Note: Descriptions are shown in the official language in which they were submitted.
CA 02361285 2008-10-23
PROPANOIC ACID DERIVATIVES THAT INHIBIT THE BINDING OF
INTEGRINS TO THEIR RECEPTORS
Field of the Invention
This invention is directed generally to the inhibition of the binding of a4(3,
integrin to its receptors, for example VCAM-1 (vascular cell adhesion molecule-
1) and fibronectin. The invention also relates to compounds that inhibit this
binding, to pharmaceutically active compositions comprising such compounds
and to the use of such compounds either as above, or in formulations for the
control or prevention of disease states in which a,(3, is involved.
Background of the Invention
When a tissue has been invaded by a microorganism or has been damaged,
white blood cells, also called leukocytes, play a major role in the
inflammatory
response. One of the most important aspects of the inflammatory response
involves the cell adhesion event. Generally, white blood cells are found
circulating through the bloodstream. However, when a tissue is infected or
becomes damaged, the white blood cells recognize the invaded or damaged
tissue, bind to the wall of the capillary and migrate through the capillary
into the
affected tissue. These events are mediated by a family of proteins called cell
adhesion molecules.
There are three main types of white blood cells: granulocytes, monocytes
and lymphocytes. The integrin 4 , (also called VLA-4 for very late antigen-4)
is a heterodimeric protein expressed on the surface of monocytes, lymphocytes
and two subclasses of granulocytes: eosinophils and basophils. This protein
plays a key role in cell adhesion through its ability to recognize and bind
VCAM-1 and fibronectin, proteins associated with thc cndothelial cells that
line
the interior wall of capillaries.
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Following infection or damage of tissue surrounding a capillary, endothelial
cells express a series of adhesion molecules, including VCAM-l, that are
critical for binding the white blood cells that are necessary for fighting
infection.
Prior to binding to VCAM-1 or fibronectin, the white blood cells initially
bind
to certain adhesion molecules to slow their flow and allow the cells to "roll"
along the activated endothelium. Monocytes, lymphocytes, basophils and
eosinophils are then able to firmly bind to VCAM-1 or fibronectin on the blood
vessel wall via the a4P , integrin. There is evidence that such interactions
are
also involved in transmigration of these white blood cells into the damaged
tissue, as well as the initial rolling event itself.
Although white blood cell migration to the site of injury helps fight
~ infection and destroy foreign material, in many instances this migration can
become uncontrolled, with white blood cells flooding to the scene, causing
widespread tissue damage. Compounds capable of blocking this process,
therefore, may be beneficial as therapeutic agents. Thus, it would be useful
to
develop inhibitors that would prevent the binding of white blood cells to
VCAM-1 and fibronectin.
Some of the diseases that might be treated by the inhibition of a4P, binding
include, but are not limited to, atherosclerosis, rheumatoid arthritis,
asthma,
allergy, multiple sclerosis, lupus, inflammatory bowel disease, graft
rejection,
contact hypersensitivity, and type I diabetes. In addition to being found on
some
white blood cells, 41 is also found on various cancer cells, including
leukemia, melanoma, lymphoma and sarcoma cells. It has been suggested that
cell adhesion involving 4 , may be involved in the metastasis of certain
cancers. Inhibitors of aA binding may, therefore, also be useful in the
treatment of some forms of cancer.
The isolation and purification of a peptide which inhibits the binding of
aA to a protein is disclosed in U.S. Patent No. 5,510,332. Peptides which
inhibit binding are disclosed in WO 95/15973, EP 0 341 915, EP 0 422 938 Al,
U.S. Patent No. 5,192,746 and WO 96/06108. Novel compounds which are
useful for inhibition and prevention of cell adhesion and cell adhesion-
mediated
pathologies are disclosed in WO 96/22966, WO 98/04247 and WO 98/04913.
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3
It is therefore an object of the invention to provide novel compounds which
are inhibitors of aaP, binding, and pharmaceutical compositions including such
novel compounds.
Brief SummM of the Invention
The invention is directed to novel compounds of Formula I as follows:
Formula I
X
J M
q C N T/LR4
~ R1 Q
R~ \ Ra
I
wherein circle Q represents one or more rings;
q is an integer of zero to six;
M is selected from the group consisting of -C(R9)(R10)- and
-(CH,),,-, wherein u is an integer of from 0 to 3;
J is selected from the group consisting of -0-, -S- and -NR12-;
T is selected from the group consisting of -C(O)- and -(CHZ)b wherein b is
an integer of from 0 to 3;
~ 15 L is selected from the group consisting of -0-, -NR13-, -S-, and
-(CHZ),.- wherein v is an integer of 0 or 1;
X is selected from the group consisting of -COzB, -P03H2,
-SO3H, -SO2NH2, -SO2NHCOR 14, -OP03HZ, -C(O)NHC(O)R15,
-C(O)NHSO2R16, tetrazolyl, oxazolyl and hydroxyl;
R", R1z and R" are each independently selected from the group consisting
of hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkyl, aliphatic acyl,
alkynylamino, alkoxycarbonyl, heterocycloyl, -CH=NOH, haloalkyl,
alkoxyalkyl, carboxaldehyde, carboxamide, cycloalkyl,
cycloalkenyl, cycloalkynyl, cycloalkylalkyl, aryl, aroyl, aryloxy,
arylamino, biaryl, thioaryl, diarylamino, heterocyclyl, alkylaryl,
aralkenyl, aralkyl, alkylheterocyclyl, heterocyclylalkyl, carbamate,
aryloxyalkyl and -C(O)NH(benzyl) groups; and,
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B, R', R4, R6, R8, R9, R10 , R", R'S and R'6 at each occurrence are
independently selected from the group consisting of hydrogen,
halogen, alkyl, alkenyl, alkynyl, alkoxy, alkenoxy, alkynoxy,
thioalkoxy, hydroxyalkyl, aliphatic acyl,
-CF31 -CO2H, -SH, -CN, -NOZ, -NHZ, -OH, alkynylamino,
alkoxycarbonyl, heterocycloyl, carboxy, -N(CI-C3 alkyl)-C(O)(C,-
C3 alkyl), -NHC(O)N(C,-C3 alkyl)C(O)-NH(C,-C3 alkyl),
-NHC(O)NH(C,-C6alkyl), -NHSO2(C,-C3 alkyl),
-NHSO2(aryl), alkoxyalkyl, alkylamino, alkenylamino, di(C,-
C3)amino, -C(O)O-(C,-C3)alkyl, -C(O)NH-(C,-C3)alkyl,
-C(O)N(C,-C3 alkyl)z, -CH=NOH, -PO3H2, -OP03H2, haloalkyl,
alkoxyalkoxy, carboxaldehyde, carboxamide, cycloalkyl,
cycloalkenyl, cycloalkynyl, cycloalkylalkyl, aryl, aroyl, aryloxy,
arylamino, biaryl, thioaryl, diarylamino, heterocyclyl, alkylaryl,
aralkenyl, aralkyl, alkylheterocyclyl, heterocyclylalkyl, sulfonyl,
-SO,-(C,-C, alkyl), -SO,-(C,-C3 alkyl), sulfonamido, carbamate,
aryloxyalkyl and -C(O)NH(benzyl) groups;
wherein B,R', Ra, R6, Rs, R9, R10 , R", R'z, R'3, R'4 , R15 and R`6are
unsubstituted or substituted with at least one electron
donating or electron withdrawing group;
wherein when L is -NR13-, Ra and R13 taken together may
form a ring;
and wherein R6 and Rg taken together may form a ring;
and wherein when M is -C(R9)(R10)-, R9 and R'0 taken
together may form a ring;
or a pharmaceutically acceptable salt thereof.
For Formula I, presently preferred compounds may have circle Q as an
aryl, cycloalkyl, biaryl or heterocyclyl ring.
More specifically, the compounds of this invention may be
described by Formula II below
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0
R9
O OB
i) Rio
m
L
W--~' C N T/ R4
ill
R6' Ra
O
Formula II
wherein Y, at each occurrence, is independently selected from the group
consisting of C(O), N, CR7, C(RZ)(R3), NRS, CH, 0 and S;
5 misan integeroffrom2to5;
W is selected from the group consisting of C, N and CRZ2;
T is selected from the group consisting of C(O) and (CH,)b wherein b is
an integer of 0 to 3;
L is selected from the group consisting of 0, NR13, S, and
(CH,)n wherein n is an integer of 0 or 1;
R', R" and R" at each occurrence, are each independently selected from
the group consisting of hydrogen, alkyl, alkenyl, alkynyl,
hydroxyalkyl, aliphatic acyl, alkynylamino, alkoxycarbonyl,
heterocycloyl, -CH=NOH, haloalkyl, alkoxyalkyl,
carboxaldehyde, carboxamide, cycloalkyl, cycloalkenyl,
cycloalkynyl, cycloalkylalkyl, aryl, aroyl, aryloxy, arylamino,
biaryl, thioaryl, diarylamino, heterocyclyl, alkylaryl, aralkenyl,
aralkyl, alkylheterocyclyl, heterocyclylalkyl, carbamate,
aryloxyalkyl and -C(O)NH(benzyl) groups; and,
B, R2, R', R~, R6, R', Rg, R9, R10 and R22 are each independently
selected from the group consisting of hydrogen, halogen, alkyl,
alkenyl, alkynyl, alkoxy, alkenoxy, alkynoxy, thioalkoxy,
hydroxyalkyl, aliphatic acyl, -CF3, -COzH, -SH, -CN, -NOz,
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6
-NH2, -OH, alkynylamino, alkoxycarbonyl, heterocycloyl,
carboxy, -N(C,-C3 alkyl)-C(O)(C,-C3 alkyl), -NHC(O)N(C1-C3
alkyl)C(O)-NH(C,-C3 alkyl), -NHC(O)NH(C,-C6 alkyl),
-NHSO2(C,-C3 alkyl), -NHSOz(aryl), alkoxyalkyl, alkylamino,
alkenylamino, di(C,-C3)amino, -C(O)O-(C,-C3)alkyl, -C(O)NH-
(C,-C3)alkyl, -C(O)N(C,-C3 alkyl)2, -CH=NOH, -P03H21
-OP03HZ, haloalkyl, alkoxyalkoxy, carboxaldehyde,
carboxamide, cycloalkyl, cycloalkenyl, cycloalkynyl,
cycloalkylalkyl, aryl, aroyl, aryloxy, arylamino, biaryl, thioaryl,
diarylamino, heterocyclyl, alkylaryl, aralkenyl, aralkyl,
alkylheterocyclyl, heterocyclylalkyl, sulfonyl, -SOz-(C,-C3 alkyl),
-S03-(C,-C3 alkyl), sulfonamido, carbamate, aryloxyalkyl and
-C(O)NH(benzyl) groups;
wherein B, RZ, R3, R4, R5, R6, R7, Ra, R9, R10 , R", R`3 and R22 are
unsubstituted or substituted with at least one electron donating or
electron withdrawing group;
and wherein when L is -NR13-, R4 and R13 taken together may form a
ring; and wherein R6 and R8 taken together may form a ring;
and wherein R9 and R10 taken together may form a ring;
or a pharmaceutically acceptable salt thereof.
~ More specifically, the compounds of this invention may be described by
Formula III below
0
R9
O OB
o
q(R')
N L
C N TR4
~ 8
R6 I 11
Formula III
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wherein q is an integer of zero to four;
T is selected from the group consisting of C(O) and (CHZ)b wherein b is
an integer of 0 to 3;
L is selected from the group consisting of 0, NR13, S, and
(CH2)n wherein n is an integer of 0 or 1;
R" and Rt3 are each independently selected from the group consisting of
hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkyl, aliphatic acyl,
alkynylamino, alkoxycarbonyl, heterocycloyl, -CH=NOH,
haloalkyl, alkoxyalkyl, carboxaldehyde, carboxamide, cycloalkyl,
cycloalkenyl, cycloalkynyl, cycloalkvlalkyl, aryl, aroyl, aryloxy,
arylamino, biaryl, thioaryl, diarylamino, heterocyclyl, alkylaryl,
aralkenyl, aralkyl, alkylheterocyclyl, heterocyclylalkyl,
carbamate, aryloxyalkyl and -C(O)NH(benzyl) groups; and,
B, R', R4, R6, Rg, R9 and R10 at each occurrence are each independently
selected from the group consisting of hydrogen, halogen, alkyl,
alkenvl, alkynyl, alkoxy, alkenoxy, alkynoxy, thi.oalkoxy,
hydroxyalkoxy, aliphatic acyl, -CF3,-CO2H, -SH,
-CN, -NO,, -NH2, -OH, alkynylamino, alkoxycarbonyl,
heterocycloyl, carboxy, -N(C1 -C3 alkyl)-C(O)(C,-C3 alkyl),
-NHC(O)N(C1-C3 alkyl)C(O)NH(C,-C3 alkyl), -NHC(O)NH(C,-
~ C6 alkyl), -NHSO,(C1-C3 alkyl), -NHSO,(aryl), alkoxyalkyl,
alkylamino, alkenylamino, di(C,-C3)amino, -C(O)O-(C,-C3)alkyl,
-C(O)NH(C,-C3)alkyl, -C(O)N(Ci-C3 alkyl)2, -CH=NOH,
-P03HZ, -OPO3H2, haloalkyl, alkoxyalkoxy, carboxyaldehyde,
carboxamide, cycloalkyl, cycloalkenyl, cycloalkynyl,
cycloalkylalkyl, aryl, aroyl, aryloxy, arylamino, biaryl, thioaryl,
diarylamino, heterocyclyl, alkylaryl, araikenyl, aralkyl,
alkylheterocyclyl, heterocyclylalkyl, sulfonyl, -S02-(C1-C3 alkyl),
-S03-(C1-C3 alkyl), sulfonamido, carbamate, aryloxyalkyl and -
C(O)NH(benzyl) groups;
wherein B, R', R;, R6, Rg, R9, R10 , R" and R'' are unsubstituted or
substituted with at least one electron donating or electron
withdrawing group;
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and wherein when L is -NR13-, R4 and R13 taken together may form a
ring;
and wherein R6 and R8 taken together may form a ring;
and wherein R9 and R10 taken together may form a ring;
or a pharmaceutically acceptable salt thereof.
Presently preferred compounds of Formula III have q as one or two; R'
at each occurrence independently as aralkyl or alkyl; R6 as alkyl; B, R8, R9,
R'o
and R" ach independently as hydrogen, T as -(CHz)b wherein b is zero; L as
-(CH,)n- wherein n is zero and R as aryl.
More specifically, the compounds of this invention may be described by
Formula IV below
{
R19 0
R9
R1a Y Rzo
O OB
s(R17 Rio
N N R2~
8
R6 R RII
O
~. .
Formula IV
wherein s is an integer of zero to five;
R'-' is selected from the group consisting of aryl, alkyheterocyclyl,
heterocyclylalkyl, heterocycloyl, aralkyl, alkylaryl, alkyl, aroyl,
aryloxy, cycloalkyl, cycloalkenyl, cycloalkynyl, biaryl,
arylamino, thioaryl and diarylamino;
B, R6, R8, R9, R10, R", R'g, R'9 and R20 at each occurrence are each
independently selected from the group consisting of hydrogen,
halogen, alkyl, alkenyl, alkynyl, alkoxy, alkenoxy, alkynoxy,
thioalkoxy, hydroxyalkoxy, aliphatic acyl, -CF3, -COzH, -SH,
-CN, NO,, -NH,, -OH, alkynylamino, alkoxycarbonyl,
heterocycloyl, carboxy, -N(C,-C3 alkyl)-C(O)(C,-C3 alkyl),
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-NHC(O)N(C,-C3 alkyl)C(O)NH(C,-C3 alkyl), -NHC(O)NH(C,-
C6 alkyl), -NHSO2(C1-C3 alkyl), -NHS02(aryl), alkoxyalkyl,
alkylamino, alkenylamino, di(C,-C3)amino, -C(O)O-(C,-C3)alkyl,
-C(O)NH(C1-C3)alkyl, -C(O)N(C1-C3 alkyl)Z, -CH=NOH,
-P03H21 -OP03HZ, haloalkyl, alkoxyalkoxy, carboxyaldehyde,
carboxamide, cycloalkyl, cycloalkenyl, cycloalkynyl,
cycloalkylalkyl, aryl, aroyl, aryloxy, arylamino, biaryl, thioaryl,
diarylamino, heterocyclyl, alkylaryl, aralkenyl, aralkyl,
alkylheterocyclyl, heterocyclylalkyl, sulfonyl, -S02-(C,-C3 alkyl),
-S03-(C,-C3 alkyl), sulfonamido, carbamate, aryloxyalkyl and -
C(O)NH(benzyl) groups; and,
_.-,-
R" is selected from the group consisting of hydrogen, alkyl, alkenyl,
alkynyl, hydroxyalkyl, aliphatic acyl, alkynylamino,
alkoxycarbonyl, heterocycloyl, -CH=NOH, haloalkyl,
alkoxyalkyl, carboxaldehyde, carboxamide, cycloalkyl,
cycloalkenyl, cycloalkynyl, cycloalkylalkyl, aryl, aroyl, aryloxy,
arylamino, biaryl, thioaryl, diarylamino, heterocyclyl, alkylaryl,
aralkenyl; aralkyl, alkylheterocyclyl, heterocyclylalkyl,
carbamate, aryloxyalkyl and -C(O)NH(benzyl) groups;
wherein B, R6, Rg, R9, R10, R", R", R18, R'9, R20 and R21 are
unsubstituted or substituted with at least one electron donating or
electron withdrawing group;
and wherein R6 and Rg taken together may form a ring;
and wherein R9 and R10 taken together may form a ring;
and wherein R'8 and R19 taken together may form a ring;
and wherein R'9 and R20 taken together may form a ring;
or a pharmaceutically acceptable salt thereof.
Presently preferred compounds of Formula IV have B, Rg, R9, R10, R",
R'g and R20 each independently as hydrogen, R6 and R19 each independently as
alkyl; s as an integer of zero to three; R" at each occurrence independently
as
halogen, alkyl, haloalkyl, -CF3, alkoxy or -OH; and RZ' as aryl. A presently
most preferred compound of Forrnula IV has s as zero; R6 as butyl; B, Rg, R9,
CA 02361285 2008-10-23
R10, R", R'$ and R20 each independently as hydrogen; and RZ' as 1,3-
benzodioxol-5-yl.
In another embodiment of the present invention, there is provided a
compound of the structure:
5
0
R9
\R~ ~9
( O K N ~o
~ OR
O
R 6 R8 I I
O R>>
O
or a pharmaceutically acceptable salt thereof,
wherein
q is an integer from 0 to 4;
B, R', R9, R1 and R" are each independently hydrogen;
R' at each occurrence is independently selected from the group
consisting of C i _3alkyl, aryl(C 1_3alkyl), (C I _3alkyloxy)aryl(C 1
_3alkyl), (C i _
3alkyl)aryl(Ci_3alkyl), haloaryl(Ci_3alkyl), thioaryl and arylcarbonyl; and
R 8 at each occurrence is independently selected from the group consisting
of (Ci-C6)alkanoyl and (C3-C6)cycloalkyl.
Compounds of the present invention may also be described by Formula V,
shown below.
0
R9
O OB
R10
R Q C N R21
q
R6 R i
Formula V
CA 02361285 2008-10-23
10a
wherein circle Q is a ring selected from the group consisting of
q is an integer of zero to four;
\R~/q~ q ~ N
N IN"~ ~~
Rs// ~ 1-
0 and 0
B, RR6, R8, R4 and R10 are each independently selected from the group
consisting of hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy,
alkenoxy, alkynoxy, thioalkoxy, hydroxyalkyl, aliphatic acyl,
-CF3, -COZH, -SH, -CN, NOZ, -OH, alkynylamino,
alkoxycarbonyl, heterocycloyl, carboxy, -N(CI -C3 alkyl)-
C(O)( Cz-C3 alkyl), -NHC(O)NH(Ci-C3 alkyl) -NHC(O)N(CI-C3 alkyl)C(O)NH(Ci-C3
alkyl), -NHSO,)(Ci-C3 alkyl),
-NHSO2(aryl), alkoxyalkyl,-Cl-C3 alkylamino, alkenylamino,
alkynylamino, di(C I -C3 alkyl)amino, -C(O)O-(C I -C3 alkyl),
-C(O)NH-(Ci-C3 alkyl), -CH=NOH, -P03H2, -OP03H2,
-C(O)N(Ci-C3 alkyl)2, haloalkyl, alkoxyalkoxy, carboxaldehyde,
carboxamide, cycloalkyl, cycloalkenyl, cycloalkynyl,
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cycloalkylalkyl, aryl, aroyl, aryloxy, arflamino, biaryl, thioaryl,
diarylamino, heterocyclyl, alkylaryl, aralkenyl, aralkyl,
alkylheterocyclyl, heterocyclylalkyl, sulfonyl, -SO2-(C,-C3 alkyl),
-S03-(C1-C3 alkyl), sulfonamido, aryloxyalkyl, carboxyl,
carbamate and -C(O)NH(benzyl);
RS and R" are each independently selected from the group consisting of
hydrogen, alkyl, alkenyl, alkynyl, hydroxyalkyl, aliphatic acyl,
alkynylamino, alkoxycarbonyl, heterocycloyl, -CH=NOH,
haloalkyl, alkoxyalkyl, carboxaldehyde, carboxamide, cycloalkyl,
cycloalkenyl, cycloalkynyl, cycloalkylalkyl, aryl, aroyl, aryloxy,
arylamino, biaryl, thioaryl, diarylamino, heterocyclyl, alkylaryl,
aralkenyl, aralkyl, alkylheterocyclyl, heterocyclylalkyl,
carbamate, aryloxyalkyl and -C(O)NH(benzyl) groups; and,
RZ` is selected from the group consisting of aryl, alkyheterocyclyl,
heterocyclylalkyl, heterocycloyl, aralkyl, alkylaryl, alkyl, aroyl,
aryloxy, cycloalkyl, cycloalkenyl, cycloalkynyl, biaryl,
arylamino, thioaryl and diarylamino;
wherein B, R', R5, R6, R8, R9, R10, R" and R" are unsubstituted or
substituted with at least one electron donating or electron
withdrawing group;
and wherein R6 and R8 taken together may form a ring;
and wherein R9 and R10 taken together may form a ring;
or a pharmaceutically acceptable salt thereof.
Presently preferred compounds of Formula V have B, R6, Rg, R9 , R10
and R." each independently as hydrogen and alkyl, R' and R5, at each
occurrence, independently as hydrogen, 2-thienylmethyl, benzyl or methyl and
R21 as aryl.
Presently preferred compounds include (3S)-(1,3-benzodioxol-5-yl)-3-
(((2S)-2-(2-oxo-3-(2-thienylmethyl)tetrahydro-1(2H)-
pyrimidinyl)hexanoyl)amino) propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-
((2R, S)-2-(3-benzyl-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic
acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-(3-chlorobenzyl)-5-methyl-2-
oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid, (3S)-3-(1,3-benzodioxol-
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5-yl)-3-(((2S)-2-(2-oxo-3-(phenylmethyl)-1(2H)-pyridinyl)hexanoyl)amino)
propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2-(3-chlorophenyl)methyl)-5-
methyl-2-oxo-1(2H)-pyridinyl)hexanoyl)amino)propanoic acid, (3S)-3- {[(2S)-2-
(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl] amino } -3 -(3,5-
dimethoxyphenyl)propanoic acid, (3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2-
oxopyridin-1(2H)-yl)hexanoyl]amino} -3 -(3 -fluoro-4-methoxyphenyl)propanoic
acid, (3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino}-3-
(3-isopropoxyphenyl)propanoic acid, (3S)-3-( {2-[3-(3-chlorobenzyl)-5-methyl-
2-oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(3-isopropoxyphenyi)propanoic
acid, (3S)-3-(3-isopropoxyphenyl)-3-( {2-[3-(2-methoxybenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl}amino)propanoic acid, (3S)-3-{[2-(3-benzyl-5-
methY1-2-oxoPYridin-1 (2H)-Y1)hexanoY1] amino } -3-(3-ethoxy-4-
methoxyphenyl)propanoic acid, (3 S)-3- { [2-(3-benzyl-5-methyl-2-oxopyridin-
1(2H)-yl)hexanoyl]amino1 -3-(4-methoxy-3-methylphenyl)propanoic acid, (3S)-
3- { [2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino } -3-
phenylpropanoic acid, (3S)-3-( {2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-
1(2H)-yl]hexanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-3-({2-[3-(2-
chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(3-
isopropoxyphenyl)propanoic acid, (3S)-3-( {2-[3-(2-chlorobenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl}amino)-3-phenylpropanoic acid, (3S)-3-(1-
methyl-1 H-indol-6-yl)-3-( {2-[5-methyl-2-oxo-3-(phenylmethyl)-1(2H)-
pyridinyl]hexanoyl}amino)propanoic acid, (3S)-3-{3-[(1-
methylethyl)oxy] phenyl } -3-( { 2-[ 5-methyl-3 -(2-naphthalenylmethyl)-2-oxo-
1(2H)-pyridinyl]hexanoyl} amino)propanoic acid, (3S)-3-( {2-[5-methyl-2-oxo-
3-(phenylmethyl)-1(2H)-pyridinyl]hexanoyl} amino)-3-[3-(2-
methylpropyl)phenyl]propanoic acid, (3S)-3-[3-(difluoromethyl)phenyl]-3-( {2-
[5-methyl-2-oxo-3-(phenylmethyl)-1(2H)-pyridinyl]hexanoyl} amino)propanoic
acid, (3S)-3- {3-[(1-methylethyl)oxy]phenyl}-3-( {2-[5-methyl-3-[(2-
methylphenyl)methyl]-2-oxo-1(2H)-pyridinyl]hexanoyl}amino)propanoic acid,
(3S)-3-(3-fluorophenyl)-3-({2-[5-methyl-2-oxo-3-(phenylmethyl)-1(2H)-
pyridinyl]hexanoyl} amino)propanoic acid, (3S)-3-( {2-[3-[(2-chloro-4-
fluorophenyl)methyl]-5-methyl-2-oxo-1(2H)-pyridinyl]hexanoyl} amino)-3- {3-
[(1-methylethyl)oxy]phenyl}propanoic acid and (3S)-3-{3-[(1-
CA 02361285 2001-11-07
13
methylethyl)oxy]phenyl}-3-( {2-[5-methyl-2-oxo-1-(phenylmethyl)-1,2-dihydro-
3-pyridinyl]-2-phenylacetyl}amino) propanoic acid, and pharmaceutically
acceptable salts thereof.
Other presently preferred compounds include
(3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino}-3-
(3',4'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-
chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(3',4'-
dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-3-(1,1'-biphenyl-4-yl)-3-
( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)propanoic acid, (3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2-
oxopyridin-1(2H)-yl)hexanoyl] amino) -3-(1,1'-biphenyl-4-yl)propanoic acid,
(3 S)-3-(1,1'-biphenyl-4-yl)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl}amino)propanoic acid, (3S)-3-{[(2S)-2-(3-
benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino} -3-(4'-methyl-1,1'-
biphenyl-4-yl)propanoic acid, (3S)-3-({(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl}amino)-3-(4'-methyl- 1,1'-biphenyl-4-
yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl] hexanoyl } amino)-3-(4'-methyl-1,1'-biphenyl-4-
yl)propanoic acid, (3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-
yl)hexanoyl]amino}-3-(2',6'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-
~: _;.
yl]hexanoyl} amino)-3-(2',6'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3-( { (2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)-3-(2',6'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3-{[(2S)-2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino}-3-(1,1'-
biphenyl-3-yl)propanoic acid, (3S)-3-(1,1'-biphenyl-3-yl)-3-({(2S)-2-[3-(2-
chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyi} amino)propanoic acid,
(3S)-3-(1,1'-biphenyl-3-yl)-3-( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-
2-oxopyridin-1(2H)-ylJhexanoyl}amino)propanoic acid, (3S)-3-{[(2S)-2-(3-
benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino}-3-(2'-methoxy-1,1'-
biphenyl-3-yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(2'-methoxy-1,1'-biphenyl-3-
yl)propanoic acid, (3S)-3-({(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-
CA 02361285 2001-11-07
14
oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(2'-methoxy-1,1'-biphenyl-3-
yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-fluoro-6-methoxybenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-
3-( {(2S)-2-[3-(2,6-dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-3-({(2S)-2-[3-(3,5-
dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(4-
methylphenyl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2,6-dichlorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(4-methylphenyl)propanoic
acid, (3S)-3-( {(2S)-2-[3-(2-chloro-6-fluorobenzyl)-5-methyl-2-oxopyridin-
1(2H)-yl]hexanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-3-({(2S)-2-
[3-(2,6-difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(4-
.:-
methylphenyl)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[3-(2,6-
difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)propanoic
acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[3-(2-chloro-6-fluorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl}amino)propanoic acid, (3S)-3-(1,3-
benzodioxol-5-yl)-3-( {(2S)-2-[3-(2,6-dichlorobenzyl)-5-methyl-2-oxopyridin-
1(2H)-yl]hexanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-
( {(2S)-2-[3-(2,6-dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-
[3-(3,5-dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-
~.
[3-(2-fluoro-6-methoxybenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl} amino)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(3,4-
diethoxyphenyl)propanoic acid, (3S)-3-(3,4-diethoxyphenyl)-3-({(2S)-2-[3-(2,6-
difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)propanoic
acid, (3S)-3-( {(2S)-2-[3-(2,6-difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl} amino)- 3 -(3 -ethoxyphenyl)propanoic acid, (3 S)-3-( {(2S)-2-[3-
(2-
chloro-6-fluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(3-
ethoxyphenyl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]-3-methylbutanoyl } amino)-3-(4-methylphenyl)propanoic
acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]-4-
methylpentanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-3-(1,3;
CA 02361285 2001-11-07
benzodioxol-5-yl)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-
1(2H)-yl]-4-methylpentanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-
yl)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]-3-
methylbutanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-
5 2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]-3-
phenylpropanoyl}amino)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]-3-phenylpropanoyl} amino)-3-(4-
methylphenyl)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-(((2R)-2-(1-
benzyl-5-methyl-2-oxo-1,2-dihydropyridin-3-yl)hexanoyl)amino) propanoic
10 acid and pharmaceutically acceptable salts thereof.
A presently most preferred compound is (3S)-3-(1,3-benzodioxol-5-yl)-
~ 3-((2S)-2-(3-benzyl-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic
acid or a pharmaceutically acceptable salt thereof.
Derivatives of Formulae I-V which are esters, carbamates, aminals,
15 amides, optical isomers and pro-drugs are also contemplated.
The present invention also relates to pharmaceutical compositions
comprising a physiologically acceptable diluent and at least one compound ot
the present invention.
The present invention further relates to a process of inhibiting the
binding of a4(3, integrin to VCAM-1 comprising exposure of a cell expressing
a4p , integrin to a cell expressing VCAM-1 in the presence of an effective
~._
inhibiting amount of a compound of the present invention. The VCAM-1 may
be on the surface of a vascular endothelial cell, an antigen presenting cell,
or
other cell type. The a,P, may be on a white blood cell such as a monocyte,
lymphocyte, granulocyte; a stem cell; or any other cell that naturally
expresses
aaP I =
The invention also provides a method for treating disease states
mediated by aA binding which comprises administration of an effective
amount of a compound of the present invention, either alone or in formulation,
to an afflicted patient.
CA 02361285 2001-11-07
16
Detailed Description of the Invention
Definition of Terms
The term "alkyl" as used herein, alone or in combination, refers to C,-
C,Z straight or branched, substituted or unsubstituted saturated chain
radicals
derived from saturated hydrocarbons by the removal of one hydrogen atom,
unless the term alkyl is preceded by a CX-Cy designation. Representative
examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl,
sec-butyl, iso-butyl, and tert-butyl among others.
The term "alkenyl" as used herein, alone or in combination, refers to a
substituted or unsubstituted straight-chain or substituted or unsubstituted
branched-chain alkenyl radical containing from 2 to 10 carbon atoms. Examples
of such radicals include, but are not limited to, ethenyl, E- and Z-pentenyl,
decenyl and the like.
The term "alkynyl" as used herein, alone or in combination, refers to a
substituted or unsubstituted straight or substituted or unsubstituted branched
chain alkynyl radical containing from 2 to 10 carbon atoms. Examples of such
radicals include, but are not limited to ethynyl, propynyl, propargyl,
butynyl,
hexynyl, decynyl and the like.
The term "lower" modifyin- "alkyl", "alkenyl", "alkynyl" or "alkoxy"
refers to a C,-C6 unit for a particular functionality. For example lower alkyl
~ means C,-C6 alkyl.
The term "aliphatic acyl" as used herein, alone or in combination, refers
to radicals of formula alkyl-C(O)-, alkenyl-C(O)- and alkynyl-C(O)- derived
from an alkane-, alkene- or alkyncarboxylic acid, wherein the terms "alkyl",
"alkenyl" and "alkynyl" are as defined above. Examples of such aliphatic acyl
radicals include, but are not limited to, acetyl, propionyl, butyryl, valeryl,
4-
methylvaleryl, acryloyl, crotyl, propiolyl and methylpropiolyl, among others.
The term "cycloalkyl" as used herein refers to an aliphatic ring system
having 3 to 10 carbon atoms and I to 3 rings, including, but not limited to
cyclopropyl, cyclopentyl, cyclohexyl, norbornyl, and adamantyl among others.
Cycloalkyl groups can be unsubstituted or substituted with one, two or three
substituents independently selected from lower alkyl, haloalkyl, alkoxy,
CA 02361285 2001-11-07
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thioalkoxy, amino, alkylamino, dialkylamino, hydroxy, halo, mercapto, nitro,
carboxaldehyde, carboxy, alkoxycarbonyl and carboxamide.
"Cycloalkyl" includes cis or trans forms. Furthermore, the substituents
may either be in endo or exo positions in the bridged bicyclic systems.
The term "cycloalkenyl" as used herein alone or in combination refers to
a cyclic carbocycle containing from 4 to 8 carbon atoms and one or more double
bonds. Examples of such cycloalkenyl radicals include, but are not limited to,
cyclopentenyl, cyclohexenyl, cyclopentadienyl and the like.
The term "cycloalkylalkyl" as used herein refers to a cycloalkyl group
appended to a lower alkyl radical, including, but not limited to
cyclohexylmethyl.
The term "halo" or "halogen" as used herein refers to I, Br, Cl or F.
The term "haloalkyl" as used herein refers to a lower alkyl radical, to
which is appended at least one halogen substituent, for example chloromethyl,
fluoroethyl, trifluoromethyl and pentafluoroethyl among others.
The term "alkoxy" as used herein, alone or in combination, refers to an
alkyl ether radical, wherein the term "alkyl" is as defined above. Examples of
suitable alkyl ether radicals include, but are not limited to, methoxy,
ethoxy, n-
propoxy, iso-propoxy, n-butoxy, iso-butoxy, sec-butoxy, tert-butoxy and the
like.
~ The term "alkoxyalkyl" as used herein, refers to R},-O-RZ, wherein R}, is
lower alkyl as defined above, and RZ is alkylene (-(CHz),,-) wherein w is an
integer of from one to six.
The term "alkenoxy" as used herein, alone or in combination, refers to a
radical of formula alkenyl-O-, provided that the radical is not an enol ether,
wherein the term "alkenyl" is as defined above. Examples of suitable alkenoxy
radicals include, but are not limited to, allyloxy, E- and Z- 3-methyl-2-
propenoxy and the like.
The term "alkynoxy" as used herein, alone or in combination, refers to a
radical of formula alkynyl-O-, provided that the radical is not an -ynol
ether.
Examples of suitable alkynoxy radicals include, but are not limited to,
propargyloxy, 2-butynyloxy and the like.
CA 02361285 2001-11-07
18
The term "thioalkoxy" refers to a thioether radical of formula alkyl-S-,
wherein "alkyl" is as defined above.
The term "sulfonamido" as used herein refers to -SOZNHZ.
The term "carboxaldehyde" as used herein refers to -C(O)R wherein R is
hydrogen.
The terms "carboxamide" or "amide" as used herein refer to -
C(O)NRaRb wherein Ra and Rb are each independently hydrogen, alkyl or any
other suitable substituent.
The term "carboxy" as used herein refers to -C(O)O-.
The term "alkoxyalkoxy" as used herein refers to RO-RdO- wherein Rc
is lower alkyl as defined above and Rd is alkylene wherein alkylene is -
(CHZ)n'-
,.,.
wherein n' is an integer from 1 to 6. Representative examples of alkoxyalkoxy
groups include methoxymethoxy, ethoxymethoxy, t-butoxymethoxy among
others.
The terrn "alkylamino" as used herein refers to ReNH- wherein Re is a
lower alkyl group, for example, ethylamino, butylamino, among others.
The term "alkenylamino" as used herein, alone or in combination, refers
to a radical of formula alkenyl-Nl-I-or (alkenyl)2N-, wherein the term
"alkenyl"
is as defined above, provided that the radical is not an enamine. An example
of
such alkenylamino radical is the allylamino radical.
The term "alkynylamino" as used herein, alone or in combination, refers
to a radical of formula alkynyl-NH- or (alkynyl)2N- wherein the term "alkynyl"
is as defined above, provided that the radical is not an amine. An example of
such alkynylamino radicals is the propargyl amino radical.
The term "dialkylarr. ino" as used herein refers to RfRN- wherein R f and
Rg are independently selected from lower alkyl, for example diethylamino, and
methyl propylamino, among others.
The term "alkoxycarbonyl" as used herein refers to an alkoxyl group as
previously defined appended to the parent molecular moiety through a carbonyl
group. Examples of alkoxycarbonyl include methoxycarbonyl, ethoxycarbonyl,
and isopropoxycarbonyl among others.
The term "aryl" or "aromatic" as used herein alone or in combination,
refers to a substituted or unsubstituted carbocyclic aromatic group having
about
CA 02361285 2001-11-07
19
6 to 12 carbon atoms such as phenyl, naphthyl, indenyl, indanyl, azulenyl,
fluorenyl and anthracenyl; or a heterocyclic aromatic group which is an
aromatic
ring containing at least one endocyclic N, 0 or S atom such as furyl, thienyl,
pyridyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, 2-pyrazolinyl,
pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl, 1,2,3-triazolyl,
1,3,4-
thiadiazolyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,3,5-triazinyl, 1,3,5-
trithianyl,
indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furanyl, 2,3-
dihydrobenzofuranyl, benzo[b]thiophenyl, 1H-indazolyl, benzimidazolyl,
benzthiazolyl, purinyl, 4H-quinoliziriyl, isoquinolinyl, cinnolinyl,
phthalazinyl,
quinazolinyl, quinoxalinyl, 1,8-naphthridinyl, pteridinyl, carbazolyl,
acridinyl,
phenazinyl, phenothiazinyl, phenoxyazinyl, pyrazolo[1,5-c]triazinyl and the
like. "Aralkyl" and "alkylaryl" employ the term "alkyl" as defined above.
Rings may be multiply substituted.
The term "aralkyl" as used herein, alone or in combination, refers to an
aryl substituted alkyl radical, wherein the terms "alkyl" and "aryl" are as
defined
above. Examples of suitable aralkyl radicals include, but are not limited to,
phenylmethyl, phenethyl, phenylhexyl, diphenylmethyl, pyridylmethyl,
tetrazolyl methyl, furylmethyl, imidazolyl methyl, indolylmethyl,
thienylpropyl
and the like.
The term "aralkenyl" as used herein, alone or in combination, refers to
an aryl substituted alkenyl radical, wherein the terms "aryl" and "alkenyl"
are as
defined above.
The term "arylamino" as used herein, alone or in combination, refers to a
radical of formula aryl-NH-, wherein "aryl" is as defined above. Examples of
arylamino radicals include, but are not limited to, phenylamino(anilido),
naphthlamino, 2-, 3-, and 4- pyridylamino and the like.
The term "benzyl" as used herein, alone or in combination, refers to
C6H5-CH2-.
The term "biaryl" as used herein, alone or in combination, refers to a
radical of formula aryl-aryl, wherein the term "aryl" is as defined above.
The term "thioaryl" as used herein, alone or in combination, refers to a
radical of formula aryl-S-, wherein the term "aryl" is as defined above. An
example of a thioaryl radical is the thiophenyl radical.
CA 02361285 2001-11-07
The term "aroyl" as used herein, alone or in combination, refers to a
radical of formula aryl-CO-, wherein the term "aryl" is as defined above.
Examples of suitable aromatic acyl radicals include, but are not limited to,
benzoyl, 4-halobenzoyl, 4-carboxybenzoyl, naphthoyl, pyridylcarbonyl and the
5 like.
The term "heterocyclyl" as used herein, alone or in combination, refers
to a non-aromatic 3- to 10- membered ring containing at least one endocyclic
N,
0, or S atom. The heterocycle may be optionally aryl-fused. The heterocycle
may also optionally be substituted with at least one substituent which is
10 independently selected from the group consisting of hydrogen, halogen,
hydroxyl, amino, nitro, trifluoromethyl, trifluoromethoxy, alkyl, aralkyl,
alkenyl,
alkynyl, aryl, cyano, carboxy, carboalkoxy, carboxyalkyl, oxo, arylsulfonyl
and
aralkylaminocarbonyl among others.
The term "alkylheterocyclyl" as used herein refers to an alkyl group as
15 previously defined appended to the parent molecular moiety through a
heterocyclyl group, including but not limited to 2-methyl-5-thiazolyl, 2-
methyl-
1-pyrrolyl and 5-ethyl-2-thiophenyl.
The term "heterocyclylalkyl" as used herein refers to a heterocyclyl
group as previously defined appended to the parent molecular moiety through an
20 alkyl group, including but not limited to 2-thienylmethyl, 2-
pyridinylmethyl and
2-(1-piperidinyl) ethyl.
The term "aminal" as used herein refers to a hemi-acetal of the structure
Rt,C(NR;Rt)(NRkR,)- wherein Rh, R, R,, Rk and R, are each independently
hydrogen, alkyl or any other suitable substituent.
The term "ester" as used herein refers to -C(O)Rnõ wherein Rm is
hydrogen, alkyl or any other suitable substituent.
The term "carbamate" as used herein refers to compounds based on
carbamic acid NH,C(O)OH.
The term "optical isomers" as used herein refers to compounds which
differ only in the stereochemistry of at least one atom, including
enantiomers,
diastereomers and racemates.
Use of the above terms is meant to encompass substituted and
unsubstituted moieties. Substitution may be by one or more groups such as
CA 02361285 2008-10-23
21
alcohols, ethers, esters, amides, sulfones, sulfides, hydroxyl, nitro, cyano,
carboxy, amines, heteroatoms, lower alkyl, lower alkoxy, lower
alkoxycarbonyl, alkoxyalkoxy, acyloxy, halogens, trifluoromethoxy,
trifluoromethyl, alkyl, aralkyl, alkenyl, alkynyl, aryl, cyano, carboxy,
carboalkoxy, carboxyalkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl,
alkylheterocyclyl, heterocyclylalkyl, oxo, arylsulfonyl and
aralkylaminocarbonyl
or any of the substituents of the preceding paragraphs or any of those
substituents either attached directly or by suitable linkers. The linkers are
typically short chains of 1-3 atoms containing any combination of -C-, -C(O)-,
-
NH-, -S-, -S(O)-, -0-, -C(O)O- or -S(0)O-. Rings may be substituted multiple
times.
The terms "electron-withdrawing" or "electron-donating" refer to the
ability of a substituent to withdraw or donate electrons relative to that of
hydrogen if hydrogen occupied the same position in the molecule. These terms
are well-understood by one skilled in the art and are discussed in Advanced
Organic Chemistry by J. March, 1985, pp. 16-18. Electron withdrawing groups
include halo, nitro, carboxyl, lower alkenyl, carboxaldehyde, carboxyamido,
aryl,
quatemary ammonium, trifluoromethyl, sulfonyl and aryl Iower alkanoyl among
others. Electron sulfonyl donating groups include such groups as hydroxy,
lower
alkyl, amino, lower alkylamino, di(lower alkyl)amino, aryloxy, mercapto, lower
alkylthio, lower alkylmercapto, and disulfide among others. One skilled in the
art
will appreciate that the aforesaid substituents may have electron donating or
electron withdrawing properties under different chemical conditions. Moreover,
the present invention contemplates any combination of substituents selected
from
the above-identified groups.
The most preferred electron donating or electron withdrawing
substituents are halo, nitro, alkanoyl, carboxaldehyde, arylalkanoyl, aryloxy,
carboxyl, carboxamide, cyano, sulfonyl, sulfoxide, heterocyclyl, guanidine,
quaternary ammonium, lower alkenyl, lower alkynyl, sulfonium salts, hydroxy,
lower alkoxy, lower alkyl, amino, lower alkylamino, di(lower alkyl)amino,
amine lower alkyl mercapto, mercaptoalkyl, alkylthio, alkyldithio, carboxy
lower alkyl, arylalkoxy, alkanoylamino, alkanoyl(lower alkyl)amino, lower
CA 02361285 2001-11-07
22
alkylsufonylamino, arylsulfonylamino, alkylsulfonyl(lower alkyl)amino,
arylsulfonyl(lower alkyl)amino, lower alkylcarboxamide, di(lower
alkyl)carboxamide, sulfonamide, lower alkylsulfonamide, di(lower
alkyl)sulfonamide, lower alkylsulfonyl, and arylsulfonyl.
As used herein, the term "composition" is intended to encompass a
product comprising the specified ingredients in the specified amounts, as well
as
any product which results, directly or indirectly, from a combination of the
specified ingredients in the specified amounts.
The ring including Y in Formula II or ring Q in Formulae I and V can be
a mono-cyclic heterocycle or aromatic ring, or can be a bicyclic ring. When
more than one Y is C(RZ)(R3), the C substituents from each Y may be joined to
~-:
form a ring. Moreover, in the rings including Y, two independent RZ, R3, RS
and R' groups taken together may be linked to form a ring.
Suitable substituents for the aryl, alkyl, cycloalkyl, heterocyclyl groups
or ring including Y defined above, when present, include alcohols, amines,
heteroatoms, or any combination of aryl, alkyl, cycloalkyl or heterocyclyl
groups
either attached directly, or via suitable linkers. The linkers are typically
short
chains of 1-3 atoms containing any combination of C, C=O, C02, 0, N, S, S=O,
SOZ, as for example ethers, amides, amines, ureas, sulfamides, sulfonamides,
and the like.
In the Formulae I and III-V, defined R groups R' and Rt' potentially
substitute their associated rings a number of times. For R`, when q is zero,
the
associated ring is unsubstituted, having hydrogens at each available position.
Similarly, for R", when s is zero, the associated ring is unsubstituted,
having
hydrogens at each available position.
For example, R`, R2, R3 , R5, R', R" and R13 in Formulae I-V above may
independently be, but are not limited to, phenyl, thienylmethyl, isobutyl, n-
butyl,
2-thienylmethyl, 1,3-thiazol-2-yl-methyl, benzyl, thienyl, 3-pyridinylmethyl,
3-
methyl-l-benzothiophen-2-yl, allyl, 3-methoxybenzyl, propyl, 2-ethoxyethyl,
cyclopropylmethyl, benzylsulfanylmethyl, benzylsulfonylmethyl,
phenylsulfanylmethyl, phenethylsulfanylmethyl, 3-phenylpropylsulfanylmethyl,
4-((2-toluidinocarbonyl)amino)benzyl, 2-pyridinylethyl, 2-(1H-indol-3-
yl)ethyl,
1H-benzimidazol-2-yl, 4-piperidinylmethyl, 3-hydroxy-4-methoxybenzyl, 4-
CA 02361285 2001-11-07
23
hydroxyphenethyl, 4-aminobenzyl, phenylsulfonvlmethyl, 4-
(acetylamino)phenyl, 4-methoxyphenyl, 4-aminophenyl, 4-chlorophenyl, (4-
(benzylsulfonyl)amino)phenyl, (4-(methylsulfonyl)amino)phenyl, 2-
aminophenyl, 2-methylphenyl, isopropyl, 2-oxo-l-pyrrolidinyl, 3-
(methylsulfanyl)propyl, (propylsulfanyl)methyl, octylsulfanylmethyl, 3-
aminophenyl, 4-((2-toluidinocarbonyl)amino)phenyl, 2-
((methylbenzyl)amino)benzyl, methylsulfanylethyl, ethylsulfanylmethyl, 2-
chlorobenzyl, 2-bromobenzyl, 2-fluorobenzyl, 2-chloro-6-fluorobenzyl, 2-
chloro-4-fluorobenzyl, 2,4-dichlorobenzyl, 2-chloro-6-methoxybenzyl, 2-
cyanobenzyl, 2,6-difluorobenzyl, 2-chloro-5-(trifluoromethyl)benzyl, 2-chloro-
6-methylbenzyl, 2,6-dimethoxybenzyl, 2-chloro-5-(methylsulfonyl)benzyl, 2-
chloro-6-cyanobenzyl, 2-chloro-6-ethoxybenzyl, 2-chloro-5-methoxybenzyl, 2-
chloro-5-fluorobenzyl, 5-chloro-2-fluorobenzyl, ethyl, propyl, butyl, pentyl,
cyclopropyl, tert-butylamino, propylamino, 4-methyl-l-piperazinyl, 1-
azetidinyl,
4-morpholino, (4-carboxyphenyl)amino, pivaloylamino, ((tert-
butylamino)carbonyl)amino, trifluoromethyl, benzyloxy, 2-(2-
methoxyethoxy)ethoxy, 2-(2-(2-methoxyethoxy)ethoxy)ethoxy and 2-(2-(2-(2-
methoxyethoxy)ethoxy)ethoxy)ethoxy. R6 and Rg may be linked to form a ring
such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 4-piperidinyl, and 4-
tetrahydropyranyl among others. R4 and R" may be linked to form a ring such
as pyrrolidino, 1-piperidino, 4-methyl-l-piperazino, 4-acPto-l-piperazino, and
4-morpholino among others. R9 and R10 may be linked to form a ring such as
cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl among others.
The R4 substituent for Formulae I-111 and the R21 substituent of Formula
IV above may be, but are not limited to, 1,3-benzodioxol-5-yl, 1-naphthyl,
thienyl, 4-isobutoxyphenyl, 2,6-dimethylphenyl, allyloxyphenyl, 3-bromo-4-
methoxyphenyl, 4-butoxyphenyl, 1-benzofuran-2-yl, 2-thienylmethyl, phenyl,
methysulfanyl, phenylsulfanyl, phenethylsulfanyl, 4-bromo-2-thienyl, 3-methyl-
2-thienyl, 4,5-dihydro-1,3-oxazol-2-yl, 3-(trifluoromethyl)phenyl, 4-
(trifluoromethoxy)phenyl, 2,3-dihydro-1,4-benzodioxin-6-yl, 7-methoxy-1,3-
benzodioxol-5-yl, 3-ethoxy-4-methoxyphenyl, 3,4-dimethoxyphenyl, 3,4-
diethoxyphenyl, 3-ethoxyphenyl, 3-methoxy-4-methylphenyl, 3,5-dimethoxy-4-
methylphenyl, 3-propoxyphenyl, 3-butoxyphenyl, 3-(2-methoxyethoxy)phenyl,
CA 02361285 2001-11-07
24
3,4-dipropoxyphenyl, 3-(difluoromethoxy)phenyl, 2-naphthyl, 3-
isopropoxyphenyl, 1-methyl-lH-indol-5-yl, 2,3-dihydro-l-benzofuran-5-yl, 1,3-
diethyl-2-oxo-2,3-dihydro-1 H-benzimidazol-5-yl, 3-(trifluoromethoxy)phenyl,
1-methyl-lH-indol-6-yl, 3-(cyclopropoxy)phenyl, 3-
(cyclopropylmethoxy)phenyl, 3-(difluoromethoxy)phenyl, 3-(1,1,2,2-
tetrafluoroethoxy)phenyl, 1-ethyl-IH-indol-5-yl, 3-(diethylamino)phenyl, 6-
methoxy-2-naphthyl, 3-[(methylsulfonyl)amino]phenyl, 3-
[methyl(methylsulfonyl)amino]phenyl, 3-[ethyl(methylsulfonyl)amino]phenyl,
1H-indol-5-yl, 3-fluoro-4-methoxyphenyl, and 3-(difluoromethyl)phenyl.
The R6 and R8 substituents for Formulae I-V above may be, but are not
limited to hydrogen, butyl, benzyl, benzyloxymethyl, ethyl, propyl,
~..,.
phenylsulfanylmethyl, benzylsulfanylmethyl, methylsulfanylethyl,
ethylsulfanylmethyl, methyl, or carboxyethyl.
R4 and R13 may be linked to form a ring such as 1-pyrrolidino, 1-
piperidino, 4-methyl-l-piperazino, 4-acetyl-l-piperazino and 4-morpholino
among others.
R6 and Rg may be linked to form a ring such as cyclopropyl, cyclobutyl,
cyclopentyl and cyclohexyl, among others.
R9 and R10 may be linked to form a ring such as cyclopropyl, cyclobutyl,
cyclopentyl and cyclohexyl, among others.
Abbreviations
Abbreviations which have been used in the schemes and the examples
which follow are: BOC for t-butyloxycarbonyl; EtOAc for ethyl acetate; DMF
for dimethylformamide; THF for tetrahydrofuran; Tos for p-toluenesulfonyl;
DCC for dicyclohexylcarbodiimide; HOBT for 1-hydroxybenzotriazole; TFAA
for trifluoroacetic anhydride; NMM for N-methyl morpholine; DIPEA for
diisopropylethylamine; DCM for methylene dichloride; LHMDS for lithium
hexamethyl disilazide; NaHMDS for sodium hexamethyl disilazide; CDI for
1,1'-carbonyldiimidazole HBTU for O-benzotriazol-l-yl-N,N,N', N'-
tetramethyluronium hexafluorophosphate, EDCI for 1-[3-(dimethylamino)
propyl]-3-ethylcarbodiimide hydrochloride and TBS for TRIS-buffered saline.
- - - --- - -----
CA 02361285 2001-11-07
Amino acids are abbreviated as follows: C for L-cysteine; D for L-aspartic
acid;
E for L-glutamic acid; G for glycine; H for L-histidine; I for L-isoleucine; L
for
L-leucine; N for L-asparagine; P for L-proline; Q for L-glutamine; S for L-
serine; T for L-threonine; V for L-valine, and W for L-tryptophan.
5 Examples of the procedures utilized to synthesize the compounds are
illustrated by the following schemes.
il
CIH3N COZMe 1) NaHCO3 Ne
2)CzH5CH0
\l~/~\ 2
Ph _^COC I
TEA, Et20
Ph N COZMe POCI3, DMF ph `
'"~ ^/N CO Me
0 75 C 0
\/\
4
3
NaOH 1CO zEt
HZN O>
O
\ 6 I
~-- CO2Et
Ph N CO2H I\ O
~ HBTU Ph N O
0 H
/
5 7
NaOH
Diastendmer 9 HPLC 0 COZH
a
Diastereomer 10 Ph N O
O H 0 >
8
10 Scheme 1
CA 02361285 2001-11-07
26
p COOEt p COOEt
f;30 BocHN~N p HZN~N p
H N O Boc-L-NIe-OH = H > HCl H z / HBTU, DIPEA 22 O Dioxane 23 O
6
H,N --_,OH
S CHO NaBH(OAc)3 S O NaBH4, MeOH S N
L HN~ ~~~OH
24 25
BOC a) DMSO, (COCI)2 Boc
B~ QCHO
Zp S ~~~O[{ b) 26
c) REt3 26 27
S NBoc p COOEt
N p HCI, dioxane
23 H
NaBH(OAc)3 0
28
S
\ NH O COOEt ^ C130 O
O CDI H ~
H
O> O 29 30
NaOH,THF Z
N ~ao /~ O H,O, MeOH ~`~ H IO\
C 20
Scheme 2
CA 02361285 2001-11-07
27
Scheme 3, shown below, illustrates the procedure described in Example 11.
O COOEt O COOEt
H2N"~- O ~ OEt H H >
Y
PhCH3, reflux
23 32
Me2N O COOEt
/ NaOH, THF
" /~CHO O HZO, MeOH
EtOH, AcOH 0 0 = H I, ~
Reflux
33
O COOH
Oyf ~
O O- H >
I~ O
C
31
Scheme 3
Scheme 4, shown below, illustrates Example 12.
oy~ 0 O 0 N~H NaBHy ~ N ~J H O
0 O p EtOH OH 0 \.-/\ 0 >
33 35
CO,Et
H,, Pd/C _ \ I N NaOH, THF
,,~-
EtOAc H
~ 0 H,O, MeOH
36
0 COOH
~
0 H ~ i ~
C
34
Scheme 4
CA 02361285 2001-11-07
28
Scheme 5, shown below, illustrates the procedure of Example 13.
Cl NOz
I ~ NO2
I
N O2N Cl-
I N+
Acetone, reflux
37
Q OH
HO NH K3Fe(CN)6, KOH,
Z CI- N~ water, toluene
n-Butanol I / ~ I
38
q --- OH COzH
RuC13, K2S208, N O/ KOH, water N O/
39 40
Scheme 5
Scheme 6, shown below, illustrates the procedure described in Example 14.
OH
~-, N O
OH
ci- I N 42
3Fe(CN)6, KOH,
K
~ o
/ water, toluene +
OH
41 O N
\ ~ \ I
43
Scheme 6
CA 02361285 2001-11-07
29
Scheme 7, shown below, illustrates the procedure described in Example 15.
O
~
~~ H
H, HZO
:z z OMe
HZN C1 O MeO
O 44
----~OMe S H
S ONa NH') H N OMe N N EDCI, HOBT C~ O
H NMM, DMF
46
s`/ 1
O
0
Br Br s NI'A Cs2CO3, THF
- ~ ~ N OMe
Et3N, CICHZCHZCI O~ 0 Br
47
s
O 0
O-J~ ~OMe
48
Scheme 7
CA 02361285 2001-11-07
Scheme 8, shown below, illustrates the procedure described in Example 16.
F NH2 NO2 H
NO ~ N Fe z
K2CO3, DMF 1~, AcOH/EtOH
49
NH2 H ~ I
N ~ CDI, CHZCIi
NNH
50 O
51
BrCH 2COOEt ~
C(--P O
'OEt
NaH, DMF u
I
~
O
52
Scheme 8
5 Scheme 9, shown below, illustrates the procedure described in Example 17.
O O
HN~'~ N
NH KZC03, DMF eNH
eO 0
53
Scheme 9
CA 02361285 2001-11-07
31
Scheme 10, shown below, illustrates the procedure described in Example 18.
Scheme 10
~ COOH
O
CIH H2N~ / NHZ cyAOMe
_ OMe EDCI, HOBt, NH2 0 NMM, DMF
54
\
I i YO
CDI 0
; C1CH2CHZCl, 85 C HNN,~,KOMe
0 55
Scheme 11, shown below, illustrates the procedure described in Example 19.
CHO
`''~ I~I N~
HN KOHJEtOH, 85 C
0 O
56
Scheme 11
Scheme 12, shown below, illustrates the procedure described in Example 20.
~LBr N
THF
NOZ NOz 57
Scheme 12
15
CA 02361285 2001-11-07
32
Scheme 13, shown below, illustrates the procedure described in Example
21.
0
crc: HSCHzCHZOH, HC1, EtOH, H20 N
58
Scheme 13
~.~_
CA 02361285 2001-11-07
33
Scheme 14, shown below, illustrates the procedure of Example 22.
O COOEt
a) iBuOCOC1, NMM, 0
Br OH THF, DMF, 0 C Br
O
b) q H ,
O
H2N O~
O 59
6
BF3=OEt2, Et3SiH,
CICH CH Cl,
I\ a) LDA, THF, - 78 C OY RT t Reflux
N b) PhCHO ' N
F OH F
a) NaHMDS
6 N HCl DME, -40 C
Reflux b) LiBr
N NH c) 59 in DMF,
F O -40 C to 45 C
61 62
COOEt
\ I I N~
o\
O
= H
~ O ,
63
5 Scheme 14
CA 02361285 2001-11-07
34
p O 0 0 a NaH THF, TM A
NaH,THF,BrBr RT
Oi - O
0 OC b. n-BuLi, -20 C
c. HCOzMe
64
O COCEt
H2N,J~ N O
O O O = H H I\ O COCEt
H O~ ~ O
\ MO~~ reflt>x 0 H O
+ /
65 r466
NaOH,THF
HzO, MeOH
H COQ-I H COQ-I
~ I I N O N O
H O + H O
O O O O
67 68
Scheme 15
~..:`
CA 02361285 2001-11-07
a) LDA, THF, -78 C HZ, Pd/C,
~ b) PhCHO, -78 C to RT MeOH, AcOH
~N OH N
F F
69
~ HCI, HZO
Dioxane, Reflux
N NH
F O
70 71
0--~ a) NaH, DMSO, RT b)
Fxhyl 2~ronmohexanoate qi
N COCEt
O
72
Scheme 16
~:-
CA 02361285 2001-11-07
36
Br EtOC)
q -~'COCEt I
~'2N~F
3
Pd(OAch, P(o-tAlYl)3 '~O NEt3, DMF,125 C 900C
73 74 CHO
EtOO COCEt
a) s-Buli, TF, -78 C
P N-H b) 170, TI-0;, -78 C PhN
Ph)
1~
75 CHF~ 76 F~
COCEt
H2, Pd/C, AcOH I I2N I~
EtOH, 35 C
77 CHFy
Scheme 17
~ _.
CA 02361285 2001-11-07
37
A detailed description of the preparation of representative compounds of
the present invention is set forth in the Examples.
The compounds of the present invention can be used in the form of
pharmaceutically acceptable salts derived from inorganic or organic acids. The
phrase "pharmaceutically acceptable salt" means those salts which are, within
the scope of sound medical judgement, suitable for use in contact with the
tissues of humans and lower animals without undue toxicity, irritation,
allergic
response and the like and are commensurate with a reasonable benefit/risk
ratio.
Pharmaceutically acceptable salts are well-known in the art. For example, S.
M.
Berge et al. describe phannaceutically acceptable salts in detail in J.
Pharmaceutical Sciences, 1977, 66: 1 et seq. The salts can be prepared in situ
~,-
during the final isolation and purification of the compounds of the invention
or
separately by reacting a free base function with a suitable organic acid.
Representative acid addition salts include, but are not limited to acetate,
adipate,.
alginate, citrate, aspartate, benzoate, benzene sulfonate, bisulfate,
butyrate,
camphorate, camphor sulfonate, digluconate, glycerophosphate, hemisulfate,
heptanoate, hexanoate, fumarate, hydrochloride, hydrobromide, hydroiodide, 2-
hydroxyethansulfonate (isothionate), lactate, maleate, methane sulfonate,
nicotinate, 2-naphthalene sulfonate, oxalate, palmitoate, pectinate,
persulfate, 3-
phenylpropionate, picrate, pivalate, propionate, succinate, tartrate,
thiocyanate,
phosphate, glutamate, bicarbonate, p-toluene sulfonate and undecanoate. Also,
the basic nitrogen-containing groups can be quaternized with such agents as
lower alkyl halides such as methyl, ethyl, propyl, and butyl chlorides,
bromides
and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl and diamyl
sulfates;
long chain halides such as decyl, lauryl, myristyl and stearyl chlorides,
bromides
and iodides; arylalkyl halides like benzyl and phenethyl bromides and others.
Water or oil-soluble or dispersible products are thereby obtained. Examples of
acids which can be employed to form pharmaceutically acceptable acid addition
salts include such inorganic acids as hydrochloric acid, hydrobromic acid,
sulphuric acid and phosphoric acid and such organic acids as oxalic acid,
maleic
acid, succinic acid and citric acid.
Basic addition salts can be prepared in situ during the final isolation and
purification of compounds of this invention by reacting a carboxylic acid-
CA 02361285 2001-11-07
38
containing moiety with a suitable base such as the hydroxide, carbonate or
bicarbonate of a pharmaceutically acceptable metal cation or with ammonia or
an organic primary, secondary or tertiary amine. Pharmaceutically acceptable
salts include, but are not limited to, cations based on alkali metals or
alkaline
earth metals such as lithium, sodium, potassium, calcium, magnesium and
aluminum salts and the like and nontoxic quaternary ammonia and amine
cations including ammonium, tetramethylammonium, tetraethylammonium,
methylammonium, dimethylammonium, trimethylammonium,
triethylammonium, diethylammonium, and ethylammonium among others.
Other representative organic amines useful for the formation of base addition
salts include ethylenediamine, ethanolamine, diethanolamine, piperidine,
piperazine and the like.
Dosage forms for topical administration of a compound of this invention
include powders, sprays, ointments and inhalants. The active compound is
mixed under sterile conditions with a pharmaceutically acceptable carrier and
any needed preservatives, buffers or propellants which can be required.
Opthalmic formulations, eye ointments, powders and solutions are also
contemplated as being within the scope of this invention.
Actual dosage levels of active ingredients in the pharmaceutical
compositions of this invention can be varied so as to obtain an amount of the
active compound(s) which is effective to achieve the desired therapeutic
response for a particular patient, compositions and mode of administration.
The
selected dosage level will depend upon the activity of the particular
compound,
the route of administration, the severity of the condition being treated and
the
condition and prior medical history of the patient being treated. However, it
is
within the skill of the art to start doses of the compound at levels lower
than
required to achieve the desired therapeutic effect and to gradually increase
the
dosage until the desired effect is achieved.
When used in the above or other treatments, a therapeutically effective
amount of one of the compounds of the present invention can be employed in
pure form or, where such forms exist, in pharmaceutically acceptable salt,
ester
or prodrug form. Altematively, the compound can be administered as a
pharmaceutical composition containing the compound of interest in combination
CA 02361285 2001-11-07
39
with one or more pharmaceutically acceptable excipients. The phrase
"therapeutically effective amount" of the compound of the invention means a
sufficient amount of the compound to treat disorders, at a reasonable
benefit/risk
ratio applicable to any medical treatment. It will be understood, however,
that
the total daily usage of the compounds and compositions of the present
invention will be decided by the attending physician within the scope of sound
medical judgement. The specific therapeutically effective dose level for any
particular patient will depend upon a variety of factors including the
disorder
being treated and the severity of the disorder; activity of the specific
compound
employed; the specific composition employed; the age, body weight, general
health, sex and diet of the patient; the time of administration, route of
administration, and rate of excretion of the specific compound employed; the
duration of the treatment; drugs used in combination or coincidental with the
specific compound employed; and like factors well known in the medical arts.
For example, it is well within the skill of the art to start doses of the
compound
at levels lower than required to achieve the desired therapeutic effect and to
gradually increase the dosage until the desired effect is achieved.
The total daily dose of the compounds of this invention administered to a
human or lower animal may range from about 0.0001 to about 1000 mg/kg/day.
For purposes of oral administration, more preferable doses can be in the range
of
~
from about 0.001 to about 5 mg/kg/day. If desired, the effective daily dose
can
be divided into multiple doses for purposes of administration; consequently,
single dose compositions may contain such amounts or submultiples thereof to
make up the daily dose.
The present invention also provides pharmaceutical compositions that
comprise compounds of the present invention formulated together with one or
more non-toxic pharmaceutically acceptable carriers. The pharmaceutical
compositions can be specially formulated for oral administration in solid or
liquid form, for parenteral injection or for rectal administration.
The pharmaceutical compositions of this invention can be administered
to humans and other mammals orally, rectally, parenterally, intracisternally,
intravaginally, intraperitoneally, topically (as by powders, ointments or
drops),
bucally or as an oral or nasal spray. The term "parenterally," as used herein,
CA 02361285 2001-11-07
refers to modes of administration which include intravenous, intramuscular,
intraperitoneal, intrasternal, subcutaneous and intraarticular injection and
infusion.
In another aspect, the present invention provides a pharmaceutical
5 composition comprising a component of the present invention and a
physiologically tolerable diluent. The present invention includes one or more
compounds as described above formulated into compositions together with one
or more non-toxic physiologically tolerable or acceptable diluents, carriers,
adjuvants or vehicles that are collectively referred to herein as diluents,
for
10 parenteral injection, for intranasal delivery, for oral administration in
solid or
liquid form, for rectal or topical administration, or the like.
The compositions can also be delivered through a catheter for local
delivery at a target site, via an intracoronary stent (a tubular device
composed of
a fine wire mesh), or via a biodegradable polymer. The compounds may also be
15 complexed to ligands, such as antibodies, for targeted delivery.
Compositions suitable for parenteral injection may comprise
physiologically acceptable, sterile aqueous or nonaqueous solutions,
dispersions,
suspensions or emulsions and sterile powders for reconstitution into sterile
injectable solutions or dispersions. Examples of suitable aqueous and
20 nonaqueous carriers, diluents, solvents or vehicles include water, ethanol,
polyols (propyleneglycol, polyethyleneglycol, glycerol, and the like),
vegetable
oils (such as olive oil), injectable organic esters such as ethyl oleate, and
suitable mixtures thereof.
These compositions can also contain adjuvants such as preserving,
25 wetting, emulsifying, and dispensing agents. Prevention of the action of
microorganisms can be ensured by various antibacterial and antifungal agents,
for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It
may
also be desirable to include isotonic agents, for example sugars, sodium
chloride
and the like. Prolonged absorption of the injectable pharmaceutical form can
be
30 brought about by the use of agents delaying absorption, for example,
aluminum
monostearate and gelatin.
Suspensions, in addition to the active compounds, may contain
suspending agents, as for example, ethoxylated isostearyl alcohols,
CA 02361285 2001-11-07
41
polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose,
aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of
these substances, and the like.
Proper fluidity can be maintained, for example, by the use of coating
materials such as lecithin, by the maintenance of the required particle size
in the
case of dispersions and by the use of surfactants.
In some cases, in order to prolong the effect of the drug, it is desirable to
slow
the absorption of the drug from subcutaneous or intramuscular injection. This
can be accomplished by the use of a liquid suspension of crystalline or
amorphous material with poor water solubility. ThL rate of absorption of the
drug then depends upon its rate of dissolution which, in tum, may depend upon
crystal size and crystalline form. Alternatively, delayed absorption of a
parenterally administered drug form is accomplished by dissolving or
suspending the drug in an oil vehicle.
Injectable depot forms are made by forming microencapsule matrices of
the drug in biodegradable polymers such as polylactide-polyglycolide.
Depending upon the ratio of drug to polymer and the nature of the particular
polymer employed, the rate of drug release can be controlled. Examples of
other biodegradable polymers include poly(orthoesters) and poly(anhydrides).
Depot injectable formulations are also prepared by entrapping the drug in
liposomes or microemulsions which are compatible with body tissues.
The injectable formulations can be sterilized, for example, by filtration
through
a bacterial-retaining filter or by incorporating sterilizing agents in the
form of
sterile solid compositions which can be dissolved or dispersed in sterile
water or
other sterile injectable medium just prior to use.
Solid dosage forms for oral administration include capsules, tablets,
pills, powders and granules. In such solid dosage forms, the active compound
may be mixed with at least one inert, pharmaceutically acceptable excipient or
carrier, such as sodium citrate or dicalcium phosphate and/or a) fillers or
extenders such as starches, lactose, sucrose, glucose, mannitol and silicic
acid;
b) binders such as carboxymethylcellulose, alginates, gelatin,
polyvinylpyrrolidone, sucrose and acacia; c) humectants such as glycerol; d)
disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca
CA 02361285 2001-11-07
42
starch, alginic acid, certain silicates and sodium carbonate; e) solution
retarding
agents such as paraffin; f) absorption accelerators such as quaternary
anunonium
compounds; g) wetting agents such as cetyl alcohol and glycerol monostearate;
h) absorbents such as kaolin and bentonite clay and i) lubricants such as
talc,
calcium stearate, magnesium stearate, solid polyethylene glycols, sodium
lauryl
sulfate and mixtures thereof. In the case of capsules, tablets and pills, the
dosage form may also comprise buffering agents.
Solid compositions of a similar type may also be employed as fillers in
soft and hard-filled gelatin capsules using such excipients as lactose or milk
sugar as well as high molecular weight polyethylene glycols and the like.
The solid dosage forms of tablets, dragees, capsules, pills and granules can
be
prepared with coatings and shells such as enteric coatings and other coatings
well-known in the pharmaceutical formulating art. They may optionally contain
opacifying agents and may also be of a composition such that they release the
active ingredient(s) only, or preferentially, in a certain part of the
intestinal tract,
optionally, in a delayed manner. Examples of embedding compositions which
can oe used include polymeric substances and waxes.
The active compounds can also be in micro-encapsulated form, if
appropriate, with one or more of the above-mentioned excipients.
Liquid dosage forms for oral administration include pharmaceutically
acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition
to
the active compounds, the liquid dosage forms may contain inert diluents
commonly used in the art such as, for example, water or other solvents,
solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol,
ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene
glycol, 1,3-butylene glycol, dimethyl formamide, oils (in particular,
cottonseed,
groundnut, corn, germ, olive, castor and sesame oils), glycerol,
tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of
sorbitan
and mixtures thereof.
Besides inert diluents, the oral compositions may also include adjuvants
such as wetting agents, emulsifying and st:-pending agents, sweetening,
flavoring and perfuming agents.
CA 02361285 2008-10-23
43
Compositions for rectal or vaginal administration are preferably
suppositories which can be prepared by mixing the compounds of this invention
with suitable non-irritating excipients or carriers such as cocoa butter,
polyethylene glycol or a suppository wax which are solid at room temperature
but liquid at body temperature and therefore melt in the rectum or vaginal
cavity
and release the active compound.
Compounds of the present invention can also be administered in the
form of liposomes. As is known in the art, liposomes are generally derived
from phospholipids or other lipid substances. Liposomes are formed by mono-
or multi-lamellar hydrated liquid crystals which are dispersed in an aqueous
medium. Any non-toxic, physiologically acceptable and metabolizable lipid
capable of forming liposomes can be used. The present compositions in
liposome form can contain, in addition to a compound of the present invention,
stabilizers, preservatives, excipients and the like. The preferred lipids are
natural and synthetic phospholipids and phosphatidyl cholines (lecithins) used
separately or together.
Methods to form liposomes are known in the art. See, for example,
Prescott, Ed., Methods in Cell Biolo~y, Volume XIV, Academic Press, New
York, N.Y. (1976), p. 33 et seq.
The term "pharmaceutically acceptable prodrugs" as used herein
represents those prodrugs of the compounds of the present invention which are,
within the scope of sound medical judgement, suitable for use in contact with
the tissues of humans and lower animals without undue toxicity, irritation,
allergic response, and the like, commensurate with a reasonable benefit/risk
ratio, and effective for their intended use, as well as the zwitterionic
forms,
where possible, of the compounds of the invention. Prodrugs of the present
invention may be rapidly transformed in vivo to the parent compound of the
above formula, for exampie, by hydrolysis in blood. A thorough discussion is
provided in T. Higuchi and V. Stella, Pro-dru s as Novel Delive Systems, V.
14 of the A.C.S. Symposium Series, and in Edward B. Roche, ed., Bioreversible
Carriers in Drug Design, American Pharmaceutical Association and Pergamon
Press (1987).
CA 02361285 2001-11-07
44
Compounds of the present invention that are formed by in vivo
conversion of a different compound that was administered to a manunal are
intended to be included within the scope of the present invention.
Compounds of the present invention may exist as stereoisomers wherein
asymmetric or chiral centers are present. These stereoisomers are "R" or "S"
depending on the configuration of substituents around the chiral carbon atom.
The present invention contemplates various stereoisomers and mixtures thereof.
Stereoisomers include enantiomers and diastereomers, and mixtures of
enantiomers or diastereomers. Individual stereoisomers of compounds of the
present invention may be prepared synthetically from commercially available
starting materials which contain asymmetric or chiral centers or by
preparation
of racemic mixtures followed by resolution well-known to those of ordinary
skill in the art. These methods of resolution are exemplified by (1)
attachment
of a mixture of enantiomers to a chiral auxiliary, separation of the resulting
mixture of diastereomers by recrystallization or chromatography and liberation
of the optically pure product from the auxiliary or (2) direct separation of
the
mixture of optical enantiomers on chiral chromatographic columns.
The compounds of the invention can exist in unsolvated as well as
solvated forms, including hydrated forms, such as hemi-hydrates. In general,
the solvated forms, with pharmaceutically acceptable solvents such as water
and
ethanol among others are equivalent to the unsolvated forms for the purposes
of
the invention. In another aspect, the present invention contemplates a process
of
inhibiting the binding of a4P , integrin to VCAM-l. A process of the present
invention can be used either in vitro or in vivo. In accordance with a process
of
the present invention, a cell expressing a,,P i integrin is exposed to a cell
expressing VCAM-1 in the presence of an effective inhibiting amount of a
compound of the present invention.
A cell expressing aj , integrin can be a naturally occurring white blood
cell, mast cell or other cell type that naturally expresses a4(3, on the cell
surface,
or a cell transfected with an expression vector that contains a poly-
nucleotide
(e.g., genomic DNA or cDNA) that encodes a4P , integrin. In an especially
preferred embodiment, a.,Pi integrin is present on the surface of a white
blood
CA 02361285 2001-11-07
cell such as a monocyte, a lymphocyte or a granulocyte (e.g., an eosinophil or
a
basophil).
A cell that expresses VCAM-1 can be a naturally occurring cell (e.g. an
endothelial cell) or a cell transfected with an expression vector containing a
5 polynucleotide that encodes VCAM-l. Methods for producing transfected cells
that express VCAM-1 are well known in the art.
Where VCAM-1 exists on the surface of cell, the expression of that
VCAM-1 is preferably induced by inflammatory cytokines such as tumor
necrosis factor-a, interleukin-4 and interleukin-1(3.
10 Where the cells expressing a4p, integrin and VCAM-l are in a living
organism, a compound of the present invention is administered in an effective
amount to the living organism. Preferably, the compound is in a pharmaceutical
composition of this invention. A process of the present invention is
especially
useful in treating diseases associated with uncontrolled migration of white
blood
15 cells to damaged tissue. Such diseases include, but are not limited to,
asthma,
atherosclerosis, rheumatoid arthritis, allergy, multiple sclerosis, lupus,
inflammatory bowel disease, graft rejection, contact hypersensitivity, type I
diabetes, leukemia, and brain cancer. Administration is preferably
accomplished via intravascular, subcutaneous, intranasal, transdermal or oral
20 delivery.
The present invention also provides a process of selectively inhibiting
the binding of a4(3, integrin to a protein comprising exposing the integrin to
the
protein in the presence of an effective inhibiting amount of a compound of the
present invention. In a preferred embodiment, the a4(3, integrin is expressed
on
25 the surface of a cell, either naturally occurring or a cell transformed to
express
a4(3, integrin.
The protein to which the a,p , integrin binds can be expressed'either on a
cell surface or be part of the extracellular matrix. Especially preferred
proteins
are fibronectin or invasin.
30 The ability of compounds of the present invention to inhibit binding is
described in detail hereinafter in the Examples. These Examples are presented
to describe preferred embodiments and utilities of the invention and are not
CA 02361285 2001-11-07
46
meant to limit the invention unless otherwise stated in the claims appended
hereto.
Example 1
Compound 8, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-benzyl-5-
methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid, of the structure
shown below, was synthesized as follows.
~ O C02H
Ph N O
O H
O >
8
The structures of the compounds identified by number in this Example are
found in Scheme I above.
Step 1: A solution of 540 mg of 2-aminohexanoic acid methyl ester
hydrochloride salt 1 in 20 ml of methylene chloride was washed with excess
saturated sodium bicarbonate. The organic layer was separated, dried over
magnesium sulfate, and concentrated in vacuo to give 365 mg of 2-
aminohexanoic acid methyl ester as a colorless oil. This material was combined
with 5 ml of benzene, 0.28 ml of propionaldehyde, and excess magnesium
sulfate. After stirring for 15 minutes, the reaction mixture was filtered and
concentrated in vacuo to yield 420 mg of compound 2 as a colorless oil.
Compound 2 was used directly without further purification.
Step 2: To an ice-bath cooled solution of 1050 mg of compound 2 in 10
ml of diethyl ether, under a positive nitrogen atmosphere, was added 0.80 ml
of
triethylamine, and a solution of 964 mg of 3-phenylpropanoyl chloride in 2 ml
of diethyl ether. The ice bath was removed and the reaction mixture stirred
for
30 minutes. The reaction mixture was then concentrated in vacuo and the
residual materials further separated by silica gel chromatography using 15%
ethyl acetate / hexane as the eluant to yield 468 mg of compound 3 as a
colorless
CA 02361285 2001-11-07
47
oil. Compound 3: 'H NMR (CDC 13): S 0.87 (t, J = 7.0 Hz, 3H), 1.26 (m, 4H),
1.68 (dd, J = 7.0, 1.1 Hz, 3H), 1.74 (m, 1H), 1.97 (m, 1H), 2.70 (t, J = 7.9
Hz,
2H), 2.96 (t, J = 7.9 Hz, 2H), 3.68 (s, 3H), 4.96 (dd, J. = 10.1, 5.3 Hz, 1H),
5.32
(dq, J = 13.9, 7.0 Hz, 1H), 6.13 (dd, J = 13.9, 1.1 Hz, 1H), 7.20 (m, 2H),
7.25
(m, 3H).
Step 3: N,N-Dimethylformamide (1.63 ml) was added dropwise to an
ice-cooled flask containing 4.57 ml of phosphorus oxychioride sealed under a
positive nitrogen atmosphere. After 5 minutes, the reaction solution was
cannulated into a flask containing 2.22 gm of compound 3. This mixture was
stirred at room temperature under a positive nitrogen atmosphere for 2 hours
and then heated at 75 C for 46 hours. The dark-colored reaction mixture was
poured over ice and mixed with an excess of sodium bicarbonate and ethyl
acetate. The mixture was saturated with sodium chloride and the organic layer
separated. The aqueous layer was extracted (3 X 100 ml) with ethyl acetate.
The combined organic materials were dried over magnesium sulfate and
concentrated in vacuo to yield 1.70 gm of a dark-colored oil. Methylene
chloride extraction (3 X ) of the aqueous layer yielded an additional 200 mg
of
material after drying (MgSOa) and condensation in vacuo. The combined
residual oils were further purified by silica gel chromatography using 20% -
25%
ethyl acetate / hexane as the eluant to yield 815 mg of compound 4 as a yellow
oil. Compound 4: 'H NMR (CDCl3): S 0.87 (t, J = 7.2 Hz, 3H), 1.18 (m, 1H),
1.31 (m, 3H), 1.87 (m, 1H), 2.00 (d, J = 0.7 Hz, 3H), 2.16 (m, IH), 3.72 (s,
3H),
3.85 (br. s, 2H), 5.57 (dd, J = 10.1, 5.7 Hz, 1H), 6.82 (br. s, 1H), 6.94 (br.
s,
IH), 7.23 (m, 3H), 7.30 (m, 2H).
Step 4: To a solution of 86 mg of compound 4 in 3 ml of
tetrahydrofuran was added I ml of 2N sodium hydroxide and 2 ml of methanol.
After complete hydrolysis, the reaction mixture was acidified with 2N
hydrochloric acid and saturated with sodium chloride. The mixture was
extracted (3X) with ethyl acetate and the combined extracts were dried with
magnesium sulfate and concentrated in vacuo to yield 80 mg of compound 5 as
a light yellow oil. Compound 5: 'H NMR (CDC13): S 0.88 (t, J = 7.1 Hz, 3H),
1.18 (m, 1H), 1.33 (m, 3H), 2.04 (d, J= 0.7 Hz, 3H), 2.07 (m, 1H), 2.27 (m,
CA 02361285 2001-11-07
48
1H), 3.86 (d, J = 16.1 Hz, 1H), 3.90 (d, J = 16.1 Hz, 1H), 5.04 (dd, J = 9.0,
6.8
Hz, 1H), 6.96 (br. s, 1H), 6.98 (br. s, IH), 7.23 (m, 3H), 7.31 (m, 2H).
Step 5: To a solution of 80 mg of compound 5 in I ml of N,N-
dimethylformamide at room temperature and under a positive nitrogen
atmosphere, was added 78 mg of (S)-compound 6, 0.057 ml of
diisopropylethylamine, and 137 mg of HBTU. The mixture was stirred for 16
hours and then mixed with 1:1 ethyl acetate / hexane. This mixture was washed
with 2N hydrochloric acid, saturated sodium bicarbonate, water (2X), and
finally brine. The resulting solution was dried over magnesium sulfate and
concentrated in vacuo to yield 156 mg of a yellow o:l. This material was
further
purified by silica gel chromatography using 25% ethyl acetate as the eluant to
give 109 mg of compound 7 as a colorless oil. Compound 7: (least polar
diastereomer): 'H NMR (CDC13): 6 0.85 (t, J = 7.1 Hz, 3H), 1.11 (t, J =7.1 Hz,
3H), 1.18 (m, 1H), 1.30 (m, 3H), 1.78 (m, 1H), 2.02 (d, J = 0.8 Hz, 3H), 2.14
(m, 1H), 2.57 (dd, J = 15.4, 7.1 Hz, 1H), 2.66 (dd, J = 15.4, 6.6 Hz, 1H),
3.86
(br. s, 2H), 3.95 (q, J = 7.1 Hz, 2H), 5.17 (m, 111), 5.42 (t, J= 7.7 Hz, 1H),
5.93
(s, 2H), 6.72 (m, 2H), 6.74 (m, 1 H), 6.90 (m, 1 H), 7.11 (br. s, l H), 7.23
(m,
3H), 7.30 (m, 2H), 7.37 (d, J = 7.7 Hz, IH).
Step 6: A solution composed of 109 mg of compound 7, 3 ml of
tetrahydrofuran, 1 ml of 2N sodium hydroxide, and 2 ml of methanol was stirred
at room temperature until hydrolysis was complete. The mixture was then
diluted with water and extracted with diethyl ether. The aqueous layer was
acidified with 2N hydrochloric acid and extracted (3X) with ethyl acetate. The
combined extracts were dried with magnesium sulfate and concentrated in
vacuo to yield 103 mg of compound 8, a 1:1 diasteroisomeric mixture, as an off-
white foam.
The diastereomeric mixture was separated by reverse-phase HPLC using
a 30 -55% acetonitrile / water gradient to yield Compound 9 (R,S) and
Compound 10 (S,S).
Compound 9 (most polar diastereomer): 'H NMR (CD3SOCD3): 8 0.83
(t, J = 7.1 Hz, 3H), 1.13 (m, 2H), 1.26 (m, 2H), 1.76 (m, 1H), 1.96 (s over-
lapping m, 4H), 2.62 (dd, J = 15.8, 6.6 Hz, 1 H), 2.70 (dd, J = 15.8, 8.4 Hz,
1 H),
CA 02361285 2001-11-07
49
3.69 (d, J = 14.8 Hz, 1 H), 3.73 (d. J= 14.8 Hz, 1 H), 5.09 (m, IH), 5.47 (dd,
J
9.2, 6.6 Hz, 1H), 6.71 (dd, J = 8.0, 1.5 Hz, 1H), 6.77 (d, J= 8.0 Hz, 1H),
6.84
(d, J= 1.5 Hz, 1H), 7.00 (d, J = 1.8 Hz, IH), 7.14-7.30 (m, 6H), 8.70 (d, J =
8.1
Hz, 1H).
Compound 10: (least polar diastereomer) NMR (CD3SOCD3): 6 0.76 (t,
J = 7.3 Hz, 3H), 1.01 (m, 2H), 1.20 (m, 2H), 1.98 (br. s, 3H), 2.60 (dd, J =
15.8,
7.0 Hz, 1H), 2.68 (dd, J=15.8, 7.7 Hz, 1H), 3.71 (d, J= 15.0 Hz, 1H), 3.76 (d,
J
= 15.0 Hz, 1 H), 5.05 (ddd, J = 8.0, 7.7, 7.0 Hz, IH), 5.51 (dd, J = 9.2, 6.6
Hz,
1 H), 5.98 (s, 2H), 6.77 (dd, J 8.0, 1.4 Hz, 1 H), 6.83, (d, J= 8.0 Hz, IH),
6.89
(d, J = 1.4 Hz, IH), 7.02 (d, J 2.2 Hz, 1 H), 7.18 (m, 1 H), 7.25 (m, 4H),
7.38
(m, 1 H), 8.79 (d, J = 8.0 Hz, IH), 12.08 (br. s, IH).
F._
Example 2
Compound 12, (3S)-3-((2R, S)-2-(3-benzyl-5-methyl-2-oxo-1(2H)-
pyridinyl)hexanoylamino)-3-(2,3-dihydro-l-benzofuran-5-yl)propanoic acid,
shown below, was synthesized according to the procedure of Example 1,
except that compound A, shown below, was substituted for compound 6 in step
5.
/ ~ O CO zH
\ I I N
N
O H O
!T
C02Et
H2N 1
O
A
Example 3
Compound 13, (3S)-3-((2R,S)-2-(3-benzyl-5-methyl-2-oxo-1(2H)-
pyridinyl)hexanoylamino)-3-(4-methylphenyl)propanoic acid, shown below,
was obtained by the procedure of Example 1,
CA 02361285 2001-11-07
N~ O CO2H
~ I I N \
O H
CH3
13
except that compound B, shown below, was substituted for compound 6 in step
5.
C02Et
H2N
CH3
B
Example 4
5 Compound 14, (3S)-3-((2R,S)-2-(3-benzyl-5-methyl-2-oxo-1(2H)-
pyridinyl)hexanoylamino)-3-(4-fluorophenyl)propanoic acid, shown below was
obtained by the procedure of Example 1,
\ O C02H
N
~ I I \
o H
F
14
except that compound 11, shown below, was substituted for compound 6 in step
10 5.
C02Et
H2N
F
Jl
CA 02361285 2001-11-07
51
Example 5
Compound 15, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-(4-
methoxybenzyl)-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic
acid, shown below, can be obtained by the procedure of Example 1,
H3CO / ~ O CO2H
\ I I N N O
H >
O
O
5
except that 3-(4-methoxyphenyl)-propanoyl chloride should be substituted for 3-
phenylpropanoyl chloride in step 2.
Example 6
Compound 16, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-(4-
10 methylbenzyl)-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid,
shown below can be obtained by the procedure of Example 1,
H3C O CO2H
\ I I
N N O
O H
O >
16
except that 3-(4-methylphenyl)-propanoyl chloride should be substituted for 3-
phenylpropanoyl chloride in step 2.
15 Example 7
Compound 17, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-(4-
fluorobenzyl)-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid,
shown below, was obtained by the procedure of Example 1,
F /
O CO2H
\ ( I N N O
O H
O ~
17
CA 02361285 2001-11-07
52
except that 3-(4-fluorophenyl)-propanoyl chloride was substituted for 3-
phenylpropanoyl chloride in step 2.
Example 8
Compound 18, (3S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-(4-
chlorobenzyl)-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid,
shown below was obtained by the procedure of Example 1,
CI O C02H
\ N \ O
O H
O >
18
except that 3-(4-chlorophenyl)-propanoyl chloride was substituted for 3-
phenylpropanoyl chloride in step 2.
Example 9
Compound 19, ()S)-3-(1,3-benzodioxol-5-yl)-3-((2R,S)-2-(3-(3-
chlorobenzyl)-5-methyl-2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid,
shown below was obtained by the procedure of Example 1,
CI
O C02H
I I ~
N N ~
O H
O
19
except that 3-(3-chlorophenyl)-propanoyl chloride was substituted for 3-
phenylpropanoyl chloride in step 2.
Example 10
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[2-oxo-3-(2-
thienylmethyl)-1-imidazolidinyl]hexanoyl}amino)propanoic acid (20).
CA 02361285 2001-11-07
53
Step One: To a solution of 6 (680 mg, 2.87 mmol) and N-tert-
butoxycarbonyl-L-norleucine (696 mg, 3.01 mmol) in DMF (14.4 ml) at room
temperature under a dry nitrogen atmosphere, N, N-diisopropylethylamine (0.52
ml, 3.0 mmol) and HBTU (1.25 g, 3.3 mmol) were added sequentially. The
resulting mixture was stirred at room temperature overnight then was diluted
with a 1:1 mixture of hexanes:ethyl acetate and washed 2N HC1, H20, saturated
NaHCO3, HZO (3X) and brine. The organic phase was dried over MgSO4 and
filtered and the filtrate was concentrated under reduced pressure to give 22
(1.27g, 98%) as a light yellow solid.
Sten Two: To a flask containing 22 (1.27g, 2.82 mmol) sealed with a
rubber septum at room temperature under a dry nitrogen atmosphere, HCl (7.2
ml, 4.OM in dioxane, 28.8 mmol) was added by syringe. The nitrogen needle
was removed and the mixture in the sealed flask was stirred for 1 hour. The
mixture was diluted with CH,C12 and washed with saturated NaHCO3. The
organic phase was dried over MgSO4 and filtered and the fi(trate was
concentrated under reduced pressure to give 23 (892 mg, 90%) as a light yellow
oil.
Step Three: To a solution of ethanolamine (1.70 g, 27.8 mmol) and 2-
thiophenecarboxaldehyde (0.52 ml, 5.6 mmol) in 1,2-dichloroethane (22 ml) at
room temperature under a dry nitrogen atmosphere, sodium
triacetoxyborohydride (1.66 g, 7.8 mmol) was added. The resulting mixture was
stirred at room temperature overnight then was diluted with CHZCIz and washed
with a 1:1 mixture of saturated NaHCO3 and brine. The aqueous phase was
extracted with CHZCIZ and the organic phases were combined, dried over
MgSO4 and filtered. The filtrate was concentrated under reduced pressure to
give 24 (840 mg, 97%) as a pale yellow oil.
Step Four: A solution of aminal 24 (840 mg, 5.41 mmol) in methanol
(10 ml) was stirred at room temperature for 30 minutes, cooled to 0 C and then
sodium borohydride (106 mg, 2.8 mmol) was added. The mixture was allowed
to warm to room temperature and was then stirred for 1 hour. The mixture was
quenched by dropwise addition of water then diluted with CH2Clz and a 1:1
mixture of saturated NaHCO3 and brine. The aqueous phase was extracted with
CA 02361285 2001-11-07
54
CH2C12 (2X) and the organic phases were combined, dried over MgSO, and
filtered. The filtrate was concentrated under reduced pressure to give 25 (420
mg, 49%) as a pale yellow viscous oil.
Step Five: A solution of 25 (420 mg, 2.67 mmol) and di-tert-butyl
dicarbonate (650 mg, 2.98 mmol) in CH2C12 (10 ml) was stirred at room
temperature under a dry nitrogen atmosphere for 20 minutes and then
concentrated. The residue was filtered though silica gel, eluting with 7:3
hexanes:ethyl acetate increasing to 3:2 hexanes:ethyl acetate to yield 26 (610
mg, 88%) as a colorless, viscous oil.
Step Six: To a solution of methylsulfoxide (0.49 ml, 6.9 mmol) in
CHzCI, (11 ml) cooled to -78 C under a dry nitrogen atmosphere, oxalyl
chloride (1.7 ml, 2.0 M in CH2C12, 3.4 nunol) was added by syringe. The
resulting mixture was stirred at -78 C for 15 minutes, then a solution of 26
(590 mg, 2.3 mmol) in CH,CIz (10 ml) was added by cannula along with a
CH,C12 (5 ml) rinse. The mixture was stirred at -78 C for 30 minutes,
triethylamine (0.96 ml, 6.9 mmol) was added and the mixture was allowed to
warm to room temperature. The mixture was diluted with CH2CI, and washed
with saturated NaHCO. The organic phase was dried over MgSO4and filtered
and the filtrate was concentrated under reduced pressure to give 27 (630 mg)
as
a light yellow oil. This material was used without purification.
Step Seven: To a solution of 27 (102 mg, 0.40 mmol) and 23 (140 mg,
0.40 mmol) in 1,2-dichloroethane (4 ml) at room temperature under a dry
nitrogen atmosphere, sodium triacetoxyborohydride (119 mg, 0.56 mmol) was
added. The resulting mixture was stirred for 2 hours, then was diluted with
ethyl acetate and washed with saturated NaHCO3 and brine. The organic phase
was dried over MgSO, and filtered and the filtrate was concentrated under
reduced pressure to give 28 (232 mg) as a light yellow oil. This material was
used without purification.
Step Eight: To a flask containing 28 (232 mg crude material, 0.40 mmol
theoretical from previous step) sealed with a rubber septum at room
temperature
under a dry nitrogen atmosp:iere, HCl (1.95 ml, 4.OM in dioxane, 7.8 nunol)
was added by syringe. The nitrogen needle was removed and the mixture in the
CA 02361285 2001-11-07
sealed flask was stirred for 15 minutes. The mixture was diluted with CHZCIZ
and washed with a 1:1 mixture of saturated NaHCO3:brine. The organic phase
was dried over MgSO4 and filtered and the filtrate was concentrated under
reduced pressure to give 29 (180 mg) as a light yellow oil. This material was
5 used without purification.
SteQNine: To a solution of 29 (180 mg crude material, 0.40 mmol
theoretical from previous step) in 1,2-dichloroethane (3.7 ml) at room
temperature under a dry nitrogen atmosphere, carbonyldiimidazole (66 mg, 0.41
mmol) was added. The mixture was heated to 50 C (oil bath temperature) for 1
10 hour, the was concentrated. The residue was taken up in ethyl acetate and
washed with 2N HCI, H20, saturated NaHCO3 and brine. The organic phase
was dried over MgSO4 and filtered and the filtrate was concentrated under
reduced pressure. The residue was filtered through silica gel, eluting with
3:2
hexanes:ethyl acetate increasing to 1:1 hexanes:ethyl acetate to yield 30 (114
15 mg, 55% for 3 steps) as a colorless oil.
Step Ten: To a solution of 30 (114 mg, 0.22 mmol) in THF (3 ml) at
room temperature, NaOH (1 ml, 2N in H,O, 2 mmol) and methanol (enough to
give a clear solution, approximately 2 ml) were added. The resulting mixture
was stirred for 15 minutes, then was diluted with water and extracted with
ether.
20 The aqueous phase was acidified with HCI (2N) and extracted with ethyl
~_. acetate. The ethyl acetate layer was washed with brine, dried over MgSO4
and
filtered and the filtrate was concentrated under reduced pressure to give 20
(111
mg, 100%) as a white foam. `H NMR (400 MHz, CD3SOCD3) S 0.82 (t, J = 7.3
Hz, 3H), 1.09 (m, 2H), 1.25 (m, 2H), 1.48 (m, 1H), 1.64 m, 1H), 2.61 (dd, J =
25 15.8, 7.0 Hz, IH), 2.70 (dd, J = 15.8, 8.0 Hz, IH), 3.20 (m, 3H), 3.50 (m,
1H),
4.27 (dd, J= 9.5, 5.9 Hz, IH), 4.38 (d, J = 15.6 Hz, IH), 4.54 (d, J = 15.6
Hz,
IH), 5.08 (ddd, J = 8.1, 8.0, 7.0 Hz, 1 H), 5.91 (s, 2H), 6.76 (dd, J = 8.0,
1.5 Hz,
1 H), 6.83 (d, J = 8.0 Hz, 1 H), 6.88 (d, J = 1.5 Hz, 1 H), 6.99 (nl, 2H),
7.43 (dd, J
= 4.4, 1.8 Hz, 1H), 8.43 (d, J = 8.1 Hz, 1H).
30 Synthetic procedures similar to those described above may be utilized to
obtain the following compounds: (3S)-(1,3-benzodioxol-5-yl)-3-(((2S)-2-(2-
oxo-3-(2-thienylmethyl)tetrahydro-1(2H)-pyrimidinyl)hexanoyl)amino)
CA 02361285 2001-11-07
56
propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-(((2S)-2-(2-oxo-4-(2-thienyl)-
3-(2-thienylmethyl)tetrahydro-1((2H)-pyrimidinyl)hexanoyl)amino)propanoic
acid and (3S)-3-(1,3-benzodioxol-5-yl)-3-(((2S)-2-(2-oxo-3-(2-thienylmethyl)-
1,3-diazepan-1-yl)hexanoyl)amino)propanoic acid.
Example 11
Synthesis of (3 S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[2-oxo-3-
(phenylcarbonyl)-1(2H)-pyridinyl]hexanoyl}amino)propanoic acid (31).
O
O q N O
p p >
O
H3C
31
Step One: A solution of 23 (541 mg, 1.54 mmol) and ethyl
benzoylacetate (0.53 mL, 3.09 mmol) in toluene (15 mL) was heated to reflux
for 2 hours. The resulting mixture was cooled to room temperature and
concentrated under reduced pressure. The residue was recrystallized from
hexanes/CHzCIZ to give compound 32 (310 mg, 40%) as a pale yellow solid.
Step Two: To a suspension of 32 (851 mg, 1.71 ranol) in ethanol
(absolute, 6.8 mL) and acetic acid (glacial, 0.34 mL) at room temperature
under
nitrogen, 3-(dimethylamino)acrolein (1.02 mL, 10.2 mmol) was added by
syringe. The resulting mixture was heated to reflux overnight, cooled to room
temperature and diluted with ethyl acetate. This mixture was washed with HC1
(2N, twice) and brine. The organic phase was dried over MgSO4 and filtered
and the filtrate was concentrated under reduced pressure. The residue was
purified by silica gel chromatography, eluting with 3:2 hexanes:ethyl acetate
to
give 33 (476 mg, 52%) as a light yellow oil.
Step Three: To a solution of 33 (115 mg, 0.22 mmol) in THF (6 mL) at
room temperature, aqueous NaOH (2N, 2 mL) and methanol (4 mL) were added.
The resulting solution was stirred for 15 minutes, diluted with water and
CA 02361285 2001-11-07
57
extracted with Et20. The aqueotis phase was acidified with HCl (2N) and was
extracted with ethyl acetate. The ethyl acetate layer was washed with water
and
brine, dried over MgSO4 and filtered. The filtrate was concentrated under
reduced pressure to give (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[2-oxo-3-
(phenylcarbonyl)-1(2H)-pyridinyl]hexanoyl}amino)propanoic acid (31, 100 mg,
92%) as a pale yellow foam. 'H NMR (400 MHz, CD3SO2CD3): 6 0.81 (t, J =
7.3 Hz, 3H), 1.08 (m, 2H), 1.25 (m, 2H), 1.80 (m, 1H), 1.93 (m, IH), 2.61 (dd,
J
= 15.8, 6.8 Hz, 1 H), 2.68 (dd, J = 15.8, 7.9 Hz, IH), 5.09 (m, 1 H), 5.49
(dd, J =
9.5, 6.2 Hz, 1H), 5.98 (s, 2H), 6.24 (t, J = 7.0 Hz, IH), 6.78 (dd, J = 8.1,
1.4 Hz,
IH), 6.84 (d, J = 8.1 Hz, IH), 6.89 (d, J = 1.4 Hz, 1 H), 7.49 (t, J = 7.7 Hz,
2H),
7.62 (m, 1 H), 7.70 (m, 3H), 7.97 (dd, J = 7.0, 2.2 Hz, 1 H), 8.87 (d, J = 8.1
Hz,
IH), 12.11 (br. s, 1 H).
Example 12
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[2-oxo-3-
(phenylmethyl)-1(2H)-pyridinyl]hexanoyl}amino)propanoic acid (34).
O
O OH
-~'AN \ O
0 >
H3C
34
Step One: To a solution of 33 (88 mg, 0.17 mmol) in ethanol (absolute,
4 mL) at room temperature, NaBH4 (12.5 mg, 0.33 mmol) was added. The
resulting mixture was stirred for 20 minutes, then was quenched with HCI (2N,
2 mL). The resulting mixture was diluted with water and ethyl acetate and the
organic layer was washed with saturated aqueous NaHCO3 and brine. The
organic layer was dried over MgSO, and filtered and the filtrate was
concentrated under reduced pressure to give 35 (85 mg, 96%) as a pale yellow
oil. This material was used without purification.
~ . ~. _.
, .. ,., .~. ..,. ..:.,7
,. .w ,~. ~,_ ,,.. .., ,.;.. _,. ..
CA 02361285 2008-10-23
58
Step Two: To a solution of 35 (85 mg, 0.16 mmol) in ethyl acetate (4
mL) at room temperature under nitrogen, Pd/C (10% dry weight basis, Degussa
type E101 NE/W, -50% water content, 36 nunol) was added. The atmosphere
was replaced with hydrogen (toggle between vacuum and hydrogen from a
balloon five times) and the mixture was vigorously stirred for 1.5 hours. The
mixture was filtered through CeliteTM and the filtrate was concentrated under
reduced pressure. The residue was purified by silica gel chromatography,
eluting with 7:3 hexanes:ethyl acetate to give 36 (32 mg, 39%) as a colorless
oil.
Step Three: To a solution of 36 (32 mg, 0.062 mmol) in THF (3 mL) at
room temperature, aqueous NaOH (2N, I mL) and methanol (2 mL) were added.
The resulting solution was stirred for 15 minutes, diluted with water and
extracted with EtZO. The aqueous phase was acidified with HCl (2N) and was
extracted with ethyl acetate. The ethyl acetate layer was washed with water
and
brine, diied over MgSO4 and filtered. The filtrate was concentrated under
reduced pressure. The residue was taken up in acetonitrile (3 mL) and water (7
mL) and the mixture was lyophilized to give (3S)-3-(1,3-benzodioxol-5-yl)-3-
( {(2S)-2-[2-oxo-3-(phenylmethyl)-1(2H)-pyridinyl]hexanoyl ) amino)propanoic
acid (34, 31 mg, 100%) as a white powder.'H NMR (400 MHz, CD3SOZCD3): 6
0.76 (t, J = 7.3 Hz, 3H), 1.01 (m, 2H), 1.22 (m, 2H), 1.70 (m, IH), 1.87 (m,
IH),
2.60 (dd, J = 15.8, 7.0 Hz, 1 H), 2.68 (dd, J = 15.8, 7.9 Hz, 1 H), 3.72 (d, J
= 15.0
Hz, 1 H), 3.77 (d, J = 15.0 Hz, 1 H), 5.06 (m, 1 H), 5.54 (dd, J= 9.2, 6.6 Hz,
IH),
5.98(s,2H),6.16(t,J=7.0Hz, I H), 6.77 (dd, J = 8.1, 1.4 Hz, 1H),6.83(d,J=
8.1 Hz, 1 H), 6.89 (d, J 1.4 Hz, IH), 7.13 (m, 1 H), 7.18 (m, IH), 7.26 (m,
4H),
7.59 (dd, J = 7.0, 1.8 Hz, 1 H), 8.83 (d, J= 8.1 Hz, 1 H), 12.11 (br. s, 1 H).
Example 13
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-[( { 1-[2-oxo-3-
(phenylmethyl)-1(2H)-pyridinyl]cyclohexyl)carbonyl)amino]propanoic acid.
SteQOne: To a solution of 3-benzylpyridine (1.65 g, 9.77 mmol) in
acetone (3.5 mL), I-chloro-2,4-dinitrobenzene (2.OOg , 9.56 mmol) was added
and the mixture was refluxed ovemight. The mixture was cooled to room.
temperature, diluted with acetone and the solvent was decanted from the
CA 02361285 2001-11-07
59
precipitate. The crude solid was -,vashed with acetone (2 times) and diethyl
ether (1 time), decanting each time to give 37 (3.57 g, 100%) as a gray solid.
Step Two: To a solution of 1-amino-l-hydroxymethylcyclohexane
(0.45g, 3.5 mmol) in n-butanol (8.75 mL), solid N-(2,4-dintrophenyl)-3-
benzylpyridinum chloride (37, 1.23 g, 3.3 nunol) was added. The resulting
solution was heated to reflux for 2.5 days under a nitrogen atmosphere. The
mixture was cooled, diluted with water and filtered. The filtrate was basified
with concentrated NH4OH (2mL) and extracted with ethyl acetate. The aqueous
layer was concentrated to dryness to give 38 (0.56 g) as a yellow oil which
was
used without further purification.
Step Three: To a solution of crude 38 (0.56g, 3.5 mmol theoretical) in
water (10 mL), a solution of potassium ferricyanide (3.3 g, 10 mmol) in water
(15 mL) was added dropwise via an addition fi.tnnel over 30 minutes at 0 C. A
solution of KOH (0.76 g, 13.5 mmol) in water (5 mL) was then added over 30
minutes. Toluene (10 mL) was added and the solution was stirred for one hour
at 0 C. The layers were separated, and the aqueous layer was extracted again
with toluene. The combined extracts were dried over Na,S04and filtered and
the filtrate was concentrated under reduced pressure. The residue was
chromatographed on silica gel, eluting with 7:13 hexanes:ethyl acetate to give
39 (20 mg, 1.9%, two steps.)
Step Four: To a suspension of 39 (20 mg, 0.068 mmol) in aqueous KOH
(1 M, 0.70 mL) potassium persulfate (0.073 g, 0.270 mmol) and ruthenium (III.)
chloride (1mg, catalytic) and THF (0.25 mL) were added. The mixture was
stirred for 1 hour and extracted with dichloromethane. The aqueous layer was
acidified and extracted with ethyl acetate (3 times). The ethyl acetate
extracts
were combined, dried over MgSO4 and filtered. The filtrate was concentrated
under reduced pressure to give 40 (0.0148 g, 70%) as a tan solid.
(3S)-3-(1,3-Benzodioxol-5-yl)-3-[( { 1-[2-oxo-3-(phenylmethyl)-1(2H)-
pyridinyl]cyclohexyl}carbonyl)amino]propanoic acid was prepared from 40
according to the procedures described in Example 1. 'H NMR (400 MHz,
CD3SO2CD,): S 1.40 (m, 4H), 1.68 (m, 2H), 2.04 (m, 2H), 2.60 (d, J = 7.0 Hz,
2H), 3.67 (d, J = 15.2 Hz, 1H), 3.72 (d, J = 15.2 Hz, IH), 5.12 (m, 1H), 5.95
(m,
CA 02361285 2001-11-07
2H), 6.19 (t, J = 7.0 Hz, 1 H), 6.74 (dd, J 7.8, 1.4 Hz, 1 H), 6.76 (d, J =
7.8 Hz,
1H), 6.90 (d, J= 1.4 Hz, 1H), 7.10 (d, J 5.8 Hz, 1H), 7.20 (m, 5H), 7.57 (d, J
= 8.4 Hz, 1 H), 7.66 (dd, J = 7.7, 1.8 Hz, 1 H).
Example 14
5 Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[2-oxo-5-
(phenylmethyl)-1(2H)-pyridinyl]hexanoyl} amino)propanoic acid.
Step One: To a mixture of 41 (prepared according to procedures
described in Example 13, 1.75 g crude orange oil, 5.0 mmol theoretical) in
water
(25 mL) at 0 C, a solution of potassium ferricyanide (4.7 g, 14 mmol) in water
10 (22 mL) was added dropwise via an addition funnel over 30 minutes. A
solution of KOH (l.l g, 19 mmol) in water (7 mL) was then added over 30
minutes. Toluene (15 mL) was added and the solution was stirred for one hour
at 0 C. The layers were separated, and the aqueous layer was extracted again
with toluene. The combined extracts were dried over NazSO4 and filtered and
15 the filtrate was concentrated under reduced pressure. The residue was
chromatographed on silica gel, eluting with 7:13 hexanes:ethyl acetate to give
42 (major product, 0.36 g, 29%) and 43 (minor product, 0.10 g, 7.0%).
(3S)-3-(1,3-Benzodioxol-5-yl)-3-( {2-[2-oxo-5-(phenylmethyl)-1(2H)-
pyridinyl]hexanoyl}amino)propanoic acid was prepared from 42 according to
20 procedures described in Examples I and 13. 'H NMR (400 MHz, CD3SO2CD3)
S: 0.77 (t, J = 7.3 Hz, 3H), 1.00 (m, 2H), 1.20 (m, 2H), 1.75 (m, 1H), 1.88
(m,
1 H), 2.65 (m, 2H), 3.70 (s, 2H), 5.08 (m, IH), 5.49 (dd, J= 9.9, 6.2 Hz, IH),
5.98 (s, 2H), 6.32 (d, J = 9.2 Hz, IH), 6.77 (dd, J = 8.1, 1.5 Hz, 1H), 6.83
(d, J
9.2 Hz, IH), 6.89 (d, J = 1.5 Hz, IH), 7.20 (m, 4H), 7.28 (m, 4H), 7.61 (d, J=
25 2.6 Hz, 1 H), 8.81 (d, J = 8.1 Hz, 1 H), 12.10 (br. s, 1 H).
Example 15
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[(3S)-2,5-dioxo-
3-(phenylmethyl)-4-(2-thiophenylmethyl)tetrahydro-1(2H)-
pyrazinyl]hexanoyl}amino)propanoic acid.
30 Step 1: To a solution of phenylalanine methyl ester (2.32 g, 12.9 mmol)
in DCE (50 ml) at room temperature, 2-thiophenecarboxaldehyde (1.2 ml, 12.9
mmol) and NaBH(OAc)3 (4.11 g, 19.4 mmol) were added. The reaction stirred
at room temperature for 24 hours, diluted with CH2C12 (300 ml) and washed
CA 02361285 2001-11-07
61
with water (300 ml). The organic layer was dried over MgSO4 and filtered and
the filtrate was concentrated under reduced pressure. The residue was purified
by silica gel chromatography, eluting with 9:1 hexanes:ethyl acetate to yield
44
(2.78 g, 78%).
Step 2: To a solution of 44 (1.50 g, 5.45 mmol) in methanol (10 ml),
tetrahydrofuran (10 ml) and water (10 ml), sodium hydroxide (880 mg, 21.8
mmol) was added. The reaction was stirred at room temperature for 48 hours.
The mixture was concentrated under reduced pressure to an aqueous solution
and then lyophilized to yield 45 (1.42 g).
Step 3: To a solution of 45 (500 mg, 1.91 mmol) and norleucine methyl
ester hydrochloride (382 mg, 2.10 mmol) in DMF (10 ml) at room temperature,
1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (401 mg, 2.10
mmol), 1- hydroxybenzotriazole (238 mg, 2.10 mmol) and 4-methylmorpholine
(0.23 ml, 2.10 mmol) were added. The reaction stirred at room temperature for
24 hours then the mixture was taken up in ethyl acetate (200 ml) and washed
with water (2 times, 200 ml). The organic layer was dried over MgSO4 and
filtered and the filtrate was concentrated under reduced pressure. The residue
was purified by silica gel chromatography, eluting with 9:1 hexanes:ethyl
acetate to yield 46 (422 mg, 57%).
Step 4: To a solution of 46 (415 mg, 1.07 mmol) DCE (10 ml) and
triethylamine (0.15 ml, 1.07 mmol) at 0 C, bromoacetyl bromide (0.090 ml,
1.07 mmol) was added and the reaction was warmed to room temperature and
stirred for 24 hours. The mixture was taken up in CH2ClZ (150 ml) and washed
with water (150 ml). The organic layer was dried over MgSO4and filtered and
the filtrate was concentrated under reduced pressure. The residue was purified
by silica gel chromatography, eluting with 4:1 hexanes:ethyl acetate to yield
47
(381 mg, 70%).
Step 5: To a solution of 47 (375 mg, 0.74 mmol) in THF (8 ml), CszCOj
(360 mg, 1.10 mmol) was added. The reaction was stirred at room temperature
for 4 hours. The mixture was taken up in ethyl acetate (150 ml) and washed
with water (150 ml) The organic layer was dried over MgSO., and filtered and
the filtrate was concentrated under reduced pressure. The residue was purified
CA 02361285 2008-10-23
62
by silica gel chromatography, eluting with 4:1 hexanes:ethyl acetate to yield
48
(145.0 mg, 46%).
(3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[(3 S)-2,5-dioxo-3-
(phenylmethyl)-4-(2-thiophenylmethyl)tetrahydro-1(2H)-
pyrazinyl]hexanoyl}amino)propanoic acid was prepared from 48 according to
procedures described in Example 1. MS: Calculated: (M - H)- = 604.2 m/z;
Found: (M - H)- = 604.4 m/z.
Example 16
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[2-oxo-3-
(phenylmethyl)-2,3-dihydro-lH-benzimidazol-l-yl]acetyl}amino)propanoic
acid.
Step One: A mixture of 1-fluoro 2-nitrobenzene (0.50 g, 3.54 mmol),
benzylamine (0.38 g, 3.54 mmol) and K2CO3 (0.98 g, 7.08 mmol) in DMF (10
mL) was stirred at room temperature overnight. The mixture was then
partitioned between EtOAc and water. The organic layer was washed with
water and brine, dried over MgSO4 and filtered. The filtrate was concentrated
to
dryness to give 49 (0.79 g, 98%) as an orange solid.
Step Two: To a solution of 49 (0.79 g, 3.5 mmol) in ethanol (7.0 mL)
and acetic acid (7.0 mL) at room temperature, Fe powder (2.44 g, 34.6 mmol)
was added and the suspension was stirred vigorously at 40 C until thin layer
chromatography indicated complete consumption of 49. The mixture was
filtered through CeliteTM, washing with chloroform. The filtrate was diluted
with
saturated sodium bicarbonate and the chloroform layer was dried over Na2SO4
and filtered. The filtrate was concentrated under reduced pressure and the
residue was purified by chromatography on silica gel (4:1 increasing to 1:1
hexanes:ethyl acetate) to give compound 50 (0.35 g, 50%)
Step Three: A solution of 50 (0.25 g, 1.26 mrnol) and CDI (0.22 g, 1.4
mmol) in CHZC12 (12 mL) was stirred at room temperature overnight. The
mixture was diluted with EtOAc and was washed with 1N HCI (3x) and brine.
The organic layer was dried over MgSO4 and filtered and the filtrate was
concentrated under reduced pressure to give 51 (0.23 g, 82%) as a brown solid.
CA 02361285 2001-11-07
63
Step Four: To a solution of 51 (0.19 g, 0.85 mmol) in anhydrous DMF (5
mL) at 0 C was added NaH (60 % dispersion in mineral oil, 0.044 g, 1.1
mmol). The mixture was stirred at 0 C for 10 minutes before the addition of
ethyl bromoacetate (0.21 g, 0.13 mmol). After stirring at room temperature
overnight, the mixture was poured into ice-water and extracted with EtOAc (2
times). The organic layer was washed with water and brine, dried over MgSO4
and filtered. The filtrate was concentrated under reduced pressure to give 52
(0.25 g, 95 %) as a brown solid.
(3 S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[2-oxo-3-(phenylmethyl)-2,3-
dihydro-lH-benzimidazol-l-yl]acetyl}amino)propanoic acid was prepared from
52 according to procedures described in Example 1. `H NMR (400 MHz,
CD3COCD3): S 2.79 (m, 2H), 4.56 (m, 2H), 5.02 (s, 2H), 5.31 (m, 1H), 5.90 (s,
2H), 6.92 (m, 7H), 7.25 (m, 5H), 7.91 (d, J = 8.4 Hz, 1H), 10.79 (br. S, IH).
Example 17
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[5,5-dimethyl-2,4-
dioxo-3-(phenylmethyl)tetrahydro-1 H-imidazol- l -yl]hexanoyl } amino)
propanoic acid.
Step One: To a solution of 5,5-dimethylhydantoin (2.00 g, 15.6 mmol) in
DMF (30 mL) at room temperature, K,CO; (6.5 g, 47 mmol) and benzyl
chloride (2.20 mL, 18.7 mmol) were added. The resulting mixture was stirred
overnight, diluted with water and extracted with ethyl acetate. The organic
layer
was washed with brine, dried over MgSO4 and filtered. The filtrate was
concentrated under reduced pressure and the residue was purified by silica gel
chromatography, eluting with 6:1 increasing to 3:1 hexanes:ethyl acetate to
yield
53 (3.21 g, 94%).
(3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[5,5-dimethyl-2,4-dioxo-3-
(phenylmethyl)tetrahydro-lH-imidazol-l-yl]hexanoyl}amino)propanoic acid
was prepared from 53 according to procedures described in Examples I and 16.
MP: 53-55 C.
CA 02361285 2001-11-07
64
Example 18
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[2,4-dioxo-l-
(phenylmethyl)-1,4-dihydro-3 (2H)-quinazo linyl ] hexanoyl } amino)propano ic
acid.
Step One: To a solution of anthranillic acid (450 mg, 3.30 mmol) and
norleucine methyl ester hydrochloride (500 mg, 2.75 mmol) in
dimethylformamide (10 ml) at room temperature, 1-[3-(dimethylamino) propyl]-
3-ethylcarbodiimide hydrochloride (640 mg, 3.30 mmol), 1-hydroxybenzo
triazole (450 mg, 3.30 mmol), and 4-methylmorpholine (610 mg, 5.50 mmol)
1.0 were added. The reaction was stirred at room tempc.rature for 24 hours
then the
mixture was taken up in ethyl acetate (200 ml) and washed with water (2 x 200
ml). The organic layer was dried over MgSO4 and filtered and the filtrate was
concentrated under reduced pressure. The residue was purified by silica gel
chromatography, eluting with 4:1 increasing to 1:1 hexanes:ethyl acetate to
yield
54 (860 mg, 99%).
Step Two: A solution of 54 (0.86 g, 3.26 mmol) and CDI (0.79 g, 4.89
mmol) in anhydrous C1CH,CH2C1 (30 mL) was heated at 85 C overnight. The
mixture was cooled to room temperature and concentrated and the residue was
brought up in EtOAc. The organic layer was washed with 1N HC1(3 x) and
brine, dried over MgSO, and filtered. The filtrate was then concentrated under
~ reduced pressure and the residue was purified by silica gel chromatography
( hexanes:EtOAc, 5:1 increasing to 1:1) to give 55 as a white solid (0.67 g,
71%). (3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[2,4-dioxo-l-(phenylmethyl)-
1,4-dihydro-3(2H)-quinazolinyl]hexanoyl}amino)propanoic acid was prepared
from 55 according to procedures outlined in Examples I and 16. 'H NMR (400
MHz, CD3SO2CD3): S 0.80 (m, 3H), 1.1-1.6 (m, 4H), 2.03 (m, 1H), 2.20 (m,
1H),.2.63 (m, 2H), 5.20 (m, 2H), 5.35 (m, 1H), 5.47 (m, 1H), 5.91 (d, J= 11.0
Hz, IH), 5.96 (d, J = 3.3 Hz, 1H), 6.64-6.87 (m, 3H), 7.30 (m, 6H), 7.68 (m,
IH), 8.14 (m, 2H), 12.09 (br. S, IH).
Example 19
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[3-methyl-6-oxo-5-
(phenylmethyl)-1(6H)-pyridazinyl]hexanoyl}amino)propanoic acid.
CA 02361285 2001-11-07
Step One: To a mixture of dihydropyridazinone (2.50 g, 19.21 mmol) in
EtOH (6 mL) at room temperature, benzaldehyde (2.04 g, 19.21 mmol) and
solid KOH (1.3 g, 23.05 mmol) were added. This mixture was heated to 85 C
overnight, cooled to room temperature and poured into ice water. The resulting
5 mixture was acidified to pH = 4 with concentrated HCl and the precipitate
was
collected by filtration, washing with water. The resulting solid was then
dried in
vacuo to give 56 (3.6 g, 85 %).
(3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[3-methyl-6-oxo-5-(phenylmethyl)-
1(6H)-pyridazinyl]hexanoyl}amino)propanoic acid was prepared from 56
10 according to procedures described in Examples 1 and 16. `H NMR (400 MHz,
CD3SO2CD3): S 0.80 (m, 3H), 1.18 (m, 4H), 1.95 (m, 2H), 2.18 (s, 3H), 2:65 (m,
2H), 3.79 (m, 2H), 5.06 (m, 1H), 5.26 (m, 1H), 5.97 (d, J = 2.2 Hz, 2H), 6.81
(m, 3H), 6.98 (s, IH), 7.27 (m, 5H), 8,27 (m, IH), 12.14 (br. S, IH).
15 Example 20
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[2-oxo-3-
(phenylmethyl)-3,4-dihydro-1(2H)-quinazolinyl]hexanoyl} amino)propanoic
acid.
Step 1: A solution of 2-nitrobenzylbromide (0.50 g, 2.31 mmol) and
20 benzylamine (0.49 g, 4.62 mmol) in THF (5 mL) was stirred at room
~ temperature overnight and was then diluted with EtOAc. The organic layer was
washed with 1 N NaOH (twice) and brine, dried over MgSO., and filtered. The
filtrate was concentrated under reduced pressure and the residue was purified
by
silica gel chromatography (hexanes:EtOAc, 3:1 increasing to 1:1) to give 57
25 (0.5 g, 89 %) as an oil.
(3 S)-3-(1,3-benzodioxol-5-yl)-3-( { 2-[2-oxo-3-(phenylmethyl)-3,4-
dihydro-1(2H)-quinazolinyl]hexanoyl}amino)propanoic acid was prepared with
57 according to procedures described in Examples I and 16. `H NMR (400
MHz, CD3SO2CD3): S 0.78 (m, 3H), 1.21 (m, 4H), 1.88 (m, IH), 2.14 (m, 1H),
30 2.65 (m, 2H), 4.26 (m, 2H), 4.43 (m, IH), 4.80 (m, IH), 5.03 (m, 1 H), 5.18
(m,
1 H), 5.92 (d, J= 4.0 Hz, IH), 5.97 (s, 1 H), 6.70 (m, 2H), 6.80 (m, 1 H),
6.95 (m,
2H), 7.12 (m, 2H), 7.30 (m, 5H), 8.20 (m, IH), 12.09 (br. s, IH).
CA 02361285 2001-11-07
66
Example 21
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[2-oxo-3-
(phenylmethyl)-1(2H)-quinoxalinyl]hexanoyl} amino)propanoic acid.
Step One: To solution of 1,2-phenylenediamine (2.64 g, 14.4 mmol) and
phenylpyruvic acid (2.00 g, 12.2 mmol) in ethanol (absolute, 20 mL), a
solution
of 2-mercaptoethanol (1.6 mL) in 2N HCl (18.3 mL) was added. The resulting
mixture was heated to reflux for 2 hours, then was allowed to cool to room
temperature and filtered, washing the precipitate with ethanol (twice). The
precipitate was dried under vacuum to give 58 (1.88 g, 65%) as a white solid.
(3 S)-3-(1,3-Benzodioxol-5-yl)-3-( {2-[2-oxo-3-(phenylmethyl)-1(2H)-
quinoxalinyl)hexanoyl}amino)propanoic acid was prepared from 58 according
to procedures described in Examples I and 16. MS: Calculated (M - H)- _
540.21; Found (M - H)- = 540.21.
Example 22
Synthesis of(3S)-3-(1,3-benzodioxol-5-yl-3-((2S)-2-(3-benzyl-5-methyl-
2-oxo-1(2H)-pyridinyl)hexanoylamino)propanoic acid, (10).
Step One: To a solution of (R)-(+)-2-bromohexanoic acid (410 mg, 2.1
mmol) and NMM (0.265 mL, 2.1 mmol) in THF ( 8 mL) at 0 C under a dry,
nitrogen atmosphere, isobutyl chloroformate (0.27 mL, 2.1 mmol) was added.
The resulting suspension was stirred at 0 C for 10 minutes before the addition
of a solution of 6 (500 mg, 2.1 nunol) in DMF (2 mL). After I hour the
suspension was filtered and the filtrate was concentrated under reduced
pressure. The residue was taken up in CHzCIz and washed with HCI (1 N, 2
times). The combined organic layers were washed with brine. The organic
layer was dried over MgSO4, filtered and concentrated under reduced pressure
to
give 59 (840 mg, 96% yield) as a yellow oil. This material was used without
further purification.
Step Two: To a solution of 2-fluoro-5-methylpyridine (3.23 g, 30 mmol)
in THF (60 mL) cooled to -78 C under a dry, nitrogen atmosphere, lithium
diisopropylamide (22 mL, 2 M solution in THF, 44 mmol) was added dropwise.
The resulting orange-red solution was stirrPd at -78 C for 6 hours after
which
benzaldehyde (3.1 mL, 35 mmol) was added. The reaction mixture was stirred
CA 02361285 2001-11-07
67
an additiona130 minutes, quenched with H20 (75 mL) and extracted with
EtOAc (2 times). The combined organic layers were washed with brine, dried
over MgSO4, filtered and concentrated under reduced pressure to give 60 (6.5
g,
quant.) as a yellow oil. This material was used without further purification.
Step Three: A solution of 60 (6.5 g, 30 mmol), boron trifluoride diethyl
etherate (13 mL, 100 mmol) and triethylsilane (9 mL, 60 mmol) in
dichloroethane (50 mL) was heated to reflux under a dry, nitrogen atmosphere
and for 1 hour. The resulting solution was allowed to cool to room temperature
and quenched with HZO (50 mL). The resulting mixture was extracted with
EtOAc (2 times). The combined organic layers were washed with brine, dried
over MgSO4, filtered and concentrated under reduced pressure to give 61 (5.9
g,
98 % yield) as a yellow oil.
Step Four: A solution of 61 (5.9 g, 30 mmol) in dioxane (2 mL) was
added to HCI (6 N, 50 mL) and the resulting mixture was heated to reflux for
12
hours. The resulting solution was allowed to cool to room temperature and
made basic by the addition of NaOH (2 N). The resulting mixture was extracted
with EtOAc (100 mL x 2) and the combined organic layers were washed with
brine, dried over MgSO4 and filtered. The resulting filtrate was concentrated
under reduced pressure. The residue was recrystallized from methanol/hexanes
to provide 62 (3.1 g, 51% yield) as an off-white solid.
Step Five: To a solution of 62 (48 mg, 0.24 mmol) in DME (1.5 mL)
under a dry, nitrogen atmosphere cooled to -40 C, NaHMDS (0.25 mL, 1M in
THF, 0.25 mmol) was added. The resulting suspension was stirred at -40 C for
an additional 30 minutes before LiBr (21 mg, 0.25 mmol) was added. After an
additional 10 minutes, a solution of 59 (100 mg, 0.24 mmol) in DMF (0.5 mL)
was added dropwise. The resulting mixture was then allowed to warm to room
temperature and then was heated to 45 C for 3.5 hours. The mixture was
allowed to cool to room temperature and quenched with HCI (1 N, 15 mL). The
resulting mixture was extracted with CH2C12 (twice). The combined organic
layers were washed with aqueous sodium carbonate (5%) and brine. The organic
phase was dried over MgSO4filtered and concentrated under reduced pressure.
CA 02361285 2001-11-07
68
The residue was recrystallized from methanoUhexanes to give 63 (39 mg, 31 %)
as white crystals.
(3 S)-3-(1,3-benzodioxol-5-yl-3-((2S)-2-(3-benzyl-5-methyl-2-oxo-
1(2H)-pyridinyl)hexanoylamino)propanoic acid, (10),was prepared from 63
according to procedures described in Example 1.
Among the compounds representative of the present invention, which
can be synthesized according to the procedure of Example 22, by varying
starting materials, are (3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-
yl)hexanoyl]amino}-3-(3',4'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)-3-(3',4'-dimethoxy-l,l'-biphenyl-4-yl)propanoic acid, (3S)-
3-(1,1'-biphenyl-4-yl)-3-( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl} amino)propanoic acid, (3S)-3- {[(2S)-2-(3-
benzyl-5-methyl-2-oxopyri din-1(2H)-yl)hexanoyl] amino) -3-(1,1'-biphenyl-4-
yl)propanoic acid, (3S)-3-(1,1'-biphenyl-4-yl)-3-({(2S)-2-[3-(2-chlorobenzyl)-
5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)propanoic acid, (3S)-3-{[(2S)-
2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl] amino} -3-(4'-methyl-
1,1'-biphenyl-4-yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(4'-methyl-1,1'-biphenyl-4-
yl)propanoic acid, (3S)-3-({(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-
~~ oxopyridin-1(2H)-yl]hexanoyl}amino)-3-(4'-methyl-1,1'-biphenyl-4-
yl)propanoic acid, (3S)-3- {[(2S)-2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-
yl)hexanoyl]amino}-3-(2',6'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)-3-(2',6'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3-( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)-3-(2',6'-dimethoxy-1,1'-biphenyl-4-yl)propanoic acid, (3S)-
3- {[(2S)-2-(3-benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyl]amino}-3-(1,1'-
biphenyl-3-yl)propanoic acid, (3S)-3-(l, l'-biphenyl-3-yl)-3-( {(2S)-2-[3-(2-
chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl}amino)propanoic acid,
(3S)-3-(1,1'-biphenyl-3-yl)-3-( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-
2-oxopyridin-1(2H)-yl]hexanoyl}amino)propanoic acid, (3S)-3-{[(2S)-2-(3-
benzyl-5-methyl-2-oxopyridin-1(2H)-yl)hexanoyljamino } -3-(2'-methoxy-1,1'-
CA 02361285 2001-11-07
69
biphenyl-3-yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(2'-methoxy-1,1'-biphenyl-3-
yl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chloro-6-methylbenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(2'-methoxy-1,1'-biphenyl-3-
yl)propanoic acid, (3S)-3-({(2S)-2-[3-(2-fluoro-6-methoxybenzyl)-5-methyl-2-
oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(4-methylphenyl)propanoic acid, (3S)-
3-( {(2S)-2-[3-(2,6-dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-3-( {(2S)-2-[3-(3,5-
dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(4-
methylphenyl)propanoic acid, (3S)-3-({(2S)-2-[3-(2,6-dichlorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(4-methylphenyl)propanoic
acid, (3S)-3-( {(2S)-2-[3-(2-chloro-6-fluorobenzyl)-5-methyl-2-oxopyridin-
1(2H)-yl]hexanoyl}amino)-3-(4-methylphenyl)propanoic acid, (3S)-3-({(2S)-2-
[3-(2,6-difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(4-
methylphenyl)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[3-(2,6-
difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)propanoic
acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[3-(2-chloro-6-fluorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl}amino)propanoic acid, (3S)-3-(1,3-
benzodioxol-5-yl)-3-( {(2S)-2-[3-(2,6-dichlorobenzyl)-5-methyl-2-oxopyridin-
1 (2H)-yl]hexanoyl } amino)propanoic acid, (3 S)-3-(1,3-benzodioxol-5-yl)-3-
~ ( {(2S)-2-[3-(2,6-dimethylbenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl} amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yi)-3-({(2S)-2-
[ 3-(3, 5 -dimethylbenzyl)-5-methyl-2-oxopyri din-1(2H)-
yl]hexanoyl} amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-
[3-(2-fluoro-6-methoxybenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl} amino)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-
methyl-2-oxopyridin-1(2H)-yl]hexanoyl } amino)-3-(3,4-
diethoxyphenyl)propanoic acid, (3S)-3-(3,4-diethoxyphenyl)-3-( {(2S)-2-[3-(2,6-
difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)propanoic
acid, (3S)-3-( {(2S)-2-[3-(2,6-difluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-
yl]hexanoyl} amino)-3-(3-ethoxyphenyl)propanoic acid, (3S)-3-( {(2S)-2-[3-(2-
chloro-6-fluorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]hexanoyl} amino)-3-(3-
ethoxyphenyl)propanoic acid(3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-
CA 02361285 2001-11-07
oxopyridin-1(2H)-yl]-3-methylbutanoyl} amino)-3-(4-methylphenyl)propanoic
acid, (3S)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-i (2H)-yl]-4-
methylpentanoyl} amino)-3-(4-methylphenyl)propanoic acid, (3 S)-3-(1,3-
benzodioxol-5-yl)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-
5 1(2H)-yl]-4-methylpentanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-
yl)-3-( {(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyri din-1(2H)-yl]-3-
methylbutanoyl}amino)propanoic acid, (3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-
2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-1(2H)-yl]-3-
phenylpropanoyl } amino)propanoic acid, (3 S)-3-(1,3-benzodioxol-5-yl)-3-
10 (((2R)-2-(1-benzyl-5-methyl-2-oxo-1, 2-dihydropyridin-3 -yl)hexanoyl)amino)
propanoic acid and (3S)-3-({(2S)-2-[3-(2-chlorobenzyl)-5-methyl-2-oxopyridin-
1(2H)-yl]-3-phenylpropanoyl} amino)-3-(4-methylphenyl)propanoic acid.
Example 23
Synthesis of (3S)-3-(1,3-benzodioxol-5-yl)-3- {[(2R)-2-(3-benzyl-4-
15 hydroxy- 5 -methyl-2 -oxopyri din-1(2H)-yl)hexanoyl]amino}propanoic acid,
67,
and (3S)-3-(1,3-benzodioxol-5-yl)-3-{[(2S)-2-(3-benzyl-4-hydroxy-5-methyl-2-
oxopyridin-1(2H)-yl)hexanoyl]amino}propanoic acid, 68.
Step One: To a solution of methyl propionylacetate (2.OOg 15.4 mmol) in
anhydrous THF (62 mL) cooled to 0 C under a dry nitrogen atmosphere,
20 sodium hydride (60% dispersion in mineral oil, 0.644g, 16.1 mmol) was added
in two portions. The resulting mixture was stirred for 15 minutes at 0 C,
benzyl bromide (2.75g, 16.1 mmol) was added by syringe then the mixture was
allowed to warm to room temperature overnight. The resulting mixture was
poured into HCI (2N) and extracted with ethyl acetate. The organic layer was
25 washed with brine, dried over Mg SO., and filtered and the filtrate was
concentrated under reduced pressure. The residue was purified by silica gel
chromatography, eluting with 9:1 hexanes:ethyl acetate to give 64 (2.86 g, 84%
yield).
Step Two: To a suspension of sodium hydride (60% dispersion in mineral oil,
30 0.623g, 15.6 mmol) in THF (52 mL) at room temperature under a dry nitrogen
atmosphere, N,N,N',N'-tetramethylethylenediamine (2.10 mL, 13.6 mmol) was
added. The resulting suspension was cooled to -20 C and a solution of 64
CA 02361285 2001-11-07
71
(2.86g, 13.0 mmol) in THF (5 mL) was added slowly. The mixture was allowed
to warm to room temperature, stirred for 10 minutes then was cooled to -20 C.
To the resulting solution, n-butyllithium (1.6M in hexanes, 15.4 mL, 24.9
mmol) was added dropwise by syringe. The mixture was stirred at -20 C for an
additional 15 minutes, then quenched with the rapid addition of methyl formate
(1.00 mL, 16.2 mmol) via syringe and the mixture was allowed to stir for an
additional 15 minutes. The reaction was cautiously quenched with excess
aqueous HCl (2N) and diluted with hexanes. The hexanes layer is washed with
brine, dried over MgSO4, filtered and concentrated under reduced pressure to
give 65 (1.10g, 34%) as a yellow oil. This material was used without further
purification.
Step Three: A solution of 65 (0.75g, 3.0 mmol) and 23 (0.75 g, 2.1 mmol) in
methanol (30 mL, 0.1M) was heated at 45 'C under nitrogen for 1 hour before
refluxing overnight. The mixture was cooled and concentrated to dryness. The
residue was purified by silica gel chromatography, eluting with 2:1
hexanes/ethyl acetate, increasing to ethyl acetate to give 66 (0.40 g, 35%
yield)
as a white foam.
Step Four: To a solution of 66 (0.40 g, 0.73 mmol) in THF (3 mL) at room
temperature, aqueous NaOH (1 mL) and methanol (2 mL) were added. The
resulting solution was stirred for 15 minutes, diluted with water and
extracted
with ethyl ether. The aqueous layer was acidified with HCl (2N) and extracted
with ethyl acetate. The ethyl acetate layer was washed with water and brine,
dried over MgSO4 and filtered. The filtrate was concentrated under reduced
pressure and the residue was purified by reverse phase HPLC (A: 19:1
H20:CH'CN + 0.1 % TFA; B:19:1 CH3CN:H20 + 0.1 %TFA; gradient elution
30% B to 100% B in 31 minutes; 254 nM). The fractions containing each of the
diastereomers were separately diluted with water and extracted with ethyl
acetate. The ethyl acetate layers were separately washed with water (3 times)
and brine, dried over MgSO, and filtered and the filtrates were separately
concentrated under reduced pressure to give (3S)-3-(1,3-benzodioxol-5-yl)-3-
{ [(2R)-2-(3-benzyl-4-hydroxy-5-methyl-2-oxopyridin-1(2H)-
yl)hexanoyl]amino}propanoic acid, 67, (18 mg, 5% yield) and (3S)-3-(1,3-
CA 02361285 2001-11-07
72
benzodioxol-5-yl)-3 - {[(2S)-2-(3-benzyl-4-hydroxy-5-methyl-2-oxopyridin-
1(2H)-yl)hexanoyl]amino}propanoic acid, 68, (105 mg, 28% yield). MS for 67:
Calculated (M+H)+ = 521.23; Found (M+H)+ = 520.97. MS for 68: Calculated
(M+H)+ = 521.23; Found (M+H)+ = 520.95.
Example 24
Synthesis of (3S)-3-(3-isopropoxyphenyl)-3-( {2-[2-oxo-4-(2-
phenylethyl)pyridin-1(2H)-yl]hexanoyl } amino)propanoic acid.
Step One: To a solution of diisopropylamine (2.94 g, 29.1 mmol) in THF
(20 mL) cooled to -78 C under a dry nitrogen atmosphere, butyllithium (12.8
mL of a 2.5 M solution in hexanes, 32.0 nunol) was added by syringe and
stirred
at -78 C for 15 minutes. The solution was added by cannula to a solution of 2-
fluoro-4-methyl-pyridine (2.15 g, 9.40 mmol) in THF (20 mL) cooled to -78 C
under a dry nitrogen atmosphere. The resulting mixture was stirred at -78 C
for
4 hours, and then quenched with benzaldehyde (2.17 mL, 21.3 mmol). The
resulting yellow solution was warmed to room temperature, diluted with water
and extracted with ethyl acetate. The organic layer was washed with brine,
dried
over MgSO.4 and filtered. The filtrate was concentrated under reduced pressure
and the residue was purified by silica gel chromatography, eluting with 7:3
hexanes:ethyl acetate to give 69 (1.92 g, 94%) as a yellow oil.
Step Two: To a solution of 69 (1.92 g, 8.84 mmol) in anhydrous methanol (20
mL) at room temperature under a dry nitrogen atmosphere, palladium on
charcoal (1.00 g, 10% Pd dry weight basis, Degussa type E l Ol NE/W, wet, 50%
water by weight) and four drops of glacial acekc acid were added. The nitrogen
atmosphere was replaced by hydrogen (alternate five times between vacuum and
hydrogen supplied by balloon) and the mixture was stirred at room temperature
ovemight. The mixture was filtered through Celite 521 and the filtrate was
concentrated under reduced pressure. The residue was purified by silica gel
chromatography, eluting with 8:2 hexanes:ethyl acetate to give 70 (0.98 g,
55%)
as a white solid.
Step Three: To a solution of 70 (0.90 g, 4.47 mmol) in hydrogen chloride (2
mL,
4.0 M in dioxane, 8.0 mmol) at room temperature, aqueous hydrogen chloride
(6.0 M, 40 mL) was added and the mixture was refluxed for 3 hours. The
CA 02361285 2001-11-07
73
solution was cooled to room temperature, made basic with aqueous sodium
hydroxide (2N), and extracted with ethyl acetate. The organic layer was washed
with brine, dried over MgSO4 and filtered. The filtrate was concentrated under
reduced pressure to give 71 (0.81 g, 91 %) as a pale yellow solid. This
material
was used without purification
Step Four: To a solution of 71 (0.20 g, 1.0 mmol) in methyl sulfoxide (4.0 mL)
at room temperature, sodium hydride (60% w/w dispersion in mineral oil, 0.12
g, 3.0 mmol) was added. The mixture was stirred at room temperature under a
dry nitrogen atmosphere for 1 hour. To the resulting mixture, ethyl 2-
bromohexanoate (0.18 mL, 1.0 mmol) was added and the mixture was stirred
for 30 minutes. The mixture was quenched with HCl (2N, 50 mL) and extracted
with ethyl acetate. The organic layer was washed with brine, dried over MgSO4
and filtered. The filtrate was concentrated under reduced pressure and the
residue was purified by silica gel chromatography, eluting with 9:1
hexanes:ethyl acetate, increasing to 8:2 hexanes:ethyl acetate to give 72
(0.19 g,
0.55 mmol) as a solid.
(3S)-3-(3-isopropoxyphenyl)-3-( {2-[2-oxo-4-(2-phenylethyl)pyridin-
1(2H)-yl]hexanoyl}amino)propanoic acid was prepared from 72 according to
procedures described in Example 1. MS: Calculated (M-H)- = 517.27; Found
(M-H)- = 517.21.
Example 25
Synthesis of (3S)-3-[3-(difluoromethyl)phenyl]-3-( {2-[5-methyl-2-oxo-
3-(phenylmethyl)-1(2H)-pyridinyl]hexanoyl}amino)propanoic acid.
Step One: To a solution of 3-bromobenzaldehyde, 73, (3.00 g, 16.2
mmol) in DMF (69 mL) under a dry nitrogen atmosphere, palladium acetate (73
mg, 0.32 mmol), tri-o-tolylphosphine (197 mg, 0.65 mmol), ethyl acrylate (2.20
mL, 20.3 mmol) and triethylamine (4.50 mL, 32.4 mmol) were added. The
system was deoxygenated (toggle between vacuum and nitrogen five times), the
mixture was heated to 125 C for 19 hours and then cooled to room temperature.
The reaction was poured into water and extracted with ether. The organic layer
was washed with HCl (4N) and brine, dried over MgSO4and filtered. The
CA 02361285 2008-10-23
74
filtrate was concentrated under reduced pressure to give 74 (2.74g, 83%),
which
was used without further purification.
Step Two: To a flask containing 74 (1.00 g, 4.9 mmol) under a dry
nitrogen atmosphere, (dimethylamino)sulfur trifluoride (0.96 mL, 9.8 mmol)
was added by syringe. The mixture was heated to 90 C behind a blast shield
for
25 minutes then was cooled to room temperature. The resulting mixture was
diluted with CHZCIz and washed with saturated aqueous NaHCO3 and H20. The
organic layer was dried over MgSO4 and filtered and the filtrate was
concentrated under reduced pressure. The residue was purified by silica gel
chromatography, eluting with 1:5 ethyl actetate:hexanes to give 75 (0.62 g,
56%).
Step Three: To a solution of (R)-(+)-N-benzyl-a-methylbenzylamine (0.70 g,
3.3 mmol) in THF (6.7 mL) cooled to -78 C under a dry nitrogen atmosphere,
sec-BuLi (4.22 mL, 1.3M in cyclohexane, 5.5 mmol) was added dropwise. The
resulting mixture was stirred at -78 `C for 30 minutes and then a solution of
75
(0.62 g, 2.74 mmol) in THF (3.4 mL) was added dropwise by syringe. The
mixture was stirred at -78 C for 5 hours and then quenched with glacial AcOH
(2 mL) in THF (5 mL). The reaction mixture was warmed to room temperature,
poured into a 1:1 mixture of saturated aqueous NaHCO3:EtOAc. The organic
layer was washed with H20 (2 times) and brine, dried over MgSO4 and filtered.
The filtrate was concentrated under reduced pressure and the residue was
purified by silica gel chromatography, eluting with 1:5 ethyl actetate:hexanes
to
give 76 (1.2 g, 100%). This material still contained minor impurities but was
used without further purification.
Step Four: To a solution of 76 (0.50 g, 1.14 mmol) in EtOH (10 mL) at
room temperature under a dry nitrogen atmosphere, Pd/C (10% Pd dry weight
basis, 50% water by weight, Degussa type E 101 NE/W, 0.25 g) and glacial
AcOH (0.5 mL) were added. The atmosphere was replaced by hydrogen (toggle
between vacuum and hydrogen from a balloon five times) and the mixture was
heated to 35 C for 6 hours. The reaction was cooled to room temperature,
filtered through a plug of CeliteTM 521 and the filtrate was concentrated
under
reduced pressure. The residue was diluted with CHCl3 and washed with
CA 02361285 2001-11-07
saturated aqueous NaHCO3. The aqueous layer was extracted with CHC13 (2
times) and the combined organic layers were dried over MgSO4 and filtered.
The filtrate was concentrated under reduced pressure and the residue was
purified by silica gel chromatography, eluting with 1:10 MeOH:CHCl3 to give
5 77 (180 mg, 67%).
(03S)-3-[3-(difluoromethyl)phenyl]-3-( {2-[5-methyl-2-oxo-3-
(phenylmethyl)-1(2H)-pyridinyl]hexanoyl } amino)propanoic acid was
synthesized from 77 according to procedures described ir. Example 1. MS:
Calculated (M-H)- = 509.23; Found (M-H)- = 509.19.
10 Example 26
The ability of compounds of the present invention to inhibit binding is
determined by a procedure in which a 26-amino acid peptide containing the CS I
sequence of fibronectin with an N-terminal Cys
(CDELPQLVTLPHPNLHGPEILDVPST) was coupled to maleimide activated
15 ovalbumin. Bovine serum albumin (BSA) and CS 1 conjugated ovalbumin were
coated onto 96-well polystyrene plates at 0.5 mg/ml in TBS (50 mM Tris, pH
7.5; 150 mM NaCI) at 4'C for 16 hours. The plates were washed three times
with TBS and blocked with TBS containing 3% BSA at room temperature for 4
hours. Blocked plates were washed three times in binding buffer (TBS; 1 mM
20 MgCl2; 1 mM CaC12; 1 mM MnCI2) prior to assay. Ramos cells fluorescently
labeled with calcein AM were resuspended in binding buffer (107 cells/ml) and
diluted 1:2 with same buffer with or without compound. The cells were added
immediately to the wells (2.5 x 105 cells/well) and incubated for 30 minutes
at
37 C. Following three washes with binding buffer, adherent cells were lysed
25 and quantitated using a fluorometer. The results are shown in Tables 1, 2,
3 and
4. IC50 is defined as the dose required to give 50% inhibition. MS in Table 3
stands for Mass Spec. nd stands for not determined in the Tables. A stands for
inhibition in Table 2, and the percent inhibition indicates the inhibition of
cell
adhesion when compound is included in the assay at a concentration of 100 M.
30 The lower the ICso value and the greater the percentage of inhibition, the
more
efficient the compound is at prevention of cell adhesion.
CA 02361285 2001-11-07
76
Table 1
Compound Number IC50 (nM) Mass Spectral Data (m/z)
8 7 Calc'd (M-H)" = 503.22
Found (M-H)- = 503.24
9 2000 Calc'd (M-H)- = 503.22
Found (M-H)- = 503.24
10 3 Calc'd (M-H)- = 503.22
Found (M-H)- = 503.22
12 4(, Calc'd (M-H)- = 501.24
Found (M-H)- = 501.27
13 70 Calc'd (M-H)- = 473.24
Found (M-H)- = 473.26
14 150 Calc'd (M-H)- = 477.22
Found (M-H)- = 477.25
30 Calc'd (M+H)` = 535.25
Found (M+H)+ = 535.00
10 16 ~5 Calc'd (M+H)' = 519.25
Found (M+H) = 519.18
17 60 Calc'd (M-H)- = 521.21
Found (M-H)- = 521.22
19 6 Calc'd (M-H) = 537.18
Found (M-H)- = 537.22
CA 02361285 2001-11-07
77
Table 2
Compound IC50 % A Mass Spectral Data
( M) mJz
(3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2- 0.15 100 Calc'd (M-H)- =
[2-oxo-3-(2-thienylmethyl)-1- 486.37
imidazolidinyl]hexanoyl } amino)propanoic Found (M-H)- =
acid 486.20
(3S)-3-(1,3-benzodioxol-5-yl)-3-(((2S)-2- 0.005 100 Calc'd (M-H)- =
(2-oxo-3-(2-thienylmethyl)tetrahydro- 500.18
l(2H)- Found (M-H)- =
pyrimidinyl)hexanoyl)amino)propanoic 500.22
acid
(3S)-3-(1,3-benzodioxol-5-yl)-3-(((2S)-2- 0.05 100 Calc'd (M-H)- =
(2-oxo-4-(2-thienyl)-3-(2- 582.17
thienylmethyl)tetrahydro-1(2H)- Found (M-H)- =
pyrimidinyl)hexanoyl)amino)propanoic 582.21
acid
(3S)-3-(1,3-benzodioxol-5-y1)-3-({(2S)-2- nd nd nd
[2-oxo-3-(2-thienylmethyl)-1,3-diazepan-
1- 1 hexano 1 amino ro anoic acid
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78
Table 3
Compound Mass Spectral Data IC50
(m/z) M)
(3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[2,5- Calc'd (M-H)- = 1.5
dioxo-1-(2-thiophenylmethyl)-1,2,3,5- 576.18
tetrahydro-4H- 1,4-benzodiazepin-4- Found (M-H)" =
1 hexano 1}amino ro anoic acid 576.18
(3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2- Calc'd (M-H)- = 6
[(3S)-2,5-dioxo-3-(phenylmethyl)-4-(2- 604.21
thiophenylmethyl)tetrahydro- 1(2H)- Found (M-H)- =
pyrazinyljhexanoyl)aniino)propanoic acid 604.24
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[3- Calc'd (M-H)- = >100
(phenyloxy)phenyl]acetyl } amino)propanoic 418.11
acid Found (M-H)- =
418.12
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 1.3
[(2-thiophenylmethyl)amino]-1(2H)- 510.17
pyridinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
510.21
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 0.2
(phenylmethyl)-2,3-dihydro-1 H-benzinlidazol- 472.15
1-yl]acetyl}amino)propanoic acid Found (M-H)- =
472.18
(3S)-3-(1,3-benzodioxol-5-yl)-3-( ',2-[2-oxo-3- Calc'd (M-H)- = 0.07
(phenylmethyl)-2,3-dihydro-1 H-benzimidazol- 528.21
1-yl]hexanoyl}amino)p.opanoic acid Found (M-H)- =
528.22
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 0.03
(phenylmethyl)-1(2H)- 461.17
pyridinyl]butanoyl}amino)propanoic acid Found (M-H) =
461.18
(3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[2- Calc'd (M-H)- = 0.55
oxo-3-(phenylcarbonyl)-1(2H)- 503.18
pyridinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
503.18
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 0.45
(phenylmethyl)-1(2H)- 433.14
pyridinyl]acetyl}amino)propanoic acid Found (M-H)- =
433.16
(3S)-3-(1,3-benzodioxol-5-yl)-3-[( { 1-[2-oxo-3- Calc'd (M-H)- = 50
(phenylmethyl)-1(2H)- 501.20
pyridinyl]cyclohexyl}carbonyl)amino] Found (M-H)- =
propanoic acid 501.24
(3S)-3-(1,3-benzodioxol-5-yl)-3-( {(2S)-2-[2- Calc'd (M-H)- = 0.004
oxo-3-(phenylmethyl)-1(2H)- 489.20
pyridinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
489.20
CA 02361285 2001-11-07
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(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 0.06
(phenylmethyl)-2,3-dihydro-lH-imidazol-l- 478.20
yl]hexanoyl}amino)propanoic acid Found (M-H)- =
478.23
(3S)-3-(1,3-benzodioxol-5-yl)-3-({(2S)-2-[2,4- Calc'd (M-H)- = 0.1
dioxo-l-(phenylmethyl)-1,4-dihydro-3(2H)- 556.21
quinazolinyl]hexanoyl} amino)propanoic acid Found (M-H)- =
556.22
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[3-[(2- Calc'd (M-Hy = 0.01
chlorophenyl)methyl]-5-methyl-2-oxo-1(2H)- 537.18
pyridinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
537.22
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[5,5- Calc'd (M-H)- = 20
dimethyl-2,4-dioxo-3- 522.22
(phenylmethyl)tetrahydro-1 H-imidazol-l- Found (M-H)- =
I hexano 1 amino ro anoic acid 522.22
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-5- Calc'd (M-H)- = 0.04
(phenylmethyl)-1(2H)- 489.20
pyridinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
489.21
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 100
(phenylmethyl)-1(2H)- 540.21
quinoxalinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
540.21
(3S)-3-(1,3-benzodioxol-~-yl)-3-( {2-[2-oxo-3- Calc'd (M-H)- = 0.06
(phenylmethyl)-1(2H)- 475.18
pYndinY1]pentanoY1}amino)proPanoic acid Found (M-H)- =
475.19
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[3-methyl- Calc'd (M-H)- = 0.06
6-oxo-5-(phenylmethyl)-1(6H)- 504.21
pyridazinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
504.24
(3S)-3-(1,3-benzodioxol-5-yl)-3-({2-[2-oxo-3- Calc'd (M-H)- = 0.4
(phenylmethyl)-3,4-dihydro-1(2H)- 542.23
quinazolinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
542.26
(3S)-3-(1,3-benzodioxol-5-yl)-3-( {2-[2-oxo-3- Calc'd (M-H)- = 2
(phenylmethyl)-1(2H)- 539.22
quinolinyl]hexanoyl}amino)propanoic acid Found (M-H)- =
539.22
CA 02361285 2001-11-07
Table 4
Compound IC50 Mass Spectral Data
(nM) (m/z)
3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2- 10 easured (M-H)- = 519.25;
xopyridin-1(2H)-yl)hexanoyl]amino}-3- alculated (M-H)- = 519.25.
5 3,5-dimethoxyphenyl)propanoic acid
3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2- 17 easured (M-H)- = 507.13;
xopyridin-1(2H)-yl)hexanoyl]amino}-3-(3- alculated (M-H)- = 507.23.
uoro-4-methoxyphenyl)propanoic acid
3S)-3-(1,3-benzodioxol-5-yl)-3-{[2-(3- 30 easured (M-H) = 489.17;
10 enzyl-2-oxopyridin-1(2H)- alculated (M-H)- = 489.20.
1)hexanoyl]amino}propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2- 10 easured (M-H) = 517.19;
xopyridin-1(2H)-yl)hexanoyl]amino}-3-(3- alculated (M-H)- = 517.27.
wf
ropoxyphenyl)propanoic acid
15 3S)-3-{[2-(3-benzyl-2-oxopyridin-1(2H)- 300 easured (M+H)+ = 504.94;
1)hexanoyl]amino}-3-(4-methoxy-2,3- alculated (M+H)+=
imethylphenyl)propanoic acid 505.27.
3S)-3-{[(2S)-2-(3-benzyl-5-methyl-2- 200 easured (M-H)- = 595.18;
xopyridin-1(2H)-yl)hexanoyl]amino} -3- Calculated (M-H)- = 595.28.j
20 2',6'-dimethoxy-1,1'-biphenyl-4-
l)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2- 130 Measured (M-H)- = 487.24;
xopyridin-1(2H)-yl)hexanoyl]amino}-3- alculated (M-H) =487.26.
3,4-dimethylphenyl)propanoic acid
25 3S)-3-{[2-(3-benzyl-5-methyl-2- 35 4easured (M+H)+ = 504.99;
~., xopyridin-1(2H)-yl)hexanoyl]amino}-3-(3- alculated (M+H)` _
thoxyphenyl)propanoic acid 505.27.
3S)-3-{[2-(3-benzyl-5-methyl-2- 33 easured (M+H)+ = 550.93;
xopyridin-1(2H)-yl)hexanoyl]amino } -3- alculated (M+H)+
30 3,4,5-trimethoxyphenyl)propanoic acid 551.28.
3S)-3-{[2-(3-benzyl-5-methyl-2- 140 easured (M+H)+ = 518.98;
xopyridin-1(2H)-yl)hexanoyl]amino } -3-(4- alculated (M+H)+ _
ethoxv-3,5-dimethylphenyl)propanoic acid~ 519.29.
3S)-3-{[2-(3-benzyl-5-methyl-2- 40 1Meased (M+H) = 540.98;
35 xopyridin-1(2H)-yl)hexanoyl]amino}-3-(6- ~alculated (M+H)' _
ethoxy-2-naphthyl)propanoic acid 41.27.
3S)-3-{[2-(3-benzyl-5-methyl-2- 45 1easured (M+H)` = 533.03;
xopyridin-1(2H)-y1)hexanoyl]amino} -3-(3- ~alculated (M+H)- _
utoxyphenyl)propanoic acid -33.30.
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3S)-3-(1,3-benzodioxol-5-yl)-3-{[(2S)-2- 200 easured (M+H)` = 520.97=
3-benzyl-4-hydroxy-5-methyl-2- alculated (M+H)+ _
xopyridin-1(2H)- 521.23.
1)hexanoyl]amino}propanoic acid
3S)-3-(1,3-benzodioxol-5-yl)-3-{[(2R)-2- 2000 easured (M+H)+ = 520.95;
3-benzyl-4-hydroxy-5-methyl-2- alculated (M+H)+
_
xopyridin-1(2H)- 521.23.
1)hexanoyl]amino}propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2- 4 easured (M+H)+ = 518.99;
xopyridin-1(2H)-yl)hexanoyl]amino } -3-(3- alculated (M+H)+ =
sopropoxyphenyl)propanoic acid 519.29.
3S)-3-({2-[3-(2,6-dichlorobenzyl)-5-methyl 275 alculated (M+H)- =
-oxopyridin-1(2H)-yl]hexanoyl}amino)-3-(3 587.21; Found (M+H)' _
sopropoxyphenyl)propanoic acid 586.95.
3S)-3-(3-isopropoxyphenyl)-3-( {2-[2-oxo-4 95 alculated (M-H)- = 517.27;
2-phenylethyl)pyridin- 1 (2H) ound (M-H)- = 517.2 1.
1]hexanoyl}amino)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 52 alculated (M-H)- = 487.26;
1(2H)-yl)hexanoyl]amino}-3-(3,5 ound (M-H)- = 487.20.
imethylphenyl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin, 283 alculated (M-H)- = 517.27;
1(2H)-yl)hexanoyl]amino}-3-(4-methoYy-2,5= Found (M-H) = 517.20.
imethylphenyl)propanoic acid
3S)-3-( {2-[3-(3-chlorobenzyl)-5-methyl-2-r 23 alculated (M-H)- = 551.23;
xopyridin-1(2H)-yl]hexanoyl}amino)-3-(3-11 ound (M-H)- = 551.22.
ropoxyphenyl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 250 alculated (M+H)
_
1(2H)-yl)hexanoyl]amino}-3-[3 529.23; Found (M+H)'=
trifluoromethyl)phenyl]propanoic acid 529.01.
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 27 alculated (M+H) =
1(2H)-yl)hexanoyl]amino}-3-[4 545.23;Found(M+H)`=
trifluoromethoxy)phenyl]propanoic acid 544.97.
3 S)-3-( {2-[3-(3-chlorobenzyl)-5-methyl-2 20 alculated (M+H)y =
xopyridin-1(2H)-yl]hexanoyl}amino)-3-(3 553.25; Found (M+H)+ =
isopropoxyphenyl)propanoic acid 552.95.
3S)-3-( {2-[3-(2-ethoxybenzyl)-5-methyl-2 19 alculated (M+H)'
xopyridin-1(2H)-yl]hexanoyl}amino)-3-(3 63.31; Found (M+H)" _
'sopropoxyphenyl)propanoic acid 563.17.
3S)-3-(3-isopropoxyphenyl)-3-( {2-[3-(2-4 alculated (M+H)` =
ethoxybenzyl)-5-methyl-2-oxopyridin 49.30; Found (M+H)- =
1(2H)-yl]hexanoyl } amino)propanoic acid 549.13.
CA 02361285 2001-11-07
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3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 40 alculated (M-H)- = 543.21;
1(2H)-yl)hexanoyl]amino}-3-[3 ound (M-H)- = 543.16.
trifluoromethoxy)phenyl]propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 4 alculated (M-H)- = 533.27;
1(2H)-yl)hexanoyl]amino}-3-(3-ethoxy-4 ound (M-H)- = 533.22.
ethoxyphenyl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 250 alculated (M-H)- = 549.28;
1(2H)-yl)hexanoyl]amino}-3-(4'-methyl-1,1' ound (M-H)- = 549.16.
iphenyl-4-yl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 25 2alculated (M-H)- = 551.23;
1(2H)-yl)hexanoyl]amino } -3-(4-chloro-3 ound (M-H)- = 551.20.
isopropoxyphenyl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 300 alculated (M-H)- = 565.27;
1(2H)-yl)hexanoyl]amino}-3-(4'-methoxy ound (M-H)- = 565.21.
1,1'-biphenyl-4-yl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 9 alculated (M-H)- = 503.25;
1(2H)-yl)hexanoyl]amino}-3-(4-methoxy-3 ound (M-H)- = 503.20.
ethylphenyl)propanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 10 Calculated (M-H)- = 459.23;
1(2H)-yI)hexanoyl]amino}-3-phenylpropanoic; ~ound (M-H)- = 459.18.
cid
(3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridinj 300 ~Calculated (M-H)- = 487.26;
1 (2H)-yl)hexanoyl]amino}-3-(2,5 round (M-H)- = 487.2 1.
1imethylphenyl)propanoic acid
3S)-3-( {2-[3-(2-chlorobenzyl)-5-methyl-2 15 alculated (M-H)- = 507.21;
xopyridin-1(2H)-yl]hexanoyl}amino)-3-(4~ ound (M-H)- = 507.15.
ethylphenyl)propanoic acid
3S)-3-( {2-[3-(2-chlorobenzyl)-5-methyl-2 5 alculated (M-H)- = 551.23;
xopyridin-1(2H)-yl]hexanoyl}amino)-3-(3 ound (M-H)- = 551.20.
sopropoxyphenyl)propanoic acid
3S)-3-( {2-[3-(2-chlorobenzyl)-5-methyl-2 12 alculated (M-H)- = 493.19;
xopyridin-1(2H)-yl]hexanoyl } amino)-3 ound (M-H)- = 493.12.
henylpropanoic acid
3S)-3-{[2-(3-benzyl-5-methyl-2-oxopyridin 25 alculated (M-H)- = 495.21;
1(2H)-yl)hexanoyl]amino}-3-(3,5 ound (M-H)- = 495.14.
ifluorophenyl)propanoic acid
3S)-3-(2,3-dihydro-IH-inden-5-yl)-3-({2- 63 alculated (M-H)- = 499.26;
5-methyl-2-oxo-3-(phenylmethyl)-1(2H)- ound (M-H)- = 499.23.
yridinyl]hexanoyl}amino)propanoic acid
CA 02361285 2001-11-07
83
3S)-3-( {2-[3-[(2,3-dichlorophenyl)methyl]- 68 alculated (M+H)+ _
-methyl-2-oxo-1(2H)- 586.20; Found (M+H)+ _
yridinyl]hexanoyl} amino)-3- {3-[(1- 586.88.
eth leth l)ox hen 1 ro anoic acid
3S)-3-[1,1'-biphenyl]-3-y1-3-({2-[5-methyl- 250 alculated (M-H)- = 535.26;
-oxo-3-(phenylmethyl)-1(2H)- ound (M-H)- = 535.22.
yridinyl]hexanoyl}amino)propanoic acid
3S)-3-({2-[3-{[2,4-dichloro-6- 275 alculated(M+H)+=
methyloxy)phenyl]methyl}-5-methyl-2- 16.21; Found (M+H)+=
xo-1(2H)-pyridinyl]hexanoyl } amino)-3 -{ 3- 516.88.
(1-methylethyl)oxy]phenyl } propanoic acid
3S)-3-(4-ethylphenyl)-3-( {2-[5-methyl-2- 40 alculated (M-H)- = 487.26;
xo-3-(phenylmethyl)-1(2H)- ound (M-H)- = 487.24.
yridinyl]hexanoyl}amino)propanoic acid
ti 15 3S)-3-[4-(1-methylethyl)phenyl]-3-( {2-[5- 160 alculated (M-H)- =
501.28;
ethyl-2-oxo-3-(phenylmethyl)-1(2H)- ound (M-H)- = 501.27.
yridinyl]hexanoyl}amino)propanoic acid
3S)-3-(1-methyl-iH-indol-6-yl)-3-({2-[5- 10 alculated (M-H)- = 512.25;
ethyl-2-oxo-3-(phenylmethyl)-1(2H)- found (M-H)- = 512.24.
yridinyl]hexanoyl}amino)propanoic acid
3S)-3-( {2 [5-methyl 3 (2 400 ~Calculated (M-H)- = 523.26;,=
aphthalenylmethyl)-2-oxo-1(2H)- Found (M-H)- = 523.23.
yridinyl]hexanoyl }amino)-3 -(4-eth 1 hen l) ro anoic acid
3S)-3-{3-[(1-methylethyl)oxy]phenyl}-3- 20 alculated (M-H)- = 567.29;
{2-[5-methyl-3-(2-naphthalenylmethyl)-2- Found (M-H)- = 567.26.
xo-1(2H)-
~. 'din 1 hexano 1 amino) ro anoic acid
3S)-3-(2,3-dihydro-1 H-inden-5-yl)-3-( {2- 283 alculated (M-H)- = 549.28;
5-methyl-3-(2-naphthalenylmethyl)-2-oxo- ound (M-H)- = 549.21.
1(2H)-pyridinyl]hexanoyl} amino)propanoic
icid
3S)-3-( {2-[5-methyl-3-(2- 30 alculated (M-H)- = 509.24;
aphthalenylmethyl)-2-oxo-1(2H)- ound (M-H)- = 509.19.
yridinyl]hexanoyl}amino)-3-
hen 1 ro anoic acid
2S)-2-({2-[5-methyl-2-oxo-3- >10000 alculated (M-H)- = 459.23;
phenylmethyl)-1(2H)- ound (M-H)- = 459.22.
yridinyl]hexanoyl } amino)-3-
hen 1 ro anoic acid
3S)-3-({2-[5-methyl-2-oxo-3- 10 alculated (M-H)- = 515.29;
phenylmethyl)-1(2H)- ound (M-H) = 515.28.
yridinyl]hexanoyl } amino)-3-[3-(2-
eth 1 ro 1 hen 1 ro anoic acid
CA 02361285 2001-11-07
84
3S)-3-[3-(diethylamino)phenyl]-3-( {2-[5- 10 alculated (M-H)- = 530.30;
ethyl-2-oxo-3-(phenylmethyl)-1(2H) ound (M-H)- = 530.25.
yridinyl]hexanoyl}amino)propanoic acid
3 S)-3-[3-(difluoromethyl)phenyl]-3-( {2-[5- 20 alculated (M-H)- = 509.23;
ethyl-2-oxo-3-(phenylmethyl)-1(2H)- ound (M-H)- = 509.19.
yridinyl]hexanoyl}amino)propanoic acid
3S)-3-{3-[(1-methylethyl)oxy]phenyl}-3- 5 alculated (M+H)+
{2-[5-methyl-3-[(2-methylphenyl)methyl]- 533.30; Found (M+H)' _
-oxo-1(2H)- 532.94.
yridinyl]hexanoyl } amino)propanoic acid
2R)-2-(t2-[5-methyl-2-oxo-3- 5000 alculated (M-H)- = 459.23;
phenylmethyl)-1(2H)- ound (M-H)- = 459.22.
yridinyl]hexanoyl } amino)-3-
hen 1 ro anoic acid
~ 15 3S)-3-(3-fluorophenyl)-3-({2-[5-methyl-2- 13 alculated (M-H)- = 477.21;
xo-3-(phenylmethyl)-1(2H)- ound (M-H)- = 477.20.
yridinyl]hexanoyl}amino)propanoic acid
2R)-2-( {2-[5-methyl-2-oxo-3- > 10000 alculated (M-H)- = 445.21;
phenylmethyl)-1(2H)- ound (M-H)- = 444.94.
yridinyl]hexanoyl}amino) 2-
phenylethanoic acid
3S)-3-({2-[3-[(2-chloro-4- 17 Calculated (M-H)- = 569.22;
uorophenyl)methyl]-5-methyl-2-oxo- Found (M-H)- = 569.18.
1 (2H)-pyridinyl]hexanoyl } amino)-3- {3-[(1-
eth leth l)ox hen l} ro anoic acid
3S)-3-( {2-[5-methyl-3-[(2- 25 Calculated (M+H)` _
ethylphenyl)methyl]-2-oxo-1(2H)- 4
75.26; Found (M+H)+ _
yridinyl]hexanoyl}amino)-3- 74.94.
~ _. henylpropanoic acid
3S)-3-{3-[(1-methylethyl)oxy]phenyl}-3- 10 alculated (M+H)1=
{2-[5-methyl-2-oxo-1-(phenylmethyl)-1,2 539.25; Found (M+H)+ =
ihydro-3-pyridinyl]-2- 538.91
henylacetyl}amino)propanoic acid
3S)-3-( {2-[5-methyl-3-(1- alculated (M-H)- _
aphthalenylmethyl)-2-oxo-1(2H)- 350 509.24; Found (M-H)-
yridinyl]hexanoyl } amino)-3- 509.22.
henylpropanoic acid
3S)-3-{3-[(1-methylethyl)oxy]phenyl}-3- alculated (M-H)-
{2-[5-methyl-3-(1-naphthalenylmethyl)-2- 567.29; Found (M-H)- _
xo-1(2H)- 67 567.28.
yridinyl]hexanoyl}amino)propanoic acid
CA 02361285 2001-11-07
3S)-3-{4-methyl-3-[(1- alculated (M-H)- _
ethylethyl)oxy]phenyl}-3-({2-[5-methyl-2- 15 31.29; Found (M-H)- _
xo-3-(phenylmethyl)-1(2H)- 31.26.
yridinyl)hexanoyl } amino)propanoic acid
5
CA 02361285 2001-11-07
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SEQUENCE LISTING
(1) GENERAL INFORMATION:
(i) APPLICANT: Biediger, Ronald J.; Dupre, Brian; Hamaker, Linda K.;
Holland, George W.; Kassir, Jamal M.; Li, Wen; Market, Robert V.; Nguyen,
Noel; Scott, Ian L.; Wu, Chengde; and Decker, E. Radford.
(ii) TITLE OF INVENTION: Propanoic Acid Derivatives that Inhibit
the Binding of Integrins to their Receptors
(iii) NUMBER OF SEQUENCES: 1
(iv) CORRESPONDENCE ADDRESS:
(A) ADDRESSEE: Rockey, Milnamow & Katz, Ltd.
(B) STREET: 180 N. Stetson Avenue, 2 Prudential Plaza,
Suite 4700
(C) CITY: Chicago
(D) STATE: IL
(E) COUNTRY: U.S.A.
(F) ZIP: 60601
(v) COMPUTER READABLE FORM:
(A) MEDIUM TYPE: Floppy disk
(B) COMPUTER: IBM PC compatible
(C) OPERATING SYSTEM: PC-DOS/MS-DOS
(D) SOFTWARE: Patent In Release #1.0, Version #1.30
(vi) CURRENT APPLICATION DATA:
(A) APPLICATION NUMBER:
(B) FILING DATE:
(C) CLASSIFICATION:
(viii) ATTORNEY/AGENT INFORMATION:
(A) NAME: Katz, Martin L.
(B) REGISTRATION NUMBER: 25,011
(C) REFERENCE/DOCKET NUMBER: TEX4542P0412US
(ix) TELECOMMUNICATION INFORMATION:
(A) TELEPHONE: 312-616-5400
(B) TELEFAX: 312-616-5460
(2) INFORMATION FOR SEQ ID NO:1:
CA 02361285 2001-11-07
87
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 26 amino acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: protein
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1:
Cys Asp Glu Leu Pro Gin Leu Val Thr Leu Pro His Pro Asn Leu His
1 5 10 15
Gly Pro Glu Ile Leu Asp Val Pro Ser Thr
25
~: .
CA 02361285 2008-10-23
88
The present invention is illustrated by way of the foregoing description
and examples. The foregoing description is intended as a non-limiting
illustration, since many variations will become apparent to those skilled in
the
art in view thereof. It is intended that all such variations within the scope
and
spirit of the appended claims be embraced thereby.
Changes can be made in the composition, operation and arrangement of
the method of the present invention described herein without departing from
the
concept and scope of the invention as defined in the following claims:
