U.S. patent application number 11/594432 was filed with the patent office on 2007-03-08 for vla-4 inhibitor compounds.
This patent application is currently assigned to Daiichi Pharmaceutical Co., Ltd.. Invention is credited to John J. Baldwin, Jun Chiba, Shin Iimura, Nobuo Machinaga, Edward McDonald, Kevin Joseph Moriarty, Atsushi Nakayama, Christopher Ronald Sarko, Yoshiyuki Yoneda.
Application Number | 20070054909 11/594432 |
Document ID | / |
Family ID | 27488233 |
Filed Date | 2007-03-08 |
United States Patent
Application |
20070054909 |
Kind Code |
A1 |
Baldwin; John J. ; et
al. |
March 8, 2007 |
VLA-4 inhibitor compounds
Abstract
Compounds that selectively inhibit the binding of ligands to
.alpha.4.beta.1 integrin (VLA-4) and methods for their preparation
are disclosed. In one embodiment, compounds of the invention are
represented by Formula I: ##STR1## As selective inhibitors of VLA-4
mediated cell adhesion, compounds of the present invention are
useful in the treatment of conditions associated with such
adhesion, including, but not limited to, such conditions as
inflammatory and autoimmune responses, diabetes, asthma, psoriasis,
inflammatory bowel disease, transplantation rejection, and tumor
metastasis. Also disclosed are methods of inhibiting VLA-4 mediated
cell adhesion and methods of treating conditions associated with
LA-4 mediated cell adhesion.
Inventors: |
Baldwin; John J.; (Gwynedd,
PA) ; McDonald; Edward; (Surrey, GB) ;
Moriarty; Kevin Joseph; (Norristown, PA) ; Sarko;
Christopher Ronald; (New Milford, CT) ; Machinaga;
Nobuo; (Tokyo, JP) ; Nakayama; Atsushi;
(Tokyo, JP) ; Chiba; Jun; (Tokyo, JP) ;
Iimura; Shin; (Tokyo, JP) ; Yoneda; Yoshiyuki;
(Tokyo, JP) |
Correspondence
Address: |
HESLIN ROTHENBERG FARLEY & MESITI PC
5 COLUMBIA CIRCLE
ALBANY
NY
12203
US
|
Assignee: |
Daiichi Pharmaceutical Co.,
Ltd.
Tokyo
NJ
Pharmacopeia Drug Discovery, Inc.
Cranbury
|
Family ID: |
27488233 |
Appl. No.: |
11/594432 |
Filed: |
November 8, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10787905 |
Feb 26, 2004 |
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11594432 |
Nov 8, 2006 |
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10034585 |
Dec 28, 2001 |
6756378 |
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10787905 |
Feb 26, 2004 |
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PCT/US00/18079 |
Jun 30, 2000 |
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10034585 |
Dec 28, 2001 |
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60141601 |
Jun 30, 1999 |
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60141602 |
Jun 30, 1999 |
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60141692 |
Jun 30, 1999 |
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Current U.S.
Class: |
514/237.8 ;
514/254.01; 514/326; 514/343; 514/365; 514/381; 514/415; 514/423;
544/372; 546/207; 546/279.1; 548/482; 548/530 |
Current CPC
Class: |
C07D 295/185 20130101;
C07D 207/22 20130101; C07C 275/42 20130101; C07D 209/12 20130101;
C07D 295/13 20130101; C07D 401/06 20130101; C07D 307/33 20130101;
C07D 277/28 20130101; C07D 211/34 20130101; C07D 401/14 20130101;
C07D 213/55 20130101; C07D 207/12 20130101; C07D 491/04 20130101;
C07D 211/38 20130101; C07D 207/14 20130101; C07D 311/20 20130101;
C07C 2601/02 20170501; C07D 401/12 20130101; C07D 207/24 20130101;
C07D 207/16 20130101; C07D 207/08 20130101; C07D 413/06 20130101;
C07D 403/02 20130101; C07D 277/48 20130101; C07D 217/02 20130101;
C07D 277/04 20130101; C07D 211/26 20130101; C07D 207/09
20130101 |
Class at
Publication: |
514/237.8 ;
514/381; 514/423; 514/415; 514/254.01; 514/326; 514/343; 514/365;
544/372; 546/207; 546/279.1; 548/482; 548/530 |
International
Class: |
A61K 31/5375 20070101
A61K031/5375; A61K 31/496 20070101 A61K031/496; A61K 31/4439
20070101 A61K031/4439; A61K 31/426 20070101 A61K031/426; A61K
31/401 20070101 A61K031/401; A61K 31/405 20070101 A61K031/405; C07D
403/02 20070101 C07D403/02 |
Claims
1. A compound represented by Formula I, or a salt thereof,
##STR1957## wherein W is chosen from aryl group, substituted aryl
group, heteroaryl group and substituted heteroaryl group; W.sup.1
is chosen from arylene group, substituted arylene group,
heteroarylene group and substituted heteroarylene group; A is
chosen from .dbd.O, .dbd.S and .dbd.NH; R is chosen from a direct
bond, alkyenylene group and --(CH.sub.2).sub.n--, wherein n is
chosen from 1 and 2; X is chosen from --C(O)--, --CH.sub.2-- and
S(O).sub.2; M is chosen from ##STR1958## wherein ##STR1959## is a
divalent 4-, 5-, 6- or 7-membered heterocyclic moiety, other than
pyrrolidine or thiazolidine, wherein the nitrogen atom is the point
of attachment to X; R.sup.1, R.sup.2 and R.sup.3 are independently
chosen from --H, --OH, --NH.sub.2, halogen atom, alkyl group,
substituted alkyl group, aryl group, substituted aryl group, alkoxy
group, substituted alkoxy group, monoalkylamino group, substituted
monoalkylamino group, dialkylamino group, substituted dialkylamino
group, cycloalkylamino group, substituted cycloalkylamino group,
alkylsulfonylamino group, substituted alkylsulfonylamino group,
arylsulfonylamino group, substituted arylsulfonylamino group,
aryloxy group, substituted aryloxy group, heteroaryloxy group,
substituted heteroaryloxy group, benzyloxy group and substituted
benzyloxy group, or two of R.sup.1, R.sup.2 and R.sup.3 taken
together may form a 3-, 4-, 5-, 6-, or 7-membered carbocyclic or
heterocyclic residues optionally substituted with from 1 to 3
substituents chosen independently from --OH, halogen atom,
--NH.sub.2, alkyl group, alkoxy group, aryl group, aryloxy group,
alkylamino group, benzyloxy group and heteroaryl group; R.sup.4 is
chosen from --H and lower alkyl group; Y is a direct bond or a
divalent radical chosen from --C(O)--, --C(O)NH--, alkenylene
group, alkynylene group and --(CH.sub.2).sub.kY.sup.2, wherein k is
chosen from 1, 2 and 3; and Y.sup.2 is a direct bond or a divalent
radical chosen from --O--, --S--, --S(O), --S(O).sub.2-- and
--NY.sup.3--, wherein Y.sup.3 is chosen from --H and lower alkyl
group; Z is chosen from arylene group, substituted arylene group,
heterocyclylene group, substituted heterocyclylene group,
cycloalkylene group and substituted cycloalkylene group; A.sup.1 is
a direct bond or a divalent radical chosen from alkenylene group,
alkynylene group, --(CH.sub.2).sub.t-- and --O(CH.sub.2), wherein t
is chosen from 1, 2 and 3; and v is chosen from 0, 1, 2, and 3; and
R.sup.5 is chosen from --OH, lower alkoxy group, --N(H)OH,
##STR1960## wherein ##STR1961## is a divalent 4-, 5-, 6- or
7-membered heterocyclic moiety, other than pyrrolidine or
thiazolidine, wherein the nitrogen atom is the point of attachment
to X; R.sup.6 and R.sup.7 are independently chosen from --H, --OH,
halogen atom, alkyl group and alkoxy group; Y.sup.1 is a divalent
radical chosen from --O--, --S--, --S(O)--, --S(O).sub.2-- and
--NY.sup.4--, wherein Y.sup.4 is chosen from --H and lower alkyl
group; Z.sup.1 is a divalent radical chosen from arylene group,
substituted arylene group, heterocyclylene group, substituted
heterocyclylene group, cycloalkylene group and substituted
cycloalkylene group; A.sup.2 is a direct bond or a divalent radical
chosen from alkenylene group, alkynylene group and
--(CH.sub.2).sub.e wherein e is chosen from 1, 2 and 3; and R.sup.8
is chosen from --OH, lower alkoxy group, --N(H)OH, ##STR1962##
wherein L is ##STR1963## wherein ##STR1964## is a divalent 4-, 5-,
6- or 7-membered heterocyclic moiety, other than pyrrolidine or
thiazolidine, optionally substituted with from 1 to 3 substitutents
chosen independently from alkyl group, alkoxy group, hydroxyalkyl
group, --OH, benzyloxy group, --NH.sub.2, halogen atom, aryl group
and heteroaryl group, said moiety may be fused to 1 or 2 additional
carbocyclic or heterocyclic residues optionally substituted with
from 1 to 3 substitutents chosen independently from alkyl group,
aryloxy group, alkoxy group, hydroxyalkyl group, --OH, benzyloxy
group, --NH.sub.2, halogen atom, aryl group and heteroaryl group; m
and q are independently chosen from 0, 1, 2 and 3; X.sup.1 is
chosen from --CH.dbd. and --N.dbd.; R.sup.9 is chosen from --H and
lower alkyl group; R.sup.10 is chosen from --COOH, lower
alkoxycarbonyl group, ##STR1965## and Z.sup.2 is chosen from --H,
COOH and lower alkoxycarbonyl group; and
-R.sup.11-Z.sup.3-Q.sup.2-L.sup.1, wherein R.sup.11 is chosen from
--O--, ##STR1966## and --NR.sup.12-- wherein R.sup.12 is chosen
from --H, alkyl group, substituted alkyl group, cycloalkyl group,
substituted cycloalkyl group, aryl group, substituted aryl group,
benzyl group, substituted benzyl group, lower alkenyl group,
substituted lower alkenyl group and lower alkynyl group and the
left hand bond is the point of attachment to --X-- and the right
hand bond is the point of attachment to -Z.sup.3; Z.sup.3 is chosen
from a direct bond, a divalent aliphatic hydrocarbon moiety having
1 to 12 carbon atoms, wherein one or more carbon atoms may be
replaced with --O-- or --NR.sup.13--, wherein R.sup.13 is chosen
from --H and lower alkyl group, and one or more hydrogen atoms
attached to an aliphatic carbon atom may be replaced with lower
alkyl group; and ##STR1967## wherein x is chosen from 0 and 1; y is
chosen from 1, 2, and 3; and R.sup.14 is chosen from --H, --OH and
halogen atom, ##STR1968## when R.sup.11 is --NR.sup.2, -Z.sup.4-
wherein Z.sup.4 is chosen from ##STR1969## wherein R.sup.14a is
chosen from --H, --OH, lower alkyl group and halogen atom;
##STR1970## 'wherein the left hand bond is the point of attachment
to R.sup.11 and the right hand bond is the point of attachment to
Q.sup.2; Q.sup.2 is a divalent radical chosen from arylene group,
substituted arylene group, heterocyclylene group, substituted
heterocyclylene group, cycloalkylene group, substituted
cycloalkylene group, ##STR1971## wherein R.sup.15 and R.sup.16 are
independently chosen from --H, halogen atom and lower alkyl group;
and ##STR1972## wherein R.sup.17 and R.sup.18 are independently
chosen from --H, lower alkyl group, substituted lower alkyl group
and lower alkenyl group; and L.sup.1 is chosen from --COOH and
--COOR.sup.19 wherein R.sup.19 is a lower alkyl group.
2. A compound according to claim 1, or a salt thereof, wherein M is
##STR1973##
3. A compound according to claim 1, or a salt thereof, wherein M is
##STR1974##
4. A compound according to claim 1, or a salt thereof, wherein M is
##STR1975##
5. A compound according to claim 1, or a salt thereof, wherein M is
-R.sup.11-Z.sup.3-Q.sup.2-L.sup.1.
6. A compound according to claim 2, or a salt thereof, wherein at
least one radical of R.sup.1, R.sup.2 and R.sup.3 is --OH or
halogen atom.
7. A compound according to claim 6, or a salt thereof, wherein A is
.dbd.O, R is --(CH.sub.2).sub.n-- and X is --C(O)--.
8. A compound according to claim 7, or a salt thereof, wherein Y is
chosen from alkenylene group, alkynylene group and
--(CH.sub.2).sub.kY.sup.2; Y.sup.2 is chosen from a direct bond,
--O--, --S(O) and --NY.sup.3--; and Y.sup.3 is --H.
9. A compound according to claim 8, or a salt thereof, wherein Y is
chosen from --O-- and --NY.sup.3--.
10. A compound according to claim 2, or a salt thereof, wherein W
is unsubstituted phenyl group or phenyl group having one or two
substituents chosen from lower alkyl group and halogen atom at the
ortho positions thereof.
11. A compound according to claim 10, or a salt thereof, wherein
W.sup.1 is unsubstituted phenylene group or phenylene group having
a substituent chosen from methoxy group, lower alkyl group and
halogen atom at the ortho position to the --NH-- thereof.
12. A compound according to claim 2, or a salt thereof, wherein
W.sup.1 is phenylene group having a substituent chosen from methoxy
group, lower alkyl group and halogen atom at the ortho position to
the --NH-- thereof and having 1 to 3 substituents chosen from lower
alkyl group and halogen atom.
13. A compound according to claim 2, or a salt thereof, wherein
A.sup.1 is a direct bond or --(CH.sub.2).sub.t--.
14. A compound according to claim 13, or a salt thereof, wherein
A.sup.1 is a direct bond.
15. A compound according to claim 14, or a salt thereof, wherein
R.sup.5 is --OH.
16. A compound according to claim 2, or a salt thereof, wherein W
is unsubstituted phenyl group or phenyl group having one or two
substituents chosen from lower alkyl group and halogen atom at the
ortho positions thereof; W.sup.1 is unsubstantiated phenylene group
or phenylene group having a substituent chosen from methoxy group,
lower alkyl group and halogen atom at the ortho position to the
--NH-- thereof; R is --CH.sub.2--; X is --C(O)-- and R.sup.5 is
--OH.
17. (canceled)
18. (canceled)
19. A compound according to claim 2, or a salt thereof, wherein
R.sup.5 is lower alkoxy group.
20. A method of inhibiting cell adhesion in a mammal, said method
comprising administering to said mammal an effective amount of a
compound according to claim 1.
21. A method of treating a condition associated with VLA-4 mediated
cell adhesion in a mammal, aid method comprising administering to
said mammal an effective amount of a compound according to claim
1.
22. A method according to claim 21, wherein the condition
associated with VLA-4 mediated cell adhesion is chosen from
inflammatory responses, autoimmune responses and tumor
metastasis.
23. A method according to claim 21, wherein the condition
associated with VLA-4 mediated cell adhesion is chosen from asthma,
diabetes, arthritis, psoriasis, multiple sclerosis, inflammatory
bowel disease and transplantation rejection.
24. (canceled)
25. (canceled)
26. A pharmaceutical composition comprising as a therapeutic agent,
a compound according to claim 1 and a pharmaceutically acceptable
carrier or excipient.
27. A pharmaceutical composition according to claim 26, further
comprising one or more additional therapeutic agents.
28. A pharmaceutical composition according to claim 27, wherein
said one or more additional therapeutic agents are chosen from the
group consisting of antiinflammatory, antirheumatic,
corticosteroid, immunosuppressive, antipsoriatic, bronchodilator,
antiasthmatic and antidiabetic agents.
29. A pharmaceutical composition according to claim 28, wherein one
of said one or more additional therapeutic agents is an
antiinflammatory agent.
30. A pharmaceutical composition according to claim 29, wherein
said antiinflammatory agent is chosen from a steroid and an
NSAID.
31. A compound according to claim 16, or a salt thereof, wherein
the compound is: ##STR1976##
Description
FIELD OF THE INVENTION
[0001] The present invention relates to compounds that selectively
inhibit the binding of ligands to the adhesion receptor,
.alpha..sub.4.beta..sub.1 integrin, also known as VLA-4. Compounds
of the present invention are useful in the treatment and prevention
of pathologies associated with VLA-4 mediated cell adhesion, such
as inflammatory and autoimmune diseases, and tumor metastasis.
BACKGROUND OF THE INVENTION
[0002] A primary feature of such pathologies as inflammation and
autoimmune diseases is the accumulation of activated leukocytes in
affected tissues. The process by which leukocytes transmigrate from
the circulation at a site of inflammation involves a cascade of
interactions that can be divided into four major steps: tethering
and rolling, activation, firm adhesion, and transmigration
(Springer, T., Ann. Rev. Physiol. 57:827 (1995)). Initially,
leukocytes are lightly tethered to the endothelium and roll along
its surface. This is followed by cell activation, mediated by
soluble chemotactic stimuli, which initiates the development of a
firmer bond between individual leukocytes and endothelial cells.
The firm bond then results in the successful adhesion and
transmigration of the leukocytes through endothelial cell
junctions. The steps occur in series and each is essential for
transmigration to occur. This also means that transmigration can be
modulated at each step, thus providing a number of potential
targets for pharmacological inhibition.
[0003] The receptors involved in leukocyte migration have, to a
large extent, been characterized as belonging to particular cell
adhesion molecule families (Carlos and Harlan, Blood, 84:2068
(1994)). The initial attachment and rolling step is mediated by a
family of adhesion receptors referred to as selectins. Firm
adhesion is mediated by interaction of leukocyte surface integrins
with molecules of the immunoglobulin superfamily expressed on the
surface of the endothelium. Both integrins and the
immunoglobulin-type adhesion molecules are also primarily involved
in leukocyte transmigration. After transmigration, the leukocytes
rely on integrins to traverse through the extracellular matrix and
remain at the site of inflammation.
[0004] Integrins are a large family of heterodimeric glycoproteins
composed of two noncovalently associated subunits, .alpha. and
.beta. (Hynes, R., Cell, 69:11 (1992)). There are at least 16
different .alpha. subunits (.alpha..sub.1-.alpha..sub.9,
.alpha..sub.L, .alpha..sub.M, .alpha..sub.D, .alpha..sub.X,
.alpha..sub.E, .alpha..sub.IIb, .alpha..sub.V) and at least 9
different .beta. (.beta..sub.1-.beta..sub.9) subunits. Integrins
are divided into sub-families, based upon the .beta. subunit.
Leukocytes express a number of different integrins, including
.alpha..sub.4.beta..sub.1, .alpha..sub.5.beta..sub.1,
.alpha..sub.6.beta..sub.1, .alpha..sub.4.beta..sub.7,
.alpha..sub.L.beta..sub.2, .alpha..sub.X.beta..sub.2, and
.alpha..sub.V.beta..sub.3.
[0005] .alpha..sub.4.beta..sub.1 integrin, also known as very late
antigen-4 (VLA-4) or CD49d/CD29, is expressed on monocytes,
lymphocytes, eosinophils, and basophils, all of which are key
effector cells in various inflammatory disorders (Helmer, M., Ann.
Rev. Immunol., 8:365 (1990)). .alpha..sub.4.beta..sub.1 integrin
serves as a receptor for vascular cell adhesion molecule-1
(VCAM-1), as well as to the extracellular protein fibronectin (FN)
(Elices et al., Cell, 60:577 (1990)). Anti-inflammatory effects and
delayed disease progression have been demonstrated after in vivo
monoclonal antibody blockade of the
.alpha..sub.4.beta..sub.1/VCAM-1 pathway (Lobb et al., J. Clin.
Invest., 94:1722-28 (1994)). In a guinea pig model of pulmonary
inflammation, anti-.alpha..sub.4 inhibited both antigen-induced
bronchial hyperreactivity and leukocyte recruitment in
bronchoalveolar lavage fluid (Pretolani et al., J. Exp. Med.,
180:795 (1994)). Antibodies to .alpha..sub.4 or VCAM-1, prevented
antigen-induced eosinophil infiltration of the mouse trachea
(Nakajima et al., J. Exp. Med. 179:1145 (1994)). .alpha..sub.4 or
VCAM-1 monoclonal antibody treatment also delayed or prevented
cutaneous delayed hypersensitivity response in mice and monkeys
(Chisholm et al., Eur. J. Immunol., 23:682 (1993); Silber et al, J.
Clin. Invest. 93:1554 (1993); cardiac allograft rejection in mice,
accompanied by specific immunosuppression (Isobe et al., J.
Immunol., 153:5810 (1994); graft-versus-host disease in mice after
bone marrow transfer (Yang et al., Proc. Natl. Acad. Sci. USA,
90:10494, (1993); and experimental autoimmune encephalomyelitis in
rats and mice (Yednock et al., Nature, 356:63 (1992); Baron et al.,
J. Exp. Med. 177:57 (1993)).
[0006] Rational drug design studies have produced soluble VCAM-Ig
fusion protein containing the two N-terminal domains of human
VCAM-1 fused to a human IgG1 constant region. In vivo
administration of the fusion protein significantly delays the onset
of adoptively transferred autoimmune diabetes in nonobese diabetic
mice (Jakubowski et al., J. Immunol. 155:938 (1995)). Another
approach has used three-dimensional crystallographic structures of
VCAM-1 fragments to synthesize cyclic peptide antagonists that
closely mimicked the .alpha..sub.4 integrin binding loop in domain
1 of VCAM-1. Synthetic VCAM-1 peptide CQIDSPC, was able to inhibit
the adhesion of VLA-4-expressing cells to purified VCAM-1 (Wang et
al., Proc. Natl. Acad. Sci. USA, 92:5714 (1995)).
[0007] An additional strategy is to block the binding of
.alpha..sub.4.beta..sub.1 to its other counter receptor, that is,
an alternatively spliced region of fibronectin containing the
connecting segment-1 (CS-1) motif (E. A. Wayner, J. Cell. Biol.,
116:489 (1992)). A synthetic CS-1 tetrapeptide (phenylacetic
acid-Leu-Asp-Phe-d-Pro-amide) inhibited VLA-4-mediated lymphocyte
adherence in vitro and reduced accelerated coronary arteriopathy in
rabbit cardiac allografts (Molossi et al., J. Clin. Invest.,
95:2601 (1995)). Each of these studies provide evidence that
selective inhibition of .alpha..sub.4.beta..sub.1/VCAM-1 mediated
adhesion is a proven strategy in the treatment of autoimmune and
allergic inflammatory diseases.
[0008] Moreover, while U.S. Pat. No. 5,821,231 and PCT Applications
WO 96/22966, WO 97/03094, WO 98/04247 and WO 98/04913 describe
compounds exhibiting VLA-4 inhibitory activity in in vitro binding
assays, none of the described compounds have exhibited efficacy in
oral administration.
[0009] Accordingly, despite these advances, there remains a need
for small, non-peptidic, specific inhibitors of VLA-4 dependent
cell adhesion that are orally bioavailable and that are suitable
for the long-term treatment of chronic inflammatory diseases and
other pathologies associated with leukocyte migration and
adhesion.
SUMMARY OF THE INVENTION
[0010] The compounds of the present invention selectively inhibit
the binding of ligands to .alpha..sub.4.beta..sub.1 and therefore,
are useful for inhibition, prevention and suppression of
VLA-4-mediated cell adhesion and the pathologies associated with
that adhesion, such as, for example, inflammation, asthma,
arthritis, diabetes, autoimmune responses, multiple sclerosis,
psoriasis, transplantation rejection, and tumor metastasis.
[0011] In one embodiment, the present invention provides a compound
represented by Formula I, or a salt thereof, ##STR2##
[0012] wherein [0013] W is chosen from aryl group, substituted aryl
group, heteroaryl group and substituted heteroaryl group; [0014]
W.sup.1 is chosen from arylene group, substituted arylene group,
heteroarylene group and substituted heteroarylene group; [0015] A
is chosen from .dbd.O, .dbd.S and .dbd.NH; [0016] R is chosen from
a direct bond, alkyenylene group and --(CH.sub.2).sub.n--, [0017]
wherein [0018] n is chosen from 1 and 2; [0019] X is chosen from
--C(O)--, --CH.sub.2-- and S(O).sub.2; [0020] M is chosen from
##STR3## [0021] wherein ##STR4## [0022] is a divalent 4-, 5-, 6- or
7-membered heterocyclic moiety, wherein the nitrogen atom is the
point of attachment to X; [0023] R.sup.1, R.sup.2 and R.sup.3 are
independently chosen from --H, --OH, --NH.sub.2, halogen atom,
alkyl group, substituted alkyl group, aryl group, substituted aryl
group, alkoxy group, substituted alkoxy group, monoalkylamino
group, substituted monoalkylamino group, dialkylamino group,
substituted dialkylamino group, cycloalkylamino group, substituted
cycloalkylamino group, alkylsulfonylamino group, substituted
alkylsulfonylamino group, arylsulfonylamino group, substituted
arylsulfonylamino group, aryloxy group, substituted aryloxy group,
heteroaryloxy group, substituted heteroaryloxy group, benzyloxy
group and substituted benzyloxy group, or [0024] two of R.sup.1,
R.sup.2 and R.sup.3 taken together may form a 3-, 4-, 5-, 6-, or
7-membered carbocyclic or heterocyclic residues optionally
substituted with from 1 to 3 substituents chosen independently from
--OH, halogen atom, --NH.sub.2, alkyl group, alkoxy group, aryl
group, aryloxy group, alkylamino group, benzyloxy group and
heteroaryl group; [0025] R.sup.4 is chosen from --H and lower alkyl
group; [0026] Y is a direct bond or a divalent radical chosen from
--C(O)--, --C(O)NH--, alkenylene group, alkynylene group and
--(CH.sub.2).sub.kY.sup.2, [0027] wherein [0028] k is chosen from
1, 2 and 3; and [0029] Y.sup.2 is a direct bond or a divalent
radical chosen from --O--, --S--, --S(O), --S(O).sub.2-- and
--NY.sup.3--, [0030] wherein [0031] Y.sup.3 is chosen from --H and
lower alkyl group; [0032] Z is chosen from arylene group,
substituted arylene group, heterocyclylene group, substituted
heterocyclylene group, cycloalkylene group and substituted
cycloalkylene group; [0033] A.sup.1 is a direct bond or a divalent
radical chosen from alkenylene group, alkynylene group,
--(CH.sub.2).sub.t-- and --O(CH.sub.2).sub.v, [0034] wherein [0035]
t is chosen from 1, 2 and 3; and [0036] v is chosen from 0, 1, 2,
and 3; and [0037] R.sup.5 is chosen from --OH, lower alkoxy group,
--N(H)OH, ##STR5## [0038] wherein ##STR6## [0039] is a divalent 4-,
5-, 6- or 7-membered heterocyclic moiety, wherein the nitrogen atom
is the point of attachment to X; [0040] R.sup.6 and R.sup.7 are
independently chosen from --H, --OH, halogen atom, alkyl group and
alkoxy group; [0041] Y.sup.1 is a divalent radical chosen from
--O--, --S--, --S(O)--, --S(O).sub.2-- and --NY.sup.4--, [0042]
wherein [0043] Y.sup.4 is chosen from --H and lower alkyl group;
[0044] Z.sup.1 is a divalent radical chosen from arylene group,
substituted arylene group, heterocyclylene group, substituted
heterocyclylene group, cycloalkylene group and substituted
cycloalkylene group; [0045] A.sup.2 is a direct bond or a divalent
radical chosen from alkenylene group, alkynylene group and
--(CH.sub.2).sub.e [0046] wherein [0047] e is chosen from 1, 2 and
3; and [0048] R.sup.8 is chosen from --OH, lower alkoxy group,
--N(H)OH, ##STR7## [0049] wherein [0050] L is ##STR8## [0051]
wherein ##STR9## [0052] is a divalent 4-, 5-, 6- or 7-membered
heterocyclic moiety, optionally substituted with from 1 to 3
substitutents chosen independently from alkyl group, alkoxy group,
hydroxyalkyl group, --OH, benzyloxy group, --NH.sub.2, halogen
atom, aryl group and heteroaryl group, said moiety may be fused to
1 or 2 additional carbocyclic or heterocyclic residues optionally
substituted with from 1 to 3 substitutents chosen independently
from alkyl group, aryloxy group, alkoxy group, hydroxyalkyl group,
--OH, benzyloxy group, --NH.sub.2, halogen atom, aryl group and
heteroaryl group; [0053] m and q are independently chosen from 0,
1, 2 and 3; [0054] X.sup.1 is chosen from --CH.dbd. and --N.dbd.;
[0055] R.sup.9 is chosen from --H and lower alkyl group; [0056]
R.sup.10 is chosen from --COOH, lower alkoxycarbonyl group,
##STR10## [0057] is chosen from --H, COOH and lower alkoxycarbonyl
group; and [0058] wherein [0059] R.sup.11 is chosen from --O--,
##STR11## [0060] and --NR.sup.12-- [0061] wherein [0062] R.sup.12
is chosen from --H, alkyl group, substituted alkyl group,
cycloalkyl group, substituted cycloalkyl group, aryl group,
substituted aryl group, benzyl group, substituted benzyl group,
lower alkenyl group, substituted lower alkenyl group and lower
alkynyl group [0063] the left hand bond is the point of attachment
to --X-- and the right hand bond is the point of attachment to
-Z.sup.3; [0064] Z.sup.3 is chosen from a direct bond, a divalent
aliphatic hydrocarbon moiety having 1 to 12 carbon atoms, [0065]
wherein [0066] one or more carbon atoms may be replaced with --O--
or --NR.sup.13-- [0067] wherein [0068] R.sup.13 is chosen from --H
and lower alkyl group, and [0069] one or more hydrogen atoms
attached to an aliphatic carbon atom may be replaced with lower
alkyl group; and ##STR12## [0070] wherein [0071] x is chosen from 0
and 1; [0072] y is chosen from 1, 2, and 3; and [0073] R.sup.14 is
chosen from --H, --OH and halogen atom, ##STR13## [0074] when
[0075] R.sup.11 is --NR.sup.12, -Z.sup.4- [0076] wherein [0077]
-Z.sup.4- [0078] wherein [0079] Z.sup.4 is chosen from ##STR14##
[0080] wherein [0081] R.sup.14a is chosen from --H, --H, lower
alkyl group and halogen atom; ##STR15## [0082] wherein the left
hand bond is the point of attachment to R.sup.11 and the right hand
bond is the point of attachment to Q.sup.2; [0083] Q.sup.2 is a
divalent radical chosen from arylene group, substituted arylene
group, heterocyclylene group, substituted heterocyclylene group,
cycloalkylene group, substituted cycloalkylene group, ##STR16##
[0084] wherein R.sup.15 and R.sup.16 are independently chosen from
--H, halogen atom and lower alkyl group; and ##STR17## [0085]
wherein R.sup.17, R.sup.18 are independently chosen from --H, lower
alkyl group, substituted lower alkyl group and lower alkenyl group;
and [0086] L.sup.1 is chosen from --COOH and --COOR.sup.19 [0087]
wherein [0088] R.sup.19 is a lower alkyl group.
[0089] In a preferred embodiment of Formula I, M is ##STR18## In
this embodiment, more preferred compounds are those wherein A is
.dbd.O, R is --(CH.sub.2).sub.n-- and X is --C(O)--. Y is
preferably chosen from alkenylene group, alkynylene group,
--(CH.sub.2).sub.kY.sup.2, --CH.sub.2S(O)-- and --CH.sub.2O--, and
more preferably, Y is --CH.sub.2O--.
[0090] Preferred compounds of this embodiment are those wherein W
is unsubstituted phenyl group or phenyl group having one or two
substituents chosen from lower alkyl group and halogen atom at the
ortho positions thereof. W.sup.1 is preferably unsubstituted
phenylene group or phenylene group having a substituent chosen from
methoxy group, lower alkyl group and halogen atom at the ortho
position to --NH--.
[0091] In preferred compounds of this embodiment, A is preferably
.dbd.O and A.sup.1 is a direct bond or --(CH.sub.2).sub.t--. More
preferred compounds are those wherein A.sup.1 is a direct bond and
R.sup.5 is --OH,
[0092] Preferred compounds of Formula I, wherein M is ##STR19##
[0093] and A is .dbd.O are represented in Table 1. With respect to
the representation of --W.sup.1, the lower bond is the point of
attachment to --NH-- and the upper bond is the point of attachment
to --R--. The entry entitled --R--R.sup.5 depicts that portion of
the particular compound represented by TABLE-US-00001 TABLE 1
##STR20## Mass Spectrum (M.sup.+ + 1) W-- --W.sup.1-- -- --R-- --
--R.sup.5 502.6 ##STR21## ##STR22## ##STR23## 516.7 ##STR24##
##STR25## ##STR26## 486.7 ##STR27## ##STR28## ##STR29## 508.7
##STR30## ##STR31## ##STR32## 507.0 ##STR33## ##STR34## ##STR35##
501.6 ##STR36## ##STR37## ##STR38## 486.6 ##STR39## ##STR40##
##STR41## 553.6 ##STR42## ##STR43## ##STR44## 486.6 ##STR45##
##STR46## ##STR47## 579.7 ##STR48## ##STR49## ##STR50## 552.7
##STR51## ##STR52## ##STR53## 496.6 ##STR54## ##STR55## ##STR56##
500.6 ##STR57## ##STR58## ##STR59## 512.6 ##STR60## ##STR61##
##STR62## 527.7 ##STR63## ##STR64## ##STR65## 484 ##STR66##
##STR67## ##STR68## 486 ##STR69## ##STR70## ##STR71## 488 ##STR72##
##STR73## ##STR74## 504 ##STR75## ##STR76## ##STR77## 518 ##STR78##
##STR79## ##STR80## 502 ##STR81## ##STR82## ##STR83## 508 ##STR84##
##STR85## ##STR86## 498 ##STR87## ##STR88## ##STR89## 512 ##STR90##
##STR91## ##STR92## 526 ##STR93## ##STR94## ##STR95## 532 ##STR96##
##STR97## ##STR98## 514 ##STR99## ##STR100## ##STR101## 500
##STR102## ##STR103## ##STR104## 516 ##STR105## ##STR106##
##STR107## 534 ##STR108## ##STR109## ##STR110## 566 ##STR111##
##STR112## ##STR113## 566 ##STR114## ##STR115## ##STR116## 534
##STR117## ##STR118## ##STR119## 550 ##STR120## ##STR121##
##STR122## 517 ##STR123## ##STR124## ##STR125## ##STR126##
##STR127## ##STR128## 548 ##STR129## ##STR130## ##STR131## 562
##STR132## ##STR133## ##STR134## 552 ##STR135## ##STR136##
##STR137## 586 ##STR138## ##STR139## ##STR140## 563 ##STR141##
##STR142## ##STR143## 533 ##STR144## ##STR145## ##STR146## 516
##STR147## ##STR148## ##STR149## 532 ##STR150## ##STR151##
##STR152## 562 ##STR153## ##STR154## ##STR155## 542 ##STR156##
##STR157## ##STR158## 561 ##STR159## ##STR160## ##STR161## 536
##STR162## ##STR163## ##STR164## 552 ##STR165## ##STR166##
##STR167## 519 ##STR168## ##STR169## ##STR170## 654 ##STR171##
##STR172## ##STR173## 577 ##STR174## ##STR175## ##STR176## 564
##STR177## ##STR178## ##STR179## 549 ##STR180## ##STR181##
##STR182## 566 ##STR183## ##STR184## ##STR185## 575 ##STR186##
##STR187## ##STR188## 553 ##STR189## ##STR190## ##STR191## 523
##STR192## ##STR193## ##STR194## 524 ##STR195## ##STR196##
##STR197## 517 ##STR198## ##STR199## ##STR200## 535 ##STR201##
##STR202## ##STR203## 521 ##STR204## ##STR205## ##STR206## 580
##STR207## ##STR208## ##STR209## 550 ##STR210## ##STR211##
##STR212## 537 ##STR213## ##STR214## ##STR215## 582 ##STR216##
##STR217## ##STR218## 553 ##STR219## ##STR220## ##STR221## 674
##STR222## ##STR223## ##STR224## 598 ##STR225## ##STR226##
##STR227## 626 ##STR228## ##STR229## ##STR230## 599 ##STR231##
##STR232## ##STR233## 554 ##STR234## ##STR235## ##STR236## 537
##STR237## ##STR238## ##STR239## 582 ##STR240## ##STR241##
##STR242## 535 ##STR243## ##STR244## ##STR245## 639 ##STR246##
##STR247## ##STR248## 646 ##STR249## ##STR250## ##STR251## 629
##STR252## ##STR253## ##STR254## 538 ##STR255## ##STR256##
##STR257## 554 ##STR258## ##STR259## ##STR260## 583 ##STR261##
##STR262## ##STR263## 536 ##STR264## ##STR265## ##STR266## 556
##STR267## ##STR268## ##STR269## 538 ##STR270## ##STR271##
##STR272## 518 ##STR273## ##STR274## ##STR275## 601 ##STR276##
##STR277## ##STR278## 516 ##STR279## ##STR280## ##STR281## 534
##STR282## ##STR283## ##STR284## 554 ##STR285## ##STR286##
##STR287## 542 ##STR288## ##STR289## ##STR290## 561 ##STR291##
##STR292## ##STR293## 536 ##STR294## ##STR295## ##STR296## 556
##STR297## ##STR298## ##STR299## 556 ##STR300## ##STR301##
##STR302## 561 ##STR303## ##STR304## ##STR305## 572 ##STR306##
##STR307## ##STR308## 652 ##STR309## ##STR310## ##STR311## 555
##STR312## ##STR313## ##STR314## 672 ##STR315## ##STR316##
##STR317## 487 ##STR318## ##STR319## ##STR320## 600 ##STR321##
##STR322## ##STR323## 536 ##STR324## ##STR325## ##STR326## 554
##STR327## ##STR328## ##STR329## 534 ##STR330## ##STR331##
##STR332## 502 ##STR333## ##STR334## ##STR335## 574 ##STR336##
##STR337## ##STR338## 580 ##STR339## ##STR340## ##STR341## 581
##STR342## ##STR343## ##STR344## 533 ##STR345## ##STR346##
##STR347## 547 ##STR348## ##STR349## ##STR350## 548 ##STR351##
##STR352## ##STR353## 552 ##STR354## ##STR355## ##STR356## 539
##STR357## ##STR358## ##STR359## 584 ##STR360## ##STR361##
##STR362## 523 ##STR363## ##STR364## ##STR365## 568 ##STR366##
##STR367## ##STR368## 613 ##STR369## ##STR370## ##STR371## 602
##STR372## ##STR373## ##STR374## 572 ##STR375## ##STR376##
##STR377##
592 ##STR378## ##STR379## ##STR380## 637 ##STR381## ##STR382##
##STR383## 717 ##STR384## ##STR385## ##STR386## 651 ##STR387##
##STR388## ##STR389## 716 ##STR390## ##STR391## ##STR392## 671
##STR393## ##STR394## ##STR395## 524 ##STR396## ##STR397##
##STR398## 544 ##STR399## ##STR400## ##STR401## 506 ##STR402##
##STR403## ##STR404## 537 ##STR405## ##STR406## ##STR407## 648
##STR408## ##STR409## ##STR410## 581 ##STR411## ##STR412##
##STR413## 589 ##STR414## ##STR415## ##STR416## 537 ##STR417##
##STR418## ##STR419## 557 ##STR420## ##STR421## ##STR422## 616
##STR423## ##STR424## ##STR425## 636 ##STR426## ##STR427##
##STR428## 681 ##STR429## ##STR430## ##STR431## 522 ##STR432##
##STR433## ##STR434## 590 ##STR435## ##STR436## ##STR437## 624
##STR438## ##STR439## ##STR440## 534 ##STR441## ##STR442##
##STR443## 494 ##STR444## ##STR445## ##STR446## 550 ##STR447##
##STR448## ##STR449## 570 ##STR450## ##STR451## ##STR452## 624
##STR453## ##STR454## ##STR455## 674 ##STR456## ##STR457##
##STR458## 661 ##STR459## ##STR460## ##STR461## 654 ##STR462##
##STR463## ##STR464## 670 ##STR465## ##STR466## ##STR467## 680
##STR468## ##STR469## ##STR470## 636 ##STR471## ##STR472##
##STR473## 666 ##STR474## ##STR475## ##STR476## 520 ##STR477##
##STR478## ##STR479## 540 ##STR480## ##STR481## ##STR482## 598
##STR483## ##STR484## ##STR485## 643 ##STR486## ##STR487##
##STR488## 585 ##STR489## ##STR490## ##STR491## 601 ##STR492##
##STR493## ##STR494## 719 ##STR495## ##STR496## ##STR497## 594
##STR498## ##STR499## ##STR500## 686 ##STR501## ##STR502##
##STR503## 731 ##STR504## ##STR505## ##STR506## 573 ##STR507##
##STR508## ##STR509## 617 ##STR510## ##STR511## ##STR512## 492
##STR513## ##STR514## ##STR515## 725 ##STR516## ##STR517##
##STR518## 552 ##STR519## ##STR520## ##STR521## 743 ##STR522##
##STR523## ##STR524## 548 ##STR525## ##STR526## ##STR527## 568
##STR528## ##STR529## ##STR530## 613 ##STR531## ##STR532##
##STR533## 506 ##STR534## ##STR535## ##STR536## 780 ##STR537##
##STR538## ##STR539## 831 ##STR540## ##STR541## ##STR542## 676
##STR543## ##STR544## ##STR545## 524 ##STR546## ##STR547##
##STR548## 644 ##STR549## ##STR550## ##STR551## 689 ##STR552##
##STR553## ##STR554## 594 ##STR555## ##STR556## ##STR557## 681
##STR558## ##STR559## ##STR560## 726 ##STR561## ##STR562##
##STR563## 590 ##STR564## ##STR565## ##STR566## 594 ##STR567##
##STR568## ##STR569## 614 ##STR570## ##STR571## ##STR572## 659
##STR573## ##STR574## ##STR575## 534 ##STR576## ##STR577##
##STR578## 554 ##STR579## ##STR580## ##STR581## 599 ##STR582##
##STR583## ##STR584## 571 ##STR585## ##STR586## ##STR587## 648
##STR588## ##STR589## ##STR590## 643 ##STR591## ##STR592##
##STR593## 532 ##STR594## ##STR595## ##STR596## 552 ##STR597##
##STR598## ##STR599## 597 ##STR600## ##STR601## ##STR602## 602
##STR603## ##STR604## ##STR605## 562 ##STR606## ##STR607##
##STR608## 550 ##STR609## ##STR610## ##STR611## 532 ##STR612##
##STR613## ##STR614## 552 ##STR615## ##STR616## ##STR617## 528
##STR618## ##STR619## ##STR620## 573 ##STR621## ##STR622##
##STR623## 578 ##STR624## ##STR625## ##STR626## 516 ##STR627##
##STR628## ##STR629## 534 ##STR630## ##STR631## ##STR632## 520
##STR633## ##STR634## 536 ##STR635## ##STR636## ##STR637## 546
##STR638## ##STR639## ##STR640## 540 ##STR641## ##STR642##
##STR643## 560 ##STR644## ##STR645## ##STR646## 605 ##STR647##
##STR648## ##STR649## 562 ##STR650## ##STR651## ##STR652## 582
##STR653## ##STR654## ##STR655## 627 ##STR656## ##STR657##
##STR658## 508 ##STR659## ##STR660## ##STR661## 558 ##STR662##
##STR663## ##STR664## 522 ##STR665## ##STR666## ##STR667## 522
##STR668## ##STR669## ##STR670## 526 ##STR671## ##STR672##
##STR673## 591 ##STR674## ##STR675## ##STR676## 513 ##STR677##
##STR678## ##STR679## 585 ##STR680## ##STR681## ##STR682## 605
##STR683## ##STR684## ##STR685## 650 ##STR686## ##STR687##
##STR688## 617 ##STR689## ##STR690## ##STR691## 544 ##STR692##
##STR693## ##STR694## 589 ##STR695## ##STR696## ##STR697## 526
##STR698## ##STR699## ##STR700## 632 ##STR701## ##STR702##
##STR703## 616 ##STR704## ##STR705## ##STR706## 574 ##STR707##
##STR708## ##STR709## 585 ##STR710## ##STR711## ##STR712## 524
##STR713## ##STR714## ##STR715## 541 ##STR716## ##STR717##
##STR718## 542 ##STR719## ##STR720## ##STR721## 542 ##STR722##
##STR723## ##STR724## 607 ##STR725## ##STR726## ##STR727## 571
##STR728## ##STR729## ##STR730## 619 ##STR731## ##STR732##
##STR733## 560 ##STR734## ##STR735## ##STR736##
[0094] In another preferred embodiment of Formula I, M is
##STR737## In this embodiment, more preferred compounds are those
wherein A is .dbd.O, R is --(CH.sub.2).sub.n-- and X is --C(O)--. Y
is preferably chosen from --O--, --S--, --S(O)--, --S(O).sub.2--
and --NY.sup.4, and more preferably, is --O--.
[0095] Preferred compounds of this embodiment are those wherein W
is unsubstituted phenyl group or phenyl group having one or two
substituents chosen from lower alkyl group and halogen atom at the
ortho positions thereof. W.sup.1 is preferably unsubstituted
phenylene group or phenylene group having a substituent chosen from
methoxy group, lower alkyl group and halogen atom at the ortho
position to --NH--.
[0096] In this embodiment of Formula I, A is preferably .dbd.O and
A.sup.2 is a direct bond or --(CH.sub.2).sub.e--. More preferred
compounds are those wherein A.sup.2 is a direct bond and R.sup.8 is
--OH,
[0097] Preferred compounds of Formula I, wherein M is
##STR738##
[0098] and A is .dbd.O, are represented in Table 2. With respect to
the representation of --W.sup.1, the lower bond is the point of
attachment to --NH-- and the upper bond is the point of attachment
to --R--. The entry entitled --R--R.sup.8 depicts that portion of
the particular compound represented by TABLE-US-00002 TABLE 2
##STR739## Mass Spec (M.sup.+ + 1) W-- --W.sup.1-- -- --R-- --
--R.sup.8 534 ##STR740## ##STR741## ##STR742## 504 ##STR743##
##STR744## ##STR745## 524 ##STR746## ##STR747## ##STR748## 569
##STR749## ##STR750## ##STR751##
[0099] In another preferred embodiment of Formula I, M is
##STR752## In this embodiment, preferred compounds are those
wherein A is .dbd.O, R.sup.10 is --CO.sub.2H and q is 0 or 1. More
preferred are compounds wherein R.sup.10 is --CO.sub.2H, q is 0 or
1, most preferably 0, and m is 2.
[0100] When A is .dbd.O, L is preferably chosen from ##STR753##
##STR754##
[0101] More preferably, L is chosen from ##STR755##
[0102] Most preferably, L is chosen from ##STR756##
[0103] Preferred compounds of Formula I, wherein R is --CH.sub.2--
and X is .dbd.O, are those wherein W is unsubstituted phenyl group
or phenyl group having one or two substituents chosen from lower
alkyl group and halogen atom at the ortho positions thereof.
W.sup.1 is preferably unsubstituted phenylene group or phenylene
group having a substituent chosen from methoxy group, lower alkyl
group and halogen atom at the ortho position to --NH--.
[0104] Preferred compounds of Formula I, wherein M is
##STR757##
[0105] A is .dbd.O, R is --CH.sub.2-- and X is .dbd.O, are
represented in Table 3. With respect to the representation of
--W.sup.1, the lower bond is the point of attachment to --NH-- and
the upper bond is the point of attachment to --R--. The entry
entitled --N-Z.sup.2 depicts that portion of the particular
compound represented by TABLE-US-00003 TABLE 3 ##STR758## Mass Spec
(M.sup.+ + 1) W-- --W.sup.1-- --L-- --N-- --Z.sup.2 554.7
##STR759## ##STR760## ##STR761## ##STR762## 493.6 ##STR763##
##STR764## ##STR765## ##STR766## 540.6 ##STR767## ##STR768##
##STR769## ##STR770## 523.7 ##STR771## ##STR772## ##STR773##
##STR774## 555.7 ##STR775## ##STR776## ##STR777## ##STR778## 583.8
##STR779## ##STR780## ##STR781## ##STR782## 524.7 ##STR783##
##STR784## ##STR785## ##STR786## 509.7 ##STR787## ##STR788##
##STR789## ##STR790## 538.7 ##STR791## ##STR792## ##STR793##
##STR794## 554.7 ##STR795## ##STR796## ##STR797## ##STR798## 569.7
##STR799## ##STR800## ##STR801## ##STR802## 666.9 ##STR803##
##STR804## ##STR805## ##STR806## 567.8 ##STR807## ##STR808##
##STR809## ##STR810## 570.4 ##STR811## ##STR812## ##STR813##
##STR814## 552.7 ##STR815## ##STR816## ##STR817## ##STR818## 619.5
##STR819## ##STR820## ##STR821## ##STR822## 575.1 ##STR823##
##STR824## ##STR825## ##STR826## 553.6 ##STR827## ##STR828##
##STR829## ##STR830## 664.1 ##STR831## ##STR832## ##STR833##
##STR834## 629.7 ##STR835## ##STR836## ##STR837## ##STR838## 555.6
##STR839## ##STR840## ##STR841## ##STR842## 613.7 ##STR843##
##STR844## ##STR845## ##STR846## 555.7 ##STR847## ##STR848##
##STR849## ##STR850## 585.7 ##STR851## ##STR852## ##STR853##
##STR854## 591.7 ##STR855## ##STR856## ##STR857## ##STR858## 660.7
##STR859## ##STR860## ##STR861## ##STR862## 555.6 ##STR863##
##STR864## ##STR865## ##STR866## 556.6 ##STR867## ##STR868##
##STR869## ##STR870## 570.7 ##STR871## ##STR872## ##STR873##
##STR874## 570.7 ##STR875## ##STR876## ##STR877## ##STR878## 673.8
##STR879## ##STR880## ##STR881## ##STR882## 600.7 ##STR883##
##STR884## ##STR885## ##STR886## 552.7 ##STR887## ##STR888##
##STR889## ##STR890## 668.8 ##STR891## ##STR892## ##STR893##
##STR894## 583.7 ##STR895## ##STR896## ##STR897## ##STR898## 704.9
##STR899## ##STR900## ##STR901## ##STR902## 514.6 ##STR903##
##STR904## ##STR905## ##STR906## 514.6 ##STR907## ##STR908##
##STR909## ##STR910## 562.6 ##STR911## ##STR912## ##STR913##
##STR914##
[0106] Yet another preferred embodiment of Formula I includes
compounds wherein M is --R.sup.11-Z.sup.3-Q.sup.2-L.sup.1.
Preferably, A is .dbd.O, R is --CH.sub.2-- and X is .dbd.O.
Preferably W is unsubstituted phenyl group or phenyl group having
one or two substituents chosen from lower alkyl group and halogen
atom at the ortho positions thereof. W.sup.1 is preferably
unsubstituted phenylene group or phenylene group having a
substituent chosen from methoxy group, lower alkyl group and
halogen atom at the ortho position to --NH--.
[0107] In compounds wherein Q.sup.2 is ##STR915## and Z.sup.3 is a
divalent aliphatic hydrocarbon moiety, preferred compounds are
those wherein R.sup.11 is ##STR916## or --NR.sup.12, more
preferably NR.sup.12, wherein R.sup.12 is chosen from --H, lower
alkyl group and substituted lower alkyl group, most preferably
dihydroxy lower alkyl group. Preferred choices for Z.sup.3 is a
divalent aliphatic hydrocarbon moiety having 4, 5 or 6 carbon
atoms. A preferred choice for W.sup.1 is phenylene group having a
substituent chosen from methoxy group, lower alkyl group and
halogen atom at the ortho position to --NH--.
[0108] In compounds wherein Q.sup.2 is ##STR917## and Z.sup.3 is
##STR918## [0109] R.sup.11 is preferably --NR.sup.12--. In these
compounds, x and y are preferably 1. Preferred choices for R.sup.14
include --H, --OH and --F. A preferred choice for W.sup.1 is
phenylene group having a substituent chosen from methoxy group,
lower alkyl group and halogen atom at the ortho position to
--NH--.
[0110] In compounds wherein Q.sup.2 is ##STR919## and Z.sup.3 is
##STR920## R.sup.11 is preferably chosen from --O-- and
--NR.sup.12--, preferably wherein R.sup.12 is chosen from --H and
lower alkyl group. Preferably, R.sup.17 and R.sup.18 are each --H,
A preferred choice for W.sup.1 is phenylene group having a
substituent chosen from methoxy group, lower alkyl group and
halogen atom at the ortho position to --NH--.
[0111] In compounds wherein Q.sup.2 is ##STR921## and Z.sup.3 is
##STR922## R.sup.11 is preferably --NR.sup.12--, wherein R.sup.12
is preferably lower alkyl group. Preferred compounds of this
embodiment also include those wherein at least one of R.sup.17 and
R.sup.18 is lower alkyl group or substituted lower alkyl group.
[0112] In compounds wherein Q.sup.2 is ##STR923## and Z.sup.3 is
##STR924## R.sup.11 is preferably --NH-- and R.sup.17 and R.sup.18
are each preferably --H, A preferred choice for W.sup.1 is
phenylene group having a substituent chosen from methoxy group,
lower alkyl group and halogen atom at the ortho position to
--NH--.
[0113] In compounds wherein Q.sup.2 is chosen from aryl group,
substituted aryl group and ##STR925## and more preferably from
phenyl group and phenyl group substituted at the point of
attachment to Z.sup.3, Z.sup.3 is preferably a divalent aliphatic
hydrocarbon moiety.
[0114] Yet another embodiment of the invention is a compound
represented by Formula II, ##STR926## wherein the substituents W,
W.sup.1, R.sup.11 and Z.sup.3 are defined as in Formula I, and
L.sup.3 is chosen from ##STR927## wherein R.sup.20 is preferably
chosen from --H and lower alkyl, ##STR928##
[0115] Still another embodiment of the invention is a compound
represented by Formula III, ##STR929## wherein the substituents W,
W.sup.1, and R.sup.11 are defined as in Formula I, and d is chosen
from 0 and 1, and f is chosen from 1 and 2.
[0116] Preferred compounds of Formula I, wherein M is
-R.sup.11-Z.sup.3-Q.sup.2-L.sup.1, A is .dbd.O, R is --CH.sub.2--
and X is .dbd.O, are represented in Table 4. With respect to the
representation of --W.sup.1, the lower bond is the point of
attachment to --NH-- and the upper bond is the point of attachment
to --R--. The entry entitled --R.sup.11-L.sup.1 depicts that
portion of the particular compound represented by TABLE-US-00004
TABLE 4 ##STR930## Mass Spec (M.sup.+ + 1) W-- --W.sup.1-- --
--R.sup.11-- -- --L.sup.1 466.22 ##STR931## ##STR932## ##STR933##
496.22 ##STR934## ##STR935## ##STR936## 445.19 ##STR937##
##STR938## ##STR939## 475.20 ##STR940## ##STR941## ##STR942##
497.22 ##STR943## ##STR944## ##STR945## 510.42 ##STR946##
##STR947## ##STR948## 517.62 ##STR949## ##STR950## ##STR951##
473.56 ##STR952## ##STR953## ##STR954## 504.1 ##STR955## ##STR956##
##STR957## 529.17 ##STR958## ##STR959## ##STR960## 501.16
##STR961## ##STR962## ##STR963## 427.20 ##STR964## ##STR965##
##STR966## 441.22 ##STR967## ##STR968## ##STR969## 468.1 ##STR970##
##STR971## ##STR972## 532.2 ##STR973## ##STR974## ##STR975## 624.2
##STR976## ##STR977## ##STR978## 455.16 ##STR979## ##STR980##
##STR981## 483.70 ##STR982## ##STR983## ##STR984## 459.54
##STR985## ##STR986## ##STR987## 489.56 ##STR988## ##STR989##
##STR990## 487.59 ##STR991## ##STR992## ##STR993## 469.24
##STR994## ##STR995## ##STR996## 489.56 ##STR997## ##STR998##
##STR999## 489.56 ##STR1000## ##STR1001## ##STR1002## 459.54
##STR1003## ##STR1004## ##STR1005## 483.26 ##STR1006## ##STR1007##
##STR1008## 441.23 ##STR1009## ##STR1010## ##STR1011## 503.59
##STR1012## ##STR1013## ##STR1014## 477.53 ##STR1015## ##STR1016##
##STR1017## 507.55 ##STR1018## ##STR1019## ##STR1020## 521.58
##STR1021## ##STR1022## ##STR1023## ##STR1024## ##STR1025##
##STR1026## ##STR1027## ##STR1028## ##STR1029## ##STR1030##
##STR1031## ##STR1032## 414.10 ##STR1033## ##STR1034## ##STR1035##
441.22 ##STR1036## ##STR1037## ##STR1038## 511.47 ##STR1039##
##STR1040## ##STR1041## 459.11 ##STR1042## ##STR1043## ##STR1044##
504.6 ##STR1045## ##STR1046## ##STR1047## 500.2 ##STR1048##
##STR1049## ##STR1050## 530.2 ##STR1051## ##STR1052## ##STR1053##
516.4 ##STR1054## ##STR1055## ##STR1056## 486.2 ##STR1057##
##STR1058## ##STR1059## 544.2 ##STR1060## ##STR1061## ##STR1062##
500.2 ##STR1063## ##STR1064## ##STR1065## 506.2 ##STR1066##
##STR1067## ##STR1068## 516.2 ##STR1069## ##STR1070## ##STR1071##
498.2 ##STR1072## ##STR1073## ##STR1074## 475.3 ##STR1075##
##STR1076## ##STR1077## 502.2 ##STR1078## ##STR1079## ##STR1080##
506.3 ##STR1081## ##STR1082## ##STR1083## 518.1 ##STR1084##
##STR1085## ##STR1086## 530.1 ##STR1087## ##STR1088## ##STR1089##
526 ##STR1090## ##STR1091## ##STR1092## 602 ##STR1093## ##STR1094##
##STR1095## 573 ##STR1096## ##STR1097## ##STR1098## 622 ##STR1099##
##STR1100## ##STR1101## 646 ##STR1102## ##STR1103## ##STR1104## 518
##STR1105## ##STR1106## ##STR1107## 563 ##STR1108## ##STR1109##
##STR1110## 543 ##STR1111## ##STR1112## ##STR1113## 638 ##STR1114##
##STR1115## ##STR1116## 639 ##STR1117## ##STR1118## ##STR1119## 594
##STR1120## ##STR1121## ##STR1122## 593 ##STR1123## ##STR1124##
##STR1125## 513 ##STR1126## ##STR1127## ##STR1128## 638 ##STR1129##
##STR1130## ##STR1131## 593 ##STR1132## ##STR1133## ##STR1134## 624
##STR1135## ##STR1136## ##STR1137## 515 ##STR1138## ##STR1139##
##STR1140## 595 ##STR1141## ##STR1142## ##STR1143## 615 ##STR1144##
##STR1145## ##STR1146## 514 ##STR1147## ##STR1148## ##STR1149## 596
##STR1150## ##STR1151## ##STR1152## 616 ##STR1153## ##STR1154##
##STR1155## 516 ##STR1156## ##STR1157## ##STR1158## 486 ##STR1159##
##STR1160## ##STR1161## 521.56 ##STR1162## ##STR1163## ##STR1164##
535.55 ##STR1165## ##STR1166## ##STR1167## 507.54 ##STR1168##
##STR1169## ##STR1170## 535.59 ##STR1171## ##STR1172## ##STR1173##
522.51 ##STR1174## ##STR1175## ##STR1176## 539.55 ##STR1177##
##STR1178## ##STR1179## 502.56 ##STR1180## ##STR1181## ##STR1182##
521.56 ##STR1183## ##STR1184## ##STR1185## 547.6 ##STR1186##
##STR1187## ##STR1188## 620.69 ##STR1189## ##STR1190## ##STR1191##
633.73 ##STR1192## ##STR1193## ##STR1194## 590.67 ##STR1195##
##STR1196## ##STR1197## 495.5 ##STR1198## ##STR1199## ##STR1200##
529.94 ##STR1201## ##STR1202## ##STR1203## 491.54 ##STR1204##
##STR1205## ##STR1206## 525.98 ##STR1207## ##STR1208## ##STR1209##
489.56 ##STR1210## ##STR1211## ##STR1212## 461.51 ##STR1213##
##STR1214## ##STR1215## 495.95 ##STR1216## ##STR1217## ##STR1218##
491.54 ##STR1219## ##STR1220## ##STR1221## 461.51 ##STR1222##
##STR1223## ##STR1224## 495.95 ##STR1225## ##STR1226## ##STR1227##
525.98 ##STR1228## ##STR1229## ##STR1230## 534.6 ##STR1231##
##STR1232## ##STR1233## 538.57 ##STR1234## ##STR1235## ##STR1236##
504.58 ##STR1237## ##STR1238## ##STR1239## 552.59 ##STR1240##
##STR1241## ##STR1242## 523.55 ##STR1243## ##STR1244## ##STR1245##
529.94 ##STR1246## ##STR1247## ##STR1248## 490.55 ##STR1249##
##STR1250## ##STR1251## 543.97 ##STR1252## ##STR1253## ##STR1254##
540.01 ##STR1255## ##STR1256## ##STR1257## 548.63 ##STR1258##
##STR1259## ##STR1260## 504.58 ##STR1261## ##STR1262## ##STR1263##
620.07 ##STR1264## ##STR1265## ##STR1266## 562.66 ##STR1267##
##STR1268## ##STR1269## 578.63 ##STR1270## ##STR1271## ##STR1272##
506.55 ##STR1273## ##STR1274## ##STR1275## 574.67 ##STR1276##
##STR1277## ##STR1278## 510.51 ##STR1279## ##STR1280## ##STR1281##
506.55 ##STR1282## ##STR1283## ##STR1284##
505.56 ##STR1285## ##STR1286## ##STR1287## 556.41 ##STR1288##
##STR1289## ##STR1290## 596.67 ##STR1291## ##STR1292## ##STR1293##
532.63 ##STR1294## ##STR1295## ##STR1296## 691.53 ##STR1297##
##STR1298## ##STR1299## 541.0 ##STR1300## ##STR1301## ##STR1302##
607 ##STR1303## ##STR1304## ##STR1305## 497 ##STR1306## ##STR1307##
##STR1308## 525 ##STR1309## ##STR1310## ##STR1311## 527 ##STR1312##
##STR1313## ##STR1314## 542 ##STR1315## ##STR1316## ##STR1317## 498
##STR1318## ##STR1319## ##STR1320## 543 ##STR1321## ##STR1322##
##STR1323## 611 ##STR1324## ##STR1325## ##STR1326## 477 ##STR1327##
##STR1328## ##STR1329## 512 ##STR1330## ##STR1331## ##STR1332## 518
##STR1333## ##STR1334## ##STR1335## 569 ##STR1336## ##STR1337##
##STR1338## 569 ##STR1339## ##STR1340## ##STR1341## 592 ##STR1342##
##STR1343## ##STR1344## 591 ##STR1345## ##STR1346## ##STR1347## 569
##STR1348## ##STR1349## ##STR1350## 538 ##STR1351## ##STR1352##
##STR1353## 583 ##STR1354## ##STR1355## ##STR1356## 561 ##STR1357##
##STR1358## ##STR1359## 549 ##STR1360## ##STR1361## ##STR1362## 604
##STR1363## ##STR1364## ##STR1365## 589 ##STR1366## ##STR1367##
##STR1368## 506 ##STR1369## ##STR1370## ##STR1371## 607 ##STR1372##
##STR1373## ##STR1374## 589 ##STR1375## ##STR1376## ##STR1377## 575
##STR1378## ##STR1379## ##STR1380## 619 ##STR1381## ##STR1382##
##STR1383## 601 ##STR1384## ##STR1385## ##STR1386## 603 ##STR1387##
##STR1388## ##STR1389## 617 ##STR1390## ##STR1391## ##STR1392## 625
##STR1393## ##STR1394## ##STR1395## 611 ##STR1396## ##STR1397##
##STR1398## 522 ##STR1399## ##STR1400## ##STR1401## 565 ##STR1402##
##STR1403## ##STR1404## 593 ##STR1405## ##STR1406## ##STR1407## 518
##STR1408## ##STR1409## ##STR1410## 538 ##STR1411## ##STR1412##
##STR1413## 627 ##STR1414## ##STR1415## ##STR1416## 585 ##STR1417##
##STR1418## ##STR1419## 624 ##STR1420## ##STR1421## ##STR1422## 609
##STR1423## ##STR1424## ##STR1425## 639 ##STR1426## ##STR1427##
##STR1428## 623 ##STR1429## ##STR1430## ##STR1431## 637 ##STR1432##
##STR1433## ##STR1434## 595 ##STR1435## ##STR1436## ##STR1437## 621
##STR1438## ##STR1439## ##STR1440## 645 ##STR1441## ##STR1442##
##STR1443## 581 ##STR1444## ##STR1445## ##STR1446## 637 ##STR1447##
##STR1448## ##STR1449## 611 ##STR1450## ##STR1451## ##STR1452## 631
##STR1453## ##STR1454## ##STR1455## 488 ##STR1456## ##STR1457##
##STR1458## 504 ##STR1459## ##STR1460## ##STR1461## 522 ##STR1462##
##STR1463## ##STR1464## 640 ##STR1465## ##STR1466## ##STR1467## 676
##STR1468## ##STR1469## ##STR1470## 548 ##STR1471## ##STR1472##
##STR1473## 613 ##STR1474## ##STR1475## ##STR1476## 666 ##STR1477##
##STR1478## ##STR1479## 614 ##STR1480## ##STR1481## ##STR1482## 626
##STR1483## ##STR1484## ##STR1485## 630 ##STR1486## ##STR1487##
##STR1488## 676 ##STR1489## ##STR1490## ##STR1491## 506 ##STR1492##
##STR1493## ##STR1494## 526 ##STR1495## ##STR1496## ##STR1497## 502
##STR1498## ##STR1499## ##STR1500## 514 ##STR1501## ##STR1502##
##STR1503## 484 ##STR1504## ##STR1505## ##STR1506## 579 ##STR1507##
##STR1508## ##STR1509## 585 ##STR1510## ##STR1511## ##STR1512## 512
##STR1513## ##STR1514## ##STR1515## 482 ##STR1516## ##STR1517##
##STR1518## 532 ##STR1519## ##STR1520## ##STR1521## 560 ##STR1522##
##STR1523## ##STR1524## 625 ##STR1525## ##STR1526## ##STR1527## 583
##STR1528## ##STR1529## ##STR1530## 568 ##STR1531## ##STR1532##
##STR1533## 534 ##STR1534## ##STR1535## ##STR1536## 574 ##STR1537##
##STR1538## ##STR1539## 536 ##STR1540## ##STR1541## ##STR1542## 581
##STR1543## ##STR1544## ##STR1545## 522 ##STR1546## ##STR1547##
##STR1548## 520 ##STR1549## ##STR1550## ##STR1551##
[0117] The principles of the present invention also encompass
prodrugs in the scope of Formula I, and compounds representative
thereof include those wherein R.sup.5 or R.sup.8 is a lower alkoxy
group, and those wherein R.sup.10 or R.sup.19 is a lower
alkoxycarbonyl group.
[0118] The principles of the present invention also provide a
method for inhibiting cell adhesion, and in particular, VLA-4
mediated cell adhesion at .alpha.4.beta.1 receptor sites in a
mammal, wherein the method comprises administering an effective
amount of a compound represented by Formula I. As used herein,
inhibiting cell adhesion is intended to include inhibiting,
suppressing and preventing VLA-4 mediated cell adhesion-associated
conditions, including but not limited to, inflammation and cell
adhesion-associated immune or autoimmune responses.
[0119] The principles of the present invention therefore also
provide a method of treating a condition associated with VLA-4
mediated cell adhesion, wherein the method comprises administering
to a mammal in need of such treatment, an effective amount of a
compound represented by Formula I. Such conditions include for
example, but are not limited to, inflammatory and autoimmune
responses, diabetes, asthma, arthritis, psoriasis, multiple
sclerosis, inflammatory bowel disease, transplantation rejection,
and tumor metastasis. As used herein, "treatment" of a mammal is
intended to include prophylaxis as well.
[0120] The compounds of the present invention may be administered
as a monotherapy, or in combination with antiinflammatory or
immunosuppressive agents. Such combination therapies can involve
the administration of the various pharmaceuticals as a single
dosage form or as multiple dosage forms administered at the same
time or at different times.
[0121] Any suitable route of administration may be employed for
providing a patient with an effective amount of a compound of the
present invention. Suitable routes of administration may include,
for example, oral, rectal, nasal, buccal, parenteral (such as,
intravenous, intrathecal, subcutaneous, intramuscular,
intrasternal, intrahepatic, intralesional, intracranial,
intra-articular, and intra-synovial), transdermal (such as, for
example, patches), and the like. Due to their ease of
administration, oral dosage forms, such as, for example, tablets,
troches, dispersions, suspensions, solutions, capsules, soft
gelatin capsules, and the like, may be preferred. Administration
may also be by controlled or sustained release means and delivery
devices. Methods for the preparation of such dosage forms are well
known in the art.
[0122] Pharmaceutical compositions incorporating compounds of the
present invention may include excipients, a pharmaceutically
acceptable carrier, in addition to other therapeutic ingredients.
Excipients such as starches, sugars, microcrystalline cellulose,
diluents, lubricants, binders, coloring agents, flavoring agents,
granulating agents, disintegrating agents, and the like may be
appropriate depending upon the route of administration. Because of
their ease of administration, tablets and capsules represent the
most advantageous oral dosage unit forms. If desired, tablets may
be coated by standard aqueous or nonaqueous techniques.
[0123] The compounds of the present invention may be used in the
form of pharmaceutically acceptable salts derived from inorganic or
organic bases. Suitable pharmaceutically acceptable base addition
salts include, but are not limited to, ammonium salts, alkali metal
salts, metallic salts made from aluminum, calcium, lithium,
magnesium, potassium, sodium and zinc, organic salts made from
chloroprocaine, choline, N,N'-dibenzylethylenediamine,
dicyclohexylamine, diethanolamine, ethylenediamine, lysine,
meglumine (N-methylglucamine) and procaine, as well as salts with
amino acids, such as arginine, lysine, and so forth.
[0124] Where the compounds of the invention have a basic moiety,
such as an amino group, the compounds may be used in the form of
pharmaceutically acceptable non-toxic organic or inorganic acids.
Such acids include acetic, benzenesulfonic, benzoic,
camphorsulfonic, citric, ethanesulfonic, methanesulfonic, fumaric,
gluconic, glutamic, hydrobromic, hydrochloric, lactic, maleic,
malic, mandelic, nitric, pamoic, pantothenic, phosphoric, succinic,
sulfuric, tartaric, p-toluenesulfonic acids, and the like.
Particularly preferred are citric, hydrochloric, maleic, fumaric,
phosphoric, sulfuric, tartaric and p-toluenesulfonic acids.
Compounds of the invention may also be in the form of hydrates.
DETAILED DESCRIPTION OF THE INVENTION
Abbreviations & Definitions
[0125] The following terms and abbreviations have the indicated
meaning throughout this disclosure. [0126] 293E HEK cells=293E
human embryonic kidney cells [0127] Ac=acetyl [0128]
anti-.alpha.4-PE conjugated=monoclonal antibody against integrin
.alpha.4 subunit, phycoerythrin conjugated [0129] anti-.beta.1-FITC
conjugated=monoclonal antibody against integrin .beta.1 subunit,
fluorescein conjugated [0130] .alpha.5.beta.1=integrin
.alpha.5.beta.1, fibronectin receptor, VLA-5 [0131]
.alpha.4.beta.3=integrin .alpha.v.beta.3, vitronectin receptor
[0132] .alpha.4.beta.7=integrin .alpha.4.beta.7 [0133] Bn=benzyl
[0134] Boc=t-butoxycarbonyl [0135] BSA=bovine serum albumin [0136]
c-=cyclo- [0137] cDNA=complementary DNA [0138] CHO cells=Chinese
Hamster Ovary cells [0139] p-ClPh=para-chlorophenyl [0140] CMV
promoter=cytomegalovirus promoter [0141]
m-CPBA=3-chloroperoxybenzoic acid [0142] DAST=diethylaminosulfur
trifluoride [0143] DCM=dichloromethane=methylene
chloride=CH.sub.2Cl.sub.2 [0144] DELFIA=dissociation enhanced
lanthanide fluor-immune assay [0145] DIAD=diisopropyl
azodicarboxylate [0146] DIC=diisopropylcarbodiimide [0147]
DIEA=N,N-diisopropylethylamine [0148]
DMAP=4-N,N-dimethylaminopyridine [0149] DMEM=Dulbecco's Modified
Eagle's Medium [0150] DMF=N,N-dimethylformamide [0151]
DTPA=Diethylenetriaminepentaacetic acid [0152]
EDC=1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [0153]
Et.sub.2O=ethyl ether [0154] FACS fluorescence cell sorting [0155]
Fmoc=9-fluorenylmethoxycarbonyl [0156] GPIIb/IIIa=integrin
.alpha.II.beta.3, fibrinogen receptor [0157]
HEPES=N-(2-hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid)
[0158] HMDS=1,1,1,3,3,3-hexamethyldisilzane [0159] HOAc=acetic acid
[0160] HOBt=1-hydroxybenzotriazole [0161] human IgG1=human
immunoglobulin G1 [0162] ICAM=intracellular adhesion molecule
[0163] LDV=Leu-Asp-Val [0164] LFA-1 and Mac-1=Lymphocyte
function-related antigen [0165] LiHMDS=lithium
1,1,1,3,3,3-hexamethyldisilazane [0166] Me=methyl [0167]
p-MeOPh=para-methoxyphenyl [0168] nM=nanomolar [0169] PBS=phosphate
buffered saline [0170] PEG=polyethylene glycol [0171] Ph=phenyl
[0172] PhOH=phenol [0173] PyBroP=bromo-tris-pyrrolidinphosphonium
hexafluorophosphate [0174] RPMI medium=Russell Park Memorial
Institute medium [0175] TFA=trifluoroacetic acid [0176]
TFAA=trifluoroacetic acid anhydride [0177] THF=tetrahydrofuran
[0178] TLC=thin-layer chromatography [0179] TMS=trimethylsilyl
[0180] Ts=toluenesulfonyl [0181] VCAM-1 (D1D7)=vascular cell
adhesion molecule (containing one to seven immuloglobulin domains)
[0182] VCAM-IgG fusion protein a VCAM IgG fusion protein containing
the one to seven immunoglobulin domains of human VCAM-1 (D1D7)
attached above the hinge region of an IgG1 molecule
[0183] "Alkyl group" is intended to include linear or branched
hydrocarbon radicals and combinations thereof of 1 to 20 carbons.
"Lower alkyl group" means alkyl groups of from 1 to about 10,
preferably from 1 to about 8, and more preferably, from 1 to about
6 carbon atoms. Examples of such radicals include methyl, ethyl,
n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl,
pentyl, iso-amyl, hexyl, octyl groups and the like.
[0184] "Alkylene group" means a divalent radical formed by removing
a hydrogen atom from an "alkyl group."
[0185] "Aryl group" means a radical formed from an aromatic
hydrocarbon ring of 4 to about 16 carbon atoms, preferably of 6 to
about 12 carbon atoms, and more preferably of 6 to about 10 carbon
atoms. The rings may optionally be substituted with 1-3
substituents selected from alkyl, halogen, hydroxy, alkoxy,
aryloxy, haloalkyl, phenyl and heteroaryl. Examples of aryl groups
are phenyl, biphenyl, 3,4-dichlorophenyl and naphthyl.
[0186] "Arylene group" means a divalent radical formed by removing
a hydrogen atom from an "aryl group."
[0187] "Arylalkyl group" denotes a structure comprising an alkyl
attached to an aryl ring. Examples include benzyl, phenethyl,
4-chlorobenzyl, and the like.
[0188] "Cycloalkyl group" refers to a saturated hydrocarbon ring
radical of from 3 to 12 carbon atoms, and preferably from 3 to 8
carbon atoms. Examples include cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, norbornyl, adamantyl, myrtanyl groups and
the like. "Lower cycloalkyl group" refers to cycloalkyl of 3 to 6
carbons.
[0189] "Cycloalkylene group" means a divalent radical formed by
removing a hydrogen atom from a "cycloalkyl group."
[0190] "Divalent C.sub.1 to C.sub.20 aliphatic hydrocarbon-moiety"
includes alkylene, cycloalkylene, alkenylene, alkynylene groups and
combinations thereof. Examples include ethylene, propylene,
propynylene, 2,4-heptadienylene groups and the like.
[0191] "Heterocyclyl group" refers to a cyclic radical having from
1 to 6 carbon atoms, preferably 3 to 6 carbon atoms, and from 1 to
4 heteroatoms chosen from O, N and S. Examples include: pyrrolyl,
pyridinyl, pyrazolyl, triazolyl, pyrimidinyl, pyridazinyl,
oxazolyl, thiazolyl, imidazolyl, indolyl, thienyl, furyl,
azetidiyl, tetrazolyl, 2-pyrrolinyl, 3-pyrrolinyl, pyrrolindinyl,
1,3-dioxolanyl, imidazolinyl, imidazolidinyl, pyrazolinyl,
pyrazolidinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl,
1,2,3-triazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4H-pyranyl,
piperidinyl, 1,4-dithianyl, morpholinyl, thiomorpholinyl,
pyrazinyl, piperazinyl, 1,3,5-triazinyl, 1,2,5-trithianyl,
benzo(b)thiophenyl, benzimidazolyl, quinolinyl groups and the
like.
[0192] "Heterocyclylene group" means a radical formed by removing a
hydrogen atom from a "heterocyclyl group."
[0193] "Heteroaryl group" refers to an aromatic cyclic radical
having from 1 to 12 carbon atoms, preferably 1 to 6 carbon atoms,
and from 1 to 4 heteroatoms chosen from O, N and S; or a bicyclic
9- or 10-membered heteroaromatic ring system containing 14
heteroatoms selected from O, N and S. The methine H atoms of a
heterocyclyl or heteroaryl structure may be optionally substituted
with alkyl, alkoxy or halogen. Examples include: imidazolyl,
pyridyl, indolyl, thienyl, benzopyranyl, thiazolyl, furyl,
benzimidazolyl, quinolinyl, isoquinolinyl, quinoxalinyl,
pyrimidinyl, pyrazinyl, tetrazolyl, pyrazolyl groups and the
like.
[0194] "Heteroarylene group" means a divalent radical formed by
removing a hydrogen atom from a "heteroaryl group."
[0195] "Alkoxy group" means a straight, branched or cyclic
hydrocarbon configuration and combinations thereof, including from
1 to 20 carbon atoms, preferably from 1 to 8 carbon atoms, more
preferably from 1 to 4 carbon atoms, and an oxygen atom at the
point of attachment. Suitable alkoxy groups include methoxy,
ethoxy, n-propoxy, isopropoxy, n-butoxy, iso-butoxy, sec-butoxy,
tert-butoxy, cyclopropoxy, cyclohexyloxy groups and the like.
"Lower alkoxy group" refers to alkoxy groups having from 1 to 4
carbon atoms.
[0196] "Alkenyl group" refers to an unsaturated acyclic hydrocarbon
radical in so much as it contains at least one double bond. "Lower
alkenyl group" refers to such radicals containing from 2 to 10
carbon atoms, preferably from 2 to 8 carbon atoms and more
preferably from 2 to 6 carbon atoms. Examples of suitable alkenyl
radicals include propenyl, buten-1-yl, isobutenyl, penten-1-yl,
2-methylbuten-1-yl, 3-methylbuten-1-yl, hexen-1-yl, hepten-1-yl,
and octen-1-yl groups and the like.
[0197] "Alkenylene group" means a divalent radical formed by
removing a hydrogen atom from an "alkenyl group."
[0198] "Alkynyl group" refers to an unsaturated acyclic hydrocarbon
radical containing at least one triple bond. Examples include
ethynyl, propynyl groups, and the like.
[0199] "Alkynylene group" means a divalent radical formed by
removing a hydrogen atom from an alkynyl group."
[0200] "Substituted alkyl group" means a linear or branched alkyl
group wherein at least one hydrogen atom attached to an aliphatic
carbon is replaced with a substituent such as alkyl, amino, alkoxy,
hydroxy, aryl, cyano, carboxy, alkoxycarbonyl, monoalkylamino,
alkyloxy, cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl,
alkylsulfonyl, arylthio, carboxyalkyl, alkoxycarbonylalkyl,
haloalkyl, acylamino, dialkylamino, cyclicamino groups, halogen
atom and nitro. Examples of such substituent groups include methyl,
isopropyl, methoxy, ethoxy, propoxy, amino, methylamino, phenyl,
naphthyl groups, chlorine, fluorine and the like.
[0201] "Substituted alkylene group" means a linear or branched
alkylene group wherein at least one hydrogen atom attached to an
aliphatic carbon is replaced with a substituent such as alkyl,
amino, alkoxy, hydroxy, aryl, cyano, carboxy, alkoxycarbonyl,
monoalkylamino, alkyloxy, cyanoalkyl, cycloalkyl, alkylthio,
alkylsulfinyl, alkylsulfonyl, arylthio, carboxyalkyl,
alkoxycarbonylalkyl, haloalkyl, acylamino, dialkylamino,
cyclicamino groups, halogen atom and nitro.
[0202] "Substituted cycloalkyl group" means a cycloalkyl group
wherein at least one hydrogen atom attached to a ring carbon atom
is replaced with a substituent such as alkyl, amino, alkoxy,
hydroxy, aryl, cyano, carboxy, alkoxycarbonyl, monoalkylamino,
alkyloxy, cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl,
alkylsulfonyl, arylthio, carboxyalkyl, alkoxycarbonylalkyl,
haloalkyl, acylamino, dialkylamino, cyclicamino groups, halogen
atom and nitro.
[0203] "Substituted cycloalkyene group" means a cycloalkylene group
wherein at least one hydrogen atom attached to a ring carbon is
replaced with a substituent such as alkyl, amino, alkoxy, hydroxy,
aryl, cyano, carboxy, alkoxycarbonyl, monoalkylamino, alkyloxy,
cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl,
arylthio, carboxyalkyl, alkoxycarbonylalkyl, haloalkyl, acylamino,
dialkylamino, cyclicamino groups, halogen atom and nitro.
[0204] "Substituted aryl group" means an aryl group wherein at
least one methine hydrogen atom attached to an aromatic carbon is
replaced with a substituent such as alkyl, amino, alkoxy, hydroxy,
aryl, cyano, carboxy, alkoxycarbonyl, monoalkylamino, alkyloxy,
cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl,
arylthio, carboxyalkyl, alkoxycarbonylalkyl, haloalkyl, acylamino,
dialkylamino, cyclicamino groups, halogen atom and nitro.
[0205] Substituted arylene group" means an arylene group wherein at
least one hydrogen atom attached to an aromatic carbon is replaced
with a substituent such as alkyl, amino, alkoxy, hydroxy, aryl,
cyano, carboxy, alkoxycarbonyl, monoalkylamino, alkyloxy,
cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl,
arylthio, carboxyalkyl, alkoxycarbonylalkyl, haloalkyl, acylamino,
dialkylamino, cyclicamino groups, halogen atom and nitro.
[0206] "Substituted heteroaryl group" or "substituted heterocyclyl
group" means a heteroaryl or heterocyclyl group wherein at least
one hydrogen atom attached to a ring thereof is replaced with a
substituent such as alkyl, amino, alkoxy, hydroxy, aryl, cyano,
carboxy, alkoxycarbonyl, monoalkylamino, alkyloxy, cyanoalkyl,
cycloalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl, arylthio,
carboxyalkyl, alkoxycarbonylalkyl, haloalkyl, acylamino,
dialkylamino, cyclicamino groups, halogen atom and nitro.
[0207] "Substituted heteroarylene group" or "substituted
heterocyclylene group" means a heteroarylene or heterocyclylene
group wherein at least one hydrogen atom attached to a ring thereof
is replaced with a substituent such as alkyl, amino, alkoxy,
hydroxy, aryl, cyano, carboxy, alkoxycarbonyl, monoalkylamino,
alkyloxy, cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl,
alkylsulfonyl, arylthio, carboxyalkyl, alkoxycarbonylalkyl,
haloalkyl, acylamino, dialkylamino, cyclicamino groups, halogen
atom and nitro.
[0208] "Substituted arylalkyl group" means an arylalkyl having one
or more substituents such as alkyl, amino, alkoxy, hydroxy, aryl,
cyano, carboxy, alkoxycarbonyl, monoalkylamino, alkyloxy,
cyanoalkyl, cycloalkyl, alkylthio, alkylsulfinyl, alkylsulfonyl,
arylthio, carboxyalkyl, haloalkyl, alkoxycarbonylalkyl, acylamino,
dialkylamino, cyclicamino groups, halogen atom and nitro.
[0209] "Halogen" is intended to include for example, F, Cl, Br and
I.
[0210] The term "prodrug" refers to a chemical compound that is
converted to an active agent by metabolic processes in vivo. [See,
e.g., N. Boder and J. J. Kaminski, Ann. Rep. Med. Chem. 22:303
(1987) and H, Bundgarrd, Adv. Drug Delivery Rev., 3:39 (1989)]. The
use of prodrug precursors of compounds of the present invention in
any of the methods described herein is contemplated and is intended
to be within the scope of the invention.
[0211] Terminology related to "protected," "protecting" and/or
"deprotecting" functionalities is used throughout this application.
Such terminology is well understood by persons of skill in the art
and is used in the context of processes which involve sequential
treatment with a series of reagents. In this context, a protecting
group refers to a group which is used to mask a functionality
during a process step in which it would otherwise react, but in
which reaction is undesirable. The protecting group prevents
reaction at that step, but may be subsequently removed to expose
the original functionality. The removal or "deprotection" occurs
after the completion of the reaction or reactions in which the
functionality would interfere. Thus, when a sequence of reagents is
specified, as it is in the processes of the invention, the person
of ordinary skill can readily envision those groups that would be
suitable as "protecting groups" for the functionalities
involved.
[0212] In the case of the present invention, the functionalities
that must be protected are amines. Suitable groups for that purpose
are discussed in standard textbooks in the field of chemistry, such
as Protective Groups in Organic Synthesis by T. W. Greene [John
Wiley & Sons, New York, 1991], which is incorporated herein by
reference. Particular attention is drawn to the chapter entitled
"Protection for the Amino Group" (pages 309-405). Preferred
protecting groups include BOC and Fmoc. Exemplary methods for
protecting and deprotecting with these groups are found in Greene
and Wuts on pages 318 and 327.
[0213] The materials upon which the syntheses described herein are
performed are referred to as solid supports, beads, and resins.
These terms are intended to include: (a) beads, pellets, disks,
fibers, gels, or particles such as cellulose beads, pore-glass
beads, silica gels, polystyrene beads optionally cross-linked with
divinylbenzene and optionally grafted with polyethylene glycol,
poly-acrylamide beads, latex beads, dimethylacrylamide beads
optionally cross-linked with N,N'-bis-acryloyl ethylene diamine,
glass particles coated with hydrophobic polymer, etc., i.e.,
material having a rigid or semi-rigid surface; and (b) soluble
supports such as polyethylene glycol or low molecular weight,
non-cross-linked polystyrene. The solid supports may, and usually
do, have functional groups such as amino, hydroxy, carboxy, or halo
groups; where amino groups are the most common.
[0214] Tentagel.TM. NH.sub.2 (Rapp Polymere, Tubingen, Germany) is
a preferred amine functionalized polyethylene glycol-grafted
polystyrene resin. Tentagel.TM.-S-PHB resin has a para-hydroxy
benzyl linker which can be cleaved by the use of 90%
trifluoroacetic acid in dichloromethane. Techniques for
functionalizing the surface of solid phases are well known in the
art. Attachment of lysine to the amino groups on a bead (to
increase the number of available sites) and subsequent attachment
of linkers as well as further steps in a typical combinatorial
synthesis are described, for example, in PCT application
WO95/30642, the disclosure of which is incorporated herein by
reference. In the synthesis described in WO95/30642, the linker is
a photolytically cleavable linker, but the general principles of
the use of a linker are well illustrated.
Optical Isomers--Diastereomers--Geometric Isomers
[0215] Some of the compounds described herein contain one or more
asymmetric centers and may thus give rise to enantiomers,
diastereomers, and other stereoisometric forms which may be defined
in terms of absolute stereochemistry as (R)- or (S)-, or as (D)- or
(L)- for amino acids. The present invention is meant to include all
such possible diastereomers as well as their racemic and optically
pure forms. Optically active (R)- and (S)-, or (D)- and (L)-isomers
may be prepared using chiral synthons or chiral reagents, or
resolved using conventional techniques. When the compounds
described herein contain olefinic double bonds or other centers of
geometric asymmetry, and unless specified otherwise, it is intended
to include both (E)- and (Z)-geometric isomers. Likewise, all
tautomeric forms are intended to be included. The configuration of
any carbon-carbon double bond appearing herein is selected for
convenience only and is not intended to designate a particular
configuration; thus a carbon-carbon double bond depicted
arbitrarily herein as trans may be cis, trans, or a mixture of the
two in any proportion.
[0216] In view of the above definitions, other chemical terms used
throughout this application can be easily understood by those of
skill in the art. Terms may be used alone or in any combination
thereof. The preferred and more preferred chain lengths of the
radicals apply to all such combinations.
Utility
[0217] The compounds of the present invention have demonstrated
utility as selective inhibitors at VLA-4 receptors. The inhibitory
concentration (IC.sub.50) and the VLA-4 selectivity of test
compounds for an .alpha.4.beta.1 receptor using in vitro assays are
determined in direct binding assays and competitive assays with
other integrin receptors such as .beta.2 (LFA-1 and Mac-1), .beta.3
(GPIIb/IIIa and .alpha.v.beta.3) and .beta.1 (.alpha.4.beta.7).
Compounds of the present invention have K.sub.i values <1 .mu.M.
Preferred compounds of the invention are those having K.sub.i
values <300 nM, more preferably <100 nM, even more preferably
<50 nM, and most preferably, <12 nM.
[0218] Examples of preferred compounds having a K.sub.i value
<50 nM are shown below. These examples are provided by way of
illustration only, and are not intended to limit the invention
thereto. ##STR1552## ##STR1553## ##STR1554## ##STR1555##
##STR1556## ##STR1557## ##STR1558## ##STR1559## ##STR1560##
##STR1561## ##STR1562## ##STR1563## ##STR1564## ##STR1565##
##STR1566## ##STR1567## ##STR1568## ##STR1569## ##STR1570##
##STR1571## ##STR1572## ##STR1573## ##STR1574## ##STR1575##
##STR1576## ##STR1577## ##STR1578## ##STR1579## ##STR1580##
##STR1581## ##STR1582## ##STR1583## ##STR1584## ##STR1585##
##STR1586## ##STR1587## ##STR1588## ##STR1589## ##STR1590##
##STR1591## ##STR1592## ##STR1593## ##STR1594## ##STR1595##
##STR1596## ##STR1597## ##STR1598## ##STR1599## ##STR1600##
##STR1601## ##STR1602## ##STR1603## ##STR1604## ##STR1605##
##STR1606## ##STR1607## ##STR1608## ##STR1609## ##STR1610##
##STR1611## ##STR1612## ##STR1613## ##STR1614## ##STR1615##
In Vitro Assays
[0219] A direct binding assay was used to quantify the inhibitory
activity of the compounds. In this assay, VLA-4-expressing cells
were seeded in a 96-well microtiter plate. The cells were allowed
to grow for 2 days until confluent. Various concentrations of the
test compound were added together with 2 nM of the
europium-labeled, VCAM-IgG fusion protein. The cells were allowed
to incubate at room temperature in the microwells for at least 30
minutes. Following incubation, the microwells were emptied and
washed. The amount of europium-labeled VCAM-IgG fusion protein
bound was determined by time-resolved fluorescence measurement
Inhibition of binding was determined by quantifying the
fluorescence bound to the plate for each of the various
concentrations of test compound, as well as for controls containing
no test compound.
[0220] The VLA-4-expressing cells used in this assay was a CHO cell
line stably transfected with the cDNA of the human .alpha.4 and
.beta.1 subunits. Construction and maintenance of the cell line are
described in the assay procedures. A VCAM IgG fusion protein
containing the one to seven immunoglobulin domains of human VCAM-1
(D1D7) attached above the hinge region of an IgG1 molecule was
labeled with europium chelates. The preparation and labeling of the
fusion protein are described in the assay procedures.
[0221] The cell adhesion inhibitory activity of the test compound
was determined by blocking the Jurkat cell attachment to the
D1D7-VCAM IgG fusion protein. Jurkat cell is a human lymphocytic
cell line expressing VLA-4 on cell surface. In this assay, each of
the 96-well microtiter wells was coated with 75 ng of the VCAM IgG
fusion protein. The wells were then blocked by the addition of 1%
bovine serum albumin to remove nonspecific adhesive sites. Varying
concentrations of the test compound were added together with the
calcein-labeled Jurkat cells. The cells were allowed to adhere to
the VCAM coated wells at room temperature for 1 hour in the dark.
Following incubation, the plate was washed by immersing face down
into a container filled with phosphate buffered saline. The wells
were blotted dry on paper towel. Quantitation of the adhered cell
was determined by fluorescence measurement. Decreased fluorescence
indicated inhibition of cell adhesion by the test compound.
[0222] Specificity for .alpha.4.beta.1 of each test compound among
other integrin receptors, namely, .beta.2 (LFA-1 and Mac-1),
.beta.3 (GPIIb/IIIa and .alpha.v.beta.3), .beta.1 (.alpha.5.beta.1)
and .beta.7 (.alpha.4.beta.7) was examined. LFA-1 binds to ICAM-1
and mediates the emigration of leukocytes into inflammatory sites.
Mac-1 binds to a number of ligands, including ICAM-1 and
fibrinogen, and plays an important role in neutrophil phagocytosis
and oxygen free radical generation. GPIIb/IIIa on platelet surface
binds to fibrinogen in plasma and induces platelet aggregation.
.alpha.v.beta.3 binds to a number of extracellular matrix proteins,
including vitronectin and mediates cell migration and prevents cell
apoptosis. .alpha.4.beta.7 shares the same ligands as VLA-4
(VCAM-1, MAdCAM-1, and fibronectin), but with different preference.
This receptor is expressed on lymphoid cells and is involved in
lymphocyte migration to mucosal tissues.
[0223] Assays of LFA-1, Mac-1, GPIIb/IIIa and .alpha.v.beta.3
involved coating the purified receptor on a 96-well microtiter
plate. The specific ligands for these receptors were labeled with
europium chelates. In the assays of LFA-1 and Mac-1, an ICAM-1 IgG
fusion protein containing the one to five immunoglobulin domains of
human ICAM-1 (D1D5) attached above the hinge region of an IgG1
molecule, was used. In the assays of GPIIb/IIIa and
.alpha.v.beta.3, europium-labeled fibrinogen and vitronectin,
respectively, was used. The purified receptors were allowed to
incubate in the wells with various concentrations of test compound,
in the presence of europium-labeled ligands. Following incubation,
the wells are emptied and washed. The amount of europium-labeled
ligand bound was determined by time-resolved fluorescence
measurement Assay of .alpha.4.beta.7 is similar to the adhesion
inhibition assay of VLA-4 described above, and uses the
.alpha.4.beta.7-expressing cell, RPMI-8886. A MAdCAM-1 IgG fusion
protein containing the one and two immunoglobulin domains of human
MAdCAM-1 and mucin-like repeat domain, is used as the corresponding
ligand for .alpha.4.beta.7.
[0224] Eu.sup.3+-VCAM-1 IgG binding to CHO/VLA-4 cells may be
determined as follows. 4B4 cells (CHO/VLA-4 cells) are distributed
into each well of a 96-well microtiter plate at
3.times.10.sup.4/well. The plate is incubated at 37.degree. C., 5%
CO.sub.2 for 48 hours and then washed twice with washing buffer,
then blot dried. 50 .mu.l of the inhibitor solution diluted with
assay buffer (2% DMSO final) is added to each well, followed by 50
.mu.l of Eu.sup.3+-VCAM-1 IgG diluted with assay buffer at 2 nM.
The plate is incubated at room temperature for at least 30 min.
Each well is then washed four times with washing buffer and blot
dried. 100 .mu.l of DELFIA Enhancement solution is added to each
well, followed by agitation of the plate at room temperature for 5
min. Fluorescence of each sample is then measured (e.g., DELFIA
Fluorometer 1234, Wallace, Inc., USA). In this assay, the washing
buffer comprises 25 mM HEPES (pH 7.5), 150 mM NaCl, 1 mM
CaCl.sub.2, 1 mM MgCl.sub.2, and 4 mM MnCl.sub.2; the assay buffer
comprises 25 mM HEPES (pH 7.5), 150 mM NaCl, 1 mM CaCl.sub.2, 1 mM
MgCl.sub.2, 4 mM MnCl.sub.2, 1% BSA, and 20 .mu.M DTPA.
In Vivo Assays
[0225] The VLA-4 inhibitors may be further characterized in in vivo
assays. One such assay examines the inhibition of eosinophil
infiltration into the bronchoalveolar lavage fluid in the mouse
(murine) model. In this assay, the animals are treated with
cyclophosphamide on day 0. On days 2 and 14, the animals are
immunized intraperitoneally with Ascaris suum extract. Seven days
later, the animals are treated with various doses of the VLA-4
inhibitor. Shortly after drug administration, the animals are
challenged with Ascaris suum extract by instillation into the
trachea. Bronchoalveolar lavage of the animal is performed by
instilling saline into the lung, 48 hours later. Total cell and
eosinophil counts in the lavage are determined.
[0226] In the murine model of Ascaris-induced bronchial
inflammation, one of the representative compounds (example number
32) inhibited eosinophil infiltration by 49% at an oral dosage of
30 mg/kg. By contrast, a representative prior art compound,
4-(N'-2-methylphenylurea)phenylacetyl-LDVP-OH, described in WO
97/03094, did not inhibit eosinophil infiltration (%
inhibition=-2%) at an oral dosage of 50 mg/kg.
[0227] Other representative compounds were also tested in mice. The
dosage, route of administration and inhibitory effect of
representative compounds (hereinafter, all tested compounds are
referenced by compound number provided in the Synthetic Examples)
are shown in Table 5. TABLE-US-00005 TABLE 5 % Inhibition of
Eosinophil Dosage/Regimen Route Infiltration Compound 79 30 mg/kg
b.i.d.sup.1 .times. 2 days i.v. 35.9 Compound 90 10 mg/kg
t.i.d.sup.2 .times. 2 days p.o. 18.1 Compound 90 30 mg/kg t.i.d.
.times. 2 days p.o. 39.1 Compound 90 50 mg/kg t.i.d. .times. 2 days
p.o. 45.9 Compound 90 30 mg/kg b.i.d. .times. 2 days i.v. 47.3
Compound 314 50 mg/kg t.i.d .times. 2 days p.o. 18.9 Compound 311
50 mg/kg t.i.d .times. 2 days s.c. 80.2 Compound 311 50 mg/kg t.i.d
.times. 2 days p.o. 42.5 Compound 311 50 mg/kg t.i.d .times. 2 days
s.c. 49.3 .sup.1two times a day (0 and 8 hrs) .sup.2three times a
day (0, 8 and 16 hrs)
[0228] The compounds of the present invention may also be further
characterized in other in vivo assays, such as the eosinophil
accumulation model tested in the rat. Fifty .mu.g of Compound 48/80
was injected into the pleural cavities of male Sprague Dawley rats.
After 24 hrs, each cavity was washed twice with Hank's Balanced
Salt Solution containing 0.2% EDTA. Total cell and eosinophil
counts were determined. Test compounds were given intravenously,
orally or subcutaneously, b.i.d. at 0 and 8 hours. The dosage,
route of administration and inhibitory effect for the test
compounds are shown in Table 6. TABLE-US-00006 TABLE 6 % Inhibition
of Eosinophil Dosage/Regimen Route Infiltration Compound 90 3 mg/kg
2 days i.v. 25.4 10 mg/kg 2 days i.v. 46.7 30 mg/kg 2 days i.v.
83.7 Compound 90 50 mg/kg 2 days p.o. 50.5 Compound 80 50 mg/kg 2
days s.c. 65.3 Compound 92 50 mg/kg 2 days s.c. 43.1 Compound 95 50
mg/kg 2 days s.c. 40.9
Mouse Bio-Assay Method
[0229] A compound was dissolved or suspended with an appropriate
solvent at 1 mg/mL. Female Balb/c mice (7-9 weeks old) were given
the compound orally. Blood samples were collected from the
postcaval vein of the anesthetized mice after fifteen minutes.
Serum was prepared and stored at -20.degree. C. Serum concentration
of the compound was determined from inhibitory activities of the
diluted serum by a direct binding assay using VLA-4-expressing
cells and VCAM-IgG fusion protein. Serum concentration determined
by this method correlated well with the concentration determined by
LC/MS/MS methodologies. The dosage, route of administration and
resulting inhibitory effect for the test compounds are shown in
Table 7. TABLE-US-00007 TABLE 7 Serum Minutes Post- Concentration
Dosage Administration (ng/ml) Compound 58 50 mg/kg 30 3614 Compound
68 10 mg/kg 15 261 Compound 78 10 mg/kg 15 368 Compound 79 10 mg/kg
15 618 Compound 80 10 mg/kg 15 693 Compound 90 10 mg/kg 15 3659
Compound 91 10 mg/kg 15 2523 Compound 92 10 mg/kg 15 2162 Compound
96 10 mg/kg 15 3514 Compound 97 10 mg/kg 15 1733 Compound 98 10
mg/kg 15 2796 Compound 102 10 mg/kg 15 503 Compound 124 10 mg/kg 15
841 Compound 134 10 mg/kg 15 224 Compound 146 10 mg/kg 15 527
Compound 156 10 mg/kg 15 285 Compound 158 10 mg/kg 15 301 Compound
166 10 mg/kg 15 360 Compound 179 10 mg/kg 15 428 Compound 309 10
mg/kg 15 669 Compound 311 10 mg/kg 15 467 Compound 314 50 mg/kg 30
2309 Compound 318 50 mg/kg 30 2105 Compound 319 50 mg/kg 15 603
Compound 323 50 mg/kg 15 1423
Pharmakokinetic Evaluations
[0230] Pharmacokinetic parameters of exemplary compounds, in mouse,
rat and monkey models, are shown in Tables 8, 9 and 10.
TABLE-US-00008 TABLE 8 MOUSE AUC.sup.1 MRT.sup.2 CL.sup.3 Dosage
(ng h/mL) (hr) (mL/min/kg) Compound 68 10 mg/kg (i.v.) 1595 (0-6
hr) 0.2 104.5 1595 (0-.infin.) 0.2 104.5 20 mg/kg (p.o.) 1307 (0-6
hr) 1.6 637.6 1751 (0-.infin.) 4.1 476.0 Compound 90 10 mg/kg
(i.v.) 8995 (0-6 hr) 0.5 18.53 9219 (0-.infin.) 0.7 18.08 10 mg/kg
(p.o.) 2540 (0-6 hr) 1.2 65.62 2950 (0-.infin.) 2.5 56.50 Compound
80 10 mg/kg (i.v.) 5259 (0-6 hr) 0.4 31.69 5389 (0-.infin.) 0.6
30.93 20 mg/kg (p.o.) 2190 (0-6 hr) 2.3 152.24 3615 (0-.infin.) 6.5
92.21 .sup.1total area under the plasma concentration (measured by
LC/MS/MS method) versus time curve .sup.2mean residence time
.sup.3apparent plasma clearance
[0231] TABLE-US-00009 TABLE 9 RAT AUC.sup.1 MRT.sup.2 CL.sup.3
Dosage (ng h/mL) (hr) (mL/min/kg) Compound 90 10 mg/kg (i.v.) 31915
(0-6 hr) 0.8 5.23 33488 (0-.infin.) 1.1 4.98 10 mg/kg (p.o.) 11454
(0-6 hr) 1.7 17.85 19968 (0-.infin.) 8.8 9.99 Compound 79 10 mg/kg
(i.v.) 5259 (0-6 hr) 0.4 31.69 5389 (0-.infin.) 0.6 30.93 10 mg/kg
(p.o.) 22119 (0-6 hr) 0.5 8.5 24867 (0-.infin.) 0.7 6.7 Compound 68
10 mg/kg (i.v.) 6345 (0-6 hr) 0.63 26.45 6405 (0-.infin.) 0.67
26.20 20 mg/kg (p.o.) 1867 (0-6 hr) 1.1 181.1 2086 (0-.infin.) 2.0
162.7 .sup.1total area under the plasma concentration (measured by
LC/MS/MS method) versus time curve .sup.2mean residence time
.sup.3apparent plasma clearance
[0232] Pharmacokinetic parameters and the time course of the serum
concentration of a single intravenous dosage (2 mg/kg) of a
representative compound and ATENOLOL
{4-[2'-hydroxy-3'-isopropylamino)proppyxy]phenylacetamide; Sigma
Chemical Co., code no. A-7655}are summarized in Tables 10 and 11
for the monkey model. TABLE-US-00010 TABLE 10 MONKEY AUC.sup.1
Cltot.sup.2 (ng h/mL) (mL/min/kg) Compound 195 8405 4.0 ATENOLOL
5021 6.7 .sup.1total area under the plasma concentration (measured
by the LC/MS/MS method) versus time curve .sup.2apparent plasma
clearance
[0233] TABLE-US-00011 TABLE 11 TIME 5 min 30 min 1 hr 2 hr 4 hr 8
hr Serum conc.sup.1 65447 3900 2225 772 194 28 (ng/ml) Compound 195
Serum conc.sup.1 4057 1189 771 479 348 137 (ng/ml) ATENOLOL
.sup.1measured by the LC/MS/MS method
Binding Assay of VCAM-1 to VLA-4 Expressing Cells
[0234] Preparation of VCAM IgG Fusion Protein
[0235] A VCAM IgG fusion protein containing the one to seven
immunoglobulin domains of VCAM-1 (D1D7) ligated to the hinge (H),
CH.sub.2 and CH.sub.3 regions of human IgG1 was used in the binding
assay.
Construction of a Stable Cell Line Expressing D1D7-VCAM IgG Fusion
Protein
[0236] An Epstein-Barr virus based, episomal plasmid containing a
D1D7-VCAM IgG fusion gene under transcriptional control of the CMV
promoter, was transfected into 293E human embryonic kidney cells.
Stably transfected cells were selected using 250 .mu.g/mL
hygromycin in DMEM with 10% fetal calf serum. The cells secreted
D1D7 VCAM IgG fusion protein into the medium cumulatively for up to
9 days.
[0237] Purification of D1D7 VCAM IgG Fusion Protein
[0238] The cells were cultured in DMEM with 10% fetal calf serum
for 2 days, then changed to CCM5 medium and cultured for a further
10 days. The medium was centrifuged, filtered and then incubated
overnight with Protein A Sepharose 4. The Protein A Sepharose was
washed extensively and the D1D7 VCAM IgG fusion protein bound was
eluted using 100 mM citric acid, pH 3.
[0239] Preparation of Europium Labeled-D1D7 VCAM IgG Fusion
Protein
[0240] The D1D7-VCAM IgG fusion protein, at 1 mg/mL, was dialyzed
against 50 mM NaHCO.sub.3, 0.9% NaCl, pH 8.5. The fusion protein
was added to one vial of europium-labeling reagent (DELFIA labeling
kit from Wallac, Gaithersberg, Md.; catalog no. 1244-302) and
incubated at room temperature in the dark overnight. The labeled
protein was purified using a Sepharose G10 column and assayed for
the europium content and protein concentration. The protein was
stored at minus 80.degree. C. until used.
[0241] Construction of Cell Line Expressing VLA-4 (CHO/VLA-4)
[0242] A CHO cell line stably transfected with the cDNA of .alpha.4
and .beta.1 was used in the binding assay. The gene for human
.alpha.4 was obtained from the American Type Culture Collection and
recloned between the XhoI and Xba sites of the mammalian expression
vector pCI-neo (Promega, Madison, Wis.). The .beta.1 gene was
amplified by PCR from human peripheral leukocyte cDNA and
engineered such that the start codon was placed in the context of a
consensus Kozak sequence. The gene was recloned into pCI-neo
downstream of the CMV promoter and chimeric intron.
[0243] CHO-K1 cells were stably co-transfected with plasmids
encoding the .alpha.4 and .beta.1 genes, and single cells
expressing high levels of VLA-4 were selected by fluorescence cell
sorting (FACS). The antibodies used in FACS analysis were:
anti-.alpha.4-PE conjugated (PharMingen, San Diego, Calif.) and
anti-.beta.1-FITC conjugated (Biosource, Camarillo, Calif.). A cell
line 4B4, which expresses 400,000 and 300,000 sites/cell of the
.alpha.4 and .beta.1 subunit, respectively, was used in the binding
assay. The subunit numbers were determined by FACS analysis, using
Quantum Simply Cellular microbeads (Flow Cytometry Standards
Corporation, Puerto Rico) as standards. The cells were maintained
in F12 medium, containing 10% fetal bovine serum, 10 mM HEPES, pH
7.5, 0.5 mg/mL G418, using a 1:48 passage/week.
Binding Assay
[0244] The CHO/VLA-4 cells were seeded in a 96-well microtiter
plate at 30,000 cells/well and incubated at 37.degree. C., 5%
CO.sub.2 for 48 hours until confluent. On the day of assay, the
wells were emptied and washed twice with 350 .mu.l of a washing
buffer containing 25 mM HEPES, pH 7.5, 150 mM NaCl, 1 mM
MgCl.sub.2, 1 mM CaCl.sub.2, 2 mM MnCl.sub.2. The plate was then
drained and blotted dry on paper towels to remove buffer.
[0245] The test compound was serially diluted in assay buffer
(washing buffer together with 0.1% bovine serum albumin, 20 .mu.M
DTPA and 1% dimethylsulfoxide), in the presence of 2 nM of
europium-labeled D1D7-VCAM IgG fusion protein. Final concentrations
used ranged from 0.1 nM-10 .mu.M. 50 .mu.l aliquot of the test
compound mixture was added to duplicate wells in the plate. Control
wells for total binding received no test compound. Non-specific
binding wells contained an anti-4 monoclonal antibody (L25.3,
Becton Dickinson, Bedford, Mass.).
[0246] The cells were allowed to incubate with the test compound
mixture, in the presence of europium-labeled D1D7-VCAM IgG fusion
protein at room temperature for at least 30 minutes. The cells were
then washed three times with 350 .mu.l of washing buffer, using a
Skatron plate washer and blot dry. An 100 .mu.l aliquot of DELFIA
Enhancement solution was added to each well, followed by gentle
agitation at room temperature for 10 minutes. The amount of
europium-labeled VCAM-IgG fusion protein bound was determined by
time-resolved fluorescence measurement (Model: Victor.TM., Wallac
Inc., Gaithersberg, Md.).
[0247] Percent binding was calculated as:
[(F.sub.T-F.sub.NS)-(F.sub.1-F.sub.NS)]/(F.sub.T-F.sub.NS).times.100
wherein F.sub.T and F.sub.NS is the fluorescence signal of the
europium labeled D1D7-VCAM IgG fusion protein bound to cells, in
the absence of test compound and containing an anti-.alpha.4
monoclonal antibody, respectively. F.sub.1 is the fluorescence in
wells containing a test compound. The IC.sub.50 (concentration of
the inhibitor to inhibit 50% binding of VACM to CHO/VLA-4 cell) was
determined by a curve fitting routine, PRIZM (GraphPad Software,
Inc., San Diego, Calif.).
Adhesion of VLA-4 Expressing Cell to VCAM-1
[0248] This secondary functional assay was used to determine the
potency of a test compound in inhibiting VLA-4 mediated cell
adhesion.
[0249] Preparation of VCAM Coated Plate
[0250] A 50 .mu.l aliquot of the D1D7-VCAM IgG fusion protein (1.5
.mu.g/mL in phosphate buffered saline, PBS) was added to each well
of a 96-well Costar flat bottom plate (Costar, Franklin Lakes,
N.J., catalog no. 2580). The plate was then incubated overnight at
4.degree. C. On the day of assay, the wells were emptied and washed
twice with 350 .mu.l of PBS. The plate was then blocked with 100
.mu.l of 1% bovine serum albumin (BSA, Sigma, cat# A9418) in PBS at
room temperature for at least a hour.
[0251] Cell Preparation
[0252] Jurkat cell (clone E6-1) was obtained from American Type
Cultured Collection and was maintained in RPMI medium, 10 mM HEPES,
pH 7.5, 1 mM sodium pyruvate, 10% FCS, using a 1:64 passage/week.
Just prior to running the assay, Jurkat cells were labeled with 5
.mu.M of calcein-AM (Molecular Probe, Eugene, Oreg., catalog no.
C1430) in RPMI medium, at room temperature for 30 min in the dark.
Following labeling, cells were washed twice with RPMI medium and
resuspended at 1.times.10.sup.6 cells/mL.
[0253] Cell Adhesion Assay
[0254] Immediately before the assay, the BSA solution was emptied
from the VCAM-coated plate. The plate was then washed twice with
RPMI medium. A 100 .mu.l aliquot of the labeled Jurkat cells was
added to each well, followed by the addition of 50 .mu.l of the
inhibitor solutions. Final inhibitor concentrations range from 1 nM
to 10 .mu.M and each concentration was tested in triplicates. The
inhibitor and cells were allowed to incubate at room temp for 1 hr
in the dark. Following the incubation, the plate was immersed
gently into a container filled with PBS, then inverted face down
under PBS. The wells were drained and blotted dry on a layer of
paper towel. A 50 .mu.l aliquot of 0.1% Triton X-100 was added to
each well. The plate was incubated in the dark for 10 min. Adhesion
of Jurkat cell was quantitated in a Millipore Cytofluor 2300 System
plate reader set at 485 nM excitation and 530 nM emission. The
IC.sub.50 (concentration of the inhibitor to inhibit 50% Jurkat
cell adhesion) was determined by a curve fitting routine, PRIZM
(GraphPad Software, Inc., San Diego, Calif.).
Methods of Synthesis
[0255] Compounds of the present invention may be prepared by
standard chemical synthesis methods, as well as by methods of
combinatorial chemistry, such as that described in Published PCT
application, WO 95/30642.
SYNTHETIC EXAMPLES
[0256] General methods of synthesis are illustrated by the
following examples. The specific embodiments are presented by way
of illustration only, and are not intended to limit the invention.
Modifications and variations in any given material or process step
will be readily apparent to one of skill in the art. Unless
otherwise indicated, the solid-phase support used in certain
examples is Tentagel.TM.-S-PHB resin. This resin has a para-hydroxy
benzyl linker which can be cleaved by the use of 90%
trifluoroacetic acid in dichloromethane. The loading for this resin
varies between 0.27 and 0.30 mmol/g and is not double loaded.
Example 1
[0257] ##STR1616##
[0258] A three-necked 500 mL round-bottomed flask was charged with
200 mL of THF and NaH (1.5 g, 62.9 mmol). A solution of
1-vinyl-2-pyrrolidinone (6.9 g, 62.9 mmol) and methyl
3-iodobenzoate (15.0 g, 57.3 mmol) in THF (100 mL) was added
dropwise to the flask over 15 min. After the addition was complete
the reaction mixture was heated to reflux for 1 hr. The reaction
vessel was allowed to cool to room temp and then 6 N HCl (100 mL)
was carefully added. The reaction was concentrated in vacuo to
remove the THF and then an additional aliquot of 6 N HCl (100 mL)
was added and the reaction was refluxed for 14 hr. The reaction was
quenched by the addition of NaHCO.sub.3 until pH 9 and then the
mixture was extracted 3.times. with EtOAc. The combined organics
were dried over MgSO.sub.4 and concentrated in vacuo to afford a
yellow oil.
[0259] This oil was then placed in MeOH (100 mL) and cooled to
minus 78.degree. C. NaBH.sub.4 (3.5 g, 96.5) was then added
portionwise and the reaction was allowed to warm to room temp over
2 hr. The reaction was quenched by the addition of 6 N HCl until
acidic and then made basic by the addition of 40% aqueous NaOH, The
solution was extracted 3.times. with CH.sub.2Cl.sub.2, the combined
organics were dried over MgSO.sub.4, and then concentrated in vacuo
to afford 11.4 g as a yellow oil.
[0260] The above amine was then Boc-protected by placing the amine
in 50% dioxane:H.sub.2O (100 mL) and adding K.sub.2CO.sub.3 until
basic. To this solution was added Boc-anhydride (9.1 g, 41 mmol)
and then allowed to stir for 14 hr at room temp. The reaction was
quenched by the addition of 1 N HCl until acidic. The solution was
extracted 3.times. with EtOAc, dried over MgSO.sub.4 and
concentrated in vacuo to afford a yellow viscous oil. The oil was
chromatographed (25% EtOAc:hexanes) to afford 7.0 g A.
##STR1617##
[0261] Hydrochloric acid (gas) was bubbled through 15.8 g (73.5
mmol) 2-bromophenylacetic acid in 100 mL of methanol for 10 min.
The resulting solution was partitioned between 100 mL water and 100
mL CH.sub.2Cl. The organic layer was dried over MgSO.sub.4 and the
solvent was removed under reduced pressure to give 16.8 g (73.5
mmol) of methyl-2-bromophenylacetate which was combined with 9.0 g
(80.8 mmol) of 1-vinyl-2-pyrrolidinone, and 100 mL of dry THF under
argon in a 250 mL round-bottomed flask.
[0262] To this flask was added 3.5 g (147 (mmol) sodium hydride
(95%) and the solution was stirred for 10 min at room temp. A
reflux condenser was added and the mixture was heated to reflux for
1 hr. The solution was cooled to room temp and the solvent was
removed under reduced pressure. A solution of 30 mL aqueous
hydrochloric acid and 50 mL water was added to the resultant
mixture and was heated to reflux with no condenser until the
solution temperature reached 96.degree. C. at which time a
condenser was added and the solution was allowed to reflux for 16
hr. The solution was cooled to room temp, made basic with 150 mL of
an aqueous solution of 40% sodium hydroxide, extracted with
3.times.125 mL CH.sub.2Cl.sub.2, and the combined organic layers
were washed with a saturated aqueous solution of sodium chloride,
dried over magnesium sulfate, and the solvent was removed under
educed pressure to give 15.0 g (63.0 mmol, 86%) of
2-(2-bromobenzyl)-1-pyrolline.
[0263] To a solution of 15 g (63 mmol) of
2-(2-bromobenzyl)-1-pyrolline in a solution of 80:20
methanol:aqueous acetic acid cooled to minus 78.degree. C. was
added, in portions over a 15 min period, 5.3 g (140.0 mmol) sodium
borohydride. The mixture was allowed to stir for 1 hr warming to
room temp at which time the solvent was removed under reduced
pressure, 150 mL of water was added and the solution was made basic
with an aqueous solution of sodium hydroxide which was extracted
10.times.100 mL CH.sub.2Cl.sub.2 which resulted in emulsions. The
combined organic layers were washed with a saturated aqueous
solution of sodium bicarbonate, dried over magnesium sulfate and
the solvent was removed in vacuo to give 14.6 g (60.8 mmol, 97%) of
the benzyl proline.
[0264] To a solution of 14.6 g (60.8 mmol) of the benzyl proline in
a solution of 70 mL of saturated aqueous sodium bicarbonate and 70
mL dioxane was added 15.1 g (67.0 mmol) of di-t-butyl-dicarbonate
and the mixture was stirred for 16 hr at room temp. The solution
was then partitioned between a 200 mL aqueous solution of
hydrochloric acid and 200 mL ethyl acetate. The ethyl acetate layer
was washed with 200 mL of a saturated aqueous solution of sodium
chloride, dried over MgSO.sub.4 and the solvent was removed under
reduced pressure to give a residue which was purified by flash
column chromatography (20% 100% ethyl acetate/hexane) to give 11.5
g (33.8 mol, 56%) pure A'. ##STR1618##
[0265] A (1.88 g, 5.0 mmol) was placed in a 100 mL round-bottomed
flask and dissolved in DMF (50 mL). To this solution was added
Pd(OAc).sub.2 (23 mg, 0.3 mmol), P(o-Tol).sub.3 (12 mg, 0.3 mmol),
methyl acrylate (0.47 g, 5.5 mmol), and NaOAc (0.5 g, 5.5 mmol).
This mixture was then heated to 80.degree. C. for 14 hr. The
reaction mixture was then cooled to room temp and 1 N HCl (100 mL)
was added. The solution was then extracted 3.times. with EtOAc,
dried over MgSO.sub.4, and then concentrated in vacuo to afford a
brown oil. This oil was chromatographed with 25% EtOAc:hexanes to
afford 1.32 g of the alkene ester as a colorless viscous oil.
[0266] The alkene ester (1.32 g, 4.1 mmol) was then subjected to
hydrogenation. The alkene was placed in a Parr hydrogenation
bottle, EtOAc (10 mL) and 10% Pd/C (100 mg) was added under inert
atmosphere. The bottle was then pressurized with hydrogen at 45 psi
and shaken for 4 hr at room temp. The solution was then filtered
through celite and concentrated in vacuo to afford 1.29 g of the
alkane ester.
[0267] The alkane ester (1.29 g, 4.0 mmol) was dissolved in THF (30
mL), MeOH (20 mL), and water (10 mL) and saponified with LiOH (200
mg, 8.0 mmol). The reaction was stirred at room temp for 3 hr and
then poured into 1 N HCl (50 mL). This solution was then extracted
3.times. with EtOAc, dried over MgSO.sub.4, and then concentrated
in vacuo to afford 1.02 g of the alkane acid as a yellow solid.
[0268] The alkane acid (1.02 g, 3.3 mmol) was then deprotected by
the addition of a 25% TFA/CH.sub.2Cl.sub.2 solution and stirred for
2 hr at room temp. The resulting mixture was then concentrated in
vacuo and immediately protected by dissolving the deprotected acid
in 50% dioxane/water, adding K.sub.2CO.sub.3 (1.2 g), and Fmoc-Cl
(1.08 g, 4.0 mmol). This mixture was stirred at room temp for 14 hr
and then poured in 1 N HCl (100 mL). The solution was then
extracted 3.times. with EtOAc, dried over MgSO.sub.4, and then
concentrated in vacuo to afford 495 mg 1 as a white crystalline
solid. ##STR1619##
[0269] The dried resin (500 mg, 0.14 mmol) was placed into a small
shaker vessel. The vessel was then charged with 9 mL of DMF (2 mg,
0.42 mmol), DIC (102 mg, 0.84 mmol), and DMAP (17 mg, 0.14 mmol).
The vessel was subsequently shaken for 16 hr at room temp. The
contents were drained and the resin was washed 3.times. with DMF,
MeOH, and CH.sub.2Cl.sub.2. The Fmoc group was then removed by the
addition of 10 mL of 50% piperidine/DMF to the shaker vessel and
shaking for 2 hr at room temp. The resulting amine resin was washed
3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2.
[0270] To the above resin was added 9 mL of DMF,
4-[N'-(o-Tolylurea)]-phenylacetic acid (132 mg, 0.42 mmol), PyBroP
(196 mg, 0.42 mmol), and DIEA (107 mg, 0.84 mmol). The contents
were shaken for 14 hr at room temp and then drained and washed
3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2. The compound was
then cleaved from the resin and the filtrate was collected and then
concentrated in vacuo. The resulting oil was triturated by taking
up the oil in MeOH and slowly adding in Et.sub.2O until a
precipitate formed. This precipitate was collected and dried in
vacuo to afford 67 mg 1' as a white crystalline solid.
Example 2
[0271] ##STR1620##
[0272] A (4.2 g, 11.3 mmol) was placed in a 100 mL round-bottomed
flask and dissolved in NEt.sub.3 (50 mL). To this solution was
added Pd(PPh.sub.3).sub.2Cl.sub.2 (0.16 g, 0.23 mmol), CuI (21 mg,
0.12 mmol), and trimethylsilylacetylene (1.38 g, 13.5 mmol). This
mixture was stirred at room temp for 14 hr. The reaction mixture
was quenched by the addition of 1 N HCl (100 mL). The solution was
then extracted 3.times. with EtOAc, dried over MgSO.sub.4, and then
concentrated in vacuo to afford a yellow oil. This oil was
chromatographed with 15% EtOAc:hexanes to afford 3.8 g 2 as a
colorless viscous oil. ##STR1621##
[0273] A solution of dicyclohexylborane was generated by the
addition of borane-THF (12.0 mL, 12 mmol), at 0.degree. C. to a
solution of cyclohexene (2.3 mL) in 6 mL of anhydrous THF. This
solution was stirred for an additional 1 hr at 0.degree. C. The
acetylene (2) (2.0 g, 5.84 mmol) was then added dropwise over 15
min at 0.degree. C. and then allowed to warm to room temp over 1
hr. The reaction mixture was then diluted with MeOH (20 mL) and
then recooled to 0.degree. C. A solution of 2 N NaOH (6 mL) and 30%
H.sub.2O.sub.2 (3.5 mL) was then added dropwise. The reaction
mixture was then stirred at 0.degree. C. for 1 hr and then warmed
to 40.degree. C. for 2.5 hr. The mixture was then cooled to room
temp and an additional 6 mL of 2 N NaOH was added. The organics
were removed in vacuo and the remaining aqueous solution was
extracted 3.times.Et.sub.2O and the organics were discarded. The
aqueous extracts were then acidified with 1 N HCl and extracted
with EtOAc dried over MgSO.sub.4, and then concentrated in vacuo to
afford 1.7 g of the phenylacetic acid as a tan crystalline
solid.
[0274] The acid (1.7 g, 5.6 mmol) was then deprotected by the
addition of a 25% TFA/CH.sub.2Cl.sub.2 solution and stirred for 2
hr at room temp. The resulting mixture was then concentrated in
vacuo and immediately protected by dissolving the deprotected acid
in 50% dioxane/water, adding K.sub.2CO.sub.3 (15 g), and Fmoc-Cl
(1.4 g, 5.5 mmol). This mixture was stirred at room temp for 14 hr
and then poured in 1 N HCl (100 mL). The solution was then
extracted 3.times. with EtOAc, dried over MgSO.sub.4, and then
concentrated in vacuo to afford 1.7 g 3 as a white crystalline
solid. ##STR1622##
[0275] The dried resin (500 mg, 0.14 mmol) was placed into a small
shaker vessel. The vessel was then charged with 9 mL of DMF, 3 (180
mg, 0.42 mmol), DIC (102 mg, 0.84 mmol), and DMAP (17 mg, 0.14
mmol). The vessel was subsequently shaken for 16 hr at room temp.
The contents were drained and the resin was washed 3.times. with
DMF, MeOH, and CH.sub.2Cl.sub.2. The Fmoc group was then removed by
the addition of 10 mL of 50% piperidine/DMF to the shaker vessel
and shaking for 2 hr at room temp. The resulting amine resin was
washed 3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2.
[0276] To the above resin was added 9 mL of DMF,
4-[N'-(o-Tolylurea)]-phenylacetic acid (132 mg, 0.42 mmol), PyBroP
(196 mg, 0.42 mmol), and DIEA (107 mg, 0.84 mmol). The contents
were shaken for 14 hr at room temp and then drained and washed
3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2. The compound was
then cleaved from the resin and the filtrate was collected and then
concentrated in vacuo. The resulting oil was triturated by taking
up the oil in MeOH and slowly adding in Et.sub.2O until a
precipitate formed. This precipitate was collected and dried in
vacuo to afford 52 mg 3' as a white crystalline material.
Example 3
[0277] ##STR1623##
[0278] The acetylene (2) was deprotected by placing 2 (0.75 g, 2.1
mmol) in MeOH (25 mL) and adding to this solution KOH (1.4 g). The
resulting solution was stirred at room temp for 1 hr. The reaction
mixture was then concentrated in vacuo and acidified with 1 N HCl.
The resulting aqueous solution was extracted 3.times.EtOAc, the
combined organics were dried over MgSO.sub.4, and then concentrated
in vacuo to afford 0.56 g of the deprotected acetylene as a brown
oil.
[0279] The deprotected acetylene (0.56 g, 2.0 mmol) was then placed
into THF (50 mL) and cooled to -78.degree. C. LiHMDS (1 M soln, 4.7
mL) was then added dropwise and the reaction was stirred for 30
min. CO.sub.2 gas was then bubbled through the reaction mixture for
15 min and the reaction was then poured onto CO.sub.2 solid. The
reaction was quenched by the addition of 1 N HCl (100 mL) and the
aqueous solution was extracted 3.times.EtOAc, the combined organics
were dried over MgSO.sub.4, and then concentrated in vacuo to
afford the propionic acid (0.71 g) as a white solid.
[0280] The alkane acid (0.71 g) was then deprotected by the
addition of a 25% TFA/CH.sub.2Cl.sub.2 solution and stirred for 2
hr at room temp. The resulting mixture was then concentrated in
vacuo and immediately protected by dissolving the deprotected acid
in 50% dioxane/water, adding K.sub.2CO.sub.3 (15 g), and Fmoc-Cl
(1.29 g, 4.9 mmol). This mixture was stirred at room temp for 14 hr
and then poured in 1 N HCl (100 mL). The solution was then
extracted 3.times. with EtOAc, dried over MgSO.sub.4, and then
concentrated in vacuo to afford 4 as a brown oil. The oil was then
chromatographed with 5% MeOH/dichloromethane to afford 110 mg of
the desired compound. ##STR1624##
[0281] The dried resin (500 mg, 0.14 mmol) was placed into a small
shaker vessel. The vessel was then charged with 9 mL of DMF, 4 (184
mg, 0.42 mmol), DIC (102 mg, 0.84 mmol), and DMAP (17 mg, 0.14
mmol). The vessel was subsequently shaken for 16 hr at room temp.
The contents were drained and the resin was washed 3.times. with
DMF, MeOH, and CH.sub.2Cl.sub.2. The Fmoc group was then removed by
the addition of 10 mL of 50% piperidine/DMF to the shaker vessel
and shaking for 2 hr at room temp. The resulting amine resin was
washed 3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2.
[0282] To the above resin was added 9 mL of DMF,
4-[N'-(o-Tolylurea)]-phenylacetic acid (132 mg, 0.42 mmol), PyBroP
(196 mg, 0.42 mmol), and DIEA (107 mg, 0.82 mmol). The contents
were shaken for 14 hr at room temp and then drained and washed
3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2. The compound was
then cleaved from the resin and the filtrate was collected and then
concentrated in vacuo. The resulting oil was triturated by taking
up the oil in MeOH and slowly adding in Et.sub.2O until a
precipitate formed. This precipitate was collected and dried in
vacuo to afford 27 mg 4' as a white crystalline material.
Example 4
[0283] ##STR1625##
[0284] The iodide (A) (0.5 g, 1.3 mmol) was placed into THF (20 mL)
and cooled to minus 78.degree. C. Butyllithium (2.21 mL, 1.6M soln)
was added dropwise and then the cooling bath was removed and
gaseous CO.sub.2 was bubbled through for 10 min. The reaction
mixture was poured onto dry ice and then 1 M HCl (100 mL) was
added. The mixture was extracted 3.times.EtOAc, the combined
organics were dried over MgSO.sub.4, and then concentrated in vacuo
to afford 0.32 g of the benzoic acid as a white crystalline
solid.
[0285] The benzoic acid (0.32 g, 1.68 mmol) was then deprotected by
the addition of a 25% TFA/CH.sub.2Cl.sub.2 solution and stirred for
2 hr at room temp. The resulting mixture was then concentrated in
vacuo and immediately protected by dissolving the deprotected acid
in 50% dioxane/water, adding K.sub.2CO.sub.3 (15 g), and Fmoc-Cl
(0.44 g, 1.67 mmol). This mixture was stirred at room temp for 14
hr and then poured in 1 N HCl (100 mL). The solution was then
extracted 3.times. with EtOAc, dried over MgSO.sub.4, and then
concentrated in vacuo to afford 0.38 g 5 as a white crystalline
solid. ##STR1626##
[0286] The dried resin (500 mg, 0.14 mmol) was placed into a small
shaker vessel. The vessel was then charged with 9 mL of DMF, 5 (173
mg, 0.42 mmol), DIC (102 mg, 0.84 mmol), and DMAP (17 mg, 0.14
mmol). The vessel was subsequently shaken for 16 hr at room temp.
The contents were drained and the resin was washed 3.times. with
DMF, MeOH, and CH.sub.2Cl.sub.2. The Fmoc group was then removed by
the addition of 10 mL of 50% piperidine/DMF to the shaker vessel
and shaking for 2 hr at room temp. The resulting amine resin was
washed 3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2.
[0287] To the above resin was added 9 mL of DMF,
4-[N'-(o-Tolylurea)]-phenylacetic acid (132 mg, 0.42 mmol), PyBroP
(196 mg, 0.42 mmol), and DIEA (107 mg, 0.84 mmol). The contents
were shaken for 14 hr at room temp and then drained and washed
3.times. with DMF, MeOH, and CH.sub.2Cl.sub.2. The compound was
then cleaved from the resin and the filtrate was collected and then
concentrated in vacuo. The resulting oil was triturated by taking
up the oil in MeOH and slowly adding in Et.sub.2O until a
precipitate formed. This precipitate was collected and dried in
vacuo to afford 51 mg 5' as a white crystalline material.
Example 5
(E)-4-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]eth-
enyl]benzoic acid
[0288] ##STR1627##
[0289] To a cold (minus 78.degree. C.), stirred solution of
triethyl 4-phosphonomethylbenzoate (904 mg, 3.01 mmol) in THF (20
mL) was added LiHMDS (1.0 M in THF, 3 mL, 3.00 mmol) and the
stirring was continued for 1 hr at the same temp. N-Boc prolinal
(500 mg, 2.51 mmol) in THF (10 mL) was added to this mixture and
the mixture was allowed to warm to room temp for over 1 hr. After
being stirred for 2 hr, the mixture was quenched by water and
extracted with EtOAc. The extract was washed with brine (200 mL),
dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with n-hexane-EtOAc (8:1, v/v) as
eluent to give 713 mg (82%) ethyl
(E)-4-[2-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]ethenyl]benzoate
as a colorless crystalline solid. mp 68-70.degree. C.; IR (KBr)
1710, 1697, 1681 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta.1.39
(12H, series of m), 1.77-1.93 (3H, m), 2.11 (1H, m), 3.47 (2H, m),
4.34-4.54 (total 3H, m), 6.22 (1H, m), 6.43 (1H, d, J=14.2 Hz),
7.39 (2H, J=8.3 Hz), 7.97 (2H, d, J=8.3 Hz); MS (FAB) m/z 346
(M.sup.++1); Anal. Calcd for C.sub.20H.sub.27NO.sub.4: C, 69.54; H,
7.88; N, 4.05. Found: C, 69.52; H, 8.08; N, 4.07.
[0290] To a stirred solution of ethyl
(E)-4-[2-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]ethenyl]benzoate
(700 mg, 2.03 mmol) in CH.sub.2Cl.sub.2 (3 mL) was added TFA (3 mL)
and the resulting mixture was stirred for 3 hr. The mixture was
concentrated and the residue was made basic by the addition of sat.
NaHCO.sub.3. The mixture was extracted with CHCl.sub.3 (2.times.100
mL). The combined extracts were dried over Na.sub.2CO.sub.3 and
concentrated in vacuo to give 434 mg (87%) ethyl
(E)-4-[2-(2-pyrrolidinyl)ethenyl]benzoate as a brown oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.39 (3H, t, J=7.3 Hz), 1.52-2.06
(4H, series of m), 2.93-2.99 (1H, m), 3.07-3.13 (1H, m), 3.74 (1H,
q, J=7.3 Hz), 4.37 (2H, q, J=7.3 Hz), 6.34 (1H, dd, J=15.6, 7.3
Hz), 6.54 (1H, d, J=15.6 Hz), 7.41 (2H, d, J=8.3 Hz), 7.97 (2H, d,
J=8.3 Hz).
[0291] A mixture of ethyl (E)-4-[2-(2-pyrrolidinyl)ethenyl]benzoate
(434 mg, 1.77 mmol), pentafluorophenyl
4-[N'-(2-methylphenyl)ureido]phenylacetate (797 mg, 1.77 mmol),
Et.sub.3N (0.37 mL, 2.66 mmol) in DMF (15 mL) was stirred for 15
hr. The mixture was diluted with EtOAc (300 mL). The solution was
washed with brine (2.times.200 mL), dried over MgSO.sub.4, and
evaporated off in vacuo. The residue was chromatographed on
silica-gel with CHCl.sub.3-EtOAc (4:1) as eluent to give 906 mg
(q.y.) ethyl
(E)-4-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]et-
henyl]benzoate as a brown oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.39 (3H, t, J=7.3 Hz), 1.83-2.20 (4H, series of m), 2.24 (3H, d,
J=4.9 Hz), 3.63 (4H, m), 4.36 (2H, q, J=7.3 Hz), 4.62 and 4.84
(total 1H, m), 6.18-6.47 (2H, m), 7.03-8.02 (14H, series of m).
[0292] A stirred mixture of ethyl
(E)-4-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]et-
henyl]benzoate (906 mg, 1.77 mmol) in 0.25 N NaOH (14 mL) and THF
(14 mL) was heated under reflux for 3 days. The mixture was poured
into ice-1N HCl (200 mL) and the precipitate was collected with
suction. The solid was recrystallized from CHCl.sub.3-MeOH-n-hexane
to give 453 mg (53%) 6 as a light yellow crystalline powder. mp
165-168.degree. C.; IR (KBr) 3282, 2974, 2663, 2537, 1700, 1685
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.74-2.12 (4H, m),
2.24 (3H, d, J=4.9 Hz), 3.35-3.66 (4H, m), 4.67-4.74 (1H, m),
6.25-6.41 (1H, m), 6.53 (1H, s), 6.93 (1H, t, J=7.3 Hz), 7.08-7.92
(12H, series of m), 9.00 (1H, m), 12.87 (1H, br s); MS (FAB) m/z
484 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.29N.sub.3O.sub.4.0.5H.sub.2O: C, 70.71; H, 6.14; N,
8.39. Found: C, 70.46; H, 6.07; N, 8.39.
Example 6
4-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]ethyl]b-
enzoic acid
[0293] ##STR1628##
[0294] A mixture of
(E)-4-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]et-
henyl]benzoate (200 mg, 0.414 mmol) and 5% Pd/C (200 mg) in MeOH
(20 mL) was hydrogenated at 1 atm for 1 hr with vigorously
stirring. The mixture was filtered and the filtrate was
concentrated. The residue was chromatographed on silica-gel with
CHCl.sub.3-MeOH (4:1) as eluent to give 201 mg (q.y.) 7 as a
colorless crystalline powder. mp 180-190.degree. C.; IR (KBr) 3345,
3124, 3060, 3027, 2960, 2927, 2875, 1706, 1672 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.04-3.96 (total 16H, series of
m), 6.91-7.41 (9H, m), 7.79-7.90 (3H, m), 8.23 (1H, br s), 9.31
(1H, br s); MS (FAB) m/z 486 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.4.2.25H.sub.2O: C, 66.21; H, 6.80; N,
7.99. Found: C, 65.97; H, 6.20; N, 7.72.
Example 7
(S)-4-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]pyrrolidinyl]methox-
y]benzoic acid
[0295] ##STR1629##
(S)-4-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]pyrrolidinyl]methox-
y]benzoic acid
[0296] ##STR1630##
(S)-4-[2-[1-[4-[N'-(2-chlorophenyl)ureido]phenylacetyl]pyrrolidinyl]methox-
y]benzoic acid
[0297] ##STR1631##
[0298] 8, 9 and 10 To a stirred mixture of Boc-prolinol (3.00 g,
14.9 mmol), ethyl p-hydroxybenzoate (2.40 g, 14.5 mmol), and
triphenylphosphine (3.91 g, 14.9 mol) in THF (80 mL) was added
dropwise diethyl azodicarboxylate (2.86 g, 16.4 mmol) at room temp.
After the addition was completed, the resulting mixture was heated
under reflux for 2 hr. After cooling, the mixture was concentrated
in vacuo. The residue was dissolved in EtOAc and washed
successively with 1 N NaOH, water, brine. The EtOAc layer was dried
over MgSO.sub.4 and evaporated in vacuo. The residue was purified
by column chromatography on silica gel with EtOAc-n-hexane (1:4,
v/v) as eluent to give 4.88 g (93%) ethyl
(S)-4-(1-(tert-butoxycarbonyl-2-pyrrolidinyl)methoxybenzoate as an
oil.
[0299] To the above ethyl
(S)-4-(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxybenzoate was
added MeOH (100 mL) and 1 N NaOH (50 mL). The mixture was stirred
for 15 hr at room temp. After removal of MeOH under a reduced
pressure, water (50 mL) was added to the residual solution. The
aqueous solution was washed with Et.sub.2O (.times.2) and then
acidified by the addition of 1 N HCl. The mixture was extracted
with EtOAc., washed with water, brine, dried over MgSO.sub.4 and
evaporated in vacuo to afford 4.26 g (95%)
(S)-4-(1-tert-butoxycarbonyl-2-pyrrolidinyl) methoxybenzoic acid as
a crystalline solid.
[0300] To the above
(S)-4-(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxybenzoic acid was
added CH.sub.2Cl.sub.2 (10 mL) and TFA (10 mL). The mixture was
stirred at room temp for 1 hr Et.sub.2O was added to the mixture
and resulting solid was collected. The solid was dissolved in water
(100 mL), dioxane (50 mL) and NaHCO.sub.3 (4.4 g). Fmoc-Cl (3.34 g,
12.9 mmol) was added to the solution, and the resulting mixture was
stirred for 20 hr at room temp. The mixture was washed with
Et.sub.2O (.times.2) and aqueous layer was separated. The layer was
acidified by the addition of 1 N HCl. The mixture was extracted
with EtOAc. The extract was washed with water, brine, dried over
MgSO.sub.4 and evaporated in vacuo to afford 5.36 g (91%)
(S)-4-(1-Fmoc-(2-pyrrolidinyl)methoxybenzoic acid as a viscous oil,
which was crystallized on standing.
[0301] Wang resin (0.71 mmol/g, 400 mg) was suspended in a solution
of (S)-4-(1-Fmoc-2-pyrrolidinyl) methoxybenzoic acid (500 mg, 1.13
mol), DMAP (35 mg, 0.29 mmol), HOBt (40 mg, 0.30 mmol) and DIC
(0.45 mL, 2.9 mmol) in a mixture of DMF (3 mL) and CH.sub.2Cl.sub.2
(7 mL). The mixture was shaken for 20 hr and drained. The resin was
washed with DMF (.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2
(.times.3) and dried under a reduced pressure to give 522 mg of
resin, which was used to prepare 8, 9 and 10.
[0302] 8 To the above resin (115 mg) was added a solution of
piperidine-DMF (50% v/v, 4 mL) and the mixture was shaken for 1 hr.
The resin was washed with DMF (.times.3), MeOH (.times.3),
CH.sub.2Cl.sub.2 (.times.3). To the resin was added DMF (4 mL),
CH.sub.2Cl.sub.2 (2 mL), 4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (70 mg, 0.25 mmol), PyBrop (115 mg, 0.25 mmol) and DIEA (0.13
mL, 0.75 mmol). The mixture was shaken for 21 hr and drained. The
resin was washed with DMF (.times.3), MeOH (.times.3),
CH.sub.2Cl.sub.2 (.times.3). To the resin was added a solution of
TFA in CH.sub.2Cl.sub.2 (50% v/v, 4 mL) and the mixture was shaken
for 2 hr. The mixture was filtered and the filtrate was
concentrated in vacuo. The residue was purified by Sep-Pak column.
After removal of the solvent, Et.sub.2O was added to the residue
and resulting solid was collected to afford 25 mg 8 as a pale
yellow crystalline material. MS (FAB) m/z 488 (M.sup.++1)
[0303] 9 To the above resin (60 mg) was added a solution of
piperidine in DMF (50% v/v, 3 mL) and the mixture was shaken for 2
hr. The resin was washed with DMF (.times.3), MeOH (.times.3),
CH.sub.2Cl.sub.2 (.times.3). To the resin was added DMF (2 mL),
CH.sub.2Cl.sub.2 (1 mL) 3-methoxy-4-(N'-phenylureido)phenylacetic
acid (40 mg, 0.13 mmol), PyBrop (60 mg, 0.13 mmol) and DIEA (0.060
mL, 0.34 mmol). The mixture was shaken for 40 hr and drained. The
resin was washed with DMF (.times.3), MeOH (.times.3),
CH.sub.2Cl.sub.2 (.times.3). To the resin was added a solution of
TFA in CH.sub.2Cl.sub.2 (30% v/v, 3 mL) and the mixture was shaken
for 5 hr. The mixture was filtered and the filtrate was
concentrated in vacuo. The residue was purified by Sep-Pak column.
After removal of the solvent, Et.sub.2O was added to the residue
and the solid was collected to afford 8 mg 9 as a crystalline
solid. MS (FAB) m/z 504 (H+1)
[0304] To the above resin (637 mg) was added a solution of
piperidine in DMF (50% v/v, 20 mL) and the mixture was shaken for 4
hr. The resin was washed with DMF (.times.3), MeOH (.times.3),
CH.sub.2Cl.sub.2 (.times.3). To the resin was added DMF (12 mL),
CH.sub.2Cl.sub.2 (8 mL), 4-(Fmoc-amino)phenylacetic acid (530 mg,
1.42 mmol), PyBrop (660 mg, 1.43 mmol) and DIEA (0.62 mL, 3.56
mmol). The mixture was shaken for 60 hr and drained. The resin was
washed with DMF (.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2
(.times.3) and dried under a reduced pressure to afford 617 mg of
the resin. 57 mg of this resin was added Piperidine in DMF (40%
v/v, 2 mL). The mixture was shaken for 1 hr. The resin was washed
with DMF (.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2 (.times.3).
2-chlorophenyl isocyanate (0.050 mL, 0.41 mmol) was added to a
suspension of resin in THF (1 mL) and CH.sub.2Cl.sub.2 (1 mL). The
mixture was shaken for 20 hr and drained. The resin was washed with
DMF (.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2 (.times.3). To
the resin was added a solution of TFA in CH.sub.2Cl.sub.2 (25% v/v,
2 mL) and the mixture was shaken for 1.5 hr. The mixture was
filtered and the filtrate was concentrated in vacuo. The residue
was purified by Sep-Pak column. After removal of the solvent,
Et.sub.2O was added to the residue and the solid was collected to
afford 2 mg 10 as a crystalline solid. MS (FAB) m/z 508
(M.sup.++1)
Example 8
(S)-3-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]pyrrolidinyl]methox-
y]phenylacetic acid
[0305] ##STR1632##
(S)-3-[2-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]pyrrolidinyl]methox-
y]phenylacetic acid
[0306] ##STR1633##
[0307] 11 and 12 To a stirred mixture of Boc-prolinol (3.51 g, 17.5
mmol), methyl m-hydroxyphenylacetate (2.90 g, 17.5 mmol),
triphenylphosphine (4.60 g, 17.6 mol) in THF (50 mL) was added
dropwise diethyl azodicarboxylate (3.05 g, 17.5 mmol) at room temp.
After the addition was completed, the mixture was heated under
reflux for 3 hr. After cooling, the mixture was concentrated in
vacuo. The residue was dissolved in EtOAc, washed successively with
1 N NaOH, water, brine and dried over MgSO.sub.4. After removal of
the solvent, the residue was purified by column chromatography on
silica-gel with EtOAc-hexane (1:4, v/v) as eluent to give 5.49 g
(90%) methyl
(S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxyphenylacetate as
an oil.
[0308] A mixture of the above methyl
(S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy phenylacetate
in MeOH (60 mL) and 1 N NaOH (20 mL) was stirred for 8 hr at room
temp. After removal of the solvent under a reduced pressure, water
(50 mL) was added to the residue. The mixture was extracted with
Et.sub.2O (.times.2), and the aqueous layer was acidified by the
addition of 1 N HCl. The mixture was extracted with EtOAc. The
extract was washed with water, brine, dried over MgSO.sub.4 and
then concentrated in vacuo to afford 4.43 g (88%)
(S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy phenylacetic
acid as a viscous oil.
[0309] A mixture of the above
(S)-3-(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxyphenyl acetic
acid in CH.sub.2Cl.sub.2 (10 mL) and TFA (10 mL) was stirred for 1
hr at room temp. Et.sub.2O was added to the mixture and allowed to
stand. Upper layer was removed by decantation to give an oil. A
mixture of this oil in water (100 mL), dioxane (30 mL) and
NaHCO.sub.3 (6.0 g) was added Fmoc-Cl (2.86 g, 11.1 mmol) and the
mixture was stirred for 20 hr at room temp. The mixture was
extracted with Et.sub.2O (.times.2), and the aqueous layer was
acidified by the addition of 1 N HCl. The mixture was extracted
with EtOAc. The extract was washed with water, brine, dried over
MgSO.sub.4 and concentrated in vacuo to afford 5.08 g (81%)
(S)-3-(1-Fmoc-2-pyrrolidinyl)methoxyphenylacetic acid as a viscous
oil.
[0310] Wang resin (0.71 mmol/g, 400 mg) was suspended in a solution
of (S)-3-(1-Fmoc-2-pyrrolidinyl)methoxyphenylacetic acid (520 mg,
1.14 mol), DMAP (35 mg, 0.29 mmol), HOBt (40 mg, 0.30 mmol) and DIC
(0.45 mL, 2.9 mmol) in a mixture of DMF (3 mL) and CH.sub.2Cl.sub.2
(7 mL). The mixture was shaken for 20 hr and drained. The resin was
washed with DMF (.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2
(.times.3) and dried under a reduced pressure to give 593 mg of
resin, which was used for the preparation of 11 and 12.
[0311] 11 A mixture of the above resin (70 mg) in piperidine-DMF
(40% v/v, 3 mL) was shaken for 1 hr. The resin was washed with DMF
(.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2 (.times.3). To the
resin was added DMF (1.5 mL), CH.sub.2Cl.sub.2 (1.5 mL) 3-methoxy-4
(N'-phenylureido)phenylacetic acid (42 mg, 0.14 mmol), PyBrop (70
mg, 0.15 mmol) and DIEA (0.065 mL, 0.37 mmol). The mixture was
shaken for 15 hr and drained. The resin was washed with DMF
(.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2 (.times.3). To the
resin was added a solution of TFA in CH.sub.2Cl.sub.2 (25% v/v, 2
mL) and the mixture was shaken for 3 hr. The mixture was filtered
and the filtrate was concentrated in vacuo. The residue was
purified by Sep-Pak column. After removal of the solvent, Et.sub.2O
was added to the residue and the solid was collected to afford 8 mg
11 as a crystalline solid. MS (FAB) m/z 518 (M.sup.++1)
[0312] 12 A mixture of the above resin (70 mg) in piperidine-DMF
(40% v/v, 3 mL) was shaken for 1 hr. The resin was washed with DMF
(.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2 (.times.3). To the
resin was added DMF (1.5 mL), CH.sub.2Cl.sub.2 (1.5 mL),
4-[N'-(2-methylphenyl)ureido]phenylacetic acid (40 mg, 0.14 mmol),
PyBrop (70 mg, 0.15 mmol) and DIEA (0.065 mL, 0.37 mmol). The
mixture was shaken for 15 hr and drained. The resin was washed with
DMF (.times.3), MeOH (.times.3), CH.sub.2Cl.sub.2 (.times.3). A
mixture of the resin in TFA-CH.sub.2Cl.sub.2 (25% v/v, 2 mL) was
shaken for 3 hr. The mixture was filtered and the filtrate was
concentrated in vacuo. The residue was purified by Sep-Pak column.
After removal of the solvent, Et.sub.2O was added to the residue
and the solid was collected to afford 11 mg 12 as a crystalline
solid. MS (FAB) m/z 502 (M.sup.++1)
Example 9
4-[2-[1-[3-methoxy-4N'-phenylureido)phenylacetyl]-2-pyrrolidinyl]ethynyl]b-
enzoic acid
[0313] ##STR1634##
[0314] To a stirred cold (minus 50.degree. C.) solution of
N-boc-prolinal (5.98 g, 30 mmol) and PPh.sub.3 (62.95 g, 240 mmol)
in CH.sub.2Cl.sub.2 (200 mL) was slowly added a solution of
CBr.sub.4 (39.80 g, 120 mmol) in CH.sub.2Cl.sub.2 (50 mL), and the
stirring was continued for 1 hr at 0.degree. C. To this mixture was
added sat. NaHCO.sub.3 and the mixture was extracted with
CHCl.sub.3. The extract was washed with H.sub.2O, dried over
MgSO.sub.4, and evaporated. The residue was chromatographed on
silica-gel with CHCl.sub.3 and n-hexane-AcOEt (4:1, v/v) as eluent
to give 7.84 g (74%)
1-(tert-butoxycarbonyl)-2-(2,2-dibromoethenyl)pyrrolidine as
colorless plates. mp 61-63; IR (KBr) 1693 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) 37.46 (9H, s), 1.72-2.19 (4H, m), 3.35-3.45 (2H, m),
4.35 (1H, br s), 6.36 (1H, br s); MS (FAB) m/z 352, 354, 356, 358;
Anal. Calcd for C.sub.11H.sub.17NO.sub.2Br.sub.2: C, 37.21; H,
4.83; N, 3.94. Found: C, 37.14; H, 4.83; N, 4.00.
[0315] To a stirred cold (minus 78.degree. C.) solution of
1-(tert-butoxycarbonyl)-2-(2,2-dibromoethenyl)pyrrolidine (7.81 g,
22 mmol) in THF (200 mL) was added n-BuLi (1.59 M in hexane, 28 mL,
44 mmol) over 10 min, and the stirring was continued for 2 hr at
the same temp. The reaction was quenched by the addition of sat.
NH.sub.4Cl and extracted with EtOAc. The extract was washed with
brine, dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with n-hexane-AcOEt (10:1, v/v) as
eluent to give 4.15 g (97%) 1-(tert-butoxycarbonyl)-2-ethynyl
pyrrolidine as a light yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.48 (9H, s), 1.82-2.21 (4H, m), 3.30-3.45 (2H, m), 4.41-4.52 (1H,
m).
[0316] A suspension of ethyl 4-iodobenzoate (1.7 mL, 10 mmol),
Pd(PPh.sub.3).sub.4 (578 mg, 0.5 mmol), and CuI (190 mmol, 1 mmol)
in i-Pr.sub.2NH (20 mL) was stirred for 0.5 hr under N.sub.2. To
this mixture was added a solution of
1-(tert-butoxycarbonyl)-2-ethynylpyrrolidine (1.95 g, 10 mmol) in
i-Pr.sub.2NH (20 mL) for over 10 min. After stirring for 3 hr at
room temp, the mixture was poured into H.sub.2O and extracted with
EtOAc. The extract was washed with brine, dried over MgSO.sub.4,
and evaporated. The residue was chromatographed on silica-gel with
n-hexane-AcOEt (10:1, v/v) as eluent to give 2.77 g (81%)
1-(tert-butyloxycarbonyl)-2-(2-(4-ethoxycarbonylphenyl)ethynyl)pyrrolidin-
e as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.37 (3H, t,
J=6.8 Hz), 1.49 (9H, s), 1.85-2.12 (4H, m), 3.37-3.51 (2H, m), 4.37
(2H, q, J=6.8 Hz), 4.54-4.77 (1H, m), 7.44 (2H, d, J=7.8 Hz), 7.96
(2H, d, J=7.8 Hz)
[0317] To a stirred solution of
1-(tert-butoxycarbonyl)-2-[2-(4-ethoxycarbonylphenyl)ethynyl]pyrrolidine
(2.75 g, 8 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added TFA (5 mL),
and the resulting mixture was stirred overnight. The mixture was
concentrated in vacuo and made basic with sat. NaHCO.sub.3 and
extracted with CHCl.sub.3. The extract was washed with brine, dried
over MgSO.sub.4, evaporated to give 1.95 g (q.y.)
2-[2-(4-ethoxycarbonylphenyl)ethynyl]pyrrolidine as a light yellow
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.38 (3H, t, J=6.8 Hz),
1.82-2.16 (4H, m), 3.01-3.48 (2H, m), 4.00-4.11 (1H, m), 4.37 (2H,
q, J=6.8 Hz), 4.54-4.77 (1H, m), 7.44-7.46 (2H, m), 7.95-7.97 (2H,
m).
[0318] A mixture of 3-methoxy-4-(N'-phenylureido)phenylacetic acid
(180 mg, 0.6 mmol),
2-(2-(4-ethoxycarbonylphenyl)ethynyl)pyrrolidine (146 mg, 0.6
mmol), EDC (173 mg, 0.9 mmol), DMAP (73 mg, 0.6 mmol), and cat.
HOBt in DMF (10 mL) was stirred overnight. The mixture was poured
into 1 N HCl and the solid was collected with suction. The residue
was dissolved in CHCl.sub.3 and dried over MgSO.sub.4. After
removal of the solvent, the residue was chromatographed on
silica-gel with CHCl.sub.3-MeOH (10:1, v/v) as eluent to give 192
mg (61%) ethyl
4-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]-2-pyrrolidinyl]ethyny-
l]benzoate as a light yellow amorphous solid. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.38 (3H, t, J=6.8 Hz), 1.98-2.24 (4H, m),
3.48-3.89 (2H, m), 3.53 (2H, s), 3.62 (3H, s), 4.33-4.40 (2H, m),
4.78-5.04 (1H, m), 6.77-8.00 (14H, m).
[0319] To a stirred solution of ethyl
4-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]-2-pyrrolidinyl]ethyny-
l]benzoate (184 mg, 0.35 mmol) in THF (5 mL) was added 0.25 N NaOH
(4 mL). The resulting mixture was stirred overnight. The mixture
was poured into H.sub.2O and made acidic by the addition of 1 N HCl
(1 mL). The solid was collected with suction and dissolved in
CHCl.sub.3. The solution was dried over MgSO.sub.4 and evaporated.
The residue was recrystallized from CHCl.sub.3-n-hexane to give 65
mg (37%) 13 as a white crystalline powder. mp 154-157; IR (KBr)
3346, 2952, 2615, 1712, 1693 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.92-2.29 (4H, m), 3.32-3.82 (2H, m), 3.78 (2H, s), 3.80
(3H, s), 4.87-5.11 (1H m), 6.77-9.26 (14H, m), 13.10 (1H, br s); MS
(FAB) m/z 498 (M.sup.++1).
Example 10
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
yl]ethynyl]benzoic acid
[0320] ##STR1635##
[0321] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.45 g,
4.6 mmol), 2-(2-(4-ethoxycarbonylphenyl)ethynyl)pyrrolidine (1.12
g, 4.6 mmol), EDC 1.32 g (6.9 mmol), DMAP (562 mg, 4.6 mmol) in DMF
(20 mL) was stirred overnight The mixture was poured into 1 N HCl
and the solid was collected with suction. The solid was dissolved
in CHCl.sub.3 and dried over MgSO.sub.4. After removal of the
solvent, the residue was chromatographed on silica-gel with
CHCl.sub.3-MeOH (100:1, v/v) as eluent to give 2.20 g (89%) ethyl
4-[2-[1-[3-methoxy-4-(N'-2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
yl]ethynyl]benzoate as a white amorphous solid.
[0322] .sup.1H-NMR (CDCl.sub.3) .delta. 1.37-1.41 (3H, m),
1.94-2.22 (4H, m), 2.29 (3H, s), 3.41-3.89 (2H, m), 3.62 (3H, s),
3.69 (2H, s), 4.34-4.40 (2H, m), 4.72-5.01 (1H, m), 6.33 (1H, br
s), 6.80-8.06 (12H, m).
[0323] To a stirred solution of ethyl
4-[2-[1-[3-methoxy-4-(N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl]ethynyl]benzoate (2.16 g, 4 mmol) in THF (30
mL) was added 0.25 N NaOH (32 mL) and the stirring was continued
overnight. The mixture was poured into H.sub.2O and acidified by
the addition of 1 N HCl (8 mL). The resulting precipitate was
collected with suction and dissolved in CHCl.sub.3. The solution
was dried over MgSO.sub.4 and evaporated. The residue was
recrystallized from CHCl.sub.3-n-hexane to give 555 mg (27%) 14 as
a white crystalline powder. mp 161-164; IR (KBr) 3338, 2954, 2875,
1707, 1691 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.96-2.10
(4H, m), 2.24 (3H, s), 3.32-3.81 (2H, m), 3.62 (2H, s), 3.81 (3H,
s), 4.87-5.10 (1H, m), 6.76-8.58 (13H, m); MS (FAB) m/z 512
(M.sup.++1)
Example 11
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
yl]ethynyl]phenylacetic acid
[0324] ##STR1636##
[0325] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (141 mg,
0.45 mmol), 2-[2-(3-ethoxycarbonylmethylphenyl)ethynyl]pyrrolidine
(116 mg, 0.45 mmol), EDC (130 mg, 0.68 mmol), DMAP (55 mg, 0.45
mmol), cat. HOBt in DMF (10 mL) was stirred overnight. The mixture
was poured into 1 N HCl and the solid was collected by filtration.
The solid was dissolved in CHCl.sub.3, dried over MgSO.sub.4, and
evaporated. The residue was chromatographed on silica-gel with
CHCl.sub.3-MeOH (100:1 v/v) as eluent to give an oil, which was
dissolved in THF (5 mL). 0.25 N NaOH (4 mL) was added to this
solution with stirring. After stirring overnight, the mixture was
poured into 1N HCl (20 mL). The resulting precipitate was collected
with suction and dissolved in CHCl.sub.3. The solution was dried
over MgSO.sub.4 and evaporated. The residue was chromatographed on
silica-gel with CHCl.sub.3-MeOH (5:1, v/v) as eluent to give 92 mg
(39%) 15 as a white amorphous solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.96-2.18 (7H, m), 3.50-3.88 (9H, m), 4.78-4.98 (2H, m),
6.72-7.99 (14H, m); MS (FAB) m/z 526 (M.sup.++1).
Example 12
4-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinylmethoxy]iso-
phthalic acid
[0326] ##STR1637##
[0327] To a stirred solution of pentafluorophenyl ester of
4-[N'-(2-methylphenyl)ureido]phenylacetic acid (2.32 g, 5.15 mmol),
dimethyl (S)-4-(2-pyrrolidinylmethoxy)isophthalate (1.51 g, 5.15
mmol) in DMF (20 mL) was added Et.sub.3N (1.0 mL, 6.65 mmol), and
the mixture was stirred overnight. The resulting mixture was
diluted with EtOAc. The solution was washed with brine, dried over
MgSO.sub.4, and evaporated off in vacuo. The residue was purified
by column chromatography on silica-gel with CHCl.sub.3-MeOH (10:1,
v/v) as eluent to give 1.58 g (55%) dimethyl
4-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinylmethoxy]is-
ophthalate as a yellow crystalline solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.87-2.25 (m, total 7H), 3.50-3.65 (m, 4H), 3.85 (s, 3H),
3.89 (s, 3H), 4.18-4.31 (m, 2H), 4.44 (m, 1H), 6.95-8.45 (m, total
13H).
[0328] To a stirred solution of dimethyl
4-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinylmethoxy]is-
ophthalate (1.56 g, 2.79 mmol) in THF (30 mL) was added 0.25 N NaOH
(20 mL), and the reaction mixture was heated under reflux
overnight. The resulting mixture was poured into 1 N HCl, and solid
was collected. The crude solid was washed with Et.sub.2O to give
574 mg (39%) 16 as a yellow amorphous solid. IR (KBr) 1710
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) 1.83-2.18 (m, 4H), 2.24 (s,
3H), 3.36-4.28 (m, 8H) 6.91-9.02 (series of m, total 13H), 12.89
(br s, 1H); MS (FAB) m/z 532 (M.sup.++1).
Example 13
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
yl]ethenyl]benzoic acid
[0329] ##STR1638##
[0330] To a stirred solution of the 4-[2-[2
(N-tert-butoxycarbonyl)pyrrolydinyl]ethenyl]benzonitrile (2.26 g,
7.57 mmol) in CH.sub.2Cl.sub.2 (23 mL) was added dropwise a 1.5M
solution of diisopropylaluminum hydride (toluene solution) (6.06
mL, 9.09 mmol) at 0.degree. C. for over 15 min. The resulting
solution was stirred for 3 hr at 0.degree. C. The solution was
quenched by the addition of sat.NH.sub.4Cl. The resulting mixture
was filtered through Celite, and the filtrate was extracted with
EtOAc. The filtrate was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated in vacuo to afford 1.89 g (83%)
4-[2-[2-(N-tert-butoxy carbonyl)pyrrolidinyl]ethenyl]benzaldehyde
as a yellow syrup.
[0331] To a stirred solution of NaOH (1.00 g, 25.1 mmol) in water
(10 mL) was added a solution of AgNO.sub.3 (2.13 g, 12.5 mmol) in
CH.sub.3CN (10 mL) at 0.degree. C. for over 0.5 hr. To the stirred
above mixture was added dropwise a solution of
4-[2-[2-(N-tert-butoxycarbonyl)pyrrolydinyl]ethenyl]benzaldehyde
(1.89 g, 6.27 mmol) in CH.sub.3CN (10 mL) at 0.degree. C. for over
20 min. After the resulting mixture was stirred for a further 3 hr
at room temp. The mixture was filtered with suction, and then
washed with hot water. After the filtrate was washed with EtOAc,
the aqueous layer was acidified by carefully adding 1 N HCl, and
then extracted with CHCl.sub.3. The extract was dried over
Na.sub.2SO.sub.4 and evaporated in vacuo to afford 0.700 g (35% for
2 steps)
4-[2-[2-(N-tert-butoxycarbonyl)pyrrolidinyl]ethenyl]benzoic acid as
a pale yellow crystalline material.
[0332] To a stirred solution of
4-[2-(2-(N-tert-butoxycarbonyl)pyrrolidinyl)ethenyl]benzoic acid
(0.700 g, 2.21 mmol) in MeOH-benzene (1:4, v/v, 30 mL) was added
dropwise a 2 M-n-hexane solution of TMSCHN.sub.2 (1.32 mL, 2.65
mmol) at room temp. After the solution was stirred for 0.5 hr at
room temp, the solution was evaporated in vacuo. The resulting oily
residue was chromatographed on silica-gel with EtOAc-n-hexane (1:6,
v/v) as eluent to afford 0.64 g (88%) methyl
4-[2-[2-(N-tert-butoxycarbonyl)pyrrolidinyl]ethenyl]benzoate as a
pale yellow crystalline material.
[0333] To a stirred solution of methyl
4-[2-[2-(N-tert-butoxycarbonyl)pyrrolidinyl]ethenyl]benzoate (0.64
g, 1.93 mmol) in CH.sub.2Cl.sub.2 (5 mL) was added TFA (5 mL) at
room temp. After the mixture was stirred for 1 hr at room temp, the
mixture was evaporated in vacuo, The residue was treated with sat.
NaHCO.sub.3 and extracted with CHCl.sub.3. The extract was dried
over Na.sub.2SO.sub.4 and evaporated in vacuo to afford 0.45 g
(100%) methyl 4-[2-(2-pyrrolidinyl)ethenyl]benzoate as a yellow
crystalline material.
[0334] To a stirred mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (285 mg,
0.906 mmol), methyl 4-[2-(2-pyrrolydinyl)ethenyl]benzoate (210 mg,
0.906 mmol) in DMF (4 mL) was added
1-ethyl-3-(3-methylaminopropyl)carbodiimide (EDC) (209 mg, 1.09
mmol), 1-hydroxybenzotriazole (HOBt) (147 mg, 1.09 mmol) and
4-dimethylaminopyridine (DMAP) (11 mg, 0.0906 mmol) at room temp.
After the resulting mixture was stirred for 48 hr at room temp, the
mixture was poured into ice-1 N HCl and extracted with EtOAc. The
extract was dried over Na.sub.2SO.sub.4 and evaporated in vacuo.
The residue was chromatographed on silica-gel with acetone-toluene
(1:4 to 1:1, v/v) as eluent to afford 0.47 g (98%) methyl
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nyl]ethenyl]benzoate as a white crystalline material.
[0335] To a stirred solution of methyl
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl]ethenyl]benzoate (0.47 g, 0.891 mmol) in THF
(5 mL) was added 0.25 N NaOH (5.36 mL). The resulting mixture was
stirred at room temp for 20 hr. The mixture was acidified with 1N
HCl. The mixture was extracted with EtOAc. The extract was washed
with brine, dried over Na.sub.2SO.sub.4 and evaporated in vacuo to
afford 430 mg (94%) 17 as a white crystalline material. .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 1.78-2.13 (4H, m), 2.50 (3H, s),
3.44-3.68 (4H, m), 3.75 and 3.82 (3H, s), 4.71 (1H, m), 6.26-8.59
(15H, m).
Example 14
4-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]-2-pyrrolidinyl]ethenyl-
]benzoic acid
[0336] ##STR1639##
[0337] To a stirred solution of
3-methoxy-4-(N'-phenylureido)phenylacetic acid (305 mg, 1.01 mmol),
methyl 4-[2-(2-pyrolidinyl)ethenyl]benzoate (235 mg, 1.01 mmol) in
DMF (4 mL) was added 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
(232 mg, 1.21 mmol), 1-hydroxybenzotriazole (HOBt) (164 mg, 1.21
mmol), and 4-dimethylaminopyridine (DMAP) (12 mg, 0.101 mmol) at
room temp. After the mixture was stirred for 48 hr, the mixture was
acidified by the addition of 1 N HCl. The mixture was extracted
with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated in vacuo to give an oily residue.
The residue was chromatographed on silica-gel with acetone:toluene
(1:4 to 1:1, v/v) as eluent to afford 0.43 g (83%) methyl
4-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]-2-pyrrolidinyl]etheny-
l]benzoate acid as a white crystalline material.
[0338] To a stirred solution of methyl
4-[2-[1-[3-methoxy-4-(N'-phenylureido)phenylacetyl]-2-pyrrolidinyl]etheny-
l]benzoate (0.43 g, 0.837 mmol) in THF (5 mL) was added a solution
of 0.25 N NaOH (5.04 mL) at room temp. After the resulting mixture
was stirred for 20 hr, the mixture was acidified by the carefully
addition of 1 N HCl. The mixture was extracted with EtOAc. The
extract was washed with brine, dried over Na.sub.2SO.sub.4 and
evaporated in vacuo to afford 397 mg (95%) 18 as a white
crystalline material. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
1.78-2.13 (4H, m), 3.17-3.68 (4H, m), 3.74, 3.82 (3H, s), 4.71 (1H,
m), 6.27-9.28 (16H, m).
Example 15
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
yl]ethyl]benzoic acid
[0339] ##STR1640##
[0340] A mixture of
4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nyl]ethenyl]benzoic acid (184 mg, 0.358 mmol) and 5% Pd--C(368 mg)
in MeOH was hydrogenated in an atmospheric pressure at room temp.
After the mixture was stirred for 21 hr at room temp, insoluble
catalyst was filtered off and the filtrate was evaporated in vacuo.
The residue was chromatographed on silica-gel with MeOH--CHCl.sub.3
(1:4 to 1:3, v/v) as eluent to afford 123 mg (66%) 19 as a white
crystalline material. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.55-2.03
(m, 6H), 2.24 (s, 3H), 2.60 (m, 2H), 3.17-3.59 (m, 4H), 3.83 (s,
3H), 3.97 (m, 1H), 6.61-8.57 (m, 13H); MS (FAB) m/z 516
(M.sup.++1).
Example 16
3-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]-
methylthiobenzoic acid
[0341] ##STR1641##
[0342] To a solution of m-iodophenol (20.0 g, 90.9 mmol) in DMF
(200 mL) was added 1,4-diazabicyclo[2,2,2]octane (20.4 g, 181.8
mmol) and dimethylthiocarbamoyl chloride (16.9 g, 136.4 mmol). The
resulting cloudy solution was stirred for 0.5 hr at 35.degree. C.
and then heated at 75.degree. C. for 0.5 hr. After cooling, 300 mL
of water was added to the mixture. The solid was collected, washed
with water and dried under a reduced pressure to give 27.63 g (99%)
O-m-iodophenyl dimethylthio carbamate as a pale yellow crystalline
powder. IR (KBr) 1540, 1463, 1278, 1193, 1166, 1124 cm.sup.-1;
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 3.33 (s, 3H), 3.45 (s,
3H), 7.05-7.14 (m, 2H), 7.43 (d, J=1.9 Hz, 1H), 7.58 (dd, J=1.0,
7.8 Hz, 1H); MS (FAB) m/z 307 (M.sup.++1); Anal. Calcd for
C.sub.9H.sub.10INOS: C, 35.19; H, 3.28; N, 4.56. Found: C, 35.23;
H, 3.40; N, 4.41.
[0343] A solution of O-m-iodophenyl dimethylthiocarbamate (10.0 g,
32.6 mmol) in Ph.sub.2O (25 mL) was heated at 230.degree. C. for 10
hr. After cooling, the reaction mixture was chromatographed on
silica-gel with n-hexane-EtOAc (5:1, v/v) as eluent to give 9.31 g
(93%) S-m-iodophenyl dimethylthio carbamate as a pale yellow oil.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 3.08 (br s, 6H), 7.11 (t,
J=7.8 Hz, 1H), 7.46 (d, J=7.3 Hz, 1H), 7.71 (d, J=7.3 Hz, 1H), 7.85
(s, 1H); MS (FAB) m/z 307 (M.sup.++1).
[0344] To a solution of S-m-iodophenyl dimethylthiocarbamate (5.01
g, 16.31 mmol) in MeOH (20 mL) was added 28%-MeONa in MeOH (3.46
mL, 17.94 mmol). The resulting mixture was stirred at room temp for
3.5 hr and then heated at 70.degree. C. overnight. After cooling,
1N HCl was added. The solvent was removed under a reduced pressure
and the residue was diluted with EtOAc. The solution was washed
with H.sub.2O, brine, and dried over Na.sub.2SO.sub.4. The organic
layer was concentrated under a reduced pressure. The residue was
chromatographed on silica-gel with n-hexane-AcOEt (10:1, v/v) as
eluent to afford 3.42 g (89%) m-iodothiophenol as an oil.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 3.45 (s, 1H), 6.95 (t,
J=7.8 Hz, 1H), 7.23 (d, J=7.8 Hz 1H), 7.48 (d, J=7.3 Hz, 1H), 7.64
(t, J=1.5 Hz, 1H); MS (EI) m/z 236 (M.sup.+).
[0345] To a stirred solution of
N-(tert-butoxycarbonyl)-2-pyrrolidinylmethanol (4.30 g, 20.0 mmol)
in pyridine (40 mL) was added p-TsCl (5.72 g, 30.0 mmol). The
resulting mixture was stirred at room temp for 3 hr. The reaction
mixture was quenched with H.sub.2O, and evaporated off. The residue
was diluted with EtOAc and washed with 1N HCl, brine, and dried
over Na.sub.2SO.sub.4. The solvent was removed under a reduced
pressure and the residue was chromatographed on silica-gel with
n-hexane-EtOAc (2:1, v/v) as eluent to afford 5.76 g (81%)
N-(tert-butoxycarbonyl)-2-pyrrolidinylmethyl p-toluenesulfonate as
a colorless oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.36 and
1.41 (s each, total 9H), 1.82 (br m, 2H), 1.96 (br m, 2H), 2.44 (s,
3H), 3.30 (br m, 2H), 3.89 (br s, 1H), 3.96 (br s, 1H), 4.09 (br m,
1H), 7.34 (br s, 2H), 7.77 (d, J=8.3 Hz, 2H); MS (FAB) m/z 356
(M.sup.++1).
[0346] To a stirred mixture of m-iodothiophenol (2.67 g, 11.31
mmol) and N-(tert-butoxy carbonyl)-2-pyrrolidinylmethyl
p-toluenesulfonate (3.34 g, 9.43 mmol) in pyridine (9.4 mL) was
added 8N KOH (1.77 mL). The resulting mixture was stirred at room
temp overnight. The reaction mixture was diluted with EtOAc. The
solution was washed with H.sub.2O, sat. NH.sub.4Cl solution, brine,
and dried over Na.sub.2SO.sub.1. The organic layer was concentrated
under a reduced pressure and the residue was chromatographed on
silica-gel with n-hexane-EtOAc (5:1, v/v) as eluent to afford 1.79
g (45%) [N-(tert-butoxycarbonyl)-2-pyrrolidinyl]methyl 3-iodophenyl
sulfide as an oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.45
(s, 9H), 1.78-2.01 (br m, 4H), 2.71 (dt, 1H), 3.32-3.49 (br m, 3H),
3.90-4.02 (br m, 1H), 7.12 (d, J=7.8 Hz, 1H), 7.18 (d, J=7.8 Hz,
1H), 7.57 (dd, J=2.0, 8.3 Hz, 2H); MS (FAB) m/z 420
(M.sup.++1).
[0347] To a stirred solution of
[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methyl 3-iodophenyl sulfide
(1.76 g, 4.20 mmol) in DMSO (20 mL) and MeOH (16 mL) was added
Et.sub.3N (1.28 mL, 9.24 mmol), Pd(OAc).sub.2 (47.1 mg, 0.21 mmol),
and 1,3-bis(diphenylphosphino)propane (86.6 mg, 0.2 mmol), then CO
gas was bubbled for 5 min. The resulting mixture was stirred at
70.degree. C. overnight. After cooling, the mixture was
concentrated. The residue was diluted with EtOAc and washed with
brine, and dried over Na.sub.2SO.sub.4. The solvent was removed
under a reduced pressure and the residue was chromatographed on
silica-gel with n-hexane-EtOAc (5:1, v/v) as eluent to afford 1.28
g (87%) methyl
3-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methylthiobenzoate as an
oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.42 and 1.45 (each
s, 9H), 1.79-2.05 (br m, 4H) 2.83 (dt, J=10.8, 30.3 Hz, 1H),
3.34-3.54 (br m, 3H), 3.92 (s, 3H), 3.92 and 4.05 (d, J=7.8 Hz,
1H), 7.36 (t, J=7.8 Hz, 1H), 7.63 (br d, J=14.7 Hz, 1H), 7.83 (br
d, J=12.7 Hz, 1H), 8.04 (s, 1H); MS (FAB) m/z 352 (M.sup.++1).
[0348] To a stirred solution of methyl
3-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methylthio benzoate (1.46
g, 4.16 mmol) in CH.sub.2Cl.sub.2 (30 mL) was added TFA (15 mL) at
0.degree. C. The resulting mixture was stirred at room temp for 1
hr. The solvent was removed under a reduced pressure and the
residue was treated with 1N NaOH and extracted with CHCl.sub.3. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated under a reduced pressure to afford 947 mg (91%) methyl
3-(2-pyrrolidinyl)methylthio benzoate as a brown oil. .sup.1H-NMR
(400 MHz, CDCl.sub.3) .delta. 1.45-1.54 (m, 1H), 1.72-2.00 (m, 4H)
2.88-3.10 (m, 4H), 3.30 (m, 1H), 3.92 (s, 3H), 7.34 (t, J=7.8 Hz,
1H), 7.52 (m, 1H), 7.84 (m, 1H), 8.01 (t, J=2.0 Hz, 1H); MS (FAB)
m/z 252 (M.sup.++1).
[0349] The mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.18 g,
3.77 mmol), EDC (1.08 g, 5.65 mmol), DMAP (23 mg, 0.19 mmol) and
HOBt (25 mg, 0.19 mmol) in DMF (5 mL) was stirred at room temp for
1 hr methyl 3-(2-pyrrolidinyl)methylthio benzoate (947 mg, 3.77
mmol) was added to the mixture and the resulting mixture was
stirred overnight. After DMAP (460 mg, 3.77 mmol) and HOBt (835 mg,
6.18 mmol) was added and stirred for a further 5 hr. The reaction
mixture was diluted with EtOAc. The solution was washed with brine
and dried over Na.sub.2SO.sub.4. The solvent was removed under a
reduced pressure. The residue was chromatographed on silica-gel
with n-hexane-EtOAc (2:3, v/v) a eluent to afford 294.3 mg (14%)
methyl
3-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylthio benzoate as a pale yellowish amorphous. IR (KBr) 2875,
1724, 1620, 1284, 1182 cm.sup.-1; .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. 1.86-2.05 (m, 4H), 2.31 (s, 3H), 2.84 (dd, J=9.3, 13.7 Hz,
1H), 3.43-3.59 (m, 5H), 3.73 (s, 3H), 3.92 (s, 3H), 4.33 (m, 1H),
6.16 (s, 1H), 6.77-6.80 (m, 2H), 7.04 (s, 1H), 7.16 (t, J=8.3 Hz,
1H), 7.38 (t, J=7.8 Hz, 1H), 7.49 (t, J=7.8 Hz, 1H), 7.73 (dt,
J=1.0, 7.8 Hz, 1H), 7.79 (dd, J=2.0, 6.8 Hz, 1H), 8.01 (d, J=2.0
Hz, 1H), 8.05 (dd, J=2.4, 7.8 Hz, 1H); MS (FAB) m/z 548
(M.sup.++1).
[0350] To a stirred solution of methyl
3-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylthiobenzoate (148.2 mg, 0.271 mmol) in THF (7.4 mL) and
H.sub.2O (1.8 mL) was added LiOH (19.4 mg, 0.812 mmol), and the
resulting mixture was stirred for 9 hr at room temp. The mixture
was treated with 1N HCl and extracted with CHCl.sub.3. The extract
was washed with brine and dried over Na.sub.2SO.sub.4. The solvent
was removed under a reduced pressure and the residue was purified
by preparative TLC eluting with CHCl.sub.3-MeOH (10:1, v/v), and
crystallized from n-hexane-EtOAc to afford 89.7 mg (62%) 20 as a
white powder. IR (KBr) 2960, 1708 cm.sup.-1; .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.82-2.01 (m, 4H), 2.24 (s, 3H), 2.93 (dd,
J=9.3, 12.7 Hz, 1H), 3.40-3.54 (m, 5H), 3.86 (s, 3H), 4.13 (br m,
1H), 6.74 (d, J=8.3 Hz, 1H), 6.87 (d, J=1.5 Hz, 1H), 6.94 (t, J=7.8
Hz, 1H), 7.10-7.17 (m, 2H), 7.42 (t, J=7.8 Hz, 1H), 7.70 (d, J=7.8
Hz, 1H), 7.74 (d, J=8.3 Hz, 1H), 7.79 (d, J=7.8 Hz, 1H), 7.84 (s,
1H), 8.00 (d, J=8.3 Hz, 1H), 8.48 (s, 1H), 8.57 (s, 1H); MS (FAB)
m/z 534 (M.sup.++1).
Example 17
3-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylsulfonyl]benzoic acid
[0351] ##STR1642##
[0352] To a stirred solution of methyl
3-[[1-[3-methoxy-4-[N'-(2-methyl phenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl]methylthio]benzoate (131.8 mg, 0.241 mmol)
in CH.sub.2Cl.sub.2 (3 mL) was added m-CPBA (130.5 mg, 0.529 mmol)
at 0.degree. C. The reaction mixture was stirred at room temp for
0.5 hr. The mixture was diluted with CHCl.sub.3, and quenched with
sat. Na.sub.2S.sub.2O.sub.3 solution The organic layer was
separated, washed with sat. NaHCO.sub.3 solution, brine, and dried
over Na.sub.2SO.sub.4. The solvent was removed under a reduced
pressure to afford methyl
3-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl]-methylsulfonyl]benzoate as an amorphous
solid. To a stirred solution of this crude compound in THF (7.4 mL)
and H.sub.2O (1.8 mL) was added LiOH (17.3 mg, 0.723 mmol), and the
stirring was continued overnight at room temp. The reaction mixture
was diluted with CHCl.sub.3 and washed with 1N HCl, then brine, and
dried over Na.sub.2SO.sub.4. The solvent was removed under a
reduced pressure and the residue was purified by preparative TLC
with CHCl.sub.3-MeOH (10:1, v/v) as eluent, and the crude solid was
recrystallized from n-hexane-EtOAc to afford 69.9 mg (51%) 21 as a
white crystalline powder. mp 243-245; IR (KBr) 3354, 2974, 1533
cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.80-2.00
(m, 4H), 2.24 (s, 3H), 3.19-3.62 (m, 6H), 3.82 (s, 3H), 4.18 (m,
1H), 6.67 (d, J=8.8 Hz, 1H), 6.80 (d, J=1.0 Hz, 1H), 6.93 (t, J=7.3
Hz, 1H), 7.10-7.17 (m, 2H), 7.66 (t, J=7.8 Hz, 1H), 7.78 (d, J=8.3
Hz, 1H), 7.91-7.99 (m, 2H), 8.20 (d, J=7.3 Hz, 1H), 8.34 (s, 1H),
8.48 (s, 1H), 8.56 (s, 1H); MS (FAB) m/z 566 (M.sup.++1); Anal.
Calcd for C.sub.29H.sub.31N.sub.3O.sub.7S.1HCl.1H.sub.2O: C, 56.17;
H, 5.53; N, 6.78. Found: C, 55.92; H, 5.58; N, 6.71.
Example 18
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylsulfonyl]benzoic acid
[0353] ##STR1643##
[0354] To a stirred solution of methyl
4-[[1-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]-2-pyrrolidinyl]methylthio]benzoate (300
mg, 0.548 mmol) in CH.sub.2Cl.sub.2 (6 mL) was added m-CPBA (297
mg, 1.206 mmol) at 0.degree. C., and the reaction mixture was
stirred at room temp for 1 hr. The mixture was diluted with
CHCl.sub.3, and quenched with sat. Na.sub.2S.sub.2O.sub.3 solution.
The separated organic layer was washed with sat. NaHCO.sub.3
solution, brine, and dried over Na.sub.2SO.sub.4. The solvent was
removed under a reduced pressure to afford methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]-methylsulfonyl]benzoate as a crude yellow oil. To a stirred
solution of this crude compound in THF (4.4 mL) and H.sub.2O (1.1
mL) was added LiOH (39.4 mg, 1.643 mmol), and the stirring was
continued overnight at room temp. The reaction mixture was diluted
with CHCl.sub.3 and washed with 1N HCl, brine, and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure
and the residue was purified by preparative TLC eluting with
CHCl.sub.3-MeOH (10:1, v/v), and crystallized from n-hexane-EtOAc
to afford 128.0 mg (41%) 22 as a white powder. IR (KBr) 3388, 2974,
1537, 1298, 1155 cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. 1.80-1.98 (m, 4H), 2.24 (s, 3H), 2.54 (s, 1H), 3.20-3.70
(m, 5H), 3.82 (s, 3H), 4.16 (br m, 1H), 6.67 (dd, J=1.5, 8.3 Hz,
1H), 6.80 (d, J=1.5 Hz, 1H), 6.93 (d, J=7.3 Hz, 1H), 7.10-7.16 (m,
2H), 7.78 (d, J=7.3 Hz, 1H), 7.95 (d, J=8.3 Hz, 2H), 7.98 (d, J=8.3
Hz, 1H), 8.14 (d, J=8.3 Hz, 2H), 8.49 (s, 1H), 8.57 (s, 1H); MS
(FAB) m/z 566 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.7S.3H.sub.2O: C, 56.21; H, 6.02; N,
6.78. Found: C, 56.76; H, 5.37; N, 6.70.
Example 19
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylthio]benzoic acid
[0355] ##STR1644##
[0356] To a stirred solution of p-iodophenol (20.0 g, 90.9 mmol) in
DMF (200 mL) was added 1,4-diazabicyclo[2,2,2]octane (20.4 g, 181.8
mmol) and dimethylthiocarbamoyl chloride (16.9 g, 136.4 mmol). The
resulting solution was stirred for 3.5 hr at 75.degree. C. After
cooling, 300 mL of water was added. The solid was collected with
suction and was dissolved in EtOAc. The EtOAc layer was washed with
water, dried over Na.sub.2SO.sub.4, and evaporated under a reduced
pressure. The crude solid was recrystallized from H.sub.2O to give
the 26.75 g (96%) O-p-iodophenyl dimethylthiocarbamate as a pale a
yellow crystalline powder. IR (KBr) 1479, 1207, 827 cm.sup.-1;
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 3.37 (s, 3H), 3.45 (s,
3H), 6.83 (d, J=8.8 Hz, 2H), 7.69 (d, J=8.3 Hz, 2H); MS (FAB) m/z
307 (M.sup.++1); Anal. Calcd for C.sub.9H.sub.10INOS: C, 35.19; H,
3.28; N, 4.56. Found: C, 35.17; H, 3.35; N, 4.44.
[0357] A stirred solution of O-p-iodophenyl dimethylthiocarbamate
(10.0 g, 32.6 mmol) in Ph.sub.2O (25 mL) was heated at 230.degree.
C. for 5.5 hr. After cooling, the reaction mixture was
chromatographed on silica-gel with n-hexane-EtOAc (3:1, v/v) as
eluent to give 2.55 g (26%) S-p-iodophenyl dimethylthio carbamate
as a white crystalline powder. IR (KBr) 3299, 1651, 1469, 1371
cm.sup.-1; .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 3.03 (br s,
3H), 3.08 (br s, 3H), 7.21 (d, J=8.3 Hz, 2H), 7.70 (d, J=8.3 Hz,
2H); MS (FAB) m/z 308 (M.sup.++1); Anal. Calcd for
C.sub.9H.sub.10INOS: C, 35.19; H, 3.28; N, 4.56. Found: C, 35.49;
H, 3.28; N, 4.43.
[0358] To a solution of S-p-iodophenyl dimethylthiocarbamate (2.55
g, 8.31 mmol) in MeOH (10 mL) was added MeONa (495 mg, 9.14 mmol),
and the resulting mixture was stirred at 70.degree. C. overnight.
After cooling, 1N HCl was added and the mixture was concentrated
under a reduced pressure. The residue was diluted with EtOAc and
washed with H.sub.2O, brine, and dried over Na.sub.2SO.sub.4. The
organic layer was concentrated under a reduced pressure and the
residue was chromatographed on silica-gel with n-hexane EtOAc (5:1,
v/v) as eluent to afford 1.75 g (89%) p-iodothiophenol as a pale
yellow crystalline solid. IR (KBr) 2559, 1097, 1002, 806 cm.sup.-1;
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 3.43 (s, 1H), 7.10 (d,
J=8.3 Hz, 2H), 7.53 (d, J=8.3 Hz, 2H); MS (FAB) m/z 236
(M.sup.++1); Anal. Calcd for C.sub.6H.sub.5IS: C, 30.53; H, 2.13.
Found: C, 30.57; H, 2.15.
[0359] To a stirred mixture of p-iodothiophenol (1.75 g, 7.43 mmol)
and N-(tert-butoxycarbonyl)-2-pyrrolidinylmethyl p-toluenesulfonate
(2.39 g, 6.75 mmol) in pyridine (12.7 mL) was added 8N KOH (1.27
mL) at room temp, and the resulting mixture was stirred for 4 hr at
the same temp. The reaction mixture was diluted with EtOAc. The
solution was washed with H.sub.2O, sat NH.sub.4Cl, brine, and dried
over Na.sub.2SO.sub.4. The organic layer was concentrated under a
reduced pressure. The residue was chromatographed on silica-gel
with n-hexane-EtOAc (5:1, v/v) as eluent to afford 1.49 g (53%)
[N-(tert-butoxycarbonyl)-2-pyrrolidinyl]methyl 4-iodophenyl sulfide
as a pale yellowish oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
(s, 9H), 1.78-2.01 (br m, 4H), 2.71 (dt, 1H), 3.32-3.49 (br m, 3H),
3.90-4.02 (br m, 1H), 7.12 (d, J=7.8 Hz, 1H), 7.18 (d, J=7.8 Hz,
1), 7.57 (dd, J=2.0, 8.3 Hz, 2H); MS (FAB) m/z 420 (M.sup.++1).
[0360] To a stirred solution of
[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methyl 4-iodophenyl sulfide
(1.49 g, 3.56 mmol) in DMSO (16 mL) and MeOH (13 mL) was added
Et.sub.3N (1.09 mL, 7.84 mmol), Pd(OAc).sub.2 (40 mg, 0.178 mmol),
and 1,3-bis(diphenylphosphino)propane (73.4 mg, 0.178 mmol). To the
stirred resulting mixture was induced CO gas for 5 min, and the
mixture was stirred at 70.degree. C. overnight. After cooling, the
mixture was concentrated to a small volume. The residue was diluted
with EtOAc, washed with brine, and dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure and the residue was
chromatographed on silica-gel with n-hexane-EtOAc (5:1, v/v) as
eluent to afford 1.16 g (93%) methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methylthiobenzoate as an
oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.51 and 1.47 (each
s, 9H), 1.78-2.05 (br m, 4H) 2.77 (dt, J=10.8, 37.1 Hz, 1H),
3.34-3.58 (m, 3H), 3.89 (s, 3H), 4.03 (br d, J=27.3 Hz, 1H), 7.38
(d, J=7.3 Hz, 1H), 7.47 (d, J=7.3 Hz, 1H), 7.92 (br s, 2H); MS
(FAB) m/z 352 (M.sup.++1).
[0361] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methylthio benzoate (1.16
g, 3.32 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added TFA (4 mL), and
the mixture was stirred at room temp for 1.5 hr. The solvent was
removed under a reduced pressure and the residue was treated with
1N NaOH, The mixture was extracted with CHCl.sub.3. The extract was
washed with brine, dried over KOH, and concentrated under a reduced
pressure to afford 767 mg (92%) methyl 4-(2-pyrrolidinyl)methylthio
benzoate as a yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
(dt, J=3.9, 12.7 Hz, 1H), 1.85-2.09 (m, 2H) 2.13 (m, 1H), 3.11-3.27
(m, 3H), 3.40 (dd, J=6.8, 13.2 Hz, 1H), 3.54 (dd, J=7.3, 15.1 Hz,
1H), 3.89 (s, 3H), 5.07 (br, 1H), 7.38 (d, J=8.3 Hz, 2H), 7.91 (d,
J=8.3 Hz, 2H); MS (FAB) m/z 252 (M.sup.+30 1).
[0362] To a stirred mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.30 g,
4.136 mmol), Et.sub.3N (0.63 mL, 4.549 mmol) in DMF (20 mL) was
added pentafluorophenyl trifluoroacetate at 0.degree. C. The
resulting mixture was stirred at room temp for 1 hr. The mixture
was poured into water (60 mL) and precipitate was collected with
suction. The crude solid was washed with 0.1N HCl, H.sub.2O,
n-hexane, and dried at 40.degree. C. to afford 1.91 g (96%)
pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate as a pale
brownish crystalline powder. IR (KBr) 1785, 1224, 1216 cm.sup.-1;
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 2.29 (s, 3H), 3.76 (s,
3H), 3.90 (s, 2H), 6.49 (s, 1H), 6.81 (d, J=1.5 Hz, 1H), 6.91 (dd,
J=1.5, 8.3 Hz, 1H), 7.15 (t, J=7.3 Hz, 3H), 7.24 (m, 1H), 7.50 (d,
J=7.8 Hz, 1H), 8.17 (d, J=7.8 Hz, 1H); MS (FAB) m/z 481
(M.sup.++1); Anal. Calcd for
C.sub.30H.sub.33N.sub.3O.sub.5S.1/4H.sub.2O: C, 57.51; H, 3.57; N,
5.83. Found: C, 57.40; H, 3.75; N, 5.68.
[0363] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (1.47 g, 3.05
mmol), methyl 4-(2-pyrrolidinyl)methylthiobenzoate (767 mg, 3.05
mmol), Et.sub.3N (0.51 mL, 3.66 mmol) in DMF (15 mL) was stirred
overnight at room temp. The reaction mixture was diluted with
EtOAc, washed with brine, and dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure and the residue was
chromatographed on silica-gel with n-hexane-EtOAc (1:2, v/v) as
eluent to afford 1.366 g (82%) methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methylthio]benzoate as a white crystalline powder. IR (KBr) 1785,
1224, 1216 cm.sup.-1; .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.88-1.99 (m, 4H), 2.30 (s, 3H), 2.75 (dd, J=9.8, 13.2 Hz, 1H),
3.43-3.55 (m, 3H), 3.56 (s, 2H), 3.64 (dd, J=1.1, 14.2 Hz, 1H),
3.73 (s, 3H), 3.88 (s, 3H), 4.33 (m, 1H), 6.29 (s, 1H), 6.78-6.81
(m, 2H), 7.11-7.26 (m, 5H), 7.50 (d, J=8.3 Hz, 3H), 7.93 (d, J=8.8
Hz, 2H), 8.07 (d, J=7.8 Hz, 1H); MS (FAB) m/z 548 (M.sup.++1);
Anal. Calcd for C.sub.30H.sub.33N.sub.3O.sub.5S.1/4H.sub.2O: C,
65.26; H, 6.12; N, 7.61. Found: C, 65.48; H, 6.20; N, 7.47.
[0364] To a stirred solution of methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methylthio]benzoate (300 mg, 0.548 mmol) in THF (5.5 mL) and
H.sub.2O (1.1 mL) was added LiOH (39.4 mg, 1.643 mmol), and the
reaction mixture was stirred at room temp overnight and at
50.degree. C. for 9 hr. The mixture was diluted with CHCl.sub.3.
The solution was washed with 1N HCl, brine, and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure,
and the obtained crude solid was recrystallized from
n-hexane-EtOAc-MeOH to afford 218.6 mg (75%) 23 as a white
crystalline powder. IR (KBr) 3318, 2952, 1596, 1536, 1299, 1155
cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.82-2.05
(m, 4H), 2.25 (s, 3H), 2.91 (dd, J=9.8, 13.2 Hz, 1H), 3.47-3.52 (m,
3H), 3.57 (s, 2H), 3.87 (s, 3H), 4.14 (br m, 1H), 6.76 (d, J=1.5,
8.3 Hzd, 1H), 6.89 (d, J=1.5 Hz, 1H), 6.94 (t, J=7.3 Hz, 1H),
7.11-7.19 (m, 2H), 7.57 (d, J=8.3 Hz, 2H), 7.80 (d, J=8.3 Hz, 1H),
7.83 (d, J=8.3 Hz, 2H), 8.02 (d, J=8.3 Hz, 1H), 8.49 (s, 1H), 8.58
(s, 1H); MS (FAB) m/z 534 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.5S. 5/4H.sub.2O: C, 62.63; H, 6.07; N,
7.36; S, 5.77. Found: C, 62.62; H, 5.74; N, 7.36; S, 5.67.
Example 20
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylsulfinyl]benzoic acid
[0365] ##STR1645##
[0366] To a stirred solution of methyl
4-[[1-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]-2-pyrrolidinyl]methylthio]benzoate (264
mg, 0.482 mmol) in CH.sub.2Cl.sub.2 (5.2 mL) was added m-CPBA
(118.8 mg, 0.482 mmol) at 0.degree. C., and the mixture was stirred
at room temp for 1 hr. The mixture was diluted with CHCl.sub.3, and
quenched with sat. Na.sub.2S.sub.2O.sub.3. The separated organic
layer was washed with sat. NaHCO.sub.3, brine, and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure
to afford methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methylsulfinyl]benzoate as a crude amorphous solid. To a stirred
solution of this crude compound in THF (4 mL) and H.sub.2O (1 mL)
was added LiOH (34.6 mg, 1.45 mmol), and the stirring was continued
overnight at room temp. The mixture was diluted with CHCl.sub.3,
washed with 1N HCl, brine, and dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure, and the obtained
crude solid was recrystallized from n-hexane-CHCl.sub.3-MeOH to
afford 193.2 mg (73%) 24 as a white crystalline powder. IR (KBr)
3338, 2956, 1708, 1529, 1299, 1207, 1155 cm.sup.-1; .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 1.70-2.06 (m, 4H), 2.24 (s, 3H),
2.90 (dd, J=8.3, 13.2 Hz, 1H), 3.02-3.08 (m, 1H), 3.16-3.25 (m,
1H), 3.41-3.60 (m, 3H), 3.84 (s, 3H), 4.40 (br s, 1H), 6.74 (d,
J=7.8 Hz, 1H), 6.87 (s, 1H), 6.94 (d, J=7.3 Hz, 1H), 7.11-7.17 (m,
2H), 7.75-7.81 (m, 3H), 7.98-8.05 (m, 1H), 8.10 (d, J=8.3 Hz, 2H),
8.46 (s, 1H), 8.56 (s, 1H); MS (FAB) m/z 550 (M.sup.++1), 572
(M.sup.++Na); Anal. Calcd for C.sub.29H.sub.31N.sub.3O.sub.6S.
3/2H.sub.2O: C, 60.40; H, 5.94; N, 7.29. Found: C, 60.15; H, 5.82;
N, 6.90.
Example 21
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacethyl]-2-pyrrolid-
inylmethoxy]benzoic acid
[0367] ##STR1646##
[0368] To a stirred solution of methyl 4-hydroxybenzoate (1.96 g,
12.88 mmol), N-Boc-prolinol (2.59 g, 12.87 mmol) and PPh.sub.3
(4.06 g, 15.48 mmol) in THF (40 mL) was added DIAD (3.10 mL, 15.74
mmol). The resulting mixture was heated under reflux for 14 hr. The
mixture was evaporated off in vacuo and the residue was purified by
column chromatography on silica-gel with n-hexane-EtOAc (6:1, v/v)
as eluent to give 3.34 g (77%) methyl
(S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethoxy]benzoate as an
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.48 (s, 9H), 1.67 (d, J=9.3
Hz, 1 H), 1.87-2.03 (m, 3H), 3.36-3.43 (m, 2H), 3.87-4.09 (m, 1H),
4.13-4.20 (m, 2H), 6.94 (d, J=8.3 Hz, 2H), 7.98 (d, J=8.3 Hz,
2H).
[0369] A mixture of methyl
(S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethoxy]benzoate (3.34
g, 9.96 mmol) in TFA (20 mL) and CH.sub.2Cl.sub.2 (35 mL) was
stirred at room temp for 15 hr. The mixture was concentrated in
vacuo and made basic with sat. NaHCO.sub.3. The mixture was
extracted with CHCl.sub.3, washed with brine, and dried over
Na.sub.2CO.sub.3. The organic layer was evaporated to give 1.70 g
(73%) methyl (S)-4-(2-pyrrolidinylmethoxy)benzoate as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.54-1.61 (m, 1H), 1.77-1.86 (m,
2H), 1.87-1.97 (m, 1H), 2.00 (bs, 1H), 2.93-3.06 (m, 2H), 3.52-3.57
(m, 1H), 3.88 (s, 3H), 3.90-3.99 (m, 2H), 6.92 (d, J=9.0 Hz, 2H),
7.98 (d, J=9.0 Hz, 2H)
[0370] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (428 mg,
1.36 mmol), methyl (S)-4-(2-pyrrolidinylmethoxy)benzoate (330 mg,
1.40 mmol), EDC (312 mg, 1.63 mg), HOBt (220 mg, 1.63 mmol), and a
catalytic amount of DMAP in DMF (15 mL) was stirred for 6 hr. The
resulting mixture was diluted with EtOAc, washed with 0.5 N HCl,
sat. NaHCO.sub.3, brine, and dried over MgSO.sub.4. The solvent was
evaporated off in vacuo to give an oily residue, which was purified
by column chromatography on silica-gel with CHCl.sub.3-MeOH (50:1,
v/v) as eluent to give 540 mg (75%) methyl
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolid-
inylmethoxy]benzoate as an oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.81-2.12 (m, 4H) 2.88 (bs, 3H), 3.48-3.61 (m, total 7H), 3.88 (s,
3H), 4.10-4.21 (m, 2H), 4.42-4.46 (m, 1H), 6.75-8.08 (series of m,
total 13H).
[0371] To a stirred solution of methyl
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolid-
inylmethoxy]benzoate (540 mg, 1.02 mmol) in THF (10 mL) was added
0.25 N NaOH (10 mL). The resulting mixture was heated under reflux
for 16 hr. The mixture was poured into 1 N HCl and the solid was
collected. The crude solid was washed with Et.sub.2O to give 278 mg
(53%) 25 as a white amorphous solid. IR (KBr) 1708 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.83-2.14 (m, 4H), 2.21 (s, 3H),
2.46 (s, 2H), 3.78 (s, 3H), 3.95-4.02 (m, H), 4.13-4.16 (m, 1H),
4.24 (bs, 1H), 6.51-7.98 (series of m, 12H), 8.43 (s, 1H), 8.53 (s,
1H), 12.57 (bs, 1H); MS (FAB) m/z 517 (M.sup.+).
Example 22
1-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]benzoyl]-L-prolyl]-4-piperidi-
nylacetic acid
[0372] ##STR1647##
[0373] A mixture of 3-methoxy-4-nitrobenzoic acid (229 mg, 1.16
mmol), tert-butyl 4-(1-prolylpiperidinyl)acetate (344 mg, 1.16
mmol), HOBt (188 mg, 1.39 mmol), DMAP (14.2 mg, 0.116 mmol), and
EDC (267 mg, 1.39 mmol) in DMF (7 mL) was stirred for 22 hr at room
temp. The mixture was diluted with EtOAc (50 mL) and washed
successively with 1 N HCl, sat. NaHCO.sub.3, and H.sub.2O. The
organic layer was dried over Na.sub.2SO.sub.4 and evaporated in
vacuo. The residue was chromatographed on silica-gel with
MeOH:CHCl.sub.3 (1:30, v/v) as eluent to afford 520 mg (94%)
tert-butyl
1-(3-methoxy-4-nitrobenzoyl)prolyl-4-(1-piperidinyl)acetate as a
white crystalline material. .sup.1H-NMR (CDCl.sub.3) .delta.
1.12-1.33 and 1.62-2.23 (each m, 9H), 1.44 (s, 9H), 2.65, 3.13,
3.47, 3.67, 4.44, and 4.61 (each m, 8H), 3.99 (s, 3H), 5.05 (m,
1H), 7.21 (d, J=8.3 Hz, 1H), 7.31 (s, 1H), 7.86 (d, J=8.3 Hz,
1H).
[0374] A stirred mixture of tert-butyl
1-(3-methoxy-4-nitrobenzoyl)prolyl-4-(1-piperidinyl)acetate (0.52
g, 1.09 mmol) and 5% Pd--C (2.08 g) in MeOH (10 mL) was
hydrogenated under an atmospheric pressure for 94 hr at room temp.
Insoluble catalyst was removed, and the filtrate was evaporated in
vacuo. The residue was chromatographed on silica-gel with
MeOH:CHCl.sub.3 (1:40 to 1:6, v/v) as eluent to afford 279 mg (57%)
tert-butyl
1-(4-amino-3-methoxybenzoyl)prolyl-4-(1-piperidinyl)acetate as a
white crystalline material. .sup.1H-NMR (CDCl.sub.3) .delta.
1.16-2.17, 2.69, 3.06, 3.67, 4.12, and 4.59 (each m, 17H), 3.86 (s,
3H), 5.10 (m, 1H), 6.64 (m, 1H), 7.12 (each m, 2H).
[0375] To a stirred solution of tert-butyl
1-(4-amino-3-methoxybenzoyl)-L-prolyl-4-(1-piperidinyl)acetate (279
mg, 0.627 mmol) and Et.sub.3N (0.0876 mL, 0.627 mmol) in THF (4 mL)
was added dropwise o-tolyl isocyanate (0.0777 mL, 0.627 mmol) at
room temp, and the resulting mixture was stirred for a further 21
hr at room temp. Ice water was added to the mixture and the
precipitate was collected with suction. The crude solid was
purified by silica-gel column chromatography with MeOH:CHCl.sub.3
(1:40, v/v) as eluent to afford 254 mg (70%) tert-butyl
1-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]benzoyl]-L-prolyl]-4-(1-pipe-
ridinyl)acetate as a crystalline solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.43 (s, 9H), 1.13-1.25 and 1.76-2.14 (each m, 9H), 2.60,
3.18, 3.71, 4.06, and 4.57 (each m, 8H), 3.67 (s, 3H), 5.06 (m,
1H), 6.63, and 6.90 (s, 2H), 6.98-7.23, and 7.64 (each m, 5H), 7.56
(d, J=7.8 Hz, 1H), 8.21 (d, J=8.8 Hz, 1H).
[0376] A solution of tert-butyl
1-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]benzoyl]prolyl]-4-(1-piperid-
inyl)acetate (254 mg, 0.440 mmol) in CH.sub.2Cl.sub.2 (6 mL) and
TFA (6 mL) was stirred for 5 hr at room temp. The mixture was
poured into ice water. The solid was collected with suction, washed
with water and air-dried to afford 179 mg (78%) 26 as a white
crystalline solid. .sup.1H-NMR (DMSO-d.sub.6) .delta. 0.47, 1.05,
1.44, and 1.62-1.99 (each m, 9H), 2.49 (s, 3H), 2.15-2.30, 2.35,
2.56, 2.78, 3.09, 3.04-3.80, 4.05, 4.15, and 4.32 (each m, 8H),
3.92 (s, 3H), 4.92 (m, 1H), 6.82, 6.95, 7.11, 7.77, 8.20, 8.57, and
8.75 (m, 9H); MS (FAB) m/z 523 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.34N.sub.4O.sub.6: C, 64.35; H, 6.56; N, 10.72. Found:
C, 55.58; H, 5.89; N, 8.75.
Example 23
(S)-3-methoxy-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-
-pyrrolidinyl methoxy]benzoic acid
[0377] ##STR1648##
[0378] To a stirred solution of ethyl 4-hydroxy-3-methoxybenzoate
(3.00 g, 15.29 mmol), (S)--N-Boc-prolinol (3.08 g, 15.30 mmol),
Ph.sub.3P (4.81 g, 18.34 mmol) in THF (50 mL) was added DIAD (3.61
mL, 18.33 mmol) at 0.degree. C. The resulting mixture was heated
under reflux 6.5 hr. After cooling to room temp, the mixture was
evaporated and purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (50:1, v/v) as eluent to give ethyl
(S)-3-methoxy-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl
methoxy]benzoate as a gum. The above ethyl
(S)-3-methoxy-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethoxy]benzoate
was dissolved in CH.sub.2Cl.sub.2 (50 mL) and TFA (45 mL). The
mixture was stirred for 2 days at room temp. The resulting mixture
was concentrated in vacuo and made basic with sat. NaHCO.sub.3. The
mixture was extracted with CH.sub.2Cl.sub.2, washed with brine, and
dried over MgSO.sub.4. The solvent was evaporated and the residue
was purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (20:1, v/v) as eluent to give 3.27 g (77% for 2
steps) ethyl (S)-3-methoxy-4-(2-pyrrolidinylmethoxy)benzoate as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.39 (t, 3H, J=7.1
Hz), 1.52-1.59 (m, 1H), 1.76-1.88 (m, 2H), 1.92-2.01 (m, 1H),
2.92-3.06 (m, 2H), 3.56-3.63 (m, 1H), 3.90 (s, 3H), 3.91-4.02 (m,
2H), 4.35 (q, 2H, J=7.1 Hz), 6.89 (d, 1H, J=8.3 Hz), 7.54 (d, 1H,
J=2.0 Hz), 7.65 (dd, 1H, J=2.0, 8.3 Hz).
[0379] To a stirred solution of ethyl
(S)-3-methoxy-4-(2-pyrrolidinylmethoxy)benzoate (424 mg, 1.52 mmol)
in DMF (8 mL) was added pentafluorophenyl ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (728 mg,
1.52 mmol) and Et.sub.3N (0.26 mL, 1.87 mmol). And the resulting
mixture was stirred at room temp overnight. The mixture was diluted
with EtOAc, washed with 1 N HCl, sat. NaHCO.sub.3, brine, and dried
over MgSO.sub.4. The solvent was evaporated off in vacuo and the
residue was purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (50:1, v/v) as eluent to give 830 mg (95%) ethyl
(S)-3-methoxy-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl methoxy]benzoate as an amorphous solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.38 (t, 3H, J=7.3 Hz), 1.88-2.20
(m, 4H, m), 2.24 (m, 3H), 3.44-3.50 (m, 1H), 3.53-3.58 (m, 7H),
3.82 (s, 3H); 4.09-4.17 (m, 1H), 4.22-4.25 (m, 1H), 4.35 (q, 2H,
J=7.3 Hz), 4.38-4.49 (m, 1H), 6.71-6.78 (m, 1H), 6.99 (d, 1H, J=8.3
Hz), 7.04-7.07 (m, 1H), 7.16-7.19 (m, 2H), 7.49-7.66 (m, 3H), 8.06
(d, 1H, J=8.3 Hz).
[0380] To a stirred solution of ethyl
(S)-3-methoxy-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-
-2-pyrrolidinyl]methoxy]benzoate (760 mg, 1.32 mmol) in THF (10 mL)
was added 0.25 N NaOH (10 mL), and the resulting mixture was heated
under reflux overnight. After cooling to room temp, the mixture was
poured into 1 N HCl (100 mL) and the solid was collected. The crude
solid was washed with Et.sub.2O to give 429 mg (59%) 27 as a yellow
amorphous solid. mp 132-135; IR (KBr) 1707 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.84-2.18 (m, 4H), 2.25 (s, 3H), 2.49-2.51
(m, 2H), 3.29-3.59 (m, 4H), 3.80 (s, 3H), 3.82 (s, 3H), 4.00-4.05
(m, 1H), 6.53-8.01 (m, 10H), 8.45 (s, 1H), 8.54 (s, 1H), 12.63 (bs,
1H); MS (FAB) m/z 548 (M.sup.++1).
Example 24
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nylmethoxy]phthalic acid
[0381] ##STR1649##
[0382] To a stirred solution of dimethyl 4-hydroxyphthalate (3.00
g, 14.27 mmol), N-Boc-prolinol (2.87 g, 14.26 mmol), Ph.sub.3P
(4.49 g, 17.12 mmol) in THF (50 mL) was added DIAD (3.40 mL, 17.27
mmol) at 0.degree. C. Then the resulting mixture was heated under
reflux overnight The resulting mixture was evaporated and the
residue was purified by column chromatography on silica-gel with
n-hexane-EtOAc (3:1, v/v) as eluent to give 5.75 g (q.y.) dimethyl
(S)-4-[1-tert-butoxycarbonyl)-2-pyrrolidinyl methoxy]phthalate as
an oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47 (s, 9H), 1.86-2.05
(m, 4H), 3.36-3.40 (m, 2H), 3.87 (m, 3H), 3.91 (s, 3H), 3.96-4.19
(m, 3H), 7.03-7.24 (m, 2H), 7.80 (m, 1H).
[0383] To a solution of dimethyl
(S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethoxy]phthalate
(5.75 g, 14.62 mmol) in CH.sub.2Cl.sub.2 (25 mL) was added TFA (20
mL), and the resulting mixture was stirred for 50 min at room temp.
The resulting mixture was concentrated in vacuo and made basic with
sat. NaHCO.sub.3. The mixture was extracted with CH.sub.2Cl.sub.2,
washed with brine, dried over MgSO.sub.4, and evaporated off in
vacuo. The residue was purified by column chromatography on
silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as eluent to give 790
mg (18%) dimethyl (S)-4-(2-pyrrolidinylmethoxy)phthalate as a brown
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.48-1.57 (m, 1H), 1.72-1.84
(m, 2H), 1.89-1.98 (m, 2H), 2.91-3.03 (m, 2H), 3.48-3.54 (m, 1H),
3.82-3.97 (m, total 8H), 6.98 (dd, 1H, J=2.4, 8.8 Hz), 7.06 (d, 1H,
J=2.4 Hz), 7.78 (d, 1H, J=8.8 Hz).
[0384] To a stirred solution of dimethyl
(S)-4-(2-pyrrolidinylmethoxy)phthalate (212 mg, 0.72 mmol) in DMF
(8 mL) was added pentafluorophenyl ester of the
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (346 mg,
0.72 mmol) and Et.sub.3N (120 ml, 0.86 mmol), and the mixture was
stirred overnight The resulting mixture was diluted with EtOAc,
washed with 1 N HCl, sat. NaHCO.sub.3, brine, and dried over
MgSO.sub.4. The solvent was evaporated off in vacuo to give 413 mg
(97%) dimethyl
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolid-
inylmethoxy]phthalate as an oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.92-2.12 (m, 4H), 2.29 (br s, 3H), 3.51-3.64 (m, 7H), 3.87 (s,
3H), 3.89 (s, 3H), 4.10-4.19 (m, 2H), 4.44 (m, 1H), 6.73-8.02
(series of m, total 12H).
[0385] To a stirred solution of dimethyl
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolid-
inylmethoxy]phthalate (413 mg, 0.70 mmol) in THF (10 mL) was added
0.25 N NaOH (10 mL) at room temp, and then the resulting mixture
was heated under reflux overnight. After cooling to room temp, the
reaction mixture was poured into 1 N HCl (100 mL). The solid was
collected, washed with water and air-red. The crude solid was
washed with Et.sub.2O to give 310 mg (79%) 28 as a yellow amorphous
solid. IR (KBr) 1701 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.87-2.18 (m, 4H), 2.25 (s, 3H), 2.50 (s, 2H), 3.38-3.60 (m, 4H),
3.83 (s, 3H), 4.00-4.14 (m, 1H), 6.74-8.02 (series of m, 10H), 8.46
(s, 1H), 8.54 (s, 1H); MS (FAB) m/z 562 (M.sup.+30 1).
Example 25
3-chloro-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyr-
rolidinyl]methoxy]benzoic acid
[0386] ##STR1650##
[0387] To a stirred solution of methyl 3-chloro-4-hydroxybenzoate
(600 mg, 3,215 mmol), N-tert-butoxycalbonylprolinol (647,1 mg,
3,215 mmol), and Ph.sub.3P (1.01 g, 3.858 mmol) in THF (10 mL) was
added dropwise diisopropyl azodicarboxylate (DIAD) (0.8 mL, 3.890
mmol) at room temp and the mixture was stirred for 3 days at room
temp, and for 18 hr at 70.degree. C. The reaction mixture was
evaporated off in vacuo, and the residue was chromatographed on
silica-gel with n-hexane:EtOAc (5:1, v/v) as eluent to give 1.147 g
(97%) methyl
3-chloro-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy benzoate
as an oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.46, 1.48 (s
each, 9H), 1.59-1.63 (br, 1H), 1.88 (br s, 1H), 2.05 (s, 1H),
2.05-2.21 (m, 2H), 3.34-3.45 (br m, 1.5H), 3.89 (s, 3H), 3.97 (br
m, 0.5H), 4.21 (br s, 2H), 7.05 (d, J=8.8 Hz, 1H), 7.90 (dd, J=2.0,
8.8 Hz, 1H), 8.04 (d, J=2.0 Hz, 1H); MS (FAB) m/z 370
(M.sup.++1).
[0388] To a stirred solution of methyl
3-chloro-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy benzoate
(1.14 g, 3.10 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added TFA (5
mL) at 0.degree. C., and the reaction mixture was stirred at room
temp for 2 hr. The solvent was removed under a reduced pressure and
the residue was treated with 1N NaOH, The mixture was extracted
with CHCl.sub.3. The extract was washed with brine, dried over KOH,
and concentrated under a reduced pressure to afford 741 mg (89%)
methyl 3-chloro-4-(2-pyrrolidinyl)methoxybenzoate as a yellow oil.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.60-1.67 (m, 1H),
1.78-2.02 (m, 3H), 2.93-2.98 (m, 1H), 3.03-3.09 (m, 1H), 3.59 (dt,
J=2.0, 9.3 Hz, 1H), 3.89 (s, 3H), 3.98 (dd, J=6.3, 8.8 Hz, 1H),
4.05 (dd, J=4.9, 9.3 Hz, 1H), 6.93 (d, J=8.8 Hz, 1H), 7.90 (dd,
J=2.0, 8.8 Hz, 1H), 8.04 (d, J=2.0 Hz, 1H); MS (FAB) m/z 270
(M.sup.++1).
[0389] The mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (500 mg, 1.04
mmol), methyl 3-chloro-4-(2-pyrrolidinyl)methoxybenzoate (281 mg,
1.04 mmol), Et.sub.3N (0.17 mL, 1.25 mmol) in DMF (5 mL) was
stirred for 1 hr at room temp. The mixture was diluted with EtOAc,
washed with brine, and dried over Na.sub.2SO.sub.4. The solvent was
removed under a reduced pressure, and the residue was
chromatographed on silica-gel with n-hexane-EtOAc (1:3, v/v) as
eluent to afford methyl
3-chloro-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-py-
rrolidinyl]methoxy]benzoate (620 mg, 1.04 mmol) as a white
crystalline solid. To a stirred solution of this compound in THF (8
mL) and H.sub.2O (2 mL) was added LiOH (74.9 mg, 3.126 mmol), and
the mixture was stirred at room temp for 2 days. The mixture was
diluted with CHCl.sub.3, and treated with 1N HCl. The solution was
washed with brine, dried over Na.sub.2SO.sub.4, and evaporated in
vacuo. The crude solid was recrystallized from
n-hexane-EtOAc--CHCl.sub.3 to afford 561.2 mg (98%) 29 as a white
crystalline material. IR (KBr) 1676, 1599, 1487, 1267, 758, 754
cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.82-2.24
(m, 4H), 2.25 (s, 3H), 3.48-3.60 (m, 4H), 3.78 (s, 3H), 4.18 (m,
2H), 4.31 (m, 1H), 6.74 (dd, J=1.5, 8.3 Hz, 1H), 6.84 (d, J=2.0 Hz,
1H), 6.91-6.95 (m, 1H), 7.11-7.17 (m, 3H), 7.79 (dd, J=2.0, 8.3 Hz,
2H), 7.85 (d, J=2.0 Hz, 1H), 7.98 (d, J=8.3 Hz, 1H), 8.53 (s, 1H),
8.58 (s, 1H); MS (FAB) m/z 552 (M.sup.++1).
Example 26
3,5-dichloro-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-
-pyrrolidinyl]methoxy]benzoic acid
[0390] ##STR1651##
[0391] To a stirred solution of methyl
3,5-dichloro-4-hydroxybenzoate (600 mg, 2.714 mmol),
N-tert-butoxycarbonylprolinol (546 mg, 2.714 mmol), and Ph.sub.3P
(854 mg, 3.257 mmol) in THF (10 mL) was added dropwise DIAD (0.68
mL, 3.283 mmol) at room temp and the mixture was stirred for 3 days
at room temp, and for 18 hr at 70.degree. C. The reaction mixture
was concentrated and the residue was chromatographed on silica-gel
with n-hexane-EtOAc (6:1, v/v) as eluent to give 988.8 mg (90%)
methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3,5-dichlorobenzoate
a pale yellowish oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.44 (s, 9H), 1.88-2.15 (br m, 3H), 2.34 (br s, 1H), 3.40-3.44 (m,
2H), 3.92 (s, 3H), 3.92, 4.14 (m, 1H), 4.18 (br s, 2H), 7.98 (s,
2H); MS (FAB) m/z 404 (M.sup.++1).
[0392] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3,5-dichlorobenzoate
(988 mg, 3.248 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added TFA (5
mL) at 0.degree. C., and the reaction mixture was stirred at room
temp for 2 hr. The solvent was removed under a reduced pressure and
the residue was treated with 1N NaOH, The solution was extracted
with CHCl.sub.3. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated under a reduced pressure to
afford 672 mg (68%) methyl
3,5-dichloro-4-(2-pyrrolidinyl)methoxybenzoate as a pale yellowish
oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.62-1.69 (m, 1H),
1.78-1.86 (m, 2H), 1.89-1.99 (m, 1H), 2.92-2.98 (m, 1H), 3.04-3.09
(m, 1H), 3.55-3.60 (m, 1H), 3.91 (s, 3H), 4.01 (dd, J=6.8, 8.8 Hz,
1H), 4.08 (dd, J=4.9, 8.8 Hz, 1H), 7.97 (s, 2H); MS (FAB) m/z 304
(M.sup.++1).
[0393] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (385.8 mg,
0.803 mmol), methyl 3,5-dichloro-4-(2-pyrrolidinyl)methoxybenzoate
(244.3 mg, 0.803 mmol), Et.sub.3N (0.13 mL, 0.964 mmol) in DMF (4
mL) was stirred for 1 hr at room temp. The mixture was diluted with
EtOAc, washed with brine, and dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure, and the residue was
chromatographed on silica-gel with n-hexane:EtOAc (1:2, v/v) as
eluent to afford methyl
3,5-dichloro-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]--
2-pyrrolidinyl]methoxy]benzoate as an oil. To a stirred solution of
this compound in THF (8 mL) and H.sub.2O (2 mL) was added LiOH
(57.7 mg, 2.409 mmol), and the mixture was stirred at room temp
overnight. The mixture was concentrated in vacuo, and the residue
was diluted with CHCl.sub.3. The solution was washed with 1N HCl,
brine, and dried over Na.sub.2SO.sub.4. The solvent was removed
under a reduced pressure, and the obtained crude solid was
recrystallized from n-hexane-MeOH--CHCl.sub.3 to afford 428.2 mg
(91%) 30 as a white crystalline powder. IR (KBr) 1618, 1535, 1454,
1257, 754 cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
1.83-2.24 (m, 4H), 2.24 (s, 3H), 3.50-3.58 (m, 4H), 3.84 (s, 3H),
3.98-4.05 (m, 1H), 4.15 (dd, J=2.9, 8.7 Hz, 1H), 4.29 (br m, 1H),
6.74 (d, J=8.3 Hz, 1H), 6.87 (s, 1H), 6.93 (t, J=7.3 Hz, 1H), 7.11
(d, J=7.8 Hz, 1H), 7.16 (d, J=8.3 Hz, 1H), 7.79 (d, J=8.3 Hz, 1H),
7.86 (s, 1H), 7.87 (d, J=9.8 Hz, 1H), 7.99 (d, J=8.3 Hz, 1H), 8.49
(s, 1H), 8.58 (s, 1H); MS (FAB) m/z 586 (M.sup.++1)
Example 27
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinylm-
ethoxy]-3-nitrobenzoic acid
[0394] ##STR1652##
[0395] To a stirred solution of 4-hydroxy-3-nitrobenzoic acid (3.00
g, 0.0164 mol) in MeOH-benzene (1:4, v/v) was added dropwise 2.0
M-n-hexane solution of TMSCHN.sub.2 (8.2 mL, 0.0164 mol) at room
temp. After the resulting solution was stirred for 4 hr at room
temp, the mixture was evaporated off in vacuo. The oily residue was
chromatographed on silica-gel with CHCl.sub.3 as eluent to afford
4.23 g (79%) methyl 4-hydroxy-3-nitrobenzoate as a pale yellow
crystalline material.
[0396] To a stirred mixture of N-tert-butoxycarbonylprolinol (1.02
g, 5.07 mmol), methyl 4-hydroxy-3-nitrobenzoate (1.0 g, 5.07 mmol),
and Ph.sub.3P (1.46 g, 5.58 mmol) in THF (10 mL) was added dropwise
diisopropyl azodicarboxylate (DIAD) (95%) (1.16 mL, 5.58 mmol) at
0.degree. C. The resulting mixture was heated under reflux for 46
hr. After cooling, the mixture was evaporated off in vacuo. The
residue was dissolved in CH.sub.2Cl.sub.2 (10 mL) and added TFA
(100 mL). After the solution was stirred for 0.5 hr at room temp,
the solution was evaporated in vacuo. Water was added to the
residue and washed with EtOAc. The aqueous layer was neutralized by
the addition of sat. NaHCO.sub.3 and extracted with EtOAc. The
extract was dried over Na.sub.2SO.sub.4 and evaporated in vacuo to
afford 0.698 g (49%) methyl
3-nitro-4-(2-pyrrolidinylmethoxy)benzoate as a gum.
[0397] A mixture of methyl
3-nitro-4-(2-pyrrolidinylmethoxy)benzoate (0.668 g, 2.38 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.12 g,
3.57 mmol), 1-hydroxybenzo triazole (HOBt) (0.482 g, 3.57 mmol),
4-dimethylaminopyridine (DMAP) (43.6 mg, 0.357 mmol), and
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (0.684 g, 3.57
mmol) in DMF (10 mL) was stirred for 15 hr at room temp. EtOAc was
added to the mixture and washed successively with 1 N HCl, sat.
NaHCO.sub.3, and brine. The organic layer was dried over
Na.sub.2SO.sub.4 and evaporated in vacuo. The residue was
chromatographed on silica-gel with EtOH--CHCl.sub.3 (1:20, v/v) as
eluent to afford 0.927 g (68%) methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacethyl]-2-pyrrolidiny-
lmethoxy]-3-nitrobenzoate as a yellow crystalline material.
[0398] A mixture of methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacethyl]-2-pyrrolidiny-
lmethoxy]-3-nitrobenzoate (0.917 g, 1.59 mmol) in THF (10 mL) and 1
N NaOH (2.38 mL, 2.38 mmol) was heated under reflux for 2 hr. After
cooling, the mixture was poured into ice water and extracted with
EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated in vacuo to afford 0.826 g (92%) 31
as a yellow crystalline solid. .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. 1.91, 2.09 (1H, 3H, each m), 2.28 (3H, s), 3.54-3.62 (4H,
m), 3.64 (3H, s), 4.15, 4.59 (each 1H, each d, J=7.8 Hz), 4.46 (1H,
m), 6.66, 7.22 (each 1H, each s), 6.72 (1H, d, J=8.3 Hz), 7.11-7.28
(4H, m), 7.46 (1H, d, J=7.8 Hz), 7.74 (1H, d, J=7.8 Hz), 7.85 (1H,
s), 8.17 (1H, dd, J=2.0, 8.8 Hz), 8.48 (1H, d, J=2.4 Hz); MS (FAB)
m/z 563 (M.sup.++1).
Example 28
3-amino-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrro-
lidinylmethoxy]benzoic acid
[0399] ##STR1653##
[0400] A stirred mixture of
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacethyl]-2-pyrrolidiny-
lmethoxy]-3-nitrobenzoic acid (101 mg, 0.190 mmol) and 5% Pd--C
(0.247 g) in methanol was hydrogenated at 1 atm for 48 hr.
Insoluble catalyst was removed, and the filtrate was evaporated in
vacuo. The residue was chromatographed on silica-gel with
EtOH--CHCl.sub.3 (1:1, v/v) as eluent to afford 61.0 mg (60%) 32 as
a crystalline material. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
1.95 (4H, m), 2.23 (3H, s), 3.60, 3.91, 4.10, 4.34 (5H, each m),
3.81 (3H, s), 4.88 (2H, m), 6.74 (1H, d, J=8.3 Hz), 6.86-7.28 (5H,
m), 7.78 (1H, d, J=7.8 Hz), 7.99 (1H, d, J=8.3 Hz), 8.30 (1H, s),
8.45, 8.55 (each 1H, each s); MS (FAB) m/z 533 (M.sup.++1).
Example 29
4-[2-[1-[4-[N'-(2-fluorophenyl)ureido)-3-methoxyphenylacetyl]-2-pyrrolidin-
yl]ethynyl]benzoic acid
[0401] ##STR1654##
[0402] To a stirred solution of benzyl
4-amino-3-methoxyphenylacetate (1.36 g, 5 mmol) in THF (20 mL) was
added 2-fluorophenyl isocyanate (561 ul, 5 mmol) and a catalytic
amount of Et.sub.3N. The resulting mixture was stirred for 3 hr.
The mixture was quenched by the addition of H.sub.2O (10 mL) and
extracted with EtOAc. The extract was washed with brine, dried over
MgSO.sub.4, and evaporated. The residue was chromatographed on
silica-gel with CHCl.sub.3 as eluent to give 2.06 g (q.y.) benzyl
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetate as a green
oil. .sup.1H-NMR (CDCl).sub.3 .delta. 3.63 (2H, s), 3.82 (3H, s),
5.14 (2H, s), 6.79-7.37 (12H, m), 8.01 (1H, d, J=7.8 Hz), 8.09-8.14
(1H, m).
[0403] To a stirred solution of benzyl
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetate (2.04 g, 5
mmol) in THF (40 mL) was added 0.25 N NaOH (40 mL). The resulting
mixture was stirred overnight. The mixture was poured into 1 N HCl
(10 mL), and the resulting precipitate was collected with suction.
The residue was recrystallized from CHCl.sub.3-EtOH to give 1.04 g
(66%) 4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetic acid as a
white crystalline powder. mp 185-188 (d); .sup.1H-NMR
(DMSO-d.sub.6) .delta. 3.50 (2H, s), 3.82 (3H, s), 6.78 (1H, dd,
J=1.4 and 8.3 Hz), 6.92 (1H, d, J=1.4 Hz), 6.95-7.01 (1H, m),
7.10-7.14 (1H, m), 7.19-7.24 (1H, m), 8.01 (1H, d, J=8.3 Hz),
8.14-8.18 (1H, m), 8.72 (1H, s), 9.17 (1H, s); MS (FAB) m/z 319
(M.sup.++1); Anal. Calcd for C.sub.16H.sub.15N.sub.2O.sub.4F: C,
60.37; H, 4.75; N, 8.80. Found: C, 60.20; H, 4.82; N, 8.67.
[0404] A mixture of
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetic acid (255 mg,
0.8 mmol), 2-[2-(4-ethoxycarbonylphenyl)ethynyl]pyrrolidine (195
mg, 0.8 mmol), EDC (230 mg, 1.2 mmol), DMAP (98 mg, 0.8 mmol) in
DMF (20 mL) was stirred overnight. The reaction mixture was poured
into 1 N HCl and the resulting precipitate was collected with
suction and dissolve in CHCl.sub.3. The solution was dried over
MgSO.sub.4 and evaporated. The residue was chromatographed on
silica gel with CHCl.sub.3-MeOH (100:1, v/v) as eluent to give the
desired compound, which was dissolved in THF (8 mL). 0.25 N NaOH (8
mL) was added to this solution and the resulting mixture was
stirred overnight. The mixture was poured into 1 N HCl and
extracted with CHCl.sub.3. The extract was washed with brine, dried
over MgSO.sub.4, and evaporated. The residue was recrystallized
from CHCl.sub.3-n-hexane to give 144 mg (37%) 33 as a pale yellow
crystalline powder. mp 152-155 (d); .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.92-2.27 (4H, m), 2.50 (2H, s), 3.33-3.78 (2H, m), 3.80
and 3.82 (total 3H, s, each), 4.88-5.12 (1H, m), 6.77-7.24 and
7.99-8.20 (total 7H, m), 7.48 and 7.52 (2H, d, J=8.3 Hz, each),
7.91 (2H, d, J=8.3 Hz), 8.72 (1H, s), 9.18 (1H, s), 13.11 (1H,
br-s); MS (FAB) m/z 516 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.26N.sub.3O.sub.3F.2H.sub.2O: C, 63.15; H, 5.48; N,
7.62. Found: C, 63.58; H, 5.15; N, 7.22.
Example 30
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methoxy]-3-methylbenzoic acid
[0405] ##STR1655##
[0406] To a stirred solution of 4-iodo-2-methylphenol (465 mg,
1.987 mmol), N-tert-butoxycarbonylprolinol (400 mg, 1.987 mmol),
and Ph.sub.3P (625 mg, 2.384 mmol) in THF (7 mL) was added dropwise
DIAD (0.5 mL, 2.404 mmol) at room temp, and the mixture was stirred
for 13 hr at 70.degree. C. The reaction mixture was concentrated in
vacuo and the residue was chromatographed on silica-gel with
n-hexane-EtOAc (9:1, v/v) as eluent to give 645.3 mg (78%)
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-1-iodo-3-methylbenzene
as a pale yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.47 (s, 9H), 1.83-1.89 (m, 1H), 1.96-2.04 (m, 3H), 2.16 (s, 3H),
3.37-3.43 (br m, 2H), 3.81, 3.94 (br m each, 1H), 4.08-4.18 (m,
2H), 6.62 (br s, 1H), 7.42 (s, 2H); MS (FAB) m/z 418
(M.sup.++1).
[0407] To a stirred solution of
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-1-iodo-3-methylbenzene
(645.3 mg, 1.546 mmol) in DMSO (7 mL) and MeOH (6 mL) was added
Et.sub.3N (0.47 mL, 3.401 mmol), Pd(OAc).sub.2 (17.4 mg, 0.077
mmol) and 1,3-bis(diphenylphosphino) propane (31.46 mg, 0.077
mmol). To the stirred mixture was induced CO gas for 10 min. The
mixture was stirred at 70.degree. C. for 2 days and concentrated.
The residue was diluted with EtOAc, washed with brine, and dried
over Na.sub.2SO.sub.4. The solvent was removed under a reduced
pressure, and the residue was chromatographed on silica-gel with
n-hexane-EtOAc (5:1, v/v) as eluent to afford 301.6 mg (56%)
methyl-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-methylbe-
nzoate as an oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.47
(s, 9H), 1.86-2.10 (br m, 4H), 2.33 (s, 3H), 3.32-3.50 (br m, 2H),
3.88 (s, 3H), 3.88, 4.04 (br m each, 1H), 4.13-4.20 (m, 2H), 6.89
(br m, 1H), 7.82 (s, 1H), 7.85 (dd, J=2.0, 8.8 Hz, 1H); MS (FAB)
m/z 350 (M.sup.++1).
[0408] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methyloxy-3-methylbenzoate
(301.6 mg, 0.863 mmol) in CH.sub.2Cl.sub.2 (6 mL) was added TFA
(1.2 mL) at 0.degree. C., and the mixture was stirred at room temp
for 1 hr. The solvent was removed under a reduced pressure, and the
residue was made basic by the addition of 1N NaOH, The mixture was
extracted with CHCl.sub.3. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated under a reduced pressure to
afford 192.5 mg (90%) methyl
3-methyl-4-(2-pyrrolidinyl)methoxybenzoate as an oil. .sup.1H-NMR
(400 MHz, CDCl.sub.3) .delta. 1.58-1.65 (m, 1H), 1.78-2.00 (m, 3H),
2.24 (s, 3H), 2.97 (dt, J=6.8, 10.2 Hz, 1H), 3.05 (dt, J=5.9, 6.8
Hz, 1H), 3.54-3.58 (m, 1H), 3.87 (s, 3H), 3.92 (dd, J=6.3, 9.3 Hz,
1H), 3.99 (dd, J=4.9, 9.3 Hz, 1H), 6.81 (d, J=8.3 Hz, 1H), 7.83 (s,
1H), 7.85 (dd, J=2.0, 8.3 Hz, 1H); MS (FAB) m/z 250
(M.sup.++1).
[0409] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (211.3 mg,
0.44 mmol), methyl 3-methyl-4-(2-pyrrolidinyl)methoxybenzoate
(109.7 mg, 0.44 mmol), Et.sub.3N (73.6 ul, 0.528 mmol) in DMF (2
mL) was stirred for 1.5 hr at room temp. The reaction mixture was
diluted with EtOAc. The solution was washed with brine and dried
over Na.sub.2SO.sub.4. The solvent was removed under a reduced
pressure, and the residue was chromatographed on silica-gel with
n-hexane-EtOAc (1:3, v/v) as eluent to afford methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methoxy]-3-methyl benzoate (241.6 mg, q.y.) as an oil. To a
stirred solution of this compound in THF (4.4 mL) and H.sub.2O (1.1
mL) was added LiOH (32 mg, 1.32 mmol), and the reaction mixture was
stirred at room temp overnight. The mixture was diluted with
CHCl.sub.3, and acidified by the addition of 1N HCl. The solution
was washed with brine and dried over Na.sub.2SO.sub.4. The solvent
was removed under a reduced pressure, and the obtained crude solid
was recrystallized from n-hexane-EtOAc--CHCl.sub.3-MeOH to afford
126.3 mg (54%) 34 as a white crystalline powder. IR (KBr) 1685,
1606, 1454, 1257, 752 cm.sup.-1; .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.87-2.10 (m, 4H), 2.12 (s, 3H), 2.25 (s,
3H), 3.51-3.71 (m, 4H), 3.76 (s, 3H), 4.08-4.18 (m, 2H), 4.34 (m,
1H), 6.74 (dd, J=1.5, 9.8 Hz, 1H), 6.84 (d, J=1.5 Hz, 1H), 6.94 (t,
J=6.8 Hz, 1H), 7.06 (d, J=8.8 Hz, 1H), 7.12 (d, J=7.8 Hz, 1H), 7.16
(d, J=7.8 Hz, 1H), 7.72 (s, 1H), 7.76 (dd, J=2.0, 8.3 Hz, 1H), 7.79
(d, J=7.8 Hz, 1H), 7.99 (d, J=8.3 Hz, 1H), 8.46 (s, 1H), 8.54 (s,
1H); MS (FAB) m/z 532 (M.sup.++1); Anal. Calcd for
C.sub.30H.sub.33N.sub.3O.sub.6.1/2H.sub.2O: C, 66.65; H, 6.34; N,
7.77. Found: C, 66.16; H, 6.37; N, 7.50.
Example 31
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nylmethoxy]isophthalic acid
[0410] ##STR1656##
[0411] To a solution of dimethyl 4-acetoxyisophthalate (1.52 g,
6.03 mmol) in MeOH (70 mL) was added sat. NaHCO.sub.3, and the
resulting mixture was stirred for 3 hr at room temp. The resulting
mixture was poured into 1 N HCl and extracted with EtOAc. The
extract was washed with sat NaHCO.sub.3, brine, and dried over
MgSO.sub.4. The solvent was evaporated to give 1.27 g (q.y.)
dimethyl 4-hydroxy isophthalate as a white crystalline powder.
.sup.1H-NMR (CDCl.sub.3) .delta. 3.91 (s, 3H), 3.99 (s, 3H), 7.01
(d, 1H, J=8.8 Hz), 8.11 (dd, 1H, J=2.4, 8.8 Hz), 8.55 (d, 1H, J=2.4
Hz).
[0412] To a stirred solution of dimethyl 4-hydroxyisophthalate
(1.27 g, 6.04 mmol), (S)--N-Boc-Prolinol (1.22 g, 6.06 mmol),
PPh.sub.3 (1.90 g, 7.24 mmol) in THF (30 mL) was added DIAD (1.43
mL, 7.26 mmol) at room temp. The resulting stirred mixture was then
heated under reflux for 15 hr. After cooling to room temp, the
resulting mixture was evaporated and the residue was purified by
column chromatography on silica-gel with n-hexane-EtOAc (3:1, v/v)
as eluent to give 2.10 g (88%) dimethyl
(S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethoxy]isophthalate
as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.26 (s, 9H),
1.85-2.16 (m, 3H), 3.36-3.46 (m, 2H), 3.90 (s, 6H), 4.11-4.31 (m,
2H), 4.95-5.02 (m, 2H), 7.09 (dd, 1H, J=9.3, 24.9 Hz), 8.11-8.14
(m, 1H), 8.46 (d, 1H, J=9.3 Hz)
[0413] A mixture of dimethyl
(S)-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethoxy]isophthalate
(2.01 g, 5.11 mmol), TFA (20 mL), and CH.sub.2Cl.sub.2 (25 mL) was
stirred for 1.5 hr at room temp. The resulting mixture was
concentrated in vacuo and made basic with sat. NaHCO.sub.3. The
mixture was extracted with CH.sub.2Cl.sub.2, washed with brine,
dried over Na.sub.2CO.sub.3, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (9:1, v/v) as eluent to give 0.80 g (53%) dimethyl
(S)-4-(2-pyrrolidinylmethoxy)isophthalate as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.71 (m, 1H), 1.89 (m, 2H), 2.00
(m, 1H), 3.05-3.13 (m, 2H), 3.67 (m, 1H), 3.90 (s, 3H), 3.91 (s,
3H), 4.05-4.18 (m, 2H), 7.00 (d, 1H, J=8.8 Hz), 8.14 (dd, 1H,
J=2.4, 8.8 Hz), 8.50 (d, 1H, J=2.4 Hz).
[0414] To a stirred solution of dimethyl
(S)-4-(2-pyrrolidinylmethoxy)isophthalate (616 mg, 2.10 mmol) in
DMF (13 mL) was added pentafluoro ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.00 g,
2.08 mmol) and Et.sub.3N (425 .mu.l, 3.12 mmol), and the resulting
mixture was stirred for 3.5 hr at room temp. The resulting mixture
was diluted with EtOAc, washed with 1 N HCl, sat. NaHCO.sub.3,
brine, and dried over Na.sub.2SO.sub.4. After removal of the
solvent, the residue was purified by column chromatography on
silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as eluent to give 1.41
g (q.y.) dimethyl
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolid-
inylmethoxy]isophthalate as a yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.86-2.29 (m, 4H), 2.30 (s, 3H), 3.47-3.57 (m, 2H), 3.58
(s, 3H), 3.59 (s, 2H), 3.83 (s, 3H), 3.91 (s, 3H), 4.22-4.37 (m,
2H), 4.42-4.47 (m, 1H), 6.44-8.46 (series of m, 12H).
[0415] To a stirred solution of dimethyl
(S)-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinylmethoxy]isophthalate (1.41 g, 2.39 mmol) in
THF (20 mL) was added 0.25 N NaOH (20 mL), and the resulting
mixture was then heated under reflux overnight. After cooling to
room temp, the mixture was poured into 1 N HCl (150 mL) and the
solid was collected. The crude solid was recrystallized from
CHCl.sub.3-MeOH to give 140 mg (10%) 35 as a white crystalline
powder. .sup.1H-NMR (CDCl.sub.3) .delta. 1.83-2.18 (m, 4H), 2.24
(s, 3H), 3.44-3.55 (m, 4H), 3.59 (s, 2H), 3.80 (s, 3H), 4.05-4.24
(m, 2H), 4.28-4.32 (m, 1H), 6.73-8.55 (series of m, total 12H); MS
(FAB) m/z 562 (M.sup.++1); Anal. Calcd. for
C.sub.30H.sub.31N.sub.3O.sub.8.4H.sub.2O: C, 56.87; H, 6.20; N,
6.63. Found: C, 56.73; H, 5.56; N, 6.52.
Example 32
3-methoxy-4-[2-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2--
pyrrolidinyl]ethynyl]benzoic acid
[0416] ##STR1657##
[0417] A stirred mixture of methyl 3-methoxy-4-nitrobenzoate (1.20
g, 5.7 mmol) and 5% Pd--C (1.0 g) in EtOH (30 mL) and THF (20 mL)
was hydrogenated overnight at 1 atm. The mixture was filtered and
the filtrate was evaporated. The residue was chromatographed on
silica-gel with CHCl.sub.3 as eluent, and the solid obtained was
further purified by recrystallization from CHCl.sub.3-n-hexane to
give 805 mg (78%) methyl 4-amino-3-methoxybenzoate as white plates.
mp 126-128; IR (KBr) 3475, 1700 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3)
.delta. 3.86 (3H, s), 3.89 (3H, s), 4.21 (2H, br s), 6.66 (1H, d,
J=8.3 Hz), 7.45 (1H, d, J=1.9 Hz), 7.54 (1H, dd, J=1.9 and 8.3 Hz);
MS (FAB) m/z 182 (M.sup.++1); Anal. Calcd for
C.sub.9H.sub.11NO.sub.3: C, 59.66; H, 6.12; N, 7.73. Found: C,
59.65; H, 6.15; N, 7.65.
[0418] A stirred solution of methyl 4-amino-3-methoxybenzoate (725
mg, 4 mmol) in EtOH (10 mL) was added to dil.H.sub.2SO.sub.4
(prepared from H.sub.2SO.sub.4 0.5 mL and H.sub.2O 10 mL) at
0.degree. C. A solution of NaNO.sub.2 (331 mg, 4.8 mmol) in
H.sub.2O (10 mL) was added to the mixture. After stirring for 0.5
hr at the same temp, the mixture was poured into a cooled
(0.degree. C.), stirred suspended solution of KI (1.83 g, 11 mmol)
and cat. Cu in H.sub.2O (100 mL). The mixture was vigorously
stirred for 1 hr at room temp and extracted with CHCl.sub.3. The
extract was washed with brine, dried over MgSO.sub.4, and
evaporated. The residue was chromatographed on silica-gel with
n-hexane-EtOAc (10:1, v/v) as eluent to give a mixture of methyl
4-iodo-3-methoxybenzoate and methyl 3-methoxybenzoate (748 mg) as a
colorless oil.
[0419] To this oil was added Pd(PPh.sub.3).sub.4 (150 mg, 0.13
mmol), CuI (57 mg, 0.3 mmol) and i-Pr.sub.2NH (10 mL). The mixture
was stirred for 1 hr under N.sub.2 and a solution of
1-(tert-butoxycarbonyl)-2-ethynylpyrrolidine (488 mg, 2.5 mmol) in
i-Pr.sub.2NH (10 mL) was added to the mixture. After stirring for 2
hr, the mixture was poured into H.sub.2O and extracted with EtOAc.
The extract was washed with brine, dried over MgSO.sub.4, and
evaporated. The residue was chromatographed on silica-gel with
n-hexane-EtOAc (5:1, v/v) as eluent to give 431 mg (48%)
1-(tert-butoxycarbonyl)-2-[2-(2-methoxy-4-methoxycarbonylphenyl)ethynyl]p-
yrrolidine as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.49
(9H, s), 1.77-2.14 (4H, m), 3.36-3.51 (2H, m), 3.90 (3H, s), 3.91
(3H, s), 4.60-4.81 (1H, m), 7.36-7.39 (1H, m), 7.51 (1H, s),
7.55-7.57 (1H, m).
[0420] To a stirred solution of
1-(tert-butoxycarbonyl)-2-[2-(2-methoxy-4-methoxycarbonyl
phenyl)ethynyl]pyrrolidine (395 mg, 1.1 mmol) in CH.sub.2Cl.sub.2
(3 mL) was added TFA (3 mL). The resulting mixture was stirred for
1 hr. The mixture was concentrated in vacuo and made basic by the
addition of sat. NaHCO.sub.3, and extracted with CHCl.sub.3. The
extract was washed with H.sub.2O, dried over MgSO.sub.4, and
evaporated to give 238 mg (84%)
2-[2-(2-methoxy-4-methoxycarbonylphenyl)ethynyl]pyrrolidine as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.81-2.16 (4H, m),
2.97-3.17 (2H, m), 3.91 (6H, s), 4.13-4.15 (1H, m), 7.41 (1H, d,
J=8.3 Hz), 7.51 (1H, s), 7.56 (1H, d, J=8.3 Hz).
[0421] A mixture of
2-[2-(2-methoxy-4-methoxycarbonylphenyl)ethynyl]pyrrolidine (233
mg, 0.9 mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (314 mg, 1 mmol), EDC (268 mg, 1.4 mmol), DMAP (110 mg, 0.9
mmol) in DMF (10 mL) was stirred overnight. The mixture was poured
into 1 N HCl and the resulting solid was collected with suction.
The solid obtained was dissolved in CHCl.sub.3, and the solution
was dried over MgSO.sub.4 and evaporated. The residue was subjected
to short column chromatography on silica-gel with EtOAc as eluent
to give an oil. The oil was dissolved in THF (5 mL) and 0.25 N NaOH
was added to this solution with stirring. The solution was poured
into ice-1 N HCl to give a solid. The solid was collected, washed
with water, and air-dried. The crude solid was recrystallized from
CHCl.sub.3-n-hexane to give 215 mg (44%) 36 as a white crystalline
powder. mp 141-145; IR (KBr) 3338, 2956, 2935, 2875, 2593, 1711
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.91-2.14 (4H, m),
2.24 (3H, s), 3.38-3.68 (4H, m), 4.88-5.08 (1H, m), 6.76-8.56 (12H,
m); MS (FAB) m/z 542 (M.sup.++1).
Example 33
3-N,N-dimethylamino-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylace-
tyl]-2-pyrrolidinylmethoxy]benzoic acid
[0422] ##STR1658##
[0423] A stirred mixture of methyl 4-hydroxy-3-nitrobenzoate (3.22
g, 0.0163 mol) and 5% Pd--C (12.9 g) in MeOH (30 mL) was
hydrogenated under an atmospheric pressure for 70 hr at room temp.
Insoluble catalyst was removed, and the filtrate was evaporated in
vacuo. The residue was chromatographed on silica-gel with
EtOH--CHCl.sub.3 (1:20, v/v) as eluent to afford 1.89 g (69%)
methyl 3-amino-4-hydroxybenzoate as a pale brown syrup.
[0424] A stirred mixture of methyl 3-amino-4-hydroxybenzoate (1.07
g, 6.40 mmol) and 5% Pd--C (2.14 g) in MeOH (20 mL) and 37% aq.
formaldehyde (1.08 mL, 0.0122 mol) and 1 N HCl (6.1 mL) was
hydrogenated under an atmospheric pressure for 26 hr at room temp.
Insoluble catalyst was removed, and the filtrate was evaporated in
vacuo. The residue was chromatographed on silica-gel with
EtOAc-n-hexane (1:10, v/v) as eluent to afford 0.817 g (70%) methyl
3-(N,N-dimethylamino)-4-hydroxybenzoate as a syrup.
[0425] To a stirred mixture of methyl
3-(N,N-dimethylamino)-4-hydroxybenzoate (0.817 g, 4.18 mmol),
N-tert-butoxycarbonylprolinol (0.926 g, 4.60 mmol), Ph.sub.3P (1.21
g, 4.60 mmol) in THF (20 mL) was added dropwise DIAD (95%) (0.953
mL, 4.60 mmol) at 0.degree. C. The resulting mixture was heated
under reflux for 41 hr. After cooling, the mixture was evaporated
off in vacuo. The residue was chromatographed on silica-gel with
EtOAc-n-hexane (1:10 to 1:6, v/v) as eluent to give a syrup which
was used for the subsequent reaction without further purification.
This syrup was dissolved in CH.sub.2Cl.sub.2 (10 mL) and added TFA
(10 mL). After the solution was stirred for 5 hr at room temp, the
solution was evaporated in vacuo. Water was to the residue and
washed with CHCl.sub.3. The aqueous layer was neutralized by the
addition of sat. NaHCO.sub.3 and extracted with CHCl.sub.3. The
extract was dried over Na.sub.2SO.sub.4 and evaporated in vacuo to
afford 1.03 g (89%) methyl
3-(N,N-dimethylamino)-4-(2-pyrrolidinylmethoxy)benzoate as a
gum.
[0426] A mixture of methyl
3-(N,N-dimethylamino)-4-(2-pyrrolidinylmethoxy)benzoate (0.529 g,
1.90 mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (0.597 g, 1.90 mmol), HOBt (0.308 g, 2.28 mmol),
4-dimethylaminopyridine (DMAP) (23.2 mg, 0.190 mmol), and
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (0.437 g, 2.28
mmol) in DMF (10 mL) was stirred for 15 hr at room temp. The
mixture was neutralized by the carefully addition of 1 N HCl and
extracted with EtOAc. The extract was dried over Na.sub.2SO.sub.4
and evaporated in vacuo to afford 0.607 g (56%) methyl
3-N,N-dimethylamino-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]-phenyla-
cetyl]-2-pyrrolidinyl methoxy]benzoate as a white crystalline
material.
[0427] A mixture of methyl
3-(N,N-dimethylamino-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)
ureido]-phenylacetyl]-2-pyrrolidinylmethoxy]benzoate (0.600 g, 1.04
mmol) in THF (10 mL) and 0.25 N NaOH (5 mL, 1.25 mmol) was stirred
for 21 hr at room temp. CHCl.sub.3 was added to the mixture and
extracted with a mixture of water (100 mL)-1N NaOH (4 mL). The
extract was neutralized with sat. NH.sub.4Cl and extracted with
CHCl.sub.3. The extract was dried over Na.sub.2SO.sub.4 and
evaporated in vacuo to afford 428 mg (70%) 37 as a white
crystalline solid. .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
1.88, 1.99 and 2.11 (4H, each m), 2.24 (3H, s), 2.67 (6H, s), 3.33
(2H, m), 3.58 (2H, m), 4.05-4.32 (3H, m), 6.75 (1H, d, J=7.3 Hz),
6.92-6.95 (1H, m), 7.05 (1H, d, J=8.3 Hz), 7.11-7.17 (2H, m), 7.42
(1H, s), 7.52 (1H, d, J=7.8 Hz), 7.79 (1H, d, J=7.8 Hz), 8.00 (1H,
d, J=7.8 Hz), 8.31 (1H, s), 8.46, 8.55 (each 1H, each s); MS (FAB)
m/z 533 (M.sup.++1).
Example 34
3-fluoro-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyr-
rolidinyl]methoxy]benzoic acid
[0428] ##STR1659##
[0429] To a stirred solution of 4-bromo-2-fluorophenol (217 ul,
2.002 mmol), N-tert-butoxycarbonyl prolinol (403 mg, 2.002 mmol),
and Ph.sub.3P (630 mg, 2.403 mmol) in THF (7 mL) was added DIAD
(477 ul, 2.423 mmol) at room temp. The resulting mixture was
stirred for 6 hr at room temp and then overnight at 70.degree. C.
The mixture was concentrated in vacuo and the residue was
chromatographed on silica-gel with n-hexane-EtOAc (5:1, v/v) as
eluent to give 549.4 mg (73%)
1-bromo-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-fluorobenzene
as an oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.46 (s, 9H),
1.85 (br m, 1H), 1.90-2.10 (br s, 3H), 3.30-3.47 (m, 2H), 3.85,
4.04 (br s each, 1H), 4.11-4.20 (m, 2H), 6.82-6.98 (m, 1H),
7.13-7.26 (m, 2H); MS (FAB) m/z 374 (M.sup.++1).
[0430] To a stirred solution of
1-bromo-4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-fluorobenzene
(549.4 mg, 1.468 mmol) in DMSO (6 mL) and MeOH (5 mL) was added
Et.sub.3N (448 ul, 3.229 mmol), Pd(OAc).sub.2 (36.2 mg, 0.161
mmol), and 1,3-bis(diphenylphosphino)propane (66.4 mg, 0.161 mmol).
To the mixture was induced CO gas for 10 min. The resulting mixture
was stirred at 70.degree. C. for 2 days under a current of CO.
After the mixture was concentrated, the residue was diluted with
EtOAc. The solution was washed with brine and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure
and the residue was chromatographed on silica-gel eluting with
n-hexane:EtOAc (5:1, v/v) as eluent to afford 323.0 mg (62%) methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-fluorobenzoate
as a pale yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.47 (s, 9H), 1.87 (br s, 1H), 1.95-2.10 (m, 3H), 3.34-3.44 (br m,
2H), 3.89 (s, 3H), 3.94 and 4.11-4.26 (br m each, 3H), 7.03-7.11
(m, 1H), 7.75-7.80 (m, 2H); MS (FAB) m/z 354 (M.sup.++1).
[0431] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-fluorobenzoate
(323.0 mg, 0.914 mmol) in CH.sub.2Cl.sub.2 (6.5 mL) was added TFA
(1.3 mL) at 0.degree. C., and the mixture was stirred 1.5 hr at
room temp. The solvent was removed under a reduced pressure and the
residue was made basic by the addition of 1N NaOH, The mixture was
extracted with CHCl.sub.3. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated under a reduced pressure to
afford 174.8 mg (76%) methyl
3-fluoro-4-(2-pyrrolidinyl)methoxybenzoate as a brownish oil.
.sup.1H-NMR (400, CDCl.sub.3) .delta. 1.54-1.63 (m, 1H), 1.76-2.02
(m, 3H), 2.93-3.07 (m, 2H), 3.57 (ddd, J=4.9, 6.9, 14.3 Hz, 1H),
3.89 (s, 3H), 3.97 (dd, J=6.8, 9.3 Hz, 1H), 4.04 (dd, J=5.0, 8.8
Hz, 1H), 6.98 (t, J=17.6 Hz, 1H), 7.73 (dd, J=2.0, 11.7 Hz, 1H),
7.78 (dt, J=2.0, 8.8 Hz, 1H); MS (FAB) m/z 253 (M.sup.++1).
[0432] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (324.5 mg,
0.676 mmol), methyl 3-fluoro-4-(2-pyrrolidinyl)methoxybenzoate
(171.1 mg, 0.676 mmol), Et.sub.3N (113 ul, 0.811 mmol) in DMF (5
mL) was stirred for 2 hr at room temp. The mixture was diluted with
EtOAc, washed with brine, and dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure and the residue was
chromatographed on silica-gel with n-hexane:EtOAc (1:2, v/v) as
eluent to afford methyl
3-fluoro-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-py-
rrolidinyl]methoxy]benzoate (365.1 mg, 0.664 mmol) as an oil. To a
stirred solution of this compound in THF (4.4 mL) and H.sub.2O (1.1
mL) was added LiOH (46.3 mg, 1.932 mmol), and the reaction mixture
was stirred at room temp overnight. The mixture was diluted with
CHCl.sub.3 and acidified by the addition of 1N HCl. The separated
organic layer was washed with brine and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure,
and the obtained crude solid was recrystallized from
n-hexane-EtOAc--CHCl.sub.3 to afford 102 mg (30%) 38 as a white
crystalline powder. mp 123-126; IR (KBr) 1616, 1537, 1282, 756
cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.87-2.09
(m, 4H), 2.25 (s, 3H), 3.48-3.57 (m, 2H), 3.60 (s, 2H), 3.83 (s,
3H), 4.11-4.16 (m, 1H), 4.24 (dd, J=2.9, 9.8 Hz, 1H), 4.28-4.34 (br
s, 1H), 6.74 (dd, J=1.5, 8.3 Hz, 1H), 6.87 (s, 1H), 6.94 (t, J=7.3
Hz, 1H), 7.12 (d, J=7.8 Hz, 1H), 7.15 (t, J=8.3 Hz, 1H), 7.34 (t,
J=8.8 Hz, 1H), 7.66 (dd, J=2.0, 12.2 Hz, 1H), 7.73 (d, J=9.3 Hz,
1H), 7.79 (d, J=8.3 Hz, 1H), 7.99 (d, J=7.8 Hz, 1H), 8.46 (s, 1H),
8.55 (s, 1H); MS (FAB) m/z 536 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.30FN.sub.3O.sub.6.0.5H.sub.2O: C, 63.96; H, 5.74; N,
7.72; F, 3.49. Found: C, 64.11; H, 5.80; N, 7.39; F, 3.54.
Example 35
4-[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinyl]-
methoxy-3-methoxy benzoic acid
[0433] ##STR1660##
[0434] A mixture of
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetic acid (318 mg, 1
mmol), 2-(2-methoxy-4-ethoxycarbonyl)phenoxymethylpyrrolidine (279
mg, 1 mmol), EDC (288 mg, 1.5 mmol), and DMAP (122 mg, 1 mmol) in
DMF (20 mL) was stirred overnight. The mixture was poured into 1 N
HCl and the resulting solid was collected with suction. The solid
was dissolved in CHCl.sub.3 and dried over MgSO.sub.4. After
removal of the solvent, the residue was chromatographed on
silica-gel with CHCl.sub.3/MeOH (100:1, v/v) as eluent to give an
oil, which was dissolved in THF:MeOH (4:1, v/v, 10 mL). 0.25 N NaOH
(8 mL) was added to the solution and the resulting stirred mixture
was heated under reflux for 3 hr. The mixture was poured into 1 N
HCl. The resulting solid was collected with suction, dissolved in
CHCl.sub.3, dried over MgSO.sub.4, and evaporated. The residue was
recrystallized from CHCl.sub.3-n-hexane-ether to give 329 mg (60%)
39 as a white crystalline powder. mp 140-144; IR (KBr) 3338, 2956,
2875, 2607, 1709 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta.
1.95-2.25 (4H, m), 3.45-4.50 (12H, m), 6.66-8.15 (12H, m); MS (FAB)
m/z 552 (M.sup.++1).
Example 36
2-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methoxy]pyridine-5-carboxylic acid
[0435] ##STR1661##
[0436] To a stirred solution of 6-hydroxynicotinic acid (500 mg,
3.594 mmol) in benzene (8 mL) and MeOH (2 mL) was added dropwise
TMSCHN.sub.2 (1.97 mL, 3.953 mmol) at 0.degree. C., and the mixture
was stirred overnight at room temp. The reaction mixture was
quenched by the addition of AcOH, and the resulting mixture was
concentrated in vacuo. The residue was chromatographed on
silica-gel with n-hexane-EtOAc (1:3, v/v) as eluent to give 269.8
mg (49%) methyl 2-hydroxypyridine-5-carboxylate as a white
crystalline powder. IR (KBr) 3062, 1657, 1654, 1612, 1435, 1300,
1113, 775, 642 cm.sup.-1; .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
3.87 (s, 3H), 6.58 (d, J=9.8 Hz, 1H), 7.99 (dd, J=2.4, 9.8 Hz, 1H),
8.19 (d, J=2.4 Hz, 1H); MS (FAB) m/z 154 (M.sup.++1); Anal. Calcd
for C.sub.7H.sub.7NO.sub.3.1/4H.sub.2O: C, 53.33; H, 4.80; N, 8.89.
Found: C, 53.58; H, 4.65; N, 8.87.
[0437] To a stirred solution of methyl
2-hydroxypyridine-5-carboxylate (269.8 mg, 1.762 mmol),
N-tert-butoxycarbonylprolinol (354.6 mg, 1.762 mmol), and Ph.sub.3P
(554.6 mg, 2.114 mmol) in THF (10 mL) was slowly added DIAD (0.42
mL, 2.114 mmol) at room temp, and the resulting mixture was stirred
for 6 hr at 70.degree. C. The reaction mixture was concentrated and
the residue was chromatographed on silica-gel with n-hexane:EtOAc
(5:1, v/v) as eluent to give 262.5 mg (44%) methyl
2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy]pyridine-5-carboxylate
as an oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.47 (s, 9H),
1.85-1.98 (m, 4H), 3.37 (br s, 2H), 3.92 (s, 3H), 4.12-4.33 (br m,
2H), 4.4 (brs, 1H), 6.75 (m, 1H), 8.15 (m, 1H), 8.79 (m, 1H); MS
(FAB) m/z 337 (M.sup.++1).
[0438] To a stirred solution of methyl
2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy]pyridine-5-carboxylate
(262.5 mg, 0.870 mmol) in CH.sub.2Cl.sub.2 (5.3 mL) was added TFA
(1.1 mL) at 0.degree. C., and the resulting mixture was stirred at
room temp for 1 hr. The solvent was removed under a reduced
pressure and the residue was made basic by the addition of the 1N
NaOH, and extracted with CHCl.sub.3. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated under a
reduced pressure to afford 173.1 mg (94%) methyl
2-[(2-pyrrolidinyl)methoxy]pyridine-5-carboxylate as a pale
yellowish oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.49-1.58
(ddt, J=6.8, 8.8 Hz, 1H), 1.72-1.87 (m, 2H), 1.90-1.99 (m, 1H),
2.92-3.05 (m, 2H), 3.50-3.57 (ddd, J=4.4, 7.3, 15.1 Hz, 1H), 3.91
(s, 3H), 4.23 (dd, J=7.8, 10.7 Hz, 1H), 4.38 (dd, J=4.4, 10.7 Hz,
1H), 6.78 (d, J=8.8 Hz, 1H), 8.15 (dd, J=2.4, 8.8 Hz, 1H), 8.80 (d,
J=2.4 Hz, 1H); MS (FAB) m/z 237 (M.sup.++1).
[0439] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (351.7 mg,
0.732 mmol), methyl
2-[(2-pyrrolidinyl)methoxy]pyridine-5-carboxylate (173.0 mg, 0.732
mmol), Et.sub.3N (122.4 .mu.l, 0.878 mmol) in DMF (5.2 mL) was
stirred for 1 hr at room temp. The mixture was diluted with EtOAc,
washed with brine, and dried over Na.sub.2SO.sub.4. The solvent was
removed under a reduced pressure and the residue was
chromatographed on silica-gel with n-hexane:EtOAc (1:5, v/v) as
eluent to afford methyl
2-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methoxy]pyridine-5-carboxylate (338.4 mg, 87%) as an oil. To a
stirred solution of this compound in THF (5.6 mL) and H.sub.2O (1.4
mL) was added LiOH (45.7 mg, 1.91 mmol), and the reaction mixture
was stirred at room temp overnight. The mixture was diluted with
CHCl.sub.3, and treated with sat. NH.sub.4Cl, washed with brine,
dried over Na.sub.2SO.sub.4. The solvent was removed under a
reduced pressure, and the obtained crude solid was recrystallized
from n-hexane-Et.sub.2O--CHCl.sub.3-MeOH to afford 193.8 mg (59%)
40 as a white crystalline powder. mp 125-128; IR (KBr) 1716, 1600,
1533, 1255 cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta.
1.67-2.03 (m, 4H), 2.50 (s, 3H), 3.33-3.42 (m, 1H), 3.52 (m, 2H),
3.58 (d, J=4.4 Hz, 1H), 3.83 (s, 3H), 4.27-4.31 (m, 2H), 4.42-4.47
(m, 1H), 6.73 (d, J=7.8 Hz, 1H), 6.87-6.95 (m, 3H), 7.11-7.17 (m,
2H), 7.79 (d, J=8.3 Hz, 1H), 7.99 (d, J=8.3 Hz, 1H), 8.14 (dd,
J=2.0, 8.8 Hz, 1H), 8.46 (s, 1H), 8.56 (s, 1H), 8.69 (d, J=2.0 Hz,
1H), 13.06 (br s, 1H); MS (FAB) m/z 519 (M.sup.++1); Anal. Calcd
for C.sub.28H.sub.30N.sub.4O.sub.6.1/2H.sub.2O: C, 63.75; H, 5.92;
N, 10.62. Found: C, 63.61; H, 5.94; N, 10.27.
Example 37
3-methoxy-4-[2-[4-[N'-(2-methylphenyl)ureido]benzyl]-4-thiazolyl]methoxybe-
nzoic acid
[0440] ##STR1662##
[0441] To a stirred solution of phosphorous pentasulfide (27.4 g,
123.34 mmol) and freshly prepared anhydrous Na.sub.2S (4.8 g, 61.67
mmol) in THF (200 mL) was added p-nitrobenzyl cyanide (2.0 g, 12.33
mmol) at room temp. The resulting mixture was stirred for 17 hr at
room temp. The mixture was diluted with EtOAc and washed with 10%
K.sub.3PO.sub.4. The aqueous layer was extracted with
CH.sub.2Cl.sub.2. The extract was dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was chromatographed on
silica-gel with n-hexane:EtOAc (5:1 to 2:1, v/v) as eluent to give
1.53 g (64%) 4-nitrobenzyl carbothioamide as a pale yellow
crystalline material. IR (KBr) 1529, 1446, 1326, 1315, 858
cm.sup.-1; .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 4.15 (s, 2H),
7.51 (d, J=8.3 Hz, 2H), 8.24 (d, J=8.8 Hz, 2H); MS (FAB) m/z 197
(M.sup.++1); Anal. Calcd for C.sub.8H.sub.8N.sub.2O.sub.2S: C,
48.97; H, 4.11H; N, 14.28; S, 16.34. Found: C, 48.69; H, 4.06; N,
14.07; S, 16.10.
[0442] To a stirred solution of 4-nitrobenzylcarbothioamide (502.0
mg, 2.558 mmol) in EtOH (5 mL) was added 1,3-dichloro-2-propanone
(649.6 mg, 5.16 mmol) and the mixture was heated under reflux for 1
hr. The mixture was concentrated and the residue was diluted with
CHCl.sub.3. The solution was washed with 1N NaOH, brine, and dried
over Na.sub.2SO.sub.4. The solvent was removed under a reduced
pressure and the residue was chromatographed on silica-gel with
n-hexane:EtOAc (4:1, v/v) as eluent to afford 495.2 mg (72%)
4-[2-(4-nitrobenzyl)thiazolyl]methyl chloride as a pale yellow oil.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 4.43 (s, 2H), 4.68 (s,
2H), 7.23 (s, 1H), 7.49 (d, J=8.8 Hz, 2H), 8.20 (d, J=8.8 Hz, 2H);
MS (FAB) m/z 269 (M.sup.++1).
[0443] To a stirred solution of vanillic acid ethyl ester (308.0
mg, 1.570 mmol) and MeONa (89 mg, 1.570 mmol) in MeOH (6.5 ml) was
added a solution of 4-[2-(4-nitrobenzyl)thiazolyl]methyl chloride
(211.0 mg, 0.785 mmol) in MeOH (1.4 mL) at 0.degree. C. The
resulting mixture was stirred at room temp for 16 hr, and heated
under reflux for 1 day. The solvent was removed under a reduced
pressure and the residue was extracted with CHCl.sub.3. The extract
was washed with H.sub.2O, brine, and dried over Na.sub.2SO.sub.4.
The solvent was removed under a reduced pressure and the residue
was chromatographed on silica-gel with n-hexane:EtOAc (2:1, v/v) as
eluent to afford 201.7 mg (60%) ethyl
3-methoxy-4-[2-(4-nitrobenzyl)-4-thiazolyl]methoxybenzoate as a
pale yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.39 (t,
J=7.3 Hz, 3H), 3.93 (s, 3H), 4.36 (q, J=7.3 Hz, 2H), 4.44 (s, 2H),
5.31 (s, 2H), 6.97 (d, J=8.3 Hz, 1H), 7.28 (s, 1H), 7.48 (d, J=8.8
Hz, 2H), 7.57 (d, J=2.0 Hz, 1H), 7.64 (dd, J=2.0, 8.3 Hz, 1H), 8.20
(d, J=8.8 Hz, 2H); MS (FAB) m/z 429 (M.sup.++1).
[0444] A stirred solution of ethyl
3-methoxy-4-[2-(4-aminobenzyl)-4-thiazolyl methoxybenzoate (201.7
mg, 0.471 mmol) and 5% Pd/C (40 mg) in EtOH (8 mL) was hydrogenated
at 1 atm for 24 hr. The mixture was filtered and the filtrate was
concentrated. The residue was chromatographed on silica-gel with
n-hexane:EtOAc (1:1, v/v) as eluent to afford 87.8 mg (47%) ethyl
3-methoxy-4-[2-(4-aminobenzyl)-4-thiazolyl]methoxybenzoate as a
yellowish crystalline powder. .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. 1.38 (t, J=7.3 Hz, 3H), 3.93 (s, 3H), 4.21 (s, 2H), 4.35
(q, J=7.3 Hz, 2H), 5.30 (s, 2H), 6.66 (dd, J=2.0, 6.4 Hz, 2H), 6.97
(d, J=8.3 Hz, 1H), 7.09 (d, J=8.3 Hz, 2H), 7.18 (s, 1H), 7.56 (d,
J=2.0 Hz, 1H), 7.64 (dd, J=2.0, 8.3 Hz, 1H); MS (FAB) m/z 399
(M.sup.++1).
[0445] To a solution of ethyl
3-methoxy-4-[2-(4-aminobenzyl)-4-thiazolyl]methoxybenzoate (87.8
mg, 0.220 mmol) in THF (2.0 mL) was added triethylamine (30.5 ul,
0.220 mmol) and o-tolyl isocyanate (30 .mu.l), and the reaction
mixture was stirred at room temp for 21 hr. The reaction mixture
was poured into ice-water and the resulting precipitates filtered
off. The filtrate was extracted with CHCl.sub.3, washed with
H.sub.2O, and brine. The extract was dried over Na.sub.2SO.sub.4.
The solvent was removed under a reduced pressure to afford 110.4 mg
(94%) ethyl
3-methoxy-4-[2-[4-[N'-(2-methylphenyl)ureido]benzyl]-4-thiazolyl]methoxyb-
enzoate as a pale yellow crystalline powder. .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. 1.38 (t, J=7.3 Hz, 3H), 2.28 (s, 3H), 3.92 (s,
3H), 4.28 (s, 2H), 4.35 (q, J=7.3 Hz, 2H), 5.29 (s, 2H), 6.20 (s,
1H), 6.47 (s, 1H), 6.97 (d, J=8.8 Hz, 1H), 7.20 (s, 1H), 7.24 (s,
2H), 7.27 (s, 2H), 7.33 (d, J=8.3 Hz, 2H), 7.49 (d, J=7.3 Hz, 1H),
7.56 (d, J=2.0 Hz, 1H), 7.63 (dd, J=2.0, 8.3 Hz, 1H); MS (FAB) m/z
532 (M.sup.++1).
[0446] To a stirred solution of ethyl
3-methoxy-4-[2-[4-[N'-(2-methylphenyl)ureido]benzyl]-4-thiazolyl]methoxyb-
enzoate in THF (1.6 mL) and H.sub.2O (0.4 mL) was added LiOH (6.0
mg, 0.249 mmol), and the mixture was stirred at room temp for 1 hr,
and heated under reflux for 8 hr. The mixture was concentrated and
diluted with CHCl.sub.3. The solution was made basic by the
addition of 1N NaOH, The aqueous extract was acidified by the
addition of 1N HCl and extracted with CHCl.sub.3. The extract was
washed with brine and dried over Na.sub.2SO.sub.4. The solvent was
removed under a reduced pressure and the obtained crude solid was
recrystallized from n-hexane-EtOAc-EtOH to afford 59.6 mg (57%) 41
as a white crystalline powder. mp 243-245; IR (KBr) 3282, 1685,
1637, 1600, 1554, 1516, 1278, 1234, 763, 748 cm.sup.-1; .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 2.27 (s, 3H), 3.83 (s, 3H), 4.30
(s, 2H), 5.21 (s, 2H), 6.97 (t, J=7.3 Hz, 1H), 7.15-7.24 (m, 3H),
7.29 (d, J=8.8 Hz, 2H), 7.47 (d, J=8.3 Hz, 2H), 7.50 (s, 1H), 7.58
(d, J=8.3 Hz, 1H), 7.62 (s, 1H), 7.86 (d, J=7.8 Hz, 1H), 7.97 (s,
1H), 9.09 (s, 1H), 12.70 (br s, 1H); MS (FAB) m/z 504 (M.sup.++1);
Anal. Calcd for C.sub.27H.sub.25N.sub.3O.sub.5S.1/4H.sub.2O: C,
63.83; H, 5.06; N, 8.27. Found: C, 63.74; H, 4.99; N, 8.10.
Example 38
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-methyl-2-pyr-
rolidinyl]methoxy]-3-nitrobenzoic acid
[0447] ##STR1663##
[0448] To a stirred solution of the N-tert-butoxycarbonylproline
(6.00 g, 0.0279 mol) in MeOH:benzene (1:4, v/v) was added dropwise
2.0 M-n-hexane solution of TMSCHN.sub.2 (16.7 mL, 0.0334 mol) at
room temp. After the resulting solution was stirred for 1 hr at
room temp, the mixture was evaporated off in vacuo to afford 6.39 g
(100%) N-tert-butoxycarbonylproline methyl ester as a yellow syrup.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.41 (s, 9H), 1.85-1.98 (m, 4H),
2.21-2.28 (m, 2H), 3.72 (s, 3H), 4.29 (m, 1H).
[0449] To a stirred solution of diisopropylamine (2.02 mL, 0.0144
mol) in THF (30 mL) was added dropwise 1.59 M n-hexane solution of
n-BuLi (9.06 mL, 0.0144 mol) at minus 78.degree. C. for over 5 min.
The resulting solution was stirred for 20 min at minus 78.degree.
C. To the solution was added dropwise N-tert-butoxycarbonylproline
methyl ester (3.00 g, 0.0131 mmol) in THF (30 mL) at minus
78.degree. C. for over 5 min. The resulting solution was stirred
for 10 min at minus 78.degree. C. To the solution was added
dropwise MeI (0.900 mL, 0.0144 mol) at minus 78.degree. C. The
resulting solution was stirred for 30 min at minus 78.degree. C.
The solution was quenched by the addition of sat. NH.sub.4Cl. The
resulting mixture was extracted with CHCl.sub.3. The extract was
washed with water, dried over Na.sub.2SO.sub.4, and evaporated in
vacuo to afford 3.20 g (q.y.) N-tert-butoxycarbonyl-2-methylproline
methyl ester as a yellow syrup. .sup.1H-NMR (CDCl.sub.3) .delta.
1.33 (s, 9H), 1.38 (s, 3H), 1.72-2.20 (m, 4H) 3.27-3.59 (m, 2H)
3.63 (d, J=6.3 Hz, 3H).
[0450] To a stirred solution of
N-tert-butoxycarbonyl-2-methylproline methyl ester (3.20 g, 0.0131
mol) in THF (20 mL) was added 1N NaOH (15.7 mL) at room temp. After
the resulting mixture was stirred for 24 hr, the mixture was
diluted with water and washed with EtOAc. The separated aqueous
layer was acidified by the addition of 1N HCl, and extracted with
EtOAc. The extract was dried over Na.sub.2SO.sub.4 and evaporated
in vacuo to afford 1.71 g (57%)
N-tert-butoxycarbonyl-2-methylproline as a yellow syrup.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.42 (s, 9H), 1.48 (s, 3H),
1.88-2.31 (m, 4H), 3.34-3.57 (m, 2H), 9.35 (br s, 1H)
[0451] To a stirred solution of
N-tert-butoxycarbonyl-2-methylproline (1.10 g, 4.80 mmol) in THF
(20 mL) was added dropwise BH.sub.3--SMe.sub.2 (0.546 mL, 5.76
mmol) at room temp. After the resulting mixture was stirred for 6
hr at 80.degree. C., the mixture was evaporated in vacuo. The
residue was diluted with MeOH, washed with n-hexane (3.times.), and
evaporated in vacuo to afford 0.648 g (60%)
N-tert-butoxycarbonyl-2-hydroxymethyl-2-methylpyrrolidine as a
yellow syrup. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47 (s, 9H),
1.76-2.05 (m, 4H), 3.28-3.48 (m, 2H), 3.66 (m, 2H, d).
[0452] To a stirred solution of
N-tert-butoxycarbonyl-2-hydroxymethyl-2-methylpyrrolidine (0.648 g,
3.01 mmol), methyl 4-hydroxy-3-nitrobenzoate (0.593 g, 3.01 mmol),
and Ph.sub.3P (0.868 g, 3.31 mmol) in THF (10 mL) was added
dropwise DIAD (95%) (0.686 mL, 3.31 mmol) at 0.degree. C. After the
resulting mixture was stirred for 24 hr at 80.degree. C., the
mixture was evaporated in vacuo. The residue was diluted with
CH.sub.2Cl.sub.2 (5 mL) and added TFA (5 mL). After the resulting
mixture was stirred for 2 hr at room temp, the mixture was
evaporated in vacuo. The residue was diluted with 0.5N HCl and
extracted with CHCl.sub.3. The aqueous layer was neutralized with
sat. NaHCO.sub.3, and extracted with CHCl.sub.3. The extract was
dried over Na.sub.2SO.sub.4 and evaporated in vacuo to afford 0.188
g (21%) methyl 3-nitro-4-(2-methyl-2-pyrrolidinylmethoxy)benzoate
as a yellow syrup.
[0453] A mixture of methyl
3-nitro-4-(2-methyl-2-pyrrolidinylmethoxy)benzoate (0.188 g, 0.920
mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(0.289 g, 0.920 mmol), HOBt (0.149 g, 1.10 mmol), DMAP (11.2 mg,
0.0920 mmol) and EDC (0.211 g, 1.10 mmol) in DMF (5 mL) was stirred
for 14 hr at room temp. EtOAC was added to the mixture and the
solution was washed successively with 0.5 N HCl, sat. NaHCO.sub.3,
and brine. The organic layer was dried over Na.sub.2SO.sub.4 and
evaporated in vacuo to afford 0.489 g (q.y.) methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-meth-
yl-2-pyrrolidinyl]methoxy]-3-nitrobenzoate as a yellow crystalline
material. .sup.1H-NMR (CDCl.sub.3) .delta. 1.26 (d, J=5.9 Hz),
1.85-4.50 (m, 10H), 2.30 (s, 3H), 3.67 (s, 2H), 3.92 (s, 3H), 6.36
(s, 2H), 6.75-7.52 (m, 7H), 8.02 (d, J=7.8 Hz, 1H), 8.15 (d, J=8.8
Hz, 1H), 8.47 (s, 1H).
[0454] A stirred mixture of
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-methyl-2-pyr-
rolidinyl]methoxy]-3-nitrobenzoate (0.489 g, 0.828 mmol) in MeOH (5
mL) and 1N NaOH (1.24 mL) was heated under reflux for 2 hr After
cooling, the mixture was diluted with water and extracted with
CHCl.sub.3. The aqueous layer was acidified with 1N HCl, and
extracted with CHCl.sub.3. The extract was dried over
Na.sub.2SO.sub.4 and evaporated in vacuo to afford 0.366 g (99%) 42
as a yellow crystalline material.
Example 39
4-[4-hydroxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyr-
rolidinylmethoxy]-3-methoxybenzoic acid
[0455] ##STR1664##
[0456] A stirred mixture of
4-[4-benzyloxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2--
pyrrolidinylmethoxy]-3-methoxybenzoate (440 mg, 0.645 mmol) and 5%
Pd/C (400 mg) in AcOH:EtOH (1:1, v/v, 100 mL) was hydrogenated at 1
atm for 5 hr. The mixture was filtered to remove the catalyst and
the filtrate was concentrated in vacuo. The residue was
chromatographed on silica-gel with CHCl.sub.3:EtOH (10:1, v/v) as
eluent to give 90 mg (24%) ethyl
4-[4-hydroxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-py-
rrolidinylmethoxy]-3-methoxybenzoate as a pale yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.39 (3H, t, J=7.3 Hz), 2.04-2.37
(total 5H, m), 3.44-4.70 (16H, series of m), 6.63 (1H, s),
6.70-6.80 (2H, m), 6.84 (1H, d, J=8.3 Hz), 7.11 (1H, t, J=7.8 Hz),
7.20-7.24 (3H, m), 7.45 (1H, d, J=2.0 Hz), 7.59 (2H, dd, J=8.3, 2.0
Hz), 8.01 (1H, d, J=7.8 Hz).
[0457] A stirred mixture of ethyl
4-[4-hydroxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-py-
rrolidinylmethoxy]-3-methoxybenzoate (90 mg, 0.152 mmol) in 0.25 N
NaOH (5 mL, 1.25 mmol) and THF (5 mL) was heated under reflux
overnight. The mixture was poured into ice-1 N HCl (200 mL). The
precipitate was collected with suction and recrystallized from
CHCl.sub.3-MeOH-n-hexane to give 40 mg (47%) 43 as a colorless
amorphous solid. .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.92-2.11 (2H,
m), 2.24 (3H, s), 3.31-5.07 (14H, series of m), 6.73 (1H, d, J=8.3
Hz), 6.84 (1H, s), 6.93 (1H, t, J=7.8 Hz), 7.01-7.17 (3H, m), 7.44
(1H, s), 7.52 (1H, d, J=8.8 Hz), 7.79 (1H, d, J=8.3 Hz), 7.99 (1H,
d, J=7.8 Hz), 8.46 (1H, s), 8.55 (1H, s), 12.67 (1H, br s); MS
(FAB) m/z 564 (M.sup.++1).
Example 40
(2S,4R)-3-amino-4-[4-hydroxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phe-
nylacetyl]-2-pyrrolidinyl]methoxybenzoic acid
[0458] ##STR1665##
[0459] To a stirred solution of
(2S,4R)-4-benzyloxy-1-(tert-butoxycarbonyl)-2-prolinol (891 mg, 2.9
mmol), methyl 4-hydroxy-3-nitrobenzoate (572 mg, 2.9 mmol), and
PPh.sub.3 (839 mg, 3.2 mmol) in THF (6 mL) was added DIAD (630 mL,
3.2 mmol) and the mixture was heated under reflux overnight. After
removal of the solvent, the residue was chromatographed on
silica-gel with n-hexane:EtOAc (1:1) and toluene:EtOAc (10:1, v/v)
as eluent to give 700 mg (50%) methyl
(2S,4R)-4-[4-benzyloxy-1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-n-
itrobenzoate as a pale yellow oil.
[0460] To a stirred solution of methyl
(2S,4R)-4-[4-benzyloxy-1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-n-
itrobenzoate (681 mg, 1.4 mmol) in CH.sub.2Cl.sub.2 (2 mL) was
added TFA (2 mL), and the resulting mixture was stirred for 2 hr.
After the reaction mixture was concentrated, the residue was made
basic by the addition of sat. NaHCO.sub.3 and extracted with
CHCl.sub.3. The extract was washed with H.sub.2O, dried over
MgSO.sub.4, and evaporated to give 511 mg (95%) methyl
(2S,4R)-4-[4-benzyloxy-2-pyrrolidinyl]methoxy-3-nitrobenzoate as a
yellow oil.
[0461] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (409 mg,
1.3 mmol), methyl
(2S,4R)-4-(benzyloxy-(2-pyrrolidinyl)methoxy-3-nitrobenzoate (502
mg, 1.3 mmol), EDC (383 mg, 2 mmol), and DMAP (159 mg, 1.3 mmol) in
DMF (20 mL) was stirred for 3 days. The mixture was poured into 1 N
HCl and the resulting precipitate was collected with suction. The
residue was dissolved in CHCl.sub.3 and dried over MgSO.sub.4.
After removal of solvent, the residue was chromatographed on
silica-gel with CHCl.sub.3:MeOH (200:1, v/v) as eluent to give 680
mg (91%) methyl
(2S,4R)-4-[4-benzyloxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylac-
etyl]-2-pyrrolidinyl]methoxy-3-nitrobenzoate as a white amorphous
solid.
[0462] A solution of methyl
(2S,4R)-4-[4-benzyloxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylac-
etyl]-2-pyrrolidinyl]methoxy-3-nitrobenzoate (676 mg, 0.99 mmol)
and 5% Pd--C (1 g) in EtOH:AcOH (1:1, v/v, 30 mL) was hydrogenated
at 1 atm for 6 hr. The mixture was filtered and the filtrate was
evaporated to give an oil, which was made basic by the addition of
sat. NaHCO.sub.3. The mixture was extracted with EtOAc. The extract
was washed with brine, dried over MgSO.sub.4, and evaporated. The
residue was recrystallized from CHCl.sub.3-EtOH-n-hexane as eluent
to give 120 mg (22%) 44 as a pale yellow crystalline powder. MS
(FAB) m/z 549 (M.sup.++1)
Example 41
4-[[4-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyr-
rolidinyl]methoxy]-3-methoxybenzoic acid
[0463] ##STR1666##
[0464] A stirred mixture of ethyl
4-[4-benzyloxy-1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy-3-methoxybe-
nzoate (1.189 g, 2.449 mmol) and 5% Pd--C (240 mg) in EtOH (10 mL)
was hydrogenated overnight at room temp. The mixture was filtered
to remove the catalyst and the filtrate was concentrated in vacuo
to give ethyl
4-[1-(tert-butoxycarbonyl)-4-hydroxy-2-pyrrolidinyl]methoxy-3-methoxybenz-
oate (735.3 mg, 76%) as a pale yellow oil. To a stirred cold (minus
78.degree. C.) solution of DAST (0.491 mL, 3.718 mmol) in
CH.sub.2Cl.sub.2 (7.4 mL) was added dropwise a solution of this
compound in CH.sub.2Cl.sub.2 (2 mL), and the resulting mixture was
stirred overnight. The mixture was quenched with water and
extracted with CHCl.sub.3. The extract was washed with brine and
dried over Na.sub.2SO.sub.4. The solvent was removed under a
reduced pressure and the residue was chromatographed on silica-gel
with n-hexane:EtOAc (3:1, v/v) as eluent to afford 418.7 mg (57%)
ethyl
4-[1-(tert-butoxycarbonyl)-4-fluoro-2-pyrrolidinyl]methoxy-3-methoxybenzo-
ate as an oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.39 (t,
J=7.3 Hz, 3H), 1.49 (s, 9H), 2.16 (br m, 1H), 2.58 (dd, J=15.6,
19.0 Hz, 1H), 3.60-3.75 (m, 2H), 3.91 (s, 3H), 3.97 (t, J=9.3 Hz,
1H), 4.35 (q, J=7.3 Hz, 2H), 4.33-4.53 (m, 2H), 5.25 (d, J=52.7 Hz,
1H), 7.04 (dd, J=7.8, 56.2 Hz, 1H), 7.55 (s, 1H), 7.65 (br s, 1H);
MS (FAB) m/z 398 (M.sup.++1).
[0465] To a stirred solution of ethyl
4-[1-(tert-butoxycarbonyl)-4-fluoro-2-pyrrolidinyl]methoxy-3-methoxybenzo-
ate (482.2 mg, 1.213 mmol) in CH.sub.2Cl.sub.2 (10.0 mL) was added
TFA (1.9 mL) at 0.degree. C., and the mixture was stirred at room
temp for 2 hr. The solvent was removed under a reduced pressure and
the residue was made basic by the addition of 1N NaOH and extracted
with CHCl.sub.3. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated under a reduced pressure to
afford 348.7 mg (97%) ethyl
4-(4-fluoro-(2-pyrrolidinyl)methoxy-3-methoxybenzoate as a brownish
oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.39 (t, J=6.8 Hz,
3H), 1.97 (ddt, J=1.5, 5.4, 14.7 Hz, 1H), 2.27 (dddd, J=5.9, 8.8,
14.7, 32.7 Hz, 1H), 3.02 (ddd, J=3.9, 13.1, 35.2 Hz, 1H), 3.36 (dd,
J=12.7, 21.5 Hz, 1H), 3.65 (m, 1H), 3.90 (s, 3H), 4.09 (m, 1H),
4.35 (q, J=6.8 Hz, 2H), 5.17, 5.31 (br m each, 1H), 6.90 (d, J=8.3
Hz, 1H), 7.75 (d, J=2.0 Hz, 1H), 7.65 (dd, J=2.0, 8.3 Hz, 1H); MS
(FAB) m/z 298 (M.sup.++1).
[0466] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (404.0 mg,
0.840 mmol), ethyl
4-(4-fluoro-(2-pyrrolidinyl)methoxy-3-methoxybenzoate (250.0 mg,
0.840 mmol), Et.sub.3N (141 .mu.l, 1.009 mmol) in DMF (4.0 mL) was
stirred for 1 hr at room temp. The mixture was diluted with EtOAc,
washed with water, brine, and dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure and the residue was
chromatographed on silica-gel with n-hexane:EtOAc (1:3, v/v) to
afford 502 mg (q.y.) of ethyl
4-[[4-fluor-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-
-2-pyrrolidinyl]methoxy]-3-methoxybenzoate as a yellow oil. To a
stirred solution of this compound in THF (8.0 mL) and H.sub.2O (2.0
mL) was added LiOH (60.4 mg, 2.520 mmol), and the mixture was
stirred at room temp. overnight, and 50.degree. C. for 1 day. The
mixture was diluted with CHCl.sub.3, and extracted with 1N NaOH,
The aqueous layer was acidified by the addition of 1N HCl and
extracted with CHCl.sub.3. The extract was washed with brine and
dried over Na.sub.2SO.sub.4. The solvent was removed under a
reduced pressure and the obtained crude solid was recrystallized
from EtOAc--CHCl.sub.3-EtOH-n-hexane to afford 294.8 mg (62%) 45 as
a white crystalline powder. IR (KBr) 2958, 2937, 1687, 1601, 1531,
1454, 1419, 1267, 1214, 1029 cm.sup.-1; .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.86-2.09 (m, 5H), 2.06 (s, 3H), 2.25 (s,
3H), 3.47-3.67 (m, 6H), 3.76 (s, 3H), 4.05-4.12 (m, 2H), 4.30-4.31
(m, 1H), 6.51 (s, 1H), 6.55 (s, 1H), 6.73-6.95 (m, 2H), 7.11-7.17
(m, 2H), 7.64 (s, 1H), 7.79 (d, J=7.8 Hz, 1H), 7.99 (d, J=7.8 Hz,
1H), 8.47 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 566 (M.sup.++1);
Anal. Calcd for C.sub.30H.sub.32FN.sub.3O.sub.7.1/2H.sub.2O: C,
62.71; H, 5.79; F, 3.31; N, 7.31. Found: C, 63.13; H, 6.17; F,
3.12; N, 7.04.
Example 42
3-acetylamino-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-
-pyrrolidinyl methoxy]benzoic acid
[0467] ##STR1667##
[0468] A solution of
3-amino-4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrr-
olidinylmethoxy]benzoic acid (130 mg, 0.244 mmol) and DMAP (2.9 mg,
0.0244 mmol) in pyridine (5 mL) and acetic anhydride (5 mL) was
stirred for 2 hr at room temp. The mixture was evaporated off in
vacuo (excess acetic anhydride was azeotropically removed with
toluene). Water was added to the residue, and extracted with
CHCl.sub.3. The extract was dried over Na.sub.2SO.sub.4 and
evaporated in vacuo. The residue was chromatographed on silica-gel
with MeOH:CHCl.sub.3 (1:15 to 1:1, v/v) as eluent to afford 29 mg
(21%) 46 as a white crystalline material. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.80-2.30 (m, 4H), 2.04 (s, 3H), 2.26 (s,
3H), 3.33 (s, 3H), 3.40-4.80 (m, 7H), 6.59 (s, 1H), 6.74 (d, J=8.3
Hz, 1H), 6.79 (d, J=8.8 Hz, 1H), 7.07-7.57 (m, 6H), 7.75 (d, J=8.8
Hz, 1H), 8.07 (d, J=8.3 Hz, 1H), 8.41 and 8.96 (each s, each 1H);
MS (FAB) m/z 575 (M.sup.++1).
Example 43
3-chloro-2-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyr-
rolidinyl]methoxy]pyridine-5-carboxylic acid
[0469] ##STR1668##
[0470] To a stirred solution of
3-chloro-2-hydroxypyridine-5-carboxylic acid (1 g, 5.762 mmol) in
benzene (16 mL) and MeOH (4 mL) was added dropwise TMSCHN.sub.2
(3.17 mL, 6.338 mmol) at 0.degree. C., and the resulting mixture
was stirred overnight at room temp. The reaction mixture was
quenched by the addition of AcOH and the mixture was evaporated
off. The residue was suspended in water and precipitate was
collected. The crude solid was washed with Et.sub.2O, and dried
under a reduced pressure to give 728.1 mg (67%) methyl
3-chloro-2-hydroxypyridine-5-carboxylate as a white crystalline
powder. IR (KBr) 1655, 1282, 1245, 769 cm.sup.-1; .sup.1H-NMR (400
MHz, DMSO-d.sub.6) .delta. 3.79 (s, 3H), 8.01 (s, 1H), 8.06 (s,
1H); MS (FAB) m/z 188 (M.sup.++1); Anal. Calcd for
C.sub.7H.sub.6ClNO.sub.3: C, 44.82; H, 3.22; Cl, 18.90; N, 7.47.
Found: C, 44.74; H, 3.22; Cl, 19.00; N, 7.34.
[0471] To a stirred solution of methyl
3-chloro-2-hydroxypyridine-5-carboxylate (300 mg, 1.599 mmol),
N-tert-butoxycalbonylprolinol (321.9 mg, 1.599 mmol), and Ph.sub.3P
(503 mg, 1.919 mmol) in THF (3 mL) was slowly added DIAD (378
.mu.l, 1.919 mmol) at room temp, and the mixture was stirred for 13
hr at 70.degree. C. The mixture was concentrated and the residue
was chromatographed on silica-gel with n-hexane-EtOAc (3:1, v/v) as
eluent to give 235.6 mg (40%) methyl
3-chloro-2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy]pyridine-5-ca-
rboxylate as a pale yellowish oil. .sup.1H-NMR (400 MHz,
CDCl.sub.3) .delta. 1.46 (s, 9H), 1.87 (m, 1H), 2.05 (br s, 3H),
3.43 (br s, 2H), 3.92 (s, 3H), 4.17, 4.26 (br s each, 1H),
4.45-4.51 (m, 1H), 4.50 (s, 1H), 8.21 (s, 1H), 8.67 (d, J=2.0 Hz,
1H); MS (FAB) m/z 371 (M.sup.++1).
[0472] To a stirred solution of methyl
3-chloro-2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxy]pyridine-5-ca-
rboxylate (235.6 mg, 0.635 mmol) in CH.sub.2Cl.sub.2 (5.0 mL) was
added TFA (1.0 mL) at 0.degree. C., and the reaction mixture was
stirred at room temp for 2 hr. The solvent was removed under a
reduced pressure. The residue was made basic by the addition of 1N
NaOH and extracted with CHCl.sub.3. The extract was dried over
Na.sub.2SO.sub.4, concentrated under a reduced pressure to afford
172.3 mg (q.y.) methyl
3-chloro-2-[(2-pyrrolidinyl)methoxy]pyridine-5-carboxylate as a
pale yellowish oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.55-1.63 (m, 1H), 1.76-1.99 (m, 3H), 2.93-2.99 (m, 1H), 3.02-3.08
(m, 1H), 3.57-3.62 (m, 1H), 3.92 (s, 3H), 4.33 (dd, J=7.3, 10.7 Hz,
1H), 4.44 (dd, J=4.4, 10.7 Hz, 1H), 8.21 (d, J=2.0 Hz, 1H), 8.67
(d, J=2.0 Hz, 1H); MS (FAB) m/z 271 (M.sup.++1).
[0473] The mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (317.0 mg,
0.660 mmol), methyl
3-chloro-2-[(2-pyrrolidinyl)methoxy]pyridine-5-carboxylate (172.0
mg, 0.635 mmol), Et.sub.3N (105 ul, 0.756 mmol) in DMF (2.0 mL) was
stirred for 1 hr at room temp. The mixture was diluted with EtOAc,
washed with brine, and dried over Na.sub.2SO.sub.4. The solvent was
removed under a reduced pressure to afford methyl
3-chloro-2-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido)phenylacetyl]-2-py-
rrolidinyl]methoxy]pyridine-5-carboxylate as a brownish oil. To a
stirred solution of this compound in THF (6.0 mL) and H.sub.2O (2.0
mL) was added LiOH (45.3 mg, 1.89 mmol), and the reaction mixture
was stirred for 5 hr at room temp. The mixture was diluted with
n-hexane and extracted with 1N-NaOH, The aqueous layer was
acidified by the addition of 1N HCl and extracted with CHCl.sub.3.
The extract was washed with brine, dried over Na.sub.2SO.sub.4. The
solvent was removed under a reduced pressure and the obtained crude
solid was recrystallized from n-hexane-EtOAc-EtOH to afford 242.2
mg (70%) 47 as an orange crystalline powder. mp 122-125; IR (KBr)
3354, 1709, 1593, 1535, 1454, 1257 cm.sup.-1; .sup.1H-NMR (400 MHz,
DMSO-d.sub.6) .delta. 1.67-2.03 (m, 4H), 2.50 (s, 3H), 3.33-3.42
(m, 1H), 3.52 (m, 2H), 3.58 (d, J=4.4 Hz, 1H), 3.83 (s, 3H),
4.27-4.31 (m, 2H), 4.42-4.47 (m, 1H), 6.73 (d, J=7.8 Hz, 1H),
6.87-6.95 (m, 3H), 7.11-7.17 (m, 2H), 7.79 (d, J=8.3 Hz, 1H), 7.99
(d, J=8.3 Hz, 1H), 8.14 (dd, J=2.0, 8.8 Hz, 1H), 8.46 (s, 1H), 8.56
(s, 1H), 8.69 (d, J=2.0 Hz, 1H), 13.06 (br s, 1H); MS (FAB) m/z 553
(M.sup.++1); Anal. Calcd for C.sub.28H.sub.29ClN.sub.4O.sub.6: C,
60.81; H, 5.29; N, 10.31. Found: C, 60.98; H, 5.50; N, 9.46.
Example 44
2-[[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinyl-
]methoxy]pyridine-5-carboxylic acid
[0474] ##STR1669##
[0475] To a stirred solution of 6-hydroxynicotinic acid (2 g, 14.38
mmol) in benzene (32 mL) and MeOH (8 mL) was added dropwise
TMSCHN.sub.2 (1.97 mL, 3.953 mmol) at 0.degree. C., and the
resulting mixture was stirred for 2 hr at room temp. The mixture
was quenched by the addition of AcOH and concentrated in vacuo. The
residue was suspended in water and the solid was collected. The
crude solid was washed with Et.sub.2O, and dried in vacuo to give
1.566 g (71%) methyl 2-hydroxypyridine-5-carboxylate as a pale
brown crystalline powder. IR (KBr) 1655, 1645, 1610, 1433, 1300,
1113, 777, 642 cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6)
.delta. 3.77 (s, 3H), 6.37 (d, J=9.8 Hz, 1H), 7.99 (dd, J=2.4, 9.8
Hz, 1H), 8.03 (d, J=2.4 Hz, 1H); MS (FAB) m/z 154 (M.sup.++1);
Anal. Calcd for C.sub.7H.sub.7NO.sub.3: C, 54.90; H, 4.61; N, 9.15.
Found: C, 54.89; H, 4.60; N, 9.13.
[0476] To a stirred solution of methyl
2-hydroxypyridine-5-carboxylate (1.00 g, 6.529 mmol),
N-tert-butoxycarbonylprolinol (1.31 g, 6.529 mmol), and Ph.sub.3P
(2.06 g, 7.836 mmol) in THF (10 mL) was added DIAD (1.54 mL, 7.836
mmol) at room temp, and the resulting mixture was stirred for 13 hr
at 70.degree. C. The mixture was concentrated and the residue was
chromatographed on silica-gel with n-hexane:EtOAc (3:1, v/v) as
eluent to give 712.3 mg (32%) methyl
2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methoxylpyridine-5-carboxylate
as a pale yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.47 (s, 9H), 1.85-1.98 (m, 4H), 3.37 (br s, 2H), 3.92 (s, 3H),
4.12-4.33 (br m, 2H), 4.48 (br s, 2H), 6.75 (m, 1H), 8.15 (m, 1H),
8.79 (m, 1H); MS (FAB) m/z 337 (M.sup.++1).
[0477] To a stirred solution of methyl
2-[[1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methyloxy]pyridine-5-carboxyla-
te (232.3 mg, 0.691 mmol) in CH.sub.2Cl.sub.2 (4.6 mL) was added
TFA (0.9 mL) at 0.degree. C., and the reaction mixture was stirred
at room temp for 2 hr. The solvent was removed under a reduced
pressure and the residue was made basic by the addition of 1N NaOH,
The aqueous solution was extracted with CHCl.sub.3, washed with
brine, and the dried over Na.sub.2SO.sub.4. The solvent was
evaporated under a reduced pressure to afford 146.2 mg (90%) methyl
2-(2-pyrrolidinyl)methoxypyridine-5-carboxylate as an oil.
.sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.49-1.58 (m, 1H),
1.72-2.18 (m, 3H), 2.92-3.05 (m, 2H), 3.50-3.57 (m, 1H), 3.91 (s,
3H), 4.23 (dd, J=8.0, 10.7 Hz, 1H), 4.38 (dd, J=4.4, 10.3 Hz, 1H),
6.78 (d, J=8.8 Hz, 1H), 8.15 (dd, J=2.4, 8.8 Hz, 1H), 8.80 (d,
J=2.4 Hz, 1H); MS (FAB) m/z 237 (M.sup.++1).
[0478] The mixture of pentafluorophenyl
4-[N'-(2-fluorophenyl)ureido]-3-methoxy-phenylacetate (314.8 mg,
0.650 mmol), methyl
2-[(2-pyrrolidinyl)methoxy]pyridine-5-carboxylate (146.2 mg, 0.619
mmol), Et.sub.3N (103 ul, 0.743 mmol) in DMF (1.5 mL) was stirred
for 1 hr at room temp. The mixture was diluted with Et.sub.2O,
washed with brine, and dried over Na.sub.2SO.sub.4. The solvent was
removed under a reduced pressure to afford methyl
2-[[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidiny-
l]methoxy]pyridine-5-carboxylate as a crude pale yellow oil.
[0479] To a stirred solution of this compound in THF (6.0 mL) and
H.sub.2O (2.0 mL) was added LiOH (44.5 mg, 1.857 mmol), and the
reaction mixture was stirred for 17 hr at room temp. The mixture
was diluted with n-hexane and made basic by the addition of 1N
NaOH, The aqueous layer was acidified by 1N HCl and extracted with
CHCl.sub.3. The extract was washed with brine, and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure
and the obtained crude solid was recrystallized from
n-hexane-EtOAc-EtOH to afford 202.5 mg (63%) 48 as a white
crystalline powder. IR (KBr) 1602, 1537, 1456, 1265, 752 cm.sup.-1;
H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.67-2.03 (m, 4H), 2.50 (s,
3H), 3.33-3.42 (m, 1H), 3.52 (m, 2H), 3.58 (d, J=4.4 Hz, 1H), 3.83
(s, 3H), 4.27-4.31 (m, 2H), 4.42-4.47 (m, 1H), 6.73 (d, J=7.8 Hz,
1H), 6.87-6.95 (m, 3H), 7.11-7.17 (m, 2H), 7.79 (d, J=8.3 Hz, 1H),
7.99 (d, J=8.3 Hz, 1H), 8.14 (dd, J=2.0, 8.8 Hz, 1H), 8.46 (s, 1H),
8.56 (s, 1H), 8.69 (d, J=2.0 Hz, 1H), 13.06 (br s, 1H); MS (FAB)
m/z 523 (M.sup.++1); Anal. Calcd for
C.sub.27H.sub.27FN.sub.4O.sub.6.1/2H.sub.2O: C, 61.01; H, 5.31; N,
10.54. Found: C, 61.52; H, 5.39; N, 10.01.
Example 45
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinylm-
ethyl]-1-piperazinyl acetic acid
[0480] ##STR1670##
[0481] To a stirred suspension of 1-benzylpiperazine (5 g, 28.4
mmol) and K.sub.2CO.sub.3 (5.89 g, 42.6 mmol) in DMF (30 mL) was
added ethyl bromoacetate (4.74 g, 28.4 mmol) at room temp. The
resulting mixture was stirred for a further 3 hr. The mixture was
diluted with EtOAc (300 mL), washed with brine (2.times.100 mL),
dried over MgSO.sub.4 and evaporated. The residue was
chromatographed on silica-gel with CHCl.sub.3:EtOH (10:1, v/v) as
eluent to give 7.45 g (q.y.) ethyl 4-benzyl-1-piperazinylacetate as
a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.27 (3H, t, J=7.3
Hz), 2.88-2.96 (8H, m), 3.20 (2H, s), 3.52 (2H, s), 4.18 (2H, q,
J=7.3 Hz), 7.22-7.32 (5H, m).
[0482] A stirred solution of ethyl 4-benzyl-1-piperazinylacetate
(2.00 g, 7.62 mmol) and 5% Pd/C (2 g) in AcOH:EtOH (1:1, 40 mL) was
hydrogenated at 1 atm for 8 hr. The mixture was filtered and the
filtrate was concentrated in vacuo. The residue was made basic by
the addition of saturated NaHCO.sub.3 and extracted with CHCl.sub.3
(2.times.200 mL). The combined extracts were dried over
K.sub.2CO.sub.3 and evaporated to give 1.16 g (88%) ethyl
1-piperazinylacetate as a yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.26-1.30 (3H, m), 1.67 (1H, br s), 2.55 (4H, m), 2.92-2.96
(4H, m), 3.19-3.20 (2H, m), 4.16-4.22 (2H, m).
[0483] To a stirred solution of N-Boc-L-prolinol (1.00 g, 5.02
mmol) and ethyl 1-piperazinyl acetate (864 mg, 5.02 mmol) in
MeOH:AcOH (10:1, v/v, 11 mL) was added NaBH.sub.3CN (664 mg, 10.0
mmol) at room temp. After being stirred overnight, the mixture was
poured into ice water (100 mL) and made basic by the addition of
NaHCO.sub.3. The mixture was extracted with CHCl.sub.3 (2.times.200
mL). The combined extracts were dried over Na.sub.2CO.sub.3 and
evaporated. The residue was chromatographed on silica-gel with
CHCl.sub.3:EtOH (10:1, v/v) as eluent to give 1.20 g (67%) ethyl
4-[1-(tert-butoxy
carbonyl)-2-pyrrolidinylmethyl]-1-piperazinylacetate as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.27 (3H, t, J=7.3 Hz),
1.46-1.47 (9H, m), 1.79-3.96 (total 19H series of m), 4.19 (2H, q,
J=7.3 Hz).
[0484] A mixture of ethyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethyl]-1-piperazinylacetate
(1.20 g, 3.38 mmol) in TFA (5 mL) and CH.sub.2Cl.sub.2 (5 mL) was
stirred overnight. After removal of the solvent, the residue was
made basic by the addition of sat. NaHCO.sub.3. The mixture was
extracted with CHCl.sub.3 (2.times.200 mL). The combined extracts
were dried over Na.sub.2CO.sub.3 and evaporated to give 386 mg
(45%) ethyl 4-(2-pyrrolidinylmethyl)-1-piperazinylacetate as a
yellow oil. MS (FAB) 256 (M.sup.++1).
[0485] To a stirred solution of ethyl
4-(2-pyrrolidinylmethyl)-1-piperazinylacetate (380 mg, 1.49 mmol)
and 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (468
mg, 1.49 mmol) in DMF (10 mL) was added EDC.HCl (428 mg, 2.24
mmol), HOBt, and DMAP (cat.). After being stirred overnight, the
mixture was diluted with EtOAc (300 mL), washed with brine
(2.times.200 mL), and dried over MgSO.sub.4. After removal of the
solvent, the residue was chromatographed on silica-gel with
CHCl.sub.3:EtOH (9:1, v/v) as eluent to give 257 mg (31%) ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
methyl]-1-piperazinylacetate as a yellow foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.24-1.29 (3H, m), 1.69-4.24 (total 29H,
series of m), 6.41 (1H, m), 6.81 (2H, m), 7.13-7.26 (4H, m), 7.52
(1H, d, J=7.3 Hz), 8.04 (1H, d, J=8.3 Hz).
[0486] To a stirred solution of ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
methyl]-1-piperazinylacetate (250 mg, 0.453 mmol) in THF (4 mL) was
added 0.25 N NaOH (3.6 mL). The resulting mixture was stirred
overnight. The mixture was adjusted to pH 7.5 by the addition of 1
N HCl and extracted with CHCl.sub.3:MeOH (4:1, 3.times.100 mL). The
combined extracts were dried over MgSO.sub.4 and evaporated. The
crude solid was recrystallized from CHCl.sub.3-MeOH-n-hexane to
give 40 mg (17%) 49 as a colorless crystalline powder. mp
160-170.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.74-4.08
(total 27H, series of m), 6.73 (1H, d, J=7.8 Hz), 6.87 (1H, s),
6.93 (1H, t, J=7.8 Hz), 7.11-7.17 (2H, m), 7.79 (1H, d, J=7.8 Hz),
8.00 (1H, dd, J=7.8, 2.4 Hz), 8.47 (1H, s), 8.56 (1H, s); MS (FAB)
524 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.37N.sub.5O.sub.5.HCl.H.sub.2O: C, 58.17; H, 6.97; N,
12.11. Found: C, 58.26; H, 7.26; N, 11.53.
Example 46
[0487] ##STR1671##
[0488] To a suspension of 4-aminophenylacetic acid (10 g, 66 mmol)
in 1:1 CH.sub.2Cl.sub.2:acetone (100 mL) was added o-tolyisocyanate
(8.8 g, 66 mmol). The mixture was heated to reflux for 4 hr at
which time a white precipitate had formed. The precipitate was
filtered and the solid washed generously with 1:1
CH.sub.2Cl.sub.2:acetone. The solid was recrystallized with hot
methanol and dried under vacuum to yield 14.1 g (75% yield) of the
desired 4-(o-tolylureido)phenylacetic acid 50.
Example 47
[0489] ##STR1672##
[0490] To a suspension of 2-amino-4-thiazoleacetic acid (4 g, 25
mmol) in 1:1 CH.sub.2Cl:acetone (100 mL) was added
o-tolylisocyanate (3.5 g, 26 mmol). The mixture was heated to
reflux for 8 hr at which time a yellow precipitate had formed. the
precipitate was filtered and the solid washed generously with 1:1
CH.sub.2Cl.sub.2:acetone. The solid was recrystallized with hot
methanol and dried under vacuum to yield 4.8 g (66% yield) of the
desired 2-(o-tolylureido)-4-thiazoleacetic acid 51.
Example 48
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
methylamino]benzoic acid
[0491] ##STR1673##
[0492] A stirred mixture of methyl 4-aminobenzoate (1.52 g, 10.04
mmol) and 1-tert-butoxy carbonyl prolinal (3.00 g, 15.06 mmol) in
toluene (30 mL) was heated under reflux for 3 hr. After cooling to
room temp, the solvent was evaporated in vacuo. The solid was
dissolved in MeOH (27 mL) and AcOH (3 mL), then NaBH.sub.3CN (1.33
g, 20.08 mmol) was added to the mixture, and the resulting mixture
was stirred overnight at room temp. The reaction mixture was
quenched with water, and the solvent was removed under a reduced
pressure. Water was added to the residue, and extracted with EtOAc.
The extract was washed with H.sub.2O, brine, and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure,
and the residue was chromatographed on silica-gel with
n-hexane-EtOAc (3:1, v/v) as eluent to afford 2.17 g (65%)
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethylamino]benzoate as a
pale yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 1.48 (s,
9H), 1.51-2.09 (m, 4H), 3.05-3.07 and 3.43-3.48 (br m, 1H), 3.18
(br s, 1H), 3.36 (br s, 2H), 3.84 (s, 1H), 4.06-4.08, 4.20-4.24 (br
m each, 1H), 6.49-6.65 (m, 2H), 7.84 (d, J=8.3 Hz, 2H); MS (FAB)
m/z 335 (M.sup.++1).
[0493] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethylamino]benzoate (2.17
g, 6.490 mmol) in CH.sub.2Cl.sub.2 (44 mL) was added TFA (8.7 mL)
at 0.degree. C., and the resulting mixture was stirred overnight at
room temp. The solvent was removed under a reduced pressure and the
residue was treated with 1N NaOH, The mixture was extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and the solvent was concentrated under a reduced
pressure to afford 1.34 g (88%) methyl
4-(2-pyrrolidinylmethylamino)benzoate as a brown oil, which is used
to the subsequent reaction without further purification.
[0494] The mixture of the above methyl
4-(2-pyrrolidinylmethylamino)benzoate (397.8 mg, 1.69 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (587.1 mg,
1.87 mmol), EDC (HCl) (486 mg, 2.54 mmol), HOBt (23 mg, 0.17 mmol),
and DMAP (21 mg, 0.17 mmol) in DMF (4 mL) was stirred overnight at
room temp. The mixture was diluted with EtOAc, washed with brine,
and dried over Na.sub.2SO.sub.4. The solvent was removed under a
reduced pressure. The residue was chromatographed on silica-gel
with CHCl.sub.3-MeOH (50:1, v/v) as eluent to afford 882 mg (98%)
methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl
methylamino]benzoate as a brown amorphous solid, which is used to
the subsequent reaction without further purification.
[0495] To a stirred solution of the above methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
methylamino]benzoate (882 mg, 1.662 mmol) in THF (18 mL) and MeOH
(5.0 mL) was added 1N NaOH (5.0 mL, 5.000 mmol), and the mixture
was heated under reflux for 3 days. The mixture was concentrated.
The residue was treated with 1N HCl and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated in vacuo. The solid was
recrystallized from n-hexane-diisopropyl ether --CHCl.sub.3-MeOH to
afford 180.5 mg (21%) 52 as a pale yellow amorphous solid. IR (KBr)
1604, 1535, 1511, 1454, 1255, 1224, 1174 cm.sup.-1; .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 1.79-1.99 (br m, 4H), 2.25 (s, 3H),
2.90-2.94 (m, 1H), 3.35-3.62 (m, 6H), 3.87 (s, 3H), 4.12-4.15 (br
s, 1H), 6.63-6.78 (m, 4H), 6.89-6.95 and 7.11-7.17 (m each, 3H),
7.65 (d, J=8.3 Hz, 2H), 7.80 (d, J=8.3 Hz, 1H), 8.02 (d, J=8.3 Hz,
1H), 8.47 (s, 1H), 8.57 (s, 1H), 12.0 (br s, 1H); MS (FAB) m/z 517
(M.sup.++1); Anal. Calcd for
C.sub.29H.sub.32N.sub.4O.sub.5.1H.sub.2O: C, 65.15; H, 6.41; N,
10.48. Found: C, 65.45; H, 6.33; N, 10.02.
Example 49
4-[N-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
ylmethyl]-N-methylamino]benzoic acid
[0496] ##STR1674##
[0497] To a mixture of methyl
4-[N-(2-pyrrolidinyl)methylamino]benzoate (600 mg, 1.794 mmol),
37%-formaldehyde (1.79 mL, 23.32 mmol), and NaBH.sub.3CN (368 mg,
5.561 mmol) in CH.sub.3CN (6.0 mL) was added dropwise AcOH (0.205
mL, 3.588 mmol), and the resulting mixture was stirred for 2 hr at
room temp. The reaction mixture was quenched by the addition of
sat. NaHCO.sub.3, and extracted with EtOAc. The extract was washed
with brine and dried over Na.sub.2SO.sub.4. The solvent was removed
under a reduced pressure, and the residue was chromatographed on
silica-gel with n-hexane-EtOAc (3:1, v/v) as eluent to afford 645
mg (100%) methyl
4-[N-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethyl]-N-methylamino]benzoat-
e as a colorless oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.1.50
(s, 9H), 1.76-1.91 (m, 4H), 3.07 (s, 3H), 3.15-3.43 (m, 3H),
3.67-3.71 (m, 1H), 3.85 (s, 3H), 4.11-4.17 (m, 1H), 4.37 (s, 1H),
6.75 (d, J=8.3 Hz, 2H), 7.89 (d, J=8.8 Hz, 2H); MS (FAB) m/z 349
(M.sup.++1).
[0498] To a stirred solution of methyl
4-[N-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethyl]-N-methylamino]benzoat-
e (645 mg, 1.794 mmol) in CH.sub.2Cl.sub.2 (6.5 mL) was added TFA
(1.3 mL) at 0.degree. C., and the mixture was stirred overnight at
room temp. The solvent was removed under a reduced pressure and the
residue was treated with 1N NaOH solution. The mixture was
extracted with CH.sub.2Cl.sub.2. The extract was washed with brine,
dried over Na.sub.2SO.sub.4, and the solvent was concentrated under
a reduced pressure to afford 363.2 mg (82%) of methyl
4-[N-(2-pyrrolidinyl)methyl-N-methyl]aminobenzoate as a yellowish
oil, which is used to the subsequent reaction without further
purification.
[0499] The mixture of methyl
4-[N-(2-pyrrolidinyl)methyl-N-methyl]aminobenzoate (191.8 mg, 0.772
mmol), 3-methoxy-4-(N'-2-methylphenylureido)phenylacetic acid
(258.1 mg, 0.811 mmol), EDC (hydrochloride) (221.9 mg, 1.158 mmol),
HOBt (10.0 mg, 0.077 mmol), and DMAP (9.4 mg, 0.077 mmol) in DMF
(2.0 mL) was stirred for 3 hr at room temp. The reaction mixture
was diluted with Et.sub.2O, washed with brine, and dried over
Na.sub.2SO.sub.4. The solvent was removed under a reduced pressure
to afford 482.5 mg methyl
4-[N-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nylmethyl]-N-methylamino]benzoate as a white amorphous powder,
which is used to the subsequent reaction without further
purification.
[0500] To a stirred solution of methyl
4-[N-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinylmethyl]-N-methylamino]benzoate in THF (5.0
mL) was added 1N NaOH (6.2 mL, 6.2 mmol), and the mixture was
heated under reflux for 3 days. The reaction mixture was
concentrated in vacuo. The residue was neutralized with 1N HCl, and
extracted with CH.sub.2Cl.sub.2. The extract was washed with sat.
NH.sub.4Cl, brine, dried over Na.sub.2SO.sub.4, and evaporated in
vacuo. The crude solid was recrystallized from
n-hexane-CHCl.sub.3-MeOH-isopropylether to afford 102.8 mg (25%, 2
steps) 53 as a pale yellow amorphous solid. mp 144-146; IR (KBr)
3325, 1600, 1529, 1454, 1284, 1257, 1184 cm.sup.-1; .sup.1H-NMR
(400 MHz, DMSO-d.sub.6) .delta. 1.73-1.91 (m, 3H), 2.03-2.11 (m,
1H), 3.03 (s, 3H), 3.16 (dd, J=9.3, 14.2 Hz, 1H), 3.37-3.60 (m,
4H), 3.76-3.80 (m, 1H), 3.86 (s, 3H), 4.25 (br s, 1H), 6.75 (dd,
J=1.5, 8.3 Hz, 1H), 6.86 (d, J=1.5 Hz, 1H), 6.90 (d, J=8.8 Hz, 2H),
6.95-7.01 (m, 1H), 7.12 (t, J=7.8 Hz, 1H), 7.20-7.25 (m, 1H), 7.73
(d, J=8.8 Hz, 2H), 8.01 (d, J=7.8 Hz, 1H), 8.16-8.20 (m, 1H), 8.73
(s, 1H), 9.19 (d, J=2.0 Hz, 1H), 12.0 (br s, 1H); MS (FAB) m/z 535
(M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31FN.sub.4O.sub.5.1/2H.sub.2O: C, 64.08; H, 5.93; N,
10.31; F, 3.49. Found: C, 64.17; H, 5.84; N, 10.06; F, 3.26.
Example 50
4-[N-[1-[-4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidi-
nylmethyl]-N-methylamino]-3-nitrobenzoic acid
[0501] ##STR1675##
[0502] To a mixture of methyl 4-fluoro-3-nitrobenzoate (1.58 g,
4.666 mmol) and [1-(tert-butoxycarbonyl)-2-pyrrolidinyl]methylamine
(500 mg, 2.333 mmol) in DMF (8.0 mL) was added K.sub.2CO.sub.3 (967
mg, 6.999 mmol), the resulting mixture was stirred for 3 hr at room
temp. The reaction mixture was diluted with EtOAc, washed with
water, and dried over Na.sub.2SO.sub.4. The solvent was removed
under a reduced pressure, and the residue was chromatographed on
silica-gel with n-hexane-EtOAc (3:1, v/v) as eluent to afford 834.9
mg (91%) of methyl
4-[N-[1-(tert-butoxycarbonyl)-2-pyrrolidinylmethyl]-N-methyl]amino-3-nitr-
obenzoate as a pale yellow oil, which is used to the subsequent
reaction without further purification.
[0503] To a ice-cooling solution of the above oil in
CH.sub.2Cl.sub.2 (8.3 mL) was added TFA (1.7 mL), and the reaction
mixture was stirred overnight at room temp. The solvent was removed
under a reduced pressure. The residue was treated with 1N-NaOH and
extracted with CHCl.sub.3. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and evaporated under a reduced pressure to
afford 553.6 mg (90%) methyl
4-[N-(2-pyrrolidinylmethyl)-N-methyl]amino-3-nitrobenzoate as a
pale yellow oil. .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta.
1.31-1.40 (m, 1H), 1.74-2.05 (m, 4H), 2.73-2.79 (m, 1H), 2.81-2.99
(m, 1H), 2.94 (s, 3H), 3.29-3.55 (m, 2H), 3.89 (s, 3H), 7.14 (d,
J=9.3 Hz, 1H), 7.98 (dd, J=2.0, 8.8 Hz, 1H), 8.42 (d, J=2.0 Hz,
1H); MS (FAB) m/z 294 (M.sup.++1).
[0504] A mixture of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (630.0 mg,
1.979 mmol), methyl
3-nitro-4-[N-(2-pyrrolidinyl)methyl-N-methylamino]benzoate (553.0
mg, 1.885 mmol), EDC(Hydrochloride) (542.0 mg, 2.827 mmol), HOBt
(25.5 mg, 0.189 mmol), and DMAP (23.1 mg, 0.189 mmol) in DMF (5.0
mL) was stirred at room temp. for 2 hr. The mixture was diluted
with Et.sub.2O, washed with brine, and dried over Na.sub.2SO.sub.4.
The solvent was removed under a reduced pressure, and the residue
was chromatographed on silica-gel with CHCl.sub.3-MeOH (30:1, v/v)
as eluent to afford 1.18 g (100%) methyl
4-[N-[1-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-2-pyrrolidi-
nyl methyl]-N-methylamino]-3-nitrobenzoate as a yellow foam, which
is used to the subsequent reaction without further
purification.
[0505] To a stirred solution of the above methyl
4-[N-[1-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-2-pyrrolidi-
nylmethyl]-N-methylamino]-3-nitrobenzoate (2.50 mg, 0.421 mmol) in
THF (3.0 mL) was added 1N NaOH (1.5 mL, 1.500 mmol), and the
mixture was heated under reflux overnight. After cooling, the
mixture was concentrated to a small volume. The residue was treated
with 1N HCl, and extracted with CHCl.sub.3. The extract was washed
with brine, dried over Na.sub.2SO.sub.4, and evaporated in vacuo.
The crude solid was recrystallized from n-hexane-diethyl ether
--CHCl.sub.3-MeOH to afford 194.9 mg (80%) of 54 as a yellow
amorphous solid. IR (KBr) 1685, 1610, 1529, 1454, 1284, 1259, 1228
cm.sup.-1; .sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.63-1.91
(m, 3H), 2.04-2.07 (br s, 1H), 2.60 (br s, 1H), 2.80 (s, 1H), 2.99
(s, 2H), 3.05-3.10 (m, 1H), 3.32-3.58 (m, 3H), 3.76-3.81 (m, 1H),
3.81 (s, 3H), 4.25 (br s, 1H), 6.68 (t, J=3.9 Hz, 1H), 6.71 (d,
J=8.8 Hz, 1H), 6.81-6.96 (m, 1H), 7.07 (t, J=7.3 Hz, 1H), 7.17 (dd,
J=7.8, 9.8 Hz, 1H), 7.48 (t, J=7.8 Hz, 1H), 7.85 (d, J=8.8 Hz, 1H),
7.97 (t, J=8.8 Hz, 1H), 8.11-8.20 (m, 2H), 8.68 (s, 1H), 9.14 (s,
1H), 12.8 (br s, 1H); MS (FAB) m/z 580 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.30FN.sub.5O.sub.7.1/4H.sub.2O: C, 59.63; H, 5.26; N,
11.99; F, 3.25. Found: C, 59.68; H, 5.34; N, 11.80; F, 3.21.
Example 51
3-amino-4-[N-methyl-[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxy-phenylacet-
yl]-2-pyrrolidinyl methyl]-N-methylamino]benzoic acid
[0506] ##STR1676##
[0507] A stirred solution of methyl
4-[N-[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxy-phenylacetyl]-2-pyrrolid-
inylmethyl]-N-methylamino]-3-nitrobenzoate (901.0 mg, 1.518 mmol)
in MeOH (18.0 mL) was hydrogenated over 5% Pd--C (1.35 g) at 45 psi
overnight. The mixture was filtered and the filtrate was
concentrated in vacuo. The residue was made basic with 1N NaOH
solution and extracted with CHCl.sub.3. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and the solvent was removed
under a reduced pressure. The residue was chromatographed on
silica-gel with CHCl.sub.3-MeOH (24:1, v/v) as eluent to afford
283.7 mg (48%) methyl
3-amino-4-[N-[1-[4-[N'-(2-fluorophenyl)ureido-3-methoxy-phenylacetyl]-2-p-
yrrolidinylmethyl]-N-methylamino]benzoate as a brownish amorphous
solid, which was used to the subsequent reaction without further
purification.
[0508] To a stirred solution of the above compound in THF (3.0 mL)
was added 1N NaOH solution (1.5 mL, 1.500 mmol), and the mixture
was refluxed overnight. The mixture was concentrated, treated with
1N HCl, and extracted with CHCl.sub.3. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated in vacuo. The
solid was recrystallized from n-hexane-diethyl ether
--CHCl.sub.3-MeOH to afford 179.8 mg (65%) 55 as a white amorphous
solid. IR (KBr) 1614, 1601, 1537, 1454, 1228, 1219, 1184 cm.sup.-1;
.sup.1H-NMR (400 MHz, DMSO-d.sub.6) .delta. 1.60-2.20 (m, 4H),
2.61-2.68 (m, 1H), 2.89 (s, 3H), 3.13-3.18 (m, 1H), 3.40-3.61 (m,
4H), 3.85 (s, 3H), 4.01 (br m, 1H), 4.93 (br s, 2H), 6.50-7.31 (m,
8H), 8.01 (dd, J=2.9, 8.3 Hz, 1H), 8.18 (t, J=8.3 Hz, 1H), 8.71 (s,
1H), 9.17 (d, J=1.5 Hz, 1H), 12.3 (br s, 1H); MS (FAB) m/z 550
(M.sup.++1); Anal. Calcd for
C.sub.29H.sub.32FN.sub.5O.sub.5.1/4H.sub.2O: C, 62.86; H, 5.91; N,
12.64; F, 3.43. Found: C, 62.71; H, 6.00; N, 12.39; F, 3.16.
Example 52
4-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]methylamin-
o]benzoic acid
[0509] ##STR1677##
[0510] To a stirred mixture of methyl
4-[(2-pyrrolidinyl)methylamino]benzoate (220 mg, 0.94 mmol),
4-[N'-(2-methylphenyl)ureido]phenylacetic acid (285 mg, 0.94 mmol),
4-DMAP (140 mg, 1.13 mmol) and catalytic amount of HOBT in DMF (7
ml) was added EDC.HCl (220 mg, 1.13 mmol) at room temperature. The
resulting mixture was stirred at room temperature for 20 hr. The
mixture was pored into ice-water. The solid was collected, washed
with water and air-died. The crude solid was purified by silica-gel
(20 ml) column chromatography with CHCl.sub.3-EtOAc (3:1, v/v) to
CHCl.sub.3-EtOH (9:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl]methylamino]benzoate (400 mg, 85%) as a gum.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.75-2.05 (series of m, 4H), 2.24
(s, 3H), 3.18 and 3.27 (each m, each 1H), 3.51 (m, 2H), 3.60 (s,
2H), 3.83 (s, 3H), 4.52 (m, 1H), 6.52 (m, 3H), 6.81 (s, 1H),
7.11-7.25 (series of m, 7H), 7.53 (m, 1H), 7.81 (d, J=8.8 Hz,
2H).
[0511] A mixture of methyl
4-[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl]methylami-
no]benzoate (280 mg, 0.56 mmol) in THF (3 ml) and 0.25N NaOH (6.8
ml, 1.75 mmol) was stirred for 3 hr at 60-70.degree. C. After
cooling, the mixture was poured into ice-1N HCl (3 ml). The solid
was collected, washed with water and air-dried. The crude
crystalline material was recrystallized from CHCl.sub.3-EtOH--IPE
to give 56 (180 mg, 66%) as fine needles. MW 486.56 IR (KBr) n
3367, 3294, 1712, 1606, 1539 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3-DMSO-d.sub.6) .delta. 1.80-2.05 (series of m, 4H), 2.26
(s, 3H), 2.94 (m, 1H), 3.38 and 3.56 (series of m, 3H), 3.57 (s,
2H), 4.23 (m, 2H), 6.48 (br s, 1H), 6.69 (d, J=8.8 Hz, 2H), 6.91
(t, J=7 Hz, 1H), 6.91 (m, 4H), 7.39 (d, J=8.3 Hz, 2H), 7.66 (d,
J=8.8 Hz, 2H), 7.80 (m, 2H), 8.88 (s, 1H), 11.76 (s, 1H); MS (FAB)
m/z 487 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.30N.sub.4O.sub.4.0.75.times.H.sub.2O: C, 67.24; H,
6.45; N, 11.20. Found: C, 67.13; H, 6.32; N, 11.01.
Example 53
methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-py-
rrolidinyl]methoxybenzoate
[0512] ##STR1678##
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolidi-
nyl]methoxybenzoic acid
[0513] ##STR1679##
[0514] To a stirred solution of 25-prrrolidinemethanol (15.1 g,
149.5 mmol) in dioxane (100 ml) was added a solution of
(Boc).sub.2O (32.6 g, 164.4 mmol) in dioxane (100 ml). The reaction
mixture was stirred at room temperature for 18 hr, and concentrated
in vacuo. The residue was purified by column chromatography on
silica gel with EtOAc-hexane (1:5, v/v) as eluent to give
(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methanol (31.6 g, quant.)
as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47 (s, 9H),
1.60-2.00 (m, 3H), 3.25-3.70 (4H, m), 3.92-4.00 (m, 1H).
[0515] To a stirred solution of
(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methanol (4.02 g, 20.0
mmol), methyl 4-hydroxybenzoate (3.04 g, 20.0 mmol) and Ph.sub.3P
(6.28 g, 24.0 mmol) in THF (50 ml) was added DIAD (4.85 g, 24.0
mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. The mixture was concentrated in vacuo. The
residue was purified by column chromatography on silica gel with
n-hexane-EtOAc (5:1, v/v) as eluent to give methyl
4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxybenzoate (5.4 g,
81%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47
(s, 9H), 1.88-2.04 (m, 4H), 3.41 (m, 2H), 3.91 (s, 3H), 3.90-3.92
(m, 1H), 4.11-4.16 (m, 2H), 6.94 (d, J=8.6 Hz, 2H), 7.94 (d, J=8.3
Hz, 2H).
[0516] To a stirred solution of methyl
4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxybenzoate (2.1 g,
6.27 mmol) in CH.sub.2Cl.sub.2 (9.0 ml) was added TFA (6.0 ml) at
0.degree. C. The reaction mixture was stirred at room temperature
for 0.5 hr. The mixture was concentrated in vacuo. Sat. NaHCO.sub.3
was added to the residue, and extracted with CH.sub.2Cl.sub.2. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The crude product was used to the subsequent
reaction without further purification. To a stirred solution of the
crude product (470 mg, 2.0 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (669 mg,
2.0 mmol), HOBt (405 mg, 3.0 mmol), and triethylamine (554 ml, 4.0
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (576
mg, 3.0 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (1:4, v/v) as
eluent to give 57 (900 mg, 82%) as a colorless oil. MW 552.02
.sup.1H-NMR (CDCl.sub.3) .delta. 2.04-2.10 (m, 4H), 3.51-3.70 (m,
6H), 3.87 (s, 3H), 4.11-4.18 (m, 2H), 6.77-6.88 (m, 4H), 7.23-7.34
(m, 4H), 7.91-7.96 (m, 2H), 8.17-8.19 (m, 1H).
[0517] The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to 58 (640 mg, 94%) as a white
crystalline solid. MW 537.99 mp 126-130.degree. C.; IR (KBr) 3324,
2938, 2877, 1604, 1533, 1249, 1166, 750 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.93-2.05 (m, 4H), 3.52-3.61 (m, 5H), 3.82
(s, 3H), 3.99-4.01 (m, 2H), 4.18-4.20 (m, 1H), 4.29 (m, 1H),
6.74-6.76 (d, 1H, J=8.3 Hz), 6.87 (s, 1H), 6.99-7.04 (m, 3H),
7.25-7.29 (m, 1H), 7.41-7.43 (d, 1H, J=8.1 Hz), 7.86-7.91 (m, 2H),
7.95-7.97 (m, 1H), 8.09-8.11 (d, 1H, J=8.3 Hz), 8.87-8.92 (m, 1H);
MS (FAB) m/z 538 (M.sup.++1); Anal. calcd for
C.sub.28H.sub.28N.sub.3O.sub.6.0.5H.sub.2O: C, 61.48; H, 5.34; N,
7.68; Cl, 6.48. Found: C, 61.46; H, 5.36; N, 7.62; Cl, 6.50. For Na
salt of 58: Anal. Calcd for
C.sub.28H.sub.27N.sub.3O.sub.6.Na.1.5H.sub.2O: C, 57.29; H, 5.15;
N, 7.16. Found: C, 57.48; H, 5.04; N, 6.99.
Example 54
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolidin-
yl]methoxybenzoic acid
[0518] ##STR1680##
[0519] To a stirred solution of methyl
4-2S-pyrrolidinyl)methoxybenzoate (470 mg, 2.0 mmol),
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (758 mg,
2.0 mmol), HOBt (405 mg, 3.0 mmol), and triethylamine (554 ml, 3.0
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (576
mg, 3.0 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (1:4, v/v) as
eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolidi-
nyl]methoxybenzoate (1.0 g, 84%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.04-2.10 (m, 4H), 3.52-3.54 (m, 1H), 3.62 (s,
2H), 3.70 (s, 3H), 3.88 (s, 3H), 4.13-4.19 (m, 2H), 6.79-6.94 (m,
4H), 7.20-7.31 (m, 1H), 7.91-8.12 (m, 2H), 8.13-8.15 (m, 1H).
[0520] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolidi-
nyl]methoxybenzoate (780 mg, 1.31 mmol) in THF (10.0 ml) and MeOH
(5.0 ml) was added 1N NaOH (2.0 ml, 2.0 mmol). The mixture was
stirred at 70.degree. C. for 24 hr. The mixture was concentrated in
vacuo, water was added thereto, and neutralized with 1N HCl. The
resulting solid was collected, washed with water, and dried in
vacuo to give 59 (730 mg, 96%) as a white crystalline solid. MW
582.44 mp 120-125.degree. C.; IR (KBr) 3318, 2938, 1604, 1529,
1166, 1025 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.92-1.96
(m, 4H), 3.52-3.60 (m, 5H), 3.82 (s, 3H), 3.98-4.02 (m, 1H),
4.16-4.19 (m, 1H), 4.29 (m, 1H), 6.75 (d, J=8.3 Hz, 1H), 6.87 (m,
1H), 6.94-7.04 (m, 3H), 7.29-7.33 (m, 1H), 7.57-7.59 (m, 1H),
7.85-7.96 (m, 4H), 8.72 (s, 1H), 8.91 (s, 1H); MS (FAB) m/z 582
(M.sup.++1); Anal. calcd for
C.sub.28H.sub.28N.sub.3O.sub.6Br.1.0H.sub.2O: C, 56.01; H, 5.04; N,
7.00; Br, 13.31. Found: C, 56.12; H, 4.98; N, 6.96; Br, 13.57.
Example 55
3-amino-4-[1-[4-[N'-(2-hydroxyphenyl)ureido]-3-methoxyphenylacetamido]-2-p-
yrrolidinyl methoxy]benzoic acid
[0521] ##STR1681##
[0522] To a stirred solution of 2-nitrophenol (10.0 g, 72.0 mmol)
and K.sub.2CO.sub.3 (9.96 g, 72.0 mmol) in DMF (150 mL) was added
dropwise benzyl bromide (9.40 mL, 79.2 mmol) at 0.degree. C. After
being stirred at room temperature for 3 hr, the reaction mixture
was diluted with water, which was extracted with Et.sub.2O. The
extracts were washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated to dryness. Chromatography of the residue with
hexane-EtOAc (2:1, v/v) as eluent gave 2-benzyloxy nitrobenzene
(14.7 g, 89%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
5.24 (s, 2H), 7.04 (t, J=7.8 Hz, 1H), 7.12 (d, J=7.8 Hz, 1H),
7.31-7.50 (m, 5H), 7.51 (d, J=1.5 Hz, 1H), 7.86 (dd, J=7.8, 1.5 Hz,
1H).
[0523] To a stirred solution of 2-benzyloxynitrobenzene (9.92 g,
43.3 mmol) and NiCl.sub.2 (20.3 g, 157 mmol) in MeOH (350 mL) was
added portionwise NaBH.sub.4 (8.09 g, 214 mmol) at 0.degree. C.
After disappearing of the starting material (monitored by TLC), the
mixture was evaporated off. The black precipitate was dissolved in
1N HCl, then acidic solution was alkalified by the addition of 1N
NaOH and extracted with EtOAc. The extracts were washed brine,
dried over Na.sub.2SO.sub.4, and concentrated to dryness.
Chromatography of the residue with CHCl.sub.3 as eluent gave
2-benzyloxy aniline (8.60 g, 100%) as a reddish oil. .sup.1H-NMR
(CDCl.sub.6) .delta. 3.71 (broad s, 2H), 5.06 (s, 2H), 6.68-6.86
(m, 4H), 7.32-7.44 (m, 5H); FAB-MS m/z 200 (M.sup.++1).
[0524] To a solution of 2-benzyloxyaniline (1.15 g, 5.77 mmol) in
benzene (60 mL) was added triphosgene (1.27 g, 6.35 mmol) and
Et.sub.3N (2.60 mL, 17.3 mmol) at 0.degree. C. The reaction mixture
was heated under reflux for 20 hr. The resulting mixture was
filtrated and washed with hexane, and the filtrate was concentrated
to leave a residual oil, which was chromatographed with
hexane-EtOAc (4:1, v/v) as eluent to afford tert-butyl
4-[N'-(2-benzyloxyphenyl)ureido]-3-methoxy phenylacetate (2.38 g,
89%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.44 (s,
9H), 3.44 (s, 2H), 3.78 (s, 3H), 5.07 (s, 2H), 6.73 (dd, J=8.0, 1.7
Hz, 1H), 6.78 (d, J=1.7 Hz, 1H), 6.90-6.98 (m, 3H), 7.07 (s, 1H),
7.29 (s, 1H), 7.33-7.38 (m, 5H), 7.91 (d, J=8.0 Hz, 1H), 8.14 (m,
1H).
[0525] To a solution of tert-butyl
4-[N'-(2-benzyloxyphenyl)ureido]-3-methoxyphenylacetate (2.35 g,
5.08 mmol) in CH.sub.2Cl.sub.2 (25 mL) was added TFA (25 mL) at
0.degree. C. After being stirred at room temperature. for 3 hr, the
mixture was concentrated. The residue was dissolved in 1N NaOH and
washed with Et.sub.2O. The basic water layer was poured into ice-1N
HCl and the resulting mixture was extracted with CHCl.sub.3-MeOH
(4:1, v/v). The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to dryness. The residue was
dissolved in isopropyl ether and hexane was added to this solution
until the crystallization was completed. The solid was collected to
give 4-[N'-(2-benzyloxyphenyl)ureido]-3-methoxyphenylacetic acid
(1.59 g, 77%) as a brownish solid. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 3.50 (s, 211, 3.85 (s, 3H), 5.26 (s, 2H), 6.76 (d, J=8.3
Hz, 1H), 6.83-6.89 (m, 2H), 6.91 (s, 1H), 7.01 (dd, J=8.3, 2.3 Hz,
1H), 7.31 (t, J=7.3 Hz, 1H), 7.39 (t, J=7.3 Hz, 2H), 7.49 (d, J=7.3
Hz, 2H), 7.97 (d, J=8.3 Hz, 1H), 8.04 (d, J=8.3 Hz, 1H), 8.80 (s,
1H), 8.86 (s, 1H), 12.24 (broad s, 1H); FAB-MS m/z 407
(M.sup.++1).
[0526] To a solution of
4-[N'-(2-benzyloxyphenyl)ureido]-3-methoxyphenylacetic acid (1.12
g, 2.76 mmol), methyl 4-(2-pyrrolidinylmethoxy)-3-nitrobenzoate
(890 mg, 2.76 mmol), HOBt (74.0 mg, 0.55 mmol), DMAP (67.0 mg, 0.55
mmol), and Et.sub.3N (0.58 mL, 4.13 mmol) in THF (15 mL) was added
EDC.HCl (792 mg, 4.13 mmol). After being stirred at room
temperature for 12 hr, the reaction mixture was diluted with water
and extracted with EtOAc. The extracts were washed with brine,
dried over Na.sub.2SO.sub.4, and concentrated to dryness.
Chromatography of the residue with EtOAc as eluent gave methyl
4-[1-[4-[N'-(2-benzyloxyphenyl)ureido]-3-methoxyphenylacetamido]-2-pyrrol-
idinylmethoxy]-3-nitrobenzoate (1.52 g, 82%) as a yellow amorphous
solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.91 (m, 1H), 1.95-2.17 (m,
3H), 3.47-3.53 (m, 2H), 3.56 (s, 2H), 3.60 (m, 1H), 3.68 (s, 3H),
3.90 (s, 3H), 4.11 (d, J=7.3 Hz, 1H), 4.45 (m, 1H), 5.08 (s, 2H),
6.70 (dd, J=8.3, 1.9 Hz, 1H), 6.75 (d, J=1.9 Hz, 1H), 6.91-6.99 (m,
3H), 7.16 (s, 1H), 7.18 (d, J=8.3 Hz, 1H), 7.33-7.40 (m, 6H), 7.91
(d, J=8.3 Hz, 1H), 8.11-8.16 (m, 2H), 8.46 (s, 1H).
[0527] A solution of methyl
4-[1-[4-[N'-(2-benzyloxyphenyl)ureido]-3-methoxyphenylacetamido]-2-pyrrol-
idinylmethoxy]-3-nitrobenzoate (1.52 g, 2.27 mmol) in MeOH (20 mL)
and THF (5 mL) was hydrogenated over 5% Pd--C (wet, 52.2%; 1.21 g)
under hydrogen atmosphere (4 kg/cm.sup.2) at room temperature After
being stirred for 17 hr, the catalyst was filtered off and the
filtrate was concentrated to dryness. Chromatography of the residue
with EtOAc as eluent gave methyl
3-amino-4-[1-[4-[N'-(2-hydroxyphenyl)ureido]-3-methoxyphenylacetamido]-2--
pyrrolidinylmethoxy]benzoate (1.12 g, 90%) as a brownish amorphous
solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.97-2.10 (m, 4H), 3.44 (s,
3H), 3.52-3.63 (m, 2H), 3.85 (s, 3H), 4.10-4.18 (m, 2H), 4.53 (m,
1H), 6.65-6.67 (m, 4H), 6.93-7.02 (m, 3H), 7.33 (d, J=2.2 Hz, 1H),
7.36 (dd, J=8.3, 2.2 Hz, 1H), 7.60 (s, 1H), 7.69 (d, J=8.3 Hz, 1H),
8.08 (s, 1H), 9.47 (broad s, 1H); FAB-MS m/z 549 (M.sup.++1).
[0528] To a solution of methyl
3-amino-4-[1-[4-[N'-(2-hydroxyphenyl)ureido]-3-methoxyphenyl
acetamido]-2-pyrrolidinylmethoxy]benzoate (1.12 g, 2.04 mmol) in
THF-MeOH (4:1, v/v; 20 mL) was added 1N NaOH (4.20 mL, 4.20 mmol).
After being stirred at room temperature for 24 hr, the reaction
mixture was concentrated. The residue was diluted with water and
neutralized with 1N HCl at 0.degree. C. The mixture was extracted
with CHCl.sub.3-MeOH (4:1, v/v), which was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated to dryness. Chromatography
of the residue with CHCl.sub.3:MeOH (5:1, v/v) as eluent gave 60
(352 mg, 32%) as a pale yellow amorphous solid. MW 534.56 IR (KBr)
3282, 3062, 3025, 2952, 2865, 1629, 1546, 1509, 1454, 1419
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.87-2.04 (m, 4H),
3.48-3.57 (m, 2H), 3.60 (s, 2H), 3.79 (s, 3H), 3.94 (dd, J=9.5, 7.6
Hz, 1H), 4.12 (dd, J=9.5, 3.9 Hz), 4.35 (m, 1H), 4.87 (broad s,
1H), 6.70-6.91 (m, 6H), 7.16 (dd, 1H, J=8.3, 2.0 Hz, 1H), 7.26 (d,
J=2.0 Hz, 1H), 7.96 (d, J=8.3 Hz, 1H), 7.97 (d, J=8.3 Hz, 1H), 8.80
(s, 1H), 8.82 (s, 1H); FAB-MS m/z 535 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.30N.sub.4O.sub.7.4.5H.sub.2O: C, 55.63; H, 6.39; N,
9.10. Found: C, 55.08; H, 5.06; N, 8.69.
Example 56
5-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinyl-
]methylamino]pyridine-2-carboxylic acid
[0529] ##STR1682##
5-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]methylamino]pyridine-2-carboxylic acid
[0530] ##STR1683##
[0531] To a stirred solution of
5-(methoxycarbonyl)pyridine-2-carboxylic acid (2.5 g, 13.8 mmol)
and 4-DMAP (340 mg, 2.8 mmol) in tert-BuOH (15 ml) was added
Boc.sub.2O (6 g, 27.6 mmol) at room temperature. After stirring for
2 hr at room temperature, ice and 0.2 N HCl (20 ml) was added to
the mixture and extracted with CH.sub.2Cl.sub.2. The extracts were
washed with sat. NaHCO.sub.3, dried over Na.sub.2SO.sub.4, and
evaporated. The residue was chromatographed on silica gel (50 ml)
with CH.sub.2Cl.sub.2 as eluent to give methyl
6-tert-butoxycarbonylnicotinate (2.92 g, 89%) as needles. IR (KBr)
2729, 1736, 1720, 1590, 1570 cm.sup.-1; MS (FAB) m/z 238
(M.sup.++1); Anal. Calcd for C.sub.12H.sub.15NO.sub.4: C, 60.75; H,
6.37; N, 5.90. Found: C, 60.72; H, 6.46; N, 5.78.
[0532] A mixture of methyl 6-tert-butoxycarbonylnicotinate (1.2 g,
5.06 mmol) in THF (15 ml) and 0.25 N NaOH (40 ml, 10 mol) at an
ambient temperature for 0.5 hr. The mixture was poured into ice-1N
HCl (10 ml). The solid was collected, washed with water and
air-dried. The crude solid was recrystallized from
CHCl.sub.3-EtOH--IPE to afford 6-tert-butoxycarbonylnicotinic acid
(850 mg, 76%) as needles. IR (KBr) n3095, 1728, 1705 cm.sup.-1;
.sup.1H-NMR (DMAO-d.sub.6) .delta. 1.63 (s, 9H), 8.09 (m, 1H), 8.17
(m, 1H), 8.42 (dt, J=2.4 and 8.3 Hz, 1H), 9.21 (t, J=2.4 and 8.8
Hz, 1H); MS (FAB) m/z 224 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.33N.sub.3O.sub.6: C, 36.18; H, 3.18; N, 3.84. Found:
C, 36.85; H, 3.35; N, 3.79.
[0533] To a stirred mixture of 6-tert-butoxycarbonylnicotinic acid
(1.9 g, 8.51 mmol) and triethylamine (1.17 g, 11.49 mmol) in
tert-BuOH (30 ml) and toluene (30 ml) was added a solution of
diphenyl phosphoryl azide (2.93 g, 10.64 mmol) in toluene (3 ml) at
room temperature. The resulting mixture was then heated at reflux
for 5 hr. After cooling, ice and 1N HCl (5 ml) was added to the
mixture and extracted with toluene. The extracts were washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica gel (50 ml) with toluene-EtOAc (5:1, v/v)
as eluent to give tert-butyl
5-tert-butoxycarbonylamino-2-pyridinecarboxylate (1.9 g, 76%) as a
gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.53 (s, 9H), 1.63 (s, 9H),
6.82 (br s, 1H), 8.01 (d, J=8.8 Hz, 1H), 8.17 (m, 1H), 8.46 (d,
J=2.4 Hz, 1H).
[0534] To a stirred mixture of tert-butyl
5-tert-butoxycarbonylamino-2-pyridinecarboxylate (1.9 g, 6.45 mmol)
in CH.sub.2Cl.sub.2 (20 ml) was added TFA (5 ml). The mixture was
evaporated off, and the residue was dissolved in EtOH (30 ml).
HCl-gas was induced to the solution with stirring at 0-10.degree.
C. for 10 min. The resulting stirred mixture was then heated at
reflux for 10 hr. After cooling, N.sub.2-gas was induced to remove
of large excess of HCl-gas for 15 min. the mixture was evaporated
off. The residue was alkalized by the addition of sat. NaHCO.sub.3,
and extracted with CH.sub.2Cl.sub.2. The extracts were washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica gel (30 ml) with CHCl.sub.3-EtOH (98:2,
v/v) as eluent to give ethyl 5-amino-2-pyridine carboxylate (700
mg, 65%) as a crystalline material. IR (KBr) n 3423, 3190, 1708,
1657, 15873338, 3296, 1691, 1641 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.42 (t, J=7.0 Hz, 3H), 4.11 (br s, 1H), 4.43
(q, J=7.0 Hz, 2H), 6.99 (dd, J=2.7 and 8.5 Hz, 1H), 7.95 (d, J=8.5
Hz, 1H), 8.16 (d, J=2.7 Hz, 1H); MS (FAB) m/z 167 (M.sup.+); Anal.
Calcd for C.sub.27H.sub.29ClN.sub.4O.sub.6: C, 57.47; H, 6.63; N,
16.76. Found: C, 57.27; H, 5.99; N, 16.72.
[0535] A stirred mixture of ethyl 5-amino-2-pyridinecarboxylate
(660 mg, 3.95 mmol) and 1-tert-butoxycarbonyprolinal (1.1 g, 5.33
mmol) in toluene (10 ml) was heated at reflux for 1 hr, during
which time water was azeotropically removed with Dean-Stark
water-trap. After cooling, the mixture was evaporated in vacuo. The
residue was dissolved in MeOH--AcOH (9:1, v/v, 30 ml). To the
stirred solution was added NaBH.sub.3CN (500 mg, 7.90 mmol) at
0-5.degree. C. The resulting mixture was stirred for a further 12
hr at room temperature. The mixture was stirred into ice-sat
NaHCO.sub.3 (50 ml), and extracted with CH.sub.2Cl.sub.2. The
extracts were washed with brine, dried over Na.sub.2SO.sub.4, and
evaporated. The residue was chromatographed on silica gel (50 ml)
with CHCl.sub.3-EtOAc (98:2, v/v) as eluent to give ethyl
5-[N-[2-(1-tert-butoxycarbonyl)pyrrolidinyl]methylamino]pyridine-2-carbox-
ylate (1.1 g, 70%) as a gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.38
(s, 9H), 1.42 (s, 6H), 3.93 (s, 3H), 4.29 (s, 2H), 4.67 (br s, 1H),
7.15 (d, J=8.8 Hz, 1H), 8.18 (dd, J=1.7 and 8.8 Hz, 1H), 8.52 (d,
J=1.7 Hz, 1H).
[0536] A mixture of ethyl
5-[[2-(1-tert-butoxycarbonyl)pyrrolidinylmethylamino]pyridine-2-carboxyla-
te (800 mg, 2.29 mmol) in CH.sub.2Cl.sub.2 (17 ml) and TFA (3 ml)
was stirred at room temperature for 3 hr. The mixture was
evaporated, and the residue was made basic with sat. NaHCO.sub.3.
The mixture was extracted with CH.sub.2Cl.sub.2. The extracts were
washed with brine, dried over Na.sub.2SO.sub.4--Na.sub.2CO.sub.3,
and evaporated to give ethyl
5-[(2-pyrrolidinyl)methylamino]pyridine-2-carboxylate (460 mg, 81%)
as a gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.32 (t, J=7 Hz, 3H),
1.58-2.10 (series of m, 4H), 3.12-3.28 (series of m, 3H), 3.65 (m,
1H), 4.30 (be q, J=7 Hz, 2H), 6.27 (br, 1H), 6.59 (dd, J=2.4 and
8.5 Hz, 1H), 7.65 (d, J=8.5 Hz, 1H), 7.94 (d, J=2.4 Hz, 1H).
[0537] To a stirred mixture of ethyl
5-[(2-pyrrolidinyl)methylamino]pyridine-2-carboxylate (220 mg, 0.88
mmol), 4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid
(300 mg, 0.88 mmol), 4-DMAP (135 mg, 1.10 mmol) in DMF (7 ml) was
added EDC.HCl (215 mg, 1.10 mmol) at room temperature. The
resulting mixture was stirred at room temperature for 20 hr. The
mixture was pored into ice-water. The solid was collected, washed
with water and air-died. The crude solid was purified by silica gel
(30 ml) column chromatography with CHCl.sub.3-EtOH (98:2, v/v) as
eluent and crystallized with Et.sub.2O to give
5-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrroli-
dinyl]methylamino]pyridine-2-carboxylate (420 mg, 84%) as fine
needles. IR (KBr) 3319, 1703, 1628, 1585, 1529 cm.sup.-1;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.38 (t, J=7 Hz, 3H), 1.73-2.17
(series of m, 4H), 3.19 and 3.54 (each m, each 1H), 3.63 (s, 2H),
3.70 (s, 3H), 4.39 (be q, J=7 Hz, 2H), 4.55 (m, 1H), 6.02 (br s,
1H), 6.78-6.84 (series of s and m, 3H), 6.98 (dt, J=2.4 and 8.0 Hz,
1H), 7.16 (s, 1H), 7.21-7.26 (series of m, 3H), 7.34 (dd, J=2.4 and
8.0 Hz, 1H), 7.90 (d, J=8.3 Hz, 1H), 7.98 (m, 2H), 8.16 (dd, J=1.2
and 8.8 Hz, 1H); MS (FAB) m/z 566 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.32ClN.sub.5O.sub.5.H.sub.2O: C, 59.63; H, 5.87; N,
12.37. Found: C, 60.06; H, 5.76; N, 11.95.
[0538] A mixture of ethyl
5-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidiny-
l]methylamino]pyridine-2-carboxylate (300 mg, 0.53 mmol) in
THF:MeOH (1:1, v/v, 16 ml) and 0.25N NaOH (11 ml, 2.75 mmol) was
stirred for 3 hr at room temperature. The mixture was poured into
ice-1N HCl (3 ml). The solid was collected, washed with water and
air-dried. The crude crystalline material was collected with
CH.sub.2Cl.sub.2-Et.sub.2O to give 61 (180 mg, 63%) as an amorphous
solid. MW 537.99 IR (KBr) 3319, 1701, 1620, 1585, 1533 cm.sup.-1;
.sup.1H-NMR (CDCl.sub.3) .delta. .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.80-2.05 (series of m, 4H), 2.99 (m, 1H), 3.50-3.59 (series of m,
3H), 3.60 (s, 2H), 3.86 (s, 3H), 4.11 (m, 1H), 6.78 (d, J=8.5 Hz,
1H), 6.91 (s, 1H), 6.94 (m, 1H), 7.02 (m, 1H), 7.14 (dd, J=2.5 and
8.5 Hz, 1H), 7.28 (t, J=7.0 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.78
(d, J=8.8 Hz, 1H), 7.97 (d, J=8.3 Hz, 1H), 8.08 (br s, 1H), 8.11
(m, 1H), 8.89 (s, 1H), 8.94 (s, 1H); MS (FAB) m/z 538 (M.sup.++1);
Anal. Calcd for
C.sub.27H.sub.28ClN.sub.5O.sub.5.1.5.times.H.sub.2O: C, 57.39; H,
5.53; N, 12.39. Found: C, 57.37; H, 5.54; N, 11.74.
[0539] To a stirred mixture of ethyl
5-[(2-pyrrolidinyl)methylamino]pyridine-2-carboxylate (230 mg,
0.923 mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (290 mg, 0.923 mmol), 4-DMAP (145 mg, 1.15 mmol) in DMF (7 ml)
was added EDC.HCl (225 mg, 1.15 mmol) at room temperature. The
resulting mixture was stirred at room temperature for 20 hr. The
mixture was pored into ice-water. The solid was collected, washed
with water and air-dried. The crude solid was purified by silica
gel (30 ml) column chromatography with CHCl.sub.3-EtOH (98:2, v/v)
as eluent to give ethyl
5-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methylamino]pyridine-2-carboxylate (400 mg, 80%) as fine needles.
IR (KBr) n 3325, 1709, 1618, 1585, 1531 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.39 (t, J=7 Hz, 3H), 1.73-2.07 (series of m,
4H), 2.28 (s, 3H), 3.12 and 3.49 (each m, each 1H), 3.60 (s, 2H),
4.39 (br q, J=7 Hz, 2H), 4.53 (m, 1H), 6.07 (br s, 1H), 6.23 (br s,
1H), 6.75-6.77 (series of s and m, 2H), 6.82 (dd, J=3.0 and 8.5 Hz,
1H), 7.09-7.22 (series of m, 3H), 7.49 (d, J=8.0 Hz, 1H), 7.90 (d,
J=8.5 Hz, 1H), 7.98 (d, J=2.6 Hz, 1H), 8.06 (d, J=8.8 Hz, 1H); MS
(FAB) m/z 546 (M.sup.++1); Anal. Calcd for
C.sub.30H.sub.35N.sub.5O.sub.5.1.5.times.H.sub.2O: C, 52.92; H,
6.69; N, 12.23. Found: C, 63.11; H, 6.48; N, 11.96.
[0540] A mixture of ethyl
5-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]methylamino]pyridine-2-carboxylate (D91-4596) (290 mg, 0.53 mmol)
in THF:MeOH (1:1, v/v, 16 ml) and 0.25N NaOH (11 ml, 2.75 mmol) was
stirred for 3 hr at room temperature. The mixture was poured into
ice-1N HCl (3 ml). The solid was collected, washed with water and
air-dried. The crude crystalline material was collected with
CH.sub.2Cl.sub.2-Et.sub.2O to give 62 (170 mg, 62%) as an amorphous
solid. MW 517.58 IR (KBr) 3283, 1701, 1618, 1529 cm.sup.-1;
.sup.1H-NMR (CDCl.sub.3) 6 .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.78-2.04 (series of m, 4H), 2.25 (s, 3H), 2.95-3.55 (series of m,
4H), 3.59 (s, 2H), 3.87 (s, 3H), 4.11 (m, 1H), 6.75-7.24 (series of
m, 7H), 7.83-7.97 (series of m, 3H), 8.01 (d, J=8.3 Hz, 1H), 8.13
(d, J=2.6 Hz, 1H), 8.11 (m, 1H), 8.47 (s, 1H), 8.57 (s, 1H); MS
(FAB) m/z 518 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.31N.sub.5O.sub.5.2.5.times.H.sub.2O: C, 60.50; H,
6.39; N, 12.60. Found: C, 60.31; H, 6.28; N, 12.10.
Example 57
2-[1-[[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinyl-
]methoxy]pyridine-5-carboxylic acid
[0541] ##STR1684##
[0542] To a stirred solution of methyl
2-hydroxy-5-pyridinecarboxylate (2.0 g, 13.06 mmol), PPh.sub.3 (4.2
g, 15.93 mmol) and 1-tert-butoxycarbonyl-(L)-prolinol (2.63 g,
13.06 mmol) in THF (25 ml) was added a solution of DIAD (3.3 g,
15.67 mmol) in THF (5 ml) at 0-10.degree. C. The resulting stirred
mixture was then heated at reflux for 3 hr. After cooling, the
mixture was evaporated in vacuo. The residue was chromatographed on
silica-gel (120 ml) with n-hexane-EtOAc (4:1, v/v) as eluent to
give methyl
2-[2-(1-tert-butoxycarbonyl)pyrrolidinyl]methoxypyridine-5-carboxylate
(3.0 g, 68%) as a gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.46 (s,
9H), 1.82-2.04 (series of m, 4H), 3.45 (m, 2H), 3.91 (s, 3H),
4.10-4.32 (series of m, 2H), 4.48 (m, 1H), 6.32 (br, 1H), 6.75 (d,
J=8.8 Hz, 1H), 8.15 (dd, J=2 and 8.8 Hz, 1H), 8.79 (d, J=2 Hz,
1H).
[0543] A mixture of methyl
2-[2-(1-tert-butoxycarbonyl)pyrrolidinyl]methoxypyridine-5-carboxylate
(2.9 g, 8.62 mmol) in CH.sub.2Cl.sub.2 (80 ml) and TFA (20 ml) was
stirred at room temperature for 3 hr. The mixture was evaporated,
and the residue was alkalized with sat. NaHCO.sub.3. The mixture
was extracted with CH.sub.2Cl.sub.2. The extracts were washed with
brine, dried over Na.sub.2SO.sub.4--Na.sub.2CO.sub.3, and
evaporated to give methyl
2-(2-pyrrolidinyl)methoxypyridine-5-carboxylate (1.2 g, 59%) as a
gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.58-2.050 (series of m, 4H),
2.90-3.02 (series of m, 2H), 3.87 and 3.90 (each s, 3H), 4.23 (m,
1H), 4.37 (m, 1H), 6.33 (br, 1H), 6.78 (d, J=8.5 Hz, 1H), 8.15 (dd,
J=2.2 and 8.8 Hz, 1H), 8.79 (d, J=2.2 Hz, 1H).
[0544] To a stirred mixture of methyl
2-(2-pyrrolidinyl)methoxypyridine-5-carboxylate (370 mg, 1.57
mmol), 4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid
(525 mg, 1.57 mmol), 4-DMAP (230 mg, 1.88 mmol) in DMF (10 ml) was
added EDC.HCl (360 mg, 1.88 mmol) at room temperature. The
resulting mixture was stirred at room temperature for 20 hr. The
mixture was pored into ice-water. The solid was collected, washed
with water and air-dried. The crude solid was purified by
silica-gel (30 ml) column chromatography with CHCl.sub.3-EtOH
(98:2, v/v) as eluent and crystallized with Et.sub.2O to give
methyl 2-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-2-pyrrolidinylmethoxy]pyridine-5-carboxylate (600 mg, 69%)
as an amorphous solid. .sup.1H-NMR (CDCl.sub.3) 8.21 and 2.01 (each
m, 4H), 3.45-4.50 (series of s and m, 13H which contains
amide-isomers), 6.58-8.83 (series of s and m, 12H which contains
amide-isomers)
[0545] A mixture of methyl
2-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinyl-
methoxy]pyridine-5-carboxylate (230 mg, 0.415 mmol) in THF (1 ml)
and 0.25N NaOH (4 ml, 1 mmol) was stirred for 14 hr at room
temperature and for 3 hr at 60.degree. C. After cooling, the
mixture was poured into ice-1N HCl (2 ml). The solid was collected,
washed with water and air-red. The crude crystalline material was
purified by preparative TLC plate with CHCl.sub.3-EtOH (9:1, v/v)
as eluent and crystallized with Et.sub.2O to give 63 (150 mg, 67%)
as an amorphous solid. MW 538.98 IR (KBr) 3329, 1709, 1601, 1533
cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.85-2.05 (series of m,
4H), 3.50-3.60 (series of m, 2H), 3.82 (s, 3H), 3.86 (s, 2H), 4.29
(m, 1H), 4.42 (m, 1H), 6.72-7.05 (series of m, 4H), 7.28 (m, 1H),
7.43 (d, J=8 Hz, 2H), 7.95 (d, J=8.3 Hz, 1H), 8.09 (d, J=8.3 Hz,
2H), 8.64 (m, 1H), 8.89 (s, 1H), 8.93 (s, 1H); MS (FAB) m/z 539
(M.sup.+); Anal Calcd for
C.sub.27H.sub.28ClN.sub.4O.sub.6.1.3.times.H.sub.2O: C, 57.55; H,
5.47; N, 9.94. Found: C, 57.94; H, 5.00; N, 9.45.
Example 58
5-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinylme-
thoxy]pyridine-2-carboxylic acid
[0546] ##STR1685##
[0547] To a stirred mixture of methyl
5-(2-pyrrolidinyl)methoxypyridine-2-carboxylate (370 mg, 1.57
mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (595
mg, 1.57 mmol), 4-DMAP (230 mg, 1.88 mmol) in DMF (10 ml) was added
EDC.HCl (360 mg, 1.88 mmol) at room temperature. The resulting
mixture was stirred at room temperature for 20 hr. The mixture was
pored into ice-water. The solid was collected, washed with water
and air dried. The crude solid was purified by silica-gel (30 ml)
column chromatography with CHCl.sub.3-EtOH (98:2, v/v) as eluent
and crystallized with Et.sub.2O to give methyl
5-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinylm-
ethoxypyridine-2-carboxylate (650 mg, 69%) as an amorphous solid.
IR (KBr) n 3323, 1720, 1624, 1601, 1527 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.22 and 2.00 (each m, 4H), 3.48-4.55 (series
of s and m, 13H which contains amide-isomers), 6.93-8.82 (series of
s and m, 12H which contains amide-isomers); MS (FAB) m/z 597
(M.sup.+-1) and 599 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.30BrN.sub.4O.sub.6.1.0.times.H.sub.2O: C, 54.55; H,
5.23; N, 9.09. Found: C, 54.13; H, 5.03; N, 9.33.
[0548] A mixture of methyl
5-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinylm-
ethoxy]pyridine-2-carboxylate (300 mg, 0.5 mmol) in THF:MeOH (1:1,
v/v, 2 ml) and 0.25N NaOH (4 ml, 1 mmol) was stirred for 3 hr at
room temperature and for 5 hr at 60.degree. C. After cooling, the
mixture was poured into ice-1N HCl (2 ml). The solid was collected,
washed with water and air dried. The crude crystalline material was
purified by preparative TLC plate with CHCl.sub.3-EtOH (9:1, v/v)
as eluent and crystallized with Et.sub.2O to give 64 (180 mg, 62%)
as an amorphous solid. MW 583.43 IR (KBr) n 3319, 1705, 1685, 1601,
1529 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.82-2.05
(series of m, 4H), 3.48-3.58 (series of m, 2H), 3.82 (s, 3H), 3.86
(s, 2H), 4.42-4.55 (series of m, 3H), 6.72-6.98 (series of m, 4H),
7.32 (t, J=8 Hz, 1H), 7.60 (d, J=8 Hz, 1H), 7.95 (m, 2H), 8.08 (m,
1H), 8.63 (m, 1H), 8.64 (m, 1H), 8.89 (s, 1H), 8.93 (s, 1H); MS
(FAB) m/z 583 (M.sup.+); Anal. Calcd for
C.sub.27H.sub.28BrN.sub.4O.sub.6.2.0.times.H.sub.2O: C, 52.26; H,
5.20; N, 9.03. Found: C, 52.72; H, 4.63; N, 8.50.
Example 59
4-[1-[3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetyl]-(4S)-fluoro--
(2S)-pyrrolidinyl methoxy]benzoic acid
[0549] ##STR1686##
[0550] To a stirred solution of ethyl
3-amino-2-methoxy-6-pyridylacetate (1.61 g, 7.66 mmol) in THF (10
ml) were added 2-bromophenylisocyanate (948 ml, 7.66 mmol) and
Et.sub.3N (107 ml, 0.776 mmol). After stirring overnight, the
mixture was poured into H.sub.2O (100 ml) and extracted with
CHCl.sub.3-MeOH (4:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and evaporated. The residue was
recrystallized from CHCl.sub.3-MeOH-hexane to give ethyl
3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetate (2.91 g,
93%) as a colorless crystalline powder. mp 160-163.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.19 (dt, J=7.1, 0.7 Hz, 3H),
3.69 (s, 2H), 3.95 (s, 3H), 4.07-4.13 (m, 2H), 6.90 (d, J=7.8 Hz,
1H), 6.99 (t, J=7.8 Hz, 1H), 7.33 (t, J=7.8 Hz, 1H), 7.61 (d, J=7.8
Hz, 1H), 7.96 (dd, J=7.8, 1.5 Hz, 1H), 8.31 (d, J=7.8 Hz, 1H), 8.82
(s, 1H), 9.12 (s, 1H); MS (FAB) m/z 408 (M.sup.+), 410 (M.sup.++2);
Anal. Calcd for C.sub.17H.sub.18BrN.sub.3O.sub.4.0.25H.sub.2O: C,
49.47; H, 4.52; 9.96. Found: C, 49.34; H, 4.48; N, 9.96.
[0551] A mixture of ethyl
3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetate (2.90 g,
7.10 mmol), 0.25 N NaOH (56.8 ml, 14.2 mmol), and THF (50 ml) was
stirred for 5 h. The mixture was neutralized with 1 N HCl and the
resulting precipitate was collected by filtration. The residue was
recrystallized from CHCl.sub.3-MeOH-hexane to give
3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetic acid (2.40
g, 89%) as a colorless crystalline powder. mp 195-197.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 3.59 (s, 2H), 3.95 (s, 3H), 6.88
(d, J=8.1 Hz, 1H), 6.97-7.01 (m, 1H), 7.33 (t, J=7.3 Hz, 1H), 7.61
(d, J=8.1 Hz, 1H), 7.95-7.97 (m, 1H), 8.29 (d, J=8.1 Hz, 1H), 8.81
(s, 1H), 9.10 (s, 1H), 12.35 (br s, 1H); Anal. Calcd for
C.sub.15H.sub.14BrN.sub.3O.sub.4: C, 47.39; H, 3.71; N, 11.05.
Found: C, 47.27; H, 3.59; N, 10.86.
[0552] To a stirred solution of
3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetic acid (751
mg, 1.97 mmol) and methyl
(4S)-fluoro-(2S)-pyrrolidinylmethoxybenzoate (500 mg, 1.97 mmol) in
DMF (10 ml) were added EDC.HCl (566 mg, 2.96 mmol), DMAP (cat), and
HOBt (cat.). After stirring overnight, the mixture was partitioned
between EtOAc (200 ml) and brine (200 ml). The phases were
separated. The organic phase was washed with brine (100 ml), dried
over MgSO.sub.4, and evaporated. The resulting residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[1-[3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetyl]-(4S)-fluoro-
-(2S)-pyrrolidinylmethoxy]benzoate (1.16 g, 96%) as a yellow
viscous solid.
[0553] A mixture of methyl
4-[1-[3-[N'-(2-bromophenyl)ureido]-2-methoxy-6-pyridylacetyl]-(4S)-fluoro-
-(2S)-pyrrolidinylmethoxy]benzoate (1.16 g, 1.88 mmol), 0.25 N NaOH
(15 ml, 3.75 mmol), and THF (15 ml) was stirred overnight. The
mixture was neutralized with 1 N HCl and extracted with
CHCl.sub.3-MeOH (4:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and evaporated. The resulting residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (40:1 to 10:1)
as eluent to give 65 as a pale yellow amorphous solid. MW 601.42
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.27-2.39 (m, 2H), 3.33-4.84
(series of m, 10H), 5.33-5.53 (m, 1H), 6.87-6.90 (m, 1H), 6.99 (t,
1H, J=7.6 Hz), 7.08 (d, 2H, J=9.0 Hz), 7.34 (t, 1H, J=7.6 Hz), 7.61
(d, 1H, J=7.8 Hz), 7.88 (d, 2H, J=9.0 Hz), 7.97 (d, 1H, J=8.3 Hz),
8.28-8.32 (m, 1H), 8.81-8.82 (m, 1H), 9.10-9.12 (m, 1H), 12.66 (br
s, 1H); MS (FAB) m/z 601 (M.sup.+), 603 (M.sup.++2); Anal. Calcd
for C.sub.27H.sub.26BrFN.sub.4O.sub.6: C, 53.92; H, 4.36; N, 9.32.
Found: C, 52.37; H, 4.62; N, 8.38.
Example 60
4-[(4S)-fluoro-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolid-
inylmethoxy]benzoic acid
[0554] ##STR1687##
[0555] To a stirred solution of
4-[N'-(2-methylphenyl)ureido]phenylacetic acid (337 mg, 1.18 mmol)
and methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (300
mg, 1.18 mmol) in DMF (10 ml) were added EDC.HCl (339 mg, 1.77
mmol), HOBt (cat.) and DMAP (cat.). The reaction mixture was
stirred overnight. The mixture was partitioned between EtOAc (200)
and H.sub.2O (200 ml) and the organic phase was separated. The
organic phase was washed with brine (200 ml), dried over
MgSO.sub.4, and evaporated. The residue was chromatographed on
silica gel with CHCl.sub.3-MeOH (50:1) to give methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrroli-
dinyl methoxy]benzoate (613 mg, quant) as a yellow viscous oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 2.03-2.55 (series of m, total 5H),
3.47-4.21 (series of m, total 7H), 4.44-4.60 (m, 3H), 5.21 and 5.34
(m, each, total 1H), 6.87-7.16 (m, 8H), 7.52-7.55 (m, 3H), 7.93 (d,
J=8.8 Hz, 2H), 7.99 (d, J=8.8 Hz, 1H).
[0556] To a stirred solution of methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrroli-
dinylmethoxy]benzoate (613 mg, 1.18 mmol) in THF (10 ml) was added
0.25 N NaOH (9.4 ml, 2.36 mmol). The mixture was refluxed for 1
day. After cooling to rt, the mixture was poured into 1 N HCl (50
ml) and extracted with CHCl.sub.3-MeOH (5:1, 2.times.200 ml). The
combined extracts were dried over MgSO.sub.4 and evaporated. The
residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(10:1) to give 66 (378 mg, 63%) as a colorless amorphous solid. MW
505.54 .sup.1H-NMR (DMSO-d.sub.6) .delta.0.08-2.30 (m, total 5H),
3.47-4.63 (series of m, 7H), 5.30-5.50 (m, 1H), 6.94 (t, J=7.3 Hz,
1H), 7.02-7.17 (m, 6H), 7.37-7.41 (m, 2H), 7.82-7.96 (m, 4H), 9.05
(s, 1H); MS (FAB) m/z 506 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.28FN.sub.3O.sub.5.1.75H.sub.2O: C, 62.62; H, 5.91; N,
7.82. Found: C, 62.23; H, 5.63; N, 7.18.
Example 61
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0557] ##STR1688##
[0558] To a stirred solution of
1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-proline (1.85 g, 7.93
mmol) in THF (15 ml) was added BH.sub.3-DMS (0.75 ml, 7.93 mmol) at
room temperature. After being heated at reflux for 5 h with
stirring, the mixture was cooled to room temperature and
concentrated in vacuo. The resulting residue was quenched by the
addition of H.sub.2O (100 ml) and extracted with CHCl.sub.3
(2.times.200 ml). The combined extracts were washed with brine (100
ml), dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-EtOAc (4:1) as eluent
to give
1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinylmethanol (1.76
g, quant) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.48
(s, 9H), 1.64 (m, 1H), 1.97-2.28 (m, 2H), 3.53-3.87 (series of m,
4H), 4.09-4.25 (m, 1H), 5.09 and 5.22 (m, each, total 1H).
[0559] To a stirred mixture of
1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinylmethanol (500
mg, 2.28 mmol), methyl 4-hydroxybenzoate (416 mg, 2.74 mmol),
Ph.sub.3P (719 mg, 2.74 mmol) in THF (10 ml) was added DIAD (0.54
ml, 2.74 mmol) at room temperature. The mixture was heated to
reflux for 5 h with stirring. After cooling to room temperature,
the mixture was concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-EtOAc as eluent (10:1
to 4:1) to give methyl
4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinyl
methoxy]benzoate (597 mg, 74%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.49-1.59 (m, 9H), 2.05-2.21 (m, 1H),
3.56-4.43 (series of m, 8H), 5.19 and 5.32 (m, each, total 1H),
6.97 (m, 2H), 7.98 (d, J=8.5 Hz, 2H).
[0560] A mixture of methyl
4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(590 mg, 1.67 mmol) and TFA (5 ml) in CH.sub.2Cl.sub.2 (5 ml) was
stirred for 3 h. After the mixture was concentrated in vacuo, the
residue was made basic with sat. NaHCO.sub.3 and extracted with
CHCl.sub.3 (2.times.200 ml). The combined extracts were dried over
K.sub.2CO.sub.3 and evaporated to give methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (414 mg, 98%) as a
yellow solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.89-2.02 (m, 1H),
2.16-2.31 (m, 1H), 2.98 (m, 1H), 3.35 (m, 1H), 3.46-3.68 (m, 1H),
3.86 (s, 3H), 4.00-4.08 (m, 2H), 5.16 and 5.29 (t, each, J=4.7 Hz,
total 1H), 6.91 (d, J=8.8 Hz, 2H), 7.96 (d, J=8.8 Hz, 2H).
[0561] A mixture of methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (205 mg, 0.810
mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(254 mg, 0.810 mmol), EDC.HCl (233 mg, 1.22 mmol), HOBt (cat.), and
DMAP (cat.) in DMF (10 ml) was stirred overnight. The mixture was
diluted with EtOAc (200 ml), washed with brine (2.times.100 ml),
dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2-
S)-pyrrolidinylmethoxy]benzoate (445 mg, quant) as a light brown
viscous. .sup.1H-NMR (CDCl.sub.3) .delta. 2.05-2.55 (m, total 6H),
3.55-4.13 (m, 11H), 4.48-4.60 (m, 2H), 5.20 and 5.33 (each m, total
1H), 6.29 (s, 1H), 6.79 (m, 2H), 6.96 (d, J=8.8 Hz, 2H), 7.11-7.25
(m, 3H), 7.48 (d, J=7.6 Hz, 1H), 7.93-8.09 (m, 4H).
[0562] A mixture of methyl
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido)phenylacetyl]-(2-
S)-pyrrolidinylmethoxy]benzoate (445 mg, 1.62 mmol) and 0.25 N NaOH
(15 ml, 3.75 mmol) in THF (15 ml) was stirred overnight at room
temperature then for 2 h at reflux. The mixture was acidified with
1 N HCl and extracted with CHCl.sub.3-MeOH (4:1, 2.times.200 ml).
The combined extracts were dried over MgSO.sub.4 and evaporated.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(10:1) as eluent to give 67 (260 mg, 30%) as a pale yellow
amorphous solid. MW 535.56 .sup.1H-NMR (DMSO-d.sub.6) .delta.
2.25-2.51 (m, 5H), 3.33-4.41 (series of m, 10H), 5.30-5.50 (m, 1H),
6.75-7.17 (m, 7H), 7.79 (d, J=8.1 Hz, 1H), 7.87-8.04 (m, 3H), 8.48
(m, 1H), 8.58 (m, 1H); MS (FAB) m/z 536 (M.sup.++1); Anal. Calcd
for C N.sub.3O.sub.6.H.sub.2O: C, 62.92; H, 5.83; N, 7.59. Found:
C, 62.40; H, 5.82; N, 6.93.
Example 62
4-[1-[4-[N'-(2-Bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl methoxy]benzoic acid
[0563] ##STR1689##
[0564] A mixture of methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (501 mg, 1.98
mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (750
mg, 1.98 mmol), EDC.HCl (569 mg, 2.97 mmol), HOBt (cat.) and DMAP
(cat.) in DMF (10 ml) was stirred overnight. The mixture was
diluted with EtOAc (300 ml), washed with brine (100 ml), dried over
MgSO.sub.4 and evaporated. The residue was chromatographed on
silica gel with CHCl.sub.3-EtOAc (4:1) to CHCl.sub.3-MeOH (10:1) as
eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (1.29 g,
quant) as a brown viscous oil. .sup.1H-NMR (CDCl.sub.3) .delta.
2.05-2.58 (m, 2H), 3.49-4.17 (series of m, 12H), 4.52-4.65 (m, 2H),
6.82-7.33 (series of m, 8H), 7.53 (dd, J=8.1, 1.5 Hz, 1H),
7.95-8.02 (m, 4H), 8.14 (dd, J=8.3, 1.7 Hz, 1H).
[0565] A mixture of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinylmethoxy]benzoate (1.29 g, 2.10 mmol) and 0.25 N NaOH
(17 ml, 4.20 mmol) in THF (20 ml) was refluxed for 5 h with
stirring. The mixture was poured into ice-cooled 1 N HCl (100 ml)
and extracted with CHCl.sub.3-MeOH (4:1, 2.times.200 ml). The
combined extracts were dried over MgSO.sub.4 and evaporated. The
residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(10:1) as eluent to give 68 (860 mg, 68%) as a colorless amorphous
solid. MW 600.43 .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24-2.31 (m,
2H), 3.21-4.63 (series of m, 10H), 5.31-5.51 (m, 1H), 6.74-7.10 (m,
5H), 7.32 (t, J=7.8 Hz, 1H), 7.60 (d, J=7.8 Hz, 2H), 7.87-7.99 (m,
4H), 8.74-8.75 (m, 1H), 8.92-8.94 (m, 1H); MS (FAB) m/z 601
(M.sup.++1); Anal. Calcd for
C.sub.28H.sub.27BrFN.sub.3O.sub.6.2H.sub.2O: C, 52.84; H, 4.91; N,
6.60. Found: C, 52.38; H, 4.62; N, 5.99. For Na salt of 68: mp
180-182.degree. C.; Anal. Calcd for
C.sub.28H.sub.27BrFN.sub.3NaO.sub.6.0.75H.sub.2O: C, 52.88; H,
4.36; N, 6.61. Found: C, 52.97; H, 4.36; N, 6.61.
Example 63
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0566] ##STR1690##
[0567] A mixture of methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (205 mg, 0.810
mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(254 mg, 0.810 mmol), EDC.HCl (233 mg, 1.22 mmol), HOBt (cat.) and
DMAP (cat.) in DMF (10 ml) was stirred overnight. The mixture was
diluted with EtOAc (200 ml), washed with brine (2.times.100 ml),
dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2-
S)-pyrrolidinylmethoxy]benzoate (376 mg, 81%) as a yellow foam.
.sup.1H-NMR (CDCl.sub.3) .delta.2.07-2.56 (m, 2H), 3.57-4.14
(series of m, 11H), 4.50-4.61 (m, 2H), 5.22 and 5.35 (series of m,
total 1H), 6.80-7.33 (series of m, 9H), 7.93-8.00 (m, 3H), 8.16 (d,
J=8.1 Hz, 1H).
[0568] A mixture of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2-
S)-pyrrolidinylmethoxy]benzoate (376 mg, 0.660 mmol) and 0.25 N
NaOH (15 ml, 3.75 mmol) in THF (15 ml) was stirred overnight at
room temperature then for 2 h at reflux. The mixture was acidified
with 1 N HCl and extracted with CHCl.sub.3-MeOH (4:1, 2.times.200
ml). The combined extracts were dried over MgSO.sub.4 and
evaporated. The residue was chromatographed on silica gel with
CHCl.sub.3-MeOH (20:1) as eluent to give 69 (260 mg, 30%) as a pale
yellow amorphous solid. MW 555.98 .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.24-2.501 (m, 2H), 3.48-4.65 (series of m, 10H), 5.30-5.50
(m, 1H), 6.75-7.08 (m, 5H), 7.29 (t, J=7.3 Hz, 1H), 7.43-7.45 (m,
1H), 7.89-7.98 (m, 2H), 7.99 (d, J=8.3 Hz, 1H), 8.09 (d, J=7.1 Hz,
1H), 8.90-8.96 (m, 2H); MS (FAB) m/z 556 (M.sup.++1); Anal. Calcd
for C.sub.28H.sub.27ClFN.sub.3O.sub.6.1/4H.sub.2O: C, 60.00; H,
4.95; N, 7.50. Found: C, 59.67; H, 5.08; N, 7.10.
Example 64
4-[1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-pyrrolidi-
nylmethoxy]benzoic acid
[0569] ##STR1691##
[0570] Methyl
4-[1-(4-benzyloxycarbonylaminophenylacetyl)-(4S)-fluoro-(2S)-pyrrolidinyl-
methoxy]benzoate (300 mg, 0.576 mmol) was dissolved EtOH-THF (5:1,
30 ml) and the solution was hydrogenated over 5% Pd/C (300 ml) for
12 h while stirring. The mixture was filtered to remove the
catalyst. The filtrate was concentrated under a reduced pressure.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(20:1) as eluent to give methyl
4-[1-(4-aminophenylacetyl)-(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(200 mg, 90%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
2.01-2.56 (series of m, 2H), 3.50-4.14 (series of m, 5H), 4.45-4.62
(m, 2H), 5.21 and 5.34 (each m, total 1H), 6.60-6.65 (m, 2H), 6.88
(d, J=8.8 Hz, 0.5H), 6.99-7.05 (m, 3.5H), 7.95-8.00 (m, 2H).
[0571] methyl
4-[1-(4-aminophenylacetyl)-(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(200 mg, 0.518 mmol) was dissolved THF (10 ml). Et.sub.3N (108 ul,
0.776 mmol) and 2-bromophenylisocyanate (96 ul, 0.776 mmol) were
added to the solution. The mixture was stirred overnight and
diluted with EtOAc (200 ml). The solution was washed with brine
(100 ml), dried over MgSO.sub.4, and the solvent was removed under
a reduced pressure. The residue was chromatographed on silica gel
with CHCl.sub.3-EtOAc (4:1) to CHCl.sub.3-MeOH (10:1) as eluent to
give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-pyrrolid-
inylmethoxy]benzoate (303 mg, quant) as a yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.08-2.60 (series of m, 2H), 3.56-4.69 (series
of m, 10H), 5.28 and 5.40 (m, each, total 1H), 6.84-6.92 (m, 3H),
7.03-7.10 (m, 3H), 7.14 (d, J=8.1 Hz, 1H), 7.23 (t, J=8.1 Hz, 1H),
7.39-7.44 (m, 2H), 7.89 (d, J=8.1 Hz, 1H), 7.98-8.03 (m, 2H), 8.09
(d, J=8.1 Hz, 1H).
[0572] Methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-pyrrolid-
inyl-methoxy]benzoate (300 mg, 0.513 mmol) was dissolved THF (5
ml), and 0.25 N NaOH (4.0 ml, 1.00 mmol) was added to this
solution. After being stirred for 3 days, the mixture was poured
into 1 N HCl (100 ml), and extracted with CHCl.sub.3-MeOH (5:1,
2.times.200 ml). The combined extracts were dried over MgSO.sub.4
and the solvent was removed under a reduced pressure. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) to
give 70 (209 mg, 71%) as a colorless amorphous solid. MW 570.41
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24-2.51 (m, 2H), 3.36-4.64
(series of m, 7H), 5.31-5.50 (m, 1H), 6.97 (t, J=7.8 Hz, 1H), 7.04
(d, J=8.5 Hz, 1H), 7.09 (d, J=8.8 Hz, 1H), 7.14-7.20 (m, 2H), 7.34
(t, J=7.8 Hz, 1H), 7.38-7.43 (m, 2H), 7.61 (d, J=8.1 Hz, 1H),
7.87-7.92 (m, 2H), 8.08 (d, J=8.1 Hz, 1H), 8.15 (s, 1H), 9.45-9.47
(m, 1H), 12.66 (br s, 1H); MS (FAB) m/z 572 (M.sup.++2), 570 (Mt);
Anal. Calcd for C.sub.27H.sub.25BrFN.sub.3O.sub.5.1.5H.sub.2O: C,
54.28; H, 4.72; N, 7.03. Found: C, 54.67; H, 4.51; N, 6.61.
Example 65
4-[1-[4-[N'-(2-iodophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinylmethoxy]benzoic acid
[0573] ##STR1692##
[0574] To a stirred solution of tert-butyl
4-amino-3-methoxyphenylacetate (1.94 g, 8.16 mmol) in THF (20 ml)
was added 2-iodophenylisocyanate (2.0 g, 8.16 mmol) and Et.sub.3N
(114 ul, 0.816 mmol). After stirring overnight, the mixture was
poured into 1 N HCl (200 ml). The resulting precipitate was
collected by filtration and dissolved in CHCl.sub.3 (200 ml). The
solution was dried over MgSO.sub.4 and evaporated to give
tert-butyl 4-[N'-(2-iodophenyl)ureido]-3-methoxyphenylacetate (3.93
g, quant) as a pale yellow amorphous solid. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.44 (s, 9H), 3.49 (s, 2H), 3.85 (s, 3H),
6.78-6.88 (m, 4H), 7.07 (s, 1H), 7.31-7.35 (m, 1H), 7.76 (dd,
J=7.8, 1.5 Hz, 1H), 7.95 (d, J=8.3 Hz, 1H), 7.99 (dd, J=8.3, 1.5
Hz, 1H). MS (ESI), m/z 483 (M.sup.++1).
[0575] A stirred mixture of tertbutyl
4-[N'-(2-iodophenyl)ureido]-3-methoxyphenylacetate (3.93 g, 8.16
mmol) and TFA (5 ml) in CH.sub.2Cl.sub.2 (5 ml) was refluxed for 3
h. After cooling to rt, the mixture was concentrated in vacuo and
H.sub.2O (50 ml) was added to this residue. The resulting
precipitate was collected by filtration and purified by column
chromatography on silica gel with CHCl.sub.3-MeOH (9:1) as eluent
to give 4-[N'-(2-iodophenyl)ureido]-3-methoxyphenylacetic acid
(2.89 g, 83%) as a pale yellow crystalline powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 3.62 (s, 2H), 3.88 (s, 3H), 6.78 (d, J=8.3
Hz, 1H), 6.83-6.87 (m, 1H), 6.94 (s, 1H), 7.32-7.36 (m, 1H), 7.69
(dd, J=8.3, 1.5 Hz, 1H), 7.84 (dd, J=8.3, 1.5 Hz, 1H), 7.97-8.00
(m, 1H), 8.55 (m, 1H), 8.82 (m, 1H), 12.26 (br s, 1H).
[0576] A mixture of
4-[N'-(2-iodophenyl)ureido]-3-methoxyphenylacetic acid (505 mg,
1.18 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(300 mg, 1.18 mmol), EDC.HCl (339 mg, 1.77 mmol), DMAP (catalytic
amount) and HOBt (catalytic amount) in DMF (10 ml) was stirred
overnight. The mixture was diluted with EtOAc (300 ml) and washed
with brine (2.times.200 ml). The solution was dried over MgSO.sub.4
and evaporated. The resulting residue was chromatographed on silica
gel with CHCl.sub.3-EtOAc (4:1) as eluent to give methyl
4-[1-[4-[N'-(2-iodophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinylmethoxybenzoate (500 mg, 64%) as a colorless viscous
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 07-2.58 (m, 2H), 3.59-4.20
(m, 1H), 4.51-4.64 (m, 2H), 5.24 and 5.37 (m, each, total 1H),
6.80-6.91 (m, 5H), 6.98 (d, J=8.8 Hz, 2H), 7.34 (t, J=7.8 Hz, 1H),
7.78 (dd, J=7.8, 1.2 Hz, 1H), 7.95-8.02 (m, 4H).
[0577] To a stirred solution of methyl
4-[1-[4-[N'-(2-iodophenyl)ureido]-3-methoxyphenyl
acetyl]-(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (500 mg,
0.756 mmol) in THF (6 ml) was added 0.25 N NaOH (6 ml). The
stirring was continued overnight at room temperature then 5 h at
reflux. After cooling to rt, the solution was poured into 1 N HCl
(100 ml) and extracted with CHCl.sub.3-MeOH (4:1, 2.times.200 ml).
The combined extracts were dried over MgSO.sub.4 and evaporated.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(10:1) as eluent to give 71 (295 mg, 60%) as a colorless amorphous
solid. MW 647.43 .sup.1H-NMR (DMSO-d.sub.6) .delta. 2. 2.09-2.31
(m, 2H), 3.33-4.41 (series of m, 10H), 5.30-5.50 (m, 1H), 6.77-6.92
(m, 3H), 7.03-7.09 (m, 2H), 7.34 (t, J=8.1 Hz, 1H), 7.69 (dd,
J=8.3, 1.5 Hz, 1H), 7.83-7.99 (m, 4H), 8.54 (m, 1H), 8.82 (m, 1H);
MS (FAB) m/z 648 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.27FIN.sub.3O.sub.4: C, 51.94; H, 4.20; N, 6.49.
Found: C, 51.17; H, 4.53; N, 5.76.
Example 66
4-[(4S)-fluoro-1-[4-(N'-phenylureido)phenylacetyl]-(2S)-pyrrolidinylmethox-
y]benzoic acid
[0578] ##STR1693##
[0579] To a stirred solution of ethyl 4-aminophenylacetate (6.43 g,
35.9 mmol) and Et.sub.3N (5.50 ml, 39.5 mmol) in THF (70 ml) was
added phenyl isocyanate (3.90 ml, 35.9 mmol), and the reaction
mixture was stirred at room temperature for 4 days. Resulting
precipitate was collected under a reduced pressure and the filtrate
was washed with n-hexane to give ethyl
4-(N'-phenylureido)phenylacetate (9.64 g, 90%) as a white
crystalline powder. mp 153-155.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.26 (t, J=7.1 Hz, 3H), 3.52 (s, 2H), 4.15 (q, J=7.1 Hz,
2H), 6.98-7.04 (m, 1H), 7.07-7.11 (m, 4H), 7.18-7.25 (m, 5H), 7.42
(s, 1H); MS (FAB) m/z 299 (M.sup.++1); Anal. Calcd for
C.sub.17H.sub.18N.sub.2O.sub.3: C, 68.44; H, 6.08; N, 9.39. Found:
C, 68.22; H, 6.10; N, 9.36.
[0580] To a stirred solution of ethyl
4-(N'-phenylureido)phenylacetate (9.64 g, 32.3 mmol) in THF (80 ml)
was added 0.5 N NaOH (80 ml) and the reaction mixture was heated
under reflux for 5 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl. The resulting precipitate was
collected under a reduced pressure and the crude solid was
recrystallized from MeOH--CHCl.sub.3 to give
4-(N'-phenylureido)phenylacetic acid (8.14 g, 93%) as a white
crystalline powder. MS (FAB) m/z 271 (M.sup.++1); Anal. Calcd for
C.sub.15H.sub.14N.sub.2O.sub.3: C, 66.66; H, 5.22; N, 10.36. Found:
C, 66.45; H, 5.22; N, 10.30.
[0581] A mixture of 4-(N'-phenylureido)phenylacetic acid (310 mg,
1.15 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(287 mg, 1.13 mmol), EDC.HCl (260 mg, 1.36 mmol), HOBt (185 mg,
1.37 mmol), and Et.sub.3N (190 ml, 1.36 mmol) in DMF (5 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[(4S)-fluoro-1-[4-(N'-phenylureido)phenylacetyl]-(2S)-pyrrolidinylmetho-
xy]benzoate (570 mg, 99%) as a pale yellow foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.07-2.58 (m, 2H), 3.55-3.56 (m, 1H),
3.69-3.98 (series of s and m, total 6H), 4.01-4.08 and 4.21-4.25
(each m, 1H), 4.46-4.65 (m, 2H), 5.23-5.25 and 5.38 (each m, 1H),
6.88-7.07 (m, 7H), 7.15-7.20 (m, 2H), 7.28-7.30 (m, 2H), 7.34 and
7.40 (each s, 1H), 7.71 and 7.81 (each s, 1H), 7.91-7.95 and
7.99-8.01 (each m, 2H); MS (ESI) m/z 506 (M.sup.++1).
[0582] To a stirred solution of methyl
4-[(4S)-fluoro-1-[4-(N'-phenylureido)phenylacetyl]-(2S)-pyrrolidinylmetho-
xy]benzoate (570 mg, 1.13 mmol) in THF (5 ml) was added 0.5 N NaOH
(5 ml), and the reaction mixture was heated under reflux for 5 hr.
After cooled to room temperature, the mixture was poured into ice-1
N HCl, and the resulting precipitate was collected under a reduced
pressure. The crude solid was recrystallized from
MeOH--CHCl.sub.3--IPE to give 72 (348 mg, 63%) as a white
crystalline powder. MW 491.51 mp 169-171.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.24-2.36 (m, 2H), 3.47-4.08 (m, 5H),
4.20-4.64 (m, 2H), 5.31-5.50 (m, 1H), 6.94-7.46 (series of m, total
11H), 7.87-7.92 (m, 2H), 8.64-8.67 (m, 2H), 12.63 (br s, 1H); MS
(FAB) m/z 492 (M.sup.++1); Anal. Calcd for
C.sub.27H.sub.26FN.sub.3O.sub.6.1/4H.sub.2O: C, 65.38; H, 5.38; N,
8.47; F, 3.83. Found: C, 65.13; H, 5.38; N, 8.25; F, 3.78
Example 67
4-[(4S)-fluoro-1-[3-methyl-4-(N'-phenylureido)phenylacetyl]-(2S)-pyrrolidi-
nylmethoxy]benzoic acid
[0583] ##STR1694##
[0584] To a stirred solution of tert-butyl
4-amino-3-methylphenylacetate (1.20 g, 5.42 mmol) and Et.sub.3N
(830 ml, 5.95 mmol) in THF (10 ml) was added phenyl isocyanate (650
ml, 5.98 mmol) and the reaction mixture was stirred at room
temperature overnight. The reaction mixture was concentrated to a
small volume and diluted with n-hexane. Resulting precipitate was
collected under a reduced pressure and the filtrate was washed with
n-hexane to give tert-butyl
3-methyl-4-(N'-phenylureido)phenylacetate (1.12 g, 61%) as a white
crystalline powder. mp 143-145.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.47 (s, 9H), 2.09 (s, 3H), 3.47 (s, 2H), 6.44 (s, 1H),
7.01-7.07 (m, 4H), 7.16-7.27 (m, 2H), 7.30-7.33 (m, 2H), 7.45-7.47
(m, 1H).
[0585] To a stirred solution of tertbutyl
3-methyl-4-(N'-phenylureido)phenylacetate (1.12 g, 3.29 mmol) in
CH.sub.2Cl.sub.2 (10 ml) was added TFA (10 ml) and the reaction
mixture was stirred at room temperature for 4 hr. The reaction
mixture was concentrated to a small volume and poured into
ice-H.sub.2O. The resulting precipitate was collected under a
reduced pressure and the crude solid was recrystallized from
MeOH--CHCl.sub.3 to give 3-methyl-4-(N'-phenylureido)phenylacetic
acid (680 mg, 73%) as white needles. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.22 (s, 3H), 3.46 (s, 2H), 6.93-7.05 (m, 3H), 7.25-7.29
(m, 2H), 7.43-7.46 (m, 2H), 7.72-7.74 (m, 1H), 7.90 (s, 1H), 8.98
(s, 1H), 12.26 (br s, 1H); Anal. Calcd for
C.sub.16H.sub.16N.sub.2O.sub.3: C, 67.59; H, 5.67; N, 9.85. Found:
C, 67.47; H, 5.68; N, 9.73.
[0586] A mixture of 3-methyl-4-(N'-phenylureido)phenylacetic acid
(301 mg, 1.06 mmol), methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (268 mg, 1.06
mmol), EDC.HCl (243 mg, 1.27 mmol), HOBt (172 mg, 1.27 mmol), and
Et.sub.3N (180 ml, 1.29 mmol) in DMF (5 ml) was stirred at room
temperature overnight. The mixture was diluted with H.sub.2O, and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1 to
60:1, v/v) as eluent to give methyl
4-[(4S)-fluoro-1-[3-methyl-4-(N'-phenylureido)phenylacetyl]-(2S)-pyrrolid-
inylmethoxy]benzoate (550 mg, q.y.) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.79 and 1.87 (each s, 3H), 2.04-2.61 (m, 2H),
3.52-3.54 (m, 1H), 3.73-4.27 (series of s and m, total 7H),
4.47-4.67 (m, 2H), 5.26-5.27 and 5.40 (each m, 1H), 6.79-6.99 (m,
6H), 7.14-7.18 (m, 2H), 7.27-7.31 (m, 2H), 7.40-7.44 (m, 1H),
7.89-8.01 (m, 3H); MS (ESI) m/z 520 (M.sup.++1).
[0587] To a stirred solution of methyl
4-[(4S)-fluoro-1-[3-methyl-4-(N'-phenylureido)phenylacetyl]-(2S)-pyrrolid-
inylmethoxy]benzoate (550 mg, 1.06 mmol) in THF (5 ml) was added
0.5 N NaOH (5 ml), and the reaction mixture was heated under reflux
for 2 hr. After cooled to room temperature, the mixture was poured
into ice-1 N HCl, and the resulting precipitate was collected under
a reduced pressure. The crude solid was recrystallized from
MeOH--CHCl.sub.3--IPE to give 73 (226 mg, 42%) as a white
crystalline powder. MW 505.54 mp 130-135.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.18-2.30 (series of s and m, total 5H),
3.47-3.92 (series of m, total 5H), 4.03-4.63 (m, 2H), 5.31-5.50 (m,
1H), 6.94-7.10 (m, 5H), 7.26-7.30 (m, 2H), 7.45-7.47 (m, 2H),
7.70-7.75 (m, 1H), 7.87-7.92 (m, 3H), 8.96-8.98 (m, 1H), 12.63 (br
s, 1H); MS (ESI) m/z 506 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.28FN.sub.3O.sub.5.1/2H.sub.2O: C, 65.36; H, 5.68; N,
8.17; F, 3.69. Found: C, 65.61; H, 5.71; N, 7.84; F, 3.60.
Example 68
4-[(4S)-fluoro-1-[4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetyl]-(2S)-
-pyrrolidinylmethoxy]benzoic acid
[0588] ##STR1695##
[0589] To a stirred solution of tert-butyl
4-amino-3-methylphenylacetate (1.09 g, 4.93 mmol) and Et.sub.3N
(755 ml, 5.42 mmol) in THF (10 ml) was added 2-fluorophenyl
isocyanate (610 .mu.l, 5.44 mmol) and the reaction mixture was
stirred at room temperature overnight. The reaction mixture was
concentrated to a small volume and diluted with n-hexane. Resulting
precipitate was collected under a reduced pressure and the filtrate
was washed with n-hexane to give tert-butyl
4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetate (1.31 g, 74%)
as a white crystalline powder. mp 89-91.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.47 (s, 9H), 2.06 (s, 3H), 3.49 (s, 2H), 6.62
(s, 1H), 6.92-7.09 (m, 5H), 7.21 (br s, 1H), 7.49-7.51 (m, 1H),
8.10-8.15 (m, 1H); Anal. Calcd for C.sub.20H.sub.23FN.sub.2O.sub.3:
C, 67.02; H, 6.47; N, 7.82; F, 5.30. Found: C, 66.74; H, 6.35; N,
7.85; F, 5.69.
[0590] To a stirred solution of tert-butyl
4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetate (1.25 g, 3.49
mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (10 ml) and the
reaction mixture was stirred at room temperature overnight. The
reaction mixture was concentrated to a small volume and poured into
ice-H.sub.2O. The resulting precipitate was collected under a
reduced pressure and the crude solid was recrystallized from
MeOH--HCl.sub.3--IPE to give
4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetic acid (830 mg,
79%) as white needles. .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.23 (s,
3H), 3.47 (s, 2H), 6.96-7.30 (m, 5H), 7.74-7.76 (m, 1H), 8.17-8.20
(m, 1H), 8.33 (s, 1H), 8.94 (s, 1H), 12.27 (br s, 1H); Anal. Calcd
for C.sub.16H.sub.15FN.sub.2O.sub.3: C, 63.57; H, 5.00; N, 9.27; F,
6.28. Found: C, 63.28; H, 5.00; N, 9.14; F, 6.43.
[0591] A mixture of
4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetic acid (321 mg,
1.06 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(269 mg, 1.06 mmol), EDC.HCl (244 mg, 1.27 mmol), HOBt (172 mg,
1.27 mmol), and Et.sub.3N (177 ml, 1.27 mmol) in DMF (5 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1, v/v) as eluent to give methyl
4-[(4S)-fluoro-1-[4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetyl]-(2S-
)-pyrrolidinyl methoxy]benzoate (560 mg, 98%) as a white foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.78 and 1.86 (each s, 3H),
2.16-2.65 (m, 2H), 3.58-3.61 (m, 1H), 3.74-4.15 (series of s and m,
total 7H), 4.29-4.34 and 4.46-4.49 (each m, 1H), 4.64-4.73 (m, 1H),
5.29-5.34 and 5.43-5.47 (each m, 1H), 6.84-6.97 (m, 6H), 7.04-7.07
(m, 1H), 7.21 (br s, 1H), 7.55-7.59 (m, 1H), 7.85-8.02 (m, 3H),
8.18-8.22 (m, 1H); MS (ESI) m/z 538 (M.sup.++1).
[0592] To a stirred solution of methyl
4-[(4S)-fluor-1-[4-[N'-(2-fluorophenyl)ureido]-3-methylphenylacetyl]-(2S)-
-pyrrolidinylmethoxy]benzoate (560 mg, 1.04 mmol) in THF (5 ml) was
added 0.5 N NaOH (5 ml), and the reaction mixture was heated under
reflux for 5 hr. After cooled to room temperature, the mixture was
poured into ice-1 N HCl, and the resulting precipitate was
collected under a reduced pressure. The crude solid was
recrystallized from MeOH--CHCl.sub.3--IPE to give 74 (297 mg, 42%)
as a white crystalline powder. MW 523.53 mp 137-143.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.20-2.31 (series of s and m,
total 5H), 3.56-3.92 (series of m, total 5H), 4.03-4.63 (m, 2H),
5.31-5.50 (m, 1H), 6.96-7.26 (series of m, total 7H), 7.72-7.77 (m,
1H), 7.87-7.92 (m, 2H), 8.17-8.22 (m, 1H), 8.32-8.36 (m, 1H),
8.94-8.95 (m, 1H), 12.66 (br s, 1H); MS (ESI) m/z 524 (M.sup.++1);
Anal. Calcd for C.sub.28H.sub.27F.sub.2N.sub.3O.sub.5: C, 64.24; H,
5.20; N, 8.03; F, 7.26. Found: C, 64.44; H, 5.75; N, 7.40; F,
6.73.
Example 69
4-[(4S)-fluoro-1-[4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetyl]-(2S)-
-pyrrolidinylmethoxy]benzoic acid
[0593] ##STR1696##
[0594] To a stirred solution of ethyl 4-aminophenylacetate (1.13 g,
6.31 mmol) and Et.sub.3N (965 ml, 6.92 mmol) in THF (10 ml) was
added 2-trifluoromethylphenyl isocyanate (953 ml, 6.31 mmol) and
the reaction mixture was stirred at room temperature for 2 days.
Resulting precipitate was collected under a reduced pressure and
the filtrate was washed with n-hexane to give ethyl
4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetate (1.93 g, 84%)
as white needles. mp 137-139.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.25-1.29 (m, 3H), 3.59 (s, 2H), 4.15-4.20 (m, 2H), 7.05
(br s, 1H), 7.13-7.23 (m, 6H), 7.47-7.51 (m, 1H), 7.54-7.56 (m,
1H), 8.01-8.03 (m, 1H).
[0595] To a stirred solution of ethyl
4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetate (1.93 g, 5.27
mmol) in THF (10 ml) was added 1 N NaOH (10 ml) and the reaction
mixture was heated under reflux for 5 hr. After cooled to room
temperature, the mixture was poured into ice-1 N HCl. The resulting
precipitate was collected under a reduced pressure and the crude
solid was recrystallized from MeOH--CHCl.sub.3--IPE to give
4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetic acid (910 mg,
51%) as a white crystalline powder. mp 224-225.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 3.50 (s, 2H), 7.18 (d, J=8.3 Hz,
2H), 7.25-7.29 (m, 1H), 7.40 (d, J=8.3 Hz, 2H), 7.62-7.69 (m, 2H),
7.95-7.97 (m, 1H), 8.06 (s, 1H), 9.37 (s, 1H), 12.27 (br s, 1H);
Anal. Calcd for C.sub.16H.sub.13F.sub.3N.sub.2O.sub.3: C, 56.81; H,
3.87; N, 8.28; F, 16.85. Found: C, 56.68; H, 3.87; N, 8.16; F,
16.89.
[0596] A mixture of
4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetic acid (302 mg,
0.89 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(226 mg, 0.89 mmol), EDC.HCl (205 mg, 1.07 mmol), HOBt (145 mg,
1.07 mmol), and Et.sub.3N (150 ml, 1.08 mmol) in DMF (5 ml) was
stirred at room temperature for 3 days. The mixture was diluted
with H.sub.2O, and extracted with EtOAc. The extract was washed
with brine, dried over Na.sub.2SO.sub.4, and evaporated. The
residue was purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 60:1, v/v) as eluent to give methyl
4-[(4S)-fluoro-1-[4-[N'-(2-trifluoromethylphenyl)ureido]phenylacet-
yl]-(2S)-pyrrolidinylmethoxy]benzoate (463 mg, 90%) as a pale
yellow foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.09-2.60 (m, 2H),
3.56-4.12 (series of s and m, total 8H), 4.26-4.65 (m, 2H),
5.26-5.29 and 5.39-5.42 (each m, total 1H), 6.87-6.93 (m, 2H),
6.99-7.13 (m, 5H), 7.30-7.33 (m, 1H), 7.44-7.53 (m, 2H), 7.88-7.92
(m, 1H), 7.99-8.04 (m, 2H), 8.09-8.15 (m, 1H); MS (ESI) m/z 574
(M.sup.++1).
[0597] To a stirred solution of methyl
4-[(4S)-fluoro-1-[4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetyl]-(2S-
)-pyrrolidinylmethoxy]benzoate (460 mg, 0.80 mmol) in THF (5 ml)
was added 0.5 N NaOH (5 ml), and the reaction mixture was heated
under reflux for 5 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl, and the resulting precipitate
was collected under a reduced pressure. The crude solid was
recrystallized from MeOH--CHCl.sub.3--IPE to give 75 (169 mg, 38%)
as a white crystalline powder. MW 559.51 mp 130-135.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24-2.30 (m, 2H), 3.51-4.24
(series of m, total 5H), 4.38-4.40 and 4.61 (each m, total 2H),
5.31-5.50 (m, 1H), 7.03-7.42 (series of m, total 7H), 7.62-7.69 (m,
2H), 7.87-8.07 (m, 4H), 9.36-9.37 (m, 1H), 12.64 (br s, 1H); MS
(ESI) m/z 560 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.23F.sub.4N.sub.3O.sub.5: C, 60.11; H, 4.50; N, 7.51;
F, 13.58. Found: C, 60.10; H, 4.85; N, 7.01; F, 12.90.
Example 70
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-trifluoromethylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinyl methoxy]benzoic acid
[0598] ##STR1697##
[0599] To a stirred solution of tert-butyl
4-amino-3-methoxyphenylacetate (1.11 g, 4.68 mmol) and Et.sub.3N
(720 ml, 5.17 mmol) in THF (10 ml) was added
2-trifluoromethylphenyl isocyanate (707 ml, 4.68 mmol) and the
reaction mixture was stirred at room temperature for 2 days. The
reaction mixture was concentrated to a small volume and diluted
with n-hexane. Resulting precipitate was collected under a reduced
pressure and washed with n-hexane to give tertbutyl
3-methoxy-4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetate (1.11
g, 56%) as a white crystalline powder. mp 131-133.degree. C.;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.44 (s, 9H), 3.49 (s, 2H), 3.85
(s, 3H), 6.83-6.88 (m, 3H), 6.98 (br s, 1H), 7.17-7.21 (m, 1H),
7.52-7.59 (m, 2H), 7.89-7.91 (m, 1H), 8.04-8.06 (m, 1H).
[0600] To a stirred solution of tert-butyl
3-methoxy-4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetate
(10.11 g, 2.62 mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (10
ml), and the reaction mixture was stirred at room temperature for 4
hr. The reaction mixture was concentrated to a small volume and
poured into ice-H.sub.2O. The resulting precipitate was collected
under a reduced pressure and the crude solid was recrystallized
from MeOH--CHCl.sub.3--IPE to give
3-methoxy-4-[N-(2-trifluoromethylphenyl)ureido]phenylacetic acid
(839 mg, 87%) as a white crystalline powder. mp 218-220.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 3.51 (s, 2H), 3.87 (s, 3H),
6.76-6.79 (m, 1H), 6.93-6.94 (m, 1H), 7.27-7.30 (m, 1H), 7.61-7.69
(m, 2H), 7.82-7.84 (m, 1H), 7.97-7.99 (m, 1H), 8.71 (s, 1H), 8.89
(s, 1H), 12.30 (br s, 1H); Anal. Calcd for
C.sub.17H.sub.15FN.sub.2O.sub.4: C, 55.44; H, 4.11; N, 7.61; F,
15.47. Found: C, 55.30; H, 4.08; N, 7.63; F, 15.13.
[0601] A mixture of
3-methoxy]-4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetic acid
(353 mg, 0.96 mmol), methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (243 mg, 0.96
mmol), EDC.HCl (221 mg, 1.15 mmol), HOBt (156 mg, 1.15 mmol), and
Et.sub.3N (160 ml, 1.15 mmol) in DMF (5 ml) was stirred at room
temperature overnight. The mixture was diluted with H.sub.2O, and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1 to
60:1, v/v) as eluent to give methyl
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-trifluoromethylphenyl)ureido]phenyla-
cetyl]-(2S)-pyrrolidinylmethoxy]benzoate (570 mg, 98%) as a white
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.05-2.58 (m, 2H), 3.56-4.21
(series of m, total 11H), 4.05-4.64 (m, 2H), 5.23-5.25 and
5.36-5.37 (each m, total 1H), 6.79-6.82 (m, 2H), 6.89-7.00 (m, 2H),
7.16-7.20 (m, 2H), 7.39-7.43 (m, 1H), 7.51-7.59 (m, 2H), 7.93-8.02
(m, 4H); MS (ESI) m/z 604 (M.sup.++1).
[0602] To a stirred solution of methyl
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-trifluoromethyl
phenyl)ureido]phenylacetyl]-(2S)-pyrrolidinylmethoxy]benzoate (570
mg, 0.94 mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml), and the
reaction mixture was heated under reflux for 2 hr. After cooled to
room temperature, the mixture was poured into ice-1 N HCl, and the
resulting precipitate was collected under a reduced pressure. The
crude solid was recrystallized from MeOH--CHCl.sub.3--IPE to give
76 (234 mg, 42%) as a white crystalline powder. MW 589.54 mp
129-132.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.23-2.29
(m, 2H), 3.54-4.38 (series of s and m, total 8H), 4.40-4.61 (m,
2H), 5.30-5.36 and 5.43-5.49 (each m, total 1H), 6.72-6.91 (m, 2H),
7.02-7.08 (m, 2H), 7.25-7.29 (m, 1H), 7.59-7.67 (m, 2H), 7.81-7.99
(m, 4H), 8.69-8.70 (m, 1H), 8.87-8.90 (m, 1H), 12.67 (br s, 1H); MS
(ESI) m/z 589 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.27F.sub.4N.sub.3O.sub.6: C, 59.08; H, 4.62; N, 7.13.
Found: C, 59.22; H, 5.10; N, 6.58.
Example 71
4-[(4S)-fluoro-1-[3-methyl-4-[N'-(2-trifluoromethylphenyl)ureido]phenylace-
tyl]-(2S)-pyrrolidinylmethoxy]benzoic acid
[0603] ##STR1698##
[0604] To a stirred solution of tert-butyl
4-amino-3-methylphenylacetate (927 mg, 4.19 mmol) and Et.sub.3N
(645 .mu.L, 4.63 mmol) in THF (10 ml) was added
2-trifluoromethylphenyl isocyanate (633 .mu.l], 4.19 mmol) and the
reaction mixture was stirred at room temperature for 2 days. The
reaction mixture was concentrated to a small volume and diluted
with n-hexane. Resulting precipitate was collected under a reduced
pressure and the filtrate was washed with n-hexane to give
tert-butyl
3-methyl-4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetate (1.06
g, 62%) as a white crystalline powder. mp 178-180.degree. C.;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.44 (s, 9H), 2.25 (s, 3H), 3.51
(s, 2H), 6.38 (br s, 1H), 7.12-7.18 (m, 3H), 7.36-7.37 (m, 1H),
7.49-7.53 (m, 2H), 8.13-8.16 (m, 1H).
[0605] To a stirred solution of tert-butyl
3-methyl-4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetate (1.06
g, 2.60 mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (10 ml) and
the reaction mixture was stirred at room temperature for 4 hr. The
reaction mixture was concentrated to a small volume and poured into
ice-H.sub.2O. The resulting precipitate was collected under a
reduced pressure and the crude solid was recrystallized from
MeOH--CHCl.sub.3--IPE to give
3-methyl-4-[1N'-(2-trifluoromethylphenyl)ureido]phenylacetic acid
(702 mg, 77%) as a white crystalline powder. mp 262-263.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24 (s, 3H), 3.48 (s, 2H), 7.03
(d, J=8.3 Hz, 1H), 7.08 (s, 1H), 7.26-7.30 (m, 1H), 7.61-7.69 (m,
3H), 7.88 (d, J=8.3 Hz, 1H), 8.39 (s, 1H), 8.55 (s, 1H), 12.28 (br
s, 1H); Anal. Calcd for C.sub.17H.sub.15F.sub.3N.sub.2O.sub.3: C,
57.96; H, 4.29; N, 7.95; F, 16.18. Found: C, 57.73; H, 4.23; N,
7.92; F, 16.05.
[0606] A mixture of
3-methyl-4-[N'-(2-trifluoromethylphenyl)ureido]phenylacetic acid
(359 mg, 1.02 mmol), methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (258 mg, 1.02
mmol), EDC.HCl (234 mg, 1.22 mmol), HOBt (165 mg, 1.22 mmol), and
Et.sub.3N (170 .mu.l, 1.22 mmol) in DMF (5 ml) was stirred at room
temperature overnight. The mixture was diluted with H.sub.2O, and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1 to
60:1, v/v) as eluent to give methyl
4-[(4S)-fluoro-1-[3-methyl-4-[N'-(2-trifluoromethylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinylmethoxy]benzoate (612 mg, q.y.) as a white
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 1.92 and 2.00 (each s, total
3H), 2.09-2.61 (m, 2H), 3.56-4.29 (series of m, total 8H),
4.45-4.48 and 4.59-4.64 (each m, total 2H), 5.24-5.30 and 5.38-5.44
(each m, total 1H), 6.90-7.14 (m, 5H), 7.22-7.53 (m, 5H), 7.90-7.92
(m, 1H), 8.00-8.06 (m, 2H); MS (ESI) m/z 588 (M.sup.++1).
[0607] To a stirred solution of methyl
4-[(4S)-fluoro-1-[3-methyl-4-[N'-(2-trifluoromethylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinylmethoxy]benzoate (610 mg, 1.04 mmol) in THF
(5 ml) was added 0.5 N NaOH (5 ml), and the reaction mixture was
heated under reflux for 2 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl, and the resulting precipitate
was collected under a reduced pressure. The crude solid was
recrystallized from MeOH--CHCl.sub.3--IPE to give 77 (186 mg, 31%)
as a white crystalline powder. MW 573.54 mp 123-126.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.19-2.29 (series of s and m,
total 5H), 3.64-4.21 (series of m, total 5H), 4.36-4.60 (m, 2H),
5.30-5.36 and 5.43-5.49 (each m, total 1H), 6.97-7.08 (m, 4H),
7.24-7.28 (m, 1H), 7.59-7.68 (m, 3H), 7.85-7.90 (m, 3H), 8.37-8.39
(m, 1H), 8.54-8.55 (m, 1H), 12.67 (br s, 1H); MS (ESI) m/z 573
(M.sup.+).
Example 72
4-[(4S)-fluoro-1-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-
-pyrrolidinyl]methoxybenzoic acid
[0608] ##STR1699##
[0609] A mixture of
3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (250 mg,
0.84 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate
(400 mg, 1.06 mmol), EDC.HCl (242 mg, 1.26 mmol) and DMAP (154 mg,
1.26 mmol) in DMF (5 ml) was stirred at room temperature for 21 h.
The mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [50 g, CHCl.sub.3/MeOH
(50/1)], and then was purified on TLC [CHCl.sub.3/acetone (10/1)]
to give methyl
4-[(4S)-fluoro-1-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S-
)-pyrrolidinyl]methoxybenzoate (342 mg, 76%) as a colorless
amorphous solid. IR (KBr) 3356, 2951, 1716, 1651, 1604, 1537, 1252
cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 2.07 (d, J=6.6 Hz, 2H),
2.12 (s, 3H), 2.27 (m, 1H), 2.24 (s, 3H), 2.30-2.59 (m, 1H), 3.60
(d, J=5.3 Hz, 1H), 3.65-4.23 (m, 3H), 3.87 (s, 3H), 4.50-4.62 (m,
1H), 5.31 (d, J=52.4 Hz, 1H), 6.23 (d, J=11.2 Hz, 1H), 6.26 (d,
J=11.9 Hz, 1H), 6.87-7.27 (m, 8H), 7.54-7.65 (m, 3H), 7.94-8.01 (m,
2H); MS (FAB) m/z 534 (M.sup.++1); Anal. Calcd for
C.sub.30H.sub.32FN.sub.3O.sub.5.0.7H.sub.2O: C, 65.97; H, 6.16; F,
3.48; N, 7.69. Found: C, 66.04; H, 6.07; F, 3.55; N, 7.64.
[0610] To a stirred solution of methyl
4-[(4S)-fluoro-1-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S-
)-pyrrolidinyl]methoxybenzoate (227 mg, 0.425 mmol) in THF (3.4 ml)
was added 0.25 N NaOH (3.4 ml). After stirring at room temperature
for 4 days, the mixture was acidified with 1N HCl and extracted
with CHCl.sub.3-MeOH (10/1). The combined extracts were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified on preparative-TLC [CHCl.sub.3/MeOH (10/1)] to give 78
(190 mg, 86%) as a colorless amorphous solid. MW 519.56 IR (KBr)
3356, 2974, 1604, 1537, 1454, 1252 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.24 (s, 3H), 2.26 (s, 3H), 3.60 (d, J=3.7
Hz, 2H), 3.65-4.65 (m, 8H), 5.31-5.50 (m, 1H), 6.92-7.18 (m, 7H),
7.67-7.92 (m, 4H), 8.22-8.32 (m, 2H); MS (FAB) m/z 520 (M.sup.++1);
Anal. Calcd for C.sub.29H.sub.30FN.sub.3O.sub.7.1.1H.sub.2O: C,
64.58; H, 6.02; F, 3.52; N, 7.79. Found: C, 64.71; H, 5.90; F,
3.24; N, 7.51.
Example 73
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl methoxybenzoic acid
[0611] ##STR1700##
[0612] To a stirred mixture of tert-butyl
4-amino-3-methylphenylacetate (1.00 g, 4.52 mmol), 2-chlorophenyl
isocyanate (0.55 ml, 4.52 mmol) in THF (10 ml) was added Et.sub.3N
(0.13 ml, 0.90 mmol) at room temperature. After 6 h stirring, the
reaction mixture was concentrated in vacuo. The residue was
triturated by the addition of n-hexane, to give tertbutyl
4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetate (1.57 g, 93%)
as a pale yellow powder. mp 104-106.degree. C. (dec.); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.45 (s, 9H), 2.28 (s, 3H), 3.51 (s, 2H), 6.33
(br, 1H), 6.96 (t, J=7.6 Hz, 1H), 7.08 (br, 1H), 7.16-7.30 (m, 4H),
7.42 (m, 1H), 8.2 (d, J=8.1 Hz, 1H).
[0613] To a stirred solution of tert-butyl
4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetate (1.57 g, 4.19
mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (6 ml) at room
temperature. After 4 h stirring, the mixture was concentrated in
vacuo. The residue was triturated by the addition of water to give
4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetic acid (1.33 g,
100%) as a yellow powder. mp 243-245.degree. C. (dec.); .sup.1H-NMR
(CDCl.sub.3) .delta. 2.24 (s, 3H), 3.47 (s, 2H), 6.99-7.08 (m, 3H),
7.28 (t, J=7.6 Hz, 1H), 7.44 (dt, J=8.0, 2.4 Hz, 1H), 7.66 (dd,
J=8.3, 1.9 Hz, 1H), 8.13 (dd, J=6.1, 1.7 Hz, 1H), 8.61 (d, J=6.3
Hz, 2H); MS (ESI), m/z 319 (M.sup.++1), 321 (M.sup.++3); Anal.
Calcd for C.sub.16H.sub.15ClN.sub.2O.sub.3.0.7TFA: C, 59.33; H,
4.65; Cl, 10.85; N, 8.57. Found: C, 59.23; H, 4.64; Cl, 10.90; N,
8.40.
[0614] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetic acid (252 mg,
0.79 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate
(200 mg, 0.79 mmol), EDC.HCl (227 mg, 1.20 mmol) and DMAP (147 mg,
1.20 mmol) in DMF (5 ml) was stirred at room temperature for 17 h.
The mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was purified on TLC [CHCl.sub.3/acetone (10/1)], to give
methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinyl]methoxybenzoate (390 mg, 89%) as a colorless
amorphous solid. IR (KBr) 3340, 2951, 1712, 1624, 1604, 1533, 1438
cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.92-2.05 (m, 3H),
2.07-2.63 (m, 2H), 3.61 (d, 2H, J=8.8 Hz), 3.70-4.15 (m, 5H),
4.25-4.67 (m, 2H), 5.26-5.44 (m, 1H), 6.84-8.19 (m, 13H); MS (FAB)
m/z 554 (M.sup.++1), 556 (M.sup.++3).
[0615] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenyl
acetyl]-(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate (268 mg,
0.484 mmol) in THF (3.8 ml) was added 0.25 N NaOH (3.8 ml). After
stirring at room temperature for 1 days, the mixture was acidified
with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1). The
combined extracts were dried over Na.sub.2SO.sub.4 and concentrated
in vacuo. The residue was purified on TLC [CHCl.sub.3/MeOH (10/1)]
to give 79 (124 mg, 47%) as a colorless amorphous solid. MW 539.98
IR (KBr) 3346, 2976, 1709, 1685, 1604, 1533, 1439 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.20 (s, 3H, one of isomers),
2.24 (s, 3H, one of isomers), 2.30 (m, 1H), 3.60 (s, 2H), 3.71-4.62
(m, 6H), 5.30-5.50 (m, 1H), 7.01-7.09 (m, 5H), 7.28 (t, J=7.8 Hz,
1H), 7.44 (d, J=8.1 Hz, 1H), 7.66 (t, J=8.1 Hz, 1H), 7.87 (d, J=7.1
Hz, 2H), 8.13 (d, J=7.9 Hz, 1H), 8.62 (d, J=6.1 Hz, 2H); MS (FAB)
m/z 540 (M.sup.++1), 542 (M.sup.++3). For Na salt: Anal. Calcd for
C.sub.28H.sub.27ClFN.sub.3O.sub.7.Na.0.5EtOH.1.5H.sub.2O: C, 56.91;
H, 5.27; Cl, 5.79; F, 3.10; N, 6.87. Found: C, 56.60; H, 4.98; Cl,
5.88; F, 3.08; N, 6.52.
Example 74
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinyl]methoxybenzoic acid
[0616] ##STR1701## to a stirred mixture of tert-butyl
4-amino-3-methylphenylacetate (780 mg, 3.30 mmol), 2-bromo phenyl
isocyanate (0.41 ml, 3.30 mmol) in THF (7 ml) was added Et.sub.3N
(0.092 ml, 0.66 mmol) at room temperature. After 3 h stirring, the
reaction mixture was concentrated in vacuo. The residue was
triturated by the addition of n-hexane, to give tert-butyl
4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetate (1.57 g, 93%) as
a pale yellow powder. mp 138-145.degree. C. (dec.); .sup.1H-NMR
(CDCl.sub.3) .delta. 1.44 (s, 9H), 2.33 (s, 3H), 3.51 (s, 2H), 6.90
(dt, J=9.0, 1.4 Hz, 1H), 6.98 (br, 1H), 7.18-7.31 (m, 4H), 7.39
(dd, J=8.1, 2.9 Hz, 1H), 7.44 (d, J=8.0 Hz, 1H), 8.22 (d, J=8.3 Hz,
2H); Anal. Calcd for C.sub.20H.sub.22BrN.sub.2O.sub.3.0.2H.sub.2O:
C, 56.80; H, 5.58; N, 6.62. Found: C, 56.85; H, 5.42; N, 6.62.
[0617] To a stirred solution of tert-butyl
4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetate (1.27 g, 3.03
mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (5 ml) at room
temperature. After 1 h stirring, the mixture was concentrated in
vacuo. The residue was triturated by the addition of water, to give
4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetic acid (1.05 g,
95%) as a pale yellow powder. mp 245-248.degree. C. (dec.);
.sup.1H-NMR (CDCl.sub.3) .delta. 2.24 (s, 3H), 3.48 (s, 2H), 6.96
(dt, J=7.3, 1.5 Hz, 1H), 7.02 (d, J=8.3 Hz, 1H), 7.07 (s, 1H), 7.32
(t, J=8.1 Hz, 1H), 7.59-7.66 (m, 2H), 8.44 (s, 1H), 8.62 (s, 1H);
MS (ESI), m/z 363 (M.sup.++1), 365 (M.sup.++3); Anal. Calcd for
C.sub.16H.sub.15BrN.sub.2O.sub.3.0.7H.sub.2O: C, 51.13; H, 4.40;
Br, 21.26; N, 7.45. Found: C, 50.84; H, 4.62; Br, 21.72; N,
7.18.
[0618] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetic acid (287 mg,
0.79 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate
(200 mg, 0.79 mmol), EDC.HCl (228 mg, 1.20 mmol), HOBT (160 mg,
1.19 mmol) and Et.sub.3N (0.55 ml, 3.95 mmol) in DMF (5 ml) was
stirred at room temperature for 4 days. The mixture was poured into
ice water and extracted with EtOAc. The combined extracts were
washed with ice water and brine. After dried over Na.sub.2SO.sub.4,
the extracts were concentrated in vacuo. The residue was purified
on TLC [CHCl.sub.3/acetone (10/1)], to give methyl
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetyl]-4-fluor-
o-2-pyrrolidinyl]methoxybenzoate (440 mg, 93%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.90 and 1.97
(eachs, 3H, amide isomers), 2.05-2.62 (m, 2H), 3.58 (d, J=8.1 Hz,
1H), 3.77 (m, 1H), 3.86 and 3.89 (eachs, 3H, amide isomers),
3.92-4.64 (m, 5H), 5.24-5.42 (m, 1H), 6.83-7.23 (m, 6H), 7.41-7.62
(m, 4H), 7.86-8.09 (m, 3H); MS (ESI), m/z 598 (M.sup.++1), 600
(M.sup.++3).
[0619] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenyl
acetyl]-(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate (440 mg, 0.74
mmol) in THF (6.0 ml), 0.25 N NaOH (6.0 ml) was added. After
stirring at room temperature for 1 days, the mixture was acidified
with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1). The
combined extracts were dried over Na.sub.2SO.sub.4 and concentrated
in vacuo. The residue was purified on TLC [CHCl.sub.3/MeOH (10/1)]
to give 80 (229 mg, 53%) as a colorless amorphous solid. MW 584.44
IR (KBr) 3325, 2972, 1709, 1604, 1529, 1252 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.25 (s, 3H), 2.31 (m, 1H), 3.17 (s, 1H),
3.60 (d, J=4.7 Hz, 2H), 3.83-4.67 (m, 5H), 5.31-5.51 (m, 1H), 6.97
(t, J=7.3 Hz, 1H), 7.02-7.09 (m, 5H), 7.33 (t, J=8.0 Hz, 1H), 7.61
(d, J=7.8 Hz, 1H), 7.64 (d, J=8.3 Hz, 1H), 7.87 (d, J=8.3 Hz, 2H),
7.90 (d, J=8.8 Hz, 1H), 8.44-8.65 (m, 2H); MS (ESI), m/z 584
(M.sup.++1), 586 (M.sup.++3); Anal. Calcd for
C.sub.28H.sub.27BrFN.sub.3O.sub.7.0.4H.sub.2O: C, 56.84; H, 4.74;
Br, 13.51; F, 3.21; N, 7.10. Found: C, 56.91; H, 4.93; Br, 13.23;
F, 3.15; N, 6.88. For Na salt of 80: Anal. Calcd for
C.sub.28H.sub.27BrFN.sub.3O.sub.7.Na.1.8H.sub.2O: C, 52.64; H,
4.67; Br, 12.51; F, 2.97; N, 6.58. Found: C, 53.04; H, 4.67; Br,
12.95; F, 3.28; N, 6.11.
Example 75
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetyl]-(2S)-pyrrolidiny-
l]methoxybenzoic acid
[0620] ##STR1702##
[0621] To a stirred solution of methyl
4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxybenzoate (2.0 g,
5.9 mmol) in EtOH (10.0 ml) was added concd HCl (3.0 ml) at
0.degree. C. The reaction mixture was stirred at room temperature
for 4.0 hr. The mixture was concentrated in vacuo. The resulting
solid was collected and washed with EtOH-Et.sub.2O to give methyl
4-2S-pyrrolidinyl)methoxybenzoate HCl salt (1.4 g, 87%) as a white
crystalline solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.90-2.25 (m,
4H), 3.25-3.45 (m, 2H), 3.88 (s, 3H), 3.90-4.00 (m, 1H), 4.25-4.45
(m, 2H), 6.96 (d, J=8.5 Hz, 2H), 7.95 (d, J=8.5 Hz, 2H).
[0622] To a stirred solution of methyl
4-[(2S)-pyrrolidinyl]methoxybenzoate HCl salt (135 mg, 0.5 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methylphenylacetic acid (159 mg,
0.5 mmol), HOBt (68 mg, 0.5 mmol), and triethylamine (278 ml, 2.0
mmol) in THF (5.0 ml) and MeCN (5.0 ml) was added EDC.HCl (144 mg,
0.75 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (1:2, v/v) as
eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetyl]-2-pyrrolidinyl]-
methoxybenzoate (220 mg, 82%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.91 and 1.97 (each s, total 3H), 2.00-2.20
(m, 4H), 3.55-3.65 (m, 4H), 3.87 and 3.89 (each s, total 3H),
4.10-4.20 (m, 2H), 4.51 (m, 1H), 6.86-7.04 (m, 6H), 7.20-7.53 (m,
4H), 7.89-8.01 (m, 2H), 8.22 (d, J=8.3 Hz, 1H).
[0623] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methylphenylacetyl]-2S-pyrrolidinyl-
]methoxybenzoate (220 mg, 0.41 mmol) in THF (8.0 ml) and MeOH (4.0
ml) was added 1N NaOH (0.8 ml, 0.8 mmol). The mixture was stirred
at 70.degree. C. for 24 hr. The mixture was concentrated in vacuo,
water was added thereto, and neutralized with 1N HCl. The resulting
solid was collected, washed with water, and dried in vacuo to give
81 (220 mg, quant) as a white crystalline solid. MW 521.99 mp
122-124.degree. C.; IR (KBr) 3340, 1710, 1685, 1604, 1533, 1511,
1438 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.81-2.11 (m,
4H), 2.18 and 2.20 (each s, total 3H), 3.45-3.80 (m, 4H), 3.95-4.05
(m, 1H), 4.12-4.20 (m, 1H), 4.21-4.31 (m, 1H), 6.99-7.06 (m, 5H),
7.26-7.30 (m, 1H), 7.44 (d, J=7.8 Hz, 1H), 7.62-7.64 (m, 1H),
7.85-7.90 (m, 2H), 8.13 (d, J=6.8 Hz, 1H), 8.60-8.62 (m, 2H); MS
(FAB) m/z 522 M.sup.++1); Anal. calcd for
C.sub.28H.sub.28N.sub.3O.sub.5C.sub.10.2H.sub.2O: C, 63.99; H,
5.45; N, 7.99. Found: C, 63.90; H, 5.40; N, 7.72.
Example 76
4-[1-[4-[N-[2-bromophenyl)ureido]-3-methylphenylacetyl]-(2S)-pyrrolidinyl]-
methoxybenzoic acid
[0624] ##STR1703##
[0625] To a stirred solution of methyl
[(2S)-pyrrolidinyl]methoxybenzoate HCl salt (135 mg, 0.5 mmol),
4-[N'-(2-bromophenyl)uredio]-3-methylphenylacetic acid (181 mg, 0.5
mmol), HOBt (68 mg, 0.5 mmol), and triethylamine (278 ml, 2.0 mmol)
in THF (5.0 ml) and MeCN (5.0 ml) was added EDC.HCl (144 mg, 0.75
mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (1/2, v/v) as
eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetyl]-(2S)-pyrrolidiny-
l]methoxybenzoate (290 mg, quant) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.95 and 2.01 (each s, total 3H), 2.00-2.20
(m, 4H), 3.50-3.65 (m, 4H), 3.87 and 3.89 (each s, total 3H),
4.10-4.20 (m, 2H), 4.50 (m, 1H), 6.85-7.06 (m, 6H), 7.24-7.28 (m,
1H), 7.40-7.44 (m, 3H), 7.89-8.16 (m, 2H), 8.17-8.18 (m, 1H).
[0626] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetyl]-2S-pyrrolidinyl]-
methoxybenzoate (290 mg, 0.5 mmol) in THF (8.0 ml) and MeOH (4.0
ml) was added 1N NaOH (1.0 ml, 1.0 mmol). The mixture was stirred
at 70.degree. C. for 24 hr. The mixture was concentrated in vacuo,
water was added thereto, and neutralized with 1N HCl. The resulting
solid was collected, washed with water, and dried in vacuo to give
82 (240 mg, 85%) as a white crystalline solid. MW 566.44 mp
125-130.degree. C.; IR (KBr) 3340, 1604, 1529, 1434 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.80-2.10 (m, 4H), 2.18 and 2.20
(each s, total 3H), 3.45-3.80 (m, 4H), 3.95-4.05 (m, 1H), 4.15-4.20
(m, 1H), 4.25-4.30 (m, 1H), 6.94-7.06 (m, 5H), 7.30-7.34 (m, 1H),
7.59-7.62 (m, 2H), 7.85-7.90 (m, 2H), 8.01 (d, J=8.1 Hz, 1H), 8.44
(s, 1H), 8.62 (s, 1H); MS (FAB) m/z 566 (M.sup.+); Anal. calcd for
C.sub.2BH.sub.8N.sub.3O.sub.5Br.0.5H.sub.2O: C, 58.44; H, 5.08; N,
7.30. Found: C, 58.57; H, 4.99; N, 7.18.
Example 77
4-[1-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidiny-
lmethoxy]benzoic acid
[0627] ##STR1704##
[0628] A mixture of
3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (438 mg,
1.47 mmol), methyl 4-[(2S)-pyrrolidinylmethoxy]benzoate (420 mg,
1.79 mmol), EDC.HCl (410 mg, 2.14 mmol), HOBt (228 mg, 1.69 mmol)
and Et.sub.3N (240 ml, 1.72 mmol) in DMF (5 ml) was stirred at room
temperature overnight. The mixture was diluted with H.sub.2O and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (50:1 to
25:1, v/v) as eluent to give methyl
4-[1-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidin-
ylmethoxy]benzoate (760 mg, quant.) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.89 (s, 3H), 1.94-2.14 (m, 4H), 2.16 (s, 3H),
3.50-3.69 (m, 4H), 3.87 (s, 3H), 4.09-4.17 (m, 2H), 4.42-4.45 (m,
1H), 6.85-7.02 (m, 6H), 7.10-7.16 (m, 3H), 7.51-7.53 (m, 1H),
7.62-7.64 (m, 1H), 7.91-7.94 (m, 2H); MS (FAB) m/z 516
(M.sup.++1).
[0629] To a stirred solution of methyl
4-[1-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidin-
ylmethoxy]benzoate (420 mg, 0.71 mmol) in THF (7 ml) was added 0.5
N NaOH (7 ml) and the reaction mixture was heated under reflux for
2 hr. After cooled to room temperature, the mixture was poured into
ice-1 N HCl and the resulting precipitate was collected under a
reduced pressure. The crude solid was purified by recrystallization
from CHCl.sub.3--IPE to give 83 (526 mg, 69%) as a white
crystalline powder. MW 501.57 mp 191-193.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.87-2.10 (m, 4H), 2.20 (s, 3H), 2.26 (s,
3H), 3.44-3.79 (series of m, total 4H), 3.99-4.45 (series of m,
total 3H), 6.91-7.17 (series of m, total 7H), 7.66-7.68 (m, 1H),
7.80-7.90 (m, 3H), 8.19-8.21 (m, 2H), 12.62 (br s, 1H); MS (FAB)
m/z 502 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.5.1/4H.sub.2O: C, 68.83; H, 6.27; N,
8.30. Found: C, 68.81; H, 6.17; N, 8.23.
Example 78
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetyl]-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0630] ##STR1705##
[0631] To a stirred solution of tert-butyl
4-amino-3-methylphenylacetate (1.36 g, 6.15 mmol) and triethylamine
(170 ml, 1.23 mmol) in THF (30 ml) was added 2-methoxyphenyl
isocyanate (820 ml, 6.15 mmol), and the resulting mixture was
stirred for 27 h. The mixture was concentrated to a small volume in
vacuo, and hexane was added to the residue to give precipitate,
which was collected by filtration to give tert-butyl
4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetate (1.74 g, 76%)
as a white crystalline material. mp 157-158.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.46 (s, 9H), 2.30 (s, 3H), 3.50 (s, 2H), 3.76
(s, 3H), 6.43 (s, 1H), 6.83 (br d, J=8.4 Hz, 1H), 6.95 (br d, J=8.0
Hz, 1H), 6.98-6.99 (m, 2H), 7.13 (br s, 1H), 7.23 (br s, 1H), 7.48
(d, J=8.8 Hz, 1H), 8.14 (d, J=8.4 Hz, 1H); MS (ESI) m/z 371
(M.sup.++H).
[0632] To a stirred solution of tert-butyl
4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetate (1.32 g, 3.56
mmol) in CH.sub.2Cl.sub.2 (15 ml) was added trifluoroacetic acid
(10 ml), and the resulting mixture was heated under reflux for 30
min. The mixture was concentrated in vacuo and added water to give
precipitate which was collected by filtration. The crude solid was
recrystallized from EtOH/hexane to give
4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetic acid (932 mg,
83%) as a white powder. mp 260-264.degree. C.; .sup.1H-NMR
(CD.sub.3OD) .delta. 2.30 (s, 3H), 3.55 (s, 2H), 4.87 (s, 3H),
6.87-6.92 (m, 2H), 6.97-6.99 (m, 2H), 7.10-7.24 (m, 2H), 7.53-7.58
(m, 1H), 8.04 (d, J=7.2 Hz, 1H); MS (ESI) m/z 314 (M.sup.+).
[0633] To a stirred solution of
4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetic acid (336 mg,
1.07 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(271 mg, 1.07 mmol) and N,N-dimethylaminopyridine (130 mg, 1.07
mmol) in DMF (10 ml) was added EDC.HCl (226 mg, 1.18 mmol) at rt,
and the resulting mixture was stirred for 20 h. The mixture was
poured into 1N-HCl aq. and extracted with EtOAc. The organic layer
was washed with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (10:1) as eluent to give methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetyl]-(2-
S)-pyrrolidinylmethoxy]benzoate (583 mg, 99%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) mixture of rotamars,
.delta. 2.05 and 2.12 (s, total 3H), 2.05-2.61 (m, 2H), 3.55-4.73
(series of m, 13H), 4.51-4.66 (m, 2H), 5.26-5.40 (m, 1H), 6.72-7.01
(series of m, 8H), 7.38-8.13 (series of 13H); MS (ESI) m/z 550
(M.sup.++H).
[0634] To a stirred solution of methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]-3-methylphenylacetyl]-(2-
S)-pyrrolidinylmethoxy]benzoate (557 mg, 1.01 mmol) in MeOH-THF
(1:1, 10 ml) was added 1.0N-NaOH aq. (4.05 ml, 4.05 mmol) at rt,
and the resulting mixture was heated at 60.degree. C. for 2 h. The
mixture was poured into 1N-HCl aq. and extracted with EtOAc. The
organic layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give 84 (492 mg, 91%) as a colorless amorphous solid. MW 535.56
.sup.1H-NMR (CD.sub.3OD), mixture of rotamars, .delta. 2.96 (s,
3H), 2.11-2.45 (m, 2H), 3.64-4.15 (series of m, 5H), 3.91 (s, 3H),
4.41-4.45 (m, 1H), 4.52-4.61 (m, 1H), 5.25-5.38 (m, 1H), 6.84-7.10
(series of m, 7H), 7.54-7.58 (m, 1H), 7.93 (d, J=8.8 Hz, 2H), 8.02
(d, J=8.8 Hz, 2H); MS (ESI) m/z 536 (M.sup.++H), 538
(M.sup.++Na.sup.+).
Example 79
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]phenylacetyl]-(2S)-pyrroli-
dinylmethoxy]benzoic acid
[0635] ##STR1706##
[0636] To a stirred solution of ethyl 4-amino-3-methylphenylacetate
(1.32 g, 7.37 mmol) and triethylamine (205 ml, 1.47 mmol) in THF
(20 ml) was added 2-methoxyphenyl isocyanate (980 ml, 7.37 mmol),
and the resulting mixture was stirred for 23 h. The mixture was
concentrated to a small volume in vacuo and hexane was added to the
residue to give precipitate, which was collected to give ethyl
4-[N'-(2-methoxyphenyl)ureido]phenylacetate (2.44 g, quant.) as a
white crystalline material. mp 107-109.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.26 (t, J=7.1 Hz, 3H), 3.56 (s, 3H), 3.79 (s,
3H), 4.15 (q, J=7.1 Hz, 2H), 6.82-6.85 (n, 1H), 6.91-7.00 (m, 2H),
7.08 (s, 1H), 7.17 (d, J=8.5 Hz, 2H), 7.27 (d, J=8.6 Hz, 2H), 7.33
(s, 1H), 8.07-8.10 (m, 1H); MS (ESI) m/z 329 (M.sup.++H).
[0637] To a stirred solution of ethyl
4-[N'-(2-methoxyphenyl)ureido]phenylacetate (2.22 g, 6.78 mmol) in
MeOH (30 ml) was added 1.0 M-NaOH aq. (10.2 ml, 10.2 mmol), and the
resulting mixture was stirred overnight. 1N-HCl (aq.) was added and
the mixture was concentrated in vacuo. Water was added to the
residue to give precipitate, which was collected by filtration. The
crude solid was recrystallized from EtOH/hexane to give
4-[N'-(2-methoxyphenyl)ureido]phenylacetic acid as a white powder
(1.87 g, 92%). mp 165-168.degree. C.; .sup.1H-NMR (CD.sub.3OD)
.delta. 2.30 (s, 3H), 3.55 (s, 2H), 4.87 (s, 3H), 6.87-6.92 (m,
2H), 6.97-6.99 (m, 2H), 7.10-7.24 (m, 2H), 7.53-7.58 (m, 1H), 8.04
(d, J=7.2 Hz, 1H); MS (ESI) m/z 300 (M.sup.+).
[0638] To a stirred solution of
4-[N'-(2-methoxyphenyl)ureido]phenylacetic acid (353 mg, 1.18
mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (298
mg, 1.18 mmol) and N,N-dimethylaminopyridine (144 mg, 1.18 mmol) in
DMF (10 ml) was added EDC.HCl (226 mg, 1.18 mmol) at rt, and the
resulting mixture was stirred for 22 h. The mixture was poured into
1N-HCl aq. and extracted with EtOAc. The organic layer was washed
with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (10:1) to give methyl
4-[(4S)-fluoro1-[4-[N'-(2-methoxyphenyl)ureido]phenylacetyl]-(2S)--
pyrrolidinylmethoxy]benzoate (594 mg, 94%) as a colorless amorphous
solid. .sup.1H-NMR (CDCl.sub.3) mixture of rotamars, .delta.
2.05-2.58 (series of m, 2H), 3.55-4.25 (series of m, 5H), 3.77 (s,
3H), 3.87-3.90 (m, 3H), 4.50-4.63 (m, 2H), 5.23-5.37 (m, 1H),
6.81-6.84 (m, 1H), 6.91-6.99 (m, 4H), 7.09-7.12 (m, 2H), 7.18-7.26
(m, 2H), 7.45-7.53 (m, 2H), 7.91-8.03 (m, 2H), 8.10-8.12 (m, 1H);
MS (ESI) m/z 536 (M.sup.++H).
[0639] To a stirred solution of methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]phenylacetyl]-(2S)-pyrrol-
idinylmethoxy]benzoate (568 mg, 1.06 mmol) in MeOH-THF (1:1, 10 ml)
was added 1.0N-NaOH aq. (4.24 ml, 4.24 mmol) at rt, and the
resulting mixture was heated at 60.degree. C. for 1 h. The mixture
was poured into 1N-HCl aq. and extracted with EtOAc. The organic
layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give 85 (516 mg, 93%) as a colorless amorphous solid. MW 521.54
.sup.1H-NMR (CD.sub.3OD), mixture of rotamars, .delta. 2.12-2.46
(m, 2H), 3.65-4.19 (series of m, 5H), 3.88 (s, 3H), 4.42-4.45 (m,
1H), 4.52-4.62 (m, 1H), 5.24-5.39 (m, 1H), 6.85-6.91 (m, 1H),
6.94-7.03 (series of m, 4H), 7.14-7.19 (m, 2H), 7.35-7.40 (m, 2H),
7.92-7.96 (m, 2H), 8.02-8.04 (m, 1H); MS (ESI) m/z 521 (M.sup.++H),
544 (M.sup.++Na.sup.+).
Example 80
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]-3-methoxyphenylacetyl]-(2-
S)-pyrrolidinyl methoxy]benzoic acid
[0640] ##STR1707##
[0641] To a stirred solution of tertbutyl
4-amino-3-methoxy]phenylacetate (1.41 g, 5.94 mmol) and
triethylamine (165 ml, 1.19 mmol) in THF (20 ml) was added
2-methoxyphenyl isocyanate (790 ml, 5.94 mmol), and the resulting
mixture was stirred for 4 days. The mixture was concentrated to a
small volume in vacuo, and hexane was added to the residue to give
precipitate, which was collected to give tert-butyl
4-[N'-(2-methoxyphenyl)ureido]-3-methoxy]phenylacetate (2.06 g,
90%) as a white crystalline material. mp 132-134.degree. C.;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.46 (s, 9H), 3.50 (s, 2H), 3.87
(s, 3H), 3.88 (s, 3H), 6.84 (s, 1H), 6.87-6.90 (m, 2H), 6.98-7.03
(m, 2H), 7.12 (s, 1H), 7.16 (s, 1H), 8.06 (d, J=8.4 Hz, 1H), 8.13
(dd, J=7.2, 2.0 Hz, 1H); MS (ESI) m/z 387 (M.sup.++H).
[0642] To a stirred solution of tert-butyl
4-[N'-(2-methoxyphenyl)ureido]-3-methoxy]phenyl-acetate (2.01 g,
5.20 mmol) in CH.sub.2Cl.sub.2 (15 ml) was added trifluoroacetic
acid (10 ml), and the resulting mixture was heated under reflux for
30 min. The mixture was concentrated in vacuo. Water was added to
the residue to give precipitate, which was collected by filtration.
The crude solid was recrystallized from EtOH/hexane to give
4-[N'-(2-methoxyphenyl)ureido]-3-methoxy phenylacetic acid as white
powder (1.40 g, 82%). mp 182-185.degree. C.; .sup.1H-NMR
(CD.sub.3OD) .delta. 3.55 (s, 2H), 3.88 (s, 3H), 3.89 (s, 3H),
6.80-6.99 (m, 5H), 7.94 (d, J=8.4 Hz, 1H), 8.00 (d, J=7.2 Hz, 1H);
MS (ESI) m/z 330 (M.sup.+).
[0643] To a stirred solution of
4-[N'-(2-methoxyphenyl)ureido]-3-methoxyphenylacetic acid (353 mg,
1.07 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate
(271 mg, 1.07 mmol) and N,N-dimethylaminopyridine (131 mg, 1.07
mmol) in DMF (10 ml) was added EDC.HCl (224 mg, 1.18 mmol) at rt,
and the resulting mixture was stirred for 14 h. The mixture was
poured into 1N-HCl aq. and extracted with EtOAc. The organic layer
was washed with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (10:1) as eluent to give methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)
ureido]-3-methoxyphenylacetyl]-(2S)-pyrrolidinylmethoxy]benzoate
(372 mg, 61%) as a colorless amorphous solid. .sup.1H-NMR
(CDCl.sub.3), mixture of rotamars, .delta. 2.04-2.57 (series of m,
2H), 3.58-4.18 (series of m, 5H), 3.79 and 3.83 (s, total 3H), 3.86
(s, 3H), 3.87 (s, 3H), 4.51-4.63 (m, 2H), 5.22-5.36 (m, 1H),
6.80-6.89 (m, 3H), 6.94-7.03 (m, 4H), 7.15-7.25 (m, 2H), 7.94-8.01
(m, 2H), 8.04-8.11 (m, 2H); MS (ESI) m/z 566 (M.sup.++H).
[0644] To a stirred solution of methyl
4-[(4S)-fluoro-1-[4-[N'-(2-methoxyphenyl)ureido]-3-methoxy
phenyl-acetyl]-(2S)-pyrrolidinylmethoxy]benzoate (356 mg, 0.63
mmol) in MeOH-THF (1:1, 10 ml) was added 1.0N-NaOH aq. (1.88 ml,
1.88 mmol) at rt, and the resulting mixture was heated at
60.degree. C. for 2 h. The mixture was poured into 1N-HCl aq. and
extracted with EtOAc. The organic layer was washed with brine,
drying over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(10:1) as eluent to give 86 (335 mg, 97%) as a colorless amorphous
solid. MW 551.56 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars,
.delta. 2.14-2.48 (m, 2H), 3.69-4.20 (series of m, 5H), 3.88 (s,
3H), 3.89 (s, 3H), 4.46-4.57 (m, 2H), 5.27-5.41 (m, 1H), 6.79-7.04
(m, 7H), 7.90-8.02 (m, 4H); MS (ESI) m/z 552 (M.sup.++H).
Example 81
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-pyrr-
olidinyl]methoxy benzoic acid
[0645] ##STR1708##
[0646] To a mixture of ethyl 4-aminophenylacetate (1.62 g, 9.04
mmol) and 2,6-dichlorophenyl isocyanate (1.70 g, 9.04 mmol) in THF
(40 ml) was added Et.sub.3N (0.25 ml, 1.81 mmol) at room
temperature. After 2 h stirring, the reaction mixture was
concentrated in vacuo. The residue was triturated by the addition
of n-hexane, to give ethyl 4-[N'-(2,6-chlorophenyl)ureido]phenyl
acetate (3.19 g, 96%) as a colorless powder. mp 168-170.degree. C.
(dec.); .sup.1H-NMR (CDCl.sub.3) .delta. 1.25 (t, J=7.1 Hz, 3H),
3.56 (s, 2H), 4.14 (q, J=7.1 Hz, 2H), 6.50 (br, 1H), 6.67 (br, 1H),
7.12-7.52 (m, 7H).
[0647] To a stirred solution of ethyl
4-[N'-(2,6-dichlorophenyl)ureido]phenylacetate (3.19 g, 8.69 mmol)
in THF (70 ml), 0.25 N NaOH (70 ml) was added. After stirring at
room temperature for 17 h, the solvent was concentrated in vacuo.
The residue was triturated by the addition of water, to give
4-[N'-(2,6-dichlorophenyl)ureido]phenylacetic acid (2.44 g, 82%) as
colorless powder. mp 262-263.degree. C. (dec.); .sup.1H-NMR
(DMSO-d.sub.6) .delta. 3.48 (s, 2H), 7.14 (d, J=8.3 Hz, 2H), 7.31
(t, J=8.3 Hz, 1H), 7.37 (d, J=8.3 Hz, 2H), 7.52 (d, J=8.0 Hz, 2H),
8.18 (s, 1H), 8.90 (s, 1H), 12.22 (br, 1H); MS (ESI) m/z 339
(M.sup.+30 1), 341 (M.sup.++3), 343 (M.sup.++5).
[0648] A mixture of 4-[N'-(2,6-dichlorophenyl)ureido]phenylacetic
acid (268 mg, 0.79 mmol), methyl
4-[(2S,4S)-4-fluoro-2-pyrrolidinyl]methoxybenzoate (200 mg, 0.79
mmol), EDC.HCl (227 mg, 1.19 mmol), HOBT (161 mg, 1.19 mmol) and
Et.sub.3N (0.55 ml, 3.95 mmol) in DMF (4 ml) was stirred at room
temperature for 18 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was purified on TLC
[CHCl.sub.3/MeOH (10/1)], to give trimethyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-pyr-
rolidinyl]methoxybenzoate (465 mg, 100%) as a colorless amorphous
solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.05-2.57 (m, 2H), 3.60 (d,
2H, J=3.4 Hz), 3.64-3.84 (m, 2H), 3.88 and 3.89 (each s, 3H, amide
isomers), 3.92-4.63 (m, 3H), 5.22-5.38 (m, 1H), 6.87 and 6.89 (each
d, each J=7.9 Hz, 2H, amide isomers), 7.01-7.17 (m, 6H), 7.28 (m,
2H), 7.36 (br, 1H), 7.92 (d, J=8.8 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H);
MS (ESI) m/z 574 (M.sup.++1), 576 (M.sup.++3), 578 (M.sup.++5).
[0649] To a solution of methyl
4-[(2S,4S)-1-[4-[N'-(2,6-dichlorophenyl)ureido]phenylacetyl]-4-fluoro-2-p-
yrrolidinyl]methoxybenzoate (465 mg, 0.809 mmol) in THF (40 ml),
0.25 N NaOH (40 ml) was added. After stirring at room temperature
for 11 h, the mixture was acidified with 1 N HCl and extracted with
CHCl.sub.3-MeOH (10/1). The combined extracts were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified on TLC [CHCl.sub.3/MeOH (10/1)], to give 87 (340 mg, 75%)
as a colorless powder. MW 560.40 mp 168-172.degree. C. (dec.); IR
(KBr) 3340, 1711, 1685, 1604, 1240, 773 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.22-2.30 (m, 2H), 3.61 (d, J=7.4 Hz, 2H),
3.70-4.75 (m, 6H), 5.30-5.49 (m, 1H), 7.02-7.18 (m, 5H), 7.28-7.41
(m, 4H), 7.52 (dd, J=8.0, 2.9 Hz, 2H), 7.86 (m, 2H), 8.29 (br, 1H),
9.01 (br, 1H), 12.66 (br, 1H); MS (ESI) m/z 560 (M.sup.++1), 562
(M.sup.++3), 564 (M.sup.++5); Anal. Calcd for
C.sub.27H.sub.24Cl.sub.2FN.sub.3O.sub.5.0.5H.sub.2O: C, 56.95; H,
4.43; Cl, 12.45; F, 3.34; N, 7.38. Found: C, 57.04; H, 4.34; Cl,
12.98; F, 3.27; N, 7.21.
Example 82
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluoro-
-(2S)-pyrrolidinyl]methoxybenzoic acid
[0650] ##STR1709##
[0651] To a mixture of tertbutyl 4-amino-3-methoxyphenylacetate
(2.15 g, 9.04 mmol), 2,6-dichlorophenyl isocyanate (1.70 g, 9.04
mmol) in THF (40 ml) was added Et.sub.3N (0.25 ml, 9.04 mmol) at
room temperature. After 18 h stirring, the reaction mixture was
concentrated in vacuo. The residue was triturated by the addition
of n-hexane, to give tert-butyl
4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetate (2.27 g,
59%) as a colorless powder. mp 177-181.degree. C. (dec.);
.sup.1H-NMR (CDCl.sub.3) .delta. 1.43 (s, 9H), 3.74 (s, 2H), 3.83
(s, 3H), 6.34 (s, 1H), 6.81 (s, 1H), 6.84 (d, J=8.3 Hz, 1H), 7.06
(r, 1H), 7.27 (t, J=8.1 Hz, 1H), 7.39 (d, J=8.1 Hz, 2H), 8.04 (d,
J=8.3 Hz, 1H).
[0652] To a stirred solution of tertbutyl
4-[N'-(2,6-chlorophenyl)ureido]-3-methoxyphenylacetate (2.27 g,
5.34 mmol) in CH.sub.2Cl.sub.2 (50 ml) was added TFA (20 ml) at
room temperature. After 2 h stirring, the mixture was concentrated
in vacuo. The residue was triturated by the addition of water, to
give 4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetic acid
(1.50 g, 76%) as a colorless powder. mp 246-249.degree. C. (dec.);
.sup.1H-NMR (DMSO-d.sub.6) .delta. 3.49 (s, 2H), 3.88 (s, 3H), 6.75
(d, J=8.3 Hz, 1H), 6.93 (s, 1H), 7.30 (t, J=7.8 Hz, 1H), 7.52 (d,
J=8.0 Hz, 2H), 7.97 (d, J=8.0 Hz, 1H), 8.40 (s, 1H), 8.86 (s, 1H),
12.23 (br, 1H); MS (ESI) m/z 369 (M.sup.++1), 371 (M.sup.++3), 373
(M.sup.++5).
[0653] A mixture of
4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetic acid (288
mg, 0.78 mmol), methyl
4-[(2S,4S)-4-fluoro-2-pyrrolidinyl]methoxybenzoate (200 mg, 0.79
mmol), EDC.HCl (227 mg, 1.19 mmol), HOBT (161 mg, 1.19 mmol) and
Et.sub.3N (0.55 ml, 3.95 mmol) in DMF (4 ml) was stirred at room
temperature for 18 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/MeOH (40/1)] to give methyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenyl
acetyl]-(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate (530 mg,
100%) as a colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 2.03-2.62 (m, 2H), 3.61 (d, 2H, J=4.7 Hz), 3.62-3.66 (m,
2H), 3.73 and 3.77 (each s, 3H, amide isomers), 3.78-3.85 (m, 1H),
3.87 and 3.88 (each s, 3H, amide isomers), 3.95-4.63 (m, 4H),
5.22-5.36 (m, 1H), 6.82 (s, 1H), 6.88 (d, J=8.8 Hz, 1H), 6.95 (d,
J=8.8 Hz, 2H), 7.14-7.25 (m, 1H), 7.38 (d, J=8.1 Hz, 2H), 7.94-8.10
(m, 3H); MS (ESI) m/z 604 (M.sup.++1), 606 (M.sup.++3), 608
(M.sup.++5).
[0654] To a solution of methyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-fluor-
o-(2S)-pyrrolidinyl]methoxybenzoate (530 mg, 0.78 mmol) in THF (40
ml), 0.25 N NaOH (40 ml) was added. After stirring at room
temperature for 11 h, the mixture was acidified with 1 N HCl and
extracted with CHCl.sub.3-MeOH (10/1). The combined extracts were
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was purified on TLC [CHCl.sub.3/MeOH (10/1)] to give 88 (420 mg,
75%) as a colorless amorphous solid. MW 590.43 mp 162-168.degree.
C. (dec.); IR (KBr) 3346, 2974, 1709, 1604, 1533, 1254 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.98-2.36 (m, 2H), 3.58 (s, 2H),
3.78-3.95 (m, 6H), 4.02-4.68 (m, 2H), 5.31-5.50 (m, 1H), 6.71-7.09
(m, 4H), 7.31 (t, J=7.8 Hz, 1H), 7.53 (d, J=8.1 Hz, 2H), 7.87 (d,
J=8.1 Hz, 2H), 7.88-8.00 (m, 1H), 8.30-8.40 (m, 1H), 8.89 (s, 1H);
MS (ESI) m/z 590 (M.sup.++1), 592 (M.sup.++3), 594 (M.sup.++5);
Anal. Calcd for
C.sub.28H.sub.26Cl.sub.2FN.sub.3O.sub.6.1.5H.sub.2O: C, 54.47; H,
4.73; F, 3.08; N, 6.81. Found: C, 54.53; H, 4.49; F, 2.93; N,
6.65.
Example 83
4-[(2S,4S)-1-[4-[N'-(2,6-Dichlorophenyl)ureido]-3-methylphenylacetyl]-4-fl-
uoro-2-pyrrolidinyl]methoxybenzoic acid
[0655] ##STR1710##
[0656] To a mixture of tert-butyl 4-amino-3-methylphenylacetate
(1.88 g, 8.51 mmol), 2,6-dichlorophenyl isocyanate (1.60 g, 8.51
mmol) in THF (40 ml) was added Et.sub.3N (0.24 ml, 1.70 mmol) at
room temperature. After 3 h stirring, the reaction mixture was
concentrated in vacuo. The residue was triturated by the addition
of n-hexane, to give tert-butyl
4-[N'-(2,6-dichlorophenyl)ureido]-3-methylphenylacetate (2.58 g,
74%) as a colorless powder. mp 243-244.degree. C. (dec.);
.sup.1H-NMR (CDCl.sub.3) .delta. 1.45 (s, 9H), 2.30 (s, 3H), 3.49
(s, 2H), 6.24 (s, 2H), 7.12-7.16 (m, 3H), 7.35 (d, J=8.3 Hz, 2H),
7.51 (d, J=7.8 Hz, 1H).
[0657] To a stirred solution of tert-butyl
4-[N'-(2,6-dichlorophenyl)ureido]-3-methylphenylacetate (2.58 g,
6.30 mmol) in CH.sub.2Cl.sub.2 (50 ml) was added TFA (20 ml) at
room temperature. After 2 h stirring, the mixture was concentrated
in vacuo. The residue was triturated by the addition of water, to
give 4-[N'-(2,6-dichlorophenyl)ureido]-3-methylphenylacetic acid
(2.12 g, 95%) as a colorless powder. mp 274-283.degree. C. (dec.);
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24 (s, 3H), 3.46 (s, 2H), 7.00
(d, J=8.6 Hz, 1H), 7.06 (s, 1H), 7.30 (t, J=7.8 Hz, 1H), 7.52 (d,
J=8.3 Hz, 2H), 7.65 (d, J=8.2 Hz, 1H), 8.12 (s, 1H), 8.50 (s, 1H),
12.22 (br, 1H); MS (ESI) m/z 353 (M.sup.++1), 355 (M.sup.++3), 357
(M.sup.++5).
[0658] A mixture of
4-[N'-(2,6-dichlorophenyl)ureido]-3-methylphenylacetic acid (181
mg, 0.51 mmol), methyl
4-[(2S,4S)-4-fluoro-2-pyrrolidinyl]methoxybenzoate (130 mg, 0.51
mmol), EDC.HCl (147 mg, 0.77 mmol), HOBT (104 mg, 0.77 mmol) and
Et.sub.3N (0.35 ml, 2.55 mmol) in DMF (4 ml) was stirred at room
temperature for 18 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was purified on TLC
[CHCl.sub.3/MeOH (20 .mu.l)], to give methyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methylphenylacetyl]-(4S)-fluoro-
-(2S)-pyrrolidinyl]methoxybenzoate (283 mg, 94%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.95-2.61 (m,
3H), 3.55 (br, 2H), 3.67-3.81 (m, 2H), 3.87 (s, 6H), 3.89-4.68 (m,
2H), 5.23-5.43 (m, 1H), 6.81-7.10 (m, 6H), 7.13-7.43 (m, 3H), 7.56
(br, 1H, one of isomers), 7.73 (br, 1H, one of isomers), 7.89-8.00
(m, 2H); MS (ESI) m/z 588 (M.sup.++1), 590 (M.sup.++3), 592
(M.sup.++5).
[0659] To a solution of methyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methylphenylacetyl]-(4S)-fluoro-
-(2S)-pyrrolidinyl]methoxybenzoate (283 mg, 0.48 mmol) in THF (20
ml), 0.25 N NaOH (20 ml) was added. After stirring at room
temperature for 11 h, the mixture was extracted with EtOAc. The
remaining aqueous layer was acidified with 1 N HCl and extracted
with EtOAc. The combined extracts were dried over Na.sub.2SO.sub.4
and concentrated in vacuo. The residue was on TLC [CHCl.sub.3/MeOH
(20/1)] to give 89 (450 mg, 67%) as a pale brown amorphous solid.
MW 574.43 mp 174-180.degree. C. (dec.); IR (KBr) 3330, 3288, 1711,
1685, 1604, 1512, 1242 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.24 (m, 3H), 3.61 (d, 2H, J=6.1 Hz), 3.72-4.68 (m, 7H),
5.30-5.50 (m, 1H), 6.97-7.20 (m, 4H), 7.29-7.68 (m, 5H), 7.87 (m,
2H), 8.10-8.95 (m, 1H), 12.65 (br, 1H); MS (ESI) m/z 574
(M.sup.++1), 576 (M.sup.++3), 578 (M.sup.++5); Anal. Calcd for
C.sub.28H.sub.26Cl.sub.2FNO.sub.3O.sub.5.0.5H.sub.2O: C, 57.64; H,
4.66; Cl, 12.15; F, 3.26; N, 7.20. Found: C, 57.37; H, 4.44; Cl,
12.64; F, 3.23; N, 7.25.
Example 84
4-[1-[3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoic acid
[0660] ##STR1711##
[0661] To a stirred solution of 3-chlorophenylacetic acid (21.76 g,
127.6 mmol) in dichloroethane (100 ml) was added MeOH (15.6 ml, 383
mmol) and H.sub.2SO.sub.4 (1 ml) at room temperature. After 20
minutes stirring, the mixture was heated at 80.degree. C. for 2 h.
The reaction mixture was poured into ice water and extracted with
CHCl.sub.3. The combined extracts were washed with aq. NaHCO.sub.3
and brine. After dried over Na.sub.2SO.sub.4, the extract was
concentrated in vacuo to give methyl 3-chlorophenylacetate (25.4 g,
100%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.60 (s,
2H), 3.70 (s, 3H), 7.15-7.26 (m, 4H).
[0662] To a stirred mixture of methyl 3-chlorophenylacetate (25.4
g, 128 mmol) in H.sub.2SO.sub.4 (44 ml) was added HNO.sub.3 (5.5
ml, 138 mmol) at 0.degree. C. The reaction mixture was gradually
raised to room temperature for 4 h. The reaction mixture was poured
into ice water and extracted with EtOAc. The combined extracts were
washed with aq. NaHCO.sub.3 and brine. After dried over
Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [1 kg, n-hexane/EtOAc
(40/1)] to give methyl 3-chloro-4-nitrophenylacetate (11.4 g, 36%)
as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.69 (s, 2H),
3.74 (s, 3H), 7.33 (dd, J=8.3, 1.5 Hz, 1H), 7.49 (d, J=1.5 Hz, 1H),
7.87 (d, J=8.3 Hz, 1H).
[0663] A mixture of methyl 3-chloro-4-nitrophenylacetate (10.9 g,
47.5 mmol), reduced iron powder (8.58 g, 153.6 mmol),
AcONa.3H.sub.2O (6.05 g, 44.5 mmol) and AcOH (17.6 ml) in
MeOH/H.sub.2O (100/400 ml) was heated at 110.degree. C. for 1 h.
After cooled to room temperature, the reaction mixture was filtered
through Celite and the filtered cake was washed with MeOH, The
combined filtrate were evaporated and extracted with EtOAc. The
combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
chromatographed on silica gel [150 g, CHCl.sub.3/EtOAc (10/1)] to
give methyl 4-amino-3-chlorophenylacetate (4.58 g, 48%) as a red
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.49 (s, 2H), 3.68 (s, 3H),
4.01 (br, 2H), 6.70 (d, J=7.4 Hz, 1H), 6.96 (dd, J=8.1, 2.0 Hz,
1H), 7.17 (d, J=2.0 Hz, 1H).
[0664] To a mixture of methyl 4-amino-3-chlorophenylacetate (1.00
g, 5.01 mmol) and 2-methylphenyl isocyanate (0.60 ml, 5.01 mmol) in
THF (20 ml) was added Et.sub.3N (0.14 ml, 1.00 mmol) at room
temperature. After 1 day stirring, 2-methylphenyl isocyanate (0.60
ml, 5.01 mmol) was added to the reaction mixture and stirred 17 h.
The reaction mixture was concentrated in vacuo. The residue was
triturated by the addition of n-hexane to give methyl
3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetate (1.23 g, 74%)
as a colorless powder. .sup.1H-NMR (CDCl.sub.3) .delta. 2.34 (s,
3H), 3.54 (s, 2H), 3.68 (s, 3H), 6.24 (br, 1H), 6.99 (br, 1H), 7.15
(dd, J=8.3, 2.0 Hz, 1H), 7.21-7.31 (m, 5H), 7.44 (d, J=7.6 Hz, 1H),
8.20 (d, J=8.5 Hz, 1H); MS (ESI) m/z 333 (M.sup.++1), 335
(M.sup.++3).
[0665] To a stirred solution of methyl
3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetate (1.23 g, 3.70
mmol) in THF (30 ml) was added 0.25 N NaOH (30 ml). After stirring
at room temperature for 14 h, the solvent was concentrated in
vacuo. The residue was triturated by the addition of 1 N HCl and
dried over 60.degree. C. for 2 days under a reduced pressure to
give 3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.22
g, 100%) as colorless powder. .sup.1H-NMR (DMSO-d.sub.6) 6.26 (s,
3H), 3.40 (s, 2H), 6.95 (t, J=7.3 Hz, 1H), 7.11 (d, J=7.6 Hz, 2H),
7.16 (d, J=7.3 Hz, 1H), 7.32 (s, 1H), 7.76 (d, J=8.0 Hz, 1H), 7.94
(dd, J=9.3, 1.0 Hz, 1H), 8.72 (s, 2H); MS (ESI) m/z 319
(M.sup.++1), 321 (M.sup.++3), 341 (M.sup.++Na).
[0666] A mixture of
3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (319 mg,
1.00 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate
(253 mg, 1.00 mmol), EDC.HCl (288 mg, 1.50 mmol), HOBT (203 mg,
1.50 mmol) and Et.sub.3N (0.70 ml, 5.00 mmol) in DMF (4 ml) was
stirred at room temperature for 15 h. The mixture was poured into
ice water and extracted with EtOAc. The combined extracts were
washed with ice water and brine. After dried over Na.sub.2SO.sub.4,
the extracts were concentrated in vacuo. The residue was purified
on TLC [CHCl.sub.3/acetone (5/1)] to give methyl
4-[1-[3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinyl]methoxybenzoate (480 mg, 87%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.10-2.60 (m,
2H), 2.29 (s, 3H), 3.56 (d, J=6.8 Hz, 1H), 3.71-3.84 (m, 1H), 3.87
and 3.89 (each s, 3H, amide isomers), 3.91-4.20 (m, 3H), 4.49-4.60
(m, 2H), 5.32 (dt, J=53.0, 4.2 Hz, 1H), 6.80 (br, 1H), 6.89 and
6.95 (each d, J=8.8 Hz, 2H, amide isomers), 7.09-7.26 (m, 6H), 7.50
(d, J=7.3 Hz, 1H), 7.94 and 8.00 (each d, J=8.8 Hz, 2H, amide
isomers), 8.10 and 8.15 (each d, J=8.3 Hz, 1H, amide isomers); MS
(FAB) m/z 554 (M.sup.++1), 556 (M.sup.++3).
[0667] To a solution of methyl
4-[1-[3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinyl]methoxybenzoate (480 mg, 0.866 mmol) in THF (30 ml)
was added 0.25 N NaOH (30 ml). After stirring at room temperature
for 2 days, the mixture was concentrated under a reduced pressure
and acidified with 1 N HCl. The precipitates were collected, washed
with water and dried under a reduced pressure to give 90 (374 mg,
80%) as a colorless powder. MW 539.98 IR (KBr) 3354, 3060, 2976,
1709, 1604, 1244 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.27
(s, 3H), 2.31 (s, 2H), 3.66 (d, J=7.2 Hz, 2H), 3.71-4.67 (m, 5H),
5.32-5.53 (m, 1H), 6.97 (t, J=7.3 Hz, 1H), 7.04-7.22 (m, 5H), 7.32
and 7.35 (each d, J=1.7 Hz, 1H, amide isomers), 7.77 (d, J=7.6 Hz,
1H), 7.87 and 7.90 (each d, J=9.0 Hz, 2H, amide isomers), 8.01 and
8.03 (each d, J=8.5 Hz, 1H, amide isomers), 8.57 and 8.59 (each s,
1H, amide isomers), 8.63 and 8.65 (each s, 1H, amide isomers),
12.63 (s, 1H); MS (ESI) m/z 540 (M.sup.++1), 542 (M.sup.++3).
Example 85
4-[1-[3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxy benzoic acid
[0668] ##STR1712##
[0669] To a mixture of methyl 4-amino-3-chlorophenylacetate (1.00
g, 5.01 mmol) and 2-chlorophenyl isocyanate (0.60 ml, 5.01 mmol) in
THF (20 ml) was added Et.sub.3N (0.14 ml, 1.00 mmol) at room
temperature. After 1 day stirring, 2-chlorophenyl isocyanate (0.60
ml, 5.01 mmol) was added to the reaction mixture and stirred 17 h.
The reaction mixture was concentrated in vacuo. The residue was
triturated by the addition of n-hexane to give methyl
3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetate (1.35 g, 76%)
as a colorless powder. .sup.1H-NMR (CDCl.sub.3) .delta. 3.58 (s,
3H), 3.71 (s, 2H), 7.04 (m, 3H), 7.18 (dd, J=8.5, 2.0 Hz, 1H),
7.27-7.39 (m, 3H), 8.07 (m, 2H); MS (ESI) m/z 353 (M.sup.++1), 355
(M.sup.++3), 357 (M.sup.++5).
[0670] To a stirred solution of methyl
3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetate (1.35 g, 3.82
mmol) in THF (30 ml) was added 0.25 N NaOH (30 ml). After stirring
at room temperature for 14 h, the solvent was concentrated in
vacuo. The residue was triturated by the addition of 1 N HCl and
dried at 60.degree. C. for 2 days under a reduced pressure to give
3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetic acid (1.12 g,
86%) as colorless powder. .sup.1H-NMR (DMSO-d.sub.6) .delta. 3.52
(s, 2H), 7.05 (m, 1H), 7.17 (d, J=8.5 Hz, 1H), 7.30 (d, J=7.6 Hz,
1H), 7.37 (s, 1H), 7.46 (dd, J=8.0, 1.5 Hz, 1H), 7.95 (dd, J=8.3,
1.2 Hz, 1H), 8.07 (d, J=8.3 Hz, 1H), 9.00 (d, J=8.0 Hz, 2H); MS
(FAB) m/z 339 (M.sup.++1), 341 (M.sup.++3), 343 (M.sup.++5).
[0671] A mixture of
3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetic acid (339 mg,
1.00 mmol), methyl
4-[(2S,4S)-4-fluoro-2-pyrrolidinyl]methoxybenzoate (253 mg, 1.00
mmol), EDC.HCl (288 mg, 1.50 mmol), HOBT (203 mg, 1.50 mmol) and
Et.sub.3N (0.70 ml, 5.00 mmol) in DMF (4 ml) was stirred at room
temperature for 15 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/acetone (10/1)] to give methyl
4-[1-[3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinyl]methoxybenzoate (550 mg, 96%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.14-2.64 (m,
2H), 3.59 (d, J=11.2 Hz, 2H), 3.78-3.82 (m, 1H), 3.86 and 3.89
(each s, 3H, amide isomers), 3.91-4.28 (m, 2H), 4.50-4.79 (m, 2H),
5.34 and 5.39 (each dt, J=52.5, 4.4 Hz, 1H, amide isomers),
6.89-6.98 (m, 3H), 7.09-7.13 (m, 2H), 7.22 (dt J=7.3, 2.2 Hz, 1H),
7.29 (dd, J=8.1, 2.0 Hz, 1H), 7.79 and 7.86 (each s, 1H, amide
isomers), 7.86-8.03 (m, 4H), 8.11 (dd, J=8.3, 1.0 Hz, 1H); MS (FAB)
m/z 574 (M.sup.+30 1), 576 (M.sup.++3), 578 (M.sup.++5).
[0672] To a solution of methyl
4-[1-[3-chloro-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S-
)-pyrrolidinyl]methoxybenzoate (550 mg, 0.957 mmol) in THF (30 ml)
was added 0.25 N NaOH (30 ml). After stirring at room temperature
for 2 days, the mixture was concentrated under a reduced pressure
and acidified with 1 N HCl. The precipitates were collected, washed
with water and dried under a reduced pressure to give 91 (437 mg,
82%) as a colorless powder. MW 560.40 IR (KBr) 3348, 3072, 2954,
1703, 1604, 1529, 1439 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.25-2.42 (m, 2H), 3.67 (d, J=8.3 Hz, 2H), 3.81-4.68 (m,
5H), 5.39 and 5.46 (each d, J=54.4 Hz, 1H, amide isomers),
7.04-7.10 (m, 3H), 7.18 (d, J=8.3 Hz, 1H), 7.31 (t, J=8.3 Hz, 1H),
7.33 and 7.37 (each s, 1H, amide isomers), 7.47 (d, J=8.1 Hz, 1H),
7.88 (dd, J=9.0, 3.2 Hz, 2H), 7.98 (dd, J=8.5, 3.0 Hz, 1H) m, 1H),
8.09 (d, J=8.3 Hz, 1H), 8.99 (d, J=2.9 Hz, 1H), 9.02 (s, 1H), 12.64
(s, 1H); MS (ESI) m/z 560 (M.sup.++1), 562 (M.sup.++3), 564
(M.sup.++5); Anal. Calcd for
C.sub.27H.sub.24Cl.sub.2FN.sub.3O.sub.5.0.2H.sub.2O: C, 57.50; H,
4.36; N, 7.45; Cl, 12.57; F, 3.37. Found: C, 57.72; H, 4.47; N,
7.14; Cl, 12.44; F, 3.44.
Example 86
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinyl]methoxybenzoic acid
[0673] ##STR1713##
[0674] To a mixture of methyl 4-amino-3-chlorophenylacetate (1.00
g, 5.01 mmol) and 2-bromophenyl isocyanate (0.62 ml, 5.01 mmol) in
THF (20 ml) was added Et.sub.3N (0.14 ml, 1.00 mmol) at room
temperature. After 1 day stirring, 2-bromophenyl isocyanate (0.60
ml, 5.01 mmol) was added to the reaction mixture and stirred 24 h.
The reaction mixture was concentrated in vacuo. The residue was
triturated by the addition of n-hexane to give methyl
4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetate (1.34 g, 67%) as
a colorless powder. .sup.1H-NMR (CDCl.sub.3) .delta. 3.58 (s, 3H),
3.70 (s, 2H), 6.98 (m, 3H), 7.19 (dd, J=8.3, 1.9 Hz, 1H), 7.32 (m,
1H), 7.51 (m, 2H), 8.05 (m, 1H); MS (ESI) m/z 398 (M.sup.++1), 400
(M.sup.++3), 402 (M.sup.++5).
[0675] To a stirred solution of methyl
4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetate (1.34 g, 3.37
mmol) in THF (30 ml) was added 0.25 N NaOH (30 ml). After stirring
at room temperature for 14 h, the solvent was concentrated in
vacuo. The residue was triturated by the addition of 1 N HCl and
dried at 60.degree. C. for 2 days under a reduced pressure to give
4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetic acid (1.03 g,
80%) as colorless powder. .sup.1H-NMR (DMSO-d.sub.6) .delta. 3.56
(s, 2H), 7.00 (m, 1H), 7.17 (dd, J=9.0, 1.7 Hz, 1H), 7.32-7.40 (m,
2H), 7.62 (dd, J=8.0, 1.2 Hz, 1H), 7.95 (m, 2H), 8.83 (s, 1), 9.01
(s, H), 12.41 (br, 1H); MS (FAB) m/z 385 (M.sup.++2), 386
(M.sup.++4), 388 (M.sup.++6).
[0676] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetic acid (384 mg,
1.00 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate
(253 mg, 1.00 mmol), EDC.HCl (288 mg, 1.50 mmol), HOBT (203 mg,
1.50 mmol) and Et.sub.3N (0.70 ml, 5.00 mmol) in DMF (4 ml) was
stirred at room temperature for 15 h. The mixture was poured into
ice water and extracted with EtOAc. The combined extracts were
washed with ice water and brine. After dried over Na.sub.2SO.sub.4,
the extracts were concentrated in vacuo. The residue was
chromatographed on silica gel [50 g, CHCl.sub.3/acetone (10/1)] to
give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoate (530 mg, 86%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta.2.14-2.63 (m, 2H),
3.58 (d, J=10.0 Hz, 1H), 3.73-3.83 (m, 1H), 3.86 and 3.89 (each s,
3H, amide isomers), 3.90-4.29 (m, 3H), 4.50-4.69 (m, 2H), 5.33 and
5.37 (each m, 1H, amide isomers), 6.88-6.93 (m, 3H), 7.11-7.14 (m
2H), 7.26 (m, 1H), 7.46 (d, J=8.1 Hz, 1H), 7.62-7.78 (m, 2H), 7.89
and 7.93 (each m, 2H, amide isomers), 8.01 (dd, J=8.8, 1.7 Hz, 2H);
MS (FAB) m/z 618 (M.sup.+), 620 (M.sup.++2), 622 (M.sup.++4).
[0677] To a solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-chlorophenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoate (530 mg, 0.856 mmol) in THF (30 ml)
was added 0.25 N NaOH (30 ml). After stirring at room temperature
for 2 days, the mixture was concentrated under a reduced pressure
and acidified with 1 N HCl. The mixture was extracted with
CHCl.sub.3/MeOH (10/1). The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [20 g, CHCl.sub.3/acetone (10/1)-CHCl.sub.3/MeOH (10/1)]
to give 92 (59 mg, 11%) as a colorless amorphous solid. MW 604.85
IR (KBr) 3329, 3060, 2976, 1712, 1526, 1435 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.31 (m, H), 3.48-4.68 (m, 7H), 5.32-5.53
(m, 1H), 6.99-7.19 (m, 4H), 7.36 (s, 1H), 7.63 (dd, J=6.7, 1.2 Hz,
1H), 7.86-8.18 (m, 4H), 8.83 (s, 1H), 9.02 (s, 1H), 12.67 (br, 1H);
MS (ESI) m/z 604 (M.sup.++1), 606 (M.sup.++3), 608 (M.sup.++5);
Anal. Calcd for C.sub.27H.sub.24BrClFN.sub.3O.sub.5.0.5H.sub.2O: C,
52.83; H, 4.10; N, 6.85; Cl, 5.78; F, 3.09. Found: C, 53.24; H,
4.32; N, 6.43; Cl, 6.01; F, 3.07.
Example 87
4-[1-[3-chloro-4-(N'-phenylureido)phenylacetyl]-(4S)-fluoro-(2S)-pyrrolidi-
nyl]methoxybenzoic acid
[0678] ##STR1714##
[0679] To a mixture of methyl 4-amino-3-chlorophenylacetate (1.31
g, 6.56 mmol) and phenyl isocyanate (0.71 ml, 6.56 mmol) in THF (20
ml) was added Et.sub.3N (0.19 ml, 1.33 mmol) at room temperature.
After 15 h stirring, the reaction mixture was concentrated in
vacuo. The residue was triturated by the addition of n-hexane to
give methyl 3-chloro-4-(N'-phenylureido)phenylacetate (1.79 g, 86%)
as a pale brown solid. .sup.1H-NMR (CDCl.sub.3) .delta. 3.56 (s,
2H), 3.70 (s, 3H), 6.70 (m, 1H), 7.06 (s, 1H), 7.14-7.18 (m, 2H),
7.26 (dd, J=7.8, 1.9 Hz, 1H), 7.33-7.38 (m, 4H), 8.14 (dd, J=8.3,
3.0 Hz, 1H); MS (ESI) m/z 319 (M.sup.++1), 321 (M.sup.++3).
[0680] To a stirred solution of methyl
3-chloro-4-(N'-phenylureido)phenylacetate (1.79 g, 5.62 mmol) in
THF (30 ml) was added 0.25 N NaOH (30 ml). After stirring at room
temperature for 20 k, the solvent was concentrated in vacuo. The
residue was triturated by the addition of 1 N HCl and dried at
60.degree. C. for 2 days under a reduced pressure to give
3-chloro-4-(N'-phenylureido)phenylacetic acid (1.58 g, 92%) as pale
brown solid. .sup.1H-NMR (DMSO-d.sub.6) .delta. 3.55 (s, 2H), 6.99
(t, J=7.3 Hz, 1H), 7.17 (d, J=8.3 Hz, 1H), 7.29 (t, J=7.6 Hz, 2H),
7.36 (s, 1H), 7.46 (d, J=8.0 Hz, 2H), 8.07 (d, J=8.3 Hz, 1H), 8.28
(s, 1H), 9.37 (s, 1H), 12.37 (br, 1H).
[0681] A mixture of 3-chloro-4-(N'-phenylureido)phenylacetic acid
(305 mg, 1.00 mmol), methyl
4-[(2S,4S)-4-fluoro-2-pyrrolidinyl]methoxybenzoate (253 mg, 1.00
mmol), EDC.HCl (288 mg, 1.50 mmol), HOBT (203 mg, 1.50 mmol) and
Et.sub.3N (0.70 ml, 5.00 mmol) in DMF (4 ml) was stirred at room
temperature for 17 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [30 g, CHCl.sub.3/acetone (20/1)] to give methyl
4-[1-[3-chloro-4N'-phenylureido)phenylacetyl]-(4S)-fluoro-(2S)-pyrrolidin-
yl]methoxybenzoate (720 mg, 100%) as a colorless amorphous solid.
MS (FAB) m/z 540 (M.sup.++1), 542 (M.sup.++3).
[0682] To a solution of methyl
4-[1-[3-chloro-4-(N'-phenylureido)phenylacetyl]-(4S)-fluoro-(2S)-pyrrolid-
inyl]methoxybenzoate (720 mg, 1.00 mmol) in THF/MeOH (30/30 ml) was
added 0.25 N NaOH (30 ml). After stirring at room temperature for 2
h, the reaction mixture was heated at 50.degree. C. for 22 h. After
removed the solvent, the resulting residue was acidified with 1 N
HCl. The precipitates were collected, washed with water and dried
under a reduced pressure to give 93 [412 mg, 78% (2 Steps)] as a
colorless powder. MW 525.96 IR (KBr) 3346, 3302, 2976, 1712, 1604,
1240 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25-2.31 (r,
2H), 3.66 (d, J=7.8 Hz, 2H), 3.71-4.67 (m, 5H), 5.31-5.52 (m, 1H),
6.99 (t, J=7.3 Hz, 1H), 7.04 and 7.07 (each d, J=8.7 Hz, 2H, amide
isomers), 7.14-7.18 (m, 1H), 7.29 (t, J=7.3 Hz, 2H), 7.35 (d, J=1.7
Hz, 1H), 7.46 (d, J=7.8 Hz, 2H), 7.87 and 7.90 (each d, J=9.0 Hz,
2H, amide isomers), 8.04 and 8.06 (each d, J=8.5 Hz, 1H, amide
isomers), 8.26 and 8.28 (each s, 1H, amide isomers), 9.36 (s, 1H),
12.63 (s, 1H); MS (ESI) m/z 526 (M.sup.++1), 528 (M.sup.++3); Anal.
Calcd for C.sub.27H.sub.25ClFN.sub.3O.sub.5.0.5H.sub.2O: C, 60.62;
H, 4.90; N, 7.85; Cl, 6.63; F, 3.55. Found: C, 61.00; H, 5.19; N,
7.40; Cl, 6.66; F, 3.39.
Example 88
4-[1-[3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinyl]methoxybenzoic acid
[0683] ##STR1715##
[0684] To a stirred solution of 3-bromophenylacetic acid (10.2 g,
47.4 mmol) in dichloroethane (50 ml) was added MeOH (5.8 ml, 142
mmol) and H.sub.2SO.sub.4 (0.5 ml) at room temperature. After 20
minutes stirring, the mixture was heated at 80.degree. C. for 7 h.
The reaction mixture was poured into ice water and extracted with
CHCl.sub.3. The combined extracts were washed with aq. NaHCO.sub.3
and brine. After dried over Na.sub.2SO.sub.4, the extracts were
concentrated in vacuo to give methyl 3-bromophenyl acetate (10.8 g,
99%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.60 (s,
2H), 3.71 (d, J=1.0 Hz, 3H), 7.18-7.44 (m, 4H).
[0685] To a stirred mixture of methyl 3-chlorophenylacetate (10.8
g, 47.1 mmol) in H.sub.2SO.sub.4 (15.1 ml) was added HNO.sub.3 (2.8
ml, 70.7 mmol) at 0.degree. C. The reaction mixture was gradually
raised to room temperature for 5.5 h. The reaction mixture was
poured into ice water and extracted with CHCl.sub.3. The combined
extracts were washed with aq. NaHCO.sub.3 and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [500 g, n-hexane/EtOAc
(10/1)] to give methyl 3-bromo-4-nitrophenylacetate (3.69 g, 29%)
as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.68 (s, 2H),
3.73 (s, 3H), 7.38 (dd, J=8.3, 1.2 Hz, 1H), 7.67 (d, J=1.3 Hz, 1H),
7.83 (d, J=8.3 Hz, 1H).
[0686] A mixture of methyl 3-bromo-4-nitrophenylacetate (14.8 g,
53.8 mmol), reduced iron powder (9.62 g, 172 mmol), AcONa.3H.sub.2O
(7.32 g, 53.8 mmol) and AcOH (20.0 ml) in MeOH/H.sub.2O (150/600
ml) was heated at 90.degree. C. for 1 h. After cooled to room
temperature, the reaction mixture was filtered through Celite and
the filtered cake was washed with MeOH, The combined filtrate were
evaporated and extracted with EtOAc. The extracts were washed with
brine, dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was chromatographed on silica gel [400 g, CHCl.sub.3/EtOAc
(20/1)] to give methyl 4-amino-3-bromophenylacetate (9.01 g, 69%)
as a brown oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.48 (s, 2H), 3.68
(s, 3H), 4.05 (br, 2H), 6.69 (d, J=8.3 Hz, 1H), 7.00 (dd, J=8.1,
2.0 Hz, 1H), 7.32 (d, J=2.0 Hz, 1H).
[0687] To a mixture of methyl 4-amino-3-bromophenylacetate (587 mg,
2.40 mmol) and 2-methylphenyl isocyanate (0.287 ml, 2.40 mmol) in
THF (2 ml) was added Et.sub.3N (33 ml, 0.24 mmol) at room
temperature. After 21 h stirring, the reaction mixture was
concentrated in vacuo. The residue was triturated by the addition
of n-hexane to give methyl
3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetate (650 mg, 72%) as
a pale brown powder. .sup.1H-NMR (CDCl.sub.3) .delta.2.34 (s, 3H),
3.53 (s, 2H), 3.68 (s, 3H), 6.18 (br, 1H), 6.96 (br, 1H), 7.18-7.33
(m, 4H), 7.29 (d, J=4.4 Hz, 1H), 7.30 (d, J=7.3 Hz, 1H), 8.19 (d,
J=8.3 Hz, 1H); MS (ESI) m/z 377 (M.sup.+), 379 (M.sup.++2).
[0688] To a stirred solution of methyl
3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetate (650 mg, 1.72
mmol) in THF (10 ml) was added 0.25 N NaOH (10 ml). After stirring
at room temperature for 14 h, the solvent was concentrated in
vacuo. The residue was triturated by the addition of 1 N HCl and
dried at 60.degree. C. for 2 days under a reduced pressure to give
3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.22 g,
100%) as colorless powder. .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.26
(s, 3H), 3.32 (s, 2H), 6.93 (m, 2H), 7.10-7.17 (m, 4H), 7.76 (d,
J=8.1 Hz, 2H), 8.52 (s, 1H); MS (ESI) m/z 385 (M.sup.++Na), 387
(M.sup.++2+Na).
[0689] A mixture of
3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (80 mg, 0.22
mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate (56
mg, 0.22 mmol), EDC.HCl (63 mg, 0.33 mmol), HOBT (45 mg, 0.33 mmol)
and Et.sub.3N (0.15 ml, 1.10 mmol) in DMF (1 ml) was stirred at
room temperature for 18 h. The mixture was poured into ice water
and extracted with EtOAc. The combined extracts were washed with
ice water and brine. After dried over Na.sub.2SO.sub.4, the
extracts were concentrated in vacuo. The residue was purified on
TLC [CHCl.sub.3/acetone (5/1)] to give methyl
4-[1-[3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoate (140 mg, 100%) as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta.2.30 (s, 3H), 2.55 (m, 1H), 3.56
(d, J=6.4 Hz, 2H), 3.70-3.84 (m, 3H), 3.87 (s, 3H), 3.99-4.59 (m,
3H), 5.23-5.38 (m, 1H), 6.83-6.94 (m, 2H), 6.95 (d, J=8.8 Hz, 1H),
7.07-7.26 (m, 5H), 7.36-7.63 (m, 2H), 7.94-8.15 (m, 3H); MS (ESI)
m/z 598 (M.sup.++1), 600 (M.sup.++3).
[0690] To a solution of methyl
4-[1-[3-bromo-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoate (140 mg, 0.22 mmol) in THF (10 ml)
was added 0.25 N NaOH (10 ml). After stirring at room temperature
for 14 h, the mixture was concentrated under a reduced pressure and
acidified with 1 N HCl. The precipitates were collected, washed
with water and dried under a reduced pressure to give 94 (109 mg,
85%) as a colorless powder. MW 584.43 IR (KBr) 3313, 3060, 2976,
1687, 1604, 1525, 1244 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.27 (s, 3H), 2.29 (m, 2H), 3.66 (d, J=8.1 Hz, 2H),
3.72-4.68 (m, 5H), 5.31-5.53 (m, 1H), 6.92-6.99 (m, 1H), 7.04 and
7.07 (each d, J=8.3 Hz, 2H, amide isomers), 7.11-7.21 (m, 3H), 7.48
and 7.51 (s, 1H, amide isomers), 7.75 and 7.79 (each d, J=8.1 Hz,
1H, amide isomers), 7.86-7.92 (m, 3H), 8.45 and 8.47 (each s, 1H,
amide isomers), 8.59 (s, 1H), 12.64 (s, 1H); MS (FAB) m/z 584
(M.sup.++1), 586 (M.sup.++3); Anal. Calcd for
C.sub.28H.sub.27BrFN.sub.3O.sub.5: C, 57.54; H, 4.66; N, 7.19; Br,
13.67; F, 3.25. Found: C, 57.93; H, 4.97; N, 7.04; Br, 13.35; F,
2.89.
Example 89
4-[1-[3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinyl]methoxybenzoic acid
[0691] ##STR1716##
[0692] To a mixture of methyl 4-amino-3-bromophenylacetate (587 mg,
2.40 mmol) and 2-chlorophenyl isocyanate (0.29 ml, 2.40 mmol) in
THF (2 ml) was added Et.sub.3N (33 ml, 0.24 mmol) at room
temperature. After 21 h stirring, the reaction mixture was
concentrated in vacuo. The residue was triturated by the addition
of n-hexane to give methyl
3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetate (710 mg, 74%) as
a pale brown powder. .sup.1H-NMR (CDCl.sub.3) .delta. 3.57 (s, 2H),
3.70 (s, 3H), 7.02-7.28 (m, 2H), 7.36 (d, J=6.8 Hz, 1H), 7.48 (s,
1H), 8.00-8.11 (m, 2H); MS (ESI) 7/z 397 (M.sup.+), 399
(M.sup.++2), 401 (M.sup.++4).
[0693] To a stirred solution of methyl
3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetate (710 mg, 1.79
mmol) in THF (10 ml) was added 0.25 N NaOH (10 ml). After stirring
at room temperature for 14 h, the solvent was concentrated in
vacuo. The residue was triturated by the addition of 1 N HCl and
dried at 60.degree. C. for 2 days under a reduced pressure to give
3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetic acid (643 mg,
94%) as colorless powder. .sup.1H-NMR (DMSO-d.sub.6) .delta. 3.56
(s, 2H), 7.05 (m, 1H), 7.21 (dd, J=8.6, 1.7 Hz, 1H), 7.29 (t, J=7.8
Hz, 1H), 7.46 (d, J=8.1 Hz, 1H), 7.46 (d, J=8.1 Hz, 1H), 7.53 (d,
J=1.7 Hz, 1H), 7.83 (d, J=8.3 Hz, 1H), 8.06 (d, J=7.6 Hz, 1H), 8.86
(s, 1H), 8.89 (s, 1H), 12.40 (s, 1H); MS (ESI) m/z 382 (M.sup.++1),
384 (M.sup.++3).
[0694] A mixture of
3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetic acid (384 mg,
1.00 mmol), methyl 4-[(4S)-fluoro-(2S)-pyrrolidinyl]methoxybenzoate
(253 mg, 1.00 mmol), EDC.HCl (288 mg, 1.50 mmol), HOBT (203 mg,
1.50 mmol) and Et.sub.3N (0.70 ml, 5.00 mmol) in DMF (4 ml) was
stirred at room temperature for 18 h. The mixture was poured into
ice water and extracted with EtOAc. The combined extracts were
washed with ice water and brine. After dried over Na.sub.2SO.sub.4,
the extracts were concentrated in vacuo. The residue was
chromatographed on silica gel [30 g, CHCl.sub.3/acetone (10/1)] to
give methyl
4-[1-[3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoate (640 mg, 100%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.07-2.46 (m,
2H), 2.59 (t, J=18.4 Hz, 1H), 3.57 (d, J=10.5 Hz, 2H), 3.63-4.67
(m, 7H), 5.26-5.44 (m, 1H), 6.89-6.96 (m, 3H), 7.13 (d, J=7.6 Hz,
1H), 7.1 (t, J=7.3 Hz, 1H), 7.26-7.29 (m, 2H), 7.52-7.94 (m, 4H),
8.01 (d, J=8.5 Hz, 1H), 8.09 (d, J=8.5 Hz, 1H); MS (FAB) m/z 618
(M.sup.+), 620 (M.sup.++3), 622 (M.sup.++5).
[0695] To a solution of methyl
4-[1-[3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-
-pyrrolidinyl]methoxybenzoate (640 mg, 1.00 mmol) in THF (40 ml)
was added 0.25 N NaOH (40 ml). After stirring at room temperature
for 14 h, the mixture was concentrated under a reduced pressure and
acidified with 1 N HCl. The precipitates were collected, washed
with water and dried under a reduced pressure to give 95 (522 mg,
86%) as a pale yellow powder. MW 604.85 IR (KBr) 3317, 3072, 1709,
1685, 1604, 1529, 1290 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.24-2.50 (m, 2H), 3.67 (d, J=8.3 Hz, 2H), 3.73-4.68 (m,
5H), 5.31-5.52 (m, 1H), 7.03-7.09 (m, 3H), 7.22 (dt, J=8.3, 1.7 Hz,
1H), 7.30 (d, J=7.3 Hz, 1H), 7.46 (dd, J=8.0, 1.4 Hz, 1H), 7.49 and
7.52 (each d, J=2.0 Hz, 1H, amide isomers), 7.80-7.91 (m, 3H), 8.07
(dd, J=8.3, 1.2 Hz, 1H), 8.85 and 8.86 (each s, 1H, amide isomers),
8.96 and 8.97 (each s, 1H, amide isomers), 12.62 (s, 1H); MS (FAB)
m/z 605 (M.sup.++1), 607 (M.sup.++3), 609 (M.sup.++3), 626
(M.sup.++1+Na); Anal. Calcd for
C.sub.27H.sub.24BrClFN.sub.3O.sub.5.0.8H.sub.2O: C, 52.37; H, 4.17;
N, 6.79; F, 3.07. Found: C, 52.63; H, 4.12; N, 6.62; F, 2.97.
Example 90
4-[1-[3-bromo-4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)-p-
yrrolidinyl]methoxybenzoic acid
[0696] ##STR1717##
[0697] To a mixture of methyl 4-amino-3-bromophenylacetate (587 mg,
2.40 mmol) and 2-bromophenyl isocyanate (0.30 ml, 2.40 mmol) in THF
(2 ml) was added Et.sub.3N (33 ml, 0.24 mmol) at room temperature.
After 4 h stirring, the reaction mixture was concentrated in vacuo.
The residue was triturated by the addition of n-hexane to give
methyl 3-bromo-4-[N'-(2-bromophenyl)ureido)phenylacetate (770 mg,
73%) as a pale brown powder. .sup.1H-NMR (CDCl.sub.3) .delta. 3.55
(s, 2H), 3.70 (s, 3H), 6.97 (dd, J=7.3, 1.5 Hz), 1H), 7.22 (dd,
J=8.5, 2.2 Hz, 1H), 7.29-7.33 (m, 2H), 7.48 (d, J=1.0, 2.2 Hz, 1H),
7.54 (dd, J=8.0, 1.2 Hz, 1H), 8.01 (m, 2H); MS (ESI) m/z 443
(M.sup.++1), 445 (M.sup.++3), 447 (M.sup.++5).
[0698] To a stirred solution of methyl
3-bromo-4-[N'-(2-bromophenyl)ureido]phenylacetate (770 mg, 1.74
mmol) in THF (10 ml) was added 0.25 N NaOH (10 ml). After stirring
at room temperature for 14 h, the solvent was concentrated in
vacuo. The residue was triturated by the addition of 1 N HCl and
dried at 60.degree. C. for 2 days under a reduced pressure to give
3-bromo-4-[N'-(2-bromophenyl)ureido]phenylacetic acid (702 mg, 94%)
as colorless powder. .sup.1H-NMR (DMSO-d.sub.6) .delta. 3.56 (s,
2H), 6.99 (dt, J=7.8, 1.5 Hz, 1H), 7.21 (dd, J=8.3, 1.7 Hz, 1H),
7.33 (dt, J=7.1, 1.5 Hz, 1H), 7.53 (d, J=1.7 Hz, 1H), 7.62 (dd,
J=8.1, 1.5 Hz, 1H), 7.82 (d, J=8.3 Hz, 1H), 7.93 (dd, J=8.1, 1.5
Hz, 1H), 8.82 (s, 1H), 8.86 (s, 1H), 12.39 (s, 1H); MS (ESI) m/z
428 (M.sup.++1), 430 (M.sup.++3).
[0699] A mixture of
3-bromo-4-[N'-(2-chlorophenyl)ureido]phenylacetic acid (428 mg,
1.00 mmol), methyl
4-[(2S,4S)-4-fluoro-2-pyrrolidinyl]methoxybenzoate (253 mg, 1.00
mmol), EDC.HCl (288 mg, 1.50 mmol), HOBT (203 mg, 1.50 mmol) and
Et.sub.3N (0.70 ml, 5.00 mmol) in DMF (4 ml) was stirred at room
temperature for 18 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [30 g, CHCl.sub.3/acetone (10/1)] to give methyl
4-[1-[3-bromo-4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinyl]methoxybenzoate (720 mg, 100%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.07-2.45 (m,
2H), 2.58 (m, 1H), 3.58 (d, J=9.0 Hz, 2H), 3.63-4.69 (m, 9H),
5.26-5.43 (m, 1H), 6.88-6.99 (m, 3H), 7.16 (d, J=8.3 Hz, 1H),
7.23-7.32 (m, 2H), 7.46 (dd, J=8.1, 1.5 Hz, 1H), 7.51-8.20 (m, 5H);
MS (FAB) m/z 664 (M.sup.+), 666 (M.sup.++3), 668 (M.sup.++5).
[0700] To a solution of methyl
4-[1-[3-bromo-4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(4S)-fluoro-(2S)--
pyrrolidinyl]methoxybenzoate (720 mg, 1.00 mmol) in THF (40 ml) was
added 0.25 N NaOH (40 ml). After stirring at room temperature for
14 h, the mixture was concentrated in vacuo and acidified with 1 N
HCl. The mixture was extracted with CHCl.sub.3/MeOH (10/1). The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [20 g, CHCl.sub.3/acetone
(10/1)--CHCl.sub.3/MeOH (20/1)] and triturated by the addition of
ether to give 96 (489 mg, 75%) as a colorless amorphous solid. MW
649.30 IR (KBr) 3450, 3313, 3070, 1709, 1684, 1525, 1435 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) 2.25-2.50 (m, 2H), 3.67 (d, J=8.3 Hz,
2H), 3.73-4.68 (m, 5H), 5.31-5.53 (m, 1H), 6.98-7.08 (m, 3H), 7.21
(d, J=8.2 Hz, 1H), 7.34 (t, J=8.8 Hz, 1H), 7.50 and 7.53 (each s,
1H, amide isomers), 7.62 (d, J=8.0 Hz, 1H), 7.80-7.96 (m, 4H), 8.82
(s, 1H), 8.85 and 8.86 (each s, 1H, amide isomers), 12.63 (br, 1H);
MS (FAB) m/z 650 (M.sup.++1), 652 (M.sup.++3), 654 (M.sup.++3), 672
(M.sup.++Na); Anal. Calcd for
C.sub.27H.sub.24Br.sub.2FN.sub.3O.sub.5.0.9H.sub.2O: C, 48.73; H,
3.91; N, 6.31; F, 2.85. Found: C, 48.96; H, 3.98; N, 5.92; F,
2.77.
Example 91
4-[1-[4-[N'-(2-methylphenyl)ureido]-2,3-difluorophenylacetyl]-(4S)-fluoro--
(2S)-pyrrolidinyl methoxy]benzoic acid
[0701] ##STR1718##
[0702] To a stirred solution of tert-butyl ethyl malonate (5.35 ml,
28.2 mmol) in DMF (150 ml) was added NaH (60% in oil, 3.38 g, 84.7
mmol) at rt. After 20 min, 2,3-difluoronitrobenzene (5 g, 28.2
mmol) in DMF (50 mL) was added dropwise via dropping funnel.
Following the addition, the mixture was stirred for 3 hours at rt.
The mixture was poured into ice-water and sat NH.sub.4Cl (100 mL).
The mixture was extracted with EtOAc and the combined organic layer
was washed with 1M HCl and brine, dried over MgSO.sub.4, filtered
and concentrated. The residue was dissolved to dichloromethane (20
mL), and added TFA (20 mL) at rt. The mixture was refluxed for 18
h. The mixture was evaporated in vacuo, coevaporated with toluene
(20 mL.times.2). The residue was chromatographed on silica gel
(middle pressure chromatography system: YAMAZEN YFLC-5404-FC,
linear gradient hexane-EtOAc 10:0 to 1:1, .phi.50 mm.times.300 mm,
15 mL/min) to give ethyl 2,3-difluoro-4-nitrophenylacetic acid
(5.85 g, 85%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.30 (m, 3H), 3.78 (s, 2H), 4.22 (m, 2H), 7.22 (m, 1H), 7.84 (m,
1H); MS (FAB) m/z 246 (M.sup.++1).
[0703] To a stirred solution of ethyl
2,3-difluoro-4-nitrophenylacetate (5.85 g, 23.9 mol) in EtOH (100
mL), was added SnCl.sub.2 (16.1 g, 71.6 mmol) at it. The stirring
was continued for 18 hours at reflux. After removal of the solvent,
the residue was dissolved in CHCl.sub.3 (100 mL) and poured into
ice water-4M NaOH (40 mL of 4M NaOH in 300 mL of ice-water),
extracted with CHCl.sub.3 (100 mL.times.2), dried over anhydrous
MgSO.sub.4, and concentrated under a reduced pressure. The residue
was chromatographed on silica gel (middle pressure chromatography
system YAMAZEN YFLC-5404, linear gradient of hexane-EtOAc from 9:1
to 7:3, +50 mm.times.500 mm, 15 ml/min) to give ethyl
4-amino-2,3-difluorophenylacetic acid (1.94 g, 38%) as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25 (t, J=7.3 Hz, 3H), 3.55
(d, J=1.0 Hz, 2H), 3.78 (brs, 2H), 4.15 (dd, J=7.2 Hz, 14.2 Hz,
2H), 6.49 (dt, J=1.8, 8.2 Hz, 1H), 6.78 (m, 1H); MS (FAB) m/z 216
(M.sup.++1).
[0704] To a stirred solution of ethyl
4-amino-2,3-difluorophenylacetate (323 mg, 1.5 mmol) in DMF (8 mL),
were added triethylamine (0.209 ml, 1.5 mmol) and 2-methylphenyl
isocyanate (0.372 ml, 3.0 mmol) at rt. The stirring was continued
for 48 hour at 80.degree. C. The reaction mixture was evaporated in
vacuo, and the solid was suspended to n-hexane. The solid was
collected through filtration. The solid was dissolved in THF-MeOH
(1:1, v/v, 20 mL), and was added 4M NaOH (10 mL) at rt. The
stirring was continued for 18 hours at rt. The reaction was poured
into 1M HCl, and the resulting precipitate was collected through
filtration. The solid was recrystallized with CHCl.sub.3-n-hexane
to give 4-[(2-methylphenyl)ureido]-2,3-difluorophenylacetic acid
(200 mg, 42%) as a white solid. .sup.1H-NMR (CDCl.sub.3) .delta.
2.30 (s, 3H), 3.35 (s, 2H), 6.98 (m, 1H), 7.04 (m, 1H), 7.18 (d,
J=7.3 Hz, 2H), 7.69 (d, J=8.1 Hz, 1H), 7.90 (m, 1H); MS (FAB) t/z
321 (M.sup.++1).
[0705] To a stirred solution of methyl
4-(4-S-4-fluoro-(2-pyrrolidinyl)methoxy benzoate (63 mg, 0.25 mmol)
and 4-[N'-(2-methylphenyl)ureido]-2,3-difluorophenylacetic acid (82
mg, 0.25 mmol) in DMF (5 mL), were added EDC.HCl (72 mg, 0.38
mmol), HOBt (69 mg, 0.48 mmol), and DMAP (cat.), and the stirring
was continued overnight at rt. The mixture was diluted with EtOAc
(50 mL), washed with 1M NaOH, 1M HCl, and brine, dried over
anhydrous MgSO.sub.4, and concentrated under a reduced pressure.
The residue was dissolved in THF-MeOH--H.sub.2O (21 mL, 1:1:1,
v/v/v) and the stirring was continued for 6 h at nt. The mixture
was poured into 1M HCl and extracted with CHCl.sub.3-MeOH (9:1,
v/v). The combined organic phase was dried over anhydrous
MgSO.sub.4, and concentrated under a reduced pressure. The residue
was purified with TLC (Whatman, PLK-5F, CHCl.sub.3/MeOH, 20:1, v/v)
to give 97 (69 mg, 51%) as a white powder. MW 541.52 IR (KBr) 3340,
1604, 1540, 1251, 1168, 754 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.25 (s, 3H), 2.32 (m, 2H), 3.68-4.40 (m, 7H), 5.32-5.55
(m, 1H), 6.98 (m, 2H), 7.05 (d, J=8.8 Hz, 2H), 7.83 (d, J=8.8 Hz,
2H), 7.82-7.92 (m, 2H), 8.40 (s, 1H), 9.14 (s, 1H); MS (ESI) m/z
564 (M.sup.++Na); Anal. Calcd for
C.sub.28H.sub.26F.sub.3N.sub.3O.sub.5.2.0H.sub.2O: C, 58.23; H,
5.24; N, 7.28. Found: C, 58.07; H, 4.84; N, 7.03.
Example 92
4-[1-[4-[N'-(2-methylphenyl)ureido]-2,5-difluorophenylacetyl]-(4S)-fluoro--
(2S)-pyrrolidinyl methoxy]benzoic acid
[0706] ##STR1719##
[0707] To a stirred solution of di-tert-butyl ethyl malonate (6.32
ml, 28.2 mmol) in DMF (150 ml), was added NaH (60% in oil, 3.38 g,
84.7 mmol) at rt. After 20 min, 2,5-difluoronitrobenzene (5 g, 28.2
mmol) in DMF (50 mL) was added dropwise via dropping funnel.
Following the addition, the mixture was stirred for 3 hours at rt.
The mixture was poured into ice-water and sat. NH.sub.4Cl (100 mL).
The mixture was extracted with EtOAc and the combined organic layer
was washed with 1M HCl and brine, dried over MgSO.sub.4, filtered
and concentrated. The residue was dissolved to dichloromethane (20
mL), and added TFA (20 mL) at rt The mixture was refluxed for 18 h.
The mixture was evaporated in vacuo, coevaporated with toluene (20
mL.times.2). The residue was dissolved in MeOH (150 mL), and added
conc. H.sub.2SO.sub.4 (5 mL). The mixture was refluxed for 18 h.
The mixture was diluted with EtOAc (300 mL), washed with water, 1M
HCl, and brine, dried over anhydrous MgSO.sub.4, and concentrated
under a reduced pressure. The residue was chromatographed on silica
gel (middle pressure chromatography system: YAMAZEN YFLC-5404-FC,
linear gradient hexane-EtOAc 10:0 to 1:1, .phi. 50 mm.times.300 mm,
15 mL/min) to give ethyl 2,5-difluoro-4-nitrophenylacetic acid
(6.53 g, 90%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
3.75 (s, 2H), 3.76 (s, 3H), 7.29 (dd, J=5.8 Hz, 10.5 Hz, 1H), 7.81
(dd, J=6.0 Hz, 8.4 Hz, 1H); MS (ESI) m/z 232 (M.sup.++1).
[0708] To a stirred solution of ethyl
2,5-difluoro-4-nitrophenylacetate (5.88 g, 25.4 mol) in EtOH (100
mL), was added SnCl.sub.2 (17.2 g, 76.3 mmol) at rt. The stirring
was continued for 18 hours at reflux. After removal of the solvent,
the residue was dissolved in CHCl.sub.3 (100 mL) and poured into
ice water-4M NaOH (40 mL of 4M NaOH in 300 mL of ice-water),
extracted with CHCl.sub.3 (100 mL.times.2), dried over anhydrous
MgSO.sub.4, and concentrated under a reduced pressure. The residue
was chromatographed on silica gel (middle pressure chromatography
system YAMAZEN YFLC-5404, linear gradient of hexane-EtOAc from 9:1
to 7:3, +50 mm.times.500 mm, 15 ml/min) to give ethyl
4-amino-2,5-difluorophenylacetic acid (2.85 g, 52%) as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.28 (t, J=7.3 Hz, 3H), 3.51
(s, 2H), 3.78 (brs, 2H), 4.15 (dd, J=7.2 Hz, 14.2 Hz, 2H), 6.47
(dd, J=7.5, 10.4 Hz, 1H), 6.88 (dd, J=6.7, 11.0 Hz, 1H); MS (FAB)
m/z 216 (M.sup.++1).
[0709] To a stirred solution of ethyl
4-amino-2,5-difluorophenylacetate (323 mg, 1.5 mmol) in DMF (8 mL),
were added triethylamine (0.209 ml, 1.5 mmol) and 2-methylphenyl
isocyanate (0.372 ml, 3.0 mmol) at rt. The stirring was continued
for 48 hour at 80.degree. C. The reaction mixture was evaporated in
vacuo, and the solid was suspended to n-hexane. The solid was
collected through filtration. The solid was dissolved in THF-MeOH
(1:1, v/v, 20 mL), and was added 4M NaOH (10 mL) at rt. The
stirring was continued for 18 hours at rt. The reaction was poured
into 1M HCl, and the resulting precipitate was collected through
filtration. The solid was recrystallized with CHCl.sub.3-n-hexane
to give 4-[(2-methylphenyl)ureido]-2,5-difluorophenylacetic acid
(214 mg, 46%) as a white solid. .sup.1H-NMR (CDCl.sub.3) .delta.
2.30 (s, 3H), 3.35 (m, 2H), 7.02 (m, 2H), 7.18 (d, J=7.3 Hz, 2H),
7.69 (d, J=7.8 Hz, 1H), 8.03 (m, 1H); MS (FAB) m/z 321
(M.sup.++1).
[0710] To a stirred solution of methyl
4-[(4S)-fluoro-(2S)-pyrrolidinylmethoxybenzoate (63 mg, 0.25 mmol)
and 2,5-difluoro-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (82
mg, 0.25 mmol) in DMF (5 mL) was added EDC.HCl (72 mg, 0.38 mmol),
HOBt (69 mg, 0.48 mmol), and DMAP (cat.), and the stirring was
continued overnight at rt. The mixture was diluted with EtOAc (50
mL), washed with 1M NaOH, 1M HCl, and brine, dried over anhydrous
MgSO.sub.4, and concentrated under a reduced pressure. The residue
was dissolved in THF-MeOH--H.sub.2O (21 mL, 1:1:1, v/v/v) and the
stirring was continued for 6 h at rt. The mixture was poured into
1M HCl and extracted with CHCl.sub.3-MeOH (9:1, v/v). The combined
organic phase was dried over anhydrous MgSO.sub.4, and concentrated
under a reduced pressure. The residue was purified with TLC
(Whatman, PLK-5F, CHCl.sub.3/MeOH, 20:1, v/v) to give 98 (69 mg,
51%) as a white powder. MW 541.52 IR-ATR: 3351, 1604, 1537, 1167,
754 (cm.sup.-1); .sup.1H-NMR (DMSO) .delta. 2.25 (s, 3H), 2.32 (m,
2H), 3.68-4.70 (m, 7H), 5.32-5.55 (m, 1H), 6.97 (t, J=7.6 Hz, 1H),
7.06 (d, J=8.5 Hz, 2H), 7.20 (m, 3H), 7.87 (d, J=8.8 Hz, 2H),
7.83-8.04 (m, 2H), 8.45 (s, 1H), 9.18 (s, 1H); MS (ESI) m/z 564
(M.sup.++Na); Anal. Calcd for
C.sub.28H.sub.26F.sub.3N.sub.3O.sub.5.1.75H.sub.2O: C, 58.69; H,
5.19; N, 7.33. Found: C, 58.54; H, 4.85; N, 6.98.
Example 93
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-fluoro-2-pyrr-
olidinyl]methylamino benzoic acid
[0711] ##STR1720##
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-fluoro-2-pyrro-
lidinyl]methylamino benzoic acid
[0712] ##STR1721##
[0713] To a stirred solution of methyl
1-tert-butoxycarbonyl-4-fluoropyrrolidine-2-carboxylate (1.2 g,
4.85 mmol) in MeOH (5 ml) was added 1N NaOH (5 ml) and the mixture
was stirred at room temperature for 1 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The mixture was extracted with EtOAc. The extract was
washed with water, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo to give
1-tert-butoxycarbonyl-4-fluoropyrrolidine-2-carboxylic acid (1.1 g,
quant) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47
(br s, 9H), 2.78-2.83 (br s, 3H), 4.37 (s, 2H), 6.73-6.76 (m, 3H),
7.17 (m, 1H).
[0714] To a stirred solution of
1-tert-butoxycarbonyl-4-fluoropyrrolidine-2-carboxylic acid (1.1 g,
4.7 mmol) in THF (10.0 ml) was added BH.sub.3-THF (1.0 M solution
in THF, 10.0 ml, 10.0 mmol) at 0.degree. C. After stirred at room
temperature for 1.0 h. After cooled, the mixture was concentrated
in vacuo. Water was added thereto at 0.degree. C., and extracted
with EtOAc. The extract was washed with water, then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo to give
1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinylmethanol (1.0 g,
quant) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.48
(s, 9H), 2.29-2.39 (m, 1H), 3.38-3.59 (m, 2H), 3.74-3.88 (m, 2H),
4.09-4.14 (m, 2H), 4.85 (m, 1H), 5.03 (br s, 1H), 5.16 (br s,
1H).
[0715] To a stirred solution of oxalyl chloride (0.28 ml, 2.3 mmol)
in CH.sub.2Cl.sub.2 (20.0 ml) was added DMSO (0.39 ml) at
-78.degree. C. After 5 minutes, to the mixture was added
1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinylmethanol (500 mg, 2.28
mmol) in CH.sub.2Cl.sub.2 (5.0 ml). The mixture was stirred for 30
minutes at -78.degree. C., and triethylamine (1.6 ml) was added.
The mixture was stirred for 30 minutes at -78.degree. C., and
stirred for 30 minutes at room temperature. Water was added to the
mixture, and extracted with CH.sub.2Cl.sub.2. The organic layer was
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude
product was used to the subsequent reaction without further
purification. To a stirred solution of the crude product, methyl
4-aminobenzoate (302 mg, 2.0 mmol), and AcOH (0.13 ml) in DCE (10
ml) was added NaBH(OAc).sub.3 (656 mg, 3.09 mmol) at 0.degree. C.
The reaction mixture was stirred at room temperature for 18 hr. The
mixture was concentrated in vacuo. Sat. NaHCO.sub.3 was added to
the residue, and extracted with CH.sub.2Cl.sub.2. The extract was
washed with brine, dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with EtOAc-n-hexane (1:3, v/v) as eluent to give methyl
4-(1-tert-butoxycarbonyl-4-fluoro-2-pyrrolidinyl)methylaminobenzoate
(541 mg, 77%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.55-1.59 (m, 1H), 2.16-2.27 (m, 1H), 2.89-3.03 (m, 2H),
3.19-3.28 (m, 2H), 3.69-3.73 (m, 1H), 3.84 (s, 3H), 5.15 and 5.29
(each s, total 1H), 6.55-6.58 (m, 2H), 7.84-7.86 (m, 2H).
[0716] To a stirred solution of methyl
4-(1-tert-butoxycarbonyl-4-fluoro-(2-pyrrolidinyl)methylamino
benzoate (541 mg, 1.53 mmol) in CH.sub.2Cl.sub.2 (8.0 ml) was added
TFA (4.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (151 mg, 0.6 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (201 mg,
0.6 mmol), HOBt (94 mg, 0.7 mmol), and triethylamine (167 .mu.l, 1,
1.2 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl
(173 mg, 0.9 mmol) at 0.degree. C. The reaction mixture was stirred
at room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1:2, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-fluoro-2-pyr-
rolidinyl]methylaminobenzoate (320 mg, 94%) as an amorphous solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.80-1.95 (m, 1H), 2.42-2.58 (m,
1H), 3.20-3.51 (m, 3H), 3.51-3.76 (m, 5H), 3.84 (s, 3H), 3.85-3.98
(m, 1H), 4.67-4.70 (m, 1H), 5.10 and 5.23 (s, each, total 1H), 5.50
(br s, 1H), 6.49-6.52 (m, 2H), 6.78-6.81 (m, 2H), 6.97-7.01 (m,
1H), 7.14-7.18 (m, 2H), 7.24-7.36 (m, 2H), 7.80-7.82 (m, 2H),
7.99-8.01 (m, 1H), 8.15-8.18 (m, 1H).
[0717] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-4-fluoro-2-pyrrolidinyl]methylaminobenzoate (320 mg, 0.56
mmol) in THF (5.0 ml) and MeOH (3.0 ml) was added 1N NaOH (0.8 ml,
0.8 mmol). The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 99 (280 mg, 90%) as a white
crystalline solid. MW 555.00 mp 132-136.degree. C.; IR (KBr) 3332,
2937, 1602, 1531, 1174, 752 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.00-2.40 (m, 2H), 3.50-3.90 (m, 4H), 3.75-3.85 (m, 5H),
4.27 (m, 1H), 5.23 and 5.37 (each s, total 1H), 6.51-7.03 (m, 5H),
7.25-7.29 (m, 1H), 7.41-7.44 (m, 1H), 7.64-7.68 (m, 2H), 7.92-8.10
(m, 2H), 8.87-8.94 (m, 2H); Anal. calcd for
C.sub.28H.sub.28N.sub.4O.sub.5FCl.0.6H.sub.2O: C, 59.44; H, 5.20;
N, 9.90. Found: C, 59.41; H, 5.19; N, 9.72.
[0718] To a stirred solution of methyl
4-(1-tert-butoxycarbonyl-4-fluoro-(2-pyrrolidinyl)methylamino
benzoate (541 mg, 1.53 mmol) in CH.sub.2Cl.sub.2 (8.0 ml) was added
TFA (4.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (151 mg, 0.6 mmol),
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (227 mg,
0.6 mmol), HOBt (94 mg, 0.7 mmol), and triethylamine (167 .mu.l,
1.2 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl
(173 mg, 0.9 mmol) at 0.degree. C. The reaction mixture was stirred
at room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (2:3, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido)-3-methoxyphenylacetyl]-4-fluoro-2-pyrr-
olidinyl]methylamino benzoate (280 mg, 76%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.80-1.98 (m, 1H), 1.42-1.58 (m,
1H), 3.20-3.52 (m, 3H), 3.67-3.79 (m, 5H), 3.84 (s, 3H), 3.94-3.97
(m, 1H), 4.68-4.71 (m, 1H), 5.10 and 5.23 (each s, total 1H), 5.51
(br s, 1H), 6.50-6.52 (m, 2H), 6.79-7.07 (m, 5H), 7.25-7.33 (m,
1H), 7.51-7.53 (m, 1H), 7.80-7.83 (m, 2H), 7.98-8.00 (m, 1H),
8.11-8.14 (m, 1H).
[0719] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-4-fluoro-2-pyrrolidinyl]methylaminobenzoate (280 mg, 0.46
mmol) in THF (8.0 ml) and MeOH (8.0 ml) was added 1N NaOH (2.8 ml,
2.8 mmol). The mixture was stirred at 70.degree. C. for 18 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 100 (260 mg, 95%) as a white
crystalline solid. MW 599.45 mp 131-135.degree. C.; IR (KBr) 3332,
2935, 1602, 1529, 1174 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.95-2.01 (m, 1H), 2.20-2.35 (m, 1H), 3.10-3.20 (m, 1H),
3.50-3.70 (m, 3H), 3.80-3.85 (m, 5H), 4.27 (m, 1H), 5.24 and 5.37
(each s, total 1H), 6.54-6.99 (m, 5H), 7.30-7.33 (m, 1H), 7.58-7.94
(m, 3H), 7.94-7.98 (m, 2H), 8.73 (m, 1H), 8.93 (m, 1H); Anal. calcd
for C.sub.28H.sub.28N.sub.45BrF.0.7H.sub.2O: C, 54.95; H, 4.84; N,
9.15. Found: C, 54.98; H, 4.81; N, 8.93.
Example 94
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4R)-fluoro-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0720] ##STR1722##
[0721] A mixture of methyl
4-[(4R)-fluoro-(2S)-pyrrolidinylmethoxy]benzoate (634 mg, 2.50
mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(787 mg, 2.50 mmol), EDC.HCl (718 mg, 3.75 mmol), HOBt (cat.), DMAP
(cat.) and DMF (10 ml) was stirred overnight. The mixture was
diluted with EtOAc (300 ml). The solution was washed with brine
(2.times.100 ml), dried over MgSO.sub.4, and concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-EtOAc
(4:1) as eluent to give methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4R)-fluoro-(2-
S)-pyrrolidinylmethoxy]benzoate (1.37 g, quant) as a pale yellow
viscous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.24 (s, 3H),
2.26-2.47 (m, 2H), 3.46 (s, 3H), 3.49-3.64 (m, 4H), 3.87 (s, 3H),
4.06 (dd, J=9.5, 2.0 Hz, 1H), 4.51-4.62 (m, 2H), 5.20 and 5.33 (br
s, each, total 1H), 6.63 (s, 1H), 6.72 (d, J=8.3 Hz, 1H), 6.77 (d,
J=9.0 Hz, 2H), 7.05 (t, J=7.6 Hz, 1 H), 7.16-7.20 (m, 3H), 7.53 (s,
1H), 7.63 (d, J=7.8 Hz, 1H), 7.91 (d, J=9.0 Hz, 2H), 8.07 (d, J=8.1
Hz, 1H).
[0722] A mixture of methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4R)-fluoro-(2-
S)-pyrrolidinylmethoxy]benzoate (1.37 g, 2.49 mmol), 0.25 N NaOH
(20 ml, 5.00 mmol), and THF (20 ml) was stirred for 3 days. The
mixture was poured into 1 N HCl (100 ml) and extracted with
CHCl.sub.3-MeOH (5:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1 to 4:1) to
give 101 (930 mg, 70%) as a pale yellow amorphous solid. MW 535.56
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24-2.41 (m, total 5H),
3.42-4.66 (series of m, 10H), 5.31 and 5.44 (br s, each, total 1H),
6.71-7.16 (series of m, 7H), 7.79 (d, J=8.1 Hz, 1H), 7.85-7.89 (m,
2H), 7.98-8.00 (m, 1H), 8.47 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z
536 (M.sup.++1).
Example 95
4-[(4S)-chloro-1-[3-methoxy-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(2S-
)-pyrrolidinyl]methoxybenzoic acid
[0723] ##STR1723##
[0724] To a stirred solution of methyl
4-(trans-1-tert-butoxycarbonyl-4-hydroxy-(2S)-pyrrolidinyl)
methoxybenzoate (351 mg, 1.0 mmol) and Ph.sub.3P (393 mg, 1.5 mmol)
in CHCl.sub.3 (5.0 ml) was added CCl.sub.4 (5.0 ml) at room
temperature. The reaction mixture was stirred at 50.degree. C. for
24 hr. The reaction mixture was concentrated in vacuo. The residue
was purified by column chromatography on silica gel with n-hexane
to n-hexane-EtOAc (4:1, v/v) as eluent to give methyl
4-(cis-1-tert-butoxycarbonyl-4-chloro-(2S)-pyrrolidinyl)methoxybenzoate
(340 mg, 92%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.48 (s, 9H), 2.38-2.65 (m, 2H), 3.50-3.60 (m, 1H), 3.88
(s, 3H), 3.89-4.05 (m, 1H), 4.26-4.41 (m, 4H), 6.95-6.97 (m, 2H),
7.98 (d, J=8.5 Hz, 2H).
[0725] To a stirred solution of methyl
4-(1-tert-butoxycarbonyl-(4S)-chloro-(2S)-pyrrolidinyl)
methoxybenzoate (369 mg, 1.0 mmol) in CH.sub.2Cl.sub.2 (3.0 ml) was
added TFA (3.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product (185 mg, 0.5 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (167 mg,
0.5 mmol), HOBt (68 mg, 0.5 mmol), and triethylamine (208 ml, 1.5
mmol) in THF (8.0 ml) and MeCN (8.0 ml) was added EDC.HCl (144 mg,
0.75 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:2, v/v) as eluent to give methyl
4-[(4S)-chloro-1-[3-methoxy-4-[N'-(2-chlorophenyl)ureido]phenylacetyl]-(2-
S)-pyrrolidinyl]methoxy benzoate (210 mg, 72%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 3.35-3.50 (m, 1H), 3.55-3.65 (m,
1H), 3.61-3.66 (m, 3H), 3.75 (s, 3H), 3.88 (s, 3H), 3.99-4.04 (m,
1H), 4.35-4.40 (m, 3H), 4.48-4.53 (m, 1H), 6.77-7.10 (m, 7H),
7.25-7.36 (m, 2H), 7.93-8.00 (m, 2H), 8.18 (d, J=8.0 Hz, 1H).
[0726] To a stirred solution of methyl
4-[(4S)-chloro-1-[3-methoxy-4-[N'-(2-chlorophenyl)
ureido]phenylacetyl]-(2S)-pyrrolidinyl]methoxybenzoate (210 mg,
0.35 mmol) in THF (6.0 ml) and MeOH (3.0 ml) was added 1N NaOH (0.7
ml, 0.7 mmol). The mixture was stirred at 70.degree. C. for 18 hr.
The mixture was concentrate in vacuo, water was added thereto, and
neutralized with 1 N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 102 (200 mg, 98%) as a white
crystalline solid. MW 572.44 mp 126-131.degree. C.; IR (KBr) 3330,
1685, 1604, 1533, 1438 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.15-2.25 (m, 1H), 2.58-2.63 (m, 1H), 3.58-3.78 (m, 3H),
3.83 (s, 3H), 4.13-4.42 (m, 4H), 4.73 (m, 1H), 6.75-7.45 (m, 7H),
7.86-8.10 (m, 4H), 8.90 (s, 1H), 8.95 (s, 1H); MS (FAB) m/z 572
(M.sup.++1); Anal. calcd for C.sub.28H.sub.27N.sub.3O.sub.6Cl: C,
58.75; H, 4.75; N, 7.34. Found: C, 58.93; H, 4.85; N, 7.15.
Example 96
4-[1-[4-[N'-(2-bromophenyl)ureido]]-3-methoxyphenylacetyl]-(4S)-chloro-(2S-
)-pyrrolidinyl]methoxybenzoic acid
[0727] ##STR1724##
[0728] To a stirred solution of methyl
4-[1-tert-butoxycarbonyl-(4S)-chloro-(2S)-pyrrolidinyl]methoxybenzoate
(369 mg, 1.0 mmol) in CH.sub.2Cl.sub.2 (3.0 ml) was added TFA (3.0
ml) at 0.degree. C. The reaction mixture was stirred at room
temperature for 0.5 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (185 mg, 0.5 mmol),
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (190 mg,
0.5 mmol), HOBt (68 mg, 0.5 mmol), and triethylamine (208 ml, 1.5
mmol) in THF (8.0 ml) and MeCN (8.0 ml) was added EDC.HCl (144 mg,
0.75 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:2, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-chloro-(2S-
)-pyrrolidinyl]methoxybenzoate (260 mg, 83%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 2.32-2.50 (m, 1H), 2.53-2.65 (m,
1H), 3.61-3.67 (m, 3H), 3.75 (s, 3H), 3.88 (s, 3H), 3.99-4.03 (m,
1H), 4.35-4.40 (m, 3H), 4.45-4.55 (m, 1H), 6.78-7.10 (m, 7H),
7.28-7.33 (m, 1H), 7.52 (d, J=8.0 Hz, 1H), 7.94-7.99 (m, 3H), 8.14
(d, J=8.3 Hz, 1H).
[0729] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(4S)-chloro-(2S)-pyrrolidinyl]methoxybenzoate (260 mg, 0.4
mmol) in THF (6.0 ml) and MeOH (3.0 ml) was added 1N NaOH (0.8 ml,
0.8 mmol). The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrate in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 103 (210 mg, 83%) as a white
crystalline solid. MW 616.89 mp 127-132.degree. C.; IR (KBr) 3330,
1685, 1604, 1529, 1434 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.18-2.28 (m, 1H), 2.60-2.70 (m, 1H), 3.55-3.75 (m, 3H),
3.83 (s, 3H), 4.12-4.42 (m, 4H), 4.60-4.75 (m, 1H), 6.75-7.06 (m,
5H), 7.30-7.34 (m, 1H), 7.60 (d, J=7.3 Hz, 1H), 7.86-7.94 (m, 5H),
8.75 (s, 1H), 8.94 (s, 1H); MS (FAB) m/z 616 (M.sup.+), 618
(M.sup.++2); Anal. calcd for C.sub.28H.sub.27N.sub.3O.sub.6ClBr: C,
54.52; H, 4.41; N, 6.81. Found: C, 54.98; H, 4.54; N, 6.66.
Example 97
4-[(4R)-chloro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0730] ##STR1725##
4-[(4R)-chloro-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0731] ##STR1726##
[0732] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-(4S)-hydroxy-(2S)-pyrrolidinylcarboxylate
(1.81 g, 7.34 mmol) in CCl.sub.4--CH.sub.2Cl.sub.2 (20 ml, 1:1,
v/v) was added Ph.sub.3P (3.87 mmol, 14.75 mmol) and the reaction
mixture was stirred at room temperature for 2 hr. To the mixture
was added EtOH (5 ml) and the reaction mixture was stirred at room
temperature overnight. After removal of the solvent, the residue
was purified by column chromatography on silica-gel with
n-hexane-EtOAc (3:1, v/v) as eluent to give Synthesis of methyl
1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinylcarboxylate
(1.36 g, 70%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.42 (s, 9H), 2.32-2.39 (m, 1H), 2.49-2.54 (m, 1H), 3.66-3.92
(series of s and m, total 5H), 4.44-4.55 (m, 2H); MS (FAB) m/z 264
(M.sup.++1).
[0733] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinyl carboxylate
(1.35 g, 5.12 mmol) in THF (10 ml) was added 0.5 N NaOH (10 ml) and
the reaction mixture was heated under reflux for 1.5 hr. After
cooled to room temperature, the mixture was poured into ice-1 N HCl
and the mixture was extracted with CHCl.sub.3-MeOH (9:1, v/v). The
extract was washed with brine, dried over Na.sub.2SO.sub.4 and
evaporated to give
1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinylcarboxylic
acid (1.28 g, quant.) as a colorless oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.44 (s, 9H), 2.37-2.54 (m, 2H), 3.68-3.88 (m, 2H),
4.42-4.45 (m, 2H).
[0734] To a stirred solution of
1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinylcarboxylic
acid (1.28 g, 5.13 mmol) in THF (20 ml) was added dropwise
BH.sub.3DMS (0.60 ml, 6.33 mmol) via a syringe and the reaction
mixture was stirred at room temperature for 1 hr. After removal of
the solvent, the residue was dissolved in CH.sub.2Cl.sub.2. The
solution was washed with H.sub.2O, brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (50:1,
v/v) as eluent to give 1-(tert-butoxycarbonyl)-(4R)-chloroprolinol
(0.88 g, 73%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.48 (s, 9H), 1.98 (m, 1H), 2.26-2.32 (m, 1H), 3.56-3.65 (m, 2H),
3.77 (m, 2H), 4.24 (m, 1H), 4.41-4.46 (m, 2H); MS (FAB) m/z 236
(M.sup.++1).
[0735] To a cooled (0.degree. C.), stirred solution of methyl
4-hydroxybenzoate (560 mg, 3.68 mmol),
1-(tert-butoxycarbonyl)-(4R)-chloroprolinol (870 mg, 3.69 mmol),
Ph.sub.3P (1.16 g, 4.42 mmol) in THF (15 ml) was added DIAD (870
ml, 4.42 mmol) and the reaction mixture was heated under reflux for
10 hr. After cooled to room temperature, the mixture was
evaporated. The residue was purified by column chromatography on
silica-gel with n-hexane-EtOAc (5:1, v/v) as eluent to give methyl
4-[1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinylmethoxy]benzoate
(890 mg, 65%) as a white solid mp 116-120.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.47 (s, 9H), 2.39-2.53 (m, 2H), 3.69-3.70 and
4.13-4.17 (m, total 3H), 3.88 (s, 3H), 4.30-4.41 (m, 2H), 4.50-4.55
(m, 1H), 6.90-6.92 (m, 2H), 7.96-7.98 (m, 2H); MS (FAB) m/z 370
(M.sup.++1); Anal. Calcd for C.sub.18H.sub.24ClNO.sub.5: C, 58.46;
H, 6.54; Cl, 9.59; N, 3.79. Found: C, 58.35; H, 6.56; Cl, 9.75; N,
3.77.
[0736] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-(4R)-chloro-(2S)-pyrrolidinylmethoxy]benzoate
(840 mg, 2.27 mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (10
ml) and the reaction mixture was stirred at room temperature
overnight. The mixture was concentrated in vacuo and made basic by
sat. NaHCO.sub.3. The mixture was extracted with CHCl.sub.3, washed
with brine, dried over Na.sub.2SO.sub.4, and evaporated to give
methyl 4-[(4R)-chloro-(2S)-pyrrolidinylmethoxy]benzoate (580 mg,
95%) as a white solid. mp 61-64.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.85 (br s, 1H), 2.03-2.10 (m, 1H), 2.29-2.35
(m, 1H), 3.19-3.31 (m, 2H), 3.88 (s, 3H), 3.92-4.06 (m, 3H),
4.53-4.56 (m, 1H), 6.91 (d, J=8.8 Hz, 2H), 7.98 (d, J=8.8 Hz, 2H);
MS (FAB) m/z 270 (M.sup.++1).
[0737] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (385 mg,
1.22 mmol), methyl 4-[(4R)-chloro-(2S)-pyrrolidinylmethoxy]benzoate
(330 mg, 1.22 mmol), EDC.HCl (281 mg, 1.47 mmol), HOBt (200 mg,
1.48 mmol) and Et.sub.3N (205 ml, 1.47 mmol) in THF (10 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4 and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[(4R)-chloro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylace-
tyl]-(2S)-pyrrolidinylmethoxy]benzoate (670 mg, 97%) as a white
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.28 (s, 3H), 2.33-2.57 (m,
2H), 3.50 (s, 3H), 3.59-3.60 (m, 2H), 3.75-3.82 (m, 2H), 3.88 (s,
3H), 4.06-4.09 (m, 1H), 4.51-4.63 (m, 3H), 6.65-6.80 (m, 5H),
7.09-7.13 (m, 1H), 7.20-7.27 (m, 3H), 7.56-7.58 (m, 1H), 7.91-7.93
(m, 2H), 8.05-8.07 (m, 1H); MS (FAB) m/z 566 (M.sup.++1).
[0738] To a stirred solution of methyl
4-[(4R)-chloro-1-[3-methoxy-4-[N'-(2-methylphenyl)
ureido]phenylacetyl]-(2S)-pyrrolidinylmethoxy]benzoate (480 mg,
0.85 mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the
reaction mixture was heated under reflux for 2 hr. After cooled to
room temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected under a reduced pressure. The
crude solid was dissolved in CHCl.sub.3-MeOH and evaporated. The
residue was washed with Et.sub.2O to give 104 (355 mg, 76%) as a
white amorphous solid. MW 552.02 mp 128-132.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.25 (s, 3H), 2.29-2.46 (m, 2H), 3.57-3.73
(m, 2H), 3.78 (s, 3H), 3.81-3.99 (m, 2H), 4.11-4.31 (m, 2H),
4.43-4.45 and 4.64-4.67 (each m, total 1H), 4.83-4.85 (m, 1H),
6.71-7.17 (m, 7H), 7.78-7.80 (m, 1H), 7.87-7.91 (m, 2H), 7.99-8.01
(m, 1H), 8.47 (s, 1H), 8.56 (s, 1H), 12.66 (br s, 1H); MS (FAB) m/z
552 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.30ClN.sub.3O.sub.6.3/4H.sub.2O: C, 61.59; H, 5.61;
Cl, 6.27; N, 7.43. Found: C, 61.56; H, 5.51; Cl, 6.68; N, 7.26.
[0739] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (400 mg,
1.19 mmol), methyl 4-[(4R)-chloro-(2S)-pyrrolidinylmethoxy]benzoate
(320 mg, 1.19 mmol), EDC.HCl (275 mg, 1.43 mmol), HOBt (195 mg,
1.44 mmol) and Et.sub.3N (200 ml, 1.43 mmol) in THF (10 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1, v/v) as eluent to give methyl
4-[(4R)-chloro-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(2-
S)-pyrrolidinylmethoxy]benzoate (690 mg, 98%) as a pale yellow
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.35-2.41 (m, 1H), 2.49-2.59
(m, 1H), 3.55 (s, 3H), 3.57-3.70 (m, 2H), 3.73-3.86 (m, 2H), 3.88
(s, 3H), 4.06-4.09 (m, 1H), 4.54-4.66 (m, 3H), 6.67-6.81 (m, 4H),
6.95-6.99 (m, 1H), 7.23-7.25 (m, 1H), 7.29-7.33 (m, 1H), 7.47-7.49
(m, 2H), 7.90-7.99 (m, 3H), 8.18-8.21 (m, 1H); MS (FAB) m/z 586
(M.sup.++1).
[0740] To a stirred solution of methyl
4-[(4R)-chloro-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(2-
S)-pyrrolidinylmethoxy]benzoate (410 mg, 0.70 mmol) in THF (5 ml)
was added 0.5 N NaOH (5 ml) and the reaction mixture was heated
under reflux for 5 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl and the resulting precipitate
was collected under a reduced pressure. The crude solid was
dissolved in CHCl.sub.3-MeOH and evaporated. The residue was washed
with Et.sub.2O to give 105 (282 mg, 70%) as an amorphous solid.
[0741] MW 572.44 mp 131-136.degree. C.; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.29-2.35 (m, 1H), 2.44-2.47 (m, 1H), 3.58-3.74 (m, 2H),
3.78 (s, 3H), 3.81-3.99 (m, 2H), 4.10-4.32 (m, 2H), 4.44-4.46 and
4.66 (each m, total 1H), 4.84 (m, 1H), 6.74-7.04 (m, 5H), 7.26-7.30
(m, 1H), 7.43-7.45 (m, 1H), 7.87-7.91 (m, 2H), 7.96 (d, J=8.3 Hz,
1H), 8.09 (d, J=8.3 Hz, 1H), 8.90 (s, 1H), 8.94 (s, 1H); Anal.
Calcd for C.sub.28H.sub.27Cl.sub.2N.sub.3O.sub.6.3/4H.sub.2O: C,
57.39; H, 4.90; Cl, 11.66; N, 7.17. Found: C, 57.57; H, 4.94; Cl,
11.66; N, 6.89.
Example 98
4-[(4S)-hydroxy-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrroli-
dinyl]methoxybenzoic acid
[0742] ##STR1727##
[0743] To a stirred solution of methyl
4-[(4S)-acetoxy-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl]methoxy
benzoate (2.31 g, 5.87 mmol) in CH.sub.2Cl.sub.2 (46 ml) was added
TFA (10 ml) at room temperature. After 3.5 h stirring, the mixture
was concentrated in vacuo. The residue was diluted by the addition
of CH.sub.2Cl.sub.2 and 1 N NaOH, which were extracted with
CH.sub.2Cl.sub.2 The combined extracts were washed with brine,
dried over Na.sub.2SO.sub.4, which was concentrated in vacuo. The
residue was chromatographed on silica gel [100 g, CHCl.sub.3/MeOH
(20/1)] to give trimethyl
4-[(4S)-acetoxy(2S)-pyrrolidinyl]methoxybenzoate (1.89 mg, 100%) as
a pale purple solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.10 (s, 3H),
2.14 (m, 1H), 2.65 (m, 1H), 3.52-3.63 (m, H), 3.89 (s, 3H), 4.18
(m, 1H), 4.28 (d, J=5.9 Hz, 2H), 5.38 (m, 1H), 6.93 (d, J=8.8 Hz,
2H), 7.99 (d, J=8.8 Hz, 2H).
[0744] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (343 mg,
1.09 mmol), methyl
4-[(4S)-acetoxy-(2S)-pyrrolidinyl]methoxybenzoate (320 mg, 1.09
mmol), EDC.HCl (313 mg, 1.64 mmol), HOBT (222 mg, 1.64 mmol) and
Et.sub.3N (0.76 ml, 5.45 mmol) in DMF (7 ml) was stirred at room
temperature for 16 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [150 g, CHCl.sub.3/Acetone (5/1)], to give methyl
4-[(4S)-acetoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)
ureido]phenylacetyl]-(2S)-pyrrolidinyl]methoxybenzoate (520 mg,
81%) as a brown amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta.
2.00 (s, 3H, one of isomers), 2.03 (s, 3H, one of isomers), 2.28
(m, 5H), 3.54 (s, 1H), 3.58 (s, 2H), 3.64 (s, 1H), 3.67 and 3.69
(each s, 3H, amide isomers), 3.85 (d, J=5.4 Hz, 1H), 3.88 (s, 3H),
4.04 (t, J=9.3 Hz, 1H), 5.27-5.34 (m, 1H), 6.51 (m, 1H), 6.76-6.89
(m, 2H), 6.94 (d, J=8.1 Hz, 1H), 7.14 (m, 1H), 7.25 (m, 4H), 7.53
(d, J=8.3 Hz, 1H), 7.96 (d, J=8.0 Hz, 1H), 8.00-8.10 (m, 2H); MS
(ESI) m/z 590 (M.sup.++1).
[0745] To a solution of methyl
4-[(4S)-acetoxy-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrol-
idinyl]methoxybenzoate (520 mg, 0.882 mmol) in THF (30 ml), 0.25 N
NaOH (30 ml) was added. After stirring at room temperature for 2
days, the mixture was extracted with EtOAc. The aqueous layer was
acidified with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1).
The combined extracts were washed with brine. After dried over
Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was crystallized by the addition of CHCl.sub.3, EtOH and
ether to give 106 (68 mg, 14%) as a colorless powder. MW 533.57 mp
148-152.degree. C. (dec.); IR (KBr) 3356, 2939, 1687, 1604, 1533,
1454, 1255 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.1.95-2.09
(m, 2H), 2.25 (s, 3H), 3.59 (d, J=5.9 Hz, 2H), 3.71 (m, 1H), 3.81
and 3.85 (each s, 3H, amide isomers), 4.13-4.47 (m, 4H), 5.19 (br,
1H), 6.70-7.21 (m, 7H), 7.79 (d, J=7.9 Hz, 1H), 7.86 (d, J=8.8 Hz,
2H), 8.01 (d, J=8.3 Hz, 1H), 8.47 (s, 1H), 8.57 (s, 1H); MS (ESI)
m/z 533 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.7.1H.sub.2O: C, 63.15; H, 6.03; N,
7.62. Found: C, 63.29; H, 5.76; N, 7.46.
Example 99
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-hydroxy-(2-
S)-pyrrolidinyl]methoxybenzoic acid
[0746] ##STR1728##
[0747] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (342 mg,
1.02 mmol), methyl
4-[(2S,4S)-4-acetoxy-2-pyrrolidinyl]methoxybenzoate (300 mg, 1.02
mmol), EDC.HCl (293 mg, 1.53 mmol), HOBT (207 mg, 1.53 mmol) and
Et.sub.3N (0.71 ml, 5.10 mmol) in DMF (6 ml) was stirred at room
temperature for 15 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/Acetone (5/1)], to give methyl
4-[(4S)-acetoxy-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(-
2S)-pyrrolidinyl]methoxybenzoate (510 mg, 82%) as a pale brown
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. b 2.01 and 2.04
(each s, 3H, amide isomers), 2.17 (m, 2H), 3.56-3.66 (m, 3H), 3.61
(s, 3H), 3.88 (s, 3H), 3.89 (m, 1H), 4.07 (t, J=9.6 Hz, 1H), 4.45
(dd, J=9.2, 3.4 Hz, 1H), 4.56 (m, 1H), 5.31-5.39 (m, 1H), 6.80-7.01
(m, 4H), 7.23 (d, J=8.1 Hz, 4H), 7.34 (d, J=8.1 Hz, 1H), 7.95 (d,
J=8.5 Hz, 1H), 8.00 (m, 1H), 8.18 (d, J=8.3 Hz, 1H); MS (ESI) m/z
610 (M.sup.++1), 612 (M.sup.++3).
[0748] To a solution of methyl
4-[(4S)-acetoxy-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(-
2S)-pyrrolidinyl]methoxybenzoate (510 mg, 0.836 mmol) in THF (30
ml), 0.25 N NaOH (30 ml) was added. After stirring at room
temperature for 2 days, the mixture was extracted with EtOAc. The
aqueous layer was acidified with 1 N HCl and extracted with
CHCl.sub.3-MeOH (10/1). The combined extracts were washed with
brine. After dried over Na.sub.2SO.sub.4, the extracts were
concentrated in vacuo. The residue was crystallized by the addition
of EtOH and ether to give 107 (22 mg, 5%) as a colorless powder. MW
553.99 mp 138-142.degree. C. (dec.); IR (KBr) 3334, 2939, 1685,
1604, 1533, 1439, 1248 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.93-2.14 (m, 2H), 3.60 (d, J=5.7 Hz, 2H), 3.71 (m, 1H),
3.81 and 3.85 (each s, 3H, amide isomers), 4.14-4.50 (m, 4H), 5.19
(br, 1H), 6.72 and 6.76 (each m, 1H, amideisomers), 6.85 and 6.90
(each s, 1H, amide isomers), 7.00-7.08 (m, 3H), 7.28 (t, J=7.3 Hz,
1H), 7.43 (dd, J=8.1, 1.2 Hz, 1H), 7.86-7.95 (m, 2H), 7.97 (d,
J=8.1 Hz, 1H), 8.10 (dd, J=8.3, 1.5 Hz, 1H), 8.90 (s, 1H), 8.94 (s,
1H), 12.64 (br, 1H); MS (ESI) m/z 554 (M.sup.++1), 556 (M.sup.++3);
Anal. Calcd for C.sub.28H.sub.28ClN.sub.3O.sub.7: C, 60.71; H,
5.05; Cl, 6.40; N, 7.58. Found: C, 60.47; H, 5.37; Cl, 6.31; N,
7.19.
Example 100
4-[(4S)-acetoxy-1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(2S)-pyrrolid-
inyl]methoxybenzoic acid
[0749] ##STR1729##
[0750] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (387 mg,
1.02 mmol), methyl
4-[(4S)-acetoxy-(2S)-pyrrolidinyl]methoxybenzoate (300 mg, 1.02
mmol), EDC.HCl (293 mg, 1.53 mmol), HOBT (207 mg, 1.53 mmol) and
Et.sub.3N (0.71 ml, 5.10 mmol) in DMF (6 ml) was stirred at room
temperature for 15 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/Acetone (5/1)], to give methyl
4-[(4S)-acetoxy-1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(2S)-pyrroli-
dinyl]methoxybenzoate (510 mg, 76%) as a yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.01 and 2.04 (each s, 3H, amide isomers),
2.31 (m, 2H), 3.54-3.68 (m, 3H), 3.76 (s, 2H), 3.88 (s, 3H),
3.89-4.58 (m, 4H), 5.31-5.36 (m, 1H), 6.81-6.96 (m, 5H), 7.19-7.32
(m, 3H), 7.51 (d, J=8.0 Hz, 1H), 7.93-8.00 (m, 3H), 8.13 (d, J=8.3
Hz, 1H); MS (ESI) m/z 654 (M.sup.++1), 656 (M.sup.++3).
[0751] To a solution of methyl
4-[(4S)-acetoxy-1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(2S)-pyrroli-
dinyl]methoxybenzoate (510 mg, 0.779 mmol) in THF (30 ml), 0.25 N
NaOH (30 ml) was added. After stirring at room temperature for 2
days, the mixture was extracted with EtOAc. The remaining aqueous
layer was acidified with 1 N HCl and extracted with CHCl.sub.3-MeOH
(10/1). The combined extracts were washed with brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was crystallized by the addition of EtOH and ether, to give
108 (87 mg, 19%) as a pale brown powder. MW 598.44 mp
143-151.degree. C. (dec.); IR (KBr) 3332, 2937, 1685, 1604, 1529,
1529, 1435 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.92-2.14
(m, 2H), 3.60 (d, J=5.9 Hz, 2H), 3.72 (m, 1H), 3.81 and 3.85 (each
s, 3H, amide isomers), 4.14-4.49 (m, 4H), 5.19 (br, 1H), 6.72 and
6.75 (each m, 1H, amide isomers), 6.85 and 6.90 (each m, 1H, amide
isomers), 6.97 (t, J=6.1 Hz, 1H), 7.06 (d, J=8.8 Hz, 2H), 7.32 (t,
J=7.1 Hz, 1H), 7.60 (dd, J=7.8, 1.2 Hz, 1H), 7.86 (d, J=8.8 Hz,
2H), 7.87-7.97 (m, 3H), 8.74 (s, 1H), 8.93 (s, 1H), 12.60 (br, 1H);
MS (ESI) m/z 559 (M.sup.++1), 561 (M.sup.++3); Anal. Calcd for
C.sub.28H.sub.28BrN.sub.3O.sub.7.0.1H.sub.2O: C, 56.03; H, 4.74;
Br, 13.31; N, 7.00. Found: C, 55.80; H, 4.84; Br, 13.64; N,
6.66.
Example 101
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-hydroxy-(2-
S)-pyrrolidinyl methoxy]benzoic acid
[0752] ##STR1730##
[0753] To a stirred solution of methyl
4-[(4R)-acetoxy-1-(tert-butoxycarbonyl)-(2S)-pyrrolidinylmethoxy]benzoate
(835 mg, 2.12 mmol) in CH.sub.2Cl.sub.2 (5 ml) was added TFA (5 ml)
and the reaction mixture was stirred at room temperature for 1 hr.
The mixture was concentrated in vacuo and made basic by sat.
NaHCO.sub.3. The mixture was extracted with CHCl.sub.3, washed with
brine, dried over K.sub.2CO.sub.3 and evaporated to give methyl
4-[(4R)-acetoxy-(2S)-pyrrolidinylmethoxy]benzoate (580 mg, 95%) as
a brown oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.86-1.93 (m, 1H),
2.00-2.12 (series of s and m, total 5H), 3.03-3.29 (m, 1H),
3.73-3.80 (m, 1H), 3.88 (s, 3H), 3.93-4.01 (m, 2H), 5.27-5.30 (m,
1H), 6.91 (d, J=9.0 Hz, 2H), 7.98 (d, J=9.0 Hz, 2H); MS (FAB) m/z
294 (M.sup.++1).
[0754] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (365 mg,
1.09 mmol), methyl
4-[(4R)-acetoxy-(2S)-pyrrolidinylmethoxy]benzoate (320 mg, 1.09
mmol), EDC.HCl (250 mg, 1.30 mmol), HOBt (180 mg, 1.33 mmol) and
Et.sub.3N (182 ml, 1.31 mmol) in THF (5 ml) was stirred at room
temperature for 2 days. The mixture was diluted with H.sub.2O and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as
eluent to give methyl
4-[(4R)-acetoxy-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-(2S)-pyrrolidinylmethoxy]benzoate (500 mg, 75%) as a white
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.01 (s, 3H), 2.03-2.05 (m,
1H), 2.20-2.26 (m, 1H), 2.37-2.43 (m, 1H), 3.59 (s, 2H), 3.62 (s,
3H), 3.66-3.87 (m, 2H), 3.89 (s, 3H), 4.07-4.09 (m, 1H), 4.48-4.51
(m, 1H), 4.59 (m, 1H), 6.70-6.82 (m, 4H), 6.97-7.01 (m, 1H),
7.24-7.35 (m, 4H), 7.92-7.98 (m, 3H), 8.12-8.21 (m, 1H); MS (FAB)
m/z 610 (M.sup.++1).
[0755] To a stirred solution of methyl
4-[(4R)-acetoxy-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(-
2S)-pyrrolidinylmethoxy]benzoate (500 mg, 0.82 mmol) in THF (5 ml)
was added 0.5 N NaOH (5 ml) and the reaction mixture was heated
under reflux overnight. After cooled to room temperature, the
mixture was poured into ice-1 N HCl and the resulting precipitate
was collected under a reduced pressure. The crude solid was
purified by recrystallization from CHCl.sub.3--IPE to give 109 (223
mg, 49%) as a white crystalline powder. MW 553.99 mp
137-142.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.95-2.09
(m, 2H), 3.41-3.43 (m, 1H), 3.57 (m, 3H), 3.78 (s, 3H), 4.07-4.40
(series of m, total 4H), 5.07 (m, 1H), 6.72-6.74 (m, 1H), 6.85 (m,
1H), 6.99-7.03 (m, 3H), 7.25-7.29 (m, 1H), 7.42-7.43 (m, 1H),
7.85-7.87 (m, 2H), 7.93-7.95 (m, 1H), 8.07-8.09 (m, 1H), 8.88 (s,
1H), 8.92 (s, 1H), 12.65 (br s, 1H); MS (FAB) m/z 554 (M.sup.++1);
Anal. Calcd for C.sub.28H.sub.28ClN.sub.3O.sub.7.1/2H.sub.2O: C,
59.73; H, 5.19; Cl, 6.30; N, 7.46. Found: C, 59.58; H, 5.32; Cl,
6.99; N, 7.21.
Example 102
4-[(4R)-hydroxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2-
S)-pyrrolidinyl methoxybenzoic acid
[0756] ##STR1731##
[0757] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (320 mg,
1.02 mmol), methyl
4-[(4R)-acetoxy-(2S)-pyrrolidinylmethoxy]benzoate (300 mg, 1.02
mmol), EDC.HCl (235 mg, 1.23 mmol), HOBt (166 mg, 1.23 mmol) and
Et.sub.3N (171 ml, 1.23 mmol) in THF (5 ml) was stirred at room
temperature for 2 days. The mixture was diluted with H.sub.2O and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (50:1,
v/v) as eluent to give methyl
4-[(4R)-acetoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(-
2S)-pyrrolidinylmethoxy]benzoate (420 mg, 70%) as a white foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.99 (s, 3H), 2.02-2.05 (m, 1H),
2.15-2.41 (series of s and m, total 5H), 3.55 (s, 3H), 3.57 (s,
2H), 3.63-3.73 (m, 2H), 3.89 (s, 3H), 4.07-4.10 (m, 1H), 4.45-4.48
(m, 1H), 4.57 (m, 1H), 6.56 (s, 1H), 6.66 (m, 1H), 6.75-6.82 (m,
3H), 7.11-7.24 (m, 4H), 7.54-7.56 (m, 1H), 7.92-7.94 (m, 2H),
8.05-8.07 (m, 1H); MS (FAB) m/z 590 (M.sup.++1).
[0758] To a stirred solution of methyl
4-[(4R)-acetoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(-
2S)-pyrrolidinylmethoxy]benzoate (420 mg, 0.71 mmol) in THF (5 ml)
was added 0.5 N NaOH (5 ml) and the reaction mixture was heated
under reflux overnight. After cooled to room temperature, the
mixture was poured into ice-1 N HCl and the resulting precipitate
was collected under a reduced pressure. The crude solid was
purified by recrystallization from CHCl.sub.3--IPE to give 110 (182
mg, 48%) as a white crystalline powder. MW 533.57 mp
178-182.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.92-2.10
(m, 2H), 2.23 (s, 3H), 3.40-3.44 (m, 1H), 3.56-3.67 (m, 3H), 3.78
(s, 3H), 4.05-4.39 (series of m, total 4H), 5.06 (m, 1H), 6.71-7.01
(m, 5H), 7.10-7.16 (m, 2 H), 7.77-7.79 (m, 1H), 7.85-7.89 (m, 2H),
7.98-8.00 (m, 1H), 8.45 (s, 1H), 8.54 (s, 1H), 12.59 (br s, 1H); MS
(FAB) m/z 534 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.7.1/2H.sub.2O: C, 64.20; H, 5.94; N,
7.74. Found: C, 64.35; H, 5.83; N, 7.68.
Example 103
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-p-
yrrolidinylmethyl]-1-piperazinylacetic acid
[0759] ##STR1732##
[0760] To a stirred solution of N-(tert-butoxycarbonyl)
(4S)-fluoroprolinol (1.26 g, 5.75 mmol), Et.sub.3N (4 ml, 28.5
mmol) and DMSO (4.1 ml, 57.5 mmol) in CH.sub.2Cl.sub.2 (20 ml) was
added SO.sub.3.pyridine (2.74 g, 17.2 mmol). After 5 h stirring,
the mixture was evaporated to remove CH.sub.2Cl.sub.2 and diluted
with Et.sub.2O (200 ml). The solution was washed with 1 N HCl (200
ml) and brine (200 ml), dried over MgSO.sub.4 and evaporated. The
resulting residue was chromatographed on silica gel with
hexane-EtOAc (4:1) to give N-(tert-butoxycarbonyl)
(4S)-fluoroprolinal (628 mg, 50%) as a yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta.1.41-1.47 (m, 9H), 2.02-2.48 (m, 2H), 3.47-3.94
(m, 2H), 4.16 and 4.29 (each d, each J=9.8 Hz, total 1H), 5.13 and
5.26 (each s, total 1H).
[0761] To a stirred solution of N-(tert-butoxycarbonyl)
(4S)-fluoroprolinal (1.44 g, 6.63 mmol), ethyl 1-piperazinylacetate
(1.71 g, 9.94 mmol) and AcOH (759 ul, 13.3 mmol) in MeOH (20 ml)
was added NaBH.sub.3CN (880 mg, 13.3 mmol). The reaction mixture
was stirred overnight and evaporated. The residue was quenched with
sat. NaHCO.sub.3 (100 ml) and evaporated with CHCl.sub.3
(2.times.200 ml). The combined extracts were dried over MgSO.sub.4
and evaporated. The oily residue was chromatographed on silica gel
with CHCl.sub.3-MeOH (20:1) to give ethyl
4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-2-pyrrolidinylmethyl]-1-piperaziny-
lacetate (2.38 g, 95%) as a yellow oil.
[0762] A mixture of ethyl
4-[1-(tert-butoxycarbonyl)-(4S)-fluoro-2-pyrrolidinylmethyl]-1-piperaziny-
lacetate (2.38 g, 6.37 mmol), TFA (5 ml) and CH.sub.2Cl.sub.2 (5
ml) was stirred for 3 h. The mixture was evaporated and the residue
was made basic with sat. NaHCO.sub.3 (100 ml). The mixture was
extracted with CHCl.sub.3-MeOH (4:1, 2.times.150 ml) and the
combined extracts were dried over K.sub.2CO.sub.3 and evaporated to
give ethyl
4-[(4S)-fluoro-2-pyrrolidinylmethyl]-1-piperazinylacetate (1.44 g,
83%) as a brown oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.27 (dt,
J=7.1, 2.0 Hz, 3H), 1.66-3.35 (series of m, 17H), 4.18 (dq, J=7.1,
2.0 Hz, 2H), 5.09 and 5.22 (each m, total 1H).
[0763] A mixture of ethyl
4-[(4S)-fluoro-2-pyrrolidinylmethyl]-1-piperazinylacetate (1.44 g,
5.27 mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (1.66 g, 5.27 mmol), EDC.HCl (1.52 g, 7.91 mmol), HOBt (cat.)
and DMAP (cat.) in DMF (10 ml) was stirred overnight. The mixture
was diluted with EtOAc-MeOH (10:1, 220 ml). The solution was washed
with brine (200 ml), dried over MgSO.sub.4 and evaporated. The
residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(20:1) as eluent to give ethyl
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2--
pyrrolidinylmethyl]-1-piperazinylacetate (2.47 g, 82%) as a yellow
viscous solid. .sup.1H-NMR (CDCl.sub.3) 1.24-1.29 (m, 3H),
1.92-4.36 (series of m, 7H), 5.16 and 5.29 (each m, total 1H), 6.43
(s, 1H), 6.74-6.81 (m, 2H), 7.12-7.29 (m, 4H), 7.50 (d, J=7.8 Hz,
1H), 8.01-8.07 (m, 1H).
[0764] A mixture of ethyl
4-[(4S)-fluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2--
pyrrolidinylmethyl]-1-piperazinylacetate (1.0 g, 1.76 mmol) and
0.25 N NaOH (14 ml, 3.50 mmol) in THF (15 ml) was stirred
overnight. The mixture was neutralized with 1 N HCl and evaporated.
The residue was purified by ion exchange resin (DIAION, HP20) with
H.sub.2O to MeOH as eluent to give 111 MW 541.61 (400 mg, 40%) as a
pale yellow amorphous solid. .sup.1H-NMR (CD.sub.3OD) .delta.
2.00-3.95 (series of m, 24H), 4.34-4.40 (m, 1H), 5.23 and 5.36 (m,
each, total 1H), 6.78-6.82 (m, 1H), 6.92 (m, 1H), 7.00-7.04 (m,
1H), 7.09-7.23 (m, 4H), 7.59 (d, J=7.1 Hz, 1H), 7.99-8.02 (m, 1H);
MS (FAB) m/z 542 (M.sup.++1).
Example 104
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4,4-difluoro-2--
pyrrolidinylmethyl]-1-piperazinylacetic acid
[0765] ##STR1733##
[0766] To a stirred mixture of
1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethanol (2.11
g, 8.89 mmol), Et.sub.3N (6.2 ml, 44.5 mmol), DMSO (6.3 ml, 88.9
mmol) in CH.sub.2Cl.sub.2 (20 ml) was added SO.sub.3-pyridine (4.25
g, 26.7 mmol). After 3 h stirring, the mixture was concentrated in
vacuo and diluted with Et.sub.2O (200 ml). The resulting mixture
was washed with 1 N HCl (100 ml) and brine (100 ml), dried over
MgSO.sub.4, and concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-EtOAc (4:1) as eluent to
give 1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinecarbaldehyde
(1.40 g, 67%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.45-1.52 (m, 9H), 2.49 (m, 2H), 3.75-3.88 (m, 2H), 4.29-4.42 (m,
1H), 9.54 and 9.60 (s, each, total 1H).
[0767] To a stirred solution of
1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinecarbaldehyde
(1.40 g, 5.95 mmol) and ethyl 1-piperazinylacetate (1.02 g, 5.95
mmol) in MeOH--AcOH (12:1, 13 ml) was added NaBH.sub.3CN (787 mg,
11.9 mmol) at 0.degree. C. After 3 days stirring, the mixture was
quenched by addition of sat. NaHCO.sub.3 (100 ml) and extracted
with CHCl.sub.3 (2.times.200 ml). The combined extracts were dried
over MgSO.sub.4 and concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give ethyl
4-[1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethyl]-1-piperazin-
ylacetate (822 mg, 35%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.27 (t, J=7.1 Hz, 3H), 1.46 (m, 9H), 1.64 (m, 2H),
2.39-2.64 (m, 10H), 3.19 (s, 2H), 3.42-4.05 (series of m, 3H), 4.18
(q, J=7.1 Hz, 2H).
[0768] A solution of ethyl
4-[1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethyl]-1-piperazin-
ylacetate (820 mg, 2.09 mmol) and TFA (5 ml) in CH.sub.2Cl.sub.2 (5
ml) was stirred for 1 h. The mixture was concentrated in vacuo and
the residue was made basic with sat. NaHCO.sub.3. The resulting
mixture was extracted with CHCl.sub.3-MeOH (5:1, 2.times.200 ml).
The combined extracts were dried over K.sub.2CO.sub.3 and
concentrated in vacuo to give ethyl 4-(4,4-difluoro-2-pyrrolidinyl
methyl)-1-piperazinylacetate (493 mg, 81%) as a brown oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.27 (t, J=7.1 Hz, 3H), 1.91 (m,
2H), 2.27-2.60 (m, 10H), 3.09-3.34 (m, 4H), 3.46-3.53 (m, 1H), 4.19
(q, J=7.1 Hz, 2H).
[0769] A mixture of ethyl
4-(4,4-difluoro-2-pyrrolidinylmethyl)-1-piperazinylacetate (490 mg,
1.69 mmol), 4-[N'-(2-chlorophenyl)ureido)-3-methoxyphenylacetic
acid (567 mg, 1.69 mmol), EDC.HCl (486 mg, 2.54 mmol), HOBt (cat.)
and DMAP (cat.) in DMF (10 ml) was stirred overnight. The mixture
was diluted with EtOAc (250 ml), washed with brine (2.times.200
ml), dried over MgSO.sub.4, and concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-EtOAc (4:1) to
CHCl.sub.3-MeOH (10:1) as eluent to give ethyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4,4-difluoro-2-
-pyrrolidinylmethyl]-1-piperazinylacetate (973 mg, 95%) as a yellow
viscous oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25 (t, J=7.1 Hz,
3H), 2.31-2.68 (m, 12H), 3.17-3.20 (m, 2H), 3.52-3.91 (m, 4H),
4.10-4.48 (series of m, 3H), 6.75-6.84 (m, 2H), 7.00 (dt, J=7.8,
1.5 Hz, 1H), 7.16-7.29 (m, 3H), 7.35 (dd, J=8.3, 1.5 Hz, 1H), 8.00
(d, J=8.3 Hz, 1H), 8.18 (dd, J=8.3, 1.5 Hz, 1H).
[0770] To a stirred solution of ethyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4,4-difluoro-2-
-pyrrolidinylmethyl]-1-piperazinylacetate (292 mg, 0.480 mmol) in
THF (4 ml) was added 0.25 N NaOH (3.8 ml, 0.960 mmol). After 2 days
stirring, the mixture was neutralized with 1 N HCl and extracted
with CHCl.sub.3-MeOH (4:1, 2.times.200 ml). The combined extracts
were dried over MgSO.sub.4 and concentrated in vacuo. The residue
was purified by tin layer column chromatography on silica gel with
CHCl.sub.3-MeOH (5:1) to give 112 MW 580.02 (81.7 mg, 29%) as a
pale yellow amorphous solid. MW 580.02 .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.24-2.50 (series of m, 12H), 3.40-4.47 (series of m, 10H),
6.76 (d, J=8.1 Hz, 1H), 6.88 (s, 1H), 7.02 (t, J=8.1 Hz, 1H), 7.28
(t, J=8.1 Hz, 1H), 7.44 (d, J=8.1 Hz, 1H), 7.97 (d, J=8.1 Hz, 1H),
8.08 (d, J=8.1 Hz, 1H), 8.96-8.99 (m, 2H). MS (FAB) m/z 580
(M.sup.++1).
Example 105
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-phenoxy(2S-
)-pyrrolidinyl]methyl-1-piperazinylacetic acid
[0771] ##STR1734##
[0772] To a stirred mixture of methyl
(2S,4R)-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinylcarboxylate
(4.69 g, 19.1 mmol), phenol (1.98 g, 21.0 mmol) and PPh.sub.3 (5.51
g, 21.0 mmol) in THF (80 ml) was added DIAD (4.13 ml, 21.0 mmol) at
room temperature under an atmosphere of nitrogen. The mixture was
stirred over night. After removal of the solvent, the resulting
residue was chromatographed on silica gel [700 g, CHCl.sub.3/EtOAc
(10/1)], to give methyl
(2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidinylcarboxylate
(5.31 g, 86%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.43 (br, 9H, one of isomers), 1.48 (br, 9H, one of isomers), 2.48
(m, 1H), 3.75 (br, 3H), 4.42-4.96 (m, 2H), 6.88-7.35 (m, 5H).
[0773] To a stirred solution of methyl
(2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidinylcarboxylate
(5.31 g, 16.5 mmol) in THF (132 ml) was added 0.25 N NaOH (132 ml,
33.0 mmol) at room temperature. The resulting mixture was stirred
over night After removal of the solvent, the mixture was acidified
by the addition of 1 N HCl and extracted with CHCl.sub.3. The
combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was recrystallized
from n-hexane-CHCl.sub.3, to give
(2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidinylcarboxylic
acid (2.96 g, 58%) as a white powder. .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.36 (s, 9H), 2.16 (d, J=13.2 Hz, 1H), 2.56 (m, 1H), 3.46
(m, 1H), 3.71 (dt, J=12.0, 5.4 Hz, 1H), 4.26 (dt, J=9.5, 7.1 Hz,
1H), 4.99 (m, 1H), 6.85 (m, 2H), 6.94 (t, J=7.3 Hz, 1H), 7.28 (t,
J=7.3 Hz, 1H).
[0774] To a stirred solution of
(2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidinylcarboxylic
acid (2.39 g, 7.76 mmol) in THF (50 ml) was added BH.sub.3-DMS
(1.55 ml, 15.5 mmol) at 0.degree. C. After 10 min. stirring at the
same temperature, the mixture was allowed to room temperature and
then heated at 50.degree. C. for 2 h. After cooling to room
temperature, the mixture was concentrated in vacuo and quenched by
the addition of water at 0.degree. C. The mixture was extracted
with EtOAc. The combined extracts were washed with brine, dried
over Na.sub.2SO.sub.4 and evaporated. The residue was
chromatographed on silica gel [60 g, CHCl.sub.3/MeOH (50/1)] to
give (2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidinylmethanol
(2.83 g, 100%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.47 (s, 9H), 1.95 (br, 1H), 2.36 (m, 1H), 3.56-3.74 (m, 3H),
3.89-4.52 (m, 3H), 4.85 (br, 1H), 6.84 (dd, J=8.8, 1.2 Hz, 2H),
6.97 (t, J=7.2 Hz, 1H), 7.29 (t, 2H, J=7.8 Hz).
[0775] To a stirred mixture of
(2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidinylmethanol
(2.75 g, 9.37 mmol), Et.sub.3N (7.84 ml, 56.2 mmol), DMSO (6.66 ml,
9.37 mmol) in CH.sub.2Cl.sub.2 (30 ml) at 0.degree. C. was added
SO.sub.3-pyridine (4.47 g, 28.1 mmol), then the resulting mixture
was allowed to raise to room temperature. After 2.5 h stirring, the
mixture was concentrated in vacuo. To the resulting mixture was
added water and extracted with Et.sub.2O. The combined extracts
were washed with brine, dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was chromatographed on silica
gel [100 g, CHCl.sub.3/acetone (5/1)] to give
(2S,4S)-1-tert-butoxycarbonyl-4-phenoxy-2-pyrrolidine carbaldehyde
(2.54 g, 93%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.45 (s, 9H, one of isomers), 1.49 (s, 9H, one of isomers), 2.17
(br, 2H), 3.65-4.31 (m, 3H), 4.91 (br, 1H), 6.79 (d, J=7.8 Hz, 1H),
6.77 (m, 1H), 7.28 (m, 2H), 9.66 (m, 1H).
[0776] To a stirred mixture of
1-tert-butoxycarbonyl-(4S)-phenoxy-(2S)-pyrrolidinecarbaldehyde
(1.36 g, 4.67 mmol), ethyl 1-piperazinylacetate (1.61 g, 9.37 mmol)
in THF (30 ml) was added NaBH(OAc).sub.3 (1.98 g, 9.34 mmol) at
room temperature. After 3 h stirring, the mixture was quenched by
the addition of water and extracted with EtOAc. The combined
extracts were washed with aq. NaHCO.sub.3 and brine. The organic
layer was dried over Na.sub.2SO.sub.4 and concentrated in vacuo.
The residue was chromatographed on silica gel [50 g,
CHCl.sub.3/MeOH (10/1)], to give ethyl
4-[1-tert-butoxycarbonyl-(4S)-phenoxy-(2S)-pyrrolidinyl]methyl-1-piperazi-
nylacetate (1.05 g, 50%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.27 (t, J=7.3 Hz, 3H), 1.57 (s, 9H), 2.18 (m,
1H), 2.33-2.74 (m, 9H), 3.17 (s, 2H), 3.52-4.10 (m, 5H), 4.17 (q,
J=7.3 Hz, 2H), 4.89 (br, 1H), 6.84 (d, J=6.8 Hz, 2H), 6.95 (m, 1H),
7.26 (m, 3H).
[0777] To a stirred solution of ethyl
4-[1-tert-butoxycarbonyl-(4S)-phenoxy-(2S)-pyrrolidinyl]methyl-1-piperazi-
nylacetate (1.05 g, 2.35 mmol) in CH.sub.2Cl.sub.2 (20 ml) was
added TFA (20 ml) at room temperature. After 3 h stirring, the
mixture was concentrated in vacuo, which was diluted with
CHCl.sub.3-MeOH (10/1) and made basic by the addition of 1 N NaOH,
The combined reaction mixture was extracted with CHCl.sub.3-MeOH
(10/1). The organic layer was washed with brine, dried over
NaSO.sub.4 and concentrated, to give ethyl
4-[(4S)-phenoxy-(2S)-pyrrolidinyl]methyl-1-piperazinylacetate (1.12
g, quant.) as a brown oil, which was used without further
purification. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25 (tt, J=7.1, 7.1
Hz, 3H), 1.91 (d, J=12.0 Hz, 1H), 2.42-2.85 (m, 10H), 3.22 (s, 2H),
3.50 (s, 2H), 3.54-3.82 (m, 2H), 4.15 (q, J=7.1 Hz, 2H), 4.98 (br,
1H), 6.84 (d, J=8.1 Hz, 2H), 6.76 (t, J=7.1 Hz, 1H), 7.26-7.31 (m,
3H), 7.40 (br, 1H).
[0778] A mixture of
4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetic acid (337 mg,
1.07 mmol), ethyl
4-[(4S)-phenoxy-(2S)-pyrrolidinyl]methyl-1-piperazinylacetate (373
mg, 1.07 mmol), EDC.HCl (308 mg, 1.61 mmol) and DMAP (197 mg, 1.61
mmol) in DMF (6 ml) was stirred at room temperature for 18 h. The
mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [50 g, CHCl.sub.3/MeOH
(50/1)], to give ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-phenoxy-(-
2S)-pyrrolidinyl]methyl-1-piperazinylacetate (430 mg, 62%) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25
(t, J=7.8 Hz, 3H), 1.98-2.17 (m, 2H), 2.26 (s, 3H), 2.36-2.82 (m,
1H), 3.13 (s, 1H), 3.17 (s, 1H), 3.55 (d, J=2.4 Hz, 1H), 3.66 (d,
J=0.9 Hz, 3H), 3.67-3.84 (m, 2H), 3.98-4.35 (m, 3H), 4.85-4.95 (m,
1H), 6.27-6.89 (m, 6H), 7.08 (t, J=7.3 Hz, 1H), 7.19 (d, J=6.8 Hz,
1H), 7.28 (m, 2H), 7.41 (d, J=4.9 Hz, 1H), 7.55 (d, J=7.8 Hz, 1H),
8.05 (dd, J=7.3, 2.2 Hz, 1H). For HCl salt: a pale brown amorphous
solid. IR (KBr) 3265, 3059, 1747, 1533, 1225 cm.sup.-1; MS (FAB)
m/z 644 (M.sup.++1); Anal. Calcd for
C.sub.36H.sub.45N.sub.5O.sub.6.HCl.2.1H.sub.2O: C, 60.22; H, 7.05;
N, 9.75. Found: C, 59.97; H, 6.72; N, 9.54.
[0779] To a solution of ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-phenoxy-(-
2S)-pyrrolidinyl]methyl-1-piperazinylacetate (240 mg, 0.373 mmol)
in THF (3.0 ml), 0.25 N NaOH (3.0 ml) was added. After stirring at
room temperature for 20 h, the mixture was neutralized with 1 N HCl
and extracted with CHCl.sub.3-MeOH (10/1). The combined extracts
were dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was triturated by the addition of ether, to give 113 MW
615.72 (143 mg, 62%) as a white powder. IR (KBr) 3346, 2949, 1633,
1533, 1227 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.76 (m,
1H), 2.18 (br, 2H), 2.25 (s, 3H), 2.42-2.83 (m, 9H), 3.17 (s, 1H),
3.20 (s, 1H), 3.38 (m, 1H), 3.70 (s, 2H), 3.72-3.78 (m, 2H), 3.85
(s, 3H, one of isomers), 3.87 (s, 3H, one of isomers), 3.95 (m,
1H), 4.27 (br, 1H), 5.08 (m, 1H), 6.75 (d, J=8.3 Hz, 1H), 6.93 (m,
5H), 7.14 (m, 2H), 7.30 (d, J=7.6 Hz, 1H), 7.79 (d, J=9.0 Hz, 1H),
8.02 (m, 1H), 8.50 (s, 1H), 8.58 (s, 1H); MS (FAB) m/z 616
(M.sup.++1); Anal. Calcd for
C.sub.34H.sub.41N.sub.5O.sub.6.0.1EtOH.2H.sub.2O: C, 62.58; H,
7.00; N, 10.67. Found: C, 62.73; H, 6.58; N, 10.24.
Example 106
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-phenoxy-(2-
S)-pyrrolidinyl methyl-1-piperazinylacetic acid
[0780] ##STR1735##
[0781] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (358 mg,
1.07 mmol), ethyl
4-[(4S)-phenoxy-(2S)-pyrrolidinylmethyl]-1-piperazinylacetate (373
mg, 1.07 mmol), EDC.HCl (308 mg, 1.61 mmol) and DMAP (197 mg, 1.61
mmol) in DMF (6 ml) was stirred at room temperature for 18 h. The
mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [50 g, CHCl.sub.3/MeOH
(50/1)] to give ethyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-phenoxy-(-
2S)-pyrrolidinyl]methyl-1-piperazinylacetate (320 mg, 45%) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25
(qq, J=7.3, 7.3 Hz, 3H), 2.05-2.20 (m, 2H), 2.35-2.80 (m, 1H), 3.12
and 3.18 (each s, 1H, amide isomers), 3.57 (d, J=5.4 Hz, 1H), 3.66
and 3.38 (each s, 3H, amide isomers), 3.70-3.90 (m, 2H), 4.02-4.43
(m, 3H), 4.88-4.97 (m, 1H), 6.83-6.99 (m, 6H), 7.18-7.32 (m, 3H),
7.68 (d, J=8.3 Hz, 1H), 7.74 (s, 1H), 8.16 (dd, J=8.3, 1.4 Hz,
1H).
[0782] For HCl salt: a pale brown amorphous solid. IR (KBr) 3300,
2978, 1745, 1533, 1225 cm.sup.-1; MS (FAB) m/z 664 (M.sup.+30 1),
666 (M.sup.++3); Anal. Calcd for
C.sub.35H.sub.42ClN.sub.5O.sub.6.HCl.2.4H.sub.2O: C, 56.51; H,
6.48; N, 9.41. Found: C, 56.51; H, 6.18; N, 9.28.
[0783] To a solution of ethyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-phenoxy-(-
2S)-pyrrolidinyl]methyl-1-piperazinylacetate (181 mg, 0.273 mmol)
in THF (2.2 ml), 0.25 N NaOH (2.2 ml) was added. After stirring at
room temperature for 20 h, the mixture was neutralized with 1 N HCl
and extracted with CHCl.sub.3-MeOH (10/1). The combined extracts
were dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was triturated by the addition of ether to give 114 (133
mg, 77%) as a white powder. MW 636.14 IR (KBr) 3317, 2949, 1701,
1631, 1595, 1225 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
2.13-3.05 (m, 1H), 3.22 and 3.36 (each s, 2H, amide isomer), 3.38
(m, 1H), 3.60 (s, 2H), 3.71 (m, 1H), 3.85 (s, 3H), 3.95 (m, 1H),
4.28 (br, 1H), 5.06 (m, 1H), 6.76 (d, J=8.3 Hz, 1H), 6.91-7.03 (m,
5H), 7.29 (m, 3H), 7.44 (d, J=7.9 Hz, 1H), 7.97 (dd, J=8.1, 4.1 Hz,
1H), 8.08 (d, J=7.0 Hz, 1H), 8.91 (s, 1H), 8.95 (s, 1H); MS (FAB)
m/z 636 (M.sup.++1), 638 (M.sup.++3); Anal. Calcd for
C.sub.33H.sub.38ClN.sub.5O.sub.6.0.2EtOH.1.3H.sub.2O: C, 59.98; H,
6.30; N, 10.47. Found: C, 60.25; H, 6.12; N, 10.11.
Example 107
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-phenoxy-(2S-
)-pyrrolidinyl]methyl-1-piperazinylacetic acid
[0784] ##STR1736##
[0785] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (406 mg,
1.07 mmol), ethyl
4-[(4S)-phenoxy-(2S)-pyrrolidinyl]methyl-1-piperazinylacetate (373
mg, 1.07 mmol), EDC.HCl (308 mg, 1.61 mmol) and DMAP (197 mg, 1.61
mmol) in DMF (6 ml) was stirred at room temperature for 18 h. The
mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [50 g, CHCl.sub.3/MeOH
(50/1)] to give ethyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-phenoxy-(2-
S)-pyrrolidinyl]methyl-1-piperazinylacetate (560 mg, 74%) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25
(tt, J=7.1, 7.1 Hz, 3H), 2.04-2.84 (m, 13H), 3.12 and 3.18 (each s,
1H, amide isomers), 3.57 (d, J=4.4 Hz, 1H), 3.66 and 3.68 (each s,
3H, amide isomers), 3.68-3.87 (m, 3H), 4.05-4.41 (m, 2H), 4.87-4.96
(m, 1H), 6.76-6.98 (m, 6H), 7.20-7.33 (m, 3H), 7.46 (d, J=8.1 Hz,
1H), 7.67 and 7.71 (each s, 1H, amide isomers), 7.78 and 7.81 (each
s, 1H, amide isomers), 7.95 (d, J=8.3 Hz, 1H), 8.09 (d, J=8.1 Hz,
1H). For HCl salt: a pale brown amorphous solid. IR (KBr) 3384,
2978, 1745, 1340, 1120 cm.sup.-1; MS (FAB) m/z 708 (M.sup.++1), 710
(M.sup.++3); Anal. Calcd for
C.sub.36H.sub.42BrN.sub.5O.sub.6.HCl.2.5H.sub.2O: C, 53.20; H,
6.12; N, 8.86. Found: C, 52.98; H, 5.79; N, 8.66.
[0786] To a solution of ethyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-phenoxy-(2-
S)-pyrrolidinyl]methyl-1-piperazinylacetate (330 mg, 0.466 mmol) in
THF (3.7 ml), 0.25N NaOH (3.7 ml) was added. After stirring at room
temperature for 20 h, the mixture was neutralized with 1 N HCl and
extracted with CHCl.sub.3-MeOH (10/1). The combined extracts were
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was triturated by the addition of ether, to give 115 (217 mg, 68%)
as a white powder. MW 680.59 IR (KBr) 3315, 3095, 2941, 1631, 1529,
1435 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.75 (m, 1H),
2.18 and 2.23 (each s, 2H, amide isomers), 2.30-2.78 (m, 9H), 3.15
(2, 2H), 3.47 (m, 1H), 3.58 (s, 2H), 3.60-3.82 (m, 3H), 3.84 (m,
3H), 3.93 (m, 1H), 4.26 (br, 1H), 5.08 (m, 1H), 6.75 (d, J=7.8 Hz,
1H), 6.98 (m, 5H), 7.30 (m, 3H), 7.60 (dd, J=8.1, 2.2 Hz, 1H), 7.94
(m, 2H), 8.75 (s, 1H), 8.93 (s, 1H); MS (FAB) m/z 680 (M.sup.++1),
82 (M.sup.++3); Anal. Calcd for
C.sub.33H.sub.38BrN.sub.5O.sub.6.0.2EtOH.2H.sub.2O: C, 55.27; H,
6.00; N, 9.65. Found: C, 55.32; H, 5.56; N, 9.25.
Example 108
4-[(4S)-(4-carboxyphenoxy)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]pheny-
lacetyl]-(2S)-pyrrolidinylmethoxy]benzoic acid
[0787] ##STR1737##
[0788] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-(4R)-hydroxy-(2S)-pyrrolidinylcarboxylate
(10.4 g, 0.04 mol) and imidazole (8.66 g, 0.13 mol) in DMF (40 ml)
was added TBS-Cl (7.03 g, 0.05 mol) and the reaction mixture was
stirred at 60.degree. C. for 3 hr. After cooled to room
temperature, the mixture was diluted with brine, and extracted with
Et.sub.2O. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with n-hexane-EtOAc (5:1, v/v)
as eluent to give methyl
1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-pyrrolidin-
ylcarboxylate (15.0 g, 98%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 0.06 (s, 6H), 0.87 (s, 9H), 1.41 and 1.46
(each s, 9H), 1.99-2.03 (m, 1H), 2.16-2.18 (m, 1H), 3.31-3.42 (m,
1H), 3.56-3.63 (m, 1H), 3.73 and 3.74 (each s, 3H), 4.31-4.42 (m,
2H); MS (ESI) m/z 360 (M.sup.++1).
[0789] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-pyrrolidin-
ylcarboxylate (15.0 g, 0.04 mol) in THF (60 ml) was added 1 N NaOH
(60 ml) and the reaction mixture was stirred at 60.degree. C. for 2
hr. After cooled to room temperature, the mixture was concentrated
to a small volume, acidified with 1 N HCl, and extracted with
EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated to give
1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-pyrrolidin-
ylcarboxylic acid (12.8 g, 89%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 0.07 and 0.08 (each s, 6H), 0.87 (s, 9H), 1.49
(s, 9H), 2.06-2.11 (m, 1H), 2.41-2.44 (m, 1H), 3.40-3.59 (m, 2H),
4.36-4.50 (m, 2H); MS (ESI) m/z 346 (M.sup.++1). To a cooled
(0.degree. C.) stirred solution of
1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-pyrrolidin-
ylcarboxylic acid (12.8 g, 0.04 mol) in THF (150 ml) was added
dropwise BH.sub.3-DMS (5.30 ml, 0.06 mol) and the reaction mixture
was stirred at room temperature overnight. The reaction mixture was
quenched by sat. NH.sub.4Cl, and extracted with EtOAc. The extract
was washed with brine, dried over Na.sub.2SO.sub.4, and evaporated.
The residue was purified by column chromatography on silica-gel
with toluene-acetone (5:1, v/v) as eluent to give 1-(tert-butoxy
carbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-prolinol (10.5 g,
85%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 0.06 (s,
6H), 0.87 (s, 9H), 1.47 (s, 9H), 1.58-1.63 (m, 1H), 1.93-1.98 (m,
1H), 3.32-3.44 (m, 2H), 3.51-3.57 (m, 1H), 3.67-3.71 (m, 1H),
4.13-4.15 (m, 1H), 4.27 (m, 1H), 4.87-4.89 (m, 1H).
[0790] To a cooled (0.degree. C.), stirred solution of methyl
4-hydroxybenzoate (4.81 g, 0.03 mol),
1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-prolinol
(10.5 g, 0.03 mol), and Ph.sub.3P (9.96 g, 0.04 mol) in THF (160
ml) was added dropwise DIAD (7.48 ml, 0.04 mol) and the reaction
mixture was heated under reflux for 7 hr. After cooled to room
temperature, the mixture was evaporated. The residue was purified
by column chromatography on silica-gel with n-hexane-EtOAc (6:1,
v/v) to give methyl
4-[1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethylsilyloxy)-(2S)-pyrroli-
dinylmethoxy]benzoate (9.58 g, 65%) as a white solid. mp
86-88.degree. C.; .sup.1H-NMR (CDCl.sub.3) .delta. 0.08 (s, 6H),
0.88 (s, 9H), 1.46 (s, 9H), 2.04-2.15 (m, 2H), 3.29-3.48 (m, 2H),
3.88 (s, 3H), 4.06-4.30 (m, 3H), 4.46-4.51 (m, 1H), 6.91-6.93 (m,
2H), 7.96-7.98 (m, 2H); MS (ESI) m/z 466 (M.sup.++1).
[0791] To a cooled (0.degree. C.), stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-(4R)-(tert-butyldimethyl
silyloxy)-(2S)-pyrrolidinylmethoxy]benzoate (1.49 g, 3.20 mmol) in
THF (15 ml) was added TBAF (6.40 ml, 6.40 mmol, 1 M solution in
THF) and the reaction mixture was stirred at room temperature for 2
hr. The mixture was diluted with EtOAc, washed with H.sub.2O,
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
toluene-acetone (5:1, v/v) as eluent to give methyl
4-[1-(tert-butoxycarbonyl)-(4R)-hydroxy-(2S)-pyrrolidinylmethoxy]benzoate
(1.05 g, 93%) as a white solid. mp 103-105.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.46 (s, 9H), 2.11-2.28 (m, 2H), 3.49-3.60 (m,
2H), 3.88 (s, 3H), 4.15-4.34 (m, 3H), 4.53-4.57 (m, 1H), 6.91 (d,
J=8.6 Hz, 2H), 7.97 (d, J=8.6 Hz, 2H); MS (ESI) m/z 352
(M.sup.++1). To a cooled (0.degree. C.), stirred solution of methyl
4-hydroxybenzoate (0.56 g, 3.68 mmol), methyl
4-[1-(tert-butoxycarbonyl)-(4R)-hydroxy-(2S)-pyrrolidinylmethoxy]benzoate
(1.30 g, 3.70 mmol), and Ph.sub.3P (1.16 g, 4.42 mmol) in THF (20
ml) was added dropwise DIAD (0.87 ml, 4.42 mmol) and the reaction
mixture was stirred at room temperature for 3 hr. The mixture was
evaporated and the residue was purified by column chromatography on
silica-gel with n-hexane-EtOAc (6:1, v/v) as eluent to give methyl
4-[1-(tert-butoxycarbonyl)-(4S)-(4-methoxycarbonylphenoxy)-(2S)-pyrrolidi-
nylmethoxy]benzoate (1.80 g, q.y.) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.49 (s, 9H), 2.31-2.38 (m, 1H), 2.45-2.49 (m,
1H), 3.64-3.77 (m, 2H), 3.88 (s, 6H), 4.07-4.15 (m, 1H), 4.33-4.44
(m, 2H), 4.95-5.01 (m, 1H), 6.85 (d, J=8.8 Hz, 2H), 6.94 (br s,
2H), 7.97 (d, J=8.8 Hz, 4H); MS (ESI) m/z 486 (M.sup.++1).
[0792] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-(4S)-(4-methoxycarbonyl
phenoxy)-(2S)-pyrrolidinylmethoxy]benzoate (1.80 g, 3.71 mmol) in
CH.sub.2Cl.sub.2 (15 ml) was added TFA (15 ml) and the reaction
mixture was stirred at room temperature for 1.5 hr. The mixture was
concentrated in vacuo, made basic by sat. NaHCO.sub.3, and
extracted with CHCl.sub.3. The extract was washed with brine, dried
over K.sub.2CO.sub.3, and evaporated to give methyl
4-[(4S)-methoxycarbonyl phenoxy)-(2S)-pyrrolidinylmethoxy]benzoate
(1.50 g, q.y.) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.87-1.92 (m, 1H), 2.41-2.48 (m, 1H), 3.18-3.23 (m, 1H),
3.34-3.37 (m, 1H), 3.60-3.66 (m, 1H), 3.88 (s, 3H), 3.89 (s, 3H),
4.04-4.13 (m, 2H), 4.94-5.00 (m, 1H), 6.87-6.93 (m, 4H), 7.96-8.00
(m, 4H); MS (ESI) m/z 386 (M.sup.++1).
[0793] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (400 mg,
1.27 mmol), methyl
4-[(4S)-4-methoxycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate
(491 mg, 1.27 mmol), EDC.HCl (293 mg, 1.53 mmol), HOBt (207 mg,
1.53 mmol), and Et.sub.3N (215 .mu.l, 1.54 mmol) in THF (10 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (60:1 to 50:1, v/v) as eluent to give methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-4-methoxy-
carbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate (532 mg, 61%) as
a white foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.26-2.49 (series of
s and m, total 5H), 3.56-3.93 (series of s and m, total 13H),
4.07-4.59 (series of m, total 3H), 5.01 (m, 1H), 6.69-6.94 (m, 7H),
7.09-7.13 (m, 1H), 7.20-7.31 (m, 3H), 7.52-7.57 (m, 1H), 7.92-8.00
(m, 4H), 8.06-8.09 (m, 1H); MS (ESI) m/z 682 (M.sup.++1).
[0794] To a stirred solution of methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(4S)-(4-methox-
ycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate (532 mg, 0.78
mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the reaction
mixture was heated under reflux for 5 hr. After cooled to room
temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected. The crude solid was
recrystallized from MeOH--CHCl.sub.3-Et.sub.2O to give 116 (125 mg,
25%) as a pale yellow crystalline powder. MW 653.68 mp
154-159.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) 2.24 (s, 3H),
2.38-2.49 (m, 2H), 3.63 (s, 2H), 3.67-3.88 (series of s and m,
total 4H), 4.01-4.06 and 4.15-4.19 (each m, total 2H), 4.27-4.31
and 4.38-4.42 (each m, total 2H), 5.18-5.25 (m, 1H), 6.72-6.77 (m,
1H), 6.85-7.16 (series of m, total 8H), 7.78-7.89 (m, 5H),
7.99-8.02 (m, 1H), 8.46 (s, 1H), 8.57 (s, 1H), 12.65 (br s, 2H); MS
(ESI) m/z 654 (M.sup.++1); Anal. Calcd for
C.sub.36H.sub.35N.sub.3O.sub.9.1/2H.sub.2O: C, 65.25; H, 5.48; N,
6.34. Found: C, 65.29; H, 5.54; N, 6.20.
Example 109
4-[(4S)-(4-carboxyphenoxy)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxypheny-
lacetyl]-(2S)-pyrrolidinylmethoxy]benzoic acid
[0795] ##STR1738##
[0796] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (420 mg,
1.25 mmol), methyl
4-[(4S)-4-methoxycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate
(483 mg, 1.25 mmol), EDC.HCl (288 mg, 1.50 mmol), HOBt (203 mg,
1.50 mmol), and Et.sub.3N (210 .mu.l, 1.51 mmol) in THF (10 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (50:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-(4-methox-
ycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate (488 mg, 55%)
as a white foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.28-2.51 (m,
2H), 3.62-3.94 (series of s and m, total 13H), 4.07-4.62 (series of
m, total 3H), 4.99-5.03 (m, 1H), 6.78-6.99 (m, 7H), 7.23-7.34 (m,
2H), 7.42-7.52 (m, 2H), 7.92-8.01 (m, 5H), 8.17-8.20 (m, 1H); MS
(ESI) m/z 702 (M.sup.++1).
[0797] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-(4-methox-
ycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate (488 mg, 0.70
mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the reaction
mixture was heated under reflux for 3 hr. After cooled to room
temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected. The crude solid was
recrystallized from MeOH--CHCl.sub.3-Et.sub.2O to give 117 (137 mg,
29%) as a white crystalline powder. MW 674.10 mp 150-153.degree.
C.; .sup.1H-NMR (DMSO-d.sub.6) 8 and 4.17-4.21 (each m, total 2H),
4.30-4.34 and 4.40-4.45 (each m, total 2H), 5.20-5.27 (m, 1H),
6.77-6.81 (m, 1H), 6.89-6.92 (m, 1H), 7.01-7.08 (m, 5H), 7.27-7.31
(m, 1H), 7.43-7.46 (m, 1H), 7.86-7.92 (m, 4H), 7.97-8.00 (m, 1H),
8.10-8.12 (m, 1H), 8.91 (s, 1H), 8.96 (s, 1H), 12.65 (br s, 2H); MS
(ESI) m/z 674 (M.sup.++1); Anal. Calcd for
C.sub.35H.sub.32ClN.sub.3O.sub.9.1/4H.sub.2O: C, 61.95; H, 4.83; N,
6.19; Cl, 5.22. Found: C, 61.77; H, 4.86; N, 6.13; Cl, 5.49.
Example 110
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-4-carboxyph-
enoxy)-(2S)-pyrrolidinylmethoxy]benzoic acid
[0798] ##STR1739##
[0799] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (464 mg,
1.22 mmol), methyl
4-[(4S)-(4-methoxycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate
(472 mg, 1.22 mmol), EDC.HCl (282 mg, 1.47 mmol), HOBt (200 mg,
1.48 mmol), and Et.sub.3N (205 .mu.l, 1.47 mmol) in THF (10 ml) was
stirred at room temperature overnight The mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (60:1 to 50:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(4S)-(4-methoxycarbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate
(379 mg, 41%) as a white foam. .sup.1H-NMR (CDCl.sub.3) .delta.
2.28-2.51 (m, 2H), 3.59-3.95 (series of s and m, total 13H),
4.07-4.62 (series of m, total 3H), 4.99-5.03 (m, 1H), 6.79-6.95 (m,
7H), 7.27-7.36 (m, 3H), 7.49-7.51 (m, 1H), 7.93-8.01 (m, 5H),
8.11-8.14 (m, 1H); MS (ESI) m/z 747 (M.sup.++1).
[0800] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-(4-methoxy-
carbonylphenoxy)-(2S)-pyrrolidinylmethoxy]benzoate (379 mg, 0.51
mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the reaction
mixture was heated under reflux for 5 hr. After cooled to room
temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected. The crude solid was purified
by preparative TLC to give 118 (51 mg, 14%) as a pale yellow
amorphous solid. MW 718.55 .sup.1H-NMR (DMSO-d.sub.6) a 2.20-2.40
(m, 2H), 3.65-3.89 (series of m, total 6H), 4.02-4.63 (series of m,
total 4H), 5.19-5.26 (m, 1H), 6.74-7.06 (m, 7H), 7.30-7.34 (m, 1H),
7.59-7.61 (m, 1H), 7.83-7.96 (m, 6H), 8.74 (s, 1H), 8.93 (s, 1H);
Anal. Calcd for C.sub.35H.sub.32BrN.sub.3O.sub.9.2H.sub.2O: C,
55.71; H, 4.81; N, 5.57. Found: C, 55.92; H, 4.80; N, 5.30.
Example 111
4-[(4S)-(4-carboxyphenoxy)-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(-
2S)-pyrrolidinyl methoxybenzoic acid
[0801] ##STR1740##
[0802] A mixture of 4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(328 mg, 1.15 mmol), methyl
4-[(4S)-4-methoxycarbonylphenoxy)-(2S)-pyrrolidinylmethoxybenzoate
(444 mg, 1.15 mmol), EDC.HCl (265 mg, 1.38 mmol), HOBt (187 mg,
1.38 mmol), and Et.sub.3N (195 .mu.l, 1.40 mmol) in THF (10 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (60:1 to 50:1, v/v) as eluent to give methyl
4-[(4S)-(4-methoxycarbonylphenoxy)-1-[4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]-(2S)-pyrrolidinylmethoxy]benzoate (332
mg, 44%) as a white foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.13 (s,
3H), 2.24-2.48 (m, 2H), 3.52-3.90 (series of s and m, total 10H),
4.05-4.58 (series of m, total 3H), 5.01 (m, 1H), 6.78-6.90 (m, 4H),
6.98-7.20 (m, 8H), 7.51-7.56 (m, 2H), 7.90-8.00 (m, 4H); MS (ESI)
m/z 652 (M.sup.++1).
[0803] To a stirred solution of methyl
4-[(4S)-(4-methoxycarbonylphenoxy)-1-[4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]-(2S)-pyrrolidinylmethoxy]benzoate (332
mg, 0.51 mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the
reaction mixture was heated under reflux for 3 hr. After cooled to
room temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected. The crude solid was
recrystallized from MeOH--CHCl.sub.3-Et.sub.2O to give 119 (118 mg,
37%) as a white crystalline powder. MW 623.65 mp 157-160.degree.
C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.20-2.25 (series of s and
m, total 4H), 2.39-2.47 (m, 1H), 3.64 (s, 2H), 3.68-3.89 (m, 1H),
4.02-4.08 and 4.16-4.20 (each m, total 2H), 4.29-4.33 and 4.39-4.43
(each m, total 2H), 5.20-5.26 (m, 1H), 6.92-6.96 (m, 1H), 7.02-7.08
(m, 4H), 7.12-7.18 (m, 4H), 7.39-7.41 (m, 2H), 7.84-7.92 (m, 6H),
9.01 (s, 1H), 12.65 (br s, 2H); MS (ESI) m/z 624 (M.sup.++1); Anal.
Calcd for C.sub.35H.sub.33N.sub.3O.sub.8.1H.sub.2O: C, 65.51; H,
5.50; N, 6.55. Found: C, 65.48; H, 5.36; N, 6.52.
Example 112
4-[4-(2,4-difluorophenoxy)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]pheny-
lacetyl]-2-pyrrolidinylmethoxybenzoic acid
[0804] ##STR1741##
[0805] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-4-hydroxyproline (4.0 g, 16.3 mmol),
Ph.sub.3P (5.14 g, 19.6 mmol), and 2,4-difluorophenol (2.55 g, 19.6
mmol) in THF (50 ml) was added DIAD (3.9 ml, 19.6 mmol), and the
mixture was heated at reflux for 3 h. After cooling to room
temperature, the mixture was concentrated in vacuo and the residue
was chromatographed on silica gel with CHCl.sub.3-EtOAc (4:1) to
give methyl
1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)pyrrolidine-2-carboxylate
(5.82 g, quant) as yellow oil.
[0806] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)pyrrolidine-2-carboxylate
(5.82 g, 16.3 mmol) in THF (130 ml) was added 0.25 N NaOH (130 ml,
32.6 mmol). The resulting mixture was stirred overnight. The
mixture was poured into 1 N HCl (100 ml) and extracted with
CHCl.sub.3 (2.times.200 ml). The extracts were dried over
MgSO.sub.4 and evaporated. The residue was chromatographed on
silica gel with CHCl.sub.3-EtOAc (4:1) as eluent to give
1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)pyrrolidine-2-carb-
oxylic acid (2.55 g, 46%) as a colorless foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.42-1.47 (m, 9H), 2.29-2.74 (series of m,
2H), 3.66-3.71 (m, 2H), 4.46-4.51 (m, 1H), 4.83 (m, 1H), 6.73-6.95
(m, 3H).
[0807] To a stirred solution of
1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)pyrrolidine-2-carboxylic
acid (2.55 g, 7.43 mmol) in THF (50 ml) was added BH.sub.3-DMS (452
ul, 7.43 mmol). The mixture was heated at reflux overnight. After
cooling to room temperature, the mixture was concentrated in vacuo
and quenched by the addition of H.sub.2O (100 ml). The mixture was
extracted with CHCl.sub.3 (2.times.200 ml), dried over MgSO.sub.4,
and evaporated. The residue was chromatographed on silica gel with
CHCl.sub.3-EtOAc (4:1) as eluent to give
1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)-2-pyrrolidinylmethanol
(1.76 g, 72%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.45 (s, 9H), 2.28-2.36 (m, 2H), 3.58-4.99 (series of m, 8H),
6.74-6.90 (m, 3H).
[0808] To a stirred solution of
1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)-2-pyrrolidinylmethanol
(500 mg, 1.52 mmol), methyl 4-hydroxybenzoate (277 mg, 1.82 mmol),
and Ph.sub.3P (477 mg, 1.82 mmol) in THF (10 ml) was added DIAD
(358 ul, 1.82 mmol), and the mixture was heated at reflux for 5 h.
After cooling to room temperature, the mixture was concentrated in
vacuo and the residue was chromatographed on silica gel with
CHCl.sub.3-EtOAc (20:1) as eluent to give methyl
4-[1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)-2-pyrrolidinylmethoxy]-
benzoate (529 mg, 75%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.46 (s, 9H), 2.20-2.47 (m, 2H), 3.64 (m, 2H), 3.86 (s,
3H), 4.07-4.43 (m, 3H), 4.86 (m, 1H), 6.74-6.87 (m, 3H), 6.94 (d,
2H, J=8.5 Hz), 7.95 (d, 2H, J=8.5 Hz).
[0809] To a stirred solution of methyl
4-(1-(tert-butoxycarbonyl)-4-(2,4-difluorophenoxy)-2-pyrrolidinyl
methoxy]benzoate (529 mg, 1.15 mmol) in CH.sub.2Cl.sub.2 (5 ml) was
added TFA (5 ml). The mixture was stirred overnight. The mixture
was concentrated in vacuo and the residue was made basic by the
addition of sat. NaHCO.sub.3. The mixture was extracted with
CHCl.sub.3 (2.times.100 ml). The extracts were dried over
K.sub.2CO.sub.3 and evaporated to give methyl
4-[4-(2,4-difluorophenoxy)-2-pyrrolidinyl methoxy]benzoate (385 mg,
92%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.89-1.95
(m, 1H), 2.28-2.35 (m, 1H), 3.09 (dd, J=12.5, 4.9 Hz, 1H), 3.33 (d,
J=12.5 Hz, 1H), 3.60 (m, 1H), 3.86 (s, 3H), 4.10 (d, J=5.6 Hz, 2H),
4.84 (m, 1H), 6.73-6.89 (m, 3H), 6.91 (d, J=8.5 Hz, 2H), 7.96 (d,
J=8.5 Hz, 2H).
[0810] A mixture of methyl
4-[4-(2,4-difluorophenoxy)-2-pyrrolidinylmethoxy]benzoate (380 mg,
1.05 mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (329 mg, 1.05 mmol), EDC.HCl (302 mg, 1.58 mmol), and
catalytic amount of HOBt and DMAP in DMF (10 ml) was stirred for 3
days. The mixture was diluted with EtOAc (200 ml) and washed with
brine (2.times.200 ml). After removal of the solvent, residue was
chromatographed on silica gel with CHCl.sub.3-EtOAc (4:1) to
CHCl.sub.3-MeOH (10:1) as eluent to give methyl
4-[4-(2,4-difluorophenoxy)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phen-
ylacetyl]-2-pyrrolidinylmethoxybenzoate (693 mg, quant).
.sup.1H-NMR (CDCl.sub.3) .delta.2.16-2.53 (m, 5H), 3.61-4.93
(series of m, 14H), 6.48-8.12 (series of m, 16H).
[0811] To a stirred solution of methyl
4-[4-(2,4-difluorophenoxy)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phen-
ylacetyl]-2-pyrrolidinylmethoxybenzoate (693 mg, 1.05 mmol) in THF
(8 ml) was added 0.25 N NaOH (8.4 ml, 2.10 mmol). The mixture was
stirred overnight. The mixture was poured into 1 N HCl (200 ml) and
the resulting precipitate was collected with suction. The solid was
chromatographed on silica gel with CHCl.sub.3-MeOH (50:1 to 10:1)
as eluent to give 120 (323 mg, 48%) as a colorless amorphous solid.
MW 645.65 .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25 (s, 3H), 2.35 (m,
2H), 3.33-5.18 (series of m, 1H), 6.75 (dd, 1H, J=8.3, 1.7 Hz),
6.87-7.30 (series of m, 8H), 7.79 (d, 1H, J=8.3 Hz), 7.85-7.90 (m,
3H), 8.01 (d, 1H, J=8.3 Hz), 8.49 (s, 1H), 8.57 (s, 1H); MS (FAB)
m/z, 646 (M.sup.++1).
Example 113
4-[1-[4-[N'-2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-(6-quinolyloxy-
-2S-pyrrolidinyl]methoxybenzoic acid
[0812] ##STR1742##
[0813] To a stirred solution of methyl
(trans-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinyl)methoxy
benzoate (1.0 g, 3.0 mmol), 6-hydroxyquinoline (435 mg, 3.0 mmol),
and Ph.sub.3P (943 mg, 3.6 mmol) in THF (10 ml) was added DIAD (727
mg, 3.6 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 18 hr. The mixture was concentrated in vacuo.
The residue was purified by column chromatography on silica gel
with n-hexane-EtOAc (1:2, v/v). To a stirred solution of the
product in CH.sub.2Cl.sub.2 (6.0 ml) was added TFA (6.0 ml) at
0.degree. C. The reaction mixture was stirred at room temperature
for 0.5 hr. The mixture was concentrated in vacuo. Sat. NaHCO.sub.3
was added to the residue, and extracted with CH.sub.2Cl.sub.2. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with MeOH--CH.sub.2Cl.sub.2 (1% to
10%, v/v) as eluent to give methyl
4-[(4S)-[(6-quinolyloxy-(2S)-pyrrolidinyl)]methoxybenzoate (900 mg,
82%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.92-2.10 (m, 1H), 2.45-2.55 (m, 1H), 3.20-3.30 (m, 1H), 3.38-3.50
(m, 1H), 3.60-3.70 (m, 1H), 3.88 (s, 3H), 4.05-4.18 (m, 2H), 5.03
(m, 1H), 6.91 (d, J=8.5 Hz, 1H), 7.02 (d, J=2.7 Hz, 1H), 7.35-7.38
(m, 2H), 7.96 (d, J=8.5 Hz, 1H), 8.00-8.05 (m, 2H), 8.76 (d, J=3.2
Hz, 1H).
[0814] To a stirred solution of methyl
4-(4S-(6-quinolyloxy-2S-pyrrolidinyl)methoxybenzoate (300 mg, 0.79
mmol), 4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid
(264 mg, 0.79 mmol), HOBt (107 mg, 0.79 mmol), and triethylamine
(330 ml, 2.37 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added
EDC.HCl (228 mg, 1.2 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 16 hr, and concentrated in vacuo.
Water was added to the residue, and extracted with EtOAc. The
extract was washed with sat. NaHCO.sub.3 then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with EtOAc to
EtOH-EtOAc (10%, v/v) as eluent to give methyl
4-[1-[4-N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-(6-quinolylox-
y-(2S)-pyrrolidinyl]methoxybenzoate (520 mg, 95%) as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.30-2.60 (m, 3H), 3.64 (s,
2H), 3.73 (s, 3H), 3.80-3.95 (m, 1H), 3.87 (s, 3H), 4.15-4.30 (m,
1H), 4.50-4.70 (m, 2H), 5.11 (br s, 1H), 6.81-7.01 (m, 6H),
7.26-7.39 (m, 6H), 7.93-8.03 (m, 5H), 8.19 (d, J=8.3 Hz, 1H), 8.80
(s, 1H).
[0815] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-4-(6-quinolyloxy-2S-pyrrolidinyl]methoxybenzoate (520 mg,
0.75 mmol) in THF (10.0 ml) and MeOH (5.0 ml) was added 1N NaOH
(1.5 ml, 1.5 mmol). The mixture was stirred at 60.degree. C. for 18
hr. The mixture was concentrated in vacuo, water was added thereto,
and neutralized with 1N HCl. The resulting solid was collected,
washed with water, and dried in vacuo to give 121 (450 mg, 88%) as
a white crystalline solid. 681.13 mp 129-133.degree. C.; IR (KBr)
3332, 1704, 1604, 1531, 1419, 1222, 1166 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.25-2.55 (m, 2H), 3.67 (s, 2H), 3.82 (s,
3H), 3.81-3.92 (m, 1H), 4.02-4.15 (m, 2H), 4.40-4.50 (m, 2H),
5.25-5.40 (m, 1H), 5.33-7.07 (m, 5H), 7.26-7.49 (m, 5H), 7.83-8.23
(m, 6H), 8.73-8.74 (m, 1H), 8.90 (s, 1H), 8.94 (s, 1H); MS (FAB)
m/z 681 (M.sup.++1); Anal. calcd for
C.sub.37H.sub.33N.sub.4O.sub.7Cl.0.5H.sub.2O: C, 64.39; H, 4.97; N,
8.12. Found: C, 64.22; H, 4.90; N, 7.96.
Example 114
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-(6-quinolyl-
oxy-(2S)-pyrrolidinyl]methoxybenzoic acid
[0816] ##STR1743##
[0817] To a stirred solution of methyl
4-(4S-(6-quinolyloxy-2S-pyrrolidinyl)methoxybenzoate (300 mg, 0.79
mmol), 4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (299
mg, 0.79 mmol), HOBt (107 mg, 0.79 mmol), and triethylamine (330
ml, 2.37 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added
EDC.HCl (228 mg, 1.2 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 16 hr, and concentrated in vacuo.
Water was added to the residue, and extracted with EtOAc. The
extract was washed with sat. NaHCO.sub.3 then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with EtOAc to
EtOH-EtOAc (100%, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4S)-(6-quinoly-
loxy-(2S)-pyrrolidinyl]methoxybenzoate (530 mg, 91%) as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.30-2.62 (m, 3H), 3.65 (s,
2H), 3.75 (s, 3H), 3.80-3.95 (m, 1H), 3.93 (s, 3H), 4.10-4.30 (m,
1H), 4.50-4.70 (m, 2H), 5.11 (br s, 1H), 6.82-6.98 (m, 6H),
7.15-7.39 (m, 5H), 7.52 (d, J=8.0 Hz, 1H), 7.93-8.03 (m, 5H), 8.14
(d, J=8.3 Hz, 1H), 8.80 (s, 1H).
[0818] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-4-(6-quinolyloxy-2S-pyrrolidinyl]methoxybenzoate (530 mg,
0.72 mmol) in THF (10.0 ml) and MeOH (5.0 ml) was added 1N NaOH
(1.4 ml, 1.4 mmol). The mixture was stirred at 70.degree. C. for 24
hr. The mixture was concentrated in vacuo, water was added thereto,
and neutralized with 1N HCl. The resulting solid was collected,
washed with water, and dried in vacuo to give 122 (460 mg, 88%) as
a white crystalline solid. MW 725.59 mp 149-153.degree. C.; IR
(KBr) 3332, 1704, 1604, 1527, 1222, 1164 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.28-2.58 (m, 2H), 3.67 (s, 2H), 3.82 (s,
3H), 3.85-3.90 (m, 1H), 4.05-4.15 (m, 2H), 4.40-4.50 (m, 2H),
5.20-5.32 (m, 1H), 6.77-7.07 (m, 5H), 7.31-7.61 (m, 5H), 7.83-7.97
(m, 5H), 8.21-8.22 (m, 1H), 8.73-8.74 (m, 2H), 8.92 (s, 1H); MS
(FAB) m/z 725 (M.sup.+), 727 (M.sup.++2); Anal. calcd for
C.sub.37H.sub.33N.sub.4O.sub.6Br.0.5H.sub.2O: C, 60.50; H, 4.67; N,
7.63; Br, 10.88. Found: C, 60.51; H, 4.60; N, 7.52; Br, 11.06.
Example 115
4-[(2S,4S)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(2-na-
phthyloxy)-2-pyrrolidinyl]methoxybenzoic acid
[0819] ##STR1744##
[0820] To a stirred mixture of methyl
(2S,4R)-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinylcarboxylate
(4.22 g, 17.2 mmol), 2-naphthol (2.73 g, 18.9 mmol) and PPh.sub.3
(4.96 g, 18.9 mmol) in THF (80 ml) was added DIAD (3.72 ml, 18.9
mmol) at room temperature under an atmosphere of nitrogen. After
stirring over night, the mixture was concentrated in vacuo. The
residue was chromatographed on silica gel [600 g, CHCl.sub.3/EtOAc
(10/1)], to give methyl
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylcarboxylate
(5.37 g), which was used without further purification.
[0821] To a stirred solution of methyl
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinyl
carboxylate (5.37 g) in THF (116 ml) was added 0.25 N NaOH (116 ml,
29.0 mmol) at room temperature. The resulting mixture was stirred
over night. After removal of the solvent, the mixture was acidified
by the addition of 1 N HCl and extracted with CHCl.sub.3. The
combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was recrystallized
from n-hexane-CHCl.sub.3, to give
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylcarboxylic
acid [4.44 g, 85% (2 steps)] as a white powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.37 and 1.41 (s, 9H, amide isomers), 2.26
(d, J=13.9 Hz, 1H), 2.65 (m, 1H), 3.47 (d, J=11.5 Hz, 1H), 3.81 (m,
1H), 4.30 (m, 1H), 5.14 (m, 1H), 7.02-7.86 (m, 7H).
[0822] To a stirred solution of
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylcarboxylic
acid (1.12 g, 3.13 mmol) in THF (30 ml) was added BH.sub.3-DMS
(0.63 ml, 6.3 mmol) at 0.degree. C. The mixture was raised to room
temperature immediately and then heated at 50.degree. C. for 1.5 h.
After cooling to room temperature, the mixture was quenched by the
addition of water at 0.degree. C. and extracted with EtOAc. The
combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/MeOH (50/1)], to give
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylmethanol
(1.10 g, 100%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.48 (s, 9H), 2.45 (m, 1H), 3.58-4.80 (m, 4H), 5.01 (br,
1H), 7.04-7.99 (m, 7H).
[0823] To a stirred mixture of
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinyl
methanol (640 mg, 1.86 mmol), methyl 4-hydroxybenzoate (283 mg,
1.86 mmol) and PPh.sub.3 (488 mg, 1.86 mmol) in THF (18 ml) was
added DIAD (0.37 ml, 1.86 mmol) at room temperature under an
atmosphere of nitrogen. The mixture was stirred over night. After
removal of the solvent, the resulting residue was chromatographed
on silica gel [100 g, n-hexane/EtOAc (2/1)], to give methyl
4-[(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxy-
benzoate (830 mg, 93%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.50 (d, J=8.3 Hz, 9H), 2.34 (m, 1H), 2.53 (d, J=14.2 Hz,
1H), 3.72-3.85 (m, 1H), 3.86 and 3.87 (s, 3H, amide isomers), 4.17
(m, 1H), 4.26-4.52 (m, 2H), 5.06 (br, 1H), 6.87 (d, J=8.8 Hz, 1H),
6.94 (d, J=8.8 Hz, 2H), 7.04 (br, 2H), 7.33 (t, J=7.3 Hz, 1H), 7.42
(t, J=7.3 Hz, 1H), 7.64-8.02 (m, 5H).
[0824] To a stirred solution of methyl
4-[(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxy-
benzoate (870 mg, 1.74 mmol) in CH.sub.2Cl.sub.2 (24 ml) was added
TFA (6 ml) at room temperature. The mixture was stirred over night,
which was concentrated in vacuo. The residue was diluted with
CH.sub.2Cl.sub.2 and made basic by the addition of 1 N NaOH, which
was extracted with CH.sub.2Cl.sub.2. The organic layer was washed
with brine, dried over NaSO.sub.4 and concentrated. The residue was
chromatographed on silica gel [100 g, n-hexane/EtOAc (2/1)], to
give methyl
4-[(2S,4S)-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxybenzoate (750
mg, 100%) as a black oil .sup.1H-NMR (CDCl.sub.3) .delta. 1.99 (dd,
J=14.2, 5.6 Hz, 1H), 2.48 (m, 1H), 3.22 (dd, J=12.2, 4.6 Hz, 1H),
3.43 (d, J=12.5 Hz, 1H), 3.67 (m, 1H), 3.86 and 3.87 (s, 3H, amide
isomers), 4.11 (m, 2H), 5.04 (m, 1H), 6.83 (d, J=8.5 Hz, 1H), 6.89
(d, J=8.8 Hz, 2H), 7.07 (d, J=2.0 Hz, 1H), 7.12 (d, J=9.0 Hz, 1H),
7.33 (dt, J=8.1, 1.2 Hz, 1H), 7.44 (dt, J=6.8, 1.2 Hz, 1H), 7.70
(d, J=8.1 Hz, 1H), 7.75 (dd, J=9.0, 5.1 Hz, 2H), 7.90 (d, J=8.5 Hz,
1H), 7.96 (dd, J=6.8, 2.0 Hz, 2H).
[0825] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (333 mg,
0.106 mmol), methyl
4-[(2S,4S)-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxybenzoate (400
mg, 1.06 mmol), EDC.HCl (305 mg, 1.59 mmol) and DMAP (194 mg, 1.59
mmol) in DMF (10 ml) was stirred at room temperature for 3 days.
The mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [100 g, n-hexane/EtOAc
(1/1) CHCl.sub.3/MeOH (50/1)], to give methyl
4-[(2S,4S)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(2-n-
aphthyloxy)-2-pyrrolidinyl]methoxybenzoate (520 mg, 73%) as a pale
brown amorphous. .sup.1H-NMR (CDCl.sub.3) .delta. 2.28 (s, 3H),
2.29 (m, 1H), 2.55 (d, J=14.2 Hz, 1H), 3.60 (d, J=3.4 Hz, 2H), 3.66
(d, J=3.7 Hz, 3H), 3.68-4.00 (m, 5H), 4.05-4.67 (m, 3H), 5.09 (br,
1H), 6.61 (s, 1H), 6.77 (m, 2H), 6.87 (d, J=8.8 Hz, 1H), 6.94-7.54
(m, 8H), 7.68-8.09 (m, 8H); MS (ESI) m/z 674 (M.sup.++1).
[0826] To a solution of methyl
4-[(2S,4S)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido)phenylacetyl]-4-(2-n-
aphthyloxy)-2-pyrrolidinyl]methoxybenzoate (415 mg, 0.616 mmol) in
THF (4.9 ml), 0.25 N NaOH (4.9 ml) was added. After stirring at
room temperature for 3 days, the mixture was acidified with 1 N HCl
and extracted with CHCl.sub.3-MeOH (10/1). The combined extracts
were dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The
residue was purified on TLC [CHCl.sub.3/MeOH (10/1)], to give 123
(180 mg, 44%) as a colorless amorphous. MW 659.73 IR (KBr) 3354,
2937, 1685, 1601, 1533, 1255 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.24 (s, 3H), 2.25-2.43 (m, 2H), 3.65 (s, 2H), 3.81 (s,
3H), 3.83 (m, 1H), 4.05-4.70 (m, 4H), 5.21-5.33 (br, 1H), 6.76 (d,
J=7.3 Hz, 1H), 6.86-7.35 (m, 9H), 7.44 (t, J=7.3 Hz, 1H), 7.76-7.89
(m, 6H), 8.01 (d, J=8.3 Hz, 1H), 8.48 (s, 1H), 8.56 (s, 1H); MS
(FAB) m/z 660 (M.sup.++1).
Example 116
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl)-4-(2-na-
phthyloxy)-2-pyrrolidinyl]methoxybenzoic acid
[0827] ##STR1745##
[0828] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (310 mg,
0.93 mmol), methyl
4-[(2S,4S)-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxybenzoate (350
mg, 0.93 mmol), EDC.HCl (267 mg, 1.40 mmol) and DMAP (171 mg, 1.40
mmol) in DMF (10 ml) was stirred at room temperature for 3 days.
The mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [100 g, n-hexane/EtOAc
(1/1) CHCl.sub.3/MeOH (50/1)], to give methyl
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-(2-n-
aphthyloxy)-2-pyrrolidinyl]methoxybenzoate (450 mg, 68%) as a pale
brown amorphous. .sup.1H-NMR (CDCl.sub.3) .delta. 2.32 (m, 1H),
2.58 (d, J=14.5 Hz, 1H), 3.63 (d, J=2.7 Hz, 1H), 3.70 (s, 3H), 3.86
(s, 3H), 3.84-3.95 (m, 2H), 4.15-4.64 (m, 4H), 5.11 (br, 1H),
6.79-7.06 (m, 7H), 7.21-7.46 (m, 7H), 7.66-7.77 (m, 3H), 7.92 (d,
J=8.8 Hz, 1H), 7.97 (m, 1H), 8.17 (d, J=8.4 Hz, 1H); MS (ESI) m/z
694 (M.sup.++1), 696 (M.sup.++3).
[0829] To a solution of methyl
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxybenzoate (381 mg,
0.535 mmol) in THF (4.3 ml), 0.25 N NaOH (4.3 ml) was added. After
stirring at room temperature for 3 days, the mixture was acidified
with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1). The
combined extracts were dried over Na.sub.2SO.sub.4 and concentrated
in vacuo. The residue was purified on TLC [CHCl.sub.3/MeOH (10/1)]
to give 124 (140 mg, 39%) as a colorless amorphous. MW 680.15 IR
(KBr) 3323, 2935, 1704, 1601, 1529, 1529, 1508 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.27-2.49 (m, 2H), 3.65 (s, 2H),
3.81 (s, 3H), 3.83 (m, 1H), 4.05-4.71 (m, 4H), 5.30 (br, 1H), 6.77
(d, J=7.8 Hz, 1H), 6.87-7.16 (m, 4H), 7.14 (dd, J=8.8, 2.2 Hz, 1H),
7.27 (t, J=7.3 Hz, 1H), 7.29-7.46 (m, 4H), 7.76-7.86 (m, 5H), 7.96
(d, J=8.3 Hz, 1H), 8.08 (dd, J=8.3, 1.2 Hz, 1H), 8.90 (s, 1H), 8.93
(s, 1H); MS (FAB) m/z 680 (M.sup.++1), 682 (M.sup.++3); Anal. Calcd
for C.sub.38H.sub.34ClN.sub.3O.sub.7.1H.sub.2O: C, 65.37; H, 5.20;
N, 6.02. Found: C, 65.43; H, 5.11; N, 5.93.
Example 117
2-[(2S,4S)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(2-na-
phthyloxy)-2-pyrrolidinyl]methoxy-5-pyridinecarboxylic acid
[0830] ##STR1746##
[0831] To a stirred mixture of methyl
(2S,4R)-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinylcarboxylate
(4.22 g, 17.2 mmol), 2-naphthol (2.73 g, 18.9 mmol) and PPh.sub.3
(4.96 g, 18.9 mmol) in THF (80 ml) was added DIAD (3.72 ml, 18.9
mmol) at room temperature under an atmosphere of nitrogen. After
stirring over night, the mixture was concentrated in vacuo. The
residue was chromatographed on silica gel [600 g, CHCl.sub.3/EtOAc
(10/1)], to give methyl
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylcarboxylate
(5.37 g), which was used to the next reaction without further
purification.
[0832] To a stirred solution of methyl
(2S,4S)-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinyl
carboxylate (5.37 g) in THF (116 ml) was added 0.25 N NaOH (116 ml,
29.0 mmol) at room temperature. The resulting mixture was stirred
over night. After removal of the solvent, the mixture was acidified
by the addition of 1 N HCl and extracted with CHCl.sub.3. The
combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was recrystallized
with n-hexane-CHCl.sub.3, to give
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylcarboxylic
acid [4.44 g, 85% (2 steps)] as a white powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.37 and 1.41 (s, 9H, amide isomers), 2.26
(d, J=13.9 Hz, 1H), 2.65 (m, 1H), 3.47 (d, J=11.5 Hz, 1H), 3.81 (m,
1H), 4.30 (m, 1H), 5.14 (m, 1H), 7.02-7.86 (m, 7H).
[0833] To a stirred solution of
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylcarboxylic
acid (1.12 g, 3.13 mmol) in THF (30 ml) was added BH.sub.3-DMS
(0.63 ml, 6.3 mmol) at 0.degree. C. The mixture was raised to room
temperature immediately and then heated at 50.degree. C. for 1.5 h.
After cooling to room temperature, the mixture was quenched by the
addition of water at 0.degree. C. and extracted with EtOAc. The
combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/MeOH (50/1)], to give
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylmethanol
(1.10 g, 100%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.48 (s, 9H), 2.45 (m, 1H), 3.58-4.80 (m, 4H), 5.01 (br,
1H), 7.04-7.99 (m, 7H).
[0834] To a stirred mixture of
(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinylmethanol
(484 mg, 1.41 mmol), methyl 2-hydroxy-5-pyridinecarboxylate (216
mg, 1.41 mmol) and PPh.sub.3 (370 mg, 1.41 mmol) in THF (15 ml) was
added DIAD (0.28 ml, 1.41 mmol) at room temperature under an
atmosphere of nitrogen. The mixture was stirred over night. After
removal of the solvent, the resulting residue was chromatographed
on silica gel [50 g, n-hexane/EtOAc (2/1)], to give
methyl-2-[(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrrolidinyl]-
methoxypyridine-5-carboxylate (170 mg, 25%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.47 (s, 9H), 2.37 (m, 1H), 2.46
(d, J=14.2 Hz, 1H), 3.71-4.00 (m, 2H), 3.89 (s, 3H), 4.30-4.56 (m,
2H), 4.74 (dd, J=9.8, 4.6 Hz, 1H), 5.06 (br, 1H), 6.70 (d, J=8.8
Hz, 2H), 7.05-7.09 (m, 2H), 7.33 (t, J=6.9 Hz, 1H), 7.42 (t, J=6.9
Hz, 1H), 7.67-7.75 (m, 3H), 8.09 (d, J=8.8 Hz, 1H), 8.77 (d, J=2.2
Hz, 1H).
[0835] To a stirred solution of
5-carboxymethyl-2-[(2S,4S)-1-tert-butoxycarbonyl-4-(2-naphthyloxy)-2-pyrr-
olidinyl]methoxypyridine (170 mg, 0.36 mmol) in CH.sub.2Cl.sub.2 (5
ml) was added TFA (2 ml) at room temperature. After 2 h stirring,
the mixture was concentrated in vacuo, which was diluted with
CH.sub.2Cl.sub.2 and basified by the addition of 1 N NaOH, The
combined reaction mixture was extracted with CH.sub.2Cl.sub.2. The
organic layer was washed with brine, which were dried over
NaSO.sub.4 and concentrated. The residue was purified on TLC
[CHCl.sub.3/MeOH (10/1)], to give methyl
2-[(2S,4S)-4-(2-naphthyloxy)2-pyrrolidinyl]methoxypyridine-5-carboxylate
(107 mg, 80%) as a colorless oil .sup.1H-NMR (CDCl.sub.3) .delta.
1.95 (m, 1H), 2.27 (br, 1H), 2.46 (m, 1H), 3.19 (dd, J=12.2, 4.9
Hz, 1H), 3.41 (d, J=12.2 Hz, 1H), 3.65 (m, 1H), 3.89 (s, 3H), 4.58
(m, 2H), 5.00 (br, 1H), 6.77 (d, J=8.8 Hz, 1H), 7.06 (br, 1H), 7.11
(dd, J=8.8, 2.7 Hz, 1H), 7.31-7.45 (m, 2H), 7.69-7.76 (m, 3H), 8.13
(dd, J=8.8, 2.4 Hz, 1H), 8.78 (d, J=2.2 Hz, 1H).
[0836] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (89 mg,
0.283 mmol),
methyl-2-[(2S,4S)-4-(2-naphthyloxy)-2-pyrrolidinyl]methoxypyridine-5-carb-
oxylate (107 mg, 0.78 mmol), EDC.HCl (81 mg, 0.425 mmol) and DMAP
(52 mg, 0.425 mmol) in DMF (3 ml) was stirred at room temperature
for 18 h. The mixture was poured into ice water and extracted with
EtOAc. The combined extracts were washed with ice water and brine.
After dried over Na.sub.2SO.sub.4, the extracts were concentrated
in vacuo. The residue was purified on TLC [CHCl.sub.3/MeOH (10/1)],
to give
2-[(2S,4S)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(2-n-
aphthyloxy)-2-pyrrolidinyl]methoxy-5-pyridinecarboxylic acid methyl
ester (193 mg, 100%) as a colorless amorphous. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.27 (d, J=3.2 Hz, 3H), 2.30 (m, 1H), 2.49
(dd, J=14.2, 2.0 Hz, 1H), 3.60 (d, J=3.9 Hz, 1H), 3.67 (d, J=5.9
Hz, 3H), 3.81 (s, 1H), 3.85 (s, 1H), 3.88 and 3.91 (s, 3H, amide
isomers), 3.95 (m, 1H), 4.02-5.09 (m, 4H), 6.67 (d, J=8.8 Hz, 1H),
6.73-7.13 (m, 3H), 7.20-7.45 (m, 7H), 7.53 (t, J=7.8 Hz, 1H),
7.67-7.77 (m, 3H), 8.02-8.84 (m, 3H).
[0837] For HCl salt: a pale brown amorphous. IR (KBr) 3346, 2951,
1720, 1601, 1533, 1281 cm.sup.-1; MS (FAB) m/z 675 (M.sup.++1).
[0838] To a solution of
2-[(2S,4S)-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(2-n-
aphthyloxy)-2-pyrrolidinyl]methoxy-5-pyridinecarboxylic acid methyl
ester (158 mg, 0.23 mmol) in THF (1.8 ml), 0.25 N NaOH (1.8 ml) was
added. After stirring at room temperature for 22 h, the mixture was
neutralized with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1).
The combined extracts were dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was purified on TLC
[CHCl.sub.3/MeOH (5/1)] to give 125 (51 mg, 34%) as a colorless
amorphous solid. MW 660.72 IR (KBr) 3354, 2956, 1601, 1533, 1255,
1022 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.24 (s, 3H),
3.30-5.32 (m, 13H), 6.72-8.82 (m, 19H); MS (FAB) m/z 661
(M.sup.++1); Anal. Calcd for
C.sub.38H.sub.36N.sub.4O.sub.7.0.5EtOH.1H.sub.2O: C, 66.75; H,
5.89; N, 7.98. Found: C, 66.39; H, 5.55; N, 7.66.
Example 118
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-(-
5)-pyrrolidinyl methoxy]benzoic acid
[0839] ##STR1747##
[0840] To a stirred solution of benzyl-(S)-glycidyl ether (5.0 g,
30.5 mmol) in THF (100 ml) was added allylmagnesium chloride (1.0 M
in Et.sub.2O, 30.5 ml, 30.5 mmol) at -78.degree. C., and the
resulting mixture was gradually warmed up to rt with stirring. The
mixture was poured into water and concentrated in vacuo, then
extracted with CHCl.sub.3. The organic layer was dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was chromatographed on silica gel with hexane-EtOAc (5:1) as eluent
to give 1-benzyloxy-2-(R)-hydroxy-5-hexene (2.18 g, 35%) as a
colorless oil: .sup.1H-NMR (CDCl.sub.3) .delta. 1.52-1.60 (m, 2H),
2.11-2.25 (m, 2H), 2.34 (d, J=3.2 Hz, 1H), 3.35 (dd, J=9.6, 8.0 Hz,
1H), 3.52 (dd, J=9.6, 3.2 Hz, 1H), 3.84-3.86 (m, 1H), 4.57 (s, 2H),
4.96-5.07 (series of m, 2H), 5.78-5.88 (m, 1H), 7.29-7.38 (m, 5H);
MS (ESI) m/z, 224 (M.sup.++NH.sub.4.sup.+).
[0841] To a stirred solution of 1-benzyloxy-2-(R)-hydroxy-5-hexene
(2.18 g, 10.5 mmol), triphenylphosphine (3.32 g, 12.7 mmol) and
phthalimide (1.86 g, 12.7 mmol) was added diisopropyl
azodicarboxylate (2.62 ml, 12.7 mmol) at rt, and the resulting
mixture was stirred for overnight at rt. The mixture was
concentrated in vacuo and extracted with EtOAc. The organic layer
was washed with water, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with hexane-EtOAc (5:1) as eluent to give
1-benzyloxy-2-(S)-phthalimido-5-hexene (2.95 g, 83%) as a colorless
oil: .sup.1H-NMR (CDCl.sub.3) .delta. 1.76-1.84 (m, 2H), 2.06 (dd,
J=14.4, 6.8 Hz), 2H, 2.12-2.22 (m, 1H), 3.69 (dd, J=10.0, 5.6 Hz,
1H), 4.00 (t, J=9.6 Hz, 1H), 4.46 (d, J=12.0 Hz, 1H), 4.53 (d,
J=12.0 Hz, 1H), 4.51-4.58 (m, 1H), 4.91-4.99 (series of m, 2H),
5.72-5.79 (m, 1H) 7.21-7.26 (m, 5H), 7.71-7.83 (series of m, 2H);
MS (ESI) m/z, 336 (M.sup.++H).
[0842] To a stirred solution of
1-benzyloxy-2-(S)-phthalimido-5-hexene (2.95 g, 8.80 mmol) in EtOH
(30 ml) was added hydrazine hydrate (80% in water, 460 ml, 11.4
mmol) at rt, and the resulting mixture was heated under reflux for
7.5 h with stirring. The solution was filtered, and the filtrate
was concentrated in vacuo. The residue was poured into
aq.NaHCO.sub.3 and extracted with CHCl.sub.3. The organic layer was
dried over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo
to give 2-(S)-amino-1-benzyloxy-5-hexene (1.90 mg, quant.) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.37-1.55 (series
of m, 4H), 2.08-2.19 (m, 2H), 2.99-3.03 (m, 1H), 3.25 (dd, J=9.2,
7.6 Hz, 1H), 3.45 (dd, J=9.2, 4.0 Hz, 1H), 4.53 (s, 2H), 4.94-5.06
(series of m, 2H), 5.76-5.85 (m, 1H), 7.27-7.37 (m, 5H); MS (ESI)
m/z, 206 (M.sup.++H), 247 (M.sup.++H+CH.sub.3CN).
[0843] To a stirred solution of 2-(S)-amino-1-benzyloxy-5-hexene
(1.89 g, 9.21 mmol) and triethylamine (1.28 ml, 9.21 mmol) in
CH.sub.2Cl.sub.2 (20 ml) was added benzoyl chloride (1.07 ml, 9.21
mmol) at rt, and the resulting mixture was stirred for 23 h. The
mixture was poured into water and extracted with CH.sub.2Cl.sub.2.
The organic layer was washed with water, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-EtOAc (5:1) as eluent to
give N-[2-(S)-benzyloxy)-5-hexenyl]benzamide (2.67 g, 94%) as a
colorless needles. mp 78-79.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.76-1.82 (m, 2H), 2.11-2.17 (m, 2H), 3.59 (brs, 2H),
4.29-4.35 (m, 1H), 4.54 (dd, J=19.2, 12.0 Hz, 2H), 4.96-5.05
(series of m, 2H), 5.78-5.89 (m, 1H), 6.39 (d, J=8.0 Hz, 1H),
7.27-7.51 (m, 8H), 7.74 (d, J=7.2 Hz, 2H); MS (ESI) m/z, 310
(M.sup.++H).
[0844] To a stirred solution of
N-[2-(S)-benzyloxy)-5-hexenyl]benzamide (2.41 g, 7.79 mmol) in
CH.sub.3CN--H.sub.2O (3:1, 40 ml) was added iodine (2.97 g, 23.4
mmol) in one portion, and the resulting mixture was stirred for 20
h. The mixture was poured into aq.Na.sub.2S.sub.2O.sub.3 and
concentrated in vacuo, then extracted with CHCl.sub.3. The organic
layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
dissolved in CH.sub.2Cl.sub.2 (30 ml) and was added di-tert-butyl
dicarbonate (2.55 g, 11.7 mmol), Et.sub.3N (1.63 ml, 11.7 mmol) and
N,N-dimethyl aminopyridine (180 mg, 1.47 mmol), and the resulting
mixture was stirred overnight at rt. The mixture was poured into
water and extracted with CH.sub.2Cl.sub.2. The organic layer was
washed with water, drying over Na.sub.2SO.sub.4, and concentrated
in vacuo. The residue was chromatographed on silica gel with
hexane-EtOAc (5:1) to give
N-Boc-2-(S)-benzoyloxymethyl-5-(S)-benzyloxy methylpyrrolidine
(1.27 g, 38%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.41 and 1.49 (s, total 9H), 1.85-2.00 (series of m, 4H), 3.33-4.59
(series of m, 8H), 7.26-7.32 (m, 5H), 7.41-7.46 (m, 2H), 7.54-7.57
(m, 1H), 8.02 (d, J=7.6 Hz, 2H); MS (ESI) m/z, 426 (M.sup.++H), 448
(M.sup.++Na.sup.+).
[0845] To a stirred solution of
N-Boc-2-(S)-benzoyloxymethyl-5-(S)-benzyloxymethylpyrrolidine (1.23
g, 2.89 mmol) in MeOH (30 ml) was added NaOH (1.0 M in water, 3.47
ml, 3.47 mmol) at rt, and the resulting mixture was stirred for 4
h. The mixture was neutralized with aq. 1N-HCl and concentrated in
vacuo, then extracted with CHCl.sub.3. The organic layer was washed
with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with hexane-EtOAc (3:1) as eluent to give
N-Boc-5-(S)-hydroxymethyl-2-(S)-benzyloxymethylpyrrolidine (847 mg,
91%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.41 (s,
9H), 1.57 (brs, 1H), 1.95-1.97 (m, 2H), 2.05-2.18 (m, 1H), 3.36 (t,
J=8.4 Hz, 1H), 3.56-3.62 (m, 2H), 3.67-3.72 (m, 2H), 3.95 (brs,
1H), 4.03 (brs, 1H), 4.51 (s, 1H), 7.28-7.37 (m, 5H); MS (FAB) m/z,
322 (M.sup.++H).
[0846] To a stirred solution of
N-Boc-2-(S)-hydroxymethyl-5-(S)-benzyloxymethylpyrrolidine (388 mg,
1.21 mmol), triphenylphosphine (380 mg, 1.45 mmol) and methyl
4-hydroxybenzoate (220 mg, 1.45 mmol) was added diisopropyl
azodicarboxylate (200 ml, 1.45 mmol) at rt, and the resulting
mixture was stirred overnight. The mixture was concentrated in
vacuo and extracted with EtOAc. The organic layer was washed with
water, drying over anhydrous Na.sub.2SO.sub.4, then concentrated in
vacuo. The residue was chromatographed on silica gel with
hexane-EtOAc (3:1) as eluent to give methyl
4-[2-(S)--(N-Boc-5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoat-
e (462 mg, 84%) as a colorless oil: .sup.1H-NMR (CDCl.sub.3)
.delta. 1.40 and 1.48 (s, total 9H), 1.98-2.13 (m, 4H), 3.83 and
3.85 (s, 3H), 3.33-4.25 (series of m, 4H), 4.47-4.59 (m, 2H),
6.91-6.96 (m, 2H), 7.26-7.34 (m, 5H), 7.95-7.98 (m, 2H); MS (FAB)
m/z, 456 (M.sup.++H), 478 (M.sup.++Na.sup.+).
[0847] To a stirred solution of methyl
4-[2-(S)--(N-Boc-5-(S)-benzyloxymethyl)pyrrolidinyl
methoxy]benzoate (446 mg, 0.98 mmol) in CH.sub.2Cl.sub.2 (10 ml)
was added trifluoroaceticacid (10 ml) at rt, and the resulting
mixture was stirred for 1 h. The mixture was concentrated in vacuo
and poured into aq.NaHCO.sub.3, then extracted with CHCl.sub.3. The
organic layer was washed with water, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo to give methyl
4-[2-(S)-5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (363
mg, quant.) as yellowish oil. The product was used for next
reactions without further purification. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.43-1.65 (m, 2H), 1.93-2.07 (m, 3H), 3.36-3.68 (series of
m, 4H), 3.89 (s, 3H), 3.86-3.93 (over lap, 2H), 4.55 (s, 2H), 6.90
(d, J=8.4 Hz, 2H), 7.26-7.37 (m, 5H), 7.97 (d, J=8.4 Hz, 2H); MS
(FAB) m/z, 356 (M.sup.++H).
[0848] To a stirred solution of methyl
4-[2-(S)-(5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (115
mg, 0.32 mmol), 4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(92.0 mg, 0.32 mmol) and N,N-dimethylaminopyridine (52.0 mg, 0.42
mmol) in DMF (10 ml) was added EDC.HCl (81.0 mg, 0.42 mmol) at rt,
and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with CHCl.sub.3. The
organic layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl 4-[5-(R)-benzyl
oxymethyl-1-[4-[N'-(2-methylphenyl)ureido]phenylacetamido]-2-(S)-pyrrolid-
inylmethoxy]benzoate (169 mg, 84%) as a colorless amorphous solid.
.sup.1H-NMR (CDCl.sub.3), mixture of rotamars .delta. 1.92-2.18 (m,
3H), 2.24 and 2.25 (s, total 3H), 2.20-2.31 (overlap, 1H),
3.39-3.70 (series of m, 4H), 3.87 and 3.89 (s, total 3H), 4.17 and
4.18 (s, total 2H), 4.30-4.45 (series of m, 2H), 4.53 (s, 2H),
6.43-7.13 (series of m, 9H), 7.20-7.36 (series of m, 7H), 7.58-7.99
(series of m, 3H); MS (FAB) m/z, 622 (M.sup.++H).
[0849] To a stirred solution of methyl
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2--
(S)-pyrrolidinylmethoxy]benzoate (156 mg, 0.24 mmol) in MeOH-THF
(1:5, 12 ml) was added 1.0M-NaOH (1.2 ml, 1.20 mmol) at rt, and the
resulting mixture was heated at 80.degree. C. with stirring for 7
h. The mixture was poured into 1N-HCl, then extracted with
CHCl.sub.3. The organic layer was washed with brine, drying over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (5:1) as
eluent to give 126 (94.0 mg, 65%) as a colorless amorphous solid.
MW 607.70 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars .delta.
1.85-2.35 (series of m, 4H), 2.43-2.92 (series of m, 5H), 2.28 (s,
3H), 3.55-4.55 (series of m, 10H), 6.85-7.95 (series of m, 17H); MS
(ESI) m/z, 630 (M.sup.++Na.sup.+).
Example 119
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyla-
cetyl]-2-(S)-pyrrolidinylmethoxy]benzoic acid
[0850] ##STR1748##
[0851] To a stirred solution of methyl
4-[2-(S)-5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (117
mg, 0.33 mmol), 4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic
acid (110 mg, 0.33 mmol) and N,N-dimethylaminopyridine (50.0 mg,
0.40 mmol) in DMF (10 ml) was added EDC.HCl (76.0 mg, 0.40 mmol) at
rt, and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with CHCl.sub.3. The
organic layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl-
acetyl]-2-(S)-pyrrolidinyl methoxy]benzoate (189 mg, 85%) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3), mixture of
rotamars .delta. 1.91-2.30 (series of m, 4H), 3.39-3.74 (series of
m, 3H), 3.73 (s, 2H), 4.15-4.02 (m, 2H), 4.32-4.44 (m, 1H), 4.54
(s, 2H), 6.71-7.02 (series of m, 5H), 7.06 (s, 1H), 7.17 (s, 1H),
7.24-7.40 (series of m, 7H), 7.89-8.22 (series of m, 4H); MS (FAB)
m/z, 672 (M.sup.++H).
[0852] To a stirred solution of methyl
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl-
acetyl-2-(S)-pyrrolidinylmethoxy]benzoate (169 mg, 0.25 mmol) in
MeOH-THF (2:5, 7 ml) was added 1.0M-NaOH (750 ml, 0.75 mmol) at rt,
and the resulting mixture was heated at 80.degree. C. with stirring
for 2 h. The mixture was poured into 1N-HCl, then extracted with
CHCl.sub.3. The organic layer was washed with brine, drying over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (15:1) as
eluent to give 127 (114 mg, 69%) as a colorless amorphous solid. MW
658.14 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars .delta.
1.88-2.37 (series of m, 4H), 3.51-4.49 (series of m, 8H), 3.64 and
3.73 (s, total 3H), 4.86 (s, 2H), 6.85-7.95 (series of m, 16H); MS
(ESI) m/z, 658 (M.sup.++H), 680 (M.sup.++Na.sup.+); Anal. Calcd for
C.sub.36H.sub.36ClN.sub.3O.sub.7.H.sub.2O: C, 63.95; H, 5.66; N,
6.21. Found: C, 63.65; H, 5.40; N, 5.95.
Example 120
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylac-
etyl]-2-(S)-pyrrolidinylmethoxy]benzoic acid
[0853] ##STR1749##
[0854] To a stirred solution of methyl
4-[2-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (119 mg, 0.34
mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (127
mg, 0.34 mmol) and N,N-dimethylaminopyridine (50.0 mg, 0.40 mmol)
in DMF (10 ml) was added EDC.HCl (77.0 mg, 0.40 mmol) at rt, and
the resulting mixture was stirred overnight. The reaction mixture
was poured into water and extracted with CHCl.sub.3. The organic
layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyla-
ctyl]-2-(S)-pyrrolidinyl methoxy]benzoate (217 mg, 90%) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3), mixture of
rotamars .delta. 1.92-2.31 (series of m, 4H), 3.39-3.73 (series of
m, 3H), 3.65 (s, 2H), 4.15-4.02 (m, 2H), 4.32-4.44 (m, 1H), 4.54
(s, 2H), 6.71-6.99 (series of m, 5H), 7.04 (s, 1H), 7.11 (s, 1H),
7.22-7.39 (series of m, 7H), 7.51-8.17 (series of m, 4H); MS (FAB)
m/z, 716 (M.sup.+), 718 (M.sup.++2).
[0855] To a stirred solution of methyl
4-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyla-
cetyl]-2-(S)-pyrrolidinylmethoxy]benzoate (178 mg, 0.25 mmol) in
MeOH-THF (2:5, 7 ml) was added 1.0M-NaOH (750 ml, 0.75 mmol) at rt,
and the resulting mixture was heated at 80.degree. C. with stirring
for 1.5 h. The reaction mixture was poured into 1N-HCl, then
extracted with CHCl.sub.3. The organic layer was washed with brine,
drying over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(15:1) as eluent to give 128 (159 mg, 91%) as a colorless amorphous
solid. MW 702.59 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars
.delta. 1.88-2.35 (series of m, 4H), 3.51-4.49 (series of m, 8H),
3.64 and 3.72 (s, total 3H), 4.87 (s, 2H), 6.65-8.05 (series of m,
16H); MS (ESI) m/z, 702 (M.sup.+), 704 (M.sup.++2); Anal. Calcd for
C.sub.36H.sub.36BrN.sub.3O.sub.7.H.sub.2O: C, 60.00; H, 5.32; N,
5.83. Found: C, 59.66; H, 5.04; N, 5.65.
Example 121
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenyla-
cetyl]-2-(S)-pyrrolidinylmethoxy]benzoic acid
[0856] ##STR1750##
[0857] To a stirred solution of
N-Boc-2-(S)-hydroxymethyl-5-(S)-benzyloxymethylpyrrolidine (415 mg,
1.29 mmol), triphenylphosphine (410 mg, 1.55 mmol) and methyl
3-hydroxybenzoate (240 mg, 1.55 mmol) was added diisopropyl
azodicarboxylate (320 ml, 1.55 mmol) at rt, and the resulting
mixture was stirred overnight. The mixture was concentrated in
vacuo, then the residue was chromatographed on silica gel with
hexane-EtOAc (3:1) as eluent to give methyl
3-[2-(S)--(N-Boc-5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate
(513 mg, 87%) as a colorless oil: .sup.1H-NMR (CDCl.sub.3) .delta.
1.40 and 1.46 (s, total 9H), 1.95-2.20 (series of m, 4H), 3.33-3.72
(series of m, 2H), 3.82-4.00 (m, 1H), 3.90 and 3.91 (s, total 3H),
4.09-4.21 (m, 3H), 4.21-4.57 (m, 2H), 7.11-7.15 (m, 1H), 7.26-7.37
(m, 6H), 7.53-7.65 (m, 2H); MS (ESI) m/z, 456 (M.sup.++H).
[0858] To a stirred solution of methyl
3-[2-(S)--N-Boc-5-(S)-benzyloxymethyl)pyrrolidinyl-methoxy]benzoate
(501 mg, 1.10 mmol) in CH.sub.2Cl.sub.2 (10 ml) was added
trifluoroacetic acid (10 ml) at rt, and the resulting mixture was
stirred for 1 h. The mixture was concentrated in vacuo and poured
into aq.NaHCO.sub.3, the extracted with CHCl.sub.3. The organic
layer was washed with water, drying over anhydrous
Na.sub.2SO.sub.4, and concentrated in vacuo to give methyl
3-[2-(S)-5-(S)-benzyloxymethyl)pyrrolidinyl-methoxy]benzoate (387
mg, quant.) as yellowish oil. The product was used for next
reactions without further purification. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.26-1.65 (m, 2H), 1.94-2.04 (m, 3H), 3.37-3.52 (m, 2H),
3.63-3.66 (m, 1H), 3.85-3.93 (m, 1H), 3.91 (s, 3H), 4.55 (s, 2H),
7.09-7.11 (m, 1H), 7.27-7.54 (m, 6H), 7.54-7.55 (m, 1H), 7.61-7.63
(m, 2H); MS (ESI) m/z, 35 (M.sup.++H).
[0859] To a stirred solution of methyl
3-[2-(S)-5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (140
mg, 0.39 mmol), 4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetic
acid (125 mg, 0.39 mmol) and N,N-dimethylaminopyridine (58.0 mg,
0.47 mmol) in THF (15 ml) was added EDC.HCl (90.0 mg, 0.47 mmol) at
rt, and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with CHCl.sub.3. The
organic layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give methyl
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenyl-
acetyl]-2-(S)-pyrrolidinyl methoxy]benzoate (256 mg, quant.) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3), mixture of
rotamars .delta. 1.67 (s, 3H), 1.97-2.40 (series of m, 4H),
3.42-3.85 (series of m, 5H), 3.60 (s, 3H), 3.95 and 3.97 (s, total
3H), 4.15-4.26 (m, 2H), 4.36-4.49 (m, 1H), 4.59 (s, 2H), 6.32 and
6.36 (s, total 1H), 6.75-6.87 (series of m, 2H), 7.20 (brs, 2H),
7.16-7.72 (series of m, 10H), 8.03-8.09 (m, 1H); MS (ESI) m/z, 652
(M.sup.++H).
[0860] To a stirred solution of methyl
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenyl-
acetyl]-2-(S)-pyrrolidinylmethoxy]benzoate (185 mg, 0.28 mmol) in
MeOH-THF (2:5, 7 ml) was added 1.0M-NaOH (860 ml, 0.86 mmol) at rt,
and the resulting mixture was heated at 60.degree. C. with stirring
for 1 h. The reaction mixture was poured into 1N-HCl, then
extracted with CHCl.sub.3. The organic layer was washed with brine,
drying over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(5:1) as eluent to give 129 (171 mg, 94%) as a colorless amorphous
solid. MW 637.72 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars
.delta. 1.89-2.37 (series of m, 4H), 2.29 (s, 3H), 3.52-4.53
(series of m, 8H), 3.66 and 3.74 (s, total 3H), 4.85 (s, 2H),
6.66-7.98 (series of m, 16H); MS (ESI) m/z, 638 (M.sup.++H), 660
(M.sup.++Na.sup.+); Anal. Calcd for
C.sub.37H.sub.39N.sub.3O.sub.7.H.sub.2O: C, 67.77; H, 6.30; N,
6.41. Found: C, 67.40; H, 5.95; N, 6.14.
Example 122
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyla-
cetyl]-2-(S)-pyrrolidinylmethoxy]benzoic acid
[0861] ##STR1751##
[0862] To a stirred solution of methyl
3-[2-(S)-5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (118
mg, 0.33 mmol), 4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic
acid (112 mg, 0.33 mmol) and N,N-dimethylaminopyridine (50.0 mg,
0.40 mmol) in THF (15 ml) was added EDC.HCl (80.0 mg, 0.40 mmol) at
rt, and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with CHCl.sub.3. The
organic layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give methyl
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl-
acetamido]-2-(S)-pyrrolidinylmethoxy]benzoate (226 mg, quant.) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3), mixture of
rotamars .delta. 1.99-2.40 (series of m, 4-H), 3.43-3.92 (series of
m, 5H), 3.67 (s, 3H), 3.98 and 4.02 (s, total 3H), 4.18-4.29 (m,
2H), 4.36-4.51 (m, 1H), 4.60 (s, 2H), 6.75-0.92 (series of m, 2H),
7.01-7.22 (series of m, 4H), 7.29-7.53 (series of m, 9H), 7.62-8.26
(series of m, 3H); MS (ESI) m/z, 672 (M.sup.++H).
[0863] To a stirred solution of methyl
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl-
acetyl]-2-(S)-pyrrolidinylmethoxy]benzoate (169 mg, 0.25 mmol) in
MeOH-THF (2:5, 7 ml) was added 1.0M-NaOH (760 ml, 0.76 mmol) at rt,
and the resulting mixture was heated at 60.degree. C. with stirring
for 1.5 h. The reaction mixture was poured into 1N-HCl, then
extracted with CHCl.sub.3. The organic layer was washed with brine,
dryed over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(5:1) as eluent to give 130 (155 mg, 94%) as a colorless amorphous
solid. MW 658.14 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars
.delta. 1.89-2.35 (series of m, 4H), 3.52-4.53 (series of m, 8H),
3.68 and 3.75 (s, total 3H), 4.85 (s, 2H), 6.68-8.03 (series of m,
16H); MS (ESI) m/z, 658 (M.sup.++H), 680 (M.sup.++Na.sup.+); Anal.
Calcd for C.sub.36H.sub.36ClN.sub.3O.sub.7H.sub.2O: C, 63.95; H,
5.66; N, 6.21. Found: C, 64.01; H, 5.38; N, 5.96.
Example 123
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylac-
etyl]-2-(S)-pyrrolidinylmethoxy]benzoic acid
[0864] ##STR1752##
[0865] To a stirred solution of methyl
3-[2-(S)-(5-(S)-benzyloxymethyl)pyrrolidinylmethoxy]benzoate (116
mg, 0.33 mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic
acid (124 mg, 0.33 mmol) and N,N-dimethylaminopyridine (48.0 mg,
0.39 mmol) in THF (15 ml) was added EDC.HCl (75.0 mg, 0.39 mmol) at
rt, and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with CHCl.sub.3. The
organic layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give methyl
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyla-
cetyl]-2-(S)-pyrrolidinyl methoxy]benzoate (209 mg, 89%) as a
colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3), mixture of
rotamars .delta. 1.99-2.37 (series of m, 4H), 3.43-3.91 (series of
m, 5H), 3.70 (s, 3H), 3.93 and 3.96 (s, total 3H), 4.19-4.28 (m,
2H), 4.37-4.51 (m, 1H), 4.60 (s, 2H), 6.77-7.11 (series of m, 6H),
7.28-7.74 (series of m, 10H), 7.91-7.95 (series of m, 1H),
8.20-8.23 (series of m, 1H); MS (ESI) m/z 716 (NV), 718
(M.sup.++2).
[0866] To a stirred solution of methyl
3-[5-(R)-benzyloxymethyl-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyla-
cetyl]-2-(S)-pyrrolidinylmethoxy]benzoate (176 mg, 0.25 mmol) in
MeOH-THF (2:5, 7 ml) was added 1.0M-NaOH (760 ml, 0.76 mmol) at rt,
and the resulting mixture was heated at 60.degree. C. with stirring
for 1.5 h. The reaction mixture was poured into 1N-HCl, then
extracted with CHCl.sub.3. The organic layer was washed with brine,
drying over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(5:1) as eluent to give 131 (156 mg, 88%) as a colorless amorphous
solid. MW 702.59 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars
.delta. 1.89-2.37 (series of m, 4H), 2.29 (s, 3H), 3.52-4.53
(series of m, 8H), 3.68 and 3.75 (s, total 3H), 4.85 (s, 2H),
6.67-7.95 (series of m, 16H); MS (ESI) m/z, 702 (M.sup.++H), 704
(M.sup.++Na.sup.+); Anal. Calcd for
C.sub.36H.sub.36BrN.sub.3O.sub.7.H.sub.2O: C, 60.00; H, 5.32; N,
5.83. Found: C, 59.65; H, 5.02; N, 5.65.
Example 124
4-[(2S,4S)-4-methoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacety-
l]-2-pyrrolidinyl]methoxybenzoic acid
[0867] ##STR1753##
[0868] To a stirred mixture of
(2S,4R)-1-tert-butoxycarbonyl-2-tert-butyldiphenylsilyloxymethyl-4-hydrox-
ypyrrolidine (21.7 g, 47.6 mmol), acetic acid (3.0 ml, 52.4 mmol)
and PPh.sub.3 (12.5 g, 52.4 mmol) in THF (330 ml) was added DIAD
(9.4 ml, 47.6 mmol) at room temperature under an atmosphere of
nitrogen. After 2 h stirring the same temperature, the mixture was
heated at 50.degree. C. for 2 h. After cooling to room temperature,
the reaction mixture was concentrated in vacuo. The resulting
residue was chromatographed on silica gel [1 Kg, n-hexane/EtOAc
(5/1)], to give
(2S,4S)-4-acetoxy-1-tert-butoxycarbonyl-2-tert-butyldiphenylsilyloxymethy-
pyrrolidine (23.3 g, 99%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.06 (s, 9H), 1.35 and 1.43 (s, 9H, amide
isomers), 1.92 (br, 3H), 2.20-2.45 (m, 2H), 3.31-4.07 (m, 5H),
5.17-5.30 (m, 1H), 7.36-7.44 (m, 6H), 7.65-7.71 (m, 4H).
[0869] To a stirred mixture of
(2S,4S)-4-acetoxy-1-tert-butoxycarbonyl-2-tertbutyldiphenyl
silyloxy methypyrrolidine (23.3 g, 46.9 mmol) and acetic acid (6.0
ml, 104.8 mmol) in THF (470 ml) was added TBAF (93.8 ml, 93.8 mmol)
at 0.degree. C. After 24 h stirring, the mixture was concentrated
in vacuo. The resulting residue was diluted with EtOAC and aq.
NH.sub.4Cl and extracted with EtOAc. The combined extracts were
washed with brine, which were dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was chromatographed on silica
gel [700 g, CHCl.sub.3/EtOAc (4/1)], to give
(2S,4S)-4-acetoxy-1-tert-butoxycarbonyl-2-pyrrolidinemethanol (9.70
g, 8%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47
(s, 9H), 1.63 (m, 1H), 1.81 (m, 1H), 2.07 (s, 3H), 2.34 (m, 1H),
3.42 (dd, J=12.7, 0.9 Hz, 1H), 3.62-3.85 (m, 3H), 4.48 (br, 1H),
5.20 (br, 1H).
[0870] To a stirred mixture of
(2S,4S)-4-acetoxy-1-tert-butoxycarbonyl-2-pyrrolidinemethanol (9.70
g, 37.4 mmol), p-hydroxybenzoic acid methyl ester (5.69 g, 37.4
mmol) and PPh.sub.3 (10.8 g, 41.1 mmol) in THF (200 ml) was added
DIAD (8.10 ml, 41.1 mmol) at room temperature. After 1.5 h
stirring, the mixture was concentrated in vacuo. The resulting
residue was chromatographed on silica gel [700 g, CHCl.sub.3/EtOAc
(10/1).sub.j, to give methyl
4-[(2S,4S)-4-acetoxy-1-tert-butoxycarbonyl-2-pyrrolidinyl]methoxybenzoate
(11.8 g, 81%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.48 (s, 9H), 2.03 (s, 3H), 2.27 (m, 2H), 3.46 (m, 1H),
3.72 (m, 1H), 3.88 (s, 3H), 3.98 (t, J=9.0 Hz, 1H), 4.21-4.47 (m,
2H), 5.31 (br, 1H), 6.96 (br, 2H), 7.98 (d, J=8.8 Hz, 2H).
[0871] To a stiffed solution of methyl
4-[(2S,4S)-4-acetoxy-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinyl]metho-
xybenzoate (7.43 g, 18.9 mmol) in MeOH (150 ml) was added cat.
K.sub.2CO.sub.3 at room temperature. After 1 day stirring, the
mixture was concentrated in vacuo. The resulting residue was
recrystallized by the addition of CHCl.sub.3-n-hexane, to give
methyl
4-[(2S,4S)-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinyl]methoxybenzoate
(5.76 g, 87%) as a colorless solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.46 (s, 9H), 2.11 (m, 1H), 2.35 (br, 1H), 3.27-3.65 (m,
2H), 3.89 (s, 3H), 4.07-4.54 (m, 4H), 6.96 (d, J=6.9 Hz, 2H), 7.99
(d, J=6.9 Hz, 2H).
[0872] To a stirred solution of methyl
4-[(2S,4S)-1-tert-butoxycarbonyl-4-hydroxy-2-pyrrolidinyl]methoxybenzoate
(2.10 g, 5.98 mmol) in THF (60 ml) was added 60% oil NaH (359 mg,
8.97 mmol) at 0.degree. C. After 15 minutes stirring, MeI (1.20 ml,
8.97 mmol) was added to the mixture was added at same temperature,
and the resulting mixture was allowed to raise to room temperature
for over 1 h. Then 60% oil NaH (359 mg, 8.97 mmol) and MeI (1.20
ml, 8.97 mmol) was added to the reaction mixture at room
temperature and stirred for 14 h. The reaction mixture was poured
into ice water and extracted with CHCl.sub.3. The combined extracts
were washed with aq. NaHCO.sub.3 and brine. After dried over
Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [50 g, n-hexane/EtOAc
(4/1)], to give methyl
4-[(2S,4S)-1-tert-butoxycarbonyl-4-methoxy-2-pyrrolidinyl]methoxybenzoate
(1.32 g, 60%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.48 (s, 9H), 2.05 (m, 1H), 2.29 (d, J=14.2 Hz, 1H), 3.30 (s, 3H),
3.36-4.38 (m, 4H), 6.76 (br, 2H), 7.97 (d, J=8.8 Hz, 2H).
[0873] To a stirred solution of methyl
4-[(2S,4S)-1-tert-butoxycarbonyl-4-methoxy-2-pyrrolidinyl]methoxybenzoate
(2.38 g, 3.61 mmol) in CH.sub.2Cl.sub.2 (46 ml) was added TFA (23
ml) at room temperature. After 14 h stirring, the mixture was
concentrated in vacuo. The residue was diluted by the addition of
CH.sub.2Cl.sub.2 and 1 N NaOH, and extracted with CH.sub.2Cl.sub.2.
The combined extracts were washed with brine, dried over
Na.sub.2SO.sub.4, which was concentrated in vacuo. The residue was
chromatographed on silica gel [50 g, CHCl.sub.3/MeOH (20/1)], to
give methyl 4-[(2S,4S)-4-methoxy-2-pyrrolidinyl]methoxybenzoate
(950 mg, 99%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
2.16 (t, J=5.3 Hz, 1H), 2.72 (s, 1H), 2.95 (d, J=6.8 Hz, 1H), 3.11
(d, J=11.0 Hz, 1H), 3.26 (t, J=1.9 Hz, 3H), 3.52 (br, 1H), 3.84 (d,
J=1.7 Hz, 3H), 3.92 (s, 1H), 4.00 (d, J=4.1 Hz, 2H), 6.88 (m, 2H),
7.94 (m, 2H).
[0874] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (375 mg,
1.19 mmol), methyl
4-[(2S,4S)-4-methoxy-2-pyrrolidinyl]methoxybenzoate (317 mg, 1.19
mmol), EDC.HCl (342 mg, 1.79 mmol), HOBT (242 mg, 1.79 mmol) and
Et.sub.3N (0.83 ml, 5.95 mmol) in DMF (5 ml) was stirred at room
temperature for 13 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/Acetone (10/1) CHCl.sub.3/MeOH
(20/1)], to give methyl
4-[(2S,4S)-4-methoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]-2-pyrrolidinyl]methoxybenzoate (650 mg, 98%) as a pale brown
amorphous solid .sup.1H-NMR (CDCl.sub.3) .delta. 2.03 (m, 1H), 2.31
(s, 3H), 2.32 (m, 1H), 3.29 (d, J=1.0 Hz, 3H), 3.57-3.68 (m, 5H),
3.88 (d, J=1.0 Hz, 3H), 3.99-4.06 (m, 2H), 4.46 (m, 1H), 6.19 (m,
1H), 6.80 (s, 1H), 6.81 (d, J=9.0 Hz, 1H), 6.96-7.19 (m, 4H), 7.29
(m, 2H), 7.50 (d, J=6.7 Hz, 1H), 7.95-8.10 (m, 3H); MS (ESI) m/z
562 (M.sup.++1).
[0875] To a solution of methyl
4-[(2S,4S)-4-methoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]-2-pyrrolidinyl]methoxybenzoate (650 mg, 1.16 mmol) in THF (18.5
ml), 0.25 N NaOH (18.5 ml) was added. After stirring at room
temperature for 12 h, the mixture was acidified with 1 N HCl and
extracted with CHCl.sub.3-MeOH (10/1). The combined extracts were
dried over Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was chromatographed on silica gel [50 g, CHCl.sub.3/MeOH (20/1)] to
give 132 (540 mg, 85%) as a colorless amorphous solid. MW 547.60 IR
(KBr) 3354, 2937, 1709, 1685, 1604, 1533, 1454 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.11 (m, 2H), 2.25 (s, 3H), 3.22
(s, 3H), 3.49-3.78 (m, 4H), 3.82 and 3.86 (s, 3H, amide isomers),
3.87-4.52 (m, 4H), 6.71-7.17 (m, 7H), 7.79 (d, J=8.1 Hz, 1H),
7.86-8.03 (m, 3H), 8.45-8.57 (m, 2H), 12.64 (br, 1H); MS (ESI) m/z
548 (M.sup.++1); Anal. Calcd for
C.sub.30H.sub.33N.sub.3O.sub.7.1Na.1.5H.sub.2O: C, 60.29; H, 6.07;
N, 7.03. Found: C, 59.90; H, 5.59; N, 6.69.
Example 125
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-metho-
xy-2-pyrrolidinyl]methoxybenzoic acid
[0876] ##STR1754##
[0877] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (398 mg,
1.19 mmol), methyl
4-[(2S,4S)-4-methoxy-2-pyrrolidinyl]methoxybenzoate (317 mg, 1.19
mmol), EDC.HCl (342 mg, 1.79 mmol), HOBT (242 mg, 1.79 mmol) and
Et.sub.3N (0.83 ml, 5.95 mmol) in DMF (5 ml) was stirred at room
temperature for 13 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/Acetone (10/1) CHCl.sub.3/MeOH
(20/1)], to give methyl
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-meth-
oxy-2-pyrrolidinyl]methoxybenzoate (600 mg, 87%) as a colorless
amorphous solid .sup.1H-NMR (CDCl.sub.3) .delta. 1.99-2.06 (m, 1H),
2.34 (d, J=13.9 Hz, 1H), 3.30 (s, 3H), 3.59 (d, J=7.4 Hz, 1H), 3.62
(d, J=3.2 Hz, 2H), 3.83 (s, 3H), 3.88 (s, 3H), 4.00-4.18 (m, 3H),
4.42-4.51 (m, 2H), 6.82-7.07 (m, 7H), 7.28 (d, J=8.3 Hz, 1H), 7.35
(dd, J=7.9, 1.5 Hz, 1H), 7.94-8.00 (m, 3H), 8.18 (d, J=8.3 Hz, 1H);
MS (ESI) m/z 582 (M.sup.++1), 584 (M.sup.++3).
[0878] To a solution of methyl
4-[(2S,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-4-methoxy-2-pyrrolidinyl]methoxybenzoate (600 mg, 1.03
mmol) in THF (16 ml), 0.25 N NaOH (16 ml) was added. After stirring
at room temperature for 12 h, the mixture was acidified with 1 N
HCl and extracted with CHCl.sub.3-MeOH (10/1). The combined
extracts were dried over Na.sub.2SO.sub.4 and concentrated in
vacuo. The residue was purified on TLC [CHCl.sub.3/MeOH (10/1)] to
give 133 (495 mg, 75%) as a colorless amorphous solid. MW 568.02 IR
(KBr) 3330, 3070, 2937, 1709, 1685, 1604, 1533 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.11 (m, 2H), 3.22 (s, 3H),
3.56-3.78 (m, 4H), 3.81 and 3.85 (s, 3H, amide isomers), 3.88-4.56
(m, 4H), 6.73 and 6.77 (d, J=8.1 Hz, 1H, amide isomers), 6.85 and
6.91 (s, 1H, amide isomers), 7.01-7.07 (m, 3H), 7.28 (t, J=8.1 Hz,
1H), 7.43 (d, J=8.1 Hz, 1H), 7.85-7.94 (m, 2H), 7.97 (d, J=8.6 Hz,
1H), 8.09 (d, J=8.3 Hz, 1H), 8.90-8.95 (m, 2H); MS (FAB) m/z 570
(M.sup.++1), 572 (M.sup.++3).
Example 126
4-[(2S,4S)-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-methox-
y-2-pyrrolidinyl]methoxybenzoic acid
[0879] ##STR1755##
[0880] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (451 mg,
1.19 mmol), methyl
4-[(2S,4S)-4-methoxy-2-pyrrolidinyl]methoxybenzoate (317 mg, 1.19
mmol), EDC.HCl (342 mg, 1.79 mmol), HOBT (242 mg, 1.79 mmol) and
Et.sub.3N (0.83 ml, 5.95 mmol) in DMF (5 ml) was stirred at room
temperature for 13 h. The mixture was poured into ice water and
extracted with EtOAc. The combined extracts were washed with ice
water and brine. After dried over Na.sub.2SO.sub.4, the extracts
were concentrated in vacuo. The residue was chromatographed on
silica gel [50 g, CHCl.sub.3/Acetone (10/1)], to give methyl
4-[(2S,4S)-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy
phenylacetyl]-4-methoxy-2-pyrrolidinyl]methoxybenzoate (760 mg,
100%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.99-2.32
(m, 1H), 2.34 (d, J=13.4 Hz, 1H), 3.30 (s, 3H), 3.59 (m, 1H), 3.63
(d, J=3.2 Hz, 2H), 3.68 (dd, J=12.2, 5.1 Hz, 1H), 3.81 (br, 3H),
3.88 (s, 3H), 3.91-4.16 (m, 2H), 4.49-4.51 (m, 2H), 6.82-7.15 (m,
7H), 7.31 (t, J=8.1 Hz, 1H), 7.52 (d, J=8.1 Hz, 1H), 7.93-8.00 (m,
3H), 8.14 (d, J=8.3 Hz, 1H); MS (ESI) m/z 626 (M.sup.++1), 628
(M.sup.++3).
[0881] To a solution of methyl
4-[(2S,4S)-1-[4-[N'-(2-bromophenyl)ureido]phenylacetyl]-4-methoxy-2-pyrro-
lidinyl]methoxybenzoate (760 mg, 1.19 mmol) in THF (19 ml), 0.25 N
NaOH (19 ml) was added. After stirring at room temperature for 12
h, the mixture was acidified with 1 N HCl and extracted with
CHCl.sub.3-MeOH (10/1). The combined extracts were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
chromatographed on silica gel [50 g, CHCl.sub.3/MeOH (20/1)] to
give 134 (580 mg, 78%) as a colorless amorphous solid. MW 612.47 IR
(KBr) 3330, 2935, 1709, 1685, 1604, 1529, 1434 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.11 (m, 2H), 3.22 (s, 3H),
3.58-3.78 (m, 4H), 3.81 and 3.86 (s, 3H, amide isomers), 3.92-4.52
(m, 4H), 6.72 (d, J=8.6 Hz, 1H), 6.77 (d, J=8.3 Hz, 1H), 6.85 and
6.91 (s, 1H, amide isomers), 6.97 (t, J=7.1 Hz, 1H), 7.02 and 7.06
(d, J=8.6 Hz, 2H, amide isomers), 7.32 (t, J=7.3 Hz, 1H), 7.59 (dd,
J=8.1, 1.0 Hz, 1H), 7.94 (dd, J=8.1, 1.2 Hz, 2H), 7.95-7.98 (m,
2H), 8.74 (s, 1H), 8.94 (s, 1H), 12.63 (br, 1H); MS (FAB) m/z 612
(M.sup.++1), 614 (M.sup.++3).
Example 127
4-[(4R)-methoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2-
S)-pyrrolidinyl methoxy]benzoic acid
[0882] ##STR1756##
[0883] To a stirred solution of
1-(tert-butoxycarbonyl)-(4R)-methoxy-(2S)-pyrrolidinylcarboxylic
acid (2.87 g, 11.7 mmol) in THF (25 ml) was added BH.sub.3DMS (1.66
ml, 17.5 mmol) at room temperature and the reaction mixture was
stirred at room temperature overnight. The mixture was evaporated
and the residue was dissolved with CH.sub.2Cl.sub.2. The solution
was washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4, and
evaporated. The residue was purified by column chromatography on
silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as eluent to give
1-(tert-butoxycarbonyl)-(4R)-methoxy-(2S)-prolinol (1.79 g, 66%) as
a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.47 (s, 9H),
1.69-1.73 (m, 1H), 2.12-2.17 (m, 1H), 3.31 (s, 3H), 3.37-3.40 (m,
1H), 3.53-3.62 (m, 2H), 3.68-3.73 (m, 1H), 3.83-3.87 (m, 1H),
4.04-4.07 (m, 1H), 4.90-4.92 (m, 1H); MS (FAB) m/z 232
(M.sup.++1).
[0884] To a stirred solution of methyl 4-hydroxybenzoate (1.18 g,
7.76 mmol), 1-(tert-butoxy carbonyl)-(4R)-methoxy-(2S)-prolinol
(1.79 g, 7.74 mmol) and Ph.sub.3P (2.44 g, 9.30 mmol) in THF (30
ml) was added DIAD (1.83 ml, 9.29 mmol) and the reaction mixture
was heated under reflux for 5 hr. After cooled to room temperature,
the mixture was evaporated. The residue was filtered on silica-gel
with toluene-acetone (5:1, v/v) as eluent to give the crude
product. The crude product was dissolved in CH.sub.2Cl.sub.2 (20
ml). The solution was added TFA (20 ml) and the reaction mixture
was stirred at room temperature overnight. The mixture was
concentrated in vacuo and made basic by sat. NaHCO.sub.3. The
mixture was extracted with CHCl.sub.3, washed with brine, dried
over K.sub.2CO.sub.3, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (30:1 to
30:2, v/v) as eluent to give methyl
4-[(4R)-methoxy-(2S)-pyrrolidinyl methoxy]benzoate (1.67 g, 81% for
2 steps) as a reddish brown oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.65-1.72 (m, 1H), 1.89 (bs, 1H), 2.05-2.22 (m, 1H), 2.95-3.15 (m,
2H), 3.31 (s, 3H), 3.69-3.76 (m, 1H), 3.88 (s, 3H), 3.91-4.06 (m,
3H), 6.89-6.92 (m, 2H), 7.96-7.98 (m, 2H); MS (FAB) m/z 266
(M.sup.++1).
[0885] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (470 mg,
1.50 mmol), methyl
4-[(4R)-methoxy-(2S)-pyrrolidinylmethoxy]benzoate (396 mg, 1.49
mmol), EDC.HCl (343 mg, 1.79 mmol), HOBt (242 mg, 1.79 mmol) and
Et.sub.3N (250 ml, 1.79 mmol) in THF (10 ml) was stirred at room
temperature overnight. The mixture was diluted with H.sub.2O and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1,
v/v) as eluent to give methyl
4-[(4R)-methoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-(2S)-pyrrolidinylmethoxy]benzoate (822 mg, 98%) as a white
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.14-2.24 (m, 2H), 2.27 (s,
3H), 3.25 (s, 3H), 3.51 (s, 3H), 3.58-3.73 (m, 4H), 3.88 (s, 3H),
3.98-4.09 (m, 2H), 4.40-4.53 (m, 2H), 6.67-7.29 (series of m, total
9H), 7.57-7.59 (m, 1H), 7.91-7.93 (m, 2H), 8.04-8.06 (m, 1H); MS
(FAB) m/z 562 (M.sup.++1).
[0886] To a stirred solution of methyl
4-[(4R)-methoxy-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(-
2S)-pyrrolidinylmethoxy]benzoate (517 mg, 0.92 mmol) in THF (5 ml)
was added 0.5 N NaOH (5 ml) and the reaction mixture was heated
under reflux for 3 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl and the resulting precipitate
was collected under a reduced pressure. The crude solid was
purified by recrystallization from MeOH--CHCl.sub.3--IPE to give
135 (144 mg, 29%) as a white crystalline powder. MW 547.60 mp
112-115.degree. C.; .sup.1H-NMR (DMSO) .delta. 2.04-2.17 (m, 2H),
2.25 (s, 3H), 3.21 (s, 3H), 3.56-3.75 (m, 4H), 3.79 (s, 3H),
4.04-4.35 (m, 4H), 6.73-7.17 (series of m, total 7H), 7.79-7.81 (m,
1H), 7.87-7.89 (m, 2H), 7.99-8.01 (m, 1H), 8.47 (s, 1H), 8.55 (s,
1H), 12.63 (bs, 1H); MS (FAB) m/z 548 (M.sup.++1); Anal. Calcd for
C.sub.30H.sub.33N.sub.3O.sub.7.1/4H.sub.2O: C, 65.26; H, 6.12; N,
7.61. Found: C, 65.36; H, 6.45; N, 7.24.
Example 128
4-[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-methoxy-(2-
S)-pyrrolidinyl methoxy]benzoic acid
[0887] ##STR1757##
[0888] A mixture of
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetic acid (476 mg,
1.50 mmol), methyl 4-[(4R)-methoxy(2S)-pyrrolidinylmethoxy]benzoate
(397 mg, 1.50 mmol), EDC.HCl (344 mg, 1.79 mmol), HOBt (243 mg,
1.80 mmol) and Et.sub.3N (250 ml, 1.79 mmol) in THF (10 ml) was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-methoxy-(-
2S)-pyrrolidinylmethoxy]benzoate (806 mg, 95%) as a pale yellow
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.14-2.37 (m, 2H), 3.28 (s,
3H), 3.44 (s, 3H), 3.48-3.74 (m, 4H), 3.88 (s, 3H), 4.02-4.15 (m,
2H), 4.43-4.58 (m, 2H), 6.63-7.10 (series of m, total 7H),
7.68-7.73 (m, 1H), 7.89-8.02 (m, 4H), 8.16-8.20 (m, 1H); MS (FAB)
m/z 566 (M.sup.++1).
[0889] To a stirred solution of methyl
4-[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenyl
acetyl]-(4R)-methoxy-(2S)-pyrrolidinylmethoxy]benzoate (491 mg,
0.87 mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the
reaction mixture was heated under reflux for 3 hr. After cooled to
room temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected under a reduced pressure. The
crude solid was purified by recrystallization from
MeOH--CHCl.sub.3--IPE to give 136 (173 mg, 36%) as a white
crystalline powder. MW 551.56 mp 111-116.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.08-2.17 (m, 2H), 3.21 (s, 3H), 3.56-3.73
(m, 4H), 3.78 (s, 3H), 4.04-4.33 (m, 4H), 6.74-7.22 (series of m,
total 7H), 7.87-7.89 (m, 2H), 7.99-8.01 (m, 1H), 8.16-8.20 (m, 1H),
8.70 (s, 1H), 9.18 (s, 1H), 12.64 (br s, 1H); MS (FAB) m/z 552
(M.sup.+30 1); Anal. Calcd for
C.sub.29H.sub.30FN.sub.3O.sub.7.0.15H.sub.2O: C, 62.84; H, 5.51; F,
3.43; N, 7.58. Found: C, 63.08; H, 5.83; F, 3.30; N, 7.15.
Example 129
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-methoxy-(2-
S)-pyrrolidinyl methoxybenzoic acid
[0890] ##STR1758##
[0891] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (460 mg,
1.37 mmol), methyl
4-[(4R)-methoxy-(2S)-pyrrolidinylmethoxy]benzoate (365 mg, 1.38
mmol), EDC.HCl (316 mg, 1.65 mmol), HOBt (223 mg, 1.65 mmol) and
Et.sub.3N (230 ml, 1.65 mmol) in THF (10 ml) was stirred at room
temperature overnight The mixture was diluted with H.sub.2O and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (100:1, v/v) as
eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-methoxy-(-
2S)-pyrrolidinylmethoxy]benzoate (801 mg, q.y.) as a white foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 2.13-2.36 (m, 2H), 3.27 (s, 3H),
3.58 (s, 3H), 3.61-3.73 (m, 4H), 3.88 (s, 3H), 4.06-4.14 (m, 2H),
4.43-4.56 (m, 2H), 6.70-6.99 (series of m, total 5H), 7.23-7.42 (m,
4H), 7.90-8.00 (m, 3H), 8.17-8.20 (m, 1H); MS (FAB) m/z 582
(M.sup.++1).
[0892] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-(4R)-methoxy-(2S)-pyrrolidinylmethoxy]benzoate (541 mg,
0.93 mmol) in THF (5 ml) was added 0.5 N NaOH (5 ml) and the
reaction mixture was heated under reflux for 3 hr. After cooled to
room temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected under a reduced pressure. The
crude solid was purified by recrystallization from
MeOH--CHCl.sub.3--IPE to give 137 (281 mg, 53%) as a white
crystalline powder. MW 568.02 mp 116-119.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.08-2.17 (m, 2H), 3.21 (s, 3H), 3.56-3.73
(m, 4H), 3.79 (s, 3H), 4.04-4.33 (m, 4H), 6.75 (d, J=8.3 Hz, 1H),
6.87 (s, 1H), 7.02 (d, J=8.3 Hz, 3H), 7.28 (t, J=7.8 Hz, 1H), 7.44
(d, J=7.8 Hz, 1H), 7.87-7.89 (m, 2H), 7.96 (d, J=8.3 Hz, 1H), 8.10
(d, J=8.3 Hz, 1H), 8.89 (s, 1H), 8.93 (s, 1H), 12.63 (br s, 1H); MS
(FAB) m/z 568 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.30ClN.sub.3O.sub.7.1/4H.sub.2O: C, 60.84; H, 5.37;
Cl, 6.19; N, 7.34. Found: C, 61.03; H, 5.56; Cl, 6.27; N, 7.03.
Example 130
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-methoxy-(2S-
)-pyrrolidinyl methoxy]benzoic acid
[0893] ##STR1759##
[0894] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (600 mg,
1.58 mmol), methyl
4-[(4R)-methoxy-(2S)-pyrrolidinylmethoxy]benzoate (420 mg, 1.58
mmol), EDC.HCl (364 mg, 1.90 mmol), HOBt (214 mg, 1.58 mmol) and
Et.sub.3N (265 ml, 1.90 mmol) in THF (15 ml) was stirred at room
temperature overnight. The mixture was diluted with H.sub.2O and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1,
v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(4R)-methoxy(2S-
)-pyrrolidinylmethoxy]benzoate (1.01 g, q.y.) as a pale yellow
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.13-2.33 (m, 2H), 3.27 (s,
3H), 3.57 (s, 3H), 3.61-3.72 (m, 4H), 3.88 (s, 3H), 4.05-4.14 (m,
2H), 4.43-4.57 (m, 2H), 6.70-7.00 (series of m, total 5H),
7.29-7.52 (m, 4H), 7.92-8.01 (m, 3H), 8.12-8.15 (m, 1H); MS (FAB)
m/z 626 (M.sup.++1).
[0895] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(4R)-methoxy-(2S)-pyrrolidinylmethoxy]benzoate (697 mg,
1.11 mmol) in THF (8 ml) was added 0.5 N NaOH (8 ml) and the
reaction mixture was heated under reflux for 2 hr. After cooled to
room temperature, the mixture was poured into ice-1 N HCl and the
resulting precipitate was collected under a reduced pressure. The
crude solid was purified by recrystallization from
MeOH--CHCl.sub.3--IPE to give 138 (252 mg, 37%) as a white
crystalline powder. MW 612.47 mp 125-130.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.08-2.17 (m, 2H), 3.21 (s, 3H), 3.60-3.72
(m, 4H), 3.79 (s, 3H), 3.95-4.33 (m, 4H), 6.75-7.08 (series of m,
total 5H), 7.31-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.89 (m, 2H),
7.93-7.96 (m, 2H), 8.73 (s, 1H), 8.91 (s, 1H), 12.63 (br s, 1H); MS
(FAB) m/z 612 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.30BrN.sub.3O.sub.7: C, 56.87; H, 4.94; Br, 13.05; N,
6.86. Found: C, 56.67; H, 4.97; Br, 13.07; N, 6.68.
Example 131
4-[4,4-difluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2--
pyrrolidinylmethoxy]benzoic acid
[0896] ##STR1760##
[0897] To a stirred solution of N-Boc proline methyl ester (2.0 g,
8.15 mmol) in CH.sub.2Cl.sub.2 were added 3 .ANG. molecular sieves
(2 g) and PDC (4.60 g, 12.2 mmol). The mixture was stirred for 3
days. The mixture was filtered through a Celite pad and the
filtrate was evaporated. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (10:1) as eluent to give methyl
1-(tert-butoxycarbonyl)-4-oxopyrrolidine-2-carboxylate (1.13 g,
57%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.46-1.48
(m, 9H), 2.56-2.61 (m, 1H), 2.88-3.00 (m, 1H), 3.77 (s, 3H),
3.82-3.88 (m, 2H), 4.71-4.83 (m, 1H).
[0898] To a cold (-78.degree. C.), stirred solution of methyl
1-(tert-butoxycarbonyl)-4-oxopyrrolidine-2-carboxylate (1.13 g,
4.65 mmol) in CH.sub.2Cl.sub.2 (20 ml) was added methylDAST (1.1
ml, 11.6 mmol). The mixture was allowed to warm to room
temperature. After 15 h stirring, the mixture was poured into
H.sub.2O (50 ml) and extracted with EtOAc (200 ml). The extract was
washed with brine (2.times.200 ml), dried over MgSO.sub.4, and
evaporated. The residue was chromatographed on silica gel with
CHCl.sub.3-EtOAc (20:1) as eluent to give methyl
1-(tert-butoxycarbonyl)-4,4-difluoropyrrolidine-2-carboxylate (885
mg, 72%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.42 and
1.47 (s, each, total 9H), 2.46 (ddd, d=26.9 13.7, 5.1 Hz, 1H),
2.62-2.78 (m, 1H), 3.75-3.95 (m, 5H), 4.43-4.57 (m, 1H).
[0899] To a stirred solution of methyl
1-(tert-butoxycarbonyl)-4,4-difluoropyrrolidine-2-carboxylate (885
mg, 3.34 mmol) in THF (25 ml) was added 0.25 N NaOH (26.7 ml, 6.67
mmol) and the stirring was continued for 1 h. The mixture was
poured into 1 N HCl (100 ml) and extracted with CHCl.sub.3
(2.times.200 ml). The combined extracts were washed with brine (100
ml), dried over MgSO.sub.4, and evaporated to give
1-(tert-butoxycarbonyl)-4,4-difluoropyrrolidine-2-caboxylic acid
(775 mg, 92%) as a yellow crystalline solid. mp 113-117.degree. C.;
.sup.1H-NMR (CDCl.sub.3) .delta. 1.44 and 1.49 (s, each, total 9H),
2.53-2.80 (m, 2H), 3.71-3.90 (m, 2H), 4.20-4.61 (m, 1H); MS (FAB)
m/z, 252 (M.sup.++1); Anal. Calcd for
C.sub.10H.sub.15F.sub.2O.sub.4: C, 47.81; H, 6.02; N, 5.58. Found:
C, 48.06; H, 6.05; N, 5.45.
[0900] To a stirred solution of N-(tert-butoxycarbonyl)
4,4-difluoroproline (3.00 g, 11.9 mmol) in THF (20 ml) was added
BH.sub.3 DMS (1.1 ml, 11.9 mmol) at room temperature. The mixture
was heated at reflux for 2 h. After cooling to room temperature,
the mixture was concentrated in vacuo. The residue was quenched by
the addition of H.sub.2O (100 ml) and extracted with CHCl.sub.3
(2.times.200 ml). The combined extracts were dried over MgSO.sub.4
and evaporated. The residue was chromatographed on silica gel with
CHCl.sub.3-EtOAc (4:1) as eluent to give
1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethanol (2.11
g, 75%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.48
(s, 9H), 2.04-2.55 (m, 2H), 3.59-4.17 (m, 5H).
[0901] To a stirred mixture of
1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethanol (600
mg, 2.53 mmol), methyl 4-hydroxybenzoate (462 mg, 3.03 mmol),
Ph.sub.3P (795 mg, 3.03 mmol) in THF (10 ml) was added DIAD (597
ul, 3.03 mmol) at room temperature. The mixture was heated at
reflux for 3 h with stirring. After cooling to room temperature,
the mixture was concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-EtOAc (4:1) as eluent to
give methyl
4-[1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethoxy]benzoate
(831 mg, 88%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.48 (s, 9H), 2.53-2.61 (m, 2H), 3.63-4.41 (series of m, total 8H),
6.94 (d, J=8.8 Hz, 2H), 7.99 (d, J=8.8 Hz, 2H).
[0902] A mixture of methyl
4-[1-(tert-butoxycarbonyl)-4,4-difluoro-2-pyrrolidinylmethoxy]benzoate
(830 mg, 2.23 mmol) and TFA (5 ml) in CH.sub.2Cl.sub.2 (5 ml) was
stirred for 3 h and concentrated in vacuo. The residue was made
basic with sat. NaHCO.sub.3 and extracted with CHCl.sub.3
(2.times.200 ml). The combined extracts were dried over
K.sub.2CO.sub.3 and concentrated in vacuo to give methyl
4-(4,4-difluoro-2-pyrrolidinylmethoxy)benzoate (550 mg, 91%) as a
pale yellow solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.19 (m, 1H),
2.43 (m, 1H), 3.19-3.41 (m, 2H), 3.77 (m, 1H), 3.89 (s, 3H),
4.00-4.09 (m, 2H), 6.92 (d, J=9.0 Hz, 2H), 7.99 (d, J=9.0 Hz, 2H);
MS (FAB) m/z 272 (M.sup.++1); Anal. Calcd for
C.sub.13H.sub.15F.sub.2NO.sub.3: C, 57.56; H, 5.57; N, 5.16. Found:
C, 57.65; H, 5.67; N, 5.16.
[0903] A mixture of methyl
4-(4,4-difluoro-2-pyrrolidinylmethoxy)benzoate (540 mg, 1.99 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (626 mg,
1.99 mmol), EDC.HCl (572 mg, 2.99 mmol), HOBt (cat.), and DMAP
(cal) in DMF (10 ml) was stirred overnight. The mixture was diluted
with EtOAc (300 ml), washed with brine (2.times.100 ml), dried over
MgSO.sub.4, and concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[4,4-difluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-
-pyrrolidinylmethoxy]benzoate (1.00 g, 89%) as a colorless foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 2.31 (s, 3H), 2.47-2.63 (m, 2H),
3.52-3.97 (series of s and m, total 10H), 4.07-4.30 (m, 2H),
4.67-4.69 (m, 1H), 6.45 (s, 1H), 6.65 (d, J=1.7 Hz, 1H), 6.74-6.76
(m, 1H), 6.84 (d, J=8.8 Hz, 2H), 7.14 (m, 2H), 7.24 (m, 2H),
7.52-7.54 (m, 1H), 7.94 (d, J=8.8 Hz, 2H), 8.09 (d, J=8.1 Hz,
1H).
[0904] A mixture of methyl
4-[4,4-difluoro-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl-acetyl]--
2-pyrrolidinylmethoxy]benzoate (1.00 g, 1.76 mmol) and 0.25 N NaOH
(14 ml, 3.50 mmol) in THF (14 ml) was stirred overnight. The
mixture was acidified with 1 N HCl and extracted with
CHCl.sub.3-MeOH (10:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1 to 10:1)
as eluent to give 139 (658 mg, 68%) as a colorless crystalline
powder. MW 553.55 mp 135-140.degree. C.; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.23 (s, 3H), 2.49-2.73 (m, 2H), 3.36-4.55 (series of m,
10H), 6.73 (d, J=8.3 Hz, 1H), 6.84 (s, 1H), 6.93 (t, J=7.3 Hz, 1H),
7.00 (d, J=8.3 Hz, 2H), 7.10-7.16 (m, 2H), 7.78 (d, J=8.3 Hz, 1H),
7.86 (d, J=8.3 Hz, 2H), 8.00 (d, J=8.3 Hz, 1H), 8.47 (s, 1H), 8.56
(s, 1H); MS (FAB) m/z, 554 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.29F.sub.2N.sub.3O.sub.6.3/4H.sub.2O: C, 61.42; H,
5.44; N, 7.06. Found: C, 61.30; H, 5.44; N, 7.06.
Example 132
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4,4-difluoro-2--
pyrrolidinylmethoxy]benzoic acid
[0905] ##STR1761##
[0906] A mixture of methyl
4-(4,4-difluoro-2-pyrrolidinylmethoxy)benzoate (229 mg, 0.845
mmol), 4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid
(283 mg, 0.845 mmol), EDC.HCl (243 mg, 1.27 mmol), HOBt (cat.),
DMAP (cat), and DMF (10 ml) was stirred overnight. The mixture was
diluted with EtOAc (300 ml). The solution was washed with brine
(2.times.100 ml), dried over MgSO.sub.4, and concentrated in vacuo.
The residue was chromatographed on silica gel with CHCl.sub.3-EtOAc
(20:1 to 4:1) as eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy
phenylacetyl]-4,4-difluoro-2-pyrrolidinylmethoxybenzoate (482 mg,
97%) as a colorless viscous solid. .sup.1H-NMR (CDCl.sub.3) .delta.
2.50-2.67 (m, 2H), 3.54-4.71 (series of m, 13H), 6.69 (d, J=1.5 Hz,
1H), 6.76 (d, J=8.3 Hz, 1H), 6.84 (d, J=8.8 Hz, 2H), 6.98 (dt,
J=7.8, 1.5 Hz, 1H), 7.23-7.27 (m, 1H), 7.33 (d, J=8.3 Hz, 1H), 7.39
(m, 2H), 7.94 (d, J=8.8 Hz, 2H), 8.00 (d, J=8.3 Hz, 1H), 8.19 (dd,
J=8.3, 1.5 Hz, 1H).
[0907] A mixture of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4,4-difluoro-2-
-pyrrolidinylmethoxy]benzoate (480 mg, 0.816 mmol), 0.25 N NaOH
(6.5 ml, 1.65 mmol), and THF (20 ml) was stirred for 3 days. The
mixture was poured into 1 N HCl (100 ml) and extracted with
CHCl.sub.3-MeOH (5:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1 to 5:1) to
give 140 (270 mg, 58%) as a pale yellow amorphous solid. MW 573.97
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.45-2.74 (m, 2H), 3.63-4.83
(series of m, 10H), 6.76 (d, J=8.3 Hz, 1H), 6.87 (s, 1H), 7.00-7.05
(m, 3H), 7.26-7.30 (m, 1H), 7.44 (dd, J=8.3, 1.2 Hz, 1H), 7.88-7.93
(m, 2H), 7.98 (d, J=8.3 Hz, 1H), 8.10 (d, J=8.3 Hz, 1H), 8.92 (s,
1H), 8.96 (s, 1H); MS (FAB) m/z 574 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.26ClF.sub.2N.sub.3O.sub.6.H.sub.2O: C, 56.81; H,
4.77; N, 7.10. Found: C, 56.75; H, 4.69; N, 6.79.
Example 133
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-1-[4-[N'-(2-methylphenyl)ureido]-3-m-
ethoxyphenylacetyl]-2-pyrrolidinyl]methoxybenzoic acid
[0908] ##STR1762##
[0909] To a solution of methyl
(2S,3R,4S)-1-benzyloxycarbonyl-3,4-isopropylidenedioxy-2-pyrrolidinyl
carboxylate (10.7 g, 31.9 mmol) in THF (250 ml), 0.25 N NaOH (255
ml) was added. After stirring at room temperature for 24 h, the
mixture was acidified with 1 N HCl and extracted with EtOAc. The
combined extracts were washed with brine, which were dried over
Na.sub.2SO.sub.4 and concentrated in vacuo, to give
(2S,3R,4S)-1-benzyloxycarbonyl-3,4-isopropylidenedioxy-2-pyrrolidinylcarb-
oxylic acid (9.87 g, 96%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.32 (s, 3H), 1.46 (d, J=2.7 Hz, 3H), 3.61 (m,
1H), 3.82 and 3.92 (d, J=12.7 Hz, 1H, amide isomers), 4.58 and 4.64
(s, 1H, amide isomers), 4.77 (t, J=5.1 Hz, 1H), 4.83 and 4.89 (d,
J=5.9 Hz, 1H, amide isomers), 5.15 and 5.19 (m, 2H, amideisomers),
7.31-7.37 (m, 5H).
[0910] To a stirred solution of
(2S,3R,4S)-1-benzyloxycarbonyl-3,4-isopropylidenedioxy-2-pyrrolidinyl
carboxylic acid (9.87 g, 30.7 mmol) in THF (200 ml) was added
BH.sub.3-DMS (6.14 ml, 61.4 mmol) at 0.degree. C. The mixture was
allowed to room temperature and then heated under reflux for 2 h.
After cooling to room temperature, the mixture was concentrated in
vacuo and quenched by the addition of water at 0.degree. C. The
mixture was extracted with EtOAc. The combined extracts were washed
with water and brine, which were dried over Na.sub.2SO.sub.4 and
concentrated in vacuo. The residue was chromatographed on silica
gel [200 g, CHCl.sub.3/MeOH (20/1)], to give (2R,3R,4S)-1-benzyloxy
carbonyl-3,4-isopropylidenedioxy-2-pyrrolidinylmethanol (10.1 g,
100%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.31 (s,
3H), 1.45 (s, 3H), 3.56-4.74 (m, 7H), 5.14 (s, 2H), 7.34 (m,
5H).
[0911] To a stirred mixture of
(2R,3R,4S)-1-benzyloxycarbonyl-3,4-isopropylidenedioxy-2-pyrrolidinyl
methanol (312 mg, 0.64 mmol), methyl p-hydroxybenzoate (67 ml, 0.70
mmol), PPh.sub.3 (184 mg, 0.70 mmol) in THF (7 ml) was added DIAD
(138 ml, 0.70 mmol) at 0.degree. C. under an atmosphere of
nitrogen. The mixture was allowed to reach room temperature and
stirred for 3 h. After removal of the solvent, the resulting
residue was chromatographed on silica gel 110 g, n-hexane/EtOAc
(4/1)], to give methyl
4-[(2R,3R,4S)-benzyloxycarbonyl-3,4-isopropylidenedioxy-2-pyrrolidinyl]me-
thoxybenzoate (321 mg, 83%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.01 (s, 6H), 1.03 (s, 3H), 2.23 (m, 1H), 2.63
(m, 1H), 3.61 (d, J=12.5 Hz, 1H), 3.80-4.27 (m, 4H), 4.84 (br, 1H),
5.01 and 5.08 (ABq, J=12.2 Hz, 1H, amide isomers), 6.75-6.87 (m,
3H), 7.19-7.63 (m, 15H).
[0912] A suspension of methyl
4-[(2R,3R,4S)-1-benzyloxycarbonyl-3,4-isopropylidenedioxy-2-pyrrolidinyl]-
methoxybenzoate (2.37 g, 5.76 mmol) and 10% Pd/C (240 mg) in EtOH
(170 ml) was stirred at room temperature under an atmosphere of
hydrogen. After 1 day stirring, the catalyst and solvent were
changed for 10% Pd/C (500 mg) and THF (50 ml). The suspension was
stirred at room temperature under an atmosphere of hydrogen for 5
days. After removed the catalyst by filtration, the filtrates were
concentrated in vacuo. The residue was chromatographed on silica
gel [100 g, CHCl.sub.3/acetone (20/1)], to give methyl
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoa-
te (930 mg, 53%) as a brown oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.35 (s, 3H), 1.50 (s, 3H), 3.02 (dd, J=13.7, 4.1 Hz, 1H), 3.13 (d,
J=13.7 Hz, 1H), 3.58 (t, J=6.3 Hz, 1H), 3.88 (s, 3H), 3.90 (dd,
J=9.3, 6.6 Hz, 1H), 4.02 (dd, J=9.5, 3.9 Hz, 1H), 4.74 (d, J=5.6
Hz, 1H), 4.79 (m, 1H), 6.90 (d, J=9.0 Hz, 2H), 7.98 (d, J=9.0 Hz,
2H).
[0913] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (437 mg,
1.39 mmol), methyl
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoate
(428 mg, 1.39 mmol), EDC.HCl (400 mg, 2.09 mmol) and DMAP (170 mg,
1.39 mmol) in DMF (12 ml) was stirred at room temperature for 20 h.
The mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [70 g, CHCl.sub.3/acetone
(10/1)], to give methyl
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-1-[4-[N'-(2-methylphenyl)ureido]-3--
methoxyphenylacetyl]-2-pyrrolidinyl]methoxybenzoate (840 mg, 100%)
as a colorless amorphous solid IR (KBr) 3354, 2985, 2939, 1716,
1533, 1254 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.31 (s,
3H), 1.42 (s, 3H), 2.05 (s, 3H), 3.50 (s, 3H), 3.55-3.88 (m, 4H),
3.89 (s, 3H), 4.13 (m, 1H), 4.67 (br, 1H), 4.78 (d, J=6.1 Hz, 1H),
4.88 (t, J=5.6 Hz, 1H), 6.46 (s, 1H), 6.62 (d, J=1.5 Hz, 1H), 6.74
(m, 3H), 7.05 (s, 1H), 7.14 (d, J=7.3 Hz, 1H), 7.23 (m, 2H), 7.57
(d, J=7.8 Hz, 1H), 7.91-8.08 (m, 3H); MS (ESI) m/z 604 (M.sup.++1);
Anal. Calcd for C.sub.33H.sub.37N.sub.3O.sub.8.0.6H.sub.2O: C,
64.50; H, 6.27; N, 6.84. Found: C, 64.38; H, 6.18; N, 6.66.
[0914] A mixture of methyl
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-1-[4-[N'-(2-methylphenyl)ureido]-3--
methoxyphenylacetyl]-2-pyrrolidinyl]methoxybenzoate (183 mg, 0.303
mmol) and g.HCl-MeOH (6 ml) was stirred at room temperature for 17
h. The mixture was concentrated in vacuo. The residue was purified
on TLC [CHCl.sub.3/MeOH (10/1)], to give methyl
4-[(2R,3R,4S)-3,4-dihydroxy-1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphe-
nylacetyl]-2-pyrrolidinyl]methoxybenzoate (162 mg, 95%) as a
colorless amorphous solid IR (KBr) 3342, 1716, 1604, 1535, 1255
cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 2.25 (br, 3H),
3.33-3.75 (m, 7H), 3.87 (s, 3H), 4.10 (d, J=8.3 Hz, 1H), 4.24 (s,
2H), 4.37 (m, 2H), 6.62-7.94 (m, 13H); MS (ESI) m/z 564
(M.sup.++1).
[0915] To a solution of methyl
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-1-[4-[N'-(2-methylphenyl)ureido]-3--
methoxyphenylacetyl]-2-pyrrolidinyl]methoxybenzoate (490 mg, 0.812
mmol) in THF (9.8 ml), 0.25 N NaOH (9.8 ml) was added. After
stirring at room temperature for 4 days, the mixture was acidified
with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1). The
combined extracts were dried over Na.sub.2SO.sub.4 and concentrated
in vacuo to give 141 (445 mg, 93%) as a colorless amorphous solid.
MW 689.64 IR (KBr) 3354, 2983, 2937, 1707, 1604, 1533 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) 5.24 and 1.26 (s, 3H, amide isomers),
1.26 and 1.32 (s, 3H, amide isomers), 2.24 (s, 3H), 3.40 (dd,
J=14.0, 5.1 Hz, 1H), 3.60 (m, 2H), 3.71 (m, 1H), 3.76 (s, 3H), 3.82
(s, 3H), 3.92-4.96 (m, 5H), 6.74 and 6.78 (m, 1H, amide isomers),
6.83-7.16 (m, 6H), 7.79 (d, J=8.3 Hz, 1H), 7.87 (t, J=9.1 Hz, 2H),
8.01 (m, 1H), 8.49 (d, J=3.4 Hz, 1H), 8.57 (s, 1H); MS (FAB) m/z
590 (M.sup.++1); Anal. Calcd for
C.sub.32H.sub.35N.sub.3O.sub.9.2.3H.sub.2O: C, 60.90; H, 6.32; N,
6.66. Found: C, 61.00; H, 6.00; N, 6.27.
Example 134
4-[(2R,3R,4S)-3,4-hydroxy-1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenyl-
acetyl]-2-pyrrolidinyl]methoxybenzoic acid
[0916] ##STR1763##
[0917] To a solution of methyl
4-[(2R,3R,4S)-3,4-dihydroxy-1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphe-
nylacetyl]-2-pyrrolidinyl]methoxybenzoate (63 mg, 0.112 mmol) in
THF (0.89 ml), 0.25 N NaOH (0.89 ml) was added. After stirring at
room temperature for 3 days, the mixture was acidified with 1 N HCl
and extracted with CHCl.sub.3-MeOH (10/1). The combined extracts
were dried aver Na.sub.2SO.sub.4 and concentrated in vacuo to give
142 (54 mg, 88%) as a colorless amorphous solid. MW 549.57 IR (KBr)
3356, 2958, 2927, 1685, 1604, 1535, 1255 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.24 (s, 3H), 3.40 (m, 1H), 3.58 (s, 2H),
3.66 (dd, J=9.8, 6.6 Hz, 1H), 3.80 (s, 3H), 3.99-4.30 (m, 5H), 5.10
(br, 1H), 6.72 (d, J=8.1 Hz, 1H), 6.85 (s, 1H), 6.93 (t, J=7.3 Hz,
1H), 7.03 (d, J=8.8 Hz, 2H), 7.14 (t, J=8.8 Hz, 2H), 7.79 (d, J=8.1
Hz, 2H), 7.86 (d, J=8.8 Hz, 2H), 7.99 (d, J=8.3 Hz, 1H), 8.46 (s,
1H), 8.56 (s, 1H); MS (ESI) m/z 550 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.8.0.85H.sub.2O: C, 61.66; H, 5.83; N,
7.44. Found: C, 62.09; H, 5.93; N, 6.95.
Example 135
4-[(2R,3R,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4--
isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoic acid
[0918] ##STR1764##
[0919] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (487 mg,
1.45 mmol), methyl
4-[(2R,3R,4S)-3,4-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoate
(447 mg, 1.45 mmol), EDC.HCl (418 mg, 2.18 mmol) and DMAP (177 mg,
1.45 mmol) in DMF (12 ml) was stirred at room temperature for 19 h.
The mixture was poured into ice water and extracted with EtOAc. The
combined extracts were washed with ice water and brine. After dried
over Na.sub.2SO.sub.4, the extracts were concentrated in vacuo. The
residue was chromatographed on silica gel [70 g, CHCl.sub.3/acetone
(10/1)], to give methyl
4-[(2R,3R,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4-
-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoate (850 mg, 94%)
as a colorless amorphous solid IR (KBr) 3329, 2939, 1716, 1627,
1531, 1254 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.33 (s,
3H), 1.43 (s, 3H), 3.56 (s, 3H), 3.61 (s, 1H), 3.64 (s, 1H), 3.70
(m, 1H), 3.79 (d, J=10.4 Hz, 1H), 3.88 (s, 3H), 4.14 (dd, J=9.8,
2.2 Hz, 1H), 4.40 (dd, J=9.8, 3.4 Hz, 1H), 4.67 (s, 1H), 4.80 (d,
J=6.1 Hz, 1H), 4.90 (t, J=4.6 Hz, 1H), 6.65 (d, J=1.7 Hz, 1H),
6.71-6.84 (m, 3H), 6.98 (dt, J=7.6, 1.5 Hz, 1H), 7.27 (m, 2H), 7.33
(dd, J=8.0, 1.2 Hz, 2H), 7.90-8.01 (m, 3H), 8.20 (dd, J=8.3, 1.5
Hz, 1H); MS (ESI) m/z 624 (M.sup.++1), 626 (M.sup.++3); Anal. Calcd
for C.sub.32H.sub.34N.sub.3O.sub.8.1.4H.sub.2O: C, 59.19; H, 5.71;
N, 6.47. Found: C, 58.85; H, 5.35; N, 6.21.
[0920] A mixture of methyl
4-[(2R,3R,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4-
-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoate (177 mg, 0.284
mmol) and g.HCl-MeOH (4 ml) was stirred at room temperature for 2
days. The mixture was concentrated in vacuo. The residue was
purified on TLC [CHCl.sub.3/MeOH (15/1)], to give methyl
4-[(2R,3R,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4-
-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoate (140 mg, 85%)
as a colorless amorphous solid IR (KBr) 3338, 2949, 1712, 1623,
1604, 1533 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 2.27 (m,
1H), 2.79 (m, 1H), 3.53 (dd, J=10.5, 5.9 Hz, 1H), 3.63 (s, 3H),
3.88 (s, 3H), 4.21 (d, J=7.8 Hz, 1H), 4.31 (m, 2H), 4.43 (dd,
J=9.8, 4.4 Hz, 1H), 4.52 (t, J=4.6 Hz, 1H), 6.71 (s, 1H), 6.80 (m,
3H), 6.99 (t, J=7.3 Hz, 1H), 7.16 (s, 1H), 7.21 (s, 1H), 7.39 (d,
J=8.1 Hz, 1H), 7.91 (d, J=8.6 Hz, 2H), 8.16 (d, J=8.3 Hz, 1H); Ms
(ESI) m/z 584 (M.sup.++1), 586 (M.sup.++3).
[0921] To a solution of methyl
4-[(2R,3R,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4-
-isopropylidenedioxy-2-pyrrolidinyl]methoxybenzoate (511 mg, 0.819
mmol) in THF (9.8 ml), 0.25 N NaOH (9.8 ml) was added. After
stirring at room temperature for 20 h, the mixture was acidified
with 1 N HCl and extracted with CHCl.sub.3-MeOH (10/1). The
combined extracts were dried over Na.sub.2SO.sub.4 and concentrated
in vacuo to give 143 (504 mg, 100%) as a colorless amorphous solid.
MW 610.05 IR (KBr) 3330, 2983, 2937, 1711, 1689, 1604, 1533, 1252
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.26 (s, 3H), 1.32
(s, 3H), 3.40 (m, 1H), 3.60 and 3.61 (d, J=2.5 Hz, 3H, amide
isomers), 3.62 (m, 1H), 3.78 and 3.83 (s, 3H, amideisomers), 4.16
(m, 2H), 4.42-4.98 (m, 3H), 6.74-7.15 (m, 6H), 7.28 (t, J=7.3 Hz,
1H), 7.43 (d, J=8.1 Hz, 1H), 7.78-7.97 (m, 4H), 8.08 (d, J=8.3 Hz,
1H), 8.89 (s, 1H), 8.92 (s, 1H), 12.68 (br, 1H); MS (ESI) m/z 610
(M.sup.++1), 612 (M.sup.++3).
Example 136
4-[(2R,3R,4S)-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4--
dihydroxy-2-pyrrolidinyl]methoxybenzoic acid
[0922] ##STR1765##
[0923] To a solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-3,4-dihydroxy--
2-pyrrolidinylmethoxy]benzoate (63 mg, 0.108 mmol) in THF (0.80
ml), 0.25 N NaOH (0.80 ml) was added. After stirring at room
temperature for 3 days, the mixture was acidified with 1 N HCl and
extracted with CHCl.sub.3-MeOH (10/1). The combined extracts were
dried over Na.sub.2SO.sub.4 and concentrated in vacuo to give 144
(61 mg, 100%) as a colorless amorphous solid. MW 569.99 IR (KBr)
3338, 1687, 1604, 1533, 1255, 1169, 1036 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 3.59 (d, J=5.5 Hz, 2H), 3.61 (m, 1H), 3.66
(dd, J=10.0, 7.1 Hz, 1H), 3.80 (s, 3H), 4.00-4.33 (m, 5H), 5.10
(br, 1H), 6.74 (d, J=8.3 Hz, 1H), 6.87 (s, 1H), 7.03 (m, 3H), 7.28
(t, J=8.3 Hz, 1H), 7.43 (d, J=6.6 Hz, 1H), 7.87 (d, J=8.5 Hz, 2H),
7.95 (d, J=8.3 Hz, 1H), 8.09 (d, J=8.3 Hz, 1H), 8.32 (s, 1H), 8.89
(s, 1H), 8.93 (s, 1H); MS (ESI) m/z 570 (M.sup.++1), 572
(M.sup.++3); Anal. Calcd for
C.sub.28H.sub.28ClN.sub.3O.sub.8.1.4H.sub.2O: C, 57.19; H, 5.14; N,
7.15. Found: C, 57.52; H, 5.22; N, 6.76.
Example 137
4-[1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phenyl-2--
(S)-pyrrolidinyl methoxy]benzoic acid
[0924] ##STR1766##
[0925] To a stirred solution of benzyl N-Boc-pyrroglutarate (8.93
g, 28.0 mmol) in THF (100 ml) was added phenyllithium (1.0 M in
Et.sub.2O-cyclohexane, 33.5 ml, 33.5 mmol) at -78.degree. C., and
the resulting mixture was gradually warmed up to -40.degree. C.,
then stirred overnight. aq.NH.sub.4Cl was added to the reaction
mixture, THF was removed in vacuo, then extracted with EtOAc. The
organic layer was washed with water and, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-EtOAc (4:1) as eluent,
then recrystallized from hexane-EtOAc to give benzyl
[2-(S)--N-Boc-amino)-5-oxo-6-phenyl]pentanoate (5.02 g, 45%) as a
colorless needles. mp 85-87.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.43 (s, 9H), 2.07-2.19 (m, 1H), 2.27-2.36 (m, 1H),
2.97-3.13 (m, 2H), 4.44 (brs, 1H), 5.19 (dd, J=25.2, 12.0 Hz, 2H),
5.19 (overlap, 1H), 7.28-7.98 (series of m, 10H); MS (ESI) m/z, 322
(M.sup.++H).
[0926] To a stirred solution of benzyl
[2-(S)--N-Boc-amino)-5-oxo-6-phenyl]pentanoate (2.20 g, 5.54 mmol)
in CH.sub.2Cl.sub.2 (50 ml) was added trifluoroacetic acid (15 ml)
at rt, and the resulting mixture was stirred for 2 h. The mixture
was concentrated in vacuo and poured into aq.NaHCO.sub.3, then
extracted with EtOAc. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo to give benzyl
5-phenyl-5-pyrroline-2-(S)-carboxylate (1.60 g, quant.) as
yellowish solid. The product was used for next reaction without
further purification: .sup.1H-NMR (CDCl.sub.3) .delta. 2.20-2.29
(m, 1H), 2.32-2.42 (m, 1H), 2.96-3.05 (m, 1H), 3.12-3.21 (m, 1H),
4.96-5.00 (m, 1H), 5.24 (s, 2H), 7.31-7.49 (m, 8H), 7.88-7.91 (m,
2H); MS (ESI) m/z, 280 (M.sup.++H).
[0927] A mixture of benzyl 5-phenyl-5-pyrroline-2-(S)-carboxylate
(1.59 g, 5.69 mmol) and Pd/C (10%, 128 mg) in MeOH (30 ml) was
stirred under H.sub.2 at rt for 28 h. The mixture was filtered and
the filtrate was concentrated in vacuo. The residue was dissolved
in CH.sub.3CN--H.sub.2O (3:2, 25 ml), then was added di-tert-butyl
dicarbonate (1.86 g, 8.54 mmol) and 1.0 M-NaOH (8.54 ml, 8.54
mmol), and the resulting mixture was stirred for 30 min. The
mixture was concentrated in vacuo and poured into aq.NaHCO.sub.3,
then extracted with EtOAc. The organic layer was washed with water,
saturated brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (9:1) and recrystallized from hexane-EtOAc
to give N-Boc-5-(R)-phenyl-(S)-proline (810 mg, 49%) as a colorless
solid. Mp 113-117.degree. C.; .sup.1H-NMR (CDCl.sub.3) .delta. 1.13
(s, 9H), 1.43 (brs, 1H), 1.96 (brs, 1H), 2.09 (brs, 1H), 2.31-2.34
(m, 1H), 2.46 (brs, 1H), 4.52 (brs, 1H), 4.69 (brs, 1H), 7.22-7.37
(m, 5H).
[0928] To a stirred solution of N-Boc-5-(R)-phenyl-2-(S)-proline
(1.14 g, 3.91 mmol) in THF (20 ml) was added 10M-BH.sub.3.Me.sub.2S
(780 ml, 7.82 mmol) at rt, and the resulting mixture was heated
under reflux for 30 min. The mixture was poured into aq. 1N-HCl and
extracted with EtOAc. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give N-Boc-2-(S)-hydroxymethyl-5-(R)-phenylpyrrolidine (1.11 g,
quant.) as a colorless oil: .sup.1H-NMR (CDCl.sub.3) .delta. 1.19
(brs, 9H), 1.65 (brs, 1H), 1.83-1.90 (m, 1H), 1.98-2.06 (m, 1H),
2.22-2.31 (m, 1H), 3.75-3.86 (m, 2H), 4.16-4.19 (m, 1H), 4.83 (t,
J=6.8 Hz, 1H), 4.89 (brs, 1H), 7.19-7.31 (m, 5H); MS (ESI) m/z, 278
(M.sup.++H).
[0929] To a stirred solution of
N-Boc-2-(S)-hydroxymethyl-5-(R)-phenylpyrrolidine (1.10 g, 3.97
mmol), triphenylphosphine (1.25 g, 4.76 mmol) and methyl
4-hydroxybenzoate (724 mg, 4.76 mmol) was added diisopropyl
azodicarboxylate (955 ml, 4.76 mmol) at rt, and the resulting
mixture was stirred at 60.degree. C. for 45 min. The mixture was
concentrated in vacuo, and the residue was chromatographed on
silica gel with hexane-EtOAc (4:1) as eluent to give methyl
4-[N-Boc-5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (1.31 g,
80%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.19 and
1.47 (brs, total 9H), 2.09-2.15 (m, 3H), 2.33-2.37 (m, 1H), 3.94
(s, 3H), 4.30 (brs, 1H), 4.41 (brs, 2H), 4.77 (brs, 1H), 7.03 (d,
J=8.8 Hz, 2H), 7.24-7.36 (m, 5H), 8.03-8.06 (m, 2H); MS (ESI) m/z,
412 (M.sup.++H).
[0930] To a stirred solution of methyl
4-[N-Boc-5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (1.28 g,
3.11 mmol) in CH.sub.2Cl.sub.2 (30 ml) was added trifluoroacetic
acid (10 ml) at rt, and the resulting mixture was stirred for 45
min. The mixture was concentrated in vacuo and poured into
aq.NaHCO.sub.3, then extracted with CHCl.sub.3. The organic layer
was washed with water, drying over anhydrous Na.sub.2SO.sub.4, and
concentrated in vacuo to give methyl
4-[5-(R)-phenyl-2-(S)-pyrrolidinyl methoxy]benzoate (363 mg,
quant.) as yellowish oil. The product was used for next reactions
without further purification. .sup.1H-NMR (CDCl.sub.3) .delta.
1.71-1.83 (m, 2H), 2.03-2.10 (m, 1H), 2.15-2.24 (m, 1H), 3.68-3.74
(m, 1H), 3.89 (s, 3H), 4.01-4.09 (m, 2H), 4.28 (t, J=7.2 Hz, 1H),
6.95 (d, J=8.8 Hz, 2H), 7.22-7.27 (m, 1H), 7.33 (t, J=8.0 Hz, 2H),
7.42 (d, J=7.6 Hz, 2H), 8.00 (d, J=8.8 Hz, 2H); MS (ESI) m/z, 312
(M.sup.++H) 353 (M.sup.++CH.sub.3CN).
[0931] To a stirred solution of methyl
4-[5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (135 mg, 0.43
mmol), 4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetic acid
(136 mg, 0.43 mmol) and N,N-dimethylaminopyridine (52.9 mg, 0.43
mmol) in DMF (10 ml) was added EDC.HCl (90.8 mg, 0.48 mmol) at rt,
and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with EtOAc. The organic
layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-EtOAc (1:5) as eluent to
give methyl 4-[1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenyl
acetyl]-5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (271 mg,
quant.) as a colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 2.00-2.18 (m, 3H), 2.31-2.41 (m, 1H), 2.27 (s, 3H), 3.31
(s, 2H), 3.67 (s, 3H), 3.89 (s, 3H), 4.35-4.48 (m, 2H), 4.60 (brs,
1H), 4.92 (t, J=6.8 Hz, 1H), 6.51 (d, J=8.4 Hz, 1H), 6.62 (s, 1H),
6.96 (d, J=8.8 Hz, 1H), 7.11-7.40 (series of m, 8H), 7.51 (d, J=8.0
Hz, 1H), 7.97-8.00 (m, 2H); MS (ESI) m/z, 608 (M.sup.++H).
[0932] To a stirred solution of methyl
4-[1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phenyl-2-
-(S)-pyrrolidinylmethoxy]benzoate (243 mg, 0.40 mmol) in MeOH-THF
(1:1, 10 ml) was added 1.0M-NaOH (2.4 ml, 2.40 mmol) at rt, and the
resulting mixture was heated at 60.degree. C. with stirring for 1.5
h. The reaction mixture was poured into 1N-HCl, then extracted with
CHCl.sub.3. The organic layer was washed with brine and dried over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) to
give 145 (224 mg, 94%) as a colorless amorphous solid. MW 593.67
.sup.1H-NMR (CD.sub.3OD), mixture of rotamars, .delta. 2.00-2.19
(m, 3H), 2.28 and 2.30 (s, total 3H), 2.45-2.49 (m, 1H), 3.37 (dd,
J=39, 16 Hz, 2H), 3.77 and 3.80 (s, total 3H), 3.92-5.18 (series of
m, 4H), 6.48-8.03 (series of m, 16H); MS (FAB) m/z, 594
(M.sup.++H).
Example 138
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phenyl-2--
(S)-pyrrolidinyl methoxy]benzoic acid
[0933] ##STR1767##
[0934] To a stirred solution of methyl
4-[5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (142 mg, 0.46
mmol), 4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid
(153 mg, 0.46 mmol) and N,N-dimethylaminopyridine (55.7 mg, 0.46
mmol) in DMF (10 ml) was added EDC.HCl (95.7 mg, 0.50 mmol) at rt,
and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with EtOAc. The organic
layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-EtOAc (1:5) as eluent to
give methyl 4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (260 mg,
90%) as a colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 2.00-2.19 (m, 3H), 2.35-2.44 (m, 1H), 3.35 (s, 2H), 3.76
(s, 3H), 3.89 (s, 3H), 4.38-4.48 (m, 2H), 4.63 (brs, 1H), 4.94 (t,
J=7.2 Hz, 1H), 6.53 (d, J=8.4 Hz, 1H), 6.66 (s, 1H), 6.96-7.01 (m,
3H), 7.12-7.42 (series of m, 8H), 7.87 (d, J=8.0 Hz, 1H), 7.98 (d,
J=8.8 Hz, 2H), 8.17-8.19 (m, 1H); MS (ESI) m/z, 627 (M.sup.+), 628
(M.sup.++H).
[0935] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phenyl-2-
-(S)-pyrrolidinylmethoxy]benzoate (251 mg, 0.40 mmol) in MeOH-THF
(1:1, 10 ml) was added 1.0M-NaOH (2.4 ml, 2.40 mmol) at rt, and the
resulting mixture was heated at 60.degree. C. with stirring for 1.5
h. The reaction mixture was poured into 1N-HCl, then extracted with
CHCl.sub.3. The organic layer was washed with brine and dried over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) to
give 146 (181 mg, 74%) as a colorless amorphous solid. MW 614.09
.sup.1H-NMR (CD.sub.3OD), mixture of rotamars, .delta. 1.99-2.19
(m, 3H), 2.42-2.53 (m, 1H), 3.38 (dd, J=39, 15 Hz, 2H), 3.79 and
3.80 (s, total 3H), 3.94-5.19 (series of m, 4H), 6.49-8.05 (series
of m, 16H); MS (FAB) m/z, 614 (M.sup.++H); Anal. Calcd for
C.sub.35H.sub.34ClN.sub.3O.sub.6.H.sub.2O: C, 65.06; H, 5.62; N,
6.50. Found: C, 65.03; H, 5.75; N, 6.45.
Example 139
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phenyl-2-(-
S)-pyrrolidinyl ethoxy]benzoic acid
[0936] ##STR1768##
[0937] To a stirred solution of methyl
4-[5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (146 mg, 0.47
mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (178
mg, 0.47 mmol) and N,N-dimethylaminopyridine (57.4 mg, 0.47 mmol)
in DMF (10 ml) was added EDC.HCl (99.0 mg, 0.52 mmol) at rt, and
the resulting mixture was stirred overnight. The reaction mixture
was poured into water and extracted with EtOAc. The organic layer
was washed with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with hexane-EtOAc (1:5) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (288 mg,
91%) as a colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 2.00-2.20 (m, 3H), 2.34-2.43 (m, 1H), 3.35 (s, 2H), 3.72
(s, 3H), 3.89 (s, 3H), 4.38-4.49 (m, 2H), 4.62 (brs, 1H), 4.94 (t,
J=7.2 Hz, 1H), 6.54 (d, J=8.4 Hz, 1H), 6.67 (s, 1H), 6.91-7.05 (m,
4H), 7.28-7.42 (series of m, 7H), 7.51 (d, J=8.0 Hz, 2H), 7.87 (d,
J=8.4 Hz, 1H), 7.99 (d, J=8.8 Hz, 2H), 8.14 (d, J=8.4 Hz, 2H); MS
(ESI) m/z, 672 (M.sup.+), 674 (M.sup.++2).
[0938] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phenyl-2--
(S)-pyrrolidinylmethoxy]benzoate (270 mg, 0.40 mmol) in MeOH-THF
(1:1, 10 ml) was added 1.0M-NaOH (2.0 ml, 2.0 mmol) at rt, and the
resulting mixture was heated at 60.degree. C. with stirring for 1
h. The reaction mixture was poured into 1N-HCl, then extracted with
CHCl.sub.3. The organic layer was washed with brine and dried over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) to
give 147 (212 mg, 80%) as a colorless amorphous solid. MW 658.54
.sup.1H-NMR (CD.sub.3OD), mixture of rotamars, .delta. 1.99-2.19
(m, 3H), 2.42-2.53 (m, 1H), 3.38 (dd, J=39, 16 Hz, 2H), 3.79 and
3.80 (s, total 3H), 3.94-5.19 (series of m, 4H), 6.49-8.00 (series
of m, 16H); MS (FAB) m/z, 658 (M.sup.+), 660 (M.sup.++2).
Example 140
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-pheny-
l-2-(S)-pyrrolidinyl methoxy]benzoic acid
[0939] ##STR1769##
[0940] To a stirred solution of methyl
4-[5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (109 mg, 0.35
mmol), 4-[N'-(2,4-dichlorophenyl)ureido]-3-methoxyphenylacetic acid
(129 mg, 0.35 mmol) and N,N-dimethylaminopyridine (42.8 mg, 0.35
mmol) in DMF (10 ml) was added EDC.HCl (73.4 mg, 0.39 mmol) at rt,
and the resulting mixture was stirred for 6 h The reaction mixture
was poured into water and extracted with EtOAc. The organic layer
was washed with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with hexane-EtOAc (1:4) as eluent to give methyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phen-
yl-2-(S)-pyrrolidinylmethoxy]benzoate (208 mg, 90%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 2.00-2.21 (m,
3H), 2.33-2.39 (m, 1H), 3.31 (s, 2H), 3.69 (s, 3H), 3.88 (s, 3H),
4.34-4.45 (m, 2H), 4.59 (brs, 1H), 4.93 (t, J=6.8 Hz, 1H), 6.47 (d,
J=8.0 Hz, 1H), 6.63 (s, 1H), 6.68 (s, 1H), 6.92-6.95 (m, 2H),
7.12-7.41 (series of n, 9H), 7.96-8.01 (m, 4H); MS (FAB) m/z, 662
(M.sup.++H).
[0941] To a stirred solution of methyl
4-[1-[4-[N'-(2,6-dichlorophenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-phen-
yl-2-(S)-pyrrolidinylmethoxy]benzoate (186 mg, 0.28 mmol) in
MeOH-THF (1:1, 10 ml) was added 1.0M-NaOH (1.4 ml, 1.4 mmol) at rt,
and the resulting mixture was heated at 60.degree. C. with stirring
for 2.5 h. The reaction mixture was poured into 1N-HCl, then
extracted with CHCl.sub.3. The organic layer was washed with brine
and dried over anhydrous Na.sub.2SO.sub.4, then concentrated in
vacuo. The residue was chromatographed on silica gel with
CHCl.sub.3-MeOH (10:1) to give 148 (166 mg, 91%) as a colorless
amorphous solid. MW 648.53 .sup.1H-NMR (CDCl.sub.3) .delta.
2.00-2.18 (m, 3H), 2.34-2.40 (m, 1H), 3.33 (s, 2H), 3.68 (s, 3H),
4.37-4.47 (m, 2H), 4.61 (brs, 1H), 4.94 (t, J=6.8 Hz, 1H), 6.48 (d,
J=8.0 Hz, 1H), 6.63 (s, 1H), 6.96 (d, J=8.4 Hz, 3H), 7.12-7.38
(series of m, 9H), 7.95 (d, J=8.0 Hz, 1H), 8.01 (d, J=8.8 Hz, 2H);
MS (FAB) m/z, 648 (M.sup.++H).
Example 141
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetyl]-5-(R)-phenyl-2-(S-
)-pyrrolidinyl methoxy]benzoic acid
[0942] ##STR1770##
[0943] To a stirred solution of methyl
4-[5-(R)-phenyl-2-(S)-pyrrolidinylmethoxy]benzoate (125 mg, 0.40
mmol), 4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetic acid (146
mg, 0.40 mmol) and N,N-dimethylaminopyridine (49.0 mg, 0.40 mmol)
in DMF (10 ml) was added EDC.HCl (84.1 mg, 0.44 mmol) at rt, and
the resulting mixture was stirred for 6 h. The reaction mixture was
poured into water and extracted with EtOAc. The organic layer was
washed with brine, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with hexane-EtOAc (1:4) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetyl]-5-(R)-phenyl-2-(-
S)-pyrrolidinylmethoxy]benzoate (238 mg, 90%) as a colorless
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.92 (s, 3H),
2.09-2.27 (m, 3H), 2.42-2.50 (m, 1H), 3.22-3.41 (m, 2H), 3.88 (s,
3H), 4.39 (d, J=4.4 Hz, 1H), 4.64 (brs, 1H), 5.00 (t, J=6.8 Hz,
1H), 6.72 (s, 1H), 6.81-6.93 (series of m, 8H), 7.22-7.42 (series
of m, 6H), 8.01 (d, J=8.4 Hz, 2H), 8.13 (d, J=8.0 Hz, 1H); MS (FAB)
m/z, 656 (M.sup.+), 658 (M.sup.++2).
[0944] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methylphenylacetyl]-5-(R)-phenyl-2-(-
S)-pyrrolidinylmethoxy]benzoate (216 mg, 0.33 mmol) in MeOH-THF
(1:1, 10 ml) was added 1.0M-NaOH (1.7 ml, 1.7 mmol) at rt, and the
resulting mixture was heated at 60.degree. C. with stirring for 2.5
h. The reaction mixture was poured into 1N-HCl, then extracted with
CHCl.sub.3. The organic layer was washed with brine and dried over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) to
give 149 (166 mg, 91%) as a colorless amorphous solid. MW 642.54
.sup.1H-NMR (CDCl.sub.3) .delta. 2.01 (s, 3H), 2.05-2.25 (m, 3H),
2.43-2.48 (m, 1H), 3.34 (dd, J=45, 16 Hz, 2H), 4.38-4.45 (m, 2H),
4.66 (brs, 1H), 4.99 (t, J=6.8 Hz, 1H), 6.77 (s, 1H), 6.82-6.88 (m,
2H), 6.94 (d, J=8.8 Hz, 2H), 7.15-7.55 (series of m, 10H), 8.00 (d,
J=8.8 Hz, 2H), 8.14 (d, J=7.2 Hz, 1H); MS (FAB) m/z 642 (M.sup.+),
644 (M.sup.++2).
Example 142
4-[1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-methyl-2--
(S)-pyrrolidinyl methoxy]benzoic acid
[0945] ##STR1771##
[0946] To a stirred solution of benzyl N-Boc-pyrroglutarate (7.55
g, 23.6 mmol) in THF (100 ml) was added MeLi (1.1 M in Et.sub.2O,
28.4 ml, 32.4 mmol) at -78.degree. C., and the resulting mixture
was gradually warned up to rt, then stirred overnight.
aq.NH.sub.4Cl was added to the reaction mixture, THF was removed in
vacuo, then extracted with EtOAc. The organic layer was washed with
water and, drying over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with hexane-EtOAc (3:1) as eluent to give benzyl
[2-(S)--(N-Boc-amino)-5-oxo-6-methyl]pentanoate (5.02 g, 45%) as a
colorless needles. mp 85-87.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.43 (s, 9H), 1.61-2.15 (series of m, 3H), 2.09 (s, 3H),
2.41-2.55 (m, 2H), 4.30 (brs, 1H), 4.70 (d, J=5.6 Hz, 1H),
5.12-5.21 (m, 2H), 7.29-7.37 (m, 5H); MS (ESI) m/z, 336
(M.sup.++H).
[0947] To a stirred solution of benzyl [2
(S)--N-Boc-amino)-5-oxo-6-methyl]pentanoate (4.46 g, 13.3 mmol) in
CH.sub.2Cl.sub.2 (50 ml) was added trifluoroacetic acid (20 ml) at
rt, and the resulting mixture was stirred for 1.5 h. The mixture
was concentrated in vacuo, and dissolved in toluene, then
evaporated to give benzyl 5-methyl-5-pyrroline-2-(S)-carboxylate
trifluoroacetic acid salt (5.74 g, quant.) as a crude brown oil.
This compound (1.97 g, 5.94 mmol) in MeOH (30 ml) was added Pd/C
(10%, 153 mg), and the resulting mixture was stirred for 3 days
under H.sub.2 atmosphere. The mixture was filtered, and the
filtrate was concentrated in vacuo to give
5-methyl-5-pyrrolidine-2-(S)-carboxylic acid trifluoroacetic acid
salt (956 mg, 66%) as a crude white solid. To a solution of this
compound (939 mg, 3.86 mmol) and di-tert-butyl dicarbonate in
MeCN-water (15:1, 16 ml) was added 1.0M-NaOH (8.49 mmol, 8.49 ml)
at rt, and the resulting mixture was stirred for 1 h. The resulting
mixture was evaporated and poured into aq.-1N-HCl, then extracted
with CHCl.sub.3/MeOH (5:1). The organic layer was dried over
anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH (7:1) to
give N-Boc-5-(R)-methyl-(S)-proline (711 mg, 80%) as a colorless
oil. .sup.1H-NMR (CD.sub.3OD) .delta. 1.27 (d, J=6.0 Hz, 3H),
1.41-1.46 (m, 9H), 1.62-1.64 (m, 1H), 1.96-2.01 (m, 2H), 2.22 (brs,
1H), 3.94 (brs, 1H), 4.17 (brs, 1H); MS (ESI) m/z, 230
(M.sup.++H).
[0948] To a stirred solution of N-Boc-5-(R)-methyl-2-(S)-proline
(1.03 g, 4.49 mmol) in THF (20 ml) was added 10M-BH.sub.3.Me.sub.2S
(1.57 ml, 15.7 mmol) at rt, and the resulting mixture was heated
under reflux for 5 h. The mixture was poured into aq. 1N-HCl and
extracted with EtOAc. The organic layer was dried over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with hexane-AcOEt (1:3) as eluent to
give N-Boc-2-(S)-hydroxymethyl-5-(R)-methylpyrrolidine (838 mg,
87%) as a colorless oil: .sup.1H-NMR (CDCl.sub.3) .delta. 1.17 (d,
J=6.0 Hz, 3H), 1.48 (s, 9H), 1.48-1.64 (m, 2H), 1.90-2.11 (m, 2H),
3.52-3.57 (m, 1H), 3.68-3.70 (m, 1H), 3.94-4.13 (m, 1H).
[0949] To a stirred solution of
N-Boc-2-(S)-hydroxymethyl-5-(R)-methylpyrrolidine (820 mg, 3.81
mmol), triphenylphosphine (1.10 g, 4.19 mmol) and methyl
4-hydroxybenzoate (580 mg, 3.81 mmol) was added diisopropyl
azodicarboxylate (841 ml, 4.19 mmol) at rt, and the resulting
mixture was stirred at 60.degree. C. for 1 h. The mixture was
concentrated in vacuo, and the residue was chromatographed on
silica gel with hexane-EtOAc (5:1) as eluent to give methyl
4-[N-Boc-5-(R)-methyl-2-(S)-pyrrolidinylmethoxy]benzoate (1.32 g,
80%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.24
(brs, 3H), 1.49 (s, 9H), 1.55-1.70 (m, 2H), 1.94-2.11 (m, 2H), 3.88
(s, 3H), 3.88 (overlap, 1H), 4.06-4.20 (m, 2H), 6.93-6.96 (m, 2H),
7.97 (d, J=8.8 Hz, 2H); MS (ESI) m/z, 350 (M.sup.++H).
[0950] To a stirred solution of methyl
4-[N-Boc-5-(R)-methyl-2-(S)-pyrrolidinylmethoxy]benzoate (1.29 g,
3.70 mmol) in CH.sub.2Cl.sub.2 (30 ml) was added trifluoroacetic
acid (10 ml) at rt, and the resulting mixture was stirred for 35
min. The mixture was concentrated in vacuo and poured into
aq.NaHCO.sub.3, then extracted with CHCl.sub.3. The organic layer
was washed with water, drying over anhydrous Na.sub.2SO.sub.4, and
concentrated in vacuo to give methyl
4-[5-(R)-methyl-2-(S)-pyrrolidinyl methoxy]benzoate (871 mg, 95%)
as a colorless oil. The product was used for next reactions without
further purification. .sup.1H-NMR (CDCl.sub.3) .delta. 1.18 (d,
J=6.4 Hz, 3H), 1.30-1.40 (m, 1H), 1.59-1.67 (m, 1H), 1.87-1.97 (m,
2H), 3.19-3.27 (m, 1H), 3.49-3.55 (m, 1H), 3.87 (s, 3H), 3.89-4.05
(m, 2H), 6.89 (d, J=8.8 Hz, 2H), 7.96 (d, J=8.8 Hz, 2H); MS (ESI)
m/z, 250 (M.sup.++H)
[0951] To a stirred solution of methyl
4-[5-(R)-methyl-2-(S)-pyrrolidinylmethoxy]benzoate (141 mg, 0.57
mmol), 4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetic acid
(178 mg, 0.57 mmol) and N,N-dimethylaminopyridine (69.0 mg, 0.57
mmol) in DMF (10 ml) was added EDC.HCl (120 mg, 0.62 mmol) at rt,
and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with EtOAc. The organic
layer was washed with brine, drying over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with EtOAc as eluent to give methyl
4-[1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenyl
acetyl]-5-(R)-methyl-2-(S)-pyrrolidinylmethoxy]benzoate (297 mg,
96%) as a colorless amorphous solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.24-1.34 (m, 3H), 1.93-2.18 (series of m, 4H), 2.28 (s,
3H), 3.65 (s, 3H), 3.88 (s, 3H), 3.62-3.87 (m, 3H), 4.11-4.38
(series of m, 3H), 6.42-8.06 (series of m, 13H); MS (ESI) m/z, 546
(M.sup.++H).
[0952] To a stirred solution of methyl
4-[1-[4-[N'-(2-methylphenyl)ureido]-3-methoxyphenylacetyl]-5-(R)-methyl-2-
-(S)-pyrrolidinylmethoxy]benzoate (279 mg, 0.51 mmol) in MeOH-THF
(1:1, 10 ml) was added 1.0M-NaOH (2.56 ml, 2.56 mmol) at rt, and
the resulting mixture was heated at 60.degree. C. with stirring for
2 h. The reaction mixture was poured into 1N-HCl, then extracted
with CHCl.sub.3. The organic layer was washed with brine and dried
over anhydrous Na.sub.2SO.sub.4, then concentrated in vacuo. The
residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(15:1) to give 150 (269 mg, 99%) as a colorless amorphous solid. MW
531.60 .sup.1H-NMR (CD.sub.3OD), mixture of rotamars, .delta.
1.28-1.35 (m, 3H), 1.74-2.21 (series of m, 4H), 2.28 (s, 3H),
3.71-4.37 (series of m, 6H), 6.76-7.99 (series of m, 11H); MS (ESI)
m/z, 532 (M.sup.++H).
Example 143
4-[trans-4-amino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(-
2S)-pyrrolidinyl]methoxybenzoic acid
[0953] ##STR1772##
[0954] To a solution of methyl
4-trans-4-amino-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)
methoxybenzoate (1.0 g, 2.86 mmol) and TEA (1.2 ml, 8.6 mmol) in
CH.sub.2Cl.sub.2 (20.0 ml) was added trifluoroacetic anhydride (720
mg, 3.43 mmol) at 0.degree. C. After stirred for 2.5 hr at room
temperature, water was added to the solution and extracted with
CH.sub.2Cl.sub.2. The extract was washed with water, then dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with EtOAc-n-hexane
(1:3, v/v) as eluent to give methyl
4-(trans-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl)met-
hoxybenzoate (940 mg, 74%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.46 (s, 9H), 2.02-2.18 (m, 1H), 2.41-2.52 (m,
1H), 3.30-3.45 (m, 1H), 3.80-3.90 (m, 1H), 3.88 (s, 3H), 4.00-4.30
(m, 3H), 4.65-4.75 (m, 1H), 6.50 (br s, 1H), 6.91-6.94 (m, 2H),
7.96-7.99 (m, 2H).
[0955] To a stirred solution of methyl
4-trans-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl)meth-
oxybenzoate (470 mg, 1.05 mmol) in CH.sub.2Cl.sub.2 (10.0 ml) was
added TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product,
3-methoxy-4-[N'-(2-methylphenyl)uredio]phenylacetic acid (314 mg,
1.0 mmol), HOBt (162 mg, 1.2 mmol), and triethylamine (417 ml, 3.0
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (288
mg, 1.5 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The organic layer
was washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with EtOAc-n-hexane (3:1, v/v) as eluent to give methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-triflu-
oroacetamido-(2S)-pyrrolidinyl]methoxybenzoate (350 mg, 52%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.01-2.10 (m, 1H),
2.31 (s, 3H), 2.42-2.48 (m, 1H), 3.45-3.50 (m, 1H), 3.56-3.59 (m,
5H), 3.89 (s, 3H), 4.07-4.14 (m, 2H), 4.38-4.42 (m, 1H), 4.50-4.60
(m, 1H), 4.72-4.80 (m, 1H), 6.33 (s, 1H), 6.60-6.85 (m, 3H),
7.06-7.26 (m, 3H), 7.48-7.52 (m, 1H), 7.93-8.05 (m, 3H).
##STR1773##
[0956] To a solution of methyl
4-trans-4-amino-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxybenzoate
(1.0 g, 2.86 mmol) and TEA (1.2 ml, 8.6 mmol) in CH.sub.2Cl.sub.2
(20.0 ml) was added trifluoroacetic anhydride (720 mg, 3.43 mmol)
at 0.degree. C. After stirred for 2.5 hr at room temperature, water
was added to the solution and extracted with CH.sub.2Cl.sub.2. The
extract was washed with water, then dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. The residue was purified by column
chromatography on silica gel with EtOAc-n-hexane (1:3, v/v) as
eluent to give methyl
4-(trans-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl)met-
hoxybenzoate (940 mg, 74%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.46 (s, 9H), 2.02-2.18 (m, 1H), 2.41-2.52 (m,
1H), 3.30-3.45 (m, 1H), 3.80-3.90 (m, 1H), 3.88 (s, 3H), 4.00-4.30
(m, 3H), 4.65-4.75 (m, if), 6.50 (br s, 1H), 6.91-6.94 (m, 2H),
7.96-7.99 (m, 2H).
[0957] To a stirred solution of methyl
4-trans-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl)meth-
oxybenzoate (470 mg, 1.05 mmol) in CH.sub.2Cl.sub.2 (10.0 ml) was
added TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product,
3-methoxy-4-[1N'-(2-methylphenyl)uredio]phenylacetic acid (314 mg,
1.0 mmol), HOBt (162 mg, 1.2 mmol), and triethylamine (417 ml, 3.0
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (288
mg, 1.5 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The organic layer
was washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with EtOAc-n-hexane (3:1, v/v) as eluent to give methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-triflu-
oroacetamido-(2S)-pyrrolidinyl]methoxybenzoate (350 mg, 52%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.01-2.10 (m, 1H),
2.31 (s, 3H), 2.42-2.48 (m, 1H), 3.45-3.50 (m, 1H), 3.56-3.59 (m,
5H), 3.89 (s, 3H), 4.07-4.14 (m, 2H), 4.38-4.42 (m, 1H), 4.50-4.60
(m, 1H), 4.72-4.80 (m, 1H), 6.33 (s, 3H), 6.60-6.85 (m, 3H),
7.06-7.26 (m, 3H), 7.48-7.52 (m, 1H), 7.93-8.05 (m, 3H).
[0958] To a stirred solution of methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-triflu-
oroacetamido-(2S)-pyrrolidinyl]methoxybenzoate (150 mg, 0.23 mmol)
in THF (3.0 ml) and MeOH (2.0 ml) was added 1N NaOH (0.70 ml, 0.70
mmol). The mixture was stirred at 60.degree. C. for 18 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 151 (100 mg, 81%) as a white
crystalline solid. MW 532.59 mp 170-171.degree. C.; IR (KBr) 3264,
2937, 1604, 1535, 1415, 1376, 1255, 1224, 1033 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.80-1.90 (m, 1H), 2.10-2.20 (m,
1H), 2.24 (s, 3H), 3.55-3.80 (m, 3H), 3.57 (s, 2H), 4.08-4.18 (m,
2H), 4.36-4.60 (m, 1H), 6.72-7.16 (m, 7H), 7.77-8.01 (m, 4H), 8.46
(s, 1H), 8.54 (s, 1H); MS (FAB) m/z 532 (M.sup.++1); Anal. calcd
for C.sub.29H.sub.32N.sub.4O.sub.6.2.0H.sub.2O: C, 61.26; H, 6.38;
N, 9.85. Found: C, 61.07; H, 6.32; N, 9.58.
Example 144
methyl
4-[trans-4-amino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinyl]methoxybenzoate HCl salt
[0959] ##STR1774##
[0960] To a stirred solution of methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-triflu-
oroacetamido-(2S)-pyrrolidinyl]methoxybenzoate (200 mg, 0.31 mmol)
in MeOH (4.0 mmol) was added water (2.0 ml) and K.sub.2CO.sub.3
(138 mg, 1.0 mmol) at room temperature. After stirred for 18 hr at
room temperature, water was added to the mixture and extracted with
CH.sub.2Cl.sub.2. The extract was washed with water, then dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with
MeOH--CH.sub.2Cl.sub.2 (5:95 to 15:85, v/v) as eluent. The product
was dissolved in EtOH (5.0 ml), and 1N HCl (in EtOH) (1.0 ml, 1.0
mmol) was added thereto. The mixture was concentrated in vacuo to
give 152 (120 mg, 63%) as an amorphous solid. MW 546.61 IR (KBr)
3382, 2948, 2879, 1604, 1533, 1286, 1255, 771 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25 (s, 3H), 2.10-2.30 (m, 2H),
3.59-3.70 (m, 3H), 3.77-3.80 (m, 8H), 4.00-4.24 (m, 2H), 4.47-4.67
(m, 1H), 6.70-7.16 (m, 7H), 7.77-8.00 (m, 4H), 8.49 (s, 1H), 8.55
(s, 1H); MS (FAB) m/z 547 (M.sup.++1); Anal. calcd for
C.sub.30H.sub.34N.sub.4O.sub.6.HCl.1.4H.sub.2O: C, 59.24; H, 6.26;
N, 9.21; Cl, 5.83. Found: C, 59.42; H, 6.42; N, 9.04; Cl, 6.11.
Example 145
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-methyla-
mino-(2S)-pyrrolidinyl]methoxybenzoic acid
[0961] ##STR1775##
[0962] To a stirred solution of methyl
4-(trans-1-tert-butoxycarbonyl-4-trifluoroacetamido-(2S)-pyrrolidinyl)met-
hoxybenzoate (520 mg, 1.17 mmol) in DMF (10.0 ml) was added
K.sub.2CO.sub.3 (321 mg, 2.33 mmol) and MeI (330 mg, 2.33 mmol) at
room temperature. The reaction mixture was stirred at 50.degree. C.
for 18 hr. Water was added to the mixture and extracted with EtOAc.
The organic layer was washed with water, then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with EtOAc-n-hexane
(1:2, v/v) as eluent to give methyl
4-[trans-1-tert-butoxycarbonyl-4-(N-methyl-trifluoroacetamido)-(2S)-pyrro-
lidinyl]methoxybenzoate (390 mg, 73%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.46 (9H, s), 2.12-2.40 (m, 2H),
2.96 and 3.05 (each s, total 3H), 3.28-3.70 (m, 2H), 3.88 (s, 3H),
3.95-4.42 (m, 3H), 5.10-5.40 (m, 1H), 6.89-6.91 (m, 2H), 7.96-8.00
(m, 2H).
[0963] To a stirred solution of methyl
4-[trans-1-tert-butoxycarbonyl-4-(N-methyl-trifluoro
acetoamido)-(2S)-pyrrolidinyl]methoxybenzoate (390 mg, 0.85 mmol)
in CH.sub.2Cl.sub.2 (8.0 ml) was added TFA (5.0 ml) at 0.degree. C.
The reaction mixture was stirred at room temperature for 0.5 hr.
The mixture was concentrated in vacuo. Sat. NaHCO.sub.3 was added
to the residue, and extracted with CH.sub.2Cl.sub.2. The extract
was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The crude product was used to the subsequent
reaction without further purification. To a stirred solution of the
crude product, 3-methoxy-4-[N'-(2-methylphenyl)uredio]phenylacetic
acid (279 mg, 0.89 mmol), HOBt (143 mg, 1.1 mmol), and
triethylamine (246 ml, 1.77 mmol) in THF (8.0 ml) and MeCN (8.0 ml)
was added EDC.HCl (255 mg, 1.3 mmol) at 0.degree. C. The reaction
mixture was stirred at room temperature for 16 hr, and concentrated
in vacuo. Water was added to the residue, and extracted with EtOAc.
The extract was washed with sat. NaHCO.sub.3, 2-M citric acid, and
sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with EtOAc-n-hexane (4:1, v/v) as
eluent to give methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(N-met-
hyl-trifluoroacetoamido)-(2S)-pyrrolidinyl]methoxybenzoate (480 mg,
82%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.18-2.35
(m, 2H), 2.31 (s, 3H), 2.87 and 2.97 (each s, total 3H), 3.45-3.46
(m, 3H), 3.47 (s, 3H), 3.49 (s, 2H), 3.88 (s, 3H), 4.30-4.70 (m,
2H), 5.20-5.40 (m, 1H), 6.38-6.43 (m, 1H), 6.67-6.86 (m, 4H),
7.09-7.24 (m, 4H), 7.51-7.54 (m, 1H), 7.93-8.08 (m, 3H).
[0964] To a stirred solution of methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(N-met-
hyl-N-trifluoroacetylamino)-(2S)-pyrrolidinyl]methoxybenzoate (240
mg, 0.37 mmol) in THF (5.0 ml) and MeOH (3.0 ml) was added 1N NaOH
(1.27 ml, 1.27 mmol). The mixture was stirred at 60.degree. C. for
18 hr. The mixture was concentrated in vacuo, water was added
thereto, and neutralized with 1N HCl. The resulting solid was
collected, washed with water, and dried in vacuo to give 153 (140
mg, 70%) as a white crystalline solid. MW 546.61 mp 162-164.degree.
C.; IR (KBr) 3338, 1604, 1535, 1255, 1033, 755 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.85-1.95 (m, 1H), 2.10-2.20 (m,
1H), 2.24 (s, 3H), 2.34 and 2.39 (each s, total 3H), 3.41-3.71 (m,
3H), 3.58 (s, 2H), 3.80 (s, 3H), 4.05-4.20 (m, 2H), 4.36-4.60 (m,
1H), 6.73-7.16 (m, 7H), 7.77-8.01 (m, 4H), 8.45 (s, 1H), 8.53 (s,
1H); MS (FAB) m/z 547 (M.sup.++1); Anal. calcd for
C.sub.30H.sub.34N.sub.4O.sub.6.2.5H.sub.2O: C, 60.90; H, 6.64; N,
9.47. Found: C, 61.01; H, 6.50; N, 9.31.
Example 146
methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4--
methylamino-(2S)-pyrrolidinyl]methoxybenzoate
[0965] ##STR1776##
[0966] To a stirred solution of methyl
4-[trans-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-4-(N-met-
hyltrifluoroacetamido)-(2S)-pyrrolidinyl]methoxybenzoate (240 mg,
0.36 mmol) in THF (5.0 mmol) and MeOH (5.0 ml) was added water (2.0
ml) and K.sub.2CO.sub.3 (138 mg, 1.0 mmol) at room temperature.
After stirred for 18 hr at room temperature, water was added to the
mixture and extracted with CH.sub.2Cl.sub.2. The extract was washed
with water, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with MeOH--CH.sub.2Cl.sub.2 (5/95 to 20/80, v/v) as eluent. The
product was dissolved in EtOH (5.0 ml), and 1N HCl (in EtOH) (0.71
ml, 0.71 mmol) was added thereto. The mixture was concentrated in
vacuo to give 154 (180 mg, 85%) as an amorphous solid. MW 560.64 IR
(KBr) 3311, 2692, 2453, 1712, 1604, 1533 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.24 (s, 3H), 2.15-2.30 (m, 2H), 2.60 (br s,
3H), 3.60-4.20 (m, 5H), 3.78-3.81 (m, 8H), 4.47-4.70 (m, 1H),
6.71-7.16 (m, 7H), 7.77-8.00 (m, 4H), 8.48 (s, 1H), 8.55 (s, 1H),
9.21 (br s, 2H); MS (FAB) m/z 561 (M.sup.++1); Anal. calcd for
C.sub.31H.sub.36N.sub.4O.sub.6.HCl.1.4H.sub.2O: C, 59.83; H, 6.45;
N, 9.00; Cl, 5.70. Found: C, 60.08; H, 6.51; N, 8.68; Cl, 5.99.
Example 147
4-[trans-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyla-
cetyl]-(2S)-pyrrolidinyl]methoxybenzoic acid
[0967] ##STR1777##
[0968] To a stirred solution of
trans-1-tert-butoxycarbonyl-(2S)-hydroxymethyl-4-hydroxypyrrolidine
(2.17 g, 10.0 mmol) and imidazole (2.04 g, 30.0 mmol) in DMF (50
ml) was added TBDPS-Cl (3.03 g, 11.0 mmol) at 0.degree. C. The
reaction mixture was stirred at room temperature for 18 hr. Water
was added thereto, and extracted with EtOAc. The extract was washed
with water, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (3:2, v/v) as eluent to give
trans-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-4-hyd-
roxypyrrolidine (1.5 g, 33%) as a white crystalline solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.03 (s9H), 1.25 and 1.32 (each s,
9H), 1.90-2.10 (m, 1H), 2.30-2.40 (m, 1H), 3.40-3.80 (m, 3H),
3.95-4.15 (m, 2H), 4.45-4.55 (m, 1H), 7.37-7.39 (m, 6H), 7.63-7.64
(m, 4H).
[0969] To a stirred solution of
trans-1-tert-butoxycarbonyl-(2S)-tert-butyldiphenylsilyloxy)methyl-4-hydr-
oxypyrrolidine (910 mg, 2.0 mmol) and Ph.sub.3P (628 mg, 2.4 mmol)
in THF (20 ml) was added CBr.sub.4 (993 mg, 3.0 mmol) at room
temperature. The reaction mixture was stirred at room temperature
for 0.5 hr. n-Hexane (40 ml) was added thereto. The resulting solid
was filtered off, and dried in vacuo. The residue was purified by
column chromatography on silica gel with n-hexane to n-hexane-EtOAc
(3:2, v/v) as eluent to give
cis-4-bromo-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-
pyrrolidine (1.0 g, quant.) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.06 (s, 9H), 1.31 and 1.45 (each s, 9H), 2.63
(m, 2H), 3.49 (m, 1H), 3.89-4.14 (m, 5H), 7.35-7.42 (m, 6H),
7.64-7.66 (, 4Hm).
[0970] To a stirred solution of
cis-4-bromo-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-
pyrrolidine (480 mg, 0.93 mmol) in DMF (5 ml) was added NaN.sub.3
(241 mg, 3.70 mmol) at room temperature. The reaction mixture was
stirred at 70 for 3 days. Water was added thereto, and extracted
with EtOAc. The extract was washed with water, then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. The
solution of the crude residue in EtOH (10 ml) was hydrogenated over
10% Pd--C under an atmospheric pressure at room temperature for 4
hr. The catalyst was filtered off, and the filtrate was
concentrated in vacuo to give
trans-4-amino-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)meth-
ylpyrrolidine (400 mg, 95%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.06 (s, 9H), 1.32 and 1.45 (each s, total
9H), 2.20-2.35 (m, 1H), 3.05-3.18 (m, 1H), 3.55-4.05 (m, 6H),
7.35-7.41 (m, 6H), 7.61-7.69 (m, 4H).
[0971] To a stirred solution of
trans-4-amino-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)meth-
ylpyrrolidine (400 mg, 0.88 mmol), AcOH (120 ml, 2.0 mmol), and 37%
HCHO aq (500 ml) in MeOH (10 ml) was added NaBH.sub.3CN (111 mg,
1.76 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. After concentrated in vacuo, water was added
and extracted with CH.sub.2Cl.sub.2. The extract was dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with
MeOH--CH.sub.2Cl.sub.2 (3:97, v/v) as eluent to give
trans-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-4-met-
hylaminopyrrolidine (330 mg, 78%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.06 (s, 9H), 1.33 and 1.45 (each s, total
9H), 1.80-2.25 (m, 2H), 2.23 (br s, 6H), 2.95-4.05 (m, 6H),
7.36-7.39 (m, 6H), 7.63-7.65 (m, 4H).
[0972] To a stirred solution of
trans-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-4-dim-
ethylaminopyrrolidine (330 mg, 0.68 mmol) in THF (5 ml) was added
TBAF (1.0 M solution in THF, 1.0 ml, 1.0 mmol) at 0.degree. C. The
reaction mixture was stirred at room temperature for 2 hr. The
mixture was concentrated in vacuo. The residue was purified by
column chromatography on silica gel with MeOH--CH.sub.2Cl.sub.2
(3:97 to 20:80, v/v) as eluent to give
trans-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-hydroxymethylpyr-
rolidine (180 mg, quant) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.47 (s, 9H), 2.23 (s, 6H), 1.65-1.75 (m, 2H),
2.75-4.10 (m, 4H), 3.61 (d, J=5.6 Hz, 2H).
[0973] To a stirred solution of
trans-1-tert-butoxycarbonyl-4-dimethylamino(2S)-hydroxymethyl
pyrrolidine (180 mg, 0.73 mmol), methyl 4-hydroxybenzoate (114 mg,
0.75 mmol), and Ph.sub.3P (296 mg, 1.13 mmol) in THF (10 ml) was
added DIAD (227 mg, 1.13 mmol) at 0.degree. C. The reaction mixture
was stirred at 70.degree. C. for 18 hr. The mixture was
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (1:2, v/v) to
MeOH--CH.sub.2Cl.sub.2 (5:95, v/v) as eluent to give methyl
4-[trans-1-tert-butoxycarbonyl-4-dimethylamino(2S)-pyrrolidinyl]methoxybe-
nzoate (180 mg, 68%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.46 (s, 9H), 1.80-1.95 (m, 1H), 2.20-2.23 (m, 1H), 2.24
(s, 6H), 2.90-2.95 (m, 1H), 3.10-3.30 (m, 1H), 3.50-3.65 (m, 1H),
3.88 (s, 3H), 3.95-4.35 (m, 3H), 6.93-6.95 (m, 2H), 7.96-7.98 (m,
2H).
[0974] To a stirred solution of methyl
4-(trans-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl)methoxyb-
enzoate (200 mg, 0.53 mmol) in CH.sub.2Cl.sub.2 (6 ml) was added
TFA (3 ml) at 0.degree. C. The reaction mixture was stirred at room
temperature for 0.5 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product,
3-methoxy-4-[N'-(2-methylphenyl)uredio]phenylacetic acid (166 mg,
0.53 mmol), HOBt (71 mg, 0.53 mmol), and triethylamine (140 ml,
1.10 mmol) in THF (5 ml) and MeCN (5 ml) was added EDC.HCl (152 mg,
0.79 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with EtOAc to CH.sub.2Cl.sub.2-MeOH
(8:92, v/v) as eluent to give methyl
4-[trans-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl-
acetyl]-(2S)-pyrrolidinyl]methoxybenzoate (260 mg, 86%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.95-2.15 (m, 3H),
2.23 (s, 6H), 2.31 (s, 3H), 3.30-3.34 (m, 1H), 3.57 (s, 2H), 3.61
(s, 3H), 3.70-3.75 (m, 1H), 4.11-4.15 (m, 2H), 4.45-4.50 (m, 1H),
6.34 (s, 1H), 6.72-6.88 (m, 4H), 7.08-7.24 (m, 4H), 7.51-7.53 (m,
1H), 7.92-8.07 (m, 3H).
[0975] To a stirred solution of methyl
4-[trans-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl-
acetyl]-2-pyrrolidinyl]methoxybenzoate (260 mg, 0.45 mmol) in THF
(4.0 ml) and MeOH (2.0 ml) was added 1N NaOH (0.90 ml, 0.90 mmol).
The mixture was stirred at 70.degree. C. for 24 hr. The mixture was
concentrate in vacuo, water was added thereto, and neutralized with
1N HCl. The resulting solid was collected, washed with water, and
dried in vacuo to give 155 (200 mg, 79%) as a white crystalline
solid. MW 560.64 mp 145-150.degree. C.; IR (KBr) 3355, 2948, 1698,
1604, 1533, 1454, 1417, 1255, 1226, 1166, 1035, 755 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.82-1.98 (m, 1H), 2.08-2.11 (m,
1H), 2.20 (s, 6H), 2.25 (s, 3H), 3.40-3.60 (m, 3H), 3.64 (s, 2H),
3.82 (s, 3H), 4.01-4.16 (m, 2H), 4.36 (m, 1H), 6.74-7.15 (m, 7H),
7.77-8.02 (m, 4H), 8.44 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 561
(M.sup.++1); Anal. calcd for C.sub.31H.sub.36N.sub.4O.sub.6.1.2H:
C, 63.95; H, 6.65; N, 9.62. Found: C, 63.82; H, 6.72; N, 9.44.
Example 148
methyl
4-[trans-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]-
phenylacetyl]-(2S)-pyrrolidinyl]methoxybenzoate HCl salt
[0976] ##STR1778##
[0977] To a stirred solution of
trans-4-[4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl-
acetyl-(2S)-pyrrolidinyl]methoxybenzoic acid (80 mg, 0.14 mmol) in
toluene (4.0 ml) and MeOH (1.0 ml) was added TMSCHN.sub.2 (2.0 M in
hexane, 100 ml, 0.20 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 1.5 hr. The mixture was
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with MeOH--CH.sub.2Cl.sub.2 (5:95,
v/v) as eluent The product was dissolved in EtOH (5.0 ml), and 1N
HCl (in EtOH) (244 .mu.l, 0.244 mmol) was added thereto. The
mixture was concentrated in vacuo to give 156 (72 mg, 88%) as an
amorphous solid. MW 574.67 IR (KBr) 3345, 2950, 2586, 1712, 1604,
1511, 1454, 1284, 1255, 1170, 1114, 1029, 850, 771 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25 (s, 3H), 2.35-2.37 (m, 2H),
2.77-2.81 (m, 6H), 3.62-3.71 (m, 2H), 3.79-3.81 (m, 8H), 3.99-4.16
(m, 3H), 4.50-4.70 (m, 1H), 6.74-7.16 (m, 7H), 7.77-8.01 (m, 4H),
8.48 (s, 1H), 8.55 (s, 1H); Anal. calcd for
C.sub.32H.sub.38N.sub.4O.sub.6.1.0HCl 1.2H.sub.2O: C, 60.74; H,
6.59; N, 8.85. Found: C, 61.03; H, 6.78; N, 8.33.
Example 149
4-cis-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]-(2S)-pyrrolidinyl]methoxybenzoic acid
[0978] ##STR1779##
[0979] To a stirred solution of
cis-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-4-hydro-
xypyrrolidine (1.82 mg, 4.0 mmol), phthalimide (647 mg, 4.4 mmol),
and Ph.sub.3P (1.26 g, 4.8 mmol) in THF (20 ml) was added DIAD (889
mg, 4.4 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 18 hr. The mixture was concentrated in vacuo.
The residue was purified by column chromatography on silica gel
with n-hexane-EtOAc (5/1, v/v) as eluent to give
N-[cis-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-4-py-
rrolidinyl]phthalimide (1.6 g, 69%) as an amorphous solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.07 (s, 9H), 1.30 and 1.44 (each
s, total 9H), 2.27-2.37 (m, 1H), 2.94-2.96 (m, 1H), 3.81-4.09 (m,
5H), 4.72 (m, 1H), 7.37-7.38 (m, 6H), 7.67-7.74 (m, 6H), 7.84-7.86
(m, 2H).
[0980] To a stirred solution of
N-[cis-1-tert-butoxycarbonyl-(2S)-(tert-butyldiphenylsilyloxy)methyl-4-py-
rrolidinyl]phthalimide (1.60 g, 2.74 mmol) in EtOH (8 ml) was added
NH.sub.2NH.sub.2.H.sub.2O (206 mg, 4.11 mmol) at room temperature.
The reaction mixture was stirred at 70 for 1 hr. The mixture was
concentrated in vacuo. The resulting solid was filtered off, and
washed with CHCl.sub.3. The filtrate was concentrated in vacuo. The
resulting solid was filtered off, and washed with CHCl.sub.3. The
filtrate was concentrated in vacuo to give
cis-4-amino-1-tert-butoxycarbonyl-(2S)-tert-butyl
diphenylsilyloxy)methylpyrrolidine (1.3 g, quant) as a pale yellow
oil. The crude product was used to the subsequent reaction without
further purification. .sup.1H-NMR (CDCl.sub.3) .delta. 1.06 (s,
9H), 1.30 and 1.45 (each s, total 9H), 1.59 (m, 1H), 1.85 (m, 1H),
2.94 (m, 1H), 3.44 (m, 1H), 3.78-4.07 (m, 4H), 7.36-7.41 (m, 6H),
7.51-7.65 (m, 4H).
[0981] To a stirred solution of
cis-4-amino-1-tert-butoxycarbonyl-(2S)-tert-butyldiphenylilyloxy)methylpy-
rrolidine (1.24 g, 2.74 mmol), AcOH (374 .mu.l, 5.48 mmol), and 37%
HCHO aq (1.0 ml) in MeOH (20 ml) was added NaBH.sub.3CN (345 mg,
5.48 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. After concentrated in vacuo, water was added
and extracted with CH.sub.2Cl.sub.2. The extract was dried over
Na.sub.2SO.sub.4 and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with
MeOH--CH.sub.2Cl.sub.2 (3/97, v/v) as eluent to give
cis-1-tert-butoxycarbonyl-(2S)-tert-butyldiphenylsilyloxy)methyl-4-dimeth-
ylamino pyrrolidine (1.1 g, 83%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.05 (s, 9H), 1.29 and 1.45 (each s, total
9H), 1.95-2.04 (m, 1H), 2.20-2.26 (m, 1H), 2.27 (s, 6H), 2.54 (m,
1H), 3.00-3.02 (m, 1H), 3.62-4.03 (m, 4H), 7.34-7.41 (m, 6H),
7.63-7.65 (m, 4H).
[0982] To a stirred solution of
cis-1-tert-butoxycarbonyl-2-(tert-butyldiphenylsilyloxy)methyl-4-methyl
amino pyrrolidine (10.1 g, 2.27 mmol) in THF (10 ml) was added TBAF
(1.0 M solution in THF) (4.5 ml, 4.5 mmol) at 0.degree. C. The
reaction mixture was stirred at room temperature for 2 hr. The
mixture was concentrated in vacuo. The residue was purified by
column chromatography on silica gel with MeOH--CH.sub.2Cl.sub.2
(3/97 to 20/80, v/v) as eluent to give
cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-hydroxymethylpyrro-
lidine (580 mg, quant.) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.47 (s, 9H), 1.25-1.96 (m, 2H), 2.25 (s, 6H),
2.53-2.58 (m, 1H), 3.17-4.02 (m, 5H).
[0983] To a stirred solution of
cis-1-tert-butoxycarbonyl-4-methylamino-(2S)-hydroxymethylpyrrolidine
(555 mg, 2.27 mmol), methyl 4-hydroxybenzoate (380 mg, 2.5 mmol),
and Ph.sub.3P (1.07 g, 4.09 mmol) in THF (10 ml) was added DIAD
(826 mg, 4.09 mmol) at 0.degree. C. The reaction mixture was
stirred at 70.degree. C. for 18 hr. The mixture was concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1/2, v/v) MeOH--CH.sub.2Cl.sub.2 (5/95,
v/v) as eluent to give methyl
4-cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl)methoxy
benzoate (260 mg, 30%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.45 (s, 9H), 1.70-1.90 (m, 1H), 2.26 (s, 6H),
2.33 (m, 1H), 2.57 (m, 1H), 3.06 (m, 1H), 3.85-4.23 (m, 4H), 3.88
(s, 3H), 6.93 (m, 2H), 7.95 (m, 2H).
[0984] To a stirred solution of methyl
4-(cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl)methoxyben-
zoate (208 mg, 0.55 mmol) in CH.sub.2Cl.sub.2 (6 ml) was added TFA
(3 ml) at 0.degree. C. The reaction mixture was stirred at room
temperature for 0.5 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product,
3-methoxy-4-[N'-(2-methylphenyl)uredio]phenylacetic acid (173 mg,
0.55 mmol), HOBt (74 mg, 0.55 mmol), and triethylamine (153 .mu.l,
1.1 mmol) in THF (6 ml) and MeCN (6 ml) was added EDC.HCl (160 mg,
0.83 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with EtOAc--CH.sub.2Cl.sub.2-MeOH
(5/95, v/v) as eluent to give methyl 4-[cis-4
dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinyl]methoxybenzoate (270 mg, 47%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.95-2.04 (m, 1H),
2.25 (s, 6H), 2.32 (s, 3H), 2.61 (m, 1H), 3.21 (m, 1H), 3.56-3.58
(m, 5H), 3.80-3.83 (m, 1H), 3.88 (s, 3H), 4.18-4.20 (m, 1H),
4.41-4.45 (m, 2H), 6.36 (s, 1H), 6.68-6.85 (m, 4H), 7.08-7.25 (m,
4H), 7.52-7.55 (m, 1H), 7.91-8.07 (m, 3H).
[0985] To a stirred solution of methyl
4-[cis-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinyl]methoxybenzoate (270 mg, 0.47 mmol) in THF
(4.0 ml) and MeOH (2.0 ml) was added 1N NaOH (1.0 ml, 1.0 mmol).
The mixture was stirred at 70.degree. C. for 18 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 157 (170 mg, 65%) as a white crystalline
solid. MW 560.64 mp 147-150.degree. C.; IR (KBr) 3353, 2952, 1700,
1604, 1533, 1454, 1415, 1255, 1166, 1035, 755 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) .delta.: 1.83-1.84 (m, 1H), 2.08-2.10
(m, 1H), 2.21 (br s, 6H), 2.24 (s, 3H), 3.00 (m, 2H), 3.60 (s, 2H),
3.78 (s, 3H), 3.85-4.29 (m, 4H), 6.71-7.16 (m, 7H), 7.77-8.01 (m,
4Hm), 8.46 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 561 (M+H).sup.+;
Anal. calcd for C.sub.31H.sub.36N.sub.4O.sub.6.2H.sub.2O: C, 62.40;
H, 6.76; N, 9.39. Found: C, 62.51; H, 6.60; N, 9.36.
Example 150
methyl
4-[cis-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]ph-
enylacetyl]-(2S)-pyrrolidinyl]methoxybenzoate HCl salt
[0986] ##STR1780##
[0987] To a stirred solution of
4-[cis-4-dimethylamino-1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylac-
etyl]-(2S)-pyrrolidinyl]methoxybenzoic acid (80 mg, 0.14 mmol) in
toluene (4.0 ml) and MeOH (1.0 ml) was added TMSCHN.sub.2 (2.0 M in
hexane) (100 .mu.l, 0.20 mmol) at 0.degree. C. The reaction mixture
was stirred at room temperature for 1.5 hr. The mixture was
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with MeOH--CH.sub.2Cl.sub.2 (5/95,
v/v) as eluent. The product was dissolved in EtOH (5.0 ml), and 1N
HCl (in EtOH) (244 .mu.l, 0.244 mmol) was added thereto. The
mixture was concentrated in vacuo to give 158 (75 mg, 79%) as an
amorphous solid. MW 574.67 IR (KBr) 3345, 2950, 2456, 1712, 1646,
1604, 1511, 1454, 1434, 1415, 1284, 1257, 1168, 1114, 1031, 771
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) 2.10-2.20 (m, 2H), 2.25 (s,
3H), 2.83 (m, 6H), 3.60-3.62 (m, 2H), 3.76-3.81 (m, 8H), 4.20-4.33
(m, 4H), 6.71-7.17 (m, 6H), 7.77-7.98 (m, 5H), 8.47 (s, 1H), 8.55
(s, 1); MS (FAB) m/z 574 (M+H).sup.+; Anal. calcd for
C.sub.32H.sub.38N.sub.4O.sub.6.1.0HCl 1.3H.sub.2O: C, 60.57; H,
6.61; N, 8.83. Found: C, 60.80; H, 6.82; N, 8.44.
Example 151
4-[trans-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-dimethy-
lamino-(2S)-pyrrolidinyl]methoxybenzoic acid
[0988] ##STR1781##
[0989] To a stirred solution of methyl
4-(trans-1-tert-butoxycarbonyl-4-dimethylamino(2S)-pyrrolidinyl)methoxybe-
nzoate (430 mg, 1.1 mmol) in CH.sub.2Cl.sub.2 (10.0 ml) was added
TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product,
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (368 mg,
1.1 mmol), HOBt (162 mg, 1.2 mmol), and triethylamine (417 ml, 3.0
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (288
mg, 1.1 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1:1, v/v) as eluent to give
4-[trans-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-dimeth-
ylamino-(2S)-pyrrolidinyl]methoxy benzoate (530 mg, 78%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.94-1.99 (m, 1H),
2.48 (s, 9H), 3.06-3.12 (m, 1H), 3.33-3.38 (m, 1H), 3.60 (s, 2H),
3.68 (s, 3H), 3.69-3.80 (m, 1H), 3.88 (s, 3H), 4.13-4.20 (m, 2H),
4.56 (m, 1H), 6.76-7.00 (m, 5H), 7.22-7.34 (m, 3H), 7.92-8.00 (m,
3H), 8.17-8.19 (m, 1H). For HCl salt: IR (KBr) 3324, 2950, 2454,
1710, 1604, 1511, 1284 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta.2.30-2.40 (m, 2H), 2.77-2.80 (m, 6H), 3.60-3.75 (m, 2H),
3.75-3.85 (m, 8H), 4.00-4.22 (m, 3H), 4.50-4.75 (m, 1H), 6.75-7.43
(m, 7H), 7.89-8.09 (m, 4H), 8.87 (s, 1H), 8.91 (s, 1H); MS (FAB)
m/z 595 (M+H).sup.+; Anal. calcd for
C.sub.31H.sub.36N.sub.4O.sub.6Cl.1.0HCl.1.0H.sub.2O: C, 57.23; H,
6.04; N, 8.61; Cl, 10.90. Found: C, 57.43; H, 6.08; N, 8.38; Cl,
10.73.
[0990] To a stirred solution of methyl
4-[trans-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-4-dimet-
hylamino-(2S)-pyrrolidinyl]methoxybenzoate (190 mg, 0.32 mmol) in
THF (3.0 ml) and MeOH (2.0 ml) was added 1N NaOH (0.64 ml, 0.64
mmol). The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrate in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 159 (150 mg, 83%) as a white
crystalline solid. MW 581.06 mp 159-161.degree. C.; IR (KBr) 3318,
2938, 1604, 1531, 1438, 1340 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 2.10-2.40 (m, 8H), 2.50-2.70 (m, 2H), 3.85-3.90 (m, 5H),
4.02-4.18 (m, 3H), 4.30-4.60 (m, 1H), 6.75-7.43 (m, 7H), 7.86-8.09
(m, 4H), 8.86 (s, 1H), 8.91 (s, 1H); MS (FAB) m/z 581 (M+H).sup.+;
Anal. calcd for C.sub.30H.sub.33N.sub.4O.sub.6Cl.1.2H.sub.2O: C,
59.79; H, 5.92; N, 9.30. Found: C, 59.69; H, 5.93; N, 9.09.
Example 152
4-[cis-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-dimethyla-
mino-(2S)-pyrrolidinyl]methoxybenzoic acid
[0991] ##STR1782##
[0992] To a stirred solution of methyl
4-cis-1-tert-butoxycarbonyl-4-dimethylamino-(2S)-pyrrolidinyl)methoxybenz-
oate (1.2 g, 3.2 mmol) in CH.sub.2Cl.sub.2 (10.0 ml) was added TFA
(5.0 ml) at 0.degree. C. The reaction mixture was stirred at room
temperature for 0.5 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (278 mg, 1.0 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (335 mg,
1.0 mmol), HOBt (135 mg, 1.0 mmol), and triethylamine (417 ml, 3.0
mmol) in THF (4.0 ml) and MeCN (4.0 ml) was added EDC.HCl (288 mg,
1.5 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:1, v/v) as eluent to give methyl
4-[cis-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-dimethyl-
amino-(2S)-pyrrolidinyl]methoxybenzoate (500 mg, 84%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.98-2.50 (m, 1H),
2.26 (s, 3H), 2.25-2.40 (m, 1H), 2.58-2.65 (m, 1H), 3.20-3.30 (m,
1H), 3.60 (s, 2H), 3.64 (s, 3H), 3.80-3.90 (m, 1H), 3.88 (s, 3H),
4.18-4.20 (m, 1H), 4.42-4.46 (m, 2H), 6.72-7.00 (m, 4H), 7.20-7.35
(m, 5H), 7.91-7.94 (m, 3H), 8.18-8.21 (m, 1H).
[0993] To a stirred solution of methyl
4-[cis-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-4-dimethy-
lamino-(2S)-pyrrolidinyl]methoxybenzoate (250 mg, 0.42 mmol) in THF
(5.0 ml) and MeOH (3.0 ml) was added 1N NaOH (1.0 ml, 1.0 mmol).
The mixture was stirred at 70.degree. C. for 18 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 160 (170 mg, 70%) as a white crystalline
solid. MW 581.06 mp 165-167.degree. C.; IR (KBr) 3328, 1604, 1531,
1164, 1033 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.80-1.90
(m, 1H), 2.20-2.50 (m, 7H), 3.60-3.70 (m, 2H), 3.77-3.81 (m, 5H),
4.00-4.30 (m, 4H), 6.72-7.44 (m, 7H), 7.86-8.10 (m, 4H), 8.88-8.92
(m, 2H); MS (FAB) m/z 581 (M.sup.++1); Anal. calcd for
C.sub.30H.sub.33N.sub.4O.sub.6Cl.1.1H.sub.2O: C, 59.87; H, 6.06; N,
9.31. Found: C, 59.65; H, 5.76; N, 9.09.
Example 153
4-[cis-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-(2-naphth-
alenesulfonamido)-(2S)-pyrrolidinyl]methoxybenzoic acid
[0994] ##STR1783##
[0995] To a stirred solution of methyl
4-cis-1-tert-butoxycarbonyl-4-amino-(2S)-pyrrolidinyl)methoxy
benzoate (200 mg, 0.57 mmol) and TEA (317 ml, 2.3 mmol) in
CHCl.sub.3 (10.0 ml) was added (2-naphthyl)sulfonyl chloride (155
mg, 0.68 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 18 hr. The reaction mixture was concentrated
in vacuo. The residue was purified by column chromatography on
silica gel with n-hexane-EtOAc (2:1, v/v) as eluent to give methyl
4-(cis-1-tert-butoxycarbonyl-4-(2-naphthylsulfonamido)-(2S)-pyrrolidinyl)-
methoxybenzoate (240 mg, 78%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.25-1.45 (br s, 9H), 1.70-1.80 (m, 1H),
2.20-2.40 (m, 1H), 3.20-3.50 (m, 2H), 3.90 (s, 3H), 3.85-4.15 (m,
3H), 4.55-4.65 (, 1H), 6.90-7.10 (m, 2H), 7.58-8.04 (m, 8H), 8.43
(s, 1H).
[0996] To a stirred solution of methyl
4-(cis-1-tert-butoxycarbonyl-4-(2-naphthylsulfonamido)-(2S)-pyrrolidinyl)-
methoxybenzoate (240 mg, 0.44 mmol) in CH.sub.2Cl.sub.2 (5.0 ml)
was added TFA (5.0 ml) at 0.degree. C. The reaction mixture was
stirred at room temperature for 0.5 hr. The mixture was
concentrated in vacuo. Sat. NaHCO.sub.3 was added to the residue,
and extracted with CH.sub.2Cl.sub.2. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The
crude product was used to the subsequent reaction without further
purification. To a stirred solution of the crude product,
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (147 mg,
0.44 mmol), HOBt (59 mg, 0.44 mmol), and triethylamine (275 ml, 1.9
mmol) in THF (6.0 ml) and MeCN (6.0 ml) was added EDC.HCl (127 mg,
0.66 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:3, v/v) as eluent to give methyl
4-[cis-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-(2-napht-
halenesulfonamido)-(2S)-pyrrolidinyl]methoxybenzoate (200 mg, 65%)
as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.75-1.80 (m,
1H), 2.25-2.40 (m, 1H), 3.43 (s, 2H), 3.40-3.50 (m, 1H), 3.60 (s,
3H), 3.65-3.75 (m, 1H), 3.90 (s, 3H), 3.85-3.92 (m, 1H), 3.95-4.00
(m, 1H), 4.30-4.40 (m, 1H), 4.65-4.75 (m, 1H), 6.26 (d, J=9.3 Hz,
1H), 6.50 (d, J=8.3 Hz, 1H), 6.23 (s, 1H), 6.86 (d, J=8.8 Hz, 2H),
6.75-7.01 (m, 1H), 7.23-7.36 (m, 3H), 7.61-7.96 (m, 9H), 8.20 (d,
J=8.1 Hz, 1H), 8.43 (s, 1H).
[0997] To a stirred solution of methyl
4-[cis-1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenyl
acetyl]-4-(2-naphthalenesulfonamide)-(2S)-pyrrolidinyl]methoxybenzoate
(200 mg, 0.26 mmol) in THF (6.0 ml) and MeOH (3.0 ml) was added 1N
NaOH (0.5 ml, 0.5 mmol). The mixture was stirred at 70.degree. C.
for 18 hr. The mixture was concentrated in vacuo, water was added
thereto, and neutralized with 1N HCl. The resulting solid was
collected, washed with water, and dried in vacuo to give 161 (210
mg, quant) as a white crystalline solid. MW 743.22 mp
135-142.degree. C.; IR (KBr) 3332, 1685, 1604, 1531, 1421, 1159
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.75-1.85 (m, 1H),
2.05-2.15 (m, 1H), 3.05-3.15 (m, 1H), 3.47 (s, 2H), 3.60-3.80 (m,
2H), 3.73 (s, 3H), 4.05-4.20 (m, 3H), 6.51 (d, J=8.5 Hz, 1H),
6.74-7.04 (m, 5H), 7.27-7.31 (m, 1H), 7.43-7.45 (m, 2H), 7.66-8.17
(m, 9H), 8.46 (s, 1H), 8.91 (d, J=9.5 Hz, 1H); MS (FAB) m/z 743
(M.sup.++1); Anal. calcd for
C.sub.38H.sub.35N.sub.4O.sub.8ClS.0.5H.sub.2O: C, 60.67; H, 4.82;
N, 7.45; Cl, 4.26. Found: C, 60.77; H, 4.84; N, 7.21; Cl, 4.90.
Example 154
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-(2-mesitylenes-
ulfonamido-(2S)-pyrrolidinyl]methoxybenzoic acid
[0998] ##STR1784##
[0999] To a stirred solution of methyl
4-cis-1-tert-butoxycarbonyl-4-amino-(2S)-pyrrolidinyl)methoxy
benzoate (180 mg, 0.51 mmol) and TEA (283 ml, 2.0 mmol) in
CHCl.sub.3 (10.0 ml) was added (2-mesitylene)sulfonyl chloride (122
mg, 0.56 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 18 hr. The reaction mixture was concentrated
in vacuo. The residue was purified by column chromatography on
silica gel with n-hexane-EtOAc (3:1, v/v) as eluent to give methyl
4-(cis-1-tert-butoxycarbonyl-4-(2-mesitylenesulfonamido-(2S)-pyrrolidinyl-
)methoxy benzoate (170 mg, 62%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.40 (s, 9H), 1.85-1.95 (m, 1H), 2.29 (s, 3H),
2.35-2.45 (m, 1H), 2.62 (s, 6H), 3.80-4.15 (m, 3H), 3.89 (s, 3H),
3.50-3.65 (m, 1H), 6.94 (s, 2H), 6.94-7.00 (m, 2H), 7.99 (d, J=8.8
Hz, 2H).
[1000] To a stirred solution of methyl
(cis-1-tert-butoxycarbonyl-4-(2-mesitylenesulfonamido)-(2S)-pyrrolidinyl)-
methoxybenzoate (170 mg, 0.32 mmol) in CH.sub.2Cl.sub.2 (5.0 ml)
was added TFA (5.0 ml) at 0.degree. C. The reaction mixture was
stirred at room temperature for 0.5 hr. The mixture was
concentrated in vacuo. Sat. NaHCO.sub.3 was added to the residue,
and extracted with CH.sub.2Cl.sub.2. The extract was washed with
brine dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The
crude product was used to the subsequent reaction without further
purification. To a stirred solution of the crude product,
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (121 mg,
0.32 mmol), HOBt (43 mg, 0.32 mmol), and triethylamine (139 ml, 1.0
mmol) in THF (5.0 ml) and MeCN (5.0 ml) was added EDC.HCl (91 mg,
0.48 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:4, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-(2-mesitylene-
sulfonamido)-(2S)-pyrrolidinyl]methoxybenzoate (210 mg, 83%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.85-1.90 (m, 1H),
1.95-2.05 (m, 1H), 2.32 (s, 3H), 2.60 (s, 6H), 3.40-3.50 (m, 3H),
3.60-3.70 (ma, 2H), 3.68 (s, 3H), 3.89 (s, 3H), 3.96-3.99 (m, 1H),
3.35-3.45 (m, 1H), 3.70-3.75 (m, 1H), 6.00 (d, J=9.5 Hz, 1H),
6.57-7.08 (m, 9H), 7.29-7.34 (m, 1H), 7.51-7.53 (m, 1H), 7.92-7.96
(m, 3H), 8.15 (d, J=6.8 Hz, 1H).
[1001] To a stirred solution of methyl
4-[1-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetyl]-4-(2-mesitylene-
sulfonamido-(2S)-pyrrolidinyl]methoxybenzoate (210 mg, 0.26 mmol)
in THF (5.0 ml) and MeOH (3.0 ml) was added 1N NaOH (0.47 ml, 0.47
mmol). The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 162 (180 mg, 87%) as a white
crystalline solid. MW 779.70 mp 130-132.degree. C.; IR (KBr) 3332,
1689, 1604, 1529, 1155 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.70-1.85 (m, 1H), 2.02-2.12 (m, 1H), 2.25 (s, 3H), 2.52
(s, 6H), 3.05-3.12 (m, 1H), 3.48 (s, 2H), 3.60-3.70 (m, 2H), 3.77
(s, 3H), 3.90-4.20 (m, 3H), 6.59 (d, J=8.3 Hz, 1H), 6.77-6.80 (m,
1H), 6.95-7.01 (m, 4H), 7.31-7.35 (m, 1H), 7.60 (d, J=8.1 Hz, 1H),
7.86-7.97 (m, 5H), 8.72-8.76 (m, 1H), 8.89-8.93 (m, 1H); MS (FAB)
m/z 779 (M.sup.+), 781 (M.sup.++2); Anal. calcd for
C.sub.37H.sub.39N.sub.4O.sub.8SBr.0.5H.sub.2O: C, 56.35; H, 5.11;
N, 7.10; Br, 10.13. Found: C, 56.39; H, 5.07; N, 6.89; Br,
10.25.
Example 155
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-dansylamino-(2-
S)-pyrrolidinyl]methoxybenzoic acid
[1002] ##STR1785##
[1003] To a stiffed solution of methyl
4-(cis-1-tert-butoxycarbonyl-4-amino-(2S)-pyrrolidinyl)methoxy
benzoate (180 mg, 0.51 mmol) and TEA (283 ml, 2.0 mmol) in
CHCl.sub.3 (10.0 ml) was added dansyl chloride (155 mg, 0.68 mmol)
at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. The reaction mixture was concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (2:1, v/v) as eluent to give methyl
4-(cis-1-tert-butoxycarbonyl-4-dansylamino-(2S)-pyrrolidinyl)methoxy]benz-
oate (200 mg, 73%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.34 (s, 9H), 1.50-1.60 (m, 1H), 2.15-2.25 (m, 1H), 2.87
(s, 6H), 3.15-3.22 (m, 1H), 3.35-3.45 (m, 1H), 3.48-3.52 (m, 1H),
3.80-4.10 (m, 3H), 3.91 (s, 3H), 7.01]-7.25 (m, 4H), 7.50-7.53 (m,
1H), 8.03 (d, J=8.7 Hz, 2H), 8.16 (d, J=8.5 Hz, 1H), 8.28 (d, J=7.1
Hz, 1H), 8.53 (d, J=8.5 Hz, 1H).
[1004] To a stirred solution of methyl
4-(cis-1-tert-butoxycarbonyl-4-dansylamino-(2S)-pyrrolidinyl)methoxybenoa-
te (200 mg, 0.34 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was added TFA
(5.0 ml) at 0.degree. C. The reaction mixture was stirred at room
temperature for 0.5 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stiffed solution of the crude product,
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (129 mg,
0.34 mmol), HOBt (46 mg, 0.34 mmol), and triethylamine (142 ml, 1.0
mmol) in THF (5.0 ml) and MeCN (5.0 ml) was added EDC.HCl (98 mg,
0.51 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:4, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4]dansylamino-(-
2S)-pyrrolidinyl]methoxybenzoate (250 mg, 88%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.55-1.65 (m, 1H), 2.15-2.25 (m,
1H), 2.87 (s, 6H), 3.20-3.35 (m, 3H), 3.50-3.55 (m, 1H), 3.67 (s,
3H), 3.78-3.81 (m, 1H), 3.88-3.93 (m, 1H), 3.91 (s, 3H), 4.28-4.31
(m, 1H), 4.65-4.70 (m, 1H), 6.35 (d, J=9.5 Hz, 1H), 6.54 (d, J=8.5
Hz, 1H), 6.63 (s, 1H), 6.90-7.13 (m, 6H), 7.22-7.31 (m, 2H),
7.50-7.56 (m, 2H), 7.88 (d, J=8.0 Hz, 1H), 7.99 (d, J=9.0 Hz, 1H),
8.13-8.16 (m, 2H), 8.27 (d, J=7.6 Hz, 1H), 8.56 (d, J=8.3 Hz,
1H).
[1005] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-dansylamino-2-
S-pyrrolidinyl]methoxybenzoate (250 mg, 0.29 mmol) in THF (5.0 ml)
and MeOH (3.0 ml) was added 1N NaOH (0.52 ml, 0.52 mmol). The
mixture was stirred at 70.degree. C. for 24 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 163 (230 mg, 94%) as a green crystalline
solid. MW 830.74 mp 138-141.degree. C.; IR (KBr) 3340, 2940, 1604,
1527, 1421, 1162, 1145 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.70-1.80 (m, 1H), 1.98-2.06 (m, 1H), 2.81 (s, 6H),
3.00-3.10 (m, 1H), 3.39-3.40 (m, 2H), 3.50-3.80 (m, 3H), 3.76 (s,
3H), 3.90-4.15 (m, 2H), 6.52 (d, J=9.0 Hz, 1H), 6.73-7.00 (m, 4H),
7.22-7.35 (m, 2H), 7.55-7.64 (m, 3H), 7.83-8.48 (m, 8H), 8.71-8.76
(m, 1H), 8.88-8.92 (m, 1H); MS (FAB) m/z 830 (M.sup.+), 832
(M.sup.++2); Anal. calcd for
C.sub.40H.sub.40N.sub.5O.sub.5O.sub.8BrS.0.7H.sub.2O: C, 56.97; H,
95; N, 8.30; Br, 9.47. Found: C, 57.06; H, 4.86; N, 7.98; Br,
9.66.
Example 156
4-[4-methanesulfonamido-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylace-
tyl]-(2S)-pyrrolidinyl]methoxybenzoic acid
[1006] ##STR1786##
[1007] To a stirred solution of methyl
4-cis-4-amino-1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxy
benzoate (180 mg, 0.51 mmol) and TEA (283 ml, 2.0 mmol) in
CHCl.sub.3 (10.0 ml) was added methanesulfonyl chloride (88 mg,
0.77 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. The reaction mixture was concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1:1, v/v) as eluent to give methyl
4-(cis-1-tert-butoxycarbonyl-4-methanesulfonamido-(2S)-pyrrolidinyl)metho-
xybenzoate (150 mg, 69%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.45 (s, 9H), 1.98-2.08 (m, 1H), 2.52-2.65 (m,
1H), 2.99 (s, 3H), 3.40-3.50 (m, 1H), 3.55-3.80 (m, 1H), 3.89 (s,
1H), 4.00-4.70 (m, 4H), 6.98-7.00 (m, 2H), 8.00 (d, J=8.8 Hz,
2H).
[1008] To a stirred solution of methyl
4-cis-1-tert-butoxycarbonyl-4-methanesulfonamido-(2S)-pyrrolidinyl)methox-
ybenzoate (150 mg, 0.43 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was
added TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product,
4-[N'-2-bromophenyl)uredio]-3-methoxyphenylacetic acid (163 mg,
0.42 mmol), HOBT (58 mg, 0.43 mmol), and triethylamine (179 ml, 1.3
mmol) in THF (5.0 ml) and MeCN (5.0 ml) was added EDC.HCl (144 mg,
0.75 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:4, v/v) to EtOH-EtOAc (10%, v/v) as eluent to
give methyl
4-[4-methanesulfonamido-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(2S)-pyrrolidinyl]methoxybenzoate (210 mg, 73%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.95-2.00 (m, 1H),
2.55-2.65 (m, 1H), 2.17 (s, 3H), 3.55-3.70 (m, 1H), 3.60 (s, 2H),
3.67 (s, 3H), 3.85-3.90 (m, 1H), 3.89 (s, 3H), 3.95-4.18 (m, 2H),
4.45-4.55 (m, 1H), 4.70-4.80 (m, 1H), 5.87 (d, J=9.3 Hz, 1H),
6.73-6.95 (m, 5H), 7.09 (s, 2H), 7.28-7.33 (m, 1H), 7.51 (d, J=8.0
Hz, 1H), 7.93-7.97 (m, 3H), 8.13 (d, J=8.3 Hz, 1H).
[1009] To a stirred solution of methyl
4-[4-methanesulfonamido-1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylac-
etyl]-(2S)-pyrrolidinyl]methoxybenzoate (210 mg, 0.3 mmol) in THF
(5.0 ml) and MeOH (3.0 ml) was added 1N NaOH (0.8 ml, 0.8 mmol).
The mixture was stirred at 70.degree. C. for 24 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 164 (170 mg, 83%) as a white crystalline
solid. MW 675.55 mp 125-128.degree. C.; IR (KBr) 3353, 1689, 1604,
1529, 1419, 1155 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.88-2.00 (m, 1H), 2.35-2.45 (m, 1H), 2.96 (m, 3H), 3.15-3.23 (m,
1H), 3.60 (s, 2H), 3.50-3.70 (m, 1H), 3.78 (s, 3H), 3.80-3.90 (m,
1H), 3.95-4.05 (m, 1H), 4.10-4.30 (m, 2H), 6.71-7.03 (m, 4H), 7.32
(m, 1H), 7.45 (d, J=6.8 Hz, 1H), 7.60 (d, J=7.8 Hz, 1H), 7.87-7.95
(m, 4H), 8.74 (s, 1H), 8.92 (s, 1H); MS (FAB) m/z 675 (M.sup.+),
677 (M.sup.++2); Anal. calcd for
C.sub.29H.sub.31N.sub.4O.sub.8BrS.0.6H.sub.2O: C, 50.75; H, 4.73;
N, 8.16. Found: C, 51.04; H, 4.62; N, 7.79.
Example 157
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-octahydroi-
ndolylmethoxy]benzoic acid
[1010] ##STR1787##
[1011] To a stirred solution of octahydroindole-(2S)-carboxylic
acid (3.00 g, 17.7 mmol) in dioxane (20 ml) was added 1 N NaOH (45
ml) and the solution was stirred at 0.degree. C. To the mixture was
added (Boc).sub.2O (4.26 g, 19.5 mmol) in dioxane (25 ml) at
0.degree. C. and the reaction mixture was stirred at room
temperature for 1 day. The mixture was acidified with 1 N HCl and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated to give 1-(tert-butoxy
carbonyl)octahydroindole-(2S)-carboxylic acid (4.78 g, q.y.) as a
colorless solid. mp 130-132.degree. C.; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.10-1.46 (series of s and m, total 14H), 1.65-1.76 (m,
3H), 1.90-2.18 (m, 2H), 2.26-2.35 (m, 1H), 3.77-3.86 (m, 1H),
4.22-4.34 (m, 1H); MS (ESI) m/z 270 (M.sup.++1).
[1012] To a cooled (0.degree. C.), stirred solution of
1-(tert-butoxycarbonyl)octahydroindole-(2S)-carboxylic acid (1.00
g, 3.71 mmol) in THF (10 ml) was added BH.sub.3DMS (530 ml, 5.59
mmol) and the reaction mixture was stirred at room temperature
overnight. The mixture was quenched by H.sub.2O and extracted with
EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as
eluent to give 1-(tert-butoxycarbonyl)octahydroindole-(2S)-methanol
(940 mg, 99%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.05-1.30 (m, 4H), 1.47 (s, 9H), 1.49-1.74 (m, 4H), 1.82-1.93 (m,
3H), 2.19-2.26 (m, 1H), 3.56-3.61 (m, 1H), 3.70-3.75 (m, 2H),
3.94-3.96 (m, 1H); MS (FAB) m/z 256 (M.sup.++1).
[1013] To a cooled (0.degree. C.), stirred solution of methyl
4-hydroxybenzoate (560 mg, 3.68 mmol),
1-(tert-butoxycarbonyl)octahydroindole-(2S)-methanol (940 mg, 3.68
mmol) and Ph.sub.3P (1.16 g, 4.42 mmol) in THF (20 ml) was added
DIAD (870 ml, 4.42 mmol) and the reaction mixture was heated under
reflux for 8 hr. After cooled to room temperature, the mixture was
evaporated. The residue was purified by column chromatography on
silica-gel with n-hexane-EtOAc (5:1, v/v) as eluent to give methyl
4-[1-(tert-butoxycarbonyl)-(2S)-octahydroindolylmethoxy]benzoate
(1.16 g, 81%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.14-1.47 (series of s and m, total 13H), 1.60-2.13 (series of m,
total 6H), 2.22-2.28 (m, 1H), 3.75-3.91 (series of s and m, total
4H), 4.06-4.18 (m, 2H), 4.37 (m, 1H), 6.94-6.96 (m, 2H), 7.96-7.98
(m, 2H); MS (FAB) m/z 390 (M.sup.++1).
[1014] To a stirred solution of methyl
4-[1-(tert-butoxycarbonyl)-(2S)-octahydroindolylmethoxy]benzoate
(1.16 g, 2.98 mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (10
ml) and the reaction mixture was stirred at room temperature for 2
hr. The mixture was concentrated in vacuo, made basic by sat.
NaHCO.sub.3, and extracted with CHCl.sub.3. The extract was washed
with brine, dried over K.sub.2CO.sub.3, and evaporated to give
methyl 4-[(2)-octahydroindolylmethoxy]benzoate (860 mg, q.y.) as a
brown oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.23-1.78 (series of m,
total 10H), 2.00-2.09 (m, 2H), 3.14-3.18 (m, 1H), 3.55-3.62 (m,
1H), 3.88 (s, 3H), 3.96-4.06 (m, 2H), 6.92 (d, J=9.1 Hz, 2H), 7.97
(d, J=9.1 Hz, 2H); MS (FAB) m/z 290 (M.sup.++1).
[1015] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (298 mg,
0.95 mmol), methyl 4-[(2S)-octahydroindolylmethoxy]benzoate (274
mg, 0.95 mmol), EDC.HCl (218 mg, 1.14 mmol), HOBt (154 mg, 1.14
mmol), Et.sub.3N (160 ml, 1.15 mmol) in THF (7 ml) was stirred at
room temperature overnight. The mixture was diluted with H.sub.2O
and extracted with EtOAc. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1 to
50:1, v/v) as eluent to give methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-octahydro-
indolyl methoxy]benzoate (532 mg, 96%) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.13-2.05 (series of m, total 9H), 2.14-2.24
(m, 2H), 2.26 (s, 3H), 3.60 (s, 2H), 3.62 (s, 3H), 3.80-3.85 (m,
1H), 3.88 (s, 3H), 4.27-4.37 (m, 3H), 6.62 (s, 1H), 6.74-6.76 (m,
2H), 6.91 (d, J=8.8 Hz, 2H), 7.09-7.13 (m, 1H), 7.20-7.24 (m, 3H),
7.55 (d, J=7.8 Hz, 1H), 7.94 (d, J=8.8 Hz, 2H), 8.04 (d, J=7.8 Hz,
1H); MS (FAB) m/z 586 (M.sup.++1).
[1016] To a stirred solution of methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-(2S)-octahydroindolylmethoxy]benzoate (532 mg, 0.91 mmol)
in THF (5 ml) was added 0.5 N NaOH (5 ml) and the reaction mixture
was heated under reflux for 3 hr. After cooled to room temperature,
the mixture was poured into ice-1 N HCl, and the resulting
precipitate was collected. The crude solid was recrystallized from
MeOH--CHCl.sub.3-[PE to give 165 (278 mg, 54%) as a white
crystalline powder. MW 571.66 mp 130-134.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.16-2.10 (series of m, total 9H), 2.15-2.30
(series of s and m, total 4H), 3.55-3.79 (m, 3H), 3.81 (s, 3H),
3.90-3.95 (m, 1H), 4.17-4.23 (m, 2H), 4.34-4.36 (m, 1H), 6.72-6.74
(m, 1H), 6.87-6.88 (m, 1H), 6.91-6.95 (m, 1H), 7.03 (d, J=8.8 Hz,
2H), 7.10-7.16 (m, 2H), 7.78-7.80 (m, 1H), 7.87 (d, J=8.8 Hz, 2H),
7.98-8.00 (m, 1H), 8.45 (s, 1H), 8.54 (s, 1H), 12.61 (br s, 1H); MS
(FAB) m/z 572 (M.sup.++1); Anal. Calcd for
C.sub.33H.sub.37N.sub.3O.sub.6.1/4H.sub.2O: C, 68.79; H, 6.56; N,
7.29. Found: C, 68.70; H, 6.82; N, 6.97.
Example 158
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(2S)-octahydroi-
ndolylmethoxy]benzoic acid
[1017] ##STR1788##
[1018] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (307 mg,
0.92 mmol), methyl 4-[(2S)-octahydroindolylmethoxy]benzoate (265
mg, 0.92 mmol), EDC.HCl (211 mg, 1.10 mmol), HOBt (148 mg, 1.10
mmol), and Et.sub.3N (153 ml, 1.10 mmol) in THF (7 ml) was stirred
at room temperature overnight. The mixture was diluted with
H.sub.2O and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(2S)-octahydro-
indolyl methoxy]benzoate (550 mg, 99%) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.15-2.02 (series of m, total 9H), 2.17-2.33
(m, 2H), 3.58 (s, 3H), 3.62 (s, 2H), 3.84-3.90 (series of s and m,
total 4H), 4.06-4.40 (m, 3H), 6.71-6.74 (m, 2H), 6.88-7.00 (m, 3H),
7.21-7.30 (m, 2H), 7.62 (s, 2H), 7.91-7.95 (m, 3H), 8.17-8.20 (m,
1H); MS (FAB) m/z 606 (M.sup.++1).
[1019] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(2S)-octahydro-
indolylmethoxy]benzoate (550 mg, 0.91 mmol) in THF (5 ml) was added
0.5 N NaOH (5 ml) and the reaction mixture was heated under reflux
for 5 hr. After cooled to room temperature, the mixture was poured
into ice-1 N HCl, and the resulting precipitate was collected. The
crude solid was recrystallized from MeOH--CHCl.sub.3--IPE to give
166 (286 mg, 53%) as a white crystalline powder. MW 388.29 mp
133-136.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.16-2.10
(series of m, total 10H), 2.24-2.27 (m, 1H), 3.55-3.75 (m, 2H),
3.80 (s, 3H), 3.90-3.96 (m, 1H), 4.17-4.23 (m, 2H), 4.34-4.36 (m,
1H), 6.73-6.75 (m, 1H), 6.88 (d, J=1.5 Hz, 1H), 6.99-7.05 (m, 3H),
7.25-7.30 (m, 1H), 7.43 (dd, J=1.5, 8.1 Hz, 1H), 7.87-7.89 (m, 2H),
7.95 (d, J=8.1 Hz, 1H), 8.08 (dd, J=1.5, 8.3 Hz, 1H), 8.88 (s, 1H),
8.92 (s, 1H), 12.61 (br s, 1H); MS (FAB) m/z 592 (M.sup.++1); Anal.
Calcd for C.sub.32H.sub.34ClN.sub.3O.sub.6.1/4H.sub.2O: C, 64.42;
H, 5.83; N, 7.04; Cl, 5.94. Found: C, 64.55; H, 6.09; N, 6.64; Cl,
5.93.
Example 159
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(2S)-octahydroin-
dolylmethoxy]benzoic acid
[1020] ##STR1789##
[1021] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (457 mg,
1.21 mmol), methyl 4-[(2S)-octahydroindolylmethoxy]benzoate (320
mg, 1.21 mmol), EDC.HCl (277 mg, 1.44 mmol), HOBt (196 mg, 1.45
mmol), and Et.sub.3N (200 ml, 1.43 mmol) in THF (7 ml) was stirred
at room temperature overnight. The mixture was diluted with
H.sub.2O and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(2S)-octahydroi-
ndolyl methoxy]benzoate (423 mg, 54%) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.15-1.89 (series of m, total 8H), 1.96-2.02
(m, 1H), 2.16-2.32 (m, 2H), 3.63 (s, 2H), 3.65 (s, 3H), 3.82-3.86
(m, 1H), 3.88 (s, 3H), 4.30-4.39 (m, 3H), 6.75-6.77 (m, 2H),
6.88-0.93 (m, 3H), 7.24-7.31 (m, 1H), 7.37-7.50 (m, 3H), 7.91-7.99
(m, 3H), 8.12-8.15 (m, 1H); MS (FAB) m/z 650 (M.sup.++1).
[1022] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(2S)-octahydroindolylmethoxy]benzoate (420 mg, 0.65 mmol)
in THF (5 ml) was added 0.5 N NaOH (5 ml) and the reaction mixture
was heated under reflux for 5 hr. After cooled to room temperature,
the mixture was poured into ice-1 N HCl, and the resulting
precipitate was collected. The crude solid was recrystallized from
MeOH--CHCl.sub.3--IPE to give 167 (197 mg, 48%) as a white
crystalline powder. MW 636.53 mp 118-123.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.16-2.27 (series of m, total 10H),
3.56-3.75 (m, 3H), 3.81 (s, 3H), 3.90-3.96 (m, 1H), 4.17-4.23 (m,
2H), 4.34-4.36 (m, 1H), 6.73-6.75 (m, 1H), 6.88-7.05 (m, 4H),
7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.96 (m, 4H), 8.73 (s,
1H), 8.91 (s, 1H), 12.64 (br s, 1H); MS (FAB) m/z 636 (M.sup.++1);
Anal. Calcd for C.sub.32H.sub.34BrN.sub.3O.sub.6.1/4H.sub.2O: C,
59.96; H, 5.42; N, 6.55; Br, 12.46. Found: C, 60.12; H, 5.86; N,
6.09; Br, 12.47.
Example 160
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetyl-4-thiazolidinyl-
]methoxybenzoic acid
[1023] ##STR1790##
[1024] To a stirred solution of thiazolidine-4-caboxylic acid (5.0
g, 37.6 mmol) in DMF (50.0 ml) was added (Boc).sub.2O (9.8 g, 45.1
mmol) and TEA (8.0 ml). The reaction mixture was stirred at room
temperature for 18 hr. Water was added to the mixture and extracted
with EtOAc. The organic layer was washed with water, then dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with EtOAc-n-hexane
(1:3 v/v) as eluent to give
3-tert-butoxycarbonylthiazolidine-4-carboxylic acid (6.5 g, 74%) as
a white crystalline solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.49
(br s, 9H), 3.20-3.30 (m, 2H), 4.09-4.87 (m, 3H).
[1025] To a stirred solution of
3-tert-butoxycarbonylthiazolidine-4-carboxylic acid (2.3 g, 10.0
mmol) in THF (30 ml) was added BH.sub.3-THF (1.0 M solution in THF,
20.0 ml, 20.0 mmol) at 0.degree. C. After stirred at room
temperature for 1.0 h, the reaction mixture was heated under reflux
for 1.0 hr. After cooled, the mixture was concentrated in vacuo.
Water was added thereto at 0.degree. C., and extracted with EtOAc.
The extract was washed with water, then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo to give
3-tert-butoxycarbonyl-5-hydroxymethylthiazolidine (2.0 g, quant) as
a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.48 (s, 9H),
2.80-2.85 (m, 1H), 3.13-3.17 (m, 1H), 3.20-3.30 (m, 1H), 3.64-3.70
(m, 2H), 4.34 (br s, 1H), 4.60 (br s, 1H).
[1026] To a stirred solution of
3-tert-butoxycarbonyl-5-hydroxymethylthiazolidine (1.9 g, 8.7
mmol), methyl 4-hydroxybenzoate (1.3 g, 8.7 mmol), and Ph.sub.3P
(3.2 g, 12.2 mmol) in THF (10 ml) was added DIAD (2.2 g, 10.4 mmol)
at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. The mixture was concentrated in vacuo. The
residue was purified by column chromatography on silica gel with
EtOAc-n-hexane (1:9, v/v) as eluent to give methyl
43-tert-butoxycarbonyl-4-thiazolidinyl)methoxybenzoate (1.6 g, 52%)
as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.49 (s,
9H), 3.11-3.19 (m, 2H), 3.88 (s, 3H), 4.04-4.31 (m, 3H), 4.61 (m,
2H), 6.96 (d, J=8.8 Hz, 2H), 7.98 (d, J=8.8 Hz, 2H).
[1027] To a stirred solution of methyl
4-(3-tert-butoxycarbonyl-4-thiazolidinyl)methoxybenzoate (440 mg,
1.25 mmol) in CH.sub.2Cl.sub.2 (6 ml) was added TFA (3 ml) at
0.degree. C. The reaction mixture was stirred at room temperature
for 0.5 hr. The mixture was concentrated in vacuo. Sat. NaHCO.sub.3
was added to the residue, and extracted with CH.sub.2Cl.sub.2. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The crude product was used to the subsequent
reaction without further purification. To a stirred solution of the
crude product (0.6 mmol), 3-methoxy-4-[N'-(2-methyl
phenyl)uredio]phenylacetic acid (188 mg, 0.6 mmol), HOBt (81 mg,
0.6 mmol), and triethylamine (280 ml, 2.0 mmol) in THF (5 ml) and
MeCN (5 ml) was added EDC.HCl (173 mg, 0.9 mmol) at 0.degree. C.
The reaction mixture was stirred at room temperature for 16 hr, and
concentrated in vacuo. Water was added to the residue, and
extracted with EtOAc. The extract was washed with sat. NaHCO.sub.3,
2-M citric acid, and sat. NaHCO.sub.3, then dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with n-hexane-EtOAc
(1:3, v/v) as eluent to give methyl
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetyl-4-thiazolidiny-
l]methoxybenzoate (340 mg, quant) as an amorphous solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 2.31 (s, 3H), 3.15-3.16 (m, 2H),
3.67-3.69 (m, 5H), 3.88 (s, 3H), 4.09-4.14 (m, 2H), 4.22-4.90 (m,
3H), 6.30 (m, 1H), 6.74-6.96 (m, 4H), 7.11-7.25 (m, 4H), 7.49-7.51
(m, 1H), 7.95-8.12 (m, 3H).
[1028] To a stirred solution of methyl
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetyl-4-thiazolidiny-
l]methoxybenzoate (340 mg, 0.62 mmol) in THF (5.0 ml) and EtOH (3.0
ml) was added 1N NaOH (0.62 ml, 0.62 mmol). The mixture was stirred
at 70.degree. C. for 18 hr. The mixture was concentrated in vacuo,
water was added thereto, and neutralized with 1N HCl. The resulting
solid was collected, washed with water, and dried in vacuo to give
168 (290 mg, 88%) as a white crystalline solid. MW 535.62 mp
125-128.degree. C.; IR (KBr) 3357, 2937, 1604, 1533, 1419, 1253,
1166, 1033, 773 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25
(s, 3H), 3.05-3.20 (m, 2H), 3.71, 3.83, and 3.85 (each s, total
5H), 4.03-4.15 (m, 3H), 4.52-4.76 (m, 2H), 6.15-6.17 (m, 7H),
7.78-8.30 (m, 4H), 8.30 (m, 1H), 8.56 (m, 1H); MS (FAB) m/z 536
(M.sup.++1); Anal. calcd for
C.sub.28H.sub.29N.sub.3O.sub.6S.0.5H.sub.2O: C, 61.75; H, 5.55; N,
7.72. Found: C, 61.72; H, 5.55; N, 7.49.
Example 161
4-[3-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-thiazolidinyl-
]methoxybenzoic acid
[1029] ##STR1791##
[1030] To a stirred solution of methyl
4-(3-tert-butoxycarbonyl-4-thiazolidinyl)methoxybenzoate (600 mg,
1.7 mmol) in CH.sub.2Cl.sub.2 (6.0 ml) was added TFA (6.0 ml) at
0.degree. C. The reaction mixture was stirred at room temperature
for 0.5 hr. The mixture was concentrated in vacuo. Sat. NaHCO.sub.3
was added to the residue, and extracted with CH.sub.2Cl.sub.2. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The crude product was used to the subsequent
reaction without further purification. To a stirred solution of the
crude product, 4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic
acid (570 mg, 1.7 mmol), HOBt (230 mg, 1.7 mmol), and triethylamine
(709 ml, 5.1 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added
EDC.HCl (490 mg, 2.55 mmol) at 0.degree. C. The reaction mixture
was stirred at room temperature for 16 hr, and concentrated in
vacuo. Water was added to the residue, and extracted with EtOAc.
The extract was washed with sat NaHCO.sub.3, 2-M citric acid, and
sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (1:1, v/v) as
eluent to give methyl
4-[3-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-thiazolidiny-
l]methoxybenzoate (900 mg, 93%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 3.15-3.18 (m, 2H), 3.70 (s, 2H), 3.78 (s, 3H),
3.86 (s, 3H), 4.09-4.93 (m, 5H), 6.80-7.01 (m, 5H), 7.19-7.35 (m,
4H), 7.94-8.18 (m, 4H).
[1031] To a stirred solution of methyl
4-[3-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-4-thiazolidiny-
l]methoxybenzoate (900 mg, 1.6 mmol) in THF (8.0 ml) and MeOH (4.0
ml) was added 1N NaOH (3.1 ml, 3.1 mmol). The mixture was stirred
at 70.degree. C. for 24 hr. The mixture was concentrated in vacuo,
water was added thereto, and neutralized with 1N HCl. The resulting
solid was collected, washed with water, and dried in vacuo to give
169 (780 mg, 89%) as a white crystalline solid. MW 556.03 mp
126-129.degree. C.; IR (K]Br) 3343, 2937, 1604, 1531, 1421, 1245,
1166, 1035, 752 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
3.06-3.24 (m, 2H), 3.72-3.85 (m, 5H), 4.02-4.27 (m, 3H), 4.53-4.76
(m, 2H), 6.74-7.44 (m, 7H), 7.87-8.30 (m, 4H), 8.89-8.95 (m, 2H);
MS (FAB) m/z 556 (M.sup.++1); Anal. calcd for
C.sub.27H.sub.27N.sub.3O.sub.6ClS.0.7H.sub.2O: C, 56.93; H, 5.03;
N, 7.38; Cl, 6.22. Found: C, 56.89; H, 4.84; N, 7.42; Cl, 6.35.
Example 162
4-[3-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-thiazolidinyl]-
methoxybenzoic acid
[1032] ##STR1792##
[1033] To a stirred solution of methyl
4-(3-tert-butoxycarbonyl-4-thiazolidinyl)methoxybenzoate (560 mg,
1.6 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was added TFA (5.0 ml) at
0.degree. C. The reaction mixture was stirred at room temperature
for 0.5 hr. The mixture was concentrated in vacuo. Sat. NaHCO.sub.3
was added to the residue, and extracted with CH.sub.2Cl.sub.2. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The crude product was used to the subsequent
reaction without further purification. To a stirred solution of the
crude product, 4-[N'-(2-bromophenyl)uredio]-3-methoxy phenylacetic
acid (599 mg, 1.6 mmol), HOBt (213 mg, 1.6 mmol), and triethylamine
(659 ml, 4.7 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added
EDC.HCl (455 mg, 2.4 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 16 hr, and concentrated in vacuo.
Water was added to the residue, and extracted with EtOAc. The
extract was washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (2:3, v/v) as eluent to give methyl
4-[3-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-thiazolidinyl-
]methoxybenzoate (870 mg, 89%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 3.00-3.20 (m, 3H), 3.70 (s, 2H), 3.81 (s, 3H),
3.88 (s, 3H), 4.09-4.23 (m, 1H), 4.42 (d, J=8.5 Hz, 1H), 4.59 (d,
J=8.5 Hz, 1H), 4.70-4.92 (m, 1H), 6.81-7.53 (m, 9H), 7.95-8.15 (m,
4H).
[1034] To a stirred solution of methyl
4-[3-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-4-thiazolidinyl-
]methoxybenzoate (870 mg, 1.4 mmol) in THF (8.0 ml) and MeOH (8.0
ml) was added 1N NaOH (2.8 ml, 2.8 mmol). The mixture was stirred
at 70.degree. C. for 18 hr. The mixture was concentrated in vacuo,
water was added thereto, and neutralized with 1N HCl. The resulting
solid was collected, washed with water, and dried in vacuo to give
170 (740 mg, 87%) as a white crystalline solid. MW 600.48 mp
125-133.degree. C.; IR (KBr) 3332, 2935, 1604, 1527, 1421, 1245,
1166, 1027, 750 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
3.01-3.25 (m, 2H), 3.72-3.85 (m, 5H), 4.02-4.30 (m, 2H), 4.54 (d,
J=8.8 Hz, 1H), 4.74-4.87 (m, 2H), 6.76-7.07 (m, 5H), 7.30-7.34 (m,
1H), 7.59 (d, J=8.1 Hz, 1H), 7.86-7.98 (m, 4H), 8.74 (s, 1H),
8.92-8.94 (s, 1H); MS (FAB) m/z 600 (M.sup.++1); Anal. calcd for
C.sub.27H.sub.26N.sub.3O.sub.6BrS.0.3H.sub.2O: C, 53.52; H, 4.43;
N, 6.94. Found: C, 53.54; H, 4.45; N, 6.80.
Example 163
cis-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolid-
inyl]methylamino]cyclohexanecarboxylic acid
[1035] ##STR1793##
[1036] To a stirred solution of 2S-pyrrolidinemethanol (2.0 g, 20.0
mmol), 3-methoxy-4-[N'-(2-methyl phenyl)uredio]phenylacetic acid
(6.28 g, 20.0 mmol), HOBt (71 mg, 0.53 mmol), and triethylamine
(5.5 ml, 40.0 mmol) in THF (50.0 ml) and MeCN (40.0 ml) was added
EDC.HCl (5.7 g, 30.0 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 16 hr, and concentrated in vacuo.
Water was added to the residue, and extracted with EtOAc. The
organic layer was washed with sat. NaHCO.sub.3, 2-M citric acid,
and sat. NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with EtOAc to MeOH--CH.sub.2Cl.sub.2
(1:9, v/v) as eluent to give
1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2S-pyrrolidinemet-
hanol (7.0 g, 89%) as a white crystalline solid. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.54-1.58 (m, 1H), 1.80-2.04 (m, 3H), 2.27 (s,
3H), 3.42-3.46 (m, 1H), 3.54-3.65 (m, 2H), 3.62 (s, 2H), 3.69 (s,
3H), 4.21-4.23 (m, 1H), 5.04 (m, 1H), 6.68-6.79 (m, 3H), 7.09-7.31
(m, 4H), 7.52 (d, J=7.8 Hz, 1H), 8.07 (d, J=8.0 Hz, 1H).
[1037] To a stirred solution of oxalyl chloride (0.3 ml, 3.3 mmol)
in CH.sub.2Cl.sub.2 (30.0 ml) was added DMSO (6.6 ml, 0.51 mmol) at
-78.degree. C. After 5 minutes, to the mixture was added
1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinemeth-
anol (1.2 g, 3.0 mmol) in CH.sub.2Cl.sub.2 (5.0 ml). The mixture
was stirred for 30 minutes at -78.degree. C., and triethylamine
(2.1 ml, 15.0 mmol) was added. The mixture was stirred for 30
minutes at -78.degree. C., and stirred for 30 minutes at room
temperature. Water was added to the mixture, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with water, then dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product, benzyl
cis-4-aminocyclohexanecarboxylate (769 mg, 3.3 mmol), and AcOH
(0.32 ml) in DCE (10 ml) was added NaBH(OAc).sub.3 (1.1 g, 5.4
mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by column chromatography on silica gel with
MeOH--CH.sub.2Cl.sub.2 (1:9, v/v) as eluent to give benzyl
cis-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrroli-
dinyl]methylamino]cyclohexanecarboxylate (1.5 g, 83%) as an
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.40-1.65 (m,
6H), 1.80-1.98 (m, 6H), 2.26 (s, 3H), 2.45-2.65 (m, 3H), 2.81-2.86
(m, 1H), 3.44-3.46 (m, 2H), 3.56 (s, 2H), 3.67 (s, 3H), 3.90-4.15
(m, 1H), 5.09 and 5.11 (each s, total 2H), 6.74-6.83 (m, 3H),
7.07-7.20 (m, 4H), 7.31-7.35 (m, 5H), 7.53-7.55 (m, 1H), 8.02-8.06
(m, 1H).
[1038] To a stirred solution of benzyl
cis-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-2-pyrrolidinyl]methylamino]cyclohexanecarboxylate (1.5 g,
2.45 mmol) in THF (10.0 ml) and MeOH (5.0 ml) was added 1N NaOH
(3.68 ml, 3.68 mmol). The mixture was stirred at 70.degree. C. for
18 hr. The mixture was concentrated in vacuo, water was added
thereto, and neutralized with 1N HCl. The mixture was concentrated
in vacuo. The residue was purified by column chromatography on
silica gel with MeOH--CH.sub.2Cl.sub.2 (1:5, v/v) as eluent to give
cis-4-[[1-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]-2-pyrrolidinyl]methylamino]cyclohexanecarboxy-
lic acid 171 (940 mg, 73%) as an amorphous solid. MW 522.64 IR
(KBr) 3283, 2945, 2860, 1534, 1453, 1415 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.38-2.00 (m, 12H), 2.45 (s, 3H), 2.30-3.95
(m, 4H), 3.22-3.75 (m, 2H), 3.58 (s, 2H), 3.86 (s, 3H), 4.12 (m,
1H), 6.73-7.16 (m, 5H), 7.77-7.79 (m, 1H), 7.98-8.02 (m, 1H),
8.51-8.52 (, 1H), 8.57-8.59 (m, 1H); MS (FAB) m/z 523 (M.sup.++1);
Anal. calcd for C.sub.29H.sub.38N.sub.4O.sub.5.0.5NaCl.2.2H.sub.2O:
C, 58.89; H, 7.23; N, 9.47. Found: C, 59.21; H, 7.11; N, 9.11.
Example 164
methyl
cis-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-p-
yrrolidinyl]methyl amino]cyclohexanecarboxylate HCl salt
[1039] ##STR1794##
[1040] SOCl.sub.2 was added to MeOH at 0.degree. C. After stirred
for 5 minutes,
cis-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]--
2-pyrrolidinyl]methylamino]cyclohexanecarboxylic acid (200 mg, 0.38
mmol) was added. The mixture was stirred at room temperature for 5
hr. The mixture was concentrated in vacuo. Aq. NaHCO.sub.3 was
added to the residue, and extracted with CH.sub.2Cl.sub.2. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with MeOH--CH.sub.2Cl.sub.2 (5:95 to
18:92, v/v) as eluent. The product was dissolved in EtOH (5.0 ml),
and 1N HCl (in EtOH) (1.0 ml, 1.0 mmol) was added thereto. The
mixture was concentrated in vacuo to give 172 (160 mg, 74%) as an
amorphous solid. MW 536.66 IR (KBr) 3247, 2950, 2875, 1731, 1671,
1612, 1533, 1454, 1205 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.45-2.10 (m, 12H), 2.25 (s, 3H), 2.60-2.70 (m, 1H),
2.90-3.20 (m, 3H), 2.50-2.55 (m, 2H), 3.63 (m, 5H), 3.86 (s, 3H),
4.15-4.30 (m, 1H), 6.74-7.16 (m, 5H), 7.76-7.78 (m, 1H), 8.00-8.09
(m, 1H), 8.54-8.70 (m, 2H); MS (FAB) m/z 537 (M.sup.++1); Anal.
calcd for C.sub.30H.sub.40N.sub.4O.sub.5.1.0HCl.1.0H.sub.2O: C,
60.95; H, 7.33; N, 9.48; Cl, 6.00. Found: C, 60.87; H, 7.47; N,
8.97: Cl, 5.90.
Example 165
4-[N-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidin-
yl]methyl]-N-methylamino]cyclohexanecarboxylic acid
[1041] ##STR1795##
[1042] To a stirred solution of methyl
cis-4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrroli-
dinyl]methylamino]cyclohexanecarboxylate (300 mg, 0.55 mmol), HCHO
(300 ml), and AcOH (66 mg, 11.1 mmol) in MeOH (10.0 ml) was added
NaBH.sub.3CN (70 mg, 1.1 mmol) at 0.degree. C. The reaction mixture
was stirred at room temperature for 18 hr. After concentrated in
vacuo, water was added and extracted with CH.sub.2Cl.sub.2. The
extract was washed with water, then dried over Na.sub.2SO.sub.4,
and concentrated in vacuo. The residue was purified by TLC with
MeOH--CH.sub.2Cl.sub.2 (3:97, v/v) as eluent to give methyl
4-[N-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nylmethyl]-N-methylamino]cyclohexanecarboxylate (160 mg, 52%) as an
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.30-2.20 (m,
12H), 2.27-2.37 (m, 6H), 2.50-2.60 (m, 1H), 3.30-3.80 (m, 5H), 3.55
(s, 2H), 3.66-3.73 (m, 6H), 4.10-4.20 (m, 1H), 6.60-7.55 (m, 7H),
8.03 (m, 1H), 8.15 (m, 1H).
[1043] To a stirred solution of methyl
4-[N-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidi-
nylmethyl]-N-methylamino]cyclohexanecarboxylate (100 mg, 0.18 mmol)
in THF (5.0 ml) and MeOH (2.5 ml) was added 1N NaOH (0.36 ml, 0.36
mmol). The mixture was stirred at 60.degree. C. for 18 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The mixture was concentrated in vacuo. The
residue was purified by TLC with MeOH--CH.sub.2Cl.sub.2 (1/4, v/v)
as eluent to give 173 (10 mg, 10%) as an amorphous solid. MW 536.66
IR (KBr) 3440, 2954, 1697, 1533, 1454 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.20-2.30 (m, 13H), 2.24 (s, 3H), 2.35-4.00
(m, 13H), 6.50-8.10 (m, 8H), 8.50 (m, 1H); MS (FAB) m/z 537
(M.sup.++1); Anal. calcd for
C.sub.30H.sub.40N.sub.4O.sub.5.2.0NaCl.0.8H.sub.2O: C, 53.94; H,
6.28; N, 8.39. Found: C, 54.08; H, 6.52; N, 8.04.
Example 166
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidi-
nyl]methoxy]cyclohexanecarboxylic acid
[1044] ##STR1796##
[1045] A mixture of methyl
4-(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxybenzoate (1.0 g,
2.9 mmol) and 5% Rh on alumina (500 mg) in EtOH (10.0 ml) and AcOH
(1.0 ml) was hydrogenated at room temperature at 5 atm for 36 hr.
The catalyst was filtered off, and the filtrate was concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (6:1, v/v) as eluent to give methyl
cis-4-[(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxy]cyclohexanecarbox-
ylate (900 mg, 89%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.46 (s, 9H), 1.46-2.00 (m, 12H), 2.34 (m, 1H), 3.20-3.55
(m, 5H), 3.67 (s, 3H), 3.84-3.92 (m, 1H).
[1046] To a stirred solution of methyl
cis-4-[(1-tert-butoxycarbonyl-(2S)-pyrrolidinyl)methoxy]cyclohexanecarbox-
ylate (900 mg, 2.6 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was added TFA
(5.0 ml) at 0.degree. C. The reaction mixture was stirred at room
temperature for 0.5 hr. The mixture was concentrated in vacuo. Sat.
NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (200 mg, 0.83 mmol),
3-methoxy-4-[N'-(2-methylphenyl)uredio]phenylacetic acid (260 mg,
0.83 mmol), HOBt (135 mg, 1.0 mmol), and triethylamine (344 .mu.l,
1.35-2.10 (m, 12H), 2.15-2.38 (m, 1H), 2.29 (m, 3H), 3.20-3.55 (m,
5H), 3.58 (s, 2H), 3.66 (s, 3H), 3.73 (s, 3H), 4.20-4.25 (m, 1H),
6.26-6.30 (m, 1H), 6.78-6.81 (m, 2H), 7.06-7.23 (m, 3H), 7.51-7.52
(m, 1H), 8.01-8.03 (m, 1H).
[1047] To a stirred solution of methyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolid-
inyl]methoxy]cyclohexanecarboxylate (460 mg, 0.86 mmol) in THF
(10.0 ml) and EtOH (5.0 ml) was added 1N NaOH (1.4 ml, 1.4 mmol).
The mixture was stirred at 60.degree. C. for 18 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 174 (370 mg, 83%) as a white crystalline
solid. MW 523.63 mp 110-113.degree. C.; IR (KBr) 3345, 2937, 1612,
1533, 1454 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.00-2.00
(m, 12H), 2.24 (s, 3H), 2.20-2.30 (m, 1H), 3.20-3.80 (m, 5H), 3.55
(s, 2H), 3.85 (s, 3H), 4.00-4.18 (m, 1H), 6.71-7.16 (m, 5H), 7.78
(d, J=8.0 Hz, 1H), 7.90 (d, J=8.3 Hz, 1H), 8.45 (s, 1H), 8.54 (s,
1H); MS (FAB) m/z 524 (M.sup.++1); Anal. calcd for
C.sub.29H.sub.37N.sub.3O.sub.6.0.2H.sub.2O: C, 66.07; H, 7.15; N,
7.97. Found: C, 66.02; H, 7.14; N, 7.87.
Example 167
4-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolid-
inyl]methoxy]cyclohexanecarboxylic acid
[1048] ##STR1797##
[1049] To a stirred solution of methyl
4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]cyclohexane
carboxylate (900 mg, 2.6 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was
added TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product (200 mg, 0.83 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (277 mg,
0.83 mmol), HOBt (135 mg, 1.0 mmol), and triethylamine (344 ml,
2.48 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl
(238 mg, 1.24 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 16 hr, and concentrated in vacuo.
Water was added to the residue, and extracted with EtOAc. The
extract was washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1:6, v/v) as eluent to give methyl
4-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrroli-
dinyl]methoxy]cyclohexane carboxylate (450 mg, 97%) as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.35-2.15 (m, 12H), 2.25-2.40
(m, 1H), 3.40-3.70 (m, 5H), 3.61 (s, 2H), 3.66 (s, 3H), 3.81 (s,
3H), 4.20-4.30 (m, 1H), 6.81-6.99 (m, 3H), 7.17-7.34 (m, 3H),
7.92-7.94 (m, 2H), 8.17-8.19 (m, 2H).
[1050] To a stirred solution of methyl
4-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrroli-
dinyl]methoxy]cyclohexanecarboxylate (450 mg, 0.86 mmol) in THF
(10.0 ml) and MeOH (5.0 ml) was added 1N NaOH (1.4 ml, 1.4 mmol).
The mixture was stirred at 70.degree. C. for 24 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 175 (370 mg, 84%) as a white crystalline
solid. MW 544.04 mp 111-115.degree. C.; IR (KBr) 3330, 2938, 1704,
1594, 1533, 1438, 1199 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6)
.delta. 1.00-2.00 (m, 12H), 2.20-2.30 (m, 1H), 3.20-3.80 (m, 5H),
3.55 (s, 2H), 3.85 (s, 3H), 4.00-4.20 (m, 1H), 6.73-6.75 (m, 1H),
6.87 (s, 1H), 6.99-7.03 (m, 1H), 7.25-7.29 (m, 1H), 7.42 (d, J=7.1
Hz, 1H), 7.95 (d, J=8.3 Hz, 1H), 8.08 (d, J=8.0, 1H), 8.78 (s, 1H),
8.92 (s, 1H); MS (FAB) m/z 544 (M.sup.++1); Anal calcd for
C.sub.28H.sub.34N.sub.3O.sub.6Cl.0.2H.sub.2O: C, 61.41; H, 6.33; N,
7.67; Cl, 6.47. Found: C, 61.37; H, 6.32; N, 7.56; Cl, 6.55.
Example 168
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylactyl]-(2S)-pyrrolidinyl-
]methoxy cyclohexanecarboxylic acid
[1051] ##STR1798##
[1052] To a stirred solution of methyl
4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]cyclohexane
carboxylate (900 mg, 2.6 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was
added TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product (200 mg, 0.83 mmol),
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (314 mg,
0.83 mmol), HOBt (135 mg, 1.0 mmol), and triethylamine (344 ml,
2.48 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl
(238 mg, 1.24 mmol) at 0.degree. C. The reaction mixture was
stirred at room temperature for 16 hr, and concentrated in vacuo.
Water was added to the residue, and extracted with EtOAc. The
extract was washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1:6, v/v) as eluent to give methyl
[4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolid-
inyl]methoxy]cyclohexane carboxylate (450 mg, 90%) as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.30-2.10 (m, 12H), 2.35-2.40
(m, 1H), 3.25-3.70 (m, 5H), 3.84 (s, 3H), 4.10-4.25 (m, 1H),
6.81-7.06 (m, 4H), 7.25-7.32 (m, 2H), 7.50-7.52 (m, 1H), 7.90-7.92
(m, 1H), 8.13-8.15 (m, 1H).
[1053] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-pyrrolidi-
nyl]methoxycyclohexanecarboxylate (450 mg, 0.74 mmol) in THF (10.0
ml) and MeOH (5.0 ml) was added 1N NaOH (1.2 ml, 1.2 mmol). The
mixture was stirred at 70.degree. C. for 24 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 176 (340 mg, 77%) as a white crystalline
solid. MW 588.49 mp 108-111.degree. C.; IR (KBr) 3328, 2938, 1702,
1594, 1529, 1434 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.00-2.00 (m, 12H), 2.15-2.25 (m, 1H), 3.40-3.75 (m, 5H), 3.48 (s,
2H), 3.85 (s, 3H), 4.04-4.15 (m, 1H), 6.70-6.72 (m, 1H), 6.87 (s,
1H), 6.94-6.98 (m, 1H), 7.29-7.33 (m, 1H), 7.59 (d, J=8.1 Hz, 1H),
7.93-7.95 (m, 2H), 8.73-8.74 (m, 1H), 8.91-8.92 (m, 1H); MS (FAB)
m/z 589 (M.sup.++1); Anal. calcd for
C.sub.28H.sub.34N.sub.3O.sub.6Br.0.2H.sub.2O: C, 56.80; H, 5.86; N,
7.10; Br, 13.49. Found: C, 56.66; H, 5.83; N, 6.97; Br, 13.66.
Example 169
4-[[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidinyl]methox-
y]cyclohexane carboxylic acid
[1054] ##STR1799##
[1055] To a stirred solution of methyl
4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl)methoxy]cyclohexane
carboxylate (450 mg, 1.3 mmol) in CH.sub.2Cl.sub.2 (5.0 ml) was
added TFA (5.0 ml) at 0.degree. C. The reaction mixture was stirred
at room temperature for 0.5 hr. The mixture was concentrated in
vacuo. Sat. NaHCO.sub.3 was added to the residue, and extracted
with CH.sub.2Cl.sub.2. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product
was used to the subsequent reaction without further purification.
To a stirred solution of the crude product,
4-[N'-(2-methylphenyl)uredio]phenylacetic acid (375 mg, 1.3 mmol),
HOBt (178 mg, 1.3 mmol), and triethylamine (550 ml, 3.9 mmol) in
THF (6.0 ml) and MeCN (6.0 ml) was added EDC.HCl (380 mg, 1.9 mmol)
at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:6, v/v) as eluent to give methyl
4-[[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidinyl]metho-
xy]cyclohexanecarboxylate (520 mg, 78%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.30-2.40 (m, 13H), 2.21 (s, 3H),
3.30-3.80 (m, 7H), 3.65 (s, 3H), 4.10-4.30 (m, 1H), 6.90-7.20 (m,
8H), 7.40-7.70 (m, 2H).
[1056] To a stirred solution of methyl
4-[[1-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S)-pyrrolidinyl]metho-
xy]cyclohexanecarboxylate (520 mg, 1.0 mmol) in THF (10.0 ml) and
MeOH (5.0 ml) was added 1N NaOH (1.5 ml, 1.5 mmol). The mixture was
stirred at 70.degree. C. for 24 hr. The mixture was concentrated in
vacuo, water was added thereto, and neutralized with 1N HCl. The
resulting solid was collected, washed with water, and dried in
vacuo to give 177 (450 mg, 91%) as a white crystalline solid. MW
493.60 mp 107-111.degree. C.; IR (KBr) 3353, 2938, 1704, 1540,
1454, 1240 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.20-2.00
(m, 12H), 2.23 (s, 3H), 2.22-2.24 (m, 1H), 3.20-3.80 (m, 7H),
4.00-4.18 (m, 1H), 6.90-6.94 (m, 1H), 7.10-7.16 (m, 5H), 7.36-7.38
(m, 2H), 7.82-7.87 (m, 2H), 8.89 (s, 1H), 12.0 (br s, 1H); MS (FAB)
m/z 494 (M.sup.++1); Anal. calcd for
C.sub.28H.sub.35N.sub.3O.sub.5.0.2H.sub.2O: C, 67.64; H, 7.18; N,
8.45. Found: C, 67.66; H, 7.19; N, 8.24.
Example 170
cis-4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-octah-
ydroindolylmethoxy]cyclohexanecarboxylic acid
[1057] ##STR1800##
[1058] To a stirred solution of
[1-tert-butoxycarbonyl-(2S)-octahydroindolyl]carboxylic acid (1.0
g, 3.7 mmol) in THF (10.0 ml) was added BH.sub.3-THF (1.0 M in THF,
8.0 ml) at 0.degree. C. After stirred at room temperature for 1.0
h, the reaction mixture was heated under reflux for 1.5 hr. After
cooled, the mixture was concentrated in vacuo. Water was added
thereto at 0.degree. C., and extracted with EtOAc. The extract was
washed with water, then dried over Na.sub.2SO.sub.4, and
concentrated in vacuo to give
[1-tert-butoxycarbonyl-(2S)-octahydroindolyl]methanol (947 mg,
quant) as a colorless oil.
[1059] To a stirred solution of
[1-tert-butoxycarbonyl-(2S)-octahydroindolyl]methanol (947 mg, 3.7
mmol), methyl 4-hydroxybenzoate (565 mg, 3.7 mmol), and Ph.sub.3P
(1.2 g, 4.5 mmol) in THF (10 ml) was added DIAD (984 mg, 4.5 mmol)
at 0.degree. C. The reaction mixture was stirred at room
temperature for 18 hr. The mixture was concentrated in vacuo. The
residue was purified by column chromatography on silica gel with
n-hexane-EtOAc (9/1, v/v) as eluent to give methyl
4-[1-tert-butoxycarbonyl-(2S)-octahydroindolylmethoxy]benzoate (700
mg, 50%) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.10-2.25 (m, 1H), 1.45 (s, 9H), 3.88 (s, 3H), 3.70-4.20 (n, 3H),
4.36 (br s, 1H), 6.94 (d, J=8.8 Hz, 2H), 7.96 (d, J=8.5 Hz,
2H).
[1060] A mixture of methyl
4-[1-tert-butoxycarbonyl-(2S)-octahydroindolylmethoxy]benzoate (700
mg, 1.8 mmol) and 5% Rh on alumina (400 mg) in EtOH (10.0 ml) and
AcOH (1.0 ml) was hydrogenated at room temperature at 5 atm for 48
hr. The catalyst was filtered off, and the filtrate was
concentrated in vacuo. The residue was purified by column
chromatography on silica gel with n-hexane-EtOAc (7:1, v/v) as
eluent to give methyl
cis-4-[1-tert-butoxycarbonyl-(2S)-octahydroindolylmethoxy]cyclohexanecarb-
oxylate (600 mg, 85%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.10-2.35 (m, 20H), 1.44 (s, 9H), 3.45-3.90
(m, 5H), 3.80 (s, 3H).
[1061] To a stirred solution of methyl
cis-4-[1-tert-butoxycarbonyl-(2S)-octahydroindolylmethoxy]cyclohexanecarb-
oxylate (600 mg, 1.5 mmol) in CH.sub.2Cl.sub.2 (6.0 ml) was added
TFA (6.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (221 mg, 0.75 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (250 mg,
0.75 mmol), HOBt (101 mg, 0.75 mmol), and triethylamine (312 ml,
2.3 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl
(216 mg, 1.1 mmol) at 0.degree. C. The reaction mix was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The residue was purified by column chromatography on silica
gel with n-hexane-EtOAc (1:3, v/v) as eluent to give methyl
cis-4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-octa-
hydroindolylmethoxy]cyclohexane carboxylate (430 mg, 94%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.10-2.40 (m, 20H),
3.45 (br s, 1H), 3.62 (s, 2H), 3.66 (s, 3H), 3.73 (s, 3H),
3.60-3.85 (m, 2H), 4.09-4.14 (m, 2H), 6.75-6.98 (m, 3H), 7.22-2.46
(m, 4H), 7.92 (d, J=8.0 Hz, 1H), 8.18 (d, J=8.3 Hz, 1H).
[1062] To a stirred solution of methyl
cis-4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-octa-
hydroindolylmethoxy]cyclohexanecarboxylate (430 mg, 0.7 mmol) in
THF (10.0 ml) and MeOH (5.0 ml) was added 1N NaOH (1.4 ml, 1.4
mmol). The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 178 (360 mg, 86%) as a white
crystalline solid. MW 598.13 mp 120-121.degree. C.; IR (KBr) 3338,
2933, 2859, 1614, 1533, 1438 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3)
.delta. 1.05-2.40 (m, 20H), 3.38-3.50 (m, 2H), 3.63 and 3.65 (each
s, total 2H), 3.71 and 3.75 (each s, total 3H), 3.70-3.80 (m, 1H),
3.93-3.97 (m, 1H), 4.15 (br s, 1H), 6.75-6.77 (m, 2H), 6.93-6.97
(m, 1H), 7.20-7.32 (m, 3H), 7.60-7.63 (m, 1H), 7.85 (d, J=8.3 Hz,
1H), 8.16 (d, J=8.3 Hz, 1H); MS (FAB) m/z 598 (M.sup.++1); Anal.
calcd for C.sub.32H.sub.40N.sub.3O.sub.6Cl.0.5H.sub.2O: C, 63.30;
H, 6.81; N, 6.92; Cl, 5.84. Found: C, 63.68; H, 6.81; N, 6.81; Cl,
5.98.
Example 171
cis-4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-octahy-
droindolylmethoxy]cyclohexanecarboxylic acid
[1063] ##STR1801##
[1064] To a stirred solution of methyl
cis-4-[1-tert-butoxycarbonyl-(2S)-octahydroindolylmethoxy]cyclohexanecarb-
oxylate (600 mg, 1.5 mmol) in CH.sub.2Cl.sub.2 (6.0 ml) was added
TFA (6.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (221 mg, 0.75 mmol),
4-[N'-(2-bromophenyl)uredio]-3-methoxyphenylacetic acid (284 mg,
0.75 mmol), HOBt (101 mg, 0.75 mmol), and triethylamine (312 ml,
2.3 mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl
(216 mg, 1.1 mmol) at 0.degree. C. The reaction mixture was stirred
at room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2 M citric acid, and sat NaHCO.sub.3,
then dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The
residue was purified by column chromatography on silica gel with
n-hexane-EtOAc (1:3, v/v) as eluent to give methyl
cis-4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-octah-
ydroindolylmethoxy]cyclohexane carboxylate (480 mg, 96%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.10-2.40 (m, 20H),
3.45 (br s, 1H), 3.61 (s, 2H), 3.66 (s, 3H), 3.76 (s, 3H),
3.60-3.80 (m, 2H), 4.11-4.14 (m, 2H), 6.76-6.92 (m, 3H), 7.25-7.32
(m, 3H), 7.49 (d, J=7.1 Hz, 1H), 7.90 (d, J=8.0 Hz, 1H), 8.13 (d,
J=8.0 Hz, 1H).
[1065] To a stirred solution of methyl
cis-4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxy]phenylacetyl]-(2S)-octah-
ydroindolylmethoxy]cyclohexanecarboxylate (480 mg, 0.73 mmol) in
THF (10.0 ml) and MeOH (5.0 ml) was added 1N NaOH (1.5 ml, 1.5
mmol). The mixture was stirred at 70.degree. C. for 24 hr. The
mixture was concentrated in vacuo, water was added thereto, and
neutralized with 1N HCl. The resulting solid was collected, washed
with water, and dried in vacuo to give 179 (400 mg, 85%) as a white
crystalline solid. MW 642.58 mp 115-120.degree. C.; IR (KBr) 3332,
2933, 2859, 1704, 1592, 1529, 1434 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.10-2.40 (m, 20H), 3.40-3.50 (m, 2H), 3.61
and 3.63 (each s, total 2H), 3.75 and 3.78 (each s, total 3H),
3.70-3.80 (m, 1H), 3.90-3.93 (m, 1H), 4.15 (br s, m), 6.76-6.92 (m,
3H), 7.26-7.30 (m, 1H), 7.43-7.52 (m, 3H), 7.84-7.86 (m, 1H),
8.10-8.12 (m, 1H); MS (FAB) m/z 643 (M.sup.++1); Anal. calcd for
C.sub.32H.sub.40N.sub.3O.sub.6Br.0.4H.sub.2O: C, 59.15; H, 6.33; N,
6.49; Br, 12.30. Found: C, 59.26; H, 6.33; N, 6.36; Br, 12.37.
Example 172
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidinyl-
]carbonylamino]cyclohexanecarboxylic acid
[1066] ##STR1802##
[1067] To a stirred solution of benzyl
4-aminocyclohexanecarboxylate (900 mg, 3.9 mmol), boc-proline (830
mg, 3.9 mmol), HOBt (521 mg, 3.9 mmol), and triethylamine (1.6 ml,
11.6 mmol) in CH.sub.2Cl.sub.2 (30.0 ml) was added EDC.HCl (1.1 g,
5.8 mmol) at 0.degree. C. The reaction mixture was stirred at room
temperature for 16 hr, and concentrated in vacuo. Water was added
to the residue, and extracted with EtOAc. The extract was washed
with sat. NaHCO.sub.3, 2-M citric acid, and sat. NaHCO.sub.3, then
dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The residue
was purified by column chromatography on silica gel with
n-hexane-EtOAc (6:1, v/v) as eluent to give benzyl
cis-4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl)carbonylamino]cyclohexanecar-
boxylate (600 mg, 36%) as an amorphous solid. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.44 (s, 9H), 1.50-1.90 (m, 12H), 2.20-2.26
(m, 1H), 3.25-3.50 (m, 2H), 3.80-3.90 (m, 1H), 4.10-4.25 (m, 1H),
5.12 (s, 2H), 7.35-7.36 (m, 5H).
[1068] To a stirred solution of benzyl
cis-4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl)carbonylamino]cyclohexanecar-
boxylate (600 mg, 1.4 mmol) in CH.sub.2Cl.sub.2 (6.0 ml) was added
TFA (3.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (300 mg, 0.7 mmol),
3-methoxy-4-[N'-(2-methylphenyl)uredio]phenylacetic acid (220 mg,
0.7 mmol), HOBt (94 mg, 0.7 mmol), and triethylamine (291 ml, 2.1
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (201
mg, 1.1 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2-M citric acid, and sat NaHCO.sub.3,
then dried over Na.sub.2SO.sub.4, and concentrated in vacuo. The
resulting solid was collected and washed with EtOAc to give benzyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido)phenylacetyl]-2-pyrrolidiny-
l]carbonylamino]cyclohexanecarboxylate (380 mg, 87%) as a white
crystalline solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.40-2.15 (m,
12H), 2.28 (m, 3H), 2.30-2.50 (m, 2H), 3.40-3.55 (m, 2H), 3.61 (s,
2H), 3.71 (s, 3H), 3.82 (m, 1H), 4.53 (d, J=6.3 Hz, 1H), 5.10 (s,
2H), 6.42 (s, 1H), 6.77-6.79 (m, 2H), 7.04-7.34 (m, 9H), 7.50-7.52
(m, 1H), 8.05-8.07 (m, 1H).
[1069] To a stirred solution of benzyl
4-[[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-pyrrolidiny-
l]carbonylamino]cyclohexanecarboxylate (380 mg, 0.6 mmol) in THF
(10.0 ml) and EtOH (5.0 ml) was added 1N NaOH (0.9 ml, 0.9 mmol).
The mixture was stirred at 50.degree. C. for 18 hr. The mixture was
concentrated in vacuo, water was added thereto, and neutralized
with 1N HCl. The resulting solid was collected, washed with water,
and dried in vacuo to give 180 (230 mg, 71%) as a white crystalline
solid. MW 636.62 mp 136-142.degree. C.; IR (KBr) 3345, 2940, 1650,
1625, 1535, 1454 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta.
1.40-2.00 (m, 12H), 2.24 (s, 3H), 2.30-2.40 (m, 1H), 3.45-3.80 (m,
5H), 3.86-3.87 (m, 3H), 4.30-4.43 (m, 1H), 6.65-7.30 (m, 5H),
7.70-7.80 (m, 1H), 7.98-8.09 (m, 1H), 8.46-8.57 (m, 1H); MS (FAB)
m/z 537 (M.sup.++1); Anal. calcd for
C.sub.29H.sub.36N.sub.4O.sub.6.0.5H.sub.2O: C, 63.84; H, 6.83; N,
10.27. Found: C, 64.18; H, 6.91; N, 9.85.
Example 173
4-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-2-pyrrolidinyl-
]carbonylamino]cyclohexanecarboxylic acid
[1070] ##STR1803##
[1071] To a stirred solution of benzyl
cis-4-[(1-tert-butoxycarbonyl-2-pyrrolidinyl)carbonylamino]cyclohexanecar-
boxylate (600 mg, 1.4 mmol) in CH.sub.2Cl.sub.2 (6.0 ml) was added
TFA (3.0 ml) at 0.degree. C. The reaction mixture was stirred at
room temperature for 0.5 hr. The mixture was concentrated in vacuo.
Sat. NaHCO.sub.3 was added to the residue, and extracted with
CH.sub.2Cl.sub.2. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The crude product was
used to the subsequent reaction without further purification. To a
stirred solution of the crude product (300 mg, 0.7 mmol),
4-[N'-(2-chlorophenyl)uredio]-3-methoxyphenylacetic acid (237 mg,
0.7 mmol), HOBt (94 mg, 0.7 mmol), and triethylamine (291 ml, 2.1
mmol) in THF (10.0 ml) and MeCN (10.0 ml) was added EDC.HCl (201
mg, 1.1 mmol) at 0.degree. C. The reaction mixture was stirred at
room temperature for 16 hr, and concentrated in vacuo. Water was
added to the residue, and extracted with EtOAc. The extract was
washed with sat. NaHCO.sub.3, 2-M citric acid, and sat.
NaHCO.sub.3, then dried over Na.sub.2SO.sub.4, and concentrated in
vacuo. The resulting solid was collected and washed with EtOAc to
give benzyl
4-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenylacetyl]-2-pyrrolidin-
yl]carbonylamino]cyclohexanecarboxylate (310 mg, 68%) as a white
crystalline solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.40-2.15 (m,
12H), 2.30-2.60 (m, 2H), 3.42-3.55 (m, 2H), 3.64 (s, 2H), 3.84 (s,
3H), 4.55 (d, J=6.1 Hz, 1H), 5.12 (s, 2H), 6.81-70.35 (m, 12H),
7.96-7.98 (m, 1H), 8.17-8.19 (m, 1H).
[1072] To a stirred solution of methyl benzyl
4-[[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy]phenyl
acetyl]-2-pyrrolidinyl]carbonylamino]cyclohexanecarboxylate (310
mg, 0.86 mmol) in THF (10.0 ml) and MeOH (5.0 ml) was added 1N NaOH
(0.7 ml, 0.7 mmol). The mixture was stirred at 50.degree. C. for 18
hr. The mixture was concentrated in vacuo, water was added thereto,
and neutralized with 1N HCl. The resulting solid was collected,
washed with water, and dried in vacuo to give 181 (260 mg, 98%) as
a white crystalline solid. MW 557.03 mp 135-140.degree. C.; IR
(KBr) 3328, 2938, 1594, 1533, 1438, 1203 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.40-2.20 (m, 12H), 2.30-2.40 (m, 1H),
3.40-3.80 (m, 5H), 3.65-3.85 (m, 3H), 4.30-4.43 (m, 1H), 6.66-7.31
(m, 5H), 7.42-8.10 (m, 2H), 8.89-8.94 (m, 2H); MS (FAB) m/z 557
(M.sup.++1); Anal. calcd for
C.sub.28H.sub.33N.sub.4O.sub.6Cl.3H.sub.2O: C, 59.79; H, 6.02; N,
9.96; Cl, 6.30. Found: C, 59.86; H, 6.10; N, 9.60; Cl, 6.34.
Example 174
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(3S)-pyrrolidin-
yloxy]benzoic acid
[1073] ##STR1804##
[1074] To a cooled (0.degree. C.), stirred solution of methyl
4-hydroxybenzoate (1.84 g, 12.1 mmol), (R)-1-benzyl-3-pyrrolidinol
(2.00 ml, 12.1 mmol), and Ph.sub.3P (3.81 g, 14.5 mmol) in THF (25
ml) was added DIAD (2.86 ml, 14.5 mmol) and the reaction mixture
was heated under reflux for 10 hr. After cooled to room
temperature, the mixture was evaporated. The residue was purified
by column chromatography on silica-gel with n-hexane-EtOAc (3:1,
v/v) as eluent to give methyl
4-[1-benzyl-(3S)-pyrrolidinyloxy]benzoate (3.66 g, 97%) as a pale
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.95-2.02 (m, 1H),
2.28-2.37 (m, 1H), 2.57-2.63 (m, 1H), 2.72-2.81 (m, 2H), 2.96-3.01
(m, 1H), 3.63-3.71 (m, 2H), 3.87 (s, 3H), 4.84-4.89 (m, 1H), 6.84
(d, J=8.8 Hz, 2H), 7.23-7.34 (m, 5H), 7.95 (d, J=8.8 Hz, 2H).
[1075] A solution of methyl
4-[1-benzyl-(3S)-pyrrolidinyloxy]benzoate (3.66 g, 11.8 mmol) in
MeOH (25 ml) was hydrogenated over Pd(OH).sub.2/C (0.73 g, 20 wt %)
overnight. The reaction mixture was filtered to remove the catalyst
and the solution was evaporated to give methyl
4-[(3S)-pyrrolidinyloxy]benzoate (2.60 g, q.y.) as a pale yellow
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.94-2.01 (m, 2H), 2.09-2.18
(m, 1H), 2.91-2.97 (m, 1H), 3.04-3.09 (m, 1H), 3.16-3.23 (m, 2H),
3.88 (s, 3H), 4.88-4.91 (m, 1H), 6.86 (m, 2H), 7.96-7.98 (m, 2H);
MS (ESI) m/z 222 (M.sup.++1).
[1076] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (449 mg,
1.43 mmol), methyl 4-[(3S)-pyrrolidinyloxy]benzoate (316 mg, 1.43
mmol), EDC.HCl (330 mg, 1.72 mmol), HOBt (193 mg, 1.43 mmol), and
Et.sub.3N (240 ml, 1.72 mmol) in THF (5 ml) was stirred at room
temperature overnight The reaction mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(3S)-pyrrolidi-
nyloxy]benzoate (735 mg, 99%) as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.03-2.31 (series of s and m, total 5H),
3.57-3.78 (series of m, total 9H), 3.88 (s, 3H), 4.95-4.99 (m, 1H),
6.73-7.00 (m, 5H), 7.06-7.10 (m, 1H), 7.18-7.22 (m, 2H), 7.42-7.46
(m, 1H), 7.57-7.62 (m, 1H), 7.95-8.08 (m, 3H); MS (ESI) m/z 518
(M.sup.++1).
[1077] To a stirred solution of methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(3S)-pyrrolidi-
nyloxy]benzoate (627 mg, 1.21 mmol) in THF (5 ml) was added 0.5 N
NaOH (5 ml) and the reaction mixture was heated under reflux for 3
hr. After cooled to room temperature, the mixture was poured into
ice-1 N HCl and the resulting precipitate was collected under a
reduced pressure. The crude solid was recrystallized from
MeOH--CHCl.sub.3--IPE to give 182 (235 mg, 39%) as a white
crystalline powder. MW 503.55 mp 131-135.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.06-2.27 (series of m, total 5H), 3.57-3.64
(m, 4H), 3.71-3.88 (series of s and m, total 5H), 5.11 and 5.20
(each m, total 1H), 6.73-6.77 (m, 1H), 6.88-6.95 (m, 2H), 7.02-7.05
(m, 2H), 7.11-7.17 (m, 2H), 7.79-7.81 (m, 1H), 7.88-7.90 (m, 2H),
7.98-8.03 (m, 1H), 8.45-8.47 (m, 1H), 8.55-8.57 (m, 1H), 12.66 (br
s, 1H); MS (ESI) m/z 504 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.29N.sub.3O.sub.6.3/4H.sub.2O: C, 65.04; H, 5.95; N,
8.13. Found: C, 65.11; H, 5.99; N, 7.66.
Example 175
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(3S)-pyrrolidin-
yloxy]benzoic acid
[1078] ##STR1805##
[1079] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (410 mg,
1.22 mmol), methyl 4-[(3S)-pyrrolidinyloxy]benzoate (270 mg, 1.22
mmol), EDC.HCl (280 mg, 1.46 mmol), HOBt (200 mg, 1.48 mmol), and
Et.sub.3N (205 ml, 1.47 mmol) in THF (8 ml) was stirred at room
temperature overnight. The reaction mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 60:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(3S)-pyrrolidi-
nyloxy]benzoate (652 mg, 99%) as a white solid. mp 200-203.degree.
C.; .sup.1H-NMR (CDCl.sub.3) .delta. 2.06-2.32 (m, 2H), 3.60-3.82
(series of m, total 9H), 3.88 (s, 3H), 4.97-5.01 (m, 1H), 6.76-6.86
(m, 4H), 6.95-6.99 (m, 1H), 7.23-7.47 (m, 4H), 7.91-7.99 (m, 3H),
8.19-8.21 (m, 1H); MS (ESI) m/z 537 (M.sup.+).
[1080] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(3S)-pyrrolidi-
nyloxy]benzoate (650 mg, 1.21 mmol) in THF (5 ml) was added 0.5 N
NaOH (5 ml) and the reaction mixture was heated under reflux for 5
hr. After cooled to room temperature, the mixture was poured into
ice-1 N HCl and the resulting precipitate was collected under a
reduced pressure. The crude solid was recrystallized from
MeOH--CHCl.sub.3--IPE to give 183 (443 mg, 70%) as a pale yellow
crystalline powder. MW 523.97 mp 190-193.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.06-2.27 (m, 2H), 3.56-3.62 (m, 4H),
3.71-3.88 (series of s and m, total 5H), 5.11 and 5.20 (each m,
total 1H), 6.74-6.78 (m, 1H), 6.89-6.91 (m, 1H), 7.00-7.05 (m, 3H),
7.26-7.30 (m, 1H), 7.43-7.45 (m, 1H), 7.88-7.98 (m, 3H), 8.08-8.10
(m, 1H), 8.89-8.95 (m, 2H), 12.67 (br s, 1H); MS (ESI) m/z 524
(M.sup.++1); Anal. Calcd for
C.sub.27H.sub.26ClN.sub.3O.sub.6.1/4H.sub.2O: C, 61.36; H, 5.05; N,
7.95; Cl, 6.71. Found: C, 61.69; H, 5.45; N, 7.29; Cl, 6.91.
Example 176
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(3S)-pyrrolidiny-
loxy]benzoic acid
[1081] ##STR1806##
[1082] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (540 mg,
1.42 mmol), methyl 4-[(3S)-pyrrolidinyloxy]benzoate (315 mg, 1.42
mmol), EDC.HCl (328 mg, 1.71 mmol), HOBt (230 mg, 1.70 mmol), and
Et.sub.3N (240 ml, 1.72 mmol) in THF (8 ml) was stirred at room
temperature overnight. The reaction mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(3S)-pyrrolidin-
yloxy]benzoate (620 mg, 74%) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.06-2.33 (m, 2H), 3.60-3.82 (series of m,
total 9H), 3.89 (s, 3H), 4.97-5.01 (m, 1H), 6.77-7.00 (m, 5H),
7.27-7.40 (m, 3H), 7.49-7.51 (m, 1H), 7.91-7.99 (m, 3H), 8.13-8.17
(m, 1H); MS (ESI) m/z 583 (M.sup.++1).
[1083] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetyl]-(3S)-pyrrolidinyloxy]benzoate (620 mg, 1.06 mmol) in THF (5
ml) was added 0.5 N NaOH (5 ml) and the reaction mixture was heated
under reflux for 2.5 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl and the resulting precipitate
was collected under a reduced pressure. The crude solid was
recrystallized from MeOH--CHCl.sub.3--IPE to give 184 (421 mg, 70%)
as a white crystalline powder. MW 568.42 mp 173-175.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 2.06-2.28 (m, 2H), 3.56-3.65 (m,
4H), 3.71-3.88 (series of s and m, total 5H), 5.12 and 5.20 (each
m, total 1H), 6.74-6.78 (m, 1H), 6.89-7.05 (m, 4H), 7.31-7.34 (m,
1H), 7.59-7.61 (m, 1H), 7.88-7.98 (m, 4H), 8.73-8.74 (m, 1H),
8.91-8.93 (m, 1H), 12.67 (br s, 1H); MS (ESI) m/z 569 (M.sup.++1);
Anal. Calcd for C.sub.27H.sub.26BrN.sub.3O.sub.6: C, 57.05; H,
4.61; N, 7.39; Br, 14.06. Found: C, 57.57; H, 5.12; N, 6.81; Br,
13.96.
Example 177
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(3R)-pyrrolidin-
yloxy]benzoic acid
[1084] ##STR1807##
[1085] To a cooled (0.degree. C.), stirred solution of methyl
4-hydroxybenzoate (1.78 g, 11.7 mmol), 1-benzyl-(3S)-pyrrolidinol
(2.07 g, 11.7 mmol), and Ph.sub.3P (3.68 g, 14.0 mmol) in THF (25
ml) was added DIAD (2.76 ml, 14.0 mmol) and the reaction mixture
was heated under reflux for 10 hr. After cooled to room
temperature, the mixture was evaporated. The residue was purified
by column chromatography on silica-gel with n-hexane-EtOAc (3:1,
v/v) as eluent to give methyl
4-[1-benzyl-(3R)-pyrrolidinyloxy]benzoate (3.56 g, 98%) as a pale
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.95-2.03 (m, 1H),
2.28-2.37 (m, 1H), 2.57-2.63 (m, 1H), 2.73-2.81 (m, 2H), 2.97-3.01
(m, 1H), 3.63-3.72 (m, 2H), 3.87 (s, 3H), 4.85-4.90 (m, 1H),
6.83-6.85 (m, 2H), 7.27-7.34 (m, 5H), 7.94-7.97 (m, 2H); MS (ESI)
m/z 312 (M.sup.++1).
[1086] A solution of methyl
4-[1-benzyl-(3R)-pyrrolidinyloxy]benzoate (3.56 g, 11.4 mmol) in
MeOH (25 ml) was hydrogenated over Pd(OH).sub.2/C (0.72 g, 20 wt %)
overnight. The reaction mixture was filtered to remove the catalyst
and the solution was evaporated to give methyl
4-[(3R)-pyrrolidinyloxy]benzoate (2.53 g, q.y.) as a pale yellow
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.94-2.18 (m, 3H), 2.91-2.97
(m, 1H), 3.04-3.09 (m, 1H), 3.16-3.22 (m, 2H), 3.88 (s, 3H),
4.88-4.91 (m, 1H), 6.86-6.89 (m, 2H), 7.97-7.99 (m, 2H); MS (ESI)
m/z 263 [M.sup.++1+41, (+MeCN)].
[1087] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (460 mg,
1.46 mmol), methyl 4-[(3R)-pyrrolidinyloxy]benzoate (324 mg, 1.46
mmol), EDC.HCl (337 mg, 1.76 mmol), HOBt (237 mg, 1.75 mmol), and
Et.sub.3N (245 ml, 1.76 mmol) in THF (10 ml) was stirred at room
temperature overnight The reaction mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(3R)-pyrrolidi-
nyloxy]benzoate (583 mg, 77%) as a colorless oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.06-2.23 (m, 2H), 2.30 (s, 3H), 3.58-3.89
(series of s and m, total 12H), 4.95-4.99 (m, 1H), 6.75-7.00 (m,
4H), 7.13-7.30 (m, 5H), 7.52-7.57 (m, 1H), 7.96-8.03 (m, 3H); MS
(ESI) m/z 518 (M.sup.++1).
[1088] To a stirred solution of methyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(3R)-pyrrolidi-
nyloxy]benzoate (583 mg, 1.13 mmol) in THF (5 ml) was added 0.5 N
NaOH (5 ml) and the reaction mixture was heated under reflux for 3
hr. After cooled to room temperature, the mixture was poured into
ice-1 N HCl and the resulting precipitate was collected under a
reduced pressure. The crude solid was recrystallized from
MeOH--CHCl.sub.3-Et.sub.2O to give 185 (297 mg, 52%) as a white
crystalline powder. MW 503.55 mp 158-162.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.08-2.31 (series of s and m, total 5H),
3.54-3.89 (series of m, total 9H), 5.11 and 5.20 (each m, total
1H), 6.72-7.17 (series of m, total 6H), 7.78-7.80 (m, 1H),
7.87-7.90 (m, 2H), 7.98-8.02 (m, 2H), 8.46-8.47 (m, 1H), 8.55-8.57
(m, 1H), 12.66 (br s, 1H); MS (ESI) m/z 504 (M.sup.++1); Anal.
Calcd for C.sub.28H.sub.29N.sub.3O.sub.6.1/4H.sub.2O: C, 66.19; H,
5.85; N, 8.27. Found: C, 66.12; H, 5.77; N, 8.21.
Example 178
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(3R)-pyrrolidin-
yloxy]benzoic acid
[1089] ##STR1808##
[1090] A mixture of
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (498 mg,
1.49 mmol), methyl 4-[(3R)-pyrrolidinyloxy]benzoate (329 mg, 1.49
mmol), EDC.HCl (342 mg, 1.78 mmol), HOBt (241 mg, 1.78 mmol), and
Et.sub.3N (250 ml, 1.79 mmol) in THF (10 ml) was stirred at room
temperature overnight. The reaction mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 50:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetyl]-(3R)-pyrrolidi-
nyloxy]benzoate (561 mg, 70%) as a white form. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.05-2.34 (m, 2H), 3.59-4.07 (series of s and
m, total 12H), 4.97-5.02 (m, 1H), 6.75-6.86 (m, 4H), 6.94-7.00 (m,
1H), 7.22-7.33 (m, 2H), 7.59-7.66 (m, 2H), 7.92-7.99 (m, 3H),
8.19-8.22 (m, 1H); MS (ESI) m/z 538 (M.sup.++1).
[1091] To a stirred solution of methyl
4-[1-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetyl]-(3R)-pyrrolidinyloxy]benzoate (561 mg, 1.04 mmol) in THF (5
ml) was added 0.5 N NaOH (5 ml) and the reaction mixture was heated
under reflux for 5 hr. After cooled to room temperature, the
mixture was poured into ice-1 N HCl and the resulting precipitate
was collected under a reduced pressure. The crude solid was
recrystallized from MeOH--CHCl.sub.3-Et.sub.2O to give 186 (361 mg,
66%) as a white crystalline powder. MW 523.97 mp 193-194.degree.
C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.06-2.28 (m, 2H), 3.58-3.62
(m, 4H), 3.71-3.76 (m, 1H), 3.83-3.89 (series of s and m, total
4H), 5.12 and 5.20 (each m, total 1H), 6.74-6.78 (m, 1H), 6.90-6.91
(m, 1H), 7.01-7.05 (m, 3H), 7.27-7.30 (m, 1H), 7.43-7.45 (m, 1H),
7.88-7.98 (m, 3H), 8.08-8.10 (m, 1H), 8.89-8.96 (m, 2H), 12.67 (br
s, 1H); MS (ESI) m/z 524 (M.sup.++1); Anal. Calcd for
C.sub.27H.sub.26ClN.sub.3O.sub.6.1/4H.sub.2O: C, 61.36; H, 5.05; N,
7.95; Cl, 6.71. Found: C, 61.49; H, 5.11; N, 7.72; Cl, 7.08.
Example 179
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(3R)-pyrrolidiny-
loxy]benzoic acid
[1092] ##STR1809##
[1093] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (460 mg,
1.21 mmol), methyl 4-[(3R)-pyrrolidinyloxy]benzoate (269 mg, 1.21
mmol), EDC.HCl (280 mg, 1.46 mmol), HOBt (197 mg, 1.46 mmol), and
Et.sub.3N (205 ml, 1.47 mmol) in THF (10 ml) was stirred at room
temperature overnight. The reaction mixture was diluted with
H.sub.2O, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (100:1 to 60:1, v/v) as eluent to give methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]-(3R)-pyrrolidin-
yloxy]benzoate (555 mg, 78%) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.05-2.33 (m, 2H), 3.60-4.07 (series of s and
m, total 12H), 4.96-5.01 (m, 1H), 6.76-6.93 (m, 5H), 7.28-7.30 (m,
1H), 7.48-7.58 (m, 3H), 7.92-7.99 (m, 3H), 8.12-8.16 (m, 1H); MS
(ESI) m/z 582 (M.sup.+).
[1094] To a stirred solution of methyl
4-[1-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetyl]X.sup.3R)-pyrrol-
idinyloxy]benzoate (555 mg, 0.95 mmol) in THF (5 ml) was added 0.5
N NaOH (5 ml) and the reaction mixture was heated under reflux for
5 hr. After cooled to room temperature, the mixture was poured into
ice-1 N HCl and the resulting precipitate was collected under a
reduced pressure. The crude solid was recrystallized from
MeOH--CHCl.sub.3-Et.sub.2O to give 187 (330 mg, 61%) as a white
crystalline powder. MW 568.42 mp 175-177.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.99-2.27 (m, 2H), 3.56-3.63 (m, 4H),
3.71-3.76 (m, 1H), 3.83-3.89 (series of s and m, 4H), 5.12 and 5.20
(each m, total 1H), 6.74-6.78 (m, 1H), 6.89-7.05 (m, 4H), 7.30-7.35
(m, 1H), 7.59-7.61 (m, 1H), 7.88-7.98 (m, 4H), 8.74-8.76 (m, 1H),
8.92-8.95 (m, 1H), 12.68 (br s, 1H); MS (ESI) m/z 569 (M.sup.++1);
Anal. Calcd for C.sub.27H.sub.26BrN.sub.3O.sub.6: C, 57.05; H,
4.61; N, 7.39; Br, 14.06. Found: C, 56.91; H, 4.66; N, 7.20; Br,
14.59.
Example 180
[1095] ##STR1810##
[1096] 1 gram of Tentagel PHB resin (loading 0.29 mmol/gm) was
taken up in DMF 25 mL and Fmoc-(4-carboxymethyl)-piperidine (318
mg, 0.87 mmol) was added. The resin was shaken for 5 min then DIC
(220 mg, 0.27 mL, 1.74 mmol) and DMAP (106 mg, 0.87 mmol) were
added and the resin was shaken for 24 hr.
[1097] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and Fmoc-L-morpholine-2-carboxylic acid (307 mg,
0.87 mmol) was added. The resin was shaken for 5 min then PyBroP
(406 mg, 0.87 mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) were
added and the resin was shaken for 24 hr.
[1098] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and 4-tolylureidophenylacetic acid (247 mg, 0.87
mmol) was added and the resin was shaken for 5 min. PyBroP (406 mg,
0.87 mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) was added and the
resin was shaken for 24 hr.
[1099] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2 and shaken
for 4 hr. The resin was drained and the eluate collected. The resin
was taken up in fresh CH.sub.2Cl.sub.2 and shaken for 30 min. The
resin was drained and the eluate collected and combined with the
first fraction. The solvent was removed under vacuum and the
residue was recrystallized from ethyl acetate-hexane, yielding 85
mg 188.
Example 181
[1100] ##STR1811##
[1101] 1 gram of Tentagel PHB resin (loading 0.29 mmol/gm) was
taken up in DMF 25 mL and Fmoc-(4-carboxymethyl)-piperidine (318
mg, 0.87 mmol) was added. The resin was shaken for 5 min then DIC
(220 mg, 0.27 mL, 1.74 mmol) and DMAP (106 mg, 0.87 mmol) were
added and the resin was shaken for 24 hr.
[1102] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test The resin was taken
up in 25 mL of DMF and Fmoc-L-4-phenylproline (307 mg, 0.87 mmol)
was added. The resin was shaken for 5 min then PyBroP (406 mg, 0.87
mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) were added and the
resin was shaken for 24 hr.
[1103] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and 4-o-tolylureidophenylacetic acid (247 mg,
0.87 mmol) was added and the resin was shaken for 5 min. PyBroP
(406 mg, 0.87 mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) was added
and the resin shaken for 24 hr.
[1104] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2 and shaken
for 4 hr. The resin was drained and the eluate collected. The resin
was taken up in fresh CH.sub.2Cl.sub.2 and shaken for 30 min. The
resin was drained and the eluate collected and combined with the
first fraction. The solvent was removed under vacuum and the
residue was recrystallized from ethyl acetate-hexane, yielding 82
mg 189.
Example 182
[1105] ##STR1812##
[1106] 1 gram of Tentagel PHB resin (loading 0.29 mmol/gm) was
taken up in DMF 25 mL and Fmoc-4-carboxymethyl-piperazine (318 mg,
0.87 mmol) was added. The resin was shaken for 5 min DIC (220 mg,
0.27 mL, 1.74 mmol) and DMAP (106 mg, 0.87 mmol) were added and the
resin was shaken for 24 hr.
[1107] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and Fmoc-L-proline (294 mg, 0.87 mmol) was
added. The resin was shaken for 5 min. then PyBroP (406 mg, 0.87
mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) were added and the
resin was shaken for 24 hr.
[1108] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test The resin was taken
up in 25 mL of DMF and 4+tolylureidophenyl acetic acid (247 mg,
0.87 mmol) was added and the resin was shaken for 5 min. PyBroP
(406 mg, 0.87 mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) was added
and the resin was shaken for 24 hr.
[1109] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2 and shaken
for 4 hr. The resin was drained and the eluate collected. The resin
was taken up in fresh CH.sub.2Cl.sub.2 and shaken for 30 min. The
resin was drained and the eluate collected and combined with the
first fraction. The solvent was removed under vacuum and the
residue was recrystallized from ethyl acetate-hexane, yielding 78
mg 190.
Example 183
[1110] ##STR1813##
[1111] 1 gram of Tentagel PHB resin (loading 0.29 mmol/gm) was
taken up in DMF 25 mL and Fmoc-isonipocotic acid (306 mg, 0.87
mmol) was added. The resin was shaken for 5 min then DIC (220 mg,
0.27 mL, 1.74 mmol) and DMAP (106 mg, 0.87 mmol) were added and the
resin was shaken for 24 hr.
[1112] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and Fmoc-L-proline (294 mg, 0.87 mmol) was
added. The resin was shaken for 5 min then PyBroP (406 mg, 0.87
mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) were added and the
resin was shaken for 24 hr.
[1113] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and 4-o-tolylureidophenyl acetic acid (247 mg,
0.87 mmol) was added and the resin was shaken for 5 min. PyBroP
(406 mg, 0.87 mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) was added
and the resin was shaken for 24 hr.
[1114] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2 and shaken
for 4 hr. The resin was drained and the eluate collected. The resin
was taken up in fresh CH.sub.2Cl.sub.2 and shaken for 30 min. The
resin was drained and the eluate collected and combined with the
first fraction. The solvent was removed under vacuum and the
residue was recrystallized from ethyl acetate-hexane, yielding 73
mg 191.
Example 184
[1115] ##STR1814##
[1116] 1 gram of Tentagel PHB resin (loading 0.29 mmol/gm) was
taken up in DMF 25 mL and Fmoc-4-carboxymethyl)-piperidine (318 mg,
0.87 mmol) was added. The resin was shaken for 5 min then DIC (220
mg, 0.27 mL, 1.74 mmol) and DMAP (106 mg, 0.87 mmol) were added and
the resin was shaken for 24 hr.
[1117] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and Fmoc-L-proline (294 mg, 0.87 mmol) was
added. The resin was shaken for 5 min. then PyBroP (406 mg, 0.87
mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) were added and the
resin was shaken for 24 hr.
[1118] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in 25 mL of DMF and Fmoc-3-amino-2-oxo-1-pyrrolidineacetate (331
mg, 0.87 mmol) was added and the resin was shaken for 5 min. PyBroP
(406 mg, 0.87 mmol) and DIEA (123 mg, 0.15 mL, 0.87 mmol) was added
and the resin was shaken for 24 hr.
[1119] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was taken up in 20% piperidine in DMF and shaken for 4 hr.
The resin was drained and washed with DMF (3.times.), CH.sub.3OH
(3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried under vacuum.
The resin gave a positive bromophenylblue test. The resin was taken
up in CH.sub.2Cl.sub.2 and o-tolyl isocyanate (193 mg, 1.45 mmol,
0.18 mL) was added. The resin was shaken for 24 hr.
[1120] The resin was drained and washed with DMF (3.times.),
CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.) then dried
under vacuum. The resin gave a negative bromophenylblue test. The
resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2 and shaken
for 4 hr. The resin was drained and the eluate collected. The resin
was taken up in fresh CH.sub.2Cl.sub.2 and shaken for 30 min. The
resin was drained and the eluate collected and combined with the
first fraction. The solvent was removed under vacuum and the
residue was recrystallized from ethyl acetate-hexane, yielding 69
mg 192.
Example 185
[1121] ##STR1815##
[1122] In a 250 mL round-bottomed flask was placed
o-tolylisothiocyanate (10.0 g, 67.1 mmol) in 150 mL of
CH.sub.2Cl.sub.2. This solution was cooled to minus 78.degree. C.
and ammonia gas (excess) was bubbled through for 10 min. A
precipitate immediately formed and was found to be the desired
product o-tolylthiourea. The reaction mixture was filtered and the
solid collected by washing thoroughly with cooled CH.sub.2Cl.sub.2.
The white solid was dried under vacuum to provide 10.12 g (92%
yield) of the desired o-tolylthiourea.
[1123] The o-tolylthiourea (10.12 g, 61 mmol) was then methylated
by addition of methyl iodide (9.1 g, 62 mmol) in anhydrous methanol
(100 mL). The reaction was stirred at room temp for 6 hr and then
concentrated in vacuo. The residue was poured into aqueous ammonium
chloride and extracted 3.times. with EtOAc. The combined organics
were dried and concentrated in vacuo to give 8.7 g (84% yield) of
2-methyl-2-thio-o-tolylpseudourea. The pseudourea (8.7 g, 51 mmol)
was dissolved in methanol (100 mL) and piperidine (8.7 g, 102 mmol)
at room temp. The mixture was stirred overnight and then
concentrated in vacuo to afford 9.2 g of the product ester as a
pale yellow solid. This solid was saponified with LiOH to give 9.0
g of the desired final carboxylic acid 193.
Example 186
[1124] ##STR1816##
[1125] Methyl-4-aminophenylacetate (4.0 g, 25 mmol) was dissolved
in CH.sub.2Cl.sub.2 (100 mL) and to this solution was added
o-tolylisothiocyante (3.7 g, 25 mmol). The reaction mixture was
heated to reflux for 4 hr and then cooled to room temp. The
solution was poured in to 1N HCl and then extracted 3.times. with
EtOAc, dried over MgSO.sub.4, and concentrated in vacuo to afford
5.2 g (67% yield) of thiourea methyl ester. The ester was
saponified using LiOH to give 5.0 g of the desired
4-(o-tolylthioureido)phenylacetic acid 194.
Example 187
[1126] ##STR1817##
[1127] A solution of ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S,3R,4R)-3,4-
-isopropylidenedioxy-2-pyrrolidinylcarbonyl]-1-piperazinylacetate
(1.27 g, 1.99 mmol) in sat. HCl (gash MeOH (20 mL) was stirred at
room temp. for 2 hr, and MeOH was evaporated off. The residue was
taken up with sat. NaHCO.sub.3 solution, and extracted with
CHCl.sub.3-MeOH (4:1, v/v). The extracts were washed with brine,
dried over MgSO.sub.4, and concentrated to dryness. Chromatography
of the residue with CHCl.sub.3-MeOH (5:1, v/v) as eluent gave ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S,3R,4R)-3,4-
-dihydroxy-2-pyrrolidinyl carbonyl]-1-piperazinylacetate (990 mg,
83%) as a yellow amorphous solid. IR (KBr) 3338, 2937, 2830, 1743,
1625, 1600, 1532, 1454 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) d 1.25
(t, J=7.1 Hz, 3H), 2.20 (s, 3H), 2.46-2.56 (m, 4H), 3.15 (s, 2H),
3.40-3.72 (m, 9H), 3.62 (s, 3H), 4.01-4.08 (m, 2H), 4.16 (q, J=7.1
Hz, 2H), 4.22 (m, 1H), 4.72 (d, J=2.9 Hz, 1H), 6.68 (d, J=7.6 Hz,
1H), 6.72 (s, 1H), 7.06 (t, J=7.6 Hz, 1H), 7.15-7.18 (m, 3H),
7.52-7.56 (m, 2H), 7.90 (d, J=8.3 Hz, 1H); MS (FAB) m/z 598
(M.sup.++1).
[1128] To a solution of ethyl
4-[1-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2S,3R,4R)-3,4-
-dihydroxy-2-pyrrolidinylcarbonyl]-1-piperazinylacetate (870 mg,
1.46 mmol) in THF (15 mL) was added 0.25N NaOH (7.00 mL, 1.75
mmol). After being stirred at room temp. for 3.5 hr, the reaction
mixture was concentrated. The residue was diluted with water and
neutralized with 1 N HCl at 0.degree. C. The mixture was
concentrated and purified by ion-exchanged resin (HP-20, Mitsubishi
Chemical) to give 195 (645 mg, 78%) as a colorless amorphous solid.
IR (KBr) 3330, 2937, 1627, 1535, 1454 cm.sup.-1; .sup.1H NMR
(DMSO-d.sub.6) d 2.25 (s, 3H), 2.38 (m, 1H), 2.42-2.58 (m, 2H),
2.64 (m, 1H), 3.01 (s, 2H), 3.13-3.71 (m, 8H), 3.88 (s, 3H), 3.89
(m, 1H), 4.05 (m, 1H), 4.58 (d, J=3.2 Hz, 1H), 6.76 (dd, J=8.3, 1.5
Hz, 1H), 6.91-6.95 (m, 2H), 7.10-7.16 (m, 2H), 7.79 (d, J=8.3 Hz,
1H), 8.00 (d, J=8.3 Hz, 1H), 8.49 (s, 1H), 8.57 (s, 1H); MS (FAB)
m/z 570 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.35N.sub.5O.sub.8.2.75H.sub.2O: C, 54.32; H, 6.59; N,
11.31. Found: C, 54.07; H, 6.11; N, 11.00.
Example 188
[1129] ##STR1818##
[1130] To a suspension of 2-amino-4-thiazoleacetic acid (4 g, 25
mmol) in 1:1 CH.sub.2Cl:acetone (100 mL) was added
o-tolylisocyanate (3.5 g, 26 mmol). The mixture was heated to
reflux for 8 hr at which time a yellow precipitate had formed. the
precipitate was filtered and the solid washed generously with 1:1
CH.sub.2Cl.sub.2:acetone. The solid was recrystallized with hot
methanol and dried under vacuum to yield 4.8 g (66% yield) of the
desired 2-(o-tolylureido)-4-thiazoleacetic acid 196.
Example 189
[1131] ##STR1819##
[1132] In a round bottom flask, 3-bromobenzyl amine (3.00 g, 16.13
mmole) was dissolved in dioxane-water (1:1) and solid
Na.sub.2CO.sub.3 was added till the pH was 8-9. Boc.sub.2O (3.87 g,
17.74 mmole) was added and the reaction was stirred for 12 hr at
room temp. The reaction mixture was poured into 1N HCl and the
aqueous layer was extracted 3.times. with ethyl acetate. The
combined organic layers were then washed with water, brine then
dried over anhydrous MgSO.sub.4. The solution was filtered and the
solvent was removed under reduced pressure. The product was
purified by flash chromatography. (4:1 hexane-ethyl acetate) Yield
4.39 g 197. ##STR1820##
[1133] The Boc-protected benzyl amine (2.00 g, 6.99 mmole) was
dissolved in dry THF under argon. The reaction was cooled to minus
78.degree. C. Lithium bis(trimethylsilyl)amide (13.98 mL, 13.98
mmole) was added over 10 min. The reaction was stirred for one hr
at minus 78.degree. C. then iodomethane (1.98 g, 13.98 mmole, 0.87
mL) was added rapidly. The reaction was allowed to slowly warm to
room temp and stir overnight. The reaction was poured into 1N HCl
and the aqueous layer was extracted 3.times. with ethyl acetate.
The solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (7:1
hexane-ethyl acetate) Yield 1.68 g 198. ##STR1821##
[1134] In a pressure tube was placed the Boc-protected
3-bromobenzyl methylamine (1.68 g, 5.60 mmole). The tube was then
charged with DMF, sodium acetate (0.51 g 6.16 mmole),
P(o-tolyl).sub.3 (0.51 g, 6.16 mmole), and Pd(OAc).sub.2 (0.25 g,
1.12 mmole) The tube was flushed with argon for 10 min then methyl
acrylate (0.53 g, 0.53 mmole, 0.55 mL) was added. The tube was
sealed and heated to 135.degree. C. for 24 hr. The reaction was
cooled to 0.degree. C. and the tube was slowly opened. The solution
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were washed with
water, brine then dried over MgSO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The product was
isolated by flash chromatography. (6:1 hexane-ethyl acetate) Yield
1.60 g 199. ##STR1822##
[1135] In a pressure tube was placed the Boc-protected
3-bromobenzyl methylamine (10.00 g, 3.33 mmole). The tube was then
charged with DMF, sodium acetate (0.30 g, 3.36 mmole),
P(o-tolyl).sub.3 (0.20, 0.66 mmole), and Pd(OAc).sub.2 (0.15 g,
0.66 mmole). The tube was flushed with argon for 10 min then methyl
methacrylate (0.37 g, 3.66 mmole, 0.39 mL) was added. The tube was
sealed and heated to 135.degree. C. for 24 hr. The reaction was
cooled to 0.degree. C. and the tube was slowly opened. The solution
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were washed with
water, brine then dried over MgSO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The product was
isolated by flash chromatography. (6:1 hexane-ethyl acetate) Yield
1.01 g 200. ##STR1823##
[1136] The .alpha.,.beta.-unsaturated ester (1.60 g, 5.49 mmole)
was placed in a Paar vessel and dissolved in ethyl acetate. Pd/C
(0.3 g) was added and the vessel was pressured to 50 psi with
H.sub.2. The vessel was agitated for 12 hr. The Paar vessel was
flushed with argon and the catalyst was removed by filtration
through celite. The solvent was removed under reduced pressure.
Yield 1.60 g 201. ##STR1824##
[1137] The .alpha.-methyl-,.beta.-unsaturated ester (1.01 g, 3.16
mmole) was placed in a Paar vessel and dissolved in ethyl acetate.
Pd/C was added and the vessel was pressured to 50 psi with H.sub.2.
The vessel was agitated for 12 hr. The Paar vessel was flushed with
argon and the catalyst was removed by filtration through celite.
The solvent was removed under reduced pressure. Yield 996.41 mg
202. ##STR1825##
[1138] The Boc ester (304 mg, 1.04 mmole) was taken up in
CH.sub.2Cl.sub.2 and excess trifluoroacetic acid was added. The
reaction was then stirred for 2 hr. The solvent was removed and the
residue was taken up in ethyl acetate and washed with sat.
NaHCO.sub.3 solution. The organic layer was washed with water,
brine then dried over Na.sub.2SO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The residue was
taken up in CH.sub.2C.sub.2-DMF and HOBt (154.30 mg, 1.14 mmole),
4-[N'-(o-tolylurea)phenylacetic acid (324.11 mg, 1.14 mmole) and
EDCI (218.53 mg, 1.14 mmole) were added. The reaction was stirred
for 24 hr. The solution was poured into 1N HCl and the aqueous
layer was extracted 3.times. with ethyl acetate. The combined
organic layers were washed with water, brine then dried over
MgSO.sub.4. The solution was filtered and the solvent was removed
under reduced pressure. The product was isolated by flash
chromatography. (ethyl acetate) Yield 380.32 mg 203.
##STR1826##
[1139] The ester (380.32 mg, 0.80 mmole) was taken up in
ethanol-water (4:1) and NaOH was added. The reaction was then
heated to 50.degree. C. for 2 hr. The TLC (ethyl acetate) showed no
starting material present. The reaction was cooled to room temp.
The solution was poured into 1N HCl and the aqueous layer was
extracted 3.times. ethyl acetate. The combined organic layers were
washed with water, brine then dried over MgSO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The residue was recrystallized from ethyl acetate-hexane. Yield
319.40 mg 204.
Example 190
[1140] ##STR1827##
[1141] The Boc ester (209.60, 0.65 mmole) was taken up in
CH.sub.2Cl.sub.2 and excess trifluoroacetic acid was added. The
reaction was then stirred for 2 hr. The solvent was removed and the
residue was taken up in ethyl acetate and washed with sat.
NaHCO.sub.3 solution. The organic layer was washed with water,
brine then dried over Na.sub.2SO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The residue was
taken up in CH.sub.2Cl.sub.2-DMF and HOBt (97.45 mg, 0.72 mmole),
4-[N'-o-tolylurea)phenylacetic acid (204.70 mg, 0.72 mmole) and
EDCI (138.03 mg, 0.72 mmole) were added. The reaction was stirred
for 24 hr. The solution was poured into 1N HCl and the aqueous
layer was extracted 3.times. with ethyl acetate. The combined
organic layers were washed with water, brine then dried over
MgSO.sub.4. The solution was filtered and the solvent was removed
under reduced pressure. The product was isolated by flash
chromatography. (ethyl acetate) Yield 237.8 mg 205. ##STR1828##
[1142] The ester (237.8 mg, 0.49 mmole) was taken up in
ethanol-water (4:1) and NaOH added. The reaction was heated to
50.degree. C. for 2 hr. The TLC (ethyl acetate) showed no starting
material present. The reaction was cooled to room temp. The
solution was poured into 1N HCl and the aqueous layer was extracted
3.times. with ethyl acetate. The combined organic layers were
washed with water, brine then dried over MgSO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The residue was recrystallized from ethyl acetate-hexane. Yield
207.8 mg 206.
Example 191
[1143] ##STR1829##
[1144] The 1,2,3,4-tetrahydroisoquinoline (12.20 g, 91.60 mmole)
was taken up in H.sub.2SO.sub.4 (40 mL) and cooled to minus
10.degree. C. Concentrated HNO.sub.3 (9.0 mL) was slowly added to
the solution while maintaining the internal temp at minus
10.degree. C. On completion of the addition the reaction was
allowed to stand and slowly warm to room temp over 12 hr. The
reaction mixture was slowly added to ice and the aqueous solution
was basified with NH.sub.4OH, The aqueous layer was extracted 4
times with CHCl.sub.3. The combined organic layers were washed with
water then dried over Na.sub.2SO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The resulting
brown oil was taken up in ethanol and concentrated HCl was added.
The resulting white solid was collected by filtration and dried
under vacuum. Yield 8.0 g 207. ##STR1830##
[1145] The 6-nitro-1,2,3,4-tetrahydroisoquinoline (1.00 g, 5.61
mmole) was taken up in ethanol. Methyl bromoacetate (0.86 g, 5.61
mmole, 0.53 mL) and triethylamine (1.17 g, 11.59 mmole, 1.62 mL)
were then added and the mixture was heated to reflux for 5 hr. The
solution was cooled to room temp and the solution was concentrated
under vacuum. The solution was added to water and the aqueous layer
was extracted 3.times. with ethyl acetate. The combined organic
layers were dried over Na.sub.2SO.sub.4, filtered and the solvent
was removed under reduced pressure. The product was isolated by
flash chromatography (3:1 hexane-ethyl acetate). Yield 702 mg 208.
##STR1831##
[1146] The above ester (702 mg, 2.8 mmole) was placed in a Paar
vessel and dissolved in ethanol. Pd/C (100 mg) was added and the
vessel was pressured to 50 psi with H.sub.2. The vessel was
agitated for 24 hr. The Paar vessel was flushed with argon and the
catalyst was removed by filtration through celite. The solvent was
removed under reduced pressure. .sup.1H-NMR showed only desired
product. Yield 587 mg 209. ##STR1832##
[1147] The aniline (587.0 mg, 2.66 mmole) was taken up in dry
CH.sub.2Cl.sub.2 and pyridine under argon. The reaction was cooled
to 0.degree. C. A CH.sub.2Cl.sub.2 solution of
3-methoxy-4-(N'phenylureido)phenylacetyl chloride (837.70 mg, 2.66
mmole) was added over 5 min. The reaction was then allowed to warm
to room temp and stir overnight. The reaction mixture was then
poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were then washed
with sat. NaHCO.sub.3, water, brine then dried over MgSO.sub.4. The
solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography (ethyl
acetate). Yield 618.36 mg 210. ##STR1833##
[1148] The methyl ester (618.36 mg, 1.20 mmole) above was taken up
in THF--H.sub.2O and LiOH (558.07 mg, 13.30 mmole) was added. The
reaction mixture was stirred at room temp for 24 hr. The reaction
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were then washed
with water, brine then dried over MgSO.sub.4. The solution was
filtered and the solvent was removed under reduced pressure. The
product was purified by recrystallization. (hexane-ethyl acetate).
Yield 600 mg 211.
Example 192
[1149] ##STR1834##
[1150] The 3-nitro-phenyl propionic acid (1.00 g, 5.12 mmole) was
taken up in dry THF under argon. The reaction was cooled to
0.degree. C. and BH.sub.3-THF (1.0M, 15.37 mmole, 15.37 mL) was
added over 10 min. The reaction was stirred at 0.degree. C. for 1
hr then slowly quenched with water. The solution was slowly warmed
to room temp then poured into 1N HCl. The aqueous layer was
extracted 3.times. with ethyl acetate. The combined organic layers
were then washed with sat. NaHCO.sub.3, water, brine then dried
over MgSO.sub.4. The solution was filtered and the solvent was
removed under reduced pressure. The product was isolated by flash
chromatography (1:1 hexane-ethyl acetate) Yield 909.0 mg 212.
##STR1835##
[1151] The 3-nitro-phenyl propanol (909.0 mg, 5.02 mmole) was taken
up in dry CH.sub.2Cl.sub.2. In a second round bottom flask
(COCl).sub.2 (700.65 mg, 5.52 mmole, 0.48 mL) was added to dry
CH.sub.2Cl.sub.2 under argon. The (COCl).sub.2--CH.sub.2Cl.sub.2
solution was then cooled to minus 60.degree. C. and DMSO (862.56
mg, 11.4 mmole, 0.78 mL) was slowly added. The reaction was stirred
at minus 60.degree. C. for 5 min then the alcohol solution was
added via a cannula over 5 min. The reaction mixture was stirred at
minus 60.degree. C. for 1 hr then Et.sub.3N (2.54 g, 25.10 mmole,
3.50 mL) was added and the reaction was allowed to slowly warm to
room temp. The reaction was poured into 1N HCl and the aqueous
layer was extracted 3.times. with ethyl acetate. The combined
organic layers were washed with sat. NaHCO.sub.3, water, brine then
dried over MgSO.sub.4. The solution was filtered and the solvent
was removed under reduced pressure. H1-NMR showed no starting
material present. The aldehyde 213 was used as is without further
purification. ##STR1836##
[1152] In a round bottom flask NaH (132.48 mg, 5.52 mmole) was
slurried in dry THF under argon. Triethyl 2-phosphonopropionate
(1.31 g, 5.52 mmole, 1.18 mL) dissolve in dry THF was added slowly
via a syringe. The reaction mixture was stirred for 30 min at room
temp. The above aldehyde, dissolved in dry THF under argon, was
added to the phosphonate solution via syringe over 10 min. The
reaction mixture was stirred for 12 hr. The reaction was poured
into 1N HCl and the aqueous layer was extracted 3.times. with ethyl
acetate. The combined organic layers were washed with sat.
NaHCO.sub.3, water, brine then dried over MgSO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The product was isolated by flash chromatography. (1:1 ethyl
acetate-hexane) Yield 992.0 mg 214. ##STR1837##
[1153] The above .alpha.,.beta.-unsaturated ester (992.0 mg, 3.77
mmole) was placed in a Paar vessel and dissolved in ethanol. The
vessel was flushed with argon and Pd/C (200.0 mg) was added. The
argon atmosphere was replaced with H.sub.2 at 50 psi. The Paar
vessel was then shaken for 12 hr. The hydrogen was flushed from the
vessel with argon and the catalyst was removed by filtration
through celite. The solvent was removed under reduced pressure.
.sup.1H-NMR showed only the desired product. Yield 851.2 mg 215.
##STR1838##
[1154] The above aniline (850.0 mg, 3.61 mmole) was taken up in dry
CH.sub.2Cl.sub.2 and pyridine under argon. The reaction was cooled
to 0.degree. C. A CH.sub.2Cl.sub.2 solution of 3-methoxy-4
(N'phenylureido)phenylacetyl chloride (1.14 g, 3.61 mmole) was
added over 5 min. The reaction was then allowed to warm to room
temp and stir overnight The reaction mixture was then poured into
1N HCl and the aqueous layer was extracted 3.times. with ethyl
acetate. The combined organic layers were then washed with sat.
NaHCO.sub.3, water, brine then dried over MgSO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The product was isolated by flash chromatography (ethyl acetate)
Yield 576.0 mg 216. ##STR1839##
[1155] The above ethyl ester (576.0 mg, mmole) was taken up in
ethanol-water and NaOH was added. The reaction mixture was heated
to 50.degree. C. for 2 hr. The reaction was cooled to room temp and
then poured into 1N HCl. The aqueous layer was extracted 3.times.
times with ethyl acetate. The combined organic layers were then
washed with water, brine then dried over MgSO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The product was purified by Sep-Pak column. Yield 534 mg 217.
Example 193
[1156] ##STR1840##
[1157] One gram of Wang resin (tentagel S--PHB 0.3 mmole loading)
was suspended in a solution of 3-Fmoc-amino)-phenylpropionic acid
(361.31 mg, 0.90 mmole), DMAP (109.95 mg, 0.90 mmole), HOBt 243.63
mg, 0.90 mmole), and DIC (227.16 mg, 1.80 mmole, 0.28 mL) in a
mixture of DMF and CH.sub.2Cl.sub.2. The mixture was shaken for 20
hr and drained. The resin 218 was washed with DMF, MeOH,
CH.sub.2Cl.sub.2 and dried under reduced pressure. ##STR1841##
[1158] To the above resin (500 mg, 0.15 mmole) was added a solution
of piperidine-DMF (50% v/v, 4 mL) and the mixture was shaken for 4
hr. The resin was washed with DMF, MeOH, CH.sub.2Cl.sub.2. To the
resin was added TMOF and isobutrylaldehyde (108.17 mg, 1.50 mmole,
0.14 mL). The mixture was shaken for 4 hr. The resin was drained
and fresh TMOF and isobutrylaldehyde was added. The mixture was
then shaken for 12 hr. The resin was drained and taken up in
MeOH-1% AcOH and NaCNBH.sub.3 (150.0 mg, 2.39 mmole) was added The
resin was shaken for 6 hr. The resin was drained and washed with
MeOH, MeOH-Et.sub.3N, MeOH, DMF, CH.sub.2Cl.sub.2. The resin was
taken up in DMF and 3-methoxy-4-(N'-phenylureido)phenylacetic acid
(141.45 mg, 0.45 mmole), PyBrop (209.78 mg, 0.45 mmole), and DIEA
(58.16 mg, 0.45 mmole, 0.08 mL) were added. The resin was then
shaken for 24 hr then drained. The resin was washed with DMF, MeOH,
CH.sub.2Cl.sub.2. To the resin was added a solution of TFA in
CH.sub.2Cl.sub.2 (30% v/v 3 mL) and the mixture was shaken for 5
hr. The mixture was filtered and the filtrate was concentrated in
vacuo. The residue was purified by Sep-Pak column. After removal of
the solvent, Et.sub.2O was added to the residue and the solid was
collected to afford 15 mg 219 as a crystalline solid
Example 194
[1159] ##STR1842##
[1160] Tentagel PHB resin (1.0 g, loading 0.29 mmole/gm) was taken
up in 25 mL of DMF and 6-bromohexanoic acid (169 mg, 0.87 mmol) was
added. The resin was shaken for 5 min then DIC (220 mg, 0.27 mL,
1.74 mmoles) and DMAP (35 mg, 0.29 mmole) were added and the resin
was shaken for 14 hr. The resin was drained and washed with DMF
(3.times.), CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.)
then dried under vacuum.
[1161] To this resin was added
2,2-dimethyl-1,3-dioxolane-4-methanamine (227 mg, 1.74 mmol) and
lithium iodide (232 mg, 1.74 mmol) in 15 mL of DMF. The resin was
shaken for 14 hr at room temp. The resin was drained and washed
with DMF (3.times.), CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2
(3.times.) then dried under vacuum. The resin gave a positive
bromophenylblue test.
[1162] The resin was taken up in 25 mL of DMF and
4-o-tolylureido-3-methoxyphenylacetic acid (247 mg, 0.87 mmole) was
added and the resin was shaken for 5 min. PyBrOP (406 mg, 0.87
mmole) and DIEA (123 mg, 0.15 mL, 0.87 mmole) was added and the
resin was shaken for 14 hr. The resin was drained and washed with
DMF (3.times.), CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2
(3.times.) then dried under vacuum. The resin gave a negative
bromophenylblue test.
[1163] The resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2
and shaken for 4 hr. The resin was drained and the elutant
collected. The resin was taken up in fresh CH.sub.2Cl.sub.2 and
shaken for 30 min. The resin was drained and the elutant collected
and combined with the first fraction. The solvent was removed under
vacuum and the residue was recrystallized from ethyl
acetate-hexane, yield 56 mg 220.
Example 195
[1164] ##STR1843##
[1165] 1 gram of Tentagel PHB resin (loading 0.29 mmole/gm) was
taken up in DMF 25 mL and Fmoc-7-aminoheptanoic acid (319 mg, 0.87
mmole) was added. The resin was shaken for 5 min then DIC (220 mg,
0.27 mL, 1.74 mmol) and DMAP (106 mg, 0.87 mmole) were added and
the resin was shaken for 24 hr. The resin was drained and washed
with DMF (3.times.), CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2
(3.times.) then dried under vacuum. The resin gave a negative
bromophenylblue test. The resin was taken up in 20% piperidine in
DMF and shaken for 4 hr. The resin was drained and washed with DMF
(3.times.), CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2 (3.times.)
then dried under vacuum. The resin gave a positive bromophenylblue
test.
[1166] The resin was taken up in 25 mL of DMF and
4-tolylureido-3-methoxyphenylacetic acid (247 mg, 0.87 mmole) was
added and the resin was shaken for 5 min. PyBrOP (406 mg, 0.87
mmole) and DIEA (123 mg, 0.15 mL, 0.87 mmole) was added and the
resin was shaken for 14 hr. The resin was drained and washed with
DMF (3.times.), CH.sub.3OH (3.times.) and CH.sub.2Cl.sub.2
(3.times.) then dried under vacuum. The resin gave a negative
bromophenylblue test.
[1167] The resin was then taken up in 90% TFA in CH.sub.2Cl.sub.2
and shaken for 4 hr. The resin was drained and the elutant
collected. The resin was taken up in fresh CH.sub.2Cl.sub.2 and
shaken for 30 min. The resin was drained and the elutant collected
and combined with the first fraction. The solvent was removed under
vacuum and the residue was recrystallized from ethyl
acetate-hexane, yield 66 mg 221.
Example 196
[1168] ##STR1844##
[1169] To a solution of oxalyl chloride (3.8 g, 30 mmol) in
CH.sub.2Cl.sub.2 (100 mL) was added DMSO (2.4 g, 31 mmol) dropwise
over 30 min at minus 78.degree. C. To this solution was added
N-Boc-prolinol (5.0 g, 25 mmol) dropwise over 15 min. The reaction
was stirred at minus 78.degree. C. for 3 hr and then quenched by
the cold addition of 1N HCl, extracted 3.times. with EtOAc, dried,
and concentrated in vacuo to afford the crude prolinal which was
chromatographed (25% EtOAc/hexanes) to yield 3.8 g of the desired
product.
[1170] A solution of Methyl (triphenylphosphoranylidene)butanoate
(6.9 g, 19 mmol) in THF (100 mL) was generated. LiHMDS (10 mL of a
2.0M soln, 20 mmol) was added at minus 78.degree. C. and then
stirred for 1 hr. The above prolinal (3.8 g, 19 mmol) was then
added in one portion and the mixture was allowed to warm to room
temp over 4 hr. The reaction was quenched by the addition of 1N
HCl, extracted 3.times. with EtOAc, dried, and concentrated in
vacuo to afford the crude alkene which was chromatographed (25%
EtOAc/hexanes) to yield 2.9 g of the desired product.
[1171] Hydrogenation of the alkene was performed by placing the
alkene (2.9 g, 10 mmol) in ethanol (20 mL) and adding a catalytic
amount of 10% Pd/C followed by Parr hydrogenation at 40 psi for 4
hr, the resulting alkane was used without purification. The Boc
group was removed by the addition of 1:1 TFA/CH.sub.2Cl.sub.2 at
room temp. The reaction was stirred for 2 hr and the solvent was
removed in vacuo. The crude amine 1.9 g was used without further
purification.
[1172] A solution of the above free amine (1.9 g, 10 mmol) in
CH.sub.2Cl.sub.2 (100 mL) was generated. To this solution was added
EDCI (2.95 g, 10 mmol), DMAP (1.2 g, 10 mmol), and
4-tolylureido-3-methoxyphenylacetic acid (3.15 g, 10 mmol) at room
temp. The reaction mixture was stirred for 4 hr and then quenched
by the addition of 1 N HCl, extracted 3.times. with EtOAc, dried,
and concentrated in vacuo. The crude amide was chromatographed (5%
MeOH/CH.sub.2Cl.sub.2) to yield 1.95 g of the desired product.
[1173] The ester (1.95 g, 4.2 mmol) was taken up in 1:1
THF--H.sub.2O and LiOH was added at room temp. The reaction mixture
was then stirred for 3 hr. The solution was poured into 1N HCl and
the aqueous layer was extracted 3.times. with EtOAc. The combine
organic layers were washed with water, brine and dried over
anhydrous magnesium sulfate. The solution was filtered and the
solvent removed under reduced pressure. The solid was then
triturated with cold ether to give 1.65 g of the desired carboxylic
acid 222.
Example 197
[1174] ##STR1845##
[1175] To a solution of methyl 8-aminooctanoate (2.0 g, 12 mmol) in
1:1 dioxane:water (100 mL) was added Boc anhydride (2.8 g, 13 mmol)
and K.sub.2CO.sub.3 (10 g). This solution was allowed to stir at
room temp for 14 hr. The reaction was then poured onto 1 N HCl,
extracted 3.times. with EtOAc, dried, and concentrated in vacuo.
The crude carbamate was chromatographed (50% EtOAc/hexanes) to
yield 2.7 g of the desired product.
[1176] The Boc-protected amine was methylated by placing it in THF
(75 mL), followed by the addition of LiHMDS (25 mL of a 2.0M soln.,
50 mmol) at minus 78.degree. C., this solution was then stirred for
30 min and methyl iodide (7.2 g, 50 mmol) was added in one portion
the reaction mixture was allowed to warm to room temp overnight.
The reaction was quenched by the addition of 1 N HCl, extracted
3.times. with EtOAc, dried, and concentrated in vacuo. The crude
methylated carbamate was chromatographed (50% EtOAc/hexanes) to
yield 1.9 g of the desired dimethyl product.
[1177] The Boc group was removed by the addition of 1:1
TFA/CH.sub.2Cl.sub.2 at room temp. The reaction was stirred for 2
hr and the solvent was removed in vacuo. The crude amine 900 mg was
used without further purification.
[1178] A solution of the above free amine (900 mg, 4.5 mmol) in
CH.sub.2Cl.sub.2 (100 mL) was generated. To this solution was added
EDCI (1.33 g, 4.5 mmol), DMAP (567 mg, 4.5 mmol), and
4+tolylureido-3-methoxyphenylacetic acid (1.45 g, 4.6 mmol) at room
temp. The reaction mixture was stirred for 4 hr and then quenched
by the addition of 1 N HCl, extracted 3.times. with EtOAc, dried,
and concentrated in vacuo. The crude amide was chromatographed (5%
MeOH/CH.sub.2Cl.sub.2) to yield 1.2 g of the desired product.
[1179] The ester (1.2 g, 2.4 mmol) was taken up in 1:1
THF--H.sub.2O and LiOH was added at room temp. The reaction mixture
was then stirred for 3 hr. The solution was poured into 1N HCl and
the aqueous layer was extracted 3.times. with EtOAc. The combine
organic layers were washed with water, brine and dried over
anhydrous magnesium sulfate. The solution was filtered and the
solvent removed under reduced pressure. The solid was then
triturated with cold ether to give 1.01 g of the desired carboxylic
acid 223.
Example 198
[1180] ##STR1846##
[1181] To a suspension of 4-aminophenylacetic acid (10 g, 66 mmol)
in 1:1 CH.sub.2Cl.sub.2:acetone (100 mL) was added o-tolyisocyanate
(8.8 g, 66 mmol). The mixture was heated to reflux for 4 hr at
which time a white precipitate had formed. The precipitate was
filtered and the solid washed generously with 1:1
CH.sub.2Cl.sub.2:acetone. The solid was recrystallized with hot
methanol and dried under vacuum to yield 14.1 g (75% yield) of the
desired 4-o-tolylureido)phenylacetic acid 224.
Example 199
[1182] ##STR1847##
[1183] To a suspension of 2-amino-4-thiazoleacetic acid (4 g, 25
mmol) in 1:1 CH.sub.2Cl:acetone (100 mL) was added
o-tolylisocyanate (3.5 g, 26 mmol). The mixture was heated to
reflux for 8 hr at which time a yellow precipitate had formed. the
precipitate was filtered and the solid washed generously with 1:1
CH.sub.2Cl.sub.2:acetone. The solid was recrystallized with hot
methanol and dried under vacuum to yield 4.8 g (66% yield) of the
desired 2-(o-tolylureido)-4-thiazoleacetic acid 225.
Example 200
[1184] ##STR1848##
[1185] 3-Bromo-4-hydroxybenzonitrile (5.00 g, 25.25 mmol) was taken
up in DMF. Benzyl bromide (4.75 g, 27.78 mmol, 3.30 mL) and
Cs.sub.2CO.sub.3 (16.45 g, 50.50 mmol) were added and the reaction
was heated to 50.degree. C. for 2 hr. The solution was cooled to
room temperature and poured into 1NHCl. The aqueous layer was
extracted 3.times. with ethyl acetate. The combined organic layers
were washed with water, brine then dried over MgSO.sub.4. The
solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (hexane
to 8:1 hexane-ethyl acetate) Yield 8.90 g 226. ##STR1849##
[1186] 3-Bromo-4-benyloxybenzonitrile (1.50 g, 5.21 mmol) was taken
up in dry THF under argon and the solution was cooled to 0.degree.
C. BH.sub.3-THF (10.4 mL, 10.41 mmol) was added via syringe over 5
min. The reaction mixture was then warmed to room temp then heated
to reflux for 12 hr. The solution was cooled to 0.degree. C. and
methanol was slowly added. When no more gas evolution was observed
the solution was warmed to room temp and excess 1N NaOH solution
was added. Boc.sub.2O (1.25 g, 5.73 mmol) was added and the
reaction mixture was stirred at room temp for 12 hr. The solution
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were washed with
water, brine then dried over MgSO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The product was
isolated by flash chromatography. (7:1 ethyl acetate-hexane) Yield
1.80 g 227 ##STR1850##
[1187] The Boc-protected benzyl amine (1.80 g, 4.59 mmol) was
dissolved in dry THF under argon. The reaction was cooled to minus
78.degree. C. Lithium bis(trimethylsilyl)amide (13.77 mL, 13.77
mmol) was added over 10 min. The reaction was stirred for 1 hr at
minus 78.degree. C., then iodomethane (1.95 mL, 13.77 mmol, 0.86
mL) was added rapidly. The reaction was allowed to slowly warm to
room temp and stir overnight. The reaction was poured into 1N HCl
and the aqueous layer was extracted 3.times. with ethyl acetate.
The solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (7:1
hexane-ethyl acetate) Yield 1.70 g 228. ##STR1851##
[1188] In a pressure tube was placed the
4-(N-methyl-Boc-aminomethyl)-2-bromobenzyloxy phenol (1.70 g, 4.18
mmol). The tube was then charged with DMF, sodium acetate (0.38 g,
4.60 mmol), dppp (0.35 g, 0.84 mmol), and Pd(OAc).sub.2 (0.19 g,
0.84 mmol) The tube was flushed with argon for 10 min and then
methyl acrylate (0.40 g, 4.60 mmol, 0.41 mL) was added. The tube
was sealed and heated to 135.degree. C. for 24 hr. The reaction was
cooled to 0.degree. C. and the tube was slowly opened. The solution
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were washed with
water, brine then dried over MgSO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The product was
isolated by flash chromatography. (6:1 hexane-ethyl acetate) Yield
1.12 g 229 ##STR1852##
[1189] The unsaturated ester (307.40 mg, 0.75 mmol) was taken up in
CH.sub.2Cl.sub.2 and excess TFA was added. The reaction was stirred
for 4 hr at room temp. The solvent was removed under reduced
pressure and the residue was dried under high vacuum. The solvent
was removed and the residue was taken up in ethyl acetate and
washed with sat. NaHCO.sub.3 solution. The organic layer was washed
with water, brine then dried over Na.sub.2SO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The residue was taken up in CH.sub.2Cl.sub.2-DMF and HOBt (110.99
mg, 0.82 mmol), 3-methoxy-4-(N'-phenylureido)phenylacetic acid
(258.31 mg, 0.82 mmol) and EDCI (157.20 mg, 0.82 mmol) were added.
The reaction was stirred for 24 hr. The solution was poured into 1N
HCl and the aqueous layer was extracted 3.times. with ethyl
acetate. The combined organic layers were washed with water, brine
then dried over MgSO.sub.4. The solution was filtered and the
solvent was removed under reduced pressure. The product was
isolated by flash chromatography. (ethyl acetate) Yield 296.30 mg
230 ##STR1853##
[1190] The unsaturated ester (296.30 mg, 0.49 mmol) was taken up in
EtOAc and Pd/C (75 mg) was added under argon. The argon atmosphere
was replaced with hydrogen at 1 atmosphere and stirred for 24 hr.
The hydrogen atmosphere was removed and replaced with argon. The
catalyst was removed by filtration through celite and the celite
pad was washed with ethyl acetate 3.times.. The solvent was removed
under reduced pressure. H.sup.1-NMR showed only the desired
product. No further purification was needed. Yield 233.00 mg 231
##STR1854##
[1191] The ester (233.00 mg, 0.45 mmol) was taken up in
THF--H.sub.2O (4:1) and LiOH (94.41 mg, 2.25 mmol) was added. The
reaction mixture was stirred at room temp for 24 hr. The solution
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were washed with
water, brine then dried over MgSO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The product was
washed with ether-hexane (1:1) and dried under high vacuum. Yield
211.58 mg 232
Example 201
[1192] ##STR1855##
[1193] The carboxylic acid (65.00 mg, 0.13 mmol) was taken up in
benzene and para-toluenesulfonic acid (10.00 mg, 0.06 mmol) was
added. A Dean-Stark trap was added and the solution was heated to
reflux for 24 hr. The reaction was cooled to room temp and poured
into sat. NaHCO.sub.3. The organic layer was separated and the
aqueous layer was extracted 3.times. with ethyl acetate. The
combined organic layers were washed with water, brine and dried
over MgSO.sub.4. The solution was filtered and the solvent was
removed under reduced pressure. The product was isolated by flash
chromatography (4:1 hexane-ethyl acetate to ethyl acetate) Yield
29.00 mg 233
Example 202
[1194] ##STR1856##
[1195] 5-Bromonicotinic acid (5.15 g, 25.49 mmol) was taken up in
EtOH and H.sub.2SO.sub.4 (1 mL) was added and the solution heated
to reflux for 24 hr. The solution was cooled to rt and
concentrated. The solution was then added to sat. NaHCO.sub.3 and
the aqueous layer were extracted 3.times. with Et.sub.2O. The
combined organic layers were dried over Na.sub.2SO.sub.4, filtered
and the solvent was removed under reduced pressure. The product was
sufficiently pure for the next step. Yield 5.42 g 234
##STR1857##
[1196] The ethyl 5-Bromonicotinate (5.40 g, 23.47 mmol) was taken
up in 95% EtOH and NaBH.sub.4 (8.31 g, 225.69 mmol) was added
slowly at room temp. After addition the solution was stirred for 24
hr at room temp. Water was slowly added to the solution, then the
mixture was stirred for 4 hr. The EtOH was removed under reduced
pressure and the aqueous layer was extracted 3.times. with
CH.sub.2Cl.sub.2. The combined organic layers were dried over
Na.sub.2SO.sub.4, filtered and the solvent was removed under
reduced pressure. The product was isolated by flash chromatography.
(2:1 ethyl acetate-hexane) Yield 2.12 g 235 ##STR1858##
[1197] The benzyl alcohol (2.12 g, 11.28 mmol) was taken up
Et.sub.2O and HCl.sub.(g) was bubbled through the solution for 10
min. The solution was stirred at room temp for 1 hr and then the
solid was collected by filtration. The solid was washed with
Et.sub.2O and the then dried. The HCl salt was added to SOCl.sub.2
and the mixture was heated to reflux for 1.5 hr. The solution was
cooled to room temp and Et.sub.2O was added to precipitate the
product. The solid was collected by filtration, washed with
Et.sub.2O and dried under vacuum. Yield 2.42 g 236 ##STR1859##
[1198] The benzyl chloride (2.42 g, 9.96 mmol) was added over 1 hr
to CH.sub.3NH.sub.2 (75.9 mL, 2.5M in EtOH) at room temp. The
reaction was stirred at room temp for 48 hr. The solution was
concentrated and added to sat. NaHCO.sub.3. The aqueous layer was
extracted 3.times. with ethyl acetate. The combined organic layers
were dried over Na.sub.2SO.sub.4, filtered and the solvent was
removed under reduced pressure. Yield 1.19 g 237 ##STR1860##
[1199] The 3-bromo-5-(N-methyl-aminomethyl)-pyridine (1.19 g, 5.01
mmol) was taken up in DMF and triethylamine (0.90 g, 1.24 mL, 8.89
mmol) was added. Boc.sub.2O (1.55 g, 7.10 mmol) was added and the
reaction mixture was stirred at room temp for 48 hr. The solution
was poured into 1N HCl and the aqueous layer was extracted 3.times.
with ethyl acetate. The combined organic layers were washed with
water, brine then dried over MgSO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The product was
isolated by flash chromatography (2% methanol-CH.sub.2Cl.sub.2)
Yield 1.6 g 238 ##STR1861##
[1200] The sodium salt of a-methyl acrylic acid (5.00 g, 46.27
mmol) was dissolved in DMF and benzyl bromide (8.70 g, 50.89 mmol)
was added at room temp. Potassium carbonate (7.03 g, 50.89 mmol)
was then added and the solution was heated to 50.degree. C. for 24
hr. The solution was poured into 1N HCl and the aqueous layer was
extracted 3.times. with diethyl ether. The combined organic layers
were washed with water, brine then dried over MgSO.sub.4. The
solution was filtered and the solvent was removed carefully under
reduced pressure. The product was isolated by flash chromatography
(2% ether-pentane) Yield 6.93 g 239 ##STR1862##
[1201] In a pressure tube was placed the
3-bromo-5-(N-methyl-Boc-aminomethyl)-pyridine (700.00 mg, 2.33
mmol). The tube was then charged with DMF, triethylamine (260.05
mg, 2.57 mmol, 0.36 mL), dppp (193.85 mg, 0.47 mmol), and
Pd(OAc).sub.2 (105.52 mg, 0.47 mmol) The tube was flushed with
argon for 10 min then benzyl methacrylate (452.86 mg, 2.57 mmol)
was added. The tube was sealed and heated to 135.degree. C. for 24
hr. The reaction was cooled to 0.degree. C. and the tube was slowly
opened. The solution was poured into 1N HCl and the aqueous layer
was extracted 3.times. with ethyl acetate. The combined organic
layers were washed with water, brine then dried over MgSO.sub.4.
The solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (6:1
hexane-ethyl acetate) Yield 785.23 mg 240 ##STR1863##
[1202] The unsaturated ester (392.61 mg, 0.99 mmol) was taken up in
CH.sub.2Cl.sub.2 and excess TFA was added. The reaction was stirred
for 4 hr at room temp. The solvent was removed under reduced
pressure and the residue was dried under high vacuum. The solvent
was removed and the residue was taken up in ethyl acetate and
washed with sat. NaHCO.sub.3 solution. The organic layer was washed
with water, brine then dried over Na.sub.2SO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The residue was taken up in CH.sub.2Cl.sub.2-DMF and HOBt (147.53
mg, 1.09 mmol), 3-methoxy-4-(N'-phenylureido)phenylacetic acid
(342.64 mg, 1.09 mmol) and EDCI (208.96 mg, 1.09 mmol) were added.
The reaction was stirred for 24 hr. The solution was poured into 1N
HCl and the aqueous layer was extracted 3.times. with ethyl
acetate. The combined organic layers were washed with water, brine
then dried over MgSO.sub.4. The solution was filtered and the
solvent was removed under reduced pressure. The product was
isolated by flash chromatography. (ethyl acetate) 363.79 mg 241
##STR1864##
[1203] The unsaturated ester (363.00 mg, 0.61 mmol) was taken up in
CH.sub.3OH and Pd/C (100.00 mg) was added under argon. The argon
atmosphere was replaced with hydrogen at 1 atmosphere and stirred
for 24 hr. The hydrogen atmosphere was removed and replaced with
argon. The catalyst was removed by filtration through celite and
the celite pad was washed with ethyl acetate 3.times.. The solvent
was removed under reduced pressure. H.sup.1-NMR showed only the
desired product. The solid was washed with ether and then dried
under high vacuum. Yield 254.79 mg 242
Example 203
[1204] ##STR1865##
[1205] 3-Cyanobenzaldehyde (9.41 g, 71.76 mmol) was taken up in
ethanol and cooled to 0.degree. C. The NaBH.sub.4 (2.71 g, 71.76
mmol) was added in small portions. The solutions was stirred for 30
min at 0.degree. C. then allowed to warm to room temp and stirred
for 1 hr. The reaction was slowly poured into 1NHCl and the aqueous
layer was extracted 3.times. with ethyl acetate. The combined
organic layers were washed with water, brine then dried over
MgSO.sub.4. The solution was filtered and the solvent was removed
under reduced pressure. The residue was taken up in DMF and
imidazole (2.08 g, 30.50 mmol) was added. TBDPSCl (4.61 g, 16.78
mmol, 4.36 mL) was then added and the reaction was stirred at room
temp for 12 hr. The solution was poured into 1N HCl and the aqueous
layer was extracted 3.times. with ethyl acetate. The combined
organic layers were washed with water, brine then dried over
MgSO.sub.4. The solution was filtered and the solvent was removed
under reduced pressure. The product was isolated by flash
chromatography. (7:1 hexane-ethyl acetate to 4:1 hexane-ethyl
acetate) Yield 16.23 g 243 ##STR1866##
[1206] The silyl protected 3-cyanobenzyl alcohol (8.50 g, 34.36
mmol) was taken up in ethyl acetate and Boc-O (8.25 g, 37.79 mmol)
was added. Pd/C (1.0 g) was added and the Parr vessel was
pressurized with hydrogen at 50 psi. The vessel was shaken for 24
hr then the hydrogen was flushed with argon and the catalyst was
removed by filtration through a celite pad. The celite was washed
3.times. with ethyl acetate. The solvent was removed under reduced
pressure and the product was isolated by flash chromatography (10:1
hexane-ethyl acetate) Yield 11.10 g 244 ##STR1867##
[1207] The O-silyl-N-Boc-protected benzyl alcohol (5.00 g, 14.22
mmol) was dissolved in dry THF under argon. The reaction was cooled
to minus 78.degree. C. Lithium bis(trimethylsilyl)amide (42.67 mL,
42.67 mmol) was added over 10 min. The reaction was stirred for 1
hr at minus 78.degree. C. then iodomethane (6.06 g, 42.67 mmol,
2.66 mL) was added rapidly. The reaction was allowed to slowly warm
to room temp and stir overnight. The reaction was poured into 1N
HCl and the aqueous layer was extracted 3.times. with ethyl
acetate. The solution was filtered and the solvent was removed
under reduced pressure. The product was isolated by flash
chromatography. (2% ethyl acetate-hexane) Yield 4.7 g 245
##STR1868##
[1208] The O-silyl-Boc-N-methyl protected benzyl alcohol (4.7 g,
9.60 mmol) was taken up in THF and TBAF (14.39 mL, 1.0M in THF) at
room temp. The solution was stirred for 4 hr. TLC showed no
starting material present The reaction was poured into 1N HCl and
the aqueous layer was extracted 3.times. with ethyl acetate. The
solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (4:1
hexane-ethyl acetate to 1:1 hexane-ethyl acetate) Yield 2.39 g 246
##STR1869##
[1209] The N-methyl Boc protected benzyl alcohol (1.00 g, 3.98
mmol) was taken up in dry CH.sub.2Cl.sub.2 under argon.
Triphenylphosphine (1.46 g, 5.57 mmol) was added and the solution
was cooled to 0.degree. C. Carbon tetrabromide (1.85 g, 5.57 mmol)
dissolved in dry CH.sub.2Cl.sub.2 was added over 10 min. The
solution was stirred for 1 h at 0.degree. C. then the solvent was
removed under reduced pressure. The residue was taken up in
Et.sub.2O and the resulting solid was removed by filtration and the
filtrate was collected and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography (2%
ether-pentane) Yield 1.15 g 247 ##STR1870##
[1210] LHMDS (3.23 mL, 3.23 mmol) was added to dry DME under argon
at minus 78.degree. C. Methyl butyrate (300 mg, 2.94 mmol, 0.33 mL)
dissolved in dry DME was added to the LHMDS over 15 min and the
solution was stirred for 1 hr at minus 78.degree. C.
3-N-methyl-N-Boc protected benzyl bromide (1.02 g, 3.23 mmol)
dissolved in dry DME was added to the enolate solution over 15 min
then the solution was allowed to slowly warm to minus 20.degree. C.
and stirred for 4 hr. The reaction was poured into 1N HCl and the
aqueous layer was extracted 3.times. with ethyl acetate. The
solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (3%
ethyl acetate-hexane) Yield 414 mg 248. ##STR1871##
[1211] The Boc ester (121.60 mg, 0.36 mmol) was taken up in
CH.sub.2Cl.sub.2 and excess trifluoroacetic acid was added. The
reaction was then stirred for 2 hr. The solvent was removed and the
residue was taken up in ethyl acetate and washed with sat.
NaHCO.sub.3 solution. The organic layer was washed with water,
brine then dried over Na.sub.2SO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The residue was
taken up in CH.sub.2Cl.sub.2-DMF and HOBt (54.10 mg, 0.40 mmol)
3-methoxy-4-(N'-phenylureido)phenylacetic acid (125.74 mg, 0.40
mmol) and EDCI (77.0 mg, 0.40 mmol) were added. The reaction was
stirred for 24 hr. The solution was poured into 1N HCl and the
aqueous layer was extracted 3.times. with ethyl acetate. The
combined organic layers were washed with water, brine then dried
over MgSO.sub.4. The solution was filtered and the solvent was
removed under reduced pressure. The product was isolated by flash
chromatography. (ethyl acetate) Yield 165.20 mg 249 ##STR1872##
[1212] The ester (165.20, 0.31 mmol) was taken up in ethanol-water
(4:1) and NaOH was added. The reaction was then heated to
50.degree. C. for 2 hr. The TLC (ethyl acetate) showed no starting
material present. The reaction was cooled to room temp. The
solution was poured into 1N HCl and the aqueous layer was extracted
3.times. with ethyl acetate. The combined organic layers were
washed with water, brine then dried over MgSO.sub.4. The solution
was filtered and the solvent was removed under reduced pressure.
The residue was recrystallized from ethyl acetate-hexane. Yield
120.00 mg 250
Example 204
[1213] ##STR1873##
[1214] Butyrolactone (250 mg, 2.90 mmol, 223.20 mL) was added to
LHMDS (2.90 mL, 1.0M in hexane) in THF at minus 78.degree. C. under
argon over 10 min. The solution was stirred at minus 78.degree. C.
for 1 hr. 3-N-methyl-N-Boc protected benzyl bromide (991.24 mg,
2.90 mmol) dissolved in dry DME was added to the enolate solution
over 15 min then the solution was allowed to slowly warm to room
temp and stirred for 12 hr. The reaction was poured into 1N HCl and
the aqueous layer was extracted 3.times. with ethyl acetate. The
solution was filtered and the solvent was removed under reduced
pressure. The product was isolated by flash chromatography. (4:1
hexane-ethyl acetate to 1:1 ethyl acetate-hexane) Yield 501.18 mg
251 ##STR1874##
[1215] The Boc ester (250.00 mg, 0.78 mmol) was taken up in
CH.sub.2Cl.sub.2 and excess trifluoroacetic acid was added. The
reaction was then stirred for 2 hr. The solvent was removed and the
residue was taken up in ethyl acetate and washed with sat.
NaHCO.sub.3 solution. The organic layer was washed with water,
brine then dried over Na.sub.2SO.sub.4. The solution was filtered
and the solvent was removed under reduced pressure. The residue was
taken up in CH.sub.2Cl.sub.2-DMF and HOBt (116.40 mg, 0.86 mmol)
3-methoxy-4-(N'-phenylureido)phenylacetic acid (270.33 mg, 0.86
mmol) and EDCI (165.06 mg, 0.86 mmol) were added. The reaction was
stirred for 24 hr. The solution was poured into 1N HCl and the
aqueous layer was extracted 3.times. with ethyl acetate. The
combined organic layers were washed with water, brine then dried
over MgSO.sub.4. The solution was filtered and the solvent was
removed under reduced pressure. The product was isolated by flash
chromatography. (ethyl acetate) Yield 119.00 mg 252
Example 205
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-met-
hylaminoethoxy]benzoic acid
[1216] ##STR1875##
[1217] To a stirred and cooled (0.degree. C.) solution of N-methyl
ethanolamine (3.10 g, 41.27 mmol), Et.sub.3N (11.80 mL, 84.66 mmol)
in DMF-H2O (3:1, v/v, 40 mL) was added dropwise 30% toluene
solution of benzyl chloroformate (25.40 g, 49.13 mmol) for over 15
min. The resulting mixture was stirred for 1 day at room temp. The
mixture was extracted with EtOAc. The extract was washed with sat.
NaHCO.sub.3, brine, dried over Na.sub.2SO.sub.4, and evaporated.
The residue was purified by column chromatography on silica-gel
with n-hexane:EtOAc (3:1, v/v) then CHCl.sub.3 as eluent to give
4.67 g (54%) N-methyl-N-(benzyloxy carbonyl)ethanolamine as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) d 1.82 (bs, 1H), 3.00 (s,
3H), 3.46 (bs, 2H), 3.77 (bs, 2H), 5.13 (s, 2H), 7.29-7.36 (m,
5H).
[1218] To a stirred solution of ethyl 4-hydroxy-3-methoxybenzoate
(2.01 g, 10.25 mmol), N-methyl-N-(benzyloxycarbonyl)ethanolamine
(2.11 g, 10.08 mmol), PPh.sub.3 (3.26 g, 12.43 mmol) in THF was
added DIAD (2.65 mL, 13.46 mmol) and the reaction mixture was
heated under reflux overnight. The mixture was evaporated, and the
residue was subjected to short column chromatography on silica-gel
with n-hexane/EtOAc (5:1, v/v) as eluent to give ethyl
3-methoxy-4-[2-methyl-2-(benzyloxycarbonyl)aminoethoxy]benzoate as
a crude product.
[1219] To a solution of the crude product (5.20 g, 13.42 mmol) in
EtOH (50 mL) was added AcOH (5 mL) and the solution was
hydrogenated over 5% Pd/C for 4 hr. The mixture was filtered to
remove the catalyst and the filtrate was evaporated. The residue
was diluted with CHCl.sub.3 and washed with sat. NaHCO.sub.3,
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with CHCl.sub.3:MeOH (10:1, v/v) as
eluent to give 510 mg (2 steps 20%) ethyl
3-methoxy-4-(2-methylamino ethoxy)benzoate as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) d 1.39 (t, 3H, J=7.3 Hz), 1.82 (bs, 1H),
2.52 (s, 3H), 3.04 (t, 2H, J=5.3 Hz), 3.91 (s, 3H), 4.18 (t, 2H,
J=5.3 Hz), 4.36 (q, 2H, J=7.3 Hz), 6.90 (d, 1H, J=8.3 Hz), 7.55 (d,
1H, J=2.0 Hz), 7.65 (dd, 1H, J=2.0, 8.3 Hz).
[1220] To a stirred solution of ethyl
3-methoxy-4-(2-methylaminoethoxy)benzoate (510 mg, 2.01 mmol) in
DMF (13 mL) was added pentafluorophenyl ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (900 mg,
1.87 mmol) and Et.sub.3N (0.420 mL, 3.01 mmol), and the resulting
mixture was stirred for 2 days. The mixture was diluted with EtOAc,
washed with 1 N HCl, sat. NaHCO.sub.3, brine, and dried over
Na.sub.2SO.sub.4. After being evaporated, the residue was purified
by column chromatography on silica-gel with CHCl.sub.3:MeOH (50:1,
v/v) to give 880 mg (85%) ethyl
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-me-
thylaminoethoxy]benzoate as a colorless amorphous solid.
.sup.1H-NMR (CDCl.sub.3) d 1.37-1.41 (m, 3H), 2.28 (s, 3H), 3.03
and 3.18 (s, 3H), 3.56 (s, 2H), 3.65 (s, 2H), 3.75-3.87 (m, 6H),
4.06-4.24 (2H, m), 4.33-4.39 (m, 2H), 6.68-8.08 (series of m,
12H).
[1221] To a solution of ethyl
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-me-
thylaminoethoxy]benzoate (880 mg, 1.601 mmol) in THF (15 mL) was
added 0.25 N NaOH (15 mL). Then the reaction mixture was heated
under reflux overnight. The mixture was poured into 1 N HCl (100
mL), and the solid was collected. The crude solid was
recrystallized from MeOH--CHCl.sub.3 to give 253 as a white powder.
IR (KBr) 1700 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) d 2.25 (s, 3H),
2.50 (s, 2H), 2.91 and 3.12 (s, 3H) 3.53-3.76 (m, 2H), 3.80 (s,
3H), 3.84 (s, 3H), 4.16-4.21 (m, 2H), 6.72-8.56 (series of m, 12H),
12.68 (bs, 1H); MS (FAB) m/z 522 (M.sup.++1); Anal. Calcd. for
C.sub.28H.sub.31N.sub.3O.sub.7.1H.sub.2O: C, 62.33; H, 6.16; N,
6.63. Found: C, 62.17; H, 6.05; N, 7.57.
Example 206
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]eth-
oxy]isophthalic acid
[1222] ##STR1876##
[1223] To a stirred solution of
N-methyl-N-benzyloxycarbonylethanolamine (1.05 g, 5.02 mmol),
dimethyl 4-hydroxy isophthalate (1.05 g, 5.00 mmol), Ph.sub.3P
(1.59 g, 6.06 mmol) in THF (20 mL) was added DIAD (1.28 mL, 6.50
mmol) at room temp. The resulting mixture was then heated under
reflux overnight. After cooling to room temp, the mixture was
evaporated. The residue was dissolved in EtOH and added 5% Pd/C
(200 mg). The stirred resulting mixture was hydrogenated for 2 hr
at 1 atm. The mixture was filtered to remove the catalyst, and the
filtrate was evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (30:1, v/v) as
eluent to give 480 mg (36% for 2 steps) dimethyl
4-(2-methylaminoethoxy)isophthalate as an oil. .sup.1H-NMR
(CDCl.sub.3) d 1.68 (s, 1H), 2.53 (s, 3H), 3.01-3.04 (m, 2H), 3.89
(s, 3H), 3.90 (s, 3H), 4.21-4.23 (m, 2H), 7.00 (d, 1H, J=8.8 Hz),
8.14 (dd, 1H J=2.4, 8.8 Hz), 8.50 (d, 1H, J=2.4 Hz); MS (FAB), m/z
268 (M.sup.++1).
[1224] To a stirred solution of dimethyl
4-(2-methylaminoethoxy)isophthalate (410 mg, 1.53 mmol) in DMF (13
mL) was added pentafluorophenyl ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (700 mg,
1.46 mmol) and Et.sub.3N (340 .mu.l, 2.44 mmol), and the resulting
mixture was stirred overnight. The mixture was diluted with EtOAc,
washed with 1 N HCl, sat. NaHCO.sub.3, and brine. The solution was
dried over Na.sub.2SO.sub.4 and evaporated to give 780 mg (95%)
dimethyl
4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino
ethoxy]isophthalate as a crystalline powder. .sup.1H-NMR
(CDCl.sub.3) d 2.29 (s, 3H), 3.24 (s, 3H), 3.59 (s, 3H), 3.67-3.68
(m, 2H), 3.84 (s, 3H), 3.91 (s, 3H), 3.81-3.86 (m, 2H), 4.25-4.28
(m, 2H), 6.51-8.48 (series of m, 12H); MS (FAB) m/z 564
(M.sup.++1).
[1225] To a solution of dimethyl
4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylaminoetho-
xy]isophthalate (780 mg, 1.384 mmol) in THF (30 mL) was added 0.25
N NaOH (30 mL). The resulting mixture was then heated under reflux
overnight. The mixture was poured into ice-1 N HCl (200 mL) and the
solid was collected. The crude solid was recrystallized from
MeOH--CHCl.sub.3 to give 420 mg (57%)
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]methylamino]ethoxy] isophthalic acid 254 as a white
crystalline powder. mp 139-141.degree. C. IR (KBr) 1700 cm.sup.-1;
.sup.1H-NMR (DMSO-d.sub.6) d 2.94 (s, 3H), 3.18 (s, 3H) 3.62-3.86
(m, total 8H), 4.24-4.28 (m, 2H), 6.74-8.58 (series of m, total
12H), 12.91 (bs, 1H); MS (FAB) m/z 536 (M.sup.++1); Anal. Calcd.
for C.sub.28H.sub.29N.sub.3O.sub.8.2.5HCl: C, 53.66; H, 5.07; N,
6.70. Found: C, 53.80; H, 4.64; N, 6.70.
Example 207
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]aminoe-
thoxy]benzoic acid
[1226] ##STR1877##
[1227] To a solution of 2-ethanolamine (5.16 g, 84.48 mmol),
Et.sub.3N (23.50 mL, 168.60 mL) in dioxane-H.sub.2O (1/1, 160 mL)
was added dropwise (Boc).sub.2O (23.40 mL, 101.86 mmol) at room
temp. The reaction mixture was stirred for 2 days at room temp. The
resulting mixture was diluted with CHCl.sub.3, washed with 0.5 N
HCl, sat. NaHCO.sub.3, and brine. The separated organic layer was
dried over Na.sub.2SO.sub.4 and evaporated to give 11.86 g (87%)
N-Boc-2-ethanolamine as an oil. .sup.1H-NMR (CDCl.sub.3) d 1.45 (s,
9H), 3.29-3.31 (m, 2H), 3.71-3.72 (m, 2H).
[1228] To a stirred solution of ethyl 4-hydroxy-3-methoxybenzoate
(1.46 g, 7.44 mmol), N-Boc ethanolamine (1.19 g, 7.38 mmol),
PPh.sub.3 (2.53 g, 9.65 mmol) in THF (30 mL) was added DIAD (1.90
mL, 9.65 mmol), and the resulting mixture was then heated under
reflux overnight. The mixture was evaporated to give a crude gum.
The crude product was dissolved in CH.sub.2Cl.sub.2 (20 mL) and TFA
(20 mL). The resulting mixture was stirred for 2.5 hr at room temp.
The mixture was concentrated in vacuo and the residue was made
basic with sat. NaHCO.sub.3 and extracted with CHCl.sub.3. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
evaporated to give the 1.61 g (90% for 2 steps) ethyl
3-methoxy-4-(2-aminoethoxy)benzoate as a yellow oil. .sup.1H-NMR
(CDCl.sub.3) d 1.39 (t, 3H, J=7.3 Hz), 3.14-3.17 (m, 2H), 3.92 (s,
3H), 4.09-4.11 (m, 2H), 4.36 (q, 2H, J=7.3 Hz), 6.89 (d, 1H, J=8.3
Hz), 7.56 (d, 1H, J=2.0 Hz), 7.66 (dd, 1H, J=2.0, 8.3 Hz).
[1229] To a stirred solution of ethyl
3-methoxy-4-(2-aminoethoxy)benzoate (250 mg, 1.04 mmol) and
pentafluorophenyl ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (500 mg,
1.04 mmol) was added Et.sub.3N (210 .mu.l, 3.01 mmol), and the
resulting mixture was stirred for 2 days. 0.25 N NaOH (20 mL) and
THF (20 mL) was added to the mixture and the resulting mixture was
heated under reflux overnight. After cooling, the mixture was
evaporated and the residue was acidified by the addition of 1 N
HCl. The mixture was extracted with CHCl.sub.3, and the extract was
washed with brine, dried over Na.sub.2SO.sub.4, and evaporated. The
obtained crude solid was recrystallized from CHCl.sub.3 to give 110
mg (20% for 2 steps)
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]amino-
ethoxy]benzoic acid 255 as a white crystalline powder. mp
180-181.degree. C.; IR (KBr) 1687 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) d 2.24 (s, 3H), 3.37 (s, 2H), 3.38 (s, 2H),
3.41-3.50 (m, 2H), 3.81 (s, 3H), 3.83 (s, 3H), 4.06-4.08 (m, 2H),
6.76-8.55 (series of m, total 12H); MS (FAB) m/z 508 (M.sup.++1);
Anal. Calcd for C.sub.27H.sub.29N.sub.3O.sub.7.1/2H.sub.2O: C,
62.78; H, 5.85; N, 8.13. Found: C, 62.46; H, 5.69; N, 8.03.
Example 208
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]ethyla-
minoethoxy]benzoic acid
[1230] ##STR1878##
[1231] To a cooled (0.degree. C.) solution of ethyl
3-methoxy-4-(2-aminoethoxy)benzoate (1.93 g, 8.07 mmol) and
Et.sub.3N (2.00 mL, 14.35 mmol) was added TFAA (1.35 mL, 9.56 mmol)
and the resulting mixture was stirred overnight at room temp. The
resulting mixture was diluted with Et.sub.2O and washed
successively with sat. NaHCO.sub.3, 1 N HCl, H.sub.2O, and brine.
The extract was dried over Na.sub.2SO.sub.4 and evaporated to give
1.22 g (45%) ethyl
3-methoxy-4-(2-N-trifluoroacetamidoethoxy)benzoate as an oil.
.sup.1H-NMR (CDCl.sub.3) d 1.39 (t, 3H, J=7.3 Hz), 3.77-3.81 (m,
2H), 3.92 (s, 3H), 4.18-4.20 (m, 2H), 4.37 (q, 2H, J=7.3 Hz), 6.92
(d, 1H, J=8.7 Hz), 7.59 (d, 1H, J=2.0 Hz), 7.67 (dd, 1H, J=2.0, 8.7
Hz); MS (FAB) m/z 335 (M.sup.+), 290 (M.sup.+-OEt).
[1232] To a stirred solution of ethyl
3-methoxy-4-(2-N-trifluoroacetamidoethoxy)benzoate (1.20 g, 3.58
mmol) in DMF (15 mL) was added K.sub.2CO.sub.3 (0.98 g, 7.09 mmol)
and EtI (0.43 mL, 5.38 mmol) at room temp. The resulting mixture
was stiffed for 2 days at 60.degree. C. The mixture was diluted
with EtOAc, washed successively with 1 N HCl, brine, and dried over
Na.sub.2SO.sub.4. The solvent was evaporated and the residue was
purified by column chromatography on silica-gel with n-hexane-EtOAc
(2:1, v/v) as eluent to give 990 mg (76%) ethyl
3-methoxy-4-[2-(N-ethyl-N-trifluoroacetamido)ethoxybenzoate as a
yellow crystalline solid. .sup.1H-NMR (CDCl.sub.3) d 1.28-1.31 (m,
3H), 1.37-1.40 (m, 3H), 3.64-3.69 (m, 2H), 3.81-3.84 (m, 2H), 3.92
(s, 3H), 4.27-4.30 (m, 2H), 4.34-4.39 (m, 2H), 6.89 (d, 1H, J=8.3
Hz), 7.55 (d, 1H, J=2.0 Hz), 7.66 (dd, 1H, J=2.0, 8.3 Hz); MS (FAB)
m/z 364 (M.sup.++1).
[1233] To a stirred solution of ethyl
3-methoxy-4-[2-(N-ethyl-N-trifluoroacetamido)ethoxybenzoate (990
mg, 2.73 mmol) in THF-MeOH--H.sub.2O (2:1:1, v/v, 20 mL) was added
K.sub.2CO.sub.3 (560 mg, 4.05 mmol), and the resulting mixture was
stirred overnight. The resulting mixture was diluted with H.sub.2O
and extracted with EtOAc. The extract was washed successively with
sat. NaHCO.sub.3, brine, dried over Na.sub.2SO.sub.4, and
evaporated to give 800 mg (q.y.) ethyl
3-methoxy-4-(2-ethylaminoethoxy)benzoate as an oil. .sup.1H-NMR
(CDCl.sub.3) d 1.15 (t, 3H, J=7.3 Hz), 1.39 (t, 3H, J=7.3 Hz), 1.76
(bs, 1H), 2.74 (q, 2H, J=7.3 Hz), 3.08 (t, 2H, J=5.4 Hz), 3.91 (s,
3H), 4.18 (t, 2H, J=5.4 Hz), 4.36 (q, 2H, J=7.3 Hz), 6.90 (d, 1H,
J=8.3 Hz), 7.55 (d, 1H, J=2.0 Hz), 7.66 (dd, 1H, J=2.0, 8.3 Hz); MS
(FAB) m/z 268 (M.sup.++1).
[1234] To a stirred solution of ethyl
3-methoxy-4-(2-ethylaminoethoxy)benzoate (290 mg, 1.08 mmol) and
pentafluorophenyl ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (502 mg,
1.05 mmol) in DMF (7 mL) was added Et.sub.3N (250 ul, 1.79 mmol),
and the resulting mixture was stirred overnight. The mixture was
diluted with EtOAc, washed with 0.5 N HCl, brine, and dried over
Na.sub.2SO.sub.4. The solvent was evaporated and the residue was
purified by column chromatography on silica-gel with
CHCl.sub.3-MeOH (40:1, v/v) as an eluent to give 550 mg (93%) ethyl
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]ethyl-
aminoethoxy]benzoate as an amorphous solid. .sup.1H-NMR
(CDCl.sub.3) d 1.11-1.18 (m, 3H), 1.37-1.41 (m, 3H), 2.30 (s, 3H),
3.47-3.53 (m, 2H), 3.61-3.75 (m, 7H), 3.84 (s, 3H), 4.03-4.27 (m,
2H), 4.33-4.39 (m, 2H), 6.34-8.07 (series of m, total 12H).
[1235] To a solution of ethyl
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]ethylaminoethoxy]benzoate (550 mg, 0.98 mmol) in THF (15 mL)
was added 0.25 N NaOH (15 mL). The resulting mixture was then
heated under reflux for 2 days. The mixture was poured into 1 N HCl
and the solid was collected. The crude solid was recrystallized
from EtOH--CHCl.sub.3 to give 182 mg (35%)
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]ethyl-
aminoethoxy]benzoic acid 256 as a white crystalline powder. mp
115-118.degree. C.; IR (KBr) 1707 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) d 1.02-1.12 (m, 3H), 2.25 (s, 3H), 2.50 (s, 2H),
3.35-3.89 (m, 10H), 4.11-4.16 (m, 2H), 6.71-8.56 (series of m,
total 12H), 12.65 (br s, 1H); MS (FAB) m/z 536 (M.sup.++1); Anal.
Calcd for C.sub.29H.sub.33N.sub.3O.sub.7.3/4H.sub.2O: C, 63.43; H,
6.33; N, 7.65. Found: 63.34; H, 6.28; N, 7.28.
Example 209
3-nitro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]aminoeth-
oxy]benzoic acid
[1236] ##STR1879##
[1237] To a stirred solution of 4-hydroxy-3-nitrobenzoic acid (5.18
g, 28.29 mmol) in benzene-MeOH (4:1, v/v, 140 mL) was added
TMSCHN.sub.2 (14.10 mL, 28.20 mmol, 2 M solution in hexane) at room
temp, and the resulting mixture was stirred overnight. The mixture
was evaporated and the residue was purified by column
chromatography on silica-gel with CHCl.sub.3 as eluent to give 4.18
g (75%) methyl 3-nitro-4-hydroxybenzoate as a yellow crystalline
solid. .sup.1H-NMR (CDCl.sub.3) d 3.95 (s, 3H), 7.22 (d, 1H, J=8.8
Hz), 8.24 (dd, 1, J=2.0, 8.8 Hz), 8.83 (d, 1H, J=2.0 Hz), 10.89 (s,
1H).
[1238] To a stirred solution of methyl 3-nitro-4-hydroxybenzoate
(1.98 g, 10.04 mmol), N-Boc ethanolamine (1.63 g, 10.11 mmol) and
PPh.sub.3 (3.43 g, 13.08 mmol) in THF (40 mL) was added DIAD (2.57
mL, 13.05 mmol), and the reaction mixture was then heated under
reflux overnight The resulting mixture was evaporated to give a
gum. The residual crude gum was dissolved in CH.sub.2Cl.sub.2 (30
mL) and TFA (30 mL), and the mixture was stirred for 1 hr at room
temp. The mixture was concentrated in vacuo and made basic with
sat. NaHCO.sub.3. The mixture was extracted with CHCl.sub.3, washed
with brine, and dried over Na.sub.2SO.sub.4. The solvent was
evaporated in vacuo to give the oily residue, which was purified by
column chromatography on silica-gel with CHCl.sub.3 then
CHCl.sub.3-MeOH (20:1, v/v) as eluent to give 930 mg (27% for 2
steps) methyl 3-nitro-4-(2-aminoethoxy)benzoate as gum. .sup.1H-NMR
(CDCl.sub.3) d 3.16-3.19 (m, 1H), 3.53-3.57 (m, 1H), 3.90 and 3.94
(s, 3H), 3.95-3.98 (m, 1H), 4.21-4.24 (m, 1H), 6.89-6.91 and
7.11-7.13 (m, 1H), 8.03-8.19 and 8.21 (m, 1H), 8.52 and 8.86 (m,
1H).
[1239] To a stirred solution of pentafluorophenyl ester of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (1.86 g,
3.87 mmol) and methyl 3-nitro-4-(2-aminoethoxy)benzoic acid (0.93
g, 3.87 mmol) in DMF (27 mL) was added Et.sub.3N (0.90 mL, 6.46
mmol), and the resulting mixture was stirred overnight. The mixture
was poured into 0.5 N HCl and the resulting solid was collected.
The crude solid was dissolved in THF-0.25 N NaOH (1/1, 20 mL) and
the resulting mixture was heated under reflux overnight. The
mixture was extracted with EtOAc, washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The crude solid was
recrystallized from CHCl.sub.3-EtOH to give 60 mg (3% for 2 steps)
3-nitro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-
aminoethoxy]benzoic acid 257 as a yellow crystalline solid. mp
112-115.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) d 2.24 (s, 3H),
3.37-3.66 (m, 7H), 3.84 (s, 3H), 4.27-4.30 (m, 1H), 6.74-8.56
(series of m, total 12H); MS (FAB), m/z 523 (M.sup.++1); Anal.
Calcd for H.sub.26N.sub.4O.sub.8. 3/2H.sub.2O: C, 56.83; H, 5.32;
N, 10.20. Found: C, 56.66; H, 4.90; N, 9.33.
Example 210
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]ethyla-
minoethoxy]benzoic acid
[1240] ##STR1880##
[1241] To a stirred solution of pentafluorophenyl ester of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (135 mg,
0.28 mmol) and ethyl 3-methoxy-4-(2-ethylaminoethoxy)benzoate (78
mg, 0.29 mmol) was added Et.sub.3N (0.1 mL, 0.72 mmol), and the
resulting mixture was stirred overnight. The mixture was diluted
with EtOAc, washed successively with 0.5N HCl, brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (50:1,
v/v) as eluent to give 160 mg (q.y.) ethyl
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]ethyl-
aminoethoxy]benzoate as an oil. .sup.1H-NMR (CDCl.sub.3) d
1.13-1.23 (m, 3H), 1.37-1.40 (m, 3H), 2.90-3.89 (m, 12H), 4.09-4.28
(m, 2H), 4.33-4.39 (m, 2H), 6.70-8.21 (series of m, total 12H).
[1242] To a stirred solution of ethyl
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]ethyl-
aminoethoxy]benzoate (160 mg, 0.28 mmol) in THF (5 mL) was added
0.25 N NaOH (5 mL) and the resulting mixture was the heated under
reflux overnight. The mixture was poured into 1 N HCl and the solid
was collected. The crude solid was recrystallized from
EtOH--CHCl.sub.3-n-hexane to give 70 mg (46%)
3-methoxy-4-[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]ethyl-
aminoethoxy]benzoic acid 258 as a yellow crystalline powder. mp
105-110.degree. C.; IR (KBr) 1687 cm.sup.-1; .sup.1H-NMR
(DMSO-d.sub.6) d 1.00-1.10 (m, 3H), 2.48 (s, 2H), 3.35-3.81 (m,
10H), 4.13-4.14 (m, 2H), 6.70-9.15 (series of m, 12H); MS (FAB) m/z
540 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.30FN.sub.3O.sub.7.1/2H.sub.2O: C, 61.31; H, 5.82; N,
7.47. Found: C, 61.05; H, 5.82; N, 7.47.
Example 211
[1243] ##STR1881##
4-[4-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetyl-1-piperazinylbe-
nzoic acid
[1244] A stirred mixture of tert-butyl 1-piperazinecarboxylate
(1.00 g, 5.37 mmol), ethyl 4-fluorobenzoate (903 mg, 5.37 mmol),
and K.sub.2CO.sub.3 (1.11 g, 8.06 mmol) in DMF (10 mL) was heated
at 120.degree. C. overnight. After cooling, the mixture was diluted
with EtOAc (300 mL), followed by washing with brine (2.times.200
mL), drying over MgSO.sub.4, and evaporation. The residue was
chromatographed on silica-gel with CHCl.sub.3-EtOAc (20:1 to 4:1,
v/v) as eluent to give 257 mg (14%) ethyl
4-[4-(tert-butyloxycarbonyl)-1-piperazinyl]benzoate as a pale
yellow amorphous solid. IR (KBr) 1701, 1612 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) d 1.37 (3H, t, J=7.3 Hz), 1.49 (9H, s), 3.30 (4H, t,
J=5.4 Hz), 3.58 (4H, t, J=5.4 Hz), 4.33 (2H, q, J=7.3 Hz), 6.87
(2H, d, J=8.8 Hz), 7.94 (2H, dt, J=8.8, 2.4 Hz); MS (FAB) m/z 335
(M.sup.++1); Anal. Calcd for C.sub.18H.sub.26N.sub.2O.sub.4: C,
64.54; H, 7.84; N, 8.38. Found: C, 64.39; H, 7.89; N, 8.38.
[1245] To a stirred solution of ethyl
4-[4-(tert-butyloxycarbonyl)-1-piperazinyl]benzoate (240 mg, 0.718
mmol) in CH.sub.2Cl.sub.2 (5 mL) was added TFA (5 mL), and the
resulting mixture was stirred for 3 hr. The mixture was
concentrated in vacuo and the residue was made basic by the
addition of sat. NaHCO.sub.3, followed by extraction with
CHCl.sub.3 (2.times.100 mL). The combined extracts were dried over
Na.sub.2CO.sub.3 and evaporated to give 168 mg ethyl
4-(1-piperazinyl)benzoate (100%) as a yellow oil. .sup.1H-NMR
(CDCl.sub.3) d 1.37 (3H, t, J=7.3 Hz), 3.03 (4H, t, J=4.9 Hz), 3.29
(4H, t, J=4.9 Hz), 4.33 (2H, q, J=7.3 Hz), 6.87 (2H, dt, J=8.8, 2.4
Hz), 7.91-7.94 (2H, m).
[1246] To a stirred solution of ethyl 4-(1-piperazinyl)benzoate
(170 mg, 0.730 mmol) and
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (229 mg,
0.730 mmol) in DMF (10 mL) was added EDC.HCl (210 mg, 1.10 mmol),
DMAP (catalytic amount), and HOBt (catalytic amount), and the
mixture was stirred overnight. The mixture was poured into H.sub.2O
(100 mL) and the solid was collected with suction. The residue was
recrystallized from CHCl.sub.3-n-hexane to give 290 mg (75%) ethyl
4-[4-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetyl-1-piperazinyl
benzoate as a colorless crystalline powder. mp 208-210.degree. C.;
IR (KBr) 1711, 1695 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) d 1.37 (3H,
t, J=7.3 Hz), 2.29 (3H, s), 3.14 (2H, t, J=4.9 Hz), 3.28 (2H, t,
J=4.9 Hz), 3.62 (2H, t, J=4.9 Hz), 3.71 (3H, s), 3.72 (2H, s), 3.79
(2H, t, J=4.9 Hz), 4.33 (2H, q, J=7.3 Hz), 6.38 (1H, s), 6.78-6.99
(4H, m), 7.13-7.24 (4H, m), 7.50 (1H, d, J=7.8 Hz), 7.92 (2H, d,
J=8.8 Hz), 8.12 (1H, d, J=7.8 Hz); MS (FAB) m/z 531 (M.sup.++1);
Anal. Calcd for C.sub.30H.sub.34N.sub.4O.sub.5.0.5H.sub.2O: C,
66.77; H, 6.54; N, 10.38. Found: C, 66.89; H, 6.39; N, 10.45.
[1247] To a stirred solution of ethyl
4-[4-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-acetyl-1-piperazinyl-
benzoate (290 mg, 0.547 mmol) in MeOH-THF (2:1, v/v, 15 mL) was
added 0.25N NaOH (5 mL, 1.25 mmol) and the mixture was heated under
reflux for 3 hr. The mixture was poured into ice-1N HCl (100 mL)
and the solid was collected with suction. The residue was
recrystallized from CHCl.sub.3-MeOH to give 190 mg (69%)
4-[4-[3-methoxy-4-[N'-(2-methylphenyl)
ureido]phenyl]acetyl-1-piperazinylbenzoic acid 259 as a yellow
crystalline powder. mp 240-245.degree. C.; .sup.1H-NMR (DMSO) d
2.24 (3H, s), 3.17-3.50 (8H, m), 3.72 (2H, s), 3.86 (3H, s), 6.77
(1H, d, J=8.3 Hz), 6.90 (1H, s), 6.91-6.96 (3H, m), 7.11-7.17 (2H,
m), 7.76-7.80 (3H, m), 8.03 (1H, d, J=8.3 Hz), 8.47 (1H, s), 8.58
(1H, s), 12.30 (1H, s); Anal. Calcd for
C.sub.28H.sub.30N.sub.4O.sub.5.H.sub.2O: C, 64.60; H, 6.20; N,
10.76. Found: C, 64.64; H, 5.85; N, 10.51.
Example 212
(R)-3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylami-
no]-1-propoxy]benzoic acid
[1248] ##STR1882##
[1249] To a cooled (0.degree. C.) solution of
(R)-2-amino-1-propanol (3.01 g, 0.04 mmol) and Et.sub.3N (6.70 mL,
0.05 mmol) in DMF-H.sub.2O (1:1, v/v) (40 mL) was added
(Boc).sub.2O (10.0 mL, 0.04 mmol), and the resulting mixture was
stirred at room temp for 2 days. The mixture was diluted with
EtOAc, washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4 and
evaporated to give 6.91 g (98%)
(R)-2-N-tert-butoxycarbonylamino-1-propanol as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta.1.15 (d, 3H, J=6.8 Hz), 1.45 (s,
9H), 3.48-3.53 (m, 1H), 3.62-3.66 (m, 1H), 3.76-3.77 (m, 1H); MS
(FAB) m/z 176 (M.sup.++1), 120 (M.sup.+-.sup.tBu).
[1250] To a stirred solution of ethyl 4-hydroxy-3-methoxybenzoate
(7.74 g, 0.04 mmol), (R)-2-N-tert-butoxycarbonylamino-1-propanol
(6.91 g, 0.04 mmol) and Ph.sub.3P (13.44 g, 0.05 mmol) in THF (70
mL) was added diisopropyl azodicarboxylate (DIAD) (10.0 mL, 0.05
mmol), and the resulting mixture was heated under reflux overnight.
After cooling to room temp, the solvent was evaporated. The mixture
was dissolved in CH.sub.2Cl.sub.2 (50 mL) and TFA (30 mL) and the
solution was stirred at room temp for 1 hr. After concentration in
vacuo, the residue was poured into sat. NaHCO.sub.3 and extracted
with CHCl.sub.3. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with 5% MeOH in CHCl.sub.3 as eluent
to give 7.93 g (2 steps 79%) ethyl
(R)-3-methoxy-4-(2-amino-1-propoxy)benzoate as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.90 (d, 3H, J=6.8 Hz), 1.39 (t,
3H, J=7.3 Hz), 1.72 (bs, 2H), 3.42-3.47 (m, 1H), 3.74-3.89 (m, 1H),
3.91 (s, 3H), 3.96-4.00 (m, 1H), 4.35 (q, 2H, J=7.3 Hz), 6.88 (d,
1H, J=8.3 Hz), 7.55 (d, 1H, J=2.0 Hz), 7.65 (dd, 1H, J=2.0, 8.3
Hz); MS (FAB) m/z 254 (M.sup.++1).
[1251] To a stirred solution of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (459 mg, 0.96
mmol) and ethyl (R)-3-methoxy-4-(2-aminopropoxy)benzoate (242 mg,
0.96 mmol) in DMF (5 mL) was added Et.sub.3N (200 .mu.l, 1.43
mmol), and the resulting mixture was stirred for 2 hr. The mixture
was diluted with EtOAc, washed with 0.5 N HCl, brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as
eluent to give 360 mg (69%) ethyl
(R)-3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylam-
ino]-1-propoxy]benzoate as a colorless crystalline solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.23-1.28 (m, 3H), 1.38-1.41 (t,
3H, J=7.3 Hz), 2.32 (s, 3H), 3.50-4.13 (m, total 11H), 4.36 (q, 2H,
J=7.3 Hz), 6.65-8.13 (series of m, total 12H).
[1252] To a stirred solution of ethyl
(R)-3-methoxy-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)
ureido]phenylacetylamino]-1-propoxy]benzoate (360 mg, 0.66 mmol) in
THF-MeOH (20 mL, 9:1, v/v) was added 0.25 N NaOH (10 mL), and the
resulting mixture was heated under reflux overnight. The mixture
was poured into ice-1 N HCl, and precipitate was collected. The
crude solid was recrystallized from CHCl.sub.3-n-hexane to give 172
mg (50%) 260 as a white crystalline powder. mp 168-169.degree. C.;
IR (KBr) 1687 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.18
(d, 3H, J=6.8 Hz), 2.24 (s, 3H), 2.50-2.51 (m, 2H), 3.80 (s, 3H),
3.84 (s, 3H), 3.87-4.06 (m, 2H), 4.07-4.14 (m, 1H), 6.76-8.57
(series of m, total 12H), 12.66 (bs, 1H); MS (FAB) m/z 522
(M.sup.++1); Anal. Calcd for
C.sub.28H.sub.31N.sub.3O.sub.7.3/4H.sub.2O: C, 62.85; H, 6.12; N,
7.85. Found: C, 62.77; H, 5.95N, 7.79.
Example 213
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]allylamino]etho-
xy]benzoic acid
[1253] ##STR1883##
[1254] To a stirred mixture of ethyl
4-(2-N-trifluoroacetylaminoethoxy)-3-methoxy benzoate (3.5 g, 10.4
mmol) and K.sub.2CO.sub.3 (2.3 g, 16.4 mmol) in DMF (20 mL) was
added allyl bromide (14.2 mL, 16.5 mmol), and the resulting mixture
was stirred for 45 min at 65.degree. C. After cooling, water was
added to the mixture and extracted with EtOAc. The extract was
washed with brine, dried over Na.sub.2SO.sub.4, and evaporated in
vacuo. The residue was dissolved in THF-MeOH--H.sub.2O (1:1:1,
v/v/v) (30 mL) and added K.sub.2CO.sub.3 (2.3 g, 16.4 mmol). The
resulting mixture was stirred for 16 hr at room temp. The mixture
was diluted with EtOAc, washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was chromatographed
on silica-gel with CHCl.sub.3:MeOH (95:5 to 95:5, v/v) as eluent to
give 2.9 g (100%) ethyl 4-(2-allylaminoethoxy)-3-methoxybenzoate as
a pale-yellow oil. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 1.39
(t, 3H, J=7.3 Hz), 3.07 (t, 2H, J=5.3 Hz), 3.34 (d, 2H, J=5.9 Hz),
3.91 (s, 3H), 4.18 (t, 2H, J=5.4 Hz), 4.35 (dd, 2H, J=7.3 Hz, 14.1
Hz), 5.12 (d, 1H, J=10.3 Hz), 5.22 (dd, 1H, J=1.5 Hz, 17.1 Hz),
5.92 (m, 2H), 6.90 (d, 1H, J=8.3 Hz), 7.55 (d, 1H, J=1.5 Hz), 7.65
(dd, 1H, J=2.0 Hz, 8.3 Hz); MS (FAB) m/z 278, 280 (M+H).sup.+.
[1255] To a stirred mixture of ethyl
4-(2-allylaminoethoxy)-3-methoxy benzoate (578 mg, 2.1 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (650 mg,
2.1 mmol), HOBt (420 mg, 3.11 mmol), and DMAP (catalytic amount) in
DMF (4 mL) was added EDC (596 mg, 3.11 mmol) at room temp. The
resulting mixture was stirred for a further 18 hr at room temp. The
mixture was poured into 1N HCl and extracted with EtOAc. The
extract was washed with brine, dried over Na.sub.2SO.sub.4, and
evaporated in vacuo. The residue was chromatographed on silica-gel
with CHCl.sub.3:EtOAc (95:5 to 1:1, v/v) as eluent to give 1 g
(84%)
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]ally-
lamino]ethoxy]benzoic acid as a pale-yellow gum. .sup.1H-NMR
(CDCl.sub.3, 400 Hz) .delta. 1.39 (t, 3H, J=7.3 Hz), 2.29 (s, 3H),
3.58 and 3.63 (s, total 3H), 3.70-3.77 (m, 2H), 3.83 and 3.87 (s,
3H), 4.05-4.13 (m, 2H), 4.25 (m, 1H), 4.36 (q, 1H, J=7.0 Hz),
5.04-5.22 (m, 2H), 5.73 (m, 1H), 6.32 and 6.47 (s, 1H), 6.69-6.85
(m, 2H), 7.12 (m, 2H), 7.23 (m, 2H), 7.50-7.65 (m, 2H), 8.05 (d,
1H, J=7.8 Hz); MS (FAB) m/z 576 (M+H).sup.+.
[1256] A mixture of
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]ally-
lamino]ethoxy]benzoic acid (50 mg, 0.09 mmol) in THF-MeOH (1:1,
v/v) (2 mL) and 1N NaOH (0.135 mL, 0.135 mmol was stirred for 15 hr
at room temp and 3 hr at 50.degree. C. The mixture was poured into
ice-1N HCl. Solid was collected, washed with water, and air-died.
The crude solid was recrystallized from CHCl.sub.3-n-hexane to give
38 mg (77%) 261 as a white crystalline material. mp 125-130.degree.
C.; IR (KBr), 3319, 2939, 1687, 1647, 1601, 1535, 1456, 1417, 1269,
1223, 1034, 760 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6, 400 MHz)
.delta. 2.29 (s, 3H), 3.68 (s, 2H), 3.75-3.85 (m, 8H), 4.05 (br,
1H), 4.19 (m, 3H), 5.10-5.25 (m, 2H), 5.65-5.90 (m, 1H), 6.75 (m,
1H), 6.85 (s, 1H), 6.92 (m, 1H), 7.02-7.20 (m, 3H), 7.48 (d, 1H,
J=10.2 Hz), 7.56 (m, 1H), 7.79 (d, 1H, J=6.8 Hz), 8.01 (m, 1H),
8.46 (s, 1H), 8.56 (d, 1H, J=4.4 Hz), 12.7 (br, 1H); MS (FAB) m/z
548 (M+H).sup.+; Anal. calcd. for
C.sub.30H.sub.33N.sub.3O.sub.7.0.5H.sub.2O, C, 64.74; H, 6.16; N,
7.55. Found, C, 64.72; H, 6.07; N, 7.55.
Example 214
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-mo-
rpholino)ethylamino]ethoxy]benzoic acid
[1257] ##STR1884##
[1258] To a stirred solution of
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]ally-
lamino]ethoxy]benzoic acid (950 mg, 1.65 mmol) in THF:H.sub.2O (7
mL) was added N-methylmorpholine-N-oxide (579 mg, 4.95 mmol) and
osmium tetroxide (0.2M solution in water) (0.413 mL, 0.08 mmol).
The resulting mixture was stirred for 3 hr at room temp. Sat.
NaHSO.sub.3 was added to the mixture, and the mixture was filtered
through Celite. The filtrate was extracted with EtOAc. The extract
was washed with brine, dried over MgSO.sub.4, and evaporated in
vacuo. The residue was dissolved in MeOH-THF--H.sub.2O (1:1:1, v/v)
(12 mL) and added sodium periodate (318 mg, 1.5 mmol). The
resulting mixture was stirred at an ambient temp for 1 hr. The
mixture was diluted with EtOAc, washed with brine, and dried over
MgSO.sub.4. Solvent was evaporated in vacuo to afford 862 mg (90%)
ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-f-
ormylmethylamino]ethoxy]benzoate as a pale-yellow gum. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 1.39 (t, 3H, J=7.3 Hz), 2.29 (s, 3H),
3.31-3.95 (m, 11H), 4.10-4.42 (m, 5H), 6.51-6.82 (m, 3H), 7.10-7.25
(m, 3H), 7.50 (m, 2H), 7.60 (m, 1H), 8.10 (m, 1H), 9.50 (m, 1H); MS
(FAB) m/z 578 (M+H).sup.+.
[1259] To a stirred mixture of ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-f-
ormylmethylamino]ethoxy]benzoate (265 mg, 0.46 mmol), morpholine
(0.40 mL, 4.59 mmol), and AcOH (0.263 mL, 4.6 mmol) in EtOH (3 mL)
was added NaBH.sub.3CN (288 mg, 4.6 mmol) at room temp. The
resulting mixture was stirred for 15 hr at room temp and the
mixture was diluted with EtOAc and added sat. NaHCO.sub.3 at
0.degree. C. The resulting mixture was stirred for 0.5 hr at
0.degree. C. The mixture was extracted with EtOAc. The extract was
washed with brine, dried over MgSO.sub.4, and evaporated in vacuo.
The residue was chromatographed on silica-gel with CHCl.sub.3:MeOH
(95:5, v/v) as eluent to give 213 mg (71%) ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-m-
orpholino)ethylamino]ethoxy]benzoate as an oil. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 1.28 (t, 3H, J=7.0 Hz), 2.31 (s, 3H),
2.48 (brs, 4H), 2.52 (m, 2H), 3.60-3.91 (m, 16H), 4.11 and 4.28 (m,
total 2H), 4.39 (q, 2H, J=7.0 Hz), 6.70-6.85 (m, 4H), 7.15 (m, 2H),
7.50-7.63 (m, 3H), 8.08 (d, 1H, J=8.0 Hz); MS (FAB) m/z 649
(M+H).sup.+.
[1260] A mixture of ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-(2-morpholino)ethylamino]ethoxy]benzoate (265 mg, 0.46
mmol) in THF (4 mL) and 1N NaOH (0.984 mL) was stirred at
50.degree. C. for 15 hr. The pH of the mixture was adjusted to 7.4
by the addition of 1N HCl, and extracted with CHCl.sub.3:MeOH (9:1,
v/v). The extract was washed with brine, dried over MgSO.sub.4, and
evaporated in vacuo. The residue was crystallized with Et.sub.2O to
give 160 mg (78%) 262 as a white crystalline material. mp
125-130.degree. C.; IR (KBr), 3346, 2956, 2937, 1705, 1622, 1599,
1537, 1456, 1417, 1299, 1114, 1032, 752 cm.sup.-1; .sup.1H-NMR
(CD.sub.3OD, 400 MHz) .delta. 2.29 (s, 3H), 2.49-2.64 (m, 6H),
3.65-3.85 (m, 16H), 4.13 (m, 1H), 4.26 (m, 1H), 6.78-7.04 (m, 4H),
7.18 (m, 2H), 7.55-7.64 (m, 3H), 7.99 (m, 2H); MS (FAB) m/z 621
(M+H).sup.+; Anal. Calcd. for
C.sub.33H.sub.40N.sub.4O.sub.8.2.5H.sub.2O, C, 59.54; H, 6.81; N,
8.42. Found, C, 59.71; H, 6.35; N, 7.98.
Example 215
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-[2-[-
4-methyl-1-piperazinyl]ethylamino]ethoxy]benzoic acid
[1261] ##STR1885##
[1262] To a stirred mixture of ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-f-
ormylmethylamino]ethoxy]benzoate (242 mg, 0.42 mmol),
N-methylpiperazine (0.465 mL, 4.2 mmol), and AcOH (0.240 mL, 4.2
mmol) in EtOH (3 mL) was added NaBH.sub.3CN (263 mg, 4.2 mmol) at
room temp. The resulting mixture was stirred for 15 hr at room
temp. The mixture was diluted with EtOAc and added sat. NaHCO.sub.3
at 0.degree. C. The resulting mixture was stirred for 0.5 hr at
0.degree. C. The mixture was extracted with EtOAc. The extract was
washed with brine, dried over MgSO.sub.4, and evaporated in vacuo.
The residue was chromatographed on silica-gel with CHCl.sub.3:MeOH
(95:5, v/v) as eluent to give 195 mg (70%) ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-[2--
(4-methyl-1-piperazinyl]ethylamino]ethoxy]benzoate as an oil.
.sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 1.23 (t, 3H, J=7.0 Hz),
2.25 (s, 3H), 2.29 (s, 3H), 2.50 (br m, 12H), 3.44-3.85 (m, 12H),
4.10 (br, 1H), 4.22 (br, 1H), 4.35 (m, 2H), 6.70-6.85 (m, 3H), 6.98
(s, 1H), 7.10 (m, 1H), 7.20 (m, 2H), 7.40 (m, 1H), 7.60-7.70 (m,
3H), 8.05 (d, 1H, J=7.8 Hz); MS (FAB) m/z 662 (M+H).sup.+.
[1263] A mixture of ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-[2--
(4-methyl-1-piperazinyl]ethylamino]ethoxy]benzoate (195 mg, 0.30
mmol) in THF:MeOH (4:1, v/v) (5 mL) and 1N NaOH (0.885 mL) was
stirred at 50.degree. C. for 15 hr. The pH of the mixture was
adjusted to 7.4 by the addition of 1N HCl, and extracted with
CHCl.sub.3:MeOH (9:1, v/v). The extract was washed with brine,
dried over MgSO.sub.4, and evaporated in vacuo. The residue was
crystallized with Et.sub.2O to give 141 mg (75%) 263 as a white
crystalline material. mp 155-160.degree. C.; IR (KBr), 2937, 1537,
783 cm.sup.-1; .sup.1H-NMR (CD.sub.3OD, 400 MHz) .delta. 2.29 (s,
3H), 2.49-2.80 (m, 15H), 3.60-3.85 (m, 9H), 3.92 (s, 1H), 4.12 (m,
1H), 4.25 (m, 1H), 6.78-7.20 (m, 6H), 7.61 (m, 3H), 8.00 (m, 1H);
MS (FAB) m/z 632 (M).sup.+; Anal. Calcd. for
C.sub.34H.sub.43N.sub.5O.sub.7.2.5H.sub.2O, C, 60.16; H, 7.13; N,
10.32. Found, C, 59.72; H, 6.86; N, 9.97.
Example 216
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-[2-c-
yclopropylamino]ethylamino]ethoxy]benzoic acid
[1264] ##STR1886##
[1265] To a stirred mixture of ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-f-
ormylmethylamino]ethoxy]benzoate (267 mg, 0.46 mmol),
cyclopropylamine (0.32 mL, 4.6 mmol), and AcOH (0.264 mL, 4.6 mmol)
in EtOH (3 mL) was added NaBH.sub.3CN (290 mg, 4.6 mmol) at room
temp. The resulting mixture was stirred for 15 hr at room temp. The
mixture was diluted with EtOAc and added sat NaHCO.sub.3 at
0.degree. C. The resulting mixture was stirred for 0.5 hr at
0.degree. C. The mixture was extracted with EtOAc. The extract was
washed with brine, dried over MgSO.sub.4, and evaporated in vacuo.
The residue was chromatographed on silica-gel with CHCl.sub.3:MeOH
(95:5, v/v) as eluent to give 156 mg (55%) ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-[2--
cyclopropylamino]ethylamino]ethoxy]benzoate as an oil. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 0.35 (m, 4H), 1.22 (br s, 3H), 2.10
(m, 1H), 2.20 (s, 3H), 2.42 (br, 2H), 2.90 (br s, 2H), 3.60-3.80
(m, 10H), 4.10 (br, 1H), 4.22 (br, 1H), 4.33 (br, 2H), 6.72 (m,
3H), 7.05-7.30 (m, 4H), 7.55 (m, 4H), 8.06 (br s, 1H); MS (FAB) m/z
619 (M+H).sup.+.
[1266] A mixture of ethyl
3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-[-
2-cyclopropylamino]ethylamino]ethoxy]benzoate (195 mg, 0.30 mmol))
in THF:MeOH (4:1, v/v) (5 mL) and 1N NaOH (0.756 mL) was stirred at
50.degree. C. for 15 hr. The pH of the mixture was adjusted to 7.4
by the addition of 1N HCl, and extracted with CHCl.sub.3:MeOH (9:1,
v/v). The extract was washed with brine, dried over MgSO.sub.4, and
evaporated in vacuo. The residue was crystallized with Et.sub.2O to
give 57 mg (38%) 3-methoxy-4-[[2-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]-[2-cyclopropylamino]ethylamino]ethoxy]benzoic
acid 264 as a white crystalline material. mp 135-140.degree. C.; IR
(KBr), 3324, 2937, 1535, 1032, 754 cm.sup.-1; .sup.1H-NMR
(CD.sub.3OD, 400 MHz) .delta. 0.50-0.73 (m, 4H), 2.29 (s, 3H), 2.53
(m, 1H), 2.98 (m, 1H), 3.21 (m, 1H), 3.58-3.88 (m, 11H), 3.91 (s,
1H), 4.09 (m, 1H), 4.25 (m, 1H), 6.76-6.92 (m, 3H), 7.01 (m, 1H),
7.18 (m, 2H), 7.60 (m, 3H), 8.00 (d, J=8.3 Hz, 1H); MS (FAB) m/z
591 (M+H).sup.+; Anal. Calcd. for
C.sub.32H.sub.38N.sub.4O.sub.7.3.0H.sub.2O, C, 59.62; H, 6.88; N,
8.69. Found, C, 59.25; H, 6.29; N, 8.29.
Example 217
4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-N-methylamino]-
ethoxy]benzoic acid
[1267] ##STR1887##
3-chloro-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-N-met-
hylaminoethoxy]benzoic acid
[1268] ##STR1888##
[1269] To a stirred cold (0.degree. C.) solution of
2-(N-benzyloxycarbonyl-N-methyl)ethanolamine (3.01 g, 14.4 mmol),
methyl 3-chloro-4-hydroxybenzoate (2.68 g, 14.4 mmol), Ph.sub.3P
(5.65 g, 21.5 mmol) in THF (30 mL) was added diisopropyl
azodicarboxylate (DIAD) (4.25 mL, 21.6 mmol), and the resulting
mixture was heated under reflux overnight. The solution was
evaporated off and the residue was purified by column
chromatography on silica-gel with CHCl.sub.3 as eluent to give 3.90
g (72%) methyl
3-chloro-4-[2-(N-benzyloxycarbonyl-N-methylamino)ethoxy]benzoate as
a pale yellow solid. .sup.1H-NMR (CDCl.sub.3) .delta. 3.15 (s, 3H),
3.74-3.76 (m, 2H), 3.89 (s, 3H), 4.17-4.27 (m, 2H), 5.14 (s, 2H),
6.81-6.94 (m, 1H), 7.33-7.36 (m, 5H), 7.85-7.92 (m, 1H), 8.05 (bs,
1H).
[1270] A solution of methyl
3-chloro-4-[2-(N-benzyloxycarbonyl-N-methylamino)ethoxy]benzoate
(3.90 g, 10.3 mmol) in EtOAc--AcOH (40 mL, 1:1, v/v) was
hydrogenated over 5% Pd--C (1.95 g, 50 wt %) at 3 atm for 2 hr. The
mixture was filtered and the filtrate was washed with sat.
NaHCO.sub.3 and the basic aqueous layer was extracted with
CHCl.sub.3, washed with brine and evaporate to give unseparable
mixture of methyl 3-chloro-4-(N-methylaminoethoxy)benzoate and
methyl 4-[2-(N-methylamino)ethoxy]benzoate the title compound (1.61
g) as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.52-2.52
and 2.53-2.54 (each m, 3H), 2.98-3.00 and 3.03-3.05 (each m, each
2H), 3.88 and 3.99 (each s, each 3H), 4.11-4.14 and 4.18-4.20 (each
m, each 2H), 6.91-6.96 and 7.90-8.05 (series of m, total 7H).
[1271] A mixture of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (392 mg),
a mixture of methyl 3-chloro-4-[2-(N-methylamino)ethoxy]benzoate
and methyl 4-[2-(N-methylamino)ethoxy]benzoate (305 mg), EDC
(hydrochloride) (354 mg), HOBt (250 mg), and DMAP (250 mg) in DMF
(8 mL) was stirred at room temp for 6 hr. The mixture was diluted
with EtOAc, washed with 0.5 N HCl, brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with 1% MeOH in CHCl.sub.3 as eluent
to give a mixture of methyl
3-chloro-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-N-me-
thylaminoethoxy]benzoate and methyl
4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-N-methylamino-
ethoxy]benzoate (550 mg) as a brown amorphous solid.
[1272] To a stirred solution of this mixture (550 mg) of methyl
3-chloro-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-N-me-
thylaminoethoxy]benzoate and methyl
4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]-N-methylamino-
ethoxy]benzoate in THF-MeOH (20 mL, 1:1, v/v) was added 0.5 N NaOH
(10 mL), and the resulting mixture was heated under reflux for 6
hr. The mixture was poured into ice-1 N HCl, and the solid was
collected. The crude solid was purified by preparative TLC with 10%
MeOH in CHCl.sub.3 as eluent to give 265 (56 mg, as a white
amorphous solid) and 266 (88 mg, as a brown amorphous solid).
Example 218
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]eth-
oxy]benzoic acid
[1273] ##STR1889##
3-chloro-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoic acid
[1274] ##STR1890##
[1275] A mixture of methyl
4-[2-(N-methyl-2-amino)ethoxy]-3-chlorobenzoate (292 mg, 1.2 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (377 mg,
1.2 mmol), EDC (345 mg, 1.8 mmol), HOBt (243 mg, 1.8 mmol), and
DMAP (29 mg, 0.24 mmol) in DMF (2.7 mL) was stirred for 6 hr at
room temp. The mixture was poured into ice-1N HCl and extracted
with EtOAc. The extract was washed with brine, dried over
MgSO.sub.4, and evaporated in vacuo. The residue was
chromatographed on silica-gel with CHCl.sub.3:EtOAc (95:5 to 0:100,
v/v) as eluent to give unseparable mixture (489 mg) of methyl
3-chloro-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methy-
lamino]ethoxy]benzoate and methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]methylamino]ethoxy]benzoate as pale-yellow oil.
[1276] A mixture (480 mg as mixture) of methyl
3-chloro-4-[[2-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]methylamino]ethoxy]benzoate and methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]et-
hoxy]benzoate in THF-MeOH (4 mL, 1:1, v/v) was stirred at
50.degree. C. for 15 hr. The mixture was poured into ice-1N HCl.
The solid was collected, washed with water, and air-dried. The
crude solid was purified by preparative TLC CHCl.sub.3:MeOH (93:7,
v/v) as eluent to afford 267 (180 mg, 2 steps 31% as a crystalline
material) and 268 (280 mg, 2 steps 44% as a crystalline material).
267: mp 145-150.degree. C.; .sup.1H-NMR (DMSO-d.sub.6, 400 MHz)
.delta. 2.31 (s, 3H), 3.05 and 3.19 (s, 3H), 3.35 and 3.38 (s, 3H),
3.72-3.85 (m, 7H), 4.09 and 4.23 (m, total 2H), 6.79-7.20 (m, 7H),
7.60 (m, 1H), 7.86-8.09 (m, 3H); MS (FAB) m/z 493 (M+H).sup.+;
Anal. calcd. for C.sub.27H.sub.29N.sub.3O.sub.6.1.75H.sub.2O, C,
62.00; H, 6.26; N, 8.03. Found, C, 62.16; H, 5.88; N, 7.82. 268: mp
145-150.degree. C.; .sup.1H-NMR (DMSO-d.sub.6, 400 MHz) .delta.
2.29 (s, 3H), 3.06 and 3.26 (s, 3H), 3.31 and 3.35 (s, 3H),
3.85-3.94 (m, 4H), 4.18 and 4.32 (m, total 2H), 6.75-6.85 (m, 2H),
6.99-7.20 (m, 4H), 7.59 (m, 1H), 7.90-8.02 (m, 3H); MS (FAB) m/z
526 (M+H).sup.+; Anal. calcd. for
C.sub.27H.sub.28ClN.sub.3O.sub.6.2.0H.sub.2O, C, 57.70; H, 5.74; N,
7.48. Found, C, 57.99; H, 5.53; N, 7.07.
Example 219
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-methylamino]--
1-propyl]benzoic acid
[1277] ##STR1891##
[1278] To a stirred cold (minus 78.degree. C.) solution of triethyl
4-phosphonomethylbenzoate (1.22 g, 4.05 mmol) in THF (10 mL) was
added NaHMDS (1.0 M in THF) (4.0 mL, 4.0 mmol), and the resulting
mixture was stirred for 1 hr at the same temp. A solution of
2-(N-benzyloxycarbonyl-N-methylamino)acetaldehyde (700 mg, 3.38
mmol) in THF (5 mL) was slowly added to this solution at that temp,
and the mixture was allowed to warm to room temp for over 2 hr with
stirring. The solution was quenched by the addition of sat.
NH.sub.4Cl (100 mL), and extracted with EtOAc. The extract was
washed with brine (200 mL), dried over MgSO.sub.4, and evaporated.
The residue was chromatographed on silica gel with CHCl.sub.3-EtOAc
(20:1, v/v) as eluent to give 810 mg (68%) ethyl
(E)-4-[3-(N-benzyloxycarbonyl-N-methylamino)-1-propenylbenzoate as
a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.40 (t, J=7.3 Hz,
3H), 2.95 (m, 2H), 4.09 (m, 2H), 4.35-4.40 (m, 2H), 5.17 (s, 2H),
6.26-6.64 (series of m, 2H), 7.36 (m, 7H), 7.99 (d, J=8.3 Hz,
2H).
[1279] A stirred solution of ethyl
(E)-4-[3-(N-benzyloxycarbonyl-N-methylamino)-1-propenyl benzoate
(810 mg, 2.29 mmol) in EtOH--AcOH (10:1, v/v, 22 mL) was
hydrogenated over 5% Pd--C (1 g) for 3 days. The mixture was
filtered and the filtrate was evaporated. The residue was made
basic with sat. NaHCO.sub.3 and extracted with CHCl.sub.3. The
extract was dried over Na.sub.2CO.sub.3 and evaporated to give 438
mg (86%) ethyl 4-(3-methylamino-1-propyl)benzoate as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.39 (t, J=7.3 Hz, 3H), 1.82 (m,
2H), 2.43 (s, 3H), 2.61 (t, J=7.3 Hz, 2H), 2.72 (t, J=7.3 Hz, 2H),
3.33 (br s, 1H), 4.36 (q, J=7.3 Hz, 2H), 7.25 (d, J=8.3 Hz, 2H),
7.96 (d, J=8.3 Hz, 2H).
[1280] To a stirred solution of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (456 mg,
1.45 mmol) and ethyl 4-(3-methylamino-1-propyl)benzoate (220 mg,
1.45 mmol) were added EDC.HCl (417 mg, 2.16 mmol), HOBt (cat.), and
DMAP (catalytic amount) in DMF (10 mL), and the resulting mixture
was stirred overnight. The mixture was diluted with EtOAc (300 mL),
washed with brine, dried over MgSO.sub.4, and evaporated. The
residue was chromatographed on silica-gel with CHCl.sub.3-EtOH
(10:1) to give 503 mg (71%) ethyl
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl-N-methylamino]--
1-propyl]benzoate as a yellow oil. MS (FAB) m/z 518
(M+H).sup.+.
[1281] To a stirred solution of ethyl
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl-N-methylamino]-1-propyl]benzoate (500 mg, 0.966 mmol) in THF
(8 mL) was added 0.25 N NaOH (8 mL), and the mixture was heated
under reflux overnight. The resulting solution was poured into
ice-1 N HCl (100 mL) and the solid was collected with suction. The
solid was dissolved in CHCl.sub.3 (100 mL) and dried over
MgSO.sub.4. After removal of the solvent, the residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1 to 5:1,
v/v) to give 131 mg (28%)
4-[3-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl-N-methylamino]--
1-propyl]benzoic acid 269 as a pale yellow amorphous solid.
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.68-1.80 (m, 2H), 2.24 (s, 3H),
2.57 (m, 2H), 2.81 and 2.97 (s, each, total 3H), 3.33 (m, 2H),
3.57-3.61 (m, 2H), 3.84 (s, 3H), 6.71 (dd, J=29.8, 8.3 Hz, 1H),
6.87 (d, J=11.2 Hz, 1H), 6.93 (t, J=7.3 Hz, 1H), 7.15 (m, 2H), 7.28
(m, 2H), 7.79 (d, J=8.3 Hz, 1H), 7.84-7.87 (m, 2H), 8.02 (d, J=8.3
Hz, 1H), 8.49 (d, J=7.3 Hz, 1H), 8.58 (d, J=4.9 Hz, 1H); MS (FAB)
m/z 490 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.5.1/2H.sub.2O: C, 67.45; H, 6.47; N,
8.43. Found: C, 67.27; H, 6.51; N, 8.02.
Example 220
4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-methylamino]ethoxy]ben-
zoic acid
[1282] ##STR1892##
3-chloro-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-methylamino]e-
thoxy]benzoic acid
[1283] ##STR1893##
[1284] A solution of pentafluorophenyl
4-[N'-(2-methylphenyl)ureido]phenylacetate (562 mg, 1.29 mmol), a
mixture (304 mg) of methyl
3-chloro-4-[2-(N-methylamino)ethoxy]benzoate and methyl
4-[2-(N-methyl amino)ethoxy]benzoate and Et.sub.3N (260 mL) in DMF
(8 mL) was stirred at room temp for 4 hr. The mixture was diluted
with EtOAc, washed with 0.5 N HCl, brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as
eluent to give a mixture (670 mg) of methyl
3-chloro-4-[[2-[4-[N'-(2-methylphenyl)ureido)phenylacetyl]-N-methylamino)-
ethoxy]benzoate and methyl
4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]-N-methylamino)ethoxy]be-
nzoate as an oil.
[1285] To a stirred suspension of this mixture (670 mg) in THF-MeOH
(20 mL, 1:1, v/v) was added 0.5 N NaOH (10 mL) and the resulting
mixture was heated under reflux for 6 hr. The solution was poured
into ice-1 N HCl and the solid was collected. The crude solid was
purified by preparative thin layer chromatography (TLC) with 10%
MeOH in CHCl.sub.3 as eluent to give 73 mg 270 as an amorphous
solid and 110 mg 271 as a white amorphous solid. 270 MS (FAB) m/z
462 (M.sup.++1). 271 MS (FAB) m/z 496 (M.sup.++1).
Example 221
(S)-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-prop-
oxy]benzoic acid
[1286] ##STR1894##
[1287] To a cooled (0.degree. C.) solution of
(S)-2-amino-1-propanol (2.08 g, 27.7 mmol) and Et.sub.3N (4.63 mL,
33.2 mmol) in DMF-H.sub.2O (40 mL, 1:1, v/v) was added (Boc).sub.2O
(6.36 mL, 27.7 mmol), and the resulting solution was stirred at
room temp for 2 days. H.sub.2O was added to the mixture and
extracted with EtOAc. The extract was washed with brine and dried
over Na.sub.2SO.sub.4. The solvent was evaporated to give 4.24 g
(87%) (S)-2-(N-tert-butoxycarbonylamino)-1-propanol as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.14 (d, 3H, J=6.8 Hz), 1.45
(s, 9H), 3.51-3.52 (m, 1H), 3.63-3.66 (m, 1H), 3.77 (m, 1H), 4.62
(m, 1H).
[1288] To a cooled (0.degree. C.) solution of
(S)-2-(N-tert-butoxycarbonylamino)-1-propanol (1.02 g, 5.82 mmol),
methyl 4-hydroxybenzoate (0.89 g, 5.85 mmol), and Ph.sub.3P (1.98
g, 7.55 mmol) in THF (20 mL) was added diisopropyl azodicarboxylate
(DIAD) (1.49 mL, 7.57 mmol), and the resulting mixture was heated
under reflux overnight. The solution was evaporated and the residue
was dissolved in CH.sub.2Cl.sub.2 (20 mL) and TFA (10 mL). The
mixture was stirred at room temp for 1.5 hr. The solution was
concentrated in vacuo and the residue was treated with sat.
NaHCO.sub.3. The mixture was extracted with CHCl.sub.3, washed with
brine, dried over Na.sub.2SO.sub.4 and evaporated. The residue was
purified by column chromatography on silica-gel with CHC.sub.3:MeOH
(50:1, v/v) to give 480 mg (2 steps 39%) methyl
(S)-4-(2-amino-1-propoxy)benzoate as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.19 (d, 3H, J=6.4 Hz), 3.35-3.39 (m, 1H),
3.72-3.76 (m, 1H), 3.89 (s, 3H), 3.90-3.94 (m, 1H), 6.92 (d, 2H,
J=8.8 Hz), 7.99 (d, 2H, J=8.8 Hz).
[1289] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (505 mg, 1.05
mmol), methyl (S)-4-(2-amino-1-propoxy)benzoate (220 mg, 1.05
mmol), and Et.sub.3N (0.220 mL, 1.58 mmol) in DMF (8 mL) was
stirred at room temp for 3 hr. The mixture was diluted with EtOAc,
washed with 0.5 N HCl, brine, and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was recrystallized from
MeOH--CHCl.sub.3-n-hexane to give 290 mg (55%) methyl
(S)-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-pro-
poxy]benzoate as a white crystalline powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.18 (d, 3H, J=6.8 Hz), 2.24 (s, 3H), 3.36
(s, 2H), 3.80 (s, 3H), 3.82 (s, 3H), 3.93-4.03 (m, 2H), 4.09-4.14
(m, 1H), 6.75-8.57 (series of m, total 13H).
[1290] To a stirred solution of methyl
(S)-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetylamino]-1-propoxy]benzoate (290 mg, 0.57 mmol) in THF-MeOH (20
mL, 1:1, v/v) was added 0.5 N NaOH (20 mL) and the solution was
heated under reflux for 2 hr. The mixture was poured into ice-1 N
HCl and extracted with CHCl.sub.3-MeOH (10:1, v/v). The extract was
washed with brine, dried over Na.sub.2SO.sub.4 and evaporated. The
residue was recrystallized from MeOH--CHCl.sub.3-n-hexane to give
158 mg (56%) 272 as a white crystalline powder. mp 198-201.degree.
C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.18 (d, 3H, J=6.3 Hz), 2.24
(s, 3H), 3.36 (s, 2H), 3.82 (s, 3H), 3.87-4.10 (m, 2H), 4.10-4.16
(m, 1H), 6.75-6.78 (m, 1H), 6.92-7.02 (m, 4H), 7.11-7.18 (m, 2H),
7.78-7.80 (m, 1H), 7.86-7.89 (m, 2H), 7.98-8.00 (m, 1H), 8.12-8.14
(m, 1H), 8.46 (s, 1H), 8.55 (s, 1H), 12.62 (bs, 1H); MS (FAB) m/z
492 (M.sup.++1); Anal. Calcd for
C.sub.27H.sub.29N.sub.3O.sub.6.1/2H.sub.2O: C, 64.79; H, 6.04; N,
8.21. Found: C, 64.36; H, 5.85; N, 8.21.
Example 222
(S)-4-[2-[4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-propoxy]benzoi-
c acid
[1291] ##STR1895##
[1292] A mixture of pentafluorophenyl
4-[N'-(2-methylphenyl)ureido]phenylacetate (560 mg, 1.24 mmol),
methyl (S)-4-(2-amino-1-propoxy)benzoate (260 mg, 1.24 mmol),
Et.sub.3N (0.260 mL, 1.87 mmol) in DMF (8 mL) was stirred at room
temp for 3 hr. The mixture was diluted with EtOAc and the solution
was washed with 0.5 N HCl, brine, and dried over Na.sub.2SO.sub.4.
After removal of the solvent, the residue was purified by
recrystallization from MeOH--CHCl.sub.3-n-hexane to give 210 mg
(36%)
(S)-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-pro-
poxy]benzoic acid as a white crystalline powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.17 (d, 3H, J=6.8 Hz), 2.24 (s, 3H), 3.32
(s, 2H), 3.81 (s, 3H), 3.92-4.03 (m, 2H), 4.08-4.15 (m, 1H),
6.92-6.95 (m, 1H), 7.04-7.06 (m, 2H), 7.12-7.18 (m, 4H), 7.35-7.39
(m, 2H), 7.83-7.85 (m, 1H), 7.89-7.92 (m, 3H), 8.12-8.14 (m, 1H),
8.97 (s, 1H).
[1293] To a stirred solution of methyl
(S)-4-[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl
amino]-1-propoxy]benzoate (200 mg, 0.42 mmol) in THF-MeOH (10 mL,
1:1, v/v) was added 0.5 N NaOH (10 mL), and the mixture was heated
under reflux for 2 hr. The mixture was poured into ice-1 N HCl, and
the solid was collected. The crude solid was recrystallized from
MeOH--CHCl.sub.3-n-hexane to give 68 mg (34%) 273 as a white
crystalline powder. mp 262-265.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.17 (d, 3H, J=6.8 Hz), 2.24 (s, 3H), 3.32
(s, 2H), 3.91-4.02 (m, 2H), 4.09-4.15 (m, 1H), 6.92-6.96 (m, 1H),
7.01-7.03 (m, 2H), 7.12-7.20 (m, 4H), 7.36-7.40 (m, 2H), 7.83-7.95
(m, 4H), 8.12-8.14 (m, 1H), 8.99 (s, 1H), 12.63 (bs, 1H); MS (FAB)
m/z 462 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.27N.sub.3O.sub.5.1/4H.sub.2O: C, 67.01; H, 5.95; N,
9.02. Found: C, 67.13; H, 5.90; N, 9.02.
Example 223
(S)-3-chloro-4-[2-[4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-propo-
xy]benzoic acid
[1294] ##STR1896##
[1295] To a cooled (0.degree. C.) solution of
(S)-2-(N-tert-butoxycarbonylamino)-1-propanol (1.05 g, 5.99 mmol),
methyl 3-chloro-4-hydroxybenzoate (1.12 g, 6.00 mmol), and
Ph.sub.3P (2.36 g, 9.00 mmol) in THF (20 mL) was added diisopropyl
azodicarboxylate (DIAD) (1.77 mL, 8.99 mmol), and the resulting
mixture was heated under reflux for 2 days. The solution was
evaporated off and the residue was dissolved in CH.sub.2Cl.sub.2
(20 mL) and TFA (10 mL). The resulting mixture was stirred at room
temp for 1.5 hr. The solution was concentrated in vacuo, and the
residue was dissolved in CHCl.sub.3. The mixture was extracted with
H.sub.2O, and the aqueous layer was made basic by the addition of
sat. NaHCO.sub.3. This basic aqueous layer was extracted with
CHCl.sub.3. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated to give 660 mg (2 steps, 32%)
methyl (S)-3-chloro-4-(2-amino-1-propoxy)benzoate as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.21 (d, 3H, J=6.4 Hz),
3.41-3.48 (m, 1H), 3.77-3.81 (m, 1H), 3.89 (s, 3H), 3.98-4.01 (m,
1H), 6.91-6.94 (m, 1H), 7.90-7.93 (m, 1H), 8.05-8.06 (m, 1H).
[1296] A mixture of pentafluorophenyl
4-[N'-(2-methylphenyl)ureido]phenylacetate (508 mg, 1.13 mmol),
methyl (S)-3-chloro-4-(2-amino-1-propoxy)benzoate (275 mg, 1.13
mmol) and Et.sub.3N (0.240 ml, 1.72 mmol) in DMF (10 mL) was
stirred at room temp overnight. The mixture was diluted with EtOAc,
and the solution was washed with 0.5 N HCl, sat. NaHCO.sub.3,
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residue was
recrystallized from MeOH--CHCl.sub.3-n-hexane to give 240 mg (42%)
methyl
(S)-3-chloro-4-[2-[4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-prop-
oxy]benzoate as a white crystalline powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.21 (d, 3H, J=6.4 Hz), 2.25 (s, 3H), 3.33
(s, 2H), 3.82 (s, 3H), 4.04-4.14 (m, 3H), 6.90-6.94 (m, 1H),
7.11-7.16 (m, 4H), 7.29-7.38 (m, 3H), 7.83-7.94 (m, 3H), 8.13-8.17
(m, 2H), 9.34 (s, 1H); MS (FAB) m/z 510 (M.sup.+).
[1297] To a stirred solution of methyl
(S)-3-chloro-4-[2-[4-[N'-(2-methylphenyl)ureido]phenylacetylamino]-1-prop-
oxy]benzoate (240 mg, 0.47 mmol) in THF-MeOH (10 mL, 1:1, v/v) was
added 0.5 N NaOH (10 mL), and the resulting mixture was heated
under reflux overnight. The mixture was poured into ice-1 N HCl and
the solid was collected. The crude solid was recrystallized from
MeOH--CHCl.sub.3-n-hexane to give 98 mg (42%) 274 as a white
crystalline powder. mp 228-231.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.20 (d, 3H, J=6.3 Hz), 2.24 (s, 3H), 3.34
(s, 2H), 4.02-4.18 (m, 3H), 6.92-6.95 (m, 1H), 7.12-7.42 (series of
m, total 7H), 7.82-8.18 (series of m, total 5H), 9.12 (s, 1H); MS
(FAB) m/z 496 (M.sup.+), 497 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.26ClN.sub.3O.sub.5.1/2H.sub.2O: C, 61.84; H, 5.39;
Cl, 7.02; N, 8.32. Found: C, 61.76; H, 5.25; Cl, 7.09; N, 8.25.
Example 224
(S)-3-chloro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamin-
o]-1-propoxy]benzoic acid
[1298] ##STR1897##
[1299] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (513 mg, 1.07
mmol), methyl (S)-3-chloro-4-(2-amino-1-propoxy)benzoate (260 mg,
1.07 mmol) and Et.sub.3N (220 ul, 1.58 mmol) in DMF (10 mL) was
stirred at room temp overnight The mixture was diluted with EtOAc
and the solution was washed with sat. NaHCO.sub.3, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was recrystallized
from MeOH--CHCl.sub.3-EtOAc-n-hexane to give 400 mg (69%) methyl
(S)-3-chloro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylami-
no]-1-propoxy]benzoate as pale brown crystalline powder.
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.21 (d, 3H, J=6.4 Hz), 2.24 (s,
3H), 3.37 (s, 2H), 3.82 (s, 3H), 3.83 (s, 3H), 4.04-4.12 (m, 3H),
6.75-6.77 (m, 1H), 6.91-6.95 (m, 2H), 7.11-7.17 (m, 2H), 7.29-7.31
(m, 1H), 7.78-7.99 (m, 4H), 8.12-8.13 (m, 1H), 8.46 (s, 1H), 8.55
(s, 1H).
[1300] To a stirred solution of methyl
(S)-3-chloro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylami-
no]-1-propoxy]benzoate (400 mg, 0.74 mmol) in THF-MeOH (20 mL, 1:1,
v/v) was added 0.5 N NaOH (20 mL), and the resulting mixture was
heated under reflux overnight. The mixture was poured into ice-1 N
HCl and the solid was collected. The crude solid was recrystallized
from MeOH--CHCl.sub.3-Et.sub.2O to give 200 mg (51%) 275 as a pale
brown crystalline powder. mp 198-201.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.21 (d, 3H, J=6.8 Hz), 2.24 (s, 3H), 3.37
(s, 2H), 3.84 (s, 3H), 4.00-4.15 (m, 3H), 6.76-7.28 (series of m,
total 6H), 7.77-8.14 (series of m, total 5H), 8.46 (s, 1H), 8.56
(s, 1H); MS (FAB) m/z 526 (M.sup.+), 528 (M.sup.++2); Anal. Calcd
for C.sub.17H.sub.28ClN.sub.36O.sub.6.1/4H.sub.2O: C, 61.13; H,
5.42; Cl, 6.68; N, 7.92. Found: C, 60.97; H, 5.48; Cl, 6.86; N,
7.89.
Example 225
3-dimethylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl-
]methylamino]ethoxy]benzoic acid
[1301] ##STR1898##
[1302] To a stirred and cooled (0.degree. C.) solution of
2-(N-Boc-N-methylamino)ethanol (3 g, 17 mmol), methyl
4-hydroxy-3-nitro benzoate (3.38 g, 17 mmol), and Ph.sub.3P (5.4 g,
21 mmol) in THF (20 mL) was added diisopropyl azodicarboxylate
(DIAD) (4 mL, 21 mmol), and the resulting mixture was heated under
reflux for 15 hr. The solution was evaporated off. The residue was
chromatographed on silica-gel with CHCl.sub.3:MeOH (100:0 to 4:1,
v/v) as eluent to give 2.5 g (39%) methyl
4-[2-(NV-methyl-2-amino)ethoxy]-3-nitro benzoate as a pale yellow
oil. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2-82 (s, 3H), 3.50
(t, 2H, J=4.5 Hz), 3.95 (s, 3H), 4.54 (t, 2H, J=4.5 Hz), 7.26 (d,
1H, J=8.8 Hz), 8.25 (d, 1H, J=8.8 Hz), 8.56 (s, 1H; MS (FAB) m/z
255 (M.sup.++1).
[1303] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (992 mg,
3.11 mmol), methyl 4-(N-methyl-2-aminoethoxy)-3-nitro benzoate (800
mg, 3.11 mmol) and 4-DMAP (77 mg, 0.63 mmol), HOBt (640 mg, 4.7
mmol), and EDC (904 mg, 4.7 mmol) in DMF (20 mL) was stirred at
room temp overnight. The mixture was diluted with EtOAc, and the
solution was washed with 0.5 N HCl, sat. NaHCO.sub.3, brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with CHCl.sub.3:EtOAc (95:5 to 0:100,
v/v) as eluent to give 587 mg (34%) methyl
3-nitro-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate as a pale yellow oil. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 2.31 (s, 3H), 3.05 (s, 1H), 3.23 (s,
2H), 3.71 (s, 2H), 3.83 (s, 3H), 3.85 (m, 3H), 3.94 (s, 3H), 4.19
and 4.39 (m, total 2H), 6.80 (m, 2H), 7.05 (m, 1H), 7.22 (m, 3H),
7.62 (d, 1H, J=8.2 Hz), 8.02 (d, 1H, J=8.2 Hz), 8.21 (dd, 1H, J=2.1
Hz, 8.8 Hz, 8.55 (d, 1H, J=2.1 Hz); MS (FAB) m/z 551
(M.sup.++1).
[1304] A mixture of methyl
3-nitro-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]methylamino]ethoxy]benzoate (587 mg, 1.1 mmol) and 5%-Pd--C
(600 mg) in THF-MeOH--AcOH (1:1:1, v/v, 150 mL) was hydrogenated at
45 psi for 18 hr. Insoluble catalyst was removed with suction, and
the filtrate was evaporated in vacuo to afford 555 mg (100%) methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate as a pale yellow gum. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 2.29 (s, 3H), 3.08 (m, 1H), 3.19 (s,
2H), 3.65 (s, 1H), 3.73-3.80 (m, 3H), 3.84 (s, 3H), 3.87 (s, 3H),
4.19 and 4.40 (m, total 2H), 6.70-6.82 (m, 2H), 7.02-7.29 (m, 6H),
7.60 (d, 1H, J=7.8 Hz), 7.92-7.99 (m, 3H); MS (FAB) m/z 521
(M.sup.++1).
[1305] To a stirred solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (555 mg, 1.1 mmol), formaldehyde (10 mL), and
AcOH (0.58 mL, 10 mmol) in MeCN (10 mL) was added NaBH.sub.3CN
(0.67 g, 10 mmol) at room temp, and the resulting mixture was
stirred for 15 hr at the same temp. Sat. NaHCO.sub.3 was added to
the mixture and extracted with CHCl.sub.3. The extract was washed
with brine, dried over MgSO.sub.4, and evaporated in vacuo. The
residue was chromatographed on silica-gel with toluene:acetone (7:3
to 1:1, v/v) as eluent to give 123 mg (21%) methyl
3-dimethylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacety-
l]methylamino]ethoxy]benzoate as an oil. .sup.1H-NMR (CDCl.sub.3,
400 MHz) .delta. 2.30 (s, 3H), 2.70 (s, 3H), 2.75 (s, 3H), 3.05 (s,
1H), 3.18 (s, 2H), 3.61 (s, 3H), 3.70 (s, 1H), 3.80 (m, 3H), 3.86
(s, 3H), 4.07 and 4.22 (m, total 2H), 6.28 (m, 1H), 6.70-6.80 (m,
3H), 7.03 (m, 1H), 7.15-7.25 (m, 4H), 7.46-7.65 (m, 2H), 8.02 (m,
1H); MS (FAB) m/z 548 (M.sup.++1).
[1306] A stirred mixture of methyl
3-dimethylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacety-
l]methylamino]ethoxy]benzoate (123 mg, 0.22 mmol) in THF (15 mL)
and 1N NaOH (0.885 mL, 0.885 mmol) was heated under reflux for 15
hr. The pH of the mixture was adjusted to 5.0 by the addition of 1N
HCl, and extracted with CHCl.sub.3-MeOH (9:1, v/v). The extract was
washed with brine, dried over MgSO.sub.4, and evaporated in vacuo.
The residue was crystallized with Et.sub.2O-n:hexane to give 118 mg
(100%) 276 as a white crystalline material. mp 125-130.degree. C.;
IR (KBr) 3346, 2940, 1620, 1597, 1535, 1456, 1417, 1227, 1039, 754
cm.sup.-1; .sup.1H-NMR (CD.sub.3OD, 400 MHz) .delta. 2.29 (s, 3H),
2.70 (s, 3H), 2.79 (s, 2H), 3.05 (s, 1H), 3.22 (s, 2H), 3.75 (s,
3H), 3.85 (m, 4H), 4.15 and 4.28 (m, 2H), 6.78-7.05 (m, 4H), 7.18
(m, 2H), 7.55-7.70 (m, 3H), 7.98 (m, 1H); MS (FAB) m/z 535
(M.sup.++1); Anal. calcd. for
C.sub.29H.sub.34N.sub.4O.sub.6.2.0H.sub.2O: C, 61.04; H, 6.71; N,
9.82. Found: C, 61.15; H, 6.43; N, 8.94.
Example 226
3-dimethylamino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl-
]methylamino]ethoxy]benzoic acid
[1307] ##STR1899##
[1308] A mixture of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (1 g, 3.11
mmol), methyl 4-[2-(N-methyl-2-amino)ethoxy]-3-nitrobenzoate (800
mg, 3.11 mmol) and 4-DMAP (77 mg, 0.63 mmol), HOBt (640 mg, 4.7
mmol), and EDC (904 mg, 4.7 mmol) in DMF (20 mL) was stirred at
room temp overnight. The mixture was diluted with EtOAc, and the
solution was washed with 0.5 N HCl, sat. NaHCO.sub.3, brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with CHCl.sub.3-EtOAc (95:5 to 0:100,
v/v) as eluent to give 420 mg (19%) methyl
3-nitro-4-[[2-[3-fluoro-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyla-
mino]ethoxy]benzoate as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3,
400 MHz) .delta. 3.04 (s, 1H), 3.24 (s, 2H), 3.72 (s, 1H), 3.85 (s,
3H), 3.90 (m, 3H), 3.93 (s, 3H), 4.16 and 4.39 (2m, 3H), 6.80 (m,
2H), 6.99 (m, 1H), 7.05 (m, 2H), 7.22 (d, 1H, J=8.8 Hz), 7.51 (s,
2H), 8.00 (m, 1H), 8.08 (m, 1H), 8.21 (d, 1H, J=8.6 Hz), 8.51 (s,
1H); MS (FAB) m/z 555 (M.sup.++1).
[1309] A mixture of methyl
3-nitro-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (420 mg, 0.76 mmol) and 5%-Pd--C (1 g) in
THF-MeOH--AcOH (1:1:1, v/v, 150 mL) was hydrogenated at 45 psi for
18 hr. Insoluble catalyst was removed with suction, and the
filtrate was evaporated in vacuo to afford 397 mg (100%) methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate as a pale yellow gum. .sup.1H-NMRv
(CDCl.sub.3, 400 MHz) .delta. 3.05 and 3.13 (s, total 3H), 3.66 (s,
3H), 3.70 (s, 2H), 3.65-3.90 (m, 4H), 3.86 (s, 3H), 4.10 and 4.23
(m, 2H), 6.70-6.83 (m, 3H), 6.98-7.15 (m, 6H), 7.25-7.43 (m, 2H),
7.99 (m, 1H), 8.13 (m, 1H); MS (FAB) m/z 525 (M.sup.++1).
[1310] To a stirred solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (397 mg, 0.76 mmol), formaldehyde (10 mL),
and AcOH (0.43 mL, 7.6 mmol) in MeCN (10 mL) was added NaBH.sub.3CN
(0.48 g, 7.6 mmol) at room temp, and the resulting mixture was
stirred for 15 hr at the same temp. Sat. NaHCO.sub.3 was added to
the mixture and extracted with CHCl.sub.3. The extract was washed
with brine, dried over MgSO.sub.4, and evaporated in vacuo. The
residue was chromatographed on silica-gel with toluene:acetone (7:3
to 1:1, v/v) as eluent to give 123 mg (21%) methyl
3-dimethylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacety-
l]methylamino]ethoxy]benzoate as an oil. .sup.1H-NMR (CDCl.sub.3,
400 MHz) .delta. 2.74 (s, 3H), 2.77 (s, 3H), 3.08 (s, 1H), 3.22 (s,
2H), 3.52 (s, 3H), 3.61 (s, 1H), 3.83 (m, 3H), 3.88 (s, 3H), 4.12
and 4.23 (m, total 2H), 6.68 (s, 1H), 6.78 (m, 2H), 6.98 (m, 2H),
7.10 (m, 1H), 7.55-7.68 (m, 4H), 7.99 (m, 1H), 8.16 (t, 1H, J=8.3
Hz); MS (FAB) m/z 553 (M.sup.++1).
[1311] A stirred mixture of methyl
3-dimethylamino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacety-
l]methylamino]ethoxy]benzoate (61 mg, 0.11 mmol) in THF (15 mL) and
1N NaOH (0.22 mL, 0.22 mmol) was heated under reflux for 15 hr. The
pH of the mixture was adjusted to 5.0 by the addition of 1N HCl,
and extracted with CHCl.sub.3:MeOH (9:1, v/v). The extract was
washed with brine, MeOH:acetone (93:7, v/v) as eluent to give 37 mg
(63%) 277 as a white crystalline material. mp 120-125.degree. C.;
.sup.1H-NMR (CD.sub.3OD, 400 MHz) .delta. 2.60 (s, 4H), 2.78 (s,
2H), 3.06 (s, 1H), 3.22 (s, 2H), 3.75 (s, 3H), 3.85-3.92 (m, 4H),
4.17 and 4.29 (m, total 2H), 6.80-7.12 (m, 6H), 7.61-7.70 (m, 2H),
8.00 (m, 1H), 8.08 (m, 1H); MS (FAB) 539 (M.sup.++1); Anal. calcd.
for C.sub.28H.sub.31FN.sub.4O.sub.6.2.75H.sub.2O: C, 57.18; H,
6.26; N, 9.53. Found, C, 57.20; H, 5.62; N, 9.06.
Example 227
3-dimethylamino-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylami-
no]ethoxy]benzoic acid
[1312] ##STR1900##
[1313] A mixture of pentafluorophenyl
[4-[N'-(2-methylphenyl)ureido]phenyl]acetate (1.42 g, 3.15 mmol),
methyl 4-[2-(N-methyl-2-amino)ethoxy]-3-nitro benzoate (800 mg,
3.15 mmol) and triethylamine (0.66 mL, 4.73 mmol) in DMF (8 mL) was
stirred at 50.degree. C. for 15 hr. The mixture was poured into
ice-1N HCl and extracted with CHCl.sub.3. The extract was brine,
dried over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with CHCl.sub.3:EtOAc (95:5 to 0:100,
v/v) as eluent to give 1.04 g (63%) methyl
3-nitro-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]etho-
xy]benzoate as a pale yellow oil. .sup.1H-NMR (CDCl.sub.3, 400 MHz)
.delta. 2.30 (s, 3H), 2.90, 3.02 and 3.05 (s, total 1H), 3.22 (s,
2H), 3.71 (s, 1H), 3.85 (3H), 3.93 (s, 3H), 4.19 and 4.39 (m, 2H),
7.02 (m, 1H), 7.19 (m, 4H), 7.35 (d, 1H, J=8.3 Hz), 7.40 (d, 1H,
J=8.0 Hz), 7.55 (s, 2H), 7.70 (m, 1H), 8.22 (d, 1H, J=6.7 Hz), 8.51
(s, 1H); MS (FAB) m/z 521 (M.sup.++1).
[1314] A mixture of methyl
3-nitro-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]etho-
xy]benzoate (1.04 g, 2 mmol) and 5%-Pd--C (1.2 g) in THF-MeOH--AcOH
(1:1:1, v/v, 150 mL) was hydrogenated at 45 psi for 18 hr.
Insoluble catalyst was removed with suction, and the filtrate was
evaporated in vacuo to afford methyl
3-amino-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]etho-
xy]benzoate as a pale yellow gum.
[1315] To a stirred solution of
3-amino-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]etho-
xy]benzoate, formaldehyde (5 mL), and AcOH (1.14 mL, 20 mmol) in
MeCN (5 mL) was added NaBH.sub.3CN (1.26 g, 20 mmol) at room temp,
and the resulting mixture was stirred for 15 hr at the same temp.
Sat. NaHCO.sub.3 was added to the mixture and extracted with
CHCl.sub.3. The extract was washed with brine, dried over
MgSO.sub.4, and evaporated in vacuo. The residue was
chromatographed on silica-gel with toluene:acetone (7:3 to 1:1,
v/v) as eluent to give 85 mg (2 steps, 8%) methyl
3-dimethylamino-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl
amino]ethoxy]benzoate as an oil. .sup.1H-NMR (CDCl.sub.3, 400 MHz)
.delta. 2.12 (s, 3H), 2.73 (s, 3H), 2.75 (s, 3H), 3.05 (s, 1H),
3.20 (s, 2H), 3.60 (s, 1H), 3.80 (m, 3H), 3.88 (s, 3H), 4.17 (r,
2H), 6.95-7.28 (m, 8H), 7.55-7.75 (m, 3H); MS (FAB) m/z 518
(M.sup.++1).
[1316] A stirred mixture of methyl
3-methylamino-4-[[2-[4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamin-
o]ethoxy]benzoate (ap315201) (85 mg, 0.16 mmol) in THF (15 mL) and
1N NaOH (0.32 mL, 0.32 mmol) was heated under reflux for 15 hr. The
pH of the mixture was adjusted to 5.0 by the addition of 1N HCl,
and extracted with CHCl.sub.3:MeOH (9:1, v/v). The extract was
washed with brine, dried over MgSO.sub.4, and evaporated in vacuo.
The residue was crystallized from Et.sub.2O to give 53 mg (65%) 278
as a white crystalline material. mp 110-115.degree. C.; .sup.1H-NMR
(CD.sub.3OD, 400 MHz) .delta. 2.29 (s, 3H), 2.75 (s, 3H), 2.76 (s,
3H), 3.02 (s, 1H), 3.20 (s, 2H), 3.72 (s, 1H), 3.85 (m, 3H), 4.18
and 4.28 (m, total 2H), 6.95-7.03 (m, 2H), 7.18 (m, 4H), 7.34 (d,
1H, J=8.3 Hz), 7.38 (d, 1H, J=8.8 Hz), 7.62 (d, 1H, J=8.3 Hz), 7.66
(s, 1H), 7.80 (m, 1H); MS (FAB) m/z 505 (M.sup.++1).
Example 228
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacety-
l]methylamino]ethoxy]benzoic acid
[1317] ##STR1901##
[1318] To a stirred cold (0.degree. C.) solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-fluoro
phenyl)ureido]phenylacetyl]methylamino]ethoxy]benzoate (100 mg,
0.19 mmol) in acetone/AcOH/DMF (13 mL, 6:6:1, v/v/v) was added
NaBH.sub.3CN (300 mg), and the resulting mixture was stirred for 60
hr at room temp. The mixture was pored into sat. NaHCO.sub.3 and
the solid was collected with suction. The precipitate was dissolved
in CHCl.sub.3 (20 mL), and the solution was washed with brine,
dried over MgSO.sub.4, and evaporated under a reduced pressure. The
residue was chromatographed on silica-gel plate with
toluene:acetone (2:1, v/v) as eluent to give 108 mg (100%) methyl
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phe-
nylacetyl]methylamino]ethoxy]benzoate as a colorless oil.
.sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 1.20 (m, 1H), 2.88 (s,
6H), 3.02 and 3.12 (s, total 3H), 3.53 (s, 2H), 3.60-3.80 (m, 7H),
3.85 (s, 3H), 4.10 and 4.20 (m, 2H), 6.65-675 (m, 2H), 6.90-7.08
(m, 2H), 7.22-7.35 (m, 2H), 8.02 (s, 2H), 8.10 (m, 2H), 8.21 (br,
1H), 8.33 (br, 1H); MS (FAB) m/z 566 (M.sup.++1).
[1319] A stirred mixture of methyl
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacet-
yl]methylamino]ethoxy]benzoate (116 mg, 0.2 mmol), 0.25 N NaOH (6
mL), and THF (6 mL) was heated under reflux for 8 hr. The mixture
was poured into water (200 mL), acidified by 1N HCl and the solid
was collected with suction. The solid was recrystallized from
CHCl.sub.3-n-hexane-diisopropylether to give 56 mg (50%) 279 as a
colorless crystalline powder. mp 195-200.degree. C.; .sup.1H-NMR
(CD.sub.3OD, 400 MHz) .delta. 1.15-1.20 (m, 6H), 3.01 (s, 1H), 3.12
(s, 2H), 3.48-3.60 (m, 1H), 3.68 (s, 3H), 3.75 (s, 2H), 3.82 (s,
1H), 3.86 (m, 3H), 4.15-4.23 (m, 2H), 6.80 (m, 3H), 4.15-4.23 (m,
2H), 6.80 (m, 3H), 6.98 (m, 1H), 7.10 (m, 2H), 7.20 and 7.25 (s,
total 1H), 7.32 (m, 1H), 7.98 (m, 1H), 8.05 (m, 1H); MS (FAB) m/z
552 (M+H).sup.+; Anal. calcd. for
C.sub.29H.sub.33FN.sub.4O.sub.6.1.0H.sub.2O: C, 61.04; H, 6.18; N,
9.82. Found, C, 61.36; H, 6.25; N, 9.45.
Example 229
4-[[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]-2-methylamino]-
-2-methyl-2-propoxy]benzoic acid
[1320] ##STR1902##
[1321] To a stirred solution of 2-amino-2-methyl-1-propanol (8.4 g,
93.89 mmol) and triethylamine (11.4 g, 0.113 mol) in DMF-water
(1:1, v/v, 100 mL) was added di-tert-butyl dicarbonate (25 g, 0.15
mol) at 5 to 10.degree. C. The resulting solution was stirred for 2
hr at room temp. The mixture was diluted with water (100 mL) and
extracted with EtOAc. The extract was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was chromatographed
on silica-gel with CH.sub.2Cl.sub.2 as eluent to give 12 g (68%)
2-tert-butoxycarbonylamino-2-methyl-1-propanol as a syrup.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.25 (s, 6H), 1.43 (s, 9H), 3.59
(d, J=8.3 Hz, 2H), 4.68 (br, 1H).
[1322] To a stirred suspension of
2-tert-butoxycarbonylamino-2-methyl-1-propanol (5.7 g, 30.11 mmol)
and powdered NaOH (6.7 g, 0.151 mol) in Et.sub.2O (200 mL) was
added p-toluenesulfonyl chloride (6.9 g, 36.14 mmol) at room temp.
The stirred resulting mixture was heated under reflux for 8 hr.
After cooling, ice-water (100 mL) was added to the solution.
Separated Et.sub.2O layer was washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. To the residue was added n-hexane
and triturated. The solid was collected to afford 8.5 g (82.2%)
2-tert-butoxycarbonylamino-2-methyl-1-propyl p-toluenesulfonate as
a crystalline material. .sup.1H-NMR (CDCl.sub.3) .delta. 1.26 (s,
6H), 1.38 (s, 9H), 2.37 (s, 3H), 4.05 (s, 2H), 4.49 (br, 1H), 7.34
(d, J=7.8 Hz, 2H), 7.78 (d, J=7.8 Hz, 2H).
[1323] A stirred mixture of
2-tert-butoxycarbonylamino-2-methyl-1-propyl p-toluenesulfonate
(8.2 g, 23.88 mmol) and powdered NaOH (6.7 g, 0.151 mol) in
Et.sub.2O (200 mL) was heated under reflux for 10 hr. After
cooling, the mixture was filtered. And the filtrate was washed with
water, brine, dried over Na.sub.2SO.sub.4, and evaporated. The
residue was chromatographed on silica-gel with n-hexane:EtOAc (6:1,
v/v) as eluent to give 2.7 g (66.2%)
1-tert-butoxycarbonyl-2-methylpropylene imine as an oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.29 (s, 6H), 1.47 (s, 9H), 2.05
(s, 2H).
[1324] To a stirred solution of
1-tert-butoxycarbonyl-2-methylpropyleneimine (1.03 g, 6.01 mmol)
and methyl 4-hydroxybenzoate (800 mg, 6.26 mmol) in
CH.sub.2Cl.sub.2 (10 mL) was added boron trifluoride diethyl ether
(0.127 mL, 1 mmol) at ambient temp. The resulting solution was
stirred for a further 3 hr at the same temp. The mixture was washed
with water, brine, dried over Na.sub.2SO.sub.4, and evaporated. The
residue was chromatographed on silica-gel with n-hexane:EtOAc (6:1,
v/v) as eluent to give 550 mg (33%) methyl
4-(1-tert-butoxycarbonylamino-2-methyl-2-propoxy)benzoate as a gum.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.33 (s, 6H), 1.47 (s, 9H), 3.36
(d, J=6.3 Hz, 2H), 3.90 (s, 3H), 5.05 (br, 1H), 6.99 (d, J=8.8 Hz,
2H), 7.97 (d, J=8.8 Hz, 2H).
[1325] A mixture of methyl
4-(1-tert-butoxycarbonylamino-2-methyl-2-propoxy)benzoate (460 mg,
1.42 mmol) and anisole (0.155 mL, 1.42 mmol) in CH.sub.2Cl.sub.2
(15 mL) and TFA (3 mL) was stirred for 3 hr at room temp. The
mixture was evaporated off. The residue was dissolved in
CH.sub.2Cl.sub.2 (30 mL) and made basic by the addition of 0.5N
NaOH, The CH.sub.2Cl.sub.2 layer was separated, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was chromatographed
on silica-gel with CH.sub.2Cl.sub.2 as eluent to give 370 mg (100%)
methyl 4-(1-amino-2-methyl-2-propoxy)benzoate as a gum. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.34 (s, 6H), 2.87 (s, 2H), 3.90 (s, 3H), 7.10
(d, J=8.8 Hz, 2H), 7.97 (d, J=8.8 Hz, 2H).
[1326] To a stirred solution of methyl
4-(1-amino-2-methyl-2-propoxy)benzoate (370 mg, 1.66 mmol) and
triethylamine (0.35 mL, 2.49 mmol) in CH.sub.2Cl.sub.2 (15 mL) was
added trifluoroacetic anhydride (0.316 mL, 2.24 mmol) at 0.degree.
C. After stirred for 1 hr at the same temp, water was added to the
solution. CH.sub.2Cl.sub.2 layer was separated, washed with water,
dried over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica-gel (20 mL) with CH.sub.2Cl.sub.2 as
eluent to give 530 mg (100%) methyl
4-(1-trifluoroacetamido-2-methyl-2-propoxy)benzoate as a gun. This
compound was used to the subsequent reaction without further
purification.
[1327] To a stirred mixture of methyl
4-(1-trifluoroacetamido-2-methyl-2-propoxy)benzoate (530 mg, 1.66
mmol) and K.sub.2CO.sub.3 (345 mg, 2.49 mmol) in DMF (10 mL) was
added MeI (0.14 mL, 2.37 mmol) at room temp. The resulting mixture
was stirred for 18 hr at room temp. The mixture was poured into
water, and extracted with EtOAc. The extract was washed with washed
with brine, dried over Na.sub.2SO.sub.4, and evaporated. The
residual gum was used to the subsequent reaction without further
purification.
[1328] The above crude residue was dissolved in MeOH (10 mL). To
the stirred solution was added water (5 mL) and Na.sub.2CO.sub.3
(352 mg, 3.32 mmol), and the resulting mixture was stirred for 5 hr
at room temp. The mixture was poured into water and extracted with
CHCl.sub.3. The extract was washed with water, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was chromatographed
on silica-gel with CHCl.sub.3 as eluent to give 390 mg (100%)
methyl 4-(1-methylamino-2-methyl-2-propoxy)benzoate as a gum.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.37 (s, 6H), 2.51 (s, 3H), 3.89
(s, 3H), 6.92 (d, J=8.8 Hz, 2H), 7.97 (d, J=8.8 Hz, 2H).
[1329] To a stirred mixture of methyl
4-(1-methylamino-2-methyl-2-propoxy)benzoate (200 mg, 0.84 mmol),
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetic acid (268 mg,
0.84 mmol), 4-DMAP (125 mg, 1.0 mmol) in DMF (5 mL) was added EDC
(220 mg, 1.14 mmol) at ambient temp. The resulting mixture was
stirred for a further 10 hr at ambient temp. The mixture was poured
into water, and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated. The residual
gum was triturated with CH.sub.2Cl.sub.2 and Et.sub.2O to give 200
mg (44.1%) methyl
4-[[1-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]methylamino]-2-
-methyl-2-propoxy]benzoate as a crystalline material. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.36 and 1.31 (each s, 6H), 3.24-3.88 (series
of s, 13H), 6.65-8.20 (series of m, 14H).
[1330] A mixture of methyl
4-[[1-[4-[1-(2-fluorophenyl)ureido]-3-methoxyphenylacetyl]methylamino]-2--
methyl-2-propoxy]benzoate (180 mg, 0.335 mmol) in THF (3 mL) and
0.25N NaOH (4 mL) was stirred for 5 hr at room temp. The mixture
was poured into ice-1N HCl (5 mL). The solid was collected, washed
with water, and air-dried. The crude solid was recrystallized from
EtOH--CHCl.sub.3-n-hexane to give 70 mg (40%) 280 as fine needles.
mp 200-207.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.26 and
1.33 (each s, 6H), 3.70-3.81 (series of s, 7H), 3.83 (s, 3H),
6.75-8.20 (series of m, 10H), 8.71 (s, 1H), 9.17 (br s, 1H), 12.72
(s, 1H).
Example 230
(S)-3-chloro-4-[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetylamin-
o]-1-propoxy]benzoic acid
[1331] ##STR1903##
[1332] A mixture of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (327 mg,
1.03 mmol), methyl (S)-3-chloro-4-(2-amino-1-propoxy)benzoate (250
mg, 1.03 mmol), EDC (hydrochloride) (295 mg, 1.54 mmol), HOBt (208
mg, 1.54 mmol), and DMAP (25 mg, 0.20 mmol) in DMF (8 mL) was
stirred at room temp overnight. The mixture was diluted with EtOAc,
washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4 and
evaporated. The residue was recrystallized from CHCl.sub.3-EtOAc to
give 308 mg (55%) as a white crystalline powder. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.29 (d, 3H, J=6.8 Hz), 3.51 (s, 2H), 3.84 (s,
3H), 3.88 (s, 3H), 4.01-4.08 (m, 2H), 4.38-4.39 (m, 1H), 6.25 (d,
1H, J=8.3 Hz), 6.78-6.83 (m, 2H), 6.90-6.95 (m, 2H), 7.02-7.11 (m,
2H), 7.89 (dd, 1H, J=2.0, 8.8 Hz), 8.02 (d, 1H, J=2.4 Hz), 8.20 (d,
1H, J=8.3 Hz), 8.27-8.31 (m, 1H), 8.53 (s, 1H), 8.84 (s, 1H).
[1333] To a stirred solution of methyl
(S)-3-chloro-4-[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetylami-
no]-1-propoxy]benzoate (308 mg, 0.57 mmol) in THF-MeOH (10 mL, 1:1,
v/v) was added 0.5 N NaOH (10 mL), and the reaction mixture was
heated under reflux for 1 hr. The mixture was poured into ice-1 N
HCl and the solid was collected. The crude solid was purified by
recrystallization from MeOH--CHCl.sub.3-n-hexane to give 196 mg
(65%) 281 as a white crystalline powder. mp 188-191.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.21 (d, 3H, J=6.4 Hz), 3.38 (s,
2H), 3.83 (s, 3H), 4.02-4.18 (m, 3H), 6.78-7.29 (series of m, total
6H), 7.85-8.00 (m, 3H), 8.12-8.19 (m, 2H), 8.70 (s, 1H), 9.17 (s,
1H), 12.98 (bs, 1H); MS (FAB) m/z 530 (M.sup.+), 531 (M.sup.++1),
532 (M.sup.++2); Anal. Calcd for
C.sub.26H.sub.25ClFN.sub.3O.sub.6.1/4H.sub.2O: C, 58.43; H, 4.81;
Cl, 6.63; F, 3.55; N, 7.86. Found: C, 58.45; H, 4.83; Cl, 6.68; F,
3.38; N, 7.79.
Example 231
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]eth-
ylamino]benzoic acid
[1334] ##STR1904##
[1335] A stirred solution of
N-benzyloxycarbonyl-N-methylaminoacetaldehyde (1.77 g, 1.1 mmol) in
toluene (3 mL) was added methyl-4-aminobenzoate (1.29 g, 8.5 mmol),
and the mixture was stirred for 1 hr at room temp. The reaction
mixture was evaporated under a reduced pressure and the residue was
dissolved into MeCN (15 mL). To the solution was added AcOH (2.44
mL, 43 mmol) and NaBH.sub.3CN (2.67 g, 43 mmol), and the resulting
mixture was stirred for 15 hr at room temp. The reaction was
quenched by the addition of sat. NaHCO.sub.3. The mixture was
extracted with EtOAc, washed with brine, dried over MgSO.sub.4, and
evaporated. The residue was chromatographed on silica-gel with
n-hexane:EtOAc (7:3, v/v) as eluent to give 1.26 g (43%) methyl
4-[2-(N-benzyloxy carbonyl-N-methylamino)-ethylamino]benzoate as a
colorless oil. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.96 (s,
3H), 3.32 (br m, 2H), 3.50-3.60 (m, 2H), 3.82 (s, 3H), 5.15 (each
d, 2H, J=14.6 Hz), 6.35 and 6.58 (m, total 2H), 7.36 (s, 5H), 7.76
and 7.84 (m, total 2H); MS (FAB) m/z 342 (M.sup.++1).
[1336] To a stirred solution of methyl
4-[2-(N-benzyloxycarbonyl-N-methylamino)ethylamino]benzoate (1.26
g, 3.68 mmol) in MeOH (20 mL) was added 5 wt. Pd --C (700 mg), and
the mixture was hydrogenated (3 atm) for 4 hr at room temp. The
mixture was filtered, and the filtrate was evaporated under a
reduced pressure to give 600 mg (78%) methyl
4-[2-(N-methylamino)ethylamino]benzoate as a colorless oil.
.sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.46 (s, 3H), 2.88 (t,
2H, J=5.5 Hz), 3.27 (br s, 2H), 3.85 (s, 3H), 6.57 (d, 2H, J=8.8
Hz), 7.85 (d, 2H, J=8.8 Hz); MS (FAB) m/z 209 (M.sup.++1).
[1337] To a stirred solution of methyl
4-[2-(N-methylamino)ethylamino]benzoate (590 mg, 2.83 mmol)
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (891 mg,
2.83 mmol) in DMF (14 mL) was added EDC (815 mg, 4.25 mmol), HOBt
(574 mg, 4.25 mmol), and 4-DMAP (519 mg, 4.25 mmol), and the
resulting mixture was stirred overnight at room temp. The mixture
was poured into 1N HCl and the solid was collected with suction.
The crude solid was purified by chromatography on silica-gel
(middle pressure) with CHCl.sub.3-EtOAc (10:0 to 7:3, v/v) as
eluent to give 1.25 g (87%) methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]et-
hylamino]benzoate as a light yellow oil. .sup.1H-NMR (CDCl.sub.3,
400 MHz) .delta. 3.18 (s, 2H), 3.05 and 3.32 (s, total 2H),
3.72-3.85 (m, 9H), 4.12 and 4.23 (m, total 2H), 6.78 (m, 3H), 7.05
(t, 1H, J=7.5 Hz), 7.20 (m, 2H), 7.43-7.50 (m, 3H), 7.62 (d, 1H,
J=8.8 Hz), 8.05 (d, 1H, J=8.8 Hz); MS (FAB) m/z 505
(M.sup.++1).
[1338] A stirred mixture of methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]et-
hylamino]benzoate (300 mg, 0.6 mmol) in 0.25 N NaOH (6 mL) and THF
(6 mL) was heated under reflux for 8 hr. The mixture was poured
into water, and acidified with 1N HCl. The solid was collected with
suction. The crude solid was recrystallized from
n-hexane-diisopropylether to give 202 mg (69%) 282 as a pale yellow
crystalline powder. mp 115-120.degree. C.; IR (KBr) 3346, 2935,
1603, 1531, 1454, 1417, 1257, 1174, 1036, 754 cm.sup.-1;
.sup.1H-NMR (CD.sub.3OD, 400 MHz) .delta. 2.28 (s, 3H), 2.98 (s,
1H), 3.10 (s, 2H), 3.35-3.42 (m, 2H), 3.53-3.65 (m, 3H), 3.70 (s,
1H), 3.80 and 3.82 (s, 3H), 6.60-6.85 (m, 4H), 7.02 (m, 1H), 7.18
(m, 2H), 7.58 (m, 1H), 7.82 (m, 2H), 7.96 (m, 1H); MS (FAB) m/z 491
(M.sup.++1).
Example 232
(S)-3-chloro-4-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phen-
ylacetyl]amino]-1-propoxy]benzoic acid
[1339] ##STR1905##
[1340] To a cooled (0.degree. C.) solution of methyl
(S)-3-chloro-4-(2-amino-1-propoxy)benzoate (1.74 g, 7.14 mmol) in
CH.sub.2Cl.sub.2 (20 mL) was added Et.sub.3N (1.19 mL, 8.54 mmol)
and trifluoroacetic anhydride (TFAA) (1.11 mL, 7.86 mmol), and the
reaction mixture was stirred at room temp overnight. The mixture
was diluted with CHCl.sub.3, washed with 0.5 N HCl, brine, dried
over Na.sub.2SO.sub.4 and evaporated. The residue was
recrystallized from CHCl.sub.3-n-hexane to give 1.59 g (66%)
(S)-3-chloro-4-(2-trifluoroacetamido-1-propoxy)benzoate as a white
crystalline material. mp 122-125.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.48 (d, 3H, J=6.8 Hz), 3.91 (s, 3H),
4.10-4.19 (m, 2H), 4.47-4.52 (m, 1H), 6.68 (bs, 1H), 6.92-6.94 (m,
1H), 7.93-7.95 (m, 1H), 8.07-8.08 (m, 1H); MS (FAB) m/z 340
(M.sup.++1); Anal. Calcd for C.sub.13H.sub.13ClF.sub.3NO.sub.4: C,
45.96; H, 3.86; Cl, 10.44; F, 16.78; N, 4.12. Found: C, 45.88; H,
3.97; Cl, 10.24; F, 16.72; N, 4.18.
[1341] To a stirred solution of methyl
(S)-3-chloro-4-(2-trifluoroacetamido-1-propoxy)benzoate (800 mg,
2.36 mmol) in DMF (5 mL) was added K.sub.2CO.sub.3 (651 mmol, 4.71
mmol) and MeI (0.22 mL, 3.53 mmol), and the reaction mixture was
stirred at 60.degree. C. overnight. The mixture was diluted with
EtOAc, washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4
and evaporated. The residue was purified by column chromatography
on silica-gel with 5% EtOAc in CHCl.sub.3 as eluent to give 850 mg
(100%) methyl
(S)-3-chloro-4-[2-(N-methyl-N-trifluoroacetamido)-1-propoxy]benzoate
as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.43-1.46 (m,
3H), 3.03 and 3.21 (s, 3H), 3.90 (s, 3H), 4.04-4.22 (m, 2H),
4.81-4.87 (m, 1H), 6.91 (d, 1H, J=8.3 Hz), 7.93 (dd, 1H, J=2.0,
8.3H), 8.06 (d, 1H, J=2.0 Hz).
[1342] To a stirred solution of methyl
(S)-3-chloro-4-[2-(N-methyl-N-trifluoroacetamido)-1-propoxy]benzoate
(880 mg, 2.49 mmol) in MeOH--H.sub.2O (10 mL, 1:1, v/v) was added
K.sub.2CO.sub.3 (516 mg, 3.73 mmol), and the resulting mixture was
stirred at room temp overnight. The mixture was diluted with EtOAc,
washed with H.sub.2O, brine, dried over Na.sub.2SO.sub.4 and
evaporated to give 460 mg (72%)
(S)-3-chloro-4-(2-methylamino-1-propoxy)benzoate as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.20 (d, 3H, J=6.4 Hz), 2.51
(s, 3H), 3.07-3.12 (m, 1H), 3.89 (s, 3H), 3.92-4.04 (m, 2H),
6.93-6.95 (m, 1H), 7.90-7.93 (m, 1H), 8.05-8.06 (m, 1H).
[1343] A mixture of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (296 mg,
0.93 mmol), methyl (S)-3-chloro-4-(2-methylamino-1-propoxy)benzoate
(240 mg, 0.93 mmol), EDC (hydrochloride) (268 mg, 1.40 mmol), HOBt
(189 mg, 1.40 mmol), and DMAP (23 mg, 0.19 mmol) in DMF (8 mL) was
stirred at room temp for 1.5 hr. The mixture was diluted with
EtOAc, washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4
and evaporated. The residue was purified by column chromatography
on silica-gel with 5% MeOH in CHCl.sub.3 as eluent to give 520 mg
(100%) methyl
(S)-3-chloro-4-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phe-
nylacetyl]amino]-1-propoxy]benzoate as a red-brown amorphous
solid.
[1344] To a stirred solution of this product (520 mg, 0.93 mmol) in
THF (10 mL) was added 0.5 N NaOH (10 mL), and the resulting mixture
was heated under reflux for 3 hr. The mixture was poured into ice-1
N HCl and the solid was collected. The crude solid was
recrystallized from CHCl.sub.3-Et.sub.2O to give 170 mg (2 steps,
37%) 283 as a white crystalline powder. mp 142-147.degree. C.;
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.13-1.20 (m, 3H), 2.74 and 2.94
(s, 3H), 3.65 (s, 2H), 3.82 and 3.84 (s, 3H), 4.13-4.22 (m, 2H),
4.53 and 4.91-4.92 (m, 1H), 6.71-7.29 (series of m, total 6H),
7.86-8.02 (m, 3H), 8.15-8.19 (m, 1H), 8.71 (s, 1H), 9.17 (s, 1H),
13.00 (bs, 1H); MS (FAB) m/z 544 (M.sup.+), 545 (M.sup.++1); Anal.
Calcd for C.sub.27H.sub.27ClFN.sub.3O.sub.6.1/4H.sub.2O: C, 59.13;
H, 5.05; Cl, 6.46; F, 3.46; N, 7.66. Found: C, 59.19; H, 4.99; Cl,
6.64; F, 3.23; N, 7.55.
Example 233
(S)-3-chloro-4-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phen-
ylacetyl]amino-1-propoxy]benzoic acid
[1345] ##STR1906##
[1346] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (261 mg,
0.83 mmol), methyl (S)-3-chloro-4-(2-methylamino-1-propoxy)benzoate
(214 mg, 0.83 mmol), EDC (hydrochloride) (239 mg, 1.25 mmol), HOBt
(168 mg, 1.24 mmol), and DMAP (20 mg, 0.16 mmol) in DMF (8 mL) was
stirred at room temp for 2 hr. The mixture was diluted with EtOAc,
washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4 and
evaporated. The residue was purified by column chromatography on
silica-gel with CHCl.sub.3-MeOH (50:1, v/v) as eluent to give 470
mg (100%) methyl
(S)-3-chloro-4-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phe-
nylacetyl]amino]-1-propoxy]benzoate as a pale yellow amorphous
solid.
[1347] To a stirred solution of the above product (470 mg, 0.95
mmol) in THF (10 mL) was added 0.5 N NaOH (10 mL) and the reaction
mixture was heated under reflux for 3 hr. The mixture was poured
into ice-1 N HCl and the solid was collected. The crude solid was
recrystallized from CHCl.sub.3-Et.sub.2O to give 168 mg (2 steps,
33%) 284 as a pale yellow crystalline powder. mp 125-130.degree.
C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.13-1.19 (m, 3H), 2.24 (s,
3H), 2.74 and 2.94 (s, 3H), 3.65 (s, 2H), 3.83 and 3.85 (s, 3H),
4.14-4.22 (m, 2H), 4.91-4.93 (m, 1H), 6.70-6.74 (m, 1H), 6.84 (m,
1H), 6.92-6.95 (m, 1H), 7.11-7.17 (m, 2H), 7.25-7.29 (m, 1H),
7.78-7.80 (m, 1H), 7.85-7.93 (m, 2H), 7.99-8.02 (m, 1H), 8.46 (s,
1H), 8.56 (s, 1H), 12.99 (bs, 1H); MS (FAB) m/z 540 (M.sup.+), 541
(M.sup.++1); Anal. Calcd for
C.sub.28H.sub.30ClN.sub.3O.sub.6.1/2H.sub.2O: C, 61.26; H, 5.69; N,
7.65. Found: C, 61.15; H, 5.58; N, 7.51.
Example 234
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacety-
l]methylamino]ethoxy]benzoic acid
[1348] ##STR1907##
[1349] To a cold (0.degree. C.), stirred solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]methylamino]ethoxy]benzoate (300 mg,
0.58 mmol) in acetone-AcOH-DMF (13 mL, 6:6:1, v/v/v) was added
NaBH.sub.3CN (300 mg) and the resulting mixture was stirred for 60
hr at room temp. The mixture was poured into sat. NaHCO.sub.3 and
extracted with CHCl.sub.3. The extract was washed with brine, dried
over MgSO.sub.4, and evaporated to give 280 mg (86%) methyl
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]methylamino]ethoxy]benzoate as a colorless oil. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 1.21 (m, 6H), 2.30 (s, 3H), 3.05 and
3.10 (s, total 2H), 3.59 (s, 3H), 3.62-3.81 (m, 6H), 3.89 (s, 3H),
4.10 and 4.21 (m, 2H), 6.65-6.80 (m, 4H), 7.10 (m, 1H), 7.20 (s,
3H), 7.32 (m, 2H), 7.54 (d, 1H, J=8.3 Hz), 8.00 (s, 1H), 8.07 (d,
1H, J=8.3 Hz); MS (FAB) m/z 563 (M.sup.++1).
[1350] A stirred mixture of methyl
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]methylamino]ethoxy]benzoate (280 mg, 0.5 mmol), 0.25 N NaOH (6
mL), and THF (6 mL) was heated under reflux for 8 hr. The mixture
was poured into water (200 mL), acidified with 1N HCl and the solid
was collected with suction. The solid was recrystallized from
CHCl.sub.3-n-hexane-diisopropylether to give 202 mg (74%) 285 as a
light yellow crystalline powder. mp 130-135.degree. C.; .sup.1H-NMR
(CD.sub.3OD, 400 MHz) .delta. 1.18 and 1.22 (d, total 6H, J=6.3
Hz), 2.29 and 2.32 (s, total 3H), 3.04 and 3.14 (s, total 2H),
3.60-3.90 (m, 9H), 4.16 and 4.25 (m, total 2H), 6.80 (m, 3H), 7.02
(m, 1H), 7.12-7.24 (m, 3H), 7.29-7.38 (m, 1H), 7.59 (m, 1H), 8.02
(m, 1H); MS (FAB) m/z 548 (M+H).sup.+; Anal. calcd. for
C.sub.30H.sub.36N.sub.4O.sub.6: C, 65.68; H, 6.61. Found: C, 65.80;
H, 6.83.
Example 235
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]eth-
yl]methylamino benzoic acid
[1351] ##STR1908##
[1352] To a stirred cold (0.degree. C.) solution of methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methylamino]et-
hyl]aminobenzoate (300 mg, 0.6 mmol) in MeCN-formaldehyde-AcOH (11
mL, 8:2:1, v/v/v) was added NaBH.sub.3CN (187 mg, 2.40 mmol), and
the resulting mixture was stirred for 18 hr at room temp. The
mixture was poured into sat. NaHCO.sub.3 and extracted with
CHCl.sub.3. The extract was washed with brine, dried over
MgSO.sub.4, and evaporated to give 309 mg (100%) methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-N'-methylam-
ino]ethyl]N-methylaminobenzoate as a colorless oil. .sup.1H-NMR
(CDCl.sub.3, 400 MHz) .delta. 2.30 (m, 3H), 2.98 (m, 7H), 3.46-3.72
(m, 9H), 3.82 (m, 3H), 6.32 (m, 1H), 6.55-6.75 (m, 4H), 7.05-7.50
(m, 2H), 7.82-8.02 (m, 4H); MS (FAB) m/z 518 (M.sup.++1).
[1353] A stirred mixture of methyl
4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-2-N'-methylam-
ino]ethyl]methylaminobenzoate (309 mg, 0.6 mmol) in 1 N NaOH (2.4
mL), and THF (10 mL) was heated under reflux for 8 hr. The mixture
was poured into water (200 mL), acidified by the addition of 1N
HCl. The solid was collected with suction. The crude solid was
recrystallized from CHCl.sub.3-n-hexane-diisopropylether to give
211 mg (70%) 286 as a light yellow crystalline powder. mp
125-130.degree. C.; IR (KBr) 3338, 2933, 1601, 1529, 1182, 1036,
752 cm.sup.-1; .sup.1H-NMR (CD.sub.3OD, 400 MHz) .delta. 2.28 and
2.29 (s, 3H), 2.95-3.02 (m, 6H), 3.60 (br, 6H), 3.80 (s, 1H), 3.86
(s, 2H), 6.60-6.82 (m, 4H), 7.01-7.18 (m, 3H), 7.58 (m, 1H),
7.82-7.99 (m, 3H); MS (FAB) m/z 504 (M.sup.++1).
Example 236
(S)-3-chloro-4-[2-[N-benzyl-N-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phen-
ylacetyl]amino]-1-propoxy]benzoic acid
[1354] ##STR1909##
[1355] To a stirred solution of NaBH.sub.3CN (985 mg, 15.68 mmol)
in MeOH (5 mL) was added a solution of methyl
(S)-3-chloro-4-(2-amino-1-propoxy)benzoate (382 mg, 1.57 mmol) and
benzaldehyde (0.19 mL) in MeOH (5 mL), and the resulting mixture
was stirred at room temp overnight. The mixture was quenched by
H.sub.2O and extracted with CHCl.sub.3. The extract was washed with
brine, dried over Na.sub.2SO.sub.4 and evaporated. The residue was
purified by column chromatography on silica-gel with
CHCl.sub.3:MeOH (30:1, v/v) as eluent to give 336 mg (64%)
(S)-3-chloro-4-(2-N-benzylamino-1-propoxy)benzoate as a colorless
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.22 (d, 3H, J=6.4 Hz),
3.21-3.25 (m, 1H), 3.84-4.03 (m, total 7H), 6.89-6.91 (m, 1H),
7.23-7.37 (m, 5H), 7.89-7.91 (m, 1H), 8.05 (m, 1H).
[1356] A mixture of
3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetic acid (320 mg,
1.01 mmol), methyl
(S)-3-chloro-4-(2-N-benzylamino-1-propoxy)benzoate (336 mg, 1.01
mmol), EDC (hydrochloride) (289 mg, 1.51 mmol), HOBt (204 mg, 1.51
mmol) and DMAP (25 mg, 0.20 mmol) in DMF (7 mL) was stirred at room
temp for 2 days. The mixture was diluted with EtOAc, washed with
0.5 N HCl, brine, dried over Na.sub.2SO.sub.4 and evaporated. The
residue was purified by column chromatography on silica-gel with
CHCl.sub.3:MeOH (50:1, v/v) as eluent to give 607 mg (95%) methyl
(S)-3-chloro-4-[2-[N-benzyl-N-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phe-
nyl acetyl]amino]-1-propoxy]benzoate as a pale yellow amorphous
solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.20-1.27 (m, 3H), 3.62 (s,
2H), 3.73-4.16 (series of m, total 9H), 4.71 (bs, 2H), 6.67-7.38
(series of m, total 12H), 7.77-8.17 (series of m, total 5H).
[1357] To a stirred solution of methyl
(S)-3-chloro-4-[2-[N-benzyl-N-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phe-
nylacetyl]amino]-1-propoxy]benzoate (607 mg, 0.96 mmol) in THF (6
mL) was added 0.5 N NaOH (6 mL), and the resulting mixture was
heated under reflux overnight The mixture was poured into ice-1 N
HCl and the solid was collected. The crude solid was recrystallized
to give 192 mg (32%) 287 as a white crystalline powder. mp
125-130.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.07-1.23
(m, 3H), 3.76 (s, 2H), 3.85 (s, 3H), 3.90-4.26 (m, 3H), 4.56 (s,
2H), 6.66-7.38 (series of m, total 10H), 7.82-8.20 (series of m,
total 5H), 8.70-8.74 (m, 1H), 9.18-9.20 (m, 1H), 13.02 (bs, 1H); MS
(FAB) m/z 621 (M.sup.+30 1); Anal. Calcd for
C.sub.33H.sub.31ClFN.sub.3O.sub.6.1/4H.sub.2O: C, 63.46; H, 5.08;
Cl, 5.68; F, 3.04; N, 6.73. Found: C, 63.67; H, 5.16; Cl, 5.75; F,
2.95; N, 6.55.
Example 237
3-(N-isopropyl-N-methylamino)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureid-
o]phenylacetyl]methylamino]ethoxy]benzoic acid
[1358] ##STR1910##
[1359] To a stirred cold (0.degree. C.) solution of methyl
3-isopropylamino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]methylamino]ethoxy]benzoate (324 mg, 0.58 mmol) in
CH.sub.3CN-formaldehyde-AcOH (11 mL, 8:2:1, v/v/v) was added
NaBH.sub.3CN (145 mg, 2.30 mmol), and the resulting mixture was
stirred for 18 hr at room temp. The mixture was poured sat.
NaHCO.sub.3 and the solid was collected with suction. The crude
solid was dissolved in CHCl.sub.3 (20 mL), and the solution was
washed with brine, dried over MgSO.sub.4, and evaporated to give
317 mg (95%) methyl
3-(N-isopropyl-N-methylamino)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)urei-
do]phenylacetyl]methylamino]ethoxy]benzoate as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.02 (m, 6H), 2.29 (s, 3H), 2.63
(m, 3H), 3.02-3.20 (m, 3H), 3.49-3.80 (m, 8H), 3.88 (s, 3H), 4.06
and 4.21 (m, total 2H), 6.59 (m, 1H), 6.76 (m, 2H), 7.11-7.23 (m,
3H), 7.50-7.62 (m, 3H), 8.05 (d, 1H, J=8.3 Hz); MS (FAB) m/z 576
(M.sup.++1).
[1360] A stirred mixture of methyl
3-(N-isopropyl-N-methylamino)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)urei-
do]phenylacetyl]methylamino]ethoxy]benzoate (317 mg, 0.55 mmol) in
0.25 N NaOH (8.2 mL) and THF (8 mL) was heated under reflux for 8
hr. The mixture was poured into water (200 mL), acidified by the
addition of 1N HCl and the solid was collected with suction. The
crude solid was recrystallized from
CHCl.sub.3-n-hexane-diisopropylether to give 288 as a light yellow
crystalline powder. mp 130-135.degree. C.; IR (KBr) 2970, 1537,
1038, 754 cm.sup.-1; .sup.1H-NMR (CD.sub.3OD) .delta. 1.12 (m, 6H),
2.29 (s, 3H), 2.76 (m, 1H), 2.89 (s, 2H), 3.02 (s, 1H), 3.21 (s,
2H), 3.72 (s, 2H), 3.80 (s, 3H), 3.84 (m, 3H), 4.13 and 4.40 (m,
total 2H), 6.76 (d, 1H, J=7.8 Hz), 6.86 (s, 1H), 7.00 (m, 2H), 7.18
(m, 3H), 7.57 (d, 1H, J=7.8 Hz), 7.95 (m, 2H); MS (FAB) m/z 562
(M.sup.++1); Anal. Calcd for
C.sub.31H.sub.38N.sub.4O.sub.6.2.0H.sub.2O: C, 62.19; H, 7.07; N,
9.36. Found: C, 62.54; H, 6.85; N, 8.90.
Example 238
3-(1-piperidinyl)-4-[[2-[3-methoxy-4-[NP-(2-fluorophenyl)ureido]phenylacet-
yl]methylamino]ethoxy]benzoic acid
[1361] ##STR1911##
[1362] To a stirred solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (300 mg, 0.57 mmol) in THF (2 mL) was added a
solution of glutaraldehyde (50% aqueous solution) (0.13 mL, 0.69
mmol) in MeOH (1.4 mL), THF (0.993 mL), and 3N H.sub.2SO.sub.4
(0.952 mL) at 0.degree. C. To the solution was added NaBH.sub.3CN
(150 mg), DMF (5 mL), and MeOH (2 mL) at room temp, and the
resulting mixture was stirred for 15 hr. The mixture was poured
into sat. NaHCO.sub.3 and extracted with CHCl.sub.3. The extract
was washed with brine, dried over MgSO.sub.4, and evaporated under
a reduced pressure. The residue was chromatographed on silica gel
with toluene:acetone (7:3, v/v) as eluent to give 145 mg (43%)
methyl
3-(1-piperidinyl)-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]ph-
enylacetyl]methyl amino]ethoxy]benzoate as a colorless oil.
.sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 1.58-1.72 (m, 6H),
2.80-2.92 (m, 4H), 3.26 (s, 2H), 3.58 (s, 2H), 3.69 (s, 2H),
3.80-3.89 (m, 7H), 4.22 (m, 2H), 6.72 (s, 1H), 6.78 (m, 2H),
6.95-7.18 (m, 2H), 7.32 (s, 1 .mu.l), 7.60 (s, 1H), 7.63 (d, 1H,
J=7.8 Hz), 7.98 (d, 1H, J=7.8 Hz), 8.18 (m, 1H); MS (FAB) m/z 593
(M.sup.++1).
[1363] A stirred mixture of methyl
3-(1-piperidinyl)-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylace-
tyl]methylamino]ethoxy]benzoate (290 mg, 0.49 mmol), 1N NaOH (1.95
mL) in MeOH (5 mL) and THF (10 mL) was heated under reflux for 4
hr. The mixture was poured into water (200 mL), acidified with 1N
HCl (pH=4.0), and extracted with CHCl.sub.3-MeOH (9:1, v/v). The
combined extract was dried over MgSO.sub.4, and evaporated. The
residue was crystallized from diisopropylether-hexane to give 201
mg (71%) 289 as a light yellow crystalline powder. mp
125-130.degree. C.; IR (KBr) 3338, 2935, 1599, 1537, 1041, 752
cm.sup.-1; .sup.1H-NMR (CD.sub.3OD) .delta. 1.52-1.73 (m, 6H), 2.88
(s, 3H), 2.98 (br, 1H), 3.09 (s, 1H), 3.23 (s, 2H), 3.66 (s, 2H),
3.75 (s, 1H), 3.82 (s, 4H), 4.13 and 4.29 (m, total 2H), 6.78 (m,
2H), 6.99 (m, 2H), 7.10 (m, 2H), 7.60-7.70 (m, 2H), 7.96-8.07 (m,
2H); MS (FAB) m/z 578 (M.sup.+30 1); Anal. Calcd for
C.sub.31H.sub.35FN.sub.4O.sub.6.0.5H.sub.2O: C, 63.36; H, 6.17; N,
9.53. Found: C, 63.22; H, 6.15; N, 9.16.
Example 239
3-amino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyla-
mino]ethoxy]benzoic acid
[1364] ##STR1912##
[1365] To a stirred solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (300 mg, 0.58 mmol) in MeOH-THF (2:1, v/v, 12
mL) was added 0.25 N NaOH (9 mL), and the resulting mixture was
heated under reflux for 16 hr. The mixture was poured into water
(100 mL) and acidified by the addition of 1N HCl. The solid was
collected with suction. The residue was recrystallized from
diisopropylether to give 243 mg (83%) 290 as a yellow crystalline
powder. mp 125-130.degree. C.; IR (KBr) 3346, 2935, 1533, 1211,
1034, 756, 637 cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.29
(s, 3H), 3.05 (s, 1H), 3.72-3.85 (m, 7H), 4.12 and 4.25 (m, total
2H), 6.76-6.89 (m, 3H), 7.00 (m, 1H), 7.18 (m, 2H), 7.39 (m, 2H),
7.60 (d, 1H, J=7.8 Hz), 7.96 (m, 1H); Anal. Calcd for
C.sub.27H.sub.30N.sub.4O.sub.6.0.5H.sub.2O: C, 62.90; H, 6.06; N,
10.87. Found: C, 63.03; H, 6.14; N, 10.56.
Example 240
3-(1-piperidinyl)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacet-
yl]methylamino]ethoxy]benzoic acid
[1366] ##STR1913##
[1367] To a cooled and stirred solution of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (573 mg, 1.1 mmol) in THF-MeOH (2:1, v/v, 6
mL) was added glutaraldehyde (0.423 mL, 2.2 mmol), 3N
H.sub.2SO.sub.4 (1.83 mL, 5.5 mmol), and NaBH.sub.3CN (276 mg, 4.4
mmol). The resulting mixture was stirred for 18 hr at room temp.
The mixture was poured into sat. NaHCO.sub.3 (100 mL) and the solid
was collected with suction. The crude solid was purified by column
chromatography on silica-gel with toluene-acetone (10:0 to 4:1,
v/v) to give 240 mg (37%) methyl
3-(1-piperidinyl)-4-[[2-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetyl]methylamino]ethoxy]benzoate as a gum.
.sup.1H-NMR (CD.sub.3OD) .delta. 1.52-1.72 (m, 6H), 2.30 (s, 3H),
2.36 (s, 2H), 2.89 (br s, 3H), 2.98 (m, 1H), 3.05 (s, 1H), 3.20 (s,
2H), 3.68 (s, 2H), 3.69 (m, 2H), 3.79 (m, 2H), 3.88 (s, 2H), 4.08
and 4.21 (m, 2H), 6.35 and 6.42 (s, total 1H), 6.70 (s, 1H), 6.70
(s, 1H), 6.79 (m, 2H), 7.09-7.24 (m, 4H), 7.50-7.65 (m, 3H), 8.05
(d, 1H, J=8.3 Hz); MS (FAB) m/z 588 (M.sup.++1).
[1368] A stirred mixture of methyl
3-(1-piperidinyl)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylace-
tyl]methylamino]ethoxy]benzoate (240 mg, 0.41 mmol), 1 N NaOH (1.63
mL) in MeOH (6.5 mL), H.sub.2O (5 mL), and THF (6.5 mL) was heated
under reflux for 11 hr. The mixture was poured into water (200 mL),
acidified by the addition of 1N HCl until pH=4.0, and the solid was
collected with suction. The aqueous layer was extracted with
CHCl.sub.3-MeOH (9:1, v/v, 30 mL.times.3). The precipitate was
dissolved in the combined organic extract. The organic layer was
dried over MgSO.sub.4 and evaporated. The residue was crystallized
from diisopropylether to give 151 mg (65%) 291 as a light yellow
crystalline powder. mp 120-125.degree. C.; IR (KBr) 3354, 2935,
1535, 1252, 1217, 1034, 754, 638 cm.sup.-1; .sup.1H-NMR
(CD.sub.3OD) .delta. 1.55-1.72 (m, 6H), 2.30 (s, 3H), 2.89 (br,
2H), 2.98 (br, 1H), 3.09 (s, 1H), 3.22 (s, 2H), 3.69 (s, 3H), 3.74
(s, 1H), 3.83 (m, 4H), 4.12 and 4.28 (m, total 2H), 6.75 (m, 2H),
6.88-7.02 (m, 1H), 7.17 (m, 2H), 7.58-7.73 (m, 4H), 7.99 (d, 1H,
J=8.3 Hz); MS (FAB) m/z 574 (M.sup.++1); Anal. Calcd for
C.sub.32H.sub.38N.sub.4O.sub.6.0.5H.sub.2O: C, 65.85; H, 6.73; N,
9.60. Found: C, 65.94; H, 6.88; N, 9.03.
Example 241
3-amino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyla-
mino]ethoxy]benzoic acid
[1369] ##STR1914##
[1370] A stirred mixture of methyl
3-amino-4-[[2-[3-methoxy-4-[N'-(2-fluorophenyl)ureido]phenylacetyl]methyl-
amino]ethoxy]benzoate (250 mg, 0.48 mmol) in 1N NaOH (1.91 mL),
MeOH (8 mL), H.sub.2O (6 mL), and THF (8 mL) was heated under
reflux for 10 hr. The mixture was poured into water (200 mL),
acidified by the addition of 1N HCl (pH=4.8), and the solid was
collected with suction. The crude solid was recrystallized from
diisopropylether to give 209 mg (86%) 292 as a pale yellow
crystalline powder. mp 125-130.degree. C.; IR (KBr) 3325, 2935,
1537, 1209, 1032, 752, 449 cm.sup.-1; .sup.1H-NMR (CD.sub.3OD)
.delta. 3.05 (s, 1H), 3.31 (s, 2H), 3.77 (m, 3H), 3.80 (s, 1H),
3.84 (m, 3H), 4.14-4.26 (m, 2H), 6.80-6.87 (m, 3H), 7.01 (m, 1H),
7.10 (m, 2H), 7.39 (m, 2H), 7.80 (m, 1H), 8.07 (m, 1H); MS (FAB)
m/z 510 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.27FN.sub.4O.sub.6.0.5H.sub.2O: C, 60.11; H, 5.43; F,
3.66; N, 10.78. Found: C, 60.29; H, 5.40; F, 3.60; N, 10.59.
Example 242
(S)-3-amino-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamino-
]-1-propoxy]benzoic acid
[1371] ##STR1915##
[1372] To a stirred solution of
(S)-2-(N-tert-butoxycarbonylamino)-1-propanol (0.90 g, 5.13 mmol),
methyl 4-hydroxy-3-nitrobenzoate (1.01 g, 5.12 mmol), and Ph.sub.3P
(1.75 g, 6.67 mmol) in THF (15 mL) was added diisopropyl
azodicarboxylate (DIAD) (1.31 mL, 6.65 mmol) at room temp, and the
resulting mixture was heated under reflux overnight. The solution
was evaporated and the residue was dissolved in CH.sub.2Cl.sub.2
(20 mL) and TFA (10 mL). The mixture was stirred at room temp for 3
hr. The mixture was concentrated in vacuo. The residue was
dissolved in CHCl.sub.3, washed with sat. NaHCO.sub.3, brine, dried
over Na.sub.2SO.sub.4 and evaporated. The residue was purified by
column chromatography on silica-gel with 5% MeOH in CHCl.sub.3 as
eluent to give 313 mg (24%)
methyl(S)-3-nitro-4-(2-amino-1-propoxy)benzoate as a pale yellow
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.22 (d, 3H, J=6.8 Hz),
3.43-3.48 (m, 1H), 3.69-3.87 (m, 2H), 3.89 (s, 3H), 6.93-6.95 (m,
1H), 7.99-8.02 (m, 1H), 8.18-8.21 (m, 1H).
[1373] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (591 mg, 1.23
mmol), methyl (S)-3-nitro-4-(2-amino-1-propoxy)benzoate (313 mg,
1.23 mmol), and Et.sub.3N (257 mL, 1.84 mmol) in DMF (5 mL) was
stirred at room temp for 2 days. The mixture was diluted with
EtOAc, washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4,
and evaporated. The residue was purified by column chromatography
on silica-gel with 5% MeOH in CHCl.sub.3 as eluent to give 310 mg
(46%) methyl
(S)-3-nitro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetylamino]-1-propoxy]benzoate as a gum. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.28 (d, 3H, J=6.8 Hz), 2.31 (s, 3H), 3.49 (s, 2H), 3.71
(s, 3H), 3.92 (s, 3H), 4.14-4.16 (m, 2H), 4.38-4.41 (m, 1H),
5.88-5.90 (m, 1H), 6.49 (s, 1H), 6.70-6.71 (m, 1H), 6.77-6.79 (m,
1H), 7.05-7.30 (m, 4H), 7.55-7.57 (m, 1H), 8.02-8.06 (m, 2H),
8.16-8.19 (m, 1H), 8.51-8.52 (m, 1H); MS (FAB) m/z 551
(M.sup.++1).
[1374] A stirred solution of methyl
(S)-3-nitro-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamin-
o]-1-propoxy]benzoate (310 mg, 0.56 mmol) in MeOH-THF (10 mL, 1:1,
v/v) was hydrogenated over 5% Pd--C (50 mg, 16 wt %) overnight. The
mixture was filtered to remove the catalyst and the filtrate was
evaporated to give methyl
(S)-3-amino-4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetylamin-
o]-1-propoxy]benzoate (240 mg) as a gum.
[1375] To a stirred solution of the above crude product (220 mg) in
THF-MeOH (10 mL, 1:1, v/v) was added 0.5 N NaOH (10 mL), and the
resulting mixture was heated under reflux for 2 hr. The mixture was
poured into ice-H.sub.2O, and the aqueous layer was made acidic (pH
4.8) by the addition of 1 N HCl. The solid was collected and the
crude solid was recrystallized from MeOH--CHCl.sub.3-n-hexane to
give 116 mg (2 steps, 44%) 293 as a pale yellow crystalline powder.
mp 200-204.degree. C.; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.21 (d,
3H, J=6.8 Hz), 2.24 (s, 3H), 3.37 (s, 2H), 3.80 (s, 3H), 3.83-3.99
(m, 2H), 4.10-4.19 (m, 1H), 4.99 (bs, 2H), 6.75-7.23 (series of m,
total 8H), 7.79 (d, 1H, J=7.8 Hz), 7.98 (d, 1H, J=7.8 Hz), 8.17 (d,
1H, J=8.3 Hz), 8.46 (s, 1H), 8.55 (s, 1H); MS (FAB) m/z 507
(M.sup.++1); Anal. Calcd for
C.sub.27H.sub.30N.sub.4O.sub.6.1/2H.sub.2O: C, 62.90; H, 6.06; N,
10.87. Found: C, 62.85; H, 6.10; N, 10.51.
Example 243
(S)-4-[2-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetylamino]-1-propo-
xy]benzoic acid
[1376] ##STR1916##
[1377] To a stirred and cooled (0.degree. C.) solution of
(S)-2-(N-tert-butoxycarbonylamino)-1-propanol (6.74 g, 0.04 mol),
benzyl 4-hydroxybenzoate (8.78 g, 0.04 mol), and Ph.sub.3P (15.13
g, 0.06 mol) in THF (100 mL) was added diisopropyl azodicarboxylate
(DIAD) (11.4 mL, 0.06 mol), and the resulting mixture was heated
under reflux overnight. The solution was evaporated and the residue
was dissolved in CH.sub.3Cl.sub.2 (30 mL) and TFA (30 mL). The
resulting solution was stirred at room temp for 30 min., and the
solution was evaporated in vacuo. The residue was dissolved in
CHCl.sub.3, washed with sat. NaHCO.sub.3, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3 to CHCl.sub.3:MeOH
(9:1, v/v) as eluent to give 6.63 g (2 steps, 60%) benzyl
(S)-4-(2-amino-1-propoxy)benzoate as a yellow oil. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.18 (d, J=6.3 Hz, 3H), 3.35-3.39 (m, 1H),
3.71-3.75 (m, 1H), 3.90-3.93 (m, 1H), 5.34 (s, 2H), 6.90-6.93 (m,
2H), 7.32-7.46 (m, 5H), 8.01-8.04 (m, 2H).
[1378] A mixture of
3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetic acid (480 mg,
1.27 mmol), benzyl (S)-4-(2-amino]-propoxy)benzoate (361 mg, 1.27
mmol), EDC (hydrochloride) (364 mg, 1.90 mmol), HOBt (256 mg, 1.89
mmol), and 4-DMAP (31 mg, 0.25 mmol) in DMF (8 mL) was stirred at
room temp. overnight. The mixture was diluted with EtOAc, washed
with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4, and evaporated.
The residue was purified by column chromatography on silica-gel
with CHCl.sub.3 to 5% MeOH in CHCl.sub.3 as eluent to give 476 mg
(58%) benzyl
(S)-4-[2-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetylamino]-1-prop-
oxy]benzoate as a brown solid. .sup.1H-NMR (CDCl.sub.3) .delta.
1.25-1.28 (m, 3H), 3.51 (s, 2H), 3.74 (s, 3H), 3.95-3.97 (m, 2H),
4.36-4.39 (m, 1H), 5.33 (s, 2H), 6.75-6.94 (m, 5H), 7.26-7.70 (m,
8H), 7.99-8.26 (m, 5H).
[1379] To a stirred solution of benzyl
(S)-4-[2-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetylamino]-1-prop-
oxy]benzoate (476 mg, 1.39 mmol) in THF (10 mL) was added 0.5 N
NaOH (10 mL), and the resulting mixture was heated under reflux for
2 hr. The mixture was poured into ice-1 N HCl, and the solid was
collected. The crude solid was recrystallized from
MeOH--CHCl.sub.3-n-hexane to give 240 mg (59%) 294 as a white
crystalline powder. mp 202-205.degree. C.; .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.18 (d, J=6.8 Hz, 3H), 3.37 (s, 2H), 3.82
(s, 3H), 3.92-4.00 (m, 2H), 4.03-4.15 (m, 1H), 6.77-6.79 (m, 1H),
6.93-7.03 (m, 4H), 7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H), 7.87-7.97
(m, 4H), 8.13-8.15 (m, 1H), 8.73 (s, 1H), 8.91 (s, 1H), 12.63 (bs,
1H); MS (FAB) m/z 557 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.26BrN.sub.3O.sub.6: C, 56.12; H, 4.71; Br, 14.36; N,
7.55. Found: C, 56.11; H, 4.74; Br, 14.56; N, 7.49.
Example 244
(S)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2-aminobe-
nzyl)amino]-1-propoxy]benzoic acid
[1380] ##STR1917##
[1381] To a stirred cooled (0.degree. C.) solution of benzyl
(S)-4-(2-amino-1-propoxy)benzoate (1.50 g, 5.26 mmol) and
2-nitrobenzaldehyde (0.87 g, 5.76 mmol) in MeOH--AcOH (16 mL, 15:1,
v/v) was added NaBH.sub.3CN (1.65 g, 26.3 mmol), and the resulting
mixture was stirred at room temp overnight. The mixture was
quenched by sat. NaHCO.sub.3 and extracted with EtOAc. The extract
was washed with brine, dried over Na.sub.2SO.sub.4, and evaporated.
The residue was purified by column chromatography on silica-gel
with CHCl.sub.3 to 5% MeOH in CHCl.sub.3 as eluent to give 931 mg
(42%) benzyl (S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.21 (d, J=6.4 Hz,
3H), 3.13-3.18 (m, 1H), 3.88-3.97 (m, 2H), 4.06-4.20 (m, 2H), 5.34
(s, 2H), 6.89-6.94 (m, 2H), 7.29-7.65 (m, 8H), 7.94-8.03 (m, 3H);
MS (FAB) m/z 421 (M.sup.++1).
[1382] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (460 mg, 0.96
mmol), benzyl (S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate (403
mg, 0.96 mmol), and Et.sub.3N (200 mL, 1.43 mmol) in DMF (8 mL) was
stirred at room temp overnight. The mixture was diluted with EtOAc,
washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4, and
evaporated. The residue was purified by column chromatography on
silica-gel with 1% MeOH in CHCl.sub.3 as eluent to give 504 mg
(73%) benzyl
(S)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetyl]-(2-nitrob-
enzyl)amino]-1-propoxy]benzoate as a brown amorphous solid. MS
(FAB) m/z 717 (M.sup.++1).
[1383] A stirred solution of benzyl
(S)-4-[[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetyl]-(2-nitrobenzyl)amino]-1-propoxy]benzoate (504 mg, 0.70
mmol) in MeOH-THF (11 mL, 10:1, v/v) was hydrogenated over 5% Pd--C
(100 mg, 20 wt %) at 3 atm overnight. The mixture was filtered to
remove the catalyst and the filtrate was evaporated. The residue
was purified by preparative TLC with 5% MeOH in CHCl.sub.3 as
eluent to give 115 mg (27%) 295 as a white powder. MS (FAB) m/z 597
(M.sup.++1).
Example 245
(S)-4-[[2-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetyl]-(2-nitroben-
zyl)amino]-1-propoxy]benzoic acid
[1384] ##STR1918##
[1385] To a stirred and cooled (0.degree. C.) solution of benzyl
(S)-4-(2-amino-1-propoxy)benzoate (1.50 g, 5.26 mmol) and
2-nitrobenzaldehyde (0.87 g, 5.76 mmol) in MeOH--AcOH (16 mL, 15:1,
v/v) was added NaBH.sub.3CN (1.65 g, 26.3 mmol), and the resulting
mixture was stirred at room temp overnight. The mixture was
quenched by sat. NaHCO.sub.3 and extracted with EtOAc. The extract
was washed with brine, dried over Na.sub.2SO.sub.4 and evaporated.
The residue was purified by column chromatography on silica-gel
with CHCl.sub.3 to 5% MeOH in CHCl.sub.3 as eluent to give 931 mg
(42%) benzyl (S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.21 (d, 3H, J=6.4
Hz), 3.13-3.18 (m, 1H), 3.88-3.97 (m, 2H), 4.06-4.20 (m, 2H), 5.34
(s, 2H), 6.89-6.94 (m, 2H), 7.29-7.65 (m, 8H), 7.94-8.03 (m, 3H);
MS (FAB) m/z 421 (M.sup.++1).
[1386] A mixture of
3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetic acid (476 mg,
1.26 mmol), benzyl (S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate
(528 mg, 1.26 mmol), EDC (hydrochloride) (361 mg, 1.88 mmol), HOBt
(255 mg, 1.89 mmol), and DMAP (30 mg, 0.25 mmol) in DMF (10 mL) was
stirred at room temp. overnight and at 60.degree. C. for 1 day. The
mixture was diluted with EtOAc, washed with 0.5 N HCl, brine, dried
over NaSO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3 to 2% MeOH in
CHCl.sub.3 as eluent to give benzyl
(S)-4-[[2-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetyl]-2-nitroben-
zyl)amino]-1-propoxy]benzoic acid as an oil, which is used to the
subsequent reaction without further purification.
[1387] To a stirred solution of the above crude in THF-MeOH (10 mL,
1:1, v/v) was added 0.5 N NaOH (10 mL), and the resulting mixture
was heated under reflux for 3 hr. The mixture was poured into
ice-H.sub.2O, and the basic aqueous layer was made acidic (pH 4.3)
with 1 N HCl. The solid was collected, and the crude solid was
purified by preparative TLC with 5% MeOH in CHCl.sub.3 as eluent to
give 162 mg (2 steps, 19%) 296 as a white amorphous solid. MS (FAB)
m/z 692 (M.sup.++1); Anal. Calcd for
C.sub.33H.sub.31BrN.sub.4O.sub.8. 7/4H.sub.2O: C, 54.82; H, 4.81;
N, 7.75. Found: C, 54.80; H, 4.61; N, 7.24.
Example 246
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetamido]ethoxy]benz-
oic acid
[1388] ##STR1919##
[1389] To a cooled (0.degree. C.) solution of
2-(N-tert-butoxycarbonylamino)ethanol (3.20 g, 19.9 mmol), methyl
4-hydroxybenzoate (3.02 g, 19.9 mmol) and Ph.sub.3P (6.25 g, 23.8
mmol) in THF (50 ml) was added dropwise DIAD (4.69 ml, 23.8 mmol)
over for 5 min. The reaction mixture was heated under reflux for 3
hr. The mixture was evaporated and the residue was dissolved in
CH.sub.2Cl.sub.2 (30 ml) and TFA (30 ml). The reaction mixture was
stirred at room temperature overnight. The mixture was concentrated
in vacuo and the residue was dissolved in CHCl.sub.3 and H.sub.2O.
The solution was made basic by sat. NaHCO.sub.3 and extracted with
CHCl.sub.3. The extract was washed with brine, dried over
Na.sub.2SO.sub.4 and evaporated. The residue was purified by column
chromatography on silica-gel with CHCl.sub.3-MeOH (30:1, v/v) as
eluent to give methyl 4-(2-aminoethoxy)benzoate (1.03 g, 34% for 2
steps) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
3.10-3.13 (m, 2H), 3.89 (s, 3H), 4.03-4.06 (m, 2H), 6.92-6.94 (m,
2H), 7.98-8.00 (m, 2H).
[1390] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (557 mg,
1.77 mmol), methyl 4-(2-aminoethoxy)benzoate (346 mg, 1.77 mmol),
EDC.HCl (408 mg, 2.13 mmol), HOBt (287 mg, 2.12 mmol) and DMAP (52
mg, 0.43 mmol) in DMF (10 ml) was stirred at room temperature for 2
days. The mixture was diluted with EtOAc and H.sub.2O. The
resulting precipitate was collected to give methyl
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetamido]ethoxy]ben-
zoate (598 mg, 69%) as a white crystalline powder. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.23 (s, 3H), 3.36 (s, 2H), 3.42-3.45 (m,
2H), 3.79 (s, 3H), 3.81 (s, 3H), 4.02-4.09 (m, 2H), 6.73-6.75 (m,
1H), 6.90-6.94 (m, 2H), 7.02-7.04 (m, 2H), 7.09-7.15 (m, 2H),
7.77-7.79 (m, 1H), 7.88-7.90 (m, 2H), 7.95-7.98 (m, 1H), 8.23-8.24
(m, 1H), 8.44 (s, 1H), 8.53 (s, 1H); MS (FAB), m/z 492 (M.sup.++1);
Anal. Calcd for C.sub.27H.sub.29N.sub.3O.sub.6.1/4H.sub.2O: C,
65.38; H, 5.99; N, 8.47. Found: C, 65.26; H, 5.99; N, 8.49.
[1391] To a stirred solution of methyl
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetamido]ethoxy]ben-
zoate (280 mg, 0.57 mmol) in THF-MeOH (10 ml, 1:1, v/v) was added
0.5 N NaOH (10 ml) and the reaction mixture was heated under reflux
for 5 hr. The mixture was cooled to room temperature and poured
into ice-1 N HCl. The resulting precipitate was collected and
recrystallized from Et.sub.2O--CHCl.sub.3-MeOH to give 297 (135 mg,
50%) as a white crystalline powder. MW 477.51 .sup.1H-NMR
(DMSO-d.sub.6) .delta. 2.24 (s, 3H), 3.38 (s, 2H), 3.43-3.47 (m,
2H), 3.82 (s, 3H), 4.06-4.09 (m, 2H), 6.76-6.78 (m, 1H), 6.92-7.17
(m, 6H), 7.79 (d, J=8.1 Hz, 1H), 7.88-7.89 (m, 2H), 7.98 (d, J=8.1
Hz, 1H), 8.24-8.27 (m, 1H), 8.45 (s, 1H), 8.55 (s, 1H); MS (FAB)
m/z 478 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.27N.sub.3O.sub.6.1/4H.sub.2O: C, 64.79; H, 5.75; N,
8.72. Found: C, 64.67; H, 5.63; N, 8.60.
Example 247
(S)-4-[2-N-[4-[N'-2-bromophenyl)ureido]-3-methoxyphenylacetamido]-1-propox-
y]benzoic acid
[1392] ##STR1920##
[1393] To a cooled (0.degree. C.), stirred solution of
(S)-2-(N-tert-butoxycarbonylamino)-1-propanol (6.74 g, 0.04 mol),
benzyl 4-hydroxybenzoate (8.78 g, 0.04 mol), and Ph.sub.3P (15.13
g, 0.06 mol) in THF (100 ml) was added DIAD (11.4 ml, 0.06 mol),
and the reaction mixture was heated under reflux overnight. The
solution was evaporated and the residue was dissolved in
CH.sub.2Cl.sub.2 (30 ml) and TFA (30 ml). The solution was stirred
at room temperature for 30 min. and the solution was concentrated
in vacuo. The residue was dissolved in CHCl.sub.3, washed with sat.
NaHCO.sub.3, dried over Na.sub.2SO.sub.4 and evaporated. The
residue was purified by column chromatography on silica-gel with
CHCl.sub.3 to CHCl.sub.3-MeOH (9:1, v/v) as eluent to give benzyl
(S)-4-(2-amino-1-propoxy)benzoate (6.63 g, 2 steps, 60%) as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.18 (d, J=6.3 Hz,
3H), 3.35-3.39 (m, 1H), 3.71-3.75 (m, 1H), 3.90-3.93 (m, 1H), 5.34
(s, 2H), 6.90-6.93 (m, 2H), 7.32-7.46 (m, 5H), 8.01-8.04 (m,
2H).
[1394] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (480 mg,
1.27 mmol), benzyl (S)-4-(2-amino-1-propoxy)benzoate (361 mg, 1.27
mmol), EDC.HCl (364 mg, 1.90 mmol), HOBt (256 mg, 1.89 mmol) and
DMAP (31 mg, 0.25 mmol) in DMF (8 ml) was stirred at room
temperature overnight. The mixture was diluted with EtOAc, washed
with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4 and evaporated.
The residue was purified by column chromatography on silica-gel
with CHCl.sub.3 to 5% MeOH in CHCl.sub.3 as eluent to give benzyl
(S)-4-[2-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetamido]-1-propox-
y]benzoate (476 mg, 58%) as a brown solid. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.25-1.28 (m, 3H), 3.51 (s, 2H), 3.74 (s, 3H), 3.95-3.97
(m, 2H), 4.36-4.39 (m, 1H), 5.33 (s, 2H), 6.75-6.94 (m, 5H),
7.26-7.70 (m, 8H), 7.99-8.26 (m, 5H).
[1395] The a stirred solution of benzyl
(S)-4-[2-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl
acetamido]-1-propoxy]benzoate (476 mg, 1.39 mmol) in THF (10 ml)
was added 0.5 N NaOH (10 ml), and the reaction mixture was heated
under reflux for 2 hr. The mixture was poured into ice-1 N HCl, and
the resulting precipitate was collected. The crude solid was
purified by recrystallization from MeOH--CHCl.sub.3-n-hexane to
give 298 (240 mg, 59%) as a white crystalline powder. MW 556.41
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.18 (d, J=6.8 Hz, 3H), 3.37 (s,
2H), 3.82 (s, 3H), 3.92-4.00 (m, 2H), 4.03-4.15 (m, 1H), 6.77-6.79
(m, 1H), 6.93-7.03 (m, 4H), 7.30-7.34 (m, 1H), 7.59-7.61 (m, 1H),
7.87-7.97 (m, 4H), 8.13-8.15 (m, 1H), 8.73 (s, 1H), 8.91 (s, 1H),
12.63 (bs, 1H); MS (FAB) t/z 557 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.26BrN.sub.3O.sub.6: C, 56.12; H, 4.71; Br, 14.36; N,
7.55. Found: C, 56.11; H, 4.74; Br, 14.56; N, 7.49.
Example 248
(S)-4-[2-N-[[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylacetam-
ido]-1-propoxy]benzoic acid
[1396] ##STR1921##
[1397] To a cooled (0.degree. C.) solution of
(S)-2-[(N-tert-butoxycarbonyl)amino]-1-propanol (3.08 g, 17.6
mmol), methyl 4-hydroxybenzoate (2.67 g, 17.6 mmol) and Ph.sub.3P
(5.53 g, 21.1 mmol) in THF (35 ml) was added dropwise DIAD (4.15
ml, 21.1 mmol). The reaction mixture was heated under reflux
overnight. The mixture was evaporated and the residue was purified
by column chromatography on silica-gel with n-hexane-EtOAc (5:1,
v/v) as eluent to give methyl
(S)-4-[2-[(N-tert-butoxycarbonyl)amino]-1-propoxy]benzoate (1.24 g,
23%) as a white solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.30 (d,
J=6.8 Hz, 3H), 1.45 (s, 9H), 3.89 (s, 3H), 3.98-3.99 (m, 2H), 4.07
(m, 1H), 4.76 (m, 1H), 6.91-6.93 (m, 2H), 7.98-8.00 (m, 2H); MS
(FAB) m/z 310 (M.sup.++1).
[1398] To a stirred solution of methyl
(S)-4-[2-[(N-tert-butoxycarbonyl)amino]-1-propoxy]benzoate (1.24 g,
4.01 mmol) in CH.sub.2Cl.sub.2 (10 ml) was added TFA (10 ml) and
the reaction mixture was stirred at room temperature overnight. The
mixture was concentrated in vacuo, and made basic by sat.
NaHCO.sub.3. The mixture was extracted with CHCl.sub.3, washed with
brine, dried over K.sub.2CO.sub.3 and evaporated to give methyl
(S)-4-(2-amino]-propoxy)benzoate (790 mg, 94%) as a yellow oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.19 (d, J=6.7 Hz, 3H), 3.34-3.42
(m, 1H), 3.70-3.77 (m, 1H), 3.86-3.94 (series of s and m, total
4H), 6.92 (d, J=9.0 Hz, 2H), 7.98 (d, J=9.0 Hz, 2H).
[1399] To a cooled (0.degree. C.) solution of methyl
(S)-4-(2-amino-1-propoxy)benzoate (790 mg, 3.78 mmol) and Et.sub.3N
(630 ml, 4.52 mmol) in THF (10 ml) was added TFAA (640 ml, 4.53
mmol) and the reaction mixture was stirred at room temperature for
4 days. The mixture was diluted with 0.5 N HCl and extracted with
CHCl.sub.3. The extract was washed with brine, dried over
K.sub.2CO.sub.3, and evaporated. The residue was purified by
recrystallization from n-hexane-CHCl.sub.3 to give methyl
(S)-4-[2-trifluoroacetamido)-1-propoxy]benzoate (790 mg, 69%) as a
white crystalline material. .sup.1H-NMR (CDCl.sub.3) .delta. 1.42
(d, J=6.8 Hz, 3H), 3.89 (s, 3H), 4.01-4.13 (m, 2H), 4.44-4.50 (m,
1H), 6.57-6.61 (m, 1H), 6.92 (d, J=9.0 Hz, 2H), 8.00 (d, J=9.0 Hz,
2H); MS (FAB) m/z 306 (M.sup.++1); Anal. Calcd for
C.sub.13H.sub.14F.sub.3NO.sub.4: C, 51.15; H, 4.62; F, 18.67; N,
4.59. Found: C, 51.14; H, 4.60; F, 18.50; N, 4.54.
[1400] To a stirred solution of methyl
(S)-4-[2-trifluoroacetamido)-1-propoxy]benzoate (695 mg, 2.28 mmol)
and K.sub.2CO.sub.3 (630 mg, 4.56 mmol) in DMF (10 ml) was added
MeI (210 ml, 3.37 mmol) and the reaction mixture was stirred at
room temperature for 2 days. The mixture was diluted with H.sub.2O
and extracted with EtOAc. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with EtOAc--CHCl.sub.3 (1:19,
v/v) as eluent to give methyl
(S)-4-[2-[(N-methyl)trifluoroacetamido]-1-propoxy]benzoate (720 mg,
99%) as a white solid. mp 73-75.degree. C.; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.36-1.39 (m, 3H), 2.96 and 3.10 (each s,
total 3H), 3.89 (s, 3H), 3.99-4.14 (m, 2H), 4.84-4.92 (m, 1H),
6.88-6.92 (m, 2H), 7.98-8.00 (m, 2H); MS (FAB) m/z 320 (M.sup.++1);
Anal. Calcd for C.sub.14H.sub.16F.sub.3NO.sub.4: C, 52.67; H, 5.05;
F, 17.85; N, 4.39. Found: C, 52.76; H, 5.09; F, 17.53; N, 4.32.
[1401] To a stirred solution of methyl
(S)-4-[2-[(N-methyl)trifluoroacetamido]-1-propoxy]benzoate (710 mg,
2.22 mmol) in MeOH--H.sub.2O (10 ml, 1:1, v/v) was added
K.sub.2CO.sub.3 (460 mg, 3.33 mmol) and the reaction mixture was
stirred at room temperature overnight. The mixture was diluted with
H.sub.2O and extracted with EtOAc. The extract was washed with
brine, dried over Na.sub.2SO.sub.4, and evaporated to give methyl
(S)-4-[2-[(N-methylamino)-1-propoxy]benzoate (430 mg, 87%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.17 (d, J=6.6 Hz,
3H), 2.48 (s, 3H), 2.98-3.06 (m, 1H), 3.86-3.96 (series of s and m,
total 5H), 6.92 (d, J=9.0 Hz, 2H), 7.98 (d, J=9.0 Hz, 2H).
[1402] A mixture of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (605 mg,
1.93 mmol), methyl (S)-4-[2-(N-methylamino)-1-propoxy]benzoate (430
mg, 1.93 mmol), EDC.HCl (444 mg, 2.32 mmol), HOBt (313 mg, 2.32
mmol), Et.sub.3N (320 ml, 2.30 mmol) in THF (13 ml) was stirred at
room temperature overnight. The mixture was diluted with H.sub.2O
and extracted with EtOAc. The extract was washed with brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was purified by
column chromatography on silica-gel with CHCl.sub.3-MeOH (100:1 to
75:1, v/v) as eluent to give methyl
(S)-4-[2-N-[[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylaceta-
mido]-1-propoxy]benzoate (953 mg, 95%) as a white foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.12-1.13 and 1.25-1.27 (each m, total 3H),
2.27 (s, 3H), 2.84 and 2.92 (each s, total 3H), 3.63 (s, 3H), 3.67
(s, 2H), 3.71-4.05 (series of s and m, total 5H), 4.39-4.44 and
4.96-5.01 (each m, total 1H), 6.66-6.85 (m, 5H), 7.09-7.27 (m, 4H),
7.53-7.55 (m, 1H), 7.92-7.98 (m, 2H), 8.04-8.08 (m, 1H); MS (FAB)
m/z 520 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.33N.sub.3O.sub.6. 11/4H.sub.2O: C, 61.20; H, 6.82; N,
7.38. Found: C, 61.14; H, 5.86; N, 7.16.
[1403] To a stirred solution of methyl
(S)-4-[2-N-[[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylaceta-
mido]-1-propoxy]benzoate (663 mg, 1.28 mmol) in THF (5 ml) was
added 0.5 N NaOH (5 ml) and the reaction mixture was heated under
reflux for 5 hr. After cooled to room temperature, the mixture was
poured into ice-1 N HCl and the resulting precipitate was collected
under a reduced pressure. The crude solid was dissolved in
CHCl.sub.3 and evaporated. The residue was washed with Et.sub.2O to
give 299 (465 mg, 72%) as a white amorphous solid. MW 505.56
.sup.1H-NMR (DMSO-d.sub.6) .delta. 1.11-1.15 (m, 3H), 2.25 (s, 3H),
2.73 and 2.88 (each s, total 3H), 3.60-3.76 (m, 2H), 3.83 (s, 3H),
4.03-4.12 (m, 2H), 4.41-4.50 and 4.48-4.94 (each m, total 1H),
6.71-6.76 (m, 1H), 6.84-6.86 (m, 1H), 6.91-7.01 (m, 3H), 7.11-7.12
(m, 2H), 7.78-7.80 (m, 1H), 7.86-7.90 (m, 2H), 8.01-8.03 (m, 1H),
8.45 (s, 1H), 8.54 (s, 1H); MS (FAB) m/z 520 (M.sup.+30 1); Anal.
Calcd for C.sub.28H.sub.31N.sub.3O.sub.6. 3/2H.sub.2O: C, 63.15; H,
6.43; N, 7.89. Found: C, 63.09; H, 5.99; N, 7.64.
Example 249
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetamido]-2-methyl-1-
-propoxy]benzoic acid
[1404] ##STR1922##
[1405] A stirred mixture of methyl 4-hydroxybenzoate (3 g, 19.72
mmol), K.sub.2CO.sub.3 (6.8 g, 49.3 mmol), 3-chloropivalic acid
(2.9 g, 21.69 mmol) and catalytic amount of KI (200 mg) in DMF (70
ml) was heated at 100.degree. C. for 14 days under a current of
nitrogen. The mixture was poured into ice-water, and extracted with
EtOAc. The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was chromatographed
on silica gel (50 ml) with CHCl.sub.3-EtOH (10:1, v/v) as eluent to
give the title compound (1 g, 23%) as an amorphous solid.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.37 (s, 6H), 3.89 (s, 3H), 4.03
(s, 2H), 6.92 (d, J=9 Hz, 2H), 7.98 (d, J=9 Hz, 2H).
[1406] To a stirred mixture of
2,2-dimethyl-3-(4-methoxycarbonyl)phenoxypropionic acid (720 mg,
2.85 mmol) and triethylamine (0.46 ml, 3.28 mmol) in tert-BuOH (10
ml) and benzene (10 ml) was added a solution of diphenyl phosphoryl
azide (870 mg, 3.14 mmol) in benzene (3 ml) at room temperature.
The resulting mixture was heated at reflux for 20 hr. After
cooling, ice and 1N HCl (5 ml) was added to the mixture and
extracted with toluene. The extracts were washed with brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica gel (50 ml) with toluene-EtOAc (10:1,
v/v) as eluent to give methyl
4-[1-(2-amino-2-methyl)propoxy]benzoate as a gum (520 mg), which
was used to the subsequent reaction without further purification.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.41 (s, 9H), 3.89 (s, 3H), 4.04
(s, 2H), 4.69 (br s, 1H), 6.94 (dd, J=2 and 7 Hz, 2H), 7.98 (dd,
J=2 and 7 Hz).
[1407] A solution of methyl
4-[1-(2-methyl-2-tert-butoxycarbonylamino-)propoxy]benzoate (520
mg) and anisole (0.175 ml, 1.61 mmol) in CH.sub.2Cl.sub.2 (5 ml)
and TFA (3 ml) was stirred at room temperature for 18 hr. The
mixture was evaporated off. The residue was dissolved in
CH.sub.2Cl.sub.2, and the mixture was made basic by the addition of
sat. NaHCO.sub.3. Separated CH.sub.2Cl.sub.2 layer was dried over
Na.sub.2SO.sub.4 and Na.sub.2CO.sub.3, and evaporated. The residue
was chromatographed on silica gel with CHCl.sub.3-EtOH (10:1, v/v)
as eluent to give methyl 4-[1-(2-amino-2-methyl)propoxy]benzoate
(250 mg, 39% in two steps) as a gum. .sup.1H-NMR (CDCl.sub.3)
.delta. 3.75 (s, 2H), 3.89 (s, 3H), 6.93 (d, J=8.8 Hz, 2H), 7.98
(d, J=8.8 Hz, 2H).
[1408] To a stirred mixture of methyl
4-[1-(2-amino-2-methyl)propoxy]benzoate (250 mg, 1.12 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (352 mg,
1.12 mmol), 4-DMAP (165 mg, 1.34 mmol) in DMF (10 ml) was added
EDC.HCl (290 mg, 1.51 mmol) at room temperature. The resulting
mixture was stirred at room temperature for 18 hr. The mixture was
pored into ice-water. The solid was collected, washed with water
and air-dried. The crude solid was purified by silica gel column
chromatography with CHCl.sub.3-EtOH (4:1, v/v) as eluent to give
methyl
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetamido]-2-methyl--
1-propoxy]benzoate (580 mg, q.y.) as a crystalline material. IR
(KBr) 3350, 3286, 1712, 1687, 1637, 1606 cm.sup.-1; .sup.1H-NMR
(CDCl.sub.3) .delta. 1.39 (s, 6H), 2.33 (s, 3H), 3.41 (s, 2H), 3.63
(s, 3H), 3.88 (s, 3H), 4.05 (s, 2H), 5.44 (br s, 1H), 6.33 (br s,
1H), 6.79 (d, J=8.3 Hz, 2H), 7.12 (s, 1H), 7.18 (t, J=7.5 Hz, 1H),
7.53 (d, J=7.8 Hz, 1H), 7.94 (d, J=7.8 Hz, 2H), 8.14 (d, J=8.3 Hz,
1H); MS (FAB) m/z 520 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.33N.sub.3O.sub.6: C, 67.04; H, 6.40; N, 8.09. Found:
C, 66.86; H, 6.36; N, 8.22.
[1409] To a stirred solution of methyl
4-[2-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl
acetamido]-2-methyl-2-propoxy]benzoate (510 mg, 0.98 mmol) was
added 0.25N NaOH (8 ml, 2 mmol) at room temperature. The resulting
mixture was stirred at an ambient temperature for 18 hr. The
mixture was pored into ice-1N HCl (5 ml). the solid was collected,
washed with water and air-dried. The crude solid was recrystallized
from CHCl.sub.3-EtOH-Et.sub.2O to give 300 (480 mg, 97%) as fine
needles. MW 505.56 IR (KBr) n 3346, 3294, 1687, 1637, 1604
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 1.35 (s, 6H), 2.24
(s, 3H), 3.33 (s, 2H), 3.80 (s, 3H), 4.15 (s, 2H), 6.75 (d, J=8.3
Hz, 1H), 6.88 (s, 1H), 6.95-6.99 (m, 3H), 7.11-7.17 (m, 3H), 7.80
(d, J=8.3 Hz, 1H), 7.82 (s, 1H), 7.87 (d, J=8.8 Hz, 2H), 7.98 (d,
J=7.8 Hz, 1H), 8.45 (s, 1H), 8.54 (s, 1H), 12.62 (br s, 1H); MS
(FAB) m/z 506 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.31N.sub.3O.sub.6: C, 66.52; H, 6.18; N, 8.31. Found:
C, 66.22; H, 6.28; N, 8.11.
Example 250
3-amino-4-[2-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetamido]-2--
methyl-1-propoxy]benzoic acid
[1410] ##STR1923##
[1411] To a stirred solution of 4-amino-2-nitrophenol (10 g, 64.88
mmol) in AcOH (70 ml) and DMSO (20 ml) was added c. H.sub.2SO.sub.4
at 0-5.degree. C. To a stirred this solution was added dropwise a
solution of NaNO.sub.2 (5.4 g, 77.9 mmol) in water (5 ml) below 20
CC for over 10 min. The resulting mixture was further stirred for
0.5 hr at 5.degree. C. This mixture was poured into a stirred
solution of KI (30 g, 0.182 mol) and catalytic amount of Cu powder
(200 mg) in ice-water (200 ml) for over 10 min. The resulting
mixture was for a further 1 hr at an ambient temperature. The
mixture was extracted with CH.sub.2Cl.sub.2. The extracts were
washed successively with sat. Na.sub.2S.sub.2O.sub.3 and brine. The
organic layer was dried over Na.sub.2SO.sub.4, and evaporated. The
residue was chromatographed on silica-gel (50 ml) with
CHCl.sub.3-EtOAc (3:1, v/v) as eluent to give 4-iodo-2-nitrophenol
(2.5 g, 15%) as a yellow crystalline material. .sup.1H-NMR
(CDCl.sub.3) .delta. 6.94 (d, J=8.8 Hz, 1H), 7.82 (dd, J=2 and 8.8
Hz, 1H), 8.42 (d, J=2 Hz, 1H), 10.49 (s, 1H).
[1412] To a stirred solution of 4-iodo-2-nitrophenol (2 g, 7.75
mmol), hydroxypivalic acid methyl ester (1.05 g, 7.92 mmol) and
PPh.sub.3 (2.3 g, 8.68 mmol) in THF (10 ml) was added dropwise a
solution of DIAD (1.77 g, 8.30 mmol) in THF (2 ml) under ice-water
bath cooling. The resulting mixture was then heated under reflux
for 18 hr. After cooling, the mixture was evaporated off. The
residue was chromatographed on silica gel (100 ml) with
toluene-EtOAc (4:1, v/v) as eluent to give methyl
3-(4-iodo-2-nitro)phenoxy-2,2-dimethylpropionate (2.9 g, q.y.) as a
crystalline material. .sup.1H-NMR (CDCl.sub.3) .delta. 1.34 (s,
6H), 3.71 (s, 3H), 4.08 (s, 2H), 6.86 (d, J=8.8 Hz, 1H), 7.78 (dd,
J=2 and 8.8 Hz, 1H), 8.12 (d, J=2 Hz, 1H).
[1413] A mixture of methyl
3-(4-iodo-2-nitro)phenoxy-2,2-dimethylpropionate (2.8 g, 7.38 mmol)
in THF (15 ml) and 0.25 N NaOH (60 ml, 15 mol) at an ambient
temperature for 18 hr. The mixture was poured into ice-1N HCl (20
ml). The solid was collected, washed with water and air-dried. The
crude solid was recrystallized from CHCl.sub.3-EtOH--IPE to afford
3-(4-iodo-2-nitro)phenoxy-2,2-dimethylpropionic acid (2.0 g, 74%)
as a crystalline material. Mp 165-182.degree. C.; IR (KBr) n 1716,
1525, 1344 cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. 1.38 (s,
6H), 4.10 (s, 2H), 6.86 (d, J=8.8 Hz, 1H), 7.79 (dd, J=2.2 and 8.8
Hz, 1H), 8.12 (d, J=2 Hz, H); MS (FAB) m/z 366 (M.sup.++1); Anal.
Calcd for C.sub.29H.sub.33N.sub.3O.sub.6: C, 36.18; 13.18; N, 3.84.
Found: C, 36.85; H, 3.35; N, 3.79.
[1414] To a stirred mixture of
3-(4-iodo-2-nitro)phenoxy-2,2-dimethylpropionic acid (1.93 g, 5.29
mmol) and triethylamine (590 mg, 5.81 mmol) in tert-BuOH (15 ml)
and toluene (15 ml) was added a solution of diphenyl phosphoryl
azide (1.53 g, 5.55 mmol) in toluene (3 ml) at room temperature.
The resulting mixture was then heated at reflux for 20 hr. After
cooling, ice and 1N HCl (5 ml) was added to the mixture and
extracted with toluene. The extracts were washed with brine, dried
over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica gel (50 ml) with toluene-EtOAc (10:1,
v/v) as eluent to give
3-nitro-4-(2-tert-butoxycarbonylamino-2-methyl-1-propoxy)iodobenzene
(1.91 g, 83%) as a gum. .sup.1H-NMR (CDCl.sub.3) .delta.1.38 (s,
9H), 1.39 (s, 6H), 4.19 (s, 2H), 4.67 (br s, 1H), 6.88 (d, J=8.8
Hz, 1H), 7.77 (dd, J=2.0 and 8.8 Hz, 1H), 8.12 (d, J=2.0 Hz,
1H).
[1415] A mixture of
3-nitro-4-(2-tert-butoxycarbonylamino-2-methyl-1-propoxy)iodobenzene
(1.9 g, 4.36 mmol), Pd(OAc).sub.2 and
1,3-bis(diphenylphosphino)propane (dppp) (90 mg, 0.22 mmol) in
triethylamine-MeOH-DMSO (1:2:5, v/v, 48 ml) was stirred under a
current of CO (gas) at 70.degree. C. for 6 hr. After cooling, the
mixture was poured into water and extracted with EtOAc. The
extracts were washed with brine, dried over Na.sub.2SO.sub.4, and
evaporated. The residue was chromatographed on silica gel (50 ml)
with toluene-EtOAc (6:1, v/v) as eluent to give methyl
4-(2-tert-butoxycarbonyl amino-2-methyl-1-propoxy)-3-nitrobenzoate
(820 mg, 51%) as a gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.38 (s,
9H), 1.42 (s, 6H), 3.93 (s, 3H), 4.29 (s, 2H), 4.67 (br s, 1H),
7.15 (d, J=8.8 Hz, 11), 8.18 (dd, J=1.7 and 8.8 Hz, 1H), 8.52 (d,
J=1.7 Hz, 1H).
[1416] A stirred mixture of methyl
4-(2-tert-butoxycarbonylamino-2-methyl-1-propoxy)-3-nitrobenzoate
(350 mg, 0.95 mmol) and 5% Pd--C (70 mg) in EtOH (30 ml) was
hydrogenated in an atmospheric hydrogen pressure at room
temperature for 20 hr. Insoluble Pd-catalyst was removed with
suction and washed with EtOH The filtrate was evaporated off to
afford methyl 4-(2-tert-butoxycarbonyl
amino-2-methyl)-1-propoxy-3-aminobenzoate as a gum, which was used
to the subsequent reaction without further purification.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.41 (s, 9H), 1.43 (s, 6H), 3.86
(s, 3H), 4.07 (s, 2H), 4.67 (br s, 1H), 6.80 (d, J=8.5 Hz, 1H),
7.39 (d, J=2.2 Hz, 1H), 7.44 (dd, J=2.2 and 8.5 Hz, 1H).
[1417] To a stirred mixture of the above methyl
4-(2-tert-butoxycarbonylamino-2-methyl)-1-propoxy-3-aminobenzoate
and triethylamine (0.20 ml, 1.43 mmol) in CH.sub.2Cl.sub.2 (10 ml)
was added a solution of trifluoroacetic anhydride (0.182 ml, 1.28
mmol) in CH.sub.2Cl.sub.2 (3 ml) at 0-5.degree. C. The resulting
mixture was stirred at room temperature for 1 hr. Ice-sat.
NaHCO.sub.3 was added to the mixture, and extracted with
CH.sub.2Cl.sub.2. The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and evaporated. The residue was dissolved in
CH.sub.2Cl.sub.2 (5 ml) and added anisole (0.105 ml, 0.95 mmol) and
TFA (2 ml). The resulting mixture was stirred at room temperature
for 18 hr. The mixture was evaporated in vacuo, and the residue was
diluted with CH.sub.2Cl.sub.2 and made basic by the addition of
sat. NaHCO.sub.3. The separated CH.sub.2Cl.sub.2 layer was dried
over Na.sub.2SO.sub.4, and evaporated. The residue was
chromatographed on silica gel (50 ml) with CHCl.sub.3-EtOH (99:1,
v/v) as eluent to give methyl
4-(2-amino-2-methyl-1-propoxy)-3-trifluoroacetamidobenzoate (631
mg, 63% in 3-steps) as a gum. .sup.1H-NMR (CDCl.sub.3) .delta. 1.29
(s, 9H, tert-Bu), 3.86 (s, 2H, CH.sub.2), 3.91 (s, 3H,), 6.99 (d,
J=8.5 Hz, 1H), 7.91 (dd, J=2.0 and 8.5 Hz, 1H), 8.83 (d, J=8.5 Hz,
1H).
[1418] To a stirred mixture of methyl
4-(2-amino-2-methyl)-1-propoxy-3-trifluoroacetamido benzoate (200
mg, 0.598 mmol),
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (210 mg,
0.598 mmol), 4-DMAP (90 mg, 0.72 mmol) in DMF (7 ml) was added
EDC.HCl (160 mg, 0.81 mmol) at room temperature. The resulting
mixture was stirred at room temperature for 20 hr. The mixture was
pored into ice-water. The solid was collected, washed with water
and air-dried. The crude solid was purified by silica gel column
chromatography with CHCl.sub.3-EtOH (98:2, v/v) as eluent to give
methyl
4-[2-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetamido]-2-methyl--
1-propoxy]-3-trifluoroacetamidobenzoate (580 mg, q.y.) as a
crystalline material. .sup.1H-NMR (CDCl.sub.3) .delta. 1.42 (s,
6H), 3.42 (s, 2H), 3.69 (s, 3H), 3.89 (s, 3H), 4.23 (s, 2H), 5.33
(br s, 1H), 6.66 (s, 1H), 6.71 (m, 1H), 6.92 (d, J=8.5 Hz, 1H),
7.02 (m, 1H), 7.09 (m, 2H), 7.29 (m, 1H), 7.37 (d, J=8.0 Hz, 1H),
7.87 (dd, J=2 and 8.5 Hz, 1H), 7.97 (d, J=8.0 Hz, 1H), 8.19 (d,
J=8.2 Hz, 1H), 8.59 (br s, 1H), 8.74 (d, J=2.0 Hz, 1H).
[1419] To a stirred solution of methyl
4-[2-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl
acetamido]-2-methyl-1-propoxy]-3-trifluoroacetamidobenzoate (320
mg, 0.492 mmol) in THF (2 ml) was added 0.25N NaOH (6 ml, 1.5 mmol)
at an ambient temperature. And the resulting mixture was stirred
for 20 hr. The mixture was poured into ice-1N HCl (2 ml). The solid
was collected, washed with water and air-dried. The crude solid was
recrystallized from CHCl.sub.3-EtOH-Et.sub.2O to give 301 (240 mg,
89%) as fine needles. MW 541.00 IR (KBr) n 3338, 3296, 1691, 1641
cm.sup.-1; .sup.1H-NMR (CDCl.sub.3) .delta. .sup.1H-NMR
(DMSO-d.sub.6) .delta. 1.37 (s, 6H), 3.35 (s, 2H), 3.77 (s, 3H),
4.05 (s, 2H), 4.96 (br s, 1H), 6.75 (br d, J=8.3 Hz, 1H), 6.78 (d,
J=8.3 Hz, 1H), 6.87 (d, J=1.7 Hz, 1H), 7.01 (m, 1H), 7.15 (dd, J=2
and 8.5 Hz, 1H), 7.24 (d, J=2 Hz, 1H), 7.27 (dt, J=2.0 and 8.5 Hz,
1H), 7.43 (dd, J=2 and 8.0 Hz, 1H), 7.78 (br s, 1H), 7.90 (d, J=8.0
Hz, 1H), 8.08 (dd, J=2 and 8.3 Hz, 1H), 8.85 (s, 1H), 8.89 (s, 1H),
12.23 (br s, 1H); MS (FAB) m/z 541 (M.sup.++1); Anal. Calcd for
C.sub.27H.sub.29ClN.sub.4O.sub.6: C, 58.00; H, 5.59; N, 10.02.
Found: C, 57.97; H, 5.39; N, 10.01.
Example 251
2-acetylamino-4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-met-
hylacetamido]ethylaminobenzoateic acid
[1420] ##STR1924##
[1421] To a stirred solution of 2-acetylamino-4-nitrobenzoic acid
(1.28 g, 5.71 mmol) in benzene-MeOH (4:1, v/v, 25 mL), was added
trimethylsilyldiazomethane (2.0 M solution in n-hexane, 4.28 ml,
8.56 mmol) at 0.degree. C. The stirring was continued for 18 hours
at rt. The reaction was poured into hexane, and the resulting
precipitate was collected by filtration to give methyl
2-acetylamino-4-nitrobenzoate (1.32 g, 97%) as a white solid; mp no
data; .sup.1H-NMR (400 MHz, CDCl.sub.3) .delta. 2.30 (s, 3H), 4.00
(s, 3H), 7.88 (m, 1H), 8.18 (dd, J=2.0 Hz, 8.8 Hz, 1H), 9.60 (t,
J=2.2 Hz, 1H), 11.10 (s, 1H); MS (ESI) m/z 238 (M.sup.+).
[1422] To a solution of methyl 2-acetylamino-4-nitrobenzoate (1.31
g, 5.50 mmol) in MeOH (30 mL) was added 5% Pd on carbon (195 mg),
and the stirring under H.sub.2 gas (3 atm) was continued for 18
hours at rt. The catalyst was filtered off and the mixture was
evaporated. The resulting crude solid was recrystallized with
CHCl.sub.3-MeOH-hexane to give methyl 2-acetylamino-4-aminobenzoate
(1.03 g, 90%) as a white solid. .sup.1H-NMR (400 MHz, CDCl.sub.3)
.delta. 2.23 (s, 3H), 3.82 (s, 3H), 4.20 (s, 2H), 6.30 (dd, J=2.5
Hz, 8.8 Hz, 1H), 7.80 (d, J=8.8 Hz, 1H), 8.06 (s, 1H), 11.26 (s,
1H); MS (FAB), m/z 208 (M.sup.+).
[1423] To a cooled solution of methyl 2-acetylamino-4-aminobenzoate
(300 mg, 1.44 mmol) and N-tert-butoxycarbonyl-N-methylglycinal (499
mg, 2.88 mmol) in 1,2-dichloroethane (30 ml), was added
NaBH(OAc).sub.3 (964 mg, 4.32 mmol) and the stirring was continued
for 64 h at 0.degree. C. The mixture was poured into sat.
NaHCO.sub.3 and was extracted with CHCl.sub.3 (50 ml.times.3),
washed with brine, and dried over MgSO.sub.4. After removal of the
solvent in vacuo, the residue was chromatographed on silica gel
(middle pressure chromatography system: YAMAZEN YFLC-5404-FC,
linear gradient of hexane-EtOAc from 9:1 to 2:1) to give methyl
2-acetylamino-4-[2-(N-tert-butoxycarbonyl-N-methylamino)ethylamino]benzoa-
te (451 mg, 86%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3, 400
MHz) .delta. 1.48 (s, 9H), 2.22 (s, 3H), 2.90 (s, 3H), 3.32 (m,
2H), 3.50 (m, 2H), 3.80 (s, 3H), 6.20 (dd, J=2.2 Hz, 8.8 Hz, 1H),
7.80 (m, 1H), 7.95 (m, 1H), 11.30 (brs, 1H); MS (FAB) m/z 366
(M.sup.++1).
[1424] To a stirred solution of methyl
2-acetylamino-4-[2-(N-tert-butoxycarbonyl-N-methylamino)ethylamino]benzoa-
te (450 mg, 1.23 mmol) in dichloromethane (5 mL), was added TFA (5
mL) and the stirring was continued for 18 h at rt. After removal of
the solvent in vacuo, the residue was dissolved in CHCl.sub.3 (200
mL), washed with brine, sat. NaHCO.sub.3, and dried over
MgSO.sub.4. The solvent was removed to give methyl
2-acetylamino-4-[2-(N-methylamino)ethylamino]benzoate (298 mg, 88%)
as a colorless oil. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.21
(s, 3H), 2.46 (s, 3H), 2.88 (m, 2H), 3.31 (m, 2H), 3.83 (s, 3H),
4.85 (br, 1H), 6.24 (dd, J=2.5 Hz, 8.8 Hz, 1H), 7.80 (d, J=8.8 Hz,
1H), 7.99 (d, J=2.5 Hz, 1H); MS (FAB), m/z 266 (M.sup.++1).
[1425] A mixture of methyl
2-acetylamino-4-[2-(N-methylamino)ethylamino]benzoate (145 mg, 0.55
mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid
(172 mg, 0.55 mmol), EDC.HCl (158 mg, 0.83 mmol), HOBt (141 mg,
1.05 mmol), and DMAP (13 mg, 0.11 mmol) in DMF (10 ml) was stirred
for 18 hours. The mixture was diluted with EtOAc (300 ml), washed
with brine, and dried over MgSO.sub.4. After removal of the
solvent, residue was chromatographed on silica gel (middle pressure
chromatography system: YAMAZEN YFLC-5404-FC, linear gradient of
CHCl.sub.3-MeOH from 100:0 to 70:30) to give methyl
2-acetylamino-4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-me-
thylacetamido]ethylaminobenzoate (309 mg, 100%) as an amorphous
foam. .sup.1H-NMR (CDCl.sub.3) .delta. 2.22 (s, 3H), 2.30 (s, 3H),
3.02 (s, 3H), 3.35 (m, 2H), 3.58 (s, 3H), 3.50-3.74 (m, 4H), 3.85
(s, 3H), 6.20 (n, 1H), 6.58 (s, 1H), 6.65-6.75 (m, 3H), 7.13 (m,
2H), 7.40-7.50 (m, 2H), 7.75 (m, 2H), 7.90 (m, 1H), 8.00 (m, 1H),
11.32 (s, 1H); MS (FAB) m/z 562 (M.sup.++1).
[1426] To a solution of methyl
2-acetylamino-4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-me-
thylacetamido]ethylaminobenzoate (309 mg, 0.55 mmol) in THF-MeOH
(1:1, v/v, 9 ml), was added 0.25 N NaOH (4.4 ml, 1.1 mmol) at rt,
and heated to reflux. The stirring was continued for 18 hours at
reflux. The reaction mixture was poured into water, and acidified
to pH 5.0 with 1.0 N HCl. The resulting precipitate was
recrystallized with hexane-diethylether to give 302 (175 mg, 58%)
as a pale red powder. MW 547.60 .sup.1H-NMR (CD.sub.3OD) .delta.
2.16 (d, J=4.8 Hz, 3H), 2.28 (d, J=4.2 Hz, 3H), 2.98 and 3.10 (2 s,
total 3H), 3.35 (m, 2H), 3.68 (m, 4H), 3.81 and 3.84 (2 s, total
3H), 6.30 (m, 1H), 6.60-6.82 (m, 2H), 7.00 (m, 1H), 7.15 (m, 2H),
7.53 (m, 1H), 7.77-7.96 (m, 3H); MS (FAB) m/z 548 (M.sup.++1);
Anal. Calcd for C.sub.29H.sub.33N.sub.5O.sub.6.0.5H.sub.2O: C,
62.58; H, 6.16; N, 12.58. Found: C, 62.55; H, 6.31; N, 12.15.
Example 252
2-acetylamino-4-[2-N-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl]-N-meth-
ylacetamido]ethylaminobenzoic acid
[1427] ##STR1925##
[1428] A mixture of methyl
2-acetylamino-4-(2-N-methylamino-1-ethylamino)benzoate (145 mg,
0.55 mmol), 3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetic acid
(209 mg, 0.55 mmol), EDC.HCl (158 mg, 0.83 mmol), HOBt (141 mg,
1.05 mmol), and DMAP (13 mg, 0.11 mmol) in DMF (10 ml) was stirred
for 18 hours. The mixture was diluted with EtOAc (300 ml), washed
with brine, and dried over MgSO.sub.4. After removal of the
solvent, residue was chromatographed on silica gel (middle pressure
chromatography system: YAMAZEN YFLC-5404-FC, linear gradient of
CHCl.sub.3-MeOH from 100:0 to 70:30) to give methyl
2-acetylamino-4-[2-N-[4-[N'-(2-bromo
phenyl)ureido]-3-methoxyphenyl]-N-methylacetamido]ethylaminobenzoate
(294 mg, 85%) as an amorphous foam. .sup.1H-NMR (CDCl.sub.3)
.delta. 2.21 (s, 3H), 3.05 (s, 3H), 3.40 (m, 2H), 3.65-3.70 (m,
4H), 3.78 (s, 3H), 3.86 (s, 3H), 6.21 (m, 1H), 6.79 (m, 2H), 6.93
(m, 1H), 7.10 (d, J=10.3 Hz, 1H), 7.30 (m, 1 h), 7.42 (m, H), 7.61
(m, 1H), 7.78-7.85 (m, 2H), 7.93 (m, 2H), 8.13 (m, 1H); MS (FAB),
m/z 627 (W).
[1429] To a solution of methyl
2-acetylamino-4-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenylacetamido]-2-
-N-methylamino-1-ethylaminobenzoate (294 mg, 0.47 mmol) in THF-MeOH
(1:1, v/v, 8 ml), was added 0.25 N NaOH (3.8 ml, 0.94 mmol) at rt,
and heated to reflux. The stirring was continued for 18 hours at
reflux. The reaction mixture was poured into water, and acidified
to pH 5.0 with 1.0 N HCl. The resulting precipitate was
recrystallized with hexane-diethylether to give 303 as a white
powder (210 mg, 73%). MW 612.47 mp 155-160.degree. C.; .sup.1H-NMR
(CD.sub.3OD) .delta. 22.18 (d, J=5.5 Hz, 3H), 3.00 and 3.12 (2 s,
total 3H), 3.39 (m, 1H), 3.60 (m, 4H), 3.70 (s, 1H), 3.85 and 3.86
(2 s, total 3H), 6.31 (m, 1H), 6.68 (m, 1H), 6.78 (m, 1H), 6.78 and
6.85 (2 m, total 1H), 6.97 (m, 1H), 7.30 (m, 1H), 7.56 (m, 1H),
7.80-7.95 (m, 4H); MS (ESI) m/z 613 (M.sup.+); Anal. Calcd for
C.sub.28H.sub.31Br.sub.1N.sub.5O.sub.6.0.75H.sub.2O: C, 53.64; H,
5.22; N, 11.17. Found: C, 53.89; H, 5.23; N, 10.69.
Example 253
4-[2-N-[[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]-N-phenylacetamido]-
ethoxy]benzoic acid
[1430] ##STR1926##
[1431] To a solution of methyl
4-[2-methanesulfonyloxy)ethoxy]benzoate (2.74 g, 10 mmol) in MeCN
(50 ml), was added aniline (9.1 ml, 100 mmol) at rt. The reaction
was stirred for 64 hours at reflux. The mixture was poured into
H.sub.2O (200 mL), extracted with EtOAc (100 mL.times.2), dried
over MgSO.sub.4. After removal of the solvent in vacuo, the
unreacted aniline was removed in vacuo by co-evaporation with
toluene (10 mL.times.3) at 80.degree. C. The residue was
chromatographed on silica gel (middle pressure chromatography
system: YAMAZEN YFLC-5404-FC, f50 mm.times.150 mm, CHCl.sub.3) to
give methyl 4-[2-(N-phenylamino)ethoxy]benzoate (2.23 g, 82%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 3.56 (t, J=5.1 Hz,
2H), 3.90 (s, 3H), 4.21 (t, J=5.1 Hz, 2H), 6.68 (dd, J=1.0 Hz, 8.6
Hz, 2H), 6.75 (t, J=7.3 Hz, 1H), 7.20 (AB type d, J=7.3 Hz, 2H),
8.00 (d, J=9.1 Hz, 2H); MS (ESI) m/z 272 (M.sup.++1).
[1432] A mixture of methyl 4-[2-(N-phenylamino)ethoxy]benzoate (136
mg, 0.5 mmol), 3-methoxy-4-[N'-(2-chlorophenyl)ureido]phenylacetic
acid (167 mg, 0.5 mmol) and PyBOP (781 mg, 0.75 mmol),
i-PrNEt.sub.2 (261 ml, 0.96 mmol) in DMF (10 ml) was stifled for 18
hours. The mixture was diluted with EtOAc (100 mL), washed with 1 N
HCl, brine and dried over MgSO.sub.4. The residue was co-evaporated
with toluene (10 ml.times.3) to remove DMF. The residue was
chromatographed on TLC (MERCK, silica gel 60, 2 mm, 2 plates,
CHCl.sub.3-MeOH, 20:1) to give methyl
4-[2-N-[[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]-N-phenylacetamido-
]ethoxy]benzoate (129 mg, 44%) as a white amorphous foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 3.42 (s, 1H), 3.69 (d, J=8.3 Hz,
1H), 3.74 (s, 3H), 3.87 (s, 3H), 4.10 (m, 2H), 4.23 (m, 2H),
6.48-7.44 (m, 13H), 7.93 (d, J=9.3 Hz, 2H), 8.18 (dd, J=1.5 Hz, 8.3
Hz, 1H); MS (ESI) m/z 588 (M.sup.+).
[1433] To a solution of methyl
4-[2-N-[[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]-N-phenyl
acetamido]ethoxy]benzoate (124 mg, 0.19 mmol) in THF-MeOH (6:1,
v/v, 3 ml), was added 0.5 N NaOH (2 ml, 1 mmol) at rt, and heated
to reflux in a sealed bottle. The stirring was continued for 15
hours at reflux. The reaction mixture was poured into water,
acidified with 1.0 N HCl, extracted with CHCl.sub.3-MeOH (2:1, 20
mL.times.3), and dried over MgSO.sub.4. After removal of the
solvent, the residue was crystallized with
CHCl.sub.3-hexane-diethylether to give 304 (77 mg, 64%) as a white
powder. MW 574.02 .sup.1H-NMR (CD.sub.3OD) .delta. 3.45 (s, 2H),
3.79 (s, 3H), 4.12 (m, 2H), 4.22 (m, 2H), 6.48 (dd, J=2.0 Hz, 8.3
Hz, 1H), 6.61 (d, J=2.0 Hz, 1H), 6.87 (d, J=8.8 Hz, 2H), 7.00 (m,
1H), 7.22 (m, 3H), 7.36 (m, 1H), 7.43 (m, 3H), 7.90 (d, J=8.3 Hz,
1H), 7.95 (d, J=8.8 Hz, 2H), 8.02 (dd, J=1.5 Hz, 8.3 Hz, 1H); MS
(ESI) m/z 574 (Mt); Anal. Calcd for
C.sub.31H.sub.28ClN.sub.3O.sub.6.0.5H.sub.2O: C, 63.86; H, 5.01; N,
7.21. Found: C, 63.67; H, 4.91; N, 6.99.
Example 254
(S)-4-[2-N-[[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-(2-aminobenz-
yl)acetamido]-1-propoxy]benzoic acid
[1434] ##STR1927##
[1435] To a cooled (0.degree. C.) solution of benzyl
(S)-4-(2-amino-1-propoxy)benzoate (1.50 g, 5.26 mmol) and
2-nitrobenzaldehyde (0.87 g, 5.76 mmol) in MeOH--AcOH (16 ml, 15:1,
v/v) was added NaBH.sub.3CN (1.65 g, 26.3 mmol) and the reaction
mixture was stirred at room temperature overnight. The mixture was
quenched by sat. NaHCO.sub.3 and extracted with EtOAc. The extract
was washed with brine, dried over Na.sub.2SO.sub.4 and evaporated.
The residue was purified by column chromatography on silica gel
with CHCl.sub.3 to 5% MeOH in CHCl.sub.3 as eluent to give benzyl
(S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate (931 mg, 42%) as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.21 (d, J=6.4 Hz,
3H), 3.13-3.18 (m, 1H), 3.88-3.97 (m, 2H), 4.06-4.20 (m, 2H), 5.34
(s, 2H), 6.89-6.94 (m, 2H), 7.29-7.65 (m, 8H), 7.94-8.03 (m, 3H);
MS (FAB) m/z 421 (M.sup.++1).
[1436] A mixture of pentafluorophenyl
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetate (460 mg, 0.96
mmol), benzyl (S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate (403
mg, 0.96 mmol) and Et.sub.3N (200 ml, 1.43 mmol) in DMF (8 ml) was
stirred at room temperature overnight. The mixture was diluted with
EtOAc, washed with 0.5 N HCl, brine, dried over Na.sub.2SO.sub.4,
and evaporated. The residue was purified by column chromatography
on silica-gel with 1% MeOH in CHCl.sub.3 as eluent to give benzyl
(S)-4-[2-N-[[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-(2-nitroben-
zyl)acetamido]-1-propoxy]benzoate (504 mg, 73%) as a brown
amorphous solid. MS (FAB), m/z 717 (M.sup.++1).
[1437] A stirred solution of benzyl
(S)-4-[2-N-[[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-(2-nitroben-
zyl)acetamido]-1-propoxy]benzoate (504 mg, 0.70 mmol) in MeOH-THF
(11 ml, 10:1, v/v) was hydrogenated over 5% Pd--C (100 mg, 20 wt %)
at 3 atm overnight. The mixture was filtered to remove the catalyst
and the filtrate was evaporated. The residue was purified by
preparative thin layer chromatography with 5% MeOH in CHCl.sub.3 as
eluent to give 305 (115 mg, 27%) as a white powder. MW 596.67 MS
(FAB), m/z 597 (M.sup.++1).
Example 255
(S)-4-[2-N-[[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl]-N-(2-nitrobenzy-
l)acetamido]-1-propoxy]benzoic acid
[1438] ##STR1928##
[1439] To a cooled (O) solution of benzyl
(S)-4-(2-amino-1-propoxy)benzoate (1.50 g, 5.26 mmol) and
2-nitrobenzaldehyde (0.87 g, 5.76 mmol) in MeOH--AcOH (16 ml, 15:1,
v/v) was added NaBH.sub.3CN (1.65 g, 26.3 mmol) and the reaction
mixture was stirred at room temperature overnight. The mixture was
quenched by sat. NaHCO.sub.3 and extracted with EtOAc. The extract
was washed with brine, dried over Na.sub.2SO.sub.4 and evaporated.
The residue was purified by column chromatography on silica gel
with CHCl.sub.3 to 5% MeOH in CHCl.sub.3 as eluent to give benzyl
(S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate (931 mg, 42%) as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.21 (d, J=6.4 Hz,
3H), 3.13-3.18 (m, 1H), 3.88-3.97 (m, 2H), 4.06-4.20 (m, 2H), 5.34
(s, 2H), 6.89-6.94 (m, 2H), 7.29-7.65 (m, 8H), 7.94-8.03 (m, 3H);
FAB-MAS, m/z 421 (M.sup.++1).
[1440] A mixture of
4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic acid (476 mg,
1.26 mmol), benzyl (S)-4-[2-(2-nitrobenzylamino)-1-propoxy]benzoate
(528 mg, 1.26 mmol), EDC.HCl (361 mg, 1.88 mmol), HOBt (255 mg,
1.89 mmol) and DMAP (30 mg, 0.25 mmol) in DMF (10 ml) was stirred
at room temperature overnight. And the reaction could not be
completed, so the reaction mixture was stirred at 60.degree. C. for
1 day. The mixture was diluted with EtOAc, washed with 0.5 N HCl,
brine, dried over Na.sub.2SO.sub.4 and evaporated. The residue was
purified by column chromatography on silica-gel with CHCl.sub.3 to
2% MeOH in CHCl.sub.3 as eluent to give the title compound as a
crude oil. To a stirred solution of the crude product in THF-MeOH
(10 ml, 1:1, v/v) was added 0.5 N NaOH (10 ml) and the reaction
mixture was heated under reflux for 3 hr. The mixture was poured
into ice-H.sub.2O and the basic aqueous layer was acidified (pH
4.3) with 1 N HCl. The resulting precipitate was collected and the
crude solid was purified by preparative thin layer chromatography
with 5% MeOH in CHCl.sub.3 as eluent to give 306 (162 mg, 2 steps,
19%) as a white amorphous solid. MW 691.53 MS (FAB), m/z 692
(M.sup.++1); Anal. Calcd for C.sub.33H.sub.31BrN.sub.4O.sub.8.
7/4H.sub.2O: C, 54.82; H, 4.81; N, 7.75. Found: C, 54.80; H, 4.61;
N, 7.24.
Example 256
4-[2-N-cyclopropyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]acetam-
ido]ethoxybenzoic acid
[1441] ##STR1929##
[1442] A mixture of methyl 4-(2-cyclopropylaminoethoxy)benzoate
(290 mg, 1.23 mmol),
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenylacetic acid (412 mg,
1.23 mmol), EDC.HCl (354 mg, 1.85 mmol), HOBt (cat.), and DMAP
(cat.) in DMF (10 ml) was stirred overnight. The mixture was
partitioned between EtOAc (300 ml) and H.sub.2O (100 ml). The
organic phase was separated, washed with brine (2.times.100 ml),
dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1) as eluent
to give methyl
4-[2-N-cyclopropyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]aceta-
mido]ethoxybenzoate (506 mg, 75%) as a yellow viscous oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 0.90-0.97 (m, 4H), 2.75 (m, 1H),
3.61 (s, 3H), 3.79 (t, J=5.4 Hz, 2H), 3.87 (s, 3H), 3.88 (s, 2H),
4.16 (t, J=5.4 Hz, 2H), 6.76-6.80 (m, 4H), 6.95 (dt, J=7.8, 1.5 Hz,
1H), 7.21-7.31 (m, 2H), 7.53 (s, 1H), 7.56 (s, 1H), 7.93 (d, J=8.3
Hz, 3H), 8.19 (dd, J=8.3, 1.5 Hz, 1H).
[1443] To a stirred solution of methyl
4-[2-N-cyclopropyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]aceta-
mido]ethoxybenzoate (506 mg, 0.917 mmol) in THF (7 ml) was added
0.25 N NaOH (7.3 ml, 1.83 mmol). After stirring overnight, the
mixture was poured into 1 N HCl (50 ml) and extracted with
CHCl.sub.3-MeOH (4:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1 to 10:1)
as eluent to give 307 (403 mg, 82%) as a colorless amorphous solid.
MW 537.99 .sup.1H-NMR (DMSO) .delta. 0.86-0.91 (m, 4H), 2.75 (m,
1H), 3.69 (t, J=5.5 Hz, 2H), 3.81 (s, 3H), 3.84 (s, 2H), 4.16 (t,
J=5.5 Hz, 2H), 6.76 (d, J=8.3 Hz, 1H), 6.88 (s, 1H), 6.97-7.04 (m,
3H), 7.28 (t, J=7.8 Hz, 1H), 7.44 (d, J=7.8 Hz, 1H), 7.88 (d, J=8.8
Hz, 2H), 7.96 (d, J=8.1 Hz, 1H), 8.10 (d, J=8.3 Hz, 1H), 8.89 (s,
1H), 8.93 (s, 1H), 12.65 (s, br s); MS (FAB), m/z 538 (M.sup.++1);
Anal. Calcd for C.sub.28H.sub.28ClN.sub.3O.sub.6: C, 62.51; H,
5.25; N, 7.81. Found: C, 61.85; H, 5.42; N, 7.41.
Example 257
4-[2-N-cyclohexyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetami-
do]ethoxybenzoic acid
[1444] ##STR1930##
[1445] To a solution of methyl
4-[(2-methanesulfonyloxy)-1-ethoxy]benzoate (2.74 g, 10 mmol) in
MeCN (50 ml), was added cyclohexylamine (5.72 ml, 50 mmol) at rt
The reaction was stirred for 18 hours at reflux. The mixture was
poured into H.sub.2O (200 mL), extracted with EtOAc (100
mL.times.2), dried over MgSO.sub.4. After removal of the solvent,
residue was chromatographed on silica gel (middle pressure
chromatography system: YAMAZEN YFLC-5404-FC, f50 mm.times.150 mm,
linear gradient of CHCl.sub.3-EtOAc from 10:0 to 1:1) to give
methyl 4-(2-N-cyclohexylamino)ethoxy benzoate (2.43 g, 88%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.10 (m, 2H), 1.25
(m, 2H), 1.60 (br, 2H), 1.73 (m, 2H), 1.90 (br, 2H), 2.49 (m, 1H),
3.02 (t, J=5.2 Hz, 2H), 3.88 (s, 3H), 4.12 (t, J=5.2 Hz, 2H), 6.90
(d, J=6.90 Hz, 2H), 7.99 (d, J=7.99 Hz, 2H); MS (ESI) m/z 278
(M.sup.++1).
[1446] A mixture of methyl 4-(2-N-cyclohexylamino)ethoxybenzoate
(139 mg, 0.5 mmol),
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (157 mg,
0.5 mmol), EDC.HCl (144 mg, 0.75 mmol), HOBt (128 mg, 0.95 mmol),
and DMAP (12 mg, 0.1 mmol) in DMF (2.5 ml) was stirred for 18
hours. The mixture was diluted with EtOAc (200 ml), washed with 1N
HCl and brine, and dried over MgSO.sub.4. After removal of the
solvent, residue was chromatographed on silica gel (middle pressure
chromatography system: YAMAZEN YFLC-5404-FC, linear gradient
CHCl.sub.3-EtOAc 10:0 to 1:4) to give methyl
4-[2-N-cyclohexyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetam-
ido]ethoxybenzoate (247 mg, 86%) as an amorphous foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 1.08-1.80 (m, 10H), 2.30 (s, 3H), 3.60-3.79
(m, 8H), 3.88 (s, 3H), 4.16 (m, 2H), 6.30 (s, 1H), 6.70-6.83 (m,
2H), 6.88 (d, 2H, J=9.0 Hz), 7.12 (m, 2H), 7.23 (m, 1H), 7.60 (d,
1H, J=8.3 Hz), 7.92 (d, 2H, J=9.0 Hz), 8.10 (d, 1H, J=8.0 Hz); MS
(ESI) m/z 574 (M.sup.++1).
[1447] To a solution of methyl
4-[2-N-cyclohexyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetam-
ido]ethoxybenzoate (247 mg, 0.43 mmol) in THF-MeOH (6:1, v/v, 7
ml), was added 0.5 N NaOH (3.4 ml, 0.84 mmol) at rt, and heated to
reflux in a sealed bottle. The stirring was continued for 18 hours
at reflux. The reaction mixture was poured into water, acidified
with 1.0 N HCl, extracted with CHCl.sub.3-MeOH (2:1, 20
mL.times.3), and dried over MgSO.sub.4. After removal of the
solvent, the residue was crystallized with
CHCl.sub.3-hexane-diethylether to give 308 (196 mg, 81%) as a white
powder. MW 559.62 .sup.1H-NMR (CD.sub.3OD) .delta. 0.90-1.82 (m,
10H), 2.29 (s, 3H), 3.62 (m, 2H), 3.78 (s, 3H), 3.80 (m, 3H), 4.12
(m, 2H), 6.82 (m, 2H), 6.96 (m, 3H), 7.16 (m, 2H), 7.58 (d, J=7.7
Hz, 1H), 7.92 (m, 3H); MS (ESI) m/z 560 (M.sup.++1); Anal. Calcd
for C.sub.32H.sub.37N.sub.3O.sub.6.0.5H.sub.2O: C, 67.59; H, 6.74;
N, 7.39. Found: C, 67.83; H, 6.80; N, 7.13.
Example 258
4-[2-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]-N-propargylacetamid-
o]ethoxybenzoic acid
[1448] ##STR1931##
[1449] To a solution of methyl
4-[(2-methanesulfonyloxy)-1-ethoxy]benzoate (2.74 g, 10 mmol) in
MeCN (50 ml), was added propargylamine (3.43 ml, 50 mmol) at rt.
The reaction was stirred for 18 hours at reflux. The mixture was
poured into H.sub.2O (200 mL), extracted with EtOAc (100
mL.times.2), dried over MgSO.sub.4. After removal of the solvent in
vacuo, the residue was chromatographed on silica gel (middle
pressure chromatography system: YAMAZEN YFLC-5404-FC, f50
mm.times.150 mm, linear gradient of CHCl.sub.3-EtOAc from 10:0 to
9:1) to give methyl 4-(2-N-propargylamino)ethoxybenzoate (2.33 g,
100%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.28 (d,
J=2.4 Hz, 1H), 3.11 (t, J=5.1 Hz, 2H), 3.52 (d, J=2.4 Hz, 2H), 3.88
(s, 3H), 4.15 (t, J=5.1 Hz, 2H), 6.90 (d, J=8.8 Hz, 2H), 7.98 (d,
J=8.8 Hz, 2H); MS (ESI) m/z 234 (M.sup.++1).
[1450] A mixture of methyl 4-(2-N-propargylamino)ethoxybenzoate
(117 mg, 0.5 mmol),
3-methoxy-4-[N'-(2-chlorophenyl)ureido]phenylacetic acid (167 mg,
0.5 mmol), EDC.HCl (144 mg, 0.75 mmol), HOBt (128 mg, 0.96 mmol),
and DMAP (12 mg, 0.1 mmol) in DMF (10 ml) was stirred for 18 hours.
The mixture was diluted with EtOAc (100 mL), washed with 1 N HCl,
brine and dried over MgSO.sub.4. The residue was coevaporated with
toluene (10 ml.times.3) to remove DMF. The residue was
chromatographed on TLC (MERCK, silica gel 60, 2 mm, 2 plates,
CHCl.sub.3-MeOH, 20:1) to give methyl
4-(2-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]-N-propargyl
acetamido]ethoxybenzoate (244 mg, 89%) as a white amorphous foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 2.20 and 2.32 (2 m, total 1H),
3.72 (s, 2H), 3.83 (m, 5H), 3.88 (s, 3H), 4.09-4.35 (m, 4H),
6.77-6.86 (m, 4H), 6.99 (m, 1H), 7.11 (m, 2H), 7.24 (m, 1H), 7.34
(d, J=7.9 Hz, 1H), 7.96 (m, 3H), 8.18 (dd, J=1.5 Hz, 8.3 Hz, 1H);
MS (ESI) m/z 550 (M.sup.+).
[1451] To a solution of methyl
4-[2-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]-N-propargyl
acetamido]ethoxybenzoate (240 mg, 0.44 mmol) in THF-MeOH--H.sub.2O
(2:2:1, v/v, 10 ml), was added NaOH (500 mg, 12.5 mmol) at rt. The
stirring was continued for 2 hours at rt. The reaction mixture was
poured into water, acidified with 1.0 N HCl, extracted with
CHCl.sub.3-MeOH (2:1, 20 mL.times.3), and dried over MgSO.sub.4.
The residue was chromatographed on TLC (Whatman, 1 mm, 3 plates,
CHCl.sub.3-MeOH, 92:8) to give 309 (202 mg, 86%) as a white solid.
MW 535.98 .sup.1H-NMR (CD.sub.3OD) .delta. 2.60 and 2.81 (2d, J=2.5
Hz, total 1H), 3.79-3.94 (m, 4H), 3.85 (s, 3H), 4.15 (m, 1H), 4.24
(m, 1H), 4.32 (m, 2H), 6.80 (d, J=8.3 Hz, 1H), 6.85 (d, J=4.3 Hz,
1H), 6.94 (m, 2H), 7.02 (m, 1H), 7.25 (m, 1H), 7.38 (m, 1H),
7.87-8.02 (m, 4H); MS (ESI) m/z 536 (M.sup.++1); Anal. Calcd for
C.sub.2, H.sub.26ClN.sub.3O.sub.6.2.25H.sub.2O: C, 58.33; H, 5.33;
N, 7.29. Found: C, 58.23; H, 4.77; N, 6.91.
Examples 259 and 260
4-[2-N-allyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]acetamido]et-
hoxybenzoic acid
[1452] ##STR1932##
[1453] A mixture of methyl 4-(2-N-allylamino)ethoxybenzoate (118
mg, 0.5 mmol), 3-methoxy-4-[N'-(2-chlorophenyl)ureido]phenylacetic
acid (167 mg, 0.5 mmol), EDC.HCl (144 g, 0.75 mmol), HOBt (128 mg,
0.95 mmol), and DMAP (12 mg, 0.1 mmol) in DMF (2.5 ml) was stirred
for 18 hours. The mixture was diluted with EtOAc (300 ml), washed
with 1N HCl and brine, and dried over MgSO.sub.4. After removal of
the solvent, residue was chromatographed on silica gel (middle
pressure chromatography system: YAMAZEN YFLC-5404-FC, linear
gradient CHCl.sub.3-EtOAc 100:0 to 85:15) to give methyl
4-[2-N-allyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]acetamido]e-
thoxybenzoate (253 mg, 92%) as an amorphous foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 3.65-3.85 (m, 4H), 3.73 (s, 3H), 3.88 (s, 3H),
5.08 (m, 2H), 4.22 (m, 2H), 5.10-5.24 (m, 2H), 5.76 (m, 1H), 6.77
(m, 2H), 6.85 (m, 2H), 6.99 (m, 1H), 7.06 (m, 2H), 7.26 (m, 1H),
7.34 (d, 1H, J=8.1 Hz), 7.94 (d, 2H, J=8.8 Hz), 7.98 (m, 1H), 8.18
(d, 1H, J=6.9 Hz); MS (FAB) m/z 552 (M.sup.+).
[1454] To a solution of methyl
4-[2-N-allyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]acetamido]e-
thoxybenzoate (250 mg, 0.45 mmol) in THF-MeOH (1:1, v/v, 8 ml), was
added 0.25 N NaOH (3.6 ml, 0.91 mmol) at rt, and heated to reflux.
The stirring was continued for 18 hours at reflux. The reaction
mixture was poured into water, and acidified with 1.0 N HCl. The
resulting precipitate was collected by filtration. The precipitate
was recrystallized with hexane-diethylether to give 310 as a white
powder (195 mg, 80%). MW 537.99 .sup.1H-NMR (CD.sub.3OD) .delta.
3.61 (s, 1H), 3.76 (s, 3H), 3.82 (m, 1H), 3.85 (s, 1H), 3.88 (m,
1H), 4.11-4.25 (m, 4H), 5.12-5.25 (m, 2H), 5.81 (m, 1H), 6.78 (d,
1H, J=8.3 Hz), 6.82 (m, 1H), 6.92 (m, 2H), 7.01 (m, 1H), 7.26 (m,
1H), 7.37 (m, 1H), 7.96 (m, 3H), 8.02 (m, 1H); MS (FAB) m/z 537
(W); Anal. Calcd for C.sub.28H.sub.28ClN.sub.3O.sub.6.1/4H.sub.2O:
C, 61.99; H, 5.30; N, 7.75. Found: C, 62.00; H, 5.56; N, 7.76.
Example 261
4-[2-N-allyl-N-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl]acetamido]eth-
oxybenzoic acid
[1455] ##STR1933##
[1456] A mixture of methyl 4-(2-N-allylamino)ethoxybenzoate (118
mg, 0.5 mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic
acid (190 mg, 0.5 mmol), EDC.HCl (144 mg, 0.75 mmol), HOBt (128 mg,
0.95 mmol), and DMAP (12 mg, 0.1 mmol) in DMF (2.5 ml) was stirred
for 18 hours. The mixture was diluted with EtOAc (300 ml), washed
with 1N HCl and brine, and dried over MgSO.sub.4. After removal of
the solvent, residue was chromatographed on silica gel (middle
pressure chromatography system: YAMAZEN YFLC-5404-FC, linear
gradient CHCl.sub.3-EtOAc 100:0 to 70:30) to give methyl
4-[2-N-allyl-N-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl]acetamido]et-
hoxybenzoate (251 mg, 84%) as an amorphous foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 3.65-3.85 (m, 4H), 3.73 (s, 3H), 3.88 (s, 3H),
4.08 (m, 2H), 4.22 (m, 2H), 5.10-5.25 (m, 2H), 5.78 (m, 1H), 6.79
(m, 1H), 6.85 (m, 3H), 6.93 (m, 1H), 7.02 (m, 2H), 7.30 (m, 1H),
7.51 (m, 1H), 7.94 (d, 2H J=8.8 Hz), 7.97 (m, 1H), 8.14 (m, 1H); MS
(FAB) m/z 596 (M.sup.+).
[1457] To a solution of methyl
4-[2-N-allyl-N-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl]acetamido]et-
hoxybenzoate (251 mg, 0.42 mmol) in THF-MeOH (1:1, v/v, 8 ml), was
added 0.25 N NaOH (3.4 ml, 0.84 mmol) at rt, and heated to reflux.
The stirring was continued for 18 hours at reflux. The reaction
mixture was poured into water, and acidified with 1.0 N HCl. The
resulting precipitate was collected by filtration. The precipitate
was recrystallized with Hexane-diethylether to give 311 (192 mg,
78%) as a white powder. MW 582.44 .sup.1H-NMR (CD.sub.3OD) .delta.
3.61 (s, 3H), 3.77 (s, 3H), 3.80 (m, 1H), 3.85 (s, 1H), 3.88 (s,
1H), 4.12-4.25 (m, 4H), 5.12-5.23 (m, 2H), 5.81 (m, 1H), 6.76 (m,
1H), 6.82 (m, 1H), 6.93 (m, 3H), 7.29 (m, 1H), 7.56 (m, 1H), 7.94
(m, 4H); MS (FAB), m/z 582 (M.sup.+); Anal. Calcd for
C.sub.28H.sub.28BrN.sub.3O.sub.6: C, 57.74; H, 4.85; N, 7.21.
Found: C, 57.40; H, 5.07; N, 7.04. For HCl salt of 311: Anal. Calcd
for C.sub.28H.sub.27BrN.sub.3O.sub.6.0.25H.sub.2O: C, 55.23; H,
4.55; N, 6.90. Found: C, 54.98; H, 4.71; N, 6.53.
Example 262
4-[2-N-allyl-N-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]eth-
oxybenzoic acid
[1458] ##STR1934##
[1459] A mixture of methyl 4-(2-N-allylamino-1-ethyl)ethoxybenzoate
(87 mg, 0.37 mmol),
3-methyl-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (100 mg,
0.37 mmol), EDC.HCl (105 mg, 0.56 mmol), HOBt (95 mg, 0.70 mmol),
and DMAP (9 mg, 0.07 mmol) in DMF (7.4 ml) was stiffed for 18
hours. The mixture was diluted with EtOAc (100 ml), washed with 1N
HCl and brine, and dried over MgSO.sub.4. The residue was
co-evaporated with toluene (10 ml.times.3) to remove DMF. The
residue was chromatographed on TLC (Whatman, PLK-5F, 2 plates,
CHCl.sub.3-MeOH, 97:3) to give methyl
4-[2-N-allyl-N-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]et-
hoxybenzoate (190 mg, 100%) as a white amorphous foam. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.05 and 2.09 (2s, total 3H), 2.20 and 2.21
(s, total 3H), 3.62 (s, 2H), 3.76 (m, 2H), 3.90 (s, 3H), 3.88 (m,
1H), 4.08 (m, 2H), 4.11 (m, 1H), 6.28 (m, 2H), 5.78 (m, 1H), 6.88
(d, J=8.8 Hz, 2H), 7.05 (m, 1H), 7.12 (m, 1H), 7.21 (m, 1H), 7.58
(m, 2H), 7.95 (d, J=8.8 Hz, 2H), 8.02 (m, 1H); MS (FAB), m/z 516
(M.sup.++1).
[1460] To a solution of methyl
4-[2-N-allyl-N-[3-methyl-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]et-
hoxybenzoate (217 mg, 0.43 mmol) in THF-MeOH (6:1, v/v, 7 ml), was
added 0.5 N NaOH (1.9 ml, 0.86 mmol) at rt, and heated to reflux.
The stirring was continued for 2 hours at reflux in a sealed
bottle. The reaction mixture was poured into water, acidified with
1.0 N HCl, extracted with CHCl.sub.3-MeOH (2:1, 20 mL.times.3), and
dried over MgSO.sub.4. After removal of the solvent, the residue
was crystallized with CHCl.sub.3-hexane-diethylether to give 312
(84 mg, 38%) as a white powder. MW 501.57 .sup.1H-NMR (CD.sub.3OD)
.delta. 2.18 and 2.24 (s, total 3H), 2.30 (d, J=4.9 Hz, 3H), 3.70
(s, 1H), 3.78 (m, 2H), 3.88 (s, 1H), 4.12 (m, 4H), 5.20 (m, 2H),
5.81 (m, 1H), 6.92-7.20 (m, 7H), 7.58 (m, 2H), 7.96 (m, 2H); MS
(ESI) m/z 502 (M.sup.+); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.5: C, 69.44; H, 6.23; N, 8.38. Found:
C, 68.99; H, 6.39; N, 8.03.
Example 263
4-[2-N-allyl-N-[3-chloro-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]eth-
oxybenzoic acid
[1461] ##STR1935##
[1462] To a stirred solution of methyl
4-(2-N-allylaminoethoxy)benzoate (141 mg, 0.60 mmol) and
3-chloro-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (191 mg,
0.60 mmol) in DMF (5 mL) were added EDC.HCl (172.5 mg, 0.90 mmol),
HOBt (154 mg, 1.14 mmol), and DMAP (15 mg, 0.12 mmol), and the
stirring was continued overnight at rt. The mixture was diluted
with EtOAc (50 mL) and washed with 1M HCl (.times.3), 1M NaOH
(.times.1), and brine. The mixture was dried over anhydrous
MgSO.sub.4 and concentrated under a reduced pressure to give methyl
4-[2-N-allyl-N-[3-chloro-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]et-
hoxybenzoate (350 mg, 109%) as a white powder. .sup.1H-NMR
(CDCl.sub.3) .delta. 2.35 (s, 3H), 3.60 (s, 1H), 3.75 (m, 2H), 3.90
(s, 3H), 4.10 (m, 4H), 4.21 (m, 1H), 5.20 (m, 2H), 5.80 (m, 1H),
6.50 (s, 1H), 6.85 (m, 2H), 7.08 (m, 2H), 7.20 (m, 4H), 7.50 (d,
J=8.1 Hz, 1H), 7.95 (d, J=8.1 Hz, 2H), 8.12 (d, J=8.1 Hz, 1H); MS
(ESI) m/z: 536 (M.sup.++H).
[1463] To a stirred solution of methyl
4-[2-N-allyl-N-[3-chloro-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]et-
hoxybenzoate (321 mg, 0.6 mmol) in THF-MeOH--H.sub.2O (2:2:1, v/v,
30 ml), was added NaOH (500 mg, 12.5 mmol) at rt. The stirring was
continued for 18 hours at rt. The reaction mixture was poured into
water, washed with diethyl ether, acidified with 1M HCl, extracted
with CHCl.sub.3-MeOH (2:1, 20 mL.times.3), dried over anhydrous
MgSO.sub.4, and concentrated under a reduced pressure. The residue
was solidified with CHCl.sub.3/n-hexane to give 313 (283 mg, 83%)
as a white solid. IR (KBr): 3345, 1581, 1529, 1243, 1167 cm.sup.-1;
.sup.1H-NMR (CD.sub.3OD) .delta. 2.30 (s, 3H), 3.71 (s, 1H), 3.78
(m, 1H), 3.82 (m, 1H), 3.89 (s, 1H), 4.10 (m, 1H), 4.19 (m, 2H),
4.21 (t, J=5.4 Hz, 1H), 5.20 (m, 2H), 5.82 (m, 1H), 6.95 (m, 2H),
7.03 (m, 1H), 7.18 (m, 3H), 7.28 (s, 1H), 7.60 (d, J=8.1 Hz, 1H),
7.99 (m, 3H); MS (ESI) m/z 522 (M.sup.++1); Anal. Calcd for
C.sub.28H.sub.28ClN.sub.3O.sub.5.1.75H.sub.2O: C, 60.76; H, 5.74;
N, 7.59. Found: C, 60.43; H, 5.34; N, 7.17.
Example 264
methyl
4-[2-N-[2-(4-morpholinyl)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)-
ureido]phenyl]acetamido]ethoxybenzoate
[1464] ##STR1936##
[1465] To a stirred solution of methyl
4-[2-N-(2,3-dihydroxy-1-propyl)-[3-methoxy-4-[N'-(2-methylphenyl)ureido]p-
henylacetamido]ethoxy]benzoate (1.83 g, 3.24 mmol) in
THF-MeOH--H.sub.2O (1:1:1, v/v/v, 15 mL) was added sodium periodate
(2.08 g, 9.71 mmol), and stirred for 18 hours at rt. A saturated
Na.sub.2S.sub.2O.sub.3 (50 ml) was added to the reaction mixture
and the mixture was stirred for 1 hour. The mixture was extracted
with EtOAc (100 ml.times.3), washed with brine, dried over
MgSO.sub.4. The solvent was removed to give the title compound
(1.73 g, 100%) as an amorphous foam. .sup.1H-NMR (CDCl.sub.3)
.delta. 2.32 (t, 3H, J=2.8 Hz), 3.33-4.30 (m, 8H), 3.72 (s, 3H),
3.86 (s, 3H), 6.20 (m, 1H), 6.70 (m, 1H), 6.80 (m, 4H), 7.06 (m,
1H), 7.18 (m, 1H), 7.26 (m, 1H), 7.49 (d, 1H, J=7.4 Hz), 7.96 (m,
2H), 8.10 (m, 1H), 9.57 and 9.63 (2 s, total 1H); MS (FAB), m/z 534
(M.sup.+30 1).
[1466] To a stirred solution of methyl
4-[2-N-formylmethyl-N-[3-methoxy-4-[N'-(2-methyl
phenyl)ureido]phenylacetamido]ethoxy]benzoate (400 mg, 0.75 mmol)
in EtOH (7.5 ml), were added morpholine (654 ml, 7.5 mmol) and
acetic acid (429 ml, 7.5 mmol) at rt. The reaction was stirred for
5 min. at rt, then cooled to 0.degree. C. To the cooled solution,
was added NaBH.sub.3CN (471 mg, 7.5 mmol) and the stirring was
continued for 1 h at rt. The mixture was poured into sat.
NaHCO.sub.3 and was extracted with EtOAc (50 ml.times.3), washed
with brine, and dried over MgSO.sub.4. After removal of the solvent
in vacuo, the residue was chromatographed on silica gel (middle
pressure chromatography system: YAMAZEN YFLC-5404-FC, linear
gradient toluene-acetone 100:0 to 1:1) to give 314 (346 mg, 76%) as
a white amorphous foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta.
2.31 (s, 3H), 2.46 (m, 4H), 3.52-3.79 (m, 12H), 3.70 (d, 3H, J=2.7
Hz), 4.05 and 4.22 (m, total 2H), 6.24 and 6.29 (s, total 1H), 6.73
(m, 2H), 6.85 (m, 2H), 7.07 (s, 1H), 7.17 (m, 1H), 7.25 (m, 2H),
7.50 (t, 1H, J=7.3 Hz), 7.96 (m, 2H), 8.08 (m, 1H); MS (FAB), m/z
605 (M.sup.++1); Anal. Calcd for C.sub.33H.sub.40N.sub.4O.sub.7
1/2H.sub.2O: C, 64.58; H, 6.73; N, 9.13. Found: C, 64.95; H, 6.88;
N, 8.82. HCl salt of 314: Anal. Calcd for
C.sub.33H.sub.41ClN.sub.4O.sub.7.2.5H.sub.2O: C, 57.76; H, 6.76; N,
8.16; Cl, 5.17; Found: C, 58.29; H, 6.81; N, 7.42; Cl, 5.05.
Example 265
4-[2-N-[2-(4-morpholinyl)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]-
phenyl]acetamido]ethoxybenzoic acid
[1467] ##STR1937##
[1468] To a solution of methyl
4-[2-N-[2-(4-morpholinyl)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido-
]phenyl]acetamido]ethoxybenzoate (146 mg, 0.24 mmol) in THF-MeOH
(1:1, v/v, 6 ml), was added 0.25 N NaOH (1.9 ml, 0.48 mmol) at rt,
and heated to reflux. The stirring was continued for 18 hours at
reflux. The reaction mixture was poured into water, and acidified
with 1.0 N HCl. The mixture was extracted with CHCl.sub.3-MeOH
(3:1, v/v, 30 ml.times.5). The combined organic solvent was dried
over MgSO.sub.4. After removal of solvent, the residue was
crystallized with diethylether to give 315 (102 mg, 71%) as a white
powder. .sup.1H-NMR (CD.sub.3OD) .delta. 2.28 (d, J=3.0 Hz, 3H),
2.46 (m, 1H), 2.40 (m, 1H), 2.56 (m, 1H), 2.63 (m, 1H), 3.62-3.80
(m, 12H), 3.85 (s, 3H), 4.12 (m, 1H), 4.26 (m, 1H), 6.82 (m, 2H),
6.96 (m, 2H), 7.01 (m, 1H), 7.17 (m, 2H), 7.58 (d, J=7.8 Hz, 1H),
7.93 (m, 3H); MS (FAB) m/z 591 (M.sup.+); Anal. Calcd for
C.sub.32H.sub.38N.sub.4O.sub.7 1.0H.sub.2O: C, 63.14; H, 6.62; N,
9.20. Found: C, 63.48; H, 6.66; N, 8.79.
Example 266
4-[2-N-cyclopropyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetam-
ido]ethoxybenzoic acid
[1469] ##STR1938##
[1470] To a stirred solution of methyl
4-(2-hydroxyethyloxy)benzoate (5.00 g, 25.5 mmol), DMSO (18.1 ml,
255 mmol), Et.sub.3N (17.7 ml, 127.5 mmol) in CH.sub.2Cl.sub.2 (200
ml) was added SO.sub.3.Py (12.2 g, 76.5 mmol). After stirring for 5
h, the mixture was concentrated in vacuo and the residue was
diluted with H.sub.2O (100 ml). The mixture was extracted with
EtOAc (2.times.200 ml). The combined extracts were washed with
brine (100 ml), dried over (MgSO.sub.4), and evaporated. The
residue was chromatographed on silica gel with hexane-EtOAc (4:1)
to give 4:1 mixture of methyl 4-formyl methyloxybenzoate and methyl
4-hydroxybenzoate (2.00 g) as a white solid. .sup.1H-NMR
(CDCl.sub.3) .delta. 3.90 (s, 3H), 4.64 (d, J=1.0 Hz, 2H), 6.92 (d,
J=9.0 Hz, 2H), 8.02 (d, J=9.0 Hz, 2H), 9.86 (d, J=1.0 Hz, 1H).
[1471] To a stirred solution of 4:1 mixture of methyl
4-formylmethyloxybenzoate and methyl 4-hydroxybenzoate (1.00 g) and
cyclopropylamine (425 ml, 6.18 mmol) in MeOH--AcOH (10:1, 11 ml)
was added NaBH.sub.3CN (681 mg, 10.3 mmol). After stirring
overnight, the mixture was quenched by addition of sat. NaHCO.sub.3
(50 ml) and extracted with CHCl.sub.3 (2.times.200 ml). The
combined extracts were dried over MgSO.sub.4 and evaporated. The
residue was chromatographed on silica gel with CHCl.sub.3-MeOH
(20:1) to give methyl 4-(2-cyclopropyl aminoethoxy)benzoate (595
mg, 49%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
0.37-0.49 (m, 4H), 1.91 (m, 1H), 2.18-2.23 (m, 1H), 3.11 (t, J=5.2
Hz, 2H), 3.88 (s, 3H), 4.12 (t, J=5.2 Hz, 2H), 6.92 (d, J=8.8 Hz,
2H), 7.98 (d, J=8.8 Hz, 2H).
[1472] A mixture of methyl 4-(2-cylopropylaminoethoxy)benzoate (290
mg, 1.23 mmol), 3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic
acid (387 mg, 1.23 mmol), EDC.HCl (354 mg, 1.85 mmol), HOBt (cat.),
and DMAP (cat.) in DMF (10 ml) was stirred overnight. The mixture
was partitioned between EtOAc (300 ml) and H.sub.2O (100 ml). The
organic phase was separated, washed with brine (2.times.100 ml),
dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (50:1) to give
the title compound (426 mg, 65%) as a yellow viscous oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 0.90-0.97 (m, 4H), 2.28 (s, 3H),
3.60 (s, 3H), 3.77 (t, J=5.4 Hz, 2H), 3.85 (s, 2H), 3.87 (s, 3H),
4.15 (t, J=5.4 Hz, 2H), 6.60-6.81 (m, 5H), 7.09-7.23 (m, 4H), 7.57
(d, J=8.3 Hz, 1H), 7.92-7.95 (m, 2H), 8.04 (d, J=8.3 Hz, 1H).
[1473] To a stirred solution of methyl
4-[2-[N-cyclopropyl-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetam-
ido]ethoxy]benzoate (426 mg, 0.801 mmol) in THF (7 ml) was added
0.25 N NaOH (6.4 ml, 1.60 mmol). After stirring overnight, the
mixture was poured into 1 N HCl (50 ml) and extracted with
CHCl.sub.3-MeOH (4:1, 2.times.200 ml). The combined extracts were
dried over MgSO.sub.4 and evaporated. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (20:1 to 10:1)
as eluent to give 316 (333 mg, 80%) as a colorless amorphous solid.
.sup.1H-NMR (DMSO) .delta. 0.86-0.91 (m, 4H), 2.25 (s, 3H), 2.74
(m, 1H), 3.69 (t, J=5.5 Hz, 2H), 3.81 (s, 3H), 3.83 (s, 2H), 4.15
(t, J=5.5 Hz, 2H), 6.75 (d, J=8.3 Hz, 1H), 6.87 (s, 1H), 6.92-6.99
(m, 3H), 7.11-7.17 (m, 2H), 7.81 (d, J=8.1 Hz, 1H), 7.88 (d, J=8.5
Hz, 2H), 8.01 (d, J=8.1 Hz, 1H), 8.47 (s, 1H), 8.55 (s, 1H), 12.96
(s, br s); MS (FAB), m/z 518 (M.sup.++1); Anal. Calcd for
C.sub.29H.sub.31N.sub.3O.sub.6: C, 67.30; H, 6.04; N, 8.12. Found:
C, 66.71; H, 6.26; N, 7.82.
Example 267
4-[2-N-[2-(N',N'-dimethylamino)-1-ethyl)-N-[-4-[N''-(2-chlorophenyl)ureido-
]-3-methoxyphenyl]acetamido]ethoxybenzoic acid
[1474] ##STR1939##
[1475] To a solution of methyl
4-[2-N-[2-(N',N'-dimethylamino)-1-ethyl)-N-[-4-[N''-2-chlorophenyl)ureido-
]-3-methoxyphenyl]acetamido]ethoxybenzoate (100 mg, 0.17 mmol) in
THF-MeOH (1:1, v/v, 3 ml), was added 0.25 N NaOH (2.0 ml, 0.5 mmol)
at rt, and heated to reflux. The stirring was continued for 3 hours
at reflux. The reaction mixture was poured into water, and
acidified to pH 5.0 with 1.0 N HCl. After removal of the organic
solvent in vacuo, the resulting mixture was chromatographed with
HP-20 (H.sub.2O-MeOH 100:0 to 0:100) to give 317 (63 mg, 64%) as a
white powder. .sup.1H-NMR (CD.sub.3OD) .delta. 2.41 (s, 2H), 2.65
(s, 3H), 2.69 (s, 3H), 3.02 (m, 2H), 3.62-3.85 (m, 4H), 3.84 (s,
3H), 4.05 and 4.22 (m, total 2H), 6.82-6.88 (m, 4H), 7.02 (m, 1H),
7.25 (m, 1H), 7.38 (m, 1H), 7.92 (m, 2H), 8.01 (m, 2H); MS (FAB),
m/z 569 (M.sup.+); Anal. Calcd for
C.sub.29H.sub.33ClN.sub.4O.sub.6.3.0H.sub.2O: C, 55.90; H, 6.31; N,
8.99. Found: C, 56.40; H, 6.50; N, 8.08.
Example 268
isopropyl
4-[2-N-[2-(4-morpholinyl)ethyl]-N-[3-methoxy-4-[N'-(2-methylphen-
yl)ureido]phenyl]acetamido]ethoxybenzoate
[1476] ##STR1940##
[1477] To a stirred solution of
4-[2-N-[2-(4-morpholinyl)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido-
]phenyl]acetamido]ethoxybenzoic acid (250 mg, 0.42 mmol) in DMF (2
mL), were added isopropyl iodide (264 ml, 2.53 mmol) and
K.sub.2CO.sub.3 (88 mg, 0.64 mmol) at rt. The reaction was stirred
for 2 hours at 50.degree. C. The mixture was poured into brine and
was extracted with CHCl.sub.3 (50 ml.times.3), washed with brine,
and dried over MgSO.sub.4. After removal of the solvent in vacuo,
the residue was chromatographed on silica gel (middle pressure
chromatography system: YAMAZEN YFLC-5404-FC, linear gradient
toluene-acetone 100:0 to 40:60) to give 318 (261 mg, 97%) as a
white amorphous foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta.
1.35 (s, 3H), 1.36 (s, 3H), 2.31 (s, 3H), 2.45 (m, 4H), 2.50 (m,
2H), 3.55-3.78 (m, 10H), 3.70 (s, 3H), 4.05 and 4.20 (t, J=5.2 Hz,
total 2H), 5.22 (m, 1H), 6.24 and 6.33 (2 s, total 1H), 6.70-6.83
(m, 4H), 7.08 (s, 1H), 7.15 (m, 1H), 7.22 (m, 1H), 7.40 (t, J=9.0
Hz, 1H), 7.93 (d, J=8.8 Hz, 1H), 7.97 (d, J=8.8 Hz, 1H), 8.07 (t,
J=7.8 Hz, 1H); MS (FAB), m/z 633 (M.sup.++1); Anal. Calcd for
C.sub.35H.sub.44N.sub.4O.sub.7-0.75H.sub.2O: C, 65.05; H, 7.10; N,
8.67. Found: C, 65.19; H, 7.09; N, 8.50.
Example 269
4-[2-N-[2-(3,3-difluoro-1-pyrrolidinyl)ethyl]-N-[4-[N'-(2-bromophenyl)urei-
do]-3-methoxyphenyl]acetamido]ethoxybenzoic acid sodium salt
[1478] ##STR1941##
[1479] To a stirred solution of methyl
4-[2-N-formylmethyl-N-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenyl]aceta-
mido]ethoxybenzoate (300 mg, 0.5 mmol) in 1,2-dichloroethane (3.6
ml), was added 3,3-difluoropyrrolidine AcOH salt (420 mg, 2.5 mmol)
at rt. The reaction was stirred for 5 min. at rt, then cooled to
0.degree. C. To the cooled solution, was added NaBH(OAc).sub.3 (530
mg, 2.5 mmol), and the stirring was continued for 4 h at rt. The
mixture was poured into sat. NaHCO.sub.3, was extracted with
CHCl.sub.3 (50 mL.times.3), washed with brine, and dried over
MgSO.sub.4. After removal of the solvent in vacuo, the residue was
chromatographed on silica gel (middle pressure chromatography
system: YAMAZEN YFLC-5404-FC, linear gradient CHCl.sub.3-MeOH 10:0
to 97:3) to give methyl
4-[2-N-[2-(3,3-difluoro-1-pyrrolidinyl)ethyl]-N-[4-[N'-(2-bromophenyl)ure-
ido]-3-methoxyphenyl]acetamido]ethoxybenzoate (345 mg, 100%) as a
white amorphous foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta.
2.20-2.80 (m, 6H), 3.48 (m, 2H), 3.70-3.80 (m, 9H), 3.89 (s, 3H),
4.09 (m, 1H), 4.21 (m, 1H), 6.79-6.85 (m, 4H), 6.93 (m, 1H), 7.08
(m, 2H), 7.30 (m, 1H), 7.50 (m, 1H), 7.96 (m, 3H), 8.13 (dd, J=1.5
Hz, 8.3 Hz, 1H); MS (ESI) m/z 689 (M.sup.+).
[1480] To a solution of methyl
4-[2-N-[2-(3,3-difluoro-1-pyrrolidinyl)ethyl]-N-[4-[N'-(2-bromophenyl)ure-
ido]-3-methoxyphenyl]acetamido]ethoxybenzoate (345 mg, 0.5 mmol) in
THF-MeOH (1:1, v/v, 8 ml), was added 0.25 N NaOH (4 ml, 1.0 mmol)
at rt, and heated to reflux. The stirring was continued for 6 hours
at reflux. The solvent was removed, and the residue was
chromatographed on HP-20 (H.sub.2O-MeOH, 0:100 to 100:0) to give
319 (306 mg, 91%) as a pale red powder. .sup.1H-NMR (CD.sub.3OD,
400 MHz) .delta. 2.20-2.90 (m, 6H), 3.60 (m, 2H), 3.70-3.92 (m,
9H), 4.10 (m, 1H), 4.22 (m, 1H), 6.84 (m, 4H), 6.96 (m, 1H), 7.30
(m, 1H), 7.55 (m, H), 7.93 (m, 3H), 7.97 (m, 1H); MS (ESI) m/z 676
(M.sup.++1); Anal. Calcd for
C.sub.31H.sub.32BrF.sub.2N.sub.4O.sub.6.2.5H.sub.2O: C, 52.85; H,
5.29; N, 7.95. Found: C, 52.67; H, 5.20; N, 8.11.
Example 270
4-[2-N--(N'-methoxy-N'-methylamino)ethyl-N-[3-methoxy-4-[N''-(2-methylphen-
yl)ureido]phenyl]acetamido]ethoxybenzoic acid sodium salt
[1481] ##STR1942##
[1482] To a stirred solution of methyl
4-[2-N-formylmethyl-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetami-
do]ethoxybenzoate (350 mg, 0.66 mmol) in EtOH (13 ml), was added
N-methoxy-N-methylamine hydrochloride (637 mg, 6.6 mmol) at rt. The
reaction was sonicated for 5 min. at rt, then cooled to 0.degree.
C. To the cooled solution, was added NaBH.sub.3CN (105 mg, 1.65
mmol) and the stirring was continued for 18 h at rt. The mixture
was poured into sat. NaHCO.sub.3 and was extracted with CHCl.sub.3
(50 ml.times.3), washed with brine, and dried over MgSO.sub.4.
After removal of the solvent in vacuo, the residue was
chromatographed on silica gel (middle pressure chromatography
system: YAMAZEN YFLC-5404-FC, linear gradient toluene-acetone 9:1
to 2:3) to give the title compound (344 mg, 91%) as a white
amorphous foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.29 (s,
3H), 2.52 and 2.58 (s, total 3H), 2.79 (m, 2H), 3.49-3.76 (m, 9H),
3.88 (s, 3H), 4.05 and 4.20 (m, total 2H), 6.69-6.86 (m, 5H), 7.10
(m, 1H), 7.20 (m, 2 h), 7.29 (m, 1H), 7.44 (m, 1H), 7.94 and 7.99
(d, J=8.6 Hz, total 2H), 8.06 (m, 1H); MS (FAB), m/z 579
(M.sup.++1); Anal. Calcd for
C.sub.31H.sub.38N.sub.4O.sub.7.2.5H.sub.2O: C, 59.70; H, 6.95; N,
8.98. Found: C, 59.58; H, 6.65; N, 8.90.
[1483] To a solution of methyl
4-[2-N--(N'-methoxy-N'-methylamino)ethyl-N-[3-methoxy-4-[N''-(2-methylphe-
nyl)ureido]phenyl]acetamido]ethoxybenzoate (138 mg, 0.24 mmol) in
THF-MeOH (1:1, v/v, 4 ml), was added 0.25 N NaOH (1.9 ml, 0.48
mmol) at rt, and heated to reflux. The stirring was continued for
18 hours at reflux. The solvent was removed, and the residue was
chromatographed on HP-20 (H.sub.2O-MeOH, 100:0 to 0:100) to give
320 (140 mg, 100%) as a white powder. .sup.1H-NMR (CD.sub.3OD)
.delta. 2.29 (s, 3H), 2.54 and 2.56 (2 s, total 3H), 2.82 (m, 2H),
3.48 (m, 2H), 3.65-5.80 (m, 9H), 4.09 and 4.21 (2 m, total 2H),
6.80 (m, 4H), 7.00 (t, J=7.5 Hz, 1H), 7.18 (m, 2H), 7.57 (d, J=7.8
Hz, 1H), 7.88 (m, 2H), 7.99 (m, 1H); MS (FAB), m/z 565 (M.sup.++1);
Anal. Calcd for C.sub.30H.sub.35N.sub.4O.sub.7Na.1.0H.sub.2O: C,
59.59; H, 6.17; N, 9.27. Found: C, 59.10; H, 6.28; N, 8.86.
Example 271
4-[2-N--(N'-methoxy-N'-methylamino)ethyl-N-[4-[N'-(2-bromophenyl)ureido]-3-
-methoxyphenyl acetamido]ethoxy]benzoic acid
[1484] ##STR1943##
[1485] To a stirred solution of methyl
4-[2-N-formylmethyl-N-[3-methoxy-4-[N'-(2-bromophenyl)ureido]phenyl]aceta-
mido]ethoxybenzoate (209 mg, 0.35 mmol) in EtOH (7 ml), was added
N-methoxy-N-methylamine HCl salt (341 mg, 3.5 mmol) at rt The
reaction was sonicated for 5 min. at rt, then cooled to 0.degree.
C. To the cooled solution, was added NaBH(OAc).sub.3 (370 mg, 1.75
mmol) and the stirring was continued for 18 h at rt. The mixture
was poured into sat. NaHCO.sub.3, extracted with CHCl.sub.3 (50
ml.times.3), and dried over MgSO.sub.4. After removal of the
solvent, the residue was chromatographed on TLC (Whatman, PLK-5F, 2
plates, CHCl.sub.3-MeOH, 98:2) to give methyl
4-[2-N--(N'-methoxy-N'-methylamino)ethyl-N-[4-[N'-(2-bromophenyl)ureido]--
3-methoxyphenyl acetamido]ethoxy]benzoate (89 mg, 40%) as a white
amorphous foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.52 and
2.60 (2 s, 3H), 2.80 (m, 2H), 3.48 and 3.50 (2 s, total 3H), 3.65
(m, 2H), 3.72 (s, 3H), 3.77 (m, 4H), 3.90 (s, 3H), 4.08 (m, 1H),
4.22 (m, 1H), 6.82 (m, 5H), 7.12 (s, 2H), 7.30 (m, 1H), 7.52 (d,
J=8.1 Hz, 1H), 7.94 (m, 3H), 8.15 (d, J=8.3 Hz, 1H); MS (ESI) m/z
643 (M.sup.+).
[1486] To a solution of methyl
4-[2-N--(N'-methoxy-N'-methylamino)ethyl-N-[4-[N'-(2-bromophenyl)ureido]--
3-methoxyphenylacetamido]ethoxy]benzoate (89 mg, 0.14 mmol) in
THF-MeOH (5:1, v/v, 6 ml), was added 0.5 N NaOH (1.4 ml, 0.7 mmol)
at rt, and heated to reflux in a glass sealed bottle. The stirring
was continued for 3 hours at reflux. The reaction was poured into
water, and was acidified with 1 N HCl to pH 5, extracted with
CHCl.sub.3-MeOH (2:1, v/v, 30 mL.times.3), dried over MgSO.sub.4.
The solvent was removed in vacuo to give 321 (53 mg, 60%) as a
white powder. .sup.1H-NMR (CD.sub.3OD, 400 MHz) .delta. 2.62 and
2.64 (2 s, total 3H), 2.80 (m, 2H), 3.50 (d, J=7.3 Hz, 3H),
3.63-3.88 (m, 6H), 4.12 (m, 1H), 4.25 (m, 1H), 6.82-7.00 (m, 5H),
7.30 (m, 1H), 7.58 (m, 1H), 7.95 (m, 4H); MS (FAB), m/z 629
(M.sup.+); Anal. Calcd for
C.sub.29H.sub.33BrN.sub.4O.sub.7.0.25H.sub.2O: C, 54.94; H, 5.33;
N, 8.84. Found: C, 55.39; H, 5.53; N, 8.23.
Example 272
4-[2-N--(N',N'-diallyl)ethyl-N-[3-methoxy-4-[N''-(2-methylphenyl)ureido]ph-
enyl]acetamido]ethoxybenzoic acid sodium salt
[1487] ##STR1944##
[1488] To a stirred solution of methyl
4-[2-N-formylmethyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acet-
amido]ethoxybenzoate (400 mg, 0.75 mmol) in EtOH (15 ml), were
added diallylamine (926 ml, 7.5 mmol) and acetic acid (429 ml, 7.5
mmol) at rt. The reaction was stirred for 5 min. at rt, then cooled
to 0.degree. C. To the cooled solution, was added NaBH.sub.3CN (118
mg, 1.9 mmol) and the stirring was continued for 1 h at rt. The
mixture was poured into sat. NaHCO.sub.3 and was extracted with
EtOAc (50 ml.times.3), washed with brine, and dried over
MgSO.sub.4. After removal of the solvent in vacuo, the residue was
chromatographed on silica gel (middle pressure chromatography
system: YAMAZEN YFLC-5404-FC, linear gradient toluene-acetone 9:1
to 1:1) to give methyl
4-[2-N--(N',N'-diallyl)ethyl-N-[3-methoxy-4-[N''-(2-methylphenyl)ureido]p-
henyl]acetamido]ethoxybenzoate (385 mg, 84%) as a white amorphous
foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.30 (s, 3H), 2.60
(m, 2H), 3.09 (m, 4H), 3.49 (m, 2H), 3.60 (s, 2H), 3.70 (m, 5H),
3.89 (s, 3H), 4.02 and 4.19 (2 m, total 2H), 5.01-5.20 (m, 4H),
4.79 (m, 2H), 6.30 and 6.32 (2 s, total 1H), 6.70-6.84 (m, 5H),
7.08 (m, 1H), 7.14 (m, 1H), 7.25 (m, 1H), 7.60 (m, 1H), 7.93 and
7.98 (2 d, J=8.8 Hz, 2H), 8.03 and 8.06 (2 d, J=8.3 Hz, 1H); MS
(FAB), m/z 615 (M.sup.++1).
[1489] To a stirred solution of methyl
4-[2-N--(N,N'-diallyl)ethyl-N-[3-methoxy-4-[N''-(2-methylphenyl)ureido]ph-
enyl]acetamido]ethoxybenzoate (385 mg, 0.63 mmol) in MeOH (3 ml),
was added 1N HCl (756 ml, 0.76 mmol) at rt. The reaction was
stirred for 5 min. at rt, then evaporated to give methyl
4-[2-N--(N',N'-diallyl)ethyl-N-[3-methoxy-4-[N''-(2-methylphenyl)ureido]p-
henyl]acetamido]ethoxybenzoate HCl salt (385 mg, 99%) as an
amorphous foam. Anal. Calcd for
C.sub.35H.sub.43ClN.sub.4O.sub.6.0.5H.sub.2O: C, 63.67; H, 6.72; N,
8.49. Found: C, 63.67; H, 6.69; N, 8.43.
[1490] To a solution of
4-[2-N--(N',N'-diallyl)ethyl-N-[3-methoxy-4-[N''-(2-methylphenyl)ureido]p-
henyl]acetamido]ethoxybenzoate (175 mg, 0.29 mmol) in THF-MeOH
(1:1, v/v, 20 ml), was added 0.25 N. NaOH (2.5 ml, 0.63 mmol) at
rt, and heated to reflux. The stirring was continued for 1 hours at
reflux. The solvent was removed, and the residue was
chromatographed on HP-20 (H.sub.2O-MeOH, 100:0 to 0:100) to give
322 (160 mg, 94%) as a white powder. .sup.1H-NMR (CD.sub.3OD)
.delta. 2.29 (s, 3H), 2.60 (t, J=6.9 Hz, 1H), 2.67 (t, J=7.0 Hz,
1H), 3.10 (d, J=6.6 Hz, 2H), 3.14 (d, J=6.6 Hz, 2H), 3.59 (m, 2H),
3.69-3.80 (m, 4H), 3.80 (s, 3H), 4.06 (t, J=5.2 Hz, 4.21 (t, J=5.1
Hz, 1H), 5.15 (m, 4H), 5.80 (m, 2H), 6.79 (m, 2H), 6.84 (d, J=8.8
Hz, 2H), 7.00 (t, J=7.5 Hz, 1H), 7.14 (m, 2 h), 7.48 (m, 1H), 7.91
(dd, J=6.1 Hz, 8.8 Hz, 2H), 8.00 (m, 1H); MS (FAB), m/z 601
(M.sup.+); Anal. Calcd for
C.sub.34H.sub.39N.sub.4O.sub.6Na.0.5H.sub.2O: C, 64.65; H, 6.38; N,
8.87. Found: C, 64.53; H, 6.58; N, 8.78.
Example 273
4-[2-N--(N',N'-diallyl)ethyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyphe-
nylacetamido]ethoxy)benzoic acid sodium salt
[1491] ##STR1945##
[1492] To a stirred solution of methyl
4-[2-N-formylmethyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy
phenylacetamido]ethoxy]benzoate (100 mg, 0.18 mmol) in EtOH (3.6
ml), was added diallylamine (223 ml, 1.81 mmol) at rt. The reaction
was stirred for 5 min. at rt, then cooled to 0.degree. C. To the
cooled solution, were added AcOH (104 ml, 1.81 mmol) and
NaBH.sub.3CN (28 mg, 0.45 mmol), and the stirring was continued for
18 h at rt. The mixture was poured into sat. NaHCO.sub.3, was
extracted with CHCl.sub.3 (30 mL.times.3), washed with brine, and
dried over MgSO.sub.4. After removal of the solvent in vacuo, the
residue was chromatographed on silica gel (middle pressure
chromatography system: YAMAZEN YFLC-5404-FC, linear gradient
CHCl.sub.3-MeOH 10:0 to 20:1) to give methyl
4-[2-N--(N',N'-diallyl)ethyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxyph-
enyl acetamido]ethoxy]benzoate (96 mg, 83%) as a white amorphous
foam. .sup.1H-NMR (CDCl.sub.3, 400 MHz) .delta. 2.60 (m, 2H), 3.09
(d, J=6.4 Hz, 1H), 3.11 (d, J=6.4 Hz, 3H), 3.52 (m, 2H), 3.61 (s,
2H), 3.70-3.80 (m, 5H), 3.86 (s, 3H), 4.05 and 4.20 (2 m, total
2H), 5.06-5.21 (m, 4H), 5.80 (m, 2H), 6.71-6.85 (m, 4H), 6.98 (m,
1H), 7.22 (m, 1H), 7.32 (m, 3H), 7.93 (d, J=7.8 Hz, 2H), 7.98 (m,
1H), 8.18 (dd, J=1.5 Hz, 8.2 Hz, 1H); MS (ESI) m/z 635
(M.sup.+).
[1493] To a solution of methyl
4-[2-N--(N',N'-diallyl)ethyl-N-[4-[N'-(2-chlorophenyl)ureido]-3-methoxy
phenylacetamido]ethoxy]benzoate (96 mg, 0.15 mmol) in THF-MeOH
(1:1, v/v, 8 ml), was added 0.25 N NaOH (3.91 ml, 0.98 mmol) at rt,
and heated to 50.degree. C. The stirring was continued for 6 hours
at reflux. The solvent was removed, and the residue was
chromatographed on HP-20 (H.sub.2O-MeOH, 0:100 to 100:0) to give
323 (88 mg, 94%) as a white powder. .sup.1H-NMR (CD.sub.3OD, 400
MHz) .delta. 2.61 (m, 2H), 3.10 (s, 2H), 3.12 (s, 2H), 3.59 (m,
2H), 3.70 (s, 2H), 3.78 (m, 2H), 3.82 (s, 3H), 4.10 (m, 1H), 4.21
(m, 1H), 5.18 (m, 4H), 5.81 (m, 2H), 6.82 (m, 4H), 7.01 (t, J=7.8
Hz, 1H), 7.25 (m, 1H), 7.39 (d, J=7.8 Hz, 1H), 7.90 (m, 2H), 8.02
(m, 2H); MS (ESI) m/z 621 (M.sup.+); Anal. Calcd for
C.sub.33H.sub.36ClN.sub.4O.sub.6.Na.1.25H.sub.2O: C, 59.55; H,
5.83; N, 8.42. Found: C, 59.90; H, 5.74; N, 7.96.
Example 274
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylacetoamido]-
ethyl-1-piperazinylacetic acid
[1494] ##STR1946##
[1495] To a stirred suspension of 1-(2-hydroxyethyl)piperazine
(5.21 g, 40.0 mmol) and K.sub.2CO.sub.3 (8.76 g, 63.4 mmol) in
CH.sub.3CN (100 ml) was added ethyl bromoacetate (5.60 ml, 50.5
mmol) at 0.degree. C. The reaction mixture was heated under reflux
for 5 h, diluted with EtOAc, and washed with water and brine. The
extract dried over Na.sub.2SO.sub.4, concentrated to dryness and
afforded ethyl 4-(2-hydroxyethyl)-1-piperazinylacetate (9.65 g,
100%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.23 (t,
3H, J=7.3 Hz), 2.51-2.61 (m, 1H), 3.22 (s, 2H), 3.61 (t, 2H, J=5.4
Hz), 4.19 (q, 2H, J=7.3 Hz).
[1496] To a solution of 2,4-dinitrobenzenesulfonyl chloride (1.0 g,
3.75 mmol) and pyridine (0.34 ml, 4.20 mmol) in THF (19 ml) was
added dropwise methylamine (2.0M THF solution, 2.3 ml, 4.60 mmol)
at 0.degree. C. The reaction mixture was stirred for 1 hr, quenched
by the addition of 1N HCl solution, and extracted with EtOAc. The
extract was washed with sat. NaHCO.sub.3 solution and brine, dried
over Na.sub.2SO.sub.4, and concentrated to dryness. The residue was
recrystallized from EtOAc-Et.sub.2O to give methyl
2,4-nitrobenzenesulfonamide (546 mg, 56%) as a colorless solid.
.sup.1H-NMR (DMSO) .delta. 2.60 (d, 3H, J=4.9 Hz), 8.22 (d, 1H,
J=8.8 Hz), 8.31 (q, 1H, J=4.9 Hz), 8.66 (dd, 1H, J=8.8, 2.0 Hz),
8.90 (d, 1H, J=2.0 Hz).
[1497] To a solution of ethyl
4-(2-hydroxyethyl)-1-piperazinylacetate (452 mg, 2.09 mmol), methyl
2,4-dinitrobenzenesulfonamide (546 mg, 2.09 mmol) and PPh.sub.3
(658 mg, 2.5 mmol) in THF was added DIAD (0.50 ml, 251 mmol) at
0.degree. C. After stirring 17 h at room temperature, the reaction
mixture was concentrated to dryness. Chromatography of the residue
with EtOAc-MeOH (10:1) to afford ethyl
4-[2-[N-(2,4-dinitrobenzensulfonyl)-N-methylamino]ethyl]-1-piperazinylace-
tate (864 mg, 90%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.27 (t, 3H, J=6.8 Hz), 2.35-2.63 (m, 10H), 2.98 (s, 3H),
3.20 (s, 2H), 3.41 (t, 2H, J=6.8 Hz), 4.17 (q, 2H, J=6.8 Hz), 8.33
(d, 1H, J=8.3 Hz), 8.46 (d, 1H, J=2.0 Hz), 8.50 (dd, 1H, J=8.3, 2.0
Hz).
[1498] A solution of ethyl
4-[2-[N-(2,4-dinitrobenzensulfonyl)-N-methylamino]ethyl]-1-piperazinylace-
tate (864 mg, 1.88 mmol), mercaptoacetic acid (0.17 ml, 2.44 mmol)
and Et.sub.3N (0.53 ml, 3.76 mmol) in CH.sub.2Cl.sub.2 (25 ml) was
stirred at rt for 3 hr. The reaction mixture ethyl
4-(2-methylaminoethyl)-1-piperazinylacetate (388 mg, 90%) as
reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.27 (t, 3H, J=6.8
Hz), 2.50 (s, 3H), 2.53-2.60 (m, 8H), 2.75 (t, 2H, J=5.9 Hz), 3.20
(s, 2H), 4.18 (q, 2H, J=6.8 Hz).
[1499] To a solution of ethyl
4-(2-methylaminoethyl)-1-piperazinylacetate (388 mg, 1.69 mmol),
Et.sub.3N (0.32 ml, 2.25 mmol) and DMAP (46 mg, 0.38 mmol) in DMF
(15 ml) was stirred for 15 min at room temperature, then (532 mg,
1.69 mmol), HOBt (103 mg, 0.76 mmol) and EDC.HCl (486 mg, 2.53
mmol) was added to the reaction mixture which was stirred for 15 h
at room temperature The reaction mixture was diluted with EtOAc,
which was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated to dryness. Chromatography of the residue with
CHCl.sub.3-MeOH (10:1, v/v) to afford ethyl
4-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylacetoamido-
]ethyl-1-piperazinylacetate (889 mg, mixture of DMF) as a reddish
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25-1.29 (m, 3H), 2.29 (s,
3H), 2.42-2.63 (m, 10H), 3.20, 3.18 (each s, total 3H), 3.55, 3.40
(each t, total 2H, J=6.8 Hz), 3.65, 3.69 (each s, total 2H), 3.72
(s, 3H), 4.15-4.21 (m, 2H), 6.50 (m, 1H), 6.77-6.81 (m, 8H),
7.11-7.24 (m, 3H), 7.53 (d, 1H, J=8.3 Hz), 8.02 (s, 1H), 8.06 (d,
1H, J=7.8 Hz).
[1500] To a stirred solution of ethyl
44-[2-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylacetoamid-
o]ethyl-1-piperazinylacetate (889 mg, 1.69 mmol) in THF-EtOH (5:1,
v/v, 18 ml) was added 4N NaOH (0.84 ml, 3.38 mmol). The reaction
mixture was stirred at rt for 4 h, adjusted to pH 7.5 with 1N HCl
and extracted with CHCl.sub.3-MeOH (4:1, v/v). The combined
extracts were dried over MgSO.sub.4 and concentrated to afforded
324 (218 mg, 26% 2 steps) as a brown amorphous foam. IR (KBr) n
3299, 3004, 1700, 1627, 1598, 1536 cm.sup.-1; .sup.1H-NMR (DMSO)
.delta. 2.25 (s, 3H), 2.36-2.62 (m, 10H), 2.84, 2.99 (each s, total
3H), 3.13, 3.14 (each s, total 2H), 3.38-3.45 (m, 2H), 3.61, 3.65
(each s, total 2H), 3.86 (s, 3H), 6.74 (t, 1H, J=7.8 Hz), 6.87 (s,
1H), 6.93 (t, 1H, J=7.8 Hz), 7.11-7.17 (m, 2H), 7.79 (d, 1H, J=7.8
Hz), 8.01 (d, 1H, J=7.8 Hz), 8.47 (s, 1H), 8.57 (s, 1H); MS (FAB)
m/z 498 (M.sup.++1); Anal. Calcd for
C.sub.26H.sub.35NO.sub.5.2HCl.H.sub.2O: C, 53.06; H, 6.67; N,
11.89. Found: C, 53.04; H, 6.15; N, 11.09.
Example 275
1-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]-
ethyl]-4-piperidinylacetic acid
[1501] ##STR1947##
[1502] To a stirred solution of
2-(N-benzyloxycarbonyl-N-methylamino)acetaldehyde (2.07 g, 10.0
mmol) and ethyl 4-piperidinylideneacetate (1.69 g, 10.0 mmol) in
MeOH--AcOH (10:1, v/v, 22 ml) was added NaBH.sub.3CN (1.32 g, 20
mmol) and the stirring was continued overnight. The mixture was
quenched by addition of sat. NaHCO.sub.3 (200 ml) and extracted
with CHCl.sub.3 (3.times.150 ml). The combined extracts were dried
over MgSO.sub.4 and evaporated. The residue was chromatographed on
silica-gel with CHCl.sub.3-EtOH (40:1, v/v) to give ethyl
1-[2-(N-benzyloxycarbonyl-N-methylamino)ethyl]-4-piperidinylideneacetate
(1.71 g, 47%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.25 (t, J=7.3 Hz, 3H), 2.16 (m, 2H), 2.57 (m, 4H), 2.95 (m, 7H),
3.44 (m, 2H), 4.13 (q, J=7.3 Hz, 2H), 5.12 (s, 2H), 5.49-5.53 (m,
1H), 7.35 (m, 5H).
[1503] A solution of ethyl
1-[2-(N-benzyloxycarbonyl-N-methylamino)ethyl]-4-piperidinylidene
acetate (1.70 g, 4.72 mmol) in EtOH--AcOH (20:1, v/v, 21 ml) was
hydrogenated over 5% Pd/C (2 g) for 3 days with stirring. The
mixture was filtered and the filtrate was concentrated in vacuo.
The residue was made basic with sat. NaHCO.sub.3 and extracted with
CHCl.sub.3 (300 ml). The extract was dried over Na.sub.2CO.sub.3
and evaporated to give ethyl
1-(2-methylaminoethyl)-4-piperidinylacetate (813 mg, 75%) as a
yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25 (t, J=7.3 Hz,
3H), 1.68-1.81 (m, 5H), 1.97 (t, J=11.2 Hz, 2H), 2.22 (d, J=7.3 Hz,
2H), 2.43-2.47 (m, 5H), 2.66 (t, J=6.4 Hz, 2H), 2.85-2.90 (m, 2H),
4.13 (q, J=7.3 Hz, 2H).
[1504] To a stirred solution of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (550 mg,
1.75 mmol) and ethyl 1-(2-methylaminoethyl)-4-piperidinylacetate
(400 mg, 1.75 mmol) in DMF (10 ml) were added EDC.HCl (503 mg, 2.63
mmol), HOBt (cat.), and DMAP (cat.) and the stirring was continued
overnight. The mixture was diluted with EtOAc (300 ml), washed with
brine (200 ml), dried over MgSO.sub.4, and evaporated. The residue
was chromatographed on silica gel with CHCl.sub.3-EtOH (10:1, v/v)
to give ethyl
1-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido-
]ethyl]-4-piperidinylacetate (697 mg, 76%) as a yellow gum.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.19-2.06 (series of m, 12H), 2.21
(t, J=7.8 Hz, 2H), 2.28 (s, 3H), 2.41 (t, J=7.3 Hz, 1H), 2.46 (t,
J=7.3 Hz, 1H), 2.80-2.89 (m, 2H), 2.95 and 3.01 (s, each, total
3H), 3.40 and 3.50 (t, J=6.8 Hz, each, total 2H), 3.64-3.75 (m,
5H), 4.09-4.16 (m, 2H), 6.59 (s, 1H), 6.77-6.79 (m, 2H), 7.12 (t,
J=7.3 Hz, 1H), 7.21-7.27 (m, 3H), 7.54 (d, J=8.3 Hz, 1H), 8.06 (dd,
J=8.3, 2.4 Hz, 1H).
[1505] To a stirred solution of ethyl
1-[2-[N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]acetamido-
]ethyl]-4-piperidinylacetate (690 mg, 1.32 mmol) in THF (11 ml) was
added 0.25 N aq. NaOH (11 ml, 2.75 mmol) and the stirring was
continued overnight. The mixture was diluted with H.sub.2O (50 ml),
neutralized with 1 N HCl, and extracted with CHCl.sub.3-MeOH (2:1,
v/v, 3.times.100 ml). The combined extracts were dried over
MgSO.sub.4 and evaporated. The residue was dissolved in MeOH (50
ml) and activated carbon (2 g) was added to this solution. The
suspension was refluxed for 30 min with stirring and filtered
through Celite. The filtrate was evaporated and the residue was
triturated by taking up CHCl.sub.3 and adding hexane until a
precipitate formed. This precipitate was collected and dried in
vacuo to give 325 (75 mg, 11%) as a white amorphous solid.
.sup.1H-NMR (DMSO) .delta. 1.19-2.99 (series of m, total 17H),
3.32-3.43 (m, 4H), 3.62-3.65 (m, 2H), 3.86 (s, 3H), 6.73 (t, J=8.3
Hz, 1H), 6.87 (s, 1H), 6.93 (t, J=7.8 Hz, 1H), 7.11-7.17 (m, 2H),
7.79 (d, J=8.3 Hz, 1H), 8.01 (d, J=8.3 Hz, 1H), 8.47 (s, 1H), 8.57
(s, 1H); MS-FAB m/z 497 (M.sup.++1); Anal. Calcd for
C.sub.27H.sub.36N.sub.4O.sub.5.HCl: C, 60.84; H, 7.00; N, 10.51.
Found: C, 60.97; H, 7.14; N, 10.17.
Example 276
1-[2-[N-methyl-N-[4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]ethyl]-4-p-
iperidinylacetic acid
[1506] ##STR1948##
[1507] To a stirred solution of ethyl
1-(2-methylaminoethyl)-4-piperidinylacetate (400 mg, 1.75 mmol) and
Et.sub.3N (366 ul, 2.63 mmol) in DMF (10 ml) was added
pentafluorophenyl 4-(N'-(2-methylphenyl)ureido)phenylacetate (788
mg, 1.75 mmol) and the stirring was continued overnight. The
mixture was diluted with EtOAc (300 ml), washed with brine (200
ml), dried over MgSO.sub.4, and evaporated. The residue was
chromatographed on silica-gel with CHCl.sub.3-EtOH (10:1, v/v) to
give ethyl
1-[2-[N-methyl-N-[4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]ethyl]-4--
piperidinyl acetate (630 mg, 73%) as a colorless oil.
[1508] To a stirred solution of ethyl
1-[2-[N-methyl-N-[4-[N'-(2-methylphenyl)ureido]phenyl]acetamido]ethyl]-4--
piperidinylacetate (630 mg, 1.27 mmol) in THF (10 ml) was added
0.25 N aq. NaOH (10 ml) and the stirring was continued overnight.
The reaction mixture was diluted with H.sub.2O (100 ml),
neutralized with 1 N HCl, and extracted with CHCl.sub.3-MeOH (2:1,
v/v, 3.times.100 ml). The combined extracts were dried over
MgSO.sub.4 and evaporated. The residue was triturated by taking up
CHCl.sub.3 and adding hexane until precipitate formed. This
precipitate was collected and dried in vacuo to give 326 (20 mg,
3%) as a white amorphous solid. .sup.1H-NMR (DMSO) .delta. 1.69 (m,
5H), 2.15 (m, 4H), 2.24 (s, 2H), 2.50 (m, 2H), 2.83 and 2.99 (s,
each, total 3H), 3.32-3.49 (m, 4H), 3.63 (d, J=6.8 Hz, 2H),
6.91-6.95 (m, 1H), 7.13 (m, 4H), 7.39 (d, J=8.3 Hz, 2H), 7.83 (d,
J=7.3 Hz, 1H), 7.97 (m, 1H), 9.12 (m, 1H); MS (FAB): m/z 467
(M.sup.++1).
Example 277
4-[2-N-[4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylacetamido]ethyl-1-pip-
erazinylacetic acid
[1509] ##STR1949##
[1510] To a solution of ethyl
4-(2-methylaminoethyl)-1-piperazinylacetate (700 mg, 3.05 mmol),
Et.sub.3N (0.64 ml, 4.58 mmol) and DMAP (75 mg, 0.61 mmol) in THF
(15 ml) was stirred for 30 min at at, then
4-[N'-(2-methylphenyl)ureido]phenylacetic acid (917 mg, 3.05 mmol),
HOBt (82 mg, 0.61 mmol) and EDC.HCl (879 mg, 4.58 mmol) was added
to the reaction mixture which was stirred for 12 h at rt. The
reaction mixture was diluted with EtOAc, which was washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated to dryness.
Chromatography of the residue with CHCl.sub.3-MeOH (10:1, v/v)
afforded ethyl
4-[2-N-[4-[N'-(2-methylphenyl)ureido]phenyl]-N-methylacetamido]ethyl-1-pi-
perazinylacetate (996 mg, 66%) as a yellow amorphous foam.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.25-1.29 (m, 3H), 2.20 (s, 3H),
2.47-2.58 (m, 10H), 2.97, 3.05 (each s, total 3H), 3.17, 3.20 (each
s, total 2H), 3.45, 3.52 (each d, total 2H, J=6.8 Hz), 3.64, 3.68
(each s, 2H), 4.15-4.21 (m, 2H), 7.01-7.19 (m, 8H), 7.48 (m, 1H),
7.64 (m, 1H); MS (FAB) m/z 496 (M.sup.++1).
[1511] To a stirred solution of ethyl
4-[2-N-[4-[N'-(2-methylphenyl)ureido]phenyl]-N-methyl
acetamido]ethyl-1-piperazinylacetate (996 mg, 2.01 mmol) in
THF-EtOH (5:1, 12 ml) was added 4N NaOH (1.0 ml, 4.00 mmol). The
reaction mixture was stirred at rt for 4 h, adjusted to pH 7.5 with
1N HCl and extracted with CHCl.sub.3-MeOH (4:1, v/v). The combined
extracts were dried over MgSO.sub.4 and concentrated to afforded
327 (73 mg, 8%) as a yellow amorphous foam. IR (KBr) n 3338, 2925,
2850, 2821, 1704, 1627, 1540 cm.sup.-1; .sup.1H-NMR (DMSO) .delta.
2.24 (s, 3H), 2.33-2.61 (m, 10H), 2.82, 2.95 (each s, total 3H),
3.00, 3.02 (each s, total 2H), 3.39 (t, 2H, J=6.8 Hz), 3.60, 3.62
(each s, total 2H), 6.92 (t, 1H, J=7.8 Hz), 7.09-7.16 (m, 4H),
7.41-7.44 (m, 2H), 7.76, 7.77 (each d, 2H, J=7.8 Hz), 8.46, 8.53
(each s, 1H), 9.54, 9.59 (each s, 1H); MS (FAB) m/z 468
(M.sup.++1); Anal. Calcd for C.sub.25H.sub.33N.sub.5O.sub.5.2.HCl:
C, 55.56; H, 6.53; N, 12.96. Found: C, 54.99; H, 6.45; N,
11.58.
Example 278
4-[2-N-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenyl]-N-methylacetamido]e-
thyl-1-piperazinylacetic acid
[1512] ##STR1950##
[1513] To a solution of ethyl
4-(2-methylaminoethyl)-1-piperazinylacetate (695 mg, 3.03 mmol),
Et.sub.3N (0.64 ml, 4.58 mmol) and DMAP (75 mg, 0.61 mmol) in DMF
(15 ml) was stirred for 15 min at rt, then
4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenylacetic acid (965 mg,
3.03 mmol), HOBt (82 mg, 0.61 mmol) and EDC.HCl (872 mg, 4.54 mmol)
was added to the reaction mixture which was stirred for 12 h at rt.
The reaction mixture was diluted with EtOAc, which was washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated to dryness.
Chromatography of the residue with CHCl.sub.3-MeOH (10:1, v/v)
afforded ethyl
4-[2-N-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenyl]-N-methyl
acetamido]ethyl-1-piperazinylacetate (1.21 g, mixture of DMF) as a
black oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.24-1.29 (m, 3H),
2.45-2.59 (m, 10H), 2.98, 3.05 (each s, total 3H), 3.17, 3.20 (each
s, total 2H), 3.44, 3.52 (each t, total 2H, J=6.8 Hz), 3.66 (s,
2H), 4.15-4.21 (m, 2H), 6.77-6.78 (m, 2H), 6.79-7.11 (m, 3H),
7.68-7.95 (m, 2H), 7.64 (broad s, 1H), 8.20 (t, 1H, J=7.8 Hz); MS
(FAB) m/z 530 (M.sup.++1).
[1514] To a stirred solution of ethyl
4-[2-N-[4-[N'-(2-fluorophenyl)ureido]-3-methoxyphenyl]-N-methyl
acetamido]ethyl-1-piperazinylacetate (1.21 g, mixture of DMF) in
THF-EtOH (5:1, v/v, 12 ml) was added 4N NaOH (1.0 ml, 4.00 mmol).
The reaction mixture was stirred at rt for 4 h, adjusted to pH 7.5
with 1N HCl and extracted with CHCl.sub.3-MeOH (4:1, v/v). The
combined extracts were dried over MgSO.sub.4 and concentrated to
afforded 328 (78 mg, 5% 2 steps) as a brown amorphous foam. IR
(KBr) n 3299, 2940, 2830, 1704, 1627, 1598, 1536 cm.sup.-1;
.sup.1H-NMR (DMSO) .delta. 2.36-2.61 (m, 10H), 2.83, 2.98 (each s,
total 3H), 3.11 (s, 2H), 3.37-3.43 (m, 2H), 3.62, 3.65 (each s,
total 2H), 3.85 (s, 3H), 6.75 (m, 1H), 6.87 (s, 1H), 6.98 (s, 1H),
7.12 (t, 1H, J=7.8 Hz), 7.20, 7.23 (each d, 2H, J=7.8 Hz), 8.01 (d,
1H, J=7.8 Hz), 8.17 (t, 1H, J=7.8 Hz), 8.72 (s, 1H), 9.19 (s, 1H);
MS (FAB) m/z 502 (M.sup.++1); Anal. Calcd for
C.sub.25H.sub.32FN.sub.5O.sub.5.2HCl.0.5H.sub.2O: C, 51.46; H,
6.05; N, 12.00. Found: C, 51.08; H, 5.69; N, 11.27.
Example 279
3-fluoro-1-[2-N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]ac-
etamido]ethyl-4-piperidinylacetic acid
[1515] ##STR1951##
[1516] To a stirred solution of 1-tert-butoxycarbonyl-4-piperidone
(14.9 g, 74.8 mmol) in DMF (35 mL) was added TMSCl (11.4 mL, 89.7
mmol) and then Et.sub.3N (25.0 mL, 179 mmol) dropwise at room
temperature, and the reaction mixture was heated at 80.degree. C.
for 18 hr. Hexane was added to the reaction mixture, and the
resulting mixture was washed with sat. NaHCO.sub.3 and brine, dried
over Na.sub.2SO.sub.4, and concentrated to dryness. Chromatography
of the residue with hexane-EtOAc (5:1, v/v) as eluent gave
1-tert-butoxycarbonyl-1,2,3,6-tetrahydro-4-(trimethylsilyloxy)pyridine
(20.4 g, 99%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
0.19 (s, 9H), 1.46 (s, 9H), 2.05-2.15 (m, 2H), 3.48-3.56 (m, 2H),
3.83-3.91 (m, 2H), 4.79 (broad s, 1H).
[1517] To a solution of
1-tert-butoxycarbonyl-1,2,3,6-tetrahydro-4-trimethylsilyloxy)pyridine
(20.4 g, 75.0 mmol) in CH.sub.3CN (500 mL) was added
Selectfluor.TM. (29.2 g, 82.5 mmol) at room temperature, and the
reaction mixture was stirred for 2 hr. EtOAc was added to the
reaction mixture, and the mixture was washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to dryness. Chromatography of
the residue with CHCl.sub.3-MeOH (6:1, v/v) as eluent gave
1-tert-butoxycarbonyl-3-fluoro-4-piperidone (14.5 g, 89%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.50 (s, 9H), 2.44
(t, J=6.9 Hz, 1H), 2.48-2.63 (m, 2H), 3.26 (ddd, J=13.5, 10.5, 3.9
Hz, 1H), 3.72 (t, J=6.9 Hz, 1H), 4.16 (m, 1H), 4.42 (m, 1H), 4.83
(dt, 49.2, 6.9 Hz, 1H).
[1518] To a solution of triethyl phosphonoacetate (3.72 g, 16.6
mmol) in THF (70 mL) was added lithium bis(trimethylsilyl)amide
(1.0M THF solution, 15.5 mL, 15.5 mmol) at -78.degree. C. After
being stirred at the same temperature for 1 hr,
1-tert-butoxycarbonyl-3-fluoro-4-piperidone (3.02 g, 13.9 mmol) was
added to the reaction mixture. The mixture was stirred for 30 min
at the same temperature, quenched by the addition of sat.
NH.sub.4Cl solution and extracted with EtOAc. The extracts were
washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to
dryness. Chromatography of the residue with hexane-EtOAc (8:1, v/v)
as eluent gave ethyl
(1-butoxycarbonyl-3-fluoropiperidin-4-yliden)acetate (3.23 g, 81%)
as a colorless solid. .sup.1H-NMR .delta. 1.30 (t, J=7.1 Hz, 3H),
1.48 (s, 9H), 2.10 (m, 1H), 2.56 (m, 1H), 2.77 (m, 1H), 3.13-3.54
(m, 2H), 3.70 (m, 1H), 4.17 4.18 (each q, J=7.1 Hz, total 2H), 5.82
5.98 (eachs, total 1H), 6.41 (each d, J=46.9 Hz, total 1H); MS
(FAB) m/z 288 (M.sup.++1).
[1519] A solution of ethyl
(1-butoxycarbonyl-3-fluoropiperidin-4-yliden)acetate (1.32 g, 4.59
mmol) in THF (30 mL) was hydrogenated over Pd--C (TMEDA complex,
66.0 mg) at room temperature for 2 hr under hydrogen atmosphere.
The catalyst was filtered off and the filtrate was concentrated to
dryness. Chromatography of the residue with hexane-EtOAc (9:1, v/v)
as eluent gave ethyl
1-tert-butoxycarbonyl-3-fluoro-4-piperidinylacetate (653 mg, 73%)
as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.26 (t, J=7.1
Hz, 3H), 1.45 (s, 9H), 1.53-1.79 (m, 2H), 1.92-2.09 (m, 2H), 2.31
(dd, J=16.4, 6.9 Hz, 1H), 2.52 (dd, J=16.4, 7.3 Hz, 1H), 2.61-3.06
(m, 2H), 4.14 (q, J=7.1 Hz, 2H), 4.28-4.77 (m, 2H); .sup.13C NMR
(CDCl.sub.3) 14.29, 25.80, 28.42, 35.99, 36.20, 60.55, 79.78,
86.72, 88.48, 154.94, 171.93; FAB-MS m/z 290 (M.sup.++1).
[1520] To a solution of ethyl
1-tert-butoxycarbonyl-3-fluoro-4-piperidinylacetate (653 mg, 2.26
mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA (5 mL) at 0.degree.
C. After being stirred at room temperature for 4 hr, the reaction
mixture was concentrated. The residue was taken up with sat
NaHCO.sub.3 solution and extracted with THF-EtOAc (1:1, v/v). The
extracts were dried over MgSO.sub.4 and concentrated to afford
ethyl 3-fluoro-4-piperidinylacetate (420 mg, 98%) as a yellow oil.
.sup.1H-NMR (CDCL.sub.3) .delta. 1.26 (t, J=7.4 Hz, 3H), 1.68-1.80
(m, 2H), 2.18 (m, 1H), 2.32 (dd, J=16.6, 6.8 Hz, 1H), 2.54 (dd,
J=16.6, 7.5 Hz, 1H), 2.82 (m, 1H), 2.90, 3.00 (each d, J=14.4 Hz,
total 1H), 3.31 (m, 1H), 3.51 (m, 1H), 4.15 (q, J=7.4 Hz, 2H),
4.82, 4.71 (each broad s, total 1H).
[1521] To a solution of ethyl 3-fluoro-4-piperidinylacetate (230
mg, 1.22 mmol) and 2-(N-tert-butoxy
carbonyl-N-methylamino)acetaldehyde (211 mg, 1.22 mmol) in THF (5
mL) was added NaBH(OAc).sub.3 (386 mg, 1.82 mmol) and acetic acid
(70.0 mL, 1.22 mmol) at room temperature After being stirred for 24
hr, the reaction mixture was quenched by the addition of sat.
NaHCO.sub.3 solution and extracted with EtOAc. The extracts were
washed with brine, dried over Na.sub.2CO.sub.3, and concentrated to
dryness. Chromatography of the residue with CHCl.sub.3-MeOH (6:1,
v/v) as eluent gave ethyl
3-fluoro-1-[2-(N-tert-butoxycarbonyl-N-methylamino)ethyl]-4-piperidinylac-
etate (236 mg, 56%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3)
.delta. 1.25 (t, J=7.1 Hz, 3H), 1.45 (s, 9H), 1.53 1.79 (m, 2H),
1.90-2.09 (m, 2H), 2.15 (dd, J=16.4, 7.1 Hz, 1H), 2.45-2.58 (m,
2H), 2.87 (s, 3H), 2.90-2.99 (m, 2H), 3.20 (m, 1H), 3.24-3.45 (m,
2H), 4.14 (q, J=7.1 Hz, 2H), 4.61, 4.73 (each broad s, 1H); ESI-MS
m/z 347.
[1522] To a solution of ethyl
3-fluoro-1-[2-(N-tert-butoxycarbonyl-N-methylamino)ethyl]-4-piperidinylac-
etate (236 mg, 0.68 mmol) in CH.sub.2Cl.sub.2 (10 mL) was added TFA
(5 mL) at 0.degree. C. After being stirred at room temperature for
4 hr, the reaction mixture was concentrated. The residue was taken
up with sat. NaHCO.sub.3 solution and extracted with CHCl.sub.3.
The extracts were dried over MgSO.sub.4 and concentrated to afford
ethyl 3-fluoro-1-[2-(N-methylamino)ethyl]-4-piperidinylacetate (117
mg, 70%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.26
(t, J=7.1 Hz, 3H), 1.58 (m, 1H), 1.70 (m, 1H), 1.99 (m, 1H), 2.14
(m, 1H), 2.27-2.33 (m, 1H), 2.47 (s, 3H), 2.48-2.56 (m, 4H), 2.72
(t, J=6.3 Hz, 2H), 2.90 (m, 1H), 3.16 (m, 1H), 4.14 (q, J=7.1 Hz,
2H), 4.67 (d, J=48.3 Hz, 1H); ESI-MS m/z 247 (M.sup.++1).
[1523] To a solution of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (164 mg,
0.52 mmol), ethyl
3-fluoro-1-[2-(N-methylamino)ethyl]-4-piperidinylacetate (117 mg,
0.47 mmol), Et.sub.3N (0.10 mL, 0.71 mmol), and HOBt (13.0 mg, 0.09
mmol) in THF (5 mL) was added EDC.HCl (137 mg, 0.71 mmol). After
being stirred at room temperature for 8 hr, the reaction mixture
was diluted with water and extracted with EtOAc. The extracts were
washed with brine, dried over Na.sub.2SO.sub.4, and concentrated to
dryness. Chromatography of the residue with toluene-acetone (1:2,
v/v) as eluent gave ethyl
3-fluoro-1-[2-N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]a-
cetamido]ethyl-4-piperidinylacetate (160 mg, 62%) as a yellow
amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.24-1.28 (m,
3H), 1.51-2.23 (m 7H), 2.26 (s, 3H), 2.35 (s, 2H), 2.39-2.56 (m,
3H), 2.88 (m, 1H), 2.95, 3.03 (each s, total 3H), 3.11 (m, 1H),
3.44 (m, 1H), 3.56 (m, 1H), 3.64 (s, 2H), 3.68 (s, 3H), 4.11-4.17
(m, 2H), 4.57, 4.69 (each s, total 1H), 6.72-6.82 (m, 2H),
7.10-7.33 (m, 5H), 7.54 (d, J=8.1 Hz, 1H), 8.06 (d, J=8.1 Hz, 1H);
ESI-MS m/z 543 (M.sup.++1).
[1524] To a solution of ethyl
3-fluoro-1-[2-N-methyl-N-[3-methoxy-4-[N'-(2-methylphenyl)ureido]phenyl]a-
cetamido]ethyl-4-piperidinylacetate (160 mg, 0.29 mmol) in THF (3
mL) was added 0.25N NaOH (1.30 mL, 0.32 mmol). After being stirred
at room temperature for 1 hr, the reaction mixture was
concentrated. The residue was diluted with water and neutralized
with 1N HCl at 0.degree. C. The mixture was concentrated and
purified by ion-exchanged resin (HP-20, Mitsubishi Chemical) to
give 329 (110 mg, 74%) as a yellow amorphous solid. IR (KBr) 3343,
2937, 1700, 1617, 1589, 1535, 1486, 1455, 1417 cm.sup.-1;
.sup.1H-NMR (CD.sub.3OD) .delta. 1.57-1.72 (m, 2H), 1.98 (m, 1H),
2.26 (m, 1H), 2.28 (s, 3H), 2.37-2.59 (m, 3H), 2.66-2.89 (m, 2H),
2.95, 3.09 (each s, total 3H), 3.14 (m, 1H), 3.40 (m, 1H), 3.51 (m,
1H), 3.59 (m, 1H), 3.71 (s, 2H), 3.78 (m, 1H), 3.89 (s, 3H), 4.69,
4.81 (each s, total 1H), 6.79 (dd, J=8.1, 1.5 Hz, 1H), 6.90 (broad
s, 1H), 7.01 (t, J=7.8 Hz, 1H), 7.13-7.19 (m, 2H), 7.58 (d, J=7.8
Hz, 1H), 8.00 (d, J=8.1 Hz, 1H); ESI-MS m/z 515 (M.sup.++1); Anal.
Calcd for C.sub.27H.sub.35FN.sub.4O, H.sub.2O: C, 60.89; H, 7.00;
N, 10.52. Found: C, 61.09; H, 6.80; N, 9.87.
Example 280
4-[2-N-[2-(4-fluorophenoxy)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)ureid-
o]phenyl]acetamido]ethylpiperazinyl-1-acetic acid
[1525] ##STR1952##
[1526] To a solution of
2-(N-benzyl-N-tert-butoxycarbonylamino)ethanol (6.85 g, 27.3 mmol),
4-fluorophenol (3.07 g, 27.3 mmol) and PPh.sub.3 (7.83 g, 30.0
mmol) in THF (100 mL) was added DIAD (6.00 mL, 30.0 mmol) at room
temperature After being stirred for 3 hr, the reaction mixture was
concentrated. Chromatography of the residue with hexane-EtOAc (8:1,
v/v) as eluent gave
1-[2-(N-benzyl-N-tert-butoxycarbonylamino)ethoxy]-4-fluorobenzene
(8.19 g, 64%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.42-1.50 (m, 9H), 3.41-3.67 (m, 2H), 3.92-4.11 (m, 2H), 4.51-4.63
(m, 2H), 6.73-6.85 (m, 2H), 6.89-6.98 (m, 2H), 7.24 (m 5H); FAB-MS
m/z 346 (M.sup.++1).
[1527] To a solution of
1-[2-(N-benzyl-N-tert-butoxycarbonylamino)ethoxy]-4-fluorobenzene
(8.19 g, 23.7 mmol) in CH.sub.2Cl.sub.2 (50 mL) was added TFA (40
mL) at 0.degree. C. After being stirred at room temperature for 1
hr, the reaction mixture was concentrated. The residue was taken up
with sat. NaHCO.sub.3 solution and extracted with CHCl.sub.3. The
extracts were washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated to give 1-(2-N-benzylaminoethoxy)-4-fluorobenzene
(4.38 g, 75%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta.
3.00 (t, J=5.2 Hz, 2H), 3.87 (s, 2H), 4.04 (t, J=5.2 Hz, 2H), 6.80
6.85 (m, 2H), 6.92-6.98 (m, 2H), 7.23-7.36 (m, 5H); FAB-MS m/z 246
(M.sup.++1).
[1528] A mixture of 1-(2-N-benzylaminoethoxy)-4-fluorobenzene (1.08
g, 4.40 mmol), ethyl 4-(2-bromoethyl)piperazinyl-1-acetate (1.23 g,
4.40 mmol), and K.sub.2CO.sub.3 (0.61 g, 17.9 mmol) in CH.sub.3CN
(50 mL) was heated under reflux for 8 hr. The resulting mixture was
filtered and the filtrate was concentrated to dryness.
Chromatography of the residue with toluene-acetone (3:1, v/v) as
eluent gave ethyl
4-[2-N-benzyl-N-[2-(4-fluorophenoxy)ethyl]amino]ethylpiperazinyl-1-acetat-
e (1.51 g, 77%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.27 (t, J=7.1 Hz, 3H), 2.31-2.66 (m, 10H), 2.75 (t, J=6.6 Hz, 2H),
2.90 (t, J=6.1 Hz, 2H), 3.18 (s, 2H), 3.72 (s, 2H), 3.97 (t, J=6.1
Hz, 2H), 4.17 (q, J=7.1 Hz, 2H), 6.75-6.79 (m, 2H), 6.91-6.96 (m,
2H), 7.21-7.35 (m, 5H); FAB-MS m/z 444 (M.sup.++1).
[1529] A solution of ethyl
4-[2-N-benzyl-N-[2-(4-fluorophenoxy)ethyl]amino]ethylpiperazinyl-1-acetat-
e (1.50 g, 3.38 mmol) in EtOH (30 mL) was hydrogenated over 5%
Pd--C (53.1% wet, 0.73 g) under hydrogen atmosphere for 4 hr. The
catalyst was filtered off and the filtrate was concentrated. The
residue was taken up with sat. NaHCO.sub.3 solution and extracted
with CHCl.sub.3. The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to afford ethyl
4-[2-N-[2-(4-fluorophenoxy)ethyl]amino]ethylpiperazinyl-1-acetate
(1.12 g, 94%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.27 (t, J=7.1 Hz, 3H), 1.81 (broad s, 1H), 2.47-2.66 (m, 12H),
3.00 (t, J=5.4 Hz, 2H), 3.20 (s, 2H), 4.03 (t, J=5.4 Hz, 2H), 4.18
(q, J=7.1 Hz, 2H), 6.81-6.85 (m, 2H), 6.90-6.99 (m, 2H); FAB-MS m/z
354 (M.sup.++1).
[1530] To a solution of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (485 mg,
1.54 mmol), ethyl
4-[2-N-[2-(4-fluorophenoxy)ethyl]amino]ethylpiperazinyl-1-acetate
(545 mg, 1.54 mmol), Et.sub.3N (0.32 mL, 2.32 mmol), and HOBt (41.5
mg, 0.31 mmol) in THF (15 mL) was added EDC.HCl (883 mg, 2.32 mmol)
at room temperature After stirring for 24 hr, the reaction mixture
was diluted with water and extracted with CHCl.sub.3-MeOH (10:1,
v/v). The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to dryness. Chromatography of
the residue with CHCl.sub.3-MeOH (4:1, v/v) as eluent to give ethyl
4-[2-N-[2-(4-fluorophenoxy)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)urei-
do]phenyl]acetamido]ethylpiperazinyl-1-acetate (532 mg, 53%) as a
yellow amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.26, 1.27
(each t, J=7.1 Hz, total 3H), 2.27 (s, 3H), 2.51-2.63 (m, 10H),
3.16, 3.19 (each s, total 2H), 3.51-3.56 (m, 2H), 3.63, 3.67 (each
s, total 2H), 3.69-3.81 (m, 5H), 3.95, 4.10 (each t, J=5.2 Hz,
total 2H), 4.16, 4.18 (each q, J=7.1 Hz, total 2H), 6.56 (d, J=8.1
Hz, 1H), 6.72-6.78 (m, 4H), 6.89-6.98 (m, 2H), 7.12 (t, J=7.6 Hz,
1H), 7.20-7.23 (m, 3H), 7.51 (d, J=7.6 Hz, 1H), 8.04 (d, J=8.1 Hz,
1H); FAB-MS m/z 650 (M.sup.++1).
[1531] To a solution of ethyl
4-[2-N-[2-(4-fluorophenoxy)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)urei-
do]phenyl]acetamido]ethylpiperazinyl-1-acetate (532 mg, 0.82 mmol)
in dioxane (8 mL) was added dropwise 0.25N NaOH (5.00 mL, 1.25
mmol) at room temperature, and the reaction mixture was stirred for
1 hr. The resulting mixture was concentrated, diluted with water,
and neutralized with 1N HCl at 0.degree. C. The mixture was
extracted with CHCl.sub.3-MeOH (4:1, v/v). The extracts were washed
with brine, dried over Na.sub.2SO.sub.4, and concentrated to
dryness. Chromatography of the residue with CHCl.sub.3-MeOH (3:1,
v/v) as eluent gave 330 (168 mg, 33%) as a pale yellow amorphous
solid. IR (KBr) 3338, 2938, 2829, 1635, 1533, 1506, 1454, 1415
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25 (s, 3H),
2.37-2.48 (m, 6H), 2.53-2.67 (m, 6H), 3.09 (m, 2H), 3.44-3.49 (m,
2H), 3.64-3.69 (m, 2H), 3.72 (s, 2H), 3.80, 3.84 (each s, total
3H), 4.06-4.09 (m, 2H), 6.73 (m, 1H), 6.85 (s, 1H), 6.91-6.97 (m,
3H), 7.07-7.18 (m, 4H), 7.78 (d, J=8.1 Hz, 1H), 8.00 (dd, J=8.1,
2.7 Hz, 1H), 8.53 (m, 1H), 8.59 (m, 1H); FAB-MS m/z 622
(M.sup.++1); Anal. Calcd for C.sub.33H.sub.40FN.sub.5O.sub.6.2HCl:
C, 57.06; H, 6.09; N, 10.08. Found: C, 56.83; H, 6.05; N, 9.90.
Example 281
4-[2-N-[2-(4-acetylphenoxy)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)ureid-
o]phenyl]acetamido]ethylpiperazinyl-1-acetic acid
[1532] ##STR1953##
[1533] To a solution of
2-(N-benzyl-N-tert-butoxycarbonylamino)ethanol (5.90 g, 23.5 mmol),
4-hydroxyacetophenone (3.18 g, 23.5 mmol) and PPh.sub.3 (6.74 g,
25.8 mmol) in THF (100 mL) was added DIAD (5.20 mL, 25.8 mmol) at
room temperature The reaction mixture was heated under reflux for 4
hr and concentrated. Chromatography of the residue with
hexane-EtOAc (5:1, v/v) as eluent gave
4-[2-(N-benzyl-N-tert-butoxycarbonylamino)ethoxy]acetophenone (3.64
g, 42%) as a yellow oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.43-1.65
(m, 9H), 2.55 (s, 3H), 3.41-3.69 (m, 2H), 4.02-4.24 (m, 2H),
4.49-4.66 (m, 2H), 6.81-6.93 (m, 2H), 7.19-7.37 (m, 5H), 7.91 (d,
J=8.8 Hz, 2H); FAB-MS m/z 370 (M.sup.++1).
[1534] To a solution of
4-[2-(N-benzyl-N-tert-butoxycarbonylamino)ethoxy]acetophenone (3.05
g, 8.26 mmol) in CH.sub.2Cl.sub.2 (30 mL) was added TFA (20 mL) at
0.degree. C. After being stirred at room temperature for 2 hr, the
reaction mixture was concentrated. The residue was taken up with
sat. NaHCO.sub.3 solution and extracted with CHCl.sub.3. The
extracts were washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated to give 4-(2-N-benzylaminoethoxy)acetophenone (2.21 g,
99%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta. 2.04 (broad
s, 1H), 2.55 (s, 3H), 3.05 (t, J=5.4 Hz, 2H), 3.88 (s, 2H), 4.15
(t, J=5.4 Hz, 2H), 6.92 (d, J=8.9 Hz, 2H), 7.24-7.36 (m, 5H), 7.91
(d, J=8.9 Hz, 2H); FAB-MS m/z 270 (M.sup.++1).
[1535] To a solution of ethyl
4-(2-hydroxyethyl)piperazinyl-1-acetate (11.3 g, 52.1 mmol) and
CBr.sub.4 (20.7 g, 62.5 mmol) in CH.sub.2Cl.sub.2 (200 mL) was
added PPh.sub.3 (19.2 g, 73.0 mmol) portionwise at 0.degree. C. and
the reaction mixture was stirred for 30 min. Hexane was added to
the mixture, the precipitates were filtered off, and the filtrate
was concentrated to afford ethyl
4-(2-bromoethyl)piperazinyl-1-acetate (13.7 g, 94%) as a yellow
oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.27 (t, J=7.1 Hz, 3H),
2.61-2.78 (m, 8H), 2.81 (t, J=7.6 Hz, 2H), 3.20 (s, 2H), 3.42 (t,
J=7.6 Hz, 3H), 4.18 (q, J=7.1 Hz, 2H).
[1536] A mixture of 4-(2-N-benzylaminoethoxy)acetophenone (681 mg,
2.52 mmol), ethyl 4-(2-bromoethyl)piperazinyl-1-acetate (705 mg,
2.52 mmol), and K.sub.2CO.sub.3 (349 g, 2.52 mmol) in CH.sub.3CN
(10 mL) was heated under reflux for 22 hr. The resulting mixture
was filtered and the filtrate was concentrated to dryness.
Chromatography of the residue with toluene-acetone (1:1, v/v) as
eluent gave ethyl
4-[2-[N-benzyl-N-[2-(4-acetylphenoxy)ethyl]amino]ethylpiperazinyl-1-aceta-
te (920 mg, 78%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.27 (t, J=7.1 Hz, 3H), 2.45-2.53 (m, 10H), 2.55 (s, 3H), 2.76 (t,
J=6.8 Hz, 2H), 2.94 (t, J=6.1 Hz, 2H), 3.18 (s, 2H), 3.73 (s, 2H),
4.06 (t, J=6.1 Hz, 2H), 4.17 (q, J=7.1 Hz, 2H), 6.85 (d, J=7.1 Hz,
2H), 7.22-7.35 (m, 5H), 7.90 (d, J=7.1 Hz, 2H); FAB-MS m/z 468
(M.sup.++1).
[1537] A solution of ethyl
4-[2-[N-benzyl-N-[2-(4-acetylphenoxy)ethyl]amino]ethylpiperazinyl-1-aceta-
te (920 mg, 1.97 mmol) in EtOH (30 mL) was hydrogenated over 5%
Pd--C (53.1% wet, 510 mg) under hydrogen atmosphere for 4 hr. The
catalyst was filtered off and the filtrate was concentrated. The
residue was taken up with sat. NaHCO.sub.3 solution and extracted
with CHCl.sub.3. The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to afford ethyl
4-[2-N-[2-(4-acetyl phenoxy)ethyl]amino]ethylpiperazinyl-1-acetate
(690 mg, 93%) as a reddish oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.27 (t, J=7.1 Hz, 3H), 2.55 (s, 3H), 2.46-2.54 (m, 10H), 2.78 (t,
J=6.2 Hz, 2H), 3.04 (t, J=5.2 Hz, 2H), 3.20 (s, 2H), 4.13 (t, J=5.2
Hz, 2H), 4.18 (q, J=7.1 Hz, 2H), 6.92 (d, J=8.8 Hz, 2H), 7.92 (d,
J=8.8 Hz, 2H); FAB-MS m/z 378 (M.sup.++1).
[1538] To a solution of
3-methoxy-4-[N'-(2-methylphenyl)ureido]phenylacetic acid (558 mg,
1.77 mmol), ethyl
4-[2-N-[2-(4-acetylphenoxy)ethyl]amino]ethylpiperazinyl-1-acetate
(670 mg, 1.77 mmol), Et.sub.3N (0.38 mL, 2.66 mmol), and HOBt (50.0
mg, 0.35 mmol) in THF (10 mL) was added EDC.HCl (883 mg, 2.32 mmol)
at room temperature After stirring for 15 hr, the reaction mixture
was diluted with water and extracted with CHCl.sub.3-MeOH (10:1,
v/v). The extracts were washed with brine, dried over
Na.sub.2SO.sub.4, and concentrated to dryness. Chromatography of
the residue with CHCl.sub.3-MeOH (4:1, v/v) as eluent to give ethyl
4-[2-N-[2-(4-acetylphenoxy)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)urei-
do]phenyl]acetamido]ethylpiperazinyl-1-acetate (724 mg, 61%) as a
yellow amorphous solid. .sup.1H-NMR (CDCl.sub.3) .delta. 1.25, 1.27
(each t, J=7.1 Hz, total 3H), 2.29 (s, 3H), 2.45-2.51 (m, 8H), 2.54
(s, 3H), 2.55-2.63 (m, 2H), 3.17, 3.19 (each s, total 2H),
3.51-3.55 (m, 2H), 3.60 (s, 2H), 3.69 (s, 3H), 3.71-3.78 (m, 2H),
4.04-4.23 (m, 4H), 6.47 (m, 1H, J=8.1 Hz), 6.72-6.76 (m, 2H), 6.85
(d, J=8.8 Hz, 2H), 7.12-7.19 (m, 2H), 7.20-7.24 (m, 2H), 7.51 (d,
8.1 Hz, 1H), 7.88 (d, J=8.8 Hz, 2H), 8.04 (d, J=8.1 Hz, 1H); FAB-MS
m/z 674 (M.sup.++1).
[1539] To a solution of ethyl
4-[2-N-[2-(4-acetylphenoxy)ethyl]-N-[3-methoxy-4-[N'-(2-methylphenyl)urei-
do]phenyl]acetamido]ethylpiperazinyl-1-acetate (724 mg, 1.07 mmol)
in THF (10 mL) was added dropwise 0.25N NaOH (6.50 mL, 1.61 mmol)
at room temperature, and the reaction mixture was stirred for 1 hr.
The resulting mixture was concentrated, diluted with water, and
neutralized with 1N HCl at 0.degree. C. The mixture was extracted
with CHCl.sub.3-MeOH (4:1, v/v). The extracts were washed with
brine, dried over Na.sub.2SO.sub.4, and concentrated to dryness.
Chromatography of the residue with CHCl.sub.3-MeOH (3:1, v/v) as
eluent gave 331 (450 mg, 65%) as a pale yellow amorphous solid. IR
(KBr) 3345, 2938, 2821, 1673, 1631, 1598, 1533, 1455, 1417
cm.sup.-1; .sup.1H-NMR (DMSO-d.sub.6) .delta. 2.25 (s, 3H),
2.35-2.47 (m, 8H), 2.51 (s, 3H), 2.53-2.58 (m, 2H), 2.96 (s, 2H),
3.46-3.50 (m, 2H), 3.69 (s, 2H), 3.73-3.77 (m, 2H), 3.80, 3.83
(each s, total 3H), 4.19-4.22 (m, 2H), 6.73 (m, 1H), 6.85 (s, 1H),
6.92 (t, J=7.3 Hz, 1H), 7.30 (d, J=7.3 Hz, 2H), 7.10-7.16 (m, 2H),
7.79 (d, J=8.3 Hz, 1H), 7.90-7.95 (m, 2H), 8.00 (t, J=8.3 Hz, 1H),
8.55 (m, 1H), 8.61 (m, 1H); FAB-MS m/z 646 (M.sup.++1); Anal. Calcd
for C.sub.35H.sub.43N.sub.5O.sub.7.2HCl.H.sub.2O: C, 57.06; H,
6.43; N, 9.51. Found: C, 59.17; H, 6.32; N, 9.61.
Example 282
4-[2-N-[4-[N'-(2-bromophenyl)ureido]3-methoxyphenyl]-N-benzylacetamido]eth-
yl-1-piperidinylacetic acid
[1540] ##STR1954##
methyl
4-[2-N-[4-[N'-(2-bromophenyl)ureido]-3-methoxyphenyl]-N-benzylaceta-
mido]ethyl-1-piperidinylacetate
[1541] ##STR1955##
[1542] A mixture of piperidine ethanol (10.0 g, 77.4 mmol), benzyl
2-bromoacetate (17.8 g, 77.6 mmol) and K.sub.2CO.sub.3 (21.4 g, 155
mmol) in CH.sub.3CN (200 ml) was heated under reflux with stirring
for 2 h. The insoluble solid was removed by filtration, and the
filtrate was concentrated in vacuo. The residue was poured into ice
cooled 1N-HCl and extracted with CHCl.sub.3. The organic layer was
washed with water, drying over anhydrous Na.sub.2SO.sub.4, and
concentrated in vacuo. The residue was chromatographed on silica
gel with EtOAc as eluent to give benzyl
4-(2-hydroxyethyl)-1-piperidinylacetate (16.3 g, 76%) as a
colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta. 1.31-1.40 (m, 3H),
1.52 (dd, J=12.8, 6.3 Hz, 2H), 1.68 (brd, J=10.0 Hz, 3H), 2.16 (t,
J=11.6 Hz, 2H), 2.92 (brd, J=11.6 Hz, 2H), 3.24 (s, 2H), 3.69 (t,
J=6.8 Hz, 2H), 5.16 (s, 2H), 7.35 (m, 5H).
[1543] To a stirred solution of benzyl
4-(2-hydroxyethyl)-1-piperidinylacetate (14.9 g, 53.8 mmol) in
CH.sub.2Cl.sub.2 (150 ml) was added Et.sub.3N (37.5 ml, 269 mmol)
and DMSO (41.6 ml, 538 mmol). The reaction mixture was cooled to
0.degree. C. and SO.sub.3.Py (25.7 g, 161 mmol) was added portion
wise, and the resulting mixture was stirred at rt overnight. The
mixture was concentrated in vacuo, and the residue was diluted with
water, followed by extracted with Et.sub.2O. The organic layer was
washed with water, dried over anhydrous Na.sub.2CO.sub.3 and
concentrated in vacuo. The residue was chromatographed on silica
gel with EtOAc-hexane (2:1) as eluent to give (4-N-carbobenzyloxy
methylpiperidinyl)acetaldehyde (4.63 g, 31%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) .delta. 1.36-1.46 (m, 2H), 1.69 (br s,
2H), 1.72 (br s, 1H), 1.90 (m, 1H), 2.21 (dt, J=11.6, 2.4 Hz, 2H),
2.37 (dd, J=6.8, 1.6 Hz, 2H), 2.92 (d, J=11.6 Hz, 2H), 3.25 (s,
2H), 5.16 (s, 2H), 7.35 (m, 5H), 9.77 (t, J=2.0 Hz, 1H).
[1544] To a stirred solution of N-benzylamine (1.06 ml, 9.75 mmol)
in MeOH (20 ml) was added AcOH (560 ml, 9.75 mmol) and
(4-N-carbobenzyloxymethylpiperidinyl)acetaldehyde (1.79 g, 6.50
mmol) in MeOH (5 ml) and cooled to 0.degree. C. NaBH.sub.3CN (645
mg, 9.75 mmol) was added in one portion, and the resulting mixture
was stirred overnight at rt. The mixture was concentrated in vacuo,
and the residue was poured into aq.NaHCO.sub.3, then extracted with
CHCl.sub.3. The organic layer was washed with water, dried over
anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The residue
was chromatographed on silica gel with CHCl.sub.3-MeOH-EtOAc
(10:1:1) as eluent to give
N-benzyl-2-[4-(N-carbobenzyloxymethylpiperidinyl)]ethylamine (872
mg, 37%) as a colorless oil. .sup.1H-NMR (CDCl.sub.3) .delta.
1.36-1.46 (m, 2H), 1.69 (brs, 2H), 1.72 (brs, 1H), 1.90 (m, 1H),
2.21 (dt, J=11.6, 2.4 Hz, 2H), 2.37 (dd, J=6.8, 1.6 Hz, 2H), 2.92
(d, J=11.6 Hz, 2H), 3.25 (s, 2H), 5.16 (s, 2H), 7.35 (m, 5H), 9.77
(t, J=2.0 Hz, H).
[1545] To a stirred solution of
N-benzyl-2-[4-(N-carbobenzyloxymethylpiperidinyl)]ethylamine (356
mg, 0.972 mmol), 4-[N'-(2-bromophenyl)ureido]-3-methoxyphenylacetic
acid (369 mg, 0.972 mmol) and N,N-dimethylaminopyridine (119 mg,
0.972 mmol) in DMF (15 ml) was added EDC.HCl (372 mg, 1.94 mmol) at
rt, and the resulting mixture was stirred overnight. The reaction
mixture was poured into water and extracted with EtOAc. The organic
layer was washed with brine and dried over anhydrous
Na.sub.2SO.sub.4, then concentrated in vacuo. The residue was
chromatographed on silica gel with CHCl.sub.3-MeOH (10:1) as eluent
to give benzyl
4-[2-N-[4-[N'-(2-bromophenyl)ureido]3-methoxyphenyl]-N-benzylacetamido]et-
hyl-1-piperidinylacetate (611 mg, 86%) as a colorless oil.
.sup.1H-NMR (CDCl.sub.3) mixture of rotamars .delta. 1.15-2.09
(series of m, 7H), 2.06-2.14 (m, 2H), 2.84-2.96 (m, 2H), 3.17-3.23
(s, total 2H), 3.21 and 3.23 (s, total 2H), 3.38-3.42 (m, 1H), 3.65
and 3.73 (s, total 2H), 3.83 and 3.84 (s, total 3H), 4.49 (s, 1H),
4.60 (s, 1H), 5.15 (d, J=2.8 Hz, 2H), 6.74-7.83 (series of m, 16H),
7.51-7.53 (m, 1H), 7.94-8.02 (m, 1H), 8.18 (d, J=8.4 Hz, 2H); MS
(FAB) m/z 727 (M.sup.+), 729 (M.sup.++2).
[1546] To a stirred solution of benzyl
4-[2-N-[4-[N'-(2-bromophenyl)ureido]3-methoxyphenyl]-N-benzylacetamido]et-
hyl-1-piperidinylacetate (347 mg, 0.477 mmol) in MeOH--H.sub.2O
(5:1, 6 ml) was added LiOH (13.6 mg, 0.57 mmol) at rt, and the
resulting mixture was stirred for overnight. The reaction mixture
was poured into water and the solution was neutralized with aq.
1N-HCl, then extracted with CHCl.sub.3. The organic layer was
washed with brine and dried over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (10:1) as eluent to give 332 (97.3 mg,
32%) as a colorless amorphous solid, and 333 (168 mg, 54%) as a
colorless amorphous solid, respectively. 332 .sup.1H-NMR
(CD.sub.3OD), mixture of rotamars .delta. 1.29-1.90 (series of m,
7H), 2.75-2.86 (m, 2H), 3.28-3.51 (series of m, 6H), 3.74 and 3.78
(s, total 2H), 3.87 and 3.89 (s, total 3H), 4.66 (s, 1H), 4.85 (s,
1H), 6.79-7.00 (series of m, 3H), 7.16 (d, J=7.2 Hz, 1H), 7.23-7.37
(m, 5H), 7.57 (dd, J=8.4, 1.2 Hz, 1H), 7.88-8.00 (series of m, 2H);
MS (FAB) m/z 637 (M.sup.+), 639 (M.sup.++2). 333 .sup.1H-NMR
(CDCl.sub.3), mixture of rotamars a 1.15-2.11 (series of m, 7H),
2.89-2.97 (m, 2H), 3.17-3.49 (series of m, 6H), 3.17 and 3.18 (s,
total 3H), 3.70 and 3.71 (s, total 3H), 3.83 and 3.84 (s, total
2H), 4.49, 4.60 and 4.71 (s, total 2H), 6.75-8.16 (series of m,
14H); MS (FAB) m/z 651 (M.sup.+), 653 (M.sup.++2).
Example 283
4-[2-N-[4-[N'-(2-chlorophenyl)-3-methoxyureido]-phenyl]-N-benzylacetamido]-
ethyl-1-piperidinyl acetic acid
[1547] ##STR1956##
[1548] To a stirred solution of
N-benzyl-2-[4-(N-carbobenzyloxymethylpiperidinyl)]ethylamine (372
mg, 1.11 mmol),
4-[N'-(2-chlorophenyl)ureido]-3-methoxyphenyl]acetic acid (758 mg,
1.11 mmol) and N,N-dimethylaminopyridine (136 mg, 1.11 mmol) in DMF
(15 ml) was added EDC.HCl (426 mg, 2.22 mmol) at rt, and the
resulting mixture was stirred for 12 h. The reaction mixture was
poured into water and extracted with EtOAc. The organic layer was
washed with brine and dried over anhydrous Na.sub.2SO.sub.4, then
concentrated in vacuo. The residue was chromatographed on silica
gel with CHCl.sub.3-MeOH (10:1) as eluent to give benzyl
4-[2-N-[4-[N'-(2-chlorophenyl)-3-methoxyureido]phenyl]-N-benzylacetamido]-
ethyl-1-piperidinylacetate (668 mg, 88%) as a pale yellowish oil.
.sup.1H-NMR (CDCl.sub.3) mixture of rotamars .delta. 1.15-1.83
(series of in, 7H), 2.04-2.13 (m, 2H), 2.83-2.95 (m, 2H), 2.88 and
2.99 (s, total 2H), 3.20 and 3.23 (s, total 2H), 3.20-3.23
(overlap, 1H), 3.38-3.42 (m, 1H), 3.65-3.74 (m, 3H), 4.50 (s, 1H),
4.61 (s, 1H), 5.14 (d, J=4.4 Hz, 2H), 6.72-6.82 (series of m, 2H),
6.94-6.99 (m, 1H), 7.13-7.50 (series of m, 14H), 7.94-8.02 (m, 1H),
8.18 (d, J=8.0 Hz, 2H); MS (FAB) m/z 683 (M.sup.++H).
[1549] To a stirred solution of benzyl
4-[2-N-[4-[N'-(2-chlorophenyl)-3-methoxyureido]phenyl]-N-benzylacetamido]-
ethyl-1-piperidinylacetate (633 mg, 0.93 mmol) in MeOH--H.sub.2O
(10:1, 10 ml) was added LiOH (24.4 mg, 1.02 mmol) at rt, and the
resulting mixture was stirred overnight. The reaction mixture was
poured into water and the solution was neutralized with aq. 1N-HCl,
then extracted with CHCl.sub.3. The organic layer was washed with
brine and dried over anhydrous Na.sub.2SO.sub.4, then concentrated
in vacuo. The residue was chromatographed on silica gel with
CHCl.sub.3-MeOH (20:1) as eluent to give methyl
4-[2-N-[4-[1N'-(2-chlorophenyl)-3-methoxyureido]phenyl]-N-benzylacetamido-
]ethyl-1-piperidinylacetate (504 mg, 89%) as a colorless amorphous
solid. .sup.1H-NMR (CDCl.sub.3), mixture of rotamars .delta.
1.26-1.64 (series of m, 7H), 2.02-2.10 (m, 2H), 2.86 (t, J=10.4 Hz,
2H), 3.17 (d, J=8.0 Hz, 2H), 3.16-3.22 (m, overlap, 1H), 3.40 (t,
J=7.6 Hz, 1H), 3.48 (s, 1H), 3.65 and 3.73 (s, total 2H), 3.70 and
3.71 (s, total 3H), 3.79 and 3.80 (s, total 3H), 4.50, 4.60 and
4.70 (s, total 2H), 6.73-6.85 (series of m, 2H), 6.98 (t, J=7.6 Hz,
1H), 7.13-7.37 (series of m, 9H), 7.96 (m, 1H), 8.18 (d, J=8.0 Hz,
2H); MS (FAB) m/z 607 (M.sup.++H).
[1550] To a stirred solution of methyl
4-[2-N-[4-[N'-(2-chlorophenyl)-3-methoxyureido]phenyl]-N-benzylacetamido]-
ethyl-1-piperidinylacetate (177 mg, 0.292 mmol) in MeOH--H.sub.2O
(5:1, 6 ml) was added LiOH (21.6 mg, 0.90 mmol), and the resulting
mixture was stirred for 4 h at rt. The mixture was poured into
water, and the solution was neutralized with aq. 1N-HCl, then
extracted with CHCl.sub.3. The organic layer was washed with brine
and dried over anhydrous Na.sub.2SO.sub.4, then concentrated to
yield 334 (155 mg, 92%) as a colorless amorphous solid. .sup.1H-NMR
(CD.sub.3OD) mixture of rotamars .delta. 1.28-1.90 (series of m,
7H), 2.84 (brs, 2H), 3.30-3.52 (series of m, 6H), 3.74 and 3.82 (s,
total 2H), 3.87 and 3.88 (s, total 3H), 4.63 (s, 1H), 4.66 (s, 1H),
6.79-7.05 (series of m, 3H), 7.16 (d, J=7.2 Hz, 1H), 7.24-7.40 (m,
5H), 7.88 (s, 1H), 7.98-8.04 (series of m, 2H); MS (ESI) m/z 593
M.sup.+), 615 (M.sup.++Na.sup.+).
[1551] It should be understood that while this invention has been
described herein in terms of specific embodiments set forth in
detail, such embodiments are presented by way of illustration of
general principles, and the invention is not necessarily limited
thereto. Modifications and variations in any given material or
process step will be readily apparent to those skilled in the art
without departing from the true spirit and scope of the following
claims, and all such modifications are included within the scope of
the present invention.
* * * * *