s is 0-1;
with the following provisos:
(1) when b is a double bond, only one of R14 or R15
is present and Q and U are not -(CH2)-; and
(2) n, m and q are chosen such that the number of
atoms connecting R1 and Y is in the range of
8-14; and
(3) when V is -(phenyl)-Q-, then either: U is not a
direct bond (i.e., U is not -(CH2)n- where n
0) or Q is not a direct bond (i.e., Q is not
- ( CH2 ) n- where n = 0 ) .
[2] Preferred compounds of the present invention are
compounds of Formula 2:
R15
4 b
R14
5
3 W X Y
R1 U VN--'O
(I)
including stereoisomeric forms thereof, or mixtures of
stereoisomeric forms thereof, or pharmaceutically
acceptable salt or prodrug forms thereof wherein:
b, the bond between carbon atoms numbered 4 and 5, is a
carbon-carbon single or double bond;
-18-
CA 02221980 1997-11-24
W O 96/37492 PCT/US96/07646
R1 is selected from:
N N--~ r N-M
A' ~ L
= A r B-< J=K
NH 0 NHR12
.N~F
I ' N ' N D
~
,
D-E I 1
NHR12
iXTJor N ~ N H
A is selected from -CH2-, or -N(Rl2) -;
Al and B are independently -CH2- or -N(Rlo)-;
D is -N(R12)-, or -S-;
E-F is -C(R2)=C(R3) -, or -C(R2)2C(R3)2-;
J is either -C(R2)- or -N-, and K, L and M are
independently selected from -C(R2) - or -C(R3) -;
R2 and R3 are independently selected from: H, Cl-C4
alkoxy, NR11R12, =NR12, halogen, NO2, CN, CF3, Cl-C6
alkyl, C3-C6 alkenyl, C3-C7 cycloalkyl, C4-C11
cycloalkylalkyl, C6-C10 aryl substituted with 0-4
R7, C7-C11 arylalkyl, C2-C7 alkylcarbonyl, CI-C4
alkoxycarbonyl, or C7-C11 arylcarbonyl;
alternatively, R2 and R3 when substituents on
adjacent atoms, can be taken together when
-19-
CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
substituents on adjacent atoms, with the carbon
atoms to which they are attached, to form a 5-7
membered carbocyclic or 5-7 membered heterocyclic
with the carbon atoms to which they are attached,
aromatic or nonaromatic ring system, said
carbocyclic or heterocyclic ring being optionally substituted with 0-2 groups
selected from Cl-C4
alkyl, C1-C4 alkoxy, halo, cyano, amino, CF3 or NO2;
R2a is absent or R12;
U is selected from:
- (CH2)n-,
- (CH2)nO(CH2)m
- (CH2)nN(R12) (CH2)m-,
- ( CH2 ) nC ( =O ) (CH2 ) m - ,
- (CH2)nS (O)p(CH2)m-,
- (CH2)nNHNH(CH2)m-,
-N (R10) C (=0) - , or
-C(=0)N(R10) -;
-N (Ri0) S (O) p- , or
V is selected from:
- (CH2) n- ,
-(Cl-C6 alkylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(C2-C7 alkenylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(C2-C7 alkynylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(phenyl)-Q-, said phenyl substituted with 0-2
groups independently selected from R13,
-(pyridyl)-Q-, said pyridyl substituted with 0-2
groups independently selected from R13, or
-20-
CA 02221980 1997-11-24
WO 96137492 PCT/US96107646
-(pyridazinyl)-Q-, said pyridazinyl substituted
with 0-2 groups independently selected from
R13;
Q is selected from:
- (C112) n - ,
( CH2 ) nO ( CH2 ) m
- ~CH2)nN(R12) (CH2)m-,
- ( CH2 ) nC ( =O ) ( CH2 ) m - ,
- (CH2)nS(O)p(CH2)m-,
- (CH2)nNHNH(CH2)m-,
-N (R10) C (=O) - , or
-C(=0)N(R10) -;
w is selected from:
- (C(R4)2)qC(=O)N(R10) - or,
-C (=0) -N(R10) - (C (R4) 2) q-;
X is selected from:
a single bond (i.e., X is absent) or,
- (C (R4) 2) q- [C (R4) (R8) I 8-C (R4) (R9) - :
alternatively, W is
-(CH2)qC(-O)-N Z
` J
I
and X is either absent or -CH2-
Y is selected from:
-COR20, -SO3H, -PO3H, -CONHNHSO2CF3,
-CONHSO2Rl8a, -CONHSO2NHR18b, -NHCOCF3,
-NHCONHS02R18a, -NHSO2R18a, -OP03H2, -OSO3H,
-P03H2, -SO3H, -SO2NHCOR18a, -S02NHCO2R18a, Oz.
-21-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
~N ~-~1 O
CF3
)L2N
H H I-10 O
or
Z is selected from -CH(R9) -, or -N(R16) -;
R4 is selected from H, C1YClo alkyl, C1-Ci,o
alkylcarbonyl, aryl, arylalkyl, cycloalkyl, or
cycloalkylalkyl;
alternatively, two R4 groups on adjacent carbon
atoms may join to form a bond, thereby to form a
carbon-carbon double or triple bond between the
adjacent carbon atoms;
R6 is selected from:
H, C1-Clo alkyl, hydroxy, C1-C,i0 alkoxy, nitro, C1-
Clo alkylcarbonyl, -N(Ril)R12, cyano, halo,
CF3, CHO, CO2R18b, C(=O) RIBb, CONR17R18b,
OC(=0)R10, OR10, OC(=O)NR10R11, NR10C(=O)R10,
NR10C (=O) OR21, NRlOC (=0) NR10R11 , NR10502NR10R11,
NR10S02R21, S (O) pRll, SO2NR10RI1,
C6 to Clo aryl optionally substituted with 0-3
groups selected from halogen, C1-C6 alkoxy,
C1-C6 alkyl, CF3, S(O)mMe, or -NMe2;
C7 to C11 arylalkyl, said aryl being optionally
substituted with 1-3 groups selected from
halogen, C,i - C6 alkoxy, C1, - C6 alkyl, CF3,
S(O)pMe, or -NMe2,
a 5-10 membered heterocyclic ring containing 1-3 N,
0, or S heteroatoms, wherein said heterocyclic
ring may be saturated, partially saturated, or
fully unsaturated, said heterocyclic ring
being substituted with 0-2 R7;
-22-
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CA 02221980 1997-11-24
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R7 is selected from selected from H, C,i-C4 alkyl,
hydroxy, Cl - C4 alkoxy, C6 - C,io aryl, C7- Cli
arylalkyl, (Cl-C4 alkyl)carbonyl, CO2R18a, SO:2Rli,
SO2NR10R11-, OR10, or N(Rl-1) R12;
R8 "is selected from:
H, CO-Rlsa, C(=0) Rl8a, or CONRJ7R18a
CI-C10 alkyl, substituted with 0-1 R6,
C2-Clo alkenyl, substituted with 0-1 R6,
C2-C,to alkynyl, substituted with 0-1 R6,
C3-C8 cycloalkyl, substituted with 0-1 R6,
C5-C6 cycloalkenyl, substituted with 0-1 R6,
aryl, substituted with 0-3 R6, or
5-10 membered heterocyclic ring containing 1-3 N,
0, or S heteroatoms, wherein said heterocyclic
ring may be saturated, partially saturated, or
fully unsaturated, said heterocyclic ring
being substituted with 0-2 R7;
R9 is selected from H, hydroxy, Cl-Clo alkoxy, nitro,
N(Rl0) R11, -N(R16)R17, OR22, Cl,-Clo alkyl substituted
with 0-3 R7, aryl substituted with 0-3 R7,
heteroaryl substituted with 0-3 R7, Cl-Clo
alkylcarbonyl; aryl(Co-C6 alkyl)carbonyl, C1-Clo
alkenyl, C3,-C,io alkynyl ,C3-Clo cycloalkyl, C3-C10
cycloalkylalkyl, aryl(Cl-C6 alkyl) - , heteroaryl(C1-
C6 alkyl)-, C02R18a. C(=O)RlBa, CONR18aR20, S02R18a,
or SO2NR18aR20, provided that any of the above
alkyl, cycloalkyl, aryl or heteroaryl groups may
be unsubstituted or substituted independently with
0-2 R7 ;
Rlo is selected from H, Cl-C8 alkyl, C3-C6 alkenyl,
C3-C11 cycloalkyl, C4-C11 cycloalkylmethyl, C6-Clo
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
aryl, C7-C11 arylalkyl, or C1-C10 alkyl substituted
with 0-2 R4;
R11 is selected from hydrogen, hydroxy, Ci, to Cs alkyl,
C3-C6 alkenyl, C3 to C11 cycloalkyl, C4 to C11
cycloalkylmethyl, C1-C6 alkoxy, benzyloxy, C6 to Clo
aryl, heteroaryl, heteroarylalkyl, C7 to C11
arylalkyl, adamantylmethyl, or Cl-Clo alkyl
substituted with 0-2 R4;
alternatively, RlO and R11 when both are substituents on
the same nitrogen atom (as in -NRlORll) can be taken
together with the nitrogen atom to which they are
attached to form a heterocycle selected from:
3-azabicyclononyl, 1,2,3,4-tetrahydro-l-quinolinyl,
1,2,3,4-tetrahydro-2-isoquinolinyl, 1-piperidinyl,
1-morpholinyl, 1-pyrrolidinyl, thiamorpholinyl,
thiazolidinyl or 1-piperazinyl; said heterocycle
being optionally substituted with 0-3 groups
selected from: C1-C6 alkyl, C6-Clp aryl, heteroaryl,
C7-C11 arylalkyl, Cl-C6 alkylcarbonyl, C3-C7
cycloalkylcarbonyl, C1-C6 alkoxycarbonyl, C7-C11
arylalkoxycarbonyl, C1-C6 alkylsulfonyl or C6-Clo
aryl su1 f ony1;
R12 is selected from:
H, C1-C6 alkyl, triphenylmethyl,
methoxyphenyldiphenylmethyl,
trimethylsilylethoxymethyoxy (SEM), (Cl-C6
alkyl)carbonyl, (C1-C6 alkoxy)carbonyl; (C1-C6
alkyl) amino carbonyl, C3-C6 alkenyl, C3-C7
cycloalkyl, C4-C11 cycloalkylalkyl, aryl,
heteroaryl(Cz-C6 alkyl)carbonyl,
heteroarylcarbonyl, aryl C1-C6 alkyl, (C1-C6 35 alkyl)carbonyl, or
arylcarbonyl, CI-C6
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
alkylsulfonyl, arylsulfonyl, aryl(C1-C6
alkyl)sulfonyl, heteroarylsulfonyl,
heteroaryl(C1-C6 alkyl)sulfonyl, aryloxycarbonyl,
or aryl(C1-C6 alkoxy)carbonyl, wherein said aryl
groups are substituted with 0-2 substituents
selected from the group consisting of C1-C4 alkyl,
C1-C4 alkoxy, halo, CF3, and nitro;
R13 is selected from: H, hydroxy, C1-C10 alkoxy, nitro,
N( R10 ) R11 , - N( R16 ) Ri7 , Ci - C10 alkyl substituted with
0-3 R7, aryl substituted with 0-3 R7, heteroaryl
substituted with 0-3 R7, or C1-Clo alkylcarbonyl;
R14 is selected from H. C1-C10 alkyl, C2-Cip alkenyl,
C2-Clo alkynyl, C1-Clo alkoxy, aryl, heteroaryl or
Ci-Cip alkoxycarbonyl, CO2R10 or -C (=O) N(Rlo) Rll;
R15 is selected from: H, CO2R18a, C(=O) R18a, CONR18aR17 r
-S02R18a, -S02NR18aR17, C1-C6 alkyl substituted with
0-1 R9, C3-C6 alkenyl substituted with 0-1 R9, C3-C7
cycloalkyl substituted with 0-1 R9, C4-C11
cycloalkylalkyl substituted with 0-1 R9, aryl
substituted with 0-1 R9 or 0-2 R7, or aryl(C1-C6
alkyl)- substituted with 0-1 R9 or 0-2 R7;
R16 is selected from:
-C (=O) -O-R18a,
-C (=O) -R18b,
-C(=O)N(R18b)2~
-C(=O)NHSO2R18a,
- C ( =O ) NHC ( =O ) R18b,
-C (=0) NHC (=0) OR18a,
-C(=O)NHSO2NHR18b,
-S02-R18a,
-SO2-N(R18b)2 or,
S02 - NHC (=0 ) OR18b;
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CA 02221980 1997-11-24
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R17 is selected from: H, Cl-C6 alkyl, C3-C7 cycloalkyl,
C4-C11 cycloalkylalkyl, aryl, aryl(Cl-C5 alkyl)-, or
heteroaryl(Cl-C6 alkyl);
R18a is selected from: C1-CO alkyl, C3-C11 cycloalkyl,
aryl ( Cl - C6 alkyl)-, ( Cz - C6 alkyl) aryl ,
heteroaryl(C1-C6 alkyl)-, (C1-C6 alkyl)heteroaryl,
biaryl(Cl-C6 alkyl), heteroaryl, or aryl, wherein
said aryl or heteroaryl groups are optionally
substituted with 0-4 R19;
R18b is selected from R18a or H;
R19 is selected from H, halogen, CF3, CO2H, CN, NO2,
NR11R12, Cl-C8 alkyl, C2-C6 alkenyl, C2-C6 alkynyl,
C3-C11 cycloalkyl, C4-C11 cycloalkylalkyl,
aryl(C1-C6 alkyl)-, C1-C6 alkoxy, OCF3, or Cl-C4
alkoxycarbonyl, aryl, -0-aryl, -SO2-aryl,
heteroaryl, or -S02-heteroaryl, wherein said aryl
and heteroaryl groups may be substituted with 0-4
groups selected from hydrogen, halogen, CF3, C1-C3
alkyl, or Cl-C3 alkoxy;
R20 is selected from hydroxy, C1 to C10 alkyloxy, C3 to
Cli, cycloalkyloxy, C6 to Clo aryloxy, C7 to C11
aralkyloxy, C3 to Clp alkylcarbonyloxyalkyloxy, C3
to Clp alkoxycarbonyloxyalkyloxy, C2 to C10
alkoxycarbonylalkyloxy, CS to C1Q
cycloalkylcarbonyloxyalkyloxy, C5 to C1o
cycloalkoxycarbonyloxyalkyloxy, C5 to Clo
cycloalkoxycarbonylalkyloxy, C7 to C11
aryloxycarbonylalkyloxy, CS to C12
aryloxycarbonyloxyalkyloxy, C8 to C12
arylcarbonyloxyalkyloxy, C5 to C10
alkoxyalkylcarbonyloxyalkyloxy, C5 to Clo (5-alkyl-
-26-
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CA 02221980 1997-11-24
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1,3-dioxa-cyclopenten-2-one-yl)methyloxy, Clp to C14
(5-aryl-1,3-dioxa-cyclopenten-2-one-yl)methyloxy,
or (Rll) (R12)N- (C1-Clp alkoxy) -;
R21 is selected from: C1-CS alkyl, C2-C6 alkenyl, C3-C11
cycloalkyl, C4-Cli1 cycloalkylmethyl, C6-C10 aryl,
C7-C11 arylalkyl, or Cl-Clp alkyl substituted with
0-2 R7;
R22 is selected from:
-C(=0) -R18b,
-C(=0)N(R18b)
-C (=0) NHSO`RiBa,
- C (=O) NHC (=O) R18b,
-C (=0) NHC (=O) ORiBa or,
-C (=0) NHSOrI'7HR18b,
m is 0-2;
n is 0-4;
p is 0-2;
q is 0-4;
r is 0-2;
s is 0-1;
with the following provisos:
(1) when b is a double bond, only one of R14 or R15
is present and Q and U are not -(CH2)-; and
(2) n, m and q are chosen such that the number of
atoms connecting Rl and Y is in the range of
8-14; and
(3) when v is -(phenyl)-Q-, then either: U is not a
direct bond (i.e., U is not -(CH2)n- where n
0) or Q is not a direct bond (i.e., Q is not
- (CH2 ) n- where n = 0).
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
L31 Further preferred compounds of the above invention
are compounds of Formula II:
4 b
R14 11~
-. 5
3 `' W X Y '
R1 U O
{II)
including enantiomeric or diastereomeric forms thereof,
or mixtures of enantiomeric or diastereomeric forms
thereof, or pharmaceutically acceptable salt or prodrug
forms thereof wherein:
b, the bond between carbon atoms numbered 4 and 5, is a
carbon-carbon single or double bond;
{
-28-
CA 02221980 1997-11-24
WO 96137492 PCT/1JS96/07646
Rl is selected from:
R2 CN R2
C
N N
N~--- N :~c
R3 H H R3 H
N CN N _ N N N
~ N~
N ,
R12NH~N R12NHN
[ N
N ! Z~/ HN ~
,
R12N R12NH
N N
N CN O1,orCTO
N H H
R2 and R3 are independently selected from: H, Cl-C4
alkoxy, NRlIR12 , halogen, N02, CN, CF3, Cl - C6 alkyl,
C3-C6 alkenyl, C3-C7 cycloalkyl, C4-C11
cycloalkylalkyl, C6-Clp aryl substituted with 0-2
R7, C7-C11 arylalkyl, C2-C7 alkylcarbonyl, or C7-C11
arylcarbonyl;
alternatively, R2 and R3 can be taken together with
the carbon atoms to which they are attached to form
a 5-7 membered carbocyclic or 5-7 membered
-29-
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
heterocyclic aromatic or nonaromatic ring system,
said carbocyclic or heterocyclic ring being
optionally substituted with 0-2 R7;
U is selected from:
-- (CH`)n-,
-N(R12) (CH2)m-,
-i`T(R10) C (=O) - , or -C(=O)N(R10) -;
-N(R1O)S(O)p-, or
V is selected from:
- (CH2) n- ,
-(C1-C6 alkylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(C2-C7 alkenylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(C2-C7 alkynylene)-Q-, substituted with 0-3 groups
independently selected from R13,
-(phenyl)-Q-, said phenyl substituted with 0-2
groups independently selected from R13,
-(pyridyl)-Q-, said pyridyl substituted with 0-2
groups independentlyselected from R13, or
-(pyridazinyl)-Q-, said pyridazinyl substituted
with 0-2 groups independently selected from
R13;
Q is selected from:
- (CH2) n- ,
- (CH2)nO(CH2)m-,
- (CH2)nN(R12) (CH2)m-,
-N(R10) C(=O) -, or
-C(=O)N(R10) -;
W is selected from:
- (CH2)qC(=0)N(R10) -, or
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
-C(=0) -N(R10) - (CH2)~-;
x is - (0H2)Q-CH(R8) -CH(R9) -;
Y is -COR20;
R6 is selected'from:
H, Cl-C4 alkyl, hydroxy, Cl-C4 alkoxy, nitro, Cl-C6
alkylcarbonyl, -N (Rl1) R12, cyano, halo, CF3,
-S (O) pRlO, CO2Ri8a, CONR17R18a, -COR18a, ORlO,
C6 to Clp aryl optionally substituted with 0-3
groups selected from halogen, Cl-C6 alkoxy,
Cl-C6 alkyl, CF3, S(O)mMe, or -NMe2;
a heterocyclic ring system selected from pyridinyl,
furanyl, thiazolyl, thienyl, pyrrolyl,
pyrazolyl, triazolyl, imidazolyl,
benzofuranyl, indolyl, indolinyl, quinolinyl,
isoquinolinyl, benzimidazolyl, piperidinyl,
tetrahydrofuranyl, pyranyl, 3H-indolyl,
carbazolyl, pyrrolidinyl, piperidinyl,
isoxazolinyl, isoxazolyl, or morpholinyl;
R7 is selected from:
H, Cl-C4 alkyl, hydroxy, Cl-C4 alkoxy, nitro, Cl-C4
alkylcarbonyl, -N (Rll) Rl2 , CO2R1ea, S02R11,
SO2NRlORl1 or OR10;
R8 is selected from:
H, CONR17R18a, -C02R18a, -CORlBa
Cl-Clp alkyl, substituted with 0-1 R6,
C2-Clo alkenyl, substituted with 0-1 R6,
C2-Clp alkynyl, substituted with 0-1 R6,
C3-Cg cycloalkyl, substituted with 0-1 R6,
aryl, substituted with 0-1 R6 or,
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
a heterocyclic ring system selected from pyridinyl,
furanyl, thiazolyl, thienyl, pyrrolyl,
pyrazolyl, triazolyl, imidazolyl,
benzofuranyl, indolyl, indolinyl, quinolinyl,
isoquinolinyl, benzimidazolyl, piperidinyl,
tetrahydrofuranyl, pyranyl, 3H-indolyl,
carbazolyl, pyrrolidinyl, piperidinyl,
isoxazolinyl, isoxazolyl or morpholinyl, said
heterocycle optionally substituted with 0-2
R7;
R9 is selected from: H or -N(R16) R17;
Rlo is selected from H or C1-Clo alkyl, or C-7-Clo
arylalkyl;
R11 is selected from hydrogen, hydroxy, Cl to C$ alkyl,
C3-C6 alkenyl, C3 to C11 cycloalkyl, C4 to C11
cycloalkylmethyl, Cl-C6 alkoxy, benzyloxy, C6 to Clo
aryl, heteroaryl, heteroarylalkyl, C7 to C11
arylalkyl, adamantylmethyl, or Cl-Clo alkyl
substituted with 0-2 R4 ;
alternatively, Rlo and R" when both are substituents on
the same nitrogen atom (as in -NRloR11) can be taken
together with the nitrogen atom to which they are
attached to form a heterocycle selected from:
3-azabicyclononyl, 1,2,3,4-tetrahydro-l-quinolinyl,
1,2,3,4-tetrahydro-2-isoquinolinyl, 1-piperidinyl,
1-morpholinyl, 1-pyrrolidinyl, thiamorpholinyl,
thiazolidinyl or 1-piperazinyl; said heterocycle
being optionally substituted with 1-3 groups
selected from: C1-C6 alkyl, C6-Clo aryl, heteroaryl,
C7-C11 arylalkyl, Cl-C6 alkylcarbonyl, C3-C7
cycloalkylcarbonyl, Cl-C6 alkoxycarbonyl, C7-C11
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CA 02221980 1997-11-24
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arylalkoxycarbonyl, C1-C6 alkylsulfonyl or C6-Clo
aryl sulf onyl ;
R12 is selected from:
H, C1-C6 alkyl, triphenylmethyl,
methoxyphenyldiphenylmethyl,
trimethylsilylethoxymethyoxy (SEM), C1-C4
alkoxycarbonyl, Cl-C6 aikylcarbonyl, C1-C6
alkylsulfonyl, aryl(C1-C4 alkyl)sulfonyl,
arylsulfonyl, aryl, heteroarylcarbonyl, or
heteroarylalkylcarbonyl, wherein said aryl groups
are substituted with 0-3 substituents selected from
the group consisting of: C1-C4 alkyl, C1-C4 alkoxy,
halo, CF3, and NO2;
R13 is selected from: H, hydroxy, C1-Cla alkoxy,
N(RiO)Rll, -N(R16)Rl7, C1-Clp alkyl substituted with
0-3 R7, aryl substituted with 0-3 R7, heteroaryl
substituted with 0-3 R7, or C1-Clq alkylcarbonyl;
R14 is selected from H, C1-Clo alkyl, C2-Clo alkenyl,
C2-Clp alkynyl, C1-C10 alkoxy, aryl, heteroaryl or
C1-Clp alkoxycarbonyl, C02R10 or -C(=O)N(R1o)R11;
R16 is selected from:
-C (=O) -O-R18a,
-C (=O) -R18b,
-SO2-R18a or,
-SO2-N(18b)2;
R17 is selected from H or C1-C4 alkyl;
R18a is selected from: C1-C8 alkyl, C3-C11 cycloalkyl,
aryl (C1-CS alkyl) -, (C1-C6 alkyl) aryl,
heteroaryl ( C1- C6 alkyl )-, ( C1- C6 alkyl ) heteroaryl,
biaryl(C1-C6 alkyl), heteroaryl, or aryl, wherein
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CA 02221980 1997-11-24
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said aryl or heteroaryl groups are optionally
substituted with 0-2 R19;
R18b is selected from R18a or H;
R19 is selected from H, halogen, CF3, CO2H, CN, N02,
NR11R12. C1-Cg alkyl, C2-C6 alkenyl, C2-C6 alkynyl,
C3-C11, cycloalkyl, C4-C11 cycloalkylalkyl,
aryl(Cl-C6 alkyl)-, Cl-C6 alkoxy, OCF3, or Cl-C4
alkoxycarbonyl, aryl, -0-aryl, -S02-aryl,
heteroaryl, or -SO;-heteroaryl, wherein said aryl
and heteroaryl groups may be substituted with 0-4
groups selected from hydrogen, halogen, CF3, Cz-C3
alkyl, or C1-C3 alkoxy;
R20 is selected from:
hydroxy;
Cl to Clo alkoxy;
methylcarbonyloxymethoxy-;
ethylcarbonyloxymethoxy-;
t-butylcarbonyloxymethoxy-;
cyclohexylcarbonyloxymethoxy-;
1-(methylcarbonyloxy)ethoxy-;
1-(ethylcarbonyloxy)ethoxy-;
1-(t-butylcarbonyloxy)ethoxy-;
1-(cyclohexylcarbonyloxy)ethoxy-;
i-propyloxycarbonyloxymethoxy-;
t-butyloxycarbonyloxymethoxy-;
1-(i-propyloxycarbonyloxy)ethoxy-;
1-(cyclohexyloxycarbonyloxy)ethoxy-;
1-(t-butyloxycarbonyloxy)ethoxy-;
dimethylaminoethoxy-;
diethylaminoethoxy-;
(5-methyl-1,3-dioxacyclopenten-2-on-4-yl)methoxy-;
-34-
CA 02221980 1997-11-24
W O 96/37492 PCT/US96/07646
(5-(t-butyl)-1,3-dioxacyclopenten-2-on-4-
yl)methoxy-;
(1,3-dioxa-5-phenyl-cyclopenten-2-on-4-yl)methoxy-
or;
1-(2-(2-methoxypropyl)carbonyloxy)ethoxy-;
R21 is selected from C1-Cg alkyl, C2-C6 alkenyl, C3-C11
cycloalkyl, C4-C11 cycloalkylmethyl, C6-Clo aryl,
C7-C11 arylalkyl, or C1-Clp alkyl substituted with
0-2 R4;
m is 0-2;
n is 0-4;
p is 0-2;
q is 0-1; and
r is 0-2;
with the following provisos:
(1) when b is a double bond, Q and U are not
- ( CH2 ) - ; and
(2) n, m and q are chosen such that the number of
atoms connecting R1 and Y is in the range of
8-14; and
(3) when V is -(phenyl)-Q-, then either: U is not a
direct bond (i.e., U is not -(CH2)n- where n
0) or Q is not a direct bond (i.e., Q is not
-(CH2)n- where n = 0).
[4J Still further preferred are compounds of the above
invention of Formula IIIa, IIIb or IIIc:
-35-
CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
R9
H
R~ ~Uw \ q COR20
R8 ziza
N
.~ O 0
R9
R~ q COR20
O~N 0 R8 zaxb
R9
H
Ri'UN-V A N COR20
N!O 0 Ra rxic
including enantiomeric or diasteriomeric forms thereof,
or mixtures of enantiomeric or diasteriomeric forms
thereof, or pharmaceutically acceptable salt or prodrug
forms thereof wherein:
Rl-U taken together are selected from:
-36-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
R2 N
~i J
NN N N I `H~ H H H+ R3 N
H
R2 N
>-H~ ~--H
+
R3* H
-' ~ ~
H
cx>--1 \
H H H
N O CXN
N O `-H-~NH
IN
H H
R2 N
O
~ \
R3 H H H2N N
N ` ~
I H2N ` or a~N H2N ~,N H
R2 and R3 are independently selected from: H, Cl-C4
alkoxy, halogen, C1-C6 alkyl, or C3-C6 alkenyl;
V is selected from:
- (CHZ)n-,
-(C1-C6 alkylene)-Q-, substituted with 0-1 groups
independently selected from R13 or,
-(C.--C7 alkenylene)-Q-, substituted with 0-1 groups
independently selected from R13, or
-(phenyl)-Q-, said phenyl substituted with 0-i
groups independently selected from R13,
~ Q is selected from:
-37-
CA 02221980 1997-11-24
WO 96/37492 PCT/1JS96/07646
- (CH2)n-,
-0-,
-N(R12) -,
-N (Rlo) C (=O) - , or
-C(=O)N(Rl0) -;
R7 is selected 'from:
H, hydroxy, C1-C4alkoxy, C1-C4 alkylcarbonyl,
-N(Rlo) (Rll) , C0yR18a, gO2N(Ri0) R11, or ORlo;
R8 is selected from:
H, CONR17R18a,
C1-C10 alkyl, C2-C10 alkenyl,
C2-C10 alkynyl, C3-C8 cycloalkyl, pyridinyl, or
aryl, wherein said aryl or pyridinyl groups
are optionally substituted with 0-3
substituents selected from the group
consisting of: C1-C4 alkyl, Cl-C4 alkoxy,
aryl, halo, cyano, CF3, and NO2.
R9 is selected from: H or -NHR16;
R10 is selected from H or C1-C10 alkyl;
R11 is selected from hydrogen, hydroxy, C1 to C8 alkyl,
C3-C6 alkenyl, C3 to C11 cycloalkyl, C4 to C11
cycloalkylmethyl, C1-C6 alkoxy, benzyloxy, C6 to C10
aryl, heteroaryl, heteroarylalkyl, C7 to Cil
arylalkyl, or adamantylmethyl;
R13 is selected from: H, hydroxy, C1-C10 alkoxy,
N(R1o)R11, -11(R16)R17, C1-C1o alkyl substituted with
0-2 R7, aryl substituted with 0-3 R7, heteroaryl
substituted with 0-2 R7, or C1-C6 alkylcarbonyl;
R16 is selected from: -C(=O)-O-R18a,
-38-
CA 02221980 1997-11-24
W O 96/37492 PCT/US96/07646
-SOZ-RiBa or,
-SO2-NHRi8a
R18a is selected from: C1-C8 alkyl, C3-C11 cycloalkyl,
aryl(C1-C6 alkyl)-, (Cz-C6 alkyl)aryl,
7leteroaryl(CI-C6 alkyl)-, (Cj.-C6 alkyl)heteroaryl,
biaryl(C1'C6 alkyl), heteroaryl, or aryl, wherein
said aryl or heteroaryl groups are optionally
substituted with 0-2 R19;
R19 is selected from: H, Br, F, Cl, CF3, CN, NO2, ISHR11,
C1-C4 alkyl, aryl, aryl(C1-C4 alkyl)-, CI-C4 alkoxy,
C1-C4 alkoxycarbonyl, or -0-aryl, wherein said aryl
groups are optionally substituted with 0-3
substituents selected from a group consisting of
halogen, CF3, C1-C3 alkyl, or C1-C3 alkoxy;
R20 is selected from:
hydroxy;
C1 to Clp alkoxy;
methylcarbonyloxymethoxy-;
ethylcarbonylox7ymethoxy-;
t-butylcarbonyloxymethoxy-;
cyclohexylcarbonyloxymethoxy-;
1-(methylcarbonyloxy)ethoxy-;
1-(ethylcarbonyloxy)ethoxy-;
1-(t-butylcarbonyloxy)ethoxy-;
1-(cyclohexylcarbonyloxy)ethoxy-;
i-propyloxycarbonyloxymethoxy-;
t-butyloxycarbonyloxymethoxy-;
1-(i-propyloxycarbonyloxy)ethoxy-;
1-(cyclohexyloxycarbonyloxy)ethoxy-;
1-(t-butyloxycarbonyloxy)ethoxy-;
dimethylaminoethoxy-;
~ 35 diethylaminoethoxy-;
-39-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
(5-methyl-1,3-dioxacyclopenten-2-on-4-yl)methoxy-;
(5-(t-butyl)-1,3-dioxacyclopenten-2-on-4-
yl)methoxy-;
(1,3-dioxa-5-phenyl-cyclopenten-2-on-4-yl)methoxy-
or;
1-(2-(2-methoxypropyl)carbonyloxy)ethoxy-;
n is 0-4; q is 0-1;
with the proviso that n, and q are chosen such that the
number of atoms connecting RI and COR20 is in the range
of 8-14;
[5] Specifically preferred compounds of the above
invention are compounds of Formula I, including
enantiomeric or diasteriomeric forms thereof, or
mixtures of enantiomeric or diasteriomeric forms
thereof, or pharmaceutically acceptable salt or prodrug
forms thereof selected from the group consisting of:
3- [3- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-5-
ylmethylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3- [3- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-5-
ylmethylcarbonylamino]-2-(n-
butyloxycarbonyl-amino)propionic acid,
3- [3- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-5-
ylmethylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid, 35 3- [3- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-5-
-40-
_
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
ylmethylcarbonylamino]-2-(n-
butylsulfonylamino)-propionic acid,
3-[3-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-5-ylmethylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-5-ylmethylcarbonyl amino]-2-
(n-butyloxycarbonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-5-ylmethylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-5-ylmethylcarbonylamino]-2-(n-
butylsulfonyl)aminopropionic acid,
3- [3- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-5-
ylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3- [3- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-5-
ylcarbonylamino]-2-(n-butyloxycarbonyl-
amino)propionic acid,
3- [3- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-5-
ylcarbonylamino]-2-(phenylsulfonylamino)-
propionic acid,
3- [3- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-5-
ylcarbonylamino]-2-(n-butylsulfonylamino)
-propionic acid,
3- [3- [4- (tetrahydropyrimid-2-ylamino)butyl] -
isoxazolin-5-ylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3- [3 - [4 - ( tetrahydropyrimid- 2 -ylamino) butyl] -
isoxazolin-5-ylcarbonyl amino]-2-(n-
butyloxycarbonylamino)propionic acid,
-41-
CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
3- [3- [4- (tetrahydropyrimid-2-ylamino)butyl] -
isoxazolin-5-ylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3-[3-[4-(tetrahydropyrimid-2-ylamino)butyl]-
isoxazolin-5-ylcarbonylamino]-2-(n-
butylsulfonyl)aminopropionic acid,
3- [3- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-5-
ylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3- [3- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-5-
ylcarbonylamino]-2-(phenylsulfonylamino)
propionic acid,
3-[3-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-5-ylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-5-ylcarbonylamino]-2-
(phenylsulf onylamino)propionic acid,
3- [3- [3- (2-aminothiazol-4-
yl)propyl]isoxazolin-5-ylcarbonylamino]-
2-(phenylsulfonylamino) propionic acid,
3- [3- [3- (2-aminothiazol-4-
yl)propyl]isoxazolin-5-ylcarbonylamino]-
2-(benzyloxycarbonylamino)propionic acid,
3- [3- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-5-
ylcarbonylamino]-2-((2,4,6-
trimethylphenyl)sulfonylamino)propionic
acid,
3- [3- [4- (tetrahydropyrimid-2-ylamino)butyl] -
isoxazolin-5-ylcarbonylamino]-2-((2,4,6-
trimethylphenyl)sulfonylamino)propionic
acid,
-42-
CA 02221980 1997-11-24
WO 96/37492 P1T/US96/07646
3-[3-[4-(imidazol-2-ylamino)butylIisoxazolin-
5-ylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3-[3-[4-(imidazol-2-ylamino)butyl]isoxazolin-
5-ylcarbonylamino]-2-
; (phenylsulfonylamino)-propionic acid,
3 - [3 - [4 - ('imidazol - 2 -ylamino) butyll isoxazolin-
5-ylcarbonylamino]-2-
((2,6,dichlorophenyl)-
sulfonylamino)propionic acid,
3-[3-[4-(imidazol-2-ylamino)butyllisoxazolin-
5-ylcarbonylamino]-2-
((2,4,6,trimethyiphenyl)suZfonylamino)
propionic acid,
3- [3- [4- (imidazol-2-ylamino)butyl]isoxazolin-
5-ylcarbonylaminol-2-((4-
biphenyl)sulfonyl- amino)propi.onic acid,
3- [3- [4- (imidazol-2-ylamino)butyl] isoxazolin-
5-ylcarbonylaminol-2-(1-
naphthylsulfonylamino)propionic acid,
3-[3-[3-(imidazol-2-ylamino)propyl]isoxazolin-
5-ylmethylcarbonylamino]-2-
benzyloxycarbonylamino)propionic acid,
3- [3 - [3 - ( imidazol - 2 -ylamino) propyl] isoxazolin-
5-ylmethylcarbonylaminol-2-
(phenylsulf onylamino)propionic acid,
3 - [3 - [3 - ( imidazol - 2 -ylamino) propyl] isoxazolin-
5-ylmethylcarbonylamino]-2-
((2,6,dichlorophenyl)sulfonylamino)propio
nic acid,
3-[3-[3-(imidazol-2-ylamino)propyl]isoxazolin-
5-ylmethylcarbonylamino]-2-
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
-43-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
3-[3-[3-(imidazol-2-ylamino)propyllisoxazolin-
5-ylmethylcarbonylamino]-2-((4-biphenyl)-
sulfonylamino)propionic acid,
3-[3-[3-(imidazol-2-ylamino)propyl]isoxazolin-
5-ylmethylcarbonylamino]-2-(1-
naphthylsulfonylamino)propionic acid,
3- [3- [2- (imidazol-2-ylamino) ethyl] isoxazoli.n-
5-ylcarbonylaminol-2-
(benzyloxycarbonylamino)-propionic acid,
3- [3- [2- (imidazol-2-ylamino) ethyl] isoxazolin-
5-ylcarbonylaminol-2-
(phenylsulfonylamino)-propionic acid,
3- [3- [2- (imidazol-2-ylamino) ethyl] isoxazolin-
5-ylcarbonylamino]-2-
( ( 2 , 6 , dichl.orophenyl ) -
sulfonylamino)propionic acid,
3-[3-[2-(imidazol-2-ylamino)ethyl]isoxazolin-
5-ylcarbonylaminoI-2-
((2,4,6,trimethylphenyl)sulfonylamino)
propionic acid,
3- [3- [2- (imidazol-2-ylamino) ethyl] isoxazolin-
5-ylcarbonylamino]-2-((4-
biphenyl)sulfonyl- amino)propionic acid,
3- [3- [2- (imidazol-2-ylamino) ethyl] isoxazolin-
5-ylcarbonylamino]-2-(1-
naphthylsulfonylamino)propionic acid,
3- [3 - [3 - ( imidazol - 2 -ylaminocarbonyl ) propyl]
isoxazolin-5-ylcarbonylamino]-2-
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
3- [3- [3- (2-aminoimidazol-4-
yl)propyl7isoxazolin-5-ylcarbonylamino]-
2-((2,4,6,trimethylphenyl)sulfonylamino)
propionic acid,
3- [3- [2- (2-aminoimidazol-4-
yl)ethyl]isoxazolin-5-
-44-
CA 02221980 1997-11-24
W O 96/37492 PCT/1JS96/07646
ylmethylcarbornylamino] - 2 -
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
3- [3- [4- (benzimidazol-2-ylamino)butyl]
isoxazolin-5-ylcarbonylaminol-2-
((2,4,6,trimethylphenyl)sulfonylamino)
propi=onic acid,
3-.[3- [3- (benzimidazol-2-ylamino)propyl]
isoxazolin-5-ylmethylcarbonylamino]-2-
((2,4,6,trimethylphenyl)sulfonylamino)
propionic acid,
3- [3- [3- (benzimidazol-2-
ylaminocarbonyl)propyl] isoxazolin-5-
ylcarbonylaminol-2-
((2,4,6,trimethyiphenyl)sulfonylamino)-
propionic acid,
3- [3- [4- (4-methylimidazol-2-ylamino)butyl]-
isoxazolin-5-ylcarbonylaminoI-2-
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
.3- [3- [3- (4-methylimidazol-2-ylamino)propyl] -
isoxazolin-5-ylmethylcarbonylaminol-2-
((2,4,6,trimethyiphenyl)sulfonylamino)-
propionic acid,
3- [3- [4- (4, 5-dimethylimidazol-2-
ylamino)butyl]- isoxazolin-5-
ylcarbonylamino]-2-
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
3- [3- [3- (4, 5-dimethylimidazol-2-
ylamino)propyll- isoxazolin-5-
ylmethylcarbonylamino]-2-
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
3-[3-[3-(4,5-dimethylimidazol-2-
ylaminocarbonyl)propyl] isoxazolin-5-
-45-
CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
ylcarbonylamino]-2-
((2,4,6,trimethyiphenyl)sulfonylamino)-
propionic acid,
3-[3-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]isoxazolin-5-
ylcarbonylamino] -2-
((2,4,6,trimethyiphenyl)sulfonylamino)-
propionic acid, 3-[3-[3-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)propyl]isoxazolin-
5-ylmethylcarbonylamino]-2-
((2,4,6,trimethyiphenyl)sulfonyl-
amino)propionic acid,
3-[3-[4-(pyridin-2-ylamino)butyl]isoxazolin-5-
ylcarbonylamino]-2-
((2,4,6,trimethylphenyl)
sulfonylamino)propionic acid,
3 - [3 - [3 - (pyridin- 2 -ylamino) propyll isoxazolin-
5-ylmethylcarbonylaminol-2-
((2,4,6,trimethylphenyl)sulfonyl-
amino)propionic acid,
3- [3- [3- (2-pyridin-6-yl)propyl] isoxazolin-5-
ylcarbonylamino]-2-
((2,4,6,trimethylphenyl)
sulfonylamino)propionic acid,
3- [3- [2- (2-aminopyridin-6-yl) ethyl] isoxazolin-
5-ylmethylcarbonylaminol-2-
((2,4,6,trimethylphenyl)sulfonylamino)-
propionic acid,
3-[3-[3-(7-azabenzimidazol-2-yl)propyl]
isoxazolin-5-ylcarbonylamino]-2-
((2,4,6,trimethyiphenyl)sulfonylamino)-
propionic acid,
3-[5-[3-(imidazolin-2-ylamino)propyl]
isoxazolin-3-ylmethylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
-46-
CA 02221980 1997-11-24
WO 96/37492 PCT/1JS96/07646
3- [5- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-3-
ylmethylcarbonylamino]-2-(n-
butyloxycarbonyl-amino)propionic acid,
3- [5- [3- (imidazolin-2-
_ ylamino)propyl]isoxazolin-3-
ylmethylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3- [5- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-3-
ylmethylcarbonylamino]-2-(2,4,6-
trimethylphenylsulfonylamino)propionic
acid,
3- [5- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-3-
ylmethylcarbonylaminol-2-(n-
butylsulfonylamino)-propionic acid,
3- [5- [3- (tetrahydropyrimid-2-ylamino)propyl] -
isoxazolin-3-ylmethylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylmethylcarbonyl amino]-2-
(n-butyloxycarbonylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylmethylcarbonylamino]-2-
(phenylsulf onylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylmethylcarbonylamino]-2-
(2,4,6-trimethylphenylsulfonylamino)
propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylmethylcarbonylaminol-2-(n-
butylsulfonyl)aminopropionic acid,
3- [5- [4- (imidazolin-2-yl
amino)butyl]isoxazolin-3-
-47-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
ylcarbonylamino]-2-
(benzyloxycarbonylamino)- propionic acid,
3- [5- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-3- =
ylcarbonylamino]-2-(n-butyloxycarbonyl-
amino)propionic acid,
3- [5- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3- [5- [4- (imidazolin-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-(n-butylsulfonylamino)
propionic acid,
3-[5-[4-(tetrahydropyrimid-2-ylamino)butyl]-
isoxazolin-3-ylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3- [5- [4- (tetrahydropyrimid-2-ylami.no)butyl] -
isoxazolin-3-ylcarbonyl amino]-2-(n-
butyloxycarbonylamino)propionic acid,
3-[5-[4-(tetrahydropyrimid-2-ylamino)butyl]-
isoxazolin-3-ylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3-[5-[4-(tetrahydropyrimid-2-ylamino)butyl]-
isoxazolin-3-ylcarbonylamino]-2-(n-
butylsulfonyl)aminopropionic acid,
3- [5- [3- (imidazol-2-yl
amino)propyl]isoxazolin-3-
ylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3- [5- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-3-
ylcarbonylamino]-2-(n-propyloxycarbonyl-
amino)propionic acid, w
3- [5- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-3-
-48-
CA 02221980 1997-11-24
W 96/37492 PCT/US96/07646
ylcarbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3- [5- [3- (imidazolin-2-
ylamino)propyl]isoxazolin-3-
ylcarbonylamino] - 2 - (n-
propylsulfonylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylmethylcarbonyl amino]-2-
(n-propyloxycarbonylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylcarbonylamino]-2-
(phenylsulf onylamino)propionic acid,
3-[5-[3-(tetrahydropyrimid-2-ylamino)propyl]-
isoxazolin-3-ylcarbonylamino]-2-(n-
propylsulfonyl)aminopropionic acid,
3- [5- [2- (imidazolin-2-
ylamino)ethyl]isoxazolin-3-
ylcarbonylamino]-2-(phenylsulfonylamino)-
propionic acid,
3-[5-[4-(pyridin-2-ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-(phenylsulfonylamino)-
propionic acid,
3- [5- [4- (pyridin-2-ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-(2,6-dichiorophenyl-
sulfonylamino)propionic acid,
3- [5- [4- (pyridin-2-ylamino)butyl] isoxazolin-3-
ylcarbonylamino]-2-(2,4,6-
trimethyiphenyl- sulfonylamino)propionic
acid,
3-[5-[4-(pyridin-2-ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-(4-
biphenylsulfonylamino)propionic'acid,
-49-
CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
3 - [5 - [4 - (pyridin- 2 -ylamino) butyl] isoxazolin- 3 -
ylcarbonylamino]-2-(1-naphthylsulfonyl-
amino)propionic acid,
3- [ 5 - [ 3 - ( 2 - aminopyridin - 6 -
yl)propyl]isoxazolin-3-ylcarbonylamino]-
2-(2,4,6-trimethylphenyl
sulfonylamino)propionic acid,
3- [5- [4- (imidazol-2-ylamino)butyl] isoxazolin-
3-ylcarbonylamino]-2-
(phenylsulfonylamino)- propionic acid,
3-[5-[4-(imidazol-2-ylamino)butyl]isoxazolin-
3-ylcarbonylamino]-2-(2,6-dichiorophenyl-
sulfonylamino)propionic acid,
3- [5- [4- (imidazol-2-ylama.no)butyl] isoxazolin-
3-ylcarbonylaminol-2-(2,4,6-
trimethylphenyl- sulfonylamino)propionic
acid,
3 - [ 5 - [4 - ( imidazol - 2 -ylamino) butyl] isoxazolin-
3-ylcarbonylaminol-2-(4-biphenylsulfonyl-
amino)propionic acid,
3- [5- [4- (imidazol-2-ylamino)butyl] isoxazolin-
3-ylcarbonylamino]-2-(1-naphthylsulfonyl-
amino)propionic acid,
3- [5- [3- (2-aminoimidazol-4-
yl)propyl]isoxazolin-3-ylcarbonylamino]-
2-(phenylsulfonylamino)- propionic acid,
3- [5- [3- (2-aminoimidazol-4-
yl)propyl]isoxazolin-3-ylcarbonylamino]-
2-(2,6-dichiorophenyl-
sulfonylamino)propionic acid,
3- [5- [3- (2-aminoimidazol-4-
yl)propyl]isoxazolin-3-ylcarbonylamino]-
2-(2,4,6-trimethylphenyl
sulfonylamino)propionic acid,
3- [5- [3- (2-aminoimi.dazol-4-
yl)propyl]isoxazolin-3-ylcarbonylamino]-
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2-(4-biphenylsulfonylamino)propionic
acid,
3- [5- [3- (2-aminoimidazol-4-
yl)propyllisoxazolin-3-ylcarbonylamino]-
2-(1-naphthylsulfonylamino)propionic
acid,
3- [5- [2- (=imidazol-2-ylamino) eth~,,l] isoxazolin-
3-ylcarbonylaminol-2-
(phenylsulfonylamino)- propionic acid,
3-[5 -[2 -( imidazol- 2-ylamino) ethyll isoxazolin-
3-ylcarbonylamino]-2-(2,6-dichlorophenyl-
sulfonylamino)propionic acid,
3- [5- [2- (imidazol-2-ylamino) ethyll isoxazolin-
3-ylcarbonylaminol-2-(2,4,6-
trimethylphenyl- sulfonylamino)propionic
acid,
3- [5- [2- (imidazol-2-ylamino) ethyl3 isoxazolin-
3-ylcarbonylaminol-2-(4-biphenylsulfonyl-
amino)propionic acid,
3- [5- [2- (imidazol-2-ylamino) ethyl] isoxazolin-
3-ylcarbonylamino]-2-(1-naphthylsulfonyl-
amino)propionic acid,
3-[5-[3-(imidazol-2-ylaminocarbonyl)propyl]
isoxazolin-3-ylcarbonylamino]-2-(2,4,6-
trimethylphenylsulfonylamino) propionic
acid,
3-[5-[3-(benzimidazol-2-ylaminocarbonyl)
propyllisoxazolin-3-ylcarbonylamino]-2-
(2,4,6-trimethylphenylsulfonylamino)
propionic acid,
3- [5- [4- (benzimidazol-2-ylamino)butyl]
isoxazolin-3-ylcarbonylamino]-2-
(phenyisulfonylamino)propionic acid,
3-[5-[4-(benzimidazol-2-ylamino)butyl]
isoxazolin-3-ylcarbonylamino]-2-(2,6-
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dichlorophenylsulfonylamino)propionic
acid,
3- [5- [4- (benzimidazol-2-~.rlamino)butyl]
isoxazolin-3-ylcarbonylamino]-2-(2,4,6- 5
trimethylphenylsulfonylamino)propionic
acid,
3- [5- [4- (benzimidazol-2-ylamino)butyl]
isoxazolin-3-ylcarbonylaminol-2-(4-
biphenylsulfonylamino)propionic acid,
3- [5- [4- (benzimidazol-2-ylamino)butyl]
isoxazolin-3-ylcarbonylaminol-2-(1-
naphthylsulfonylamino)propionic acid,
3- [5- [4- (4-methylimidazol-2-ylamino)butyl] -
isoxazolin-3-ylcarbonylamino]-2-
(phenylsulfonylamino) propionic acid,
3- [5- [4- (4-methylimidazol-2-ylamino)butyl] -
isoxazolin-3-ylcarbonylaminol-2-(2,6-
dichlorophenylsulfonylamino)propionic
acid,
3- [5- [4- (4-methylimidazol-2-ylamino)butyll -
isoxazolin-3-ylcarbonylaminol-2-(2,4,6-
trimethyiphenylsulfonylamino)propionic
acid,
3- [5- [4- (4-methylimidazol-2-ylamino)butyl] -
isoxazolin-3-ylcarbonylamino]-2-(4-
biphenylsulfonylamino)propionic acid,
3- [5- [4- (4-methylimidazol-2-ylamino)butyl] -
isoxazolin-3-ylcarbonylamino]-2-(1-
naphthylsulfonylamino)propionic acid,
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyll-isoxazolin-3-
ylcarbonylamino]-2-(phenylsulfonylamino)
propionic acid,
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyl]-isoxazolin-3-
ylcarbonylaminol-2-(2,6-
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dichlorophenylsulfonylamino)propionic
acid,
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyl]-isoxazolin-3-
ylcarbonylamino]-2-(2,4,6-
_ trimethylphenylsulfonylamino)propionic
acid;
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyl]-isoxazolin-3-
ylcarbonylamino]-2-(4-
biphenylsulfon~rlamino)propionic acid,
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyl]-isoxazolin-3-
ylcarbonylamino]-2-(1-
naphthylsulfonylamino)propionic acid,
3-[5-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyll-isoxazolin-3-
ylcarbonylamino]-2-(phenylsulfonylamino)
propionic acid,
3-[5-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]-isoxazolin-3-
ylcarbonylamino]-2-(2,6-dichlorophenyl-
sulfonylamino)propionic acid,
3-[5-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]-isoxazolin-3-yl-
carbonylamino]-2-(2,4,6-trimethylphenyl-
sulfonylamino)propionic acid,
3-[5-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-(4-biphenyl-
sulfonylamino)propionic acid,
3-[5-[4-(4-,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]-isoxazolin-3-
ylcarbonylamino]-2-(1-naphthyl-
sulfonylamino)propionic acid,
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3- [5- [3- (7-azabenzimidazol-2-yl)propyl]
isoxazolin-3-ylcarbonylamino]-2-(2,4,6-
trimethylphenylsulfonylamino) propionic
acid,
3- [5- [4- (imidazol-2-ylamino)butyl] isoxazolin-
3-ylcarbonylamino]-2-[(2,6-dimethyl-4- =
phenyl) phenylsulfonylamino] propionic
acid,
3- [5- [4- (4-methylimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-[(2,6-dimethyl-4-
phenyl)phenylsulfonylamino]propionic
acid,
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino7-2-[(2,6-dimethyl-4-
phenyl)phenylsulfonylamino]propionic
acid,
3-[5-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-[(2,6-dimethyl-4-
phenyl)phenylsulfonylamino]propionic
acid,
3- [5- [4- (imidazol-2-ylamino)butyl] isoxazolin-
3-ylcarbonylamino]-2-[(2,6-dichloro-4-
phenyl)phenylsulfonylamino]propionic
acid,
3- [5- [4- (4-methylimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-[(2,6-dichloro-4-
phenyl)phenylsulfonylamino]propionic
acid,
3-[5-[4-(4,5-dimethylimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-[(2,6-dichloro-4-
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phenyl)phenylsulfonylamino]propionic
acid,
3-[5-[4-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)butyl]isoxazolin-3-
ylcarbonylamino]-2-[(2,6-dichloro-4-
phenyl) phenylsulfonylamino] propionic
acid,'
3--[5- [4- (imidazol-2-ylamino)butyl] isoxazolin-
3-ylcarbonylamino]-3-
(phenylsulfonylmethyl) propionic acid,
3-[5-[4-(imidazol-2-ylamino)butyl]isoxazolin-
3-ylcarbonylamino]-3-(1-adamantylmethyl-
aminocarbonyl)propionic acid,
3-[5-[4-(imidazol-2-ylamino)butyl]isoxazolin-
3-ylcarbonylaminol-3-(3-
pyridinyl)propionic acid,
3- [3- [3- (imidazolin-2-yl amino)propyloxy]
isoxazol-5-ylcarbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3-[3-[3-(imidazolin-2-ylamino)propyloxy]
isoxazol-5-yl carbonyl amino]-2-(n-
butyloxycarbonyl-amino)propionic acid,
3- [3- [3- (imidazolin-2-ylamino)propyloxy]
isoxazol-5-yl carbonylamino]-2-
(phenylsulfonylamino)propionic acid,
3-[3-[3-(imidazolin-2-ylamino)propyloxy]
isoxazol-5-yl carbonyl amino]-2-(n-
butylsulfonylamino)-propionic acid,
3- [3 - [ 3 - ( tetrahydropyrimid- 2 -
ylamino)propyloxy]-isoxazol-5-
ylcarbonylamino]-2-
(benzyloxycarbonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-
ylamino)propyloxy]-isoxazol-5-ylcarbonyl
amino]-2-(n-
butyloxycarbonylamino)propionic acid,
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3-[3-[3-(tetrahydropyrimid-2-
ylamino)propyloxy]-isoxazol-5-
ylcarbonylaminol-2-
'(phenylsulfonylamino)propionic acid,
3 - [3 - [3 - ( tetrahydropyrimid- 2 -
ylamino)propyloxy]-isoxazol-5-yl
carbonylaminol - 2 - (n-
butylsulfonyl)aminopropionic acid,
3-[3-[2-(imidazolin-2-yl amino)ethyloxy]
isoxazol-5-yl carbonylamino]-2-
(benzyloxycarbonylamino)-propionic acid,
3- [3- [3- (imidazolin-2-
ylamino)ethyloxy]isoxazol-5-yl
carbonylaminol-2-(n-butyloxycarbonyl-
amino)propionic acid,
3- [3- [3- (imidazolin-2-
ylamino)ethyloxy]isoxazol-5-yl
carbonylaminol-2-(phenylsulfonylamino)
propionic acid,
3- [3- [3- (imidazolin-2-
ylamino)ethyloxylisoxazol-5-
ylcarbonylamino]-2-(n-butylsulfonylamino)
propionic acid,
3-[3-[3-(tetrahydropyrimid-2-yl
amino)ethyloxy]-isoxazol-5-
ylcarbonylaminol-2-
(benzyloxycarbonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-
ylamino)ethyloxy]-isoxazol-5-
ylcarbonylaminol-2-(n-
butyloxycarbonylamino)propionic acid,
3-[3-[3-(tetrahydropyrimid-2-
ylamino)ethyloxy]-isoxazol-5-
ylcarbonylamino]-2- 35 (phenylsulfonylamino)propionic=acid,
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3-[3-[3-(tetrahydropyrimid-2-
ylamino)ethyloxy]-isoxazol-5-
ylcarbonylamino]-2-(n-
butylsulfonylamino)propionic acid,
3- [3- [3- (imidazol-2-ylamino)propyloxy] -
isoxazol-5-ylcarbonylamino]-2-
(phenylsulf ornylamino) propionic acid,
3 - [3 - [3 - (benzimidazol- 2 -ylamino ) propyloxy] -
isoxazol-5-ylcarbonylamino]-2-
(phenylsulfonylamino) propionic acid,
3-[3-[3-(4-methylimidazol-2-
ylamino)propyloxyI-isoxazol-5-
ylcarbonylamino]-2-(phenylsulfonylamino)
propionic acid,
3-[3-[3-(4,5-dimethylimidazol-2-
ylamir_o ) prop,-loxy] - isoxazol - 5 -
ylcarbonylamino]-2-(phenylsulfonylamino)
propionic acid,
3-[3-[3-(4,5,6,7-tetrahydrobenzimidazol-2-
ylamino)propyloxy]-isoxazol-5-
ylcarbonylamino]-2-(phen_vlsulfonylamino)
propionic acid,
3 - [3 - [3 - (pyridin- 2 -ylamino) propY,loxy] -
isoxazol-5-ylcarbonylamino]-2-
(phenyisulfonylainino) propionic acid,
3 - [3 - [3 - ( imidazol - 2 -ylaminocarbonyl) ethoxy] -
isoxazol-5-ylcarbonylamino]-2-
(phenyisulfonylamino) propionic acid,
In the present invention it has been discovered
that the compounds of Formula I above are useful as
inhibitors of cell-matrix and cell-cell adhesion
processes. The present invention includes novel
compounds of Formula I and methods for using such
compounds for the prevention or treatment of diseases
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resulting from abnormal cell adhesion to the
extracellular matrix which comprises administering to a
host in need of such treatment a therapeutically
effective amount of such compound of Formula I. 5 In the present invention it
has also been
discovered that the compounds of Formula I above are
useful as inhibitors of avP3. The compounds of the
present invention inhibit tYie binding of vitronectin to
avP3 and inhibit cell adhesion.
The present invention also provides pharmaceutical
compositions comprising a compound of Formula I and a
pharmaceutically acceptable carrier.
The compounds of Formula I of the present invention
are useful for the treatment (including prevention) of
angiogenic disorders. The term "angiogenic disorders"
as used herein includes conditions involving abnormal
neovascularization, such as tumor metastasis and ocular
neovascularization, including, for example, diabetic
retinopathy, neovascular glaucoma, age-related macular
degeneration, and retinal vein occlusion, comprising
administering to a mammal in need of such treatment a
therapeutically effective amount of a compound of
Formula I described above.
The compounds of Formula I of the present invention
may be useful for the treatment or prevention of other
diseases which involve cell adhesion processes,
including, but not limited to, inflammation, bone
degradation, thromboembolic disorders, restenosis,
rheumatoid arthritis, asthma, allergies, adult
respiratory distress syndrome, graft versus host
disease, organ transplantation rejection, septic shock,
psoriasis, eczema, contact dermatitis, osteoporosis,
osteoarthritis, atherosclerosis, inflammatory bowel
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disease and other autoimznune diseases. The compounds of
Formula I of the present invention may also be useful
for wound healing.
The term "thromboembolic disorders" as used herein
includes conditions involving platelet activation and
aggregation, such as arterial or venous cardiovascular
or cerebrovascular thromboembolic disorders, including,
for example, thrombosis, unstable angina, first or
recurrent myocardial infarction, ischemic sudden death,
transient ischemic attack, stroke, atherosclerosis,
venous thrombosis, deep vein thrombosis,
thrombophiebitis, arterial embolism, coronary and
cerebral arterial thrombosis, myocardial infarction,
cerebral embolism, kidney embolisms, pulmonary
embolisms, or such disorders associated with diabetes,
comprising administering to a mammal in need of such
treatment a therapeutically effective amount of a
compound of Formula I described above.
The compounds of the present invention may be used
for other ex v.ivo applications to prevent cellular
adhesion in biological samples.
The compounds of the present invention can also be
administered in combination with one or more additional
therapeutic agents selected from: anti-coagulant or
coagulation inhibitory agents, such as heparin or
warfarin; anti-platelet or platelet inhibitory agents,
such as aspirin, piroxicam, or ticlopidine; thrombin
inhibitors such as boropeptides, hirudin or argatroban;
or thrombolytic or fibrinolytic agents, such as
plasminogen activators, anistreplase, urokinase, or
streptokinase.
The compounds of Formula I of the present
invention can be administered in combination with one or
more of the foregoing additional therapeutic agents,
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thereby to reduce the doses of each drug required to
achieve the desired therapeutic effect. Thus, the
combination treatment of the present invention permits
the use of lower doses of each component, with reduced
adverse, toxic effects of each component. A lower
dosage'minimizes the potential of side effects of the
compounds, thereby providing an increased margin of
safety relative to the margin of safety for each
component when used as a single agent. Such combination
therapies may be employed to achieve synergistic or
additive therapeutic effects for the treatment of
thromboembolic disorders.
By "therapeutically effective amount" it is meant
an amount of a compound of Formula I that when
administered alone or in combination with an additional
therapeutic agent to a cell or mammal is effective to
prevent or ameliorate the thromboembolic disease
condition or the progression of the disease.
By "administered in combination" or "combination
therapy" it is meant that the compound of Formula I and
one or more additional therapeutic agents are
administered concurrently to the mammal being treated.
When administered in combination each component may be
administered at the same time or sequentially in any
order at different points in time. Thus, each component
may be administered separately but sufficiently closely
in time so as to provide the desired therapeutic effect.
The term anti-coagulant agents (or coagulation
inhibitory agents), as used herein, denotes agents that
inhibit blood coagulation. Such agents include warfarin
(available as COUMADINTM) and heparin.
The term anti-platelet agents (or platelet
inhibitory agents), as used herein, denotes agents that inhibit platelet
function such as by inhibiting the
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aggregation, adhesion or granular secretion of
platelets. Such agents include the various known
non-steroidal anti-inflammatory drugs (NSAIDS) such as
aspirin, ibuprofen, naproxen, sulindac, indomethacin,
mefenamate, droxicam, diclofenac, sulfinpyrazone, and
piroxicam, including pharmaceutically acceptable salts
or prodrugs thereof. Of the NSAIDS, aspirin
(acetylsalicyclic acid or ASA), and piroxicam.
Piroxicam is commercially available from Pfizer Inc.
(New York, NY), as FELDANETM. Other suitable anti-
platelet agents include ticlopidine, including
pharmaceutically acceptable salts or prodrugs thereof.
Ticlopidine is also a preferred compound since it is
known to be gentle on the gastro-intestinal tract in
use. Still other suitable platelet inhibitory agents
include thromboxane-A2-receptor antagonists and
thromboxane-A2-synthetase inhibitors, as well as
pharmaceutically acceptable salts or prodrugs thereof.
The phrase thrombin inhibitors (or anti-thrombin
agents), as used herein, denotes inhibitors of the
serine protease thrombin. By inhibiting thrombin,
various thrombin-mediated processes, such as
thrombin-mediated platelet activation (that is, for
example, the aggregation of platelets, and/or the
granular secretion of plasminogen activator inhibitor-1
and/or serotonin) and/or fibrin formation are disrupted.
Such inhibitors include boroarginine derivatives and
boropeptides, hirudin and argatroban, including
pharmaceutically acceptable salts and prodrugs thereof.
Boroarginine derivatives and boropeptides include
N-acetyl and peptide derivatives of boronic acid, such
as C-terminal a-aminoboronic acid derivatives of lysine,
ornithine, arginine, homoarginine and corresponding
isothiouronium analogs thereof. The term hirudin, as
used herein, includes suitable derivatives or analogs of
hirudin, referred to herein as hirulogs, such as
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disulfatohirudin. Boropeptide thrombin inhibitors
include compounds described in Kettner et al., U.S.
Patent No. 5,187,157 and European Patent Application
Publication Number 293 881 A2, the disclosures of which
are hereby incorporated herein by reference. Other
suitable boroarginine derivatives and boropeptide
thrombin inhibitors include those disclosed in PCT
Application Publication Number 92/07869 and European
Patent Application Publication Number 471 651 A2, the
disclosures of which are hereby incorporated herein by
reference, in their entirety.
The phrase thrombolytics (or fibrinolytic) agents
(or thrombolytics or fibrinolytics), as used herein,
denotes agents that lyse blood clots (thrombi). Such
agents include tissue plasminogen activator,
anistreplase, urokinase or streptokinase, including
pharmaceutically acceptable salts or prodrugs thereof.
Tissue plasminogen activator (tPA) is commercially
available from Genentech Inc., South San Francisco,
California. The term anistreplase, as used herein,
refers to anisoylated plasminogen streptokinase
activator complex, as described, for example, in
European Patent Application No. 028,489, the disclosures
of which are hereby incorporated herein by reference
herein, in their entirety. Anistreplase is commercially
available as EMINASETM. The term urokinase, as used
herein, is intended to denote both dual and single chain
urokinase, the latter also being referred to herein as
prourokinase.
Administration of the compounds of Formula I of the
invention in combination with such additional
therapeutic agent, may afford an efficacy advantage over
the compounds and agents alone, and may do so while
permitting the use of lower doses of each. A lower
dosage minimizes the potential of side effects, thereby
providing an increased margin of safety.
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The compounds of the present invention are also
useful as standard or reference compounds, for example
as a quality standard or control, in tests or assays
involving the binding of vitronectin or fibrinogen to
avO3. Such compounds may be provided in a commercial
kit, for example, for use in pharmaceutical research
involving avP3.. The compounds of the present invention
may also be used in diagnostic assays involving avP3.
The compounds herein described may have
asymmetric centers. Unless otherwise indicated, all
chiral, diastereomeric and racemic forms are included in
the present invention. Many geometric isomers of
olefins, C=N double bonds, and the like can also be
present in the compounds described herein, and all such
stable isomers are contemplated in the present
invention. It will be appreciated that compounds of the
present invention that contain asymmetrically
substituted carbon atoms may be isolated in optically
active or racemic forms. It is well known in the art
how to prepare optically active forms, such as by
resolution of racemic forms or by synthesis, from
optically active starting materials. All chiral,
diastereomeric, racemic forms and all geometric isomeric
forms of a structure are intended, unless the specific
stereochemistry or isomer form is specifically
indicated.
When any variable (for example but not limited to,
R2, R4, R6, R7, R8, R12, and R14, n, etc.) occurs more
than one time in any constituent or in any formula, its
definition on each occurrence is independent of its
definition at every other occurrence. Thus, for
example, if a group is shown to be substituted with 0-2
R4, then said group may optionally be substituted with
up to two R4 and R4 at each occurrence is -selected
independently from the defined list of possible R4.
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Also, by way of example, for the group -N(R5a)2, each of
the two R5a substituents on N is independently selected
from the defined list of possible Rsa. Similarly, by
way of example, for the group -C(R7)2-, each of the two
R7 substituents on C is independently selected from the
defindd list of possible R7.
When a bond to a substituent is shown to cross the
bond connecting two atoms in a ring, then such
substituent may be bonded to any atom on the ring. When
a bond joining a substituent to another group is not
specifically shown or the atom in such other group to
which the bond joins is not specifically shown, then
such substituent may form a bond with any atom on such
other group.
When a substituent is listed without indicating the
atom via which such substituent is bonded to the rest of
the compound of Formula I, then such substituent may be
bonded via any atom in such substituent. For example,
when the substituent is piperazinyl, piperidinyl, or
tetrazolyl, unless specified otherwise, said
piperazinyl, piperidinyl, tetrazolyl group may be bonded
to the rest of the compound of Formula I via any atom in
such piperazinyl, piperidinyl, tetrazolyl group.
Combinations of substituents and/or variables
are permissible only if such combinations result in
stable compounds. By stable compound or stable
structure it is meant herein a compound that is
sufficiently robust to survive isolation to a useful
degree of purity from a reaction mixture, and
formulation into an efficacious therapeutic agent.
The term "substituted", as used herein, means
that any one or more hydrogen on the designated atom is
replaced with a selection from the indicated group,
provided that the designated atom's normal valency is
not exceeded, and that the substitution results in a
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stable compound. When a substitent is keto (i.e., =0),
then 2 hydrogens on the atom are replaced.
As used herein, "alkyl is intended to include
both branched and straight-chain saturated aliphatic
hydrocarbon groups having the specified number of carbon
atoms (for example, "C1-Cl011 denotes alkyl having 1 to 10
carbon atoms); "haloalkyl is intended to include both
branched and straight-chain saturated aliphatic
hydrocarbon groups having the specified number of carbon
atoms, substituted with 1 or more halogen (for example
-CvFw where v = 1 to 3 and w = 1 to (2v+1)); "alkoxy
represents an alkyl group of indicated number of carbon
atoms attached through an oxygen bridge; cycloalkyl is
intended to include saturated ring groups, including
mono-,bi- or poly-cyclic ring systems, such as
cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,
cycloheptyl, cyclooctyl, and adama.ntyl; and
"biycloalkyl is intended to include saturated bicyclic
ring groups such as [3.3.0]bicyclooctane,
[4.3..0]bicyclononane, [4.4.0]bicyclodecane (decalin),
[2.2.2]bicyclooctane, and so forth. "Alkenyl" is
intended to include hydrocarbon chains of either a
straight or branched configuration and one or more
unsaturated carbon-carbon bonds which may occur in any
stable point along the chain, such as ethenyl, propenyl
and the like; and "alkynyl is intended to include
hydrocarbon chains of either a straight or branched
configuration and one or more triple carbon-carbon bonds
which may occur in any stable point along the chain,
such as ethynyl, propynyl and the like.
The terms "alkylene", "alkenylene", "phenylene",
and the like, refer to alkyl, alkenyl, and phenyl
groups, respectively, which are connected by two bonds
to the rest of the structure of Formula I. Such
= "alkylene", "alkenylene", "phenylene", and the like, may
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alternatively and equivalently be denoted herein as
"-(alkyl)-", -(alkyenyl)-" and "-(phenyl)-", and the
like.
"Halo" or "halogen" as used herein refers to
fluoro, chloro, bromo and iodo; and "counterion" is used
to represent a small, negatively charged species such as
chloride, bromide, hydroxide, acetate, sulfate and the
like. As used herein, "aryl" or "aromatic residue" is
intended to mean phenyl or naphthyl; the term
"arylalkyl represents an aryl group attached through an
alkyl bridge.
As used herein, "carbocycle" or "carbocyclic
residue" is intended to mean any stable 3- to 7-
membered monocyclic or bicyclic or 7- to 14-membered
bicyclic or tricyclic or an up to 26-membered polycyclic
carbon ring, any of which may be saturated, partially
unsaturated, or aromatic. Examples of such carbocyles
include, but are not limited to, cyclopropyl,
cyclopentyl, cyclohexyl, phenyl, biphenyl, naphthyl,
indanyl, adamantyl, or tetrahydronaphthyl (tetralin).
As used herein, the term "heterocycle" or
"heterocyclic" is intended to mean a stable 5- to 7-
membered monocyclic or bicyclic or 7- to 10-membered
bicyclic heterocyclic ring which may be saturated,
partially unsaturated, or aromatic, and which consists
of carbon atoms and from 1 to 4 heteroatoms
independently selected from the group consisting of N, 0
and S and wherein the nitrogen and $ulfur heteroatoms
may optionally be oxidized, and the nitrogen may
optionally be quaternized, and including any bicyclic
group in which any of the above-defined heterocyclic
rings is fused to a benzene ring. The heterocyclic ring
may be attached to its pendant group at any heteroatom
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W O 96137492 PCT/fJS96107646
or carbon atom which results in a stable structure. The
heterocyclic rings described herein may be substituted
on carbon or on a nitrogen atom if the resulting
compound is stable. Examples of such heterocycles
include, but are not limited to, pyridyl (pyridinyl),
pyrimidinyl, furanyl (furyl), thiazolyl, thienyl,
pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl,
benzofuranyl, benzothiophenyl, indolyl, indolenyl,
isoxazolinyl, isoxazolyl, quinolinyl, isoquinolinyl,
benzimidazolyl, piperidinyl, 4-piperidonyl,
pyrrolidinyl, 2-pyrrolidonyl, pyrrolinyl,
tetrahydrofuranyl, tetrahydrocduinolinyl,
tetrahydroisoquinolinyl, decahydroquinolinyl or
octahydroisoquinolinyl, azocinyl, triazinyl, 6H-1,2,5-
thiadiazinyl, 2H,6H-1,5,2-dithiazinyl, thianthrenyl,
pyranyl, isobenzofuranyl, chromenyl, xanthenyl,
phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl,
pyrazolyl, isothiazolyl, isoxazolinyl, isoxazolyl,
oxazolyl, pyridinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl,
indolyl, 1H-indazolyl, purinyl, 4H-quinolizinyl,
isoquinolinyl, quinolinyl, phthalazinyl, naphthyridinyl,
quinoxalinyl, quinazolinyl, cinnolinyl, pteridinyl,
4aH-carbazole, carbazole, Q-carbolinyl, phenanthridinyl,
acridinyl, perimidinyl, phenanthrolinyl, phenazinyl,
phenarsazinyl, phenothiazinyl, furazanyl, phenoxazinyl,
isochromanyl, chromanyl, pyrrolidinyl, pyrrolinyl,
imidazolidinyl, imidazolinyl, pyrazolidinyl,
pyrazolinyl, piperidinyl, piperazinyl, indolinyl,
isoindolinyl, quinuclidinyl, morpholinyl or
oxazolidinyl. Also included are fused ring and spiro
compounds containing, for example, the above
heterocycles.
At used herein, the term heteroaryl" refers to
aromatic heterocyclic groups. Such heteroaryl groups
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are preferably 5-6 membered monocylic groups or 8-10
membered fused bicyclic groups. Examples of such
heteroaryl groups include, but are not limited to
pyridyl (pyridinyl), pyrimidinyl, furanyl (furyl),
thiazolyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl,
indolyl, isoxazolyl, oxazolyl, pyrazinyl, pyrimidinyl, w
pyridazinyl, benzofuranyl, benzothienyl, benzimidazolyl,
quinolinyl, or isoquinolinyl
As used herein, "pharmaceutically acceptable salts"
refer to derivatives of the disclosed compounds wherein
the parent compound of Formula I is modified by making
acid or base salts of the compound of Formula I.
Examples of pharmaceutically acceptable salts include,
but are not limited to, mineral or organic acid salts of
basic residues such as amines; alkali or organic salts
of acidic residues such as carboxylic acids; and the
like.
"Prodrugs" are considered to be any covalently
bonded carriers which release the active parent drug
according to Formula I in vivo when such prodrug is
administered to a mammalian subject. Prodrugs of the
compounds of Formula I are prepared by modifying
functional groups present in the compounds in such a way
that the modifications are cleaved, either in routine
manipulation or in vivo, to the parent compounds.
Prodrugs include compounds of Formula I wherein
hydroxyl, amino, sulfhydryl, or carboxyl groups are
bonded to any group that, when administered to a
mammalian subject, cleaves to form a free hydroxyl,
amino, sulfhydryl, or carboxyl group respectively.
Examples of prodrugs include, but are not limited to,
acetate, formate and benzoate derivatives of alcohol and
amine functional groups in the compounds of Formula I,
and the like. Examples of representative carboxyl and -68-
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amino prodrugs are included under the definition of R2,
R3 , and Y.
The pharmaceutically acceptable salts of the
compounds of Formula I include the conventional non-
toxic salts or the quaternary ammonium salts of the
compoilnds of Formula I formed, for example, from non-
toxic inorganic or organic acids. For example, such
conventional non-toxic salts include those derived from
inorganic acids such as hydrochloric, hydrobromic,
sulfuric, sulfamic, phosphoric, nitric and the like; and
the salts prepared from organic acids such as acetic,
propionic, succinic, glycolic, stearic, lactic, malic,
tartaric, citric, ascorbic, pamoic, maleic,
hydroxymaleic, phenylacetic, glutamic, benzoic,
salicylic, sulfanilic, 2-acetoxybenzoic, fumaric,
toluenesulfonic, methanesulfonic, ethane disulfonic,
oxalic, isethionic, and the like.
The pharmaceutically acceptable salts of the
present invention can be synthesized from the compounds
of Formula I which contain a basic or acidic moiety by
conventional chemical methods. Generally, the salts are
prepared by reacting the free base or acid with
stoichiometric amounts or with an excess of the desired
salt-forming inorganic or organic acid or base in a
suitable solvent or various combinations of solvents.
The pharmaceutically acceptable salts of the
acids of Formula I with an appropriate amount of a base,
such as an alkali or alkaline earth metal hydroxide e.g.
sodium, potassium, lithium, calcium, or magnesium, or an
organic base such as an amine, e.g.,
dibenzylethylenediamine, trimethylamine, piperidine,
pyrrolidine, benzylamine and the like, or a quaternary
ammonium hydroxide such as tetramethylammoinum hydroxide
and the like.
As discussed above, pharmaceutically-acceptable
= salts of the compounds of the invention can be prepared
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by reacting the free acid or base forms of these
compounds with a stoichiometric amount of the
appropriate base or acid, respectively, in water or in
an organic solvent, or in a mixture of the two;
generally, nonaqueous media like ether, ethyl acetate,
ethannl, isopropanol, or acetonitrile are preferred.
Lists of suitable salts are found in Remington's
Pharmaceutical Sciences, 17th ed., Mack Publishing
Company, Easton, PA, 1985, p. 1418, the disclosure of
which is hereby incorporated by reference.
The disclosures of all of the references cited
herein are hereby incorporated herein by reference in
their entirety.
Synthesis
The compounds of the present invention can be
prepared in a number of ways well known to one skilled
in the art of organic synthesis. The compounds of the
present invention can be synthesized using the methods
described below, together with synthetic methods known
in the art of synthetic organic chemistry, or variations
thereon as appreciated by those skilled in the art.
Preferred methods include, but are not limited to, those
described below. All references cited herein are hereby
incorporated in their entirety herein by reference.
Compounds of Formula I wherein the central
heterocycle is a 3,5-disubstituted isoxazoline ring can
be conveniently prepared by dipolar cycloaddition of
nitrile oxides with appropriate dipolarophiles (for
reviews of 1,3-dipolar cycloaddition chemistry, see 1,3-
Dipolar Cycloaddition Chemistry (Padwa, ed.), Wiley, New
York, 1984; Kanemasa and Tsuge, Heterocycles 1990, 30,
719). The requisite nitrile oxides are in turn prepared 35 from the
corresponding aldehydes via the'intermediate
oximes.
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Scheme I illustrates one synthetic sequence which
will provide the 3,5-isoxazolines of this invention. An
appropriately substituted hydroxylamine is treated with
NCS in DMF according to the method of Liu, et al. (J.
Org. Chem. 1980, 45, 3916). The resulting hydroximinoyl
chloride is then dehydrohalogenated in situ using TEA to
give a nitrile-oxide, which undergoes a 1,3-dipolar
cycloaddition to a suitably`substituted alkene to afford
the isoxazoline. Alternatively, the oxime may be
oxidatively chlorinated, dehydrochlorinated and the
resulting nitrile oxide trapped by a suitable alkene
under phase transfer conditions according to the method
of Lee (Synthesis 1982, 508).
Subsequent hydrolysis of the ester using
conventional methods known to one skilled in the art of
organic synthesis gives the desired acids. Coupling of
the resulting acids to appropriately substituted a- or ~-
amino esters affords an intermediate which can be
deprotected to give compounds of Formula I. The coupling
is carried out using any of the many methods for the
formation of amide bonds known to one skilled in the art
of organic synthesis. These methods include but are not
limited to conversion of the acid to the corresponding
acid chloride, or use of standard coupling procedures
such as the azide method, mixed carbonic acid anhydride
(isobutyl chloroformate) method, carbodiimide
(dicyclohexylcarbodiimide, diisopropylcarbodiimide, or
water-soluble carbodiimides) method, active ester (p-
nitrophenyl ester, N-hydroxysuccinic imido ester)
method, carbonyldiimidazole method, phosphorus reagents
such as BOP-Cl. Some of these methods (especially the
carbodiimide) can be enhanced by the addition of 1-
hydroxybenzotriazole.
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Scheme I
0
R'+_ ~H 1) H EtOH/pyrCl NOH
/n
2) NCS, DMF R'
R1e
R~'r~'~~CO2R Ru R0e
1) TEA,
benzene R'-U-(CH2)n- n COOH
2) UOH, THF, H20 N
R
H2N C02Et RK R16 Ra
1)
[SCC, HOBt, Rg n COO H 2) UOH, -rHF (ey)
N=-O O RS
Alternately, as depicted in Scheme Ia, the above
sequence can be carried out on an aldehyde bearing a
suitably protected functional group which can be
converted into Rl after elaboration of the right hand
side of the target molecules.
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WO 96/37492 PCT/US96/07646
Scheme Ia
0
1) HPOH = HCI NOH
EtOH/pyr
R
R- protected amino group, 2) NCS, DMF CI
i.e., FMOCNH, phthalimido, etc.
R's
R14 'J~C02R
Ru Ris
1) TEA,
PhH R-(CH2),,- q COOH
2) UOH, THF, H20 N-O
R
1) H2N~COdBu Ru Ris R
DCC, HOBt, Rg ~ 'COOtBu
H2N-(CH~,- I
2) deproteatlon N-O 0 R8
N
C~>-- SMe =1-11 Ru R15 R9
1' N N N
c ~>-NH-(CH)2,- COOH
N-O O R8
pyridlne :TFA
2. UGH, aq. THF
3.TFA
Additional isoxazolinyl acetates useful as starting
materials for the preparation of compounds of Formula I,
wherein V is -(phenyl)-Q- and Q is other than a single
bond, can be prepared by cycloaddition of a suitably
substituted chloro or bromooxime with an ester of vinyl
acetic acid as shown in Scheme Ib using literature
methods or modifications thereof. (D. P. Curran & J.
Chao, J. Org. Chem., 1988, 53, 5369-71; J. N. Kim & E.
K. Ryu, Heterocycles, 1990, 31, 1693-97).
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Scheme lb
CO2Me
~OH I PhQ
N
N
C02Me
PhO Br (CI)
Q s S(O)o-2, 0
CO2tBu
N ~-OH I ROC
0 (
ROC Br (CI) NO C02tBu
R=PhorEt
The compounds of the present invention wherein Y is
an oxyalkoxy group, e.g. alkoxycarbonyloxyalkoxy, may be
prepared by reacting a suitably protected carboxylic
acid of Formula I with an e.g. an alkoxycarbonyloxyalkyl
chloride in the presence of an iodide source, such as
tetrabutylammonium iodide or potassium iodide, and an
acid scavenger, such as triethylamine or potassium
carbonate, using procedures known to those skilled in
the art.
The appropriately substituted racemic p-amino acids
may be purchased commercially or, as is shown in Scheme
II, Method 1, prepared from the appropriate aldehyde,
malonic acid and ammonium acetate according to the
procedure of Johnson and Livak (J. Am. Chem. Soc. 1936,
58, 299). Racemic 0-substituted-o-amino esters may be
prepared through the reaction of dialkylcuprates or
alkyllithiums with 4-benzoyloxy-2-azetidinone followed -74-
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by treatment with anhydrous ethanol (Scheme I, Method 2)
or by reductive amination of 0-keto esters as is
described in published PCT patent application W09316038.
(Also see Rico et al., J. Org. Chem. 1993, 58, 7948-51.)
Enantiomerically pure 0-substituted-p-amino acids can be
obtained through the optical resolution of the racemic
mixture or can be prepared using numerous methods,
.
including: Arndt-Eistert homologation of the
corresponding a-amino acids as shown in Scheme II,
Method 3 (see Meier, and Zeller, Angew, Chem. Int. Ed.
Enql. 1975, 14, 32; Rodriguez, et al. Tetrahedron Lett.
1990, 31, 5153; Greenlee, J. Med. Chem. 1985, 28, 434
and references cited within); and through an
enantioselective hydrogenation of a dehydroamino acid as
is shown in Scheme II, Method 4 (see Asymmetric
Synthesis, Vol. 5, (Morrison, ed.) Academic Press, New
York, 1985). A comprehensive treatise on the
preparation of 0-amino acid derivatives may be found in
published PCT patent application WO 9307867, the
disclosure of which is hereby incorporated by reference.
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Scheme II
Method I
R4
HOZCy COH + i) NH,OAC HzN-T-1-CCz
---'~ M e
R4 F{a H 2) MeOH, HCI Ra
t
Method 2
PhyO 1} (R)zCuli H2N\ ^CCz~
0 HN O 2) EtOH, HCI R7a
Method 3
i) IBC_NMM -- O i}Ag. MeOH HzN *
BxHN~CO2H BxHN +CHNz C02M
Ra 2) CH2Nz
R Ra
Method 4
BoCHN` ^COzMe e~~O~~Ve BoCHN`.^COztule
7R/a h~ R7a
The synthesis of N2-substituted diaminopropionic
acid derivatives can be carried out via Hoffman
rearrangement of a wide variety of asparagine
derivatives as described in Synthesis, 266-267, (1981).
The dipolarophiles used to prepare the compounds of
this invention may be prepared by numerous methods. The
w-alkenoic ester class of dipolarophile may be purchased
commercially or prepared by oxidation of the
corresponding w-alkenols by the method of Corey and
Schmidt (Tetrahedron Lett. 1979, 399, Scheme III).
Scheme III
n Cbs = 2 PYr n
~f=J OH _.... ~I~ H
JTO( .
Synthesis of compounds of Formula I which
incorporate the isoxazoline ring in the reverse -76-
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v1'O 96/37492 PCTIUS96/07646
orientation, i.e., a 5,3-disubstituted isoxazoline ring,
is shown in Scheme IV. Cycloaddition of an
appropriately substituted alkene with t-butylformyloxime
Ii
using the method described by Gree et al. (Bioorganic
and Med. Chem. Lett., 1994, 253) provides the
interfnediate t-butyl [5-substituted isoxazolin-3-
yl]acetate. This ester can be converted to compounds of
Formula I using the methods described herein.
Scheme IV
HO\
N
1U~ + ~CO2tBu U
R O
R1/ n N
C02tBu
1. UOH/aq. MeOH
-00- 9
2. R8 RI H R
H2N j CO2Me N CO2H
' Y ~
=HCI R9 U gno,N O Re
TBTU/EtaN
EtOAc or DMF
3. LiOH, aq. THF
Alternately, as illustrated in Scheme IVa, the
reverse isoxazolines may be prepared by reaction of an
appropriate nitro ester with an appropriately
substituted alkene in the presence of a suitable
dehydrating agent such as phenylisocyanate or phosphorus
oxychloride in the presence of an organic amine base,
such as triethylamine or diisopropylethylamine.
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Scheme IVa=
C02Me !;or I/ + FMOCNH ~OCN n N
- dioxane
1. UGH/aq. MeOH
2 R8 H R9
HaN -1-Y 002Me FMOCNH \ q 002Me
-HCI 149 n O N 8
TBTU/Et3N
EtOAc or DMF
H R9
piperidine CN
_NH q (V2H
2 N -TFA n O~N O R8
C )_-e -HI
pyridine
3. UOH, aq. THF
4.TFA
N-protected aminoalkenes useful in the synthesis of
compounds of this invention can be prepared from the
commercially available alcohols as shown in Scheme -~Vb,
via reaction with a suitable activating agent such as p-
toluenesulfonyl chloride in the presence of a base such
as pyridine, followed by displacement with sodium azide
in a suitable solvent such as DMF. Reduction of the
azide by the action of triphenyiphosphine in the presence of water ( for
example see, Scrivern, E. F.V.,
Turnbull, K., Chemical Rev. 1988, 88, 297-360, and the
-78-
SUBSTITUTE SHEET (RULE 26)
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WO 96/37492 PCT1US96/07646
references therein ) provides an amine which is suitably
protected, for example, Fmoc, Boc or phthalimide group
according to literature procedures. (Protecting Groups
in Organic-Synthesis 2nd Ed. Green, T. W., Wits, P. G.
M. pp 309-406. 1991 John Wiley & Sons, Inc. NY)
Scheme IVb
1)TsCI/Pyr
1-3 Q) nJaNJ3
HO \ 3) Ph3P/H2O/EtOH FmocNH
4) Fmoc-CI
The appropriate nitroesters are available from
commercial sources or can be synthesized according to
literature methods (Seebach, D.et al., Chem. Ber. 1982,
115, 1705-1720; Chaser. D.W., Syn. Comm. 1982, 841-842).
Additional methods for the preparation of compounds
of the present invention containing the isoxazoline ring
in the reverse orientaion are outlined in Scheme V. An
appropriate w-alkenol can be reacted with commercially
available ethyl chorooximidoacetate in a suitable
solvent, such as tetrahydrofuran or methylene chloride,
in the presence of a suitable base, such as aqueous
sodium bicarbonate or triethylamine, to provide the
isoxazoline. Alternately the same intermediate is
prepared by heating the alkenol in the presence of
diethylnitromalonate in refluxing mesitylene or decalin,
by the method of Shimizu et al. (Bull. Chem. Soc. Jpn.
1985, 58, 2519-2522.) Oxidation of theresulting alcohol
the corresponding aldehyde can be achieved by nunierous
methods described in the literature (for example see
Comprehensive Organic Transformations by Larock, R.C., p
604, 605, 607-613. VCH publishers, New York, New York,
1989). Reductive amination of the intermediate aldehyde
(for suitable methods see, Abdel-Magid, A. F.,
-79-
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WO 96/37492 PCT/US96/07646
Maryanoff, C. A., and Carson, K.G., Tetrahedron Lett.,
1990, 31, 5595-5598, and references contained therein
with a variety of heteroaryl amines, which may
additionally contain suitable protecting groups,
provides the substituted amines. Alternatively,
deperfding on the nature of the heterocyclic amine, the
reductive amination can be carried out in a two step
procedure, wherein initial formation of an imine is
carried out by treatment of the aldehyde with the
desired amine in the presence of a dehydrating agent
such as magnesium sulfate, sodium sulfate, or molecular
sieves, in a suitable solvent such as carbon
tetrachloride, methylene chloride, benzene or toluene
(for example see, Modern Synthetic Reactions 2nd ed.
House, H. 0., Benjamin/Cummings Publishing Co, Menlo
Park, Ca., 1972.). The imine is then subsequ.ently
reduced with a variety of reducing agents such as sodium
borohydride, sodium cyanoborohydride, or sodium
triacetoxyborohydride, in a suitable solvent such as
methanol, ethanol, tetrahydrofuran, dioxane or 1,2-
dichloroethane, to provide the desired amines.
Hydrolysis of the ester using conventional methods
known to one skilled in the art.of organic synthesis
provides acid intermediates. Coupling of the resulting
acids to the appropriately substituted a- or 0-amino
ester is accomplished using standard coupling reagents,
as described above. The esters may be saponified or in
the case of tert-butyl esters the acid may be produced
either by the action of trifluoroacetic acid with or
without an inert solvent such as met3iylene chloride, or
by the action of anhydrous HC1 in a solvent such as
ether or dioxane. Additional protecting groups may be
removed by methods known to one skilled in the art ( for
example see, Protective Groups in Organic Synthesis 35 2nd,ed. Greene, T. W.,
and Wuts, P. G. M., John Wiley & Sons, Inc. New York, 1991). -80-
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WO 96/37492 PCT/US96/07646
Scheme yc
Ethyl chforooximidoacetate
Aq. NaHCO9 / C02Et
THF Oxidation
or m O,N ~~=-
HO `Im H
: diethyl nitromalonate
mesirylene, 165 C
1. LiOH/aq. M9OH
OH C02Et N CO2Et 2 R8
j~ --a=- F -
m-1 } N ~ m O' N H2N ~C02R
N NH2 Trityl =HCi R9
Trityl BOP-CVNMM
MgS041 CC4 DMF
2. N8CNBH3 R= Me, Et, t-Bu
Rs 0 Rs
N C02R N ~CO2H
N C~N m N H9 ~ ~HR9
N H O Deprotection i i H
Trityt
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WO 96/37492 PCT/US96/07646
A convenient preparation of suitably protected 2-
aminoimidazoles is outlined in Scheme Va.
Scheme Va:
0
. ,
N 1. NaOMe/MeOH N
~ NH2 2. phtha(ic anhydride IC ~-N I
=1/2 H2SO4 mett H
O
O
pyridine
trityl bromide NH2NH2 CN)IIIII DIPEA, CH2C2 N
SEM-Ci O R
R = trityl or SEM R = trityl or SEM
Compounds of formula I wherein b is a double bond
can be prepared using one of the routes depicted in
Scheme VI. Bromination followed by subsequent
dehydrobromination of a suitably substituted methyl 3-
(cyanophenyl)isoxazolin-5-ylacetate, prepared as
described above, using the method of Elkasaby & Salem
(Indian J. Chem., 1980, 19B, 571-575) provides the
corresponding isoxazole intermediate. Alternately, this
intermediate can be obtained by 1,3-dipolar
cycloaddition of a cyanophenylnitrile oxide (prepared
from the corresponding chlorooxime as described in
Scheme I) with an appropriate alkyne to give the
isoxazole directly. Hydrolysis of the ester using
conventional methods known to one skilled in the art of
organic synthesis provides the acid intermediates.
Coupling of the resulting acids to an appropriately
substituted a- or p amino ester is accomplished using
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WO 96/37492 PCT/US96/07646
standard coupling reagents, as described above.
Saponification gives the acids.
Scheme VI.
NOH
y,U ~C02Me
R 'CI -U ~ q COOMe
m Rl
TFA m N IO
1. NBS, AIBN 1) L.iOH
2. KOAc, HOAC 2) DCC,TEA, or TBTU, TEA
reflux R9
H2N
C02M e
Re
Ri -U~ COOMe 3) LiOH
NIO
R9
H
N
Ri'U q COOH
m
N-O O R$
Compounds of Formula I wherein b is a double bond and Q
is oxygen can be prepared from commercially available
methyl 3-hydroxy-5-isoxazolecarboxylate as illustrated
in Scheme VII. Coupling of the hydroxy group to a
suitably N-protected amino alcohol can be achieved in
one step under Mitsunobu reaction conditions (Hughs,
D.L.; Organic Reactions, Volume 42, JoYui Wiley and Sons,
1992, pages 335-656). Alternately, a two step process of
activation of the N-protected aminoalcohol as a aryl or
alkyl sulfonate ester or by conversion to halide
followed by alkylation of the hydroxyisoxazole gives the
same result. Bases suitable for this reaction include
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alkaline hydrogen carbonates, alkaline carbonates,
cesium carbonate, alkaline hydrides, and alkaline
alcoholates such as sodium ethoxide and potassium t-
.
butoxide. The reaction can be run in a number of
different solvents including lower alkyl and branched
alcohols, ethereal solvents, or halocarbons, but it
proceeds most readily in polar aprotic solvents such as
DMF and DMSO. Saponification of the ester using
standard conditions known to one skilled in the art
provides an acid intermediate which can be converted
using the methods described above into compounds of
Formula I.
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Scheme VII
9ocHN. (CH2)n
HO ph3p, DEAD O
C02Me _~ LiOH
N
'O BoCHN-(C))f-CO2Me OHBOCHN, (Cf-12)n R
BoCHN, s
(CH2)n
0 BOP-CIv. 0 H_>--C02Me
N , \ C02H H2N ~C02Me
O R9 N'O O
H2N, (CH2)n R9 I LiOH
H-C02Me
O N~
i BocHN, Rs
N'O 0 (CH2)n H_,,>- CO2H
O N
\
~ H-= N N'O 0
eS N~
~ TFA
H H
N jY N '(CH2)n R9
N 0 i\ ~H ~- C02Me H2N `(CH2)n H R9
C02H
--~ N-
N. 0
O 0 I \
N.
UOH H H Ht =
R9 ~
N j~ N'(CH2)n H C02H MeS -< N
N
N O N pyridine
N'O 0
Additional aldehydes useful for the preparation of
compounds of formula I in the methods depicted in
schemes I-VII can be prepared as illustrated in Scheme
, VIII below:
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Scheme VIII
0
A. 1. i O, tawn., -Hso
0 CHO
0
H2N o~~OH N
2. ewern ox.
0
O `
B. 00
1O' 0 CHO
H2N~~OEt O 0~12. HCI, doxrrohizC OEt
0
C. NH2 NHBOC NHBOC
(BOC)20 DIBAI-H Al
S~ N -~ s N S N
pyricfine toluene
.
~ C02Et C02Et -70 CHO
NHBOC
1. Ph3PCHaOCH3
?hl - S ~ N
2. HaO*
CHO
D.
a-~s Iit. prep. 1. Ph3PaCH2COzE1
~
K Fukase et sl., 2~
H2N N CH3 J. Carbo. Chem. TrHN N CHO
1894, 13:71 b-736
DIBAI-H
0- fN)O'&
TrHN N C02Et toluene TrHN CHO
-70"
Heterocyclic alkenes and aldehydes useful as
intermediates for the preparation of compounds of
formula I in the methods depicted in Schemes I-VII can
be prepared as illustrated in Scheme VIIIa and VIIib.
The carboxylic acids prepared in scheme III, can be
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converted to an acid halide, by a variety of methods,
such as treatment with oxalyl chloride or thionyl
chloride either neat or in the presence of a suitable
solvent such as methylene chloride or toluene, as
outlined in the literature ( for example, see
Comprehensive Organic Transformations by Larock, R.C., p
964-965. VCH publishers, New York, New York, 1989).
Treatment of the acid chloride with diazomethane in a
suitable solvent such as ether, or dioxane, to form the
diazoketone, followed by treatment with HEr ( for
example, see Comprehensive Organic Transformations by
Larock, R.C., p 346. VCH publishers, New York, New York,
1989 ), provides versatile intermediates for the
synthesis of many heterocycles. The examples shown in
scheme VIIIa, are for illustration purposes and do not
constitute a limitation on the scope of the invention.
For example, as illustrated in method A, the a-
haloketone can be treated with r1-acetylguanidine in a
suitable solvent such as acetonitrile from room
temperature to reflux, or N,N-dimethylformamide from
room temperature to 80oC according to the method of
Little, T. L., and Webber, S. E. ( J. Org. Chem., 1994,
59, 7299-7305.) to provide a 2-amino-4-imidazole
derivative. Alternatively, treatment of the a-haloketone
with thiourea, in the presence of a suitable solvent
such as toluene or acetone, at a temperature from 200C
to the boiling point of the solvent, according to the
method of Patt, W. C., Skeean, R. W., and Steinbaugh, H.
A., ( Synnth. Comm. 1990, 20, 3097-3102 ) , provides the
analogous 2-amino-5-thiazole derivative. The alkenes can
be converted by a hydroboration-oxidation procedure (
for example, see Comprehensive Organic Transformations
by Larock, R. C. , p 497-498. VCH publishers, New York,
New York, 1989 ), to provide the alcohols.The alcohols
can be oxidized to the corresponding aldehydes by
numerous published methods (for example see
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Comprehensive Organic Transformations by Larock, R.C., p
604, 605, 607-613. VCH publishers, New York, New York,
1989).
Scheme VIIIa
Oxalyl chloride CH2N2
o
n=1-3
/ HX x
N~~
n p
O
A. N-aceiylguanidine
~\
~--. N H
fl
ACHN
B. Thiourea n
x N
0 H2N
Z= N,S
1) B2Hg/ H202
Z Z 0
N n 2) oxidation ~--N
H2N H2N
Synthesis of pyridyl-containing alkenes and
aldehydes, proceeds as depicted in Scheme VIIIb.The
amino group of 2-amino-6-methylpyridine is protected by
treatment with 2,6-hexanedione, followed by
deprotonation of the methyl group with lithium
diisopropylamide according to the method of Breukelman,
S. P., Meakins, G. D., and Tirel, M. D. ( J. Chem. Soc. Chem. Comm. 1982, 800-
801.). The lithio intermediate can
be trapped with a variety of electrophiles such as
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formaldehyde, ethylene oxide or 3-
(tertbutyldimethylsilyloxy)-1-bromopropane to provide
the alcohol intermediates. Oxidation of the alcohol to
an aldehyde'as described as described in detail above,
followed by olefination reaction under any of a number
of knOwn conditions, such as, for example, a Wittig
reaction or treatment with various titanium reagents
provides the alkene intermediates (see Comprehensive
Organic Transformations by Larock, R.C., p 173-184. VCH
publishers, New York, New York, 1989).
Scheme VIIIb
1) LDA, -78 C to O C
.~ I \
2} electrophile N
N CH3 3) deprotection ~ OH
-'' 1,3
oAdation olefination
~ N O
~" 1,3
N N~
1,3
Alternately, reductive aniination of the appropriate
aldehyde intermediates with heterocyclic amines as shown
in method A and method B of Scheme VIIic, using methods
described above, yields the desired amine intermediates.
Hydroboration-oxidation of the double bond, followed by
oxidation of the alcohol (vide supra) provides
additional aldehyde irrtermediates useful for the
synthesis of compounds of this invention. For
heterocyclic amines unstable to the hydrogen peroxide
required in the hydroboration oxidation procedure, the
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reductive amination can be carried out on appropriate
esterified w-hydroxy aldehydes, as outlined in method B.
Hydrolysis of the esters and oxidation of the alcohol
provides tYie aldehyde.
Scheme VIIic
A. H
o
Rl- NH2 2.3 R'
N~
2) Sodium triacetoxyborohydride H 2,3
1) B2Hs/ H2O2
A', o
N
2) Oxidation H 2,g H
H ~
B. Z= alkyl
o O O
Z Rl Al- NH2 H A
1-4 ~ N ~O Z
2) Sodium triacetoxyborohydride 1-4
1) NaOH Rl- N ~ O
2) Oxidation _4 H
Compounds of Formula I wherein R1 is 2-
iminopyrrolidinyl, 2-iminopiperidinyl, or 2-
iminoazepinyl can be prepared from the commercially
available imides as outlined in Scheme IX.
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Scheme IX
1. O
H Ph3(CH2)2 ~ ~
O N O 0 H
" O N
2. H2 CHO
- n (
3.H30+ n
1. NH2OH H CO2Me 1. Et30+BF4
O N
2. NCS ( \N~O 2. NH3
3. methyl acrylate n 3. (BOC)20
1. LiOH
C02Me 2, H2NCH2CH(R9)CO2tBu
BOCN
1O 3. TFA
N
n
0 COOH
H 9
HN N R
N~1O
n
Additional compounds of formula I where R9 is
-N(R16)R17 can be prepared from the compounds of Schemes
I, IA, IV, IVa, and IVb, wherein R16 is Cbz
(benzyloxycarbonyl),is shown in Scheme X. Selective
removal of the Cbz group may be accomplished by
hydrogenation using palladium suspended on barium
sulfate as the catalyst in a suitable solvent such as
methanol or ethanol, with or without a co-solvent, by
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the method of Nikam, S. S., Kornberg, B.E., Johnson,
D.R., and Doherty, A, M. ( Tetrahedron Lett. 1995, 36,
197-200). Using this method, the Cbz group can be
removed with minimal to no cleavage of the N-O bond
contained in the isozazoline rings shown in Scheme X
example A, and Scheme X example B.
The resulting amines can be converted to additional
compound of formula I by treatment with a wide variety
of reagents, for exantple, acyl halides, chloroformates,
isocyanates, sulfonyichlorides, chiorosulfonamides, and
sulfonylisocyanates, etc. using standard methods.
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Scheme X
A. NHCbz H2 !Pd / BaSO4
N
Ri-U-(CHg)n- q COOR
O-N O Rg R= Me, Et, t-Bu
-
NH2
H
Rj-U-(CH2)1- q C O R
O-N O R8
NHRis (or NHR1$a)
R16CI or R~8aNC0 N
Rrt-U-(CH2)n- COOR
O-"N 0
R8
B.
NHCbz H2! Pd 1 BaSO4
Ri-U-(CH2),:- COOR
N--O 0 R8 R = Me, Et, t-Bu
R14 R1s H NH2
N
Ri-U-(CH2)n- 4 COOR
N'"a 0 R8
R16Ct or R'8sNCO H NHR16 (or NHR18a)
R'-U-(CH~õ- ~1 COOR
N-=O 0
R8
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The detailed processes for preparing the compounds of
Formula I are illustrated by the following Examples. it
is, however, understood that this invention is not limited
to the specific details of these examples. Melting points
are uncorrected. Proton nuclear magnetic resonance spectra
(1H NDiR) were measured in chloroform-d (CDC13) unless
otherwise specified and the peaks are reported in parts per
million (ppm) downfield from tetramethylsilane (TMS). The
coupling patterns are reported as follows: s, singlet; d,
doublet; t, triplet; q, quartet; qt, quintet; m, multiplet.
Example 2
3-[3-[3-(N-3,4,5,6-Tetrahydropyrimidin-2-
yl)aminopropyl]-(5 R,S)-isoxazoline-5-
yl]methylcarbonylamino-2-benzyloxycarbonylamino ro ionic
acid
A. 3-[3-[3-(N-3,4,5,6-Tetrahydro yrimidin-2-
yl)aminopropyl]-(5 R,S)-isoxazoline-5-
yl]methylcarbonylamino-2-benzyloxvcarbonylaminopropionic
acid: The compound of Ex. 16, Part G (199mg; 0.5mmol),
methyl 3-amino-2-benzyloxycarbonylaminopropionate 147mg
(0.5mmol), and triethylamine (100mg; immol) in 3ml
dimethylformamide was treated with (177mg; 0.55mmol) of
O-(1H-benzotriazol-1-yl)-N,N,N1,Nl-tetramethyluronium
tetrafluoroborate (TBTU) and the mixture was stirred at
room temperature for 18 hours. TLC indicated no
starting material; the volatiles were removed under high
vaccum. The residue was purified by flash
chromatography; 55g silica gel column using 8%
MeOH:CHC13 followed by 20% to afford the desired product
as a white solid ( 252mg; 79.9% yield). NMR indicated
contamination with triethylamine salt. The compound
will be used in the next step without further
purification. HRMS calcd. for C24H34N606 ([M+H]+):
503.261808; found: 503.259832.
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B. 3-[3-[3-(N-3,4,5,6-Tetrahydropyrimidin-2-
yl)aminopropyl]-(5 R,S)-isoxazoline-5-
yl]methylcarbonylamino-2-benzyloxycarbonylaminopropionic
acid: A mixture of the compound of Ex. 2, Part A
(100mg; 0.159mmo1) and lithium hydroxide (12mg; 0.5mmol)
in 2ml of a 1:1 methanol:water was stirred at room
temperature for one hour. tLC indicated dissappearance
of starting material. The mixture was diluted with
water and washed with hexane. The aqueous layer was
neutralized with 1N HC1 (0.5mmol) and then stripped.
The residue was purified on a LH2O size exclusion column
using 100% methanol as an eluent. The product obtained
was lyophilyzed from 2ml 1N HC1 followed by 2ml
distilled water. The desired product was obtained as an
off-white solid ( 59mg; 70.7% yield). 1H NMR (300MHz
CDC13): 1.721-1.785 (m, 4H); 2.294-2.408 (m, 3H); 2.499
(dd, 1H, J1=14.28Hz, J2=6.59Hz); 2.675 (dd, 1H,
J1=17.2Hz, J2=6.23Hz); 2.984 (dd, 1H, J1=17.2Hz,
J2=10.25Hz); 3.125 (t, 2H, J=6.59Hz); 3.225 (m, 4H);
3.343-3.558 (m, 2H); 4.214 (m, 1H); 4.768 (m, 1H); 4.973
(s, 2H); 7.145-7.230 (m, 5H). HRMS calcd. for C23H32N606
([M+H]+): 489.246158; found: 489.247644.
Example 16
3- [3-[3-(N-3,4,5,6-Tetrahydropyrimidin-2-
yl ) aminopropyl] -( 5 R, S)- isoxazolin- 5-
yl]methylcarbonylamino-2-phenylsulfonylaminobro ionic
acid
A. N-(4-Hydroxybutyl)phthalimide: A solution of 4-
amino-l-butanol (5.616g; 63mmo1) and triethylamine
(12.3m1; 88.2mmol) in 130m1 tetrahydrofuran was treated
with N-carbethoxyphthalimide (13.82g; 63mmol). The
mixture was refluxed for 18 hours. TLC showed the
formation of product (1:1 Ethyl acetate:Hexane Rf=0.3).
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The mixture was diluted with ethyl acetate and washed
with water. The aqueous layer was extracted with more
ethyl acetate and the organic layers combined, dried
(MgSO4), filtered, concentrated and the residue purified
by flash chromatography; 200g silica gel column using
2:3 ethyl acetate:Hexane followed by 1:1 to provide N-
(4-hydroxybutyl)phthalimide as a white solid (9.108g;
65.9% yield). 1H NMR (300 MHz, CDC13): 1.426 (t, 2H,
J=4.93Hz); 1.597-1.663 (m, 2H); 1.738-1.813 (m, 2H);
3.668-3.768 (m, 4H); 7.715 (m, 2H); 7.838 (m, 2H).
B. N-(4-Oxobutyl)phthalimide: Oxalyl Chloride (4.O1m1;
46mmol) in 90m1 dichloromethane was cooled at -78 C in a
dry ice-acetone bath. Dimethylsulfoxide (4.26m1;
60mmol) in 22m1 dichloromethane was then added dropwise
and the mixture stirred at-78 C for 30 minutes. N-(4-
Hydroxybutyl)phthalimide (9.108g; 41.5 mmol) in 45m1
dichioromethane was then added and the mixture stirred
in the -78 C bath for 45 minutes. The mixture was then
warmed in a 0 C ice bath and stirred for one hour.
Triethylamine (23g; 230mmol) in 23ml dichioromethane was
then added and the stirred for an additional 30 minutes.
The mixture was worked up by washing with water. The
organic layer was separated, dried (MgSO4), filtered,
concentrated and the residue purified by flash
chromatography; 200g silica gel column using 1:3 ethyl
acetate:Hexane to provide N-(4-oxobutyl)phthalimide as a
white solid ( 7.469g; 82.8% yield). 1H NMR (300 MHz,
CDC13): 2.023 (m, 2H); 2.546 (dt, 2H, J1=7.324Hz,
J2=1.098Hz); 3.749 (t, 2H, J=6.958Hz); 7.734 (m, 2H);
7.845 (m, 2H); 9.777 (t, 1H, J=1.099Hz).
C. 4-(N-Phthaloyl)aminobutyraldehyde oxime: N-(4-
oxobutyl)phthalimide (7.46g; 34.3mmol) and triethylamine
(17.2g; 172mmo1) in 75m1 ethanol was treated with
hydroxylamine hydrochloride (11.95g; 172mmol) and
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stirred at room temperature for 2 hours. TLC indicated
no starting material. The solvent was stripped off and
the mixture diluted with ethyl acetate and washed with
water. The organic layer was dried (MgSO4), filtered,
concentrated and the residue dried in vacuo to afford 4-
(N-phthaloyl)aminobutyraldehyde oxime( 7.469g; 73.9%
yield). A 2:=1 mixture of isomers was obtained and the
following is data on the major isomer. 1H NMR (300MHz,
CDC13): 1.856-1.956 (m, 2H); 2.409-2.479 (dt, 2H,
J1=7.691Hz, J2=5.493Hz); 3.713-3.762 (t, 2H, J=7.324Hz);
6.769 (t, 1H, J=5.493Hz) ; 7.720 (m, 2H) ; 7.836 (m, 2H).
D. tert-Butyl 3-[3-(N-Phthaloyl)aminopropyl]-(5R,S)-
isoxazoline-5-yl acetate: N-Chlorosuccinimide (3.39g;
25.4mmol) and 1 drop of pyridine in 50ml chloroform was
treated with 4-(N-phthaloyl)
aminobutyraldehyde oxime (5.89g; 25.4 mmol) in 25m1
chloroform added over a 5 minute period. After the
mixture was completely in solution, it was stirred at
room temperature for 1.5 hours. NMR was used to monitor
the dissappearance of starting material. The mixture
was treated with tert-butyl-3-butenoate (5.42g;
38.1mmo1) followed by triethylamine 2.70g (26.7mmol)
added dropwise over a 2 hour period. The mixture was
stirred at room temperature for 18 hours and worked up
by washing with water, drying the organic layer (MgSO4),
filtering, and concentrating. The residue was purified
by flash chromatography; 200g silica gel using 1:5 Ethyl
acetate:Hexane followed by 1:3 to provide t-Butyl 3-[3-
(N-Phthaloyl)aminopropyl]-(5R,S)-isoxazoline-5-yl
acetate (7.469g; 63.7% yield). 1H NMR (300MHz, CDC13):
1.430 (s, 9H); 1.967 (m, 2H); 2.355-2.476 (m, 3H);
2.624-2.723 (m, 2H); 3.103 (dd, 1H, J1=16.85Hz,
= J2=10.25Hz); 3.730 (t, 2H, J=6.958Hz); 4.820-4.879 (m,
1H) ; 7.712 (m, 2H) ; 7.813 (m, 2H).
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E. tert-Butyl 3-(3-aminopropyl)-(5R,S)-isoxazoline-5-yl
acetate: tert-Butyl 3-[3-(N-phthaloyl)aminopropyl]-
(5R,S)-isoxazoline-5-yl acetate (6.025g; 16.2mmol) in
200m1 ethanol was treated with hydrazine and the mixture
stirred at room temperature for 18 hours. A thick white
prec5:pitate that had formed was filtered off and washed
with more ethanol. The mother liquor was stripped and
diluted with chloroform andwashed with water. The
organic layer was separated, dried (MgSO4), filtered,
concentrated and the residue(3.811g) purified by flash
chromatography; 250g silica gel column using 10%
methanol:chloroform to elute impurities and then 1:1
methanol:chloroform to provide t-Butyl 3-(3-
aminopropyl)-(5R,S)-isoxazoline-5-yl acetate(l.Og; 25.5%
yield). 1H NMR (300MHz, CDC13): 1.398 (bs, 2H); 1.457
(s, 9H); 1.727 (m, 2H); 2.379-2.489 (m, 3H); 2.659-2.784
(m, 4H); 3.114 (dd, 1H, J1=17.21Hz, J2=10.25Hz); 4.816-
4. 919 (m, 1H) . HRMS calcd. for C12H22N203 ([M+H] +) :
243.170868; found: 234.170966.
F. tert-Butyl 3-[3-(N-3,4,5,6-tetrahydropyrimidin-2-
vl)aminopropyl]-(5 R,S)-isoxazoline-5-ylacetate: (tert-
Butyl 3-(3-aminopropyl)-(5R,S)-isoxazoline-5-yl acetate
990mg (4.09mmo1) in 15ml pyridine was treated with 2-
Methylthio-3,4,5,6-tetrahydropyrimidine hydroiodide
1.135g (4.4mmol) and the mixture stirred at reflux for
22 hours. TLC indicated no starting material. The
mixture was stripped and pumped. The residue was
purified by flash chromatography; 130g silica gel column
using 2% MeOH:CHC13 followed by 4% and finally 6% to
elute 923mg of t-Butyl 3-[3-(N-3,4,5,6-
tetrahydropyrimidin-2y1)aminopropyl]-(5 R,S)-
isoxazoline-5-ylacetate(49.9% yield). 1H NMR (300MHz,
CDC13): 1.460 (s, 9H); 1.897-2.001 (m, 4H); 2.435 (t, +
2H, J=6.226Hz); 2.523 (dd, 1H, J1=15.75Hz', J2=6.59Hz); ~
2.662 (dd, 1H, J1=16.11Hz, J2=6.59Hz); 2.779 (dd, 1H,
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J1=17.40Hz, J2=7.69Hz); 3.177 (dd, 1H, J1=17.40Hz,
J2=10.25Hz); 3.299 (dt, 2H); 3.400 (m, 4H); 4.844-4.949
(m, 1H); 7.518 (s, 2H), 7.623 (t, 1H, J=6.23Hz). HRMS
calcd. for 'C16H28N403 ([M+H] +) : 325.223966; found:
325.223355.
v
G. 3-[3-(N-3,4,5,6-tetrahydropyrimi.din-2-
vl)aminoAropyl]-(5 R,S)-isoxazoline-5-ylacetic acid:
~tert-Butyl 3-[3-(N-3,4,5,6-tetrahydropyrimidin-2-
yl)aminopropyl]-(5 R,S)-isoxazoline-5-ylacetate (885mg;
1.96mmo1) was treated with 1:1 dichloromethane:
trifluoroacetic acid (20m1) and stirred at room
temperature for one hour. TLC indicated no starting
material. The mixture was stripped and pumped to afford
3-[3-(N-3,4,5,6-tetrahydropyrimidin-2yl)aminopropyl]-(5
R,S)-isoxazoline-5-ylacetic acid as a brown solid
(777mg; 100% yield). 1H NMR (300MHz DMSO-d6): 1.694 (m,
2H); 1.804 (m, 2H); 2.302 (t, 2H, J=7.32Hz); 2.511 (d,
2H, J=6.59Hz) ; 2.681 (dd, 1H, J1=17.21Hz, J2=7.32Hz) ;
3.037-3.152 (m, 311); 3.231 (m, 4H); 4.693-4.794 (m, 1H);
7.456 (t, 1H, J=5.13Hz) ; 7.788 (s, 2H) . HRMS calcd. for
C12H20N403 ([M+H]+): 269.161366; found: 269.161204.
H. Methyl 3-[3-[3-(N-3,4,5,6-tetrahydropyrimidin-2-
yl)aminopropyl]-(5 R,S)-isoxazoline-5-
yl]methylcarbonylamino-2-phenylsulfonylaminopropionate:
A solution of the compound of Ex. 16, part G (199mg;
0.5mmol), methyl 3-amino-2-phenylsulfonylaminopropionate
(147mg; 0.5mmol), and triethylamine (100mg; immol) in
3m1 dimethylformamide was treated with 265mg (0.6mmol)
of benzotriazol-l-yloxy-tris(dimethylamino)phosphonium
hexafluorophosphate (BOP Reagent) and the mixture
stirred at room temperature for 18 hours. TLC indicated
no starting material; the mixture was pumped to remove
most of the dimethylformamide. The residue was purified
by flash chromatography; 75g silica gel column using 15%
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methanol:chloroform to elute the desired product. NMR
indicated contamination with triethylamine salt and some
form of the BOP Reagent. The compound was then purified
on a LH2O.size exclusion column using 100% methanol as 5 the eluent. 113mg of
desired product was obtained with
slight contamination from triethylamine salts(35.3%
yield). This product was taken on to the next step
without further purification.
I. 3-[3-[3-(N-3,4,5,6-Tetrahydropyrimidin-2-
y1)aminopropyl]-(5 R,S)-isoxazoline-5-
yl]methylcarbonylamino-2-phenylsulfonylaminopropionic
acid: A mixture of the compound of Ex. 16, Part H
(107mg; 0. 168mmol ) and lithium hydroxide (14mg; 0.6mmol)
in 2ml of a 1:1 methanol:water was stirred at room
temperature for one hour. TLC indicated dissappearance
of the starting material. The mixture was diluted with
water and washed with hexane. The aqueous layer was
neutralized with iN HC1 (0.6mmol) and then stripped.
The residue was purified on a LH2O size exclusion column
using 100% methanol as eluent. The product obtained was
lyophilyzed from 2ml iN HC1 followed by 2ml distilled
water. The desired product was obtained as an off-white
solid (68mg; 76.2% yield). 1H NMR (300 MHz CDC13):
1.646-1.797 (m, 4H); 2.244 (m, 3H); 2.322-2.403 (m, 1H);
3.005 (t, 2H, J=6.96Hz); 3.153-3.196 (m, 6H); 3.318 (d,
1H, J=5.86Hz) ; 3.424 (dd, 1H, J1=13.73Hz, J2=4.39Hz) ;
3.835 (m, 1H); 4.693 (m, 1H); 7.303-7.429 (m, 3H); 7.660
(m, 2H) . HRMS calcd. for C21H30N606S ([M+H] +) :
495.202580; found: 495.201869.
Example 56
2-benzyloxycarbonylamino-3-[[3-[4-[(N-imidazolin-2-
yl) amino] butyl] -( 5 R, S)- isoxazolin- 5-yl] carbonylamino] -
propionic acid
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A. 5-Phthalimidopentanol: A mixture of 10.317 g (100
mmol) 5-amino-l-pentanol and 14.812 g (100 mmol)
phthalic anhydride in 200 mL toluene was stirred 18 h
} under nitrogen at reflux while employing a Dean-Stark
trap for removal of water. The reaction was allowed to
cool to room temperature and solvent was removed. The
residue was flash column chromatographed (1;1 hexanes -
ethyl acetate) to provide 19.77 g (84.0 mmol, 84 %) of a
clear liquid; NMR(CDC13): 7.66-7.88 (m, 4H), 3.59-3.75
(m, 4H), 1.32-1.79 (m, 6H); Mass spectrum: m/z 234 (M +
H) .
B. 5-Phthalimidopentanal: A solution of 260 mL
methylene chloride and 10.39 mL (108.22 mmol) oxalyl
chloride was stirred in a 1000 mL round bottom flask
under nitrogen at -78 C. Added over 10 min was 16 mL
dimethyl sulfoxide. Next added over 5 min was 23.61 g
(101.21 mmol) of the product obtained from Ex. 56, Step
A in 60 mL methylene chloride and the mixture stirred
_ 20 for 15 min. Next added was 60 mL (325.0 mmol)
triethylamine and the mixture allowed to warm to room
temperature. The mixture was poured into water,
extracted with three portions of methylene chloride
which were combined, dried, filtered and stripped of
solvent to provide 21.22 g (91.7 mmol, 90%) of a clear
liquid; NMR(CDC13): 9.76 (t, 1H), 7.68-7.90 (m, 4H),
3.73 (t, 2H), 2.50 (t, 2H), 1.60-1.81 (m, 4H); Mass
spectrum: m/z 232 (M + H).
C. 5-phthalimidopentanal oxime: A mixture of 20.60 g
(89.08 mmol) of the product obtained from Ex. 56, Step
B, 250 mL pyridine arid 12.27 g (2 equivs) of
hydroxylamine hydrochloride was stirred 18 hr under
nitrogen at room temperature. Solvent was removed and
the residue triturated under water. The resulting solid
was filtered and suction dried to provide 12.22 g (49.62
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mmol, 55%) of a white solid, mp = 120-123 C;
NMR(CDC13): 7.69-7.90 (m, 4H), 6.70 and 7.40 (two t,
1H), 7.03 (bs, 1H), 1.50-3.77 (m, 8H); Mass spectrum:
m/z 247 (M + H).
D. tert-Butyl 3-[4-phthalimidobutyl3-isoxazolin-5=
(R,S)-yl)-carboxylate: A mixture of 3.50 g (14.212
mmo1) of the product obtained from Ex. 56, Step C, 100
mL N,N-dimethylformamide and 1.897 g ( 14.212 mmol) N-
chlorosuccinimide were stirred for 3 h at room
temperature under nitrogen. Solvent was removed and the
residue flash colunui chromatographed (2:1 hexanes -
ethyl acetate) to provide 3.50 g (12.46 mmol, 87%) of a
clear liquid; NMR(CDC13): 8.50 (bs, 1H), 7.69-7.88(m,
4H),'3.70 (m, 2H), 2.58 (m, 2H), 1.70 (m, 4H) ; Mass
spectrum: m/z 262 ((M + H) - H20). A mixture of 3.50 g
(12.46 mmol) of the product thus obtained, 50 mL
tetrahydrofuran, 25 mL water, 3.0 g (excess) t-butyl
acrylate and 3.0 g (excess) sodium bicarbonate was
stirred 48 h at room temperature under nitrogen. The
mixture was poured into water and extracted with three
portions of ethyl acetate. The combined organic layers
were dried over anhydrous magnesium sulfate, filtered
and solvent removed. The residue was flash column
chromatographed (3:2 hexanes - ethyl acetate) to provide
2.98 g (8.00 mmol, 64%) of a clear liquid; NMR(CDC13) :
7.70-7.86 (m, 4H), 4.83 (m, 1H), 3.72 (t, 2H), 3.15 (m,
2H), 2.42 (t, 2H), 1.60-1.82 (m, 4H), 1.48 (s, 9H); Mass
spectrum: m/z 373 (M + H) .
E. tert-Butyl 3- [4- [ (N-imidazolin-2-yl) amino)butyl] - (5
R,S)-isoxazolin-5-ylcarboxylate, hydroiodide: A mixture
of 2.92 g (7.80 mmol) of the product obtained from Ex. 56, Part D, 100 mL
absolute ethanol and 0.75 mL (3 35 equivs) hydrazine was stirred for 18 h at
room
temperature under nitrogen. Water was added until all
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dissolved. The mixture was extracted with three portions
of ethyl acetate. The combined organic layers were dried
over anhydrous magnesium sulfate, filtered and solvent
was removed. To the residue was added 1.904 g(7.80
mmol) 2-methylthio-4,5-dihydroimidazole hydroiodide and
100 mZ, pyridine. The mixture was set at reflux under
nitrogen for 18 h. The mixture was allowed to cool to
room temperature and the residue flash column
chromatographed (1:4 methanol - chloroform) to provide
0.48 g (1.95 mmol, 25 %) of a gum; NMR(CDC13): 7.90 (bs,
1H), 7.33 (bs, 1H), 4.90 (m, 1H), 1.63-3.80 (m, 14H),
1.47 (s, 9H); Mass spectrum: m/z 311 (free base + H).
F. 3- [4- [(N-imidazolin-2-yl)amino]butyl] - (5 R,S) -
isoxazolin-5-ylcarboxylic acid, trifluoracetate: A
mixture of 480 mg (1.95 mmol) of the product obtained
from Ex. 56, Part E, 15 mL methylene chloride and 1.0 mL
(excess) trifluoroacetic acid was stirred for 18 h at
room temperature under nitrogen. Solvent was removed and
toluene was added. Solvent was removed to provide 240 mg
(0.629 mmol, 32 %) of a gum; NMR(d6-DMSO): 8.27 (m, iH),
7.20 (m, 1H), 4.90 (m, 1H), 1.40-3.70 (m, 14H); Mass
spectrum: m/z 255 (M + H).
G. tert-Butyl 2-benzyloxycarbonylamino-3-[3-[4-[(N-
imidazolin-2-yl)amino]butyl]-(5 R,S)-isoxazolin-5-
yl]carbonylaminopropionate: A mixture of 230 mg (0.603
mmol) of the product obtained from Ex. 56, Part F, 217.8
mg (0.740 mmol) (R)-t-butyl-3-amino-2-
benzyloxycarbonylaminopropionate, 180 mg (0.930 mmol) of
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide and 26 mg
(catalytic) 1-hydroxybenzotriazole hydrate was stirred
at room temperature under nitrogen. Added was 10 mL N,N-
dimethylformamide followed by 170 mg (1.67 mmol)
triethylamine and the mixture stirred 18 h at room
temperature under nitrogen. Solvent was removed and the
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residue flash column chromatographed (1:4
methanol/chloroform) to provide 217 mg (0.329 mmol, 54%)
of a gum; NMR(CDC13/TMS) : 8.36 (bs, 1H) , 7.68 (bs, 1H)
7.58 (bs, 1H), 7.34 (s, 5H), 6.01 (t, 1H), 5.10 (s, 2H),
4.87 (m, 1H), 1.50-4.38 (m, 17H), 1.41 (s, 9H); Mass
spectfum: m/z 531 (free base + H).
H. 2-benzyloxycarbonylamino~-3-[3-[4-[(N-imidazolin-2-
yl) amino] butyl] -( 5 R, S) - isoxazolin- 5-
yl]carbonylaminopropionic acid, trifluoroacetate: A
mixture of 217 mg (0.329 mmol) of the product obtained
from Ex. 56, Step H, 50 mL methylene chloride and 50 mL
of 0.2 M NaOH was placed in a separatory funnel, shaken,
and the layers separated. The organic layer was washed
two more times with 50 mL portions of 0.2 M NaOH. The
organic layer was dried over anhydrous magnesium
sulfate, filtered and solvent was removed. To the
residue was added 10 mL methylene chloride and 0.5 mL
(excess) trifluoroacetic acid and the mixture stirred 18
h at room temperature under nitrogen. Solvent was
removed and toluene was added. Solvent was removed and
the residue triturated under hexanes. The resulting
solid was filtered to dryness to provide 127 mg (0.215
mmol, 65%) of the title compound as an off-white solid,
mp = 100-6 C; NMR(d6-DMSO): 7.21-8.35 (m, 10H), 1.50-
5.12 (m, 20H); Mass spectrum: m/z 475 (M + H).
Example 83
2(S) -Benzyloxycarbonylamino-3- [3- (4- (N- [3, 4, 5, 6-
tetrahydropyrimidin-2-yl]amino)butyl)isoxazolin-5-(R,S)-
ylcarbonyl]aminopropionic acid.
The title compound was prepared in an analogous
manner to the compound of Example 56 by substitution of 2-methylthio-3,4,5,6-
tetrahydropyrimidine hydroiodide 35 for 2-methylthio-4,5-dihydroimidazole
hydroiodide in Ex. 56, Part E. mp 101-108 C.
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Example 110
2 (S) -Benzyloxycarbonylamino-3- [3- (3- (N- [3, 4, 5, 6-
tetrahydropyrimidin-2-yl]amino) ropyl)isoxazolin-5-
(R,S)-ylcarbonyllaminopro ionic acid.
.~ '
A. 4-Phthalimidobutyraldehyde oxime: A solution of 4-
phthalimidobutyraldehyde (R. Hamilton et al.,
Tetrahedron Letters, 1993, 34, 2847) (17.38 g, 80 mmol)
in pyridine (150 mL) was treated with hydroxylamine
hydrochloride (6.67 g, 96 mmol) and stirred at room
temperature for 17 h. After concentration, the residue
was triturated in water, stirred for 3 h, and filtered
to provide the title product as a light tan solid (14.15
g, 76%): NMR (CDC13) S 8.06 (b, 1H), 7.85 (m, 2H), 7.70
(m, 2H), 7.46 (t, 0.15H), 6.76 (t, 0.85H), 3.73 (t, 2H),
2.44 (m, 1.7H), 2.28 (m, 0.3H), 1.90 (m, 2H); mass spec
(NH3-CI) m/z 233 (M+H+, 100%).
B. tert-Butyl 3-(3-[3-Ahthalimidopro yl]-isoxazolin-5-
(R,S)-yl)-carboxylate: A mixture of the product of Ex.
110, Part A (2.10 g, 9.05 mmol), N-chlorosuccinimide
(1.21 g, 9.05 mmol), pyridine (2 drops), t-butyl
acrylate (2.7 mL, 18.10 mmol), and triethylamine (1.5
mL, 10.86 mmol) in 30 mL chloroform was reacted
according to the procedure of Ex. 284, Part D to provide
the title product (2.40 g, 74%): NMR (CDC13) S 7.84 (m,
2H), 7.78 (m, 2H), 4.82 (m, 1H), 3.78 (t, 2H), 3.20 (m,
2H), 2.41 (t, 2H), 2.02 (m, 2H), 1.44 (S, 9H); mass spec
(NH3-CI) m/z 376 (M+Ng4+, 100$).
C. 3- (3- [3-phthalimidopropyl] isoxazolin-5- (R,S) -yl) -
carboxylic acid: The product of Ex. 110 step B (500 mg,
1.40 mmol) was reacted with trifluoroacetic acid (5 mL)
in lOmL methylene chloride according to the procedure of
Ex. 284, Part E to provide 420 mg (100%) of the title
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product as a foamy solid: NMR (DMSO-d6) 8 7.81 (m, 2H),
7.78 (m, 2H), 5.40 (b,1H) , 5.02 (m, 1H), 3.79 (t, 2H),
3.30 (m, 2H), 2.42 (t, 2H), 2.00 (q, 2H).
D. tert-Butyl N?-benzyloxycarbonyl-N3-[3-[3-(3-
phthalimidopropyl) isoxazolin-5- (R, S) -yll carbonyl] -2- (S) -
2,3-diaminopropionate: The product of Ex. 110, Part C
(420 mg, 1.40 mmol) was reacted with t-butyl N2-
benzyloxycarbonyl-2-(S)-2,3-diaminopropionate (412 mg,
1.40 mmol), O-(1H-benzotriazol-1-yl)-N,N,N',N'-
tetramethyluronium tetrafluoroborate(450 mg, 1.40 mmol),
triethylamine (0.6 mL, 4.20 mmol) in 25 mL ethyl acetate
according to the procedure of Ex. 284, Part F to provide
595 mg (66%) of the title product: NMR (CDC13) S 7.81
(m, 2H), 7.70 (m, 2H), 7.34 (s, 5H), 7.04 (b, 1H), 5.06
(s, 2H), 4.90 (m, 1H), 4.38 (m, 1H), 3.70 (m, 4H), 3.20
(m, 3H), 2.39 (bt, 2H), 1.98 (m, 2H), 1.40 (s, 9H); mass
spec (ESI) m/z 579.4 (M+H+, 100%).
E. tert-Butyl N?-benzyloxycarbonyl-N3-[3-[3-(3-
aminopropyl)isoxazolin-5-(R,S)-yl]carbonyl]-2-(S)-2,3-
diaminopropionate: The product of Ex. 110 step D (550
mg, 0.99 mmol) was reacted with hydrazine (0.1 mL, 2.50
mmol) in 5 mL ethanol according to the procedure of Ex.
284 Part G to provide 223 mg (50%) of the title product:
NMR (CDC13) S 7.38 (m, 5H), 7.04 (b, 1H), 5.80 (dd, 1H),
5.10 (s, 2H), 4.90 (m, 1H), 4.38 (m, 1H), 3.64 (m,2H),
3.40-3.12 (m, 2H), 2.76 (m ,2H), 2.40 (m,2H), 1.72 (m,
2H), 1.50 (s, 9H), 1.46 (b, 2H); mass spec (ESI) m/z
449.5 (M+H+, 100%).
F. tert-Butyl N?-benzyloxycarbonyl-N3- [3-(3-(N-
[3,4,5,6-tetrahydropyrimidine-2-yl]amino)propyl)-
isoxazolin-5-(R,S)-ylcarbonyl]-(S)- 2,3-
diaminopropionate: The product of Ex. 110, step E (124 mg, 0.276 mmol) was
reacted with 2-methylthio-3,4,5,6-
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tetrahydropyrimidine hydroiodide (86.0 mg, 0.332 mmol)
in 2 mL pyridine according to the procedure of Ex. 284
Part H to provide 30 mg (25%) of the title product: NMR
(CDC13) b 7.80 (b, 1H), 7.38 (m, 5H), 7.18 (b, 2H),
5.82-5.78 (2b, 1H), 5.10 (s,2H), 4.90 (m, 1H), 4.38 (b,
2H), 3.80 (b, 2H), 3.58-3.10 (m, 9H), 2.42 (b, 2H), 1.95
(b, 2H), 1.42 (s,9H); mass spec (ESI) m/z 531.4 (M+H+,
100$) .
G. N?-benzyloxycarbonyl-N3- [3-(3-(N-[3,4,5,6-
tetrahydropyrimidine-2-yl]amino) ro yl)-isoxazolin-5-
(R,S)-Ylcarbonyl]-(S)- 2,3-diaminopropionic acid: The
product of part F (30mg, 0.051 mmol) was dissolved in
methylene chloride (5 mL) and treated with 0.2 mL
trifluoroacetic acid according to the procedure of Ex.
284, Part I, to provide the title product (25 mg, 90%)
as a glassy foam: NMR (DMSO-d6) b 8.36 (s, 1H), 8.20 (m,
1H), 7.60 (m, 2H), 7.38 (bs, 5H), 7.28 (m, 1H), 5.08 (s,
2H), 4.88 (m, 1H), 4.36 (b, 2H), 3.76 (b, 2H), 3.48-3.08
(9H), 2.30 (b,2H), 1.82 (b, 2H); mass spec (ESI) m/z
475.3 (M+H+, 100$).
Example 284
2(S)-benzyloxycarbonylamino-3-[2-[3-(2-(N-imidazolin-2-
yl)- aminoethyl)isoxazolin-5-(R,S)-
yl]ethylcarbonylaminolpropionic acid.
A. 2-(2-Pthalimidoethyl)-1,3-dioxolane: To a solution
of potassium pthalimide (15.20 g, 82.0 mmol) dissolved
in 150 mL dimethylformamide was added 2-(2-Bromoethyl)-
1,3- dioxolane (14.86 g, 82.0 mmol). After stirring at
room temperature for 22 h, the mixture was diluted with
excess water and stirred. The resulting white
precipitate was collected and dried (17.0 g, 84%): NMR
(CDC13) b 7.84 (m, 2H), 7.72 (m, 2H), 4.98 (t, 1H), 3.99
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(m, 2H), 3.86 (m, 4H), 2.10 (m, 2H); mass spec (NH3-CI)
m/z 248.1 (M+H+ , 100%).
B. 3-Phthalimidopropionaldehyde: The product of Ex.
284, part A (17.0 g, 69.0 mmol) was dissolved in dioxane
(150 inL) and treated with 1:1 1N HC1/water (200 mL).
After stirring'at room temperature overnight, the
mixture was heated to reflux for 3 h. The reaction was
concentrated, neutralized with aqueous NaHCO3,extracted
into chloroform and dried (Na2SO4). Concentration of
the solvent provided the title product (14.0 g, 100%):
NMR (CDC13) 8 7.82 (m, 2H), 7.78 (m, 2H), 4.02 (t, 2H),
2.88 (t, 2H).
C. 3-Phthalimidopropionaldehyde oxime: The product of
Ex. 284, part B (14.0 g, 69.0 mmol) was reacted with
hydroxylamine hydrochloride (5.80 g, 83.0 mmol) in
pyridine (200 mL). After stirring overnight, the
pyridine was evaporated and the resultant mixture
diluted with water. The precipitate was collected and
dried providing the title product as a white solid (8.00
g, 53%): NMR ( CDC13 ) 8 7.82 (m, 2H), 7.76 (m, 2H), 6.82
(t, 1H) , 3.90 (m 4H)
D. tert-Butyl 3-(3-(2-phthalimidoethyll-isoxazolin-5-
(R,S)-yl)-propionate: The product of Ex. 284, part C
(2.69 g, 12.35 mmol) was combined with N-
chlorosuccinimide (1.65 g, 12.35 mmol) and pyridine (2
drops ) in chloroform (30 mL). After stirring at room
temperature for 1 h, t-butyl pentenoate (3.86 g, 24.7
mmol) and triethylamine (2.1 mL, 14.82 mmol) were added
and stirring continued at room temperature. After 18 h,
the resulting mixture was concentrated and flash
chromatographed (7:3 hexane/ethyl acetate) to provide ~
2.50 g (54%) of the title product: NMR (CDC13) 8 7.82 (m, 2H), 7.78 (m, 2H),
4.60 (m, 1H), 3.96 (t, 2H), 3.18
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(dd,1H) 2.72 (m, 3H), 2.38 (dt, 2H), 1.84 (q, 2H), 1.42
(s, 9H); mass spec (NH3-CI) m/z 373.3 (M+H+, 100%).
E. 3- (3- [2-Phthalimidoethvl] -isoxazolin-5- (R,S) -vl) -
propionic acid: The product of Ex. 284, part D (500mg,
1.34 inmol) was dissolved in 10 mL of methylene chloride
and 5 mL trifluoroacetic acid. After 4 h the solution
was concentrated to provide the title product as a foamy
solid (420 mg, 100%): NMR (DMSO-d6) b 7.82 (m,2H), 7.76
(m, 2H), 4.62 (m, 1H), 3.96 (t, 2H), 3.20 (m, 1H), 2.78
(m, 3H), 2.56 (m, 2H), 1.96 (q, 2H).
F. tert - Butyl 2 ( S ) -benzvloxvcarbonylamino - 3 - [ 2 - [ 3 - ( 2 -
phthalimidoethyl)isoxazolin-5-(R,S)-
yl]ethylcarbonylamino]-pro ionate: The product of Ex.
284, part E (420 mg, 1.33 mmol) was combined with t-
butyl N2-benzyloxycarbonyl-2-(S)-2,3-diaminopropionate
(M. Mokotoff and L. Logue, J. Med. Chem., 1981, 24, 554)
(390 mg, 1.33 mmol), O-(1H-benzotriazol-l-yl)-N,N,N1,N--
tetramethyluronium tetrafluoroborate(430 mg, 1.33 mmol),
and triethylamine (0.6 mL, 4.00 mmol) in 25 mL of ethyl
acetate. After stirring at room temperature for 20h, the
reaction was concentrated and flash chromatographed
(ethyl acetate) to provide 647 mg (86%) of the title
product: NMR (CDC13) S 7.82 (m, 2H), 7.72 (m, 2H), 7. 36
(bs, 5H), 6.06 (b,1H), 5.80 (b, 1H), 5.08 (bd, 2H), 4.60
(b, 1H), 4.37 (b, 1H), 3.97 (bt, 1H), 3.62 (m, 1H), 3.07
(m, 1H), 2.70 (b, 3H), 2.24 (b, 1H), 1.97 (m, 1H), 1.44
(s, 9H); mass spec (ESI) m/z 593.4 (M+H+, 100%).
G. tert-Butyl 2(S)-benzyloxycarbonylamino-3-[2-[3-(2-
aminoethyl)isoxazolin-5-(R,S)-
y1]ethylcarbonylamino]propionate: The product of Ex.
284, part F (450 mg, 0.76 mmol) was treated with
hydrazine (0.1 mL, 1.90 mmol) in 7 mL of*ethanol and
stirred at room temperature overnight. The mixture was
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concentrated and taken up in water, the pH was adjusted
to 11 and the resultant extracted with methylene
chloride. The organic layer was dried (Na2SO4),
filtered, and concentrated providing 250 mg (71%) of the
title product as a gummy solid: NMR (CDC13) S 7.40
(m, 5I4) , 6.20 (b, 1H), 5.85 (b, 1H) , 5.10 (s, 2H), 4.60 (b, 1H), 4.30 -(b,
1H), 3.60 (bt, 2H), 3.00-2.96 (m, 3H),
2.60 (2dd, 1H), 2.42 (b, 2H), 2.30 (b, 2H), 2.00-1.80
(m, 2H), 1.58 (bs, 2H), 1.42 (s, 9H); mass spec (ESI)
m/z 463.3 (M+H+, 100%).
H. tert-Butyl 2(S)-benzyloxycarbonylamino-3-[2-[3-(2-
(N-imidazolin-2-ylamino)ethyl)isoxazolin-5-(R,S)-
yllethylcarbonylamino]propionate: The product of Ex,
284, part G (132 mg, 0.290 mmol) was reacted with 2-
methylthio-2-imidazoline hydroiodide (84 mg, 0.342 mmol)
in 5 mL pyridine over an oil bath heated at 120 C .
After 18 h the mixture was cooled and concentrated
providing the title product (102 mg, 66%): NMR (CDC13)
8.08 (b, 1H), 7.60 (b, 1H) , 7.39 (bs, 5H) , 7.20 (b, 1H) ,
6.18 (b, 1H), 5.82 (b 1H), 5.10 (s, 2H), 4.62 (b, 1H),
4.30 (b, IH), 3.61 (bs, 2H), 3.58 (m, 2H), 3.02-2.94 (m,
3H), 2.60 (m, 1H), 2.40 (b, 2H), 2.30 (b, 2H), 1.94 (m,
2H), 1.40 (s, 9H); mass spec (ESI) m/z 531.5 (M+H+,
100$) .
I. 2 ( S ) - benzyloxycarbonylamino - 3 - [ 2 - [ 3 - ( 2 - ( N-
imidazolin-2-yl)- aminoethyl)isoxazolin-5-(R,S)-
yl]ethylcarbonylamino] ro ionic acid: The product of
Ex. 284, part H (100 mg, 0.188 mmol) was dissolved in 2
mL of methylene chloride and 0.2 mL trifluoroacetic
acid. After 5 h, the solution was concentrated and
triturated with ether to provide 70.0 mg (64%) of the =
title product: NMR (DMSO-d6) S 8.20 (m, 1H), 8.04 (m,
1H), 7.53 (bd, 1H), 7.40 (bs, 5H), 5.04 (s, 2H), 4.42
(m,1H), 4.08 (m, 1H), 3.52-3.20 (m, 10H), 3.02 (m, 2H),
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2.60 (m, 1H), 2.12 (m, 2H), 1.70 (m, 2H); mass spec
(ESI) m/z 475.3 (M+H+, 100%).
General Procedure for synthesis of 1-(9-
fluorenylmethoxycarbonylamino)alkenes:
A. 1-(p-Toluensulfonyloxy)-3-butene: 3-Butene-l-ol
(10.5 g, 0.146 mol) was dissolved in 75 mL of pyridine
and cooled in an ice bath. p-Toluenesulfonyl chloride
(28.5 g, 0.150 mol) was added slowly. The solution was
stirred for 8 h in an ice bath then allowed to stir at
room temperature overnight. The solution was poured into
saturated NaHCO3 and ice. After the ice melted the
mixture was extracted with dichloromethane and the
organic layer evaporated to provide the title compound
(28.6 g, 8696).1H NMR (CDC13): S 2.32-2.44 (m, 2H), 2.45
(s, 3H), 4.05 (t, J =10 Hz, 2H), 5.02-5.12 (m, 2H),
5.60-5.74 (m, 1H), 7.35 (d, J= 8 Hz, 2H), 7.78 (d, J
8 Hz, 2H). mass spectrum m/z 311 (M+NH4, base peak), 294
(M+H).
B. 1-amino-3-butene: The product of Ex. 637, part A
(28.6g, 0.126mo1) was dissolved.in 25 mL of
dimethylformamide. Sodium azide (23.5g, 0.354mo1) was
added in several portions and the reaction mixture
allowed to stir at room temperature overnight. The
reaction mixture was poured into 100 mL of water and 200
mL of diethyl ether and the layers separated. The
organic layer was washed with 100 mL of water and 100 mL
of brine and dried over magnesium sulfate. The crude
azide solution was reacted without futher purification.
Triphenylphosphine (34.0 g, 0.129 mol) was added and the
reaction mixture stirred for 6h at room temperature. 2.3
mL of water was added to the reaction and the solution
was stirred overnight. The diethyl ether layer was
distilled and 4.62g (52%)of the title compound was
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obtained. 1H NMR (CDC13): b 1.60 (br s, 2H), 2.20 (q, J
= 9 Hz, 2H), 2.78 (t, J = 9 Hz, 2H), 5.05-5.15 (m, 2H),
5.70-5.84 (m, 1H).
C. 1-(9-Fluorenylmethoxycarbonylamino)-3-butene: The
product of Ex. 637, part B (5.11 g, 65 mmol) was
dissolved in 50 mL of tetrahydrofuran and 50 mL of 10%
NaHCO3-and cooled in an ice bath. 9-
Fluorenylmethoxycarbonyl chloride (16.8 g, 65 mmol) was
added in several portions, after 4h the ice bath was
removed and the reaction mixture allowed to stir at room
temperature overnight. The reaction mixture was poured
into 200 ml of water and extracted with diethyl ether.
The combined organic layers were evaporated to leave a
white solid which was purified by flash column
chromatography (hexane:ethyl acetate 3:1) to yield 5.4 g
(28%) of the desired product. 1H NMR (CDC13) b 2.22-
2.36 (m, 2H), 3.22-3.34 (m, 2H), 4.24 (t, J = 8 Hz, 1H),
4.40 (d, J= 8 Hz, 1H), 4.60 (br s, 1H), 5.06-5.16 (m,
2H), 5.72-5.84 (m, 1H), 7.26-7.44 (m, 4H), 7.58 (d, J
9 Hz, 2H), 7.78 (d, J = 9 Hz, 2H). Mass spectrum: m/z
311 (M+ NH4, base peak), 294, (M+H), HRMS Calcd 294.1494
observed 294.1505.
Example 583
2(S)-Benzyloxycarbonylamino-3-[5-(4-(N-[imidazolin-2-
yl]amino)butyl)isoxazolin-3- (R,S) -
ylcarbonyllaminopropionic acid.
A. 1-(9-Fluorenylmethoxycarbonylamino)-5-hexene: The
title alkene was prepared in 36% yield according to the
method described in the above general procedure, except
starting with 5-hexene-l-ol. 1H NMR (CDC13) S 1.34-1.58
(m, 4H), 2.02-2.14 (m, 2H), 3.12-3.24 (m, 2H), 4.20 (t,
J= 8 Hz, 1H), 4.40 (d, J= 8 Hz, 2H), 4.72 (s, 1H),
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4.90-5.04 (m, 2H), 5.70-5.84 (m, 1H), 7.26-7.42 (m, 4H),
7.58 (d, J = 9 Hz, 2H), 7.76 (d, J= 9 Hz, 2H).
B. 3-Methoxycarbonyl-5-[(9-
fluorenylmethoxycarbonylamino)butyl]-d?-isoxazoline: 1-
(9-Fliuorenylmethoxycarbonylamino)-5-hexene (2.00 g, 6.22
mmol) and phenylisocyanate (3.70 g, 31.11 mmol) was
dissolved in 40 mL of benzene. Thirty drops of
= diisopropylethylamine was added followed by methyl
nitroacetate (1.48 g, 12.44 mmol) and stirred at room
temperature for 48h. The reaction mixture was filtered
and the filtrate evaporated. The residue was purified by
flash column chromatography (hexane:ethyl acetate 3:1 to
1:1) to yield 1.83 g (70%) of a tan solid. 1H NMR
(CDC13) S 1.32-1.84 (m, 6H), 2.78-2.90 (m, 1H), 3.12-
3.34 (m, 3H), 3.86 (s, 3H), 4.20 (t, J = 7 Hz, IH), 4.40
(d, J = 7 Hz, 2H), 4.72-4.86 (m, 2H), 7.26-7.42 (m, 4H),
7.58 (d, J= 9 Hz, 2H), 7.80 (d, J= 9 Hz, 2H). mass
spectrum in/z 440 (M+NH4), 423 (M+H), 244 (base peak).
C. 3-Carboxy-5-[(9-
fluorenylmethoxycarbonylamino)butyl]-d?-isoxazoline:
The product of Ex. 583, Part B (1.83 g, 4.33 mmol) was
dissolved in 50 mL of tetrahydrofuran and 25 mL of water
and cooled in an ice bath. Lithium hydroxide (174 mg,
4.15 mmol) was dissolved in 2 mL of water and added to
the tetrahydrofuran/water solution. After approximately
10min the reaction mixture was quenched with 10% HC1, to
pH=3. The mixture was extracted with diethyl ether,
dried over magnesium sulfate and evaporated to a syrup.
Trituration with benzene:pentane 3:1 and filtration
afforded a yellow solid which was recrystalized from
benzene chloroform 5:1 to yield 1.18g (67%) of the title
compound as a white powder. 1H NMR (CDC13) S 1.34-1.82
(m, 6H), 2.80-2.98 (m, 1H), 3.10-3.32 (m, 3H), 4.16-4.30
(m, 1H), 4.40-4.52 (m, 2H), 4.80-4.90 (m, 2H), 6.50 (br
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s, 1H), 7.26-7.42 (m, 4H), 7.58 (d, J = 9 Hz, 2H), 7.76
(d, J= 9 Hz, 2H). Mass spectrum m/z 426 (M+NH4) 382
(M+NH4-C02, base peak) HRMS calcd 409.1763 observed
409.1748. 5
D. 3=tert-Butyloxycarbonyl-5-[4-(9-
fluorenylmethoxycarbonylamino)butyl]-d?-isoxazoline
The compound of Ex. 583, Part C (715 mg, 1.75 mmol)
was dissolved in 2 mL of dichloromethane and cooled in
an ice bath. 2 mL of ca. 3.5M solution of N,NI-
diisopropyl-O-t-butyl isourea was added and the reaction
mixture stired for 8h, and the ice bath removed, and
stired overnight at room temperature. The reaction
mixture was cooled in an ijce bath and 2 mL of glacial
acetic acid was added dropwise, during which time
vigorous gas evolution occured. The reaction mixture was
diluted with ice water and cautiously neutralized with
saturated Na2CO3, and extracted three times with ethyl
acetate. The organic layer was dried over MgSO4,
filtered and evaporated. The residue was taken up in 20
mL of 1:1 dichloromethane/ ethyl acetate and filtered.
the filtrate was evaporated and purified by flash column
chromatography, CH2C12:hexane:ethyl acetate, 2:2:1 to
yield 375 mg (46%) of the title compound. 1H NMR (CDC13)
S 1.35-1.78 (m, 15H), 2.80, (dd, J = 16, 8 Hz, 1H),
3.15-3.30 (m, 3H), 4.22 (t, J= 7Hz, 1H), 4.40 (d, J
7 Hz, 2H), 4.70-4.80 (m, 2H), 7.28-7.44 (m, 4H), 7.60
(d, J = 9 Hz, 2H), 7.78 (d, J = 9 Hz, 2H). Mass spectrum
m/z 482, (M + NH4), 465, (M+).
E. 3-tert-Butyloxycarbonyl-5-(4-aminobutyl)-0?-
isoxazoline
The compound of Ex. 583, Part D ( 375 mg, 0.81
mmol) was dissolved in 20 mL of dichioromethane, 0.5 mL
of piperidine was added and the reaction mixture stirred
overnight at room temperature. The solvent was -114-
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evaporated and the residue purified by flash column
chromatography CH2C12: isopropanol 2% to CH2C12:
isopropanol 2%: triethylamine: 0.5% to yield 163 mg
(83%) of the title compound. 1H NMR (CDC13) 51.35-
1.80 (m, 15 H), 2.40 (br s, 2H), 2.70-2.86 (m, 3H), 3.22
(dd, J= 16, 8 Hz, 1H), 4.80 (m, 1H). Mass spectrum m/z
243 (M+ H, base peak).
F. 3-tert-Butyloxycarbonyl-5-[4-(imidazolin-2-
ylamino)butyl]-A-2-isoxazoline hydroiodide
The compound of Ex. 583, Part E (163 mg, 0.67 mmol)
and 2-methylthioimidazoline (180 mg, 0.73 mmol) was
dissolved in pyridine and gently ref luxed overnight. The
solvent was evaporated and the residue purified by
preparatory TLC, chloroform, 20% methanol, to yield
100mg (33%) of the title compound. 1H NMR (CDC13) 61.35-
1.80 (m, 15H), 2.84 (dd, J = 16 Hz, 8 Hz, IH), 3.20-3.32
(m, 3H) , 2.56 (br s, 3H), 3.74 (s, 4H) , 4.82 (m, 1H)
Mass spectrum m/z 311 (M (-HI) + H).
G. tert-Butyl-2-benzyloxycarbonylamino-3-[5-[4-[(N-
imidazolin-2-yl)amino]butyl-5(R,S)-isoxazolin-3-
yl]carbonylamino]propionate: The compound of Ex. 583,
Part F (100 mg, 0.23 mmol) was suspended in 2 mL of
dichloromethane and 2 mL of trifluoroacetic acid was
added. The reaction mixture was stirred for 1 h. at room
temperature and the solvent evaporated to give the brown
oil to which was dissolved in 1 mL of DMF. t-Butyl 3-
amino-2-S-(benzyloxycarbonylamino) propionate ( 67 mg,
0.23 mmol) was added followed by benzotriazol-l-yloxy-
tris(dimethylamino)phosphonium hexafluorophosphate (110
mg, 0.24 mmol) and diiosopropylethyl amine ( 65 mg,
0.50 mmol) and stirred at room temperature overnight.
The solvent was evaporated under reduced pressure and
the residue purified by preparatory TLC, chloroform, 20%
methanol to yield 42mg, (28%) of the title compound. 1H
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NMR (CDC13) 81.35-1.80 (m, 15H), 2.80-3.20 (m, 4H), 3.70
(s, 4H), 4.24 (t, J = 5 Hz, 1H), 4.36 (t, J = 5 Hz, 1H),
4.78 (m, 1H), 5.10 (s, 2H), 7.35 (s, 5H).
H. 2 (S) -benzyloxycarbonylamino-3- [5- (4- (N- [imidazolin-
2-yl] amino) butyl) isoxazolin-3 -(R, S) -
ylcarbonyl]aminopropionic acid: The product from Ex.
583, Part G was suspended in 2 ml of dichloromethane and
1 mL of trifluoroacetic acid was added. the reaction
mixture was stired at room temperature for 1 h. The
solvent was evaporated and the residue triturated with
diethyl ether to provide the title compound. 1H NMR
(CDC13) S 1.32-1.76 (m, 6H), 2.80-2.90 (m, 1H), 3.16-3.80
(m, 6H), 4.40-4.52 (m, 2H), 4.70-4.82 (m, 1H), 5.04-5.16
(m, 1H), 7.24-7.42 (m, 5H). mass spectrum m/z 475.4
Example 637
2(S)-Benzyloxycarbonylamino-3-[5-(3-(N-[imidazolin-2-
yl]amino)propyl)isoxazolin-3-(R,S)-
ylcarbonyl]aminopro ionic acid.
A. 1-(9-Fluorenylmethoxycarbonylamino)-4-pentene: The
title alkene was prepared in 49% yield according to the
methods described in the above general procedure, except
starting with 4-pentene-l-ol. 1H NMR(CDC13) S 1.58-1.70
(m, 2H), 2.02-2.16 (m, 2H), 4.22 (t, J = 8 Hz, 1H), 4.42
(d, 8 Hz, 2H), 4.75 (br s, 1H), 4.94-5.08 (m, 2H), 5.72-
5.84 (m, 1H), 7.26-7.42 (m, 4H), 7.58 (d, J = 9 Hz, 2H),
7.78 (d, J = 9 Hz, 2H). Mass spectrum m/z 325 (M=NH4,
base peak) 308, (M+H). HRMS calcd 308.1650, observed
308.1650.
B. 3-Carboxy-5-[(9-
.
fluorenylmethoxycarbonylamino) robyl]-A-2--isoxazoline:
The title compound was prepared according to Ex. 583,
Part B-C in 58% overall yield starting with 1-(9-
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Fluorenylmethoxy-carbonylamino)-4-pentene and methyl
nitroacetate. 1H NMR (DMSO) S 1.30-1.62 (m, 4H), 2.78
(dd J = 16, 8 Hz, 1H), 2.98 (d, J = 7 Hz, 2H), 3.20 (dd,
J= 16, 8 Hz, 1H) , 4.20 (t, J= 7 Hz, 1H), 4.30 (d, J
7 Hz, 2H), 4.68-4.80 (m, 1H, 7.26-7.44, m, 4H), 7.64 (d,
J= 9Hz, 2H), 7.84 (d, J= 9 Hz, 2H). 13.40 (br s, 1H).
Mass spectrum m/z 395 (M+H), HRMS calcd 395.1606
observed 395.1591.
Example 667
2 - (S) -Benzyloxycarbonylami.no-3 - [5 - (R, S) - (4 - (N- (pyridin-
2-yl)amino)butyl)isoxazolin-3-ylcarbonyllaminopropionic
acid hydrochloride salt
A. 3 - Ethoxycarbon;Tl - 5-[- 4-( hydroxy) butyl7 i soxazoline :
5-Hexene-l-ol (5.Og, 0.05M) was dissolved in 30 mL of
tetrahydrofuran, an aqueous solution of NaHCO3 (29.4g,
0.35M in 20 mL water) was added and the reaction mixture
cooled in an ice bath. Ethyl chlorooximinoacetate ( 11.4
g, 0.075M) was added over 15 min. in several portions.
The reaction mixture was stirred in an ice bath for 6h.
An additional amount of ethyl chlorooximinoacetate (
7.75g, 0.05M ) was added and the reaction mixture was
allowed to stir overnight, during which time the ice
bath melted. The reaction mixture was partitioned
between ethyl acetate aaid water and the organic layer
was separated, dried over magnesium sulfate, filtered
and evaporated. The crude product was purified by flash
column chromatography ( hexane / ethyl acetate 3:1 ) to
yield the title compound as a colorless oil (9.22g,
86%). 1H NMR (CDC13): b 1.3 (t, J = 7 Hz, 3H), 1.40-
1.90 (m, 6H) 2.80-2.94 (m, 1H, 3.20-3.54, m, 1Fi), 3.66
(br s, 2H, 4.34, q, J= 7 Hz, 2H), 4.78-4.90 (m, 1H).
mass spectrum m/z 233, (M+IJH4, base peak), 216, (M+)
Alternatively the title compound can be prepared by the
following procedure.
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5-Hexene-l-o1 ( 5 g, 0.05 M) and diethyl
nitromalonate ( 15.4 g, 0.075 M) was dissolved in
mesitylene ( 50 mL ) and refluxed for 5h. The solvent
was removed in vacuo, and the residue purified by flash 5 column
chromatography on silica gel ( hexane / ethyl
acetate 1:1 ) to provide the title product ( 5.11g,
47.5%
) .
B. 3-Ethoxycarbonyl-5-[-4-oxobutyl]isoxazoline:
Oxalyl chloride ( 7.30g, 0.0575M ) was dissolved in
anhydrous methylene chloride and cooled to -60 C in a
dry-ice/CHC13 bath. Dimethylsulfoxide ( 9.38g, 0.12M
was dissolved in anhydrous methylene chloride and added
dropwise over 30 min. to the solution of oxalyl
chloride, and allowed to stir for an additional 30 min.
The product of Ex.667, part A.( 10.75 g, 0.05M ) was
dissolved in anhydrous methylene chloride (30 mL) and
added to the reaction mixture dropwise over 45 min, and
allowed to stirr for an additional 30 min. Triethylamine
( 25.25g, 0.25M ) was added dropwise over 15 min. The
ice bath was removed and the reaction mixture allowed to
warm to room temperature. The reaction mixture was
diluted with methylene chloride ( 100 mL ) and washed
with water, iN HC1, water, and brine. The organic layer
was separated and dried over magnesium sulfate and
evaporated to yield the title compound as a colorless
oil ( 9.549, 90% ). 1H iaMR (CDC13) : b 1.37 (t, J = 7 Hz,
3H), 1.60, 1.92 (m, 6H), 2.52 (t J = 6 Hz, 2H), 2.80-
2.92 (m, 1H, 3.22-3.36, m, 1H), 4.36 (q, J= 7 Hz, 2H),
4.74-4.88 (m, 1H, 9.58, s, 1H).
C. 3-Ethoxycarbonyl-5- [4- (-I?- (pyridin-2-
yl ) amino ) butyl ] i soxaz ol ine :
The product of Ex.667 part B. ( 9.40g, 0.044M ) was {
dissolved in dichloroethane ( 100 mL ) and cooled in an
ice bath. 2-Aminopyridine ( 4.57g, 0.048M ) was added
followed by the addition of sodium triacetoxyborohydride
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( 14.0g, 0.066M ). The ice bath was removed and the
reaction mixture allowed to stir at room temperature
for 4h. The reaction mixture was cautiously poured into
a saturated solution of sodium bicarbonate ( 200 mL ),
and extracted with ethyl acetate. The organic layer was
dried over anhydrous sodium carbonate, filtered and
evaporated to yield a semisolid. The crude product was
triturated with a mixture of ether and hexane, and the
product collected by filtration ( 8.93g, 70% ) 1H DIMR
(CDC13): S 1.30 (t, J= 7 Hz, 3H), 1.40-1.90 (m, 6H,
2.80-2.92, m, 1H), 3.20-3.34 (m, 2H), 4.34 (q, J= 7 Hz,
2H), 4.70-4.90 (m, 2H), 6.30 (d, J= 9 Hz, IH), 6.48
(t, J = 6 Hz, 1H), 7.42 (t J = 6 Hz, 1H), 8.04 (d, J = 4
Hz, 1H).
D. 3-Ethox_ycarbonyl-5- [4- (I7- (pyridin-2-yl) -IT- (tert -
butyloxycarbonyl)amino)butyl)isoxazoline:
The product of Ex. 667 part C. ( 8.93g, 0.031M ) was
dissolved in methylene chloride. 4-Dimethylaminopyridine
( 374mg, 0.003M ) was added followed by di-tert-butyl
dicarbonate ( 14.73g, 0.067M ). The reaction mixture was
allowed to stir at room temperature overnight. The
mixture was diluted with water and the organic layer
separated, dried over magnesium sulfate, filtered and
evaporated. The crude product was purified by flash
column chromatography (hexane; ethylacetate, 3:1) to
yield the title product ( 9.7g, 81% ). 1H NMR (CDC13): b
1.38 (t, J= 7 Hz, 3H), 1.40-1.90 (m, 15H), 2.76-2.86
(m, 1H), 3.18-3.30 (m, 1H), 3.93 (t, J =7 Hz, 2H), 4.32
(q, J= 7 Hz, 2H), 4.70-4.82 (m, 1H) , 7.01 (t, J= 6 Hz,
1H), 7.54-7.86 (m, 2H), 8.36 (d, J = 4 Hz, IH).
E. 5- [4- (Id- (Pyridin-2-yl) -N- (tert-
Y
butyloxycarbonyl)amino)butyllisoxazolin-3-carboxylic
acid: The product of Ex 667 part D, ( 10.7g, 0.027M
was dissolved in a mixture of 20 mL of tetrahydrofuran
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and 20 n1L of water, and cooled in an ice bath. Lithium
hydroxide ( 1,73g, 0.41Li ) was dissolved in 5 ml of
water and added-to the ester solution. the reaction
mixture was stirred for 45m. TLC of the reaction mixture
( hexane/ ethyl acetate, 3:1 ) indicated that no ester
remained. 1M Citric acid solution (40 mL ) was added and the mixture wa- s
extracted several times with ethyl
acetate (until TLC of the organic layer showed no
product). The combined orgainic layers were dried over
magnesium sulfate, filtered, and evaporated and dried
under high vacuum to yield the title product as a light
yellow semisolid ( 9.9g, 99% ).1H NMR (CDC13): S 1.24-
1.82 (m, 12H),
2.78-2.9 (m, 1H) , 3.20-3.32 (m, 1H), 3.90 (t J 7 Hz,
2H), 4.78-4.90 (m, 1H), 7.10 (t J = 6 Hz, 1H), 7.52 (d,
J= 8 Hz, 1H), 7.70 (t, J= 6 Hz, 1H), 8.42 (t, J = 4
Hz, 1H).
Mass spectrum m/z 364.3 ( M+H, base peak
F. tert-Butyl-2-(S)-benzyloxycarbonylamino-3-[5-(R,S)-
(4-(N-( vridin-2-yl)-N-(tert-
butvloxycarbonvl)amino)butylisoxazolin-3-
ylcarbonyl7aminopropionate:
tert-Butyl N2-benzyloxycarbonyl-2-(S)-2,3-
diaminopropionate (M. Mokotoff and L. Logue, J. Med.
Chem., 1981, 24, 554) ( 2.59 g, 8.806 mmol ), and the
carboxylic acid from Ex. 667 part E. ( 3.20 g, 8.806
mmol ) were dissolved in NIT1I-dimethylformamide ( 20mL
). 2dI-methylmorpholine ( 2.72 g, 26.856 mmol ) and
benzotriazol-i-yloxy-tris(dimethylamino)phosphonium
hexafluorophosphate ( 4.09 g, 9.246 mmol ) were added
and the reaction mixture stirred for 48h at room
temperature under nitrogen. Concentration in vacuo to an
orange oil which was purified by silica gel flash column
chromatography ( hexane // ethyl acetate (2:1), to yield
the title compound as a pale yellow syrup ( 4.56 g, 83%
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) .1H ZIR (CDC13) 1.35-1.77 (m, 6H) , 1.48 (s, 9H) ,
1.50 (s, 9H), 2.74 - 2.83 (d of d, J 17.58, 8.05 Hz,
1H) , 3.14 - 3.24 (d of d of d, J 17.58, 8.05, 1.06Hz,
1H), 3.68 - 3.79 (m, 2H), 3.90 - 3.95 (t, J= 6.96 Hz,
2H), 4.35 - 4.41 (m, IH) , 4.67 - 4.79 (m, 1H), 5.11
(s, 2H)(, 5.66 - 5.69 d, J 6.96 Hz, 1H), 6.94 - 6.97
(t, J = 5.36 Hz, 1H), 6.98 ~ 7.06 (d of d, J= 4.76,
1.84 Hz, 1H), 7.28 - 7.37 (m, 5H), 7.51 - 7.59 (t of
d, J 6.96, 1.84 Hz, 1H), 7.58 - 7.67 (t of d, J =
6.96, 1.83 Hz, 1H), 8.35 - 8.39 (d of d, J = 4.76, 1.83,
1H). Mass spectrum m/z = 640.4 (M + H), 264.2 (base
peak).
G. 2- (S) -Benzyloxycarbon_vlamino-3- [5- (R,S) - (4- (N-
(pyridin-2-_vl)amino)butyl)isoxazolin-3-
ylcarbonyl]antinopropionic acid: The product of Ex. 667
part F 60 mg, 0.094 mmol ) was dissolved in 4N HC1 in
dioxane ( 2 mL ), and stirred at room temperature for 5
h. under nitrogen. The solvent was evaporated in vacuo
and the residue purified by reverse phase (C18) HPLC to
yield the title compot,uid ( 39 mg, 87 % ). 1H TIMR
(CDC13): S 1.35-1.77 (m, 6H), 2.74 - 2.83 (m, 2H), 3.20-
3.40 (m, 3H), 3.55-3.80 (m, 3H), 4.40 (br s, 1H), 4.55
(br s, 1H) , 5.02 (t, J= 8 Hz, 2H) , 6.83 (br t, 1H),
7.02 (cd, 6 Hz, 1H), 7.28 (s, 5H), 7.78 (d, J= 4Hz, 1H),
7.85 (t, J= 5 Hz, 1H). Mass spectrum m;z = 484.3.
(M+H).
Example 669
2-(S)-Phenylsulfonylamino-3-[5-(R,S)-(4-(N-(pyridin-2-
yl)amino)butyl)isoxazolin-3-ylcarbonyl]aminopropionic
acid trifluoroacetate salt
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A. Methyl-2- (S) -phenylsulfonylamino-3- [5- (R,S) - (4- (II-
(pyridin- 2 _yl) -N- ( tert -butyloxycarbonyl) -
amino)butylisoxazolin-3-ylcarbonyl]aminopropionate:
Methyl 3-amino-2-phenylsulfonylaminopropionate
Hartman, G.D., Prugh, J. D., Egbertson, M. S., Duggan,
M. E. -Hoffman, W. PCT Int. Appl WO 9408577 Al 940428 ) { ,
8.82 g, 24.3 mmol ) and the product of Ex 667 part E, {
7.15 g,_24.257 mmol ) was disolved in N'N'-
dimethylformamide ( 100mi ). N' -Methylmorpholine (7.61
g, 75.197 mmol) and Benzotriazol-i-yloxy-
tris(di.methylamino)phosphonium hexafluorophosphate
(12.88 g, 29.108 mmol) were added and the reaction
mixture stirred for 15 h. at room temperature under a
nitrogen atmosphere. The solvent was removed in vacuo to
provide the crude product as an orange syrup which was
purified by silica gel flash chromatography ( hexane /
ethyl acetate, 1:1 ), to yield the title compound as a
light yellow oil ( 7.31 g, 57 $). 1H NMR (CDC13) : S
1.34 - 1.77 (m, 6H), 1.50 (s, 9H), 2.59 - 2.68 (d of d,
J = 22.34, 9.52 Hz, 1H), 2.71 - 2.83 (d of d of d, J =
17.58, 8.42, 2.20 Hz, 1H), 2.88 - 2.96 (d, J = 22.34 Hz,
1H), 2.88 - 3.25 (d of d of d, J = 17.58, 10.62, 3.66
Hz, 1H), 3.59 (s, 3H), 3.62 - 3.69 (m, 2H), 3.93 - 3.98
(t, J = 7.33 Hz, 2H), 4.06 - 4.12 (m, 1H), 4.70 - 4.75
(c, 1H), 5.72 - 5.75 (d of d, J = 7.69, 3.36 Hz, 1H),
7.03 - 7.07 (d of d and c, J = 4.76, 1.84 Hz, 2H), 7.47
- 7.50 (t, J = 8.43 Hz, 3H), 7.52 - 7.65 (t, J = 9.15
Hz, 2H), 7.60 - 7.70, (d of d, J = 3.55, 1.83 Hz, 1H),
7.84 - 7.86 (d of d, J = 6.92, 1.46 Hz, 2H), 8.39 - 8.41
(d, J 4.76 Hz, 1H). Mass spectrum m/z = 604.3 (M + H,
base peak).
B. 2- (S) -Phenylsulfonylamino-3- [5- (R,S) - (4- (N- (pyridin- =
2-yl)I,7-(tert-butyloxycarbonyl)amino)butylisoxazolin-3-
ylcarbonyl] aminopropionic acid:
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Lithium hydroxide (0.71 g, 16.952 m.mol) dissolved in
water (10ml). In a separate flask the product of ex 669
part A, ( 7.31 g, 12.109 mmol ) was dissolved in a
mixture of methyl alcohol ( 200mL ), water ( lOmL ). The
lithium hydroxide solution was added and the reaction
mixture was stirred at room temperature for 72 hrs. The
resulting red solution was concentrated in vacuo and
partioned between ethyl acetate ( 100mL ) and water
50mL ). A mixture of 1M Hydrochloric Acid (17m1) in
Citric Acid (100m1) was added until the pH of the aqeous
layer was ca. 4.The organic layer was separated and the
aqueous layer extracted with ethyl Acetate (2 x 30m1).
The combined orgainc layers were dried over anhydrous
magnesium sulfate, filtered and the solvent removed in
vacuo to provide the title compound as a light yellow
oil (6.50 g, 91%).1H NMR (CDC13) : & 1.34 - 1.77 (m, 6H)
1.50 (s, 9H), 2.59 - 2.68 (d of d, J= 22.34, 9.52 Hz,
IH), 2.71 - 2.83 (d of d of d, J= 17.58, 8.42, 2.20
Hz, 1H), 2.88 - 2.96 (d, J= 22.34 Hz, 1H), 2.88 - 3.25
(d of d of d, J= 17.58, 10.62, 3.66 Hz, 1H), 3.59 (s,
3H), 3.62 - 3.69 (m, 2H), 3.93 - 3.98 (t, J= 7.33 Hz,
2H), 4.06 - 4.12 (c, 1H), 4.70 - 4.75 (c, 1H), 5.80 -
5.86 (d of d, J= 7.69, 3.36 Hz, 1H), 6.12 - 6.15 (d, J
= 8.06 Hz, 1H), 7.03 - 7.07 (d of d and c, J= 4.76,
1.84 Hz, 2H), 7.16 - 7.19 (d of d, J= 6.92, 1.46 Hz,
2H), 7.24 - 7.28 (t, J= 8.43 Hz, 3H), 7.38 - 7.57 (t
and d of d, J= 9.15, 4.02, 2.93 Hz, 2H), 7.64 - 7.70
(c, 1H), 7.83 - 7.89 (t, J= 6.95 Hz, 1H), 8.39 - 8.41
(d, J= 4.76 Hz, 1H). Mass spectrum m/z = 590.2 (M + H,
base peak).
C. 2- (S) -Phenylsulfon_vlamino-3- [5- (R,S) - (4- (N- (p_yridin-
2-yl)amino)but_vl.isoxazolin-3-ylcarbonyl]aminopropionic
it
acid
Trifluoroacetic acid (10m1) was added to the product of
Ex 669 part B,( 6.50 g, 11.023 mmol ) and the reaction
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mixture was stirred for 3 h at room temperature under
nitrogen. The solution was concentrated in vacuo then
co-concentrated with toluene ( 3 x 100 mL ). The
resulting oil was purified by gradient reverse phase 5 HPLC (Water /
Acetonitrile) to yield the title compound
as a white powder ( 4.78 g, 72% yield, 97.7 % purity).
1H D]MR (CD30D):. 1.34 - 1.77 (m, 6H) , 1.50 (s, 9H) , 2.59
- 2.68 (d of d, J = 22.34, 9.52 Hz, IH), 2.71 - 2.83 (d
of d of d, J 17.58, 8.42, 2.20 Hz, 1H), 2.88 - 2.96
(d, J = 22.34 Hz, 1H), 2.88 - 3.25 (d of d of d, J =
17.58, 10.62, 3.66 Hz, 1H), 3.30 - 3.36 (t, J = 7.33 Hz,
2H), 3.62 - 3.69 (m, 2H), 4.06 - 4.12 (c, 1H), 4.70 -
4.75 (c, 1H), 5.80 - 5.86 (d of d, J 7.69, 3.36 Hz,
1H), 6.79 - 6.85 (d, J = 4.76 Hz, 1H), 7.00 - 7.03 (d of
d, J = 9.16, 2.56 Hz, 1H), 7.46 - 7.51 (t, J = 7.69 Hz,
2H), 7.57 - 7.59 (t, J = 7.33 Hz, 1H), 7.76 - 7.87 (d, J
= 6. 96, 4H). Mass spectrum m/z = 490.2 (M + H, base
peak). High resolution mass spectrum m/z 490.176031.
Example 695
[2(5)-Benzvloxycarbonvlamino]-3-[5-[(6-aminopyridin-2-
vl)propyl]isoxazolin-3-vl]carbonvlaminopropionic acid,
trifluoroacetate salt:
A. tert-Butyl-[2(S)-Benzyloxycarbonvlamino]-3-[5-[[6-
(2,5-dimethylpyrrolyl)pYridin-2-vl]nropyl]isoxazolin-3-
yl]carbonylaminopropionate: The product of Ex. 697,
Part D (0.43 g, 1.30 mmol ) was dissolved in N,N-
dimethylformamide ( lOmL ).(S)-t-Butyl-3-amino-2-
benyzloxycarbonylamina-propionate ( 0.42 g, 1.43 mmol ),
triethylamine ( 0.29 g, 2.86 mmol ) and O-(1H-
benzotriazol- 1-yl )-Id, N, N' , N' - tetramethyluronium
tetrafluoroborate ( 0.46 g, 1.43 mmol ), according to
the procedure of Ex 284, Part F, to provide the title
=
product ( 0.33g, 42% ). 1HNMR (CDC13) $ 1.46 (s, 9H),
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1.86 (m, 4H) , 2.12 (s, 6H), 2.87 (m, 3H) , 3.22 (m, 1H)
3.74 (br s, 2H), 4.38 (m, lH), 4.78 (m, 1H), 5.11 (s,
2H), 5.68, (bs, 1H), 5.89 (s, 2H), 6.97 (bs, 1H), 7.04
(d, 1H), 7.14 (d, 1H), 7.35 (m, 5H), 7.73 (t, iH). Mass
spectrum: (ESI) m/z 604(M + H)+.
B. [2 (S) -Benzyloxycarbony:Lamino) -3- [5- [ (6-aminopyridin-
2-yl)propyllisoxazolin-3-yllcarbonylaminopro ionic
acid,trifluoroacetate salt: The compound of Ex. 695,
Part A ( 0.33 g, 0.54 mmol ), hydroxylamine
hydrochloride ( 0.75 g, 10.8 mmol, and triethylamine {
0.55 g, 5.42 mmol ) are dissolved in a 4:1 mixture of
isopropanol / water ( 10.OmL ) and refluxed. After 3.5
h, additional hydroxylamine hydrochloride ( 0.37 g, 5.40
mmol ) and triethylamine ( 0.27 g, 2.71 mmol ) are
added, and the reaction mixture refluxed for an
additional 4 h. The reaction mixture was cooled and
sodium carbonate ( 0.86 g, 8.13 mol ) was added. The
reaction mixture was stirred for 16 h, then filtered,
and the filtrate concentrated. The crude amine was
dissolved in a mixture of methylene chloride ( 4.OmL
and trifluoroacetic acid ( 1.OmL ), and stirred for 16
h. The reaction mixture was concentrated to give a
thick yellow oil, which was partitioned between ethyl
acetate and water. The organic layer was separated and
dried over anhydrous sodium sulfate, filtered and
concentrated to give a yellow oil. The crude product
was purified by preparative reverse phase HPLC using a
gradient, 90:10 to 10:90, water / acetonitrile (0.05%
trifluoroacetic acid) as eluent, gave the desired
product as a trifluoroacetate salt, 15 mg ( 5% ). iHNMR
(CDC13) 6 1.70 (m, 4H) , 2.75 (m, 3H) , 3.20 (m, 1H) ,
3.60 (m, 2H), 4.30 (m, 1H), 4.75 (m, 1H), 5.05 (s, 2H),
6.02 (bs, IH), 6.59 (d, J= 8 Hz, IH), 6.71 (d, J = 8
Hz, 1H), 7.35 (m, 5H), 7.68 (t, J= 8 Hz,= 1H). Mass
spectrum: (ESI) m/z 470(M + H)+.
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Example 697
[2-Phenylsulfonylamino]-3-[5-[(6-aminopyridin-2-
.
yl)nropyllisoxazolin-3-yl)carbonylaminopropionic acid, =
trifluoroacetate salt
A. 2-(2,5-Dimethyl-lH-pyrrol-1-Zrl)-6-methylpyridine:
2-Amino-6-methylpyridine (.56.4g , 0.52 mol ), 2,5-
hexanedione ( 59.3 g, 0.52 mol ) and acetic acid ( 5.0
mL ) were refluxed in toluene ( 150 mL ), under a Dean-
Stark trap for 16 h. The mixture was cooled, an
additional portion of 2,5-hexanedione ( 34.0 g, 0.30
mol) was added and reflux resumed for 7 h. The reaction
mixture was cooled, and the solvent was evaporated and
the residue was disolved in EtOAc and cautiously washed
with saturated P7aHCO3,and then brine. The organic layer
was dried with anhydrous sodium sulfate, filtered and
concentrated. The residue was flash column
chromotragraphy ( silica gel, hexane / ethyl acetate,
85:15 ) to provide 37.59 (39%) of the title product.
1H NMR (CDC13) 8 2.12 (s, 6H), 2.59 (s, 3H), 5.88 (s,
2H), 7.02 (d, J = 8 Hz, 1H), 7.15 (d, J = 8 Hz, 1H).
Mass spectrum: (ESI) m/z 187(M + H) +.
B. 2-(2,5-dimethyl-lH-pyrrol-yl)-6-(4=
pentenyl)pyridine:
The product of Ex 697 part A, ( 5.0 g, 0.027 mol ) was
dissolved in anhydrous tetrahydrofuran ( 45mL ) and
cooled to -78 C. A solution of lithium
diisopropylamide ( 0.032mo1 ) in anhydrous
tetrahydrofuran ( SomL ) was precooled to 0 C, and
added to the reaction mixture. After stirring for 1.5 h
at -78 C, 4-bromo-2-butene ( 3.78 g, 0.028 mol ) was
added. This mixture was stirred for an-additional 0.5
h, and then allowed to warm to ambient temperature.
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Saturated ammonium chloride ( 150mL ) was added, the
tetrahydrofuran was evaporated, and the aqueous
solution extracted with ethyl acetate ( 150mL ). The
combined organic layers were washed with water ( 150mL
and brine ( 100mL ), then dried over sodium sulfate,
filtered and concentrated to yield 5.58g (86%) of the
desired product. iH NMR (CDC13) S 1.87 (m, 2H), 2.10
(t, J= 7.3 Hz, 2H), 2.13 (s, 6H), 2.83 (t, J = 7.3 Hz,
2H), 4.99 (m, 2H), 5.83 (m, 1H), 5.89 (s, 2H), 7.02 (d,
J = 7.7 Hz, 1H), 7.13 (d, J = 7.7 Hz, 1H), 7.70 (t, J
7.7 Hz, 1H). Mass spectrum: ( ESI ) m% z 241(M + H) +.
C. 5 -Ethyl [6 - [ [ ( 2 , 5 -dimethylpyrrol_yl) pyridin-2 -
yl]propyl) isoxazolin-3-yl)carboxylate: The product of
ex 697 part B, ( 2.4 g, 9.98 mmol ) was dissolved in a
2:1 mixture of tetrahydrofuran and water ( 90mL ).
Sodium bicarbonate ( 5.01 g, 59.6 mmol ) and ethyl
chlorooximidoacetate ( 1.5g, 9.98mmol ), according to
the procedure of Ex 667, Part A, to provide the title
product ( 1.Olg, 29% ) . 1H I7MR (CDC13) 5 1.36 (t, 3H)
1.67. (m, 2H), 1.88 (m,2H), 2.12 (s, 6H), 2.85 (m, 3H),
3.25 (m, 1H), 4.33 (q, 2H), 4.84 (m, 1H), 5.89 (s, 2H),
7.04 (d, 1H), 7.14 (d, 2H), 7.73 (t, 1H).
D. [6-[[(2,5-Dimethylpyrrol_yl)pyridin-2-yl]propyl]
isoxazolin-3-yl]carboxylic acid: The product of Ex 697
part C, ( 1.01 g, 2.84 mmol ) was hydrolyzed according
to the procedure of Ex 667, Part E, to provide the title
product ( 0.86g, 92% ). 1H NMR (DMSO) 5 1.61 (m, 2H),
1.76 (m, 2H), 2.04 (s, 6H), 2.79 (m, 3H), 3.22 (m, 1H),
4.77 (m, 1H) , 5.78 (s, 1H) , 7.20 (d, IH), 7.30 (d, 1H) ,
7.88 (t, 1H) .
Mass spectrum: (ESI) ntjz 328 (ti + H) +
E. Methyl- [2 (S) -phenylsulfonylamino] -3- [5- [ [6- (2,5-
dimethylpyrrol_vl) pyidin-2 -yl] propyll isoxazolin-3 -
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vl]carbonylaminopropionate: The product of Ex. 697,
Part D ( 0.43 g, 1.30 mmol ) was dissolved in N,N-
dimethyl formamide ( 10mL ), (S)-methyl-3-amino-2-
phenylsulfonylaminopropionate ( 0.42 g, 1.43 mmol ), 5 triethylamine ( 0.29 g,
2.86 mmol ) and O-(1H-
benzotriazol - i-yl )-I\T, N, 2d' , Pd' - tetramethyluronium
tetrafluoroborate ( 0.46 g, 1.43 mmol ), were added
according to the procedure of Ex 284, Part F, to provide
the title product ( 0.23 g, 31% ).iH NMR (CDC13) 5 1.85
(m, 4H), 2.13 (s, 6H), 2.79 (m, 1H), 2.88 (t, 2H), 3.23
(m, 1H), 3.67 (m, 1H), 4.11 (m, 1H), 4.79 (m, 1H), 5.76
(m, 1H), 5.89 (s, 2H), 7.02 (bs, 1H), 7.04 (d, 1H), 7.15
(d, 1H), 7.53 (m, 3H), 7.73 (t, 1H), 7.84 (d, 2H). Mass
spectrum: (ESI) m/z 568 (Ph + H) +.
F. [ 2 ( S ) - Pherry1 su1 f onylamino ] - 3 - [ 5 - [ ( 6 - aminopyr idin - 2
-
v1) rop_yl]isoxazolin-3-yl]carbonylaminopropionic
acid,trifluoroacetate salt: The product of Ex, 697,
Part E, ( 0.16 g, 0.29 mmol ), hydroxylamine
hydrochloride ( 0.40 g, 5.81 mmol ), and triethylamine {
0.29 g, 2.91 mmol ) were dissolved in a 4:1 mixture of
isopropanol / water ( 10.0 mL ) and refluxed . After
3.5 h, additional hydroxylamine hydrochloride ( 0.37 g,
5.40 mmol ) and triethylamine ( 0.27 g, 2.71 mmol ) were
added, and the reaction mixture refluxed for an
additional 4 h. The reaction mixture was cooled and
sodium carbonate ( 0.46 g, 4.36 mol ) was added, the
mixture was stirred 16 h. The reaction mixture was
filtered and concentrated. The crude amine was taken up
in a 4:1 mixture of MeOH / water ( 10.0 mL ) and
LiOH=H20 ( 0.012 g, 0.29 mmol ) was added. The reaction
mixture was stirred 16 h. The methanol was evaporated,
and the crude carboxylic acid was dissolved in water and
washed with EtOAc. The aqueous phase was adjusted to pH
= 4.5 with IM HC1, and the solution absorbed on a pad of
Cia reverse phase gel. The pad was washed well with H20
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and eluted with CH3CN. The CH3CN eluent was
concentrated to give an oily solid. Purification by
preparative reverse phase HPLC using a gradient 90:10
water / acetonitrile to 10:90, water / acetonitrile
0.05% trifluoroacetic acid ) as eluent, gave the desired
product as a TFA salt, i3mg ( 8% ). 1HNMR (CDC13) b
1.80 (m, 4H), 2.81 (m,3H), 3.21 (m, 1H), 3.60 (m, 2H),
4.15 (br s, 1H), 4.80 (br s, 1H), 6.75 (d, J = 4.5 Hz,
1H), 6.85 (d, J = 8.0 Hz, 1H), 7.56 (m, 4H), 7.82 (m,
3H). Mass spectrum: (ESI) m/z 476 (M + H) +.
Example 849
2 - [ ( S ) - ( ( 2 , 4 , 6 - trimethylphenvl ) sulf onyl ) amino l - 3 - [ 5 -
(R,S)-(4-(N-(pyridin-2-yl)amino)butylisoxazolin-3-
vlcarbonyl7 aminopro2ionic acid
A. 2- (S) -amino-3- [5- (R,S) - (4- (N- (pyridin-2-yl) -N- (tert -
butyloxycarbonyl)amino)butylisoxazolin-3-
ylcarbonyl]aminopropionic acid tert-butyl ester:
To the product of example 667 part F, (1.5 g, 2.408
mmol), in methyl alcohol ( 50 mL ) was added Palladium
on Barium Sulfate unreduced ( 0.300 g, 0.482 mmol ).
The reaction mixture was exposed to hydrogen gas ( 41
p.s.i. ) at room temperature for 15 hours. The reaction
mixture was filtered through a celite pad and
concentration in vacuo to afforded a light yellow syrup
(1.22 g, quantitative yield). 1H NMR (CDC13): b 1.34 -
1.76 (m, 6H), 1.46 (s, 9H), 1.48 (s, 9H), 2.29 - 2.48
(br s, 2H), 2.78 - 2.87 (d of d,J = 17.58, 8.05 Hz, 1H),
3.19 - 3.28 (d of d of d, J = 17.58, 8.05, 1.06 Hz, 1H),
3.59 - 3.66 (m, 2H), 3.78 - 3.81 (m, 1H), 3.86 - 3.91
(t, J= 6.96 Hz, 2H), 4.69 - 4.78 (m, IH), 6.98 - 7.02
(d of d, J= 4.76, 1.84 Hz, 1H), 7 12 - 7.14 (t, J
5.36 Hz, 1H), 7.51 - 7.56 (t of d, J= 6.96, 1.84 Hz,
1H) , 7.58 - 7.64 (t of d, J= 6.96, 1.83 Hz, 1H), 8.36 -
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8.38 (d of d, J = 4.76, 1.83 Hz, 1H). Mass Spectrum m/z
= 506.4 (M + H), 197.7 (base peak).
B. 2- [ ( S ) - ( ( 2 , 4 , 6 - trimethylphenyl ) sulf onyl ) amino ] - 3 - [
5 -
(R,S)-(4-(N-(pyridin-2-yl)amino)butylisoxazolin-3-
vlcar-bonvl]aminopropionic acid -
The product of. example 849 part A, (50 mg, 0.0949 mmol),
and pyridine (202 mg, 2.585 mmol) were dissolved in
dichloromethane ( 5 mL ). 2,4,6-trimethylbenzenesulfonyl
chloride (26 mg, 0.119 mmol) was added and the reaction
mixture was stirred at room temperature under nitrogen
for 8 h. Saturated aqueous sodium bicarbonate ( 5 mL
was added and the organic layer was separated, dried
over anhydrous sodium carbonate and concentrated in
vacuo to a syrup. The product was treated with
trifluoroacetic acid (5 ml) at room temperature and
stirred for 1 hour. The reaction mixture was
concentrated in vacuo to a syrup. Toluene ( 10 mL ) was
added and the mixture concentrated in vacuo again. The
resulting syrup was submitted to gradient HPLC (Water /
Acetonitrile) The fractions containing the product were
concentrated in vacuo and placed on a lyophilization
apparatus overnight, to afford the title compound as a
white powder ( 27 mg, 42% over two steps). 1H NMR
(CD3OD): S 1.49 - 1.79 (m, 6H), 2.58 (s, 9H), 2.74-
2.82 (d of d, J = 17.58, 8.05 Hz, 1H), 3.07 - 3.16 ( d
of d of d, J = 17.58, 8.05, 1.06 Hz, 1H), 3.31 - 3.36
(t, J = 6.96 Hz, 2H), 3.31 - 3.65 (d of d, J = 13.55,
5.12 Hz, 2H), 4.00 - 4.06 (m, 1H), 4.74 - 4.82 (m, 1H),
6.78 - 6.85 (d of d, J= 5.86, 0.73 Hz, 1H), 6.94 (s,
2H), 6.98 - 7.03 (d of d, J = 4.40, 0.05 Hz, 1H), 7 75 -
7.78 (t, J = 5.86 Hz, 1H), 7.81 - 7.86 (t, J= 4.40 Hz,
1H). Mass spectrum m/z = 532.2 (M + H base peak).
Example 951
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3-{5-[4-(imidazol-2-ylamino)butyllisoxazoline-3-
carbonYl}amino-2S-(2,4,6-
trimethvlbenzenesulfonylamino)propionic acid trifluoroacetate
~ salt
~ A. 2-Phthalamidoimidazole:
2-Aminoimidazole sulfate ( 2.64 g, 20 mmol ) was dissolved in
200 mL.of anhydrous methanol and cooled to -78'C. A 25%
solution of sodiunt methoxide in methanol ( 4.57 mL, 20 mmol
was added dropwise. The reaction mixture was allowed to warm
to room temperature and stirred for an additional 3 hours.
The solution was filtered and concentrated on the rotary
evaporator to a brown oil ( 1.6g, 96.4% ). 1H NF4R (DMSO) &
5.0 br.s, 2H, 6.32, s, 2H.
Phthalic anhydride ( 4.14 g, 29.2 mmol ) and 2-aminoimidazole
( 2.32 g, 29.2 mmol ) were heated to 170 C for 15 min. The
crude reaction mixture was purified by flash chromatography (
gradient chloroform:methanol 95:5-80:20 ) to yield 4.66 g
(75%) of a brown solid. 1H IdMR (DMSO) S 7.16 (br.s, 2H),
7.94-8.06 (m, 4H), 12.35 (br s, 1H). Mass spectrum ESI
(M+H)t 214.2
B. 1-Triphenylmethyl-2-phthalamidoimidazole.
The product of Ex. 949, Part A, (4.66g, 21.9 mmol) was
dissolved in 100 ntL of anhydrous pyridine and
triphenylmethylchloride ( 9.15 g, 32.82 mmol ) was added. The
reaction mixture was refluxed for 24 hrs. Pyridine was
removed and the residue was purified by flash column
chromatography ( chloroform:methanol 5-10% ) to yield the
desired product ( 2.74 g, 27.5% yield ). IH NMR (CDC13) S
6.80 (d, J = 1.1 Hz, IH), 7.06 (t, J = 7.3 Hz, 3H), 7.17 (t,
J = 7.7 Hz, 7H), 7.28 (d, 6H, 7.64, s 4H). Mass spectrum I3H3-
DCI (M+H) + 456.
C. 1-Triphenylmethvl-2-aminoimidazole.
.
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The product of Ex. 949, Part B, ( 2.60 g, 5.7 mmol ) and
hydrazine ( 1.83 g, 57 mmol ) were refluxed in 250 mL of
anhydrous ethanol for 1 hr. The reaction mixture was cooled
and the solvent removed in vacuo. the solid residue was
purified by flash column chromatography ( chloroform:methanol
10:1 J to yield 1.8 g (97%) of a yellow solid .1H 14MR (DMSO)
S 6.26 (d, J==1.8 Hz, 1H), 6.51 (d, J = 1.8 Hz, 1H) , 7.13
(d, J = 7Hz, 6H), 7.33-7.44 .(m, 9H). Mass spectrum INJH3-DCI
(M+H), 326.
D. 3-Ethoxycarbonyl-5-[4-(1-triphenylmethvlimidazol-2-
ylamino)butyllisoxazoline:
1-Triphenylmethyl-2-aminoimidazole (1.43 g, 4.4 nmmol), the
product of Ex. 667, Part B, ( 1.034 g, 4.85 mmol ) and
magnesiu,~n sulfate ( 2.33 g, 19.4 mmol ) were stirred in 250
mL of carbon tetrachloride at room temperature. Progress of
the reaction was monitored by NMR. After 90 hrs magnesium
sulfate was filtered off and triacetoxyborohydride (3.74 g,
17.65 mmol) was added. The reaction mixture was stirred for
an additional 48 hrs. Water ( 100 mL ) was added, the
organic layer was separated and the water layer was extracted
with ethyl acetate. The combined organic layers were
concentrated and purified by flash chromatography ( ethyl
acetate / hexane 1:1, then ethyl acetate, then ethyl
acetate:methanol 20:1 ) to yield the product as a colorless
oil ( 1.7g, 74% );.1H ITMR (DMSO) S 0.84-0.92 (m, 2H) , 1.0 (m,
2H), 1.26 (t, J 7.3 Hz, 3H), 1.31-1.48 (m, 2H), 2.70 (dd,
J1 = 17.6 Hz, J2 = 8.4 Hz, 1H), 2.90 (q, J 2.3 Hz, 2H),
3.07 (br. s, 1H) 3.20 (dd, Jl = 11Hz, J2 = 17.6 Hz, 1H),
4.25 (q, J = 7.3 Hz, 2H), 4.6 (m, 1H), 6.26 (d, J = 1.5 Hz,
IH), 7.50 (d, J 1.5 Hz, 1H), 7.09 (m, 6H), 7.33-7.44 (m,
6H). Mass spectrum: ISH3 -DCI (M+H) +, 523.
E. 5-[4-il-triphenylmethvlimidazol-2-
ylamino)butyl)isoxazoline-3-carboxylic acid:
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The product of Ex.949, Part D( 246 mg, 0.47 mmol ) was
dissolved in of tetrahydrofuran ( 2 mL ). A 0.5N solution of
lithium hydroxide (0.94 mL. 0.47 mmol) was added. The
reaction mixture was stirred for 1 h, then aqeous 1N HC1
0.47 mL. 0.47 mmol ) was added and the solvent was
evaporated. The crude product was purified by flash column
chromatography 1 chloroform / methanol 10:1 ) to yield the
product as a white solid (230 mg, 99%). 1H PdMR (DMSO) b 0.84-
0.1.31 (m, 6H), 2.57 (dd, Jl = 17.6 Hz, J2 = 8.4 Hz, IH)110 2.89 (q, J= 6.6
Hz, 1H), 3.07 (dd, Jl = 17.6 Hz, J2 = 10.6
Hz, 1 H), 4.0-4.15 (br.s, IH), 4.36 (m, IH), 6.26 (d, J= 1.8
Hz, 1H), 6.52 (d, J= 1.6 Hz, 1H), 7.10 (d, 6H), 7.34-7.44
(m, 9H). Mass spectrum NH3-DCI (M+H)+ -C02H, 451.
G. Id- ( 2, 4, 6 trimethylphenyl ) sulf onyl - L- asparagine
L- Asparagine ( 20.0 g, 0.15M ) was suspended in a mixture of
tetrahydrofuran ( 130 mL ) and water ( 250 mL ).
Triethylamine ( 49 g, 0.48M ) was added followed by
mesitylenesulfonyl chloride ( 49.7 g, 0.227M ), The reaction
mixture became slightly exothermic and the solids dissolved
over a period of 0.5 h. to yield a yellow solution. The
reaction mixture was stirred for 3 h at room temperature,
then washed with ether, and methylene chloride. The aqueous
layer was separated, and acidified to ca. pH = 1.5 with conc.
HCl, during which time a thick precipitate formed. After
0.5h. the product was filtered,washed with water and dried to
yield a white solid (34 g, 72$). m.p.= 193.5 - 1950C 1H PdMR
(DMSO) b 2.24 (s, 3H), 2.28 (dd, IH), 2.45 (dd, 1H), -2.55 (s,
6H), 3.98 (m, 1H), 6.88 (br s, 1H), 6.99 (s, 2H), 7.32 (br s,
1H), 7.82 (d, 2H), 12.58 ;br s, 1H). Mass spectrum ESI m; z=
315.2, (M+H base peak).
H. 3-Amino-2- (S) -rJ- (2,4,6
trimethylphenyl)sulfonylairinopropionic acid
Sodium hydroxide ( 32 g, 0.80 M ), was dissolved in water
200 mL ) and cooled in an ice bath. Bromine ( 19.2g, 0.12 M
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was added dropwise over 5 min. and the mixture allowed to
stir for 15 min. The product of Ex.949, Part G, ( 31.44 g,
0.10 M), was added in several portions over a period of ca.
1'0 min. during which time the yellow color faded. The
reaction mixture was gently heated on a steam bath during
which-time the internal temperature rose to ca. 85 C. After =
ih, the reaction mixture was allowed to cool to room
temperature then cooled in an ice bath. The reaction mixture
was cautiously acidified to pH= 6 with conc. HC1, during
which time a solid formed and gas was evolved. The solid was
filtered, washed with cold water, and allowed to dry
overnight, to yield the product as a white solid ( 23.9 g,
83%- ) . 1H IITMR (DMSO) 5 2.26 (s, 3H) , 2.59 (s, 6H) , 2.80 (dd,
1H), 2.94 (dd, 1H), 3.07 (dd, 1H), 7.06 (s, 2H). Mass
spectrum ESI m/z 287.2 (M+H, base peak).
I. tert-Butyl-3-Amino-2-(S)-r7-(2,4,6-
trimethvl henyl)sulfonylaminopropionate
The product of Ex.949, Part H, ( 11.45 g, 0.04M ), was placed
in a Parr bottle, and dissolved in dioxane ( 170 mL ), and
conc. sulfuric acid ( 11 mL ) was added. The reaction mixture
was cooled in a dry ice / acetone bath and ca. 185 ml of
isobutylene was added. The bottle was sealed and agitated for
114 h. The bottle was de-pressurized then purged with
nitrogen for a brief time. The reaction mixture was poured
into a rapidly stirred mixture of water ( 225 mL ) containing
sodium hydroxide ( 17 g ) and ether ( 600 mL ) which had been
pre-cooled in an ice bath. The layers were separated. The
aqueous layer was extracted with additional ether. The pH of
the aqueous layer was carefully adjusted with conc. HC1 to pH
= 11 and extracted four times with ether. The organic layers
from the pH 11 adjusted extraction were combined, dried with
anhydrous sodium sulfate, filtered and evaporated to yield
the product as a viscous oil which solidified ( 8.64g, 63% ). 35 1H IJMR
(CDC13) b 1.28 (s, 9H), 2.28 (s, 3H), 2.67 (s, 6H),
2.93 (m, 2H), 3.69 (m, 1H), 6.95 (s, 2H).
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J. tert-Butyl 3-f5-[4-(1-triphenylmethylimidazol-2-
ylamino)butyl7isoxazoline-3-carbonyl}amino-2-(S)-(2,4,6-
trimethylbenzenesulfonylamino)propionate.
The product of Ex. 949, Part E, ( 98 mg, 0.198 mmol ) was
dissolved in N,N- dimethylformamide ( 2 mL ). The product of
Ex 949, Part I, (68 mg, 0.198 mmol), O-(1H-benzotriazol-l-
yl) -N,N,N',Nl-tetramethyluronium tetrafluoroborate ( 76 mg,
0.24 mmol ) and triethylamine ( 69 mL, 0.495 mmol ) were
added and the reaction mixture was stirred at room
temperature for 1.5 hrs. The solvent was evaporated and
residue was purified by flash chromatography ( chloroform /
methanol 10:1 ) to yield the product as a white solid (135
mg, 83%). (I,TMR***) Mass spectrum (M+H) + 819.4.
K. 3 - {5 - [4 - (1-Tripheaaylmethyl imidazol - 2 -
ylamino)butyllisoxazoline-3-carbonyl}amino-2-(S)-(2,4,6-
trimethylbenzenesulfonylamino)propionic acid.
The product of Ex. 949, Part J, ( 130 mg, 0.16 mmol ) was
dissolved in a 1:1 mixture of trifluoroacetic acid and
methylene chloride and stirred at room temperature for 5 hrs.
The solvent was evaporated and crude product was purified on
reverse phase HPLC (C18) to provide the desired product ( 75
mg, 73% ). 1H NMR (acetone) b 0.99-1.48, m, 6H, 2.27, s, 3H,
2.62, s, 6H, 2.67, m, 1H, 3.04-3.21, m, 1H, 3.31,q, J = 6.6
Hz, 3.52-3.74, m, 1H, 4.14, m 1H, 4.60, m, 1H, 5.25, br.s,
IH, 6.70, d, J = 2.6 Hz, 6.85, d, 1H, 6.98, s, 2H, 7.14, d, J
= 2.6 Hz, 7.33, m, 6H, 7.48, m, 9H, 7.68, q, 1H; mass
spectrum, NH3-DCI, 763.3, HRMS calc: 763.329094, found:
763. 327781.
L. 3-{5-f4-(imidazol-2-ylamino)butyl)isoxazoline-3-
carbonyl}amino-2S-(2,4,6-
trimethylbenzenesulfonvlamino)propionic acid trifluoroacetate
salt
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The product of Ex. 949, Part K, ( 20 mg, 27 mol ) was
dissolved in trifluoroacetic acid ( 2 mL ). Water ( 0.01 mL
was added and the reaction mixture was refluxed for 1 h. The
trifluoroacetic acid was evaporated and the crude reaction
product was purified on reverse phase HPLC (C18) to yield the
desired prodcut as a white solid ( 10 mg, 58% ). 1H N'MR b
1.34-1.66, m 6H; 2.24, s, 3H, 2.53, s 6H, 2.67-2.76, m, IH,
3.06-3.14, m 1H, 3.20, m, 2H, 3.30-3.48, m, 2H, 3.91, q, 1H,
4.70, m, 1H, 6.50, br,1H, 6.95, s, 2H, 6.95, s, 2H, 7.94, m,
2H, 8.23, m, 1H, 11.9, s, 2H, 12.5-12.8, br., 1H.;Mass
spectrum, ESI, (M+H)+ 521.4, HRMS; calc. 521.219050, found
521.21823.
Example 99911
2 - [ ( S ) - ( ( 2 , 4 , 6 - trimethylphenyl ) sulfonvl ) amina] - 3 - [ 5 -
(R,S)-(4-(N-(3,4,5,6-tetrahydropyridin-2-
yl)amino)butylisoxazolin-3-ylcarbonyl3aminopropionic
acid
The product of example 849, part B ( 12 mg, 0.019 mM )
was disolved in 50 mL of 1:1 ethyl alcohol and 2-
propanol. 5% Palladium on barium sulfate ( 25 mg ) waS
added and the reactioii hydrogenated at 42 psi, at room
temperature for 15h. The reaction mixture was purged
with nitrogen, filtered and concentrated in vacuo. The
resulting oil was purified by reverse phase HPLC
water/ acetonitrile gradient ) to yield the title
compound as a white powder. ( 9 mg, 75% ) 1H NMR (CD3OD)
S 1.41-1.58 (m, 2H, ), 1. 62 - 1.77 (m, 2H) , 1. 7 9-1 . 86 (m, 2H)
2.26 (s, 3H), 2.57-2.64 (m, 2H), 2.59 (s, 6H), 2.74-3.12
(m, 2H), 3.17-3.23 (m, 21H), 3.26-3.32 (m, 2H), 3.34-3.64
(m, 4H), 4.01-4.06 (m,1H0, 4.70-4.80 (m, 1H), 6.95 (s,
2H), 8.24-8.26 (q, 1H), 8.81-8.89 .(br s, 1H); Mass
spectrum, ESI, (M+H)+ 536.5, base peak.
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Example 1001
3- [ [3- [3- [(N-imidazolin-2-yl)amino]propyloxylisoxazol-5-
yllcarbonylaminolpropionic acid
A. Methyl 3-[3-(tert-butyloxycarbonylamino)pro'pyloxy]-
5-isoxazolecarboxylate: Diethylazodicarboxylate (1.46g,
8.39 mmol) was added dropwise to a mixture of methyl 3-
hydroxy-5-isoxazolecarboxylate (1g, 4.55 mmol),
triphenylphosphine (1.46g, 8.39 mmol), and 3-tert-
butyloxycarbonylamino-i-propanol (1.50g, 8.39 mmol) in
anhydrous tetrahydrofuran (10 mL) at 0 C under nitrogen.
After 1 h remove the ice bath and warm to room
temperature overnight. Dilute with ethyl acetate (75
mL) and wash with water (25 mL), saturated NaHCO3 (25
mL), and brine (25 mL). Dry over MgSO4 then evaporate
the solvent in vacuo. The residue was chromatographed
over silica gel (75g, 25 to 60% ethyl acetate/hex) to
give the title compound (1.95g, 95%) as a waxy solid:
1H NMR (300 MHz, CDC13) 8 6.53 (S, 1H) , 4.74 (bs, 1H) ,
4.36 (t, 2H), 3.95 (s, 3H), 3.28 (q, 2H), 2.00 (m, 2H),
1.44 (s, 9H); MS (CI-NH3) m/e 318 (M+NH4) +; HRMS calc'd
for C13H2ON206 + H: 301.1400, found 301.1403.
B. 3-[3-(tert-butyloxycarbonylamino)propyloxyl-5-
isoxazole carboxylic acid: Sodium Hydroxide (0.52g,
13.0 mmol) in water (10 mL) was added in one portion to
the product of Ex. 1001, Part A(1.95g, 6.49 mmol) in
methanol (20 mL) at room temperature. After 2 h the
methanol was removed in vacuo and the residue was taken
up in water (60 mL). The aqueous solution was washed
with ether (30 mL, discard), then acidified to pH<2 with
10% HCl. Extraction with ethyl acetate (3 x 40 mL) was
followed by washing the combined organic extracts with
- brine (40 mL) and drying over MgSO4. The solvent was
evaporated in vacuo to provide the desired-acid (1.60g,
86%) as a white solid: 1H NMR (300 MHz, CDC13)86.93 (s,
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1H), 4.23 (t, 2H) , 3.05 (q, 2H) , 1.82 (m, 2H), 1.37 (s,
9H) ; MS (CI-NH3) nc/e 304 (M+NH4) +; HRMS calc' d for
C12H18N206 + H: 287.1243, found 287.1259.
f
C. Ethyl 3 - [3 - [3 - ( tert -butyloxycarbonyl
amino)propyloxy]isoxazol-5-ylcarbonylamino]propionate:
Diisopropyethylamine (0.68g, 5.24 mmol) was added
dropwise to the compound of Ex. 1001, Part B(0.5g, 1.75
mmol), 0-(1H-benzotriazol-1-yl)-N,N,N',N'-
tetramethyluronium tetrafluoroborate (0.70g, 1,83 mmol),
and ethyl 0-alanine hydrochloride (0.28g, 1.83 mmol) in
anhydrous dichloromethane (30 mL) at 0 C under nitrogen.
After the addition was completed the mixture was allowed
to warm to room temperature and stirred for 7 h. The
reaction was diluted with dichloromethane (75 mL) then
washed with water (25 mL), 5% HC1 (25 mL), saturated
NaHCO3 (25 mL), and brine (25 mL). After drying over
MgSO4 the solvent was evaporated in vacuo and the
residue chromatographed on silica gel (30g, 40 to 75%
ethyl acetate/hexanes) to provide the title compound
(0.23g, 34%) as a viscous oil: 1H NMR (300 MHz, CDCl3) 8
7.05 (bt, 1H), 6.49 (s, 1H), 4.68 (bs, 1H), 4.32 (t,
2H), 4.18 (q, 2H), 3.70 (q, 2H), 3.27 (q, 2H), 2.62 (t,
2H), 1.99 (m, 2H), 1.44 (s, 9H), 1.28 (t, 3H); MS (CI-
NH3) m/e 403 (M+NH4) +; HRMS calc' d for C17H27N307 + H:
386.1927, found 386.1909.
D. Ethyl 3-[3-(3-aminopropyloxy)isoxazol-5-
ylcarbonylamino]propionate: Trifluoroacetic acid (10
ml) was added in a slow stream to the compound of Ex.
1001, Part C (0.18g 0.47 mmol) in dichloromethane (10
mL) at room temperature. After 45 min the
trifluoroacetic acid was removed in vacuo and the
residue was azotopically dried by evaporation in vacuo 35 with toluene (20
mL), then place on a vacuum pump at 0.2
torr overnight. Upon dilution with chloroform the
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product crystallized out and was isolated as a white
solid (0.14g 75%) : 1H NMR (300 MHz, CDC13) 8 8.18 (bs,
3H), 7.80 (bt, 1H), 6.51 (s, 1H), 4.32 (m, 2H), 4.11 (q,
2H), 3.65 (q, 2H), 3.17 (m, 2H), 2.6 (t, 2H), 2.18 (m,
2H) , 1.23 (t, 3H) ; MS (CI-NH3) m/e 303 (M+NH4) +; HRMS
calc'd- for C12H29N305 + H: 286.1403, found 286.1404.
E. Ethyl 3-[3-[3-(imidazoliri-2-yl
amino)propyloxy]isoxazol-5-ylcarbonylamino]propionate:
the compound of Ex. 1001, part D (0.22 g, 0.77 mmol) and
2-methylmercapto-4,5-dihydroimidazole hydroiodide
(0.38g, 1.54 mmol) were combined in ethanol (20 mL) and
heated to reflux for 2 hours. The solvent was removed
in vacuo and the residue was chromatographed over silica
gel (20g, 9:3:1:0.6, chloroform, methanol, water, acetic
acid, lower layer) to provide the desired product (0.13
g, %) as a clear viscous oil: 1H NMR (300 MHz, D4-MeOH)
56.61 (s, 1H), 4.34 (t, 2H), 4.12 (q, 2H), 3.70 (s, 4H),
3.60 (t, 2H), 3.39 (t, 3H), 2.63 (t, 2H), 2.08 (m, 2H),
1,23 (t, 3H) ; MS (CI-NH3) m/e 354 (M+H) +.
F. 3-[3-[3-(imidazolin-2-ylamino)propyloxy]isoxazol-5-
ylcarbonylaminolpropionic acid: Lithium hydroxide
(0.5M, 1 mL) was added to the compound of Ex. 1001, part
E (0.13g, 0.25 mmol) in dioxane (2 mL) at room
temperature. After 1 h the solution was acidified with
HC1 in dioxane (4M, 2 mL) and the solvent was removed in
vacuo. The residue was chromatographed on silica gel
(lOg, 9:3:1:0.6, chloroform, methanol, water, acetic
acid, lower layer) and the product fractions were
evaporated in vacuo. The product was taken up in
methanol (1 mL) and tetrahydrofuran was added slowly.
The mixture was stirred overnight and the title product
- precipitated out as a white solid (32 mg): M.P. 144-
7 C; 1H NMR (300 MHz, d6-DMSO) 6 8. 81 (bs, 2H) , 6.77 (s,
1H), 6.76 (bs, 1H), 4.24 (m, 2H), 3.60 (q, 2H), 3.32 (m,
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4H) , 3.28 (m, 2H) , 2.12 (m, 2H) , 1.81 (m, 2H) ; MS (CI-
NH3) m/e 326 (M+H) +; HRMS calc' d f or C13H19N505 + H:
326.1464, found 326.1462.
F
Example 1003
2(S)--benzyloxycarbonylamino-3-[[3-[2-[(N-imidazolin-2-
yl) amino] ethoxy] isoxazol-5-yl] carbonylamino] propionic
abid
A. [2- (tert-
butyloxycarbonylamino)ethyl]methanesulfonate:
Methanesulfonyl chloride (8.2g 71.6 mmol) in
dichloromethane (45 mL) was added dropwise, over 8 min
at ambient temperature to a stirring solution of 2-(t-
butyloxycarbonylamino)ethan-l-ol and TEA(9.83g, 97.3
mmol) in dichloromethane (150 mL). After 2.5 hr the
reaction mixture was washed with 1N HC1 (2 x 50 mL),
water (2 x 50 mL), and brine (50 mL) then dried over
MgSO4. The solution was filtered, and the solvent
evaporated in vacuo to give the title compound (10.37g,
67%). The resulting waxy solid could be used without
further purification: 1H NMR (300MHZ CDC13) S 4.9(bs,1H),
4.3(t,2H), 3.3(q,2H) 3.04(s,IH).
B. 3-[3-(tert-butyloxycarbonylamino)ethyloxy]-5-
isoxazole carboxylic acid: The compound of Ex. 1003.
Part A(10.37g, 43.3 mmol) in dimethylformamide (25 mL)
was added dropwise over 10 min to a stirring solution of
methyl 3-hydroxy-5-isoxazolecarboxylate and sodium
carbonate (5.83g, 55mmo1). The reaction was heated to
80 C for 3 h, then stirred at ambient temperture for 18
h. 10% Potassium carbonate (- 200ml) was added to the
reaction mixture and was stirred until the resulting
precipitate dissolved. After cooling to 3 C the
solution was acidified to pHN2 with concentrated HC1.
The solid was collected by vacuum filtration, washed =
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with water, then air dried to give the title compound
(5.6g, 47.5%) as a white solid: mp 246 C vigorous
bubbling -160-170 C; 1-H NMR (300MHz, CDC1,3) 8 6.51(s,
1H), 5.02(m, 1H), 4.35(t, 2H), 3.55(m, 2H), 1.43(s, 1H).
C. Methyl 2-benzyloxycarbonylamino-3-[3-[3-(tert-
~
butyloxycarbonylamino)propyloxy]isoxazol-5-
ylcarbonylaminolpropionate: Triethylamine (1.7g,
16.7mmo1) was added in one portion to a mixture of the
compound of Ex. 1003, Part'B (1.68g, 6.2 mmol), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(1.8g, 9.3 mmol), the compound of Ex. 1004, part A
(2.14g, 7.4mmo1) and hydroxybenztriazole (260mg 1.9mmol)
in anhydrous dimethylformamide (10 mL) at ambient
temperature. After 14 h the reaction was diluted with
1N HC1 to six time the volume. The acidic solution was
then extracted with ethyl acetate (5 x 30 mL) and the
combined organic extracts were washed with 10% K2C03(3 x
50 ml), water(2 x 25 mL), and brine (25 mL). After
drying over MgSO4 the solvent was evaporated in vacuo
The crude product was triturated in 10% K2C03, filtered,
washed with water then air dried. The product thus
obtained (1.85g 59%) was isolated as a white crystaline
solid and could be used with out further purification:
1H NMR (300MHz, DMSO ) a 8.95 (m, 1H), 7.78 (d, 1H),
7.36-7.31 (m, 5H) 7.05 (m, 1H), 6.79 (s, IH) 5.09-4.99
(dd, 2H), 4.3,5-4.28 (q, 1H), 4,23-4.19 (t, 2H), 3.62 (s,
3H), 3.58-3.56 (m, 2H), 3.32-3.28 (m, 2H), 1.37 (s,9H);
MS (DCI-NH3) m/e 524 (M+NH4)+.
D. 2-benzyloxycarbonylamino-3-[3-(2-tert-
butyloxycarbonylamino)ethyloxylisoxazol-5-yl carbonyl
amino]propionic acid: The compound of Ex. 1003, Part C
(1.8 g, 3.6 mmol) was saponified using the procedure
outlined for Ex. 1, Part C. The acid (0.81 g, 92%) was
isolated as a brittle foam: 1H NMR (300 MHz, DMSO)58.92
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(m, 1H) 7.62 (d, 1H) 7. 35-7 .30 (m, 5H) 7. 05 (m, 1H) , 6.78
(s, 1H) 5.08-4.97 (dd, 2H) 4.27-4.19 (m, 3H) 3.60-3.55
(m 2H) 3.31-3.28 (m, 2H) 1.37 ( s, 9H); MS (DCI-NH3) m/e
5'10 ( M+NH4 ) + . ~
E. 3--[3-(2-aminoethyloxy])isoxazol-5-ylcarbonylamino]- 2-
benzyloxycarbonylaminopropionic acid hydrochloride: A
solution of HC1 in dioxane (4M, 10 mL) was added to the
compound of Ex. 1003, Part C (1.6 g, 3.25 mmol) in lOm1
of dioxane. The mixture was stirred for 3 h at room
temperature at which time a white solid had precipatated
out. The product was filtered and washed with ether
then dried overnight under vacuum (1.25 g, 90%): 1H NMR
(300 MHz, DMSO)S 12.83 (bs, 1H), 9.06 (t, 1H), 8.24 (bs,
3H), 7.64 (d, 1H), 7.32 (m, 5H), 6.87 (s, 1H), 5.00 (s,
2H), 4.42 (t, 2H), 4.22 (m, 1H), 3.56 (m, 2H), 3.21 (m,
2H) ; MS (CI-NS3) ; HRMS calc'd for C17H21N407 + H:
393.1410, found 391.1391.
F. 2(S)-benzyloxycarbonylamino-3-[3-[2-(imidazolin-2-yl
amino)ethyloxy]isoxazol-5-yl carbonyl amino]propionic
acid: 2-methylthioimidazolinium iodide (114mg,
0.5mmo1), the compound of Ex. 1003, Part E(0.10g,
0.23mmol), and dimethylamino pyridine (60mg) were taken
up in lml of pyridine then heated to reflux for 2-3 min.
The reaction was cooled to between 70-80 C and stirred
for 24hr. The solvent was removed in vacuo and the
residue chromatographed on silica gel (20g, 9:3:1:0.6,
chloroform, methanol, water, and acetic acid, bottom
layer). The product fractions were evaporated in vacuo
and the resulting solid recrystallized from
methanol/acetone to yield the title compound
(33mg,31.2%) as a tan solid: mp 226.8 C dec; 1H NMR
(300MHz, DMSO) S 8. 82 (m, 1H), 7.33 (m, 5H) 6.75 (d, 1H)
4.99 (s, 2H) 4.30-4.27(m, 2H), 3.58-3.52(m,7H),
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3.33(m,1H); Mass Spec (ESI) m/e 461 (M+H)+; HRMS calcld
for C20H25N607 + H: 461.1784, found 461.1789.
Example 1004
2(S)-benzyloxycarbonylamino-3-[[3-[3-[(N-imidazolin-2-
yl)amiho]propyloxy]isoxazol-S-ylJcarbonylaminoJpropionic
acid
A. Methyl 3-amino-2(S)-
(benzyloxycarbonyl)aminopropionate hydrochloride: A
solution of 4N HC1 in dioxane (20 mL) was added to 3-
amino-2-(benzyloxycarbonyl)aminopropanoic acid (2.39g,
10 mmol) in methanol (20 mL) and the solution was
stirred for 2 hours. The solvents was removed in vacuo
to give the methyl ester (2.74 g, 95%) as a white solid
product. NMR (DMSO-d6): b 8.38 (b, 3H); 7.96 (d, 1H);
7.38 (m, 5H); 5.05 (s, 2H); 4.44 (m, 1H); 3.66 (s, 3H);
3.14 (m, 2H)
B. Methyl 2-benzyloxycarbonylamino-3-[3-[3-(tert-
butyloxycarbonylamino)propyloxyJisoxazol-5-
yicarbonylamino7propionate: Diisopropylethylamine (0.54
g, 4.17 mmol) was added dropwise to a mixture of the
compound of Ex. 1001, Part B (1.0 g, 3.48 mmol), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(0.70g, 3.65 mmol), and the compound of Example 1004,
Part A(1.Og, 3.48 mmol) in anhydrous dimethylformamide
(10 mL) at 0 C under nitrogen. After 48 h the reaction
was diluted with water (75 mL) then extraced with ethyl
acetate (3 x 30 mL). The combined organic extracts were
washed with water (2 x 25 mL) and brine (25 mL). After
drying over MgSO4 the sovent was evaporated in vacuo and
the residue chromatographed on silica gel (30g, 30 to
75% ethyl acetate/hexanes) to provide the title compound
(0.90g, 50%) as a viscous oil: 1H NMR (300 MHz, CDC13)6
7.35 (m, 5H), 7.03 (bt, 1H) , 6.49 (s, 1H), 5.78 (d, 1H),
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5.13 (s, 1H), 4.73 (bt, 1H), 4.54 (q, 1H), 4.35 (t, 2H),
3.86 (t, 2H) , 3.79 (s, 3H) , 3.28 (q, 2H) , 1.99 (m, 2H),
1.44 (s, 9H); MS (CI-NH3) m/e 538 (M+NH4)+; HRMS calc'd
for C24H32N409 + H: 521.2248, found 521.2266.
C. 3= [3- [3- (tert-
butyloxycarbonylamino)propyloxy]isoxazol-5-yl carbonyl
amino]-2-benzyloxycarbonylami.nopropionic acid: The
compound of Ex. 1004, Part B(0.9g, 1.73 mmol) was
saponified using the procedure outlined for Example
1001, Part C. The acid (0.81g, 92%) was isolated as a
brittle foam: 1H NMR (300 MHz, DMSO) b 8.91 (bt, 1H),
7.62 (d, 1H), 7.34 (m, 5H), 6.91 (bt, 1H), 6.78 (s, 1H),
5.03 (dd, 2H), 4.22 (m, 3H), 3.59 (m, 2H), 3.05 (q, 2H),
1.84 (m, 2H), 1.37 (s, 9H); MS (CI-NH3) m/e 524
(M+NH4) +; HRMS calc'd for C23H30N409 + H: 507.2091,
found 507.2105.
D. 3-[3-(3-aminopropyloxy])isoxazol-5-ylcarbonylamino]-
2(S)-benzyloxycarbonylaminopropionic acid hydrochloride:
A solution of HC1 in dioxane (4M, 10 mL) was added to
the compound of Example 1004 Part C (0.81 g, 1.60 mmol)
dropwise. The mixture was stirred for 3 h at room
temperature, at which time a white solid had
precipatated out. The product was filtered then washed
with ether and dried overnight under vacuum (0.58 g,
82%): 1H NMR (300 MHz, DMSO)b12.83 (s, 1H) , 8.98 (t,
1H), 7.88(bs, 3H), 7.64 (d, 1H), 7.34 (m, 5H), 6.83 (s,
1H), 5.03 (s, 2H), 4.31 (t, 2H), 4.27 (m, 1H), 3.40 (m,
2H), 2.92 (m, 2H), 2.04 (m, 2H); MS (CI-NH3) m/e 407
(M+H)+; HRMS calc'd for C18H22N407 + H: 407.1567, found
407.1553.
E. 3-[3-[3-(imidazolin-2-yl amino)propyloxy]isoxazol-5- }
yl carbonyl amino]-2-benzyloxycarbonylaminopropionic
acid hydroiodide: The compound of Ex. 1004, Part D
-144-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
(0.42g, 0.95 mmol) and 2-methyimercapto-4,5-
dihydroimidazole hydroiodide(0.35g, 1.42 mmol) were
combined in pyridine (2 mL) and heated to reflux for 4
h. Additional 2-methylmercapto-4,5-dihydroimidazole
hydroiodide(0.35g, 1.42 mmol) was added and the heating
was cdntinued for 2 h. The pyridine was removed in
vacuo and the residue was chromatographed over silica
gel (25.g, 9:3:1:0.6, chloroform, methanol, water,
acetic acid, lower layer). The product fractions were
combined and the solvent removed in vacuo. The residue
was taken up in DMSO (1.5 ml) followed by addition of
methanol (3 mL) then the slow addition of
tetrahydrofuran (40 mL). The mixture was stirred for 30
min then filtered to provide the title compound (0.19g,
34%) as a tan solid: m.p. 201-2 C (dec); 1H NMR 10.14
(bs, 1H), 9.64 (bs, 1H), 8.87 (bt, 1H), 7.34 (m, 5H),
6.78 (d, 1H), 6.75 (s, 1H), 5.00 (s, 2H), 4.22 (t, 2H),
3.85 (q, 1H), 3.56 (s, 4H), 3.55 (m, 1H), 3.23 (m, 3H),
1.92 (m, 2H); MS (esi) rn/e 475 (M+H)*% HRMS calc'd for
C21H26N607 + H: 475.1941, found 475.1942.
Using the above procedures and modifications known
to one skilled in the art of organic synthesis the
following additional examples of Tables 1-5 may be
prepared.
-145-
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
N
a%
co
~' =-'~
fL ~i O~
lf1 eM
O N u u
N O N
0 N N y i 6
Q
m
Q
=Z
O
at x x x x x x x x x x
N
~ a
~+ v
r~
O
i
E
N EI ri ~ M ri ri ri r~ ~ rn rn
qzl
~ =~ ~ r
Ilr -,-r -.4 =H =~
~4 ~4 ~4 ~4 ~4 ~4 ~-l ~, ~4 ~4
~ ~ ~
' o 0 0~ o 0 0 0 0 0~ o ~ ~ 0 0 0 0 0 0 0 0 0 0
W ~ 0
w
'rl ~ -~ z3 10+ ~ z3 - ~~ ~ . ~ ~ =H z3 ~ 'O =.~
~ ~ J. ~ .~ co .~ ~ ~ cru ~ cEid
f~-1 >7 S~-1 ~, ~ >, f ~ -1 ~, S ~ -1 ~1 S ~ >, SM-1 >r-I I f ~ d ~, S ~ d . ~
~t 5~ ~ 1 ~7
y~ y y ~ y y~ y~ y y~ y y
~ N d) N ~ N d N ~ N ~ N N N U) N d N N N
y , 4.) y y 4.) y y y y y
W I Zi I =-1 N ('1 ~ ln l0 L~ CD Ol '-I
~4~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
Ln
~
eM
=
~
~+ ~Jt ~Jt yt =14 =$4 ~i
N a a N ~ =
N
6 f: W U N v v N N N 'y'
cli
0 0 0 0 0 0 O O O O 0 0 O
U U tn am mx txn u] vx~ U Ux Ux U Ux Ux Ux
x x x x x x x x x x x x x x x
M f1 M m m m (n n'1 P7 M f'M M M M P7
tj 'Lj TG R~ ~ ~S Tfy ~ 'LS R7 R7 Z~ ~S ~C
=ri =ri =ri =.i vi =ri =r~ =ri =ri =rl ='4 =rl =ri ='4 ='i
6 E E E E E E E E E E E E E
.,i .'~ .'~ -'~ -'~ -'~ =~ ~~ ~ -~ -~ =~ =~ =ri =,i
Sa Sa $4 $4 ia Sa ~ Sa ?a ~+ Sa ~a S~ Sa Sa
a ~a a a ~ a a a a a u ~
O o 0 0 o O o 0 0 0 o O o 0 0 0 o O o 0 0 o O o O o o O O o
s4 ~
s~ s~ 1 14 ~ 0 s~ 0 t~ 0 q s~ 0
~ =~ ~ =a z1 =~ *C1 =H *c1 =~ R7 =~ zS =~ z'S -H z1 =~ ~ =,~ ~ = ~ .~ = ~ ~ ~
qy ,~ =~ =~
~ tE0 tE6 ~~'. ~ ~~". tEC C ~ cEd cis Cd cEd
$ ~-1 Y~I ~-~ ?i $~-1 >, S~=t >, S~-~ i~-I >, 5~~1 >, 3~~1 yr-i , Nr-I S~-1 >,
}~-t ?i S~1 5, $=1
m y,
J-1 ~ 11 i 11 i S1.1 JJ i J~ i d-1 i 1J }.) .N i y1 ~ 4J i 1J ~ y.l i y.t i
d N d N ~ N 4) N 4) N N N d) N 4) N N N d N 4) N 4) N Q) N N N N N
1) i .L.l y i 1) i 1~ i 1J i 11 ~ 11 ~ L ~ 11 i ,+.) ~ 1J iJ , 11 i JJ ~
= '-I N M cf~ !n l0
r N 01 0 '-1 N m v ln
N N N N N N
14r+
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
~ Ln ~ M
M
E~3 Er3 w ~ ~i o~a ~ a N r' ~
cq cq N ~ o 0 0 0 0 0 0 o
x x ~x U.y~ Ux U U~+ Ux Ux U Ux Ux txn
x x x x x x x x x x x x x x x
~ .=i . i . i .i .i .i .i ~ ri .=t .-i .i .i .i
M M M M M M M M M fn M M M M M
.~.{ .~
~. ~.
.~ .~.{
N N N N N N N N N N N N N
Gi =ri =ri =ri =ri ='i =ri =rl ,ri =,-I ,ri ,rl ,ri =ri
=rl ~=rl r-1 r=i ri r-1 f-1 -i 1-1 -1 ri r-I ~--I ri ri
J1 E, r~1 E+ O O O O O O O O O O 0.8 O O O O O O O O O ~ O O O O
f0 ~[d N F' NNF' N G~' N N~' N~" N .f'. N'6.~' N 0 ~Ny[~ N~a' N~'
>, 7+ ro ~ !0 r=; 8 ~ z -ri ~ ~ ~ *~ ~ ~ ~ !~ ~ ~ M -ri M
1) N ~ J-1 N ~ >t ~ ~ r-i -rS1 >t E --1 >t 5 ~ 5 >t -~ >r =r~1 >r -~ ~t -rrl
~+ =r~i yt =r~1 ~t ~ ~
W 01 O .-i N m ~ 1.t) %O CO 4m O
N N N N M M M m M M M rn m
M c}~
T 0
CA 02221980 1997-11-24
W O 96/37492 PCT/US96/07646
>+ ~l $4 ~ ~o a a a "' I'
$4 ~4 ~, l1] m
v v v
cla N N
N xN x
CV
U U (>
O O 0 0 0 O O O 0 0 0 0 0 0
Cxn Uo cl) CI1 tt1 Ux Ux Ux U Ux Ux C) Ux U.y+
x x x x x x x x x x x x x x x
.-~ =-1 =--~ .-i .-I .-i .-t
M M f~'1 f+1 M M fh f'1 M f~1 f~I (~1 f~1 M ~
.~
~
.r{
~.
.~
i t i i ~ i i i i ~ i
N N N N N N N N N N N N N N
~
V. o 0
r-i
N G.' N G' N N .~'. N .f'. N[S N f.,' N L," N0 N0 NN C,'' N .(". N G.' cEd
-ri *9 -ri fIS -ri !IS -ri -ri ~ -ri ftS -ri (iS -ri ~ -ri (a -ri ts -ri ftf -
r-1 ((f -rl fs -ri fa ri
18 '"~ ~ ri E td ~ ~ ~ ~ ~C ~ . 1 l~6 ~ (5d ~ ~ =ri l~d -~ ~ ~ ~ ~C f~6 ~ ~ !O
(Ed t~i ~
e ;, ~ r-i1 ~ ~, ~ ~, ~~'I ~, ~ ;, E ~, E ~ ~ ~, ~ r~l E E 7, ~ N
+ ~--I N tn eM ln l0 07 01 O -I N m lf)
d+ V~ V~ d~ ~H V~ cp V~ d~ t!1 U) tn tn Lf) ln
49
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
o-
OD
.~
Ln
pq N U CJ rM N f~q
W rn u ~ a
O O O O O 0 0 U.~+ ux mx t~n aj v~j
x x x x x x x x x x x x x x x
o O o o C. o 0 o O 0 0 0 0 0 0
a~ w ~ ~ w v~ ~n a ~n ~n ~ w w d+
~ ~ ~ ~ ~ ~ ~ ~ ~ =~ =~ ~ ~ =~
'C7 'CS ZS *Ci 'CS 2'S 't1 'C 'LS 'CS *C '~ 'CS 'CS +~
S -5
., ~ ,'~ =~ -+ -r+ =,+ 'H
~a $4 H ~4 $4 ~4 ~4
a a a
14 H ~ ~ ~ ~ ~ ~ ~ ~
,~ ~ ~ ~~ ~ ~ ,`~ ~~ .~ ~ ; ~ ~ ~ ~ ~ ~ ~ ` ~ .~ co -i
i~ ~ ~t ~, ~+ ~r $4 ?1 $4 >, H >, >, ~ S4 7, $4
~, 3~ ~, ~+ ~,
ll ~ 1.1 i y) i 1J ~ 1.J ~ 1) i 1J i 1J ~ 1) i 11 i 1) ~ y ~ i
d N N N N N N N w N N N N N ~ N N N N N N N d N N N N N y) N N
4-1 1 1) ~ 1) i 1J ~ 1) ~ 1J y] 1) 1) 1) , 1) . 11 , 1J ~ JJ . }J
tD N n1 sfl in t0 I, c0 m 0 N ln tf) in w l0 tO 1D l0 tG lp %D tO \O (-
CA 02221980 1997-11-24
WO 96137492 PCT/US96/07646
N
r
=
$4 ~4 ~ ~ s~ a a r a r
N N N
cr 3 6 6
N cl
CJFj N N N N p N
~ 0 0 0 0 U O O 0 0 0 u p J. q 0
z 7~ z 2 z x ~ r~=~
x x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
v -0 ~n v~ w r w w
-r1 ri -ri .i 'i ri -rl =ri rl ri ri
27 ~ *~ z3 43 *cf *C~ z3 R~ 'C1
-'~ -'~ -'~ =~+ =~+ =~+ =~+ =~ ri =~ ~
E E E E E E E E E E E
=H -,I -rj -rj =~
~i f=1 ~i ~1 ~i $-I ~i fd $d $1 f-I r r r r
>1 ~t >+ >r ~r f N N N N
a a a a a a a o, a r r r r
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
r N r s4 $4 $i f4 0 14 k $4
~ =~+ =.~ =r~ =~
R3 -4 43 = 4 Rl 1+ ZS =H 'H 23 =rq rq 43 H 10 - t z3 r-i Z3 - i r-4 , i r-I ,
i
RS Si (d .Li m k RS .ti QS ~a Cd .~i RS N N 8
N Ci N L:
f~ rl RS e-1 f0 ri t0 rl Cd e-i Cd -4 Q{ '-i t0 ri f(j -i Cd r-i (o
ri =rf =ri c~ =ri =ri
S i ~ N>, S-4 >, S-i 71 i-i ?r S-l y, $a >, i-i >, S-i ?r S-i >, S i >, ~ E ~
E *tS E ~ E
JJ r 4.) r 4.I r 4J r .L.> r 4J r 1) r Y r 1.1 r 1) r 1) r =r1 f0 -ri RS =rl
(~ =rl fd
Q) N N N d) N () N d) N d) N ~ N Q) N d) N Q) N N N E -1 E+-i
y1 r 1J r y) r 4.Y r 4.) r 1J r y) yJ r y) r 1J r 1.1 r =r-~
>t =ri .>t =ri '.~f -rE{ ,y1
= rl N M eM lf) l0 (M m 0 -4 N M W lt1
t~ r- r r r r r r- r co ao ao ao co m
~ J~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
xM
.=-1
M .~
x
~ x
$4
N U U M N
N N `-' N
8 0 ~ 0 0 0 0 0 0 0 0 0
C) Cx) U U Ux Ux U ux~ txn vx~ vx) ux~ CxI~ U
x x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 0 0 0
.n .r w w n w n ~r ~n n d+ w w w ~
~ ~ ~ ~ ~ ~ ~ ~ ~
N N N N N N N N N N N N CV N N
.r{ .,~ =~-1 ,ri ,ri ,~-I =ri =r{ =r{ .,~ =,~ =ri =rl =ri =ri
O Q O O O O O O O O O O 0 O O O O O O O O O O O O O O O O
Ny Q N NN.~'. N cNt~N N-{..' cN~ N(~' N.~'. cN~f'. N.{ N G." ~Ny
6rO E8 E ~ E ZS E'~ E'~ ~ R7 6~~ ~Ei z3 E
~ 6 r+ E ,-~ @ - E +- -~+ E ~ E -~i E -+ ~ E ~ 6 ~ E ,~ E ,~
=~ >1 =rl ,'~t rl =r1 >t =rI .17 >t -'i .'11 ==i >t ri 'Jt -ri ,1t ri >1 -H >q
-H >1 -='1 >1 rl ,1t
o
~o r co rn o . i N m w in %D r co rn o
co dD co co rn m Om rn Orn rn m Orn Ol Ol .-1
i s2
CA 02221980 1997-11-24
WO 96l37492 PCT/1TS96/07646
= M
ui
n
.,I
a S7~ i~s
~
N W ~ r-I
M eM
N N N N 0 N N N
O ~ O O O O V 4J 0 0 0 U 0
x x x x x x x x x x x x x x x
O O o 0 0 0 0 0 0 0 0 0 0 0 0
~N d~ eM eM m r=1 rl r'1 m M M
' ri r. i
~ 10
.~ ,5 rz E ~
=rl ri ='.{ -~ ,~ ='õ~ ,~õ~
N N N N N N N N
~ ~ ~ ~
O O O O}~ O O O O O O O O O
~ ~ ~ ~ .-~ ' i .-~ ,=-a 21 rOi ~ .Oi z3 ~ ~ =~ ~ rOi ~ rOi '~ ~
N fQti' N 0 N0 ~i. N~.~.,' N00 N N0 N0 0f [EO Sa.~+ fE6 ~ ~~ ~ CE d ~ f0
~ " H ~ c~ td ,-i RS +-a
't7 E~ '~ E *~f E'CS E E~ ~CS E TS ~O E >, ~a >, >, t+ >, ?i $-i >,
=rl r'e C0 -rl t0 =rl tRS =ri tO -rl fI3 =.l cIS 1J i 1) 4.1 ~ 1) ~ 1) ~ 4.~ ~
~.1 ~
=~ ~ -.E=1 ~ E St ~~ ~~ E~t ~~+ ~~r i.~) N 4~.1 N .L N d~l N a.~) N I b ~j N
J.~) N
' ==-I N fn ~ t1) tD Oo O\ 0
-i
0 0 N f~1 eõ t1)
~I .-1 .=i .==I =-i .-1 ==-I ==-~ =-i -i .=i ===1 -1 =-1 -=I
v53
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
~
O
r-i
x!=1 ~
' ~ , 'd R3 93
rn ~ =y.~.~ =f.~j =y.~j ~
u V N (h ~'Ni ~A ac a a W ~N''
N N N N N N cli
O O O O O O O O O O O O
x Ux Ux Ux U uxl vx] Uxl ux~ txn U2 U Ux U Ux
x x x x x x x x x x x x x x x
0 o 0 0 O 0 0 0 o O O 0 0 0 0
m ('~l M fn m M m fh fh Y~1 fn ('7 fr1 m
f1
ri ri ri rl ri ri ri ri rl ri =rl r=1 ei rl rl
~ ~ = 5 s E 'r '~ ~ ~ ~ ~ ~
=H ='i =rl =rl =ri =rl =.i ='i =ri =rl =ri =ri ='1 =ri =ri
s4 ~4 $4 $4 $4 ~4 ~4 ~-f 1.1 s~ ~ w -f
a >1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0~ o 0 0 0 0 0~ o
$4 ~ s~. s4a s4 Wa Wa
~CS ~ 'Z3 =~ ~ -g~ ~ ~ ~ ~ =. i ~ *C3 1 =~ z3 =, ~ 'd =11 1r'S ='1 -~+ z} H
*LS =~
cEd ~ c cEC .~ cEd ~ ~
c6 ~ [d - S ,-i co -+ td ,-t t0 1-i ttf .--i c0 -4 S .-a tti ,r-i Rf -4 rd
,1 R3 ,-t M r-i n3 r-i
S-1 ?+ i-t 71 S4 >i $4 ?1 N>+ S-1 ?1 $-I 71 i=l >+ S4 >r N>1 N>1 $4 >+ $4 ~>+
$4 >+ S4 7r
43 i J3 ~ J..1 ~ J-1 i .L.1 ~ J.) ~ y ~ yJ , yl ~ y ~ y i y i y ~ ,1, y i
N N N N d N ~ N d N N N N N U) N N N N N 0 N d N 4) N Q) N ~ N
JJ I J=1 ~ J-> , -W , 1J , J-1 , 4J , 1J I L , d.) , 41
~ J-) ~ JJ i y ~ y! i
t0 OO O) O =--I N M t() l0 [- cD O) 0 .-I ~==I rl =^i N N N N N N N N N N m
=--I r-I ==i =--I 'i =-i .-1 ==-I ==i =--I =-i =--I '-I ==i =-i
~~~
CA 02221980 1997-11-24
WO 96/37492 PCT/1JS96/07646
y
~ .=I
~
tM `
r=i ri e-1
r=i ri .'s~
=~ ~ ~ ~ ~
a a
~ ~ ~ ~ =n ~ _~ M
LO xw
v v v
M M M N U U
N fV N N 0 N N N 4J
0 0 0 pq 0 0 0 O 0 0 0
2 Z aC ~U ~ 2 Z 2 Z z z
x x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
P7 rn (n m m M m fh fn m M m m M (n
C'a ia" G' G' C.~"
q:l 10 q~ 10
E -~ ,~ ,~ ,~
.,{ .~.{ -~ =~õ~ ,~
S-1 $4 $4 S4 ~i
>1 >t ~ ~t N N N N N N N N N N
~ ~ i i
O O O O O O O O O O 0 C G; t"., 0
t~ G s G t~ G =I =.i -~, =,~ -,~ - ~ = ~ õ~ ,,~ ,~
*t1 =.~ t =.~ 43 ~ ~ ~
>+ Fcc >+ E ~ E ?i E ~ E O O O O O O O O O O O O O O O O O O O O
~ fC .tr" CO f{3 Sr' R$ N 'L." N F." N~.' N N~ N .r.~ N(,~"' N(i N N
qf ri -i fd e-i fRS ri fo e-I fd =ri =.1 m =rl =ri ct~ -ri =ri c~ =ri =r~l ct~
=~-i =ri
$=~ 7, $~=~ ?1 f=+ S-i >, S=4 >, z.7 E 4 E *CS E 4 E'L1 E 9 E z3 E ~ S E
.7-1 i 4.) ~ y) ~ 1] i 4J I -ri (d =ri (~j -ri f~ =rI f~f ='{ ftS =~=i f6 ri
ld 'i !lS =rl IlS =ri ftS
N N d N O N d N N N E-4 6-1 E.-i Er-{ Er- &+-+ E.-i E-4 E-i E.-{
4j , 4.) aJ 41 .u =a >, =ri >, =H >, =-1 >, -r+ >, ~ >, =~ ?, ~ >, .~ >, =~ >,
~ =-i N r'1 cf' U) \O [- N Oi O .-i N M ~M lf1
f~1 f'1 fn fn m rn (*1 V
=-i ~-1 .-1 .-i .=1 '-I ===I .-I ===I =-i ==i .-i .=i .-i ~i
1~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
. rt
~
M R3 Z~ R~
~ 'CS ~ 'CS ~+ =~+ $4 a
cr) =,, rj a a ~t~t~t
M N 44 W W N - - N
O O 0 0 0 0 0 0 O O O O O O O
U U Cp Cll U~ tl~ Cp Cll U U U U U U U
O O O O O O O O O O O O O 0 0
fn m m m M m fn M fn m f1 m m m m
= ~ ~ ~ i ~ i ~ ~ ~ ~ t ~ i
N N N N N N N N N fV N N N N N
.r{ =~.{ =r{ =~ ..~ =r{ =ri =rl =r{ =.i =rl =rl =ri =rl =ri
ri e-i ri r-i r-1 e-i r-1 -1 ~--I r-i r-1 ri r--I r-I -I
O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O
N~y F' Nct} .r'.~ N N N N N'{..~' Nct~ 6," Nc~ N~..' N G."
E z3 8 *O S ~O E ~ E 9 E 6 6 8 15 E 21 E1
& `z3 E E
~~y=~ ~~ E~- ~ i E~-~ i E ~ =E~ E~ E~ 6~ E~ E'-~ + E~ E~ E~- mi ~ ~
=1'7 ~, =="~ ~+ ri ~, P'I = r'~ y, =="~ ,, =/"1 >, r'1 ?4 =~ >, =r'1 ,, =r'1
r71 =~ ?1 -''1 >, =-I ', >,
D l~ OO 01 0 ---I N m V) l0 CA Ol O
101 lf) Lf) tA U) Ln t!) U) U) Ln Ln l0
=-i -1 =-i =-I -4 -4 ri 1-1 =--! -4 ri -4 I1 -4 .i
`~
CA 02221980 1997-11-24
WO 96/37492 PCT/[IS96107646
~
N = ~
OD OD
v
f=i r=~
N
0 ~ N O O O N O O O .[~
x x x x x x x x x x x x x x x
0 0 ~ -i .1 ~ o 0 0 0 0 0 0 0 .a
m M M M M M ~ ~ sr sr M M M M M
O O O O O O O O O O 0 E ~"i ~'i ~'i ~'i ~'i ~'i ~'i ~'i Li ~"i Ci G~" =ri
E E 8 E E (s
~-coi ~-~-1 'r-ij~
N
='i N N N N N N N N N N N
e-1 r-I
O O O O O
N C; N(~. ~-I ri rt .=i rl ri ri -ri rl -.-1 ri ri
'L~ z1 N
'L7 `~ 'C3 ZS 'CS 'C1 Z~ 10
L
=,i3
ri ~ ~ Sa $a S4 Sa Sa a N fa y 4 S4 ~ 'f0
Ei
N M V~ U) l0 C~ OD O) O .--1 N M
r r r r r r r r m co 0o ao ao
=-+ =1 .-i ~ -i -i .1 .1 .1 .-i .1 .1 .-i .i .i
15r4
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
QN N N NQ N N N N N N fV N N N ~N
U U2 U) C) C) Ul t!) U 0U VI CA U 0U Ul U]
1-I ~i 14 O 0 O 0 O O O O ~-i =-I rf .i
m ('7 m rn fn m fn fn m m fn
r-I ~-i rl
Q 4 O O O O O O O O O .'~t .'~7 .~t
~ ~
~ =4 =~ =~ =~ =~ H =~ =~ =E =~ ~ t0 l0 1O
'~-I '-1 r-1 r-I r=I '-I rl e--I rl r-I
>1 >1 .yt >f >f .79 >t >1 >t >t
i i ~
N N N N N N N N N N N i S-1 f-1 id
~ i N
ri r=I
N N N N N N N N N N N 0 .~i 8 0
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
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CA 02221980 1997-11-24
WO 96137492 PCT/US96107646
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
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WO 96/37492 PCT/US96/07646
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WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96137492 PCT/US96107646
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WO 96/37492 PCTIUS96/07646
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CA 02221980 1997-11-24
WO 96137492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
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ri f - 1 cC -=-I cz =H cn =r1 0 =.i c0 -rl t0 =H Rf -li M -+ Rf =ri cd =ri t6
1J
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i ~t
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-0 '0 Lf) lf1 lt) U) lf) N
Ln Ln Ln N LO In N Ln LO LO Ln Lr) U) N LO
eq.4
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
.~ .~
Ln v 0
ri N pq
N U U U ` V
N Li W
N N N 6 6 .T. V
m o 0 0 0 0 0 0 (o' o 0 0 0
4-1 Ux Ux Ux Ux Uy+ Ux Uy+ Uy+ Ux v~ u) v1 vJ
x x x x x x x x x x x x x x x
O o 0 0 0 0 0 0 0 0 0 0 0 0 0
=y~ ~ V~ ~ etl V~ ~ ~ d~ V~ ~ ~ cM d~
e=i rl 'i .i rl r1 ri rl ri .{ -ri 'i .i rl rl
T$
=-+ .~ =~ =~ ..=~ =a .~ ..~ .., -.~ ..~ ,.~ ..~ -~ ,i
E E E; E E E E E; E E E E E E E
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>1 >1 >1 >1 >1 >1 >1
a a a a a a a a a a a a
0 0 0 0 0 0 0s~ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 a0 0 0 0
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.i ~ -~i
.~~ ~ .~~ ~ ~~ .~~ ~ ro ~ E .~c6d m rEd lc=-o
cd r-i M r-I c0 r-i 0 -= 1 Rf +--i nS 1-1 a{ .-i m ,--i Cd -4 f ,-i M 'A cts
r-I co r-I Cd r-4 RS ~-q
S-l >1 k ?i $t >+ S4 >1 k >+ 74 >+ $a >1 $4 >1 $-r >+ $4 yv W ?i N>r Sa ?1 k
>i S I >i
1J ~ J.l ~ 1> ~ 1) ~ 1) ~ 1) ~ J~ ~ J~ i y i 1.1 ~ y i 1J ~ y ~ y t ,y.1 i
N N N N d N N N N N w N 0 N d N N N w N d N d N N N N N d N
1) ~ 1J i JJ i J~ ~ y ~ y1 ~ 1! ~ JJ i y ~ J.J , 1J i y 4J
~ l0 [~ OD m O .--1 N !1 V~ t!) lD l~ OD Oi C.
t!) tn tf) ln l0 w kO kD l0 w tD tD l0 %D [~
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1qS
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
{
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u~
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ux] a] Ux Ux Ux Ux Ux Ux Ux Ux Ux Ux .u Ux
Qi Qi 2 '7r '~r z 2 ^lr 'Tr x z 2 2
x x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
w ~r ~n ~r ~r w w ~n w ~r ~ ~r ~ w ~r
~ 'CS 23 R~ 'LS ~ R7 *~ R7 R~ ZS
õq
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=rl =rl =.i =rl =ri =~-1 -ri =ri rl
S4 $4 1-i ~1 N S4 S4 $1 $1
O 0 O O OO O O O O O O O OO O O O 0 O O O q ~ O q
9 y~ O ~t r $a q Sa G y~ q y~ O }.~ G G ~ ~ ~
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=~ ~ ." Rf ~ f~ ,L'. t1S .Gi .tr' cC .L~i RS .fy' (6 .Li t0 fiS .4 Cd N
co r-I nS ,-i td ~ cti r=i tC .-i co ~-l c0 11 rt -i co -4 cd =rt -~ S =rq
({} ~
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o ~ $ ~
1~ ~ 1) 11 1) ~ 1) 1.1 ~ JJ y) i
d N N N N N d N N N d N N N ~ N d N N N N N =.~., rl Ei e-i -ri E ~
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~ =ri =.i ~ ~ >1
N f`') d~ Ln lO r- W 01 O 1-1 N m sT Ln
r r r- r r r r r r w w w co 0o co
ln lf) ll) N U1 U) U1 Lt) tn tf) tn ln tfl ln Ln
=
tq~
CA 02221980 1997-11-24
WO 96/37492 PCT'/i7S96/07646
co
xM
(,) rli
w r' a u w~ v
0 0 0 0 0 0 o 0 0 0 0 0 0 0
t~ u c~ u ~ u u c~ cn u~ uz cn ~n u~ u
x x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
r ~ ~ ~ r ' a w ~ ~ w ~ w =a~ ~r
N N N N N N N N N N N N N N N
.'{ .ri ,ri ,~.{ ,~ .~{ =~õ~ -~ -~ =~ ~ =~ ~ =~ =~
e-i r-I rl r-I rl r-I r=i ~--i r-I rl ~--I e-i ~--1 ~--I e-i
O O O O O O 0 O O O O O O O O O O O O O O O O 0 O O O O O O
C." N~..=' N
~-i -r=I =rl (a r=I -ri =ri - =ri =ei ='i -ri -ri =.I
s s 5 '0 e =~ri -riE ~e s.i 5 E E 8
-=~ nS =H cIi -rl crS -i cC -rq cd -ri co -.I co -,I Rf -rt fO =H co =,i td
=ri to -.j td -.1 tTS -i RS
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co ao co co rn m rn m m m arn o, rn am o
LO Ln Ln LO Ln Ln Ln Ln Ln Ln U) LO U) Ln
~
1~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
t0>1 'd s-l ~, $4 .,~ , ~
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N N N
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x x x x x x x x x x x x x x x
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sM d~ V~ d~ V~ ~ W M f"1 fn m f'7 m
f'1
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N N N N N N N N N a a a W a
O O O O O O O O 'J O O O O O
=,j =,j =H =,i =H S-I G~i 14 C' }.~ L Sa~~ La
`CS =.1 *LS -~i 'O -~ 1CS -.~ 'LS =~i -~i ~ -a
O O O O O O O O O O O O O O O O ~ E >+ E ?, r >, E ~.~F. ~ ~~
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ri =ri .rl =ri -ri =ri (d =ri r-1 ri i~ r-1 ld ~-i ~-i ta i=-I ri
=~.. ~ =ri~ ~ ~.. ~i .17 f{f=I .~t ~1 .17 f-I ~ ~-Ifd.1t ~d .1t .7t
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=-i N (=) ~ l() \p r m p% O .-I N M t!)
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%D l0 tp lp %D %O %D l0 t0 %O %O %O %D l0 kp
A9$
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
M
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M
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u V ~ M !Mr! N
Li CJ ~J N ly n7 N N N ~ ~ -
~ O O Qv CU7 ~
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x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
M m PM m M M m M m M M M M M M
ri ri ~i =ri ri ri ri ei ri =ri =rl -rl ri =rl ri
w~i -e-I -ri =rl =rl -rl -ri =ri =ri ==-1 -ri ='1 =ri =~i ='i
F. E E E. E F. F. E Ei P. Ei F.. F. F.
"~ 'rl =~=1 -rl =ri =ri -='i =ri =ri =ri =ri =rl =ri =.i =r1
1-1 $4 ~I $4 f-I ~4 $1 f-7 $4
>7 >t .~t ~7 ~t .~i .~7 r~7 >7 .1t
a a a a a a a a a a a a
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
~+ ~+ q y.~ ~ sj r~ sj s4 q $4 q G ~ ~ w ~ w r.
'C3 *~ '0 r, 'L7 ri 'LS =-1 75 =14 'L3 ='-I 'tj _4 10 =~ 'Cf =.-i 10 gi 'CS -
t 'LS 1 ~ =.-i `CS -~ ~LS =~
c~d ~ .~ ~ ~ ~ ,~ ~ ~ ~ .~ c~d ~ t~6 .~ tE 6 .~ cd 4 ~ rEC ~ c~d ~ ~ .~ ct
c0 -q S .-i co r-t cu r-t m ~H nS r-+ ca r-I c0 ,--i id r-i td '-t (6 r-t RS
,--I RS .-i
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1) ~ 1J 1) ~ 1J 3-1 ~ 1) ~ JJ 1J 1J i JJ i 1J 1) ~ yJ ~ 11 i 1J i
N N N N ~ N d N 4) N Q) N 4) N Q) N N N Q) N ~ N d N d N N N N N
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\O l0 t0 l0 t0 %O l0 l0 t0 lG w %O t0 LD l0
. ~~~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
= '^ -
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cq M N U U
N N N O N N N ~ 4J
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x x x x x x x x x x x x x x x
0 0 0 0 0 0 0 0 0 0 0 o O o 0
P~f M1 f~1 l~'1 M f'~1 M f7 P7 f~'1 fh f~1 f') P1 N1
~ =~ ~ ~ =~
N N N N N N N N N N
O O O p O O O O 0 O $4 O ~.{ '~S =rl 'CS =.~ ~ rl '(} =rl ~} =.~ e-i ri ,--~
ri r-I -1 ,-~ r-1 ,-=1 r-1
>+ E >, E >1 E ?, E >, E O O O O O O O O O O O O O O O O O O O O
" fd .t=~'' (C ,4 i f0 4 fli .~S (0 N 0 N G"., N~.' N 0 N(.,=' N 0 N .{~., N
={-i' N(.i N[~'
[o r=i ({j r-I (a r-I f0 e-i -ri ~y -ri ~} =ri ~y ri r~ ='i =rl c~ =ri c~y =rl
QS =rl ccy =.i
$+ $4 ? i4 E ~3 E'O E'D E'c3 E z3 EE 10 E t3
~ u y u ~ =~ ct =H cd -~ (0 =.i cC =~ ~ =.i C6 =H c0 =~ Rt -~ .~
N
N NN aD N a~N a) N ~ E-i E-i E-+ E'-+ Er-+ Er-+ Er+ E'-+
4.) I 1) i 1.1 , .I.l i JJ ~ =r=I ,1t =r~ .'~ =ri .~+ =rl .7t -.i >, ri >t -rl
=rl ,1t
.-I N m er tn %D I~ OD O1 O rl N r'1 ln
fh f1 m m th m ('1 m m -W v v
\O t0 l0 t0 tD ~O \O l0 lD t0 %O l0
ao~ ~
CA 02221980 1997-11-24
W O 96/37492 PCT/US96/07646
:
~
M .-. . .
M~=. *~ ~ ~ q5
V .~t .1t .7t S-I $4 =$4 $4
xM
al x a a cq = -0:
M clq a
N (n
6 6 W U m - v ~ N N N
O O 0 0 0 0 0 0 0 0 O O O O O
U U C!~ U) p1 u~ CA U] U t) U U U C) C1
0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
M M M M M M M M M m m M M M M
~ i ~ i ~ ~ i ~ i i
N N N N N N N N N N N N N N N
=,i =ri .~..~ ='{ ,r{ -,~ ,r1 ,rl ,.i ,.1 ,ri =ri =.1 -rl ='i
r-I ~==I r-i e-i r-I r-1 e--I '-i r-i ~--I ~-i r-=I ~=-i e-i r-I
O O O O O O O O O O O O O O O O O O O O O O O O O O O O O O
N .t'. N{..+" N~." N(~' N 0 N{." N .{". N .(". N G'. N i Ni N .t'.i N 0
N'~..~" N
ri =rl =,i
RS .-1 ~ =ri fd =.I fIS =ri Tf~3 -ri fd =ri RS =ri -ri RS =rl Z4 =rl =rl (~ -
rl
~ ~ (~0 .q (~6 ,~ f~d ,~ ~ =rl cEd ,~ ~ ,~ ~ ~ tC ~ t~0 ,~ t~C ~ f~0 =ri ~ ~ ~
ri tEid
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~ 10 -0 c Ln Ln Ln Lr; Ln us U) Ln Ln in %D
to w w w %o %o %D tD
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
co cr%
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14
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a
fV N N N N N N N N N N N N N N
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x x x x x x x x x x x x x x x
M m m m m m ~ ~ m m m m m
CO=' ~O=' GO'~ tO.' LO". L~'~ ~O,' G~"~ G G~'~ 4~~i A -r~"i
=~ ~~ ='~ ='~ -'~ "~ "~ "'~ '~ '~ -~ -~ E
rd E E E E E E E E E E E ~S
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-rq
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!nr,,
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
a a
N N N N N N N N N N N N N
CQxJ O O .q O O O [t O O O O O O
cA UxI U Ux L/~ tx/] Ux Ux tx!~ mx U Ux tx1~ uxl
x x x x x x x x x x x x x x x
-=t =-+ ~I o 0
P~1 M Cn ef~ V~ K1 f'~1 fn M m M m m
O O O O p O O O O O O >r-i , ?,
'E =~ ''i "i -~ =~ =.i -~ =~ =~ tc ~o ~n
c0 c0 E E E E E E E g E =,I
.~ '- ~ i ,~ 1-1 ~-m i cE0
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I I N N N N N N N N N N N =~ =~ =~
$4 N ?-I
t
r-i 1-1 e-1 ~--1 i r-I -I r--1 r-1 N
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E E E E E E E E E E .p 1
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i l0 co O~ O =-i N m U1 1D CO Q1 O
r r r ao m co m co w co co Oo w rn
%o %O ko l0 %D t0 tD lp tp ~o to t0 t0 lo t0
ao3
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
o ~o r
r n
.r
P~G ~ P~0 ~ P~9 P~0
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t0 ~O l0 ~O ~O l0 ~O ~D f~1 n1 [=1 t'1 'W V~
rl 'i ri r-1 'i rl r1 .i ei rl r1 .i 0 0 0
't3 'C~ TCS R3 R~ 'CS ~ '~ '~ ZS Z3 'd N N N
=rl =rl =r{ 'r{ '~.{ 'r~ "r.{ '...~ 'H =,1 =ri '.i RS fz fIf
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E E E E E E E E E E E E E E
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N N N N (N N N N N N N N N N N
N K1 lf) tO L- CO 4m O .-I N M v tn
Q) Q1 O% ON ON m m m 0 O 0 0 0 O
lo %o k0 to %o %o ~O lo \o t~ t~ t~ r r t~
c>,Y1
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
pq
~ .ii M N M N-1 M
1 I a ~ ~
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04
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e-i -1 H
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[o S RS cd =ri t0 co RS Rf (0 co rq .i $a .1 .i ~ 11 -1 ~ O O O O
.[ .O 4 .O >1 4 .O .O .O O ~ =~ -~ =.i =.t
,J 4J a., ji a 4J 4J y L., ,J J-) -1 ,-~ -1 r,
0 0 0 O O O O O 0 0 0 o O O O O O O O
0 9 0 f-~" Gi G G" Ci [." L: N L; N 0 N G' N
-ri -rl -rl "i 'i 'ri ri =ri -ri co -ri
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ctf td RS RS ctS t~ ttS rG S cd S -.i ttS -.i af =~ t0 ~ ca N N N N N N N N
N N N r~ ~r-i i ~ .~i >1 ~~ ~~
l0 [- co m O -1 N m eM tt1 %O l71 tD ~ m
0 0 C. 0 .-~ -=1 1-1 .-+ -+ r r r r
r r n r r r r ~ r r r r r ~ r
~~~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
N2 N N N ,QN N ,SN~ N N N N N N N N
V Q U] p~ U U U U L!2 U t/~ U U] U tJ1
x x x x x x x x x x x x x x x
N N N
N N N N sM V' m m m f+1 N N fh M
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a) ~
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r+ fa ~+ a a r~ a a a a a
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.0 m .~ as .~ ca N r~ q G q 0 q 9:
fif rl m ri co r=I fa rl =ri -ri 4J =rl c~ =rl -rl =ri =ri =ri ri -ri =ri
~>, f i>, ?~ 1 r >, N E N E ~C7 E- E*=i E.-i E+ E-+ E-i
N N a) N N N ~ N Li ~ ~"i ~ =e-i =;i ~, =rl >, ='i >, =ii ,1, =ri .1, =ri E E
E ~ =~ =~ r.{
~' ~ ~' ~ ~~t .L2 ~t =~ N t!1 N Lf1 N tn N Lf1 N Ln N ln N r-{ ~ N~
O~ O .-i N m ~ [, sT lf) l0 [- OC Ql O
t- co co co W W co Q) 01 O) cn m m O O =
r r r r r r r r r r r r r co 0o
CA 02221980 1997-11-24
WO 96137492 PCT/IIS96/07646
= d,
w a a w w
N N ,Nq N N N Nq fV N N N N
0x CxJ t0 Ux C0 t2 Ux c 1~ Ox Ux U Oi Ux C0
CJ2 !) xn
x x x x x x x x x x ~, W~ OQ x x
o O -i -i =-4 -~ o O r+ --+ O 0 O O O
m f'n N CV m m M m N fV d~ W cM CV N
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N
N ~ ~
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P~ f~ G~ LA a) N d) a1 N a) 0 0 =~-i ==-1 ==i -~i N N N N N N N ~ =ri =rl
ft$ (d (t ftS R$ (ts ~ td ~ r=i r-I
O O O O 0 0 O 0 O 0 =rj O -4 O O O O O O
Gi t' S L F G Cti G L: E~ .O t. N N Ci N Li
.~.{ .~ =.={ .~ .~..~ .'.{ .r~ =r{ .~ =r~ =rl =r1 1.) =ri ri =ri
E E E E E E E E E E N E N E ~ E E
-4G co -H ~ cC
=~ ~ ~ ~ i ~ ~ ~ ~ . ~ A(~) co
i e-~ ~ r-1 i ~===I ~ ~=-~ ~ ~ , ' ' .~~1 E eG-~ =rF~ >1
N~ N~ N>t N>t N N N N N N
N n1 eN tfl %O [- c0 O) O .==i N r1 VO r=1 -IV
O O O O O O O 0 =-i 1-I =-i -4 1 N N
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
r
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CA 02221980 1997-11-24
WO 96137492 PCT/US96/07646
1 =
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
. s r
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WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCTlUS96/07646
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WO 96137492 PC7'lUS96/07646
=
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96l07646
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CA 02221980 1997-11-24
WO 96/37492 PCT/iTS96107646
~
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
>1
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CA 02221980 1997-11-24
W4) 96137492 PCT/RTS96(07646
r-4
77
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
.~ ^
tti ~i .y1 .=~ W t0 t0 .C~ N r-~I ~
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44 44 4-4
w 44 44 44 44 w w w 44 44 w 44 m rn m rn m m a, am rn rn o, rn rn rn m
m rn arn m m rn m rn m m rn m m (7) rn '
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CA 02221980 1997-11-24
WO 96137492 PCT/L7S96/07646
.~
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WO 96/37492 PCTIUS96/07646
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WO 96137492 PCTJUS96107646
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WO 96/37492 PCT/US96/07646
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WO 96/37492 PCT/US96107646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96107646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
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a45
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96137492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCTl1JS96/07646
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
x x x x ~ ~ a) CD x x ~ ~ x x x x x x
w a~a o~v o~a ~
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CA 02221980 1997-11-24
WO 96/37492 PCTIUS96/07646
1 -
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t
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
= ln M M N
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i --1 ~-i =--I
CA 02221980 1997-11-24
WO 96l37492 PCT/US96/07646
~
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aS3
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
x x x x x x xCD x ~ x x x x
0
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4kJ 4J~~~
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as~
CA 02221980 1997-11-24
WO 96137492 PCTlUS96/07646
~ pNq pNq N N N ,qN N N N N N N
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
aoav aooa
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CA 02221980 1997-11-24
WO 96/37492 PCT/LIS96/07646
N N
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CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
= c~xxx x x x x x xxx x xxx x ~~ ~
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f~ = =E N N ~F,= =~~ `~U! =E N N =L'i E 0 0 >1 -4
N N L~i
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(1~ =.i ='i 4.) CV J-1 N=rl 1.1 N=rl 4J N=ri =rl L7 N N=ri ri 1) N 4iNri ri
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z =--I ~--I ~--I =--I =--I 'i -4 -1 -4 -i .-i .i -1 i--I .-i .=1 '-1 -I -I
a~ ~
CA 02221980 1997-11-24
WO 96137492 PCT/U896/07646
w
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=~ ~ =a .i Z, .=~
Q5~
CA 02221980 1997-11-24
WO 96/37492 PCT/US96/07646
Utility
The compounds of Formula I of the present invention
possess activity as antagonists of integrins such as,
for example, the aV03 or vitronectin receptor, avP5 or
aso1, and as such have utility in the treatment and
diagnosis of cell adhesion,`angiogenic disorders,
inflammation, bone degradation, cancer metastases,
diabetic retinopathy, thrombosis, restenosis, macular
degeneration, and other conditions mediated by cell
adhesion and/or cell migration and/or angiogenesis. The
integrin antagonist activity of the compounds of the
present invention is demonstrated using assays which
measure the binding of a specific integrin to a native
ligand, for example, using the ELISA assay described
below for the binding of vitronectin to the av03
receptor.
The compounds of the present invention possess
selectivity for the av03 receptor relative to the
GPIIb/IIIa receptor as demonstrated by their lack of
activity in standard assays of platelet aggregation,
such as the platelet aggregation assay described below.
One of the major roles of integrins in vivo is to
mediate cellular interactions with adjacent cells. Cell
based adhesion assays can be used to mimic these
interactions in vitro. A cell based assay is more
representative of the in vivo situation than an ELISA
since the receptor is maintained in membranes in the
native state. The compounds of the present invention
have activity in cell-based assays of adhesion, for
example as demonstrated in using the cell adhesion
assays described below.
The compounds of Formula I of the present invention
may be useful for the treatment or prevention of other
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diseases which involve cell adhesion processes,
including, but not limited to, osteoporosis, rheumatoid
arthritis, autoimmune disorders, bone degradation,
rheumatoid arthritis, asthma, allergies, adult
respiratory distress syndrome, graft versus host
disease, organ transplantation, septic shock, psoriasis,
eczema, contact dermatitis, osteoarthritis,
atherosclerosis, metastasis, wound healing, inflammatory
bowel disease and other angiogenic disorders.
The compounds of Formula I have the ability to
suppress/inhibit angiogenesis in vivo, for example, as
demonstrated using animal models of ocular
neovascularization.
The compounds provided by this invention are also
useful as standards and reagents in determining the
ability of a potential pharmaceutical to inhibit
integrin-ligand binding. These may be provided in a
commercial kit comprising a compound of this invention.
As used herein " g" denotes microgram, "mg" denotes
milligram, "g" denotes gram, " L denotes microliter,
"mL" denotes milliliter, "L" denotes liter, "nM" denotes
nanomolar, M" denotes micromolar, "mM" denotes
millimolar, "M" denotes molar and "nm'1 denotes
nanometer. "Sigma" stands for the Sigma-Aldrich Corp.
of St. Louis, MO.
The utility of the compounds of the present
invention may be assessed by testing in one or more of
the following assays as described in detail below:
Purified avP3 (human placenta) - Vitronectin ELISA,
a.03-Vitronectin Binding Assay, Human Aortic Smooth
Muscle Cell Migration Assay, In Vivo Angiogenesis Model,
Pig Restenosis Model, Mouse Retinopathy Model. A
compound of the present invention is considered to be
active if it has an IC50 or Ki value of less than about
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ltM for the inhibition of avP3-Vitronectin Binding
Assay, with compounds preferably having Ki values of
less than about 0.1 pM. Tested compounds of the present
invention are active in the aVP3-Vitronectin Binding
5 Assay.
Purified av.0 (human placenta) - Vitronectin ELISA
The aVP3 receptor was isolated from human placental
extracts prepared using octylglucoside. The extracts
10 were passed over an affinity column composed of anti-avP3
monoclonal antibody (LM609) to Affigel. The column was
subsequently washed extensively at pH 7 and pH 4.5
followed by elution at pH 3. The resulting sample was
concentrated by wheat germ agglutinin chromatography to
provide gave two bands on SDS gel which were confirmed
as av03 by western blotting.
. Affinity purified protein was diluted at different
levels and plated to 96 well plates. ELISA was performed
using fixed concentration of biotinylated vitronectin
(approximately 80 nM/well). This receptor preparation
contains the avP3 with no detectable levels of avos
according to the gel (aVP3) and according to effects of
blocking antibodies for the av03 or aVp5 in the ELISA.
A submaximal concentration of biotinylated
vitronectin was selected based on conc. response curve
with fixed receptor conc. and variable concentrations of
biotinylated vitronectin.
aVp3-Vitronectin Binding Assay
The purified receptor is diluted with coating
buffer (20 mM Tris HCl, 150 mM NaCl, 2.0 mM CaC12, 1.0
mM MgC12=6H2O, 1.0 mM MnC12=4H2O) and coated (100
L/well) on Costar (3590) high capacity binding plates
overnight at 4 C. The coating solution is discarded and
the plates washed once with blocking/binding buffer (B/B
buffer, 50 mM Tris HC1, 100 mM NaCl, 2.0 mM CaC12,1.0 mM
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MgC12=6H2O,1.0 mM MnC12=4H2O). Receptor is then blocked
(200 L/well) with 3.5% BSA in B/B buffer for 2 hours at
room temperature. After washing once with 1.0% BSA in
B/B buffer, biotinylated vitronectin (100 L) and either
inhibitor (11 L) or B/B buffer w/1.0$ BSA (11 L)is
added'to each well. The plates are incubated 2 hours at
room temperature. The plates are washed twice with B/B
buffer.and incubated 1 hour at room temperature with
anti-biotin alkaline phosphatase (100 L/well) in B/B
buffer containing 1.0% BSA. The plates are washed twice
with B/B buffer and alkaline phosphatase substrate (100
L) is added. Color is developed at room temperature.
Color development is stopped by addition of 2N NaOH (25
L/well) and absorbance is read at 405 nm. The ICso is
the concentration of test substance needed to block 50%
of the vitronectin binding to the receptor.
Inteqrin Cell-Based Adhesion Assays
In the adhesion assays, a 96 well plate was coated
with the ligand (i.e., fibrinogen) and incubated
overnight at 40 C. The following day, the cells were
harvested, washed and loaded with a fluorescent dye.
Compounds and cells were added together and then were
immediately added to the coated plate. After
incubation, loose cells are removed from the plate, and
the plate (with adherent cells) is counted on a
fluorometer. The ability of test compounds to inhibit
cell adhesion by 50% is given by the IC50 value and
represents a measure of potency of inhibition of
integrin mediated binding. Compounds were tested for
their ability to block cell adhesion using assays
specific for avP3, avos and a5p1 integrin interactions.
Platelet Aggregation Assay
Venous blood was obtained from anesthetized mongrel
dogs or from healthy human donors who were drug- and
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aspirin-free for at least two weeks prior to blood
collection. Blood was collected into citrated Vacutainer
tubes. The blood was centrifuged for 15 minutes at 150
x g (850 RPM in a Sorvall RT6000 Tabletop Centrifuge
with H-1000 B rotor) at room temperature, and platelet-
rich-plasma (PRP) was removed. The remaining blood was
centrifuged for 15 minutes at 1500 x g (26,780 RPM) at
room temperature, and platelet-poor plasma (PPP) was
removed. Samples were assayed on a PAP-4 Platelet
Aggregation Profiler, using PPP as the blank (100%
transmittance). 200 L of PRP (5x108 platelets/mL) were
added to each micro test tube, and transmittance was set
to 0%. 20 L of ADP (10 M) was added to each tube, and
the aggregation profiles were plotted (% transmittance
versus time). Test agent (20 pL) was added at different
concentrations prior to the addition of the platelet
agonist. Results are expressed as % inhibition of
agonist-induced platelet aggregation.
Human Aortic Smooth Muscle Cell Migration Assay
A method for assessing av53-mediated smooth muscle cell
migration and agents which inhibit avP3-mediated smooth
muscle cell migration is described in Liaw et al., J.
Clin. Invest. (1995) 95: 713-724).
In Vivo Angiogenesis Model
A quantitative method for assessing angiogenesis and
antiangiogenic agents is described in Passaniti et al.,
Laboratory Investigation (1992) 67: 519-528
Pig Restenosis Model
A method for assessing restenosis and agents which
inhibit restenosis is described in Schwartz et al., J.
Am. College of Cardiology (1992) 19: 267-274. 35
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Mouse Retinopathy Model
A.method for assessing retinopathy and agents which
inhibit retinopathy is described in Smith et al.,
invest. Ophthal. & Visual Science (1994) 35: 101-111.
- Dosage and Formulation
The compounds of this invention can be administered
by any means that produces contact of the active agent
with the agentI s site of action, the avP3 integrin, in
the body of a mammal. They can be administered by any
conventional means available for use in conjunction with
pharmaceuticals, either as individual therapeutic agents
or in a combination of therapeutic agents, such as a
antiplatelet agent such as aspirin, piroxicam, or
ticlopidine which are agonist-specific, or an
anti-coagulant such as warfarin or heparin, or a
thrombin inhibitor such as a boropeptide, hirudin or
argatroban, or a thrombolyti.c agent such as tissue
plasminogen activator, anistreplase, urokinase or
streptokinase, or combinations thereof. The compounds
of the invention, or compounds of the invention in
combination with other therapeutic agents, can be
administered alone, but generally administered with a
pharmaceutical carrier selected on the basis of the
chosen route of administration and standard
pharmaceutical practice.
The dosage of the novel cyclic compounds of this
invention administered will, of course, vary depending
upon known factors, such as the pharmacodynamic
characteristics of the particular agent and its mode and
route of administration; the age, health and weight of
the recipient; the nature and extent of the symptoms;
the kind of concurrent treatment; the frequency of
treatment; and the effect desired. A daily dosage of
~
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active ingredient can be expected to be about 0.001 to
milligrams per kilogram of body weight.
Dosage forms (compositions suitable for
administration) contain from about 0.1 milligram to
5 about 100 milligrams of active ingredient per unit. In
these-pharmaceutical compositions the active ingredient
will ordinarily be present in an amount of about 0.5-95%
by weight based on the total weight of the composition.
The active ingredient can be administered orally in
10 solid dosage forms, such as capsules, tablets, and
powders, or in liquid dosage forms, such as elixirs,
syrups, and suspensions. It can also be administered
parenterally, in sterile liquid dosage forms.
Gelatin capsules contain the active ingredient and
powdered carriers, such as lactose, starch, cellulose
derivatives, magnesium stearate, stearic acid, and the
like. Similar diluents can be used to make compressed
tablets. Both tablets and capsules can be manufactured
as sustained release products to provide for continuous
release of medication over a period of hours. Compressed
tablets can be sugar coated or film coated to mask any
unpleasant taste and protect the tablet from the
atmosphere, or enteric coated for selective
disintegration in the gastrointestinal tract.
Liquid dosage forms for oral administration can
contain coloring and flavoring to increase patient
acceptance.
In general, water, a suitable oil, saline, aqueous
dextrose (glucose), and related sugar solutions and
glycols such as propylene glycol or polyethylene glycols
are suitable carriers for parenteral solutions.
Solutions for parenteral administration preferably
contain a water soluble salt of the active ingredient,
suitable stabilizing agents, and if necessary, buffer
substances. Antioxidizing agents such as sodium
bisulfite, sodium sulfite, or ascorbic acid, either
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alone or combined, are suitable stabilizing agents.
Also used are citric acid and its salts and sodium EDTA.
in addition, parenteral solutions can contain
preservatives, such as benzalkonium chloride, methyl- or
propyl-paraben, and chlorobutanol.
suitable pharmaceutical carriers are described in
Remington's Pharmaceutical Sciences, Mack Publishing
Company, a standard reference text in this field.
Useful pharmaceutical dosage-forms for
administration of the compounds of this invention can be
illustrated as follows:
Capsules
A large number of unit capsules are prepared by
filling standard two-piece hard gelatin capsules each
with 10 milligrams of powdered active ingredient, 150
milligrams of lactose, 50 milligrams of cellulose, and 6
milligrams magnesium stearate.
Soft Gelatin Capsules
A mixture of active ingredient in a digestable oil
such as soybean oil, cottonseed oil or olive oil is
prepared and injected by means of a positive
displacement pump into gelatin to form soft gelatin
capsules containing 10 milligrams of the active
ingredient. The capsules are washed and dried.
Tablets
A large number of tablets are prepared by
conventional procedures so that the dosage unit was 10
milligrams of active ingredient, 0.2 milligrams of
colloidal silicon dioxide, 5 milligrams of magnesium
stearate, 275 milligrams of microcrystalline cellulose,
11 milligrams of starch and 98.8 milligrams of lactose.
Appropriate coatings may be applied to increase
palatability or delay absorption.
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The compounds for the present invention can be
administered in intranasal form via topical use of
suitable intranasal vehicles, or via transdermal routes,
using those forms of transdermal skin patches wall known
to those of ordinary skill in that art. To be
administered in the form of a transdermal delivery
system, the dosage administration will, of course, be
continuous rather than intermittant throughout the
dosage regimen.
The active ingredient can be administered
intranasally to a mammal at a dosage range of about 0.01
to 0.5 mg/kg while the preferred dosage range is about
0.01-0.1 mg/kg.
Compositions of the active ingredients can be
administered intranasally by preparing a suitable
formulation of the active ingredient by procedures well
known to those skilled in the art. Preferably the
formulations are prepared with suitable nontoxic
pharmaceutically acceptable ingredients. These
ingredients are known to those skilled in the
preparation of nasal dosage forms and some of these can
be found in REMINGTON'S PHARMACEUTICAL SCIENCES. 17th
edition, 1985 a standard reference in the field. The
choice of suitable carriers is highly dependent upon the
exact nature of the nasal dosage form desired, e.g.,
solutions, suspensions, ointments, or gels. Nasal
dosage forms generally contain large amounts of water in
addition to the active ingredient. Minor amounts of
other ingredients such as pH adjusters, emulsifiers or
dispersing agents, preservatives, surfactants, jelling
agents, or buffering and other stabilizing and
solubilizing agents may also be present. Preferably,
the nasal dosage form should be isotonic with nasal 35 secretions.
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An example of a nasal solution composition of this
invention includes:
Active Drug 0.2-2
Sorbitol 0.6 g
Benzalkonium chloride 0.002
H drochloric acid to adjust pH
Sodium hydroxide to adjust pH
Purif ied water to 10 mL
In this example the active drug can be in one vial
and the rest of the formulation can be in another vial.
The drug can be reconstituted when needed.
The formulation of this invention may be varied to
include: (1) other acids and bases to adjust the PH; (2)
other tonicity imparting agents such as glycerin and
dextrose; (3) other antimicrobial preservatives such as
other parahydroxy benzoic acid esters, sorbate,
benzoate, propionate, chlorbutanol, phenylethyl alcohol,
and mercurials; (4) other viscosity imparting agents
such as sodium carboxy-methylcellulose microcrystalline
cellulose, polyvinyl-pyrrolidone, polyvinyl alcohol and
other gums; (5) suitable absorption enhancers; (6)
stabilizing agents such as antioxidants, like bisulfite
and ascorbate, metal chelating agents such as sodium
edetate and drug solubility enhancers such as
polyethylene glycols.
The above formulation can be administered as drops,
sprays, aerosols or by any other intranasal dosage form.
Optionally, the delivery system can be a unit dose
delivery system. The volume of solution or suspension
delivered per dose can be anywhere from 5 to 400 L, and
preferably between 50 and 150 L. Delivery systems for
these various dosage forms can be dropper bottles,
plastic squeeze units, atomizers, nebulizers or
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pharmaceutical aerosols in either unit dose or multiple
dose packages.
The combination products of this invention, such as
the novel aVP3 antagonist compounds of this invention in
combination with an anti-coagulant agent such as
warfarin or heparin, or an anti-platelet agent such as
aspirin, piroxicam or ticlopidine, or a thrombin
inhibitor such as a boropeptide, hirudin or argatroban,
or a thrombolytic agent such as tissue plasminogen
activator, anistreplase, urokinase or streptokinase, or
combinations thereof, can be in any dosage form, such as
those described above, and can also be administered in
various ways, as described above.
in a preferred embodiment, the combination products
of the invention are formulated together, in a single
dosage form (that is, combined together in one capsule,
tablet, powder, or liquid, etc.). When the combination
products are not formulated together in a single dosage
form, the avp3 antagonist compounds of this invention and
the anti-coagulant agent, anti-platelet agent, thrombin
inhibitor, and/or thrombolytic agent may be administered
at the same time (that is, together), or in any order,
for example the compounds of this invention are
administered first, followed by administration of the
anti-coagulant agent, anti-platelet agent, thrombin
inhibitor, and/or thrombolytic agent. When not
administered at the same time, preferably the
administration of the compound of this invention and any
anti-coagulant agent, anti-platelet agent, thrombin
inhibitor, and/or thrombolytic agent occurs less than
about one hour apart, more preferably less than about 30
minutes apart, even more preferably less than about 15
minutes apart, and most preferably less than about 5
minutes apart. Preferably, administration of the
combination products of the invention is oral. The -270-
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terms oral agent, oral inhibitor, oral compound, or the
like, as used herein, denote compounds which may be
orally administered. Although it is preferable that the
a,,03 antagonist compounds of this invention and the
ar-ti-coagulant agent, anti-platelet agent, thrombin
inhibitor, and/or thrombolytic agent are both
administered in,the same fashion (that is, for example,
both orally), if desired, they may each be administered
in different fashions (that is, for example, one
component of the combination product may be administered
orally, and another component may be administered
intravenously). The dosage of the combination products
of the invention may vary depending upon various factors
such as the pharmacodynamic characteristics of the
particular agent and its mode and route of
administration, the age, health and weight of the
recipient, the nature and extent of the symptoms, the
kind of concurrent treatment, the frequency of
treatment, and the effect desired, as described above.
As discussed above, where two or more of the
foregoing therapeutic agents are combined or
co-administered with the compounds of this invention,
generally the amount of each component in a typical
daily dosage and typical dosage form may be reduced
relative to the usual dosage of the agent when
administered alone, in view of the additive or
synergistic effect which would be obtained as a result
of addition of further agents in accordance with the
present invention.
Particularly when provided as a single dosage form,
the potential exists for a chemical interaction between
the combined active ingredients (for example, a novel
compound of this invention and an anti-coagulant such as
warfarin or heparin, or a novel compound of this
invention and an anti-platelet agent such as aspirin,
piroxicam or ticlopidine, or a novel compound of this
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invention and a thrombin inhibitor such as a
boropeptide, hirudin or argatroban, or a novel compound
of this invention and a thrombolytic agent such as
tissue plasminogen activator, anistreplase, urokinase or
streptokinase, or combinations thereof). For this
reason, the preferred dosage forms of the combination
products of this invention are formulated such that
although the active ingredients are combined in a single
dosage form, the physical contact between the active
ingredients is minimized (that is, reduced).
In order to minimize contact, one embodiment of
this invention where the product is orally administered
provides for a combination product wherein one active
ingredient is enteric coated. By enteric coating one of
the active ingredients, it is possible not only to
minimize the contact between the combined active
ingredients, but also, it is possible to control the
release of one of these components in the
gastrointestinal tract such that one of these components
is not released in the stomach but rather is released in
the intestines. Another embodiment of this invention
where oral administration is desired provides for a
combination product wherein one of the active
ingredients is coated with a sustained-release material
which effects a sustained-release throughout the
gastrointestinal tract and also serves to minimize
physical contact between the combined active
ingredients. Furthermore, the sustained-released
component can be additionally enteric coated such that
the release of this component occurs only in the
intestine. Still another approach would involve the
formulation of a combination product in which the one
component is coated with a sustained and/or enteric
release polymer, and the other component is also coated
with a polymer such as a low viscosity grade of
hydroxypropyl methylcellulose (HPMC) or other
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appropriate materials as known in the art, in order to
further separate the active components. The polymer
coating serves to form an additional barrier to
interaction with the other component.
Dosage forms of the combination products of the
present invention wherein one active ingredient is
enteric coated.can be in the form of tablets such that
the enteric coated component and the other active
ingredient are blended together and then compressed into
a tablet or such that the enteric coated component is
compressed into one tablet layer and the other active
ingredient is compressed into an additional layer.
Optionally, in order to further separate the two layers,
one or more placebo layers may be present such that the
placebo layer is between the layers of active
ingredients. In addition, dosage forms of the present
invention can be in the form of capsules wherein one
active ingredient is compressed into a tablet or in the
form of a plurality of microtablets, particles, granules
or non-perils, which are then enteric coated. These
enteric coated microtablets, particles, granules or non-
perils are then placed into a capsule or compressed into
a capsule along with a granulation of the other active
ingredient.
These as well as other ways of minimizing contact
between the components of combination products of the
present invention, whether administered in a single
dosage form or administered in separate forms but at the
same time by the same manner, will be readily apparent
to those skilled in the art, once armed with the present
disclosure.
Pharmaceutical kits useful in, for example, the
inhibition of thrombus formation, the prevention of
blood clots, and/or the treatment of thromboembolic
disorders, which comprise a therapeutically effective
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amount of a compound according to the method of the
present invention along with a therapeutically effective
amount of an anti-coagulant agent such as warfarin or
heparin, or an antiplatelet agent such as aspirin,
piroxicam or ticlopidine, or a thrombin inhibitor such
as a boropeptide, hirudin or argatroban, or a thrombolytic agent such as
tissue plasminogen activator,
anistreplase, urokinase or streptokinase, or
combinations thereof, in one or more sterile containers,
are also within the ambit of the present invention.
Sterilization of the container may be carried out using
conventional sterilization methodology well known to
those skilled in the art. The sterile containers of
materials may comprise separate containers, or one or
more multi-part containers, as exemplified by the
UNIVIALT"t two-part container (available from Abbott Labs,
Chicago, Illinois), as desired. The compounds according
to the method of the invention and the anti-coagulant
agent, anti-platelet agent, thrombin inhibitor,
thrombolytic agent, and/or combinations thereof, may be
separate, or combined into a single dosage form as
described above. Such kits may further include, if
desired, one or more of various conventional
pharmaceutical kit components, such as for example, one
or more pharmaceutically acceptable carriers, additional
vials for mixing the components, etc., as will be
readily apparent to those skilled in the art.
Instructions, either as inserts or as labels, indicating
quantities of the components to be administered,
guidelines for administration, and/or guidelines for
mixing the components, may also be included in the kit.
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