AU764553B2 - Inhibitors of alpha 4 beta 1 mediated cell adhesion - Google Patents

Inhibitors of alpha 4 beta 1 mediated cell adhesion Download PDF

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Publication number
AU764553B2
AU764553B2 AU47116/99A AU4711699A AU764553B2 AU 764553 B2 AU764553 B2 AU 764553B2 AU 47116/99 A AU47116/99 A AU 47116/99A AU 4711699 A AU4711699 A AU 4711699A AU 764553 B2 AU764553 B2 AU 764553B2
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alkyl
hydrogen
attached
carbon atom
formula
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AU4711699A (en
Inventor
James R. Blinn
Robert A Chrusciel
Jed F. Fisher
Thomas J. Lobl
Steven P. Tanis
Bradley R. Teegarden
Edward William Thomas
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Tanabe Seiyaku Co Ltd
Pharmacia and Upjohn Co
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Tanabe Seiyaku Co Ltd
Pharmacia and Upjohn Co
Upjohn Co
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Description

WO 99/67230 PCT/US99/14233 -1- INHIBITORS OF 4, MEDIATED CELL ADHESION Background of the Invention Field of the Invention The present invention relates to small molecules that are potent inhibitors of ao43 mediated adhesion to either VCAM or CS-1 and which are useful for the treatment of inflammatory diseases.
Description of Related Art The extracellular matrix (ECM) is the major component of connective tissue which provides structural integrity, and promotes cell migration and differentiation. As part of these functions, extracellular matrix molecules such as fibronectin, collagen, laminin, von Willebrand factor, thrombospondin, fibrinogen, and tenascin have been shown to support adhesion of cells in vitro. This adhesive interaction is critical for a number of biological processes including hemostasis, thrombosis, wound healing, tumor metastasis, immunity and inflammation.
Fibronectin (FN) is the prototype ECM molecule. The major cell attachment site in the fibronectin molecule has been reproduced synthetically with the amino acid sequence arginine-glycine-aspartic acid, or RGD using single letter nomenclature. Peptides containing the RGD sequence which either inhibit or promote cell adhesion have been described (US Patent Nos. 4,589,881; 4,661,111; 4,517,686; 4,683,291; 4,578,079; 4,614,517; and 4,792,525). Changes in the peptide as small as the exchange of alanine for glycine or glutamic acid for aspartic acid, which constitute the addition of a single methyl or methylene group to the tripeptide, eliminates these activities (Pierschbacher et al., Proc. Natl. Acad. Sci.
USA 81:5985 (1984)). Recently, a second FN cell binding domain has been identified within the alternatively spliced region of the A chain of the molecule, known as the connecting segment 1 The most active cell-binding site within this alternatively spliced region is composed of 25 amino acids where the carboxy terminus contains the sequence WO 99/67230 PCT/US99/14233 -2- EILDVPST. The amino acid sequence EILDVPST forms a recognition motif on FN for cell surface receptors. (Wayner et al., J. Cell Biol. 109:1321 (1989); Guan et al., Cell 60:53 (1990)).
The receptors which recognize these sites on FN belong to a gene superfamily called integrins which consist of heterodimeric complexes of non-covalently associated alpha and beta subunits. A common p subunit combines with unique I subunits to form an adhesion receptor of defined specificity. To date, 8 P subunits have been identified which can dimerize with 16 distinct I subunits forming 22 distinct integrins. The 131 subfamily, also known as the VLA family (Very Late Activation Antigens), binds to ECM molecules such as FN, collagen and laminin. For reviews, see, Hynes, Cell 48:549 (1987); Hemler, Annu. Rev.
Immunol. 8:365 (1990). Leukocyte interaction with FN at the two spatially separate binding domains is mediated by two distinct integrins. The RGD site is recognized by the integrin as,3, while, EILDV is recognized by a4p, (Pytela et al., Cell 40:191 (1985); Wayner et al., J.
Cell Biol. 109:1321 (1989); Guan et al, Cell 60:53 (1990)).
Vascular endothelial cells form the interface between blood and tissues and control the passage of leukocytes as well as plasma fluid into tissues. A variety of signals generated at the site of inflammation can activate both endothelial cells as well as circulating leukocytes so that they become more adhesive to one another. Following this initial adhesion the leukocytes migrate into the tissues to perform host defense functions. Several adhesion molecules have been identified which are involved in leukocyte-endothelial interactions.
In the 3, subfamily, in addition to binding to fibronectin, ca4, interacts with a cytokine inducible protein on endothelial cells termed vascular cell adhesion molecule (VCAM). Further involved in the leukocyte-endothelial adhesion process is the P1 integrin subfamily. 32 integrins include CDlla/CD18, CDllb/CD18, and CDllc/CD18. In addition, the P, subunit associates with a 4 to form a unique c 4 P, heterodimer which binds to FN, to VCAM, and to Mucosal Addressin Cell Adhesion Molecule-1 (MAdCAM) (Ruegg et al, J. Cell.Biol. 117:179 (1992); Andrew et al., J Immunol. 153:3847 (1994); Briskin et al., Nature 363:461 (1993); Shyjan et al, J. Immunol. 156:2851 (1996)). a 4 integrins are widely expressed on different cell types including hematopoietic progenitors, lymphocytes, natural
I
WO 99/67230 PCT/US99/14233 -3killer cells, monocytes, eosinophils, basophils, and mast cells (Helmer, M. Annu. Rev.
Immunol. 8:365 (1990)). Other molecules on endothelial cells which bind to the leukocytes include ICAM-1, ICAM-2, E-selectin and P-selectin (Carlos and Harlan, Immunol. Rev.
114:1 (1990); Osbom, Cell 62:3 (1990); Springer Nature 346:425 (1990); Geng et al., Nature 347:757 (1990); Stoolman, Cell 56:907 (1989)).
A number of in vitro and in vivo studies indicate that a 4 I plays a critical role in the pathogenesis of a variety of diseases. Monoclonal antibodies directed against a 4 have been tested in a variety of disease models. Anti-a 4 antibodies block adhesion of lymphocytes to synovial endothelial cells; this adhesion plays a potential role in rheumatoid arthritis (van Dinther-Janssen et al, J. Immunol. 147:4207 (1991)). 4a has also been implicated with respect to rheumatoid arthritis in separate studies (Laffon et al, J. Clin. Invest. 88:546 (1991); Morales-Ducret et al, J Immunol. 149:1424 (1992)). A significant number of studies have evaluated the role of a 4 in allergy and asthma. For example, monoclonal antibodies to a 4 block adhesion of basophils and eosinophils to cytokine activated endothelial cells (Walsh et al, J. Immunol. 146:3419 (1991); Bochner et al, J. Exp. Med. 173:1553 (1991)).
Monoclonal antibodies to a 4 were also effective in several lung antigen challenge models (Abraham et al, J Clin. Invest. 93:776 (1994); Weg et al, J Exp. Med 177:561 (1993)).
The cotton-top tamarin, which experiences spontaneous chronic colitis, showed a significant attenuation of their colitis when anti-c, antibody was administered (Podolsky et al, J. Clin.
Invest. 92:372 (1993); Bell et al, J. Immunol. 151:4790 (1993)). In a rat and mouse model, autoimmune encephalomyelitis was blocked by anti-a, antibody (Yednock et al, Nature 356:63 (1992); Baron et al, J. Exp. Med. 177:57 (1993)). Anti-a, monoclonal antibodies also inhibit insulitis and delay the onset of diabetes in the non-obese diabetic mouse (Baron et al, J. Clin. Invest. 93:1700 (1994); Yang et al, Proc. Natl. Acad. Sci. USA 90:10494 (1993); Burkly et al, Diabetes 43:529 (1994)). ac 4 is also implicated in atherosclerosis due to its endothelial expression during atherogenesis (Cybulsky et al, Science 251:788 (1991)).
The migration of leukocytes to an inflammatory site can also be blocked by anti-a 4 antibodies. In addition to the blocking of migration, inhibitors of leukocyte endothelial adhesion may block the costimulatory signals mediated by integrins and thus inhibit overproduction of inflammatory cytokines. In a separate set of experiments not using anti-a 4 antibodies, the peptides GRDGSP or EILDV were tested against contact hypersensitivity response. The contact hypersensitivity response was found to be blocked by GRDGSP or EILDV suggesting that both afp, and 5 are involved in this inflammatory response.
Other ailments which may involve a4P,-mediated conditions include the inflammatory disorders rheumatoid arthritis, allergic disorders, asthma, spontaneous chronic colitis, insulitis, contact hypersensitivity response, atherosclerosis and autoimmune encephalomyelitis. These studies illustrate that small molecules that are potent inhibitors of a4p, mediated adhesion to either VCAM-I or CS-1 may be used as a form of treatment in numerous inflammatory diseases. However, these inflammatory conditions could be expanded to include adult respiratory distress syndrome. AIDS, cardiovascular diseases, thrombosis or harmful platelet aggregation, reocclusion following thrombolysis, allograft rejection, reperfusion injury, psoriasis, eczema, contact dermatitis and other skin inflammatory diseases, osteoporosis, osteoarthritis, atherosclerosis, neoplastic diseases including metastasis of neoplastic or cancerous growth, wound healing enhancement, treatment of certain eye diseases such as detaching retina, Type I diabetes, multiple sclerosis, systemic lupus erythematosus (SLE), inflammatory and immunoinflammatory conditions including ophthalmic inflammatory conditions and inflammatory bowel diseases, ulcerative colitis, regional enteritis and other autoimmune diseases. Accordingly, a compound which could inhibit these conditions is desirable.
The discussion of the background to the invention herein is included to explain the context of the invention. This is not to be taken as admission that any of the material referred to was published, known or part of the common general knowledge in Australia as at the priority date of any of the claims.
Summary of the Invention In one aspect the present invention provides a compound of the formula:
R
2
H
N )m Rs
R
3 -Y n
O
wherein R, may occur one to four times and each occurrence is independently hydrogen or
C,
4 alkyl; R, is hydrogen, pyridyl, C, 6 alkyl, (C, 4 alkyl)-CO,-R,,, or in addition, R, and R. may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: /(CH2)N C NRII (CH2) 10 together with the carbon atom to which they are attached: R, is hydrogen. phenyl, C, 4 alkyl, C 3 4 alkenyl, arylalkyl (C, 4 alkyl)-CO 2
(C
2 alkenyl)-CO,-R,,, alkyl)-CO-C, 4 alkyl, alkyl)-O-C,I alkyl, (C, 4 alkyl)- OH, alkyl)-CN, adamantyl or one of the following: 1-6 aflyo,-K -1 0 5
~O
H-N
H
1-6 alky)--N Z -N Z C0 2
-RI
1 C0 2
-RII
N
-C C 4-02-Rit -(C-6aN
O-
3 alcy)] 13 -&02-RI 0- N
OH
0 -o C-c
F
3 C9
N
N(C
1 aky) 2 H -H3 H C02-RI I H C0-RI 0 N02
HN-N
"A'S-C0 2
-R
in addition, R 2 and may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: R, is NH 2 NHOH, 10 arylalkyl), or is of the formula S-o-CHz--27)
N;
R, is a formula of the following: r
-R
8
-R
12 R. is N or CH; R, is hydrogen or halogen; R, is -NH-Y, or -CONH-; R9 may occur one to thiree times and is a halogen, C, alkoxy, C,.6 alkyl or trifluoromethyl; Rio is C, alkyl. or alkyl)-OH, or hydrogen; is hydrogen or alkyl; R1 2 is alkyl or the following formnula:
R
13
-CH
2 )r
R
is N or CH; W is(CI., alkyl); X is S; Y and Y, are independently or Z is 0, CH,, or NR, 1 s 1, 2 ,or 3; m is I or 2; n is 0 or 1; o isO0 or 1; p isO0 or 1; q is 0or 1; and r is0, 1, 2 or 3; with the provisos that when Y is R, cannot be hydrogen; and when R, is equal to alkyl), C, alkvl is not equai (o fert-butyl.
In another aspect the present invention provides a compound of the flormula: 2 0
N
R (X R Rr 'N n m
R
0 0 wherein R, may occur one to four times and each occurrence is independently hydrogen or C,.6 alkyl; R, is hydrogen, pyridyl, C,.6 alkyl, (C 1 6 alkyl)-C0 2 or CR; in addition, R, and R, may be attached to thie same carbon atom and form a carbocyclic.
ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: together with the carbon atom to which they are attached; R(3 is hydrogen, phenyl, C, alkyl, CM. alkenyl, arylalkyl (C, 4 6 alkyl)-C0 2
R
11 (C7.6 alkenyl)-C0 2
(C,
4 6 alkyl)-CO-C, 14 alkyl, alkyl)-O-C, 4 6 alkyl, alkyl)- OH, alkyl)-CN, adamantyl or one of the following: N-.N "n o OR,, -(G16a)-(ia N-N0
H
1-6 akyl)-N Z -N Z
N
C.
a
,OCI
3 aky N1- C0-RI, N02Ri -(C-6N
OH
0 -0 4F 3
N(C,.
3 akylh S-0 S J- H C0 2 -Rjj H C0R 1 0 NO,
N
5 V -C02R,, -4ein addition, R, and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together wi~th the carbon atom and the nitrogen atom to which they are attached:
ZN,
P,4 is -O-RI NH 2 NHOH, arylalkyl), or is of the formula R, is a formula of the following: p
RR,
Ris N or CH; R, is hydrogen or halogen; R, is or -CONH-; R.0 may occur one to three times and is a halogen, alkoxy, C,, 6 alkyl or trifluoromethyl; is C, alkyl, or alkyl)-OH, or hydrogen; R, is hydrogen or C,_6 alkyl; R, is alkyl or the following formnula:
(CH
2 R(9 111 3 is N or CH; W is alkyl); X Is S; Y and Y, are independently
-SO
2 or -C(=O)N(Ri 0 y-; Z is0, CH 2 or NR,.
/is 1, 2 ,or3; m Is I or 2; *nis 0or 1; is 0 or 1; pis 0or 1; q is 0 or 1; and r isO0, 1. 2 or 3-, with thc provisos that (1 when Y is R, cannot bc hydrogen: wheni R, is equal to alkyl), C, alkyl is not equal to tert-butyl; and the compoundc has an lC, 0 value oflcss than SuM in a Jurkat CS- I assay and/or an 1CSO value of less than S50 1 in f a Jurkat [C assay.
In another aspect the present invention provides a compound of the formula: R2 H 0
N
R
1 1 N )m
R
0 0 wherein R, may occur one to four times and each occurrence is independently hydrogen or alkyl;
R
2 is hydrogen, pyridyl, C,.6 alkyl, 6 alkyl)-C0 2 -RI I, or -C0 2 in addition, R, and R, may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula:
(CH
2 )1\
CH
together with the carbon atom to which they are attached; R, is hydrogen, phenyl, C, alkyl, C 3 6 alkenyl, arylalkyl alkyl)-COf-
R,
1 alkcnyl)-C0 2
-R
1 6 alkyl)-CO-C,, alkyl, alkyl)-O-C 1 6 alkyl, alkyl)- OH, alkcyl)-CN, adamantyl or one of the following: N-N >o RI akyI--K N N N
H
-(CI-6akcyr)-N Z f (V N C0 2 -RII C2RI 0-i C 6
NN
N19 IO(CI..
3 alkyl.
3 0-RI
OH
0 _0 N
F
3 N(C 3 akYi) 2 H C0 2
-RI
1 H C0 2 -R I 0 N1
HN-N
N >C0 2
-R
in addition, R 2 and R 3 may combine vith each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N.
R, is NH2, NHOH, arylatkyl), or is of the formula R, is a formula of the following: a.
S
a R,is N or CH; R, is hydrogen or halogen; R, is -NHCO-; P, may occur one to three times and is a halogen, C,.
6 alkoxy, C, 4 alkyl or tnifluoromethyl; RIO is alkyl, or 6 alkyl)-OH, or hydrogen; is hydrogen or C, alkcyl; R1 2 iS C1.6 alkyl or the following formula: -CH2)r 4 K<R9 RD~ is N or CH; W is (C, 4 alkyl); X is S; Y is or Z is 0, Cl- 2 or N-R,; I s 1, 2, or 3; m is I or 2; n is 0or 1; o is 0or 1; p is 0or 1; q is 0or 1; and r isO0, 1, 2 or 3; with the provisos that when Y is R 3 cannot b6 hydrogen; and when R, is equal to O-(C 4 alkyl), C, alkyl is not equal to tert-butyl.
In another aspect the present invention provides a compound of the formula: R2 H 0
N
RI (X R3- y'N M
R
0 0 wherein R, may occur one to four times and each occurrence is independently hydrogen or alkyl; is hydrogen, pyridyl, alkyl, alkyl)-C0 2 -RI I, or -C0 2 1 in addition, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: *C /NR 11
I
(CH
2 10 together with the carbon atom to wh-ich they are attached; R,3 is hydrogen, phenyl, alkyl, C3. alkenyl, 7 arylalkyl alkyl)-C0 2 R, 1 (C2-6 alkenyl)-CO,-R, alkyl)-CO-CI., alkyl, (CI.6 alkyl)-O-C -6 alkyl, alkyl)- OH, alkcyl)-CN, adamantyl or one of the following: N- N ORI I
H
-(C1-6 aky--N Z -N Z 'kN C 2 -R 1 C0 2
-R
11 02-Rii (CI-6ak
N
NG (-aC- Q C 2
-R
1
OH
0 -0
-I--N-CF
3
N(C..
3 alcl) H C0 2
-R
1 H C0 2
-RI,
0
NO
2
HN-N
C 2 -Rj in addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N\ z 9 99** *99* R, is NH 2 NHOH, arylalkyl), or is of the formula -o-CH 2 {14'
N*
R, is a formula of the following: .9 *9 9* 9 Ris N or CH; R, is hydrogen or halogen; It, is -NHCO-;
R
9 may occur one to three times and is a halogen, C,.
6 alkoxy, alkyl or trifluoromethyl; Rio is C, alkyl, or alkyl)-OH, or hydrogen; R, I is hydrogen or C,,6 alkyl; is alkyl or the following formula:
-CH
2 )r
R
is N or CH; W iS (CI.6 alkyl); X is S; Y is or -(ONRo- Z isO0, CH., or N-R,,1; I s 1, 2, or 3; m is Ior 2; n isO0 or 1; o is 0or 1; 4pis 0or 1; *q 0is 0or 1; and r is0,1, 2 or 3; with the provisos that when Y is R 3 cannot be hydrogen; when R, is equal to alkyl), C, alkyl is not equal to tert-butyl; and 0 *000"(3) the compondc has an value of less tha-n 5iaM in a Jurkat CS- I assay 4...and/or an IC,, value of less tha-n 50iaM in a Jurkat EC assay.
4.04.
a4 0 -4m- The present invention particularly provides: A compound of the formula: P" H 0 R, X w(R4 0 0 **so S.f sees0 :so.
foe sees goo& WO 99/67230 WO 9967230PCTIUS99/1 4233 In the above formula R, may occur one to four times and each occurrence is independently hydrogen or C,- 6 alkyl. Also in the above formula R 2 is hydrogen, pyridyl, C 1 6 alkyl, (C 1 6 alkyl)-C0 2 or -C0 2 In addition, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula:
(CH
2 )1/ together with the carbon atom to which they are attached.
In the above formula R 3 is hydrogen, phenyl, C,- 6 alkyl, C 3 6 alkenyl, C1.
17 arylalkyl I (C 1 6 alkyl)-C0 2
(C
2 6 alkenyl)-C0 2
(C
1 6 alkyl)-CO-C,- 6 alkyl, 6 alkyl)-O-C,.
6 alkyl, 6 alkyl)-OH, 6 alkyl)-CN, adamantyl or one of the following: 1 -6 aky )-KI \O N-N 0
H
-(CI -6 alkyi)-N \Z -N Z C0 2 -RII N C0 2 Rj 1 C02-Rjj 6 akyl+)- -fl-O(C,- 3 a~ky0]1. 3 -GC02-R,
I
N
OH
0 -0
CF
3 4
N
N
H C0 2
I
S
H C0 2
I
0
NO,)
HN-N
C0 2 -Rj I N(C 1- 3 alky) 2 In addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: WO 99/67230 PCT/US99/14233 -6- Z N
O
0 In the above formula R 4 is NH 2 NHOH, 0 arylalkyl), or is of the formula: -O-CH2 In the above formula R, is a formula of the following: R8- R I2 R7 In the above, R 6 is N or CH, R, is hydrogen or halogen, R 8 is -OCH 2 or -CONH-, R 9 may occur one to three times and is a halogen, alkoxy, alkyl or trifluoromethyl, Ri 0 is alkyl, or alkyl)-OH, or hydrogen, R, is hydrogen or C,6 alkyl, R, 2 is C,6 alkyl or the following formula:
R
1 3
(CH
2 )r R 9 and R, 3 is N or CH.
In the above formula W is 6 alkyl), X is S, O, or CH 2 Y and Y, are independently
-SO
2 or 0 and Z is O, CH,, or In the above, I is 1, 2, or 3, m is 1 or 2, n is 0 or 1, o is 0 or l,p is 0 or 1, q is 0 or 1, andr is 0, 1, 2 or 3.
The above formula has the provisos that: when Y is R 3 cannot be hydrogen; when R 4 is equal to O-(C 4 alkyl), C 4 alkyl is not equal to tert-butyl; in those pyrrolidine structures (1 is 1; m is 2; n is 0; o is 0; p is 1; X is
CH
2 W is equal to CH,; and WO 99/67230 PCT/US99/14233 -7the compound is not H O qOzH S NCl In another embodiment of the present invention, R 5 is a formula of the following: ci
RC
R7
CI
wherein R, is hydrogen or Cl.
In another embodiment of the present invention, R 2 is hydrogen or alkyl.
Additionally, R, and R 2 may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached or be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: C /(CH 2
I
C NRI1 \(CH2)/ together with the carbon atom to which they are attached. Also, in this embodiment, n is 0, m is 2 andp is 0.
In yet another embodiment of the present invention, R, is hydrogen or C 13 alkyl, and R 2 is hydrogen or alkyl. Additionally, R, and R, may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached or be attached to the same atom and form a ring of 5-8 atoms of the formula: /(CH2)\ (CH2)/ WO 99/67230 WO 9967230PCTIUS99/1 4233 -8together with the carbon atom to which they are attached. Also in this embodiment, R 3 is hydrogen, C,- 6 alkyl, C 717 arylalkyl, (C 6 alkyl)-OH, (C 6 alkyl)-CO,-R, (C 1 6 alkyl)-CN, adamantyl, phenyl, or one of the following: Nl)_ N
NN
H
-6 alkyl)-N Z N Z
N(CH
3 2 Additionally, in this embodiment, R 4 is NH 2 NHOH, or R 4 is of the formula:
CH
2 N R, I is hydrogen or CH 3 and X is S or 0.
In another embodiment, R, and R 2 are hydrogen, and R 3 is (C 2 6 alkenyl)-C0 2
-R,
(C
1 6 alkyl)-O-C.
3 alkyl, (C 6 alkyl)-C0 2 R, or one of the following:
N~
T -C0 2 0
N
0 N02HN-N N 0.
1 jj.-R11 Also, in this embodiment, R 4 is (6 is CH. is hydrogen, and R(7 is hydrogen.
Additionally, X is S, Y is and 1 is 1.
In another embodiment of the present invention, R, and R 2 are hydrogen, and R 3 is
C
1 6 alkyl, (C 1 6 alkyl)-C0 2 1
(C
2 6 alkenyl)-C0 2 (C -6 alkyl)-CO-C,- 6 alkyl,
(C
1 6 alkyl)-O-C,.
3 alkyl, (C 16 alkyl)-CN, or one of the following: WO 99/67230 WO 9967230PCT/US99/I 4233 -9- N. C0 2
-R
1 I C0 2 -RI I C0 2 -RI I 16 Ally N -0aN
N
N
N. 0 2 I
CF
3
OCH
3 N
OCH
3
OHN
SS NO0H-
OR]
1 -N N CO
N'C
2 -R n H: C0 2 -RjI H C0 2 02R Additionally, R,4 is OH, R, is CH, R, is hydrogen, R(7 is hydrogen, X is CH 2 and Y is CO- or In another embodiment of the present invention, W is (C 3 alkyl), X is CH 2 Y is R, is hydrogen, R 2 is hydrogen, (C 3 alkyl)-C0 2 or -C0 2 1
R
3 is hydrogen, C 7 10 arylalkyl, C,- 6 alkyl, or 6 alkyl)-C0 2 ,1R4 is OH, R6 is CH, R, is hydrogen, R, is hydrogen, 1 is I or 3, and n is 0.
In another embodiment of the present invention, W is C,- 3 alkyl, X is CH,,R, is hydrogen, R 2 is (C 4 alkyl)-C0 2 or C0 2
-R
1 1(3 is hydrogen, C 1.
3 alkyl, or C 2 7 alkenyl, R(4 is OH, R6 is CH, is hydrogen, R, is hydrogen, 1 is 1, m is 1, n is 1, o isO0, and p is 1.
In another embodiment, the compound of the present invention is represented by the following formula f-S NH 0
R
In another embodiment of the present invention, R, and R. are hydrogen, and R(3 is C 1.
6 alkyl, (C 1 6 alkyl)-C0 2 R, (C 2 6 alkenyl)-C0 2 R, (C 6 alkyl)-O-C 3 alkyl, or one of the following: WO 99/67230 PCT/US99/14233 -in C0 2 -R, K-
OH
HN-N
HCO2-R,, CO2R, Additionally, in this embodiment, R 4 is R 6 is CH, is hydrogen or C,.
6 alkyl, R, is hydrogen, X is S, Y is and 1 is 1.
In another embodiment of the present invention, R, is hydrogen or C.
3 alkyl and R 2 is hydrogen or C-4 alkyl. Additionally, R, and R, may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms, or be attached to the same atom and form a ring of 5-8 atoms of the formula:
/(CH
2 C NRI \(CH2)/ together with the carbon atom to which they are attached. In this embodiment, R 3 is hydrogen, C,.
6 alkyl, C., 7 arylalkyl, alkyl)-OH, 6 alkyl)-CO2-R,,, 6 alkyl)-CN, adamantyl, phenyl, or one of the following:
N
-6 N
N
1-6 alky
N
-N Z
<V
N(C l-3aOkyD 2 -6 alkyl)-N Z CF3 n
CO
2
RI,
NO
2
OH
OH
Additionally, in this embodiment, R 4 is NH,, NHOH, or R 4 is of the formula WO 99/67230 PCT/US99/14233 -11- -O-CH2- N Also, is hydrogen or C, 6 alkyl, and X is S or O.
In yet another embodiment, the compound of the present invention is represented by the following formula R2 H O R, W RN (I-a) N )m R 0 0 In the above formula R, may occur one to four times and each occurrence is independently hydrogen or C, 6 alkyl, and R 2 is hydrogen, pyridyl, C,6 alkyl, alkyl)-
CO
2 or -CO 2 Additionally, R, and R2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: C C NRI (CH2) together with the carbon atom to which they are attached. Additionally, in this embodiment, R 3 is hydrogen, phenyl, C, 6 alkyl, C 36 alkenyl, arylalkyl 6 alkyl)-CO2-R,,, (C2- alkenyl)-CO2-R,,, 6 alkyl)-CO-C,.
6 alkyl, (C, 6 alkyl)-O-C,.
6 alkyl, alkyl)-OH, (C, 6 alkyl)-CN, adamantyl or one of the following: WO 99/67230 WO 9967230PCT[US99/I 4233 ON.N 5y\OR 11 1 6 aky)- -1
H
N C0 2
-R
1 I C0 2 -RI I~
N
-(C
1 6 alky -N Z q -N Z
N
N NC0 2 -Rjj 1.6al
N
[,-GO(C
1 3 alkyDI)J 3
CN
OH
0 _0
CF
3
N
<S C 2R
I
H
Nl H C0 2
I
0 NO-, HN-N K-1 O-j N(C 1 3 alky) 2 In addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N 0 Additionally, R, is NH- 2 NHOH, o arylalkyl), or is of the formula -0-CN
N*
Also, in this embodiment, R, is a formula of the following: Additionally, in this embodiment, R 6 is N or CH, R 7 is hydrogen or halogen, R, is WO 99/67230 PCT/US99/14233 -13-
-OCH
2 or -CONH-, R 9 may occur one to three times and is a halogen, C,6 alkoxy, C,6 alkyl or trifluoromethyl, Rio is C,- 6 alkyl, or 6 alkyl)-OH, or hydrogen, R,, is hydrogen or C,, 6 alkyl, R 12 is C- 6 alkyl or the following formula: R13
-(CH
2 )r
R
9 R3 is N or CH, W is 6 alkyl), X is S, O, or CH 2 Y and Y, are independently -SO2-, or Z is O, CH 2 or 1 is 1, 2, or 3, m is 1 or2, nis0or, ois0or ,pis 0 or 1, q is 0 or 1 and r is 0, 1,2 or 3. This particular embodiment of the present invention has the provisos that when Y is R 3 cannot be hydrogen; when R 4 is equal to O-(C 4 alkyl), C 4 alkyl is not equal to tert-butyl; in those pyrrolidine structures (1 is 1; m is 2; n is 0; o is 0; p is 1; X is
CH
2 W is equal to CH,; the compound has an ICs value of less than 5uiM in a Jurkat CS-1 assay and/or an IC, 5 value of less than 50M in a Jurkat EC assay; and the compound is not Cl H 0 CO 2 H 0 H
O
In yet another embodiment of the compound of formula R, may occur one to four times and each occurrence is independently hydrogen or C,.
6 alkyl. Also, R 2 is hydrogen, pyridyl, C,.
6 alkyl, alkyl)-CO 2 or Additionally, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: WO 99/67230 WO 9967230PCTIUS99/1 4233 -14together with the carbon atom to which they are attached. In this embodiment, R 3 is hydrogen, phenyl, C, 6 alkyl, C 36 alkenyl, C 7 arylalkyl (C 6 alkyl)-C0 2
(C
2 6 alkenyl)-C0 2 1, (C 1 6 alkyl)-CO-C,-6 alkyl, (C 1-6 alkyl)-O-C- 6 alkyl, (C- 6 alkyl)-OH, alkyl)-CN, adamantyl or one of the following:
H
16 aky)- N z q -N ZtN N.C0 2 -Rl I
(O(CI-
3 alkyo], -3 N-)C0 2 -Ri
I
7C j C0 2 -Rjj 6 akyI)
N
N.
GH)C0 2 -Rjj
I
1
OH
0 N -CF 3 H- S C 0 2 -R
S
H CO-,-Rj I 0
NO-)
HN-N
C0 2 -Rj I N(C 3 alkyl), In addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: zN In this embodiment, R 4 is 1, NH 2 NHOH, -O-(C 7 arylalkyl), or R 4 is of the formula: WO 99/67230 PCT/US99/14233 o -CH2-thit, Also, in this embodiment, R, is a formula of the following: Additionally, in this embodiment, R 6 is N or CH, R, is hydrogen or halogen, R, is -NHCO-, R, may occur one to three times and is a halogen, Ci.6 alkoxy, C, 6 alkyl or trifluoromethyl, R,o is C,.
6 alkyl, or 6 alkyl)-OH. or hydrogen, is hydrogen or C 6 alkyl, R2, is C,.
6 alkyl or the following formula: R,3 is N or CH, W is (C 6 alkyl), X is S, O, or CH2, Y is -CO-, Z is O, CH 2 or 1 is 1, 2, or 3, m is 1 or 2, n is 0 or 1, o is 0 or 1, p is 0 or 1, q is 0 or 1, and r is 0, 1, 2 or 3. In this embodiment, the following provisos apply: when Y is R 3 cannot be hydrogen; when R 4 is equal to O-(C 4 alkyl), C 4 alkyl is not equal to tert-butyl; and in those pyrrolidine structures (1 is 1; m is 2; n is 0; o is 0; p is 1; X is
CH
2 W is equal to CH,.
In yet another embodiment of the compound of formula R, may occur one to four times and each occurrence is independently hydrogen or C,.
6 alkyl, and R 2 is hydrogen, pyridyl, alkyl, alkyl)-CO2-R, or Additionally, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: WO 99/67230 WO 9967230PCT/US99/1 4233 -16-
/C(CH
2 \NRI I together with the carbon atom to which they are attached. Additionally, in this embodiment, R 3 is hydrogen, phenyl, C,.
6 alkyl, C 3 6 alkenyl, C,- 1 arylalkyl (C 6 alkyl)-C0 2 1
(C
2 6 alkenyl)-C0 2 alkyl)-CO-C,.
6 alkyl,
(C
1 6 alkyl)-O-C,-6alkyl, (CI.
6 alkyl)-OH, (C, 4 alkyl)-CN, -adamantyl or one of the following: 16 i
N
H
16 alkyI)-N Z N4-J q -Nq
N
-0-~CO--RI
I
CO,.R
N
-CG-C0 2
-R
1 6 alkyl)-:-
N
3 aflyI)], 3 GC02-Rjj
N
OH
0 -0 -CF 3
S
H C0 2 -RI I <Nli H CO,-R,,I 0 NO, HN-N 1' C0 2 -Rjj
N(CI-
3 alkYI), In addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: 100 Additionally, R4 is 1, NH 2 NHOH, -0-(C 7 -10 arylalkyl), or is of the formula WO 99/67230 PCT/US99/14233 -17- -0-CH2-fN Also, in this embodiment, R, is a formula of the following:
RS
R8-R12 R7 R Additionally, in this embodiment, R 6 is N or CH, R, is hydrogen or halogen, R 8 is -NHCO-, R, may occur one to three times and is a halogen, Ci.
6 alkoxy, C,.
6 alkyl or trifluoromethyl, RIo is CI.
6 alkyl, or alkyl)-OH, or hydrogen, is hydrogen or C, 6 alkyl,R,2 is C,.
6 alkyl or the following formula: Rl3
-(CH
2 R9 R3 is N or CH, W is (C, 6 alkyl), X is S, O, or CH 2 Y is or -C(=O)N(Ri 0 Z is O, CH 2 or is 1, 2, or 3, m is 1 or 2, n is 0 or 1, o is 0 or 1, p is 0 or 1, q is 0 or 1, and r is 0, 1, 2 or 3. This particular embodiment of the present invention has the provisos that when Y is R 3 cannot be hydrogen; when R 4 is equal to O-(C 4 alkyl), C 4 alkyl is not equal to tert-butyl; in those pyrrolidine structures (1 is 1; m is 2; n is 0; o is 0; p is 1; X is
CH
2 W is equal to CH 2 and the compound has an IC 5 0 value of less than 51pM in a Jurkat CS-1 assay and/or an ICo value of less than 50pM in a Jurkat EC assay.
In another embodiment of the compound of formula R, may occur one to four times and each occurrence is independently hydrogen or C 1 alkyl, R, is hydrogen, pyridyl, C_ 6 alkyl, 6 alkyl)-CO2-RI,, or Additionally, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: WO 99/67230 PCTIUS99/14233 -18- C\ (CH 2 )1\NRII C NRI 1 (CH2) together with the carbon atom to which they are attached. Also, in this embodiment, R 3 is hydrogen, phenyl, C, 6 alkyl, C 36 alkenyl, arylalkyl, (C, 6 alkyl)-C 2
(C
2 6 alkenyl)-C 2
(C
6 alkyl)-CO-C,.
6 alkyl, 6 alkyl)-O-C,- alkyl, alkyl)-OH, 6 alkyl)-CN, adamantyl or one of the following: ~c lI yORjj NN
H
1-6 alkyl-N Z q -N Z
N
CO)-RI
[O(C-3alkyl)]-3 CO)2-RI I
NN
-2-R (C-6a
N
N
-G
CO
2
-R
1 1
OH
0 _0 N CF3 S C2-R H C0 2
I
H CO2-R I
NO,
HN-N
-JZ C0-Rj, J(CI-3alyl)2 In addition, R, and R 3 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N In this embodiment, R 4 is NH,, NHOH, arylalkyl), or R 4 is of the formula: O-CH24 WO 99/67230 PCT[US99/14233 -19- In this embodiment, R, is a formula of the following:
R
8
-R
1 2 Also, in this particular embodiment, R~is N or CH, R 7 is hydrogen or halogen, R, is
-OCH
2
R
9 may occur one to three times and is a halogen, C 14 alkoxy,C 6 alkyl or trifluoromethyl, Rio is C,.
6 alkyl, (C 1 6 alkyl)-OH, or hydrogen, is hydrogen orCalkyl, R, 2 is C~ 6alkyl or the following formula: -(CH2)r R R 3 is N or CH, W is (C 6 alkyl), X is S, 0, or CH 2 Y is -CO-, -C(=0)N(R 0 Z is 0, CH 2 or N-R 1 1 is 1, 2, or 3, m is 1 or 2, n is 0 or 1, o is 0or 1, pis 0or 1, qis 0or 1, and ris 0, 1, 2 or 3. This particular embodiment has the following provisos: when Y is R 3 cannot be hydrogen; when R 4 is equal to 0-(C 4 alkyl), C 4 alkyl is not equal to tert-butyl; in those pyrrolidine structures (I is 1; m is 2; n is 0; o is 0; p is 1; X is W is equal to CH 2 when R 3 is phenyl, C 1 6 alkyl, C 7 1 7 arylalkyl, (C 1 6 alkyl)-C0 2 -R 1
(C,
4 alkyl)-O- C,.
6 alkyl or (C 1 6 alkyl)-OH, o is 0; and the compound is not Cl H o C02H N C WO 99/67230 WO 9967230PCTIUS99/I 4233 In yet another embodiment of the compound of formula R, may occur one to four times and each occurrence is independently hydrogen or C,- 6 alkyl, and R 2 is hydrogen, pyridyl. C alkyl, (C 1 6 alkyl)-C0 2 or -C0 2 Additionally, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: together with the carbon atom to which they are attached. Additionally, in this embodiment, R.
3 is hydrogen, phenyl, alkyl, C 3 6 alkenyl, arylalkyl (C -6 alkyl)-C0 2
(C
2 6 alkenyl)-C0 2 (C -6 alkyl)-CO-C 6 alkyl, alkyl)-O-C,- 6 alkyl, (C 1 6 alkyl)-OH, 6 alkyl)-CN, adamantyl or one of the following: -(C6alky)---I
H
-(CI1-6 alkyl)--N z -N Z
N
-0-CO 2
R,,
CO'
2 .R I,
N
N
[O(C1; 3 akyI)].
3 GC02-RjI
N
OH
0 fij 5% CF3
S
H C0 2 -Rl I -<01 H C0 2 -Rjj 0 NO-) I-N-N N C0 1
N(C,-
3 alkyI2 In addition, R, and R 3 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: WO 99/67230 WO 9967230PCT/US99/I 4233 -21- Z N,.
0 Additionally, R 4 is NH,, NHOH, 10 arylalkyl), or is of the formula Also, in this embodiment, R, is a formula of the following:
R
6 Additionally, in this embodiment, R 6 is N or CH, R 7 is hydrogen or halogen, R 8 is -0CH 2 ,1
R
9 may occur one to three times and is a halogen, alkoxy, alkyl or trifluoromethyl,
R
10 iS C1.
6 alkyl, or 6 alkyl)-OH, or hydrogen, is hydrogen or C,.
6 alkyl, R 1 2 is C 1 -6 alkyl or the following formula:
R
13 -(CH2)r R R 3 is N or CH, W is (C alkyl), X is S, 0, or CH, Y is -CO-, -C(=0)N(R 0 Z is 0, CH 2 or 1 is 1, 2, or 3, m is 1 or 2, n is 0or 1, o isO0 or l, p isO0 or 1, q isO0 or 1, and r isO0, 1, 2 or 3. This particular embodiment of the present invention has the provisos that: when Y is R 3 cannot be hydrogen; when 1R4 is equal to 0-(C 4 alkyl), C 4 alkyl is not equal to tert-butyl; in those pyrrolidine structures (I is 1; m is 2; n is 0; o is 0; p is 1; X is CHA) W is equal to CH 2 the compound has an IC 50 value of less than 5 tM in a Jurkat CS-I assay and/or an IC, 0 value of less than 50OpM in a Jurkat EC assay; when R(3 is phenyl, C 1 -6 alkyl, C 71 arylalkyl, alkyl)-C0 2 WO 99/67230 PCT/US99/14233 -22- 6 alkyl)-O- C,.
6 alkyl or alkyl)-OH, o is 0; and the compound is not Cl H 0 CO 2
H
Os
H
In the above formula the absolute stereochemistry for the bond leading to R, is shown, however the absolute stereochemistry has not been shown for all examples which follow. It is understood that all other formulas also follow this type of absolute stereochemistry unless otherwise stated. Additionally, it is to be understood by those of skill in the art that the present invention embodies stereochemical configurations other than those shown. Specifically, the present invention embodies all configurations including the various stereoisomers. Compounds which do not meet the absolute stereochemistry in formula (I) should meet an activity threshold in various assays, to be explained below, which can ensure their efficacy as useful molecules. The present invention includes mixtures, such as racemic mixtures, which contain molecules having the claimed stereochemistry.
The desired compound of the present invention may be clinically used either in a free form or in the form of pharmaceutically acceptable salts thereof. Pharmaceutically acceptable salts include acid-addition salts with inorganic acid or organic acid hydrochloride, sulfate, nitrate, hydrobromide, methanesulfonate, p-toluenesulfonate, acetate), salt with inorganic base, organic base or amino acid triethylamine salt, a salt with lysine, an alkali metal salt, an alkali earth metal salt and the like).
The compound may also be formulated into a pharmaceutical composition comprising a therapeutically effective amount of the compound as defined above and a pharmaceutically acceptable carrier or diluent.
The compound can also be used for treating or preventing a4fP adhesion mediated conditions in a mammal such as a human. This method may comprise administering to a WO 99/67230 PCT/US99/14233 -23mammal or a human patient an effective amount of the compound or composition as explained above.
This method can be used to treat such inflammatory conditions as rheumatoid arthritis, asthma, allergy conditions, adult respiratory distress syndrome,
AIDS,
cardiovascular diseases, thrombosis or harmful platelet aggregation, reocclusion following thrombolysis, allograft rejection, reperfusion injury, psoriasis, eczema, contact dermatitis and other skin inflammatory diseases, osteoporosis, osteoarthritis, atherosclerosis. neoplastic diseases including metastasis of neoplastic or cancerous growth, wound healing enhancement, treatment of certain eye diseases such as detaching retina, Type I diabetes, multiple sclerosis, systemic lupus erythematosus (SLE), inflammatory and .immunoinflammatory conditions including ophthalmic inflammatory conditions and inflammatory bowel diseases, ulcerative colitis, atherosclerosis, regional enteritis and other autoimmune diseases.
As mentioned above, the compounds and compositions containing the compounds according to the present invention are particularly useful in treating or preventing acP adhesion mediated conditions in a mammal such as a human. The present inventors have found that the compounds and compositions containing the compounds according to the present invention are most useful in the treatment of asthma.
The desired compound of the present invention or pharmaceutically acceptable salts 20 thereof may be administered either orally or parenterally, and it may be used as a suitable pharmaceutical preparation, for example, a tablet, a granule, a capsule, a powder, an injection, and an inhalation by a conventional process.
The dose of the desired compound of the present invention or a pharmaceutically acceptable salt thereof varies depending on an administration method, age, body weight, and state of a patient, but, in general, the daily dose is preferably about 0.1 to 100 mg/kg/day, however, I to 100 mg/kg/day may also be suitable.
Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.
WO 99/67230 PCT/US99/1 4233 -24- Preferred routes of administration for asthma: It is preferred that the compound of the present invention be administered in the form of an Aerosol. However, other routes of administration include intravenous, oral, intramuscular, and subcutaneous.
In the case of aerosol administration, compositions containing the compounds of the present invention can be prepared to provide for an excellent means for administering in aerosol form for inhalation therapy. Accordingly, the present invention will provide for selfpropelling compositions containing the compounds of the present invention.
Propellants employed should be non-toxic and have a vapor pressure suitable for the conditions under which administration occurs. These propellants can be fluorinated or fluorochlorinated lower saturated aliphatic hydrocarbons. The preferred propellants of this type are the halogenated alkanes containing not more than two carbon atoms and at least one fluorine atom. Illustrative of these are trichloromonofluoromethane, dichlorodifluoromethane, monochlorotrifluoromethane, dichloromonofluoromethane and 1,2-dichloro-l,l,2,2-tetrafluoroethane. These compounds are available from E.I. duPont de Nemours and Company under the trade name "Freon". These propellants may be employed singularly or in admixture.
In addition to the propellant, an organic solvent may also be employed. The organic solvent must be non-toxic and without undesirable effects on inhalation in the amount present in the aerosol produced. In addition, the solvent should be substantially anhydrous, completely miscible with the propellant or mixture of propellants employed and have a suitable boiling point. Examples of such solvents included non-toxic aliphatic alcohols such as ethanol; ethers such as ethyl ether and vinyl ether; ketones such as acetone; and suitable halogenated lower alkanes.
In addition to the organic solvent, the composition may also optionally contain a nontoxic hygroscopic glycol. The glycol must be substantially miscible with the organic solvent and the propellant employed. Satisfactory glycols include propylene glycol, triethylene glycol, glycerol, butylene glycol and hexylene glycol.
The above indicated methods of administration and formulation of aerosol compositions should not be viewed as limiting. The compounds of the present invention can WO 99/67230 PCT/US99/14233 be formulated in anyway deemed suitable to one of ordinary skill in the art so as to obtain the desired effects.
Pharmaceutical Compositions As indicated previously, the compounds of formula can be formulated into pharmaceutical compositions. In determining when a compound of formula is indicated for the treatment of a given disease, the particular disease in question, its severity, as well as the age, sex, weight, and condition of the subject to be treated, must be taken intd6 consideration and this perusal is to be determined by the skill of the attendant physician.
For medical use, the amount of a compound of formula required to achieve a therapeutic effect will, of course, vary both with the particular compound, the route of administration, the patient under treatment, and the particular disorder or disease being treated. A suitable daily dose of a compound of Formula or a pharmaceutically acceptable salt thereof, for a mammalian subject suffering from, or likely to suffer from, any condition as described hereinbefore is 0.1 mg to 100 mg of the compound of formula I, per kilogram body weight of the mammalian subject. In the case of systematic administration, the dose may be in the range of 0.5 to 500 mg of the compound per kilogram body weight, the most preferred dosage being 0.5 to 50 mg/kg of mammal body weight administered two to three times daily. In the case of topical administration, to the skin or eye, a suitable dose may be in the range of 0.1 jig to 100 jig of the compound per kilogram, typically about 0.1 gjg/kg.
In the case of oral dosing, a suitable dose of a compound of Formula or a physiologically acceptable salt thereof, may be as specified in the preceding paragraph, but most preferably is from 1 mg to 10 mg of the compound per kilogram, the most preferred dosage being from 1 mg to 5 mg/kg of mammal body weight, for example, from 1 to 2 mg/kg. Most preferably, a unit dosage of an orally administrable composition encompassed by the present invention contains less than about 1.0 g of a formula compound.
It is understood that formulation, both for human and veterinary use, of the present invention may be presented to the mammal by inhalation. To achieve therapeutic effect, the dose may be in the range of 0.5 to 500 mg of the compound, per kg body weight. The most WO 99/67230 PCT/US99/14233 -26preferred dosage being 0.5 to 50 mg/kg of mammal body weight administered two to three times daily.
It is understood that the ordinarily skilled physician or veterinarian will readily determine and prescribe the effective amount of a compound of formula to prevent or arrest the progress of the condition for which treatment is administered. In so proceeding, the physician or veterinarian could employ relatively low doses at first, subsequently increasing the dose until a maximum response is obtained.
The compounds and compositions of the present invention can be administered to patients suffering from a condition listed herein in an amount which is effective to fully or partially alleviate undesired symptoms of the condition. The symptoms may be caused by inappropriate cell adhesion mediated by a43 1 integrins. Such inappropriate cell adhesion would typically be expected to occur as a result of increased VCAM-1 and/or CS-I expression on the surface of endothelial cells. Increased VCAM-I and/or CS-I expression can be due to a normal inflammation response or due to abnormal inflammatory states. In either case, an effective dose of a compound of the invention may reduce the increased cell adhesion due to increased VCAM-1 expression by endothelial cells. Reducing the adhesion observed in the disease state by 50% can be considered an effective reduction in adhesion.
More preferably, a reduction in adhesion by 90%. is achieved. Most preferably adhesion mediated by VCAM-1/c 1 P, and/or CS-I interaction is abolished by an effective dose.
Clinically, in some instances, effect of the compound can be observed or a decrease in white cell infiltration into tissues or a site of injury. To achieve a therapeutic effect, then, the compounds or compositions of the present invention are administered to provide a dose effective to reduce or eliminate inappropriate cell adhesion or to alleviate undesired symptoms.
While it is possible for an active ingredient to be administered alone, it is preferable to present it as a pharmaceutical formulation comprising a compound of formula and a pharmaceutically acceptable carrier thereof. Such formulations constitute a further feature of the present invention.
The formulations, both for human and veterinary medical use, of the present invention comprise an active ingredient of formula in association with a pharmaceutically WO 99/67230 PCT/US99/14233 -27acceptable carrier thereof and optionally other therapeutic ingredient(s), which are generally known to be effective in treating the disease or condition encountered. The carrier(s) must be "acceptable" in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient thereof The formulations include those in a form suitable for oral, pulmonary, ophthalmic, rectal, parenteral (including subcutaneous, intramuscular, and intravenous), intra-articular, topical, nasal inhalation with an aerosol) or buccal administration. Such formulation are understood to include long-acting formulations known in the art.
The formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods may include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing the active ingredient into association with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, shaping the product into the desired form.
Formulations of the present invention suitable for oral administration may be in the form of discrete units such as capsules, cachets, tablets, or lozenges, each containing a predetermined amount of the active ingredient in the form of a powder or granules; in the form of a solution or suspension in an aqueous liquid. Formulations for other uses could involve a nonaqueous liquid; in the form of an oil-in-water emulsion or a water-in-oil emulsion; in the form of an aerosol; or in the form of a cream or ointment or impregnated into a transdermal patch for use in administering the active ingredient transdermally, to a patient in need thereof. The active ingredient of the present inventive compositions may also be administered to a patient in need thereof in the form of a bolus, electuary, or paste.
The practitioner is referred to "Remington: The Science and Practice of Pharmacy," 19th Edition, c. 1995 by the Philadelphia College of Pharmacy and Science, as a comprehensive tome on pharmaceutical preparations.
WO 99/67230 WO 9967230PCTIUS99/I 4233 -28- Abbreviations
AC
2
O:
EtOAc:
BCECF-AM:
B OP-Cl: BOP Reagent:
DMEM:
DMF:
DIEA:
EDC:
Et: EtOH:
HATU:
HBSS:
HBTU:
HOBT (HOBt):
HSA:
LDA:
Me: meq: MeOH: n-Bu:
NMP:
PBS:
Pd-C: Ph: Acetic anhydride Ethyl acetate 2',7'-bis-(2-carboxyethyl)-5 -(and carboxyfluorescein acetoxymethyl ester Bis 2 -oxo-3-oxazolidinyl) phosphinic chloride Benzotriazol-1I-yloxy-tris (dimethylamino)-phosphonium hexafluorophosphate Dulbecco's Minimal Eagle's Media Dimethyl formamide Diisopropylethylamnine 1 3 -Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride Ethyl Ethanol N-[(Dimethylamino)- 1H-I ,2,3-triazolo[4,5-b]-pyridin-
I-
ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide Hank's Balanced Salt Solution O-Benzotriazol- I -yl-N,NN',N'-tetramethyluronium hexafluorophosphate 1 -Hydroxybenzotriazole Human serum albumin Lithium diisopropylamide Methyl milliequivalent Methanol n-Butyl 1 -Methyl-2-pyrrolidinone Phosphate buffered saline Palladium on charcoal Phenyl WO 99/67230 WO 9967230PCTJUS99/1 4233 -29- SPDP: 3 -(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester t-Bu: t-butyl THF: Tetrahydrofuran TEA: Trifluoroacetic acid DMSO: dimethyl sulfoxide HOAt: 1 -hydroxy-7-azabenzotriazole DMAP: 4-dimethylaminopyridine FMOC: 9 -fluorenylmethoxycarbonyl Bn: benzyl PyBOP: (benzotriazol-l-yloxy)tripyrrolidinophosphonium hexafluo4 phosphate BOC: tert-butoxycarbonyl The representative compounds according to the present invention are prepared as described below. The compounds of the present invention are prepared in a similar manner.
WO 99/67230 WO 9967230PCTIUS99/1 4233 Scheme A HS RA-l R2J A2 H A-i 0 R1 SRA2 Nt OH A-2 H 0 S-RA.2 r
OHA-
R,
H
2 N Ol A-4 0 RA.1 RA-1 S RA- 0 S RA.-2 0 y NHJL, 4 NH U,,NO
R
3 -0'LO 0 KR5 A-5
R
3 -0-O 0 R A-9 RA-1 RA-1 S RA- 0
.R
IyNH 0 4 N N R H A-1 H 0 R 0 R-6 RA-1 S 0 I A-7 RA-1 S; RA- 0 4 NH~kO F;0 WO 99/67230 PCT/US99/14233 -31- RA-. and RA.
2 are defined independently as R,.
Scheme A describes a general method for the preparation of examples of the formula A-6, A-7, A-8, A-9 and A-10. A commercially available or readily prepared sulfur containing amino acid of structure A-1 (for the synthesis of P,P-disubstituted cysteine amino acids see: Stanfield, Hruby, V.J. Synth. Commun. 1988, 18, 531 and references therein) is condensed with formaldehyde to afford the thiazolidine-4-carboxylic acid of general formula A-2 (for the condensation of aldehydes with cysteine or similar sulfur containing amino acids see for example: Ratner, Clark, H.T. J. Am. Chem.
Soc. 1937, 59, 200. Lewis, Inloes, Hes, Matthews, Milo, G. J. Med.
Chem. 1978, 21, 1070. Oya, Baba, Kato, Kawashima, Watanbe, T.
Chem. Pharm. Bull. 1982, 30, 440.). Standard protection affords carbamate A-3 which is readily condensed with amino acid derivative A-4 under standard peptide synthesis conditions to provide the psuedodipeptide A-5 (for a review of procedures of peptide synthesis see: Bodansky, Bodansky, A. The Practice of Peptide Synthesis; Springer- Verlag: Berlin, 1984). Deprotection of the carbamate from A-5 provides the useful intermediate A-6. The amine group may be reacted with a variety of electrophilic reagents such as: commercially available or readily prepared sulfonyl chlorides (for the synthesis of sulfonyl chlorides see for examples: Roblin, Clapp, J.W. J. Am.
Chem. Soc. 1950, 72, 4890. Gilbert, E.E. in Sulfonation and Related Reactions Olah, Ed. John Wiley and Sons, New York; 1965. Park, Shin, Kin, Y.H.
Chem Lett. 1992, 1483. Kim, Ko, Kim, S.H. Synthesis, 1992, 1203.) to afford sulfonamides of general structure A-7 where Y is SO 2 (preparation carbonates or chloroformates to afford carbamates of general structure A-7 where Y is CO 2 (preparations 2, 7, isocyanates to afford ureas of general structure A-7 where Y is
CONHR
3 (preparation phosgene or a suitable equivalent and an amine to afford ureas of general structure A-7 where Y is CON(C,.
6 alkyl)R 3 (preparation 10, also see for example: Nowick, Homes, Noronha, Smith, Tram, Huang, S.
J. Org. Chem. 1996, 61, 3929.); acid chlorides and carboxylic anhydrides to provide amides of structure A-7 where Y is CO- (preparation 11). Mild base hydrolysis of monoesters of general structure A-7 (preparation 6) or diesters of general structure A-7 WO 99/67230 PCT/US99/14233 -32- (preparation 12) affords the acids of general structure A-8. Mild base hydrolysis of the ester of general structure A-5 provides acid A-9 (preparation 6 or 13) which may be further deprotected to afford the amino acid A-10 (preparation 14).
Preparation 1 (Scheme A, A-2: where RA.i and RA.
2 are the same and equal to H and stereochemistry is t(N O H H 0 D-Cysteine hydrochloride monohydrate where RA., and RA 2 are the same and equal to H and stereochemistry is (35.04 g, 0.19 mol) was dissolved in formaldehyde wt% solution in water, 38 mL) and the reaction mixture allowed to stir for 18 h at ambient temperature. The mixture was cooled (0-5C and absolute ethanol (93 mL) and pyridine (57 mL) were added. After one hour, the precipitate was collected by filtration, washed with cold absolute ethanol followed by diethyl ether and dried in vacuo to afford the title compound (24.6 g) as a white crystalline solid: mp 181-184 C (Lit. 194-196 Lewis, Inoles, Hes, J. J. Med. Chem. 1978, 21, 1070.); 'H NMR (DMSO-d 6 8 4.22 (1 4.04 (1 3.86 (1 3.09 (1 2.24 (1 MS (ESI+) for C 4 HNO,S m/z 134.0 Preparation 2 (Scheme A, A-3: where RA-I and RA 2 are the same and equal to H, R3 is t-butyl and stereochemistry is s- N0" x[ o A solution of A-2 (Scheme A where RA. and RA.
2 are the same and equal to H and stereochemistry is (24.6 g, 0.185 mol) and di-t-butyl dicarbonate (44.4 g, 0.2 mol) in THF (1 L) was heated to reflux for 18 h. Volatiles were removed in vacuo and the residue WO 99/67230 PCT/US99/14233 -33partitioned between ethyl acetate and 0.1 N NaOH. The aqueous layer was washed with ethyl acetate, made acidic with 1.0 N HCI (pH 3-4) and then extracted with ethyl acetate.
The combined organic extracts were washed with brine, dried (Na 2
SO
4 filtered and concentrated in vacuo. Crystallization of the white solid from hexane/methylene chloride provided the title compound (31.8 g) as white crystals: mp 132-134 OC; [a] 2 5 D 1170 (c 0.66, ethanol); IR (mull) 3002, 1747, 1635, 1421, 1404, 1393, 1310, 1215, 1198, 1166, 1144, 1122, 894, 862, 774 'H NMR (DMSO-d 6 6 4.57 (2 4.28 (1 3.09 (1 H), 1.35 (9 "C NMR (DMSO-d 6 8 171.8, 152.5, 79.8, 60.9, 48.4, 47.7, 33.8, 32.6, 27.7; MS (ESI+) for C 9
H,,NO
4 S m/z 234.2 MS (ESI-) for C 9
H,
5
NO
4 S m/z 232.1 Anal. Calcd for C 9 H,,NO4S: C, 46.34; H, 6.48; N, 6.00. Found: C, 46.27; H, 6.48; N, 6.03.
Preparation 3, Example 1 4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(1,1-dimethylethyl) ester (Scheme A, A-5: where RA., and RA.
2 are the same and equal to H, R 3 is t-butyl, R 5 is 4- 2 6 -dichlorobenzoyl)amino]phenyl, and stereochemistry is
O
N 0 O o0- o To a cooled (0-5°C suspension of A-3 (Scheme A where RA^. and RA2 are the same and equal to H, R 3 is t-butyl and stereochemistry is (8.67 g, 37.2 mmol) and HOBT (5.69 g, 37.2 mmol) in CH 2 C1 2 (60 mL) was added a solution of EDC (7.12 g, 37.2 mmol) in
CH
2
CI
2 (140 mL). After 30 min at 0-5 0 C A-4 (where R 5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is (10 g, 24.8 mmol) was added followed by 4methylmorpholine (2.72 mL, 24.8 mmol). The reaction mixture was gradually warmed to ambient temperature, stirred an additional 18 h and diluted with CH 2 C1 2 and 0.1 N HC1.
The organic layer was separated and washed with 0.1 N HC1, sat. aqueous NaHCO 3 brine, WO 99/67230 PCT/US99/14233 -34dried (NaSO 4 filtered and concentrated in vacuo. Flash chromatography of the residue using hexane/ethyl acetate as eluant afforded the title compound (13.9 g) as a white solid. Recrystallization from acetone/hexane afforded a crystalline solid: mp 222-224°C; IR (mull) 3282, 3254, 1738, 1714, 1707, 1678, 1662, 1610, 1562, 1545, 1431, 1414, 1287, 1256, 784 cm'; 'H NMR (CDC1 3 8 7.57 (2 7.34 (5 7.14 (2 4.74 (3 4.30 (1 3.74 (3 3.37 (1 3.15 (3 1.45 (9 "C NMR (DMSO-d 6 8 172.1, 162.3, 153.2, 137.6, 136.9, 133.2, 131.8, 131.7, 130.1, 128.7, 119.8, 80.4, 62.2, 53.7, 52.4, 36.7, 28.3; MS (ESI+) for C 26
H
29
CI
2
N
3 06S m/z 604.3 Anal. Calcd for C2 6
H
2 9
CI
3
N
3 0 6
S:
C, 53.61; H, 5.02; N, 7.21. Found C, 53.82; H, 4.81; N, 7.22.
Preparation 4 (Scheme A, A-6: where RA., and RA.
2 are the same and equal to H, R 5 is dichlorobenzoyl)amino]phenyl, and stereochemistry is S- O H 0 H To a cooled (5-10°C solution of A-5 (Scheme A where RA., and RA2 are the same and equal to H, R 3 is t-butyl, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl, and stereochemistry is (3.3 g, 5.67 mmol) in dioxane (34 mL) was added a solution of HCl in dioxane (4 M, 140 mL) in a dropwise manner over 30 min. After an additional 30 min at 0-5 0 C the ice bath was removed and the reaction mixture stirred 1 h at ambient temperature. The volatiles were removed in vacuo to afford the title compound (2.94 g) as a light yellow solid: 'H NMR (DMSO-d 6 8 10.70 (1 9.1 (1 7.52 (5 7.18 (2 4.64 (1 H), 4.37 (1 4.21 (2 3.67 (3 3.10 (1 2.89 (1 2.70 (1 "C NMR (DMSOd 6 6 172.1, 162.3, 153.2, 137.6, 136.9, 133.2, 131.8, 131.7, 130.1, 128.7, 119.8, 80.4, 62.2, 53.7, 52.4, 36.7, 28.3; MS (ESI+) for C2,H 2 ,C1 2
N
3 0 4 S m/z 482.1 WO 99/67230 PCT/US99/14233 Preparation 5 and Example 2.
2 6 -Dichlorobenzoyl)amino]-N-[[(4S)-3-(methylsulfonyl)-4 thiazolidinyl]carbonyl]-L-phenylalanine methyl ester (Scheme A, A-7: where RA.- and RA2 are the same and equal to H, R 3 is methyl, Y is SO2,, R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemistry is
O
I ~~QNJ To a cooled (0-5 0 C) solution of A-6 (Scheme A, where RA-1 and RA.
2 are the same and equal to H, Rs is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is g, 3.86 mmol) in anhydrous THF (50 mL) was added methanesulfonyl chloride (2.99 mL, 38.6 mmol) followed by pyridine (6.24 mL, 77.2 mmol). After 1 h at 0-5 0 C the ice bath was removed and the solution stirred at ambient temperature for 2 h then diluted with ethyl acetate and 0.25 N HCI. The layers were separated and the organic layer washed with sat.
aqueous NaHCO 3 brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using ethyl acetate/methylene chloride/hexane and isopropanol as eluant afforded the title compound (1.99 g) as an amorphous powder: IR(mull) 1742, 1666, 1605, 1562, 1534, 1515, 1432, 1413, 1344, 1327, 1269, 1218, 1195, 1156, 780 cm'; 'H NMR (300 MHz, CDC13) 5 7.55 (3 7.33 (3 7.12 (3 4.84 (1 4.69 (1 4.61 (1 4.29 (1 3.74 (3 3.50 (1 H), 3.29 (1 3.14 (2 2.92 (3 3 C NMR (75 MHz, CDCI 3 6 171.2, 168.1, 162.4, 136.5, 135.9, 132.3, 132.2, 130.8, 130.0, 128.1,120.6, 64.9, 53.2, 52.5, 51.8, 37.2, 34.1; MS (ESI+) for C, 2
H
23
C
2
N
3 0 6
S
2 m/z 559.8 HRMS (FAB) calcd for
C
22
H
23
CL
2
N
3 0 6 S2+H 560.0483, found 560.0504; Anal. Calcd for C 22
H
23 C1 2
N
3 0 6 S2: C, 47.15; H, 4.14; N, 7.50. Found: C, 46.88; H, 4.32; N, 7.16.
WO 99/67230 PCTIUS99/1 4233 -36- Preparation 6 and Example 3.
4 2 ,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-(methylsulfonyl)-4thiazolidinyl]carbonyl]-L-phenylalanine (Scheme A, A-8: where RA., and RA.
2 are the same and equal to H, R 3 is methyl, Y is SO2,
R
5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is N OH O=S=O 0 0 CI To a cooled (0-5 0 C solution of A-7 (Scheme A, where RA., and RA.
2 are the same and equal to H, R 3 is methyl, Y is SO2, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (1.75 g, 3.12 mmol) in THF (100 mL) and water (10 mL) was added an 0.1 N aqueous solution of NaOH (34.3 mL, 3.43 mmol) via a syringe pump over 1 h. After an additional 45 min at 0-5 0 C, the reaction mixture was diluted with ethyl acetate and acidified with 0.25 N HCI to a pH of ca. 3. The organic layer was separated, washed with water and concentrated in vacuo. Purification of the residue by flash chromatography using methylene chloride and methanol as eluant provided a solid which was lyophilized from glacial acetic acid to provide the title compound (1.42g) as an amorphous powder: 25 D 103°(c 0.97, ethanol); IR (mull) 3291, 1736, 1666, 1605, 1562, 1534, 1516, 1432, 1414, 1339, 1270, 1195, 1154, 799, 780cm-'. 'HNMR (300 MHz, CD 3 OD) 5 7.58 (2 7.45 (3 7.25 (2 4.72 (2 4.37 (1 3.17 2.99 (3 "C NMR (75 MHz, CD 3 OD) 8 174.1, 171.6, 165.2, 138.3, 137.7, 134.8, 133.4, 132.4, 131.2, 129.4, 121.6, 66.2, 55.0, 52.9, 37.9, 37.5, 35.7; MS (ESI+) for
C
2
,H
2 ,C1 2
N
3 0 6
S
2 m/z 545.8 HRMS (FAB) calcd for C,IH 2
,CL
2
N
3 0 6
S
2
+H,
546.0327, found 546.0358. Anal. Calcd for C 21
H
2 C1l 2
N
3 0 6 C, 46.16; H, 3.87; N, 7.69.
Found: C, 46.24; H, 4.04; N, 7.33.
WO 99/67230 PCT/US99/14233 -37- Preparation 7 and Example 4.
4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester (Scheme A, A-7: where RA.I and RA_ 2 are the same and equal to H, R 3 is ethyl, Y is CO2-, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is H 0 N O 00
CI
To a cooled (0-5 0 C solution of A-6 (where R^A- and RA 2 are the same and equal to H, R is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is (1.25 g, 2.40 mmol) in anhydrous THF (30 mL) was added ethyl chloroformate (340 pL, 3.60 mmol) followed by triethylamine (810 gL, 5.79 mmol). After 1 h at 0-5C the ice bath was removed and the solution stirred at ambient temperature for 2 h then diluted with ethyl acetate and 0.25 N HC1. The layers were separated and the organic layer washed with sat.
aqueous NaHCO 3 brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using ethyl acetate/methylene chloride/hexane and isopropanol as eluant afforded the title compound (1.10 g) as an amorphous powder: 'H NMR (300 MHz, CDC1 3 8 7.59 (2 7.30 (3 7.10 (2 H), 4.81 (1 4.72 (2 4.38 (1 4.11 (2 3.19 (4 1.25 (3 "C NMR (75 MHz, CDCl 3 8 171.6, 163.0, 136.9, 136.2, 132.3, 132.1, 130.7, 129.8, 128.0, 120.3, 63.0, 62.8, 57.1, 53.2, 52.5, 37.0, 14.4; MS (ESI+) for C 24
H
25
CI
2
N
3 0 6 S m/z 554.2 MS (FAB) m/z (rel. intensity) 554 (MH, 99), 557 556 555 554 349 245 175 173 160 88 HRMS (FAB) calcd for
C
24
H
2 5
CL
2
N
3 0 6 S 554.0919, found 554.0908. Anal. Calcd for C 24
H
25 Cl 2
N
3 0,S: C, 51.99; H, 4.55; N, 7.58.
Found: C, 52.05; H, 4.67; N, 7.44.
WO 99/67230 PCT/US99/14233 -38- Preparation 8 and Example 4 2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[2-(1-piperidinyl)ethyl] ester (Scheme A, A-7: where RA.- and RA.2 are the same and equal to H, R 3 is 2-(1piperidinyl)ethyl, Y is CO 2 R, is 4 -[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is S- O 0 o c The title compound was prepared by a modification of the literature procedure of Ghosh, Duong, McKee, Thompson, W.J. Tetrahedron Lett. 1992, 33, 2781. To a solution of 1-(2-hydroxyethyl)piperidine (5.11 g, 39.6 mmol) in CH 3 CN (220 mL) at ambient temperature was added N,N-disuccinimidyl carbonate (10.13 g, 39.6 mmol) and triethylamine (16.6 mL, 118.8 mmol). The solution was stirred at room temperature for 4 h and concentrated in vacuo to give a viscous oil. The oil was dissolved in a minimal amount of methylene chloride (50 mL) and added to a solution of A-6 (Scheme A, where RA-I and RA 2 are the same and equal to H, R 5 is 4 2 ,6-dichlorophenyl)methoxy]phenyl and stereochemistry is (2.0 g, 3.96 mmol), triethylamine (0.60 mL) and DMAP mg) in CH 2 Cl 2 (10 mL). The solution was stirred overnight and an additional 5 equivalents of carbonate in methylene chloride (10 mL) [prepared as described above from N,Ndisuccinimidyl carbonate (5.6 g, 19.8 mmol), triethylamine (8.3 mL, 59.4 mmol), and 1- (2-hydroxyethyl)piperidine (2.56 g, 19.8 mmol)] were added. After 3 h at room temperature, propylamine (30 mL, 0.71 mol) was slowly added (exothermic) and the solution diluted with CH 2 C1 2 The resulting solution was stirred vigorously for 15 min.
and diluted with water. The organic layer was separated and washed with 0.1 M HC1, sat.
aqueous NaHCO 3 dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using ethyl acetate/methylene chloride as eluant WO 99/67230 PCT/US99/14233 -39followed by trituration in hexanes afforded the title compound (1.54 g, 62%) as an white powder: IR (mull) 1745, 1704, 1660, 1553, 1512, 1435, 1426, 1397, 1303, 1245, 1227, 1212, 1173, 1019, 765 cm 'H NMR (300 MHz, DMSO-d 6 6 8.42 (1 7.54 (2 H), 7.44 (1 7.13 (2 6.94 (2 5.17 (2 4.59 (2 4.48 (1 4.26 (1 4.02 (2 3.63 (3 3.19 (2 3.19 (1 2.82 (3 2.35 (4 1.36 (6 3 C NMR MHz, DMSO-d 6 5 172.2, 170.0, 157.7, 153.9, 136.5, 132.2, 132.0, 130.8, 130.1, 129.2, 114.8, 65.3, 63.6, 57.3, 54.6, 53.9, 52.4, 36.4, 26.0, 24.3; MS (ESI+) for C 29
H
35 Cl 2
N
3 06S m/z 623.9 Anal. Calcd for C 2 9
H
3 5 Cl 2
N
3 0 6 S: C, 55.77; H, 5.65; N, 6.73. Found: C, 55.48; H, 5.73; N, 6.91.
Preparation 9 and Example 6.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-[(( 1,1 -dimethylethyl)amino)carbonyl]-4thiazolidinyl]carbonyl]-L-phenylalanine methyl ester (Scheme A, A-7: where RA^. and RA.
2 are the same and equal to H, R 3 is t-butyl, Y is CONH-, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is
O
N 0 To a cooled (0-5 0 C solution of A-6 (Scheme A, where R-k. and RA.
2 are the same and equal to H, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is (140 mg, 0.27 mmol) in anhydrous THF (6 mL) was added tert-butyl isocyanate (0.62 mL, 5.4 mmol) followed by 4 -dimethylaminopyridine (5 mg, 0.04 mmol). After 0.5 h at 0-5 0 C, the ice bath was removed and the solution stirred at ambient temperature for 16 h. Additional tert-butyl isocyanate (0.62 mL, 5.4 mmol) was added and the solution warmed to 50 0 C for 4 h. The reaction mixture was cooled to room temperature and diluted with ethyl acetate and 0.25 N HC1. The layers separated and the organic layer washed with sat. aqueous NaHCO 3 brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using ethyl acetate/methylene chloride and WO 99/67230 PCT/US99/14233 isopropanol as eluant afforded the title compound (150 mg) as an amorphous powder: 'H NMR (300 MHz, CDCl 3 6 7.93 (1 7.54 (2 7.26 (4 7.09 (2 4.74 (2 4.66 (1 4.41 (1 4.23 (1 3.70 (3 3.28 (1 3.09 (3 1.31 (9 H); 3 C NMR (75 MHz, CDC1 3 8 171.4, 170.6, 162.4, 155.5, 136.3, 135.9, 132.5, 132.3, 130.8, 129.9, 128.1, 120.6, 62.7, 53.2, 52.4, 51.5, 49.0, 37.0, 32.9, 29.2; MS (ESI+) for
C
26
H
30 C1 2
N
4 0sS m/z 581.0 603.0 MS (FAB) m/z (rel. intensity) 581 23), 482 97 88 83 69 57 55 43 43 (69), 41 HRMS (FAB) calcd for C 26
H
30
CL
2
N
4 0 5 S +H1 581.1392, found 581.1376.
Preparation 10 and Example 7.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-[(diethyamino)carbonyl]-4thiazolidinyl]carbonyl]-L-phenylalanine methyl ester (Scheme A, A-7: where RA^. and RA-2 are the same and equal to H, R, is ethyl, Y is
CON(CH
2
CH
3 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is (S, N0°o 0 0 C1
H
0 CI The title compound was prepared by a modification of the literature procedure of Majer, Randad, R.S. J. Org. Chem. 1994, 59, 1937. A cooled (0-5 0 C) solution of A-6 (Scheme A, where RA^. and RA2 are the same and equal to H, R 5 is dichlorobenzoyl)amino]phenyl and stereochemistry is (200 mg, 0.39 mmol) and triethylamine (56 uL, 0.40 mmol) in anhydrous methylene chloride (10 mL) was added to triphosgene (47 mg, 0.16 mmol) followed by additional triethylamine (56 pL, 0.40 mmol).
After 0.5 h at 0-5 0 C the ice bath was removed and the solution stirred at ambient temperature for 2 h. The solution was re-cooled (0-5C and diethylamine (1.20 mL, 11.70 mmol) and 4 -dimethylaminopyridine (5 mg, 0.04 mmol) were added. After 0.5 h at 0 C the ice bath was removed and the solution stirred at ambient temperature for 16 h.
WO 99/67230 PCT/US99/14233 -41- The solution was concentrated in vacuo and the residue partitioned between ethyl acetate and 0.25 N HC1. The layers were separated and the organic layer washed with sat.
aqueous NaHCO 3 brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using ethyl acetate/methylene chloride/hexane and isopropanol as eluant afforded the title compound (200 mg) as an amorphous solid: 'H NMR (300 MHz, CDC1 3 8 7.86 (1 7.55 (2 7.30 (3 7.03 (2 6.90 (1 5.09 (1 4.83 (1 4.33 (2 3.76 (3 3.34 (3 3.11 (5 H), 1.08 (6 3 C NMR (CDCl 3 8 171.1, 169.6, 162.3, 162.2, 136.5, 135.9, 132.3, 132.1, 130.9, 129.9, 128.1, 120.3, 64.7, 53.4, 52.7, 52.5, 42.0, 37.0, 32.4, 13.1; MS (ESI+) for
C
26
H
30
N
4 0S m/z 580.9 HRMS (El) calcd for C 26
H
3 0
CL
2
N
4 0,S 580.1314, found 580.1297. Anal. Calcd for C 26
H
30 C1lN 4 0sS: C, 53.70; H, 5.20; N, 9.63. Found: C, 53.63; H, 5.33; N, 9.36.
Preparation 11 and Example 8.
4 2 6 -dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-y-oxo-3-thiazolidinebutanoic acid 3-methyl ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to H, R 3 is CHCH 2
CO
2
CH
3 Y is CO, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is s H 0 S 0 o ci 0 NI To a cooled (0-5 0 C solution of A-6 (Scheme A, where RA., and RA 2 are the same and equal to H, R 5 is 4-[(2,6-dichlorobenzoyl)amino] phenyl and stereochemistry is S)) (1.03 g, 1.72 mmol) in anhydrous CH 2 C1 2 (25 mL) was added triethylamine (460 pL, 3.27 mmol) followed by methyl succinyl chloride (320 tL, 2.58 mmol). After 1 h at 0-5 0
C,
the ice bath was removed and the solution stirred at ambient temperature for 2 h then diluted with 1 N HC1. The organic layer was separated, washed with sat. aqueous NaHCO 3 brine, dried (Na 2
SO
4 filtered and concentrated in vacuo. Crystallization of the WO 99/67230 PCT/US99/14233 -42yellow solid from ethanol/water provided the title compound (824 mg) as a light yellow solid: mp 221-223 IR (mull) 3275, 1748, 1731, 1687, 1626, 1610, 1561, 1542, 1517, 1430, 1416, 1326, 1268, 1224, 1193 'H NMR (DMSO-d 6 8 10.64 (1 8.59 (1 H), 8.24 (1 7.50 (5 7.16 (2 4.75 (2 4.51 (2 4.23 (1 3.63 (3 3.56 (3 2.87 (5 "C NMR (DMSO-d 6 172.9, 171.6, 169.9, 169.5, 161.9, 137.2, 136.5, 133.0, 132.8, 131.3, 129.8, 128.3, 119.4, 61.6, 53.6, 52.1, 51.4, 48.6, 36.4, 35.7, 35.2, 33.1, 29.0, 28.9, 28.5; MS (ESI+) for C 2 6
H
27
CI
2
N
3 0 7 S m/z 596.0 MS (ESI-) for
C
26
H
27 C1 2
N
3 0 7 S m/z 593.9 Anal. Calcd for C 26
H
2 7
CI
2
N
3 0 7 S: C, 52.35; H, 4.56; N, 7.04. Found: C, 52.11; H, 4.47; N, 6.96.
Preparation 12 and Example 9.
[S-(R*,R*)]-4-[[[1-Carboxy-2-[4-[(2,6-dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-y-oxo-3-thiazolidinebutanoic acid (Scheme A, A-8: where RA^. and RA.
2 are the same and equal to H, R 3 is
CH
2
CH
2
CO
2
CO
2 H, Y is CO, R 5 is 4 2 ,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is Ho 0N To a cooled (0-5C solution of A-7 (Scheme A where RA^. and RA-2 are the same and equal to H, R 3 is CH2CH 2
CO
2 CH,, Y is CO, R 5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is (130 mg, 0.22 mmol) in anhydrous THF (5 mL) and MeOH (1 mL) was added an aqueous (2 mL) solution of lithium hydroxide monohydrate (23 mg, 0.55 mmol) via a syringe pump over 1 h. After an additional 1 h at 0-5 0 C, the ice bath was removed and the solution stirred 2 h at ambient temperature. The reaction mixture was diluted with ethyl acetate and 0.1 N HCI and the organic layer was separated, washed with water, dried (Na 2
SO
4 filtered and concentrated in vacuo. Lyophilization of the residue from glacial acetic acid afforded the title compound (101 mg) as a white amorphous WO 99/67230 PCT/US99/14233 -43powder: 'H NMR (CD 3 CN) 8 8.87 (1 7.55 (2 7.42 (3 7.22 (2 7.16 (1 H), 4.95 (1 4.52 (3 3.12 (5 2.55 (5 3 C NMR (CD 3 CN) 8 173.8, 171.7, 171.1, 169.8, 162.5, 136.7, 136.0, 133.5, 131.7, 131.3, 130.1, 129.2, 128.2, 119.7, 62.4, 53.6, 48.8, 36.2, 32.4, 29.1, 28.6; MS (ESI-) for C 24
H
23 C1 2
N
3 0,S m/z 566.1 Anal. Calcd for C 24
H
23 C1 2
N
3 0,S: C, 50.31; H, 4.13; N, 7.33. Found: C, 50.13; H, 4.37; N, 6.93.
Preparation 13 and Example [S-(R*,R*)]4-[[[1-Carboxy-2-[4-[(2,6-dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(I,1-dimehylelethyl) ester (Scheme A, A-9: where RA., and RA.
2 are the same and equal to H, R 3 is t-butyl, Y is CO 2
R
5 is 4 2 ,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is s
H
N OH o o ci To a cooled (0-5C of A-5 (Scheme A where R^.A and RA.
2 are the same and equal to H,
R
3 is t-butyl, Y is R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is (150 mg, 0.26 mmol) in anhydrous THF (5 mL) and MeOH (1 mL) was added an aqueous (2 mL) solution of lithium hydroxide monohydrate (14 mg, 0.325 mmol) via syringe pump over 1 h. After an additional 1 h at 0-5 0 C the ice bath was removed and the solution stirred 2 h at ambient temperature. The reaction mixture was diluted with ethyl acetate and 0.1 N HC1, the organic layer separated and washed with water, dried (Na 2
SO
4 filtered and concentrated in vacuo. Lyophilization of the residue from glacial acetic acid afforded the title compound (142 mg) as an amorphous powder: IR (mull) 3285, 1735, 1666, 1606, 1562, 1539, 1516, 1432, 1413, 1394, 1326, 1259, 1219, 1195, 1161 'H NMR (DMF-d 7 5 10.71 (1 8.36 (1 7.91 (2 7.72 (3 7.47 (2 4.88 (3 4.51 (1 3.40 (3 3.22 (2 1.57 (9 "C NMR (DMF-d 7 6 173.3, 163.1, 162.9, 162.7, 162.3, 154.0, 138.3, 137.5, 134.2, 132.3, 131.9, 130.6, 128.9, 120.0, 80.9, 63.1, 54.5, 49.9, 37.4, 28.3; MS (FAB) m/z (rel. intensity) 568 23), WO 99/67230 PCT/US99/14233 -44- 570 568 471 470 65), 469 468 466 175 88 57 HRMS (FAB) calcd for C 25
H
27
CI
2
N
3 06S+H, 568.1075, found 568.1071; MS (ESI-) for C 2 5
H
27 C1 2
N
3 06S m/z 565.8 Anal. Calcd for C 25
H
27
C
2
N
3 0 6 S 0.26 H 2 0: C, 52.38; H, 4.84; N, 7.33. Found: C, 52.07; H, 5.12; N, 7.46; Water 0.83.
Preparation 14 and Example 11.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(4S)-4-thiazolidinyl]carbonyl]-L-phenylalanine monohydrochloride salt (Scheme A, A-10: where RA., and RA.
2 are the same and equal to H, R, is dichlorobenzoyl)amino]phenyl, and stereochemistry is N OH O Co a To a cooled (5-10 0 C solution of A-9 (Scheme A where RA., and RA.
2 are the same and equal to H, R 3 is t-butyl, Y is CO 2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl, and stereochemistry is (193 mg, 0.34 mmol) in dioxane (2 mL) was added a solution of HCI in dioxane (4 M, 8 mL) in a dropwise manner over 30 min. After an additional 3.5 h at 0-5 0 C, the reaction mixture was concentrated in vacuo. Lyophilization of the residue from water afforded the title compound (158 mg) as an amorphous powder: IR (mull) 3248, 3191, 3048, 1731, 1664, 1605, 1577, 1562,1541, 1516, 1431, 1414, 1327, 1195, 799 cm'; 'H NMR (DMF-d,) G 9.11 (1 7.76 (2 7.60 (3 7.35 (2 4.66 (2 H), 4.46 (2 3.55 (3 3.24 (2 3.10 (2 3 C NMR (DMF-d,) 5 172.8, 167.6, 163.1, 162.9, 162.7, 162.3, 138.5, 137.5, 133.8, 132.3, 131.9, 130.6, 128.9, 120.0, 63.8, 54.8, 50.0, 37.3; MS (FAB) m/z (rel. intensity) 468 99), 544 528 472 (13), 471 16), 470 469 468 175 173 88 HRMS (FAB) calcd for C 20 H, C2,N 3 0 4 S+H, 468.0551, found 468.0556.
WO 99/67230 PCT/US99/14233 Example 12.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3ethyl ester (Scheme A, A-8: where RA-I and RA- 2 are the same and equal to H, R 3 is ethyl, Y is CO 2
R
is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is S O H (NSCN '-'OH 0 C o c Example 12 was prepared from example 4 by the procedure described in preparation 6.
Physical properties as follows: mp 118-121 0 C; [a] 2 5 D 106'(c 0.88, ethanol); IR (mull) 3283, 3196, 1665, 1606, 1561, 1539, 1516, 1431, 1414, 1345,1327, 1271, 1219, 1195, 799 cm'; 'H NMR (300 MHz, CD30D) 8 7.59 (2 7.44 (3 7.22 (2 4.69 (1 H), 4.64 (1 4.41 (1 3.24 (3 2.95 (2 1.26 (3 3 C NMR (75 MHz, CDC1 3 8 172.4, 171.5, 163.0, 154.8, 136.7, 136.6, 132.4, 132.1, 130.6, 129.9, 127.9, 120.3, 63.0, 62.7, 53.1, 36.7, 14.3; MS (FAB) m/z (rel. intensity) 540 59), 544 543 (17), 542 540 160 123 118 107 95 23 HRMS (FAB) calcd for C 23
H
23
CL
2
N
3 0 6 S 540.0762, found 540.0730. Anal. Calcd for
C
23
H
3 Cl 2
N
3 0 6 C, 51.12; H, 4.29; N, 7.78. Found: C, 50.77; H, 4.43; N, 7.68.
Example 13.
4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(1,1-dimethylethyl) ester (Scheme A, A-7: where RA-1, and RA-2 are the same and equal to H, R 3 is t-butyl, Y is CO 2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -46- -7 s 0 0 o Example 13 was prepared as described in Scheme A from L-cysteine using di-t-butyl dicarbonate to form the requisite carbamate. Physical data as follows: IR (mull) 1746, 1666, 1606, 1562, 1538, 1516, 1432, 1413, 1324, 1267, 1260, 1216, 1195, 1162, 799 cm'; 'H NMR (CDCl 3 5 7.56 (2 7.46 (1 7.33 (3 7.13 (2 6.94 (1 4.75 (3 H), 4.25 (1 3.75 (3 3.39 (1 3.14 (3 1.43 (9 3 C NMR (DMSO-d 6 8 171.6, 171.5, 170.7, 170.1, 161.7, 152.6, 137.0, 136.9, 136.2, 132.6, 132.6, 131.2, 131.0, 129.3, 128.1, 119.1, 79.7, 78.2, 61.5, 53.4, 53.3, 51.8, 49.3, 49.1, 35.8, 27.6; MS (ESI+) for
C
2 6
H
2 9 Cl 2
N
3 0 6 S m/z 604 MS (ESI-) for C 26
H
2 9 C1 2
N
3 0 6 S m/z 580 Anal.
Calcd for C 26
H
29 C1 2
N
3 0 6 S 0.17 H 2 0: C, 53.34; H, 5.05; N, 7.18. Found: C, 53.47; H, 5.14; N, 7.15. Water 0.51.
Example 14.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3- (1,1-dimethylethyl) ester (Scheme A, A-8: where RA., and RA.
2 are the same and equal to H, R 3 is t-butyl, Y is CO 2
R
5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is N AOH 0 0 C1 c Example 14 was prepared from example 13 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3285, 1665, 1607, 1562, 1538, 1516, 1432, 1413, 1394, 1327, 1259, 1217, 1195, 1162, 799 cm'; 'H NMR (DMSO-d 6 6 12.70 (1 10.67 (1 H), WO 99/67230 WO 9967230PCTIUS99/1 4233 -47- 8.14 (1 7.51 (5 7.20 (2 4.56 (1 4.35 (3 2.98 (3 1.22 (9 3 C NMR (DMS0-l 6 3 172.5, 170.1, 169.9, 161.6, 152.6, 136.9, 136.3, 133.0, 131.2, 131.0, 129.4, 128.1, 119.1, 79.8, 79.7, 61.6, 53.4, 49.2, 48.3, 35.9, 27.6, 20.9; HRMS (FAB) calcd for
C
25
H
27 C1,N 3 0 6 S+H, 568.1075, found 568.1058; MIS (ESI+) for C 25
H
27 C1 2
N
3 0 6 S m/z 567.8 MS (ESI-) for C 25
H
27 C1 2
N
3 0 6 S m/lz 565.8 Anal. Calcd for C 25 H2 7 C1 2
N
3 0 6
S
0.24 H 2 0: C, 52.43; H, 4.84; N, 7.34. Found: C, 52.23; H, 4.76; N, 7.24. Water (KF): 0.75.
Example 5 1 2 ,6-Dichlorobenzoyl)amino]phenyllmethyl]-2-methoxy-2oxoethyl] amino] carbonyl]-3-thiazol idinecarboxylic acid 3-ethyl ester (Scheme A, A-7: where RA-1 and RA-2 are the same and equal to H, R 3 is ethyl, Yis C0 2
R
is 4-[(2,6-dichlorobenzoyl)aminolphenyl and stereochemistry is s H 0 Example 15 was prepared as described in Scheme A from L-cysteine using ethyl chioroformate to form the requisite carbamate. Physical data as follows: IR (mull) 1744, 1666, 1606, 1561, 1538, 1515, 1445, 1431, 1414, 1345, 1325, 1270, 1216, 1194, 1184 cm- I; 'H NMR (CDC1 3 8 7.55 (2 7.36 (4 7.13 (2 6.95 (1 4.74 (3 4.21 (3 3.75 (3 3.40 (1 3.13 (3 1.26 (3 1 3 C NMR (CDC 3 5 171.2, 169.4, 162.3, 136.2, 135.8, 132.4, 132.2, 131.0, 130.3, 130.1, 129.9,128.2, 128.0, 127.9, 120.2, 62.9, 62.7, 53.2, 52.5, 37.1, 14.5, 14.3; MIS (ESI+) for C 24
H
25 C1 2
N
3 0 6 S m/z 553.8 MIS (ESI-) for C 2
,H
25 C1 2
N
3 0 6 S m/z 551.8 Anal. Calcd for C 24
H-
2
C
2
N
3 0 6 S 0.24 H 2 0: C, 51.59; H, 4.60; N, 7.52. Found: C, 51.89; H, 4.62; N, 7.5 1. Water 0.77.
WO 99/67230 PCT/US99/14233 -48- Example 16.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3ethyl ester (Scheme A, A-8: where RA-, and RA-2 are the same and equal to H, R 3 is ethyl, Y is CO 2
R
is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is S 0, ^0 '0 0C S \HO C1 Example 16 was prepared from example 15 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3287, 1664, 1606, 1561, 1539, 1516, 1445, 1431, 1414, 1346, 1327, 1271, 1217, 1195, 799 'H NMR (DMSO-d) 8 12.80 (1 10.69 (1 8.26 (1 7.51 (5 7.19 (2 4.62 (2 4.37 (2 3.94 (2 2.96 (3 H), 1.11 (3 NMR (DMSO-d 6 8 172.5, 169.6, 161.7, 153.5, 136.9, 136.3, 133.0, 131.2, 131.0, 129.5, 128.1, 119.1, 61.2, 53.4, 52.5, 35.9, 22.3, 14.3; MS (ESI+) for
C
23
H
23
C
2
N
3 0 6 S m/z 540.0 MS (ESI-) for C, 2 3
H
23 C1 2
N
3 0 6 S m/z 538.0 HRMS (FAB) calcd for C 23
H,
23 Cl2N 3 0 6 S+H, 540.0762, found 540.0775; Anal. Calcd for
C
2 3
H
23 C1 2
N
3 0 6 S 0.34 HO 2 0: C, 50.54; H, 4.37; N, 7.69. Found: C, 50.53; H, 4.48; N, 7.59.
Water(KF): 1.13.
Example 17.
4 2 ,6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-5,5-dimethyl-3-thiazolidinecarboxylic acid 3-(1,1dimethylethyl) ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to CH 3
R
3 is t-butyl, Y is
CO
2 R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 WO 9967230PCTIUS99/1 4233 -49- N 0
C
0 o Example 17 was prepared as described in Scheme A from L-penicillamine using di-t-butyl dicarbonate to form the requisite carbamnate. Physical data as follows: IR (mull) 1747, 1666, 1606, 1562, 1537, 1516, 1432, 1413, 1324, 1268, 1259, 1213, 1195, 1161, 1142 cm- I NMR (CDC 3 8 7.56 (2 7.34 (4 7.21 (2 6.44 (1 4.94 (1 4.60 (2 H), 4.08 (1 3.70 (3 3.10 (2 1.53 (3 1.42 (9 1.25 (3 NMR (CDC 3 3 171.4, 162.3, 136.3, 135.8, 132.6, 132.4, 131.0, 130.2, 130.0, 128.2, 120.5, 120.3, 120.2, 72.7, 63.9, 60.4, 52.9, 52.3, 48.3, 38.0, 30.3, 28.1, 23.9, 21.0, 14.2; MS (ESI+) for
C
28
H
33 Cl 2
N
3 0 6 S m/z 630.7 Anal. Calcd for C 28
H
33 Cl 2
N
3 0 6 S 0. 13 H 2 0: C, 54.87; H, 5.47; N, 6.86. Found: C, 54.54; H, 5.55; N, 6.54. Water 0.38.
Example 18.
2 6 -Dichlorobenzoyl)aminopheny]methyl]2methoxy-2.
oxoethyl]amino]carbonyl]-3-.thiazolidinecarboxylic acid 3-[(9H-fluoren-1 -yl)methyl] ester (Scheme A, A-7: where RA-1 and RA.
2 are the same and equal to H, R 3 is 9-fluorenylmethyl, Y is C0 2
R
5 is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemnistry is 00 N IkoN Example 18 was prepared as described in Scheme A from D-cysteine using 9fluorenylmethyl chioroformate to form the requisite carbamnate. Physical data as follows: IR (mull) 3280, 1750, 1692, 1671, 1604, 1560, 1538, 1515, 1441, 1430, 1422, 1346, 1320, 1222, 1118 cm 'H NMR (DMSO-1 6 8 8.59 (1 7.87 (2 7.49 (12 4.65 WO 99/67230 PCT/US99/14233 (3 4.26 (4 3.52 (3 2.96 (3 3 C NMR (DMSO-d 6 8 171.4, 161.8, 143.5, 140.6, 137.0, 136.3, 132.5, 131.2, 131.1, 129.5, 128.1, 127.6, 127.1, 125.2, 125.1, 120.0, 119.2, 70.6, 70.0, 63.8, 63.2, 53.3, 53.1, 46.4, 36.2, 25.4; MS (ESI+) for C 36
H
3 1
,C
2
N
3 0 6
S
m/z 703.9 Anal. Calcd for C 36
H
3 ,C1 2
N
3 0 6 S 0.1 H,0: C, 61.23; H, 4.45; N, 5.95.
Found: C, 61.18; H, 4.56; N, 5.89. Water 0.22.
Example 19.
-Carboxy-2-[4-[(2,6dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3- [(9H-fluoren-1-yl)methyl] ester (Scheme A, A-8: where RA-1 and RA-2 are the same and equal to H, R 3 is 9-fluorenylmethyl, Y is CO 2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is S)).PNU- S- O N OH H jr Example 19 was prepared from example 18 by the procedure described in preparation 6.
Physical data as follows: IR(mull) 1672, 1606, 1561, 1533, 1517, 1431, 1413, 1347, 1324, 1269,1218, 1195, 1116, 7 6 0,742 cm NMR (DMSO-d 6 8 10.71 (1 8.32 (1 7.87 (2 7.47 (13 7.16 (2 4.62 (2 4.15 (5 2.90 (4 3 C NMR (DMSO-d 6 8 172.4, 169.2,161.8, 143.6, 140.6,137.0,136.3, 133.0, 131.3, 131.1, 129.7, 129.6. 128.9, 128.2, 127.7, 127.1, 126.8, 125.2,121.3, 120.1, 120.0, 119.2,53.4,48.4, 46.4, 36.4, 29.5, 20.0; MS (ESI+) for C 35
H
2 9 C1 2 N30OS m/z 690.1 Anal. Calcd for
C
3
,H
2 9
C
2
N
3 0 6 S 0.4 H,O: C, 60.25; H, 4.30; N, 6.02. Found: C, 59.88; H, 4.47; N, 5.75.
Water 1.02.
Example 4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-phenylmethyl ester WO 99/67230 PCT/US99/14233 -51- (Scheme A A-7: where RA.I and RA 2 are the same and equal to H, R is phenylmethyl, Y is
CO
2 R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is S- 0 N N
H
o o Example 20 was prepared as described in Scheme A from D-cysteine using benzyl chloroformate to form the requisite carbamate. Physical data as follows: IR (mull) 1748, 1694, 1690, 1673, 1610, 1561, 1542, 1517, 1441, 1430, 1408, 1355, 1324, 1269, 1217 cm 'H NMR (CDC 3 6 7.50 (2 7.33 (9 7.09 (2 6.75 (1 5.19 (2 4.78 (3 4.38 (3 3.73 (3 3.20 (3 3 C NMR (DMSO-d 6 6 171.4, 162.3, 136.3, 135.9, 135.7, 132.5, 132.4, 131.0, 130.0, 128.7, 128.4, 128.2, 128.1, 120.4, 68.2, 63.3, 53.2, 52.5, 37.2; MS (ESI+) for C 29
H
2 7 Cl 2
N
3 0 6 S m/z 637.8 MS (ESI-) for
C
2 9
H
27
CN
3 0 6 S m/z 613.8 Anal. Calcd for C 2 9
H
2 7
,C
2
N
3 0 6 S 0.1 H 2 0: C, 56.39; H, 4.43; N, 6.80. Found: C, 56.31; H, 4.67; N, 6.71. Water 0.19.
Example 21.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3phenylmethyl ester (Scheme A, A-8: where RA-I and RA- 2 are the same and equal to H, R 3 isphenylmethyl, Y is
CO
2 R, is 4-[(2,6-dichlorobenzoyl)amino phenyl and stereochemistry is 0 (SN NQOH 0 co HHj WO 99/67230 PCT/US99/14233 -52- Example 21 was prepared from example 20 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3290, 3034, 1666, 1606, 1562, 1537, 1516, 1431, 1413, 1351, 1326, 1270, 1215, 1195, 799 cm'; 'H NMR (DMSO-d,) 6 12.85 (1 10.65 (1 7.39 (10 7.18 (2 4.98 (2 4.65 (2 4.55 (1 4.33 (1 3.06 (1 H), 2.83 (2 "C NMR (CD 3 OD) 172.6, 163.7,154.4, 136.8, 136.2, 136.1, 131.9, 130.9, 129.6, 128.1, 127.9, 127.8, 127.7, 127.6, 120.0, 67.5, 66.7, 53.4, 36.5; MS (ESI-) for
C
2 8
H
25 C1 2
N
3 0 6 S m/z 599.7 MS (FAB) m/z (rel. intensity) 602 99), 678 604 603 602 560 558 468 466 371 91 HRMS (FAB) calcd for C 28
H
25 C1 2
N
3 0 6 S+H, 602.0919, found 602.0913; Anal. Calcd for C 28
H
25 C12N 3 0 6 S 0.23 H 2 0: C, 55.45; H, 4.23; N, 6.93. Found: C, 55.53; H, 4.46; N, 6.88. Water 0.67.
Example 22.
2 ,6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(tricyclo[3.3.1.13, 7 ]dec-l-yl) ester (Scheme A, A-7: where RA., and RA 2 are the same and equal to H, R 3 is 1-adamantyl, Y is CO2, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is so o c N N Example 22 was prepared as described in Scheme A from D-cysteine using 1-adamantyl fluoroformate to form the requisite carbamate. Physical data as follows: IR (mull) 3284, 3271, 1747, 1690, 1684, 1666, 1557, 1532, 1436, 1412, 1355, 1298, 1194, 1053, 799 cm-'; 'H NMR (DMSO-d 6 8 10.67 (1 8.41 (1 7.53 (5 7.17 (2 4.51 (3 4.23 (1 3.63 (3 3.22 (1 3.04 (1 2.90 (1 2.75 (1 2.01 (9 1.56 (6 H); "C NMR (DMSO-d) 8 172.1,162.3, 152.7, 137.6, 136.9, 133.2, 131.8, 131.6, 130.0, 128.7, 119.8, 53.8, 52.4, 36.7, 36.1, 30.6; MS (ESI+) for C 3 2
H
3 ,C1 2
N
3 0 6 S m/z 659.7 WO 99/67230 PCT/US99/14233 -53- MS (ESI-) for C 3 2
H
3 ,Cl2N 3 0 6 S m/z 657.7 MS (FAB) m/z (rel. intensity) 660 662 660 618 616 480 173 136 135 123 93 Anal. Calcd for C 3 2
H
3 ,C1,N 3 0 6 S 0.04 HO 2 0: C, 58.12; H, 5.35; N, 6.35.
Found: C, 58.19; H, 5.62; N, 6.25. Water 0.10.
Example 23.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3- (tricyclo[3.3.1.1 3, 7 ]dec-1-yl) ester (Scheme A, A-8: where RA., and RA- 2 are the same and equal to H, R 3 is 1-adamantyl, Y is
CO
2 R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is 2 o- oc N 0O 0- O 0 0 YZ cl Example 23 was prepared from example 22 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3287, 1667, 1606, 1562, 1537, 1516, 1431, 1412, 1353, 1326, 1299, 1274, 1220, 1194, 1049 'H NMR (DMSO-d 6 6 12.70 (1 H), 10.67 (1 8.21 (1 7.58 (5 7.17 (2 4.51 (3 4.22 (1 3.20 (1 3.04 (1 2.88 (1 2.76 (1 2.07 (9 1.12 (6 3 C NMR (DMSO-d 6 8 173.1, 162.3, 152.7, 137.5, 136.9, 133.8, 131.8, 131.6, 130.1, 128.7, 119.7, 80.0, 53.8, 36.8, 36.1, 30.6; MS (ESI+) for C 3
,H
33 Cl 2
N
3 0 6 S m/z 645.8 HRMS (FAB) calcd for
C
3
,H
33 Cl2N 3 0 6 S+H, 646.1545, found 646.1564; Anal. Calcd for C 3
,H
33 Cl2N 3 0 6 S 0.29
H
2 0O: C, 57.13; H, 5.19; N, 6.45. Found: C, 56.82; H, 5.21; N, 6.32. Water 0.80.
Example 24.
4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[2-(4-morpholinyl)ethyl] ester WO 99/67230 WO 9967230PCTIUS99/1 4233 -54- (Scheme A, A-7: where RA.I and RA.
2 are the same and equal to H, R 3 is 2-(4morpholinyl)ethyl, YS is G 2
R
5 is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemnistry is s- 0 o Example 24 was prepared as described in Scheme A from D-cysteine using 4-(2hydroxyethyl)morpholine to form the requisite carbamnate. Physical data as follows: IR (mull) 1745, 1705, 1679, 1605, 1536, 1515, 1431, 1414, 1344, 1323, 1269, 1216, 1194, 1183, 1117 cm 1 1 H NMR (300 MHz, CDCI 3 8 7.82 (1 7.56 (2 7.31 (3 7.12 (2 6.95 (1 4.65 (3 4.39 (3 3.74 (3 3.69(3 3.34 (1 3.18 (3 H), 2.763 (5 3 C NMR (75 MHz, CDC1 3 6 171.5, 169.8, 162.9, 154.4, 136.8, 136.1, 132.1, 130.6, 129.7, 127.9, 120.2, 66.3, 62.7, 57.0, 53.4, 53.2, 52.4, 36.7, 29.5; MIS (ESI-) for C 28
H
32 C1 2
N
4 0 7 S m/z 636.8 HRMS (FAB) calcd for C 28
H
32
CL
2
N
4 0 7 S +H, 639.1447, found 639.1419. Anal. Calcd for C 28
H
32
C,N
4 0 1 5: C, 52.58; H, 5.04; N, 8.76.
Found: C. 52.47; H, 5.17; N, 8.69.
Example dichlorobezoyI)amino~phenyl~ethyl~amino]carbonyl]-3thiaolidinecarboxylic acid 3- 2 -(4-morpholinyl)ethyl] ester (Scheme A, A-8: where RA-1 and RA- 2 are the same and equal to H, R 3 is 2-(4morpholinyl)ethyl, Y is CO 2
R
5 is 4 2 ,6-dichlorobenzoyl)aminolphenyI and stereochemnistry is WO 99/67230 PCT/US99/14233 S- 0 00 0
H-
Example 25 was prepared from example 24 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3278, 1667, 1606, 1562, 1541, 1515, 1431, 1413, 1351, 1326, 1270, 1195, 1134, 1118, 799 'H NMR (300 MHz, DMSO-d 6 5 10.64 (1 8.25 (1 7.50 (5 7.16 (2 4.60 (2 4.44 (1 4.27 (1 4.06 (2 H), 3.51 (4 3.43 (2 3.29 (4 2.42 (4 3 C NMR (75 MHz, DMSO-d 6 8 173.0, 172.4, 169.8, 162.3, 137.5, 136.8, 133.5, 131.7, 130.1, 128.6, 119.7, 66.6, 63.3, 57.0, 53.7, 36.7, 21.5; MS (FAB) m/z (rel. intensity) 625 55), 629 628 627 (39), 626 625 308 141 114 113 100 HRMS (FAB) calcd for
C
2 7
H
30
CLN
4 0,S 625.1290, found 625.1309.
Example 26.
4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(1,1-dimethylethyl) ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to H, R 3 is t-butyl, Y is CO 2
R
5 is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is S-
H
NYN
O N Cl Example 26 was prepared as described in Scheme A from D-cysteine using di-t-butyl dicarbonate to form the requisite carbamate. Physical data as follows: IR (mull) 3293, 1746, 1666, 1606, 1562, 1538, 1516, 1432, 1413, 1324, 1260, 1216, 1195, 1162, 799 cm'; 'H NMR (CDCI 3 8 7.55 (2 7.37 (4 7.14 (2 4.89 (1 4.66 (2 4.25 (1 H), 3.75 (3 3.39 (1 3.24 (3 1.44 (9 3 C NMR (CDCl 3 8 171.2, 169.8, 162.3, WO 99/67230 PCT/US99/14233 -56- 136.3, 135.8, 132.4, 132.3, 131.0, 130.2, 130.1, 129.9, 128.2, 128.1, 127.9, 120.5, 120.2, 120.1, 82.2, 53.2, 52.5, 37.3, 31.0, 28.4, 28.2; MS (ESI+) for C 26
H
29
C
2
N
3 0 6 S m/z 603.9 MS (ESI-) for C 26
H
2 9
CI
2
N
3 0 6 S m/z 580.0 HRMS (FAB) calcd for
C
26
H
29 ,Cl2N 3 0 6 S+H, 582.1232, found 582.1231. Anal. Calcd for C 26
H
29 C1 2
N
3 0 6 S 0.26
H
2 0: C, 53.18; H, 5.07; N, 7.16. Found: C, 52.78; H, 5.14; N, 6.91. Water 0.66.
Example 27.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3- (1,1-dimethylethyl) ester (Scheme A, A-8: where RA, and RA-2 are the same and equal to H, R 3 is t-butyl, Y is CO 2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is H O
H
N
H~
0oo o a Example 27 was prepared from example 26 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3286, 1665, 1606, 1562, 1538, 1516, 1432, 1413, 1394, 1326, 1259, 1216, 1195, 1161, 799 cm'; 'H NMR (DMSO-d 6 8 12.70 (1 10.63 (1 8.13 (1 7.51 (5 7.20 (2 4.45 (4 2.96 (3 1.23 (9 3 C NMR (DMSO-d 6 173.1, 170.5, 162.3, 153.2, 137.5, 136.9, 131.8, 131.6, 130.0, 128.7, 119.7, 80.4, 62.2, 54.0, 49.8, 36.5, 28.3, 21.5; MS (ESI+) for C 25
H
2 7
C
2
N
3 0 6 S m/z 567.9 MS (ESI-) for C 25
H
27
C
2
N,
3 0 6 S m/z 565.9 HRMS (EI) calcd for C 25
H
27 C1 2
N
3 0 6
S
567.0997, found 568.1096. Anal. Calcd for C 25
H
27 C1 2
N
3 0 6 S 0.34 H 2 0: C, 52.82; H, 4.79; N, 7.39. Found: C, 52.17; H, 4.90; N, 7.25. Water 1.07.
Example 28.
2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-5,5-dimethyl-3-thiazolidinecarboxylic acid 3-(1,1dimethylethyl) ester WO 99/67230 PCT/US99/14233 -57- (Scheme A, A-7: where RA-1 and RA- 2 are the same and equal to CH 3
R
3 is t-butyl, Y is
CO
2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is H 0 Example 28 was prepared as described in Scheme A from D-penicillamine using di-t-butyl dicarbonate to form the requisite carbamate. Physical data as follows: IR (mull) 1744, 1707, 1688, 1678, 1657, 1606, 1562, 1541, 1516, 1431, 1414, 1326, 1253, 1161, 1140cm 'H NMR (CDC 3 6 7.58 (2 7.36 (3 7.14 (2 6.55 (1 4.87 (1 4.56 (2 H), 4.10 (1 3.72 (3 3.08 (2 1.53 (3 1.44 (9 1.40 (3 3 C NMR (CDC 3 8 171.6, 162.3, 153.6, 136.4, 135.8, 132.6, 132.4, 131.0, 130.1, 129.9, 129.8, 128.2, 120.7, 120.5, 120.3, 81.7, 73.0, 52.8, 52.6, 52.4,48.4, 39.8, 39.5, 37.6, 30.3, 28.3, 28.0, 23.5; MS (ESI-) for C 28
H
33
C
2
N
3 0 6 S m/z 607.9 MS (FAB) m/z (rel. intensity) 610 512 510 117(30), 115 99 87(16), 59 57 57 41 HRMS (FAB) calcd for C 28
H
33 Cl2N 3 0 6 S+H, 610.1545, found 610.1501; Anal. Calcd for C 2
,H
33 C1 2
N
3 0 6 ,S 0.07 H 2 0: C, 54.97; H, 5.46; N, 6.87. Found: C, 54.92; H, 5.54; N, 7.11. Water 0.21.
Example 29.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-5,5-dimethyl-3thiazolidinecarboxylic acid 1-dimethylethyl) ester (Scheme A, A-8: where RA-1, and RA-2 are the same and equal to CH 3
R
3 is t-butyl, Y is
CO
2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -58- N ^OH s0-0 o o Example 29 was prepared from example 28 by the procedure described in preparation 6.
Physical data as follows: IR(mull) 1739, 1666, 1606, 1562, 1535, 1516, 1432, 1413, 1394, 1325, 1270, 1260, 1194, 1160, 799 'H NMR (DMSO-d 6 8 12.62 (1 10.62 (1 8.18 (1 7.51 (5 7.19 (2 4.50 (3 4.24 (1 2.91 (2 1.33 (12 H), 1.04 (3 3 C NMR (DMSO-d 6 8 172.8, 168.7, 161.9, 153.2, 137.2, 136.5, 133.1, 131.4, 131.3, 129.6, 128.3, 119.5, 80.0, 70.9, 53.7, 48.5, 37.0, 30.7, 28.1, 27.9, 24.6; MS (ESI+) for C 27
H
31
CI
2
N
3 0 6 S m/z 595.9 MS (ESI-) for C 27
H
31 C1 2
N
3 OS m/z 593.8 MS (FAB) m/z (rel. intensity) 596 19), 672 596 499 498 60), 497 496 494 173 116 57 Anal. Calcd for C 27
H
31
CI
2
N
3 0 6
S
0.27 H 2 0: C, 53.93; H, 5.29; N, 6.99. Found: C, 53.73; H, 5.39; N, 7.10. Water (KF): 0.80.
Example 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-5,5-dimethyl-3-thiazolidinecarboxylic acid 3-ethyl ester (Scheme A, A-7: where RA.- and RA 2 are the same and equal to CH 3
R
3 is ethyl, Y is CO 2
R
5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is s+ H 0 N 0 c Example 30 was prepared as described in Scheme A from D-penicillamine using ethyl chloroformate to form the requisite carbamate. Physical data as follows: IR (mull) 3292, 1748, 1666, 1606, 1562, 1538, 1516, 1445, 1431, 1414, 1341, 1325, 1271, 1212, 1194 cm WO 99/67230 WO 9967230PCTIUS99/1 4233 -59- NMIR (CDC1 3 6 7.57 (2 7.37 (4 7.11 (2 6.49 (1 4.86 (1 4.59 (2 H), 4.14 (3 3.75 (3 3.09 (2 1.60 (3 1.54 (3 1.23(3 3 C NMR (CDC1 3 6 171.6, 162.3, 154.5, 136.3, 135.8, 132.6, 132.4, 131.1, 130.2, 130.1, 129.8, 128.2, 120.5, 120.4, 72.7, 62.5, 52.7, 52.5, 37.3, 30.2, 23.7, 14.6, 14.1; MS (ESI+) for C 26
H
29 C1 2
N
3 0 6 m/z 581.9 MS (ESI-) for C 26
H
29 C1 2
N
3 0 6 S m/z 579.8 HRMS (El) calcd for
C
26
H
29 C1 2
N
3 0 6 S 581.1154, found 581.1132; Anal. Calcd for C 26
H
29 C1 2
N
3 0 6 S 0. 16 H 2 0: C, 53.35; H, 5.05; N, 7.18. Found: C, 53.74; H, 5.12; N, 7.12. Water 0.49.
Example 3 1.
dichlorobenzoyl)amino]pheny]ethyI amino] carbonyl],-imethy-3thiazolidinecarboxylic acid 3-ethyl ester (Scheme A, A-8: where RA., and RA.
2 are the same and equal to CH3 R 3 ietyYis C0 2 1 R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemistry is 0 C C1 Example 31 was prepared from example 30 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3287, 3070, 1666, 1606, 1562, 1538, 1516, 143 1, 1414,1342,1328,1271,1213,l194, 799cm-1; 'HNMR(DMSO-d 6 )5 12.58 (1 10.63 (1 8.22 (1 7.51 (5 7.20 (2 4.51 (3 3.95 (3 3.04 (1 2.86 (1 H), 1.3 5(3 1. 16 1 3 CNMR (DMSO-d 6 )8 223.3, 184.1, 183.9, 172.8, 168.4, 161.7, 136.9, 136.3, 133.1, 131.2, 131.0, 129.4, 128.1, 119.2, 70.6, 61.1, 53.6, 53.4, 48.6, 36.2, 30.1, 25.4, 24.3, 21.0, 14. 1; MS (ESI+) for C 2 5
H
27 C1 2
N
3 0 6 5 m/z 568.0 MIS (ESI-) for C 25
H
27 C1 2
N
3 0 6 S m/z 565.9 MS (FAB) m/z (rel. intensity) 568 86), 644 571 570 569 568 335 188 173 141 116 HRMS (FAB) calcd for C 25 H2 7 C1 2
N
3 0 6 S+H, 568.1075, found 568.1096; Anal. Calcd WO 99/67230 PCT/US99/14233 for C 25
H
27 jC 2
N
3 0 6 S 0.4 H 2 0: C, 52.16; H, 4.87; N, 7.30. Found: C, 52.46; H, 4.90; N, 7.15. Water 1.25.
Example 32.
2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(1,1-dimethylethyl) ester (Scheme A, A-7: where RA.I and RA-2 are the same and equal to H, R 3 is t-butyl, Y is CO 2 R, is 4 -[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is 0 0- c N O O OO CI Example 32 was prepared as described in Scheme A from D-cysteine using di-t-butyl dicarbonate to form the requisite carbamate. Physical data as follows: IR (liq.) 1745, 1702, 1565, 1511, 1467, 1439, 1368, 1299, 1241, 1197, 1177, 1162, 1017, 778, 768 cm'; 'H NMR (CDCl 3 8 7.36 (2 7.25 (1 7.06 (2 6.94 (2 5.23 (2 4.75 (3 H), 4.12 (1 3.72 (3 3.37 (1 3.14 (3 1.45 (9 13 C NMR (CDC1 3 8 171.5, 157.9, 136.9, 132.0, 130.3, 128.4, 128.2, 115.0, 81.9, 65.1, 62.8, 53.2, 52.3, 49.2, 36.9, 28.1, 27.9; MS (ESI+) for C 26
H
3 0 ClN 2 0 6 S m/z 568.9 MS (ESI-) for
C
26
H
3 0 C1 2
N
2 0 6 S m/z 566.7 Anal. Calcd for C 26
H
3 C1 2
N
2 0 6 OS 0.09 H 2 0: C, 54.68; H, 5.33; N, 4.91. Found: C, 54.62; H, 5.41; N, 4.73. Water 0.28.
Example 33.
dichlorophenyl)methoxy]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3- (1,1 -dimethylethyl) ester (Scheme A, A-8: where RA-1 and RA.
2 are the same and equal to H, R 3 is t-butyl, Y is CO 2 R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -61o N N OH Example 33 was prepared from example 32 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1734, 1704, 1676, 1612, 1565, 1511, 1439, 1393, 1300, 1241, 1196, 1178, 1162, 777, 769 cm'; 'H NMR (DMSO-d 6 8 8.24 (1 7.50 (3 7.13 (2 6.93 (2 5.16 (2 4.52 (3 4.21 (1 2.91 (4 1.31 (9 H); 13C NMR (DMSO-d 6 172.7, 169.7, 157.1, 152.6, 135.9, 131.7, 131.5, 130.2, 129.8, 128.7, 114.2, 79.8, 64.8, 61.6, 53.2, 49.2, 36.0, 34.8, 27.8, 21.0; MS (ESI+) for
C
25
H
28 C1 2
N
2 0 6 S m/z 554.9 MS (ESI-) for C 2 5
H
28 C1 2
N
2 0 6 S m/z 552.8 Anal. Calcd for C, 5
H
28
CI
2
N
2 0 6 S 0.15 H 2 0: C, 53.79; H, 5.11; N, 5.02. Found: C, 54.17; H, 5.17; N, 5.00. Water 0.50.
Example 34.
2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to H, R 3 is ethyl, Y is CO2-, Rs is 4 2 6 -dichlorophenyl)methoxy]phenyl and stereochemistry is o o 0 0 ci Example was prepared as described in Scheme A from D-cysteine using ethyl chloroformate to form the requisite carbamate. Physical data as follows: IR (mull) 3282, 1742, 1705, 1692, 1664, 1562, 1509, 1436, 1352, 1343, 1236, 1196, 1175, 1015, 786 cm-; 'H NMR (CDCl 3 5 7.37 (2 7.25 (1 7.05 (2 6.94 (2 6.74 (1 5.23 (2 H), WO 99/67230 PCT/US99/14233 -62- 4.77 (3 4.34 (1 4.18 (2 3.74 (3 3.37(1 3.13 (3 1.31 (3 1 3 C NMR
(CDCI
3 5 171.6, 171.4, 158.0, 137.0, 132.1, 130.5, 130.4, 128.5, 128.2, 115.0, 65.2, 63.6, 63.2, 62.7, 53.6, 53.2, 52.4, 36.9, 14.5; MS (ESI+) for C 24
H
2 6 C1 2
N
2 0 6 S m/z 540.9 HRMS (EI) calcd for C 24
H
2 6
CI,
2
N
2 0 6 S 540.0889, found 540.0878; Anal. Calcd for
C
24
H
2 6 C1 2
N
2 0 6 S 0.26 H 2 0: C, 52.79; H, 4.89; N, 5.13. Found: C, 52.41; H, 4.82; N, 4.96.
Water 0.85.
Example -carboxy-2-[4-[(2,6dichlorophenyl)methoxy]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3ethyl ester (Scheme A, A-8: where RA,- and RA-2 are the same and equal to H, R 3 is ethyl, Y is C0 2 R, is 4 2 ,6-dichlorophenyl)methoxy]phenyl and stereochemistry is N OH oo Example 35 was prepared from example 34 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1709, 1675, 1612, 1565, 1511, 1439, 1416, 1346, 1300, 1241, 1196, 1179, 1115, 1018, 768 'H NMR (CDCl 3 8 7.34 (2 7.23 (1 H), 7.12 (2 6.94 (2 6.85 (1 5.22 (2 4.77 (4 4.34 (1 4.16 (2 3.33 (4 1.26 (3 3 C NMR (CDC1 3 8 174.0, 170.2, 158.1, 155.1, 137.0, 132.0, 130.5, 128.5, 128.1, 115.0, 65.2, 63.9, 63.0, 62.9, 53.3, 36.4, 21.9, 14.5; MS (ESI+) for
C
23
H
24
C
2
N
2 0 6 S nm/z 527.0 MS (ESI-) for C,2 3
H
24
C
2
N
2 0 6 S m/z 524.9 HRMS (EI) calcd for C 23
H
24
C
2
N
2 0 6 S 526.0732, found 526.0726; Anal. Calcd for WO 99/67230 PCT/US99/14233 -63-
C
23
H
24 C1 2
N
2 0 6 S 0.20 H 2 0: C, 52.02; H, 4.63; N, 5.27. Found: C, 52.12; H, 4.73; N, 5.34.
Water 0.69.
Example 36.
1 4 -[(2,6-Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3 2 4 -morpholinyl)ethyl] ester (Scheme A, A-7: where RA-I and RA- 2 are the same and equal to H, R 3 is 2-(4morpholinyl)ethyl, Y is C0 2 R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is Example 36 was prepared as described in Scheme A from D-cysteine using 4-(2hydroxyethyl)morpholine to form the requisite carbamate. Physical data as follows: mp 138-140 0 C; JR (mull) 3286, 1743, 1705, 1660, 1559, 1513, 1435, 1428, 1302, 1245, 1226, 1215, 1176, 1015, 764 cm 'H NMR (300 MHz, DM0-l 6 8 8.43 (1 7.48 (3 H), 7.13 (2 6.94 (2 5.16 (2 4.59 (2 4.48 (1 4.26 (1 4.07 (2 3.63 (3 3.51 (4 3.23 (1 3.01 (1 2.84 (1 2.71 (1 2.41 (6 'C NMR MHz, DMSO-d) 8 172.2, 157.7, 153.9, 136.5, 132.2, 132.0, 130.8, 130.1, 129.2, 114.8, 66.6, 65.4, 63.4, 57.0, 53.9, 53.8, 52.4, 36.4; MS (ESI+) for C 28
H
33 C1N 3 0 7 5 in/z 625.8 Anal. Calcd for C 28
H
33 C1 2
N
3 0 7 5: C, 53.67; H, 5.31; N, 6.71. Found: C, 53.69; H, s-5.27; N, 6.69.
O N 'O' 37.6 was prepared as described in Scheme A from D-cysteine using 4-(2- I -C arboxy-2-[4-[(2,6dichloydroxyhenyl)mrehoxyline to form nylthyl]ae requisite carbaonyl]3ate. Physiolidinecarboxylic alows:cid 3-mp 15 138-140 2 IR (mull) 3286, 1743, 1705,-morphoinyl)ethy]1660, 1559, 1513, 1435,]1428, 1302, 1245, 1226,ester 1215, 1176, 1015, 764 cm I1H NMR (300 MHz, DMSO-d6) 8 8.43 (1 7.48 (3 H), 7.13 (2 6.94 (2 5.16 (2 4.59 (2 4.48 (1 4.26 (1 4.07 (2 3.63 (3 3.51 (4 3.23 (1 3.01 (1 2.84 (1 2.71 (1 2.41 (6 "C NMR MHz, DMSO-d6) 8 172.2, 157.7, 153.9, 136.5, 132.2, 132.0, 130.8, 130.1, 129.2, 114.8, 66.6, 65.4, 63.4, 57.0, 53.9, 53.8, 52.4, 36.4; MS (ESI+) for C28H33Cl2N307S m/z 625.8 Anal. Caled for C28H33Cl2N307S: C, 53.67; H, 5.3 1; N, 6.71. Found: C, 53.69; H, 5.27; N, 6.69.
Example 37.
dichlorophenyl)methoxy]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3- [2-(4-morpholinyl)ethyl] ester WO 99/67230 PCT/US99/14233 -64- (Scheme A, A-8: where RA.I and RA.2 are the same and equal to H, R 3 is 2-(4morpholinyl)ethyl, Y is CO 2 R, is 4 2 6 -dichlorophenyl)methoxy]phenyl and stereochemistry is o O N N OH 0 C 00 o ooi a o Example 37 was prepared from example 36 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1710, 1610, 1585, 1565, 1511, 1439, 1408, 1351, 1301, 1240, 1196, 1179, 1116, 1017, 767 cm 1 H NMR (300 MHz, DMSO-d 6 8 8.27 (1 7.55 (2 7.45 (1 7.13 (2 6.94 (2 5.16 (2 4.59 (2 4.40 (1 H), 4.27 (1 4.04 (2 3.52 (4 3.21 (1 2.86 (3 2.44 (6 "C NMR (75 MHz, DMSO-d 6 5 173.2, 172.5, 157.6, 136.5, 132.2, 132.0, 130.8, 130.4, 129.2, 114.7, 66.5, 65.3, 63.2, 57.0, 53.9, 53.7, 36.5, 21.5; MS (ESI+) for C 2 7
,H
3 ,C1 2
N
3 0,S m/z 611.9 Anal. Calcd for C, 27
H
3 ,C1 2
N
3 0 7 S 1.0 C 2
H
4 0 2 0.63 H 2 0 0.28 HCI: C, 50.13; H, 5.31; N, 6.03; Cl, 11.59. Found: C, 49.80; H, 5.30; N, 6.05; Cl, 11.20. Water 1.58.
Example 38.
1-[ 4 2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[(2-pyridinyl)methyl] ester (Scheme A, A-7: where RA., and RA- 2 are the same and equal to H, R3 is 2-pyridinylmethyl, Y is CO 2 R, is 4 2 6 -dichlorophenyl)methoxy]phenyl and stereochemistry is H0
O
o oo WO 99/67230 PCT/US99/14233 Example 38 was prepared as described in Scheme A from D-cysteine using 2pyridinemethanol to form the requisite carbamate. Physical data as follows: mp 123- 125 0 C; IR (mull) 3334, 1728, 1709, 1668, 1531, 1511, 1441, 1405. 1345. 1294, 1286, 1236, 1228, 1015, 762 cm 'H NMR (300 MHz, DMSO-d 6 8 8.57 (2 7.79 (1 H), 7.54 (2 7.42 (1 7.27 (2 7.12 (2 6.92 (2 5.13 (4 4.69 (2 4.49 (1 4.34 (1 3.59 (3 3.24 (1 2.89 (3 'C NMR (75 MHz. DMSO-d 6 172.2. 169.9, 157.7, 156.5, 153.6, 137.3, 136.5, 132.2, 132.0, 130.8. 130.1, 129.2. 123.2, 121.0, 114.8, 67.7, 65.3, 62.0, 54.0. 52.4, 50.3, 36.3, 35.3; HRMS (El) calcd for
C,,H,
7 ClN 3 0 6 S 603.0997. found 603.0992; Anal. Calcd for C 2 8 7
CN
3 0 6 S: C, 55.63; H, 4.50; N. 6.95. Found: C. 55.56: H, 4.59; N, 6.93.
Example 39.
[S-(R*,R*)]-4-[[[1-[4-[(2,6-Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[2-(1-pyrrolidinyl)ethyl] ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to H. R 3 is 2-(1pyrrolidinyl)ethyl, Y is R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is
O
N N 0 00 ci Example 39 was prepared as described in Scheme A from D-cysteine using 1-(2hydroxyethyl)pyrrolidine to form the requisite carbamate. Physical data as follows: mp 130-132 0 C; IR (mull) 1745. 1702, 1661, 1556, 1513, 1435, 1426, 1303, 1245, 1226, 1214, 1176, 1017, 825, 765 cm 'H NMR (300 MHz, DMSO-d 6 5 8.42 (1 7.48 (3 7.13 (2 6.94 (2 5.16 (2 4.59 (2 4.48 (1 4.27 (1 3.99 (2 3.63 (3 H), 3.24(1 2.68 (9 1.62 (4 3 "C NMR (75 MHz, DMSO-d 6 8 172.2, 170.0, 157.7, 136.5, 132.2. 132.0, 130.8. 130.1. 129.2, 114.8, 65.3, 65.1, 54.4. 53.9. 52.42, 36.35, 23.6; WO 99/67230 WO 9967230PCTIUS99/1 4233 -66- MS (ESI+) for C, 8
H
33 C1 2
N
3 0 6 S m/lz 609.8 Anal. Calcd for C, 8
H
3 3 C1 2
N
3 0 6 S: C, 55.08; H. 5.45; N. 6.88. Found: C. 54.72; H. 5.58; N. 6.60.
Example .S I-[ 4 2 6 -Dichlorophenvl)methoxy]phenylmethyl]2methoxy-2.
oxoethyl]aminolcarbonylI-3-thiazolidinecarboxyI ic acid 3-(l,--dimethylethyl) ester (Scheme A. A-7: where RA., and RA.
2 are the same and equal to H, R, is t-butyl, Y is C0 2 R, is 4 -[(2,6-dichlorophenyl)methoxylphenyl and stereochemnistry is STJ).
sN 0K- Example 40 was prepared as described in Scheme A from L-cysteine using di-t-butyl dicarbonate to form the requisite carbamate. Physical data as follows: IR (mull) 1746, 1702, 1611. 1565, 1511, 1439, 1299, 1241, 1197, 1177, 1162,1118, 1016, 777, 768 cmn"'; 'H NMR (CDCI 3 5 7.39 (2 7.25 (1 6.93 (2 6.93 (2 5.24 (2 4.72 (3 H), 4.20 (1 3.74(3 3.35 (1 3.12(3 1.44 (9 3 C NMR (CDCl 3 8 171.7, 170.0, 169.5. 158.3, 137.3, 132.4, 130.7. 130.6. 128.8, 128.5, 115.2, 115.1, 82.4. 65.5, 53.5. 52.6. 50.3. 37.3, 28.4; MS (ESI+) for C', 6
H
30 C1,N 2 -0 6 S m/z 554.9 MS (ESI-) for C- 6
H
3
CI,NO
6 S m/lz 552.8 Anal. Calcd for C, H 3 CN0S 0. 1 H,O: C, 54.65; H. 5.33; N. 4.90. Found: C, 54.59; H. 5.30, N, 4.88. Water 0.33.
Example 4 1.
,S -Carboxy-2-[4-[(2,6dichlorophenvl)methoxlphenyl]ethyl]amino]carbonyl-3-thiazolidinecarboxylic acid 3- (1 ,1-dimethylethyl) ester (Scheme A. A-8: where RI and RA., are the same and equal to H, R 3 is t-butyl, Y is R, is 4-[(2.6-dichlorophenyl)methoxy]phenvl and stereochemnistry is WO 99/67230PCIS9143 PCTIUS99/14233 -67- 7 K, 0 C' Example 41 was prepared from example 40 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1737, 1705, 1679, 1612, 1565, 1512, 1439, 1300, 1241, 1196, 1178, 1163, 1117, 777, 769 cm-; 'H NMR (DMSO-1 6 )56 8.07 (1 7.54 (2 7.44 (1 7.16 (2 6.93 (2 5.16 (2 4.56 (1 4.39 (3 3.56 (1 H), 2.84 (3 1.23 (9 H); 3 CNMR (DMSO-d.).8 172.7, 169.9, 157.0, 152.6, 135.9, 131.6, 131.4. 130.1, 129.9, 129.8, 128.7, 114.1, 79.8, 66.9. 64.7, 61.7, 53.5, 49.3, 35.6, 34.8, 27.9, 27.6; MS (ESI+) for C 2 5 28
C,N
2 0 6 5 m/z 554.9 (M+H) t MS (ESI-) for C,-1 2 8 C1-,N -0 6 S m/z 552.8 Anal. Calcd for C 25
H.,
8 C1 2 N,0 6 S 0.27 C. 53.59; H, 5.13; N, 5.00. Found: C, 53.97; H, 5.14; N, 4.96. Water 0.86.
Example 42.
4 2 6 -Dichlorobenzoyl)amino]-N-[[(4S)-3-(ethylsulfonyl)-4thiaolidinylcarbony].
L-phenylalanine methyl ester (Scheme A, A-7: where RA.1 and RA- 2 are the same and equal to H. R 3 is ethyl,Y is SO 2
R
is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is
H
c Example 42 was prepared as described in Scheme A from D-cysteine using ethanesulfonyl chloride to form the requisite sulfonamide. Physical data as follows: IR (mull) 1743, 1666, 1605, 1561, 1535. 1515. 1432. 1413, 1328, 1269. 1219. 1195. 1146. 799. 782 cm WO 99/67230 WO 9967230PCT/US99/I 4233 -68- 'H NMR (CDCI 3 )8 7.58 (2 7.46 (1 7.32(3 7.17 (2 7.07 (1 4.85 (1 H), 4.73 (2 4.28 (1 3.76(3 3.54 (1 3.26 (1 3.05 (4 1.40 (3 1 3 C NMR (CDC1 3 6 171.3, 168.2. 162.4, 136.4 135.8. 132.4. 132.3, 131.0, 130.1. 128.2. 120.6, 64.9, 53.2. 52.6, 51.5. 45.8, 37.1, 34.2. 3 1.0. 29.3. 7.7; MS (ESI+) for C, H-, 5
C,N
3 5 tnZ 573.9 MS (ESI-) for C 23
H
25
C,N.
3 0 6
S
2 nm/z 571.7 HRMS (FAB) calcd for C 2 3
H
25 C1,N 3 0 6
S
2 574.0640, found 574.0634; Anal. Calcd for C2 3
H,
5 C1 2
N
3 0 6 5.,'0.1
H
2 0: C, 47.97; H, 4.40; N, 7.30. Found: C. 48.36; H, 4.59; N, 6.80.
Example 43 4-(,-ihooezy~mn]N[[4 34ehlufnl haoidinyicarbonyl]- L-phenylalanine (Scheme A, A-8: where RA,, and RA- 2 are the same and equal to H, R3 is ethyl, Y is SOD, is 4 2 6 -dichlorobenzoyl)amino]phenyI and stereochemnistry is s- H0
OH
o=s=o 0 0 ci Et Example 43 was prepared from example 42 by the procedure described in preparation 6..
Physical data as follows: IR (mull) 17-34, 1664. 1605, 1562, 1536, 1516, 1432, 1414, 1330, 1272.,1234, 1195, 1146, 799, 781 'H NMR (DMSO-d 6 6 12.91 (1 10.65 (1 8.15 (1 7.51 (5 7.17 (2 4.77 (2 4.43 (1 4.29 (1 3.94 (6 H), 1.20 (3 1 3 CNMR (DMSO-d 6 )86 172.5. 168.9, 161.9, 137.1, 136.5. 133.2. 131.4.,131.3, 129.8, 128.3. 119.3, 63.7, 53.6, 51.4, 45.3. 3 6.1, 34.7; MS (ESI+) for C,I1 3 Cl 2
N
3
O
6
S
2 m/z 559.9 MIS (ESI-) for 3 C1 2 N,0 6 S, rn/z 557.8 HRMS (FAB) calcd for
CH.
3 C1,N 3 0 6
S
2 560.0483, found 560.0488; Anal. Calcd for CVHCIN0O 6 S -0.72 H,O: C. 46.08; H. 4.30; N, 7.33. Found: C. 46.42- H, 4.37; N, 7.01. Water 2.26.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -69- Example 44.
4-[(2,6-Dichlorobenzovl)amino]-N-[ [(45 [5-(trifluoromethyl)-2pyridinyl]sulfonyl]-4-thiazolidinyl]carbonv ]-L-phenylalanine methyl ester (Scheme A, A-7: where R, and R,- 2 are the same and equal to H. R 3 is triflouromethylpyridvi). Y is SO,. R, is 4 -[(2,6-dichlorobenzoyI)amino~phenyl and stereochemnistry is S. H N N H
CF
3 Example 44 was prepared as described in Scheme A from D-cysteine using triflouromethvlpyridyl)sulfonyl chloride to form the requisite sulfonamide. Physical data as follows: IR (mull) 1745, 1668, 1603, 1535, 1515, 1432, 1413, 1327, 1219, 1179, 1142, 1108, 1073, 1016, 616 cm 'H NMR (CDCI 3 8 8.76 (1 8.17 (2 7.90 (1 7.51 (2 7.32 (4 7.17 (2 5.18 (1 4.96 (1 4.66 (1 4.31 (1 3.78 (3 H), 3.52 (1 3.15 (3 3 C NMR (CDCI.I) 8 171.3. 168.4. 162.2, 147.2, 136.2, 136.0, 132.4, 131.0, 130.3. 128.2. 123.0, 120.4. 120.3, 65.7. 53.6. 52.5, 51.4, 37.3, 34.0; MS (ESI+) for C 27 H ,,C1,F 3 N,0 6 S2 m/lz 690.8 MS (ESI+) for C,.
7
H,-
3 C1 2
F
3
N
4 0 6
S
2 m/Z 712.9 Anal. Calcd for C 27
H
23
C,F
3
N
4 0 6 S. 0.2 H,O: C. 46.68; H, 3.39; N, 8.06.
Found: C. 46.60; H, 3.52; N, 7.92. Water 0.47.
Example rfurmty)2 pyridinyllsulfonyl]-4-thiazolidinyl]carbonylpL-phenylalaine (Scheme A, A-8: where and RA.
2 are the same and equal to H, R 3 is triflouromethylpyridyl), Y is SO 2 R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 S 0 N OH C1
CF
3 Example 45 was prepared from example 44 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1740. 1666, 1602, 1562, 1533, 1517, 1432, 1354, 1327, 1179, 1143, 1108, 1074, 1016. 613 H NMR (DMSO-d 6 8 10.63 (1 9.24 (1 8.54 (1 8.46 (1 8.18 (1 7.50 (5 7.17 (2 5.00 (1 4.74 (1 H), 4.42 (2 3.04 (2 2.90 (1 2.78 (1 3 C NMR (DMSO-d 6 8 172.8, 168.9, 162.3, 159.0. 147.8, 137.5, 137.4, 136.9, 133.5, 131.8, 131.7, 130.2, 128.7, 123.8, 119.7, 64.7, 53.9. 52.3, 36.7, 35.1; MS (ESI+) for C-,H 21
CI
2
F
3
N
4 0 6 ,S m/z 676.5 MS (ESI-) for
C,
6 H,,C12F 3
N
4 06S 2 m/z 674.5 Anal. Calcd for C 26
H
21 C2FN 4 0 6 S, 0.33: C, 45.69; H, 3.20: N, 8.20. Found: C, 45.81: H, 3.38; N, 8.13. Water 0.88.
Example 46.
4 2 ,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-(phenylsulfonyl)-4thiazolidinyl]carbonyl]-L-phenylalanine methyl ester (Scheme A, A-7: where RA^. and RA 2 are the same and equal to H, R 3 is phenyl, Y is SO 2 R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is s- H o O=S=O 0 0 Cl Example 46 was prepared as described in Scheme A from D-cysteine using benzenesulfonyl chloride to form the requisite sulfonamide. Physical data as follows: IR (mull) 1744, 1668, 1604, 1531, 1515. 1432, 1413, 1355, 1324, 1268. 1220. 1195. 1167, 1090. 730 cm 'H NMR (CDC.) 8 7.84 (2 7.65 (5 7.45 (1 7.30 (6 4.90 (1 4.63 (2 4.37 (1 3.75 (3 3.32 (1 3.15 (2 2.53 (1 'C NMR WO 99/67230 PCT/US99/14233 -71-
(CDCI
3 171.2, 168.2. 162.4, 136.4, 136.3. 134.1. 132.4, 131.0, 130.2, 129.6. 128.2, 128.1, 127.9. 120.6. 65.3, 53.3, 52.6, 51.8. 37.4. 33.3: MS (ESI+) for C, 7 ,H,,CI,N3O 6 S, m/z 621.8 MS (ESI-) for C,,H,Cl 2
N
3 0 6 S, mn/z 619.8 Anal. Calcd for
C
27
H
25
CN
3 0 6 S, 0.2 H,O: C, 51.84; H, 4.18; N, 6.72. Found: C, 51.72; H, 4.18: N. 6.52.
Water 0.48.
Example 47.
4 2 6 -Dichlorobenzoyl)amino-N-[[(4S)-3-(phenysulfonyl)-4thiazolidinyl]carbonyl]-L-phenylalanine (Scheme A. A-8: where RA-, and RA- 2 are the same and equal to H. R, is phenyl. Y is SO,, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is NN OH o=S=o N
CI
Example 47 was prepared from example 46 by the procedure described in preparation 6.
Physical data as follows: IR(mull) 1735, 1666, 1605, 1562, 1533, 1516, 1432, 1414, 1352, 1328. 1195. 1180, 1167, 1090, 731 cm'; 'H NMR (CDC 3 8 8.11 (1 7.82 (2 H), 7.66 (3 7.56 (2 7.23 (6 4.91 (1 4.66 (1 4.60 (1 4.35 (1 3.30 (1 3.19 (2 2.59 (1 "C NMR (DMSO-d)8 172.2, 168.2, 161.7, 137.0, 136.9, 136.3, 133.6, 133.0, 131.2, 131.1, 129.6, 129.3, 128.1, 127.6, 119.2, 63.9, 53.4, 51.5, 48.4, 36.0, 33.8; MS (ESI+) for C 26
H
23 Cl2N 3 0 6
S
2 m/z 607.9 MS (FAB) m/z (rel.
intensity) 608 85), 610 608 466 371 228 193 149 129 118(99), 63 HRMS (FAB) calcd for C 2 6
H,
3 C1,N 3 0 6
S
2 608.0483, found 608.0491: Anal. Calcd for C, 6
H
2 3
C,N
3 0 6 S 0.27 H0O: C, 50.91; H, 3.87; N. 6.85.
Found: C, 50.68; H, 4.05; N, 6.65. Water 0.79.
Example 48.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-l[5-(dimethylamino)-lnaphthalenyl]sulfonyl]-4-thiazolidinyl]carbonyl]-L-phenylalanine methyl ester WO 99/67230 WO 9967230PCTIUS99/1 4233 -72- (Scheme A, A-7: where and RA-2 are the same and equal to H. R3 is l -naphthyl, Y is SO 2 R, is 4-[(2,6-dichlorobenzovl)aminojphenvl and stereochemistry is
(SAS).
O=S=O 0
C
N HC6
H
3 C' CH 3 Example 48 was prepared as described in Scheme A from D-cysteine using dimethylamino- 1 -napthalenesulfonyl chloride to form the requisite sulfonamnide. Physical data as follows: IR (mull) 1744, 1684, 1605, 1562, 1533, 1515. 1431. 1412, 1350, 1324, 1231, 1202, 1163, 1145, 798 cm'; 'H NMR (CDC 3 8 8.64 (1 8.34 (2 7.53 (5 H), 7.29 (4 6.87(3 4.93 (1 4.75 (1 4.64 (1 4.31 (1 3.69(3 3.47 (1 2.84 (8 2.46 (1 3 C NMR (CDC 3 5 171.2, 167.7, 162.5, 136.3, 135.9, 132.4, 132.3, 131.9, 131.5, 131.0, 130.1, 129.7, 129.2, 128.2, 124.4, 120.4, 65.1, 53.3, 52.5, 50.1, 45.9, 37.1, 33.3, MS (ESI+) for C 3 3
H
32 C1 2
N
4 0 6 m/z 736.8 Anal.
Calcd for C 33
H
32 Cl 2
N
4 0 6 S, 0. 17 H,O: C. 55.15; H, 4.54: N, 7.79. Found: C 55.20; H, 4.73; N, 7.49. Water 0.43.
Example 49.
4 -II(2,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-[[5-(dimethylamino)-I naphthalenyljsulfonyl]-4-thiazolidinyljcarbonyl]-L-phenylalanine (Scheme A, A-8: where and RA- 2 are the same and equal to H. is 1 -naphthyl, Y is SO, R, is 4-[(2,6-dichlorobenzoyl)aminolphenyl and stereochemnistry is
S))
WO 99/67230 PCT/US99/14233 -73- NY OH o=S=o0 O0 C
N
H
3 C' 'CH 3 Example 49 was prepared from example 48 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1666, 1605. 1587, 1577, 1562, 1532, 1516, 1431, 1412, 1395, 1325, 1163, 1145, 798, 631 cm 'H NMR (DMSO-d 6 6 10.60 (1 8.54 (1 8.28 (2 7.54 (8 7.25 (1 6.93 (2 4.95 (1 4.84 (1 4.39 (1 H), 4.17 (1 2.95 (2 2.80 (7 2.54 (1 3 C NMR (DMSO-d 6 8 167.2, 161.7, 151.4, 136.6. 136.3, 133.6, 133.1, 131.1, 130.8. 130.3, 129.5, 129.3, 129.0, 128.8, 128.1, 123.6, 118.9, 118.3, 115.3, 63.6, 54.2, 50.3, 44.9. 36.7, 33.9, 21.0; MS (ESI+) for
C
3 2
H
3 0 C1,N 4 0 6
S
2 m/z 700.8 HRMS (FAB) calcd for C 3 2 H3o 0
CIN
4 06S 2
+H,
701.1062, found 701.1039 Example 2 ,6-Dichlorophenyl)methyl]-N-[[(4S)-3-(methylsulfonyl)-4thiazolidinyl]carbonyl]-L-tyrosine methyl ester (Scheme A. A-7: where RA.- and RA-2 are the same and equal to H, R 3 is methyl, Y is SO 2 R, is 4 2 ,6-dichlorophenyl)methoxy]phenyl and stereochemistry is o=S=o 0 I CCH3 Example 50 was prepared as described in Scheme A from D-cysteine using methanesulfonyl chloride to form the requisite sulfonamide. Physical data as follows: IR (mull) 1742. 1680, 1611, 1564, 1510. 1439, 1345. 1299, 1240, 1179, 1158, 1016, 976, 779, 768 'H NMR (CDCI 3 6 7.36 (2 7.24 (1 7.08 (3 6.97 (2 5.25 (2 4.77 (3 4.29 (1 3.74 (3 3.43 (1 3.53 (1 3.10 (2 2.93 (3 H); WO 99/67230 WO 9967230PCTIUS99/1 4233 -74- 3 C NMR (CDCI) 6 171.4, 168.0. 158.2. 137.0, 132.1. 130.3. 1218.5, 127.9, 115.3. 65.2, 59.4, 53.5, 53.3. 52.5. 51.9. 42.2, 37.3, 36.9, 34.2: MIS (ESI+) for C, H 4 m/z 546.8 MIS (ESI+) for C 22
H
24 C1IN 2 0 6 m/z 568.8 HRMS (El) calcd for C 22
H,
4 C1., 2 0 6 546.0453, found 546.0448; Anal. Calcd for C,,H 24 C1,N.,0 6
S
2 -0.07 H,O: C. 48.15; H, 4.43; N, 5. 10. Found: C, 48.17; H. 4.5 1; N. 5.02. Water 0.24.
Example 51.
2 6 -Dichlorophenyl)methyl]-N-[[(4S-3 (methylsulfbnyl)-4thiazolidinyl]carbonyl]-L-tyrosine (Scheme A, A-8: where RA-I and RA- 2 are the same and equal to H, R 3 is methyl, Y is SO, R, is 4 2 6 -dichlorophenvl)methoxyjphenyI and stereochemistry is
-OH
0
CI
C4 Example 51 was prepared from example 50 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1737, 1675, 1611, 1565. 1511, 1439, 1345, 1300, 1241, 1197.,1179, 1157.,1016, 778, 769 'H NMR (CDCl 3 867.34 (2 7.16 (5 H), 6.99 (2 5.23 (2 4.85 (1 4.68 (2 4.27 (1 3.51 (1 3.32 (1 3.15 (2 2.93 (3 3 C NMR (CDCl 3 6 174.8, 168.7, 158.3. 137.0, 132.1, 130.5, 130.4, 128.5, 127.6, 115.4, 115.2.65.2, 64.9, 53.1, 52.0, 37.1. 36.4, 3 4.3; MIS (ESI+) for
C
2 ,H,,C1 2 N,0 6 S. m/z 532.8 MIS (ESI-) for C, 1
H
22 C1,N, 2 0 6
S
2 mn/z 530.7 HRMS (FAB) calcd for C,,H,,C1,N 2
O
6
S
2 533.0374, found 533.0386; Anal. Calcd for
C
21
H
22 ,C1,N,0 6 5, 0.06 H,O: C. 47.19; H, 4.17; N. 5.24. Found: C. 47.58; H, 4.35; N, 10. Water 0.20.
Example 52.
4-[(2,6-Dichlorobenzovl)amino]-N-[[(4Sy..3{[[( 1.1-dimethylethvl )amino]carbonyl]-4thiazolidinyl]carbonylpL-phenvlalanine WO 99/67230 PCTIUS99/14233 (Scheme A. A-8: where RA-1 and RA 2are the same and equal to H. R, is t-butyl, Y is CONH-, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is N OH N O 0 0 C1 H N Example 52 was prepared from example 6 by the procedure described in preparation 6.
Physical properties as follows: IR (mull) 3289. 1728. 1664. 1607. 1580. 1561. 1536, 1432, 1414, 1394, 1326, 1270, 1242, 1213. 1195 NMR (300 MHz, CD 3 0D) 8 7.47 (2 7.24 (3 7.04 (2 4.63 (2 4.37 (1 4.17(1 3.09 (4 1.22 (9 "C NMR (75 MHz, CDCI 3 8 176.9, 174.8, 167.2, 159.9, 140.5, 140.0, 136.4, 136.1, 134.6, 133.8, 131.8, 124.5, 66.6, 57.1, 55.3, 53.0, 40.1, 37.1, 32.9; MS (ESI+) for C2, 5 H, CN 4 0S m/z 566.9 588.9 MS (ESI+) for C,,H 28 CIN40OS m/z 566.9 HRMS (FAB) calcd for C2,H2, 8
CL,N
4 0,S +Hl 567.1235, found 567.1253.
Example 53.
4 2 6 -Dichlorobenzoyl)amino]-N-[[(4S)-3-[(diethylamino)carbonyl]-4thiazolidinyl]carbonyl]-L-phenylalanine (Scheme A, A-8: where RA. and are the same and equal to H, R, is ethyl, Y is
CON(CH,CH
3 R, is 4 2 6 -dichlorobenzoyl)amino] phenyl and stereochemistry is (S, S HO N OH ^N o 0 0 o ci Example 53 was prepared from example 7 by the procedure described in preparation 6.
Physical properties as follows: IR (mull) 3269, 1734, 1663, 1607. 1562. 1535, 1515, WO 99/67230 PCT/US99/14233 -76- 1431, 1415. 1348. 1325. 1269, 1213, 1195, 799 cm'; 'H NMR (300 MHz, CD 3 OD) 8 7.57 (2 7.35 (3 7.09 (2 5.09(1 4.76(1 4.38 (2 3.31 (3 3.13 (5 H), 1.05 (6 NMR (75 MHz, CD30D) 8 176.6. 173.9. 167.1, 166.3, 140.7, 140.1, 136.1, 134.6, 133.8. 131.8, 124.1, 68.5, 57.4, 56.6, 45.9. 40.4, 36.6. 16.8; MS (ESI+) for
C
25
H
28
C
2
N
4 0,S m/z 567.1 Anal. Calcd for C, 5
H,
8 C1 2
N
4 0 5 S: C, 52.91; H, 4.97; N, 9.87.
Found: C, 52.60; H. 5.13; N, 9.47.
Example 54.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(4S)-3-[[methyl pyridinyl)ethyl]amino]carbonyl]-4-thiazolidinvl]carbonyl ]-L-phenylalanine methyl ester (Scheme A, A-7: where RA-1, and RA 2 are the same and equal to H, R 3 is 2-(2-pyridl)ethyl, Y is CON(CH 3 R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (S, NNO N O O CI HcI o Example 54 was prepared as described in Scheme A from D-cysteine using 2-(2methylaminoethyl)pyridine to form the requisite urea. Physical data as follows: mp (dec); IR (mull) 1743, 1665, 1606, 1561. 1538, 1514, 1489, 1432, 1413, 1395, 1323, 1268, 1216, 1195, 799 cm 'H NMR (300 MHz, DMSO-d 6 6 8.44 (1 8.34 (1 H), 7.66 (1 7.51 (5 7.19 (4 4.72 (1 4.48(1 4.40(1 4.20 (1 3.58 (4 3.42 (1 3.89 (10 "C NMR (75 MHz. DMSO-d 6 8 172.1, 170.3, 162.3, 161.8, 159.4. 149.4. 137.6, 137.0, 136.8, 133.3. 131.8. 131.7. 130.0, 128.7, 123.8, 122.0, 119.8, 64.58. 53.8. 52.7. 52.5, 49.7, 36.5, 36.2. 35.7. 33.4; MS (ESI+) for C 30
H
3 1 C1,NO 5
S
m/z 643.9(M+H)-; Anal. Calcd for C 3 0
H
3 ,C1,N 5 0,S: C. 55.90: H, 4.85; N, 10.86. Found: C, 55.52; H. 5.09; N. 10.64.
WO 99/67230 WO 9967230PCTIUS99/ 14233 -77- Example 4-[(2,6-Dichlorobenzoyl)aminol-N-[[(4S)-3-[methvl pyridinyl)ethyl]amino]carbonyl]-4-thiazolidinyllcarbonyl]-L-phenylalanine (Scheme A, A-8: where RA., and RA.
2 are the same and equal to H. is 2-(2-pyridyl)ethyl, Y is CON(CH 3 R, is 4-[(2,6-dichlorobenzoyl)amino]phenvl and stereochemnistry is (S, N N '-O 0 0 0 C1 Example 55 was prepared from example 54 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1682, 1656, 1606, 1561, 1540, 1513, 1432, 1413, 1398, 1323, 1268, 1242, 1195. 799,780 cm 'H NMR (300 MHz, DMS0-l 6 5 10.66 (1 8.45 (1 8.09 (1 7.66 (1 7.51 (5 7.20 (4 4.72 (1 4.40 (2 H), 4.23 (1 3.61 (1 3.40 (1 3.04 (7 2.79 (3 MS (ESI+) for C, 9
H,
9 C 1 2
N
5 0 5 m/z 629.9 Anal. Calcd for C, 9
H
29 C1 2
N
5 0 5 S 0.61 1-LO: C. 54.29; H. 4.75; N, 10.92. Found: C, 54.29; H, 5.00: N, 10.32. Water 1.72.
Example 56.
4 -[(2,6-Dichlorobenzovl)amino]-N-[[(4S)-3-(4-morpholinylcarbonyl)-4thiazolidinyl]carbonyl]-L-phenylalanine methyl ester (Scheme A, A-7: where and RA.
2 are the same and equal to H. R~ and Y together form CO-morpholino, R, is 4-[(2.6-dichlorobenzoyl)amino]phenyl and stereochemnistry is (S, WO 99/67230 WO 9967230PCTIUS99/I 4233 -78s- 0 Hi HCI6 Example 56 was prepared as described in Scheme A from D-cysteine using morpholine to form the requisite urea. Physical data as follows: mp 223-225-C; I HNMR (300 MHz, DMSO-1 6 6 10.66 (1 8.25 (1 7.51 (5 7.16 (2 4.81 (1 4.60 (1 4.50 (1 4.28 (1 3.64 (3 3.53 (4 3.09 (8 3 C NMR (75 MHz, DMSO-d 6 172.1. 170.3. 162.3, 161.4, 137.6. 136.8. 133.3. 131.8. 131.7, 130.0, 128.7. 119.8. 66.2, 64.3, 53.8. 52.8, 52.5, 46.8, 36.1, 33.7; HRMS (FAB) calcd for C,- 6
H
2 ,CL,N,0 6
S+H
1 595.1185, found 595.1189: Anal. Calcd for C 26
H
2 8
C,N
4 0 6 S: C, 52.44; H, 4.74; N. 9.41.
Found: C. 52.42; H, 4.96; N, 9.23.
Example 57.
4
S)-
3 -[[Bis(2-hydroxyethyl)amino]carbonyly4thiazolidiny llcarbonvl]-4+[2,6dichlorobenzovl)amino]-L-phenylalanine methyl ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to H, R 3 is 2-hydroxyethyl and Y is CON(CHCHOH). R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemnistry is S.
HI
HO.~H
Example 57 was prepared as described in Scheme A from D-cysteine using diethanolamine to form the requisite urea. Physical data as follows: mp 105-107"C; IR (mull) 3284. 1743. 1662. 1608, 1561, 1539. 1516, 1432, 1414, 1355, 1326, 1270. 1217, 1196.,799 cm NMR (300 MHz. DMSO-d,) 8 10.69 (1 8.28 (1 7.56 (4 H), WO 99/67230 PCT/US99/14233 -79- 7.47 (1 7.16 (2 4.87 (3 4.66 (1 4.47(1 4.25 (1 3.63 (3 3.47 (6 3.02 (6 MS (ESI+) for C, 6
H
3 0
C
2
N
4 0,,S m/z 612.9 Anal. Calcd for
C,,H
30 C1 2
N
4 0,S 0.47 HO: C, 50.21; H, 5.01: N, 9.01. Found: C, 50.02; H. 5.00; N, 8.93.
Water(KF): 1.36.
Example 58.
4 S)-3-[[Bis(2-hydroxyethy)amino]carbonyl]-4-thiazolidinylcarbonyl]-4-[(2,6dichlorobenzoyl)amino]-L-phenylalanine (Scheme A, A-8: where RAI and RA- 2 are the same and equal to H, R, is 2-hydroxyethyl and Y is CON(CHCH,OH), R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is 0
H'-O
H CI6 Example 58 was prepared from example 57 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3281, 3196, 1724, 1660, 1608, 1580, 1561, 1542, 1515, 1431. 1415, 1354. 1328, 1271, 1196 cm 1 H NMR (300 MHz, DMSO-d, 6 8 10.65 (1 8.13 (1 7.55 (4 7.47 (1 7.16 (2 4.85 (2 4.66 (1 4.40 (1 H), 4.27 (1 3.48 (6 3.01 (6 MS (ESI+) for CH 28
CI,N
4 0S m/z 598.9 Anal. Calcd for C 2 5H, 8
C,N
4 0S 1.04 HzO: C, 58.58; H, 4.90; N, 9.06. Found: C, 48.88; H, 5.05; N, 8.79. Water 3.02.
Example 59.
4 2 6 -dihlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-&-oxo-3-thiazolidinepentanoic acid 3-methyl ester (Scheme A, A-7: where RA.1 and RA-2, are the same and equal to H. R, is
CH
1
CH,CH,CO,CH
3 Y is CO, RS is 4-[(2,6-dichlorobenzoyl)amino]phenvl, and stereochemistry is WO 99/67230 PCT/US99/14233 o O 6 Example 59 was prepared as described in Scheme A from D-cysteine using methyl glutaryl chloride to form the requisite amide. Physical data as follows: IR (mull) 3266, 1741, 1734, 1685, 1678, 1630, 1610. 1560, 1545, 1441, 1435, 1414, 1327, 1268, 1227 cm 'H NMR (DMSO-d 6 8 10.65 (1 8.43 (1 7.50 (5 7.15 (2 4.72 (2 4.44 (2 3.63 (3 3.56 (3 3.10 (4 2.15 (4 1.70 (2 "C NMR (CDC1 3 8 171.6. 170.6, 170.1, 169.9, 169.6, 161.9, 137.2, 136.5, 132.9, 131.3, 129.7, 128.3, 119.4, 61.4, 53.7, 53.5, 52.0, 51.3, 48.7, 36.4, 35.8, 35.1, 33.1, 32.8, 32.5, 25.4, 19.8; MS (ESI+) for C2, H 9
N
3 0,SCI 2 m/z 610.0 MS (ESI-) for C,,H2 9
N
3 0,SCI, m/z 608.0 Anal. Calcd for C2,H, 9
CI
2
N
3 0 7 S: C, 53.12; H, 4.79; N, 6.88. Found: C. 52.81; H, 4.90; N, 6.88 Example dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-Z-oxo--3-thiazolidinepentanoic acid (Scheme A, A-8: where RA^. and RA.2 are the same and equal to H, R, is
CHCH,CH,CO
2 H, Y is CO, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is s- o 0O N -OH HO 0 0 CI Example 60 was prepared from example 59 by the procedure described in preparation 12.
Physical data as follows: IR (mull) 3271. 3193, 3124, 1725. 1661, 1607. 1561. 1539.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -81- 1516, 1432, 1414. 1327, 1271, 1195, 799 cm -1 HNR MSd 6 8 12.40 (1 10.63 (1 8.31 (1 7.50 (5 7.16 (2 4.75 (2 4.34 (2 2.95 (4 2.15 (4 H), 1.66 (2 3 CNMR (CD3CN) 8 175.0, 172.7, 172.6. 171.0. 163.5, 137.8, 13 7.0. 134.4.
132.7, 132.3, 131.1, 130.9, 129.1. 120.7. 63.2, 54.5. 49.9. 37.2. 34.1, 33.3, 20.7; MS, (ESI+) for C,SH,SC1,N 3 0 7 S m/z 582.0 (M+H) t MS (ESI-) for C, 5
H
25 C1,N 3 0 7 S m/z 579.9 Anal. Calcd for C., 5
H
25 Cl,N 3 0.
7 S C. 50.01; H. 4.53; N, 7.00. Found: C, 49.61; H, 4.38; N. 6.61.
Example 6 1.
-Acetyl-4-thiazolidinyl]carbonyl]-4-(2,6-dichlorobenzoyl)amino-Lphenylalanine, methyl ester (Scheme A, A-7: where RA-.I and RA.
2 are the same and equal to H. R 3 is CH 3 Y is CO, R is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is
HI
Example 61 was prepared as described in Scheme A from D-cysteine using acetyl chloride to form the requisite amide. Physical data as follows: IR (mull) 3260, 3067, 1748, 1686, 1623, 1608, 1561. 1542, 1515, 1445, 1429,1419,1324.1267,1221 'H NMR (DMSO-d 6 5 8.44 (1 7.52 (5 7.17 (2 4.78 (2 4.37 (2 3.63 (3 3.06 (4 1.94 (3 3 C NMR (DMF-d,) 8 172.4, 170.6. 138.4. 137.5. 133.8, 132.3. 131.9, 130.6, 128.9, 120.0. 70.9, 63.6, 62.4, 54.7, 52.3. 50.2. 49.8. 49.2, 37.2, 36.6, 22.7; MS for C 23
H.,
3 C1 2 N.10 5 S m/z 523.9 MS (ESI+) for C 23
H
23 C1., 3 0 S ,nz 545.8 HRMS (FAB) calcd for C2 3
H,-
3 C1,-N 3 0 5 S+H, 524.0814, found 524.0812; Anal.
Calcd for C', 3
H
23 C1..N 3
O
5 S 1 H,O: C, 52.46; H. 4.45; N, 7.98. Found: C, 52.85; H, 4.42; N, 8.00. Water 0.24.
WO 99/67230 PCT/US99/14233 -82- Example 62.
N-[[(4S)-3-Acetyl-4-thiazolidinyl]carbonyl]-4-[(2,6-dichlorobenzoyl)amino]-Lphenylalanine (Scheme A, A-8: where RA, and RA-2 are the same and equal to H. R, is CH 3 Y is CO, R3 is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is 0 N OH ooC N I c Example 62 was prepared from example 61 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3070, 1747, 1682, 1663, 1625. 1608, 1580, 1561, 1548, 1514, 1443, 1431, 1416,1278, 1220 cm 'H NMR (DMSO-d6) 5 12.48 (1 H), 10.63 (1 8.29 (1 7.50 (5 4.73 (2 4.34 (2 2.97 (4 1.93 (3 H); 1 3 C NMR (DMSO-d 6 )8 172.4, 169.6, 169.2,168.5, 168.0,161.7, 136.9, 136.3, 133.2, 133.0, 131.2, 129.6, 128.1, 119.2, 62.2, 61.0, 53.5, 53.3, 49.3, 48.4, 36.3, 35.7, 35.0, 33.1, 22.4, 20.9; MS (ESI-) for C, 2
H
2 ,,ClN 3 0SS m/z 507.9 Anal. Calcd for C 2H,,C1,N 3 0S 0.1 H 2 0,O: C, 51.57: H. 4.18; N, 8.20. Found: C. 51.49: H, 4.36; N, 8.07.
Water 0.40.
Example 63.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-5,5-dimethyl-6-oxo-3thiazolidinebutanoic acid (Scheme A, A-8: where RA- and RA- 2 are the same and equal to CH., R 3 is CH,CH 2
CO
2
H,
Y is CO. R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is WO 99/67230 PCT/US99/14233 -83-
HO
Hao Example 63 was prepared as described in Scheme A from D-penicillamine using methyl succinyl chloride to form the requisite amide. Physical data as follows: IR (mull) 3264, 3198, 3071, 1721, 1660, 1608, 1562, 1541, 1516, 1432, 1415, 1327, 1270, 1195, 799 cm'; 'H NMR (DMSO-d 6 6 10.62 (1 8.15 (1 7.52 (5 7.18 (2 4.72 (2 4.43 (3 2.91 (4 2.17(2 1.36(3 1.07(3 "C NMR (DMSO-d 6 6 173.5, 172.6, 170.2, 169.7, 168.4. 168.1, 161.7, 136.9, 136.3, 133.3, 131.1, 129.6, 129.4, 128.1, 119.2, 70.6, 53.8, 53.5, 51.6, 48.4, 47.7, 35.9, 30.7. 30.5, 28.9, 28.6. 24.1; MS (ESI+) for
C
2 6
H,
7 C1,N 3 0 7 ,S m/z 596.0 MS (ESI+) for C 26
H,
7 C1,N 3 0,S m/z 617.9 MS (ESI-) for C 6
H
27 C1,N30, 7 S m/z 593.8 MS (FAB) m/z (rel. intensity) 596 20), 598 596 331 193 141 139 116(16), 107 (13), 105 89 HRMS (FAB) calcd for C, 6
H
27
C,N
3 0 7 S+HI 596.1025, found 596.1036.
Example 64.
-Carboxy-2-[4-[(2,6dichlorophenyl)methoxyjphenyl]ethyl]amino]carbonyl]-y-oxo-3-thiazolidinebutanoic acid (Scheme A, A-8: where RA-1, and RA- 2 are the same and equal to H, R 3 is CH 2
CH,CO
2 H, Y is CO, R, is 4-[(2,6-dichlorophenyl)methoxylphenyl, and stereochemistry is s- o H O (N'r -'OH HO 00c "lo
N
cib Example 64 was prepared as described in Scheme A from D-cysteine using methyl succinyl chloride to form the requisite amide. Physical data as follows: IR (mull) 3073, WO 99/67230 PCT/US99/14233 -84- 3031, 1725, 1640, 1612. 1585. 1565, 1535, 1511, 1439, 1300. 1241, 1196, 1179, 768 cm-'; 'H NMR (DMSO-d 6 8 8.26 (1 7.53 (2 7.44 (1 7.12 (2 6.93 (2 5.16 (2 4.75 (2 4.40 (2 4.20 (1 2.81 (4 2.32 (2 2.07 (1 3C NMR (DMSO-d 6 8 173.8, 172.6, 170.0, 169.3, 157.1, 136.0, 131.5, 130.4. 130.0, 129.8, 128.7, 114.2, 64.8, 61.5, 53.6, 48.5. 36.1. 35.5, 35.0, 33.1, 28.9, 28.6. 21.0; MS (ESI+) for
C
2 4
H
24 C1 2
N
2 0,S m/z 554.8 MS (ESI-) for C 24
H
24 CI,N20,S m/z 552.7 HRMS (FAB) calcd for C 24 H 4 CI2N,0 7 S+H, 555.0759, found 555.0750.
Example 4
S)-
3 -Acetyl--thiazolidinyl]carbonyl]-O-[(2,6-dichlorophenyl)methyl]-Ltyrosine methyl ester (Scheme A, A-7: where RA., and RA.
2 are the same and equal to H, R, is CH 3 Y is CO, R is 4-[(2,6-dichlorophenyl)methoxy]phenyl, and stereochemistry is o N. O H3C- 0 o Cl Example 65 was prepared as described in Scheme A from D-cysteine using acetyl chloride to form the requisite amide. Physical data as follows: IR (mull) 1744, 1657, 1612, 1585, 1564, 1511, 1438, 1405, 1352, 1299, 1240, 1197, 1179, 1016, 768 cm'; 'H NMR (CDCl 3 8 7.36 (2 7.24 (1 6.97 (5 5.24 (2 5.04 (1 4.78 (1 4.50 (2 3.74 (3 3.45 (1 3.17 (3 2.02 (3 "C NMR (CDC13) 8 172.7, 171.6, 171.4, 169.8, 168.9, 168.7, 157.8, 136.9, 131.9, 130.3, 129.9, 128.6, 128.4, 114.9, 65.1, 61.6, 56.0, 53.6, 53.3, 49.6, 36.7, 31.7, 22.5; MS (ESI+) for C 2 3
H
24 C1 2
N
2 0,S m/z 532.9 MS (EI) m/z(rel. intensity) 510 338 337 336 267 (12 265 163 161 159 130 88 Anal. Calcd for C 23
H,
4 C1,NOS*.0.19 H,O: C, 53.66; H, 4.77; N, 5.44. Found: C. 53.81; H, 4.75; N, 5.33. Water 0.66.
WO 99/67230 PCT/US99/14233 Example 66.
4
S)-
3 -Acetyl--4-thiazolidinyl]carbonyl]-O-[(2,6-dichlorophenyl)methyl]-Ltyrosine (Scheme A, A-8: where and RA- 2 are the same and equal to H, R, is CH 3 Y is CO, R is 4 -[(2,6-dichlorophenyl)methoxy]phenyl, and stereochemistry is s- H 0 N OH
H
3 CO 0 0 Example 66 was prepared from example 65 by the procedure described in preparation 6.
Physical data as follows: IR(mull) 1730, 1646, 1612, 1585, 1565, 1511, 1439, 1414, 1299, 1240, 1196, 1179, 1016, 779, 768 cm'; 'H NMR (DMSO-d 6 6 8.26 (1 7.54 (2 7.44 (1 7.13 (2 6.92 (2 5.16 (2 4.72 (2 4.31 (2 2.91 (5 H), 1.92 (3 "C NMR (DMSO-d 6 8 172.6 169.2, 168.0, 157.1, 136.0, 131.7, 131.5, 130.3, 128.7, 114.2, 64.8, 62.2, 61.0, 53.7, 49.4, 48.5, 36.1, 35.5, 35.1, 33.2, 22.4, 21.0; MS (ESI+) for C,2H 22
CI,N
2 0 5 S m/z 496.9 MS (ESI-) for C 22
H
22 Cl 2
N
2 0,S m/z 494.8 MS (FAB) m/z (rel. intensity) 497 99), 617 573 539 500( 11), 499 498 497 496 225 130 HRMS (FAB) calcd for 5 S+H, 497.0705, found 497.0713; Anal. Calcd for C 2 2
,H,
2 C1,N,O0 5 S 0.41
H
2 0: C. 52.35; H, 4.56; N, 5.55. Found: C, 52.65; H, 4.51; N, 5.50. Water 1.46.
Example 67.
1 -Carboxy-2-[4-[(2,6dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-y-oxo-3-thiazolidinebutanoic acid (Scheme A, A-8: where RA,1 and RA-2, are the same and equal to H, R 3 is CH,CH,COH, Y is CO, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is WO 99/67230 PCTIUS99/14233 -86- SOH
CI
Example 67 was prepared as described in Scheme A from L-cysteine using methyl succinyl chloride to form the requisite amide. Physical data as follows: 'H NMR
(CD
3 CN) 5 8.86 (1 7.55 (2 7.42 (3 7.24 (2 7.11 (1 4.90 (1 4.65 (2 4.33 (1 3.14 (3 2.47 (6 1.80 (1 3 C NMR (CD 3 CN) 8 174.7, 172.6, 172.3, 170.7, 163.4. 137.7, 137.0, 134.4, 132.7, 132.2, 130.9. 129.1, 63.2, 60.9, 54.4, 49.7, 37.1, 36.7, 32.7, 30.6, 30.0, 29.5, 21.1, 14.4; MS (FAB) m/z (rel. intensity) 568 (MH+, 99), 646 644 572 571 570 569 568 567 216 88 Anal. Calcd for C,, 4
H
2 3 ClN 3 0 7 S 0.5 HO: C. 49.92; H, 4.19; N, 7.23.
Found: C, 50.01; H, 4.54; N, 7.05.
Example 68.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]--oxo-3-thiazolidinepentanoic acid (Scheme A, A-8: where RA., and are the same and equal to H, R 3 is
CH
2
CHCH
2 COH, Y is CO, R 5 is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is
OHOH
c Example 67 was prepared as described in Scheme A from L-cysteine using methyl glutaryl chloride to form the requisite amide. Physical data as follows: 'H NMR (CD 3 CN) 8 8.85 (1 7.55 (2 7.44 (3 7.17 (3 4.86 (1 4.64 (2 4.34 (1 3.64 (1 H), WO 99/67230 PCT/US99/14233 -87- 3.13 (3 2.27 (4 1.79 (4 3 C NMR (CD 3 CN) 8 175.0, 172.9. 172.7. 170.6, 163.4, 137.7. 137.0, 134.2, 132.7, 132.2, 131.0. 129.1, 120.7, 68.2, 63.1, 54.2. 49.8. 37.1, 36.9, 34.0. 33.2, 32.6, 26.2, 20.6; MS (FAB) m/z (rel. intensity) 582 99), 585 584 583 582 99), 581 88 69 57 55 43 Anal. Calcd for C2,H, 5 C1N 3 0,S 0.2 HO 2 0: C, 51.24; H, 4.37; N, 7.17. Found: C, 51.25; H, 4.68; N, 6.92.
Example 69.
4
R)-
3 -Acetyl-4-thiazolidinyl]carbonyl]-4-[(2,6-dichlorobenzoyl)amino]-Lphenylalanine methyl ester (Scheme A, A-7: where RA,- and RA 2 are the same and equal to H, R, is CH 3 Y is CO, R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is H 3 C O
C
0 Cl6 Example 69 was prepared as described in Scheme A from L-cysteine using acetyl chloride to form the requisite amide. Physical data as follows: IR (mull) 3268, 1743, 1662, 1607, 1561, 1538, 1515, 1431, 1413, 1354, 1324. 1270, 1217, 1195, 799 NMR (CDC 3 6 7.56 (3 7.31 (4 7.14 (2 4.92 (2 4.49 (1 4.29 (1 3.77 (3 3.54 (1 3.26 (1 3.00 (2 2.11 (3 3 C NMR (CDCI 3 6 171.5, 170.3, 168.9, 152.5, 135.9, 135.7, 132.9, 132.3, 131.0, 130.1, 129.9, 128.2, 128.0, 127.9, 120.4, 63.9, 61.4, 53.1, 52.5, 49.7, 37.0, 31.2, 22.6; MS (ESI+) for C 23
H
2 3
C,N
3 0S m/z 523.8 Example N-[[(4R)-3-Acetyl-4-thiazolidinyl]carbonyl]-4-[(2,6--ichlorobenzoyl)amino]-Lphenylalanine (Scheme A, A-8: where RA-, and RA., are the same and equal to H. R 3 is CH,, Y is CO, R is 4 2 ,6-dichlorobenzoyl)amino]phenyi, and stereochemistry is WO 99/67230 PCT/US99/14233 -88- N OH
H
3 C0 o V 0 ci N S'oo was prepared from example 69 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3276, 3068, 1727, 1661, 1607, 1561, 1540, 1516, 1444, 1431, 1414, 1327, 1271, 1218, 1195 'H NMR (DMSO-d 6 12.78 (1 10.67 (1 8.27 (1 7.49 (4 7.19 (2 4.75 (2 4.45 (3 3.01 (3 1.96 (3 H); "C NMR (DMSO-d 6 172.6, 172.3, 169.4, 168.6. 168.2, 161.7, 136.9, 136.3, 133.3.
132.8, 131.1, 129.6, 128.1, 119.1, 62.2, 60.9, 53.4, 49.3, 48.7, 35.9, 35.1, 32.7, 22.4; MS (ESI+) for C,,H 2
,CI,N
3 0 5 S m/z 509.8 MS (ESI-) for C 2 2
H
21 ,ClN 3 0S m/z 507.8 HRMS (FAB) calcd for C2,H 2 1 Cl 2
N
3 0S+H, 510.0657, found 510.0667; Example 71.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(4R)-4-thiazolidinyl]carbonyl]-L-phenylalanine monohydrochloride salt (Scheme A, A-10: where RA.- and RA2 are the same and equal to H, R, is dichlorobenzoyl)amino]phenyl, and stereochemistry is <Won H 0 0 0o c Example 71 was prepared as described in Scheme A from L-cysteine. Physical data as follows: IR (mull) 3249, 3190, 3036, 1729, 1662. 1605, 1578, 1562, 1541, 1516, 1432, 1414, 1328, 1271. 1195 'HNMR (DMSO-d 6 6 10.71 (1 8.83 (1 7.56 (5 H), 7.24 (2 4.50 (1 4.25 (1 4.21 (2 3.62 (1 3.01 (4 3 C NMR (DMSOd 6 8 172.0, 166.8. 161.8, 137.7, 132.7, 131.3, 131.0. 129.5, 128.1, 119.4, 72.0, 70.4. 62.3, 53.8, 49.0, 35.7. 33.4; MS (ESI+) for C, 0 H,,CIlNOS m/z 468.1 WO 99/67230 WO 9967230PCTIUS99/1 4233 -89- Scheme B R-B.2 B-i
H
2 N
OH
0 4 OH B-2 RB -1R 9 4 OHB4
RB-
RB-1 0 R NHU
R
3
RB
5 RB RB4 N yNH RZ RB- I B-7 WO 99/67230 PCT/US99/14233 where R1_, RB.
3 and are defined as R, and in addition R.
3 and may be attached to the same carbon atom and form a cyclic ring of 5-8 atoms of the formula: (CH2) together with the carbon atom to which they are attached; R,.s is defined as OH or O-(C.
6 alkyl) Scheme B describes a general method for the preparation ofthiazolidine-4carboxylic acid derivatives of general structure B-6. B-7 and B-8 which are disubstituted at the 2 position RB.
3 and are not equal to Within this class of structures, the nitrogen is derivatized immediately after forming the heterocyclic ring. Accordingly, a commercially available or readily prepared sulfur-containing amino acid of structure B-1 (which is the same structure as A-i) is condensed with a commercially available or readily prepared ketone to afford the thiazolidine-4-carboxylic acid of general formula B-3 (preparation 15) (for a general discussion of the condensation of aldehydes and ketones with cysteine or similar sulfur containing amino acids see: Coppola, Schuster, H.F.
Asymmetric Synthesis: Construction of Chiral Molecules Using Amino Acids; John Wiley: New York, 1987; Chapter 6, 171.). The amine group may be reacted with a variety of electrophilic reagents such as sulfonyl chlorides, carbonates, chloroformates. isocyanates, phosgene (or a suitable equivalent) and an amine, acid chlorides, and carboxylic acid anhydrides as described in Scheme A for the reaction of A-6. Preparation 16 is provided as a specific example of the synthesis of a compound of general structure B-4.
Condensation of B-4 with amino acid derivative B-5 under standard peptide synthesis conditions provides the compound of general structure B-6 (preparation 17). Mild base hydrolysis of the methyl ester of general structure B-6 (where RB.
5 is OCH 3 may be effected as described and exemplified in Scheme A (preparation 6 or 13) to afford compounds of the general structure B-7. Alternatively, in those cases where RB, is O-t- Bu, mild acidolysis can also provide compounds of the general structure B-7 (by the procedure described in preparation 4 of Scheme In the case of t-butoxycarbonyl derivatised analogs of general structure B-7 where R, is t-butyl and Y is mild WO 99/67230 PCT/US99/14233 -91acidolysis affords compounds of general structure B-8 (by the procedure described in preparation 4 of Scheme A).
Preparation (Scheme B, B-3: where and RB.
2 are the same and equal to H, and RB are the same and equal to CH,. and stereochemistry is H3
C
Qy
H
0 A suspension of L-cysteine hydrochloride monohydrate (Scheme B. B-I: where RB., and
RB.
2 are the same and equal to H and stereochemistry is (20 g, 0.11 mol) in acetone (Scheme B, B-2: where RB 3 and RB 4 are equal to CH 3 (800 mL) was heated to reflux for 8 h. Cooling to room temperature resulted in precipitation of a solid which was collected by filtration, washed with acetone and dried in vacuo to afford the title compound (17.46 g) as a white solid: mp 165-167 0 C; (Lit. 165-168 0 C: Sheehan, Yang, D-D.H. J. Am.
Chem. Soc. 1957, 80, 1158) 'H NMR 5 4.75 (1 3.59 (1 3.44 (1 1.73 (3 1.71 (3 MS (ESI-) for C 6 H,,NOS m/z 159.9 Preparation 16.
(Scheme B, B-4: where RB., and R- 2 are the same and equal to H. R 3 and RB, are the same and equal to CH3, Y is CO 2
R
3 is t-butyl and stereochemistry is 7( 0 To a solution of B-3 (Scheme B where and are the same and equal to H, R,.
3 and RB, are the same and equal to CH 3 and stereochemistry is (17.46 g, 0.11 mol) in acetonitrile (250 mL) at ambient temperature was added di-t-butyl dicarbonate (25.64 g, 0.117 mol) followed by N.N-diisopropylethylamine (16.9 mL. 0.097 mol). The reaction mixture was stirred for 2 days and volatiles remove in vacuo. The residue was slurried in WO 99/67230 WO 9967230PCT/US99/1 4233 -92diethyl ether and filtered through a pad of celite. The filtrate was washed with 0. 1 N HC1, water, brine, dried (Na 2 filtered and concentrated in vacuo. Crystallization of the clear oil from hexane provided the title compound (3.85 g) as a white solid: mp 125- 126 0 C; (Lit. 1 14'C: Woodward, Heusler, Gosteli, Naegeli. Oppolzer, W.; Ramage, Ranganathan, Vorbruggen. H. J Am. Chem. Soc. 1966. 88, 852) H NMR (CDCI3) 868.70 (1 4.89 (1 3.27 (2 1.81 (6 MS (ESI-) for C, ,H,,N0 4 S mnlz 260.1 Preparation 17 and Example 72.
-I[
4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl ]-2-methoxy-2oxo-ethyllamino]carbonyl]-2.2-dimethyl-3thiaoidinecarboxlic acid 3-(1,1 dimethylethyl)ester (Scheme B, B-6: where RB-I and RB are the same and equal to H, R~ and RB~ r h same and equal to CH 3 Y is CO 2
R
3 is t-butyl, R,.
5 is OCH 3 R, is 4+[2,6dichlorobenzoyl)amino]phenyl and stereochemistry is HC// S H 0 0 0 -71\ 0 0CI To a cooled suspension of B-4 (Scheme B where and RB- 2 are the same and equal to H. and RBA are the same and equal to CH, Y is CO 2 R, is i-butyl and stereochemnistry is (1.0 g, 3.83 mmol) and HOBt (638 mg, 4.17 mmol) in CH 2 C1 2 mL) was added a solution of EDC (799 mg, 4.17 mnmol) in CH 2 C, (20 mL). After 30 min at 0-5'C, B-5 (Scheme B where RB-5 is OCH 3 R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemnistry is (1.93 g, 4.77 mmol) was added followed by 4methylmorpholine (520 piL, 4.77 mmol). The reaction mixture was gradually warmed to room temperature. stirred an additional 18 h and diluted with CH-,CI,. The organic layer was separated and washed with 0. 1 N HC1. sat, aqueous NaHCO3., brine, dried (Na-,S0 4 WO 99/67230 WO 9967230PCT/US99/I 4233 -93filtered and concentrated in vacuo. Flash chromatography of the residue using CH,C1 2 /acetone as eluant afforded the title compound (2.05 g) as a white solid: IR (mull) 1746, 1666. 1606, 1562, 1537, 1515, 1432, 1413, 1347. 1325, 1259, 1214, 1195, 1169, 799 'H NMR (CDC1 3 8 7.54 (2 7.3 2 (4 7.20 (2 6.90 (1 4.78 (2 3.74 (3 3.17 (4 1.74(3 1.43 (9 3 C NMR (CDC13) 8 171.6, 136.7, 136.2, 135.9, 132.7, 132.4, 132.3, 131.0, 130.6, 130.4, 130.0, 129.9, 128.2, 127.9, 120.5, 120.2, 120.1, 67.3, 53.4, 52.4, 42.8. 37.3, 36.9, 34.9. 30.9, 28.3; MIS (ESI+) for
C
2
,H
33 C1 2
N
3 0 6 S m/z 6 10 MIS (ESI-) for C ,H3Cl,N 3 0 m 07.9 Anal.
Calcd for C,H 3 3C1 2
N
3
O
6 S 0.13 H 2 0: C, 54.88; H. 5.47; N, 6.86. Found: C. 54.66; H, 5.5 7; N. 6.73. Water 0. 37.
Example 73.
,S I-Carboxy-2-[4-[(2,6dichlorobenzoy)aminopheny]ethyl] amino carbony]2,2dimethyl-3 thiazolidinecarboxylic acid 1, 1 -dimethylethyl)ester (Scheme B, B-7: where and RB.
2 are the same and equal to H, RB.
3 and R.4 are the same and equal to CH 3 Y is CO 2
R
3 is t-butyl, R, is 4-[2,6dichlorobenzoyl)amino]phenyl and stereochemnistry is H1CN1f-i N-AOH 0 o 0 c N Ci1 Example 73 was prepared from example 72 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1738, 1665, 1606. 1562, 1535, 1516, 1432, 1413, 1347, 1259, 1213. 1194. 1167, 799. 777 cm"; 'H NMR (CD3CN) 8 8.89 (1 7.54 (2 7.41 (3 7.25 (2 4.66 (2 3.15 (4 1.72 (3 1.70(3 1.35 (9 3 C NMR
(CD
3 CN) 8 177.3. 171.8, 163.4, 137.9, 137.1, 133.9. 132.7. 132.3. 131.3, 129.1, 120.7, 81.2. 79.3, 68.0. 54.5. 3 7.4. 28.4; MIS (ESI-) for CNO 6 mz 593.9 Anal.
WO 99/67230 PCT/US99/14233 -94- Calcd for C 27
,H
3 ,ClN 3 0 6 S 0.5 H,O: C, 53.56; H, 5.33; N, 6.94. Found: C, 53.77; H, 5.39; N, 6.70.
Example 74.
4 2 6 -Dichlorobenzoyl)amino]-N-[[(4R)-2,2-dimethyl-4-thiazolidinyl]carbonyl]-Lphenylalanine (Scheme B, B-8: where and R,,B 2 are the same and equal to H, R,.
3 and R,4 are the same and equal to CH 3 R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is
OH
0 ci
C
Example 74 was prepared from example 73 by the procedure described in preparation 4.
Physical data as follows: IR (mull) 3244, 3192, 3049, 1726, 1664, 1605, 1578, 1562, 1541, 1516, 1432, 1414, 1327, 1195. 799 'H NMR (DMSO-d 6 6 13.00 (1 10.69 (1 7.52 (6 7.24 (2 4.52 (2 3.12 (2 2.93 (1 1.62 (9 "C NMR 8 166.9, 136.8, 136.0. 133.5. 131.8, 130.9, 129.5, 129.4, 127.9. 120.4, 120.2, 120.1, 60.8, 54.3, 54.0, 36.2. 36.0. 25.1; MS (ESI+) for C 22
H
23
CN
3 0 4 S m/z 496.2 MS (ESI-) for C, 2
H,
3 C, N30 4 S m/z 494.2 Anal. Calcd for C,2H2, 3 C1,Nl2 3 0 4
S
HCI 0.50 H 2 0: C, 48.76; H. 4.65; N, 7.75. Found: C, 48.56; H, 4.72; N, 7.49.
Example 4 2 .6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxo-ethyl]amino]carbonyl]-2.2-dimethyl-3-thiazolidinecarboxylic acid 3-(1,1dimethylethyl)ester (Scheme B, B-6: where RB, and are the same and equal to H, R,3 and R.
4 are the same and equal to CH3, Y is CO,, R 3 is r-butyl, R, is dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 WO 9967230PCTIUS99/1 4233 0 Examle 5 ws pepaed a decried n SHem rmDcsen n ctn sn i -bty icront t fr tereusiecabaae.Pyscl aa s olos J mul bty m/z6079 M-Hnal. Cao c form the 3
C
2 3 0S Cuiit 55bt 08;sca Ht 5.45 Nlos 6.88 (l l).1 175256. oud:6, 1654.7, 547; N51, 6.78 43,14,135 29,11,126 (Schme6, B-7.4 where, RB0.9, and1299 1B 2 82105 are.4 th6sm7adeqa3t, 53.
3 and5 R.
4 re.th Sam a.5 nd:lt C, Yis7C0H .47 is 6-ut.78 s (26 stechmiry is 57)).thy-') thazliiecrbxli aid34 -imthlehO)ese (Scheme~~~~~ BB7 hr B1adR- r h aeadeult n r h WO 99/67230 PCT/US99/14233 -96- Example 76 was prepared from example 75 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3280, 1739, 1665, 1606, 1562. 1535, 1516, 1432, 1413, 1348, 1272, 1259. 1195. 1167, 799 'H NMR (CD3CN) 6 8.83 (1 7.55 (2 7.43 (3 7.22 (2 6.83 (1 4.68 (2 3.07 (5 1.73 (6 1.40 (9 H); "C NMR (CD 3 CN) 172.6. 163.3, 137.9. 133.4, 133.3, 132.7. 132.1, 131.1, 130.2, 130.1, 129.1, 128.1, 128.0, 120.6, 119.5, 54.1, 37.4, 29.4, 28.5, 27.6; MS (ESI+) for
C
27 Ha 3 C1 2
N
3 0 6 S m/z 595.9 MS (ESI-) for C 2 7
H
3 ,C1 2
N
3 0 6 S n/z 593.7 MS (FAB) m/z (rel. intensity) 596 16), 598 596 500 499 498 497 496 173 116 57 HRMS (FAB) calcd for
C,
7
H
3 ,C12N 3 0 6 S+H, 596.1389. found 596.1364; Anal. Calcd for C2,H 3 ,C1,N 3 0OS
H
2 0: C, 53.56; H, 5.33; N, 6.94. Found: C, 53.86; H, 5.35; N, 6.90.
Example 77.
2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl] -1-thia-4-azaspiro[4.4]nonane-4-carboxylic acid 4-ethyl ester (Scheme B, B-6: where RB., and RB.
2 are the same and equal to H, RB.
3 and RB. together form a carbocyclic ring of 5 atoms, Y is CO,, R 3 is ethyl, Rs is dichlorobenzoyl)amino]phenyl and stereochemistry is
O
N
O
Example 77 was prepared as described in Scheme B from D-cysteine and cyclopentanone using ethyl chloroformate to form the requisite carbamate. Physical data as follows: IR (mull) 1760, 1739, 1694, 1656. 1607, 1560, 1543, 1517, 1445. 1429, 1411, 1334, 1273, 1253, 1116 'H NMR (CDCI,) 5 7.56 (2 7.44 (1 7.34 (3 7.11 (2 6.70 (1 4.84 (2 4.14 (2 3.74 (3 3.12 (4 2.67 (1 2.51 (1 1.73 (6 H), 1.25 (3 "C NMR (CDCI 3 8 171.4, 170.7, 162.3. 136.3. 135.8, 132.5, 132.4, 131.0, 130.1, 128.2. 120.2, 66.4. 62.1. 53.1, 52.5, 37.3, 32.3. 31.9. 25.1. 24.6. 14.5, 14.1; MS WO 99/67230 WO 9967230PCT/US99/1 4233 -97- (ESI+) for C-, 8
H
31 C1 2
N
3 0 6 S m/z 630.0 HRMS (El) calcd for C.,,H 31 C1.IN 3
O
6
S
607.13 10, found 607.1315; Anal. Calcd for C, 8 3 1 CINOS 0.75 HAO C, 54.06; H, 5.27; N, 6.90. Found: C, 53.98; H, 5.16; N. 6.72.
Example 78.
I-Carboxy-2-[4-[(2,6dichlorobenzoy l)amino]phenyllethyl ]amino] carbonyl -thia-4-azaspiro nonane-4carboxylic acid 4--ethyl ester (Scheme B, B-7: where RB-1 and R,- 2 are the same and equal to H, RB.
3 and RB,4 together form a carbocyclic ring of 5 atoms, Y is CO., R 3 is ethyl, R, is dichlorobenzoyl)amino]phenyl and stereochemistry is
&PN'(N'OH
0 0' ci Example 78 was prepared from example 77 by the procedure described in preparation 6.
Physical data as follows: JR (mull) 3276, 1664, 1606, 1562, 1537, 1515, 1445, 1432, 1413. 1335, 1273, 1239, 1195, 1116, 799 cm-1; 'H NMR (CDC 3 8 8.08 (1 7.62 (2 H), 7.24(3 7.14 (2 6.72 (1 4.85 (2 4.13 (2 3.21 (4 2.53 (2 1.75 (6 1.22 (3 1 3 C NMR (CDC 3 8 175.2, 171.2, 162.6, 136.7, 135.7, 132.3, 132.0, 130.7, 130.2, 127.9, 120.2, 66.3, 62.2, 53.2, 37.3, 36.6, 32.3, 25.2, 24.6, 20.5, 14.5; MS (ESI+) for 9 C1,N 3 0 6 S m/z 593.8 MS (ESI-) for C2,H, 9 C1,N 3 0 6 S m/z 591.8 HRMS (FAB) calcd for C 27
H,
9 Cl,N 3 0 6 S+H, 594.1232, found 594.1226; Anal.
Calcd for C, 7
H
2 9 C1 2
N
3 0 6 S 0.37 H,O: C. 53.95; H, 4.99; N, 6.99. Found: C. 54.28; H, 10; N, 7.03. Water 1. Example 79.
[S4.R* 1 4 2 6 -Dichlorobenzoi)aminophenyllmethyl]2methoxy.2oxoethyl]amino]carbonylj-1 -thia-A--azaspiro[4 .5]decane-4-carboxylic acid 4-ethyl ester WO 99/67230 WO 9967230PCTIUS99/1 4233 -98- (Scheme B, B-6: where RB., and are the same and equal to H, RB-l and RB 4 together form a carbocyclic ring of 6 atoms,Y is COD, R 3 is ethyl, R, is 4-[2,6dichlorobenzoyl)amino]phenyl and stereochemistry is N HCi1 Example 79 was prepared as described in Scheme B from D-cysteine and cyclohexanone using ethyl chioroformate to form the requisite carbamnate. Physical data as follows: IR (mull) 1745, 1704, 1683, 1668, 1607, 1561, 1538, 1514, 1431, 1413, 1327, 1269, 1213, 1196, 1117 'H NMR (CDCl 3 6 7.57 (2 7.46 (1 7.33 (3 7.11 (2 6.72 (1 4.88 (2 4.14 (2 3.74 (3 3.13 (4 2.51 (1 1.69 (8 1.22 (5 H); 3 CNMR (CDCI 3 )5 171.4, 170.7, 168.1, 162.3, 136.3, 135.7, 132.4, 131.0, 128.2, 120.3, 66.9, 62.8, 62.0, 55.1, 53.1, 52.5, 42.0, 39.9, 37.3, 36.9. 31.1. 29.6, 27.6, 27.1, 26.0, 25.3, 24.7, 23.1, 14.5; MIS (ESI-) for C 29
H
33 C1 2
N
3 0 6 S m/z 621.9 HRMS (FAB) calcd for
C
29
H
33 C1 2
N
3 0 6 S+H, 622.1545. found 622.1536.
Example dichlorobenzoyl)amino]phenyl]ethyl~amino]carbonyl]1 -thia-4-azaspiro [4.5]decane-.4carboxylic acid 4-ethyl ester (Scheme B, B-7: where and R, 2 are the same and equal to H, RB.
3 and R, 4 together form a carbocyclic ring of 6 atoms, Y is CO,, R 3 is ethyl, R, is 4-(2,6dichlorobenzoyl)Amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -99- Example 80 was prepared from example 79 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3276, 1710, 1664, 1606, 1562. 1537, 1515, 1432, 1413. 1329, 1272, 1256, 1195, 1117. 800 cm-; 'HNMR (CDCI 3 8 8.07 (1 7.62 (2 H), 7.24 (3 7.14 (2 6.75 (1 4.88 (2 4.12 (3 3.14 (4 2.74 (1 2.50 (1 1.69 (6 1.19 (5 "C NMR (CDC1 3 6 175.4, 175.1. 171.3. 162.6, 136.7, 135.7, 132.3, 132.0, 130.7, 130.3. 130.1, 128.0, 120.2, 66.8, 62.1, 53.2. 42.0, 36.9, 30.5, 27.0, 26.0, 25.3, 25.0, 24.6, 20.5, 14.4, 3.7; MS (ESI+) for C 2 8
H
3 ,CIlN 3 0 6 S m/z 608.1 MS (ESI-) for C, 8
H
3 ,C1,N 3 0 6 S m/z 605.9 HRMS (El) calcd for
C,,H
31 CI2N 3 0S 607.1310, found 607.1309; Anal. Calcd for C 28 sH 3
,CN
3 0 6 S 0.3 H,O: C, 54.78; H, 5.19; N, 685. Found: C. 54.56; H. 5.24; N, 6.90. Water 0.87.
Example 81.
4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-2,2,5,5-tetramethyl-3-thiazolidinecarboxylic acid 3-ethyl ester (Scheme B, B-6: where RB.2, R,.3 and RB- are the same and equal to CH 3 Y is CO 2
R
3 is t-butyl, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is 0 oil^ 0 C Example 81 was prepared as described in Scheme B from D-penicillamine and acetone using ethyl chloroformate to form the requisite carbamate. Physical data as follows: IR (mull) 1748, 1666, 1606, 1562, 1538, 1516, 1432, 1413, 1327, 1275. 1233, 1215, 1195, 1080, 799 'H NMR (CDCI,) 8 7.55 (2 7.33 (4 7.09 (3 6.59 (1 4.91 (1 4.47 (1 4.16 (2 3.75 (3 3.13 (2 1.92 (3 1.75 (3 1.67 (3 H), 1.60 (3 1.21 (3 3 C NMR (CDCI 3 171.5, 170.0, 162.2, 136.1. 135.7. 132.5, 132.3. 131.9. 130.9, 130.7, 129.7. 128.1. 120.3, 120.1, 120.0, 61.6. 61.4. 52.9, 52.4, 52.2, WO 99/67230 WO 9967230PCTIUS99/14233 -100- 49.3, 37.4, 3 7.3. 34.1. 31.4, 30.8, 29.5, 24.9; MS (ESI-) for C,,H 33 C1IN 3 0 6 S m/z 607.9 (M- HIRMS (El) calcd for C 2
,H
33 Cl 2
N
3 0 6 S 609.1467, found 609.1461; Anal. Calcd for C,H3 3 C1 2
-N
3 0 6 S 0. 19 H,O: C, 54.77; H, 5.48; N. 6.84. Found: C, 55.00; H, 5.48-, N. 6.78.
Water 0.56.
Example 82.
I-Carboxy-2-[4-[(2,6dichlorobenzoylI)aminolpheny l]ethyl] amino] carbonyl ]2,2,5 ,S-tetramethyl1-3thiazolidinecarboxylic acid 3--ethyl ester (Scheme B. B-7: where RB-. RB.
2
RB-
3 and are the same and equal to CH 3 Y is CO, R3 is i-butyl, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is
H
N yN -A.OH 0 0 cI Example 82 was prepared from example 81 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3275, 1750, 1735. 1678, 1666, 1609, 1562, 1543, 1516, 1432. 1413, 1333, 1276, 1195, 1077 cm'; 'HNMR (DMSO-d 6 8 12.48 (1 10.63 (1 8.36 (1 7.53 (5 7.19 (2 4.49 (2 3.96 (2 3.03 (1 2.79 (1 H), 1.78 (6 1.48 (3 1. 15 (3 0.90 (3 3 C NMR (DMSO-d 6 6 172.9, 168.7, 161.7, 152.3, 137.0. 136.3, 132.9, 131.2, 131.1, 129.5, 128.1, 119.2, 74.7, 71.3, 60.5, 53.5, 49.0, 48.4, 38.3, 36.7,.33.5. 31.8, 28.1. 24.7, 21.0, 14.0; MIS (ESI+) for C, 7
H
31 C1 2
N
3 0 6 S m/z 595.8 MIS (ESI-) for C 27
H
3 1 C1 2
N
3 0 6 S m/z 593.8 FIRMS (FAB) calcd for
C.,
7
H
31 C1,-N 3 0 6 S±H, 596.1389, found 596.1362, Anal. Calcd for C 27
H
31 C1 2
N
3 0 6 S 0.56 H,O: C, 53.46, H, 5.34; N, 6.93. Found: C, 53-73, H, 5.35; N, 6.73. Water 1.67.
Example 83.
N-[[(4S)-3I-Acetyl-22-dimethyl-4-thiazolidin1 ]carbonyl]-4--[(2,6dichlorobenzoyl)amino]-L-phenvlalanine methyl ester WO 99/67230 WO 9967230PCTIUS99/1 4233 -101- (Scheme B, B-6: where RB., and RB- 2 are the same and equal to H. 3 and RB, are the same and equal to CH., Y is CO. R, is methyl, R, is 4-[(2,6-dichlorobenzoyl)arnino]phenyl and stereochemistry is HC 0 CI N~ 1 Example 83 was prepared as described in Scheme B from D-cysteine and acetone using acetyl chloride to form the requisite amide. Physical data as follows: IR (mull) 1745, 1682, 1662, 1628, 1610, 1579. 1561, 1541, 1515, 1431, 1412, 1326. 1270. 1240, 1211 cm- 1; 'H NMR (CDC 3 867.55 (3 7.32 (3 7.19 (1 7.11 (1 6.66 (1 4.89 (1 H), 4.60 (1 3.78(3 3.24 (4 2.04 (3 1.87 (3 1.79(3 3 C NMR (DMSOd 6 8 171.6, 171.5, 169.9, 169.4, 167.9, 161.8, 137.1, 136.3. 132.8. 132.6. 131.1, 129.5, 128.1, 119.2, 72.6, 66.5, 53.7, 53.3, 52.0, 51.9, 35.8, 31.7, 29.0. 26.9, 24.7, 24.5; MS (ESI±) for C 25
H
27 C1 2
N
3 0 5 S m/z 551.9 HRMS (El) calcd for C, 5
H
27 C1 2
N
3 0 5 551.1049, found 551.1053; MS (El) mlz (rel. intensity) 551 351 349 (68), 278 186 (14 175 173 158 116 100 99 (69).
Example 84.
N-[[(4S)-3-Acetyl-22-dimethyl-4-thiazolidinyl]carbonyl]-4-[(2,6dichlorobenzoyl)amino]-L-phenylalanine (Scheme B, B-7: where RB., and RB.
2 are the same and equal to H. RB 3 and R,,4 are the same and equal to CH 3 Y isCO0. R, is methyl, R, is 4-[(2,6-dichlorobenzoyl)aminojphenyl and stereochemnistry is WO 99/67230 PCT/US99/14233 -102- Example 84 was prepared from example 83 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3279, 1723, 1661, 1608, 1562, 1542, 1516, 1432, 1413. 1349. 1329, 1270, 1238, 1207, 1195 cm'; 'H NMR (DMSO-d 6 8 12.47 (1 10.62 (1 7.51 (5 7.18 (2 4.80 (1 4.67 (1 4.47 (1 2.98 (3 1.68 (9 H); 1 C NMR (DMSO-d 6 184.2, 172.6, 171.9, 169.8, 169.2, 167.8, 136.9, 136.3. 133.1, 131.2, 131.0, 129.5, 128.1, 119.2, 72.6, 66.6, 53.7. 53.3, 36.2, 31.7, 29.0, 27.0. 24.8, 24.6, 21.0; MS (ESI+) for C 2 4
H
25
CI
2
N
3 0sS m/z 538.0 MS (ESI-) for C 2 4 ,H2CIN 3 0S m/z 535.9 HRMS (FAB) calcd for C2 4
H
25 C1 2 NOS+H, 538.0970, found 538.0961.
Example dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-2,2-dimethyl-y-oxo-3thiazolidinebutanoic acid (Scheme B, B-7: where RB., and RB 2 are the same and equal to H, RB.
3 and RB 4 are the same and equal to CH 3 Y is CO, R 3 is CH,CH,CO 2 H, R, is dichlorobenzoyl)amino]phenyl and stereochemistry is 0 N 'OH HO 0 o. H CI Example 85 was prepared as described in Scheme B from D-cysteine and acetone using methyl succinyl chloride to form the requisite amide. Physical data as follows: IR (mull) 3264, 3125. 3071, 1724, 1658, 1607, 1562, 1537. 1517, 1432, 1414, 1326, 1241. 1195, 1181 'H NMR (DMSO-d,) 8 12.31 (1 10.65 (1 8.20(1 7.50 (5 7.19 (2 5.81 (1 4.50 (1 3.02 (3 2.30 (3 1.71 (6 "C NMR (DMSO-d 6 6 174.1. 172.8, 169.4, 161.9, 137.1, 136.5, 133.2, 131.4, 131.3, 129.7, 128.3, 119.3, 73.1, 65.7. 53.5. 36.3, 32.0, 30.5, 29.1, 29.0. 27.1: MS (ESI+) for C, 6
H
27 C12N 3 ,S m/z 595.9 MS (ESI-) for C,6H,,CINOS m/z 593.8 Anal. Calcd for C 6 ,,H7Cl 2
N
3 0,S WO 99/67230 WO 9967230PCTIUS99/14233 -103- 0.51 H,0: C. 51.55; H. 4.66;, N, 6.94. Found: C, 51.71: H. 4.85; N, 6.93. Water (KF): 1.53.
Example 86.
[S-(R*,R*)]-4-[[[1-[[4-[(2,6-Djchlorobenzoyl)amjno]phenvl]methylp2methoxy-2 oxoethyl]amino]carboriyl]-2,2,5 ,5-tetramethyl-y-oxo--3-thiazolidinebutanoic acid methyl ester (Scheme B, B-6: where RB-25 RB- 3 and R 8 are the same and equal to CH 3 Y is CO,
R
3 is CH 2
CH-,COCH
3 R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemnistry is 00 0 C0
HH
Example 86 was prepared as described in Scheme B from D-penicillamine and acetone using methyl succinyl chloride to form the requisite amide. Physical data as follows: IR (mull) 3287. 1741, 1660. 1608, 1562, 1540, 1516, 1432, 1413, 1323, 1267, 1241, 1225, 1196, 1168 'H NMR (CDCI 3 8 7.55 (2 7.29 (3 7.13 (2 6.94 (1 4.95 (1 4.39 (1 3.74 (3 3.67 (3 3.27 (1 3. 10 (1 2.80 (1 2.57 (3 H), 2.18 (1 1.95(3 1.77(3 1.72(3 1.68(3 H) 1.63(3 1 3 C NMR (CDCl 3 6 173.3, 171.6, 170.3, 169.9. 162.4, 136.3, 132.5, 132.4, 131.0. 130.2, 129.8, 128.2, 120.8, 120.7, 120.5, 73.8, 53.1, 52.6, 51.8, 49.9, 40.0, 37.4, 33.9. 31.6, 31.4, 29.4, 29.1, 24.3; MIS (ESI+) for C 30
H
35 C1 2
IN
3 01S m/z 652.1 Anal. Calcd for C 3 0
H
35 C11,N 3 0 7 S 0.31 C, 54.75; H, 5.45: N. 6.38. Found: C, 55.04; H, 5.50: N, 6.69. Water 0.84.
Example 87.
-Acetvl-2.2.5 ,5-tetramethyl-4-thiazolidinyl]carbonyl]-4-[(2,6dichlorobenzoyI)aminoj-L-phenylalanine methyl ester WO 99/67230 WO 9967230PCTIUS99/1 4233 -104- (Scheme-B. B-6: where RB.1, RB-2, RB.
3 and are the same and equal to CH,, Y is CO, R3, is methyl, R, is 4-[(2,6-dichlorobenzoyl)aminolphenyl and stereochemnistry is
H
H
3 CO 0 0 C1
NX
Example 87 was prepared as described in Scheme B from D-penicillamine and acetone using acetyl chloride to form the requisite amide. Physical data as follows: IR (mull) 1741. 1692. 1676, 1629, 1606, 1563, 1538, 1516, 1431, 1410, 1352, 1320, 1276, 1251, 1223 'H NMR (DMSO-d 6 5 10.70 (1 8.57 (1 7.56 (4 7.22 (2 4.63 (1 4.49 (1 3.65 (3 3.30(1 3.12 (1 2.84 (1 1.92 (3 1.82 (3 H), 1.78(3 1.51 (3 0.80 (3 3 C NMR (DMSO-d 6 8 184.2, 171.9. 168.9, 168.1, 161.8, 137.1, 136.3, 132.4, 131.3, 131.2, 131., 129.4, 128.1, 119.3, 75.9, 72.8, 53.5, 52.0, 49.2, 48.4, 36.0, 33.5, 31.9, 27.6, 24.9, 24.5; MIS (ESI+) for C 27
H
3 C 2-N 3 0 S /z 580.1 MIS (ESI-) for C,,H 31 C1 1
N
3 0 5 S m/lz 577.9 Anal. Calcd for C 27
H
31 Cl 2
N
3 0 5
S
0.12 H,O: C, 55.66; H, 5.40; N, 7.21. Found: C, 55.68; H, 5.39; N. 7.16. Water (KF): 0.36.
Example 88.
I-Carboxy-2--[4-[(2,6dichlorobenzoyl)amnino]pheny] ethyl] amino]carbonyl]-2,2,5,5-tetramethyl-xo-3.
thiazolidinepentanoic acid (Scheme B. B-6: where RB-1, RB.
2
RB-
3 and RB,. are the same and equal to CH3, Y is GO, R, is CH,CHCHCo 2 H, R, is 4 -[(2.6-dichlorobenzoyl)amino]phenyl and stereochemnistry is WO 99/67230 WO 9967230PCTIUS99/1 4233 -105- 0 C HO- 1
N
0
HI
C1 Example 88 was prepared as described in Scheme B from D-penicillamine and acetone using methyl glutaryl chloride to form the requisite amide. Physical data as follows: 'H NMR (DMSO-d 6 6 10.65 (1 8.45 (1 7.53 (5 7.22 (2 4.51 (1 3.11 (3 2.77 (1 2.21 (4 1.89 (3 1.82(3 1. 11 (2 1.48(3 0.77(3 H); 3 CNMR (DMSO-d 6 8 184.2, 174.2, 173.1, 170.1, 168.7, 161.7, 137.0, 136.3. 133. 1.
131.3, 131.1, 129.4, 128.1, 119.2, 74.9, 73.0, 53.8, 49.3, 36.1, 34.8, 33.5, 32.7. 31.9, 27.7, 24.5, 20.0; MIS (ESJ+) for C 29
H
33 C1 2
N
3 0 7 S m/z 638.0 MIS (ESI-) for
C
29
H
33 C1 2
N
3 0 7 S m/z 635.9 HRMS (FAB) calcd for C 29
H
33
C,N
3 0 7 5+H, 638.1494, found 638.148 1; Anal. Calcd for C 29
H
3 3 C1,N 3 0,S 0.75 H 2 0: C, 53.42; H, 5.33 N, 6.44.
Found: C, 53.20; H, 5.26; N, 6.45.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -106- Scheme C NH-Fmoc 0 H N IIojo~ -NH-Fmoc C-1 01 Rink-amide-MBHA resin Q NH 2 C-2 I 0 Fmoc-HN 0 Q-N NH-Fmoc
HC-
R
0 Q-N NH 2 H
R
RC-1 Fmoc 0 oRC
RKS
I N N' RC4
R
5 0 FmOC oRC
RC.
0H S RC Q N XRC- C-8
R
WO 99/67230 PCT/US99/14233 -107- Scheme C (continued) Rc S Rc-2 C-
R
QN N N RC C-8 -H J o H
R
1 RC2 RC 0 Rt Rc.- N N NRC4 C-9
R
H 0
Y-R
3
R
5 RC-, Rc.l RC.4 O"s
NH
2 R3 .y Rs Where: Rc.
1 Rc.2, and Rc-3 are defined independently as Rc 4 is defined as R 2 Scheme C describes a method for the preparation of examples of the formula Commercially available Rink Amide MBHA resin is deprotected under standard solidphase peptide synthesis conditions (Atherton. Sheppard R.C. Solid Phase Peptide Synthesis: A Practical Approach; IRL Press at Oxford University Press: Oxford, 1989) to afford the amine of formula C-2. Acylation with a commercially available or readily prepared amino acid residue of general C-3 affords the resin bound derivative of formula C-4. Removal of the Fmoc group under standard conditions provides amine of general structure C-5 which is acylated with a commercially available or readily prepared thiazolidine-4-carboxylic acid of general formula C-6 to afford the resin bound intermediate C-7. Standard Fmoc deprotection affords the resin bound amine of general formula C-8 which may be reacted with a variety of electrophilic reagents as described in Scheme A to afford resin bound amides, ureas. sulfonamides and carbamates of general structure C-9. Preparation 18 details an example of the reaction of a mixed carbonate to WO 99/67230 PCTIUS99/1 4233 -108afford a carbamate of general structure C-9 (where Y is equal to CO 2 Standard acidolysis affords the amide of general structure Preparation 18 and Example 89.
4 -[(2,6-Dichlorophenyl)methoxy]phenyl]methyl]-2-amino-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3 2 -(4-morpholinyl)ethyl] ester (Scheme C, C-10: where Rc.2, Rc.3 and Rc are the same and equal to proton, R 3 is 2- (4-morpholinyl)ethyl,
R
5 is 4-[(2,6-dichlorophenyl)methoxy]phenyl, Y is CO 2 and stereochemistry is s- O 0 N-Y NH2 0 ci0 To a mixture of Rink Amide MBHA resin (Scheme C, C-1) (Nova Biochem., 1.2 g, ca. 0.59 mmol) in methylene chloride (20 mL) was added a solution of piperidine in DMF 20 mL). A slow stream of nitrogen was bubbled through the mixture to effect mixing for 20 min. The resin was filtered and washed with DMF. The resin was suspended in a solution of piperidine in DMF 40 ml) and mixed for 40 min. The resin was filtered and washed with DMF, methylene chloride, methanol and methylene chloride to afford resin C-2, which was diluted with DMF (40 mL). To this mixture was added Fmoc-Tyr(2,6-Cl,-Bn) (Scheme C, C-3: where R 5 is dichlorophenyl)methoxy]phenyl and stereochemistry is (Advanced Chemtech, 1.32 g, 2.35 mmol), HOBt (0.36 g, 2.35 mmol), PyBOP (1.20 g, 2.35) and DIEA (1.03 mL, 5.90 mmol). The reaction was mixed for 4 h and the resin filtered and washed with DMF, methylene chloride, MeOH and methylene chloride to afford the intermediate resin-bound WO 99/67230 PCT/US99/14233 -109amino acid derivative C-4 (Scheme C, where R 5 is 4 2 ,6-dichlorophenyl)methoxy]phenyl and stereochemistry is which was used without characterization.
To a mixture of resin C-4 described above in methylene chloride (20 mL) was added a solution of piperidine in DMF 20 mL). A slow stream of nitrogen was bubbled through the mixture to effect mixing for 20 min. The resin was filtered and washed with DMF. The resin was suspended in a solution of piperidine in DMF ml) and mixed for 40 min. The resin was filtered and washed with DMF, methylene chloride, methanol and methylene chloride to afford the resin of structure C-5 (Scheme C, where R, is 4 2 6 -dichlorophenyl)methoxy]phenyl and stereochemistry is which was diluted with DMF (40 mL). To this mixture was added Fmoc-D-thiazolidine-4carboxylic acid (Scheme C, C-6: where Rc.E, R.
2 Re.3 and Rc, are the same and equal to proton and stereochemistry is (Advanced Chemtech, 832 mg, 2.35 mmol), HOBt (0.36 g, 2.35 mmol), PyBOP (1.20 g, 2.35) and DIEA (1.03 mL, 5.90 mmol). The reaction was mixed for 4 h and the resin filtered and washed with DMF, methylene chloride, MeOH and methylene chloride to afford the intermediate resin-bound derivative C-7 (Scheme C, where Rc., Re.
2 Rc.
3 and Rc are the same and equal to proton, R 5 is dichlorophenyl)methoxy]phenyl and stereochemistry is which was used without characterization.
To a mixture of resin C-7 described above in methylene chloride (20 mL) was added a solution of piperidine in DMF 20 mL). A slow stream of nitrogen was bubbled through the mixture to effect mixing for 20 min. The resin was filtered and washed with DMF. The resin was suspended in a solution of piperidine in DMF ml) and mixed for 40 min. The resin was filtered and washed with DMF, methylene chloride, methanol and methylene chloride to provide the intermediate of structure C-8 (Scheme C, where Rc,, Rc.2, Rc.
3 and Rc are the same and equal to proton, R 5 is dichlorophenyl)methoxy]phenyl and stereochemistry is which was diluted with methylene chloride (10 mL). To this mixture was added a solution of the mixed carbonate prepared from 4-(2-hydroxyethyl)morpholine (2.14 mL, 17.7 mmol) and N,Ndisuccinimidyl carbonate (4.53 g, 17.7 mmol) as described in preparation 8 in methylene chloride (20 mL) followed by triethylamine (0.33 mL, 2.36 mmol). The reaction was WO 99/67230 PCT/US99/14233 -110mixed for 24 h and the resin filtered and washed extensively with DMF, methylene chloride, MeOH, and anhydrous ethyl ether. The resin was dried in vacuo to afford the resin-bound carbamate C-9 (Scheme C, where Rc, R.
3 and Rc, are the same and equal to proton, R 3 is 2-(4-morpholinyl)ethyl, R, is dichlorophenyl)methoxy]phenyl, Y is CO, and stereochemistry is Resin C-9 was swelled with a minimum of methylene chloride (ca. 2 mL) and suspended with 95 aqueous TFA (20 mL). The mixture was mixed by magnetic stirring for Ih and filtered. The resin was washed with additional TFA (2 X 5 mL), followed by methylene chloride and methanol. The combined filtrates were evaporated in vacuo, and partioned between ethyl acetate and saturated aqueous sodium bicarbonate. The organic layer was separated and washed with brine, dried (MgSO4), filtered and evaporated in vacuo. The residue was purified by flash chromatography using methylene chloride/methanol (1 to as eluant to afford the title compound (215 mg) as an amorphous powder: IR (mull) 3288, 1676, 1657,1611, 1564, 1511, 1439, 1424, 1346, 1302, 1237, 1179, 1116, 1021, 767 'H NMR (CDCl 3 300 MHz) 8 7.24 (5 6.93 (2 5.20 (2 4.63 (3 4.34 (1 4.22 (2 3.63 (4 3.11 (4 2.50 (6 "C NMR (CDCLI, 75 MHz) 8 173.4, 170.1, 157.9, 154.5, 136.9, 132.0, 130.4, 129.1, 128.4, 115.1, 66.8, 65.2, 63.4, 57.1, 54.0, 53.7, 53.5, 49.4. 35.2; MS (EI) m/z (rel. intensity) 610 323 321 161 159 114(31), 113 100(99), 88 56 MS (FAB) m/z (rel. intensity) 611 (MH, 71), 614 613 612 (27), 611 123 114 113 112 107 100 HRMS (FAB) calcd for C 27
H
32 C1 2
N
4 0 6 S 611.1498, found 611.1494. Anal. Calcd for C 27
H
32 Cl 2
N
4 06S: C, 53.03; H, 5.27; N, 9.16. Found: C, 52.74; H, 5.17; N, 9.01.
Example 4 2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-amino-2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester (Scheme C, C-10: where Rc2, Rc-3 and Rc4 are the same and equal to proton, R 3 is ethyl, R 5 is 4-[(2,6-dichlorophenyl)methoxy]phenyl, Y is CO, and stereochemistry is (S, WO 99/67230 PCT/US99/14233 -111o VYNH2 N- N "AN CH l 0
C,
Example 90 was prepared as described in Scheme C. Physical data as follows: IR (mull) 3369, 3308, 3192, 1713, 1667, 1650, 1629, 1539, 1513, 1441, 1344, 1290, 1240, 1016, 768 cm'; 'H NMR (CDCI 3 300 MHz) 5 7.33 (3 7.16 (2 6.95 (2 5.25 (2 H), 4.48 (4 4.20 (2 3.03 (4 1.26 (3 3 C NMR (CDC 3 75 MHz) 6 173.6, 170.5, 157.8, 154.7, 136.8, 313.9, 130.4, 130.2, 129.0, 128.4, 114.9, 67.9. 65.1. 63.1, 62.5, 53.8, 36.5, 14.3; MS (EI) m/z (rel. intensity) 525 323 322 321 267 265 163 161 160 159 88 MS (FAB) m/z (rel. intensity) 526 58), 528 527 526 321 188 161 160 159 107 88 HRMS (FAB) calcd for C 23
H
25 C1 2
N
3 OsS 526.0970, found 526.0942. Anal. Calcd for C 23 H,25C1N 3 0,S: C, 52.47; H, 4.79; N, 7.98. Found: C, 52.34; H, 4.81; N, 7.90.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -112- Scheme D
R
Fmoc, N OH H 0 1 H 2 NOt-Bu D-2
R
HH
H
2 o, N 0OL -3 0
RD-,
R3S_ R 0 2 0 IFMOC 0 R RD-3~ D-6 R4S D-1 0 RD1 0
R
RD-3 N,
R.SR
0 0 ND~
N
D< R H, -8
RD-
1RD- Rk' NO 09 IR S R 0 H 09 WO 99/67230 PCT/US99/14233 -113- RD-2, and R- 3 are defined independently as RD4 is defined as R,.
Scheme D describes a method for the preparation of examples of general formula D-9. Commercially available or readily prepared N-cc-Fmoc protected amino acids of general structure D-1 are coupled with O-(tert-butyl)hydroxylamine under standard coupling conditions as previously referenced to afford the t-butyl hydroxamate of general structure D-3. Standard Fmoc deprotection affords the intermediate amine of formula D-4.
Coupling of this amine with a commercially available or readily prepared N-a-Fmocthiazolidine-4-carboxylic acid of general structure D-5 affords the pseudodipeptide intermediate of general structure D-6. Standard Fmoc deprotection affords the intermediate amine of general structure D-7 which may be reacted under the variety of conditions described in Scheme A to afford amides, carbamates, sulfonamides and ureas of general structure D-8. Preparation 22 provides a specific example of the reaction of an amine of general structure D-7 with a carbonate to afford a carbamate of general structure D-8. Mild acidolysis affords the hydroxamate of general structure D-9.
Preparation 19.
(Scheme D, D-3: where R, is 4 -[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is H 0 Fmoc' N 0 cl To a cooled (0-5 solution of Fmoc-Tyr(2,6-C,-Bn) (Scheme D, D-l: where R 5 is 4- 2 ,6-dichlorophenyl)methoxy]phenyl and stereochemistry is (Advanced Chemtech, g, 10.7 mmol), HOBt (1.63 g, 10.7 mmol), O-(tert-butyl)hydroxylamine hydrochloride (Scheme D, D-2) (1.61 g, 12.80 mmol) in methylene chloride (30 mL) was added PyBOP (6.66 g, 12.80 mmol) followed by DIEA (6.51 mL, 37.35 mmol). The mixture was stirred at 0-5 OC for 1 hour, gradually allowed to warm to room temperature and stirred an additional 2 h. The mixture was diluted with methylene chloride and 0.25 N HCI, the organic layer separated and washed with saturated aqueous NaHCO 3 and brine, dried WO 99/67230 PCTIUS99/14233 -114- (MgSO 4 filtered and evaporated in vacuo. The residue was purified by flash chromatography using methylene chloride/methanol as eluant to afford the title compound (5.87 g) as an amorphous powder: 'H NMR (CDCI 3 300 MHz) 6 8.08 (1 H), 7.76 (2 7.54 (2 7.29 (9 6.95 (2 5.43 (1 5.30 (2 4.28 (4 3.06 (2 1.19 (9 3 C NMR (CDCI 3 75 MHz) 6 169.7, 157.8, 156.3, 143.6. 141.2, 136.8, 132.0, 130.4, 128.9, 128.4, 127.7, 127.0, 125.0, 119.9, 115.0, 82.3, 67.1, 65.1, 54.0, 46.9, 37.6, 26.0; MS (ESI+) for C3,H 34 CIlN2,0 m/z 632.9 MS (ESI+) for
C
3 5
H,
4 CI,N,0 5 m/z 654.9 Preparation (Scheme D. D-6: where RD., R.
2
RD.
3 and RD4 are the same and equal to proton. R, is 4- 2 6 -dichlorophenyl)methoxy]phenyl and stereochemistry is O H CN
NO
Fmoc c To a solution of D-3 (Scheme D, where R, is dichlorophenyl)methoxy]phenyl and stereochemistry is (5.87 g, 9.28 mmol) in anhydrous DMF (94 mL) was added diethylamine (9.40 mL. 90.84 mmol) at ambient temperature. The solution was stirred for 90 min and volatiles were removed in vacuo to afford the intermediate amine D-4 (Scheme D, where R, is dichlorophenyl)methoxy]phenyl and stereochemistry is as an oil which was used without further purification.
To a cooled (0-5 solution of Fmoc-D-thiazolidine-4-carboxylic acid (Scheme D, D-5: where RD., RD2, RD 3 and RD4 are the same and equal to proton and stereochemistry is (Advanced Chemtech. 3.93 g, 11.10 mmol) and HOAt (1.51 g, 11.10 mmol) in methylene chloride/DMF 30 mL) was added EDC (2.12 g, 11.10 mmol). The reaction mixture was allowed to stir for 15 min and a solution of the amine (D-4 described above) in methylene chloride/DMF 30 mL) was added followed by WO 99/67230 PCT/US99/14233 -115- DIEA (1.61 mL, 9.28 mmol). After 1 h at 0-5 OC, an additional equivalent of DIEA (1.61 mL, 9.28 mmol) was added and the mixture allowed to warm to room temperature. After stirring overnight, volatiles were removed in vacuo and the residue partitioned between ethyl acetate and 0.25 N aqueous HCI. The organic layer was separated and washed with water, saturated aqueous NaHCO 3 brine, dried (Na,SO 4 filtered and concentrated in vacuo. The residue was purified by flash chromatography using CH 2
C
2 /acetone containing isopropanol as eluant to afford the title compound (2.4 g) as an amorphous solid: 'H NMR (300 MHz, CDC13) 8 7.77 (2 7.55 (2 7.32 (7 7.12 (2 6.92 (2 6.70 (1 5.19 (3 4.55 (5 4.26 (2 3.30 (1 3.11 (3 1.14 (9 "C NMR (75 MHz, CDC1 3 170.0, 168.8, 158.0, 143.4. 141.3, 137.0, 132.1, 130.4, 128.5, 127.9. 127.2, 124.9, 120.1, 115.3, 82.5, 68.4, 65.3, 52.8, 47.1, 36.5, 26.2.
MS (ESI+) for C3,H 3 9
CI,N
3 0 6 S m/z 747.9 MS (ESI+) for C 39
H
3 ,C1,N 3 06S m/z 769.8 MS (ESI-) for C 3
,H
39 C1 2
N
3 0 6 S m/z 745.7 Preparation 21.
(Scheme D, D-7: where RD-2, RD.
3 and R, are the same and equal to proton, R, is 4- [(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is o 0I To a solution of D-6 (Scheme D, where RD.
2
RD.
3 and RD4 are the same and equal to proton, Rs is 4-[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is (500 mg, 0.67 mmol) in anhydrous DMF (7 mL) was added diethylamine (0.70 mL, 6.55 mmol) at ambient temperature. The solution was stirred for 90 min and volatiles were removed in vacuo. The residue was washed with ethyl ether/hexane to afford the title compound (352 mg) as an amorphous solid which was used without further purification: 'H NMR (300 MHz, DMSO-d6) 6 10.61 (1 8.27 (1 7.54 (2 7.44 (1 7.15 (2 6.94 (2 5.15 (2 4.50 (1 4.03 (2 3.75 (1 3.19 (1 2.82 (3 2.57 (1 H), WO 99/67230 PCT/US99/14233 -116- 1.06 (9 MS (ESI+) for C 24 HCl 2
N
3 0 4 S m/z 526.1 MS (ESI-) for
C
2 4
H
29 C1,N 3 0 4 S m/z 524.1 Preparation 22.
(Scheme D, D-8: where RD-2, RD 3 and RD4 are the same and equal to proton, R 3 is 2- (4-morpholinyl)ethyl, R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl, Y is CO 2 and stereochemistry is
O
0 N NX ,O o C a To a solution of 4-(2-hydroxyethyl)morpholine (1.22 mL, 10.05 mmol) in CH 3
CN
mL) at ambient temperature was added N,N-disuccinimidyl carbonate (2.49 g, 10.05 mmol) and triethylamine (4.20 mL, 30.15 mmol). The solution was stirred at room temperature for 4 h and concentrated in vacuo to give a viscous oil. The oil was dissolved in a minimal amount of methylene chloride (15 mL) and added to a solution of D-7 (Scheme D, where RD 2 RD-3 and R.
4 are the same and equal to proton, R, is dichlorophenyl)methoxy]phenyl and stereochemistry is S) (350 mg, 0.67 mmol), triethylamine (0.10 m, 0.74 mmol) and DMAP (1 mg) in CH 2 C12 (4 mL). The reaction mixture was stirred overnight and diluted with CHC1 2 (15 mL). Propylamine (8.6 mL, 100.5 mmol) was slowly added (exothermic) and the solution stirred vigorously for min, then diluted with water. The organic layer was separated and washed with 0.1 M HCI, saturated aqueous NaHCO 3 and brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using ethyl acetate/acetone as eluant afforded the title compound (251 mg) as an white powder: IR (mull) 3264, 1709, 1661, 1564, 1531, 1512, 1439, 1419, 1345, 1301, 1241, 1181, 1118, 1016, 767 cm'; 'H NMR (300 MHz, CDC 3 8 8.48 (1 7.36 (2 7.24 (1 7.17 (2 6.96 (2H), 5.23 (2 4.60 (3 4.31 (3 3.71 (4 3.33 (4 2.59 (6 1.15 (9 3
C
NMR (75 MHz, CDCl 3 168.9, 158.0, 137.0, 132.1, 130.5, 128.7, 128.5, 115.3, 82.5, WO 99/67230 PCT/US99/14233 -117- 66.8, 65.3, 63.4, 57.2, 53.7, 52.9, 36.4, 30.6, 29.3, 26.2, 19.1, 13.7; MS (ESI+) for
C
31
H
4 0 C1 2
N
4 0,S m/z 682.9 MS (ESI+) for C3,H 40 oC 2
N
4 0 7 S m/z 705.0 Anal. Calcd for C, 3
H
40 C1 2
N
4 0,S 0.35 H 2 0: C, 53.97; H, 5.95; N, 7.95. Found: C, 54.22; H, 6.11; N, 7.95. Water 0.91.
Preparation 23 and Example 91.
(Scheme D, D-9: where RD2, RD3 and RD 4 are the same and equal to proton, R 3 is 2- (4-morpholinyl)ethyl, R 5 is 4-[(2,6-dichlorophenyl)methoxy]phenyl, Y is CO 2 and stereochemistry is oS H ci Hydroxamate D-8 (Scheme D, where RD- 2 RD3 and RD. are the same and equal to proton, R 3 is 2-(4-morpholinyl)ethyl, R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl, Y is
CO
2 and stereochemistry is (150 mg, 0.22 mmol) was dissolved in anhydrous TFA (12 mL) at ambient temperature and gradually warmed to 40 OC. After 5 h at 40 OC, volatiles were removed in vacuo and the residue partitioned between ethyl acetate and saturated aqueous NaHCO 3 The organic layer was separated and washed with brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using methylene chloride/methanol as eluant afforded the title compound (51 mg) as an amorphous solid: IR (mull) 3273, 3229, 1708, 1652, 1564, 1546, 1511, 1439, 1422, 1346, 1236, 1180, 1114, 1022, 768 cm 'H NMR (300 MHz, DMSOd 6 8 10.70 (1 8.93 (1 8.32 (1 7.54 (2 7.45 (1 7.12 (2 6.93 (2 H), 5.16 (2 4.56 (2 4.36 (1 4.25 (1 4.06 (2 3.51 (4 3.15 (1 2.76 (3 2.36 (4 3 C NMR (75 MHz, DMSO-d 6 8 169.8, 167.9, 157.6, 136.5, 132.2, 132.0, 130.8, 130.5, 129.2, 114.7, 66.7, 65.3, 63.3, 59.1, 57.1, 53.8, 52.1, 37.8, 31.3, 30.1; MS (ESI+) for C 27
H
32 C1 2
N
4 0 7 S m/z 627.0 MS (ESI-) for C 27
H
32 Cl2N 4 0 7 S m/z 624.9 WO 99/67230 PCT/US99/14233 -118- Anal. Calcd for C 2 7
H
32 C1 2
N
4 0S 0.46 H 2 0: C, 51.00; H, 5.22; N, 8.81. Found: C, 51.34; H, 5.23; N, 8.67. Water 1.31.
Example 92.
(Scheme D, D-9: where RD,, RD 2 RD-3 and RD. are the same and equal to proton, R 3 is ethyl, R, is 4 2 6 -dichlorophenyl)methoxy]phenyl, Y is CO, and stereochemistry is (S,
S).
O
(N>YN .A NOH ~o H c 0 00 N.Y Ci Example 92 was prepared as described in Scheme D from Fmoc-Tyr(2,6-Cl,-Bn) using ethyl chloroformate to provide the requisite carbamate. Physical properties as follows: IR (mull) 3278, 1654, 1612, 1585, 1564, 1547, 1511, 1439, 1347, 1237, 1195, 1179, 1022, 782, 769 'H NMR (300 MHz, CDC1 3 8 7.39 (3 7.17 (2 6.94 (2 5.24 (2 4.53 (4 4.18 (2 3.15 (2 2.87 (2 1.28 (3 1 3 C NMR (75 MHz, CD30D)8 172.7, 170.1, 159.4, 156.2, 138.1, 133.7, 132.1, 131.6, 130.8, 129.8, 115.9, 66.3, 64.4, 63.6, 38.6, 36.8, 35.1, 15.0; HRMS (FAB) calcd for C 2 3
H
25 CL2N 3 0 6 S +H, 542.0919, found 542.0921; Anal. Calcd for C 23
H
25 C1 2
N
3 0 6 S: C, 50.93; H, 4.64; N, 7.75.
Found: C, 50.79; H, 4.79; N, 7.52.
Example 93.
(Scheme D, D-9: where RD., RD.
3 and RD 4 are the same and equal to proton, R 3 is 2- (1-piperidinyl)ethyl, R, is 4 2 6 -dichlorophenyl)methoxy]phenyl, Y is CO2 and stereochemistry is WO 99/67230 PCT/US99/14233 -119- Example 93 was prepared as described in Scheme D from Fmoc-Tyr(2,6-Cl2-Bn) using 1- (2-hydroxyethyl)piperidine to provide the requisite carbamate. Physical properties as follows: IR (mull) 3276, 1707, 1653, 1611, 1584, 1564, 1511, 1439, 1237, 1195, 1179, 1144, 1113, 1093, 1021 cm 'H NMR (300 MHz, DMSO-d 6 8 7.54 (2 7.44 (1 H), 7.12 (2 6.93 (2 5.16 (2 4.58 (2 4.36 (1 4.25 (1 4.00 (3 2.76 (3 2.31 (3 1.62 (1 1.42 (3 1.26 (8 3 C NMR (75 MHz, DMSO-d 6 8 172.2, 168.9, 167.3, 157.0, 135.9, 131.7, 131.4, 130.2, 129.9, 128.7, 114.1, 64.7, 63.0, 54.0, 51.6, 36.2, 30.9, 25.5, 24.7, 23.8, 22.0, 21.5, 13.9; MS (ESI+) for C 2 8
H
34
CI
2
N
4 0 6 S m/z 624.9 WO 99/67230 WO 9967230PCTIUS99/1 4233 -120- Scheme E N S E-1 0 0 -oN 0 E-2 0 0 N0 E- O2 1 E- 0 N O o E-6 WO 99/67230 WO 9967230PCTIUJS99/1 4233 -121- Scheme E (continued) 0
R
N
U N H 0 0 N
OH
O N H Preparation 24.
(Scheme E, E-2 where stereochemnistry is (2-Methoxy-2-oxoethylidene)-1 (phenylmethyl)-L-Proline 1,1-dimethylethyl ester 0 0 To a stirring solution of E-1 (Scheme E where stereochemistry is (3.62g, 12.4mmol), prepared by the method of Rapoport Am. Chem. Soc. 1984, 106. 4539), in WO 99/67230 PCT/US99/14233 -122- CH3CN (10mL) was added methyl bromoacetate (1.4mL, 14.9mmol). After stirring for CH2C12 (70mL) was added. The solution stirred for 10 minutes before Ph3P (4.89g, 18.6mmol) was added, and after 2 minutes Et3N (5.2mL, 37.3mmol) was added.
After stirring for 20h, the solution was washed with 1M NaH2PO4 (100mL), and the aqueous phase was extracted with CH2CI2 (50mL). The combined organic phases were washed with brine, dried (Na2SO4), filtered, and evaporated in vacuo. The resulting yellow oil/white solid was dissolved in CHC13 and chromatographed on silica gel (300g, 230-400 mesh, 70mm OD column, packed CHC13, eluted with CHC13, 3L, then 10:90 EtOAc-CHCl3, 250mL fractions) using the flash technique. Fractions 19-23 provided the title compound (3.23g) as a pale yellow oil. 1 H-NMR: (300MHz, CDC13): 8 7.17-7.36 4.75 (1 4.54 (1 4.20 (1 3.96 (1 3.61 (3 3.36-3.47 (1 3.08 (1 2.04-2.28 (2 1.41 (9 EI/MS (70eV) m/z (rel. intensity): 331 (M 17.3), 275 230 170 91 (base); IR (neat): 2979, 2948, 1735, 1692, 1600, 1454, 1435, 1414, 1369, 1299, 1277, 1184, 1137, 1059, 964, 843, and 789cm-1; HRMS: Calcd. for C19H25N10 4 331.1783. Found: 331.1771; [a]D 2 5 +1070 0.939, CH2C12).
Preparation (Scheme E, E-3 where stereochemistry is (1,1-dimethylethoxy)carbonyl]-l- (phenylmethyl)-2-pyrrolidineacetic acid methyl ester 0 Raney-Nickel (20g of a 50% slurry in H20) was washed with abs. EtOH (3x25mL) and suspended in abs. EtOH (50mL), and a solution of E-2 (Scheme E, where WO 99/67230 PCT/US99/14233 -123stereochemistry is (9.34g, 28.2mmol) in abs. EtOH (50mL) was added. After stirring for 3h, the Ra-Ni was removed by filtration, and the filtrate was evaporated in vacuo. The residue was dissolved in EtOAc (100mL), 5% Pt/C (3.0g) was added, and the mixture was hydrogenated under 50 psi H2 for 12 h. The catalyst was removed by filtration and the filtrate was evaporated in vacuo. The residue was chromatographed on silica gel (300g, 230-400 mesh, 70mm OD column, packed and eluted with 15:85 EtOAc-hexanes, 270mL fractions) using the flash technique. Fractions 5-8 provided the title compound (6.55g) as a clear, colorless oil. 1 H-NMR: (300MHz, CDC13): 6 7.22-7.34 (5 3.86 (1 3.79 (1 3.62 (3 3.21-3.29 (2 2.57 (1 2.33 (1 H), 1.64-2.05 (4 1.37 (9 EI/MS (70eV) m/z (rel. intensity):232 (base), 91 IR (nujol): 2977, 1739, 1454, 1437, 1367, 1295, 1251, 1197, 1153, 1074, 844, 753, and 699cm- 1 Anal: Calcd. for C19H27N 1 0 4 C, 68.44; H, 8.16; N, 4.20. Found: C, 68.39; H, 8.15; N, 4.11; [a]D 2 5 -22° (c 1.051, CH2CI2).
Preparation 26.
(Scheme E, E-4 where stereochemistry is (1,1-dimethylethoxy)carbonyll-l- (phenylmethyl)-2-pyrrolidineacetic acid
O
o-o To a stirring solution of E-3 (Scheme E, where stereochemistry is (2R,5S)) (2.00g, 6.00mmol) in MeOH (60mL) was added 1M K2C03 (20mL). After stirring for 12h, the reaction mixture was evaporated in vacuo, the residue was dissolved in H20 the pH was adjusted to ca. 6 with IM HC1, and the mixture was extracted with CHC13 (2X0.1L). The combined extracts were washed with H20, brine, dried (Na2SO4), and evaporated in vacuo to afford the title compound (1.89g) as a white solid. MP: 95-96°C WO 99/67230 WO 9967230PCTIUS99/1 4233 -124- (lit. 98- 10 1 I H-NMR: (300MHz, CDC13): 8 7.30-7.3 7 (5 4.00 (1 3.68 (1 H), 3.45 (1 3.20 (1 2.59 (1 2.46 (1 1.76-2.21 (4 1.31 (9 El/MS, m/z (rel. intensity):218 (base), 91 IR (nujol): 1719, 1497, 1451, 1367, 1296, 1285, 1260, 1160, 1153, 1079, 965, and 757cm- 1 Anal: Calcd. for C18H25N10 4 67.69; H, 7.89; N, 4.39. Found: C, 67.55; H, 7.97; N, 4.15; [xID 2 5 ±270 (c 0.795, CH2C12).
Preparation 27.
(Scheme E, E-6 where R 5 is 2 6 -dichlorophenyl)methoxyjphenyl, and stereochemnistry of the pyrrolidine ring is (2R,5S) and the amino acid is (1 S)-l -Methoxycarbonyl-2- [4-1 (2,6dichlo rophenyI)methoxyl phenyll ethyl] amino] 2oxoethyl 1 (phenylmethyl)-Lproline 1,1-dimethylethyl ester (IS, 5R, L)) c1 0 N ci 0 To a stirring solution of E-4 (Scheme E, where stereochemnistry is (2R,5S)) (0.48g, I .50mm~l) in CH2C12 (lOmL) was added 1- (3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (O.29g, 1 .50mmol), 1 -hydroxybenzotriazole hydrate (0.20g, 1 .50mmol), 4 -dimethylaminopyridine (0.05g, 0.45mmol), and 2,6dichlorobenzyl-L-tyrosine methyl ester hydrochloride (Scheme E, E-5: where R, is 4-[ (2,6-dichlorophenyl)methoxylphenyl and stereochemnistry is (0.59g, 1 .50mmol) to give a heterogeneous mixture. Upon addition of triethylamnine (0.3mL) the reaction WO 99/67230 PCT/US99/14233 -125mixture became homogeneous and stirred for 12h. The reaction mixture was partitioned between CH2C12 (50mL) and IN HC1 (50mL). The organic phase was washed with sat'd aq. NaHCO3, H20, brine, dried (Na2SO4), filtered and evaporated in vacuo. The resulting yellow oil was chromatographed on silica gel (150g, 230-400 mesh, 70mm OD column, packed and eluted acetone/CH2C12 5:95, 40mL fractions). Fractions 36-48 furnished the title compound (0.90g) as a glass. 1 H-NMR: (300MHz, CDC13): 8 9.09 (1 7.38 (2 7.10-7.30 (8 6.97 (2 5.22 (2 4.75 (1 3.81 (1 3.68 (3 H), 3.60 (1 3.15-3.35 (3 3.04 (1 2.42 (1 2.21 (1 1.86-2.07 (4 1.37 (9 H); FAB/MS m/z (rel. intensity): 655 46.4), 599 553 260 204 91 (base); IR (nujol):3262, 3001, 1733, 1665, 1612, 1585, 1565, 1512, 1439, 1392, 1240, 1226, 1197, 1177, 1153, 1018, and 768 cm- 1 Anal: Calcd. for C35H40N206C12: C, 64.12; H, 6.15; N, 4.27; Cl, 10.82. Found: C, 63.75 H, 6.29; N, 4.11; Cl, 10.88; [Ia]D 2 5 (c 0.863, CHC13).
Preparation 28.
(Scheme E, E-7 where R 5 is 2 6 -dichlorophenyl)methoxy]phenyl, and stereochemistry of the pyrrolidine ring is (2R,5S) and the amino acid is (1S)-l-Carboxy-2-14-[ (2,6dichlorophenyl)methoxy]phenyllethyl]amino]-2-oxoethyll-- (phenylmethyl)-Lproline 1,1-dimethylethyl ester 5R, L)) is 0
OH
o o o 0 N 0 To a stirring solution of E-6 (Scheme E, where R, is (2,6-dichlorophenyl)methoxy]phenyl, and stereochemistry of the pyrrolidine ring is (2R,5S) and the amino acid is WO 99/67230 PCT/US99/14233 -126- (1.00g, 1.53mmol) in MeOH (50mL) was added 1M K2CO3 (10mL). After stirring for 12h, the reaction mixture was evaporated in vacuo, the residue was dissolved in the pH was adjusted to ca. 6 with 1M HC1, and the mixture was extracted with CHC13 (2X0.1L). The combined extracts were washed with H20, brine, dried (Na2SO4), and evaporated in vacuo to afford the title compound (0.85g) as a white solid.
MP: 80-83°C; 1 H-NMR: (300MHz, CDC13): 8 9.87 (1 7.35 (2 7.21-7.26 (7 H), 7.13 (2 6.94 (2 5.22 (2 4.64 (1 3.80 (1 3.55 (1 3.29-3.37 (3 3.04 (1 2.48 (1 2.26 (1 1.94 (1 1.76 (1 1.59 (1 1.41 (1 1.34 (9 H); FAB/MS m/z (rel. intensity): 641 45.8), 585 260 204 (base), 91 IR (nujol): 1732, 1642, 1612, 1585, 1565, 1534, 1511, 1439, 1240, 1230, 1196, 1178, 1153, 1018, 779, and 767cm- 1 HRMS: Calcd. for C 34
H
3 ,Cl 2 N0 6 641.2185.
Found. 641.2164; [a]D 2 5 (c 0.795, CHC13).
Preparation 29 and Example 94.
(Scheme E, E-8 where R, is (2,6-dichlorophenyl)methoxy]phenyl, and stereochemistry of the pyrrolidine ring is (2R,5S) and the amino acid is (1S)-l-Carboxy-2-[4-[ (2,6dichlorophenyl)methoxy]phenyllethyl]amino]-2-oxoethyl]-1- (phenylmethyl)-Lproline 5R, L)) oci Cl o HO O N OH 1- WO 99/67230 PCT/US99/14233 -127- To a solution of E-7 (Scheme E, where R, is 2 6 -dichlorophenyl)methoxy]phenyl, and stereochemistry of the pyrrolidine ring is (2R,5S) and the amino acid is (0.85g, 1.32mmol) in H20/n-PrOH 0.1L) was added HOAc (6mL), and the solution refluxed for 5h, then stirred at RT for 12h. Evaporation in vacuo afforded the title compound (0.78g) as a white solid. MP: 198-202 0 C; IH-NMR: (300MHz, DMSO): 6 =8.56 (1 7.56-7.59 (2 7.45-7.50 (1 7.25-7.29 (5 7.21 (2 6.96 (2 5.18 (2 4.41 (1 3.89 (1 3.74 (1 3.31 (1 3.03-3.09 (2 2.85 (1 2.18-2.34 (2 1.88-2.00 (1 1.63-1.84 (2 1.44-1.58 (1 FAB/MS m/z (rel. intensity): 585 21.6), 539 246 204 159 91 (base); IR (nujol): 3309, 3083, 3037, 3014, 1662, 1644, 1562, 1514, 1440, 1377, 1348, 1241, 1197, 1178, 1018, 998, 815, and 771cml; Anal: Calcd. for C 3 0
H
30
N
2 0,C1 2 0.38H 2 0: C, 60.83; H, 5.24; N, 4.73. Found: C, 60.83; H, 5.33; N, 4.69; Karl Fischer water: 0.42%.
Example (Scheme E, E-8 where R, is (2,6-dichlorophenyl)methoxy]phenyl, and stereochemistry of the pyrrolidine ring is (2S,5R) and the amino acid is (1S)-l-Carboxy-2-[4-I 2 6 -dichlorophenyl)methoxy]phenyl]ethyl]amino]- 2-oxoethyl]-1- (phenylmethyl)-D-proline 5S, D)) Cl o o Example 95 was prepared as described in Scheme E from E-1 (Scheme E where stereochemistry is prepared by the method ofRapoport Am. Chem. Soc. 1984, 106, 4539). Physical data as follows: 198-204 0 C; 1 H-NMR: (300MHz, DMSO): 8 8.60 (1 7.54-7.68 (3 7.26-7.41 (8 7.05 (2 5.27 (2 4.57 (1 3.96 (1 3.82 (1 WO 99/67230 PCTIUS99/1 4233 -128- 3.40 (1 3.14 (2 2.86-2.94 (1 2.27-2.47 (2 1.96-2. 10 (1 1.72-1.88 (1 1.55-1.72 (1 1.30-1.42 (1 FAD/MS m/z (rel. intensity): 585 50.5), 539 332 331 246 244 204 91 (base); IR (nujol): 3211, 3033, 3006, 1724, 1647, 1610, 1565, 1512, 1438,1354,1301,1273,1240,1196, 1018, 871, and 767cm'; Anal: Calcd. for C30H30N206C12-0.43H 2 0:C, 60.77; H, 5.24; N, 4.72.
Found: C, 60.76; H, 5.37; N, 4.59; Karl Fischer water analysis: 1.71%.
Scheme F S RF.1 RN OH2 F-i
SH
2 N RF-3 F-2 0 RF.1F- RF-2ls F 0F-72 SRF-
RF-
J RFRF1 tI(NHCKRF-3 -2 0F N NH~ HF5-4 Hj OHK 0 F H 0 R 5
F-
c: RF-I CSRF-l H ,F-3 N o H O RIY0 R 5 F5R Y0IRS F-6 Where RF- 3 is defined as proton or C 1 ,alkyl.
WO 99/67230 PCT/US99/14233 -129- Scheme F describes a general method for the preparation of examples of the formula F-4, F-5, F-6, F-7, and F-8. A commercially available or readily prepared sulfur containing amino acid of structure F-1 is condensed with amino acid derivative F-2 under standard peptide synthesis conditions as described in Scheme A. Deprotection of the carbamate from F-3 provides the useful intermediate F-4. The amine group may be reacted with a variety of electrophilic reagents as described in Scheme A to provide esters of general structure F-5. Mild base hydrolysis provides acids of structure F-6. Mild hydrolysis of esters of general structure F-3 provides acid of formula F-7. In those cases in which R.3 is equal to t-butyl, mild acidolysis of compounds of general structure F-3 afford the amino acid of general structure F-8.
Preparation 30 and Example 96.
(Scheme F, F-3: where RF., and RF- 2 are the same and equal to proton, RF.3 is CH 3 R, is 4- 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemistry is S)) ^0-S o c O"0o 0 CI c To a cooled (0 solution of Boc-L-thiomorpholine-3-carboxylic acid Van Der Auwera, Anteunis, M.J.O. Int. J. Peptide Protein Res. 1987, 29, 574: Kogami, Y.; Okawa, K. Bull. Chem. Soc. Jpn. 1987, 60, 2963: Larsson Carlson R. ACTA Chemica. Scand 1994, 48, 517: Carson Wong F.F. J. Org. Chem. 1964, 29, 2203.) (Scheme F, F-l: where RF, and RF.2 are the same and equal to proton and stereochemistry is (6.7 g, 27 mmol) in CH 2
C
2 (100 mL) was added HOBt (4.0 g, 29.7 mmol), DMAP (700 mg), EDC (5.7 g, 29.7 mmol) and triethylamine (13.5 mL, 97 mmol).
The reaction mixture was stirred for 10 min, then the amino acid derivative F-2 (Scheme F, where R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl,
RF
3 is CH 3 and stereochemistry is (10.0 g, 24.7 mmol) was added. After 20 h, volatiles were removed in vacuo and the residue partitioned between 2.5% aqueous HC1 (100 mL) and H 2 0 (100 mL). The organic WO 99/67230 PCT/US99/14233 -130layer was separated and washed saturated aqueous NaHCO 3 (100 mL), dried and concentrated in vacuo. Purification of the residue by chromatography on SiO 2 (500 g) using CH 2
CI
2 /ethyl acetate as eluent afforded the title compound (12.31 g) as a solid: 'H NMR (CDC1 3 8 1.44 (9 2.35 (1 2.70 (3 3.13 (2 3.33 (1 3.77 (3 4.22 (1 5.00 (1 6.48 1 7.18 (2 7.31 (3 7.44 (1 7.56 (2 H); 3 C NMR (CDC1 3 171.6, 168.9, 162.5, 136.5, 135.9, 132.4, 131.0, 130.2, 128.2, 120.5, 81.7, 77.3, 53.3, 52.6, 37.0, 28.2, 26.5; IR (mull) 3296, 2924, 1744, 1685, 1668, 1605, 1536, 1515, 1432, 1412, 1321, 1294, 1260, 1244, 1213, 1195, 1161, 798 cm'; MS (FAB) m/z (rel. intensity) 598 596 Anal. Calcd for C 2 7
H
31 Cl 2
N
3 0S C, 54.36; H, 5.24; N, 7.04. Found: C, 54.23; H, 5.24; N, 6.86. Corrected for 0.60% found by Karl Fischer analysis.
Preparation 31 and Example 97.
(Scheme F, F-4: where and RF.
2 are the same and equal to proton, RF.
3 is CH 3 Rs is 4- [(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is S)) o H 0
N
Acetyl chloride (1.75 mL, 24 mmol) was slowly added to MeOH (26 mL) at 0-5 OC. After min, a solution of the carbamate F-3 (Scheme F, where RF., and RF, are the same and equal to proton, RF.3 is CH 3 R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (2.4 g, 4.0 mmol) in methanol (8 mL) was added. After 50.5 h at 0 the solvent was removed in vacuo to yield the title compound (2.11 'H NMR (300 MHz, DMSO-d 6 8 2.68 (1 3.00 (6 3.47 (1 3.64 (3 4.02 (1 4,52 (1 7.28 (2 7.54 (5 9.15 (1 9.3 (1 9.70 (1 10.7 (1 IR (mull) 3191, 3031, 1742, 1664, 1604, 1577, 1561, 1540,1516, 1432, 1414, 1326, 1271, 1210, 799 cm MS (EI) m/z (rel. intensity) 495 1).
WO 99/67230 PCT/US99/14233 -131- Preparation 32 and Example 98.
(Scheme F, F-5: where RF, and RF, are the same and equal to proton, R,.
3 is CH,, R 3 is
CH
2
CH,CO
2
CH
3
R
5 is 4 2 ,6-dichlorobenzoyl)amino]phenyl, Y is CO-, and stereochemistry is s O 0o 0 V0 0 o ci To a solution of amine F-4 (Scheme F, where RF,- and RF.2 are the same and equal to proton, RF.3 is CH 3 R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (R, (650 mg, 1.2 mmol) was added mono-methyl succinate (320 mg, 2.4 mmol), EDC (460 mg, 2.4 mmol), pyridine (10 mL) and DMAP at ambient temperature. After 27 h, the mixture was diluted with 25 mL of saturated NaHCO 3 was extracted with methylene chloride. The combined organic extracts were dried and concentrated in vacuo.
Purification of the residue by flash chromatography using methylene chloride/ethyl acetate as eluant followed by lyophilization afforded the title compound (600 mg) as an amorphous solid: IR(mull) 1742, 1659, 1657, 1608, 1537, 1516, 1432, 1414, 1324, 1269, 1259, 1228, 1214, 1195, 1176 'H NMR (300 MHz, CDCl 3 8 2.63 (7 3.21 (3 H), 3.68 (3 3.78 (3 3.90 (1 4.80 (2 5.50 (1 6.56 (1 7.29 (5 7.57 (2 'C NMR (75 MHz, CDCl 3 6 173.9, 172.0, 171.7, 168.5, 162.7, 136.1, 135.9, 133.3, 132.3, 131.0, 130.1, 128.2, 121.0, 120.7, 53.1, 52.5, 52.3, 52.0, 44.4, 36.7, 29.4, 27.7, 26.9, 26.4; MS (EI) m/z (rel. intensity) 609 Anal. Calcd for C 27
H
29 C1 2
N
3 0 7
S
C, 53.12; H, 4.79; N, 6.88. Found: C, 53.04; H, 4.81; N, 6.83. Corrected for 0.74%
H
2 0 found by Karl Fischer analysis.
Preparation 33 and Example 99.
3-[[[1-Carboxy-2-[4-[(2,6dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-y-oxo-4-thiomorpholinebutanoic acid WO 99/67230 PCT/US99/14233 -132- (Scheme F, F-6: where RF, and R, 2 are the same and equal to proton, RF.
3 is proton, R 3 is
CH
2
CH
2 COH, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl, Y is CO-, and stereochemistry is 0 N yNH AOH HO
CI
ooc 0 IN H To a solution of the diester F-5 (Scheme F, where and RF.
2 are the same and equal to proton, RF.
3 is CH 3
R
3 is CH 2
CH
2
CO
2
CH
3 R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl,
Y
is CO-, and stereochemistry is (490 mg, 0.80 mmol) in THF (20 mL) and MeOH (6 mL) was added a solution of LiOH*H 2 0 (178 mg, 4.25 mmol) in H 2 0,O (6 mL). After 22 h, the mixture was concentrated in vacuo. The residue was partially dissolved in 10% HCI mL) and the resulting solid collected by filtration. The solid was washed with water and lyophilized from aqueous acetonitrile to afford the title compound (400 mg) as an amorphous solid: IR (mull) 3267, 3193, 3058, 3034, 2924, 1725, 1658, 1607, 1562, 1537, 1516, 1432, 1414, 1326, 1195, 1178, 800 'H NMR (300 MHz, DMSO-d 6 8 2.62 (7 3.17 (3 3.95 (1 4.57 (2 5.26 (1 7.18 (2 7.55 (5 8.02 (1 H), 10.64 (1 12.34 (1 MS (FAB) m/z (rel. intensity) 582 18).
Preparation 34 and Example 100.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-4- thiomorpholinecarboxylic acid 4-(1,1 -dimethylethyl) ester (Scheme F, F-7: where and RF- 2 are the same and equal to proton, R, 3 is proton, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -133- S N O CN' NH C I To a solution of ester F-3 (Scheme F, where and RF-2 are the same and equal to proton,
R,.
3 is CH 3 Rs is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is S)) (656 mg, 1.02 mmol) in MeOH (25 mL) was added K2CO 3 (550 mg, 4 mmol) and H 2 0 (13 mL). After 3 h, volatiles were partially removed and the solution diluted with 10% HCI mL) causing precipitation of a solid. The product was collected by filtration, washed with H 2 0 and dried in a vacuum oven to afford the product (610 mg): IR (mull) 1736, 1665, 1606, 1562, 1537, 1516, 1432, 1413, 1323, 1294, 1260, 1244, 1211, 1195, 1160 cm 'H NMR (300 MHz, DMSO-d 6 6 1.35 (9 2.45 (2 2.74 (5 4.00 (1 4.47 (1 4.70 (1 7.19 (2 7.52 (5 7.92 (1 10.60 (1 12.75 (1 MS (FAB) m/z (rel. intensity) 582 12). Anal. Calcd for C 2 6
H
2 9 C1 2
N
3 0S C, 53.61; H, 5.02; N, 7.21;. Found: C, 53.20; H, 5.12; N, 7.10. Corrected for 2.30% H 2 0 found by Karl Fischer analysis.
Preparation (Scheme F, F-3: where RF. and RF.
2 are the same and equal to proton, R.
3 is t-butyl, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is o "Y o c To a cooled (0-5 solution of Boc-L-thiomorpholine-3-carboxylic acid (Scheme F, F- 1: where RF., and RF.
2 are the same and equal to proton and stereochemistry is (1.34 g, 5.4 mmol) in methylene chloride (20 mL) was added HOBt (800 mg, 5.94 mmol), DMAP WO 99/67230 PCT/US99/14233 -134- (140 mg, EDC (1.14 g, 5.94 mmol) and triethylamine (2.7 mL, 19.4 mmol). After min, F-2 (Scheme F where and RF.
2 are the same and equal to proton, R_ 3 is t-butyl, R is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (2.02 g, 4.94 mmol) was added, the reaction allowed to warm to ambient temperature and stirred for 24 h.
Volatiles were removed in vacuo and the residue partitioned between methylene chloride and 2.5% aqueous HC1. The organic layer was separated and washed with sat. aqueous NaHCO 3 dried and concentrated in vacuo. Purification of the residue by flash chromatography using methylene chloride/ethyl acetate as eluant afforded the title compound (1.64 IR (mull) 1730, 1687, 1667, 1606, 1538, 1515, 1431, 1412, 1395, 1320, 1294, 1258, 1250, 1194, 1158 cm'; 'H NMR (300 MHz, CDCI 3 8 1.44 (9 1.46 (9 2.35 (1 2.67 (3 3.22 (3 4.25 (1 4.73 (1 4.97 (1 6.52 (1 H), 7.29 (5 7.53 (3 MS (FAB) m/z (rel. intensity) 638 Anal. Calcd for
C
3 0
H
3 7 Cl 2 N30 6 S: C, 56.42; H, 5.84; N, 6.58. Found: C, 56.13; H, 5.98; N, 6.58.
Preparation 36 and Example 101.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(3R)-3-thiomorpholinyl]carbonyl]-Lphenylalanine monohydrochloride (Scheme F, F-8: where RF., and RF- 2 are the same and equal to proton, R.
3 is proton, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is S O N O NH H o 0 Cl To a solution of HCI saturated in ethyl ether (5 mL) at ambient temperature was added carbamate F-3 (Scheme F, where RF,_ and RF-2 are the same and equal to proton, RF.
3 is tbutyl, Rs is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (100 mg, 0.15 mmol) with vigorous stirring. After 43.5 h, the precipitate was collected by filtration and washed with ethyl ether to afford the title compound (90 mg): IR (mull) 3241, 3189, 3033, 2731, 1725, 1661, 1605, 1578, 1562, 1542, 1515, 1432, 1414, 1328, 1195 cnm'; 'H WO 99/67230 PCT/US99/14233 -135- NMR (300 MHz, DMSO-d 6 8 2.66 (1 2.99 (6 3.50 (1 3.98 (1 4.45 (1 H), 7.26 (1 7.52 (5 9.00 (2 9.30 (1 10.69 (1 MS (FAB) m/z (rel. intensity) 482 83), 540 539 538 486 485 484 483 482 173 102 Anal. Calcd for C 2
,H
2 ,C1,N 3 0 4 S HCl: C, 48.61; H, 4.27; N, 8.10; Cl, 20.50; S, 6.18. Found: C, 48.92; H, 4.27; N, 7.79; Cl, 19.68. Corrected for 6.53% H,O found by Karl Fischer analysis.
Example 102.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(3R)-4-[ 1 -oxo-3-(l H-tetrazol-5-yl)propyl]-3thiomorpholinyl]carbonyl]-L-phenylalanine methyl ester (Scheme F, F-5: where RF,, and RF., are the same and equal to proton, RF.
3 is CH 3
R
3 is 2- (5-1 H-tetrazolyl)ethyl, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl, Y is CO-, and stereochemistry is Hs H 0 N 0 0 CI$.
1 0C
N-N
Example 102 was prepared as described in Scheme F using 1H-tetrazole-5-propanoic acid (Hutchinson, Naylor, M. Nucleic Acids Res. 1985, 13, 8519) to form the requisite amide. Physical data as follows: IR (mull) 3264, 3047, 1742, 1659, 1607, 1561, 1537, 1516, 1432, 1415, 1324, 1268, 1219, 1195, 799 cm'; 'HNMR(300 MHz, DMSO-d6) 6 2.26 (1 2.66 (2 3.02 (8 3.61 (3 3.87 (1 4.56 (1 5.25 (1 7.21 (2 7.50 (5 8.18 (1 8.45 (1 8.64 (1 MS (FAB) m/z (rel. intensity) 620 61).
Example 103.
4 -[(2,6-Dichlorobenzoyl)amino]-N-[[(3R)-4-[1 -oxo-3-( I H-tetrazol-5-yl)propyl]-3thiomorpholinyl]-carbonyl]-L-phenylalanine WO 99/67230 WO 9967230PCTJUS99/I 4233 -136- (Scheme F, F-6: where RF- 1 and R- 2 are the same and equal to proton, RF.
3 is proton, R 3 is 2-(5-1H-tetrazolyl)ethyl, R, is 4-Ij(2,6-dichlorobenzoyl)amino]phenyl, Y is CO-, and stereochemnistry is 0
HH
N 0 1
I
NN N
H
Cl Example 103 was prepared from example 102 by the procedure described in preparation 34. Physical data as follows: IR (mull) 3376, 3296, 3267, 3127, 1746, 1683, 1669, 1641, 1623, 1610, 1542, 1522, 1444, 143 6, 1411 cm' MS (ESI+) for C 25
H
25 C1 2
N
7 0 5 S m/z 605.8 MS (FAB) m/z (rel. intensity) 606 (MW, 51), 682 608 607 606 605 254 226 175 137 102 HRMS (FAB) calcd for
C,
5
H
2 5 C1 2
N
7 0 5 5 +Hl 606.1093, found 606.1105.
Example 104.
R)-4-(3-Cyano-lI-oxopropyl)-3-thiomorpholinyl]carbonyl]-4-[[2,6dichlorobenzoyl)aminol-L-phenylalanine (Scheme F, F-6: where RF-I and RF.2 are the same and equal to proton, RF-A is proton, R 3 is
CH
2
CH
2 CN, R 5 is 4-[(2,6-dichlorobenzoyl)aminolphenyl, Y is CO-, and stereochemnistry is WO 99/67230 WO 9967230PCTIUS99/1 4233 -137-
H
(N
N
N OH NC0 0 C I
N
H
rCl1 Example 104 was prepared as described in Scheme F using 3-cyanopropanoic acid (readily prepared from commercially available 3 -cyanopropanoic acid) to form the requisite amide.
Physical data as follows: IR (mull) 2251, 1735, 1655, 1612, 1585, 1565, 1512, 1439, 1298, 1240, 1196, 1179, 1016, 1000, 779, 768 cm- 1 'H NMR (300 MHz, CDCI 3 )8~2.40 (12 4.88 (1 5.22 (2 6.72 (1 6.96 (2 7.23 (5 H).
Example 105.
1 -Carboxy-2-[4-[(2,6-dichlorophenyl)methoxy].
phenyl]ethyl]amino]carbonyl]py.ox..{..-thiomorpholinebutanoic acid (Scheme F, F-6: where RF-, and RF.
2 are the same and equal to proton, RF.
3 is Proton., R 3 is
CH
2
CH
2
CO
2 H, R, is 4 2 6 -dichlorophenyl)methoxylphenyl Y is CO-, and stereochemnistry is
S
0 C HN OH HO0 0 -Cl Example 105 was prepared as described in Scheme F using mono-methyl succinate to form the requisite amide. Physical data as follows: IR (mull) 3031, 1726, 1646, 1612, 1585, 1565, 1511, 1439, 1421, 1297, 1240, 1196, 1179, 1016, 768 cm-1; 'H NMR (300 MHz, WO 99/67230 PCT/US99/14233 -138- DMSO-d 6 8 2.62 (8 3.64 (3 4.39 (2 5.20 (2 6.92 (2 7.15 (2 7.50 (3 7.98 (2 "C NMR (75 MHz, DMSO-d 6 6 174.5, 173.2, 171.9, 168.9, 136.5, 132.3, 132.0, 130.8, 130.7, 129.2, 114.8, 65.3, 54.3, 52.5, 36.0, 29.7, 28.0, 27.0; MS (FAB) m/z (rel. intensity) 569 24). Anal. Calcd for C 25
H
26
C
2
N
2 0 7 S: C, 52.73; H, 4.60; N, 4.92; Cl, 12.45. Found: C, 52.51; H, 4.60; N, 4.94; Cl, 12.78. Corrected for 3.37% H0O found by Karl Fischer analysis.
WO 99/67230 WO 9967230PCT/US99/1 4233 -139- Scheme G 0 0 NH g o RG-1 0 o N-RG.
3 RG-2- 9 0 o RG.1 G-1 G-2
H
2 N
O
0 G-3 G-4 G-6 Where and 2 are defined independently as H or CH 3
R-
3 is defined as H, C 1 -6alkyl or C 3 6 alkenyl; and g is defined as 0 or 2.
WO 99/67230 PCT/US99/14233 -140- Scheme G describes a general method to prepare lactam examples of general structures G-5 and G-6. Readily prepared lactams of general structure G-1 may be alkylated by the reaction of an appropriate alkylating in the presence of a suitable base as described in preparation 37 to provide intermediates of general structure G-2. Mild hydrolysis provides the monoacid of general structure G-3 which may be condensed with an amino acyl intermediate of structure G-4 as described in Scheme A. Full hydrolysis of the diester of general structure G-5 affords the diacid of structure G-6.
Preparation 37.
(Scheme G, G-2: where R o .G and R-2 are CH 3 R is CH 3 and g is equal to 2).
0
N-CH
3 0 0 To a cooled (0-5 solution of dimethyl ester G-1 (Scheme G, where RG, and R-2 are CH,, R,.3 is H and g is equal to 2) (Thomas, Rynbrandt, R.H; Zimmermann, D.C.; Bell, Muchmore, Yankee, E.W. J. Org. Chem. 1989, 54, 4535) (25.7 g, 0.1 mole) and iodomethane (30 mL, 0.48 mol) in THF (200 mL) was added NaH (4.8 g, 0.12 mmol, 60% in oil dispersion). After 22 h, the reaction was quenched by the addition of
H
2 0 (100 mL) and diluted with CH 2 Cl 2 (50 mL). The organic layer was separated, dried and dried in vacuo. The crude brown oil was triturated with hexanes (200 mL) and the residue concentrated in vacuo to afford the crude product (18.44 g) which was used without further purification: 'H NMR (300 MHz, CDCI 3 8 2.03 (12 2.64 (3 3.64 (6 H).
Preparation 38.
(Scheme G, G-3: whereR 2 is CH 3 RG. is CH 3 and g is equal to 2).
0
N-CH
3
O
0- -OH 0 0 WO 99/67230 PCT/US99/14233 -141- To a cooled (0-5 solution of diester G-2 (Scheme G, where RG., and R.
2 are CH 3
R-
is CH 3 and g is equal to 2) (10.0 g, 36.9 mmol) in aqueous methanol 75 mL) was added LiOH*H20 (1.55 g, 36.9 mmol). After 22 h, the mixture was partially concentrated in vacuo and diluted with water. The aqueous layer was washed with methylene chloride and acidified (pH ca. 2) with 10% HC1. The aqueous layer was extracted with methylene chloride and the combined organic extracts dried and concentrated in vacuo to afford the title compound (3.63 g) as a yellow solid: IR (mull) 1735, 1630, 1442, 1429, 1418, 1405, 1330, 1308, 1287, 1255, 1217, 1185, 1122, 1016, 642 cm'; 'H NMR (300 MHz, CDCl 3 1.90 (6 2.24 (4 2.43 (2 2.68 (3 3.67 (3 3 C NMR (75 MHz, CDCI 3 8 176.3, 175.5, 173.2), 64.6, 52.0, 33.1, 32.9, 29.9, 28.41, 28.38, 26.0, 24.9; MS (El) m/z (rel. intensity) 257 1).
Preparation 39 and Example 106.
2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]-3-oxopropyl]-1-methyl-5-oxo-2-pyrrolidinepropanoic acid methyl ester (Scheme G, G-5: where RG 2 and R.
3 are both equal to CH 3
R
5 is dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemistry is
N-CH
3 i0 0 0 ci To a solution of acid G-3 (Scheme G, whereRG 2 and RG 3 are equal to CH 3 and g is equal to 2) (1.0 g, 3.9 mmol), amine G-4 (Scheme G, where R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is (1.82 g, 4.67 mmol), and DMAP (100 mg, 0.8 mmol) in pyridine (15 mL) was added EDC (895 mg, 4.67 mmol). After 21 h, the reaction was diluted with saturated aqueous NaHCO 3 and methylene chloride. The organic WO 99/67230 PCT/US99/14233 -142layer was separated and washed with 10% aqueous HC1, dried and concentrated in vacuo.
Purification of the residue by flash chromatography using methylene chloride/ethyl acetate as eluant followed by lyophilization from aqueous acetonitrile afforded the title compound (1.71 g) as an amorphous solid: IR (mull) 3284, 2924, 1738, 1666, 1665, 1564, 1539, 1511, 1439, 1398, 1299, 1240, 1198, 1178, 1117, 1016, 768 'H NMR (300 MHz, CDC13) 8 2.10 (12 2.66 (3 3.07 (2 3.66 (3 3.73 (3 4.83 (1 H), 5.24 (2 5.98 (1 6.95 (2 7.02 2 7.25 (1 7.36 (2 3 C NMR (75 MHz,
CDCI
3 174.7, 174.66, 173.2, 172.0, 171.2), 158.1, 137.0), 132.1, 130.5, 130.3, 128.5, 128.3, 115.2, 65.2, 64.3, 53.3, 52.4, 51.9, 37.0, 33.2, 30.2, 30.17, 28.4, 26.0, 24.7; MS (EI) m/z (rel. intensity) 592 Anal. Calcd for C2 9
H
3 4 Cl 2
N
2 C, 58.69; H, 5.77; N, 4.72; Cl, 11.95. Found: C, 58.33; H, 5.65; N, 4.76; Cl, 11.89.
Preparation 40 and Example 107.
S)-1 -Carboxy-2-[4-[(2,6-dichlorophenyl)methoxy]phenyl]ethyl]amino]-3oxo-propyl]-1 -methyl-5-oxo-2-pyrrolidinepropanoic acid (Scheme G, G-6: where R.
3 is equal to CH 3
R
5 is dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemistry is 0
N-CH
3
O
HO-\ N Q)OH o o -cl To a solution of diester G-5 (Scheme G, where RG 2 and R, 3 are both equal to CH 3
R
5 is 4- [(2,6-dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemistry is (1.0 g, 1.68 mmol), in THF (30 mL) and MeOH (9 mL) was added LiOH-H 2 0 (370 mg, 8.8 mmol). After 22.5 h, the reaction mixture was acidified with 10% aqueous HC1 (30 mL) causing precipitation of a solid. The solid was collected by filtration and lyophilized from aqueous acetonitrile to afford the title compound (0.91 g) as an amorphous solid: IR (mull) 3031, 2925, 1727, 1637, 1585, 1564, 1543, 1511, 1439, 1424, 1404, 1299, 1240, WO 99/67230 PCT/US99/14233 -143- 1196, 1179, 768 cm'; 'H NMR (300 MHz, DMSO-d 6 S 1.77 (12 2.76 (1 3.97 (1 4.36 (1 5.16 (2 6.94 (2 7.15 (2 7.49 (3 8.16 (1 'C NMR MHz, CDC1 3 8 174.6, 174.1, 173.6, 172.2, 157.6, 136.5, 132.2, 132.0, 130.7, 130.6, 129.2, 114.7, 65.3, 64.1, 54.0, 36.5, 33.7, 33.1, 30.1, 30.05, 29.8, 28.7, 25.7, 24.6; MS (FAB) m/z (rel. intensity) 565 99). Anal. Calcd for C 27
H
30 C1 2
N
2 C, 57.35; H, 5.35; N, 4.95; Cl, 12.54. Found: C, 56.93; H, 5.15; N, 5.02; Cl, 12.42. Corrected for 1.03% H 2 0 found by Karl Fischer analysis.
Example 108.
4 2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2oxoethyl]amino]-3-oxopropyl]--( 3 -methyl- 2 -butenyl)-5-oxo-2-pyrrolidinepropanoic acid methyl ester (Scheme G, G-5: where RG-2 is CH 3 R.3 is 1-(3-methyl-2-butenyl), R, is dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemistry is 0 0 ciD Example 108 was prepared as described in Scheme G using 1-bromo-3-methyl-2-butene to form the requisite N-alkyl lactam. Physical data as follows: IR (mull) 32.95, 29.52, 1740, 1678, 1662, 1564, 1538, 1512, 1439, 1414, 1300, 1279, 1240, 1199, 1178, 1017, 768 cm*'; 'H NMR (300 MHz, CDCI 3 8 1.64 (3 1.70 (3 2.06 (12 3.09 (2 3.66 (3 H), 3.74 (2 3.74 (3 4.85 (1 5.20 (1 5.24 (2 6.95 (2 7.02 (2 7.25 (1 7.36 (2 'C NMR (75 MHz, CDCl 3 8 174.7, 173.3, 172.1, 172.0, 171.3, 158.1, 137.0, 135.1, 132.1, 130.5, 130.3, 128.5, 128.3, 119.7, 115.2, 65.2, 65.1, 53.2, 52.4, 51.9, 37.5, 37.0, 34.4, 34.2, 30.6, 29.9, 28.6, 26.6, 25.6, 25.5, 17.9; MS (FAB) m/z (rel.
intensity) 647 24).
WO 99/67230 WO 9967230PCTIUS99/1 4233 -144- Example 109.
1S)-lI-Carboxy-2-[4-[(2,6-dichlorophenyl)methoxy]phenyl]ethyl]amino-3oxo-propyl]-1 -(3-methyl-2-butenyl)-5-oxo-2-pyrrolidinepropanoic acid (Scheme G, G-6: where RkJs 1 -methyl-2-butenyl), R, is 4+(2,6dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemnistry is 0 NL 0 HO O C4 Example 109 was prepared from example 108 by the procedure described in preparation 40. Physical data as follows: IR (mull) 3290, 2921, 1726, 1635, 1585, 1565, 1545, 1511, 1439, 1419, 1341, 1299, 1240, 1197, 1179, 780, 768 'H NMR (300 MHz, DMSOd6) 6 1.57(3 1.62 (3 1.72 (6 2.02 (6 2.77 (1 2.98 (1 3.55 (2 H), 5.05 (1 5.16 (2 6.94 (2 7.15 (2 7.49 (2 7.54 (2 8.15 (1 1 3
C
NMR (75 MHz, DMSO-d 6 5 174.6, 174.3, 173.6, 173.56, 172.2, 157.6, 136.5, 134.0, 132.2, 132.0, 130.6, 129.2, 121.1, 121.0, 114.7, 65.2, 65.0, 54.1, 54.0, 37.1, 36.5, 34.5, 34.1, 30.0, 29.9, 28.8, 26.3, 25.84), 25.81, 18.1; MS (FAB) m/z (rel. intensity) 619 (M+H, 99).
Example 1 1S)-I 4 6 -Dichlorobenzoyl)amino]phenyllmethyl]-2-methoxy-2oxehlaio--xpoy]lmthl5oo2proiierpni acid methyl ester (Scheme G, G-5: where k- 2 and k.3 are equal to CH 3 R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, g is equal to 2 and stereochemnistry is WO 99/67230 PCT/US99/14233 -145- 0
N-CH
3 0 0 0 0 c Example 110 was prepared as described in Scheme G using iodomethane to form the requisite N-alkyl lactam. Physical data as follows: IR (mull) 3258, 2922, 1738, 1662, 1606, 1561, 1539, 1515, 1432, 1414, 1401, 1323, 1268, 1196, 1177, 799 cm-; 'H NMR (300 MHz, CDC13) 5 2.01 (12 2.61 (3 3.09 (2 3.64 (3 3.75 (3 4.84 (1 6.15 (1 7.09 (2 2.31 (3 7.58 (2 7.99 (1 H);MS (EI) m/z (rel. intensity) 607 605 Anal. Calcd for C2 9
H
33 C1 2
N
3 0 7 C, 57.43; H, 5.48; N, 6.93; Cl, 11.69. Found: C, 57.18; H, 5.56; N, 6.85; Cl, 11.68. Corrected for 0.93% H 2 0, found by Karl Fischer analysis.
Example 111.
2-[3-[[(1S)-l-Carboxy- 2 4 -[(2,6-dichlorobenzoyl)amino]phenyl]ethyl]amino]-3-oxopropyl]-l-methyl-5-oxo-2-pyrrolidinepropanoic acid (Scheme G, G-6: where RG 3 is CH 3 R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, g is equal to 2 and stereochemistry is N-cH 3 0 HCO O Example 111 was prepared from example 110 by the procedure described in preparation 40. Physical data as follows: IR (mull) 3265, 3056, 2925, 1724, 1658, 1609, 1579, 1561, 1542, 1516, 1432, 1414, 1327, 1271, 1217, 1195, 800 cm'; 'H NMR (300 MHz, DMSO- WO 99/67230 WO 9967230PCTfUS99/1 4233 -146d6) 8 1.89 (12 2.49 (3 2.78 (1 2.99 (1 4.38 (1 7.18 (2 7.51 (5 H), 8.17 (1 10.64 (1 MIS (FAB) m/z (rel. intensity) 580.5 68).
Example 112.
1S)-i 4 2 6 -Dichlorophenyl)methoxylphenyllmethyl]-2-methoxy-2-.
oxoethyl]amino]-3-oxopropyll--5-oxo-2-pyrrolidinepropanoic acid methyl ester (Scheme G, G-5: where RG- 2 is CH 3
RG-
3 is proton, R5 is 4-[2,6-dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemistry is 0 NH 0 IppH 0 00 Example 112 was prepared as described in Scheme G. Physical data as follows: IR (mull) 3276, 3029, 1738, 1686, 1564, 1538, 1511, 1439, 1299, 1279, 1239, 1197, 1178, 1016, 767, 'H NMR (300 MHz, CDCl 3 6 1.85 (6 2.24 (2 2.35 (4 3.03 (2 H), 3.66 (3 3.74 (3 4.82 (1 5.24 (2 6.50 (2 6.95 (2 7.05 (2 7.27 (1 7.37 (2 MIS (El) m/z (rel. intensity) 578 Anal. Calcd for
C
2
,H
32 C1 2
N
2 0 7 C, 58.04; H, 5.57; N, 4.83. Found: C, 57.93; H, 5.43; N, 4.97. Corrected for 1. 14 H 2 0 found by Karl Fischer analysis.
Example 113.
lS)-l -Carboxy- 2 4 -[(2,6-dichlorophenyl)methoxy]phenyl]ethyllamino]-3 oxo-propyl]-S-oxo-2-pyrrolidinepropanoic acid (Scheme G, G-6: where is proton, R5 is 4-[(2,6-dichlorophenyl)methoxy]phenyl, g is equal to 2 and stereochemistry is WO 99/67230 PCT/US99/14233 -147- 0 O H O 0 0 Example 113 was prepared from example 112 by the procedure described in preparation Physical data as follows: IR (mull) 3294, 3033, 1716, 1647, 1585, 1565, 1544, 1511, 1439, 1420, 1299, 1240, 1197, 1179, 768 cm'; 'H NMR (300 MHz, DMSO-d 6 3 1.60 (6 2.12 (6 2.78 (1 2.99 (1 4.36 (1 5.19 (2 6.97 (2 7.17 (2 H), 7.45 (1 7.58 (2 7.70 (1 8.22 (1 MS (FAB) m/z (rel. intensity) 551 (M+H, 99); Anal. Calcd for C 26
H
2 8 C1 2
N
2 C, 56.63; H, 5.12; Cl, 12.86; N, 5.08. Found: C, 56.28; H, 5.01; Cl, 13.08; N, 5.24. Corrected for 1.47% H 2 0 found by Karl Fischer analysis.
Example 114.
4 -[(2,6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]-3-oxopropyl]-5-oxo-2-pyrrolidinepropanoic acid methyl ester (Scheme G, G-5: where RG 2 is CH,, RG.
3 is proton, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, g is equal to 2 and stereochemistry is
NH
/H
0 0 0 0 CI Example 114 was prepared as described in Scheme G. Physical data as follows: 'H NMR (300 MHz, CDCl1) 8 1.81 (6 2.27 (6 3.10 (2 3.63 (3 3.75 (3 4.89 (1 H), 6.46 (1 6.58 (1 7.10 (2 7.26 (3 7.58 (2 8.14 (1 MS (FAB) m/z 592 (M+H) 568, 367, 349, 307, 278, 226, 194, 173.
WO 99/67230 PCT/US99/14233 -148- Example 115.
1 -Carboxy- 1 -[4-[(2,6-dichlorobenzolyl)amino]phenyl]ethyl]amino]-3oxopropyl]-5-oxo-2-pyrrolidinepropanoic acid (Scheme G, G-6: where R-3 is proton, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, g is equal to 2 and stereochemistry is 0 NH 0
HH
HO N~).OH Example 115 was prepared from example 114 by the procedure described in preparation Physical data as follows: IR (mull) 3272, 3195, 3121, 3063, 2953, 2923, 2868, 2855, 1715, 1659, 1608, 1579, 1561, 1541, 1516, 1456, 1432, 1414, 1377, 1367, 1326, 1271, 1221, 1195, 800 cm- 1 'H NMR (300 MHz, DMSO-d 6 8 1.63 (6 2.13 (6 2.79 (1 3.00 (1 4.38 (1 7.19 (2 7.51 (5 7.64(1 8.16 (1 10.58 (1 H), 12.37 (2 MS (FAB) m/z 564 546, 519, 335, 280, 194, 173. Anal. Calc'd for
C
26
H
27 C1 2
N
3 C, 55.33; H, 4.82; Cl, 12.56; N, 7.44. Found: C, 55.10; H, 4.76; Cl, 12.56; N, 7.36. Corrected for 2.49% H 2 0, found by Karl Fischer analysis.
Example 116.
4 2 6 -Dichlorophenyl)methoxy]phenyl]methyl]-2-methoxy-2- (Scheme G, G-5: where RG.
2 and RG 3 are equal to proton, R5 is 4-[(2,6-dichlorophenyl)methoxy]phenyl, g is equal to 0 and stereochemistry is WO 99/67230 WO 9967230PCT/US9914233 -149- 0
NH
HO 0 0
CI
Example 115 was prepared as described in Scheme G from 2-carboxy-5-oxo-2pyrrolidinepropanoic acid (Majer, Kajtar, Tichy, Blaha, K. Coil. Czech. Chem.
Commun. 1982, 47, 950). Physical data as follows: IR (mull) 3302, 1736, 1671, 1612, 1585, 1564, 1535, 1511, 1439, 1298, 1240, 1197, 1179,1016, 768 1H NMR (300 MHz, CDCl 3 8 2.12 (8 3.02 (2 3.65 (3 4.81 (1 5.21 (2 6.94 (2 7.07 (2 7.22 (1 7.35 (2 7.86 (1 8.34 (1 MIS (FAB) m/z (rel. intensity) 537 99).
Example 117.
S)-i -Carboxy- 2 -[-[(2,6-dichlorophenyl)methoxy]phenylethyl]amino..3 (Scheme G. G-6: where R 0 3 is proton, R, is 4 -Ii( 2 6 -dichlorophenyl)methoxy]phenyl, g is equal to 0 and stereochemnistry is 0
NH
HO_ H 0 O-
O
Example 116 was prepared from example 115 by the procedure described in preparation Physical data as follows: IR (mull) 3292, 3059, 3029, 1718, 1650, 1612, 1585, 1565, 1537, 1511, 1439, 1240, 1196, 1179, 768 cm';'HNMR (300 MHzCDCl 3 82.11 (8 H), WO 99/67230 WO 9967230PCTIUS99/1 4233 -150- 2.83 (1 3.20 (1 4.74 (1 5.24 (2 6.95 (2 7.15 (2 7.25 (1 7.36 (2 MS (FAB) m/z (rel. intensity) 523 99); Anal. Calcd for C 24
H
24 C1 2 N20 7
C,
55.08; H, 4.62; N, 5.35, Cl, 13.55. Found: C, 54.68; H, 4.66; N, 5.13: Cl, 13.70.
Corrected for 1.59% H 2 0 found by Karl Fischer analysis.
Scheme H.
H-i N CI OHH-2 I N CI
I
KY
0 CI H-3 I N CI H0 PN- CI C BN ci H-4 .2 HCI WO 99/67230 PCT/US99/14233 -151- Scheme H teaches a general method for the preparation of 6-chloroazatyrosine examples of structures H-4 and H-5, by adaption of the methodology for the preparation of protected azatyrosine reagents as described by Kawata, Ashizawa, Hirama, M. J. Am. Chem.
Soc. 1997, 119, 12012-12013 and references cited therein. Thus regioselective iodination of 2-chloro-3-pyridinol gives the chloro-iodopyridinol H-2, that is O-alkylated as exemplified by the synthesis of H-3. Palladium-catalyzed reaction of H-3 with the organozinc derived from a suitably protected (3-iodoalanine, provides the protected chloroazatyrosine H-4. N-deprotection of H-4 gives the aminoester H-5, that is used (as exemplified by reagent A-4 of Scheme A, and by reagent B-5 of Scheme B) for the synthesis of Examples of this invention Preparation 41.
(Scheme H: H-2) 2-Chloro-6-iodo-3-pyridinol
(C
5
H
3 ClINO).
To a solution of 2-chloro-3-pyridinol H-1 (10.2 g, 78.7 mmol) and K 2
CO
3 (38.9 g, 0.274 mol) in H 2 0 (120 mL) is added 12 (24.3 g, 95.8 mmol), and the reaction mixture is stirred at rt for 4 h. The reaction mixture is quenched by the addition of aq satd Na 2
S
2 03-5H 2 0, and its pH is lowered to pH 2 with the addition of 12 M aq HC1. The mixture is extracted with EtOAc. The combined EtOAc extracts are dried, filtered and concnetrated to a yellow solid, that is crystallized from 120:25 heptane/EtOAc (145 mL) to give, as a yellow solid, 11.2 g of the title compound: 1 H NMR (CD 3
SOCD
3 300 MHz) 5 9.87 7.59 7.06 13 C NMR (CD 3
SOCD
3 75 MHz) 5 150.68, 138.07, 134.98, 127.02, 101.18.
Preparation 42.
(Scheme H: H-3) 2 -Chloro-3-[(2,6-dichlorophenyl)methoxy]-6iodopyridine (C1 2
H
7 Cl3INO).
To a solution of H-2 (5.11 g, 20.0 mmol), PPh 3 (5.30 g, 20.0 mmol), and 2,6dichlorobenzylalcohol (3.54 g, 20.0 mmol) in dry THF (100 mL) at 0 oC under Ar is added WO 99/67230 PCT/US99/14233 -152- DEAD (3.15 mL, 20.0 mmol). The reaction mixture is kept at 0 °C for 1.5 h and at rt for h. It is concentrated to a residue, that is purified by silica flash chromatography (17:3 hexanes/EtOAc) to give 7.61 g of the title compound: TLC (17:3 hexanes/EtOAc) Rf0.5 7 1H NMR (CD 3
SOCD
3 300 MHz) 5 7.85 7.62 7.58-7.45 5.34 (2H).
Preparation 43.
(Scheme H: H-4) (S)-2-Chloro-3-[(2,6-dichlorophenyl)methoxy]-aH-[[(1,1-dimethylethoxy)carbonyllamino]-6-pyridinepropanoic acid methyl ester (C 2 1
H
2 3 C1 3
N
2 0 5 To an amberized flask containing activated Zn dust (0.777 g, 11.89 mmol) under Ar is added sequentially N-[(1,l-dimethylethoxy)carbonyl]-3-iodo-L-alanine methyl ester [93267-04-0] (3.91 g, 1.89 mmol), THF (11.9 mL), and dimethylacetamide (11.9 mL). The reaction mixture was purged of 02 by the bubbling of Ar through the mixture for 5 min. It is stirred at 65±5 oC for 2 h, and is cooled to rt. To this mixture is added PdCl 2 (PPh 3 2 (0.412 followed immediately afterward by a degassed solution of H-3 (2.46 g, 5.94 mmol) in 1:1 THF/dimethylacetamide (11.8 mL). The reaction mixture is stirred at 65±5 °C for 5 h. It is cooled to 0 °C and quenched with sat'd aq NH 4 Cl (100 mL). The reaction mixture is extracted with EtOAc. The combined extracts are washed with brine, dried, filtered and concentrated to a green-yellow oil; that is purified by silica flash chromatography to give 1.90 g of the title compound: TLC (7:3 hexanes/EtOAc) Rf 0.32; 1 H NMR (CDC13, 300 MHz) 8 7.76 7.57 7.48 7.29 7.27 5.30 4.32 3.60 3.01 2.98 1.31 (9H).
Preparation 44.
(Scheme H: H-4) (S)-2-Chloro-3-[(2,6-dichlorophenyl)methoxyl--amino-6- pyridinepropanoicacid methyl ester dihydrochloride salt (C 16
H
15
CI
3
N
2 0 3 "2HCI).
A solution of H-4 (1.90 g, 3.88 mmol) in 4 M HCI in dioxane (35 mL) is stirred at rt under Ar for 20 h. The reaction mixture is concentrated in vacuo. The residue is taken up in H 2 0 (40 mL), and extracted with Et20. The aqueous solution is frozen and lyophilized WO 99/67230 WO 9967230PCTIUS99/1 4233 -153to give 1.39 g of the title compound: 'H NMR (CD 3
SOCD
3 300 MHz) 8 8.62 7.81 7.58 7.48 7.38 5.32 4.37 3.72 3.27 (2H).
Example 118.
(Scheme A, 2 -Chloro-3-[(2,6-dichlorophenyl)methoxy-6.
pyridyll methyll-2-methoxy-2-oxoethyll amino] carbonylj-3-thiazolidinecarboxylic acid (1 ,1-dimethylethyl) ester (C 30
H
3 7 C1 3
N
3 0 6
S).
Example 118 was prepared as described in Scheme A from D-cysteine using the product of preparation 44 as amino acid intermediate A-4. Physical properties as follows: TLC 1 hexanes/EtOAc) RjO0.
2 7; 1 H NMR (CDCl 3 300 MHz) 8 8.43 (1 7.74 (1 H), 8 7.48 (1IH), 7.29 (1 5.29 4.67 (1 4.5 3 (1IH), 4.44 (1 4.23 (1 H), 3.62 3.06 2.82 1.27 (9H).
Example 119.
(Scheme A, A-9) IS-(R *,R*)J-4-[11-Carboxy-2-t12-Chloro--3-[(2,6--dichlorophenyl)methoxy1-6pyridyll ethyl] amino] carbonylj-3-thiazolidinecarboxylic acid (1 ,1-dimethylethyl) ester (C 24
H
2 6 C1 3
N
3 0 6
S).
Example 119 was prepared from example 118 by the procedure described in preparation 12. Physical data as follows: mnp 93-95 TLC (600:400:2 EtOAc/hexanes/HCO 2 H) Rj 0.38; 'H NMR (CD 3
SOCD
3 300 MHz) 6 8.30 7.74 7.57 7.48 (111), 7.27 5.29 4.67-4.36 4.53 4.23 3.37-3.11 2.97 (1H), 1.27 (9H).
WO 99/67230 WO 9967230PCTIUS99/1 4233 -154- Scheme 1.
I
O
I N CI 0 o BoC.. 0 1-3
N
H 0 Boc.0 0 BN Nl 1-4
H
0
NOH
N
BN.. 0 0HC
CI
N
0
CN
N 0 1-7 0 CI -C WO 99/67230 PCT/US99/14233 -155- Scheme I teaches a general method for the preparation of azatyrosine reagents 1-3, 1-4, 1-6 and 1-7, by adaptation of the methodology for the preparation of protected azatyrosine reagents as described by Kawata, Ashizawa, Hirama, M. J. Am. Chem. Soc. 1997, 119, 12012-12013 and references cited therein. Thus O-protection of chloroiodopyridinol I-1 (identical to H-2) is followed by reaction with the organozinc, derived from a suitably protected P-iodoalanine, to provide the protected 6-chloroazatyrosine 1-3. Reductive dehalogenation of 1-3 gives 1-4, that is O-deprotected to give 1-5. Reagent 1-5 is Oalkylated, as exemplified by the preparation of 1-6. N-deprotection of 1-6 gives the aminoester 1-7, that is used (as exemplified by reagent A-4 of Scheme A, and by reagent B-5 of Scheme B) for the synthesis of Examples of this invention.
Preparation (Scheme I, 1-2) 2 -Chloro-3-[(2-tetrahydropyranyl)oxyl-6-iodopyridine
(C
10 H 11ClINO 2 To a solution of chloroiodopyridinol I-1 (same as H-2, the product of preparation 41) (1.00 g, 3.91 mmol) and dihydropyran (1.0 mL, 10.6 mmol) in CH 2 Cl 2 (10 mL) under Ar at rt is added pyridinium chloride (0.050 The reaction mixture is stirred for 72 h. It is diluted with CH 2 C12, and is washed with satd aq NaHCO 3 and brine. The CH 2 Cl 2 solution is dried, filtered and concentrated to an oil, that is purified by silica flash chromatography (19:1 hexanes/EtOAc) to give 1.06 g of the title product: TLC (19:1 hexanes/EtOAc) Rf0.24; 1H NMR (CDC13, 300 MHz) 5 7.55 7.17 5.50 (1H), 3.77 3.61 2.07-1.57 (6H).
Preparation 46.
(Scheme I, I-3) (S)-2-Chloro--a-[[(11,l-dimethylethoxy)carbonyl]amino]-3-[(2-tetrahydropyranyl)oxy]-6-pyridinepropanoic acid methyl ester (C 19H 2 7 C1N 2 0 6 To an amberized flask containing activated Zn dust (0.349 g, 5.51 mmol) under Ar is added THF (2 mL) and 1,2-dibromoethane (0.018 mL, 0.21 mmol). The suspension is WO 99/67230 PCT/US99/14233 -156brought to reflux for several minutes, cooled to approximately 30 OC, and TMSCI (0.17 mL ofa 1 M solution in THF) is added. The reaction mixture is stirred at 40±5 °C for min and then is cooled to below rt. A solution of N-[(1,1-dimethylethoxy)carbonyl]-3iodo-L-alanine methyl ester [93267-04-0] (1.81 g, 5.50 mmol) in 11:7 dimethylacetamide/THF (9.0 mL) is added, and the resulting reaction mixture is stirred at °C for 5 h. The reaction mixture is cooled to below rt, and solid PdCl 2 (PPh 3 2 (0.192 g) is added, followed immediately by addition of a degassed solution of the iodopyridine (0.936 g, 2.76 mmol) in 1:1 THF/dimethylacetamide (5.6 mL). This reaction mixture is stirred for 4 h at 45 It is cooled to 0 quenched with sat'd aq NH 4 CI, and extracted with EtOAc. The combined EtOAc portions are washed with sat'd aq NH 4 CI and brine.
The EtOAc solution is dried, filtered and concentrated to give a green-yellow colored foam, that upon purification by silica flash chromatography (7:3 hexanes/EtOAc) gives 0.879 g (1.85 mmol, 60%) of the title product: TLC (7:3 hexanes/EtOAc) Rf0.
2 1; 1
H
NMR (CDC1 3 300 MHz) d 7.39 7.00 5.46 4.61 4.13 3.80 3.62 3.20 2.13-1.53 1.42 (9H).
Preparation 47.
(Scheme I, 1-4) (S)-t--[[(1,1-Dimethylethoxy)carbonyl]amino]-3-[(2-tetrahydropyranyl)oxy]-6pyridinepropanoic acid methyl ester (C 19
H
2 8
N
2 0 6 A suspension of pre-reduced Pd/CaCO 3 (3.5 g) and 1-3 (1.15 g, 2.77 mmol) in EtOH mL) is hydrogenated (30 psi H2) for 19 h at rt. The mixture is filtered, and the filtrate is evaporated to give a yellow-colored foam, that is purified by silica flash chromatography (600:400:1 hexanes/EtOAc/iPrOH) to give 0.367 g of the title compound: TLC (1:1 hexanes/EtOAc) RfO.27; 1H NMR (CDC1 3 300 MHz) 6 8.30 7.29 7.03 (1H), 5.81 5.39 4.65 3.86 3.73 3.62 3.21 1.96-1.53 1.42 (9H).
WO 99/67230 WO 9967230PCTIUS99/1 4233 7- Preparation 48.
(Scheme 1, (S)-a-j1(1 ,1-Dimethylethoxy)carbonylj aminol-5-hydroxy-2-yridinepropanoic acid methyl ester (C 14
H
2 0
N
2 0 5 A solution of 1-4 (0.346 g, 0.91 mmol) and pyridiniumnp-toluenesulfonate (0.03 1 g, 0. 12 mmol) in EtOH (8 mL) is stirred at 55+5 'C for 20 h. The reaction mixture is cooled to rt, and concentrated in vacuo. The residue is taken up in EtOAc. The solution is washed with brine, and dried, filtered and concentrated to a pale yellow-colored oil that is purified by silica flash chromatography (500:500:1 hexaneslEtOAc/iPrOH). Evaporation of the column fractions gives 0. 132 g of the title compound: TLC 1 hexanes/EtOAc) R 0. 18; 1 HNMR (CDCI 3 300 MHz) 8 8.13 (1 7.13 (1IH), 7.03 (1 5.71 (1LH), 4.65 (1 H), 3.70 3.20 1.39 Preparation 49.
(Scheme 1, 1-6) (S)-5-(2,6-Dichlorophenyl)methoxy..c-j -1(1 ,1-dimethyletboxy)carbonylJ aminol-2pyridinepropanoic acid methyl ester (C 2 lH 2 4 C1 2
N
2 0 5 To a solution of 1-5 (0.126 g, 0.43 mmol), 2,6-dichlorobenzylalcohol (0.075 g, 0.43 mmol) and PPh 3 (0.113 g, 0.43 mmol) in dry THF (4 mL) at 0 'C under Ar is added DEAD (0.06 8 mL). The reaction mixture is permitted to warm to rt, and is stirred for 18 h. It is concentrated, and the residue is purified by silica flash chromatography (700:300:1 hexanes/EtOAc/iPrOH) to give 0. 149 g of the title compound: TLC (7:3 hexanes/EtOAc) RfO.34; I H NMR (CDCI 3 300 MHz) 8 8.31 (1 7.37 7.25 7.08 (11H), 5.81 5.29 4.65 3.70 3.24 1.63 (1 1.43 Preparation (Scheme 1, 1-7) (SX---Amino--( 2 6 -dichlorophenyl)methoxyl-2-.pyridinepropanoic acid methyl ester dihydrogen chloride salt (C I 6 H I 6 Cl 2
N
2 0 3 .2HCI).
WO 99/67230 WO 9967230PCTIUS99/1 4233 8- A solution of carbamnate 1-6 (0.546 g, 1.20 mmol) in 4 M HICl in dioxane (12 mL) is stirred at rt under Ar for 16 h. The reaction mixture is concentrated in vacuo. The residue is dissolved in H 2 0 (40 mL), and this solution is extracted with Et 2 O. The aqueous solution is frozen and lyophilized to give, as a light yellow-colored solid, 0.485 g of the title compound: 'H NMR (CD 3
SOCD
3 300 MHz) 8 8.75 8.47 7.81 (1 H), 7.57 (311), 7.48 (111), 5.35 (211), 4.49 3.67 3.42 (211).
Example 120.
(Scheme A, IS-(R I3-t(2,6-Dicbloropheny)methoxy1-6-pyridyimetbylj-2methoxy-2-oxoethyll amino] carbonyll-3-thiazo lidineca rboxylic acid (1,1dimethylethyl) ester (C 2 5
H
3 0 C1 2
N
3 0 6
S).
Example 120 was prepared as described in Scheme A from D-cysteine using the product of preparation 49 as amino acid intermediate A-4. Physical properties as follows: TLC 1 hexanes/EtOAc) Rj 0.22; 1 H NMR (CDCl 3 300 MHz) 8 8.28 7.3 8 (211), 7.28 (211), 7.09 5.29 (211), 4.90-4.74 (311), 4.40 (111), 3.67 (311), 3.38-3.22 (311), 1.61 (211), 1.40 (911).
Example 121.
(Scheme A, A-9) I -Carboxy-2-[3-[(2,6-dichlorophenyl)methoxyj-6pyridyl] ethyl] amino] carbonyll-3-thiazolidinecarboxylic acid (1 ,1-dimethylethyl) ester (C 2 4
H
2 8 C1 2
N
3 0 6
S).
Example 121 was prepared from example 120 by the procedure described in preparation 12. Physical data as follows: mp 92-94'; TLC (500:500:3 hexanes/EtOAc/HCO 2 H) Rj 0. 10; 111 NMR (CDCl 3 300 MHz) 8 8.31 (111), 8.26 (111), 7.55 (211), 7.28 (211), 7.46 (111), 7.21 (111), 5.25 (211), 4.72-4.38 (211), 4.60 (111), 4.23 (111), 3.21-3.12 (211), 3.09- 2.94 (111), 2.74 (111), 1.29 (911).
WO 99/67230 WO 9967230PCT[US99/1 4233 -159- Scheme J.
C NH 2 N J-1 ci/ CI J-2 ,NCl H 0
H
2 N 0 2HCI Scheme J teaches a general method for the preparation ofpara-acylamino derivatives of aza-phenylalanine. Thus bis-acylation of 2 -amino-5-iodopyridine J-1 gives the imide J-2, that is reacted with the organozinc, derived from a suitably protected P-iodoalanine, to provide the protected acylamino azaphenylalanine J-3. N-deprotection of J-3 gives the WO 99/67230 PCT/US99/14233 -160aminoester J-4, that is used (as exemplified by reagent A-4 of Scheme A, and by reagent of Scheme B) for the synthesis of Examples of this invention.
Preparation 51.
(Scheme J: J-2) 2-[[Bis( 2 6 -Dichlorobenzoyl)]amino]-5-iodopyridine (C 1 9
H
9 C4IN 2 0 2 To a solution of 2 -amino-5-iodopyridine J-1 (2.20 g, 10.0 mmol) and Et 3 N (2.12 mL, 15.0 mmol) in dry THF (100 mL) at rt under Ar, is added dropwise 2,6dichlorobenzoylchloride (1.60 mL, 11.0 mmol) over 45 min. The reaction mixture is stirred for 15 h. It is diluted with EtOAc (300 mL), and is washed with cold aq 1 M NaOH and brine. The solution is dried, filtered and concentrated to give a yellow-colored waxy solid, that is purified by silica flash chromatography (3:1 hexanes/EtOAc) to give 2.60 g of the title compound: TLC (7:3 hexanes/EtOAc) Rf 0.60; 1 H NMR (CDC1 3 300 MHz) 8 8.59 8.03 7.99 7.44-7.26 (6H).
Preparation 52.
(Scheme J: J-3) (S)-2-[[Bis-(2,6-dichlorobenzoyl)aminol-a-[[(1,1acid methyl ester
(C
2 8
H
2 5 Cl 4
N
3 06).
To an amberized flask containing activated Zn dust (0.865 g, 13.23 mmol) under Ar is added sequentially l-dimethylethoxy)carbonyl]-3-iodo-L-alanine methyl ester (4.36 g, 13.23 mmol), THF (13 mL) and N,N-dimethylacetamide (13 mL). The reaction mixture is purged of 02 by the bubbling Ar through the mixture for 5 min, and then is warmed to 45±5 °C for 7 h. It is cooled to rt. To this mixture is added PdC1 2 (PPh 3 2 (0.461 g) followed immediately by a degassed solution of iodide J-2 (2.60 g, 4.59 mmol) in 1:1 THF/N,N-dimethylacetamide (18 mL). The reaction mixture is stirred at 45±5 °C under Ar for 13 h. It is cooled to 0 OC and quenched with satd aq NH 4 Cl (150 mL). The mixture is extracted with EtOAc. The combined EtOAc extracts are washed with brine, dried, filtered and concentrated to a green-yellow-colored paste, that is purified by silica WO 99/67230PCIS9143 PCTIUS99/14233 -161flash chromatography (700:300:1 hexanes/EtOAc/iPrOH) to give 1.43 g of the title compound: TLC (7:3 hexanes/EtOAc) RfO0.29: 1 H NMR (CDCI 3 300 MHz) 8 8.13 (1 H), 7 (1 7.46 0IH), 7.26 4.80 (1 4.50 3.67 3.05 1.46 (9H).
Preparation 53.
(Scheme J: J-4) (S-a-Amino-2-j [bis-( 2 6 -dicblorobenzoyl)laminol-s...pyridinepropanoic acid methyl ester dihydrochioride salt (C 2 3
H
1 7 C1 4
N
3
O
4
-HCI).
A solution of J-3 (0.69 g, 1.08 mmol) in 4 M HCl in dioxane (15 mL) is stirred under Ar for 20 h. The reaction mixture is concentrated in vacuo, diluted with and extracted with Et 2 O. The aqueous solution is frozen and lyophilized to give, as a pale yellowcolored solid, 0.627 g of the title compound: 1 H NMR (CD 3
SOCD
3 300 MHz) 6 8.80 8.27 (1 7.82 (1IH), 7.67-7.26 4.25 (1 3.52 3.16 3.04 (1 H); MIS (ESI+) m/z 541.7.
Example 122.
(Scheme A, IBis-(2,6-dichlorobenzoyl) jaminoj-5-pyridylj methyll-2methoxy-2-oxoethyll aminolicarbonyll-3-thiazolidinecarboxylic acid (1,1dimethylethyl) ester (C 2 5
H
3 0 Cl 2
N
3 0 6
S).
Example 122 was prepared as described in Scheme A from D-cysteine using the product of preparation 52 as amino acid intermediate A-4. Physical properties as follows: TLC (1:1 hexanes/EtOAc) Rf 0.22; 1 H NMR (CDC1 3 300 MHz) 8 8.28 7.38 7.28 (2H), 7.09 5.29 4.90-4.74 4.40 (1 3.67 3.38-3.22 1.61 (2H), 1.40 (9H).
Example 123.
(Scheme A, A-9) pyridylil ethyljI amino Ica rb onvl-3-thiazo lid ineca rboxy lic acid (1 ,1-dimethylethyl) ester (C 31 I H-.
8 C1 4
N
4 0 7
S).
WO 99/67230 PCT/US99/14233 -162- Example 123 was prepared from example 122 by the procedure described in preparation 12. Physical data as follows: mp 158-1600; TLC (50:50:2 hexanes/EtOAc/HCO 2 H) Rf 0.18; 1 H NMR (CD 3
SOCD
3 300 MHz) 5 8.42 8.22 7.75 7.69-7.17 7.53 4.59 4.40 4.19 3.19-3.02 2.84 2.56 (1H), 1.34 (9H).
Scheme K.
R3
OH
N 0 K-3 Rs
H
2 N\ °0 K-2
H
0 N R OH K-4 R3Y O
R
Scheme K teaches a general method for the preparation of oxazolidinecarboxylic acid Examples K-3 and K-4, where R3, R 5 and Y are identical to the definitions of Scheme B.
Thus coupling of oxazolidinecarboxylic acid K-1 and aminoester K-2 (as exemplified by the reaction of reagents A-3 and A-4 of Scheme A, and B-4 and B-5 of Scheme B) provides Examples K-3, that are hydrolyzed to Examples K-4 of this invention.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -163- Example 124.
(Scheme K: K-3. where R 3 is (phenyl)methyl. R, is 4 2 6 -dichlorobenzoyl)amino]phenyl, Y is CO, and the stereochemistry is R 1 4 -I(2,6-Dicblorobenzovl)aminol pbenyl~methylJ-2-methoxy-2oxoethyl] amino Ica rbonylj-3-oxazolidin eca rboxy lic acid 3-phenylmethyl ester
(C
29
H
27 C1,N 3 0 7 Example 124 was prepared as by coupling commercially available (.S)-3,4-oxazolidinedicarboxylic acid 3-(phenylmethyl) ester to amino acid K-2 (Scheme K, where R, is 4-[2,6dichlorobenzoyl)amino]phenyl stereochemnistry is under the conditions described by preparation Physical properties as follows: TLC (3 :2 Heptane/EtOAc Rf 0. 17; UV (MeOH) ma,225 (e 12600, sh), 251 (17900); 3 C NMR (d 6 -dimethylsulfoxide) 8 188.75.
171.96, 169.86, 162.40, 162.30, 152.99, 137.59, 137.47, 136.87, 136.81, 133.23, 131.79, 131.65, 130.08, 128.80, 128.67, 128.28, 127.76. 119.84, 119.75, 79.82, 66.77. 57.85, 53.85, 52.47, 36.62 (23 lines expected; 26 lines observed); MS (FAB) m/z 602. 600, 558, 556, 531, 466, 371, 351, 349, 280, 278, 175. 173: MS, (FAB) m/z 600.1312 (calcd 600.1304; Anal. C, 57.75; H, 4.75; N, 6.80; Cl, 11.86 (calcd for 0.42% H,O: C 57.77, H 4.56, N 6.97, CI 11.76).
Example 125.
(Scheme K: K-4 where R 3 is (phenyl)methyl. R, is 4-II(2,6-dichlorobenzovl)amino]phenyl, Y is CO, and the stereochemistry is [I l-Carboxy-2-[4-I(2,6-dichlorobenzoyi)aminolphenylI ethyl] arnino]carbonyl]-3-oxazolidinecarboxylic acid 3-phenvlmethyl ester (C 2 8
H
2 5 ClN 3 0 7 Example 125 was prepared from example 124 by the procedure described in preparation 12.
Physical properties as follows: TLC (950:50:1 CH,Cl,/MeOH/HCO 2 H) Rf=0.34; 'H NMR (d 6 -dimethylsulfoxide) 8 10.65 (1IH). 8.31 (I1H), 7.61-7.42 7.40-7.20 7.15 5.19-4.8 5 4.90 (1lH), 4.76 (1iH), 4.43 (1IH), 4.3 6 (1 4.11 (1lH), 3.65 (I1H), 3.04 (1 2.87 (1IH); MS (FAB) m/z 588. 586 544, 542, 532. 391, -337, 33 5, 327, 269,267, 161. 147.133, 129, 117,115, 103, 101.91: MS (FAB) 586.1132 (calcd 586.1147).
WO 99/67230 WO 9967230PCTIUS99/1 4233 -164- Example 126.
(Scheme K: K-3 where is benzyl, R, is 4-[(2,6-dichlorobenzoyl)aminolphenyl, Y is CO, and the stereochemistry is [11-I14-I(2,6-Dichlorobenzoyl)aminol phenvilmethyll-2-methoxy-2oxoethyll amino] carbonylj-3-oxazolidinecarboxylic acid 3-phenylmethyl ester
(C
29
H
2 7C1 2
N
3 0 7 Example 126 was prepared as described in Scheme K from commercially available 3, 4 -oxazolidinedicarboxylic acid 3-(phenylmethyl) ester. Physical properties as follows: TLC (3:2 heptane/EtOAc) Rf 0. 19; UV (MeOH) 225 (e 12400, sh), 252 (17700), 284 (2960, sh); "C NMR (CDCl 3 6 171.23, 169.14, 162.43. 154.26, 136.47, 135.93, 135.65, 132.37, 132.28, 130.91, 130.00, 128.62, 128.39, 128.1 1, 120.30, 79.66, 67.94, 58.38, 53.14, 52.49, 37.19 (23 lines expected; 21 lines observed); MIS (FAB) m/lz 602, 600, 558, 556, 466, 351, 349. 280, 278, 175, 173; MIS (FAB) m/z 600.1299 (calcd for 600.1304); Anal. C 57.69, H 4.90, N 6.71, CII 11.49 (calcd for 0.35% H,O: C 57.81, H 4.56, N 6.97, CI 11.77).
Example 127.
(Scheme K: K-4 where R 3 is benzyl, R 5 is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, Y is
CO
2 and the stereochemnistry is dichlorobenzoyl)aminol phenylj ethyl] aminol-carbonylj-3-oxazolidinecarboxylic acid 3-.(phenylmethyl) ester (C 2
,H
25 Cl2, 3 0 7 Example 127 was prepared from example 126 by the procedure described in preparation 12.
Physical properties as follows: TLC (950:50:1 CI- 2 C,/MeOHII-CO,H) Rj 0.3 1; 'H NMR (d 4 -methanol) 8 7.60 (2H),7.49-7.23 7.21 5.23-4.97 (211), 4.95 (1 H), 4.88 (1 4.70 (1IH). 4.36 (1lH). 4.14 (1 3.85-3.73 (1lH), 3.23 (1 3.00 (1 MIS (FAB) m/z 588, 586. 544. 542, 532, 391, 3 71, 337, 3 35, 245. 177, 173.) 149. 123, 105, 103, 91; MS (FAB) m/z 586.1163 (calcd 586.1147).
WO 99/67230 WO 9967230PCTIUS99/1 4233 -165- Scheme L.
RL-2 'IRL-I
RL.
2
RL-I
0 N L-2 -0 OH
H
2 N OL-3
RL-
2 Y RL-1 O N 0 L-4
R
RL-
2
RL-I
O N 0 o Ho HN
OH
Scheme L teaches a general method for the preparation of N-alkylaryl azetidinedicarboxylic acid Examples L-4 and L-5, where R 1 1 s Cl.
6 alkyl, RL.
2 is C 6 1 0 aryl and R, is defined as in Scheme B. The N-phenylethyl-2.4-azetidinedicarboxylic acid dimethyl ester stereoisomers of general structure Li were prepared as described (Hoshino. Hiraoka, J.; Hata. Sawada, Yamamoto. Y. J. Chem. Soc. Perkin Trans. 1 1995, 693-697) and separated by silica flash chromatography. Thus partial saponification of diester L-I gives WO 99/67230 WO 9967230PCT/US99/I 4233 -166the half-acid L-2. that is coupled with reagent L-3 (as exemplified by the use of reagents A-3 and A-4 of Scheme A. and B-4 and B-5 of Scheme B) to provide Examples L-4, that are then hydrolyzed to Examples L-5 of this invention.
Preparation 54.
(Scheme L: L-I where is methyl, RL.
2 is phenyl and stereochemistry is [2S- [1 12S-[l1(R*),2cx,4p1 1-1-(1 -Phenylethyl)-2,4-azetidinedicarboxylic acid dimethyl ester [168647-92-5] (Cl 5
H
19 N0 4 Physical properties as follows: TLC (4:1 Hexanes/EtOAc) Rf 0.42; 3 C NMR (CDCl 3 8 173.61, 142.41. 128.31. 127.39. 127.35, 60.95. 60.67, 51.64, 24.96, 21.72.
Preparation (Scheme L: L-1 where RL.I is methyl, RL- 2 is phenyl and stereochemnistry is [2R- [1 12R-I 1(S*),2a,4p5j] -Phenylethyl)-2,4-azetidinedica rboxylic acid dimethyl ester
(C
15
HI
9 N0 4 Physical properties as follows: TLC (4:1 Hexanes/EtOAc) R 0.3 1; 3 C NMR (CDCl 3 8 172.97, 141.21, 128.44, 128.07, 127.68, 61.54, 60.69, 51.61. 24.91, 19.55.
Preparation 56.
(Scheme L: L-1 where RL-I is methyl, RL- 2 is phenyl and stereochemistry is [I1(S)-cis]).
[1 (S)-cisj-1 -Phenylethyl)-2,4-azetidinedicarboxylic acid diniethyl ester [168753-32- 0] (C 15
H
19 N0 4 Physical properties as follows: TLC (4:1 Hexanes/EtOAc) Rf= 0.21; 3 C NMR (CDC1 3 6 172.58, 172.08, 140.84, 128.22. 128.15, 127.65, 66.32. 60.10. 59.65, 52.06, 51.59, 24.26, 19.91.
Preparation 57.
(Scheme L: L-I where RL.I is methyl, RL.
2 is phenyl and stereochemistry is [2R- [1 [2R-[1 (R*),2a,431 I-(1-Pbenylethyl)-2,4-azetidinedicarboxylic acid dimethyl ester
(C
15
H
19 N0 4 WO 99/67230 PCT/US99/14233 -167- Physical properties as follows: TLC (8:2 hexanes/EtOAc) Rf= 0.42; "C NMR (CDC13) 173.61. 142.41, 128.31, 127.39, 127.35, 60.95, 60.67, 51.64, 24.96. 21.72; MS (+ESI) m/z 278.3.
Preparation 58.
(Scheme L: L-1 where RL., is methyl, RL.2 is phenyl and stereochemistry is [2S- [2S-I1(S*),2ca,4p]]-l-(l-Phenylethyl)-2,4-azetidinedicarboxylic acid dimethyl ester (CIsH,,NO 4 Physical properties as follows: TLC (8:2 hexanes/EtOAc) Rf= 0.31; "C NMR (CDCI 3 172.97. 141.21, 128.44, 128.07, 127.68, 61.54, 60.69, 51.61, 24.91. 19.55: MS (+ESI) m/z 278.3.
Preparation 59.
(Scheme L: L-1 where RL. is methyl, RL2 is phenyl and stereochemistry is I1(R)-cisl-l-(1-Phenylethyl)-2,4-azetidinedicarboxylic acid dimethyl ester
(CIH,,NO
4 Physical properties as follows: TLC (8:2 hexanes/EtOAc) Rf= 0.21; "C NMR (CDCl 3 172.58, 172.08, 140.84, 128.22, 128.15, 127.65, 66.32, 60.10, 59.65, 52.06, 51.59, 24.26, 19.91; MS (+ESI) m/z 278.3.
Preparation (Scheme L: L-2 where RL.I is methyl, RL.2 is phenyl and stereochemistry is [2S- [2S-[i(R*),2a,41]]-1-(1-Phenylethyl)-2,4-azetidinedicarboxylic acid monomethyl ester
(C,
4
H,
7
NO
4 A mixture of L-1 (Scheme L, where RL.- is methyl, RL.
2 is phenyl and stereochemistry is the product of preparation 54) (7.95 g; 28.7 mmol) and LiOH mmol) in 1:1 MeOH/HO (240 mL) is stirred at rt for 42 h. The reaction mixture is adjusted to pH 5 with HOAc, and is concentrated. The resulting concentrate is diluted with brine and extracted repeatedly with CHCI 3 The combined CHCI, extracts are dried, filtered and concentrated to give a yellow foam (6.61 that is purified by preparative C 18 reverse phase chromatography to give the title compound as a crystalline solid: mp WO 99/67230 WO 9967230PCTJUS99/1 4233 -168- 112-1 13 TLC (650:350:1 hexanes/EtOAc/HCO,H) Rf= 0.17; MIS (FAB) /z 527, 264, 248, 218, 204. 192. 186. 177, 160, 114, 105; Anal. C 64.04, H 6.57, N 5.37 (calcd C 63.8 7, H 6.5 1. N 5.3 2).
Preparation 61 and Example 128.
(Scheme L: L-4 where RL.I is methyl, RL.
2 is phenyl, R, is 4-[(2,6-dichlorobenzoyl)amnino]phenyl and stereochemistry is [2S- [1(R I2S-I1(R*),2ct,43(R*)I]-4-ItI1-[ [4-[(2,6-Dichlorobenzoyl)aminolphenyllinethylj-2methoxy-2-oxoethyll amino Icaronyll.-I -(Il-p henylethyl)-2-azetidinecarb oxyl ic acid methyl ester (C 3 1
H
3 ,C1 2
N
3 0 6 A mixture of L-2 (Scheme L, where RL-l is methyl, RL- 2 is phenyl and stereochemistry is the product of preparation 60) (0.62 g, 2.4 mmol), L-3 (Scheme L where R, is 4 2 6 -dichlorobenzoyl)amino)phenyl and stereochemistry is (0.95 g, 2.4 mmole), and BOP-CI (0.68 g; 2.7 mmol) in CH2Cl.,(1 0 mL) is treated with (i-Pr) 2 NEt (1.7 mL, 9.8 mmol). The reaction mixture is stirred at r1 under N 2 for 19 h. It is diluted with half-saturated NaHCO 3 and extracted with CHCI 2 The CH 2
CI
2 extracts are dried, filtered and concentrated to give a beige-colored foam (1.40 that is purified by silica flash chromatography to give the title compound: TLC 1 Hexanes/EtOAc) Rj 0.23; "C NMR (CDCl 3 6 173.25, 172.73, 171.63, 162.57.141.86, 136.76, 136.01, 132.44, 132.39, 130.90, 130.00, 128.57, 128.12, 127.55, 127.08. 120.37, 63.25, 60.06, 59.17, 52.41, 52.12, 51.47, 37.65, 26.06, 21.25; MIS (FAB) m/z 612.1658: Anal. C 59.66, H 5.14, N 6.64 (calcd C 60.79, H 5. 10, N, 6.86).
Preparation 62 and Example 129.
(Scheme L: L-5 where RL-1 is methyl, RL.
2 is phenyl, R, is 4-[(2,6-dichlorobenzoyl)axnino]phenyl and stereochemistry is I2S-I1(R*),2x,4p3(R*)I 1-4-111-Carboxy-2-[4-1(2,6-dichlorobenzoyI)aminolphenyJ..
ethyl]l-amino] ca rbonylj-I1 -p henylethyl)-2-azetidinecarboxylic acid dilithiumn salt
(C
2 qH 25 C1 2 Li2_N 3
O
6 A mixture of L-4 (Scheme L, where RL.
1 is methyl, RL.
2 is phenyl, R, is 4-[2,6dichlorobenzovl)amino]phenyl and stereochemnistry is the product of preparation 61) (0.684 g, 1.12 mmol) and LiOH*H,O (0.25 g, 6.0 mmol) is dissolved by WO 99/67230 WO 9967230PCTIUS99/1 4233 -169warming in MeOH (10 mL). This solution is diluted with 1: 1 H,0/THF (20 mL), and the reaction mixture is stirred at rt for 22 h. The mixture is adjusted with 1 N HCl to pH 6. The solution is concentrated to give a white solid, that is purified by preparative Cl 18 reverse phase chromatography (MeCN/H,O gradient). Evaporation of the column fractions gives a white solid, that is dissolved in warm H 2 0. The solution is frozen and lyophilized to give, as a white solid, the title compound: mp 270 TLC (850:150:1 CHCI 1 /MeOH/HCO 2
H)
Rf=r 0.21-0.36; 3 C NMR (CD.3OD) 8 178.05, 174.59, 174.01. 162.20. 142.33, 135.09, 134.91, 133.04, 130.54. 129.24. 128.61, 126.46, 126.41,126.17,.124.94, 118.33,82.70, 61.70, 60.50. 53.30, 36.23. 25.85. 18.65; MS (FAB) m/z 584.1350: Anal. C 55.17, H 5.01, N 6.63, Cli 11.16 (calcd for 7.52% H,O: C 54.02, H 4.75, N 6.52. Cl. 11.2 1).
Example 130.
(Scheme L: L-4 where is methyl, RL.
2 is phenyl, R, is dichlorobenzoyl)amino]phenyl and stereochemnistry is I(S* ),2cx,413(S* I2R-II(S*),2a,4(S*)I-4-III 1 4
-I(
2 ,6-Dichlorobenzoyl)aminojpbenvlmethyl j-2methoxy-2-oxoethyll amino] carbonylj-1 -phenylethyl)-2-azetidinecarboxylic acid methyl ester (C 3
,H
31
C,N
3 0 6 Example 130 was prepared as described in Scheme L using the product of preparation as intermediate L-1. Physical properties as follows: TLC EtOAc/Hexanes) Rf 0.43; [l] 2 sD +58 (c 0.9 1. MeOH); 'C NMR (CDC1 3 8 173.24. 172.15. 172.05, 162.5 8, 140.75, 136.45, 136.01. 132.89, 132.38, 130.91, 129.60, 128.67. 128.39, 128.11, 127.97, 120.39, 63.54, 60.19, 59.54. 53.15. 52.16, 51.91, 37.86, 26.12. 18.37, MS (El) m/z 613, 611, 598, 596, 554. 552, 527. 525, 508, 506, 450, 448, 351. 349. 218, 191, 175, 173, 160, 131, 114, 105; Anal. C 60.68. H 5.18, N 6.67, CI 1.24 (calcd for C 60.79. H 5. 10, N 6.86, Cl 11.58).
Example 13 1.
(Scheme L: L-5 where RL.1 is methyl, RL- 2 is phenyl, R, is 4-[(2.6-dichlorobenzoyl)aminoiphenyl and stereochemnistry is I(S* [2R-I 1(S*),2cL,4f3(S*)] 1-4-11 [1-Carboxy-2-[4-1(2,6-dichlorobenzoyi)aminolphenvlethylI-aminolcarbonyll...1(I -phenylethyl)-2-azetidinecarboxylic acid, dilithium salt (C, 9 H,,CI,Li.N.1O 6 WO 99/67230 PCT/US99/14233 -170- Example 131 was prepared from Example 130 by the procedure described in preparation 62. Physical properties as follows: [ca] 25D +84 (c 0.95, MeOH); 3 C NMR (CD30D) 8 179.18. 176.55, 174.13, 163.58, 141.88. 136.32, 136.29, 134.96, 131.90. 130.86. 129.47, 128.49. 128.07,127.87, 127.16, 119.74, 63.70, 63.49, 59.77, 56.27, 38.24, 26.25. 18.70; MS (FAB) m/z 598, 596, 592, 590, 552, 550, 546. 544, 161; Anal. C 55.20, H 5.52, N, 6.59 (calcd for 10.98% H 2 0: C 52.00, H 4.99, N 6.27).
Example 132.
(Scheme L: L-4 where RLI is methyl, RL.
2 is phenyl, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is a single diastereomer having a cis relative configuration but unknown absolute configuration at C-2 and C-4 of the azetidine.) [1(S),2a,4ao(S)]-4-[[[1-[[4-[(2,6-Dichlorobenzoyl)amino]phenyl]methyll-2-methoxy-2oxoethyl]amino]carbonyl]-l-(1-phenylethy)-2-azetidinecarboxylic acid 2-methyl ester (C 31
H
3
,CIN
3 06).
Example 132 was prepared as described in Scheme L using the product of preparation 56 as intermediate L-1. Physical properties as follows: TLC (9:1 CHCl 3 /acetone) Rf= 0.29; 3 'C NMR (CDC1 3 172.70, 172.29, 171.65, 162.49, 140.97, 136.71, 136.12, 132.47, 132.39. 130.81, 130.22, 128.35, 128.07, 127.82, 120.17, 66.61, 61.52, 59.99, 52.38, 52.24, 51.79. 37.73. 25.16, 20.09.
Example 133.
(Scheme L: L-5 where RL,, is methyl, RL.
2 is phenyl, R 5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is [1 a single diastereomer having a cis relative configuration but unknown absolute configuration at C-2 and C-4 of the azetidine.) 1 (S),2a,4oa(S)-4-[[[1-Carboxy-2-[4-[(2,6-dichlorobenzoyl)amino]phenyl]ethyl]aminol-carbonyl-l-(l-phenylethyl)-2-azetidinecarboxylic acid (C2 9
H
27 C1N 3 0 6 Example 133 was prepared from Example 132 by the procedure described in preparation 62. Physical properties as follows: [c]2 D +20 (c 0.88, MeOH); "C NMR (CD 3 OD) 6 173.70. 173.05. 172.61, 163.69, 140.09, 136.93, 136.20. 133.18, 131.89. 130.89, 129.95, 128.09. 127.89.127.88. 127.65, 120.03, 66.07, 61.29. 59.86. 52.70. 36.98. 25.03. 18.81; WO 99/67230 WO 9967230PCTIUS99/1 4233 -171- MIS (FAB) m/lz 662. 660. 586. 584, 539, 482, 480,.436. 434. 204. 175, 173. 160, 133, 109, 105; Anal. C 57.90, H 5.01, N 6.93 (calcd for 4.98% H,O: C 56.63, H 4.98, N 6.83).
Example 134.
(Scheme L: L-4 where RL.1 is methyl, RL.
2 is phenyl, R 5 is 4-[2,6-dichlorobenzoyl)aminolphenyl and stereochemnistry is [2R-[1 [2R-I 1(R*),2ct,43(S*)1 1-4-F I-F 14-I(2,6-Dichlorobenzoyl)aminolphenyllmetbyll-2methoxy-2-oxoethyll amino] carbonyll -1 -phenylethyl)-2-azetidinecarboxylic acid methyl ester (C! 3
,H
3 ,C1,N3O 6 Example 134 was prepared as described in Scheme L using the product of preparation 57 as intermediate L-1. Physical properties as follows: TLC 1: 1 EtOAc/Hexanes) Rf 0.30.- 3 CNMR (CDC1 3 6 173.78. 172.76, 172.10, 162.47, 141.81, 136.70, 135.96, 132.43, 132.38, 130.88, 129.79. 128.55, 128.10, 127.58, 127.00. 120.58, 63.32, 60.06, 59.27, 52.70, 52.50, 51.44, 37.08. 25.76, 21.52; MS (+ESI. 200:1 MeOH/HCO,H solution) ,n/z 613.8, 611.8.
Example 135.
(Scheme L: L-5 where RLI1 is methyl, RL.
2 is phenyl, R 5 is 4-[2,6-dichlorobenzoyl)aminoiphenyl and stereochemnistry is I (R*),2L,413(S* )II) I2R[I1(R*),2c-,4p(S*)1 1-4-F[I-Carboxy-2-14-(2,6-dichlorobenzoyl)aminolphenylJethyl]l-amino] carbonyl 1-1 -p henylethyl)-2-azetidin eca rboxylic acid dilithium salt
(C
29
H
25 C1,Li 2
N
3
O
6 Example 135 was prepared from Example 134 by the procedure described in preparation 62. Physical properties as follows: [a] 2 1D +76 (c 0.89, MeGH); 3 C NMR (CD 3 OD) 6 179.44, 176.71, 175.90. 163.55, 143.81, 136.54, 136.26. 134.47, 131.90, 120.81, 129.70, 127.83, 127.34, 126.333, 119.88. 63.06, 62.11, 58.71, 55.46. 37.17. 26.76, 20.56; MIS (FAB) m/z 598, 596, 592. 590. 552, 550, 546, 544,237. 105; Anal. C 55.12, H 5.24, N 6.59, CI 10.56 (calcd for 6.2 1% H,O: C 54.78, H 4.66, N 6.6 1, Cli 11.15).
Example 136.
(Scheme L: L-4 where is methyl, RL.
2 is phenyl, R, is 4-[(2,6-dichlorobenzoyl)aminolphenyl and stereochemistry is I WO 99/67230 WO 9967230PCTIUS99/1 4233 -172- 12S-I 1 (S*),2t,403(R*)j 1-41 111-I14-1(2,6-Dichlorobenzoyl)aminoI phenvllmethyll-2methoxy-2-oxoethylj amino] carbonyll- 1 -phenylethyl)-2-azetidinecarboxylic acid methyl ester (C.
31
H
31
C,N
3 0 6 Example 136 was prepared as described in Scheme L using the product of preparation 58 as intermediate L-1. Physical properties as follows: TLC (85:15 CHCI 3 /acetone) Rj 0.54; 1 3 CNMR (CDC1 3 5 173.44, 172.62, 171.72, 162.79, 140.46, 136.69, 136.34. 1-3 3.12, 132.79, 131.29, 130.40, 129.08, 128.52, 128.45, 128.23, 120.57, 64.15, 60.51. 60.09, 53.11, 52.59, 52.19, 37.73, 26.70, 18.75; Anal. C 60.70, H 5.39, N 6.62 (calcd C 60.79, H 10, N 6.86).
Example 137.
(Scheme L: L-5 where RL-I is methyl, RL- 2 is phenyl, R, is 4-[(2,6-dichlorobenzoyl)aminoiphenyl and stereochemnistry is 2S-[ 1 1(S*),2ct,413(R*)I 1-4-I II1-Carboxy-2-14-I(2,6-dichlorobenzoyl)aminoj phenyl]ethyl]l-aminolIcarbonyl 1 -phenylethyl)-2-azetidin ecarboxylic acid (C 29 ,H,,C1 2
N
3 0 6 Example 137 was prepared from Example 136 by the procedure described in preparation 62. Physical properties as follows: [CC] 25 D +2 (c 1.00, MeOH); 3 C NMR (CDOD) 8 172.31, 169.43, 167.37, 163.72, 161.39, 160.93, 136.98, 136.11, 134.40, 133.20, 131.84, 130.93, 129.42, 129.39, 128.60, 127.89. 120.11. 62.68, 61.41, 53.51, 36.35. 25.00, 16.17; MS (FAB) m/z 586, 584, 482, 480, 204, 175. 173, 106, 105; Anal. C 53.07. H 4.52, N 6.15, Cl 10.46 (calcd for 0.80 equiv TFA and 2.13% H 2 0: C 53.20, H 4.30, N 6.08, Cl 10.26).
Example 138.
(Scheme L: L-4 where RL-I is methyl, 2 is phenyl, R, is 4-[2,6-dichlorobenzoyl)aminolphenyl and stereochemnistry is [1 (R),2cc.4cc(S)J, a single diastereomer having a cis relative configuration but unknown absolute configuration at C-2 and C-4 of the azetidine.) 11 (R),2ot,4ct(S)j -4-1[I1l-114-I(2,6-Dichloro benzoyl)aminol phenyljminethyl I-2-methoxy-2oxoethyll amino]lcarbonyll -1 -phenylethyl)-2-azetidinecarboxylic acid 2-methyl ester 1
H
31 C1,N 3 0 6 WO 99/67230 PCT/US99/14233 -173- Example 138 was prepared as described in Scheme L using the product of preparation 59 as intermediate L-1. Physical properties as follows: TLC Rf= 0.47 (85:15 CHC1 3 /acetone); "C NMR (CDCI 3 6 173.42, 172.40. 171.74. 162.46, 140.80, 136.76, 136.03, 132.64, 132.35, 130.80, 129.82, 128.38, 128.04, 127.85, 127.69, 120.39, 66.11, 61.50, 59.78, 52.83, 52.42, 51.72, 37.12, 25.03, 20.20; MS (+ESI, 200:1 MeOH/HCO,H solution) m/z 614.2. 612.2; Anal. C 60.66, H 5.18. N 6.80. Cl 11.42 (calcd C 60.79, H 5.10, N 6.86, Cl 11.58).
Example 139.
(Scheme L: L-5 where RL. is methyl, RL.
2 is phenyl, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is a single diastereomer having a cis relative configuration but unknown absolute configuration at C-2 and C-4 of the azetidine) [1 (R),2a,4o(S)]-4-[[I1-Carboxy-2-[4-I(2,6-dichlorobenzoyl)aminolphenyllethyl]amino]-carbonyl]-l-(1-phenylethyl)-2-azetidinecarboxylic acid (C2 9 H,2Cl 2
N
3 0 6 Example 139 was prepared from Example 138 by the procedure described in preparation 62. Physical properties as follows: [a] 25 D -30 (c 0.90. MeOH); 3 C NMR (CD30D) 6 172.71, 172.44, 171.81, 163.71, 138.03, 136.99, 136.15, 133.31, 131.89, 130.89, 129.45, 128.33, 128.29,128.02, 127.86, 120.23, 65.62, 61.51. 59.60, 53.47, 36.04, 24.91, 17.68; MS (FAB) n/z 586. 584, 371, 298, 204, 177, 175. 173. 133. 105, 100; Anal. C 55.69, H 4.49, N 6.55, Cl 11.81 (calcd for 0.38 equiv TFA and 2.06% HO: C 55.80, H 4.54, N 6.56, Cl 11.07).
WO 99/67230 WO 9967230PCTIUS99/1 4233 -174- Scheme M.
RM.2 YRWI -0 O-M-1
H
O N 0 -0o 0 M-2
R
3 M-3 -0 0
R
3 M-4 -0 OH
H
2 N OlM- 0
R
3 O N' 0 -0 HN, 0 M-6
R
R
3 O0 HO HN 'OHM-7
R
WO 99/67230 PCT/US99/14233 -175- Scheme M teaches a general method for the preparation of N-acyl azetidinedicarboxylic acid Examples M-6 and M-7, where RM., is alkyl, R.
2 is C 6 10 aryl, and R, and Y are defined as in Scheme B. Thus removal of the N-alkylaryl substituents Rm., and RM.
2 of M-1 gives aminodiester M-2, that is acylated to provide M-3. Partial saponification of diester M-3 gives half-acid M-4, that is coupled with reagent M-5 (as exemplified by the use of reagents A-3 and A-4 of Scheme A, and B-4 and B-5 of Scheme B) to provide Examples M-6, that are then hydrolyzed to Examples M-7 of this invention.
Preparation 63.
(Scheme M: M-2 where stereochemistry is (2R-trans)) 2 R-trans)-2,4-Azetidinedicarboxylic acid dimethyl diester (C,H,,N0 4 To an N,-purged solution of freshly chromatographed Preparation M-1 (Scheme M where RM., is methyl, RM- 2 is phenyl and stereochemistry is prepared as described by preparation 57) (656 mg, 2.37 mmol) in MeOH (20 mL) is added Pd(OH) 2 /C (120 mg), and this mixture is hydrogenated for 19 h under an H. atmosphere (approximately 42 psi pressure). The mixture is filtered and concentrated to give the title compound as a colorless oil: TLC (4:1 Hexanes/EtOAc) Rf= 0.04; 'H NMR (CDC1 3 8 4.33 3.77 3.38 2.71 MS (+ESI) m/z 174.2.
Preparation 64.
(Scheme M: M-3 where R 3 is ethyl, Y is and the stereochemistry is (2R-trans)) (2R-trans)-1, 2 ,4-Azetidinetricarboxylic acid 1-ethyl-2,4-dimethyl triester (CoH1sNO,).
To a mixture of freshly prepared amine M-2 (Scheme M where stereochemistry is (2Rtrans)) (14 mmol) in CH 2 Cl 2 (20 mL) under N, at 0 OC is added Et 3 N (3.0 mL, 22 mmol), followed by the dropwise addition of CICOEt (1.5 mL, 18 mmol). After 22 h the reaction is quenched with saturated NaHCO,, diluted with H,O. and extracted with EtOAc. The combined organic extracts are dried, filtered and concentrated to give 2.79 g of the carbamate, that is purified by silica flash chromatography: TLC (4:1 Hexanes/EtOAc) Rf= 0.17; [ca] 2 5 +183 (c 0.83, MeOH); 1 H NMR (CDCI,) 8 4.77 4.15 4.10 (1H), 3.80 2.58 1.23 NMR (CD,OD) 8 173.11, 173.05. 157.47. 63.30, 61.00, 60.18. 53.41. 26.43, 15.27; MS (El) i/z 245. 186, 172, 142. 114; Anal. C 48.90. H 6.21, N 5.73 (calcd C 48.98, H 6.16, N 5.71).
WO 99/67230 PCT/US99/14233 -176- Preparation (Scheme M: M-4 where R 3 is ethyl, Y is CO 2 and the stereochemistry is (2R-trans)) (2R-trans)-1,2,4-Azetidinetricarboxylic acid I-ethyl-2-methyl diester (C 9
H,
3 NOs).
A mixture of M-3 (Scheme M, where R 3 is ethyl, Y is and the stereochemistry is (2R-trans)) (1.68 g, 6.85 mmol) and LiOH (7.00 mmol) in 1:1 MeOH/H0O) (40 mL) is stirred at rt for 45 h, and then is concentrated. The residue is dissolved in half-saturated NaHCO 3 and the solution is extracted with EtO. The Et,O solution is discarded. The aqueous solution is adjusted with concentrated HCI to pH 4. and is concentrated to a yellow solid. This solid is triturated with CHC1 3 The CHC1, solution is filtered and concentrated to give, as a light brown oil, the title compound: TLC (600:400:1 Hexanes/Acetone/HCOH) Rf= 0.21-0.45; 'H NMR (CDC 3 8 4.87 4.77-4.68 (1H), 4.64-4.50 4.18-3.97 3.76 2.52-2.33 1.26-1.13 MS (-ESI, MeOH solution) m/z 230.1. This material is used without purification.
Preparation 66 and Example 140.
(Scheme M: M-6 where R 3 is ethyl, R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl, Y is
CO
2 and the stereochemistry is [2R-(2at,4P(S*)]-4-[[[l-[[4-l(2,6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyljamino]carbonyl]-l,2-azetidinedicarboxylic acid 1-ethyl-2-methyl diester
(C
26
H
2 7
CIN
3 0 8 To a mixture of Preparation M-4 (1.08 g, 4.67 mmol) and HOBt+H,O (0.63 g, 4.7 mmol) in CH,CI, (10 mL) at 0 OC is added a solution of EDC (1.04 g, 5.42 mmol) in CH 2 C1l mL). This mixture is stirred at 0 C for 30 min. It is then treated with M-5 (Scheme M where R 5 is 4-[(2,6-dichlorobenzoyl)amino)phenyl and stereochemistry is (1.88 g, 4.66 mmol) and N-methylmorpholine (0.52 mL, 4.7 mmol). The resulting solution was stirred at 0°C for 2 h and at rt for 2 h. The reaction mixture is diluted with 10% KHSO 4 and extracted with CH.CIl. The combined CH,CI, extracts are washed with satd NaHCO 3 and brine, and are combined, dried, filtered and concentrated to give a yellow foam (2.58 that is purified by flash chromatography to give, as a white solid, the title compound: mp 97-99 0 C; TLC (9:1 CHCl,/acetone) Rf= 0.30; [a] 2 o D +79 (c 1.02. MeOH); UV (MeOH) Xma 225 (E 12200: sh), 251 (17500); 3 C NMR (CDOD) 8 171.50, 171.30, WO 99/67230 PCT/US99/14233 -177- 171.08. 163.67, 156.23, 155.96, 136.92. 136.19, 133.21, 133.03, 131.87. 130.93, 129.64, 129.44, 127.92, 119.97, 61.61, 60.56, 59.82. 59.11. 58.41, 53.45, 51.58. 51.52. 36.57, 36.29. 24.91. 13.43 (22 lines expected; 28 lines observed); MS (FAB) m/z 580.1260 (calcd for 580.1253); Anal. C 51.95, H 4.56, N 6.89, Cl 15.18 (calcd for 0.61% HO: C 53.48, H 4.73, N 7.20, Cl 12.14).
Preparation 67 and Example 141.
(Scheme M: M-7 where R 3 is ethyl, R, is 4-[(2,6-dichlorobenzoyl)amino]phenyl, Y is and the stereochemistry is [2R-(2a,4p(S*)]-4-I[[1-Carboxy-2-[4-[(2,6-dichlorobenzoyl)aminolphenyllethyl]amino]-carbonyl]-1,2-azetidinedicarboxylic acid 1-ethyl ester (C 24
H
23 C1,NOs 8 To a suspension of M-6 (Example 140, Scheme M where R 3 is ethyl, R, is dichlorobenzoyl)amino]phenyl, Y is and the stereochemistry is (0.201 g, 0.347 mmol) in MeOH (5 mL) is added HO (4.3 mL)andl.00 M LiOH (0.70 mL). The reaction mixture is stirred at rt for 23 h. It is concentrated in vacuo. The aqueous concentrate is diluted with HzO, and the solution is adjusted to approximately pH 12 with IN NaOH. It is extracted with Et,O, and the EtO extract is discarded. The aqueous solution is adjusted to approximately pH 3 with IN HC1. It is extracted repeatedly with EtzO. The combined Et z O extracts were dried, filtered and concentrated to give, as a white solid, the title compound: TLC (500:500:1 MeOH/CH 2 C,1/HCO,H) Rf= 0.20; 'H NMR (CDOD) 8 7.59 7.50-7.37 7.23 4.78-4.64 4.61-4.51 4.22- 3.89 3.30-3.185 3.07-2.93 2.42-2.23 1.21 and 1.11 (3H total); MS (FAB) m/z 552.0946 (calcd for 552.0940); Anal. C 50.48, H 4.61. N 6.64, Cl 12.95 (calcd for 1.60% HO: C 51.35, H 4.31, N 7.49, Cl 12.63).
WO 99/67230 WO 9967230PCTIUS99/1 4233 -178- Scheme N.
X /)LOH .N-i
R
3 -Y-X ~~lOH N-2
R
H
2 N 0" N-3 0
R
0
R
0 L PN OH N-
R
3 0jH~
R
0
O
k) rl OHL N-6 Scheme N teaches a general method for the preparation of N-acvl azetidinecarboxylic acid Examples N-5 where n equals 0, 1 or 2, m equals 0, 1 or 2. (m n) equals 2, X is nitrogen and R 3 R, and Y are defined as in Scheme B, and azetidine-carboxylic acid Examples N-6 where n equals 0, 1 or 2. mn equals 0, 1 or 2, (mn n) equals 2, X is nitrogen and R, is defined as in Scheme B. Thus acylation of aminoacid N-I gives N-acylacid N-2, that is WO 99/67230 PCT/US99/14233 -179coupled with reagent N-3 (as exemplified by the use of reagents A-3 and A-4 of Scheme A, and B-4 and B-5 of Scheme B) to provide Examples N-4. Ester hydrolysis of N-4 provides Examples N-5. N-deacylation of Examples N-5 provides Examples N-6.
Preparation 68.
(Scheme N: N-2 where n is 2. m is 0. X is N, Y is -CO 2
R
3 is (1,1-dimethyl)ethyl, and the stereochemistry is (S)-l,2-Azetidinedicarboxylic acid 1-(l,l-dimethylethyl) ester (CH,,NO 4 To a mixture of (S)-(-)-2-azetidinecarboxylic acid (110 mg, 1.1 mmol), BocO (290 mg, 1.30 mmol), and DMAP (0.017 g, 0.14 mmol) in 4:1 DMF/H,O (10 mL) is added Et 3
N
(0.30 mL, 2.2 mmol). The reaction mixture is stirred at rt for 68 h. and then is concentrated. The concentrate is diluted with EtOAc, and the EtOAc solution is washed with cold 10% KHSO 4 The combined organic extracts are dried, filtered and concentrated to give the title compound as a colorless oil: TLC (750:250:1 Hexanes/acetone/HCO 2
H)
Rf 0.26; 1 H NMR (CD 3 OD) 6 4.97 4.57 3.98 3.87 2.57 2.13 1.42 MS (-ESI) m/z 200.3.
Preparation 69 and Example 142.
(Scheme N: N-4 where n is 2, m is 0, X is N, Y is -CO 2
R
3 is (1,1-dimethyl)ethyl, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is 4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-l-azetidinecarboxylic acid 1-(1,1-dimethyl)ethyl ester
(C
26
H
9
C
2
N
3 0 6 To a mixture of acid N-2 (Scheme N. where n is 2, m is 0, X is N, Y is R 3 is (1,1dimethyl)ethyl, and the stereochemistry is (1.04 g; 5.17 mmol) and HOBt-H,O (0.71 g, 5.3 mmol) in CH2C1 2 (10 mL) at 0 OC is added a mixture of EDC-HC1 (1.00 g, 5.22 mmol) in CH 2 C12 (20 mL). The reaction mixture is stirred at 0 °C for 30 min, and then N-3 (Scheme N where R, is 4 2 6 -dichlorobenzoyl)amino)phenyl and stereochemistry is (2.10 g, 5.20 mmol) and N-methylmorpholine (0.60 mL, 5.46 mmol) are added. The reaction mixture is stirred at 0°C for 30 min and at rt for 3 h. The reaction mixture is partitioned between 10% KHSO, and CH,C1l. The aqueous phase is extracted twice more with CH,Cl,. The combined organic extracts are washed with saturated NaHCO, and brine, WO 99/67230 WO 9967230PCT/US991 14233 -180an .d then are dried. filtered and concentrated to a yellow oil (2.54 g) that is purified by silica flash chromatography to give the title compound as a white foam: mp 106-108 'C; TLC 1 Hexanes/EtOAc); Rf= 0.21; [a]21D -38 (c 1.0 1, MeOH); 3 CNMR (CD3OD) 6 172.37, 171.58, 163.64, 156.68, 136.98, 136.22. 133.11, 131.87, 130.90, 129.49, 127.91, 120.00, 80.42, 61.87, 53.31, 51.45, 36.40, 27.16. 20.08; MIS (EI) m/z 551, 549, 478, 476, 451, 449, 396, 394, 351, 349, 280, 278, 175, 173; Anal. C 56.33, H 5.48, N 7.23, Cl 12.43 (calcd for 0.52% H,O: C 56.44, H 5.34, N 7.59. Cl 12.82).
Preparation 70 and Example 143.
(Scheme N: N-5 where n is 2, mn is 0, X is N. Y is R 3 is (1,1 -dimethyl)ethyl, R, is 4 2 ,6-dichlorobenzovl)amino]phenyl and stereochemnistry is [l1-Carboxy-2-14-I(2,6-dichlorobenzoyl)aminoI phenylljethyl] aminojcarbonylJ -1-azetidinecarboxylic acid 1 -dimethvlethyl) ester (C 25
H
27 C l,N 3 0 6 A solution of N-4 (Scheme N, where n is 2, in is 0, X is N, Y is R 3 is (11 dimethyl)ethyl, R, is 4 2 ,6-dichlorobenzoyl)aminojjphenyl and stereochemnistry is (507 mg, 0.92 mmol) and LiOH (1.25 mmol) in 1: 1 MeOH/H 2 0 (10 mL) is stirred at r1 for 18 h. The reaction mixture is diluted with cold 10% KHS0 4 and extracted with CH,C1 2 The organic extracts are dried, filtered and concentrated to a white foam (498 mg), that is purified by silica flash chromatography to give the title compound: TLC (750:250:1 hexanes/acetone/HCOH) Rf~= 0. 12; [c] 25 D -27 (c 0.94, CHCl 3 3 C NMR (CD 3 OD) 8 210.05, 172.70, 172.27, 163.63, 156.75, 136.89, 136.22, 133.36, 131.88, 130.90, 129.56, 127.91, 119.95, 80.47, 61.93, 53.18, 36.47, 27.15, 20.09; MS (-ESI) in/z 533.8;- MS (El) m/z 435, 419, 417, 401, 399, 373, 371, 280, 278, 175, 173, 147, 145; Anal. C 55.23, H 5.25, N 7.42, Cl 12.87 (calcd for 1. 10% H 2 0: C 55.36, H 5.14, N 7.75, Cl 13.07).
Preparation 71 and Example 144.
(Scheme N: N-6 where n is 2, mn is 0, X is N. R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemnistry is L) N-tI(2S)-2-Azetidinvljcarbonyll-4-[(2, 6-dichlorobenzoyl)aminoI-L-phenylalanine trifluoroacetic acid salt (C.,OH 19 C1 1
N
3 0 6 A solution of N-5 (Scheme N, where n is 2, mnis 0. X is N, Y is R 3 is 011 dimethyl)ethyl. R, is 4 2 6 -dichlorobenzovl)amino]phenyl and stereochemnistry is [2S- WO 99/67230 WO 9967230PCT[US99/I 4233 -181- (900 mg; 1.7 mmol) in 1: 1 TFA/ CH,C1 2 (5 mL) is stirred at rt for 1.5 h, and is concentrated. The residue is thrice diluted with CHCI 3 and re-concentrated. This residue is dissolved in MeOR and concentrated to a white foam, that is dissolved in 1: 1 MeOHJHO and then concentrated to remove most of the MeOH. The solution is frozen and lyophilized to give the product as a white powder: -6 (c 0.72. MeOH); 3 C NMR
(CD
3 OD) 8 172.62, 167.54, 163.78, 136.82, 136.13, 133.43, 131.84, 130.97, 129.38, 127.93, 120.15, 58.38, 54.03, 43.73, 36.27, 23.33; MS (+ESI) m/z 436.0; MS (FAB) m/lz 438, 436; Anal. C 46.77, H 3.75, N 7.24, Cl 12.44 (calcd for a 1: 1 TFA salt with 1.68% C 47.2 1, H 3.79, N 7.5 1, Cl 12.67).
Example 145.
(Scheme N: N-4 where n is 1. m is 1, X is N, R 3 is 1-dimethyl)ethyl, R, is 4- dichlorobenzoyl)amino]phenvl, Y is C0 2 and the stereochemistry is Ill-I 4-1R2,6-Dichlorobenzoyl)aminol phenyl] methyl]-2-methoxy-2oxoethyl] aminoj-carbonyll-l -azetidinecarboxylic acid 1 ,1-dimethyl)ethyl ester
(C
26
H
29 C1 2
N
3 0 6 Example 145 was prepared as described in Scheme N from 3-azetidinecarboxylic acid.
Physical properties as follows: TLC 1 EtOAc/hexanes) Rj 0.22; [a]I 2 'D +18 (c 0.92, MeOH); 1 3 CNMR (CD 3 OD) 8 172.92, 171.91, 163.68, 156.55, 136.87, 136.20, 133.27, 131.89, 130.91, 129.36, 127.90. 120.04, 79.80, 53.85, 51.45. 36.43, 32.26, 27.24; MS (El) m/z 469, 467, 451, 359, 351, 349. 280, 278, 175, 173. 57; Anal. C 56.82. H 5.39, N 7.52, C1 12.81 (calcd for 0.06% H,O: C 56.70, H 5.3 1, N 7.63, Cl 12.87).
Example 146.
(Scheme N: N-5 where n is 1, mn is 1, X is N, R 3 is 1-dimethyl)ethyl, R, 5 is 4-[2,6dichlorobenzoyl)amino~phenyl. Y is C0 2 and the stereochemnistry is 1 I-Ca rboxy- 2 -1 4 [(2,6-dichloro benzoyl)a min o Iphenyll ethyI amino] carbonyll I- 1 -azetidinecarboxylic acid 1 -dimethylethyl) ester (C,,H 27 C1 2
N
3 0 6 Example 146 was prepared from example 145 by the procedure described in preparation Physical properties as follows: TLC (600:400:1 Hexanes/Acetone/HCO,H) Rf 0. 17; [a] 2 0.92 MeH) 1 3 C NR( OD) 8 173.05, 172.89, 163 .69, 156.56, 136.78, 136.19, 133.59, 131.89, 130.91.,129.40. 127.90. 120.00. 79.78. 53.67, 51.72. 36.47, 32.3 1, WO 99/67230 WO 9967230PCT/US99/I 4233 -182- 27.24. MIS (-ESI) m/lz 533.9; MIS (FAB) m/z 538. 536, 438, 436, 337, 335. 280. 278, 175, 173. 57: Anal. C 55.03, H 5.21, N 7.52., CI 12.81 (calcd for 1.22% H 2 0: C 55.30, H 5.15, N 7.74. Cl 13.06).
Example 147.
(Scheme N: N-6 where nis 1, mis IlX is NH, R 5 is 4-[2,6-dichlorobenzoyl)amino]phenyL, and the stereochemnistry is 13-Azetidinyli carbonyll-4-I(2,6-dichlorobenzoyl)amino I-L-phenylalanine, trifluoroacetic acid salt (C 20
H,
9 C1 2
N
3 0 6
.C
2
HF
3 Example 147 was prepared from Example 146 by the procedure described in preparation 71. Physical properties as follows: 2 1D +32 (c 0.87. MeOH); 3 C NMR (CD.,OD) 6 173.32. 170.57, 163.80, 136.70, 136.13. 133.76, 131.84, 130.97, 129.45. 127.93, 120.17, 54.03. -36.59, 35.32); MS (FAB) m/z 438, 436.,391, 331, 175, 173, 101, 55: Anal. C 46.85. H 4.07, N 7.33, Cl 12.39 (calcd for a 1: 1 TFA salt with 3.34% H'0: C 46.4 1, H 3.92, N 7.38, Cl 12.45).
Example 148.
4 -[(2,6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2.
oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3--(2-pyridinylmethyl) ester (Scheme A, A-7: where RA.I and RA-2 are the same and equal to proton, R 3 is 2pyridinylmethyl, Y is R 5 is 4-[(2,6-dichlorobenzoyl)amino]phenyI and stereochemistry is
NN
N 0 c Example 148 was prepared as described in Scheme A from D-cysteine using 2pyridinemethanol to form the requisite carbamnate. Physical data as follows: 'H NMR (300 MHz. CDC13) 5 8.37 (1 7.70 (1 7.51 (2 7.27 (6 7.08 (2 6.92 (1 H), 5.24 (2 4.77 (3 4.40 (1 3.74 (3 3.37(1 3.15 (3 3 C NMR (75 MHz, WO 99/67230 PCT/US99/14233 -183- CDC1 3 8 171.4, 162.5, 155.2, 149.0, 137.1, 136.5, 136.0. 132.4. 130.7, 129.9, 128.0, 123.1, 120.6, 63.0. 53.4. 52.5. 36.8; MS (ESI+) for C 2 8
H,
6
CN
4 0, 6 S m/z 616.8 HRMS (EI) calcd for C,,H, 6 C1N 4 0 6 S 616.0950, found 616.0946. Anal. Calcd for
C
28
H
26 C12N 4 0 6 S: C. 54.46; H, 4.24; N, 9.07. Found: C, 54.61; H, 4.32; N, 8.97.
Example 149.
[S-(R*,R*)]-4-[[[I-Carboxy-2-[4-[(2,6-dichlorobenzoyl) amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(2pyridinylmethyl) ester (Scheme A, A-8: where and RA 2 are the same and equal to proton, R3 is 2-pyridinylmethyl, Y is R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (S,
H
s- o N N JOH N 0 0 cl o Example 149 was prepared from example 148 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 1713, 1666, 1605, 1576, 1561, 1539, 1515, 1442, 1431, 1413, 1351, 1325, 1271, 1194, 766cm-'; 'HNMR(300 MHz,CD 3 OD) 5 8.80 (1 H), 8.56 (1 7.99 (2 7.58 (2 7.45 (3 7.25 (2 5.43 (2 4.60 (3 3.30 (3 2.93 (2 13 C NMR (75 MHz, CD 3 OD) 8 172.7. 171.5, 163.7. 152.8, 151.3, 146.6, 141.9, 136.8, 136.1. 133.6. 131.8, 130.9, 129.6, 127.9, 126.1. 125.4, 120.1, 63.2, 62.8, 59.4, 53.5, 36.4, 35.1; MS (ESI+) for C 2 7 H24C12,N 4 0 6 S m/z 602.9 MS (FAB) m/z (rel. intensity) 603 59), 605 603 154 139 137 136 (47), 123 105 103 93 HRMS (FAB) calcd for C 2 7
,H,
4
C,N
4 0 6 S +H, 603.0872, found 603.0876; Anal. Calcd for C 2
,H,
4
C,N
4 0 6 S 0.3 H,O: C, 53.26; H, 4.07; N, 9.20. Found: C. 52.97: H, 4.23: N, 9.04.
WO 99/67230 PCTIUS99/14233 -184- Example 150.
2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]- 6 oxo-3-thiazolidinepentanoic acid (Scheme A, A-7: where RA., and RA-2 are the same and equal to proton. R, is (CH 2 3
CO,H,
Y is CO-, R, is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is s 0 H O O 0 o
HI
Example 150 was prepared as described in Scheme A from D-cysteine using glutaric anhydride to form the requisite amide. Physical data as follows: IR (mull) 3077, 3053, 3040, 1738, 1728, 1696, 1682, 1641, 1557, 1437, 1430, 1414, 1307, 1232. 1209 cm'; 'H NMR (300 MHz, CDC 3 8 7.56 (2 7.30 (3 7.08 (2 4.96 (1 4.62 (3 3.72 (3 3.28 (4 2.37 (4 1.90 (2 3 C NMR (75 MHz, CDCI 3 6 179.5, 176.0, 175.6, 173.6, 167.2, 140.6, 140.0. 136.1, 134.5, 133.7, 133.4, 131.8, 124.4, 66.7, 65.9, 57.2, 56.3, 40.3, 39.3, 37.3, 36.7, 36.2, 33.5. 23.6; MS (FAB) m/z (rel. intensity) 596 90), 598 597 596 341 263 230 225 193 (31), 141 88 HRMS (FAB) caled for C 2 6 H,,C2N30,S 596.1025, found 596.1036.
Anal. Calcd for C, 6 H,,Cl 2
N
3 0,S 0.3 H,O: C, 51.88; H, 4.62; N, 6.98. Found: C, 51.69; H, 4.69; N, 6.59.
Example 151.
[S-R*,R*)]-4-[[[1-Carboxy-2-[4-[(2,6-dichlorobenzoyl)amino] phenyl]ethyl]amino]carbonyl]-&-oxo-3-thiazolidinepentanoic acid methyl ester (Scheme A, A-8: where RA,1 and RA-2 are the same and equal to proton. R, is
(CH,)
3 CO,CH3, Y is CO-, R 5 is 4 2 6 -dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -185s o O QN OH o o 0 o 1 Example 151 was prepared as described in Scheme A from D-cysteine using methyl glutaryl chloride to form the requisite amide. Physical data as follows: IR (mull) 3287, 3196, 1724. 1662. 1607, 1562, 1540, 1516. 1431. 1414, 1326, 1268, 1217, 1195, 799 cm-'; MS (FAB) m/z (rel. intensity) 596 (MH, 72), 598 596 229 193 167 133 129 121 103 89 'H NMR (300 MHz, CD30D) 6 7.56 (2 7.43 (3 7.23 (2 4.53 (3 3.63 (3 2.96 (4 2.45 (3 2.24 (2 H), 1.93 (2 "C NMR (75 MHz, CDCI 3 5 174.0, 172.1, 169.9. 162.9, 162.8, 136.1, 132.2, 130.6, 130.0, 127.9, 120.3, 62.9, 54.5, 51.6, 49.8, 36.8, 33.5, 33.0, 32.7, 29.6, 19.6; HRMS (FAB) calcd for C, 6
H,
7 C1lN 3 O0S 596.1025, found 596.1047. MS (FAB) m/z (rel.
intensity) 596 72), 598 596 229 193 167 133 129 121 103 89 (99).
Example 152.
1 -Carboxy- 2 4 -[(2,6-dichlorophenyl)methoxy]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[2-(l-piperidinyl)ethyl] ester (Scheme A, A-8: where RA^. and RA.
2 are the same and equal to proton, R 3 is 2-(1piperidinyl)ethyl, Y is R 5 is 4 2 ,6-dichlorophenyl)methoxy]phenyl and stereochemistry is 0
SNOH
o° c, Example 152 was prepared from example 5 by the procedure described in preparation 6.
Physical data as follows: IR (mull) 3254. 2654, 1711. 1565. 1547, 1512, 1438, 1344.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -186- 1300. 1240, 1196, 1179, 1119. 1014, 767 cm-' 'H NMR (300 MHz. CD 3 0D) 8 7.45 (2 H), 7.36 (1 7.19 (2 6.97 (2 5.26 (2 4.50 (5 3.60 (11 1.83 (6 MS (ESI+) for C 38
H
33 C1 2
N
3 0 6 S m/z 610.0 Anal. Calcd for 8
H
33 C1,N 0 6 S 1.5 H 2 0' HCI: C, 49.90; H, 5.53; N. 6.24; Cl, 15.78. Found: C. 49.86, H. 5.43; N, 6.29; Cl, 15.65.
Water 3.99.
Example 153.
I -Carboxy-2-[4-[(2,6-dichlorophenyl)methoxrlphenyl] ethyl] amino]carbonyll-N-methyl-N-[2-(2-pyridinyl)ethyl]-3-thiazolidinecarboxamide (Scheme A, A-8: where and RA.
2 are the same and equal to proton. is 2-(2-pyridyl)ethyl, Y is CON(CH,)-, R, is 4-[(2,6-dichlorophenyl)methoxvjphenvl and stereochemistry is SA) (N (N flOH c Example 153 was prepared as described in Scheme A using 2-(2-methylaminoethyl)pyridine to formn the requisite urea and hydrolysis according to the procedure described in preparation 6. Physical data as follows: mp 80"C (softens), 125'C-. IR (mull) 1661, 1611, 1585, 1565, 1511, 1489, 1439, 1394, 1300, 1240. 1196,1179, 1017. 779, 768 1H NMR (300 MHz, CD 3 OD) 6 8.44 (1 7.75 (1 7.35 (5 7.12 (2 6.93 (2 H), 5.22 (2 4.83 (1 4.65 (1 4.32 (2 3.77 (1 3.45 (1 3.20 (1 3.00 2.84 (3 3 C NMR (75 MHz. CD 3 OD) 6 173.0, 170.8. 162.4. 158.4, 157.9, 147.9, 138.0, 136.6, 132.2. 130.6. 130.1, 129.3, 128.3, 124.2, 122.0. 114.5, 64.8, 64.7, 53.3, 52.5, 49.7, 35.7, 34.8, 32.8; MS (ESI+) for C, 9
H
30 IC1 2
N
4 0 5 5 m/z 617.0 Anal. Calcd for
C,
9
H
30 Cl2N 4 0 5 S: C, 56.40; H. 4.90; N, 9.07. Found: C, 56.3 1; H, 5.07; N, 8.98.
Preparation 72 and Example 154.
)I-4-III[1-[[ 4 2 .6-Dichlorobenzoyl)aminolphenvl]methyl]-2-[(4-pyridinyl)methoxy]-2-oxoethyllamino]carbony]-3-thiazolidinecarboxylic acid 3--ethyl ester WO 99/67230 PCT/US99/14233 -187- S- 0
SNH
N0 NH 0 N HN 0
C
NH
Cl6 To a solution of Example 12 (Scheme A. A-8: where and R, 2 are the same and equal to H, R 3 is ethyl, Y is CO 2
R
5 is 4-[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is (400 mg, 0.74 mmol) in dimethylformamide (4 mL) was added tetramethylguanidine (204 iL, 1.63 mmol) followed by 4-picolyl chloride (138 mg, 0.81 mmol). The solution was heated to 65 °C for 3 h and volatiles removed in vacuo.
Purification of the residue by flash chromatography using methylene chloride/methanol as eluant afforded the title compound (320 mg) as an amorphous solid: IR (mull) 3275, 1748, 1677, 1608, 1561, 1539, 1515, 1431, 1415, 1344, 1325, 1271, 1222, 1194, 799 'H NMR (300 MHz, CDC1 3 6 8.87 (1 8.48 (2 7.50 (2 7.25 (3 H), 7.10 (2 6.93 (2 5.08 (2 4.79 (2 4.61 (1 4.28 (1 4.13 (2 3.16 (4 1.21 (3 'C NMR (75 MHz, CDCI,) 6 174.6, 170.7, 170.6, 162.6, 149.2, 144.5, 136.9, 136.0. 132.3, 131.7, 130.7, 129.8, 128.0. 122.5, 120.3, 65.0, 63.0, 62.7, 53.3, 37.3, 20.9, 14.5; MS (ESI+) for C 2 9 H,,Cl 2
N
4 06S m/z 630.8 Anal. Calcd for
C
29
H
2 8 Cl 2
N
4 0 6 S: C, 55.15; H, 4.47; N, 8.87. Found: C, 54.85; H, 4.58; N, 8.74.
Anal. Calcd for C, 9
H
28
C
2
N
4 0 6 S: C, 55.15; H. 4.47; N, 8.87; Cl, 11.23; S, 5.08.
Found: C, 54.85; H, 4.58; N, 8.74.
Preparation 73 and Example 155.
-Carboxy- 2 -[4-[(2,6-dichlorophenyl)methoxy]phenyl]ethyl]amino]carbonyl]-8-methyl- -thia-4,8-diazaspiro[4.5]decane-4-carboxylic acid 4-ethyl ester (Scheme B, B-7: where and RB., are the same and equal to H, RB.
3 and R.4 together form a cyclic ring of 6 atoms of the formula -CHCH,N(CH 3 )CHCH,-, Y is CO,, R 3 is ethyl, R, is 4-[(2,6-dichlorophenyl)methoxy]phenyl and stereochemistry is WO 99/67230 WO 9967230PCTIUS99/1 4233 -188-
H
3 C- V.J"N N -IOH N.I 0 N t-Butyl ester B-6 (Scheme B where RB., and RB.
2 are the same and equal to H, 3 and RB.
together form a cyclic ring of 6 atoms of the formula -CH,CH,N(CH,)CH 2
CH
2 Y is
CO
2
R
3 is ethyl, RB.5 is O-t-butyl, R, is 4-[(2,6-dichlorophenyl)methoxylphenyl and stereochemnistry is prepared according to Scheme B from 1 -methyl-4-piperidone and D-cysteine) (681 mg, 1.02 mmol) was dissolved in a solution of HC1 in dioxane (4 M, 28 mL) at ambient temperature. After 18 h, volatiles were removed in vacuo to afford a residue (650 mg) which was lyophilized from water. Further purification of a portion of this product (200 mg) was effected by chromatography on a Biotage Flash 40' system using a 40 g KP-C 1 8-HS (3 5-70 gim) silica gel cartridge using aqueous acetonitrile as eluant to afford the title compound (94 mg) as an amorphous powder: IR (mull) 1696, 1611, 1585, 1565, 1511, 1439, 1404, 1335, 1303, 1271, 1239, 1196, 1178, 1017, 769 cm I NMR (DMSO-d 6 3 00 MHz) 8 8.25 (1 7.54 (2 7.43 (1 7.13 (2 6.94 (2 5.16 (2 4.84 (1 4.41 (1 4.00 (2 3.07 (4 2.73 (4 2.53 (3 1.96 (1 1.69 (1 1. 11 (3 13 C NMR (CDCl 3 75 MHz) 6 173.9.,169.2, 157.7, 153.2, 136.9, 132.1, 130.7, 130.5. 129.3, 128.5, 114.7, 73.2, 66.7, 65.2, 62.8, 54.0, 43.5, 36.5, 14.6; MS (ESI+) for C, 8
H
33 C1,-N 3 0 6 S mlz 610.0 MIS for C 28
H
33 0-1 2
N
3 0 6
S
rnlz 607.9 HRMS (FAB) calcd for C 28
H
33
C,N
3 0 6 S+H, 610.1545, found 610.1561.
Anal. Calcd for C 28
H
33 C1,-N 3 0 6 S 0.6 HCI H 2 0: C, 51.70; H, 5.52; N, 6.46; Cl, 14.17.
Found: C, 51.28; H, 5.49; N, 6.50; Cl, 14.57. Water 2.72.
Example 156.
4 2 6 -Dichlorophenyl)methoxy]pheny]methyl]2....methoxy.2oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-(3-tetrahydrofuranyl) ester (Scheme A, A-7: where and RA.
2 are the same and equal to proton. R 3 is 3tetrahydrofiiranyl, Y is R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl and stereochemistry is WO 99/67230 PCT/US99/14233 -189-
O
o° C l Example 156 was prepared as described in Scheme A from D-cysteine using 3hydroxytetrahydrofuran to form the requisite carbamate. Physical properties as follows: mp 125-126.5 0 C. IR (mull) 3311, 1750, 1744, 1708, 1661, 1549, 1515, 1439, 1408, 1307, 1243. 1227, 1212, 1173, 1019 cm'; 'H NMR (300 MHz, CDOD) 8 7.38 (3 7.14 (2 H), 7.96 (2 5.20 (1 5.25 (2 4.61 (4 3.79 (4 3.74 (3 3.19 (2 2.84 (2 2.29 (2 'C NMR 75 MHz, CDC13, spectra complicated via the presence of diastereomers) 5171.7, 171.6, 158.1, 137.0, 132.1, 130.5, 130.4, 130.3, 128.5, 128.2, 115.1, 77.2, 77.2, 73.2, 67.0, 65.2, 63.1, 63.0, 53.3, 52.5, 52.4, 36.9, 32.9; MS (ESI+) for
C
26 H,C12N 2 0 7 S m/z 582.8 Anal. Calcd for C 26
H
2 8
C
2
N
2 0 7 S: C, 53.52; H, 4.84; N, 4.80. Found: C, 53.34; H, 4.87; N, 4.86.
Example 157.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-hexahydro- y-oxo- 1 H-azepine- 1butanoic acid
NH
N OH HO
CI
HO,
0 0 0 C1 Example 157 was prepared as described for the preparation of Example 167. Physical properties as follows: IR (mull) 1781. 1709. 1651, 1625, 1612, 1550, 1537. 1515, 1444, 1431. 1418. 1398, 1331, 1193, 798 'H NMR (300 MHz, DMSO-d 6 5 1.45 (8 H), 2.30 (4 2.90 (3 3.80 (1 4.50 (2 7.16 (2 7.50 (5 7.94 (1 10.62 WO 99/67230 WO 9967230PCTIUS99/1 4233 -190- (1 7.71 (1 MIS (FAB) m/lz (rel. intensity) 578 43), 581 580 579 5 78 577 227 226 198 173 98 (46).
Preparation 74 and Example 158.
(Scheme N, N-6: where n is 2, m is 0, X is N, R, is 4+(2.6dichlorobenzoyl)amino]phenyl and stereochemnistry is 2S-(R 12S-(R*,R*)I-2-[[Ill-I 4 -(2,6-Dichlorobenzoyl)aminolphenyll methyII-2-methoxy-2oxoethyll amino] carbonylljazetidine
(C
2
,H
2
CI,N
3 0 4 A solution of the product of example 142 (Scheme N: N-4 where n is 2, m is 0, X is N, Y is R 3 is (1,1-dimethyl)ethyl, R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl and stereochemnistry is (512 mg; 0.93 mmol) in 1: 1 TFAICHC., (10 mL) is stirred at r1 for I h. The reaction mixture is concentrated under reduced pressure. The residue is taken up in a mixture of CH,C1, and satd aqueous NaHCO 3 The aqueous phase I extracted twice additionally with CH,Cl,. The combined CH,C1, portions are dried, filtered and concentrated to give a yellow oil (440 mg), that is purified by silica flash chromatography (95:5 CH 2 Cl 2 /MeOH) to afford the title compound (324 mg) as a white foam: mp 113- 115 OC; TLC (95:5 CH2ClIMeOH) Rf= 0. 10; [cz] 2 5 D -34 (c 0.96, MeGH); UV (MeOH) X, 224 (sh, F. 12100), 251 (17700), 284 (sh, 2880); IR (mineral oil mull) 3260, 1744, 1664, 1606. 1561, 1537, 1515, 1431, 1414, 1323. 1270. 1223, 1195, 799, 782 'H NMR (CD 3 OD) 8 7.61 7.50-7.3 8 7.25 4.86 (1 4.75 (1IH), 4.21 (1 H), 3.73 3.61 3.39-3.28 (1IH), 3.23 (1 3.05 (1IH), 2.63-2.50 (1IH), 2.21-2.08 (I1H); MIS (+ESI) m/z 450.0; MIS (El) m/z 451, 449, 396, 3 94, 351, 349, 278, 211, 175, 173, 96, 70, 56; Anal. C 55.68, H 4.79, N 8.96, Cl 15.43 (calcd for 1.08% H 2 0: C 55.4 1, H 4.77, N 9.23, Cl 15.58).
Example 159.
O-[(2,6-Dichlorophenyl)methy]-N-[[(4S)-3 -methylsulfonyly....
thiazolidinyl]carbonyl]-L-tyrosinamide (Scheme C, C-10: where Rc- 2 Rc- 3 and k4 are the same and equal to proton, R 3 is methyl, Y is R, is 4 2 6 -dichlorophenyl)methoxv]phenyl and stereochemnistry is WO 99/67230 WO 9967230PCTIUS99/1 4233 -191- H H O=S=OO0C CI6 Example 159 was prepared as described in Scheme C using methanesulfonyl chloride to form the requisite sulfonamide. Physical properties as follows: mp 228-230IC; 'H NMR (300 MHz, DMSO-d 6 8 8.03 (1 7.54 (2 7.44 (2 7.14 (3 6.92 (2 5.16 (2 4.69 (2 4.41 (1 4.31 (1 3.21 (1 3.01 (3 2.87 (3 3 C NMR MHz, DMSO-d 6 8172.9, 169.1. 157.5. 136.5, 132.3, 132.0, 130.8, 1'30.6. 129.2, 114.6, 65.3, 64.4, 54.3, 52.0, 37.3, 35.0; MIS (ESI-) for C 21
H
23
C,N
3 0 5 m/z 530.2 Anal.
Calcd for C,,H 23 C1,N 3 0 5 5 2 C, 47.37; H. 4.35; N, 7.89. Found: C, 47.43; H, 4.46; N, 7.81.
Example 160.
4 2 ,6-Dichloropheny)methoxy]phenyl]methyl]-2methoxy.2.
oxoethyl]amino]carbonyl] -l -thia-4-azaspiro [4.4]nonane--4-carboxylic acid 4-ethyl ester (Scheme B, B-6: where and RB.
2 are the same and equal to H, RII- and 4 together form a carbocyclic ring of 5 atoms, is OCH 3 Y is CO 2
R
3 is ethyl, R 5 is dichlorophenyl)methoxylphenyl and stereochemnistry is 0 a ci~ Example 160 was prepared as described in Scheme B. Physical properties as follows: IR (mull) 1746, 1705, 1681, 1510, 1439, 1399, 1336, 1301, 1276. 1241, 1203. 1178, 1110, 17, 769 'H NMR (CDCI 3 8 7.36 (2 7.25 (1 7.03 (2 6.93 (2 6.64 (1 5.22 (2 4.85 (2 4.15 (2 3.73 (3 3.15 (4 2.69 (1 2.48 (1 H), 1.76 (6 1.23(3 3 CNMR (CDC1 3 5 171.5, 170.7, 158.0, 152.5, 137.0, 132.0, 130.5.,130.3, 128.5. 128.2, 115.0, 66.4. 65.2, 61.9, 53.1, 52.4, 37.1. 32.3, 25.1. 24.6, 14.5; MS (ESI+) for C 2
,H
32 C1,N,0 6 S m/z 594.9 MS (ESI-) for C, 28 H.12CI,N.,O 6 S M/Z WO 99/67230 WO 9967230PCTIUS99/1 4233 -192- 592.8 Anal. Calcd for C.,,H 32 C,N,0 6 S 0. 10 H,O: C, 56.30; H, 5.43:- N, 4.69.
Found: C. 56.20: H. 5.24; N, 4.69. Water 0.3 1.
Example 161.
I -Carboxy-2-[4-[(2,6dichlorophenyl)methoxy]phenyllethyl]amino]carbonyl]..i-thia-4-azaspiro 4 .4]nonane-4carboxylic acid 4-ethyl ester (Scheme B. B-7: where and R- 2 are the same and equal to H, RB-3and RE,4together form a carbocyclic ring of 5 atoms, YS is 2
R
3 is ethyl, R, is 4 -12,6-dichlorophenyl)methoxylphenyl and stereochemnistry is 0 C, 0 ci~ Example 161 was prepared from example 160 by the procedure described in preparation 6.
Physical properties as follows: IR (mull) 1737, 1708, 1675, 1612, 1511, 1439, 1402, 1338, 1301, 1241,.1197, 1179,1115, 1018, 769 NMR (DMSO-d 6 )8 8.07 (1 H), 7.54 (2 7.45 (1 7.12 (2 6.94 (2 5.15 (2 4.63 (1 4.33 (1 3.91 (2 3.05 (2 2.79 (1 2.60 (1 1.60 (6 1.07(3 3 C NMR (DMSO-1 6 6 172.7, 169.4. 157.1, 155.7, 135.9, 131.6; 13 1.4. 130.3, 129.6, 128.7, 114.1, 83.9, 64.7, 60.5. 53.1. 38.0, 36.2, 31.8, 24.2, 24.1, 14.1; MS (ESI+) for C 2 7
H
30 Cl1 2
N
2 0 6 5 m/z 580.8 MS (ESI-) for C 2 1 7
H
30 C1N 2 0 6 5m/z 578.8 HRMS (El) calcd for
C
27
H
30 C1.,N,10 6 S 580.1202. found 580.1172; Anal. Calcd for C 27
H
30 C,N,0 6 S 0. 19 H 2 0: C, 55.44; H, 5.24; N, 4.79. Found: C, 55.24; H. 5.32; N, 4.79. Water 0.59.
Preparation 0
H
2
NI.Q.'CH
3
HCI
HI The aminoester product of preparation 75 is useful as a synthetic intermediate (for example, reagent A-4 of Scheme A).
WO 99/67230 PCT/US99/14233 -193- To a cold (0-5 0 C) solution of anhydrous methanolic HCI was added 100 g of L-4nitrophenylalanine (Advanced ChemTech) portionwise over 15 min. The mechanically stirred mixture was heated to gentle reflux for 48 h. The mixture was allowed to cool and then filtered through a sintered glass filter funnel, washing the collected solids with hot MeOH until only insoluble residues remained. The filtrate was concentrated in vacuo to afford the methyl ester (120 g) as waxy off white solid which was used without further purification.
To a suspension of methyl ester described above (87 g, 0.33 mole) in CH 2 Cl 2 (1500 mL) at ambient temperature was added di-t-butyldicarbonate (109 g, 0.50 mole) followed by the dropwise addition of Et 3 N (51 mL. 0.37 mole). After 15 min additional Et 3 N (40 mL, 0.29 mol) was added to maintain a slightly basic mixture (ca. pH The reaction mixture was stirred 18 h and additional CHC1, (1400 mL) and Et 3 N (15 mL, 0.11 mol) were added.
After an additional 2 h the reaction mixture was quenched by the slow addition of MeOH (100 mL), stirred for 1 h and then partitioned between CH 2 C12 and cold 10% aqueous
KHSO
4 The organic layer was washed with saturated NaHCO 3 and brine, dried (Na 2
SO
4 filtered and concentrated in vacuo. Flash chromatography of the residue using hexane and a gradient of a 1:1 mixture of EtOAc/CHCl, (25-33%) afforded the Boc-methyl ester (69 g) as a white solid. Physical properties as follows: 'H NMR (300 MHz; CDC1 3 8 8.16 7.31 5.04 4.63 3.73 3.18 1.41 MS for CiH 20
N
2 0 6 m/z 325.2 Palladium on carbon (10% w/w, 1.25 g) was added to a Parr hydrogenation flask under a N, atmosphere and carefully wetted with 100 mL of MeOH/THF A solution of the Boc-methyl ester described above (25 g, 77 mmol) in 400 mL of MeOH/THF was added and the mixture shaken on a hydrogenation apparatus under a hydrogen atmosphere (20 psi) for 1 h at ambient temperature. The reaction mixture was filtered through a pad of Celite and the solids washed several times with MeOH. The combined filtrates were concentrated in vacuo to afford the 4-aminophenylalanyl derivative (22.7 g) which was used without further purification. Physical properties as follows: 'H NMR (300 MHz, CDC13) 8 6.89 6.61 4.96 4.50 3.69 2.95 1.41 MS for C,,HN,,0 m/z 295.2 WO 99/67230 PCTIUS99/1 4233 -194- A cold solution of 2,6-dichlorobenzoyl chloride (11.1 mL, 77.5 mmol) in 125 mL of THF was treated dropwise with a solution of the 4-aminophenylalanyl derivative described above (22.7 g, 77.1 mmol) and Et 3 N (16 mL, 115 mmol) in 125 mL of THF.
The reaction mixture was allowed to warm to temperature and stir an additional 18 h. The mixture was diluted with EtOAc (2 L) and then washed with 1N HC1, H,O, IN NaOH and brine. The organic extract was dried (NaSO,), filtered, and concentrated in vacuo to give the crude product as a pale yellow solid. This material was recrystallized from acetone/hexanes (ca. 1:1) to afford the amide (30.8 g) as a crystalline solid. Physical properties as follows: mp 192.2-193.1°C; IR (mull) 3305, 1747, 1736, 1690. 1665, 1609, 1548. 1512. 1433, 1414, 1325, 1277, 1219. 1171. 781 'H NMR (300 MHz; CDCl 3 8 7.57 7.34 7.14 4.98 4.60 3.74 3.11 1.42 (9H); MS for C22H 2 4
CI
2
N
2 0 s m/z 467.0 To 650 mL of anhydrous 4M HCI in dioxane at ambient temperature was added the amide described above (30.6 g, 65.5 mmol) portionwise and the resulting mixture was stirred until all the solids had dissolved (ca. 1 Volatiles were removed in vacuo to give a light yellow solid which was partitioned between water (500 mL) and ether (1 The water layer was separated and concentrated in vacuo to approximately 200 mL. The aqueous solution was then frozen and lyophilized to afford the aminoester product (25.6 g) as a light yellow solid. Physical properties as follows: [oa] 25 D +5 (c 1, MeOH); IR (mull) 3244,3186.3112, 1747, 1660, 1604, 1562, 1539. 1516, 1431, 1416, 1327, 1273, 1243, 799 cm'; 'H NMR (300 MHz; CD 3 OD) 5 7.69 7.45 7.29 4.34 3.83 3.21 "C NMR (300 MHz; CD3OD) 6 169.0, 163.9, 137.8, 136.08, 131.8, 131.0, 130.3, 129.7, 127.9, 120.6, 53.8, 52.3, 35.4; MS for C,,H, 6 CI2N,0 3 m/z 367.1 Preparation 76.
H
2 N O.CH 3
HCI
Cl 1 WO 99/67230 PCT/US99/14233 -195- The aminoester product of preparation 75 is useful as a synthetic intermediate (for example, reagent A-4 of Scheme A).
To a cold (0-5 0 C) solution of anhydrous methanolic HCI (200 mL) was added 25 g of Nac-t-Boc-O-2,6-dichlorobenzyl-L-tyrosine (Sigma) portionwise over 15 min. After minutes at 0-5 0 C, the mixture was heated to 50 0 C for 2 h. The solution was cooled to room temperature and the volatiles removed in vacuo. The solid was suspended in ethyl ether and collected by filtration to afford the title compound (21.4 g) which was used without further purification. Physical properties as follows: [ca] 2 5 D +16(c 1.00, ethanol); 'H NMR (300 MHz. CD30D) 6 7.44 (2 7.35 (1 7.21 (2 7.02 (2 H), 5.28 (2 4.29 (1 3.81 (3 3.18 (2 MS (ESI+) for C,H-IC12NO 3 m/z 354.1 Anal. Calcd for C, 7
H,
7
CI
2 NO- HCI: C, 52.26; H, 4.64; N, 3.59. Found: C, 52.17; H, 4.74; N, 3.61.
Example 162 S)-1-[[4-[(2,6-Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-l-piperidinecarboxylic acid 1-[(1,1-dimethyl)ethyl] ester H O
O
HO
O
H0 BocI 0 oa Ex.162 Example 162: HCI gas was bubbled through a solution of N-(tert-butoxycarbonyl)-4-(2,6dichlorobenzoylamino)-L-phenylalanine (2.51g, 5.53 mmol) in MeOH (20 mL) for minutes. The solution was stirred for additional 2 hours at room temperature. The solvent was removed in vacuo and the excess HCI was removed by the addition of Et20 (3 x mL) and evaporation under reduced pressure. The resultant gum was dissolved in THF mL) and N-tert-butoxycarbonyl-pipecolinic acid (1.28 gm, 5.59 mmol), BOP-reagent WO 99/67230 PCT/US99/14233 -196- (2.69 gm, 6.09 mmol) and DIEA (2.9 mL. 16.6 mmol) were added and the reaction mixture was stirred overnight. EtOAc (25 mL) was added and the mixture was extracted with IN HCI (20 mL). The organic phase was washed with saturated LiCI (20 mL) then saturated NaHCO 3 (30 mL). The organic layer was dried over Na 2
SO
4 filtered and evaporated. Chromatography of the residue (silica gel, Hexanes 50% EtOAc/Hexanes gradient elution) provided Example 162 as a solid (1.45 gm, 45 ESMS 578, 580 (MH Example 163 4 2 6 -Dichlorobenzoyl)amino]phenyl]methyl]-2-methoxy-2oxoethyl]amino]carbonyl]-y-oxo- 1 -piperidinebutanoic acid Ha- NH 0 NH O- H I NH Lu I NExample 163 Cl HCI gas was bubbled through a MeOH (20 mL) solution of Example 162 (1.27 gm, 2.20 mmol) for 10 min. Stirring was continued overnight at room temperature. The solvent was removed in vacuo and the excess HCI was removed by washing with Et20 (3 x 10 mL) on a vacuum filter. The HCI salt was completely dried under high vacuum to provide Compound 1 (1.09 gm, 97 as a solid: ESMS 478, 480 Example 163: Compound 1 (147 mg, 0.285 mmol) was dissolved in DMF (5 mL) containing DIEA (150 pL, 0.88 mmol). To this solution was added succinic anhydride (59 mg, 0.59 mmol) and the mixture was stirred at 50 0 C for 5 hr under dry nitrogen. The solvent was evaporated and the residue was purified by column chromatography (silica gel, Hexanes EtOAc gradient elution) to provide Example 163 as a solid (164 mg): ESMS 578, 580 WO 99/67230 WO 9967230PCT/US99/I 4233 -197- Examples 164-166 The following mono methyl esters were prepared in a similar manner as Example 163 0
NH..AI
N
P 0-C N H
R
3 HOO9-e HOOC 0
HOOCLJ
MS
(m/Z) 646
(MH+)
620
(MH+)
574 Example 167 2- 1 I -Carboxv-2- [4-[(2,6-dichlorobenzoyl )aminolphenyl] ethyl] amino]carbonyl] -yoxo- 1 -piperidinebutanoic acid
H-O
0 0 0 CI N H b Example 167 Example 163 (154 mg, 0.266 mmol) was treated with LiOH (26 mg, 1.07 mmol) in
H
2 0 (5 mL) for 3 hours. The product was then precipitated by the addition of 3 N HCL.
WO 99/67230 WO 9967230PCTIUS99/I 4233 -198- The product was collected by vacuum filtration and washed with cold H 2 0 (2 x 3 mL).
Drying under high vacuum provided Example 167 as a solid (109 mg): ESMS (mlz) 562 Example 168-170 The following compounds were prepared in a similar manner as 167.
OH
0 C1
NH
C1 HOOKo
HOOCL."
MS
(m/z) 630 604 560 Preparation 77 c N Mlerrifield resin Intermediate -1 0 =po~mtr WO 99/67230 WO 9967230PCTIUS99/1 4233 -199- Intermediate-1: Attachment of N-tert-butoxycarbonv I- [4-(2,6,-dichlorobenzovi amino+].
L-phenylalanine to Merrifield resin was done using Horiki's method (Horiki et al., Chem.
Lett. 1978(2) 165-168). In a 250 mL round bottom flask fitted with a drying tube, Merrifield resin (Biorad. 10.0 g, 13.5 mmol/g) and anhydrous potassium fluoride (Aldrich, 1 .57g, 27.0 mmol) were added to a solution of N-tert-butoxycarbonyl-[4-(2,6,dichlorobenzoylamino)]-L-phenylalanine (Bachem California, 6.13 g, 13.5 mmol) in dry DMF (100 mL). The reaction mixture was stirred at 80*C in an oil bath for 24 hr. The cooled resin was then filtered and washed thoroughly wvith DMF (2 x 250 mL), aqueous DMF (3 x 250 mL), methanol (3 x 250 mL). dichloromethane (3 x 250 mL), and finally methanol (3 x 250 mL). The resin was then dried under reduced pressure to constant weight to give Intermediate-I. Incorporation ofN-tert-butoxycarbonvl-[4-(2,6,dichlorobenzoylamino)]-L-phenylalanine onto the resin was estimated to be 0.045 mmol/g from the increase in resin mass.
Example 171 1,3-Benzodioxol-5-yl)-l1-methyl-5-oxo-3-pyrrolidinylcarbonyl]-4-[(2,6dichlorobenzoyl)amino]-L-phenylalanine 0 0 N O RB N0 0 H 0r 0N N0 1 N OH 0 0 C1 Intermediate- I C1 Exami 171 'N N
H
C I Example 171: The Intermiediate-i (0.15 g, 0.1065 mmnol/g) was pretreated with CH 2
CI
2 (2x 3 mL). The swollen resin was then deprotected with 50% TFAICH 2 Cl 2 G(3mL, 30mmn). The resin was rinsed in the following order: CH 2
CI
2 (2 x 3 mL), CH 3 0H (2 x 3 mL). CHCl-, (2 x 3 mL). The resin was swollen with DMF (2 x 3 mL). 2-(3.4- WO 99/67230 PCT/US99/14233 -200methylenedioxyphenyl)-1-methyl-5-oxo-3-pyrrolidine carboxylic acid (84 mg, 0.32 mmol) in DMF (1.0 mL) was activated with 0.5 M HBTU/HOBT in DMF (0.7 mL) and DIEA (0.139 mL, 0.799 mmol), and then added to the swollen resin. The mixture was vortexed for 2 hr at room temperature. The resin was filtered and washed in the following order: DMF(2 x 3 mL), CH 2 Cl 2 (2 x 3 mL), CH 3 OH (2 x 3 mL), CH 2 Cl 2 (2 x 3 mL), respectively. If a Kaiser test on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resulting resin was then dried in vacuo to constant weight. The resin was placed in the polypropylene column and pretreated with 3 mL of THF. Then to the preswollen resin 1.6 mL of THF, 0.48 mL of CH3OH, and 0.160 mL of 2N LiOH were added. The mixture was vortexed for 15 min and filtered to a clean and preweighed test tube. The resin was next washed with 2mL of
CH
3 0H (2x) and the combined filtrates were evaporated. The resulting gum was dissolved in 1 mL of water. The solution was then acidified with IN HC1 to pH 2.0. The precipitate was centrifuged, washed with water (5mL, 2x) and dried in vacuo to furnish 15.4 mg of Example 171 as a solid. ESMS 596, Preparation 78 O.Y NH BOC. xi NH 0 Cn Intemedite- I Intenediate-2 NH C, C, b~ WO 99/67230 PCT/US99/14233 -201- D E 0 F G CN)-'tr- N -ND 0 0 N
X
I
H
J K O O O Intermediate-2F: The resin bound N-tert-butoxycarbonyl-[4-(2,6dichlorobenzoylamino)]-L-phenylalanine (Intermediate-i), (250 mg, 0.1125 mmol/g) was placed in a 8.0 mL, polypropylene filter column fitted with a 2-way polypropylene stopcock. The resin was pretreated with CH 2 Cl 2 (2 x 3 mL). The swollen resin was then deprotected with 50% TFA/CH 2
CI
2 (3-4 mL, 30min) with shaking. The resin was rinsed in the following order: CH 2
CI
2 (2 x 3 mL), CH 3 0H (2 x 3 mL), CH 2 C1 2 (2 x 3 mL).
The resin was swollen with DMF (2 x 3 mL). N-Tert-butoxycarbonyl-nipecotic acid (103 mg, 0.45 mmol) in DMF (1.0 mL) was activated with 0.5 M HBTU/HOBT in DMF (0.910 mL) and DIEA (0.195 mL), then added to the swollen resin. The mixture was vortexed for 2 hr at room temperature. The resin was washed in the following order: DMF (2 x 3 mL),
CH
2 Cl 2 (2 x 3 mL), CH 3 0H (2 x 3 mL), CH 2 C1 2 (2 x 3 mL) and dried (Intermediate- 2F). If a Kaiser test on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained.
The intermediate resins Intermediate-2D, 2E, 2G, 2H, 21, 2J 2K were each produced following this procedure.
Example 172 S)-1-Carboxy-2-[4-[(2,6dichlorobenzoyl)amino]phenyl]ethyl]amno]carbonyl] -1-piperidinyl]carbonyl]-3pyridinecarboxylic acid WO 99/67230 PCT/US99/14233 -202- H O
OH
O O NN NR N O H N 0 ocl COOH
C
Intermediate-2D Example 172 Example 172: Resin bound Intermediate-2D (2.0 g, 1.3 mmol/g) was pretreated with
CH
2
CI
2 (2 x 20 mL). The swollen resin was then deprotected with 50% TFA/CH 2
CI
2 mL, 30min). The resin was rinsed in the following order: CH 2 C12 (2 x 20 mL), CH 3
OH
(2 x 20 mL), CH 2
CI
2 (2 x 20 mL). The resin was swollen with DMF (2 x 20 mL). Pyridine dicarboxylic acid (652 mg, 3.9 mmol) in 20 mL of DMF was activated with M HBTU/HOBT in DMF (8.0 mL) and DIEA (1.7 mL, 9.75 mmol), then added to the swollen resin. The mixture was vortexed for 2 hr at room temperature. The resin was filtered and washed in the following order: DMF(2 x 20 mL), CH 2 Cl 2 (2 x 20 mL),
CH
3 0H (2 x 20 mL), CH 2 C12 (2 x 20 mL), respectively. If a Kaiser test (Kaiser et al., Anal. Biochem. 1970, 34, 594-598) on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resulting resin was then dried in vacuo to constant weight (2.2 The resin was treated with 25 mL of liquid HF by stirring for 60 min at 0°C in an HF-reaction apparatus (Peninsula Laboratories Inc., Belmont, CA). The HF was rapidly evaporated off by vacuum aspiration at 0°C. Then 100 mL of dry ethyl ether was added. The resin and the resulting precipitates were filtered off and washed three times with 50 mL of ethyl ether, and dried in vacuo. The mixture was then treated with 25 mL of IN NaOH and the combined solutions were lyophilized. The crude product was purified by HPLC using a C-18 column and a linear acetonitrile/0. 1% HCI gradient. The gradient was run from 60% solvent A HCI) to WO 99/67230 WO 9967230PCT/US99/1 4233 -203solvent B (80% acetonitrile in 0. 1% HCl) in 20 min. Lyophilization furnished 20 mg of Example 172. ESMS (mlz): 612 Examples 173-267 0 0 Bocx.NH.., ,JRB Q, NH, NH
NH
Method-A (Example 173) 4-[(2,6-Dichlorobenzoyl)amino]-N-[ [1-(3-methoxy-l1-oxopropyl)-3 -piperidinylicarbonyl]- L-phenylalanine 0 0 C1 Example 173 The Intermediate-2F (0.25 g, 0. 1125 mmol/g) was pretreated with CH 2 Cl 2 (2 x 3 mL).
The swollen resin was then deprotected with 50% TFAfCH 2 (3mL. 30mmn). The resin was rinsed in the following order: CH 2
CI
2 (2 x 3 mL), CH 3 0H (2 x 3 mL), CH1 2 Cl 2 (2 x 3 mL). The resin was swollen with DMF (2 x 3 mL). 3-methoxypropionic acid (53 mg, 0.45 mxnol) in DMF (1.0 mL) was activated with 0.5 M HBTU/HOBT in DMF (0.9 mL) and DIEA 195 mL), then added to the swollen resin The mixture was vortexed for 2 hr at room temperature. The resin was filtered and washed in the following order: DMF WO 99/67230 PCT/US99/14233 -204- (2 x 3 mL), CH 2 Cl 2 (2 x 3 mL), CH 3 0H (2 x 3 mL), CH 2
CI
2 (2 x 3 mL), respectively. If a Kaiser test on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resulting resin was then dried in vacuo to constant weight. The resin was placed in the polypropylene column and pretreated with THF (3 mL). Then THF (3.5 mL), CH 3 0H (1.0 mL) and 2N LiOH (0.175 mL) were added. The mixture was vortexed for 15 min and filtered to a clean and preweighed test tube. The resin was next washed with THF/5% CH 3 0H (2mL) and the combined filtrates were evaporated. The resulting gum was dissolved in H 2 0 (1 mL). The solution was then acidified with IN HCI to pH 2.0. The precipitate was centrifuged, washed with water (2 x 5 mL) and dried in vacuo to furnish 38.3 mg of Example 173 as a solid: ESMS 548 Method B (Example 174): -Carboxy- 2 4 -[(2,6-dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-yoxo-1-piperidinebutanoic acid HON r N -'OH 0 0 Cl Example 174 The Intermediate-2F (0.25g, 0.1125 mmol/g) was pretreated with CH 2 C12 (2 x 3 mL). The swollen resin was then deprotected with 50% TFA/CH 2
CI
2 (3mL. 30min). The resin was rinsed in the following order: CH 2 Cl 2 (2 x 3 mL), CH 3 0H (2 x 3 mL), CH 2 Cl 2 (2 x 3 mL). The resin was then swollen with DMF (3mL). Succinic anhydride (45 mg, 0.45 mmol) dissolved in DMF (4 mL) was added to the swollen resin and stirred at for 2hr. The resin was filtered and washed in the following order: DMF (2 x 3 mL),
CH
2 Cl 2 (2 x 3 mL), CH 3 0H (2 x 3 mL). CH 2
CI
2 (2 x 3 mL), respectively. If a Kaiser WO 99/67230 PCT/US99/14233 -205test on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resulting resin was then dried in vacuo to constant weight. The resin was placed in the polypropylene column and pretreated with THF (3 mL). Then to the swollen resin THF (3.5 mL), CH 3 0H (1.0 mL) and 2N LiOH (0.175 mL) were added. The mixture was vortexed for 15 min and filtered to a clean and preweighed test tube. The resin was next washed with THF/5% CH 3 OH (2 x 2 mL) and the combined filtrates were evaporated. The resulting gum was dissolved in H 2 0 (1 mL).
The solution was then acidified with IN HCI to pH 2.0. The precipitate was centrifuged, washed with H 2 0 (2 x 5 mL) and dried in vacuo to furnish 30.5 mg of Example 174 as a solid: ESMS 562 Method C (Example 175): 4 -Carboxyphenyl)amino]carbonyl]-4-piperidinyl]carbonyl]-4-[(2,6dichlorobenzoyl)amino]-L-phenylalanine H rN OH 0O CI 0
HIS,
Example 175 The Intermediate-2G (0.25g, 0.1125 mmol/g) was pretreated with CH 2 Cl 2 (2 x 3 mL). The swollen resin was then deprotected with 50% TFA/CH 2
CI
2 (3mL, 30min). The resin was rinsed in the following order: CH 2
CI
2 (2 x 3 mL), CH 3 OH (2 x 3 mL), CH 2 Cl 2 (2 x 3 mL). The resin was then swollen with DMF (3mL). Ethyl 4-isocyanatobenzoate (22 mg, 0.108 mmol) dissolved in DMF (3 mL) and DIEA (47 iL 0.27 mmol) were added to the swollen resin. This reaction mixture was vortexed for 6-8 hr at room temperature.
The resin was filtered and washed in the following order: DMF (2 x 3 mL), CH 2 Cl 2 (2 x 3 mL), CH 3 0H (2 x 3 mL), CH 2 C1 2 (2 x 3 mL), respectively. If a Kaiser test on a small WO 99/67230 PCT/US99/14233 -206quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resulting resin was then dried in vacuo to constant. The resin was placed in the polypropylene column and pretreated with THF (3 mL). Then to the swollen resin THF (3.5 mL), CH 3 OH (1.0 mL) and 2N LiOH (0.175 mL) were added respectively.
The mixture was vortexed for 15 min and filtered to a clean and preweighed test tube.
The resin was next washed with THF/5% CH 3 OH (2mL) and the combined filtrates were evaporated. The resulting gum was dissolved in H 2 0 (1 mL). The solution was then acidified with IN HCI to pH 2.0. The precipitate was centrifuged, washed with H 2 0 (2 x mL) and dried in vacuo to furnish 18 mg of Example 175 as a solid: ESMS 625, Examples 176-266 The following compounds were prepared in a similar manner as described above.
OH
N 0 CI ex# Method Q X, MS (m/z) 176 A HOOC ,560 0
Q
N
178 A COOH 616 S1' o" 179 A 560
N
o WO 99/67230 WO 9967230PCTIUS99/1 4233 -207- 1-80 A vc 0-626 Iscojo 1 181 A 0 610 182 A 0 610 0-J
COON
183 A 0'A660 Hoocll 0([M-Hf) 184 A HOac. 0 611 N-I Io 185 A 0C567 C0Y,- 186 A 0 611 N0
HOOC
187 A 0 611 HOOC .1 188 A OH 633 C N 0 189 A HOOC H600 'I N 0 190 A HOOC 621 HN0 191 A 02N 603 0 192 A 560 1o 193 A 560 0 14 BHOOC-Al 562 195 B 0630 HOOC D(M-] 196 B 0604 HOOC N WO 99/67230 WO 9967230PCT/US99/1 4233 -208- 197 B 0Oj~ 56 198 A HOO40.~ 560 0 199 A fbco-N \O 548 200 A COOH 07~ 616 0 N 201 A 00560 0 202 C HOOC,N.. 563 0 203 A fb 00 626 204 A 0 0 610 akco, 205 A 0610 QA. 206 A 0 0660 NOc, 207 A 0611 208 C HOOYhoC-~ 625 H BHOOC-'k N& 562 210 B 0630 HOOC fO 211 B 604 HOOC 0C 212 B HOC 0 560 213 A 0N~- 560 214 A Fbcol-ro 0N-- 548 WO 99/67230 WO 9967230PCTIUS99/1 4233 -209- 215 A COCH 0Nj~4 616 216 A 00560 0 217 C H OOCI..JJN 563 0 218 A 0 coNQ0J( 626 bco ND 219 A 610 C OH 220 A 0 610 Ql"-
COOH
221 A Ooj N o660
([M-HP)
222 A 0 0N~- 1 23 CH0OC -NC 625 N 224 B HOO'. 0X H 224 B 61654 226 B 0~1 590 227~~ B Ho~~ 4 00C 228 A 546 229 A 0 546 230 A OO 0 I 231 A HOC.:~ 546 o 232 C OO H 0 549 0 WO 99/67230 WO 9967230PCTIUS99/1 4233 -210- 233 A i. 0N 0612 ISCO (MH- 234 A 0 0 596 cOH (EM-Hp) 235 A 0 0596
COON
236 A 0X, 646 1
HOOCJ
237 A 0 ('to0 597 HOOC~f/~([M-H]) 238 C HOOO, 0 N 0 611 H I' 239 A 0 Ii 571 N 240 A 0 -615 I
HOOC
241 A 0 s-615 HOOCIN 242 A O 3 1 0 243 A HOOC Hs604 11 N'N N-'-If 0 244 A HOW C1 s 625 0 245 A s-564 O 246 A 0 -564 0 247 A 0562 HOOCQKN> 24 A 00 WO 99/67230 PCTIUS99/1 4233 -211- 250 A 611 N 0r 251 A 611 0 0 252 A 0 567 N 0 253 A UY< 560 0 0 254 A 548 OOC
M-H)
255 A 546 255 A 0 256 A 534 H3CO 257 A Hr 0o597 NU 258 A 1" 59 HOOA N 0 259 A 0 553 N KT~ 260 A 546 261 A 0 564 H k([M-Hp 262 A 0 552 263 A 0o, <6 HOO,,,NZ 615 264 A rT 615 HOOC N ;[MTH0 0 1 265 A 00S- 571 266 A 564 0 WO 99/67230 WO 9967230PCTIUS99/1 4233 -212- Example 267 S)-1 -Carboxv.
2 4 -[(2,6-dichlorobenzoyl)amino~phenvl]ethyljamnino]carbonyI]..
tetrahydro-y-oxo-1I,4-thiazepine-4(SH-1)-butanoic acid H 0 -H 0 0 N RB YN .RB 0 CI 0 0 C I H ,+H Intermediate- I CI Iiitermediate-3
CI
S-
NY N ORB N 0 HO o~ 0 ci o N N I~ Example 267 Nl The Intermediate-I (0.3g, 0. l95mmollg) was pretreated with CH 2 Cl 2 (2x 3 mL). The swollen resin was then deprotected with 50% TFAICH,CI, (3mL, 30 min). The resin was rinsed in the following order: CH 2 C1, (2x 3 mL), CHOH (2x 3mL), CH,Cl-,(2x 3mL). The resin was swollen with DMF (2x 3mL). carboxylic acid (204 mg, 0.78 mmol) in DMF (2.0 mL) was activated with 0.5 M HBTU/HOBT in DMF (1.6 mL) and DIEA (0.340 niL. 1.95 mmnol), then added to the swollen resin. The mixture was vortexed for 2 hr at room temperature. The resin was filtered and washed in the following order: DMF (2x 3 mL), CH,C1 2 (2x 3mL), CH- 3 0H (2x 3mL), CH 2 Cl 2 (2x 3mL), respectively. If a Kaiser test on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resin (intermediate-'), 0.2 g, 0. 13 mmol/g) was pretreated with CHC 2 (2x 3 mL). The swollen resin was then deprotected with 50% TFAICI-I,CI. (3mL. 30 min). The resin was rinsed in the following order: CH-,CI, (2x 3mL), CHOH (2x 3mL), CHCI 2 (2x 3mL). The resin was then swollen with DMF (3mL). Succinic anhydride (78 mig, 0.78 mmol) WO 99/67230 PCT/US99/14233 -213dissolved in DMF (4mL) was added to the swollen resin and stirred at 50C for 2hr. Then the resin was filtered and washed in the following order: DMF (2x 3mL), CH,C1 2 (2x 3mL), CH 3 OH (2x 3mL), CH,Cl, (2x 3mL) respectively. If a Kaiser test on a small quantity of the resin is positive (blue) then repeat the coupling procedure until a negative result is obtained. The resulting resin was then dried in vacuo to constant weight. The resin was placed in the polypropylene column and pretreated with THF (3 mL). Then THF (3.9 mL), CH3OH (1.2 mL) and 2N LiOH (0.195 mL) were added to the swollen resin. The mixture was vortexed for 15 min and filtered to a clean and preweighed test tube. The resin was next washed with THF/5% CH 3 OH (2x 2mL) and the combined filtrates were evaporated. The resulting gum was dissolved in H,O (1 mL). The solution was then acidified with IN HCI to pH 2.0. The precipitate was centrifuged, washed with H,O (2 x and dried in vacuo to furnish 80 mg of Example 267 as a solid: ESMS (m/z) WO 99/67230 WO 9967230PCT/US99/1 4233 -2 14- Scheme 0
QOH
Wang resin I 0 Fmoc-HN OH 0-2 000NH-FmocN0 N 0-3 0 2
N!-
0 0 0 NH-Fmoc 0-4
H
2
N!
0 Q0 NH-Fmoc
N
H
0 0 0 NH2 0-6
H
WO 99/67230 WO 9967230PCTLUS99/1 4233 -2 Scheme 0 (continued) 0
NH
2 0-6 H
R
0
RO
0 3 0 R -1RA- N1 0-10 0-11
R
0 3 0 O-4k SR 0 HO H N 0 2 -R 3
H
WO 99/67230 PCT/US99/14233 -216- Where: Ro., is defined as R 1 2
R-
2 is defined as C,.
6 alkyl or C 7 1 7 arylalkyl; Ro3, Ro4, and Ro.5, are defined independently as Ro.
6 is defined as R 2 Y, and Y 2 are defined independently as Y.
Scheme O describes a method for the preparation of examples of the formula 0-9 and 0-12. Commercially available Wang resin is acylated with commercially available N-a-Fmoc-Phe(NO 2 )-OH under standard conditions to afford the resin of formula 0-3. Reduction of the aromatic nitro group (Meyer et al., Mol. Diversity 1995, 1, 13-20) affords the resin bound aniline which may be reacted with a variety of electrophilic reagents to afford resin bound amides (0-5 where Y, is ureas where Y, is sulfonamides (0-5 where Y, is SO 2 and carbamates (0-5 where Y, is Removal of the Fmoc group under standard conditions provides amine of general structure 0-6 which is acylated using standard solid-phase peptide synthesis conditions (Atherton, Sheppard R.C. Solid Phase Peptide Synthesis: A Practical Approach; IRL Press at Oxford University Press: Oxford, 1989) with a commercially available or readily prepared thiazolidine-4-carboxylic acid of general formula 0-7 to afford the resin bound intermediate 0-8. Mild cleavage under standard conditions (Atherton, Sheppard R.C. Solid Phase Peptide Synthesis: A Practical Approach; IRL Press at Oxford University Press: Oxford, 1989) affords the acid of general structure 0-9.
In those cases where Ro2 is a 9 -fluorenylmethyl group, standard Fmoc group removal affords the amine of general structure 0-10, which may be reacted with a variety of electrophilic reagents as described in Scheme A to afford resin bound amides, ureas, sulfonamides and carbamates of general structure 0-11. Mild cleavage under standard conditions affords the acid of general structure 0-12.
Preparation 79 and Example 268.
(4S)-4-[[[(1S)-l-Carboxy-2-[4-[(benzoyl)amino]phenyl]ethyl]amino]carbonyl]-3thiazolidinecarboxylic acid 3-ethyl ester WO 99/67230 PCT/US99/14233 -217-
OH
sC 0
H
To a mixture of Wang resin DVB, Advanced Chemtech, 2.75 g, 2.20 mmol based on manufacture's loading of 0.8 mmol/g resin) in DMF (12 mL) was added N-ca- Fmoc-Phe(N0 2 0-2 (Advanced Chemtech, 1.90 g, 4.40 mmol) at room temperature.
After mixing for 10 min (by passing a slow stream of nitrogen through the mixture) pyridine (587 iL, 7.26 mmol) and 2,6-dichlorobenzoyl chloride (630 uL, 4.40 mmol) were added. The mixture was agitated overnight via nitrogen bubbling, filtered, washed with DMF, methylene chloride and methanol and dried in vacuo. In order to cap any unreacted hydroxymethyl groups, the resin was suspended in dichloroethane (5 mL) and to this mixture was added benzoyl chloride (0.75 mL) and pyridine (0.75 mL). The mixture was agitated for 2 h, filtered, washed with DMF, methylene chloride, methanol, methylene chloride and methanol, and dried in vacuo to afford the resin 0-3 (3.30 IR (diamond anvil) 1733, 1606 (resin), 1520 (resin), 1494 (resin), 1452 (resin), 1347, 1247, 1174, 1029 cm' To the prewashed (2 X 20 mL DMF) resin 0-3 (1.0 g, ca. 0.6 mmol based on an adjusted loading of 0.6 mmol/g) was added SnC1,* 2 H2O (6 mL of a 2M solution in DMF, 12 mmol). The viscous suspension was agitated for 4 hours by nitrogen bubbling, filtered and washed with DMF (2 X 20 mL). The resin was resuspended with SnCI 2 2 H 2 0 (6 mL of a 2M solution in DMF, 12 mmol), agitated overnight via nitrogen bubbling, filtered, washed extensively with DMF, water, 2-propanol, methylene chloride and methanol and dried in vacuo to afford the resin 0-4. Examination of the FTIR spectra of a small sample of resin 0-4 failed to exhibit an absorption at 1347 cm To a mixture of prewashed (2 X 20 mL CH 2 Cl 2 resin 0-4 (0.30 g, ca. 0.18 mmol based on an adjusted loading of 0.6 mmol/g) in 1,2-dichloroethane (3 mL) was added benzoyl chloride (174 gL, 1.50 mmol) and DIEA (313 gL, 1.80 mmol). The mixture was agitated overnight via nitrogen bubbling, filtered, washed with methylene chloride, DMF, methanol, and methylene chloride and dried in vacuo to afford resin 0-5. To a mixture of WO 99/67230 PCT/US99/14233 -218resin 0-5 in methylene chloride (5 mL) was added a solution of piperidine in DMF mL). A slow stream of nitrogen was bubbled through the mixture to effect mixing for min. The resin was filtered, washed with DMF and resuspended in a solution ofpiperidine in DMF 10 ml). After gentle mixing for 40 min, the resin was filtered and washed with DMF, methylene chloride, methanol and methylene chloride and diluted with DMF mL). To this mixture was added N-ethoxycarbonyl-D-thiazolidine-4-carboxylic acid 0.15 g, 0.72 mmol), HOBt (0.11 g, 0.72 mmol), PyBOP (0.37 g, 0.72 mmol) and DIEA (313 4L, 1.80 mmol). The reaction was mixed for 4 h at which point the qualitative Kaiser test was negative. The resin was filtered and washed with DMF, methylene chloride and MeOH and dried in vacuo to afford resin 0-8. After swelling with a minimum of methylene chloride (ca. 0.5 mL), the resin 0-8 was suspended with 95 aqueous TFA (5 mL). The mixture was mixed by magnetic stirring for Ih, filtered and washed with TFA (2 X 3 mL) and methylene chloride. The combined filtrates were evaporated in vacuo to afford a residue that was purified by flash chromatography using methylene chloride/methanol containing glacial acetic acid as eluant to afford the title compound (80 mg). Lyophilization from glacial acetic acid afforded an amorphous powder: IR (drift) 3311, 3298, 1670, 1601, 1579, 1531, 1487, 1412, 1380, 1345, 1324, 1265, 1205, 1190, 709 'H NMR (300 MHz, DMSO-d6) 8 10.39 (1 H), 8.39 (1 8.14 (2 7.86 (2 7.74 (3 7.36 (2 4.83 (2 4.59 (1 4.48 (1 4.22 (2 3.48 (2 3.26 (1 3.06 (2 1.35 (3 "C NMR (75 MHz, DMSOd 6 8 173.4, 169.8, 165.8, 154.0, 138.0, 135.4, 133.3, 131.9, 129.8, 128.8, 128.0, 120.4, 61.8, 54.2, 48.8, 36.7, 14.8; MS (ESI+) for C 23
H
2 5
N
3 0 6 S m/z 472.0 MS (ESI-) for
C
23
H
25
N
3 0 6 S m/z 470.1 MS (FAB) m/z (rel. intensity) 472 99), 472 (99), 371 160 81 71 69 57 55 43 41 HRMS (FAB) calcd for C 23
H
25
N
3 0 6 S 472.1542, found 472.1563; Anal. Calcd for
C
2 3
H
2 5
N
3 0 6 S 1.5 H 2 0: C, 55.41; H, 5.66; N, 8.43. Found: C, 55.47; H, 5.21; N, 8.00.
Example 269.
-Carboxy-2-[4-[(acetyl)amino]phenyl]ethyl]amino]carbonyl]-3thiazolidinecarboxylic acid 3-ethyl ester WO 99/67230 PCT/US99/14233 -219- <sT H o so 0 N N OH N CH3
H
The title compound was prepared as described in Scheme 0 using acetyl chloride to form the requisite amide. Physical data as follows: IR (drift) 3311, 1709, 1667, 1602, 1536, 1517, 1412, 1378, 1344, 1321, 1266, 1218, 1185, 1116, 769 cm'; 'HNMR (300 MHz, DMSO-d 6 5 9.79 (1 8.15 (1 7.37 (2 7.02 (2 4.54 (2 4.31 (1 H), 4.19 (1 3.95 (2 3.12 (1 2.93 (1 2.75 (2 1.82 (3 1.09 (3 "C NMR MHz, DMSO-d 6 8 173.2, 169.8, 168.5, 154.0, 138.2, 132.4, 129.8, 119.1, 62.1, 61.7, 53.9, 36.7, 24.4, 21.5, 14.8; MS (ESI+) for C, 8
H
23
N
3 0 6 S m/z 410.0 MS (ESI-) for
C,,H
23
N
3 06S m/z 408.0 MS (FAB) m/z (rel. intensity) 410 (MH 99), 486 411 410 409 205 188 177 160 148 88 HRMS (FAB) calcd for C, 8
H
23
N
3 0 6 S 410.1385, found 410.1379. Anal. Calcd for C,1H 23
N
3 0 6 S 0.3 H20: C, 52.11; H, 5.73; N, 10.13. Found: C, 51.73; H, 5.73; N, 9.82.
Example 270.
-Carboxy-2-[4-[(3-phenylpropanoyl)amino]phenyl]ethyl]amino]carbonyl]- 3-thiazolidinecarboxylic acid 3-ethyl ester
H
SOH
The title compound was prepared as described in Scheme 0 using hydrocinnamoyl chloride to form the requisite amide. Physical data as follows: IR (drift) 3311, 2978, 2930, 1665, 1601, 1534, 1517, 1413, 1379, 1344, 1252, 1216, 1187, 1115, 700 'H NMR (300 MHz, CDC1 3
/CD
3 OD 8 7.34 (2 7.21 (5 7.02 (2 4.67 (3 H), 4.30 (1 4.09 (2 3.11 (4 2.97 (2 2.58 (2 1.19 (3 "C NMR (75 MHz, CDCl 3 8 172.7, 171.3, 170.0, 154.8, 140.7, 137.0, 131.6, 129.7, 128.4, 128.2, 126.2, 119.9, 63.0, 62.6, 53.1, 38.9, 36.7, 31.5, 29.6, 14.2; MS (ESI+) for C 25
H
29
N
3 0 6 S m/z 500.2 WO 99/67230 PCT/US99/14233 -220- MS (ESI-) for C 25
H
29
N
3 0 6 S m/z 498.3 Anal. Calcd for C 25
H
29
N
3 0 6 S: C, 60.10; H, 5.85; N, 8.41. Found: C, 59.85; H, 6.07; N, 8.09.
Example 271.
(4S)-4-[[[(1S)-l-Carboxy-2-[4-[(3-pyridinylcarbonyl)amino]phenyl] ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester s o H O ^o 0 0 H N The title compound was prepared as described in Scheme O using nicotinoyl chloride to form the requisite amide. Physical data as follows: IR (drift) 3301, 3061, 2983, 2935, 1709, 1675, 1603, 1535, 1517, 1415, 1380, 1345, 1326, 1204, 1140 'H NMR (300 MHz, DMSO-d 6 8 12.74 (1 10.42 (1 9.11 (1 8.76 (1 8.29 (2 7.67 (2 7.55 (1 7.19 (2 4.62 (2 4.44 (1 4.29 (1 4.05 (2 3.24 (1 3.04 (1 2.88 (2 1.15 (3 3 C NMR (75 MHz, DMSO-d6) 173.1, 170.0, 164.3, 154.0, 152.4, 149.0, 137.7, 135.9, 133.5, 131.1, 129.8, 123.9, 120.5, 62.4, 61.8, 53.8, 36.7, 14.8; MS (ESI+) for C 22
H,
4
N
4 0 6 S m/z 473.3 MS (ESI-) for
C
22
H
24
N
4 0 6 S m/z 471.3 HRMS (FAB) calcd for C22H 24
N
4 0,S 473.1494, found 473.1509.
Example 272.
(4S)-4-[[[(1S)-1-Carboxy-2-[4-[(4-methoxybenzoyl)amino]phenyl]ethyl]amino]carbonyl]- 3-thiazolidinecarboxylic acid 3-ethyl ester
S--
OH
OHOCH
The title compound was prepared as described in Scheme O using p-anisoyl chloride to form the requisite amide. Physical data as follows: IR (drift) 1709, 1667, WO 99/67230 PCT/US99/14233 -221- 1604, 1532, 1514, 1439, 1412, 1379, 1343, 1323, 1255, 1221, 1178, 1027, 763 cm-; 'H NMR (300 MHz, CDC 3
/CD
3 OD 8 7.79 (2 7.47 (2 7.05 (2 6.87 (2 H), 4.68 (2 4.58 (1 4.28 (1 4.07 (2 3.79 (3 3.66 (2 3.04 (4 1.17 (3 3 C NMR (75 MHz, CDC1 3 8 177.9, 174.0, 170.1, 166.3, 158.8, 141.1, 135.8, 133.7, 133.0, 130.8, 124.6, 117.7, 66.9, 66.6, 59.3, 40.7, 33.5, 24.5, 18.2; MS (ESI+) for
C
24
H
27
N
3 0 7 S m/z 502.0 MS (ESI-) for C 24
H
2
,N
3 0 7 S m/z 500.1 MS (FAB) m/z (rel. intensity) 502 52), 503 502 297 240 160 135 88 73 69 57 HRMS (FAB) calcd for C 24
H
27
N
3 0 7 S +H, 502.1648, found 502.1657. Anal. Calcd for C 24
H
2 7
N
3 0OS 0.3 H 2 0: C, 56.86; H, 5.49; N, 8.29. Found: C, 56.65; H, 5.34; N, 7.92.
Example 273.
(4S)-4-[[[(1S)-l-Carboxy- 2 4 -[(4-methylbenzoyl)amino]phenyl]ethyl]amino]carbonyl]-3thiazolidinecarboxylic acid 3-ethyl ester N N'-OH
CH
3 The title compound was prepared as described in Scheme O using p-tolouyl chloride to form the requisite amide. Physical data as follows: IR (drift) 3310, 2981, 2929, 1671, 1608, 1599, 1531, 1517, 1413, 1379, 1344, 1324, 1265, 1210, 1188 'H NMR (300 MHz, DMSO-d 6 5 10.10 (1 8.24 (1 7.85 (2 7.67 (2 7.33 (2 H), 7.17 (2 4.62 (2 4.41 (1 3.28 (2 4.02 (2 3.23 (1 3.04 (1 2.87 (2 2.38 (3 1.15 (3 3 C NMR (75 MHz, DMSO-d 6 6 173.2, 169.8, 165.6, 154.0, 141.9, 138.1, 133.1, 132.5, 129.7, 129.7, 128.1, 120.4, 62.4, 61.8, 54.0, 36.7, 21.4, 14.8; MS (ESI+) for C2 4
H
27
N
3 06S m/z 486.2 HRMS (FAB) calcd for C 24
H
27
N
3 06S +H, 486.1699, found 486.1713; Anal. Calcd for C 2 4
H
2 7
N
3 0 6 S 0.3 H 2 0: C, 58.71; H, 5.67; N, 8.56. Found: C, 58.37; H, 5.67; N, 8.35.
WO 99/67230 WO 9967230PCT/US99/1 4233 -222- Example 274.
(4 IS)-l1-Carboxy-2- 2 -(trifluoromethyl)benzoyl]aminojphenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester N N OH ~O~OO0 CF,
N"-
H
The title compound was prepared as described in Scheme 0 using (2trifluoromethyl)benzoyl chloride to form the requisite amide. Physical data as follows: IR (drift) 3295, 1709, 1663, 1603, 1533, 1518, 1414, 1380, 1344, 13 16, 1269, 1176, 1132, 1108, 769 cm'; 'H NMR (300 MHz, DMSO-d 6 5 10.48 (1 8.23 (1 7.77 (2 H), 7.58 (2 7.17 (2 4.64 (2 4.44 (1 4.29 (1 3.99 (2 3.24 (1 3.05 (2 1.09 (3 3 C NMR (75 MHz, DMSO-d 6 8 173.2, 169.2, 165.8, 154.1, 137.8, 136.7, 133.0, 130.4, 129.9, 128.9, 126.7, 126.4, 126.0, 119.8, 62.2, 61.8, 54.0, 37.1, 14.7; MS (ESI±) for C 2 4
H
2 5
F
3
N
3 0 6 5 m/z 540.0 MS (ESI-) for C 24
H
25
F
3
N
3 0 6 5 m/z 538.1 (M- HRMS (FAB) calcd for C 24
H
24
F
3
N
3 0 6 S 540.1416, found 540.1423; Anal. Calcd for C 24
H
24
F
3
N
3 0 6 S 0.5 H 2 0: C, 51.49; H, 4.70; N, 7.83. Found: C, 51.42; H, 4.42; N, 7.45.
Example 275.
[[[(15)-l1-Carboxy- 2 4 -[(2,4,6-trichlorobenzoyl)amino]phenyl~ethyl]amino..
carbonyl]-3-thiazolidinecarlboxylic acid 3-ethyl ester 0 0 c Ca i WO 99/67230 PCT/US99/14233 -223- The title compound was prepared as described in Scheme O using 2,4,6trichlorobenzoyl chloride to form the requisite amide. Physical data as follows: IR (drift) 3286, 2926, 1709, 1664, 1604, 1578, 1542, 1517, 1413, 1379, 1345, 1325, 1269, 1218, 1187 'H NMR (300 MHz, CDCl 3
/CD
3 OD 8 7.58 (2 7.41 (2 7.20 (2 4.84 (2 4.69 (1 4.41 (1 4.19 (2 3.27 (4 1.26 (3 3 C NMR MHz, DMSO-d 6 173.0, 169.8, 161.5, 154.0, 137.2, 135.9, 135.1, 133.9, 132.6, 130.1, 128.5, 119.6, 62.2, 61.7, 54.0, 36.7, 14.8; MS (FAB) m/z (rel. intensity) 574 (MH 576 574 160 91 88 69 57 55 43 41 (39); HRMS (FAB) calcd for C 23
H
22
CI
3
N
3 0 6 S 574.0373, found 574.0364. Anal. Calcd for
C
23
H
2 2
CI
3
N
3 0 6 S: C, 48.06; H, 3.86; N, 7.31; Cl, 18.50. Found: C, 48.52; H, 4.13; N, 7.08.
Example 276.
(4S)-4-[[[(1S)-1-Carboxy-2-[4-[[(2,5-dichlorophenyl)sulfonyl]amino] phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester
N-
0H C 0 Cl The title compound was prepared as described in Scheme O using dichlorobenzene sulfonyl chloride to form the requisite sulfonamide. Physical data as follows: IR (drift) 1709, 1676, 1531, 1512, 1450, 1428, 1409, 1378, 1344, 1221, 1167, 1143, 1113, 1101, 1041 cm-; 'H NMR (300 MHz, DMSO-d 6 5 8.36 (1 7.90 (3 H), 7.20 (2 7.12 (2 4.70 (2 4.47 (1 4.34 (1 4.10 (2 3.19 (4 1.27 (3 3 C NMR (75 MHz, DMSO-d6) 8 173.0, 169.8, 153.9, 138.7, 135.5, 134.8, 134.1, 132.6, 131.0, 130.5, 130.0, 120.0, 62.4, 61.7, 53.5, 36.3, 14.8; MS (FAB) m/z (rel.
intensity) 576 99), 652 578 577 576 160 106 88 81 69 57 HRMS (FAB) calcd for C 22 H2 3 C1 2
N
3 0OS2+H, 576.0433, WO 99/67230 PCT/US99/14233 -224found 576.0400. Anal. Calcd for C 22
H
2 3 Cl 2
N
3 0 7
S
2 0.1 H 2 0: C, 45.70; H, 4.04; N, 7.27.
Found: C, 45.94; H, 4.04; N, 6.87.
Example 277.
(4S)-4-[[[(1S)-1-Carboxy-2-[ 4 -[[[(2,6-dichlorophenyl)amino]carbonyl] amino]phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester
OH
0^ C1 H H
CI
The title compound was prepared as described in Scheme O using 2,6dichlorophenyl isocyanate to form the requisite urea. Physical data as follows: IR (drift) 3284, 3277, 1709, 1655, 1600, 1569, 1544, 1452,1431,1415,1347,1238,1217, 1195, 771 cm'; 'H NMR (300 MHz, DMSO-d 6 5 9.09 (1 8.41 (1 8.09 (1 7.52 (2 H), 7.31 (3 7.08 (2 4.62 (2 4.35 (1 4.28 (1 3.27 (1 3.02 (1 2.85 (2 1.10 (3 "C NMR (75 MHz, DMSO-d 6 5 173.6, 169.8, 154.0, 152.8, 138.7, 134.4, 133.9, 131.3, 130.0, 128.8, 128.7, 118.1, 62.4, 61.8, 54.3, 36.7, 14.8; HRMS (FAB) calcd for C 23
H,
4 C1N 4 0 6 S 555.0872, found 555.0877. Anal. Calcd for C 23
H
24 C1 2
N
4 0 6 S 2 HzO: C, 46.71; H, 4.77; N, 9.47. Found: C, 47.08; H, 4.53; N, 9.06.
WO 99/67230 WO 9967230PCT[US99/1 4233 -225- Scheme P
-OH
Wang resin Boc-HN OHP-2 0 0 NH-Boc P-3 0 Q 0 NH-Bac P-4
HI
Rp(N
N
0 0
H
2 N
OH
IH
0 0
H
2 N 0 r CH 3
H
N, N.P 0 WO 99/67230 WO 9967230PCTIUS99/I 4233 -226- Scheme P (continued) tP-3 S Rp dA. P-7
R
N A R H 0 0'p- 0 0Rp.3 0 H R;-4 S R
N
0 N R- 0
H
0 0 R~ Rp- 3 H R3 0 0 Rp -3 HO
N
H N 0 2 R 0 0 R p- H 5 Rpz HO N jN'R- H 0 0)0' R- 0 P-1 0 P-1 1 WO 99/67230 PCTIUS99/1 4233 -227- Where: Rp., is defined as R, 2 Rp.
2 is defined as C,, 6 alkyl or C 7 1 arylalkyl; Rp.3, Rp.
4, and are defined independently as R.
6 is defined as R 2 Scheme P describes a method for the preparation of examples of the formula P-9 and P-12. Commercially available Wang resin is acylated with commercially available N-ca-Boc-Phe(I)-OH under standard conditions to afford the resin of formula P-3. Carbonylation of the resin bound aryl iodide with carbon monoxide and an amine in the presence of a source of palladium(0) affords the resin bound amide of general formula P-5 (for a general review of carbonylation chemistry, see: ('alquhoun, H.M.; Thompson, Twigg, M.V. Carbonylation Plenum Press: New York, 1991). Mild cleavage under standard conditions affords the amino acid of general structure P-5 which is esterified under mild acid catalysis to afford the amino ester of general structure P-6.
Condensation with a commercially available or readily prepared thiazolidine-4-carboxylic acid of general formula P-7 under conditions described in Scheme A affords the pseudodipeptide of general structure P-8. Mild base hydrolysis of the ester of general structure P-8 affords the acid of general structure P-9. In the case where Rp-2 is a 9fluorenylmethyl group, standard Fmoc group removal (Atherton, Sheppard R.C. Solid Phase Peptide Synthesis: A Practical Approach; IRL Press at Oxford University Press: Oxford, 1989) affords the amine of general structure P-10, which may be reacted with a variety of electrophilic reagents as described in Scheme A to afford amides, ureas, sulfonamides and carbamates of general structure P-11. Mild base hydrolysis of the ester in general structure P-11 affords the acid of general structure P-12.
Preparation 80 and Example 278.
1-Carboxy-2-[4-[[(2,4,6-trichlorophenyl)amino]carbonyl] phenyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-ethyl ester WO 99/67230 PCTIUS99/1 4233 -228- H o N OH H cl 0 cl -cl oco -ci To a cooled (0-50C) mixture of Wang polystyrene resin P-1 (Advanced Chemtech, g, ca. 1.5 mmol), N-Boc-4-iodo-L-phenylalanine P-2 (Bachem, 4.00 g, 10 mmol), and PPh 3 (1.30 g, 5.0 mmol) in THF (20 mL) was added diethyl azodicarboxylate (0.80 mL, mmol) in 4 approximately equal portions at 5 min intervals. When the orange color had discharged, the mixture was warmed to ambient temperature and stirred for 5 h. The mixture was diluted with THF (30 mL) and filtered. The resin was washed with DMF, THF and MeOH and dried in vacuo to afford the esterified resin P-3 (2.68 g) as a colorless powder: 3 C NMR (100 MHz, CD,C1 2 4 mm MAS probe) 8 171.86, 155.33, 137.85, 136.40, 131.87, 128.00, 92.74, 80.09, 54.05, 38.05, 28.51.
N, was bubbled through a mixture of N-Boc-4-iodo-L-phenylalanine functionalized Wang resin P-3 (500 mg, ca. 0.3 mmol), PPh 3 (0.21 g, 0.8 mmol), 2,4,6-trichloroaniline (0.98 g, 5.0 mmol) and DIEA (3.48 mL, 20 mmol) in NMP (20 mL) for 10 min. Pd 2 dba 3 (0.18 g, 0.2 mmol) was added and the reaction mixture was placed under a CO atmosphere and heated (bath temp. 70 for 18 h. Upon cooling to ambient temperature, the mixture was diluted with 3% sodium diethyldithiocarbamate in 95:5 NMP:DIEA (10 mL).
After an additional 10 min, the mixture was filtered and the resin washed with NMP, THF and MeOH and dried in vacuo to afford the functionalized resin P-4 as a colorless powder.
Resin P-4 was swollen with methylene chloride (0.5 mL) and diluted with 95:5
TFA:H
2 0 (10 mL). After 90 min the mixture was filtered and the resin washed with TFA (3 x 5 mL) and CH 2
CI
2 The combined filtrates were concentrated in vacuo and the residue lyophilized from glacial acetic acid to provide the amino acid P-5 (152 mg) as a powder which was used without purification. Physical data as follows: MS (FAB) m/z (rel.
intensity) 387 42), 427 426 389 387 366 279 177 146 119 23 HRMS (FAB) caled for C 16
H,
3
CLIN
2 0 3 387.0070, found 387.0084.
WO 99/67230 PCTfUS99/1 4233 -229- The amino acid P-5 was dissolved in methanolic HCI (20 mL) and stirred at room temperature for 18 h. Concentration in vacuo afforded the methyl ester P-6 which was used without purification. Physical data as follows: MS for C,,H,IsC1 3
N
2 0 m/z 400.9 To a cooled (0-5 solution of N-ethoxycarbonyl-D-thiazolidine-4-carboxylic acid P-7 (82 mg, 0.4 mmol) and HOAt (54 mg, 0.4 mmol) in CH 2
C
2 /DMF 4 mL) was added EDC (76 mg, 0.4 mmol). After stirring for 10 min, the solution was added to the amino ester P-6 described above at 0-5 OC followed by DIEA (208 RL, 1.2 mmol).
After an additional 30 min at 0-5 OC, the solution was allowed to warm to room temperature and stirred an additional hour. Volatiles were removed in vacuo and the residue partitioned between ethyl acetate and 0.1 N aqueous HC1. The organic layer was separated, washed with 0.1 N aqueous HC1, saturated aqueous NaHCO 3 brine, dried (MgSO 4 filtered and concentrated in vacuo. Purification of the residue by flash chromatography using CH 2 Cl 2 /ethyl acetate/hexanes containing 2-propanol (0.1 as eluant afforded the ester P-8 as a powder: 'H NMR (300 MHz, CDC13) 8 7.99 (1 H), 7.87 (2 7.40 (2 7.24 (2 4.92 (1 4.70 (2 4.34 (1 4.10 (2 3.74 (3 3.20 (4 1.25 (3 3 C NMR (75 MHz, CDCl 3 5 171.2, 169.7, 165.4, 154.9, 140.6, 134.3, 133.4, 132.2, 131.3, 129.7, 128.4, 127.9, 63.1, 62.7, 60.4, 53.0, 52.6, 37.6, 21.0, 14.5; MS (ESI+) for C 2 4
H
24 C1 3
N
3 0 6 S m/z 589.9 MS (ESI-) for
C
24
H
24 C1 3
N
3 0 6 S m/z 588.0 To a cooled (0-5°C solution of the ester P-8 (72 mg, 0.12 mmol) in THF (5 mL) and water (0.5 mL) was added an 0.1 N aqueous solution of NaOH (1.3 mL, 0.13 mmol) via a syringe pump over 1 h. After an additional 45 min at 0-5 0 C, the reaction mixture was diluted with ethyl acetate and acidified with 0.25 N HC1 to a pH of ca. 3. The organic layer was separated, washed with water and concentrated in vacuo. Purification of the residue by flash chromatography using methylene chloride and methanol as eluant provided a solid which was crystallized from ethyl acetate/CH 2 Cl 2 /hexanes to afford the title compound (45 mg) as colorless solid: IR (drift) 1743, 1726, 1709, 1691, 1675, 1663, 1553, 1521, 1490, 1428, 1415, 1379, 1345, 1290, 1189 'H NMR (300 MHz, DMSOd 6 8 10.27 (1 8.35 (1 7.92 2 7.81 (2 7.37 (2 4.62 (2 4.53 (1 H), WO 99/67230 PCT/US99/14233 -230- 4.29 (2 4.00 (2 3.11 (3 2.77 (1 1.12 (3 3 C NMR (75 MHz, DMSO-d 6 8 172.9, 170.0, 165.4, 154.0, 142.5, 135.5, 133.9, 133.0, 131.6, 129.8, 128.7, 128.0, 62.0, 61.7, 53.4, 37.0, 14.8; MS (FAB) m/z (rel. intensity) 574 (MH 80), 576 574 379 160 91 81 69 57 55 43 HRMS (FAB) calcd for C 23
H
22
CI
3
N
3 0 6 S 574.0373, found 574.0358.
WO 99/67230 WO 9967230PCTIUS99/1 4233 -231- Scheme Q HS R-
RQ-
2 Q-1
H
2 N 0,RQ- 3 0 0 1 Ra S RQ 0 2 Q-2 RG" t N RO- 3
H
0
I
RS R- 2
Q-
N 0
R
3 0- H0
R
01 S- R 02 z- 0 0
H
2 N 0-5 0- 00 O Ro., S- R- 0 ('NyNH<.e oKS WO 99/67230 PCT/US99/14233 -232- RQ, and RQ-2 are defined independently as Rl3 is defined as C,.
6 alkyl or C,7,1 arylalkyl. RQ.
4 is defined as oxygen or RQ.
5 is defined as a suitable protecting group for a nitrogen (such as Boc or Fmoc) or oxygen (such as t-butyldimethylsilyl) (Greene, Wuts, Protective Groups in Organic Synthesis, John Wiley and Sons, New York, 1991).
Scheme Q describes a general method for the preparation of examples of the formula Q-7. A commercially available or readily prepared sulfur containing amino acid of structure Q-1 is condensed with a suitably protected aldehyde to afford the thiazolidine-4-carboxylic acid of general formula Q-2. Standard deprotection (Greene, Wuts, Protective Groups in Organic Synthesis, John Wiley and Sons, New York, 1991) affords intermediate Q-3 which is readily cyclized to the bicycle Q-4 using 1,1 '-carbonyldiimidazole or phosgene or a suitable equivalent. For the preparation of bicycles of general structure Q-4 in which Z is CH,, see as examples: Aszodi, J.; Bonnet, Teutsch, G. Tetrahedron 1990, 46, 1579. Baldwin, Lee, V.; Schofield, C.J. Heterocycles 1992, 34, 903. Genin, Johnson, R.L. J. Am. Chem.
Soc. 1992, 114, 8778. Siddiqui, Preville, Tarazi, Warder, Eby, P.; Gorseth, Puumala, DiMaio. J. Tetrahedron Lett. 1997, 38, 8807. (e)Subasinghe, Bontems, Mclntee, Mishra, Johnson, R.L. J. Med Chem. 1993, 36, 2356. Removal of the ester protecting group affords the acid of general structure which is condensed with amino acyl derivative Q-6 under standard peptide synthesis conditions to provide Q-7 (for a review of procedures of peptide synthesis see: Bodansky, Bodansky, A. The Practice of Peptide Synthesis; Springer-Verlag: Berlin, 1984).
Mild base hydrolysis of the ester of general structure Q-7 provides acid Q-8.
Preparation 81.
(Scheme Q, Q-2: where RQ., and RQ., are equal to hydrogen, Ro 3 is ethyl, RQ- is NH, RQ., is Boc and stereochemistry is
S\
BocN H HN OEt 0 WO 99/67230 PCT/US99/14233 -233- To a suspension of D-cysteine 1.5 hydrate hydrochloride Scheme Q where RQ and RQ, are equal to hydrogen, RQ 3 is hydrogen and stereochemistry is (5 g, 27.1 mmol) in absolute ethyl alcohol (50 mL) was added triethylorthoformate (13.5 mL, 81.2 mmol) at ambient temperature. A stream of anhydrous HC1 gas was bubbled through the solution for 30 min. The stream of anhydrous HCI gas was maintained as the mixture heated to 70 0 C for 2 h. The reaction mixture was concentrated in vacuo and the resulting residue triturated with diethyl ether to afford D-cysteine ethyl ester (4.43 g) as a white solid which was used without further purification. Physical data as follows: 'H NMR (300 MHz, D 2 0) 5 4.36 (1 4.27 (2 3.12 (2 1.25 (3 MS (ESI+) for CH,,NOS m/z 150.0 To a solution of D-cysteine ethyl ester (1.89 g, 10.2 mmol) in H 2 0 (46 mL) was added potassium acetate (1.22 g, 12.4 mmol) and t-butyl N-(2-oxoethyl)carbamate (Aldrich, 2.38 g, 12.0 mmol based on 80% purity as determined by 'H NMR) in ethyl alcohol (46 mL) at ambient temperature. After 8 h, the reaction mixture was concentrated in vacuo and the residue purified by flash chromatography using methylene chloride/methanol as eluant to afford the title compound (1.9 g) as an oil: 'H NMR (300 MHz, CDCl 3 8 5.01 (1 4.75 (1 4.26 (2 3.93 (1 3.37 (2 3.12 (1 H), 2.91 (1 1.47 (9 1.32 (3 MS (ESI+) for C, 2
H
22
N
2 0 4 S m/z 291.1 MS (ESI-) for C, 2
H
22
N
2 0 4 S m/z 288.9 Preparation 82.
(Scheme Q, Q-3: where RQ., and RQ., are equal to hydrogen, R3 is ethyl, R.4 is NH, and stereochemistry is H2N,""'
S
HN
HN -OEt 0 WO 99/67230 PCT[US99/1 4233 -234- To a cooled (10-15 0 C) solution of Q-2 (Scheme Q where Ro., and Ro., are equal to hydrogen, RQ.
3 is ethyl, RQ, 4 is NH, RQ.
5 is Boc and stereochemistry is (1.9 g) in dioxane (38 mL) was added dropwise anhydrous 4 M HCI in dioxane (156 mL). The solution was allowed to warm to ambient temperature and stirred for 2 h. The reaction mixture was concentrated in vacuo and azeotroped three times with methanol which afforded the title compound (1.72 g) as a tacky yellow solid: 'H NMR (300 MHz, CDC1 3 8 8.57 (2 5.80 (1 5.25 (1 4.39 (2 4.17 (1 3.79 (4 1.37 (3 MS (ESI+) for C 7
HI
4
N
2 0 2 S m/z 191.1 MS (ESI-) for CHI 4
N
2 0 2 S 2HC1 m/z 261.0 Preparation 83.
(Scheme Q, Q-4: where RQ.- and RQ. are equal to hydrogen, R.
3 is ethyl, Z is NH, and stereochemistry is
S",
HN N OE t O OEt o o To a cooled (0-5 0 C) solution of Q-3 (Scheme Q where RQ., and RQ., are equal to hydrogen, RoQ 3 is ethyl, RQ- 4 is NH, and stereochemistry is (1.72 g, 6.54 mmol) in THF (650 mL) was added triethylamine (2.83 mL, 20.3 mmol) and 1,1'-carbonyldiimidazole (1.11 g, 6.87 mmol). After 3 d at ambient temperature, the mixture was recooled (0-5 0
C),
treated with additional 1,1'-carbonyldiimidazole (530 mg, 3.27 mmol) and allowed to warm to ambient temperature. After 18 h, the reaction mixture was concentrated in vacuo and the resulting residue was partitioned between ethyl acetate and 0.25 N HC1. The organic layer was separated, washed with brine, dried (MgSO 4 filtered and concentrated in vacuo. The diastereomeric mixture was separated by chiral chromatography [5 x 25 cm Whelk-O I, 50 mL/min 40% Isopropanol/heptane, 210 Further purification of each isolated diastereomer by flash chromatography using methylene chloride/ethyl acetate as eluant afforded the diastereomers (255 mg, 464 mg) as oils. Physical data for the faster eluting diastereomer (analytical column conditions 0.46 x 25 cm (R,R) WO 99/67230 PCT/US99/14233 -235- Whelk-O I, 0.5 mL/min 40% IPA/heptane, 210 nm) as follows: 'H NMR (300 MHz, CDCl 3 6 5.51 (1 5.15 (1 4.32 (2 3.96 (1 3.86 (1 3.53 (1 3.26 (1 H), 3.13 (1 1.34 (3 MS (ESI+) for C 8
H
12
N
2 0 3 S m/z 217.1 (M+H) MS (ESI+) for CsH, 2
N
2 0 3 S m/z 239.0 MS (ESI-) for C 8 sH 2
N
2 0 3 S m/z 215.1 Preparation 84.
(Scheme Q, Q-5: where RQ., and RQ., are equal to hydrogen, and Z is NH).
HN N O
H
OH
0 0 To a cooled (0-5 0 C) solution of the faster eluting diastereomer of general structure Q-4 (Scheme Q where RQ., and RQ., are equal to hydrogen, RQ.
3 is ethyl, Z is NH, and stereochemistry is (100 mg, 0.46 mmol) in THF (13 mL) and H 2 0 (1.5 mL) was added via syringe pump over 1 h 0.1 N NaOH (9.7 mL, 0.97 mmol). The reaction mixture was stirred for 2 h at 0°C, acidified with 1.0 N HCI (0.97 mL) and concentrated in vacuo. The resulting residue was dried over P 2 0 5 in a vacuum desiccator to afford the title compound (87 mg) as a glassy solid which was used without further purification: MS (ESI+) for
C
s
H
8
N
2 0 3 S m/z 189.0 MS (ESI+) for C 6
H
8
N
2 0 3 S m/z 211.0 MS (ESI-) for C 6
H
8
N
2 0 3 S m/z 187.0 Preparation (Scheme Q, Q-7: where RQ., and RQ., are equal to hydrogen, Z is NH, R 5 is dichlorobenzoyl)amino]phenyl, and stereochemistry of the C-terminal amino acid is WO 99/67230 PCT/US99/14233 -236- H 0 HN N N OMe 0 0 a c 0 C1
N
HI
C1 To a cooled (0-5 0 C) suspension of Q-5 (Scheme Q where RQ., and are equal to hydrogen, and Z is NH) (87 mg, 0.46 mmol) in methylene chloride (10 mL) was added O-(7-azabenzotriaol-1-yl)-N,N,TN',N'-tetramethyluronium hexafluorophosphate
(HATU)
(175 mg, 0.46 mmol), Q-6 (Scheme Q where R, is 4 2 ,6-dichlorobenzoyl)amino]phenyl, and stereochemistry is (204 mg, 0.51 mmol) and N,N-diisopropylethylamine (0.24 mL, 1.38 mmol). After 7 h, the reaction mixture was diluted with methylene chloride, washed with 0.1 N HCI and brine, dried (MgSO 4 filtered and concentrated in vacuo.
Purification of the residue by flash chromatography using methylene chloride/ethyl acetate as eluant afforded the title compound (89 mg) as a white solid: 'H NMR (300 MHz, CDCl3) 5 7.57 (2 7.31 (3 7.13 (2 4.97 (1 4.82 (1 4.47 (1 3.80 (4 3.55 (1 3.46 (1 3.34 (1 3.08 (2 MS (ESI+) for C 2 3
H
22 C1 2
N
4 0 5 S m/z 537.0 MS (ESI+) for C 23
H,
22 Cl2N 4 0 5 S m/z 558.9 (M+Na).
Preparation 86 and Example 279.
4-[(2,6-Dichlorobenzoyl)amino]-N-[[(7aS)-hexahydro-5-oxoimidazo[5, 1-b]thiazol-3-yl]carbonyl]-L-phenylalanine (Scheme Q, Q-8: where RQ., and Re., are equal to hydrogen, Z is NH, R, is dichlorobenzoyl)amino]phenyl, and stereochemistry of the C-terminal amino acid is H 0 HN NO
OH
0 0 0 C aN
HI
C,
WO 99/67230 PCT/US99/14233 -237- To a cooled (0-5 0 C) solution of Q-7 (Scheme Q where and RQ, are equal to hydrogen, Z is NH, R, is 4 -[(2,6-dichlorobenzoyl)amino]phenyl, and stereochemistry of the C-terminal amino acid is (88 mg, 0.16 mmol) in THF (5 mL) and H20 (0.5 mL) was added via syringe pump over 1 h 0.1 N NaOH (3.4 mL, 0.34 mmol). After 2 h, the reaction mixture was partitioned between ethyl acetate and 0.1 N HCI (7 mL) and diluted with H 2 0 (20 mL). The organic layer was separated, washed with H 2 0, dried (MgSO 4 filtered and concentrated in vacuo. The resulting white solid was lyophilized from glacial acetic acid to afford the title compound (22 mg) as white solid: IR (drift) 2924, 1726, 1720, 1663, 1657, 1608, 1515, 1456, 1431, 1402, 1398, 1243, 1194, 797, 780 'H NMR (300 MHz, CD30D) 8 8.05 (1 7.61 (2 7.46 (3 7.25 (2 4.93 (2 H), 4.74 (2 4.47 (1 3.77 (1 3.39 (2 3.25 (1 3.04 (2 3 C NMR (75 MHz,
CD
3 OD) 171.8, 165.2, 164.7, 138.2, 137.7, 135.1, 133.4, 132.4, 130.9, 129.4, 121.7, 66.3, 64.4, 54.5, 44.9, 37.4, 33.9; HRMS (FAB) calcd for C 22
H
20
CL
2
N
4 0sS 523.0610, found 523.0629. MS (ESI+) for C 22
H
2 oClI 2
N
4 0S m/z 523.0 MS (ESI-) for
C
22
H
20
CI
2
N
4 0 5 S m/z 521.1 Example 280.
4 2 6 -Dichlorobenzoyl)amino]-N-[[(7aR)-hexahydro-5-oxoimidazo[5,1-b]thiazol-3-yl]carbonyl]-L-phenylalanine (Scheme Q, Q-8: where RQ., and RQ., are equal to hydrogen, Z is NH, R, is dichlorobenzoyl)amino]phenyl, and stereochemistry of the C-terminal amino acid is H 0 HN
N
O OOH 0 0 K WO 99/67230 WO 9967230PCTIUS99/1 4233 -23 8- Example 280 was prepared as described in Scheme Q from the slower eluting diastereomer of general structure Q-4 (Scheme Q where and RQ., are equal to hydrogen, R-Q- is e thyl, and Z is NH). Physical data as follows: JR (drift) 3251, 3079, 1730, 1662, 1611, 1549, 1516, 1482, 1431, 1333, 1306, 1269, 1229, 1196, 792 cm'; 'H NMR (300 MHz, CD 3 OD) 8 7.59 (2 7.44 (3 7.24 (2 4.65 (1 3.81 (1 H), 3.44 (1 3.23 (2 3.08 (2 3 C NMR (75 MHz, CD 3 OD) 8 175.3, 172.22, 164.6, 155.3, 139.7, 139.1, 136.1, 134.1, 133.9, 132.3, 131.8,130.9, 122.0, 68.4, 64.6, 45,6, 44.0, 36.3; HRMS (FAB) calcd for C2 2
H
20 C1 2
N
4 0 5 S 523.0610, found 523.0629. MIS (ESI+) for C 22
H
20 C1 2
N
4 0 5 5 m/z 523.0 MIS (ESI-) for C 2 2H 20 C1 2
N
4 0 5 S m/z 521.0 Anal. Calcd for C 22
H
2 0
C
2
N
4 0 5 S 0. 13 H2O: C, 50.26; H, 3.88; N, 10.66. Found: C, 50.72; H, 3.96; N, 10. 13. Water 0.45.
Example 28 1.
4 -[(2,6-Dichlorobenzoyl)amino]-N-[(tetrahydro-5-oxo-5H-thiazolo[3 ,2-c]oxazol-3 -yl)carbonyll-L-phenylalanine (less polar diastereomer) (Scheme Q, Q-8: where RQ-, and RQ., are equal to hydrogen, Z is 0, R, is 4-[2,6dichlorobenzoyl)amino]phenyl, and stereochemnistry of the C-terminal amino acid is
OH
0 0 WO 99/67230 PCT/US99/14233 -239- Example 281 was prepared as described in Scheme Q using commercially available (t-butyldimethylsilyloxy)acetaldehyde to form the requisite thiazolidine-4-caroxylic acid.
Physical data as follows: IR (drift) 3293, 3194, 1754, 1667, 1603, 1533, 1517, 1431, 1411, 1392, 1324, 1266, 1206, 798, 781 'HNMR (300 MHz, DMSO-d6) 5 8.44 (1 7.53 (5 7.21 (2 5.00 (1 4.84 (1 4.65 (1 4.42 (2 3.08 (2 2.91 (1 "C NMR (75 MHz, CD30D) 5 169.4, 163.8, 160.6, 136.7, 136.2, 133.9, 131.9, 130.9, 129.5, 127.9, 120.4, 68.1, 64.4, 63.2, 53.4, 36.1, 33.7; MS (ESI+) for
C
22
H,
9 C1 2
N
3 0 6 S m/z 524.0 MS (ESI-) for C 2 2
H,
9 C1 2
N
3 0 6 S m/z 522.0 Anal. Calcd for C 2 2 H,,Cl 2
N
3 0 6 S 0.31 H 2 0: C, 49.86; H, 3.73; N, 7.93. Found: C, 49.61; H, 3.82; N, 7.54. Water 1.06.
Example 282.
4-[(2,6-Dichlorobenzoyl)amino]-N-[(tetrahydro-5-oxo-5H-thiazolo[3,2-c]oxazol-3-yl)carbonyl]-L-phenylalanine (more polar diastereomer) (Scheme Q, Q-8: where and are equal to hydrogen, Z is O, R, is dichlorobenzoyl)amino]phenyl, and stereochemistry of the C-terminal amino acid is I H o
OH
0 0 0 C I SClI Example 282 (diastereomer of Example 281) was prepared as described in Scheme Q. Physical data as follows: IR (drift) 3296, 1753, 1666, 1603, 1579, 1535, 1517, 1431, 1411, 1390, 1324, 1267, 1207, 798, 781 'H NMR (300 MHz, DMSO-d6) 6 8.57 (1 7.54 (5 7.20 (2 5.23 (1 4.86 (1 4.66 (1 4.45 (2 3.22 (1 3.09 (1 2.93 2 3 C NMR (75 MHz, CD 3 OD) 5 172.8, 169.7, 163.8, 160.6, 136.8, WO 99/67230 PCT/US99/14233 -240- 136.2, 133.7, 131.9, 130.9, 129.5, 127.9, 120.2, 68.1, 64.9, 63.1, 53.6, 37.5, 36.2, 34.2; MS (ESI+) for C 2 zzHCl 2
N
3 06S m/z 523.9 MS (ESI-) for C 22
H
1 9 C1 2
N
3 06S m/z 521.9 HRMS (EI) calcd for C 22 HgCL 2
N
3 06S 523.0372, found 523.0366. Anal.
Calcd for C 22 H,,Cl 2 N30OS 0.35 H20: C, 49.79; H, 3.74; N, 7.92. Found: C, 50.14; H, 4.08; N, 8.13. Water 1.19.
Example 283.
dichlorobenzoyl)amino]phenyl]ethyl]amino]carbonyl]-2-(4-pyridinyl)-3thiazolidinecarboxylic acid 3-ethyl ester o N N OH H t Example 283 was prepared as described in Scheme B using D-cysteine and 4pyridinecarboxaldehyde to from the requisite thiazolidine carboxylic acid. Physical data as follows: IR (drift) 3055, 2981, 2928, 1679, 1604, 1535, 1515, 1450, 1431, 1406, 1378, 1331, 1194, 797, 778 'HNMR (300 MHz, CD30D) 5 8.68 (2 8.28 (2 7.55 (2 7.42 (4 7.25 (2 6.28 (1 4.70 (1 4.09 (2 3.46 (1 3.16 (1 2.94 (1 2.73 (1 1.17 (3 "C NMR (75 MHz, CD 3 OD) 8 190.2, 171.1, 162.1, 153.8, 141.4, 135.3, 134.6, 131.9, 130.3, 129.4, 128.2, 126.4, 122.4, 118.5, 61.0, 51.7, 35.5, 26.6, 11.7; MS (FAB) m/z (rel. intensity) 617 (MH 99), 621 620 619 618 617 371 179 173 124 57 HRMS (FAB) calcd for
C
28
H
26 C1 2
N
4 0S 617.1028, found 617.1019. Anal. Calcd for C 28
H
2 6 C1 2
N
4 0 6 S 0.9 HCI 1.1 H 2 0: C, 50.19; H, 4.38; N, 8.36; Cl, 15.34. Found: C, 49.79; H, 4.49; N, 8.11; Cl, 15.05. Water 2.96.
WO 99/67230 WO 9967230PCT/US99/I 4233 -24 1- Example 284.
1S)-2-Amino-lI-14-[(2,6-dichlorophenyl)methoxy]phenyl]methyl] -2oxoethyl]amino]-carbonyl]-3-thiazolidinecarboxylic acid 3 I-pyrrolidinyl)ethyl] ester 0 0 Example 284 was prepared as described in Scheme C. Physical data as follows: IR (drift) 1709, 1675, 1511, 1458, 1435, 1421, 1390, 1380, 1354, 1299, 1240, 119 1179, 1114, 765 cm-i1; 'H NMR (3 00 MHz, CD 3 OD) 8 8.40 (1 7.41 (3 7.22 (2 6.99 (2 5.29 (2 4.46 (6 3.77 (2 3.50 (2 3.19 (4 2.89 (1 2.04 (1 2.12 (4 1 3 C NMR (75 MHz, CD 3 OD) 8 174.5, 171.6, 171.1, 157.9, 153.3, 136.6, 132.2, 130.6, 130.0, 129.7, 128.3, 114.5, 64.9, 63.2, 62.1, 60.9, 54.4, 53.6, 50.0, 49.0, 37.0, 36.5, 35.2, 22.6; MIS (ESI+) for C 2 7
H
32 C1 2
N
4 0 5 S m/z 595.1 HRMS (FAB) calcd for C,2 7
H
32 C1 2
N
4 0 5 S 595.1548, found 595.153 1.
Example 285.
IS)- I -Carboxy-.2-[4+[2,6dichlorophenyl)methoxylphenyl]ethyllaminolcarbonyl].3 -thiazolidinecarboxylic acid 3- (3 -tetrahydrofliranyl) ester
OH
O C1 WO 99/67230 PCT/US99/14233 -242- Example 285 was prepared as described in Scheme A. Physical data as follows: 'H NMR (300 MHz, CD 3 OD) 6 7.41 (3 7.18 (2 6.98 (2 5.28 (2 5.15 (1 H) 4.69 (2 4.44 (1 3.82 (4 "C NMR (75 MHz, DMSO-d 6 8 172.8, 157.9, 136.7, 132.2, 130.6, 130.1, 129.5, 128.3, 114.5, 76.8, 72.7, 66.5, 64.9, 62.4, 53.5, 36.3, 34.9, 32.4; MS (FAB) m/z (rel. intensity) 569 75), 571 570 569 322 161 159 89 73 71 43 HRMS (FAB) calcd for
C
2 5H 26
CL,N
2 0 7 S 569.0916, found 569.0939. Anal. Calcd for C 25
H
26 C1 2 N,0 7 S: C, 52.73; H, 4.60; N, 4.92;. Found: C, 52.41; H, 4.80; N, 4.62.
Example 286.
4 2 6 -dichlorophenyl)methoxy]phenyl]methyl]-2-oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[2-(1-piperidinyl)ethyl] ester
O
C) N No N oNH2 O- Ci
CI
Example 286 was prepared as described in Scheme C. Physical data as follows: 'H NMR (300 MHz, CD30D) 6 7.43 (3 7.22 (2 6.98 (2 5.28 (2 4.66 (2 H), 4.59 (1 4.47 (1 4.47 (2 3.05 (10 1.78 (4 1.62 (2 "C NMR (75 MHz, CDO3D) 175.9, 172.9, 159.3, 138.1, 133.6, 132.0, 131.4, 131.1,129.7, 120.1, 115.9, 66.3, 64.6, 63.7, 62.3, 61.9, 57.8, 55.8, 55.1, 51.3, 50.3, 38.5, 37.8, 36.6, 25.0, 23.5; MS (ESI+) for C 28
H
34
CI
2
N
4 :OsS m/z 609.0 WO 99/67230 PCT/US9/1 4233 -243- Example 287.
(4S)-4-[[[(1S)-2-Amino-l -[[4-[(2,6-dichlorophenyl)methoxyjphenyl]methyl]-2-oxoethyl]aminolcarbonyl]-3-thiazolidinecarboxylic acid 3-[2-(4-methyl-1 -piperazinyl)ethyl] ester H 0 Nf
NH
2 a-~o ci 0 C1
C,
Example 287 was prepared as described in Scheme C. Physical data as follows: 'H NMR (300 MHz, CD 3 OD) 6 7.41 (3 7.21 (2 6.98 (2 5.28 (2 4.67 (1 H), 4.58 (1 4.46 (1 4.23 (2 3.20 (2 2.71 (13 2.50 (3 MS (ESI±) for
C
28
H
3 ,C1 2
N,
5 0 5 m/z 624.0 Anal. Calcd for C 2
,H
3 5 C1 2 NOS 0.5 C,H 4 0 2 C, 49.82; H, 5.54; N, 10.02. Found: C, 49.82; H, 5.77; N, 9.65.
Example 288.
S)-2-Amino-l 4 2 ,6-dichlorobenzoyl)amino]phenylmethyl]-2-oxoethyl].
amino]carbonyl]-3-thiazolidinecarboxylic acid 3-[2-(4-morpholinyl)ethyl] ester H I
NH
2 0 N Na e HCI WO 99/67230 PCT/US99/14233 -244- Example 288 was prepared as described in Scheme C. Physical data as follows: IR (drift) 1671, 1603, 1536, 1518, 1430, 1415, 1361, 1349, 1324, 1269, 1194, 1134, 1118, 1104, 799 cm'; 'H NMR (300 MHz, CD 3 OD) 5 7.62 (1 7.46 (3 7.31 (2 4.68 (3 4.41 (3 3.95 (4 3.48 (5 3.22 (3 2.90 (2 "C NMR (75 MHz, CD 3
OD)
8 175.8, 173.1, 165.3, 154.7, 138.2, 137.6, 135.2, 133.3, 132.4, 130.9, 129.4, 121.7, 121.5, 65.0, 64.8, 63.7, 60.6, 57.4, 55.8, 53.6, 51.4, 50.4, 38.6, 38.1, 36.6, 35.0; MS (ESI+) for
C
2 7
H,
3 Cl1 2
N
5 0S m/z 623.9 HRMS (FAB) calcd for C 2
,H
31 ClI 2
N
5 0S+H 624.1450, found 624.1452.
WO 99/67230 PCTIUS99/14233 -245- Scheme R.
WO 99/67230 PCT/US99/14233 -246- Scheme R Scheme R teaches a general method for the preparation of Examples corresponding to structures R-4 and R-5, where RF is nitrogen. Thus reaction of the amide R-1 (obtained from the imide with the organozinc derived from a suitable protected P-iodoalanine, provides the acylamino azaphenylalanine R-2. N-deprotection of R-2 gives the aminoester R-3, that is used (as exemplified by reagent A-4 of Scheme A, and by reagent B-5 of Scheme B) for the synthesis of Examples R-4 and R-5 of this invention.
Preparation 87 (Scheme R: R-1 where R, is N, R, is H, R, is and R, is 2,6-dichloro) 2 6 -Dichloro-N-(5-iodo-2-pyridinyl)benzamide
(C
2
H
7
CI
2
IN
2 A mixture of J-2 and NH 2 NH2H 2 0 in MeOH is refluxed for 6 h under Ar. The reaction mixture is cooled, and the MeOH is removed in vacuo. The residue is partitioned between H,O and EtOAc.
The EtOAc extracts are dried, filtered and concentrated to give a brown-colored solid, that is purified by silica flash chromatography (99:1 toluene/EtOAc) to provide Preparation 87: TLC (98:2 toluene/EtOAc) Rf= 0.43; 'H NMR (CDC1 3 300 MHz) 5 10.23 8.26 7.99 7.57 7.41-7.31 1 3 C NMR (CDCl 3 75 MHz) 8 163.40, 153.22, 150.95, 147.16, 135.83, 132.72, 131.90, 130.34, 128.65, 128.50, 117.09, 86.55; MS (ESI) 393,391.
Preparation 88 (Scheme R: R-2 where R 6 is N, R, is H, R, is R, is 2,6-dichloro and the stereochemistry is S) 2 ,6-Dichlorobenzoyl)amino]-at- [[(1,1-dimethylethoxy)carbonyl]amino]-3pyridinepropanoic acid methyl ester (C 2
H
2 3 C1 2
N
3 To an amberized flask containing activated Zn dust (0.0.802 g, 12.27 mmol) under Ar is added dry THF (6 mL) and 1,2dibromoethane (0.045 mL). This suspension is brought briefly to a gentle reflux, and then is cooled to rt. A solution of TMSCl (1 M in THF, 0.39 mL) is added. The reaction mixture is stirred at 45 5 oC for 30 min, and then is cooled to rt. To this mixture is added WO 99/67230 PCT/US99/14233 -247a degassed solution of N-[(l,1-dimethylethoxy)carbonyl]-3-iodo-L-alanine methyl ester (4.04 g, 12.27 mmol) in 2:1 N,N-dimethylacetamide/THF (18 mL). The reaction mixture is stirred at 45 5 °C for 5 h, and then is cooled to 0 To this mixture is added PdCl 2 (PPh 3 2 (0.428 g) followed immediately by a degassed solution of Preparation 87 in 1:1 N,N-dimethylacetamide/THF (19 mL). This reaction mixture is stirred at 45 5 OC for 44 h. It is cooled to 0 and is quenched with cold aqueous satd NH 4 C1. This mixture is extracted with EtOAc. The combined EtOAc extracts are washed with aqueous satd NH 4
CI
and brine. The EtOAc extracts are dried, filtered and concentrated to provide a greenbrown colored oil, that is purified by silica flash chromatography (steps of 750:250:1, 700:300:1 and 650:350:1 heptane/EtOAc/iPrOH) to provide Preparation 88: TLC Rf= 0.28 (7:3 hexanes/EtOAc).
Preparation 89 (Scheme R: R-3 where R6 is N, R, is H, R s is R 9 is 2,6-dichloro and the stereochemistry is S) (S)-ac-Amino-6-[( 2 6 -dichlorobenzoyl)amino]-3-pyridinepropanoic acid methyl ester dihydrochloride salt (C, 1
H,
1 sCIN 3 03*2HCI, A solution of Preparation 88 (0.812 g, 1.73 mmol) in 4 M HC1 in dioxane (20 mL) is stirred under Ar at rt for 20 h. The reaction mixture is concentrated in vacuo, and the residue is taken up in HO (60 mL).
This aqueous mixture is extracted with Et2O (3 60 mL), and the Et20 extracts are discarded. The aqueous solution is frozen and lyophilized to provide Preparation 89: IR (diffuse reflectance) 3021, 2995, 2953, 2893, 2884, 2866, 2853, 2844, 2341, 2015, 1916, 1749, 1646, 1569, 1252 MS (EI) 367 282, 280, 262, 175, 173, 147, 145, 109, 107, 88.
WO 99/67230 PCT/US99/14233 -248- Preparation 90 Example 289 2 2 6 -Dichlorobenzoyl)amino]-5-pyridinyl]methyl]-2methoxy- 2 -oxoethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid ethyl ester
(C
23
H
24 C1 2
N
4 0 6
S)
(Scheme R: R-4 where R, is H, R, is H, R 3 is -Et, X is S, is m is 2, n is 0, o is 1, RF is N, R, is H, R 8 is R, is 2,6-dichloro and the stereochemistry is [S- Cl N NH EtO O H 0 Cl N NH S
O
To a mixture of the N-acylthiazolidinecarboxylic acid (0.292 g, 1.42 mmol), HOAt (0.193 g, 1.42 mmol) in 4:1 CHC1 2 /DMF (5.25 mL) at 0 °C is added EDC (0.272 g, 1.42 mmol). This reaction mixture is stirred at 0 OC for 20 min. Solid Preparation 89 (0.568 g, 1.29 mmol) and NMM (0.316 mL, 3.27 mmol) are added. The resulting reaction mixture is stirred at 0 OC for 4 h, and then is kept at 4 0 C for 40 h. The mixture is concentrated in vacuo, and the residue is taken up in CH 2
CI
2 The CH 2 C1 2 mixture is extracted with aq satd NaHCO 3 and H 2 0. The combined aqueous washes are back-extracted with
CH
2
CI
2 The combined CH,C1, extracts are dried, filtered and concentrated to a pale yellow-colored foam, that is purified by silica flash chromatgraphy (600:400:1 EtOAc/heptane/iPrOH) to give Preparation 90 (Example 289): IR (diffuse reflectance) 3275, 1742, 1697, 1665, 1586,1559, 1532,1476,1427,1398,1382, 1351, 1344,1311, WO 99/67230 PCT/US99/14233 -249- 1284, 1279, 1242, 1223, 1196, 1100, 1023, 802, 787, 770, 697 MS (EI) 554 495, 422, 395, 352, 315, 293, 280, 172, 160, 144, 116, 107, 88, Example 290 R *)]-4-[[[1-Carboxy-2-[2-[(2,6-dichlorobenzoyl)amino]-5pyridinyl]ethyl]amino]carbonyl]-3-thiazolidinecarboxylic acid ethyl ester
(C
22 H22CI 2
N
4 0 6 S, Example 290) (Scheme R: R-4 where R, is H, R, is H, R 3 is -Et, X is S, is m is 2, n is 0, o is 1, R is N, R 7 is H, R 8 is R, is 2,6-dichloro and the stereochemistry is [S- Cl N NH EtO 0 0 Cl N
OH
NH
J:
To a solution of Preparation 289 (0.400 g, 0.72 mmol) in 6:1 THF/H 2 0 (25.6 mL) at 0 °C under Ar is added slowly over 4 h (via syringe pump) an aqueous solution of NaOH (1 M, 7.92 mL). The reaction mixture is stirred an additional 1.5 h. The reaction mixture is partitioned between aqueous HCI and EtOAc. The aqueous solution is separated, and is extracted further with EtOAc. The combined EtOAc extracts are dried, filtered and concentrated to a beige-colored foam. This foam is taken up in 1:1 MeCN/H 2 0. This solution is frozen and lyophilized to give Example 290 as a beigecolored solid: mp 142-144 IR (diffuse reflectance) 3169, 3094, 3031, 2980, 2964, 2935, 1735, 1691, 1591, 1556, 1531, 1480, 1431, 1400, 1379, 1344, 1308, 1288, 1266, 1216, 1194, 1148, 799, 782, 772 Anal. C 48.72, H 4.29, Cl 12.26, N 9.95, S 5.62 (calcd C 48.81, H 4.10, Cl 13.10, N 10.35, S 5.92).
WO 99/67230 PCT/US99/14233 -250- Biological Assays Jurkat-Endothelial Cell Adhesion Assay: The following assay established the activity of the present compounds in inhibiting P,-mediated cell adhesion in a representative in vitro system. This assay measures the adhesive interactions of a T-cell line, Jurkat, known to express the ca4 I integrin, to endothelial monolayers in the presence of test compounds. The test compounds were added in increasing concentrations to T-cells and then the Tcell compound mixture was added to IL-1 stimulated endothelial cell monolayers.
The plates were incubated, washed and the percentage of attached cells was quantitated. The present assay directly demonstrates the cell adhesion inhibitory activity and adhesion modulatory activity of the compounds.
Human umbilical vein endothelial cells were purchased from Clonetics (San Diego, CA.) at passage number 2. The cells were grown on 0.5% porcine skin gelatin pre-coated flasks (Sigma, St. Louis MO.) in EGM-UV media (Clonetics, San Diego, CA) supplemented with 10% fetal bovine serum. Cells are refed every 2-3 days reaching confluence by day 4 to 6. The cells are monitored for factor VIII antigen and results show that at passage 12, the cells are positive for this antigen. The endothelial cells are not used following passage 6.
The T-cell line Jurkat was obtained from American Type Tissue Culture Collection (Rockville, MD) and the cells were cultured in RPMI containing 10 fetal calf serum. The cells were washed twice in Hank's Balanced Salt Solution (HBSS) and resuspended in Dulbecco's Minimal Eagle's Media (DMEM) containing 2.5 mg/ml Human Serum Albumin (HSA). Jurkat cells (x l06 cells/ml) were stained with 10 ng/ml BCECF-AM (Molecular Probes, Eugene, OR)) in HBSS without phenol red. The cells were loaded with BCECF for 60 minutes in the dark at 370C, washed 2 times, and resuspended in DMEM-HSA solution.
Confluent endothelial monolayers, grown in 96-well tissue culture plates, were stimulated for 4 hr. at 37 °C with 0.1 ng/ml (-50 U/ml) recombinant IL-1 (Amgen, Thousand Oaks, CA). Following this incubation, the monolayers were washed twice with HBSS and 0.1 ml of DMEM-HSA solution was added. Jurkat WO 99/67230 PCT/US99/14233 -251cells (5 x 10' cells) were combined with the appropriate concentration of the test compound and 0.1 ml of the Jurkat cell-compound mixture was added to the endothelial cell monolayers. Generally, 100, 20, 5 and 1.25 IM compound concentrations were tested. These concentrations are adjusted downward for analogs found or thought to be more potent. The plates were placed on ice for minutes to allow for Jurkat cell settling and the plates were incubated at 37 oC for minutes. Following this incubation, the monolayers were washed twice with PBS containing 1 mM calcium chloride and 1 mM magnesium chloride and the plates were read using a Millipore Cytofluor 2300 (Marlboro, Fluorescence in each well was measured as Arbitrary Fluorescence Units and percent adhesion in the absence of compound was adjusted to 100% and the adhesion in the presence of compound was calculated. Monolayers were also fixed in 3% paraformaldehyde and evaluated microscopically to verify the adhesion. This procedure is a modification of a previously published method (Cardarelli et al., J.
Biol. Chem. 269:18668-18673 (1994)).
Jurkat-CS-1 assay The CS-1 derived peptide, CLHPGEILDVPST, and the scrambled control peptide, CLHGPIELVSDPT, were synthesized on a Beckman 990 synthesizer using t-Boc methodology. The peptides were immobilized onto microtiter plates using the heterobifunctional crosslinker 3 -(2-pyridyldithio)propionic acid Nhydroxysuccinimide ester (SPDP) as reported by Pierschbacher et al., Proc. Natl.
Acad. USA, 80:1224-1227 (1983). Microtiter plates were coated with 20 ltg/ml HSA for 2 hr. at room temperature, washed once with PBS and derivatized with pg/ml SPDP for 1 hr. After washing, 100 tl of a 100 p.g/ml cysteine containing peptide solution which had been recently dissolved was added to the wells and allowed to crosslink to the plates overnight at 4 Unbound peptide was removed from plates by washing with PBS. To block non-reacted sites, the plates are coated with 100 pl of a 2.5 mg/ml BSA solution in PBS for 1 hr. at 37 100 p.l of Jurkat cells (2.5 x 106 cells/ml) in DMEM plus BSA (2.5 mg/ml) was mixed with WO 99/67230 PCT/US99/14233 -252an appropriate concentration of the compound to be tested and the mixture was added to peptide coated dishes and incubated for 1 hr. at 37 Generally 100, and 1.25 utM concentrations of the compound were tested. The concentrations of the compound were adjusted downward for compounds thought or found to be more potent.
Following this incubation the plates were washed once with PBS and the attached cells were fixed with 3% paraformaldehyde in PBS and stained with toluidine blue in 3.7% formaldehyde. The cells were stained overnight at room temperature and the optical density at 590 nm of toluidine blue stained cells was determined using a vertical pathway spectrophotometer to quantitate attachment (VMAX Kinetic Microplate Reader, Molecular Devices, Menlo Park, CA). This procedure is a modification of a previously published method (Cardarelli et al, J.
Biol. Chem., 269:18668-18673 (1994) and Cardarelli et al, Proc. Natl. Acad. Sci.
USA, 83:2647-2651 (1986)).
The preferred compounds are those which have low IC 5 0 values in the Jurkat EC assay or the Jurkat-CS-1 assay described above or which have at least moderate activity in both assays. All of the compounds of the present invention have an activity of less than 50 iM in the Jurkat CS-1 assay or less than 500 ltM in the Jurkat EC assay. Compounds with activity in the Jurkat CS-1 assay preferably have IC 5 0 values of less than 1 laM, more preferably less than 0.5 gM, most preferably less than or equal to 0.08 jtM. Compounds with activity in the Jurkat EC assay preferably have IC 50 values of less than 10 jiM, more preferably less than pM, most preferably less than or equal to 0.8 .M.
In the Jurkat EC Assay, IC 5 o value ranges are depicted by A, B, and C and in the Jurkat CS-1 Assay, IC 5 0 value ranges are depicted by D, E, and F. These ranges are as follows: In vitro data: EC: A 2 1 tM; 1 pM B 0.25 pM; C 0.25 pM CS-1 D 0.75 pM; 0.75 tM E 0.05 uM; F 0.05 pM.
WO 99/67230 WO 9967230PCTIUS99/I 4233 -253- EX. JKIEC JKICS-1 1 A
E
2 C
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3 C
F
4 C
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B
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6 A
D
7 A
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8 B
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9 C
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C
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36 C
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37 C
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38 A
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42 B
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WO 99/67230 WO 9967230PCTIUS99/1 4233 -254- Ex. JK/EC KC- 43 CF 44 AD B 46 AD 47 C F 48 A
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49 A
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A E 51 B
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52 C E 53 C
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54 A E C F 56 A E 57 A
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59 A
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61 B E 62 C F 63 C
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66 C E 67 A
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69 A D B
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77 A
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78 A
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79 A
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F 84 C F WO 99/67230 WO 9967230PCT[US99/1 4233 Ex. JK/EC
KC-
C 86 A 87
AD
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89 A
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91 A
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92 A
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93 A
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129 B
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131 B
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133 A
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135 B
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WO 99/67230 WO 9967230PCT[US99/1 4233 -256- Ex. JKIEC JKICS-1 137 B
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139 A
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140 A
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141 A
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142 A
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143 A
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144 A
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145 A
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146 A
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147 A
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148 C
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150 B
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156 A
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157 C
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167 B
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169 A 170
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172 C
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177 A
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178 A
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WO 99/67230 WO 9967230PCTIUS99/1 4233 -257- Ex. JKIEC JKJCS-1 181 A
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182 B
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183 184 C
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185 C
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186 C
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187 A
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188 A
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189 A
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190 B
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191 A
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192 C
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200 A
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218 219
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221 A 222 A
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WO 99/67230 WO 9967230PCTIUS99/1 4233 -258- Ex. JKIEC JKICS-1 225 A
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226 A
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227 A
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228 A
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229 A
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230 A
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231 A
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232 A
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233 A
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234 A
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235 A
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236 A
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237 A
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238 A
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239 A
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240 A
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241 A
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242 A
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243 B
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244 B
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245 C
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246 B
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247 B
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248 B
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249 B
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250 C
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251 C
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252 C
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253 C
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254 B
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255 A
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256 C
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257 B
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259 C
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260 B
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261 B
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262 C
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263 B
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265 C
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WO 99/67230 WO 9967230PCTJUS99/1 4233 -259- Ex. JKIEC JK/CS-1 266 C F 267 A E 268
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278 279 280 281
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284 285 286 287 288 290 Biological Activity in Dextran Pleurisy Model Certain compounds of the present invention were tested in a Dextrang pleurisy model.
Rationale for Developing an ctfl Integrin Antagonist to Treat Inflammatory Diseases VLA-4. a member of the 0 1 integrin family of adhesion molecules, is thought to play a critical role in several types of inflammatory disease processes by promoting leukocyte adhesion to vascular cell adhesion molecule (VCAM- 1) and the CS-lI domain of fibronectin in extracellular tissue matrix (Elices MJ, Osborn L, Takada Y, Grouse C, Luhowskyj S. Hemler M. Lobb RR. VCAM- I on activated endothelium interacts with the leukocvte inteerin VLA-4 at a site distinct from the VLA-4-fibronectin binding site. Cell; 577-584. 1990, Humphries MJ. Akivama SK. Komoriya A, Olden K. Yamada KM.
WO 99/67230 PCT/US99/14233 -260- Identification of an alternatively-spliced site in human plasma fibronectin that mediates cell type-specific adhesion. J Cell Biol; 103: 2637-2647. 1986, Wayner EA, Garcia-Pardo A, Humphries MJ. McDonald JA, Carter WG. Identification and characterization of the T lymphocyte adhesion receptor for an alternative cell attachment domain (CS-1) in plasma fibronectin. J Cell Biol; 109: 1321-1330, 1989, Guan J-L, Hynes RO. Lymphoid cells recognize an alternatively-spliced segment of fibronectin via the integrin cca~,. Cell; 53-61, 1990) Of the cell types expressing VLA-4, the major emphasis has been on eosinophils, lymphocytes, and monocytes. Validation of the role of VLA-4 has relied predominantly on the use of anti-VLA-4 antibodies which have been shown to suppress delayed-type hypersensitivity responses (Issekutz TB. Dual inhibition of VLA-4 and LFA-1 maximally inhibits cutaneous delayed-type hvpersensitivitv-induced inflammation.
Am J Pathol; 143: 1286-1293, 1993, Scheynius A. Camp RL. Pur6 E. Reduced contact sensitivity reactions in mice treated with monoclonal antibodies to leukocvte functionassociated molecule-I and intercellular adhesion molecule-1. J Immunol; 150: 655-663, 1993, Ferguson TA, Kupper TS. Antigen-independent processes in antigen-specific immunity. J Immunol; 150: 1172-1182, 1993, Chisholm PL, Williams CA, Lobb RR.
Monoclonal antibodies to the integrin a-4 subunit inhibit the murine contact hypersensitivity response. Eur J Immunol; 23: 682-688. 1993. Elices MJ, Tamraz S, Tollefson V, Vollger LW. The integrin VLA-4 mediates leukocvte recruitment to skin inflammatory sites in vivo. Clin Exp Rheumatol; 11 (Suppl 8) S77-80), 1993, experimental allergic encephalomyelitis (Yednock TA, Cannon C, Fritz LC, Sanchez- Madrid F, Steinman LM, Karin N. Prevention of experimental autoimmune encephalomyelitis by antibodies against cc integrin. Nature; 356: 63-66, 1992, Canella B, Raine CS. The VCAM-I/VLA-4 pathway is involved in chronic lesion expression in multiple sclerosis J Neuropathol Exp Neurol; 52: 311. 1993), HIV-induced encephalitis (Sasseville VG. Newman W, Brodie SJ. Hesterberg P, Pauley D, Ringler DJ.
Monocvte adhesion to endothelium in simian immunodeficiency virus-induced AIDS encephalitis is mediated by vascular cell adhesion molecule-l1/c I integrin reactions. Am J Pathol; 144: 27-40, 1994), pulmonary inflammation and airway hyperreactvity in asthma (Abraham WM, Sielczak MW. Ahmed A, Cortes A. Lauredo IT. Kim J. Pepinsky, B, et al.
WO 99/67230 PCT/US99/14233 -261- (.x-integrins mediate antigen-induced late bronchial responses and prolonged airway hvperresponsiveness in sheep. J Clin Invest: 93: 776-787. 1994. Pretolani M. Ruffid C, Roberto LapaeSilva J, Joseph D. Lobb RR. Vargaftig BB. Antibody to very late activation antigen 4 prevents antigen-induced bronchial hvperreactivitv and cellular infiltration in the guinea-pig airways. J Exp Med: 180: 795-805, 1994), experimental models of autoimmune-mediated diabetes (Yang X-D, Karin N, Tisch R, Steinman L, McDevitt HO.
Inhibition of insulitis and prevention of diabetes in non-obese diabetic mice by blocking L-selectin and very late antigen 4 adhesion receptors. Proc Natl Acad Sci USA; 10494-10498, 1993, Burkly LC, Jakubowski A, Hattori M. Protection against adoptive transfer of autoimmune diabetes medicated through very late antigen-4 integrin. Diabetes; 43: 529-534, 1994), and experimental colitis (Podolsky DK, Lobb R. King N. Benjamin CD. Pepinsky B, Sehgal P, et al. Attentuation of colitis in the cotton-top Tamarin by anti- 4 integrin monoclonal antibody. J Clin Invest: 92: 372-380, 1993). Since eosinophils represent a major component of the inflammatory cell influx in asthmatic lung tissue we developed a simple acute inflammatory model ofVLA-4 integrin-dependent eosinophil infiltration which could be used to identify VLA-4 antagonists; such compounds would be of potential value in the treatment of asthma as well as other diseases in which VLA-4 played a role.
Materials and Methods Animals, housing and viral testing: C57BL/6 mice (Jackson, Bar Harbor. ME; 6-8 weeks old, weighing 20-25g were used throughout. All mice were acclimated for at least 7-14 days after arrival and maintained under controlled temperature (20-220C) and a 12 hr daily light cycle (6.00 A.M. 6.00 Mice were housed in laminar flow racks and checked biweekly for viral infections (mouse hepatitis virus, minute virus of mice, rodent orphan parvovirus. Sendai) with kits obtained from Oreganon Teknika (Durham. NC) using established enzyme-linked immunoabsorbent assays. Mice testing positive for any of the above were omitted from the study. All mice were fed standard laboratory chow (Upjohn Lab Rodent Irradiated Mouse Chow. #5011-3, PMI Feeds, St. Louis. MO) and acidified drinking water (pH 5.0) ad libitum.
WO 99/67230 PCT/US99/14233 -262- Induction of Inflammation by Intrapleural Injection of Dextran: Intrapleural injections were made using a 27G needle cut to 3-4 mm and blunted by filing. Injections were made by inserting the needle between the mid-intercostal ribs on the right side of the thoracic cavity.
Dextran (MW 5-40x106, St Louis, MO.) was injected as a 10% solution in saline in a volume of 100 pl/mouse. Care was taken to avoid bleeding at the site of injection at which the intercostal muscles were cut to facilitate smooth insertion of the needle.
Quantitation of Pleural Inflammatory Leukocvte Responses: Pleural leukocytes were collected as follows: 4h post-induction, pleural inflammatory exudate was removed by washing with 2 x 1.0 ml Ca++/Mg free HBSS (Gibco, Grand Island, NY) containing 45 mg EDTA/100 ml HBSS, 40C. Total leukocyte counts were made by hemocytometer following erythrocyte lysis in 2% acetic acid in PBS buffer; exudate leukocyte pellets were resuspended in serum for cytospin preparations and stained (Diff Quik, Baxter Healthcare, McGraw Park, IL) for differential leukocyte counts (neutrophils, eosinophils, and mononuclear leukocytes).
The pleural cavities of mice receiving either no intrapleural injection, or saline were washed and the cells counted in the same way to estimate baseline or saline-induced pleural leukocyte counts respectively.
Administration of compounds: All drugs were dissolved in PBS and the pH adjusted to 7.5 with NaOH. Each compound was administered intravenously through the retroorbital sinus at hourly intervals (0-3h) starting from time as indicated. Mice were carefully monitored for side effects; none were noted for the series of compounds reported herein.
The following compounds were tested for their inhibitory effects on dextraninduced leukocyte infiltration: Examples 3, 8. 9, 10. 12. 16, 37, 62, 66, 67, 99, 100, 111, 113, 115, 127. 131. 141. 184, 185, 192, PBS (saline) was administered iv. as a control.
WO 99/67230 PCT/US99/14233 -263- Inhibition of eosinophil infiltration, which was suppressed by anti-alpha-4 Mab (PS/2, was used as a readout of VLA-4 antagonist activity of the compounds tested. Data for neutrophils are also reported.
Results: Dextran pleural leukocvte response. The total pleural leukocyte counts were 255x10 4 16 SEM) cells in the normal pleural cavity; of the normal pleural leukocyte population, all cells were mononuclear (a similar response was observed following intrapleural saline injection). Four hours after intrapleural injection of dextran total pleural leukocyte counts increased to 719x10 4 SEM) and comprised 36.8x10 4 SEM) eosinophils, 292x10 4 SEM) neutrophils and 391x10 4 SEM) mononuclear leukocytes.
inhibition of Eosinophil infiltration B: 20-39; C 19 Example Dose Eos 3 50 x 2 iv A 8 100 x po C 9 50 x 2 iv A 1Ox po B 12 50 x 2 iv A 16 50 x 2 iv A 37 50 x 2 iv A 62 50 x 2 iv B 66 50 x 2 iv A 67 50 x 2 iv A 99 50 x 2 iv A 100 50 x 2 iv B 111 50 x 2 iv C 113 50 x 2 iv C WO 99/67230 PCT/US99/1 4233 -264- Example 115 127 131 141 185 184 192 Dose 50 x2 iv 50 x 2 iv 50 x 2 iv 50 x 2 iv 50 x 2iv 50 x 2iv 50 x2 iv Eos
B
B
A
B
B
B

Claims (21)

1. A compound of the formula: R2 H N )m R R 3 -YY n O wherein R, may occur one to four times and each occurrence is independently hydrogen or C, 6 alkyl; 5 R, is hydrogen. pyridyl, alkyl, alkyl)-CO,-R,,, or -CO,-R in addition, R, and R, may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: /(CH2)1 S/C NRI (CH2)/ 10 together with the carbon atom to which they are attached: R 3 is hydrogen. phenyl, C, alkyl, C 3 4 alkenyl, C,, 7 arylalkyl, alkyl)-CO,- alkenyl)-CO,-R,,, alkyl)-CO-C,. alkyl, alkyl)-O-C, alkyl. alkyl)- OH. (C. 6 alkyl)-CN. adamantyl or one of the following: WO 99/67230 WO 9967230PCTJUS99/1 4233 -266-. -(CI6alky)--K 1 H 1- 6 alky)- N Z -N Z N NN [O(C 1 3 alkAl] 13 N C0 2 -R 1 OH 0 _0 N S S NN H C0 2 -RI 1 H C0 2 -R 1 I 0 NO-) HN-N -J ZCO 2 Rjj N(C 1 3 alkyl 2 in addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N 0 R 4 is NH 2 NHOH, 10 arylalkyl), or is of the formula R 5 is a formula of the following: Fis N or CH; R, is hydrogen or halogen; -267- R 1 is -NH-Y, or -CONH-; R9 may occur one to three times and is a halogen, C1,6 alkoxy, alkyl or trifluoromethyl; Rio is C,,6 alkyl. or alkyl)-OH, or hydrogen; is hydrogen or alkyl; R 12 IS alkyl or the following formula: R 1 3 -(CH 2 )r -R R3is NorCH; W is (C 1 6 alkyl); X is S; Y and Y, are independently or -(ONRo- Z is 0, CH 2 or N-R, 1; 1/is1, 2,or-3; m is 1 or2; n isO0 or 1; o isO0 or 1; pis 0or 1; qIs~orI1; ad r is0, 1, 2 or 3; with the provisos that when Y Is R, cannot be hydrogen; and when R, is equal to alkyl), C, alkYl is not equai to cert-butyl. -268-
2. The compound according to claim 1, wherein R is a formula of the following: cl R 7 C wherein R, is hydrogen or Cl.
3. The compound according to claim 2, wherein R, is hydrogen or alkyl; in addition, R, and R, may be attached to the same carbon atom and may form a 5 carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached or be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: (CH2)/ (CH together with the carbon atom to which they are attached; n is 0; m is 2; and 0 is pisO. -269-
4. The compound according to claim 3, wherein R, is hydrogen or alkyl; R 2 is hydrogen or alkyl; in addition, R, and R 2 may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached or be attached to the same atom and form a ring of 5-8 atoms of the formula: C( RI I together with the carbon atom to which they are attached; R 3 is hydrogen, alkyl, arylalkyl, 6 aikyl)-OH, 6 alkyl)-CN, adamnantyl, phenyl, or one of the following: 1 6 aly)- 1 1-6 allc>4)-- H alkyl)-C0 2 N(CH 3 2 -6a(4-N J q N Z 1I R 4 is NH., NHOH, or is of the formula -H QN and R, Iis hydrogen or CH3.
5. The compound according to claim 3, wherein R, is hydrogen; -270- R 2 is hydrogen; R 3 is (C 2 4 6 alkenyl)-CO,-R,,, alkyl)-O-C,. 3 alkyl, (C 1 6 alkyl)-C 2 R, 1 or one of the following: N N -X OH H C0 2 N02 HN-N R, is O-R, 1 R 6 is CH; is hydrogen; R7 is hydrogen; Y is and 1 is 1. 271 The compound according to claim 1, wherein said compound is N ~H0 R,-4Y N J
7. The compound according to claim 4, wherein R, is hydrogen-, -272- R 2 is hydrogen; R 3 is C,. 6 alkyl, 6 alkyl)-CO2R,,, alkenyl)-CO 2 alkyl)-O-C,. 3 alkyl, or one of the following: K -NC0 2 -RII OH 0 N02HN-N HC0 2 -RII R, is R 6 is CH; is hydrogen or alkyl; Ris hydrogen; Y is and I is 1.
8. The compound according to claim 1, wherein R, is hydrogen or C,. 3 alkyl; in PR 2 is hydrogen or C,, 4 alkyl; naddition, R, and R 2 may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms, or be attached to the same atom and form a ring of 5-8 atoms of the formula: (CH 2 )i :::.together with the carbon atom to which they are attached; R 3 is hydrogen, C,. 4 alkyl, C 7 arylalkyl, alkyl)-OH. alkyl)-C0 2 6alkyl)-CN, adamantyl, phenyl, or one of the following: -273- N 1.6 tJ 1-6 ay N N N N I I ~-T\Z N(CI- 3 akyI) 0 '~N0 2 C0 2 R 1 1 o6 OH R, is NH 2 NHOH, or is of the formula N and is hydrogen or C 14 alkyl.
9. A compound of the formula: Sr S S S wherein R, may occur one to four times and each occurrence is independently hydrogen or C, alkyl; R 2 is hydrogen, pyridyl, alkyl, (C, 4 alkvl)-C0 2 or -C0 2 -274- in addition, R, and R, may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: S/(CH2)1\ C NEtl \(CH2) together with the carbon atom to which they are attached; R 3 is hydrogen, phenyl, alkyl, C 3 alkenyl, arylalkyl, alkyl)-C0 2 R,I, (C 2 6 alkenyl)-C 2 alkyl)-CO-C, 4 alkyl, (C, 4 alkyl)-O-C, 4 alkyl, alkyl)- OH, (C, 4 alkyl)-CN, adamantyl or one of the following: -C6a OR I H 1.-6 aky-N Z (VJ _E/72 N C0 2 -Rjj N"C2-R C0 2 -RI, 1-6 ak N a [O(C 3 GC2-RI1 OH OH 0 _0 -K~-cF 3 C3aN -Ck3 N(C 1 3 aly) 2 H C0 2 H 0 2 Ri 0 NO 2 HN-N C0 2 -Rj S. a a a. a in addition, R, and R 3 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom 15 to which they are attached: Z N 0 -275- R 4 is NH,, NHOH, -O-(C 71 o arylalkyl), or is of the formula -0 R, is a formula of the following: R 6 RQ Rg R 12 R/ R is N or CH; R, is hydrogen or halogen; R, is or-CONH-; R, may occur one to three times and is a halogen, alkoxy, alkyl or trifluoromethyl; Ro is alkyl, or (C, 4 alkyl)-OH, or hydrogen; is hydrogen or C, alkyl; Rn is alkyl or the following formula: -(CH)r R,9 R, 3 is N or CH; W is (C,6 alkyl); X is S; Y and Y, are independently or Z is O, CH 2 or :1 is 1, 2, or 3; m is 1 or 2; n is 0 or 1; is 0 or 1; p is 0 or 1; q is 0 or l;and -276- r is0, 1,2 or 3; with the provisos that when Y is R, cannot be hydrogen; when R, is equal to alkyl), C, alkyl is not equal to tert-butyl; and the compound has an IC, value of less than 5uM in a Jurkat CS-I assay and/or an IC, 0 value of less than 50pM in a Jurkat EC assay. A pharmaceutical composition comprisine: a therapeutically effective amount of the compound according to any one of claims 1 to 9; and a pharmaceutically acceptable carrier or diluent.
11. A method for treating or preventing a,p, adhesion mediated conditions in a human which comprises administering to a patient an effective amount of the compound according to any one of claims 1 to 9.
12. The method according to claim 11, wherein said condition is selected from the group consisting of rheumatoid anhritis, asthma, allergy conditions. allograft rejection, psoriasis, eczema, contact dermatitis and other skin inflammatory diseases, inflammatory and immunoinflammatory conditions including ophthalmic inflammatory conditions, inflammatory bowel diseases, atherosclerosis. and ulcerative colitis. -277-
13. The method according to claim 12, wherein said condition is asthma.
14. A compound of the formula: R 2 H 0 O R, R4 N R O wherein R, may occur one to four times and each occurrence is independently hydrogen or C, 4 alkyl; R, is hydrogen, pyridyl, C, 6 alkyl, (C, 4 alkyl)-CO 2 or -CO 2 in addition, R, and R, may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: (CH91 /(CH 2 C NRi1 (CH2)/ together with the carbon atom to which they are attached; R 3 is hydrogen, phenyl, C, alkyl, C 3 alkenyl, arylalkyl, (C, 4 alkyl)-CO 2 (C2- alkenyl)-C0 2 (C, 4 alkyl)-CO-C, 4 alkyl, (C, 4 alkyl)-O-C,6 alkyl, alkyl)- OH, (C, 4 alkyl)-CN, adamantyl or one of the following: too* -278- -CC.6~kI)- ORI r ~IO N H alcyl-N Z v t0 (V N _C -C2-I -+C0 2 -Rj, I -CGCO2Rj (C -O -O 3 alkyiA]I 3 02RI OH 0 _0 F 3 N(CI- 3 afLyf H C0 2 -R 1 H C0 2 -R 1 I 0>/NO-) HN- r C02-RII *0 C C. C eq.. C in addition, R 2 and R 3 0 may combine vbith each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N.. R 4 is -O-R 1 NH 2 NHO, arylalkyl), or is of the formula -O-CH 2 -7jJ R, is a formula of the following: R 8 -R 12 R7 R6is N or CH; R 7 is hydrogen or halogen; C C C. C C CC.. C .C C C -279- R, is -NHCO-; R, may occur one to three times and is a halogen, C, 4 alkoxy, C, 4 alkyl or trifluoromethyl; Ro is CI alkyl, or (C, 4 alkyl)-OH, or hydrogen; is hydrogen or C, alkyl; R, 2 is alkyl or the following formula: Rl 3 (CH2)r R 9 R, 3 is N or CH; W is alkyl); X is S; Y is or Z is O, CH 2 or 1 is 1, 2, or 3; m is I or 2; 35 n is 0 or 1; o is 0 or 1; *0 p is 0 or 1; q is 0 or 1; and ris 0, 1,2 or 3; with the provisos that when Y is R 3 cannot be hydrogen; and when R 4 is equal to alkyl), C, alkyl is not equal to tert-butyl. *6O* *Ee* -280- The compound according to claim 14, wherein R 5 is a formula of the following: Cl wherein R, is hydrogen or Cl.
16. The compound according to claim wherein R 2 is hydrogen or C,4 alkyl; in addition, R, and R, may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached or be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: /(CH2)/ \(cH 2 together with the carbon atom to which they are attached; n is 0 m is 2; and S 10 p isO.
17. The compound according to claim 16, wherein R, is hydrogen or C,.3 alkyl; R, is hydrogen or C,4 alkyl; 5 in addition, R, and R, may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached or *be attached to the same atom and form a ring of 5-8 atoms of the formula: be attached to the same atom and form a ring of 5-8 atoms of the formula: -281- (CH 2 together with the carbon atom to which they are attached; R 3 is hydrogen, C,.6alkyl, arylalkyl, alkyl)-OH, alkyl)-C0 2 6 alkyl)-CN, adaniantyl, phenyl, or one of the following: NN -6 aky)-- II N.,N H 16akyD f3 N 1 -6 alkyl)-N z -N Z R 4 is NH- 2 NHOH, or is of the formula N and is hydrogen or CH 3 N(CH 3 2 0 0
18. The compound according to claim 16, wherein R, is hydrogen; R 2 is hydrogen; R 3 is (C 24 6 alkenyl)-C0 2 alkyl)-O-C. 3 alkyl, alkyl)-CO 2 or one of the following: -282- N H C 2 -RI 1 N OH 0~ HN-N R, is R, is CH; is hydrogen; R, is hydrogen; Y is and I s 1. C. CC C C C C C -283-
19. The compound according to claim 14, wherein said compound is o W R4 R The compound according to claim 17, wherein R, is hydrogen; R 2 is hydrogen; 5 R3 is C,.6 alkyl, alkyl)-C0 2 (C 2 -6 alkenyl)-C0 2 alkyl)-O-C. 3 alkyl, or one of the following: C0 2 -RI0 N N OH ~~KsI0 .>N02 HN-N H co 2 -R 11 R, is R 6 is CH; :*R 1 is hydrogen or C,. 6 alkyl; R 7 is hydrogen; X is S; -284- Y is and I is 1.
21. The compound according to claim 14, wherein R, is hydrogen or C 1 3 alkyl; R, is hydrogen or 4 alkyl; in addition, R, and R 2 may be attached to the same carbon atom and may form a carbocyclic ring of 5-8 atoms, or be attached to the same atom and form a ring of 5-8 atoms of the formula: (CH 2 )l\ C (CH NRII together with the carbon atom to which they are attached; R 3 is hydrogen, C,. 6 alkyl, C 7 arylalkyl, (C. 6 alkyl)-OH, (C,6 alkyl)-C0 2 6 alkyl)-CN, adamantyl, phenyl, or one of the following: r r c r ,N 1-6 akyI-K Ii 6 aky N.N N H N -N Z S NH- C0 2 RI I N(CI-. 3 akcy) 0 NO- 7 o6 OH -(CO-6 alkyl)-N Z q 43NCF3 >i H N C0 2 R, I r rr r R, is NH,, NHOH, or is of the formula -O-CH 2 -ti N and -285- R, is hydrogen or alkyl.
22. A compound of the formula: PI2 H 0 N RI- (X 4 R N! n m R 00 wherein R, may occur one to four times and each occurrence is independently hydrogen or alkyl; R, is hydrogen, pyridyl, C,. 6 alkyl, alkyl)-C0 2 -R 1 or -C0 2 in addition, R, and R 2 may be attached to the same carbon atom and form a carbocyclic ring of 5-8 atoms together with the carbon atom to which they are attached, or they may be attached to the same carbon atom and form a ring of 5-8 atoms of the formula: (CH 2 )l\ C N, together with the carbon atom to which they are attached; R. is hydrogen, phenyl, C,-6 alkyl, C 3 .6 alkenyl, arylalkyl alkyl)-C0 2 R 11 (C 26 alkenyl)-CO,-R,,, alkyl)-CO-C.6 alkyl, alkyl)-O-C.6 alkyl, alkyl)- OH, alkyl)-CN, adamantyl or one of the following: -286- -(CI-6a -1 N-N H 1-6 alkyl>-N Z (V I-N -N Z N _C -C2-RI IC0 2 -RI N 02RI, -(CI-6al KCcJ- 2 I IO(CI-3a]kA]1-3 G-C02-RI OH 0 _0 -N S H C0 2 -RI -0\ I H C0 2 -RI I 0 N 2 HN-N O O2-Rj N(CI-acy!)A r r r in addition, R 2 and R 3 0 may combine with each other at the terminal thereof to form a ring of the following formula together with the carbon atom and the nitrogen atom to which they are attached: Z N.. R4 is N 2 NHOH, arylalkyl), or is of the formula -o-CH 2 -o§n is a formula of the following: p R, is N or CH; R, is hydrogen or halogen; -287- R. is -NHCO-; R 9 may occur one to three times and is a halogen, C1.6 alkoxy, C1,6 alkyl or trifluoromethyl; Rio is C, 4 alkyl, or alkyl)-OH, or hydrogen; is hydrogen or C,.6 alkyl; is C 1 .6 alkyl or the following formula: -CH 2 )r C R RU 3 is N or CHI; W is (C, 4 6 alkyl); X is S; Y is or 0 Z isO0, CH2, or N-R, 1; I s 1, 2, or 3; m is Ior2; n is 0 or 1; o isO0 or 1; p isO0 or 1; q isO0 or 1; and 40 r is0, 1, 2or 3; with the provisos that when Y is R3 cannot be hydrogen; when R, is equal to alkyl), C, alkyl is not equal to tert-butyl; and the compound has an value of less than 5 iM in a Jurkat CS- I assay and/or an IC, value of less than 50±M in a Jurkat EC assay. -288-
23. A compound according to claim 1 substantially as hereinbefore described with reference to any of the Examples.
24. A compound according to claim 9 substantially as hereinbefore described with reference to any of the Examples. A compound according to claim 14 substantially as hereinbefore described with reference to any of the Examples.
26. A compound according to claim 22 substantially as hereinbefore described with reference to any of the Examples. 4
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