HETEROCYCLIC AMIDE COMPOUNDS AS CELL ADHESION INHIBITORS

COMPOSES D'AMIDE HETEROCYCLIQUES UTILISES EN TANT QU'INHIBITEURS DE L'ADHESION CELLULAIRE

English Abstract

Compounds of formula (I) are antagonists of VLA-4 and/or .alpha.4.beta.7, and
as such are useful in the inhibition or prevention of cell adhesion and cell-
adhesion mediated pathologies. These compounds may be formulated into
pharmaceutical compositions and are suitable for use in the treatment of
asthma, allergies, inflammation, multiple sclerosis, and other inflammatory
and autoimmune disorders.

French Abstract

Des composés de formule (I) sont des antagonistes de VLA-4 et/ou de .alpha.¿4?.beta.¿7? et sont, en tant que tels, utiles pour inhiber ou empêcher l'adhésion cellulaire et pour lutter contre des pathologies induites par l'adhésion cellulaire. Ces composés peuvent être formulés sous forme de compositions pharmaceutiques et sont adaptés pour être utilisés dans le traitement de l'asthme, des allergies, des inflammations, de la sclérose en plaques et d'autres maladies inflammatoires et auto-immunes.

Claims

WHAT IS CLAIMED IS:

1. A method for the treatment of diseases, disorders,
conditions or symptoms mediated by cell adhesion in a mammal which
comprises administering to said mammal an effective amount of a
compound Formula I:
Image
or a pharmaceutically acceptable salt thereof wherein:
R1 is 1) C1-10alkyl,
2) C2-10alkenyl,
3) C2-10alkynyl,
4) Cy,
5) Cy-C1-10alkyl,
6) Cy-C2-10alkenyl,
7) CY-C2-10alkynyl,
wherein alkyl, alkenyl, and alkynyl are optionally substituted with one to
four substituents independently selected from R a; and Cy is optionally
substituted with one to four substituents independently selected from Rb;
R2 is 1) hydrogen,
2) C1-10alkyl,
3) C2-10alkenyl,
4) C2-10alkynyl,
5) aryl,
6) aryl-C1-10a1kyl,

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7) heteroaryl,
8) heteroaryl-C1-10alkyl,
wherein alkyl, alkenyl, and alkynyl are optionally substituted with one to
four substituents independently selected from Ra; and aryl and
heteroaryl optionally substituted with one to four substituents
independently selected from R b;
R3 is 1) hydrogen,
2) C1-10 alkyl,
3) Cy, or
4) Cy-C1-10 alkyl,
wherein alkyl is optionally substituted with one to four substituents
independently selected from Ra; and Cy is optionally substituted with
one to four substituents independently selected from Rb;
R4 is 1) hydrogen,
2) C1-10alkyl,
3 C2-10alkenyl,
4) C2-10alkynyl,
5) Cy,
6) CY-C1-10alkynyl
7) Cy-C2-10alkenyl,
8) CY-C2-10alkynyl

wherein alkyl, alkenyl and alkynyl are optionally substituted with one to
four substituents selected from phenyl and R X, and Cy is optionally
substituted with one to four substituents independently selected from R y;
or
R3, R4 and the atoms to which they are attached together form a
mono- or bicyclic ring containing 0-2 additional heteroatoms selected from N,
O
and S;
R5 is 1) hydrogen,
2) C1-10alkyl,

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3) C2-10alkenyl,
4) C2-10alkynyl,
5) aryl,
6) aryl-C1-10alkyl,
7) heteroaryl,
8) heteroaryl-C1-10alkyl,
wherein alkyl, alkenyl and alkynyl are optionally substituted with one to
four substituents selected from R X, and aryl and heteroaryl are
optionally substituted with one to four substituents independently
selected from R y; or
R4, R5 and the carbon to which they are attached form a 3-7 membered
mono- or bicyclic ring containing 0-2 heteroatoms selected from N, O and
S;
R6, R7, and R8 are each independently selected from the group
consisting of
1) a group selected from R d, and
2) a group selected from R x; or
two of R6, R7, and R8 and the atom to which both are attached, or two of
R6, R7, and R8 and the two adjacent atoms to which they are attached,
together form a 5-7 membered saturated or unsaturated monocyclic ring
containing zero to three heteroatoms selected from N, O or S,
Ra is 1) Cy, or
2) a group selected from Rx;
wherein Cy is optionally subsituted with one to four substituents
independently selected from R c;
R b is 1) a group selected from R a,
2) C1-10 alkyl,
3) C2-10 alkenyl,
4) C2-10 alkynyl,

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5) aryl C1-10alkyl,
6) heteroaryl C1-10 alkyl,
wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl are optionally
substituted with a group independently selected from R c;
R c is 1) halogen,
2) NO2,
3) C(O)OR f,
4) C1-4alkyl,
5) C1-4alkoxy,
6) aryl,
7) aryl C1-4alkyl,
8) aryloxy,
9) heteroaryl,
10) NRfRg,
11) NRfC(O)Rg,
12 NRfC(O)NRfRg, or
13) CN;

R d and R e are independently selected from hydrogen, C1-10alkyl, C2-10
alkenyl, C2-10alkynyl, Cy and Cy C1-10alkyl, wherein alkyl, alkenyl,
alkynyl and Cy is optionally substituted with one to four substituents
independently selected from R c; or
R d and R e together with the atoms to which they are attached form a
heterocyclic ring of 5 to 7 members containing 0-2 additional
heteroatoms independently selected from oxygen, sulfur and nitrogen;

R f and R g are independently selected from hydrogen, C1-10alkyl, Cy and
Cy-C1-10alkyl wherein Cy is optionally substituted with C1-10alkyl; or
R f and R g together with the carbon to which they are attached form a
ring of 5 to 7 members containing 0-2 heteroatoms independently
selected from oxygen, sulfur and nitrogen;

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Rh is 1) hydrogen,
2) C1-10alkyl,
3) C2-10alkenyl,
4) C2-10alkynyl,
5) cyano,
6) aryl,
7) aryl C1-10alkyl,
8) heteroaryl,
9) heteroaryl C1-10alkyl, or
10) -SO2R i;
wherein alkyl, alkenyl, and alkynyl are optionally substituted with one to
four substituents independently selected from R a; and aryl and
heteroaryl are each optionally substituted with one to four substituents
independently selected from R b;
R i 1) C1-10alkyl,
2) C2-10alkenyl,
3) C2-10alkynyl, or
4) aryl;
wherein alkyl, alkenyl, alkynyl and aryl are each optionally substituted
with one to four substituents independently selected from R c;
Rx is 1) -OR d,
2) -NO2,
3) halogen
4) -S(O)m R d,
5) -SR d,
6) -S(O)2OR d,
7) -S(O)m NR d R e,
8) -NR d R e,
9) -O(CR f R g)n NR d R e,
10) -C(O)R d,
11) -CO2R d,


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12) -CO2(CR f R g)n CONR d R e,
13) -OC(O)R d,
14) -CN,
15) -C(O)NR d R e,
16) -NR d C(O)R e,
17) -OC(O)NR d R e,
18) -NR d C(O)OR e,
19) -NR d C(O)NR d R e,
20) -CR d(N-OR e),
21) -CF3,
22) oxo,
23) NR d C(O)NR d SO2R i,
24) NR d S(O)m R e,
25) -OS(O)2OR d, or
26) -OP(O)(OR d)2;
R y is 1) a group selected from R x,
2) C1-10 alkyl,
3) C2-10 alkenyl,
4) C2-10 alkynyl,
5) aryl C1-10alkyl,
6) heteroaryl C1-10 alkyl,
7) cycloalkyl,
8) heterocyclyl;
wherein alkyl, alkenyl, alkynyl and aryl are each optionally substituted
with one to four substituents independently selected from R x;
Cy is cycloalkyl, heterocyclyl, aryl, or heteroaryl;
m is an integer from 1 to 2;
n is an integer from 1 to 10;



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X is 1) -C(O)OR d,
2) -P(O)(OR d)(OR e)
3) -P(O)(R d)(OR e)
4) -S(O)m OR d,
5) -C(O)NR d R h, or
6) -5-tetrazolyl;
Y is ~1) -C(O)-,
2) -O-C(O)-,
3) -NR e-C(O)-,
4) -S(O)2-,
5) - P(O)(OR4) or
6) C(O)C(O);

Z and A are independently selected from -C- and -C-C-;
B is selected from the group consisting of
1) a bond,
2) -C-
3) -C-C-,
3) -C=C-,
4) a heteroatom selected from the group consisting of
nitrogen, oxygen, and sulfur; and
5) -S(O)m-.
2. A method of Claim 1 wherein in compounds of
Formula I,
Y is S(O)2;
R1 is (1) C1-10alkyl,
(2) Cy, or
(3) Cy-C1-10 alkyl;



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wherein alkyl is optionally substituted with one to two substituents
independently selected from R a, and Cy is optionally substituted with
one to four substituents independently selected from R b.
3. A method of Claim 1 wherein said cell adhesion is
mediated by VLA-4.
4. A method of Claim 1 wherein said disease is selected
from asthma, allergic rhinitis, multiple sclerosis, atherosclerosis,
inflammatory bowel disease and inflammation.
5, A compound having the formula Ia:
Image
or a pharmaceutically acceptable salt thereof, wherein
R1, R2, R3, R4, R5, R6, R7, X, B, and Z are as defined in Claim 1 with the
proviso that R6/R7 is not oxo when attached to the carbon between N and
B, and with the further proviso that when B and Z are each C, R2, R3,
R6, and R7 are each H, then R1 is other than phenyl, 4-methylphenyl
and 5-(NR d R e)naphthyl.
6. A compound of Claim 5 wherein Z is C.
7. A compound of Claim 5 wherein B is C, C=C, C-C or
S.
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8. A compound of Claim 5 wherein X is C(O)OR d.

9. A compound of Claim 5 wherein R1 is C1-10alkyl, Cy
or Cy-C1-10alkyl wherein alkyl is optionally substituted with one to two
substituents independently selected from R a, and Cy is optionally
substituted with one to four substituents independently selected from R b.
10. A compound of Claim 5 wherein R1 is aryl optionally
substituted with one to four substituents selected from R b.

11. A compound of Claim 5 wherein R5 is H and R4 is
Cy-C1-10alkyl or Cy-C1-10alkyl, wherein alkyl is optionally substituted with
one
to four substituents selected from phenyl and R x, and Cy is optionally
substituted with one to four substituents independently selected from R y;
or R4, R5 and the carbon to which they are attached together form a 3-7
membered mono- or bicyclic carbon only ring.

12. A compound of Claim 11 wherein R4 is phenyl-C1-3
alkyl, wherein phenyl is optionally substituted with one or two groups
selected from R y.

13. A compound of Claim 5 having the formula Ib:
Image

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wherein R2 is H or C1-6 alkyl, and R1, R2, R3, R4 and R5 and X are as
defined in Claim 5.

14. A compound of Claim 13 wherin X is CO2H; R1 is
aryl optionally substituted with one to four substituents selected from R b;
R2 is H; R3 H or C1-3 alkyl; R4 is phenyl-C1-3alkyl, wherein phenyl is
optionally substituted with one or two groups selected from R y; and R5 is
H.

15. A compound of Claim 5 having the formula Ic:
Image
wherein R2 is H or C1-3 alkyl; R6 is H, C1-6 alkyl, aryl, OR d, SR d,
NR d R e, or NR d C(O)R e; B is S, C=C, C or C-C; R3 is H or C1-6alkyl, R b
and R y are as defined in Claim 5.

16. A compound of Claim 15 wherein B is C and R b is
halogen, C1-10alkoxy, cyano, or trifluoromethyl.

17. A compound selected from the group consisting of
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-leucine;

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N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-arginine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-glutamic acid;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-glycine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-(1-naphthyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-.alpha.-t-butylglycine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-3-(2-thienyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-cyclohexylalanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-3-(2-naphthyl)alanine;
N-(3,3-diphenylpropanoyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(2,4-dinitrobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-3,3-diphenylalanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-proline;
N-dansyl-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-(L)-
norleucine;
N-(2-naphthalenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(4-methoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
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N-(4-phenylbenzoyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;
N-(3,4-dimethylbenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-cysteine;
N-(4-t-butylbenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(2,5-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(2-mesitylenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(p-toluenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;
N-(4-chlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(N'-acetylsulfanilyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(4-fluorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-( 1-naphthalenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(benzylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;
N-(4-nitrobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-phenylalanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-glutamine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-(4-nitrophenyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-asparagine;
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N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-methionine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-homophenylalanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(D)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-(4-fluorophenyl)alanine;
N-(3-toluenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;
N-(4-trifluoromethylbenzenesulfonyl)-1,2,3,4-
tetrahydroisoquinoline-3(S)-carbonyl-(L)-norleucine;
N-(4-n-propylbenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(4-isopropylbenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(2,6-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(4-ethylbenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(2,4-difluorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(2-cyanobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-{L)-norleucine;
N-(4-tert-amylbenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(4-chloro-3-nitrobenzenesulfonyl)-1,2,3,4-
tetrahydroisoquinoline-3(S)-carbonyl-(L)-norleucine;
N-(3-cyanobenzoyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;
N-(3,5-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;

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N-(3,4-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbony(L)-norleucine;
N-(2-trifluoromethylbenzenesulfonyl)-1,2,3,4-
tetrahydroisoquinoline-3(S)-carbonyl-(L)-norleucine;
N-(2,3-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(2,4-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(2,5-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-serine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-isoleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-tryptophan;
N-(2,1,3-benzothiadiazole-4-sulfonyl)-1,2,3,4-
tetrahydroisoquinoline-3(S)-carbonyl-(L)-tryptophan;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-3-(3-pyridyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-3-(2-naphthyl)alanine, ethyl ester;
N-acetyl-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-(L)-
norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(R)-carbonyl-(D)-norleucine;
N-propionyl-(L)-prolyl-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(4-cyanobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(benzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;


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N-(3-nitrobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(3-trifluoromethylbenzenesulfonyl)-1,2,3,4-
tetrahydroisoquinoline-3(S)-carbonyl-(L)-norleucine;
N-(2-thienylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-carbonyl-
(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-N-methylleucine;
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-
3(S)-carbonyl-(L)-citrulline;
N-(4-iodobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-norleucine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-(3-iodo)tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(3-pyridyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-glutamic acid;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-arginine;
N-(N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl)-1-amino-
cyclopentane-1-carboxylic acid;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(3,4-
dichlorophenyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-
naphthyl)alanine, ethyl ester;
N-(3,5-dichlorobenzenesulfonyl)-{L)-prolyl-(L)-3-(4-
bromophenyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(4-
nitrophenyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(4-
thiazolyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-
chlorophenyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(4-
chlorophenyl)alanine;


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N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(4-
cyanophenyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-tyrosine, O-sulfate;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3,5-diiodotyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-aspartic acid;
N-{3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-tryptophan;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-methionine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-prolyl-(L)-norleucine;
N-(3,5-di(trifluoromethyl)benzenesulfonyl)-{L)-prolyl-{L)-3-(2-
naphthyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-thiaprolyl-(L)-3-(2-
naphthyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-thiaprolyl-(L)-norleucine;
N-[4-(N'-2-toluylureido)phenylacetyl]-(L)-thiaprolyl-(L)-3-(2-
naphthyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-3-(2-
naphthyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-pipecolyl-(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-pipecolyl-(L)-norleucine,
ethyl ester;
N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-
homophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-(3-iodo)tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-3-(2-
naphthyl)alanine;
N-[4-(N'-2-toluylureido)phenylacetyl]-(L)-pipecoliny(L)-3-(2-
naphthyl)alanine;
N-[3,5-di(trifluoromethyl)benzenesulfonyl)]-(L)-pipecolyl-(L)-3-(2-
naphthyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-pipecolyl-(L)-3-(2-
naphthyl)alanine, ethyl ester;


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N-(3,4-dimethoxybenzenesulfonyl)-(L)-octahydroisoquinoline-3-
carbonyl-(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-azetidine-2-carbonyl-(L)-
norleucine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(S)-hydroxyprolyl-(L)-3-(2-
naphthyl)alanine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-4(S)-hydroxyprolyl-(L)-
norleucine;
N-(3,4-dimethoxybenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-
norleucine;
N-(3-bis(N,N-benzenesulfonyl)aminobenzenesulfonyl)-(L)-prolyl-
(L)-norleucine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(4-pyridyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-3-(2-
naphthyl)alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-4-fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-3-iodotyrosine;
N-(3-fluorobenzenesulfonyl)-(L)-thiaprolyl-(L)-3-(2-
naphthyl)alanine;
N-(3-fluorobenzenesulfonyl)-(L)-pipecolyl-(L)-3-(2-
naphthyl)alanine;
N-(3-fluorobenzenesulfonyl)-(L)-thiaprolyl-(L)-4-
fluorophenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-4-fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-4-
fluorophenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-4-
fluorophenylalanine;


-110-




N-(3-chlorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-4-
fluorophenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-tyrosine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-tyrosine;
N-(3-fluorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-tyrosine;
N-(3-chlorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-tyrosine;
N-(3-fluorobenzenesulfonyl)-(L)-pipecolyl-(L)-4-
fluorophenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-tyrosine,
O-tert-butyl ether;
N-(3-chlorobenzenesulfonyl)-{L)-4(R)-hydroxyprolyl-(L)-tyrosine,
O-tent-butyl ether;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-tyrosine
N-(3,5-dichlorobenzenesulfonyl)-(L)-3(S)-methyl-prolyl-(L)-4-
fluorophenylalanine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-
tyrosine;
N-(3-fluorobenzenesulfonyl)-{L)-3,4-dehydroprolyl-(L)-tyrosine,
O-tert-butyl ether;
N-(3-chlorobenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-tyrosine,
O-tert-butyl ether;
N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-tyrosine;
N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-tyrosine,
O-tert-butyl ether;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-tyrosine;
N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-3-iodotyrosine;
N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-3-iodotyrosine;


-111-




N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L}-3-phenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-phenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-
phenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-
phenylalanine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-3-(4-
pyridyl)alanine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-thiaprolyl-(L)-3-(4-
pyridyl)alanine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-
phenylalanine;
N-(3-trifluoromethylbenzenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3-trifluoromethylbenzenesulfonyl)-(L)-thiaprolyl-(L)-4-
fluorophenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-tyrosine,
O-phosphoric acid;
N-(3-chlorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-tyrosine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-thiaprolyl-(L)-tyrosine;
N-(N1-methyl-4-imidazolesulfonyl}-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(D)-prolyl-(D)-4-
fluorophenylalanine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-3-(4-
pyridyl)alanine;
N-(5-(5-trifluoromethyl-2-pyridylsulfonyl)-2-thiophenesulfonyl)-(L)-
prolyl-(L)-4-fluorophenylalanine;


-112-




N-(5-(N-(4-chlorobenzoyl)aminomethyl))-2-thiophenesulfonyl)-(L)-
prolyl-(L)-4-fluorophenylalanine;
N-(5-(3-(1-methyl-5-trifluoromethyl-pyrazoyl))-2-
thiophenesulfonyl)-(L)-prolyl-(L)-4-fluorophenylalanine;
N-(3-fluorobenzenesulfonyl)-2(S)-methylprolyl-(L)-O-tert-butyl-tyrosine;
N-(3-fluorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(S)-aminoprolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-4-
fluorophenylalanine;
N-(4-bromo-5-chloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-3,5-
diiodotyrosine;
N-(5-benzoylaminomethyl-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(5-benzenesulfonyl-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3-bromo-5-chloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-3,4-dehydroprolyl-(L)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-
homophenylalanine;


-113-




N-(4-benzenesulfonyl-2-thiophenesulfonyl)-(L)-prolyl-(L)-O-tert-
butyl-tyrosine;
N-(5-benzoylaminomethyl-2-thiophenesulfonyl)-(L)-prolyl-(L)-O-
tert-butyl-tyrosine;
N-(trans-2-phenyl-ethylene-sulfonyl)-(L)-prolyl-(L)-O-tert-butyl-
tyrosine;
N-(5-benzenesuifonyl-2-thiophenesulfonyl)-(L)-prolyl-(L)-O-tert-
butyl-tyrosine;
N-(3-fluorobenzenesulfonyl)-(L)-thiaprolyl-(L)-O-tert-butyl-
tyrosine;
N-(benzylsulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-cysteine, amide;
N-(1-methyl-4-imidazolylsulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(4-(N-(4-dimethylaminophenyl)diazo)-benzenesulfonyl)-(L)-
prolyl-(L)-O-tert-butyl-tyrosine;
N-(5-(4-trifluoromethylbenzenesulfonyl)-2-thiophenesulfonyl)-(L)-
prolyl-(L)-O-tert-butyl-tyrosine;
N-(3-bromobenzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(4-methylsuifonyl-benzenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(4-methoxybenzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-3-
fluorophenylalanine;
N-(5-chloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylalanine;
N-(3-chlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methylprolyl-(L)-O-tert-
butyl-tyrosine;
N-(1(R)-(+)-10-camphorsulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(1(S)-(+)-10-camphorsulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;


-114-




N-(3,4-methylenedioxy-phenylacetyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(3-chlorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-tyrosine-O-sulfate;
N-(3-chlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-tyrosine-O-sulfate;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-cysteine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-N-methyl-
isoleucine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-O-tert-
butyl-tyrosine;
N-(3-chlorobenzenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-O-tert-butyl-tyrosine;
N-(3-cyanobenzenesulfonyl)-(L)-prolyl-(L)-tyrosine;
N-benzenesulfonyl-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(4-methylsulfonylbenzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-O-
tert-butyl-tyrosine;
N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-4(R)-aminoprolyl-(L)-4-
fluorophenylalanine;
N-(9-fluorenylmethyloxycarbonyl)-(L)-prolyl-(L)-phenylalanine;
N-(benzenesulfonyl)-(L)-prolyl-(L)-phenylalanine;
N-(n-octyl-1-sulfonyl)-(L)-prolyl-(L)-phenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-5(R)-phenyl-prolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-phenyl-prolyl-(L)-4-
iodophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-1-
carbonyl-(L)-4-fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-1,3-dihydro isoindolyl-1-carbonyl-
(L)-4-fluorophenylalanine;
N-(4-(fluorescien-4-carbonylamino)benzene sulfonyl)-(L)-prolyl-
(L)-O-tert-butyl-tyrosine;


-115-



N-(3-ethoxycarbonyl-benzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(4-iodobenzenesulfonyl)-(L)-prolyl-(L)-4-benzoyl-phenylalanine;
N-(3-(4-benzophenonyl-carbonylamino)-benzenesulfonyl)-(L)-
prolyl-(L)-O-tert-butyl-tyrosine;
N-(3-(6-(biotinylamino)-n-hexanoyl)-aminobenzenesulfonyl)-(L)-
prolyl-(L)-O- tert-butyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-[3.1.0]-3-azabicyclohexane-2-
carbonyl-(L)-4-fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl))-(L)-prolyl-(L)-3-(2-
naphthyl)alanine;
N-[4-(N'-2-toluylureido)phenylacetyl-(L)-prolyl-(L)-norleucine;
N-(3,4-dimethoxybenzoyl)-(L)-prolyl-(L)-norleucine;
N-(3,4-dimethoxybenzenesulfonyl))-(L)-pipecolyl-(L)-tryptophan;
N-(4-nitrobenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-[3,5-di(trifluoromethyl)benzenesulfonyl)]-(L)-prolyl-(L)-
norleucine;
N-(3,5-dichlorobenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-(3-trifluoromethylbenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-[4-(benzoylamino)benzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-(4-methoxy-3,5-dinitrobenzenesulfonyl)-(L)-prolyl-(L)-norleucine;
N-(3-chlorobenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-(3-trifluoromethylbenzenesulfonyl))-(L)-prolyl-(L)-3-(2-
naphthyl)alanine;
N-(3-nitrobenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-(3-cyanobenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-(3,5-dichlorobenzenesulfonyl))-(L)-prolyl-(L)-tryptophan;
N-(3-methylbenzenesulfonyl))-(L)-prolyl-(L)-norleucine;
N-(3,5-dichlorobenzenesulfonyl))-(L)-3(S)-methyl-prolyl-(L)-3-(2-
naphthyl)alanine;
N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-(3-fluorobenzenesulfonyl))-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-phenylacetyl-(L)-prolyl-(L)-3-(2-naphthyl)alanine;


-116 -




N-(3-phenylpropionyl)-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-(phenylaminocarbonyl)-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-(3,5-dichlorobenzenesulfonyl))-(L)-2-methyl-prolyl-(L)-3-(2-
naphthyl)-alanine;
N-(benzenesulfonyl)-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-(4-N'-phenylureidobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-
naphthyl)alanine;
N-(3-fluorobenzenesulfonyl)-(L)-5,5-dimethyl-prolyl-(L)-3-(2-
naphthyl)alanine;
N-(4-N'-(2-toluyl)ureidobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-
naphthyl)alanine;
N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-4-iodophenylalanine;
N-(4-N'-benzylureidobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-
naphthyl)alanine;
N-(phenyloxalyl)-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-(benzylaminocarbonyl)-(L)-prolyl-(L)-3-(2-naphthyl)alanine;
N-(3-fluorobenzenesulfonyl)-{L)-2(S)-methyl-prolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-phenylalaninamide-
N-methylsulfonamide;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
iodophenylalanine;
N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-5-methylprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-3-phenylazetidinylcarbonyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-allylprolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-
phenylalanine;


-117-




N-(3-trifluoromethylbenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
nitro-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-methyl-prolyl-(L)-4-
fluorophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
cyanophenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(aminocarbonyl)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-methyl-prolyl-(L)-4-(N-t-
butoxycarbonylaminomethyl)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-3 (R)-methyl-prolyl-(L)-4-
(aminomethyl)-phenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
acetaminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2{S)-methyl-prolyl-(L)-4-
(N'-(2-toluyl)ureido)phenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(N'-(4'-fluorophenylsulfonyl)ureido)phenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(ethoxycarbonyl)aminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-(4'-
(N'-(2-toluyl)ureido)phenylacetyl)aminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-(4'-
fluorophenylsulfonyl)aminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(phenylacetyl)aminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-(4'-
fluorobenzoyl)aminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(isobutyloxycarbonyl)aminophenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
methylsulfonylaminophenylalanine;


-118-


N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(N'-(4-fluorophenyl)ureido)phenylalanine;
N-(3-trifluoromethylbenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(N-(1,1-dioxo-1,2-isothiazolidinyl)-phenylalanine;
N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
(N'-(4-(2-oxo-1-pyrrolidinyl)-phenylalanine;
N-(3,5-dichorobenzenesulfonyl)-(L)-prolyl-(L)-4-(4'-
fluorobenzoyl)phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4'-(2-
methoxybenzoyl)phenylalanine;
N-(3,5-dichorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-(4'-
fluorobenzoyl)phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(4-
fluorobenzyl)phenyl alanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-
methoxybenzyl)phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-nitrophenoxy)-
phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(4-nitrophenoxy)-
phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-(2-
nitrophenoxy)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-aminophenoxy)-
phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-
acetylaminophenoxy)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(4-
acetylaminophenoxy)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methylprolyl-(L)-4-(2-
acetylaminophenoxy)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-2-(S)-methyl-(L)-prolyl-4-(2-
cyanophenoxy)-phenylalanine;
-119-


N-(3,5-dichlorobenzenesulfonyl)-2-(S)-methyl-(L)-prolyl-4-(4-
cyanophenoxy)-phenylalanine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-methyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-benzyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-n-butyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-cyanomethyltyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(2-methoxyethyl)
-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(2-ethoxyethyl)
-tyrosine;
N-(benzenesulfonyl)-(L)-prolyl-(L)-O-(2-methoxyethyl)-tyrosine;
N-(benzenesulfonyl)-(L)-prolyl-(L)-O-(2-ethoxyethyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(1-
pyrrolidinylcarbonyl)-tyrosine;
N-(benzenesulfonyl)-(L)-prolyl-(L)-O-(1-pyrrolidinylcarbonyl)
-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(tert-butyl acetate)
-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(4-morpholinyl-
carbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(1-(2-propanonyl)-
tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(1-
pyrrolidinylcarbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(tert-
butyl acetate)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(2-
ethoxyethyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(acetic acid)-
tyrosine, methyl ester;
-120-


N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(acetic acid)-
tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(1-(2-
propanonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(1-
pyrrolidinylcarbonyl)-tyrosine, methyl ester;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(4-
morpholinyl-carbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(2-
pyrrolylcarbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(N-
phenyl-N-methylaminocarbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(N,N-
diethyl-aminocarbonyl)-tyrosine;
N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(4-
morpholinyl-carbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-O-(N,N-
diisopropyl-aminocarbonyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(benzoyl)-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-{L)-O-(cyclopentanoyl)-
tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-(5-
tetrazolyl)methyl-tyrosine;
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-N~-
benzyl-histidine;
N-benzenesulfonyl-(L)-prolyl-2-amino-2-norbornanecarboxylic acid;
N-benzenesulfonyl-(L)-prolyl-3(R)-methyl-phenylalanine;
N-benzenesulfonyl-(L)-prolyl-(L)-2,3-methano-phenylalanine;
N-benzenesulfonyl-(L)-prolyl-(D)-2,3-methano-phenylalanine; and
N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-(5-
((1H,3H)-1,3-dimethylpyrimidine-2,4-dione))-phenylalanine.
-121-


18. A method for the treatment of diseases, disorders,
conditions or symptoms mediated by cell adhesion in a mammal which
comprises administering to said mammal an effective amount of a
compound of Claim 5.

19. A method for the treatment of asthma, allergic
rhinitis, multiple sclerosis, atherosclerosis, inflammatory bowel disease
or inflammation in a mammal which comprises administering to said
mammal an effective amount of a compound of Claim 5.

20. A pharmaceutical composition which comprises a
compound of Claim 5 and a pharmaceutically acceptable carrier thereof.

-122-

Description

CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
TITLE OF THE INVENTION
HETEROCYCLIC AMIDE COMPOUNDS AS CELL ADHESION INHIBITORS
SUMMARY OF THE INVENTION
The compounds of the present invention are antagonists of
the VLA-4 integrin ("very late antigen-4"; CD49d/CD29; or x4(31) and/or
the a4~7 integrin (LPAM-1 and a4(ip), thereby blocking the binding of
VLA-4 to its various ligands, such as VCAM-1 and regions of fibronectin
and/or a4(37 to its various ligands, such as MadCAM-1, VCAM-1 and
fibronectin. Thus, these antagonists are useful in inhibiting cell
adhesion processes including cell activation, migration, proliferation
and differentiation. These antagonists are useful in the treatment,
prevention and suppression of diseases mediated by VLA-4 and/or a4(37
binding and cell adhesion and activation, such as multiple sclerosis,
asthma, allergic rhinitis, allergic conjunctivitis, inflammatory lung
diseases, rheumatoid arthritis, septic arthritis, type I diabetes, organ
transplantation, restenosis, autologous bone marrow transplantation,
inflammatory sequelae of viral infections, myocarditis, inflammatory
bowel disease including ulcerative colitis and Crohn's disease, certain
types of toxic and immune-based nephritis, contact dermal
hypersensitivity, psoriasis, tumor metastasis, and atherosclerosis.
BACKGROUND OF THE INVENTION
The present invention relates to heterocyclic amide
derivatives which are useful for the inhibition and prevention of
leukocyte adhesion and leukocyte adhesion-mediated pathologies. This
invention also relates to compositions containing such compounds and
methods of treatment using such compounds.
Many physiological processes require that cells come into
close contact with other cells and/or extracellular matrix. Such
adhesion events may be required for cell activation, migration,
proliferation and differentiation. Cell-cell and cell-matrix interactions
are mediated through several families of cell adhesion molecules
-1-


CA 02291778 1999-11-22
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(CAMs) including the selectins, integrins, cadherins and
immunoglobulins. CAMs play an essential role in both normal and
pathophysiological processes. Therefore, the targetting of specific and
relevant CAMs in certain disease conditions without interfering with
normal cellular functions is essential for an effective and safe
therapeutic agent that inhibits cell-cell and cell-matrix interactions.
The integrin superfamily is made up of structurally and
functionally related glycoproteins consisting of a and ~i heterodimeric,
transmembrane receptor molecules found in various combinations on
nearly every mammalian cell type. (for reviews see: E. C. Butcher, Cell,
~''l, 1033 (1991); T. A. Springer, Cell, 7~C, 301 (1994); D. Cox et al., "The
Pharmacology of the Integrins." Medicinal Research Rev. 14,195 (1994)
and V. W. Engleman et al., "Cell Adhesion Integrins as Pharmaceutical
Targets." in Ann. Repts. in Medicinal Chemistry, Vol. 31, J. A. Bristol,
Ed.; Acad. Press, NY, 1996, p. 191).
VLA-4 ("very late antigen-4"; CD49d/CD29; or a,4~i1) is an
integrin expressed on all leukocytes, except platelets and mature
neutrophils, including dendritic cells and macrophage-like cells and is
a key mediator of the cell-cell and cell-matrix interactions of of these cell
types (see M. E. Hemler, "VLA Proteins in the Integrin Family:
Structures, Functions, and Their Role on Leukocytes." Ann. Rev.
Immunol. $, 365 (1990)). The ligands for VLA-4 include vascular cell
adhesion molecule-1 (VCAM-1) and the CS-1 domain of fibronectin (FN).
VCAM-1 is a member of the Ig superfamily and is expressed in uivo on
endothelial cells at sites of inflammation. (See R. Lobb et al. "Vascular
Cell Adhesion Molecule 1." in Cellular and Molecular Mechanisms of
Inflammation, C. G. Cochrane and M. A. Gimbrone, Eds.; Acad. Press,
San Diego, 1993, p. 151.) VCAM-1 is produced by vascular endothelial
cells in response to pro-inflammatory cytokines (See A. J. H. Gearing
and W. Newman, "Circulating adhesion molecules in disease.",
Immunol. Today, 14, 506 (1993). The CS-1 domain is a 25 amino acid
sequence that arises by alternative splicing within a region of
fibronectin. (For a review, see R. O. Hynes "Fibronectins.", Springer-
-2-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
Velag, NY, 1990.) A role for VLA-4/CS-1 interactions in inflammatory
conditions has been proposed (see M. J. Elices, "The integrin a4~1 ~-
4) as a therapeutic target" in Cell Adhesion and Human Disease, Ciba
Found. Symp., John Wiley & Sons, NY, 1995, p. 79).
a4~i7 (also referred to as LPAM-1 and a4~3p) is an integrin
expressed on leukocytes and is a key mediator of leukocyte trafficking
and homing in the gastrointestinal tract (see C. M. Parker et al., Proc.
Natl. Acad. Sci. USA, 89, 1924 (1992)). The ligands for a4(37 include
mucosal addressing cell adhesion molecule-1 (MadCAM-1) and, upon
activation of a4(37, VCAM-1 and fibronectin (Fn). MadCAM-1 is a
member of the Ig superfamily and is expressed in vivo on endothelial
cells of gut-associated mucosal tissues of the small and large intestine
("Peyer's Patches") and lactating mammary glands. (See M. J. Briskin
et al., Nature, 3 ' , 461 (1993); A. Hamann et al., J. Immunol., 1~, 3282
(1994)). MadCAM-1 can be induced in vitro by proinflammatory stimuli
(See E. E. Sikorski et al. J. Immunol., ,~51, 5239 (1993)). MadCAM-1 is
selectively expressed at sites of lymphocyte extravasation and specifically
binds to the integrin, a4[37.
Neutralizing anti-a4 antibodies or blocking peptides that
inhibit the interaction between VLA-4 and/or a4(37 and their ligands
have proven efficacious both prophylactically and therapeutically in
several animal models of disease, including i) experimental allergic
encephalomyelitis, a model of neuronal demyelination resembling
multiple sclerosis (for example, see T. Yednock et al., "Prevention of
experimental autoimmune encephalomyelitis by antibodies against a4~i1
integrin." N ure, 356, ~ (1993) and E. Keszthelyi et al., "Evidence for a
prolonged role of a4 integrin throughout active experimental allergic
encephalomyelitis." Neurol~, 47, 1053 (1996)); ii) bronchial
hyperresponsiveness in sheep and guinea pigs as models for the various
phases of asthma (for example, see W. M. Abraham et al., "a4-Integrins
mediate antigen-induced late bronchial responses and prolonged airway
hyperresponsiveness in sheep." J. Clin. Invest. Vii, 776 (1993) and A. A.
Y. Milne and P. P. Piper, "Role of VLA-4 integrin in leucocyte
-3-


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recruitment and bronchial hyperresponsiveness in the gunea-pig." Eur.
J. Pharmacol., 282, 243 (1995)); iii) adjuvant-induced arthritis in rats as
a model of inflammatory arthritis (see C. Barbadillo et al., "Anti-VLA-4
mAb prevents adjuvant arthritis in Lewis rats." Arthr. Rheuma
(Suppl.), 36 95 (1993) and D. Seiffge, "Protective effects of monoclonal
antibody to VLA-4 on leukocyte adhesion and course of disease in
adjuvant arthritis in rats." J. Rheumatol., 'i, 12 (1996)); iv) adoptive
autoimmune diabetes in the NOD mouse (see J. L. Baron et al., "The
pathogenesis of adoptive marine autoimmune diabetes requires an
interaction between a4-integrins and vascular cell adhesion molecule-
1.", J. Clin. Invest., 9~i, 1700 (1994), A. Jakubowski et al., "Vascular cell
adhesion molecule-Ig fusion protein selectively targets activated a4-
integrin receptors in vivo: Inhibition of autoimmune diabetes in an
adoptive transfer model in nonobese diabetic mice." J. Immunol., 155,
938 (1995), and X. D. Yang et al., "Involvement of beta 7 integrin and
mucosal addressin cell adhesion molecule-1 (MadCAM-1) in the
development of diabetes in nonobese diabetic mice", Diabetes, 46, 1542
( 1997)); v) cardiac allograft survival in mice as a model of organ
transplantation (see M. Isobe et al., "Effect of anti-VCAM-1 and anti-
VLA-4 monoclonal antibodies on cardiac allograft survival and response
to soluble antigens in mice.", Tran~lant. Proc., 26, 867 (1994) and S.
Molossi et al., "Blockade of very late antigen-4 integrin binding to
fibronectin with connecting segment-1 peptide reduces accelerated
coronary arteripathy in rabbit cardiac allograf$s." J. Clin Invest., ~5,
2601 (1995)); vi) spontaneous chronic colitis in cotton-top tamarins which
resembles human ulcerative colitis, a form of inflammatory bowel
disease (see D. K. Podolsky et al., "Attenuation of colitis in the Cotton-top
tamarin by anti-a4 integrin monoclonal antibody.", J. Clin. Invest , ~2,
372 (1993)); vii) contact hypersensitivity models as a model for skin
allergic reactions (see T. A. Ferguson and T. S. Kupper, "Antigen-
independent processes in antigen-specific immunity.", J. Immunol.,
1~, 1172 (1993) and P. L. Chisholm et al., "Monoclonal antibodies to the
integrin a-4 subunit inhibit the marine contact hypersensitivity
-4-


CA 02291778 1999-11-22
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response." E~t~r. J. Immunol., Vii, 682 (1993)); viii) acute neurotoxic
nephritis (see M. S. Mulligan et al., "Requirements for leukocyte
adhesion molecules in nephrotoxic nephritis.", J. Clin. Invest., 91, 577
(1993)); ix) tumor metastasis (for examples, see M. Edward, "Integrins
and other adhesion molecules involved in melanocytic tumor
progression.", Curr. Opin. Oncol., 7 185 (1995)); x) experimental
autoimmune thyroiditis (see R. W. McMurray et al., "The role of a4
integrin and intercellular adhesion molecule-1 (ICAM-1) in murine
experimental autoimmune thyroiditis." Autoimmunitv, 23, 9 (1996); and
xi) ischemic tissue damage following arterial occlusion in rats (see F.
Squadrito et al., "Leukocyte integrin very late antigen-4/vascular cell
adhesion molecule-1 adhesion pathway in splanchnic artery occlusion
shock." Eur. J. Pharmacol., 3~1 , 153 (1996; xii) inhibition of TH2 T-cell
cytokine production including IL-4 and IL-5 by VLA-4 antibodies which
would attenuate allergic responses (J.Clinical Investigation 100, 3083
(1997). The primary mechanism of action of such antibodies appears to
be the inhibition of lymphocyte and monocyte interactions with CAMs
associated with components of the extracellular matrix, thereby limiting
leukocyte migration to extravascular sites of injury or inflammation
and/or limiting the priming and/or activation of leukocytes.
There is additional evidence supporting a possible role for
VLA-4 interactions in other diseases, including rheumatoid arthritis;
various melanomas, carcinomas, and sarcomas; inflammatory lung
disorders; acute respiratory distress syndrome CARDS); atherosclerotic
plaque formation; restenosis; uveitis and circulatory shock (for
examples, see A. A. Postigo et al., "The a4(31/VCAM-1 adhesion pathway
in physiology and disease.", Res. Immunol:, 144, ?23 (1994) and J.-X.
Gao and A. C. Issekutz, "Expression of VCAM-1 and VLA-4 dependent
T-lymphocyte adhesion to dermal fibroblasts stimulated with
proinflammatory cytokines." Immunol. ~, 375 (1996)).
At present, there is a humanized monoclonal antibody
(Antegren0 Athena Neurosciences/Elan ) against VLA-4 in clinical
development for the treatment of "flares" associated with multiple
_5_


CA 02291778 1999-11-22
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sclerosis and a humanized monoclonal antibody (ACT-1~lLDP-02
LeukoSite) against a4(37 in clinical development for the treatment of
inflammatory bowel disease. Several peptidyl antagonists of VLA-4 have
been described (D. Y. Jackson et al., "Potent a4(31 peptide antagonists as
potential anti-inflammatory agents", J. Med. Chem., 40, 3359 (1997); H.
N. Shroff et al., "Small peptide inhibitors of a4(37 mediated MadCAM-1
adhesion to lymphocytes", Bioorg. Med. Chem. Lett., ~, 2495 (1996); US
5,510,332, W097/03094, W097/02289, W096/40781, W096l22966,
W096/20216, W096/01644, W096/06108, W095/15973). There is one report
of nonpeptidyl inhibitors of the ligands for a4-integrins (W096/31206).
There still remains a need for low molecular weight, specific inhibitors
of VLA-4- and a4~i7-dependent cell adhesion that have improved
pharmacokinetic and pharmacodynamic properties such as oral
bioavailability and significant duration of action. Such compounds
would prove to be useful for the treatment, prevention or suppression of
various pathologies mediated by VLA-4 and a4(37 binding and cell
adhesion and activation.
DETAILED DESCRIPTION OF THE INVENTION
One aspect of the present invention provides a method for
the treatment of diseases, disorders, conditions or symptoms mediated
by cell adhesion in a mammal which comprises administering to said
mammal an effective amount of a compound Formula I:
R~
R6 ,B -I ~ Ra R3
Aw ~N~X
/Y R20 R4 Rs
R'
I
or a pharmaceutically acceptable salt thereof wherein:
-6-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
R1 is 1) C1_l0alkyl,
2) C2_l0alkenyl,
3) C2_l0alkynyl,
4) Cy,
5) Cy-C1-lOalkYl,
6) Cy-C2_l0alkenyl,
) CY-C2_10~k3'nyl,
wherein alkyl, alkenyl, and alkynyl are optionally substituted with one to
four substituents independently selected from Ra; and Cy is optionally
substituted with one to four substituents independently selected from Rb;
R2 is 1) hydrogen,
2) C 1_ l0alkyl,


3) C2-l0alkenyl,


4) C2-l0~kynYl,


5) aryl,


6) aryl-C1_10a1kY1,


7) heteroaryl,
8) heteroaryl-C1_l0alkyl,
wherein alkyl, alkenyl, and alkynyl are optionally substituted with one to
four substituents independently selected from Ra; and aryl and
heteroaryl optionally substituted with one to four substituents
independently selected from Rb;
R3 is 1) hydrogen,
2) C1-10 ~Yl,
3) Cy, or
Cy-C1-1o ~kyl,
wherein alkyl is optionally substituted with one to four substituents
independently selected from Ra; and Cy is optionally substituted with
one to four substituents independently selected from Rb;
R4 is 1) hydrogen,
_7_


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
r
2) C1-10a1kYl,
3 C2_l0alkenyl,
4) C2_l0alkynyl,
5) Cy,
s) Cy-C1_lO~kYl,
?) Cy-C2_l0alkenyl,
8) CY-C2-lO~kYnYl~
wherein alkyl, alkenyl and alkynyl are optionally substituted with one to
four substituents selected from phenyl and RX, and Cy is optionally
substituted with one to four substituents independently selected from RY;
or
R3, R4 and the atoms to which they are attached together form a mono-
or bicyclic ring containing 0-2 additional heteroatoms selected from N, O
and S;
R5 is 1) hydrogen,
2) C1_lOalkyl,
3) C2-l0alkenyl,
4) C2_l0alkynyl,
5) aryl,
6) aryl-C1_l0alkyl,
?) heteroaryl,
8) heteroaryl-C1_l0alkyl,
wherein alkyl, alkenyl and alkynyl are optionally substituted with one to
four substituents selected from RX, and aryl and heteroaryl are
optionally substituted with one to four substituents independently
selected from RY; or
R4, R5 and the carbon to which they are attached form a 3-7 membered
mono- or bicyclic ring containing 0-2 heteroatoms selected from N, O and
S;
_g_


CA 02291778 1999-11-22
WO 98/53814 PCTJUS98/10940
R6, R7, and R8 are each independently selected from the group
consisting of
1 ) a group selected from Rd , and
2) a group selected from Rx; or
two of Rs, R7, and R8 and the atom to which both are attached, or two of
Rs, R7, and R8 and the two adjacent atoms to which they are attached,
together form a 5-7 membered saturated or unsaturated monocyclic ring
containing zero to three heteroatoms selected from N, O or S,
Ra is 1) Cy, or
2) a group selected from Rx;
wherein Cy is optionally subsituted with one to four substituents
independently selected from Rc;
Rb is 1) a group selected from
Ra,


2) C 1-10 ~Yh


3) C2_10 alkenyl,


4) C2_10 alkynyl,


5) aryl C1-l0alkyl,


6) heteroaryl C1_10 alkyl,


wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl are optionally
substituted with a group independently selected from Rc;
Rc is 1) halogen,


2) N02,


3) C(O)ORf


4) C1_4alkyl,


5) C 1_4alkoxy,


6) aryl,


7) aryl C1_4alkyl,


8) aryloxy,
9) heteroaryl,
10) NRfRg,
-9-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
11) NRfC(O)Rg,
12 NRfIC(O)NRfRg, or
13) CN;
Rd and Re are independently selected from hydrogen, C1_IOalkyl, C2_10
alkenyl, C2_l0alkynyl, Cy and Cy C1_l0alkyl, wherein alkyl, alkenyl,
alkynyl and Cy is optionally substituted with one to four substituents
independently selected from Rc; or
Rd and Re together with the atoms to which they are attached form a
heterocyclic ring of 5 to 7 members containing 0_2 additional
heteroatoms independently selected from oxygen, sulfur and nitrogen;
Rf and Rg are independently selected from hydrogen, C1-lO~kYl, Cy and
Cy-C1_l0alkyl wherein Cy is optionally substituted with C1-l0alkyl; or
Rf and Rg together with the carbon to which they are attached form a
ring of 5 to 7 members containing 0-2 heteroatoms independently
selected from oxygen, sulfur and nitrogen;
Rh is 1) hydrogen,
2) C1_l0alkyl,
3) C2-10a1kenyl,
4) C2-10a1kYnYl,
5) cyano,
6) aryl,
7) aryl C1_l0aikyl,
8) heteroaryl,
9) heteroaryl C1_Zpalkyl, or
10) -S02R1;
wherein alkyl, alkenyl, and alkynyl are optionally substituted with one to
four substituents independently selected from Ra; and aryl and
heteroaryl are each optionally substituted with one to four substituents
independently selected from Rb;
-10-


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Rl 1) C1_l0alkyl,
2) C2-l0alkenyl,
3) C2-l0~kynYl, or
4) aryl;
wherein alkyl, alkenyl, alkynyl and aryl are each optionally substituted
with one to four substituents independently selected from Rc;
Rx is 1) -ORd,


2) -N02,


3) halogen


4) -S(O)mRd


,
5) -SRd,


6) -S(O)20Rd


,
7) -S(O)mNRdRe


,
8) -NRdRe


,
9) -O(CRfRg)nNRdRe,


10) -C(O)Rd


11) -C02Rd,


12) -C02(CR~)nCONRdRe,


13) -OC(O)Rd


,


14) -CN,


15) -C(O)NRdRe


,
16) -NRdC(O)Re


,
17) -OC(O)NRdRe


,
18) -NRdC(O)ORe


,
19) -NRdC(O)NRdRe


,
20) -CRd(N-ORe)


,
21) -CF3,


22) oxo,


23) NRdC(O)NRd S02Ri,


24) NRdS(O)mRe,


25) -OS(O)20Rd, or


26) -OP(O)(ORd)2;


-11-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
RY is 1) a group selected from Rx,


2) C 1-10 alkyl,


) C2-10 ~kenyl,


4) C2_10 alkynyl,


5) aryl C 1-l0alkyl,


6) heteroaryl C 1-10 alkyl,


7) cycloalkyl,


8) heterocyclyl;


wherein alkyl,
alkenyl, alkynyl
and aryl are
each optionally
substituted


with one to four
substituents
independently
selected from
RX;



Cy is cycloalkyl, heterocyclyl, aryl, or heteroaryl;
m is an integer from 1 to 2;
n is an integer from 1 to 10;
X is 1) -C(O)ORd,
2) -P(O)(ORd)(ORe)


3) -P(O)(Rd)(ORe)


4) -S(O)mORd,


5) -C(O)NRdRh,
or


fi) -5-tetrazolyl;


Y is 1) -C(O)-,


2) -O-C(O)-,


3) -NRe-C(O)-,


-S(O)2-


5) - P(O)(OR4)
or


6) C(O)C(O);


Z and A are independently selected from -C- and -C-C-;
-12-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
B is selected from the group consisting of
1) a bond,
2) -C-
3) -C-C-,
3) -C=C-,
4) a heteroatom selected from the group consisting of
nitrogen, oxygen, and sulfur; and
5) -S(O)m-.
In one embodiment of the method compounds of Formula I
are those wherein Y is S(O)2 and Rl is C1-lO~kYl, Cy or Cy-C1_10 alkyl
wherein alkyl is optionally substituted with one to two substituents
independently selected from Ra, and Cy is optionally substituted with
one to four substituents independently selected from Rb.
In another embodiment of the method compounds of
Formula I are those of formula Ia, Ib or Ic.
In another embodiment, the cell adhesion is mediated by
VLA-4.
Another aspect of the present invention provides novel
compounds of Formula Ia:
R~
Rs~ ~_Z Rs
N~X
N
O.~S\\R2 O R4 Rs
R/ O
Ia
or a pharmaceutically acceptable salt thereof, wherein the variables are
as defined under formula I with the proviso that R6/R,7 is not oxo when
attached to the carbon between N and B, and with the further proviso
-13-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
that when B and Z are each C, R2, R3, R6, and R7 are each H, then Rl is
other than phenyl, 4-methylphenyl and 5-(NRdRe)naphthyl.
In one subset of Formula Ia are compounds wherein Z is C.
In another subset of Formua Ia are compounds wherein B
is C, C=C, C-C or S. Preferably B is C or C=C.
In another subset of Formula Ia are compounds wherein X
is C(O)ORd.
In another subset of Formula Ia are compounds wherein
Rl is C1-l0alkyl, Cy or Cy-C1_l0alkyl wherein alkyl is optionally
substituted with one to two substituents independently selected from Ra,
and Cy is optionally substituted with one to four substituents
independently selected from Rb. For the purpose of Rl Cy is preferably
aryl optionally substituted with one to four substituents selected from Rb.
More preferred R1 is phenyl with a substituent on the 3-position and
optionally a second substituent; the more preferred substituents are
selected from C 1_ l0alkoxy, halogen, cyano, and trifluoromethyl.
In another subset of Formula Ia are compounds wherein
R2 is H or C1_galkyl. Preferred R2 is H or C1_galkyl, more preferably H
or methyl.
In another subset of Formula Ia are compounds wherein
R3 is H or C1_galkyl. Preferred R3 is H or C1_3alkyl, more preferably H
or methyl.
In another subset of Formula Ia are compounds wherein
R5 is H and R4 is C1_l0alkyl or Cy-C1_l0alkyl, wherein alkyl is optionally
substituted with one to four substituents selected from phenyl and Rx,
and Cy is optionally substituted with one to four substituents
independently selected from RY; or R4, R5 and the carbon to which they
are attached together form a 3-7 membered mono- or bicyclic carbon only
ring. For the purpose of R4, Cy is preferably aryl, more preferably
phenyl. In a preferred embodiment, R4 is phenyl-C1_3alkyl, wherein
phenyl is optionally substituted with one or two groups selected from RY.
In one embodiment of compounds of formula Ia are
compounds of formula Ib:
-14-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
R2
O,. N O
S,.
R~ O N_ Rs
R4~
R5 X
Ib
wherein R2 is H or C1_g alkyl, and R1, R3, R4 and R5 are as defined
previously under Formula I. In a preferred embodiment X is C02H; R1
is aryl optionally substituted with one to four substituents selected from
Rb; R2 is H; R3 is H or C1_g alkyl; R4 is phenyl-C1_3alkyl, wherein
phenyl is optionally substituted with one or two groups selected from RY;
and R5 is H.
Another embodiment of compounds of Formula Ia are
compounds of the formula Ic:
B R2 R3
Rs '' N C02H
O, N
0 O ~ 1-2
~Rb)o-~ / ~ (Ry)o-2
b
R
Ic
wherein R2 is H or C 1_g alkyl, Rs is H, C 1_6 alkyl, aryl, ORd, SRd,
NRdRe, or NRdC(O)Re, B is S, C=C, C or C-C, R3 is H or C1_galkyl, Rb
and RY are as defined under Formula I. Preferably B is C and Rb is
halogen, C1_l0alkoxy, cyano, or trifluoromethyl.
-15-


CA 02291778 1999-11-22
WO 98/53814 PCT/I3S98/10940
The present compounds are generally composed of three
domains: 1) an acyl (including suifonyl) moiety, 2) a cyclic amino acid 1,
and 3) amino acid 2, and are named in a mariner similar to that used to
name oliogopeptides. Representative names used herein and their
corresponding structures are shown below (without the stereochemistry)
to illustrate the nomenclature used in the application.
N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-tetrahydroisoquinoline-3(S)-
carbonyl-(L)-leucine
CH30 / ' N
H
N
CH30 ' p ~O O C02H
N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-homophenylalanine
CI
/ ~ ,N
,S.~O N C02H
CI O O
N-(3-fluorobenzenesulfonyl)-(L)-4(R)-hydroxyprolyl-(L)-tyrosine, O-tert-
butyl ether
-16-


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HO
H
N C02H
S-O o \
I \ ~o I ~
0
F
N-(4-(N'-2-toluylureido)phenylacetyl-(L)-prolyl-(L)-norleucine
CH3 (~--~ N ~-C02H
\ N N / 1 N O (CH2)s
I ~ -,~ O \CHs
/ O
"Alkyl", as well as other groups having the prefix "alk",
such as alkoxy, alkanoyl, means carbon chains which may be linear or
branched or combinations thereof. Examples of alkyl groups include
methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl,
heptyl, octyl, nonyl, and the like.
"Alkenyl" means carbon chains which contain at least one
carbon-carbon double bond, and which may be linear or branched or
combinations thereof. Examples of alkenyl include vinyl, allyl,
isopropenyl, pentenyl, hexenyl, heptenyl, 1-propenyl, 2-butenyl, 2-
methyl-2-butenyl, and the like.
"Alkynyl" means carbon chains which contain at least one
carbon-carbon triple bond, and which may be linear or branched or
combinations thereof. Examples of alkynyl include ethynyl, propargyl,
3-methyl-1-pentynyl, 2-heptynyl and the like.
"Cycloalkyl" means mono- or bicyclic saturated carbocyciic
rings, each of which having from 3 to 10 carbon atoms. The term also
includes monocyclic rings fused to an aryl group in which the point of
attachment is on the non-aromatic portion. Examples of cycloaikyl
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include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl,
tetrahydronaphthyl, decahydronaphthyl, indanyl, and the like.
"Aryl" means mono- or bicyclic aromatic rings containing
only carbon atoms. The term also includes aryl group fused to a
monocyclic cycloalkyl or monocyclic heterocyclyl group in which the
point of attachment is on the aromatic portion. Examples of aryl include
phenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-
dihydrobenzofuranyl, benzopyranyl, 1,4-benzodioxanyl, and the like.
"Heteroaryl" means a mono- or bicyclic aromatic ring
containing at least one heteroatom selected from N, O and S, with each
ring containing 5 to 6 atoms. Examples of heteroaryl include pyrrolyl,
isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl,
thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl,
triazinyl,
thienyl, pyrimidyl, pyridazinyl, pyrazinyl, benzoxazolyl, benzothiazolyl,
benzimidazolyl, benzofuranyl, benzothiophenyl, furo(2,3-b)pyridyl,
quinolyl, indolyl, isoquinolyl, and the like.
"Heterocyclyl" means mono- or bicyclic saturated rings
containing at least one heteroatom selected from N, S and O, each of said
ring having from 3 to 10 atoms in which the point of attachment may be
carbon or nitrogen. The term also includes monocyclic heterocycle fused
to an aryl or heteroaryl group in which the point of attachment is on the
non-aromatic portion. Examples of "heterocyclyl" include pyrrolidinyl,
piperidinyl, piperazinyl, imidazolidinyl, 2,3-dihydrofuro(2,3-b)pyridyl,
benzoxazinyl, tetrahydrohydroquinolinyl, tetrahydroisoquinolinyl,
dihydroindolyl, and the like. The term also includes partially
unsaturated monocyclic rings that are not aromatic, such as 2- or 4-
pyridones attached through the nitrogen or N-substituted-(1H,3H)-
pyrimidine-2,4-diones (N-substituted uracils).
"Halogen" includes fluorine, chlorine, bromine and iodine.
Optical Isomers - Diastereomers - Geometric Isomers - Tautomers
Compounds of Formula I contain one or more asymmetric
centers and can thus occur as racemates and racemic mixtures, single
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enantiomers, diastereomeric mixtures and individual diastereomers.
The present invention is meant to comprehend all such isomeric forms
of the compounds of Formula I.
Some of the compounds described herein contain olefinic
double bonds, and unless specified otherwise, are meant to include both
E and Z geometric isomers.
Some of the compounds described herein may exist with
different points of attachment of hydrogen, referred to as tautomers.
Such an example may be a ketone and its enol form known as keto-enol
tautomers. The individual tautomers as well as mixture thereof are
encompassed with compounds of Formula I.
Compounds of the Formula I may be separated into
diastereoisomeric pairs of enantiomers by, for example, fractional
crystallization from a suitable solvent, for example methanol or ethyl
acetate or a mixture thereof. The pair of enantiomers thus obtained may
be separated into individual stereoisomers by conventional means, for
example by the use of an optically active acid as a resolving agent.
Alternatively, any enantiomer of a compound of the general
Formula I or Ia may be obtained by stereospecific synthesis using
optically pure starting materials or reagents of known configuration.
Salts
The term "pharmaceutically acceptable salts" refers to salts
prepared from pharmaceutically acceptable non-toxic bases or acids
including inorganic or organic bases and inorganic or organic acids.
Salts derived from inorganic bases include aluminum, ammonium,
calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts,
manganous, potassium, sodium, zinc, and the like. Particularly
preferred are the ammonium, calcium, magnesium, potassium, and
sodium salts. Salts derived from pharmaceutically acceptable organic
non-toxic bases include salts of primary, secondary, and tertiary
amines, substituted amines including naturally occurring substituted
amines, cyclic amines, and basic ion exchange resins, such as arginine,
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betaine, caffeine, choline, N,N~-dibenzylethylenediamine, diethylamine,
2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,
ethylenediamine, N-ethyl-morpholine, N-ethylpiperidine, glucamine,
glucosamine, histidine, hydrabamine, isopropylamine, lysine,
methylglucamine, morpholine, piperazine, piperidine, polyamine
resins, procaine, purines, theobromine, triethylamine, trimethylamine,
tripropylamine, tromethamine, and the like.
When the compound of the present invention is basic, salts
may be prepared from pharmaceutically acceptable non-toxic acids,
including inorganic and organic acids. Such acids include acetic,
benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic,
fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic,
lactic, malefic, malic, mandelic, methanesulfonic, mucic, nitric,
pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-
toluenesulfonic acid, and the like. Particularly preferred are citric,
hydrobromic, hydrochloric, malefic, phosphoric, sulfuric, and tartaric
acids.
It will be understood that, as used herein, references to the
compounds of Formula I are meant to also include the pharmaceutically
acceptable salts.
Utilities
The ability of the compounds of Formula I to antagonize the
actions of VLA-4 and/or a4~i? integrin makes them useful for preventing
or reversing the symptoms, disorders or diseases induced by the binding
of VLA-4 and or a4[37to their various respective ligands. Thus, these
antagonists will inhibit cell adhesion processes including cell activation,
migration, proliferation and differentiation. Accordingly, another
aspect of the present invention provides a method for the treatment
(including prevention, alleviation, amelioration or suppression) of
diseases or disorders or symptoms mediated by VLA-4 and/or a4~i7
binding and cell adhesion and activation, which comprises
administering to a mammal an effective amount of a compound of
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Formula I. Such diseases, disorders, conditions or symptoms are for
example (1) multiple sclerosis, (2) asthma, (3) allergic rhinitis, (4)
allergic conjunctivitis, (5) inflammatory lung diseases, (6) rheumatoid
arthritis, (?) septic arthritis, (8) type I diabetes, (9) organ
transplantation
rejection, (10) restenosis, (11) autologous bone marrow transplantation,
(12) inflammatory sequelae of viral infections, (13) myocarditis, (14)
inflammatory bowel disease including ulcerative colitis and Crohn's
disease, (15) certain types of toxic and immune-based nephritis, (16)
contact dermal hypersensitivity, (17) psoriasis, (18) tumor metastasis,
and (19) atherosclerosis.
Dose Ranges
The magnitude of prophylactic or therapeutic dose of a
compound of Formula I will, of course, vary with the nature of the
severity of the condition to be treated and with the particular compound
of Formula I and its route of administration. It will also vary according
to the age, weight and response of the individual patient. In general, the
daily dose range lie within the range of from about 0.001 mg to about 100
mg per kg body weight of a mammal, preferably 0.01 mg to about 50 mg
per kg, and most preferably 0.1 to 10 mg per kg, in single or divided
doses. On the other hand, it may be necessary to use dosages outside
these limits in some cases.
For use where a composition for intravenous
administration is employed, a suitable dosage range is from about 0.001
mg to about 25 mg (preferably from 0.01 mg to about 1 mg) of a compound
of Formula I per kg of body weight per day and for cytoprotective use
from about 0.1 mg to about 100 mg (preferably from about 1 mg to about
100 mg and more preferably from about 1 mg to about 10 mg) of a
compound of Formula I per kg of body weight per day.
In the case where an oral composition is employed, a
suitable dosage range is, e.g, from about 0.01 mg to about 100 mg of a
compound of Formula I per kg of body weight per day, preferably from
about 0.1 mg to about 10 mg per kg and for cytoprotective use from 0.1 mg
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to about 100 mg (preferably from about 1 mg to about 100 mg and more
preferably from about 10 mg to about 100 mg) of a compound of Formula
I per kg of body weight per day.
For the treatment of diseases of the eye, ophthalmic
preparations for ocular administration comprising 0.001-1% by weight
solutions or suspensions of the compounds of Formula I in an acceptable
ophthalmic formulation may be used.
Pharmaceutical Compositions
Another aspect of the present invention provides
pharmaceutical compositions which comprises a compound of Formula
I and a pharmaceutically acceptable carrier. The term "composition",
as in pharmaceutical composition, is intended to encompass a product
comprising the active ingredient(s), and the inert ingredients)
(pharmaceutically acceptable excipients) that make up the carrier, as
well as any product which results, directly or indirectly, from
combination, complexation or aggregation of any two or more of the
ingredients, or from dissociation of one or more of the ingredients, or
from other types of reactions or interactions of one or more of the
ingredients. Accordingly, the pharmaceutical compositions of the
present invention encompass any composition made by admixing a
compound of Formula i, additional active ingredient(s), and
pharmaceutically acceptable excipients.
Any suitable route of administration may be employed for
providing a mammal, especially a human with an effective dosage of a
compound of the present invention. For example, oral, rectal, topical,
parenteral, ocular, pulmonary, nasal, and the like may be employed.
Dosage forms include tablets, troches, dispersions, suspensions,
solutions, capsules, creams, ointments, aerosols, and the like.
The pharmaceutical compositions of the present invention
comprise a compound of Formula I as an active ingredient or a
pharmaceutically acceptable salt thereof, and may also contain a
pharmaceutically acceptable carrier and optionally other therapeutic
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ingredients. The term "pharmaceutically acceptable salts" refers to
salts prepared from pharmaceutically acceptable non-toxic bases or
acids including inorganic bases or acids and organic bases or acids.
The compositions include compositions suitable for oral,
rectal, topical, parenteral (including subcutaneous, intramuscular, and
intravenous), ocular (ophthalmic), pulmonary (aerosol inhalation), or
nasal administration, although the most suitable route in any given case
will depend on the nature and severity of the conditions being treated
and on the nature of the active ingredient. They may be conveniently
presented in unit dosage form and prepared by any of the methods well-
known in the art of pharmacy.
For administration by inhalation, the compounds of the
present invention are conveniently delivered in the form of an aerosol
spray presentation from pressurized packs or nebulisers. The
compounds may also be delivered as powders which may be formulated
and the powder composition may be inhaled with the aid of an
insufllation powder inhaler device. The preferred delivery systems for
inhalation are metered dose inhalation (MDI) aerosol, which may be
formulated as a suspension or solution of a compound of Formula I in
suitable propellants, such as fluorocarbons or hydrocarbons and dry
powder inhalation (DPI) aerosol, which may be formulated as a dry
powder of a compound of Formula I with or without additional
excipients.
Suitable topical formulations of a compound of formula I
include transdermal devices, aerosols, creams, ointments, lotions,
dusting powders, and the like.
In practical use, the compounds of Formula I can be
combined as the active ingredient in intimate admixture with a
pharmaceutical carrier according to conventional pharmaceutical
compounding techniques. The carrier may take a wide variety of forms
depending on the form of preparation desired for administration, e.g.,
oral or parenteral (including intravenous). In preparing the
compositions for oral dosage form, any of the usual pharmaceutical
_2,3_


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media may be employed, such as, for example, water, glycols, oils,
alcohols, flavoring agents, preservatives, coloring agents and the like in
the case of oral liquid preparations, such as, for example, suspensions,
elixirs and solutions; or carriers such as starches, sugars,
microcrystalline cellulose, diluents, granulating agents, lubricants,
binders, disintegrating agents and the like in the case of oral solid
preparations such as, for example, powders, capsules and tablets, with
the solid oral preparations being preferred over the liquid preparations.
Because of their ease of administration, tablets and capsules represent
the most advantageous oral dosage unit form in which case solid
pharmaceutical carriers are obviously employed. If desired, tablets may
be coated by standard aqueous or nonaqueous techniques.
In addition to the common dosage forms set out above, the
compounds of Formula I may also be administered by controlled release
means and/or delivery devices such as those described in U.S. Patent
Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; 3,630,200 and 4,008,719.
Pharmaceutical compositions of the present invention
suitable for oral administration may be presented as discrete units such
as capsules, cachets or tablets each containing a predetermined amount
of the active ingredient, as a powder or granules or as a solution or a
suspension in an aqueous liquid, a non-aqueous liquid, an oil-in-water
emulsion or a water-in-oil liquid emulsion. Such compositions may be
prepared by any of the methods of pharmacy but all methods include the
step of bringing into association the active ingredient with the carrier
which constitutes one or more necessary ingredients. In general, the
compositions are prepared by uniformly and intimately admixing the
active ingredient with liquid carriers or finely divided solid carriers or
both, and then, if necessary, shaping the product into the desired
presentation. For example, a tablet may be prepared by compression or
molding, optionally with one or more accessory ingredients.
Compressed tablets may be prepared by compressing in a suitable
machine, the active ingredient in a free-flowing form such as powder or
granules, optionally mixed with a binder, lubricant, inert diluent,


CA 02291778 1999-11-22
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surface active or dispersing agent. Molded tablets may be made by
molding in a suitable machine, a mixture of the powdered compound
moistened with an inert liquid diluent. Desirably, each tablet contains
from about 1 mg to about 500 mg of the active ingredient and each cachet
or capsule contains from about 1 to about 500 mg of the active ingredient.
The following are examples of representative
pharmaceutical dosage forms for the compounds of Formula I:
Injectable Su~~ension (I M mg/mL
l


Compound of Formula I 10


Methylcellulose 5.0


Tween 80 0,5


Benzyl alcohol 9.0


Benzalkonium chloride 1.0


Water for injection to a total volume of 1 mL
T It
/tab et


Compound of Formula I 25


Microcrystalline Cellulose 415


Povidone 14.0


Pregelatinized Starch ~,5


Magnesium Stearate 2 5
500
Capsule
Compound of Formula I 25
Lactose Powder 573.5
Magnesium Stearate
600
Aerosol°sol I'_er canist r
Compound of Formula I 24 mg
Lecithin, NF Liquid Concentrate 1.2 mg
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Trichlorofluoromethane, NF 4.025 g
Dichlorodifluoromethane, NF 12.15 g
Combination Therapy
Compounds of Formula I may be used in combination with
other drugs that are used in the treatmentJprevention/suppression or
amelioration of the diseases or conditions for which compounds of
Formula I are useful. Such other drugs may be administered, by a route
and in an amount commonly used therefor, contemporaneously or
sequentially with a compound of Formula I. When a compound of
Formula I is used contemporaneously with one or more other drugs, a
pharmaceutical composition containing such other drugs in addition to
the compound of Formula I is preferred. Accordingly, the
pharmaceutical compositions of the present invention include those that
also contain one or more other active ingredients, in addition to a
compound of Formula I. Examples of other active ingredients that may
be combined with a compound of Formula I, either administered
separately or in the same pharmaceutical compositions, include, but are
not limited to:
(a) other VLA-4 antagonists such as those described in US 5,510,332,
W097/03094, W097/02289, W096/40781, W096/22966, W096/20216,
W096/01644, W096/06108, W095/15973 and W09fi/31206; (b) steroids such
as beclomethasone, methyiprednisolone, betamethasone, prednisone,
dexamethasone, and hydrocortisone; (c) immunosuppressants such as
cyclosporin, tacrolimus, rapamycin and other FK-506 type
immunosuppressants; (d) antihistamines (H1-histamine antagonists)
such as bromopheniramine, chlorpheniramine, dexchlorpheniramine,
triprolidine, clemastine, diphenhydramine, diphenylpyraline,
tripelennamine, hydroxyzine, methdilazine, promethazine,
trimeprazine, azatadine, cyproheptadine, antazoline, pheniramine
pyrilamine, astemizole, terfenadine, loratadine, cetirizine,
fexofenadine, descarboethoxyloratadine, and the like; (e) non-steroidal
anti-asthmatics such as ~i2-agonists (terbutaline, metaproterenol,
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fenoterol, isoetharine, albuterol, bitolterol, salmeterol and pirbuterol),
theophylline, cromolyn sodium, atropine, ipratropium bromide,
leukotriene antagonists (zafirlukast, montelukast, pranlukast,
iralukast, pobilukast, SKB-106,203), leukotriene biosynthesis inhibitors
(zileuton, BAY-1005); (f) non-steroidal antiinflammatory agents
(NSAIDs) such as propionic acid derivatives (alminoprofen,
benoxaprofen, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen,
flurbiprofen, ibuprofen, indoprofen, ketoprofen, miroprofen, naproxen,
oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid, and
tioxaprofen), acetic acid derivatives (indomethacin, acemetacin,
alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac,
furofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin,
zidometacin, and zomepirac), fenamic acid derivatives (flufenamic acid,
meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid),
biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicams
(isoxicam, piroxicam, sudoxicam and tenoxican), salicylates (acetyl
salicylic acid, sulfasalazine) and the pyrazolones (apazone,
bezpiperylon, feprazone, mofebutazone, oxyphenbutazone,
phenylbutazone); (g) cyclooxygenase-2 (COX-2) inhibitors such as
celecoxib; (h) inhibitors of phosphodiesterase type IV (PDE-IV); (i)
antagonists of the chemokine receptors, especially CCR-1, CCR-2, and
CCR-3; (j) cholesterol lowering agents such as HMG-CoA reductase
inhibitors (Iovastatin, simvastatin and pravastatin, fluvastatin,
atorvastatin, and other statins), sequestrants (cholestyramine and
colestipol), nicotinic acid, fenofibric acid derivatives (gemfibrozil,
clofibrat, fenofibrate and benzafibrate), and probucol; (k) anti-diabetic
agents such as insulin, sulfonylureas, biguanides (metformin), a-
glucosidase inhibitors (acarbose) and glitazones (troglitazone,
pioglitazone, englitazone, MCC-555, BRL49653 and the like); (1)
preparations of interferon beta (interferon beta-la, interferon beta-lb);
(m) anticholinergic agents such as muscarinic antagonists
(ipratropium bromide); (n) other compounds such as 5-aminosalicylic
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acid and prodrugs thereof, antimetabolites such as azathioprine and 6-
mercaptopurine, and cytotoxic cancer chemotherapeutic agents.
The weight ratio of the compound of the Formula I to the
second active ingredient may be varied and will depend upon the
effective dose of each ingredient. Generally, an effective dose of each will
be used. Thus, for example, when a compound of the Formula I is
combined with an NSAID the weight ratio of the compound of the
Formula I to the NSAID will generally range from about 1000:1 to about
1:1000, preferably about 200:1 to about 1:200. Combinations of a
compound of the Formula I and other active ingredients will generally
also be within the aforementioned range, but in each case, an effective
dose of each active ingredient should be used.
Compounds of the present invention may be prepared by
procedures illustrated in the accompanying schemes. In the first
method (Scheme 1), a resin-based synthetic strategy is outlined where
the resin employed is represented by the ball ( ~ ). An N-Fmoc-
protected amino acid derivative A (Fmoc = fluorenylmethoxycarbonyl)
is loaded on to the appropriate hydroxyl-containing resin using
dicyclohexylcarbodiimide (DCC) and 1-hydroxybenzotriazole (HOBt)
in dimethylformamide (DMF) to give B. The Fmoc protecting group
is removed with piperidine in DMF to yield free amine C. The next
Fmoc-protected amino acid derivative D is coupled to C employing
standard peptide (in this instance, 2-(1H-benzotriazol-1-yl)-1,1,3,3-
tetramethyluronium hexafluorophosphate (HBTU), HOBt, and N,N-
diisopropylethylamine (DIEA) in DMF) to yield dipeptide E. The
Fmoc group is removed with piperidine in DMF to yield the free
amine F. An acid chloride or isocyanate derivative is reacted with F
in the presence of DIEA to yield G. The final product is removed from
the resin with strong acid (in this instance, trifluoroacetic acid (TFA)
in the presence of thioanisole and dithiane) to yield compounds of the
present invention H.
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Scheme 1
Na O HO Ns O
Fmoc' '
~OH Fmoc p--.~,~.
Ra RS DCC, HOBt
DMF Ra R5 B
_A
Rs O HBTu, HOBt,
HN N DIEA, DMF
H~ ~O
DMF Ra Rs R~
R~ ~~.RB
C A~ /~'~ OH
R~ N RL II
Rs B-y Z Ra R3 O Fmoc O D
N
ANN~ ~O--~- piperidine
R' II
Fmoc O Ra R5 E DMF
R'
_ R~-Y-CI
R ~ ~ j Ra R3 O or
w N ~~ O--~,.,~- ::
A N R - N=C=O
R~
H O Ra R5 DIEA
F
R~
R ~ ~ j R8 Ra O TFA, PhSCH3,
N~O~ , HSCH2CH2SH
NR
Y O Ra Rs
R' ~ R~
Rs B-~ j Rs R3 O
N
~OH
R~
z II Ra R5
RiY O H_
-29-


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WO 98/53814 PCT/US98/10940
In the second method (Scheme 2}, standard solution
phase synthetic methodology is outlined. An N-Boc-protected amino
acid derivative A (Boc = tert-butyloxycarbonyl) is treated with tert-
butyl 2,2,2-trichloroacetimidate in the presence of boron trifluoride
etherate to yield tert-butyl ester followed by treatment with strong acid
(HCl in ethyl acetate or sulfuric acid in t-butyl acetate) to yield the free
amine B which is subsequently coupled to Cbz-protected amino acid
derivative C (Cbz = carbobenzyloxy) in the presence of 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC),
HOBt, and N-methylmorpholine (NMM) in methylene chloride
(Methylene chloride) to yield dipeptide D. Catalytic hydrogenation of
D in the presence of a palladium-on-carbon (Pd/C) catalyst yields E.
Reaction of E with an acylchloride or isocyanate in the presence of
DIEA and 4-dimethylaminopyridine (DMAP) yields F which is
subsequently reacted with strong acid (TFA) to yield the desired
product G.
-30-


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Scheme 2
NH
Rs O ~~~\ ~~~ R3 O
,N ~ CI3C O N
Boc OH H~ ~O
R4 R5 Bycl hte~~ eH2Cl2 R4 R5
A _B
R'
_ 8
Rs~ I. Z/ R R~
A~N~OH Rs g_LZ R8 R3 O
C i 'R j~ \ %
Cbz O A ~ N ~ H2, Pd/C
N
R
EDC, HOBt Cbz O R4 R5 CH30H
NMM, CH2CI2 D
R~ R1-Y-Cf
Rs ByZ Rs R3 O or
N ~ R~-N=C=O
Aw ~O
R~ DIEA
H O R4 R5 4-DMAP
E
R' R'
Rs B-yZ RB R3 O Rs ByZ Ra R3 O
p ~ N k q ~ N
~O OH
N N
Ri~Y R p R4 Rs TFA, CH2CI2 R1~Y R p R4 Rs
F Sa
-31-


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WO 98/53814 PCT/US98/10940
GENERAL PROCEDURE FOR THE SOLID-PHASE SYNTHESIS_O_F
COMPOUNDS OF FORMULA 1
Step A. Loading of N-Fmoc-amino acid derivatives onto resins
N-Fmoc-amino acids were loaded on either Wang~
(Calbiochem-Novabiochem Corp.) or Chloro (2-chlorotrityl) resin.
Wang~ resin, typically 0.3 mmol, was washed with
dimethylformamide three times. A solution of N-Fmoc-amino acid
(0.3 mmol) in dimethylformamide (3 mL) was transferred to the pre-
swollen Wang~ resin. Dicyclohexylcarbodiimide (0.3 mmol) and 1-N-
hydroxybenztriazole (0.3 mmol) was added and the mixture gently
swirled for 2 hours. Following filtration, the resin was sequentially
washed with dimethylformamide (3 times) and dichloromethane (3
times). The amino acid substitution value obtained after vacuum
drying typically ranged between 0.07 to 0.1 mmol.
Alternatively, Chloro (2-chorotrityl) resin, typically 0.2
mmol, was pre-swollen in dimethylformamide. A solution of N-
Fmoc-amino acid {0.2 mmol) in dimethylformamide (3 ml) was added
to the resin, followed by the addition of N,N-diisopropylethylamine(0.4
mmol). The resin was gently stirred for 2 hours, filtered and washed
sequentially with dimethylformamide (3 times) and dichloromethane
(3 times). The resin was finally washed with 10% methanol in
dichloromethane and vacuum dried. The amino acid substitution
value obtained after vacuum drying typically ranged between 0.05 to
0.1 mmol.
Step B. Deprotection of the N-Fmoc ~rouu.
The N-Fmoc protecting group was removed from the
resin from Step A by treatment with 20% piperidine in
dimethylformamide for 30 minutes. Following filtration, the resin
was washed sequentially with dimethylformamide (3 times),
dichloromethane (1 time) and dimethylformamide (2 times) and used
in the subsequent reaction.
-32-


CA 02291778 1999-11-22
WO 98/53814 PCT/CTS98/10940
Step C. Coupling of thp next N Fm~r. amino acid d~Privative
A solution of the next desired N-Fmoc-amino acid
derivative (0.4 mmol) in dimethylformamide (2 mL) was mixed with
2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate (0.4 mmol), 1-hydroxybenzotriazole (0.4 mmol)
and diisopropylethylamine (0.6 mmol). This solution was transferred
to resin from Step B and typically allowed to react for 2 hours.
Couplings were monitored by ninhydrin reaction. The coupling
mixture was filtered and the resin washed with dimethylformamide
(3 times) and used in the subsequent reaction.
Step D. Denrotection of the N-Fmoc ~rouu.
The N-Fmoc protecting group was removed from the
resin from Step C by the procedure described in Step B and used in
the subsequent reaction.
Step E. Acvlation (or sulfonyl)at»n) of the terminal ino~rou
The desired N-terminal capping reagent (sulfonyl)
chloride or acyl chloride, or isocyanate) (0.4 mol) was dissolved in
dimethylformamide (2 ml), mixed with N,~T-
diisopropylethylamine(0.8 mmol) and added to the resin from Step D.
After approximately two hours, the resin was sequentially washed
with dimethylformamide (3 times) and dichloromethane (3 times).
Step F. Cleavaee of the desired products from the reR~n~
The final desired products were cleaved from the resins
from Step E by gently stirring with a solution of trifluoroacetic
acidahioanisole:ethanedithiol (95:2.5:2.5); 3 hours far Wang~ resin
and 30 minutes for the Chloro (2-chorotrityl) resin. Following
filtration, the solvents were removed by evaporation and the residue
dissolved in acetonitrile (3 mL). Insoluble material was removed by
filtration. The final products were purified by reverse phase
-33-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
chromatography with a linear gradient of buffer A (0.1%
trifluoroacetic acid in water) and buffer B (0.1% trifluoroacetic acid in
acetonitrile) and isolated by lyophilization. Molecular ions were
obtained by electrospray ionization mass spectrometry or matrix-
assisted laser desorption ionization time-of flight mass spectrometry
to confirm the structure of each peptide.
The following compounds were prepared by the general
procedures described above using the appropriate amino acid derivatives
and acyl or sulfonyl) chloride or alkyl or aryl isocyanate. These
examples are provided to illustrate the present invention and are not to
be construed as limiting its scope in any manner.
Ex. Compound Name MS
(1) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 491
tetrahydroisoquinoline-3(S)-carbonyl-(L)-
leucine
(2) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-534


tetrahydroisoquinoline-3(S)-carbonyl-(L)-


arginine


(3) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-507


tetrahydroisoquinoline-3(S)-carbonyl-(L)-


glutamic acid


(4) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-435


tetrahydroisoquinoline-3(S)-carbonyl-glycine


(5) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-575


tetrahydroisoquinoline-3(S)-carbonyl-(L)-(
1-


naphthyl)alanine


(6) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-491


tetrahydroisoquinoline-3(S)-carbonyl-(L)-a-t-


butylglycine


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
(7) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 531
tetrahydroisoquinoline-3(S)-carbonyl-(L)-3-(2-
thienyl)alanine
(8) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 531
tetrahydroisoquinoline-3(S)-carbonyl-(L)-
cyclohexylalanine
(9) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 575
tetrahydroisoquinoline-3(S)-carbonyl-(L)-3-(2-
naphthyl)alanine
(10) N-(3,3-diphenylpropanoyl)-1,2,3,4-tetrahydro 498
isoquinoline-3(S)-carbonyl-(L)-norleucine
(11) N-(2,4-dinitrobenzenesulfonyl)-1,2,3,4- 521
tetrahydroisoquinoline-3(S)-carbonyl-(L)-
norleucine
(12)N-(3,4-dimethoxybenzenesulfonyl)-1,2 601
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-3,3-


diphenylalanine


(13)N-(3,4-dimethoxybenzenesulfonyl)-1,2,3 537
4-


,
tetrahydroisoquinoline-3(S)-carbonyl-1
2
3
4-


,
,
,
tetrahydroisoquinoline-3-carboxylic acid


(14)N-(3,4-dimethoxybenzenesulfonyl)-1,2 475
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-proline


(15)N-dansyl-1,2,3,4-tetrahydroisoquinoline-3(S)-511


carbonyl-(L)-norleucine


(16)N-(2-naphthalenesulfonyl)-1,2,3,4-tetrahydro481


isoquinoline-3(S)-carbonyl-(L)-norleucine


(17)N-(4-methoxybenzenesulfonyl)-1,2,3,4-tetrahydro461


isoquinoline-3(S)-carbonyl-(L)-norleucine


(18)N-(4-phenylbenzoyl)-1,2,3,4-tetrahydro 471


isoquinoline-3(S)-carbonyl-(L)-norleucine


(19)N-(3,4-dimethylbenzenesulfonyl)-1,2 481
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-cysteine


-35-


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(20) N-(4-t-butylbenzenesulfonyl)-1,2,3,4-tetrahydro 487
isoquinoline-3(S)-carbonyl-(L)-norleucine
(21) N-(2,5-dichlorobenzenesulfonyl)-1,2,3,4- 498
tetrahydro isoquinoline-3(S)-carbonyl-(L)-
norleucine
(22) N-(2-mesitylenesulfonyl)-1,2,3,4-tetrahydro473


isoquinoline-3(S)-carbonyl-(L)-norleucine


(23) N-(p-toluenesulfonyl)-1,2,3,4-tetrahydro444


isoquinoline-3(S)-carbonyl-{L)-norleucine


(24) N-{4-chlorobenzenesulfonyl)-1,2,3,4-tetrahydro465


isoquinoline-3(S)-carbonyl-(L)-norleucine


(25) N-(N'-acetylsulfanilyl)-1,2,3,4-tetrahydro488


isoquinoline-3(S)-carbonyl-(L)-norleucine


(26) N-(4-fluorobenzenesulfonyl)-1,2,3,4-tetrahydro449


isoquinoline-3(S)-carbonyl-(L)-norleucine


(27) N-(1-naphthalenesulfonyl)-1,2,3,4-tetrahydro481


isoquinoline-3(S)-carbonyl-(L)-norleucine


(28) N-(benzylsulfonyl)-1,2,3,4- 445


tetrahydroisoquinoline-3(S)-carbonyl-(L)-


norleucine
(29) N-(4-nitrobenzenesulfonyl)-1,2,3,4-tetrahydro 476
isoquinoline-3(S)-carbonyl-(L)-norleucine
(30) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 525
tetrahydroisoquinoline-3(S)-carbonyl-(L)-
phenylalanine
(31) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 506
tetrahydroisoquinoline-3(S)-carbonyl-(L)-
glutamine
(32) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 570
tetrahydroisoquinoline-3(S)-carbonyl-(L)-(4-
nitrophenyl)alanine
-36-


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WO 98/53814 PCT/IJS98/10940
(33) N-(3,4-dimethoxybenzenesulfonyl)-1 492
2
3
4-


,
,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-


asparagine


(34) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-509


tetrahydroisoquinoline-3(S)-carbonyl-(L)-


methionine


(35) N-(3,4-dimethoxybenzenesulfonyl)-1,2 539
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-


homophenylalanine


(36) N-(3,4-dimethoxybenzenesulfonyl)-1,2 491
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(D)-


norleucine


(37) N-(3,4-dimethoxybenzenesulfonyl)-1,2 543
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-(4-


fluorophenyl)alanine


(38) N-(3-toluenesulfonyl)-1,2,3,4-tetrahydro4~5


isoquinoline-3(S)-carbonyl-(L)-norleucine


(39) N-(4-trifluoromethylbenzenesulfonyl)-1,2,3499
4-


,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-


norleucine


(40) N-(4-n-propylbenzenesulfonyl)-1,2,3,4-tetrahydro473


isoquinoiine-3(S)-carbonyl-(L)-norleucine


(41) N-(4-isopropylbenzenesulfonyl)-1,2,3 473
4-


,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-


norleucine


(42) N-(2,6-dichlorobenzenesulfonyl)-1 4gg
2
3
4-


,
,
,
tetrahydro isoquinoline-3(S)-carbonyl-(L)-


norleucine
(43) N-(4-ethylbenzenesulfonyl)-1,2,3,4-tetrahydro 459
isoquinoline-3(S)-carbonyl-(L)-norleucine
(44) N-(2,4-difluorobenzenesulfonyl)-1,2,3,4- q$7
tetrahydro isoquinoline-3(S)-carbonyl-(L)-
norleucine
-37-


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WO 98/53814 PCT/US98/10940
(45) N-(2-cyanobenzenesulfonyl)-1,2,3,4-tetrahydro456


isoquinoline-3(S)-carbonyl-(L)-norleucine


(46) N-(4-tert-amylbenzenesulfonyl)-1,2,3,4- 501


tetrahydro isoquinoline-3(S)-carbonyl-(L)-


norleucine


(47) N-(4-chloro-3-nitrobenzenesulfonyl)-1,2,3,4-510


tetrahydroisoquinoline-3(S)-carbonyl-{L)-


norleucine


(48) N-(3-cyanobenzoyl)-1,2,3,4-tetrahydro 420


isoquinoline-3(S)-carbonyl-(L)-norleucine


(49) N-(3,5-dichlorobenzenesulfonyl)-1 499
2
3
4-


,
,
,


tetrahydroisoquinoline-3(S)-carbonyl-(L)-


norleucine


(50) N-(3,4-dichlorobenzenesulfonyl)-1,2,3,4-499


tetrahydroisoquinoline-3(S)-carbony(L)-


norleucine


(51) N-(2-trifluoromethylbenzenesulfonyl)-1,2,3,4-499


tetrahydroisoquinoline-3(S)-carbonyl-(L)-


norleucine


(52) N-(2,3-dichlorobenzenesulfonyl)-1,2 499
3
4-


,
,
tetrahydro isoquinoline-3(S)-carbonyl-(L)-


norleucine
(53) N-(2,4-dichlorobenzenesulfonyl)-1,2,3,4- 499
tetrahydro isoquinoline-3(S)-carbonyl-(L)-
norleucine
(54) N-(2,5-dimethoxybenzenesulfonyl)-1,2,3,4- 49I
tetrahydroisoquinoline-3(S)-carbonyl-(L)-
norleucine
(55) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 465
tetrahydroisoquinoline-3(S)-carbonyl-(L)-serine
(56) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4- 491
tetrahydroisoquinoline-3(S )-carbonyl-(L)-
isoleucine
-38-


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WO 98/53814 PCT/US98/10940
(57) N-(3,4-dimethoxybenzenesulfonyl)-1,2 56
3
4-


,
,
tetrahydroisoquinoline-3{S)-carbonyl-(L)-


tryptophan


(58) N-(2,1,3-benzothiadiazole-4-sulfonyl)-1,248.9
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-


tryptophan


(59) N-(3,4-dimethoxybenzenesulfonyl)-1,2 5~
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-3-(3-


pyridyl)alanine


(60) N-(3,4-dimethoxybenzenesulfonyl)-1,2 603
3
4-


,
,
tetrahydroisoquinoline-3(S)-carbonyl-{L)-3-(2-


naphthyl)alanine, ethyl ester


(61) N-acetyl-1,2,3,4-tetrahydroisoquinoline-3(S)-333


carbonyl-(L)-norleucine


(62) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3 491
4-


,
tetrahydroisoquinoline-3(R)-carbonyl-(D)-


norleucine


{63) N-propionyl-(L)-prolyl-1,2,3,4-tetrahydro348


isoquinoline-3(S)-carbonyl-(L)-norleucine


(64) N-(4-cyanobenzenesulfonyl)-1,2,3,4-tetrahydro456


isoquinoline-3(S)-carbonyl-(L)-norleucine


(65) N-(benzenesulfonyl)-1,2,3,4-tetrahydro qg1


isoquinoline-3(S)-carbonyl-(L)-norleucine


(66) N-(3-nitrobenzenesulfonyl)-1,2,3,4-tetrahydro476


isoquinoline-3(S)-carbonyl-(L)-norleucine


(67) N-(3-trifluoromethylbenzenesulfonyl)-1,2499
3
4-


,
,
tetrahydroisoquinoline-3(S )-carbonyl-(L)-


norleucine


(68) N-(2-thienylsulfonyl)-1,2,3,4-tetrahydro437


isoquinoline-3(S)-carbonyl-(L)-norleucine


(69) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3 505
4-


,
tetrahydroisoquinoline-3(S)-carbonyl-(L)-N-


methylleucine


-39-


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WO 98/53814 PCTlUS98/10940
(70) N-(3,4-dimethoxybenzenesulfonyl)-1,2,3,4-535


tetrahydraisoquinoline-3(S)-carbonyl-(L)-


citrulline


(71) N-(4-iodobenzenesulfonyl)-1,2,3,4-tetrahydro557


isoquinoline-3(S)-carbonyl-(L)-norleucine


(72) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-(3-613


iodo)tyrosine


(73) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-472


(3-pyridyl)alanine


(74) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-471


phenylalanine


(75) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-453


glutamic acid


(76) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-480


arginine


(77) N-(N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl)-1-549


amino-cyclopentane-1-carboxylic acid


(78) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-541


(3,4-dichlorophenyl)alanine


(79) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-549


(2-naphthyl)alanine, ethyl ester


(80) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-550


(4-bromophenyl)alanine


(81) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-516


(4-nitrophenyl)alanine


(82) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-478


(4-thiazolyl)alanine


(83) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-507


{2-chlorophenyl)alanine


(84) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-507


(4-chlorophenyl)alanine


(85) N-(3,5-dichlorobenzenesulfonyl)-{L)-prolyl-(L)-3-496


(4-cyanophenyl)alanine


_40-


CA 02291778 1999-11-22
WO 98/53$14 PCT/US98/10940
(86) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-586


tyrosine, O-sulfate


(87) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-739


3,5-diiodotyrosine


(88) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-488


tyrosine


(89) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-438


aspartic acid


(90) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-510


tryptophan


(9I) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-454


methionine


(92) N-(3,4-dimethoxybenzenesulfonyl)-(L)-prolyl-(L)-429


norleucine


(93) N-(3,5-di(trifluoromethyl)benzenesulfonyl)-(L)-589


prolyl-(L)-3-(2-naphthyl)alanine


(94) N-(3,4-dimethoxybenzenesulfonyl)-(L)-thiaprolyl-531


(L)-3-(2-naphthyl)alanine


(95) N-(3,4-dimethoxybenzenesulfonyl)-(L)-thiaprolyl-447


(L)-norleucine


(96) N-[4-(N'-2-toluylureido)phenylacetyl]-(L)-5g7


thiaprolyl-(L)-3-(2-naphthyl)alanine


(97) N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-539


(L)-3-(2-naphthyl)alanine


(98) N-(3,4-dimethoxybenzenesulfonyl)-(L)-pipecolyl-443


(L)-norleucine


(99) N-(3,4-dimethoxybenzenesulfonyl)-(L)-pipecolyl-471


(L)-norleucine, ethyl ester


(100)N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-499


homophenylaianine


(101)N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-626


( 3-iodo)tyrosine


-41-


CA 02291778 1999-11-22
WO 98/53814 PCT/US98/10940
(102) N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)- 535
3-(2-naphthyl)alanine


(103)N-[4-(N'-2-toluylureido)phenylacetyl]-(L)-593


pipecoliny(L)-3-(2-naphthyl)alanine


(104)N-(3,5-di(trifluoromethyl)benzenesulfonyl)]-(L)-603


pipecolyl-(L)-3-(2-naphthyl)alanine


(105)N-(3,4-dimethoxybenzenesulfonyl)-(L)-pipecolyl-555


(L)-3-(2-naphthyl)alanine, ethyl ester


(106)N-(3,4-dimethoxybenzenesulfonyl)-(L)- 483


octahydroisoquinoline-3-carbonyl-(L)-norleucine


( N-(3,4-dimethoxybenzenesulfonyl)-azetidine-2-4I5
107)


carbonyl-(L)-norleucine


(108)N-(3,5-dichlorobenzenesulfonyl)-(L)-4(S)-537


hydroxyprolyl-(L)-3-(2-naphthyl)alanine


(109)N-(3,4-dimethoxybenzenesulfonyl)-(L)-4(S)-445


hydroxyprolyl-(L)-norleucine


( N-(3,4-dimethoxybenzenesulfonyl)-(L)-3,4-427
110)


dehydroprolyl-(L)-norleucine


(111)N-(3-bis(N,N-


benzenesulfonyl)aminobenzenesulfonyl)-(L)-


prolyl-(L)-norleucine


( N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-3-472.2
112)


(4-pyridyl)alanine


(113)N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-536.1


aminoprolyl-(L)-3-(2-naphthyl)alanine


(114)N-(3,5-dichlorobenzenesulfonyl)-(L)-3,4- 487.2


dehydroprolyl-(L)-4-fluorophenylalanine


(115)N-(3-chlorobenzenesuifonyl)-(L)-prolyl-(L)-4-455.1


fluorophenylalanine


(116)N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-505.2


hydroxyprolyl-(L)-4-fluorophenylalanine


(117)N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-505.0


(L)-tyrosine
-42-


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WO 98/53814 PCT/US98/10940
(118) N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl- 631.0
(L)-3-iodotyrosine


(119)N-(3-fluorobenzenesulfonyl)-(L)-thiaprolyl-(L)-3-489.3


(2-naphthyl)alanine


(120)N-(3-fluorobenzenesulfonyl)-(L)-pipecolyl-(L)-3-(2-485.4


naphthyl)alanine


(121)N-(3-fluorobenzenesulfonyl)-(L)-thiaprolyl-(L)-4-457.2


fluorophenylalanine


(122)N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-4-439.2


fluorophenylalanine


(123)N-(3-chlorobenzenesulfonyl)-(L)-3,4- 453.3


dehydroprolyl-(L)-4-fluorophenylalanine


(124)N-(3-fluorobenzenesulfonyl)-(L)-4(R)- 4,55,0


hydroxyprolyl-(L)-4-fluorophenylalanine


(125)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 471.0


hydroxyprolyl-(L)-4-fluorophenylalanine


(126)N-(3,5-dichlorobenzenesulfonyl)-(L)-pipecolyl-(L)-503.1


4-fluorophenylalanine


(127)N-(3-fluorobenzenesulfonyl)-(L)-3,4- 435.3


dehydroprolyl-(L)-tyrosine


(128)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-prolyl-493.2


(L)-tyrosine


(129)N-(3-fluorobenzenesulfonyl)-(L)-4(R)- 453.2


hydroxyprolyl-(L)-tyrosine


(130)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 469.2


hydroxyprolyl-(L)-tyrosine


(131)N-(3-fluorobenzenesulfonyl)-(L)-pipecolyl-(L)-4-453.3


fluorophenylalanine


(132)N-(3-fluorobenzenesulfonyl)-(L)-4(R)- 509.1


hydroxyprolyl-(L)-tyrosine, O-tert-butyl
ether


(133)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 525.3


hydroxyprolyl-(L)-tyrosine, O-tert-butyl
ether


_43_


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(134) N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4- 491.1
dehydroprolyl-(L)-tyrosine


(135)N-(3,5-dichlorobenzenesulfonyl)-(L)-3(S)-methyl-503.1


prolyl-(L)-4-fluorophenylalanine


(136)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-485.1


dehydroprolyl-(L)-tyrosine


(13?)N-(3-fluorobenzenesulfonyl)-(L)-3,4- 491.1


dehydroprolyl-(L)-tyrosine, O-tert-butyl
ether


(I38)N-(3-chlorobenzenesulfonyl)-(L)-3,4- 507.3


dehydroprolyl-(L)-tyrosine, O-tert-butyl
ether


(139)N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-469.1


prolyl-(L)-4-fluorophenylalanine


(140)N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-467.3


prolyl-(L)-tyrosine


(141)N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl-523.2


prolyl-(L)-tyrosine, O-tert-butyl ether


(142)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-50L0


prolyl-(L)-tyrosine


(143)N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-3-563.1


iodotyrosine


(144)N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-3-579.0


iodotyrosine


(145)N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-3-421.1


phenylalanine


(146)N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-437.3


phenylalanine


(147)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-471.2


phenylalanine


(148)N-(3-fluorobenzenesulfonyl)-(L)-4(R)- 437.3


hydroxyprolyl-(L)-phenylalanine


(149)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 453.2


hydroxyprolyl-(L)-phenylalanine


_4,~_


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PCT/US98/10940
(150)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-476.1


dehydroprolyl-(L)-3-(4-pyridyl)alanine


(151)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-495.9


thiaprolyl-(L)-3-(4-pyridyl)alanine


(152)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-3,4-492.9


dehydroprolyl-(L)-4-fluorophenylalanine


(153)N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-487.1


hydroxyprolyl-(L)-phenylalanine


(154)N-(3-trifluoromethylbenzenesulfonyl)-(L)-prolyl-489.3


(L)-4-fluorophenylalanine


(155)N-(3-trifluoromethylbenzenesulfonyl)-(L)-507.0


thiaprolyl-(L)-4-fluorophenylalanine


(156)N-(3-fluorobenzenesulfonyl)-(L)-3,4- 437.1


dehydroprolyl-(L)-4-fluorophenylalanine


(157)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-567.0


tyrosine, O-phosphoric acid


(158)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- q.6g,3


aminoprolyl-(L)-tyrosine


(159)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-510.9


thiaprolyl-(L)-tyrosine


(160)N-(Nl-methyl-4-imidazolesulfonyl)-(L)-prolyl-(L)-425.3


4-fluorophenylalanine


(161)N-(3,5-dichlorobenzenesulfonyl)-(D)-prolyl-(D)-4-489.1


fluorophenylalanine


(162)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-4(R)-492.9


aminoprolyl-(L)-3-(4-pyridyl)alanine


(163)N-(5-(5-trifluoromethyl-2-pyridylsulfonyl)-2-636.1


thiophenesulfonyl)-(L)-prolyl-(L)-4-


fluorophenylalanine


(164)N-(5-(N-(4-chlorobenzoyl)aminomethyl))-2-575.1


thiophenesulfonyl)-(L)-prolyl-(L)-4-


fluorophenylalanine


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(165) N-(5-(3-(1-methyl-5-trifluoromethyl-pyrazoyl))-2- 594.0
thiophenesulfonyl)-(L)-prolyl-(L)-4-
fluorophenylaianine
(166) N-(3-fluorobenzenesulfonyl)-2(S)-methylprolyl- 507.3
(L)-O-tert-butyl-tyrosine
(167)N-(3-fluorobenzenesulfonyl)-(L)-4(R)- 454.2


aminoprolyl-(L)-4-fluorophenylalanine


(168)N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-504.3


aminoprolyl-(L)-4-fluorophenylalanine


(169)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 470.1


aminoprolyl-(L)-4-fluorophenylalanine


(170}N-(3,5-dichlorobenzenesulfonyl)-(L)-4(S}-504.0


aminoprolyl-(L)-4-fluorophenylalanine


(171)N-(3-chlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-4-473.3


fluorophenylalanine
(172) N-(4-bromo-5-chloro-2-thiophenesulfonyl)-(L)- 540.9
prolyl-(L)-4-fluorophenylalanine


(173)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-prolyl-513.0


(L)-4-fluorophenylalanine


(174)N-(3,5-dichlorobenzenesulfonyl)-(L)-thiaprolyl-756.7


(L)-3,5-diiodotyrosine


(175)N-(5-benzoylaminomethyl-2-thiophenesulfonyl)-560.1


(L)-prolyl-(L)-4-fluorophenylalanine


(176)N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-O-tent-509.3


butyl-tyrosine


(177)N-(5-benzenesulfonyl-2-thiophenesulfonyl)-(L)-567.0


prolyl-(L)-4-fluorophenylalanine


(178)N-(3-bromo-5-chloro-2-thiophenesulfonyl)-(L)-540.9


prolyl-(L)-4-fluorophenylalanine


(179)N-(3-chlorobenzenesulfonyl)-(L)-3,4- 451.2


dehydroprolyl-(L)-tyrosine


(180)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-485.3


homophenylalanine


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(I81)N-(4-benzenesulfonyl-2-thiophenesulfonyl)-(L)-621.1


prolyl-(L)-O-tert-butyl-tyrosine


(182)N-(5-benzoylaminomethyl-2-thiophenesulfonyl)-614.2


{L)-prolyl-(L)-O-tert-butyl-tyrosine


(183)N-(traps-2-phenyl-ethylene-sulfonyl)-(L)-prolyl-501.3


(L)-O-tent-butyl-tyrosine


{184)N-(5-benzenesulfonyl-2-thiophenesulfonyl)-(L)-621.1


prolyl-{L)-O-tent-butyl-tyrosine


{185)N-(3-fluorobenzenesulfonyl)-(L)-thiaprolyl-(L)-O-511.2


tent-butyl-tyrosine


( N-(benzylsulfonyl)-(L)-prolyl-(L)-O-tert-butyl-489.3
186)


tyrosine


(187)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-426.2


cysteine, amide


(188)N-(1-methyl-4-imidazolylsulfonyl)-(L)-prolyl-(L)-479.1


O-tert-butyl-tyrosine


(189)N-(4-(N-(4-dimethylaminophenyl)diazo)- 622.0


benzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-


tyrosine


(190)N-(5-(4-trifluoromethylbenzenesulfonyl)-2-690.2


thiophenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-


tyrosine


(191)N-(3-bromobenzenesulfonyl)-(L)-prolyl-(L)-O-tent-553.2


butyl-tyrosine


(192)N-(4-methylsulfonyl-benzenesulfonyl)-(L)-prolyl-499.2


(L)-4-fluorophenylalanine


(193)N-{4-methoxybenzenesulfonyl)-(L)-prolyl-(L)-O-505.2


tert-butyl-tyrosine


(194)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-prolyl-495.0


(L)-3-fluorophenylalanine


(195)N-(5-chloro-2-thiophenesulfonyl)-(L)-prolyl-(L)-4-461.1


fluorophenylalanine


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(196) N-(3-chlorobenzenesulfonyl)-(L)-thiaprolyl-(L)- 471.0
tyrosine


(197)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-558.6


methylprolyl-(L)-O-tert-butyl-tyrosine


(198)N-(1(R)-(+)-10-camphorsulfonyl)-(L)-prolyl-(L)-O-549.3


tert-butyl-tyrosine


(199)N-(1(S)-(+)-10-camphorsulfonyl)-(L)-prolyl-(L)-O-549.3


tert-butyl-tyrosine


(200)N-(3,4-methylenedioxy-phenylacetyl)-(L)-proiyl-497.2


(L)-O-tert-butyl-tyrosine


(201)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 551.0


hydroxyprolyl-(L)-tyrosine-O-sulfate


(202)N-(3-chlorobenzenesulfonyl)-(L)-thiaprolyl-(L)-553.7


tyrosine-O-sulfate


(203)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-427.2


cysteine


(204)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-N-451.2


methyl-isoleucine


(205)N-(3,5-dichlorobenzenesulfonyl)-(L)-4(R)-558.3


aminoprolyl-(L)-O-tert-butyl-tyrosine


(206)N-(3-chlorobenzenesulfonyl)-(L)-4(R)- 524.4


aminoprolyl-(L)-O-tert-butyl-tyrosine


(207)N-(3-cyanobenzenesulfonyl)-(L)-prolyl-(L)-444.3


tyrosine


(208)N-benzenesulfonyl-(L)-prolyl-(L)-O-tert-butyl-475.5


tyrosine


(209)N-(4-methylsulfonylbenzenesulfonyl)-(L)-prolyl-553.2


(L)-O-tent-butyl-tyrosine


(210)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-4(R)-564.3


aminoprolyl-(L)-O-tert-butyl-tyrosine


(211)N-(4,5-dichloro-2-thiophenesulfonyl)-(L)-4(R)-510.1


aminoprolyl-(L)-4-fluorophenylalanine


_4g-


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(212)N-(9-fluorenylmethyloxycarbonyl)-(L)-prolyl-(L)-485


phenylalanine


(213)N-(benzenesulfonyl)-(L)-prolyl-(L)-phenylalanine403


(214)N-(n-octyl-1-sulfonyl)-(L)-prolyl-(L)- 418


phenylalanine


(215)N-(3-fluorobenzenesulfonyl)-(L)-5(R)-phenyl-515


prolyl-(L)-4-fluorophenylalanine


(216)N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-phenyl-582


prolyl-(L)-4-iodophenylalanine


(217)N-(3,5-dichlorobenzenesulfonyl)-1,2,3 ~g
4-


,
tetrahydro isoquinoline-1-carbonyl-(L)-4-


fluorophenylalanine


(218)N-(3,5-dichlorobenzenesulfonyl)-1,3-dihydro554


isoindolyl-1-carbonyl-(L)-4-fluorophenylalanine


(219)N-(4-(fluorescien-4-carbonylamino)benzene879.2


sulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine


(220)N-(3-ethoxycarbonyl-benzenesulfonyl)-(L)-prolyl-547.2


(L)-O-tent-butyl-tyrosine


(221)N-(4-iodobenzenesulfonyl)-(L)-prolyl-(L)-4-633.0


benzoyl-phenylalanine


(222)N-(3-(4-benzophenonyl-carbonylamino)- 6gg,2


benzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-


tyrosine


(223)N-(3-(6-(biotinylamino)-n-hexanoyl)- 829.4


aminobenzenesulfonyl)-(L)-prolyl-(L)-O-tert-


butyl-tyrosine


(224)N-(3,5-dichlorobenzenesulfonyl)-[3.1.0)-3-518


azabicyclohexane-2-carbonyl-(L)-4-


fluorophenylalanine


* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
AMPLE 225
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N-(3L5-Dichlorobenzenesulfonyl))-(L)-prolyl-(L)-3-(2-naphth~il)alanine.
Step A: ~,L)-3-(2-Naphthyl)alanine tent-butvl ester, hydrochloride
To a solution of N-Boc-2-naphthylalanine (1.0 g, 3.17
mmol) in a mixture of methylene chloride (7 mL) and cyclohexane (14
mL) were added t-butyl trichloroacetimidate (0.60 mL, 3.35 mmol)
and boron trifluoride-etherate (60 N,L, 0.473 mmol). The reaction
mixture was stirred for 5 hours at room temperature under a
nitrogen atmosphere and then treated a second time with the same
amounts of t-butyl trichloroacetimidate and boron trifluoride-etherate
as above. After stirring overnight, the mixture was filtered and the
filtrate evaporated. The product was obtained pure by silica gel
chromatography eluting with 10% diethyl ether in hexane; yield 843
mg. The product was treated with 1M HCl in ethyl acetate ( 11.5 mL)
for 18 hours at room temperature. The mixture was evaporated and
coevaporated several times with diethyl ether to afford the title
compound; yield 670 mg.
400 MHz 1H NMR (CD30D): 81.38 (s, 9H); 3.29-3.46 (m, 2H); 4.28 (t,
1H); 7.40-7.90 (m, 7H).
Step B: N-(Benzyloxvcarbonyl)-(L)-prolvl-(L)-3-(2-
naphthyl)alanine. tert-butyl ester.
To a solution of N-(benzyloxycarbonyl)-(~-proline (536 mg,
2.15 mmol) in methylene chloride (25 mL) were added 1-
hydroxybenzotriazole (434 mg, 3.21 mmol), N-methylmorpholine
(0.353 mL, 3.21 mmol), and (~-2-naphthylalanine tert-butyl ester
hydrochloride (660 mg, 2.14 mmol). After cooling in an ice-bath for 5
minutes, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide
hydrochloride (EDC) (493 mg, 2.57 mmol) was added. After 15
minutes, the cooling bath was removed and the mixture stirred
overnight under a nitrogen atmosphere. The mixture was diluted
with methylene chloride, washed with water, 2N HCl, saturated
NaHC03 solution, saturated brine solution, dried (anhydrous
magnesium sulfate), and evaporated. Silica gel chromatography
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eluting with 30% ethyl acetate in hexane afforded pure title
compound; yield 877 mg (81%).
Step C: (L)-Prolvl-(L)-3-(2-naphthyl)alanine tart butyl ester
A solution of ~T-(benzyloxycarbonyl)-(~-prolyl-(~-2-
naphthylalanine tart-butyl ester (870 mg, 1.73 mmol) in methanol (30
mL) was hydrogenated under an atmosphere of hydrogen gas in the
presence of 10% palladium-on-charcoal (75 mg) until complete
disappearance of starting material (several hours) as indicated by
TLC (30% ethyl acetate in hexane). The catalyst was removed by
filtration through Celite, the filter washed with methanol, and the
combined filtrate and washings evaporated to afford an oil that
crystallized upon standing; yield 604 mg (95%).
400 MHz 1H NMR (CD30D): 81.40 (s, 9H); 2.00 (m, 1H); 2.79 (m, 2H);
3.16 (dd, 1H); 3.58 (dd, 1H); 4.67 (dd, 1H); 7.32-7.81 (m, 7H).
Step D: N-(3.5-Dichlorobenzenesulfonvl))-(L) prolyl (L) 3 (2
nanhthyl)alanine tart-butyl ester
To a solution of (~)-prolyl-(~)-2-naphthylalanine tart-butyl
ester (400 mg, 1.09 mmol) in methylene chloride (10 mL) were added
N,N-diisopropylethylamine (470 p,L, 2.70 mmol), 4-
dimethylaminopyridine (13 mg, 0.106 mmol), and 3,5-
dichlorobenzenesulfonyl) chloride (320 mg, 1.30 mmol). The reaction
mixture was stirred for 2 hours at room temperature, diluted with
methylene chloride, washed with water, 2~T HCl, saturated NaHC03
solution, saturated brine solution, dried (Anhydrous magnesium
sulfate), and evaporated. Pure title compound was obtained by silica
gel chromatography eluting with 20% ethyl acetate in hexane; yield
501 mg (80%).
400 MHz 1H NMR (CD30D): 81.40 (s, 9H); 1.53-1.89 (m, 4H); 3.20-3.45
(m, 4H); 4.20 (dd, 1H); 4.69 (dd, 1H); 7.40-7.80 (m, lOH).
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Step E: N-(3,5-Dichlorobenzenesulfonyl))-(L)-prolyl-(L)-3- (2-
nanhthyl)alanine.
{224)A cooled solution of N-(3,5-dichlorobenzenesulfonyl))-(~-prolyl
(~-2-naphthylalanine tert-butyl ester (497 mg, 0.861 mmol) in
methylene chloride (25 mL) was treated with trifluoroacetic acid
(3.5 mL, 0.045 mol). The cooling bath was removed, and the
mixture was stirred until TLC (25% ethyl acetate in hexane)
indicated complete disappearance of starting material. The
reaction mixture was then evaporated, coevaporated with
methylene chloride (3X), toluene (2X), and finally methanol. The
product was dried under high vacuum; yield 445 mg (99%).
MS: m/e 521 (M); 537 (M + NH3)
400 MHz 1H NMR (CD30D): 81.51-1.87 (m, 4H); 3.19-3.46 (m, 4H); 4.20
(dd, 1H); 4.80 (dd, 1H); 7.39-7.82 (m, 10H).
The following compounds were prepared by the
procedures described in Example 225 using the appropriate amino
acid derivatives and acyl or sulfonyl chloride or alkyl or aryl
isocyanate:
Ex. Compound Name MS
(226) N-[4-(N'-2-toluylureido)phenylacetyl-(L)-prolyl- 495
(L)-norleucine
(227) N-(3,4-dimethoxybenzoyl)-(L)-prolyl-(L)- 393
norleucine
(228) N-(3,4-dimethoxybenzenesulfonyl))-(L)-pipecolyl- 516
(L)-tryptophan
(229) N-{4-nitrobenzenesulfonyl))-(L)-prolyl-(L)- 414
norleucine
(230) N-[3,5-di(trifluoromethyl)benzenesulfonyl)]-(L)- 505
prolyl-(L)-norleucine
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(231) N-(3,5-dichlorobenzenesulfonyl))-(L)-prolyl-(L)- 437
norleucine


(232)N-(3-trifluoromethylbenzenesulfonyl))-(L)-prolyl-437


(L)-norleucine


(233)N-[4-(benzoylamino)benzenesulfonyl))-(L)-prolyl-488


(L)-norleucine


(234)N-(4-methoxy-3,5-dinitrobenzenesulfonyl)-(L)-488


prolyl-(L)-norleucine


(235)N-(3-chlorobenzenesulfonyl))-(L)-prolyl-(L)-402


norleucine


(236)N-(3-trifluoromethylbenzenesulfonyl))-{L)-prolyl-521


(L)-3-(2-naphthyl)alanine


(237)N-(3-nitrobenzenesulfonyl))-(L)-prolyl-(L)-414


norleucine


(238)N-(3-cyanobenzenesulfonyl))-(L)-prolyl-(L)-394


norleucine


(239)N-(3,5-dichlorobenzenesulfonyl))-(L)-prolyl-(L)-510


tryptophan


(2.4(1)N-(3-methylbenzenesulfonyl))-(L)-prolyl-(L)-383


norleucine


(241)N-(3,5-dichlorobenzenesulfonyl))-(L)-3(S)-methyl-535


prolyl-(L)-3-(2-naphthyl)alanine


(242)N-(3-chlorobenzenesulfonyl)-(L)-prolyl-(L)-3-(2-488


naphthyl)alanine


(243)N-(3-fluorobenzenesulfonyl))-(L)-prolyl-(L)-3-(2-471


naphthyl)alanine


(244)N-phenylacetyl-(L)-prolyl-(L)-3-(2- 431


naphthyl)alanine


(245)N-(3-phenylpropionyl)-(L)-prolyl-(L)-3-(2-~5


naphthyl)alanine


(246)N-(phenylaminocarbonyl)-(L)-prolyl-(L)-3-(2-432


naphthyl)alanine




CA 02291778 1999-11-22
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(247) N-(3,5-dichlorobenzenesulfonyl))-(L)-2-methyl- 535
prolyl-(L)-3-(2-naphthyl)-alanine


(248) N-(benzenesulfonyl)-(L)-prolyl-(L)-3-(2-453


naphthyl)alanine


(249) N-(4-N'-phenylureidobenzenesulfonyl)-(L)-prolyl-587


(L)-3-(2-naphthyl)alanine


(250) N-(3-fluorobenzenesulfonyl)-(L)-5,5-dimethyl-499


prolyl-(L)-3-(2-naphthyl)alanine


(251) N-(4-N'-(2-toluyl)ureidobenzenesulfonyl)-(L)-601


prolyl-(L)-3-(2-naphthyl)alanine


(252) N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-4-547


iodophenylalanine


(253) N-(4-N'-benzylureidobenzenesulfonyl)-(L)-prolyl-601


(L)-3-(2-naphthyl)alanine


(254) N-(phenyloxalyl)-(L)-prolyl-(L)-3-(2- 445


naphthyl)alanine


(255) N-(benzylaminocarbonyl)-(L)-prolyl-(L)-3-(2-445


naphthyl)alanine


(256) N-(3-fluorobenzenesulfonyl)-(L)-2(S)-methyl-470


prolyl-(L)-4-fluorophenylalanine


(257) N-(3,5-dichlorobenzenesulfonyl)-(L)-2{S)-methyl-520


prolyl-(L)-4-fluorophenylalanine


(258) N-{3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-565


phenylalaninamide-N-methylsulfonamide


(259) N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-628


prolyl-(L)-4-iodophenylalanine


(260) N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-261**


phenylalanine


(261) N-(3,5-dichlorobenzenesulfonyl)-(L)-5- 520


methylprolyl-(L)-4-fluorophenylalanine


(262) N-(3,5-dichlorobenzenesulfonyl)-3- 568


phenylazetidinylcarbonyl-(L)-4-
fluorophenylalanine


CA 02291778 1999-11-22
WO 98!53814 PCT/LTS98/10940
(263)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-529


allylprolyl-(L)-4-fluorophenylalanine


(264)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-


prolyl-(L)-phenylalanine


(265)N-(3-trifluoromethylbenzenesulfonyl)-(L)-2(S)-530


methyl-prolyl-(L)-4-nitro-phenylalanine


(266)N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-methyl-502.3


prolyl-(L)-4-fluorophenylalanine


(26?)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-509


prolyl-(L)-4-cyanophenylalanine


(268)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-545


prolyl-(L)-4-(aminocarbonyl)-phenylalanine


(269)N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-methyl-631.4


prolyl-(L)-4-(N-t-butoxycarbonylaminomethyl)-


phenylalanine


(270)N-(3,5-dichlorobenzenesulfonyl)-(L)-3(R)-methyl-514.3


prolyl-(L)-4-(aminomethyl)-phenylalanine


* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
** (M - 159: Nf SOzAr cleavage)
EXAMPLE 271
N-(3-Trifluoromethvlphenvlsulfonvl) (L) 2(S) methyl_prolyl (L) 4
acetaminophenylalaning
Step A: N-(3-trifluoromethvlDhenylsulfonyl)-(L) 2(S) methy~p~rolvl
(L)-4-aminophen~n amine, methyl ester.
To a solution of N-(3-trifluoromethylphenylsulfonyl)-(L)-
2(S)-methyl-prolyl-(L)-4-nitrophenylalanine, methyl ester (0.45 g, 0.85
mmol; prepared according to the methodology described in Example 225)
in methanol (40 mL) was added 10% palladium on carbon catalyst (50
mg) and the resulting black suspension was stirred under 1 atm of
hydrogen for 45 min. The reaction mixture was filtered (Whatman
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CA 02291778 1999-11-22
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syringless filter device) and rotoevaporated under high vacuum to an off
white solid (0.42 g, 99% yield) which was used in the following step
without further purification.
1H-NMR (400 MHz, CDC13): b 8.12 (s, 1H), 8.05 (d, 1H, J = 7.8 Hz),
7.81 (d, 1H, J = 7.7 Hz), 7.64 (t, 1H, J = ~7.9 Hz), 7.03 (d, 1H, J = 7.6 Hz),
6.97 (d, 2H, J = 8.4 Hz), 6.73 {d, 2H, J = 8.4), 4.76 {m, 1H), 3.75 (s, 3H),
3.48
(m, 1H), 3.28 (m, 1H), 3.14 (dd, 1H, J = 14.2, 5.4 Hz), 2.98 {dd, 1H, J =
14.2,
6.9 Hz), 2.29 (m, 1H), 1.78 (m, 1H), 1.62 (m, 2H), 1.57 (s, 3H).
Step B: N-(3-trifluoromethylphen ly sulfonyl)-(L)- 2(S)-methyl-prolvl-
(L)-4-acetaminophenvlalaning, methyl ester
To a solution of N-(3-trifluoromethylphenylsulfonyl)-(L)-
2(S)-methyl-prolyl-(L)-4-aminophenylalanine, methyl ester (42 mg, 0.082
mmol) in dry dichloromethane (0.5 mL) at 0 °C, was added successively
2,6-lutidine (0.03 mL, 0.25 mmol; 3.0 equiv), acetyl chloride (0.01 mL,
0.125 mmol; 1.5 equiv), and 4-dimethylaminopyridine ( 10 mg, 0.082
mmol; 1.0 equiv). The yellow reaction mixture was stirred overnight.
After this time, 1.0 N hydrochloric acid was added followed by extraction
with ethyl acetate (3x). The combined organic layer was successively
washed with saturated sodium bicarbonate solution and saturated salt
solution and dried over anhydrous magnesium sulfate. The mixture
was filtered and concentrated to furnish an orange-yellow oil (46 mg,
100% crude yield) which was purified by preparative thin layer
chromatography (80% ethyl acetate, 20% hexanes). Yield: 39 mg (85%).
1H-NMR (400 MHz, CDCl3): 8 8.11 (s, 1H), 8.04 (d, 1H, J = 8.0 Hz),
7.82 (d, 1H, J = 7.7 Hz), 7.64 (t, 1H, J = -7.9 Hz), 7.41 (d, 1H, J = 8.4 Hz},
7.25 (s, 1H), 7.09 (d, 2H, J = 8.4 Hz), 7.07 (d, 1H, J = ~8.0 Hz), 4.80 (m,
1H),
3.75 (s, 3H), 3.49 (m, 1H), 3.24 (m, 2H), 3.04 (dd, 1H, J =-.14.0, ~7.0 Hz),
2.29 (m, 1H), 2.13 (s, 3H), 1.75 (m, 1H), 1.61 (m, 2H), 1.57 (s, 3H).
Step C: N-(3-trifluoromethylphenylsulfonyl)-(L)- 2(S)-methyl~rol
(L)-4-acetaminoRhenvlalanine.
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To a solution of N-(3-trifluoromethyl)-2(S)-methyl-prolyl-4-
acetamino-(S)-phenylalanine, methyl ester (33 mg, 0.059 mmol) in
ethanol (1.0 mL) was added 0.2 N sodium hydroxide solution (0.60 mL,
0.12 mmol; 2.0 equiv). The reaction mixture was stirred overnight (15 h)
and then acidified with 1.0 N hydrochloric acid and extracted with ethyl
acetate (3x). The combined organic layer was washed with saturated
salt solution, dried over anhydrous magnesium sulfate, and
rotoevaporated to yield an off white solid (31 mg, 97% yield).
MS: m/e 542 (M+H+); 559 (M+NH4+).
1H-NMR (400 MHz, CD30D): 8 8.08 (m, 2H), 7.95 (d, 1H, J = 7.7 Hz),
7.76 (t, 1H, J = ~7.9 Hz), 7.48 (m, 3H), 7.18 (d, 2H, J = 8.4), 4.69 (m, 1H),
3.43 (m, 1H), 3.32 (m, 2H), 3.05 (dd, 1H, J = 14.0, -7.0 Hz), 2.12 (m, 1H),
2.08 (s, 3H), 1.71 (m, 3H), 1.56 (s, 3H).
The following compounds were prepared by the
procedures described in Example 271 using the acyl or sulfonyl
chloride or alkyl or aryl isocyanate:
Ex. Compound Name MS
(272) N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)- 633
methyl-prolyl-(L)-4-(N'-(2-
toluyl)ureido)phenylalanine.
(273) N-(3-trifluoromethylphenylsulfonyl)-(L)-2{S)- 718
methyl-prolyl-(L)-4-(N'-(4'-
fluorophenylsulfonyl)ureido)phenylalanine.
(274) N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)- 572
methyl-prolyl-(L)-4-
(ethoxycarbonyl)aminophenylalanine.
(275) N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)- 766
methyl-prolyl-(L)-4-(4'-(N'-(2-
toluyl)ureido)phenylacetyl)aminophenylalanine.
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(276)N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-658


methyl-prolyl-(L)-4-(4'-


fluorophenylsulfonyl)aminophenylalanine.


(277)N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-618


methyl-prolyl-(L)-4-


(phenylacetyl)aminophenylalanine.


(278)N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-622


methyl-prolyl-(L)-4-(4'-


fluorobenzoyl)aminophenylalanine.


(279)N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-600


methyl-prolyl-(L)-4-


(isobutyloxycarbonyl)aminophenylalanine.


(280)N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)-578


methyl-prolyl-(L)-4-
methylsulfonylaminophenylalanine.
(281) N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)- 637
methyl-prolyl-(L)-4-(N'-(4-
fluorophenyl)ureido)phenylalanine.
(282) N-(3-trifluoromethylbenzenesulfonyl)-(L)-2(S)- 621
methyl-prolyl-(L)-4-(N-( 1,1-dioxo-1,2-
isothiazolidinyl)-phenylalanine
(283) N-(3-trifluoromethylphenylsulfonyl)-(L)-2(S)- 585
methyl-prolyl-(L)-4-(N'-(4-(2-oxo-1-pyrrolidinyl)-
phenylalanine.
* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
EXAMPLE 284
N-(3.5-dichorobenzenesulfonyl)-(L)-prolyl-(L)-4-(4'-
fluorobenzoyl)phenylalanine
Step A: 4-Iodo-(L)-Phenylalanine, tert-butyl ester hydrochloride
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To a suspension of N-Boc-4-iodo-(~-phenylalanine (1.0 g, 2.56
mmol) in methylene chloride (7 mL) and cyclohexane (14 mL) were
added t-butyl trichloroacetimidate (0.48 mL, 2.68 mmol) and boron
trifluoride-etherate (48 p,L). The reaction mixture was stirred for 5 hours
at room temperature under a nitrogen atmosphere and then treated a
second time with the same amounts of t-butyl trichloroacetimidate and
boron trifluoride-etherate as above. After stirring overnight, a third
addition was made, and the mixture was stirred a further 3 hours. The
mixture was then filtered and the filtrate evaporated. The product was
obtained pure by silica gel chromatography eluting with 10% diethyl
ether in hexane; yield 650 mg. The product was treated with IM HCl in
ethyl acetate (7.3 mL) for 18 hours at room temperature. The mixture
was evaporated and coevaporated several times with diethyl ether to
afford the title compound; yield 522 mg.
400 MHz IH NMR (CD30D): 81.42 (s, 9H); 3.13 (d, 2H); 4.18 (t, 1H); 7.09
(d, 2H); 7.75 (d, 2H).
Step B: N-(3 5-Dichlorobenzenesulfonvl (L) proline
To a mixture of (~-proline methyl ester hydrochloride (838
mg, 5.06 mmol) in methylene chloride (25 mL) at 0°C were added N.N-
diisopropylethylamine (2.64 mL, 15.2 mmol) and a solution of 3,5-
dichlorobenzenesulfonyl chloride (1.49 g, 6.07 mmol) in methylene
chloride (5 mL). The cooling bath was removed, and the mixture was
stirred overnight at room temperature. It was then diluted with
methylene chloride, washed with 1~T hydrochloric acid, saturated
NaHC03, saturated brine solution, dried over anhydrous sodium
sulfate, and evaporated. The methyl ester was obtained pure by silica gel
chromatography eluting with 10% acetone in hexane; yield 1.49 g. It
was then taken up in ethanol (50 mL) and treated with 0.2 N sodium
hydroxide (26.6 mL) for 1.5 hours at room temperature. The mixture
was acidified with glacial acetic acid, concentrated, the residue taken up
in methylene chloride, washed with water, saturated brine solution,
dried (Na2S04), and evaporated to give the title compound; yield 1.4 g.
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400 MHz 1H NMR (CD30D): 81.80-2.15 (m, 4H); 3.35-4.45 (m, 2H); 4.30
(dd, 1H); 7.76 (m, 1H); 7.83 (m, 2H).
Step C: N-(3,5-Dichlorobenzenesulfony] )-l (L)-prolyl-(L)-4-
iodophenylalanine, tent-butyl ester.
To a solution of ~T-(3,5-dichlorobenzenesulfonyl)-(~-proline
(386 mg, 1.19 mmol) in methylene chloride (23 mL) were added 1-
hydroxybenzotriazole (241 mg, 1.79 mmol), N-methylmorpholine (0.33
mL, 2.98 mmol), and 4-iodo-(~-phenylalanine tent-butyl ester
hydrochloride (458 mg, 1.19 mmol). After cooling in an ice-bath for 5
minutes, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
(EDC) (274 mg, 1.43 mmol) was added. After 15 minutes, the cooling
bath was removed, and the mixture was stirred overnight under a
nitrogen atmosphere. The mixture was diluted with methylene
chloride, washed with water, 1N HCl, saturated NaHC03 solution,
saturated brine solution, dried (Anhydrous magnesium sulfate), and
evaporated. Silica gel chromatography eluting with 20% ethyl acetate in
hexane afforded pure title compound; yield 651 mg (84%).
MS: m/e 653 (M + 1)
400 MHz 1H NMR (CD30D): 81.45 (s, 9H); 1.65-1.85 (m, 4H); 3.0 (dd, 1H);
3.13 (dd, 1H); 3.45 (m, 1H); 4.20 (m, 1H); 4.55 (dd, 1H); 7.05 (d, 2H); 7.64
(d,
2H); 7.80 (s, 3H).
Step D: N-(3.5-dichlorobenzenesulfonvl)-(L)- r~olvl-(L)-4-(4'-
fluorobenzo~Rhenylalanine, tert-bull ester.
A solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-4-
iodo-(L)- phenylalanine tert-butyl ester(100 mg, 0.15 mmol), 4-
fluorobenzeneboronic acid (23 mg, O.lfi mmol), potassium carbonate(62
mg, 0.45 mmol), bis(triphenylphosphine)-palladium(II) chloride (4 mg,
0.0057 mmol) in anisole(4 ml) was flushed with nitrogen, then flushed
with CO, and a balloon of CO was attached. The solution was then
stirred at 80°C for 5 hours on a timer overnight. The following day the
solution was diluted with methylene chloride, washed once with water,
_g0_


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once with brine, dried over Anhydrous magnesium sulfate, and solvent
removed in vacuo. The desired product was obtained by silica gel
chromatography eluting with methylene chloride, followed by 10% ethyl
acetate in methylene chloride; yield 70 mg (72%)
MS: m/e 666.2 (M+H+NH3)
400 MHz 1H NMR (CD30D): S 1.46(s,9H); 1.65-1.95(m,4H); 3.05-3.15
(dd,lH); 3.47(m,lH); 4.2(dd,lH); 4.65(m,lH); 7.20(t,2H); 7.45(d,2H);
7.70(d,2H);7.76-7.85(m,SH)
P N-(3 5-dichoroben Pnpanlf
Ste E: ~~ onvl) fL) nrnl~~1 (~) 4 (4'
fluorobenzovl)p~ anin
A solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-
4-(4-fluorobenzoyl)phenylalanine, tert-butyl ester (23 mg, 0.035 mmol) in
methylene chloride(L2 mL) was cooled in ice bath. Trifluoroacetic acid
(0.167 mL, 2.17 mmol) was then added, and ice bath was removed and
reaction mixture was allowed to stir overnight at room temperature.
The reaction mixture was then evaporated, coevaporated with methylene
chloride(2X), toluene(2X), and methanol(2X). The product was obtained
pure by eluting with 20% ethyl acetate in methylene chloride, followed by
8% methanol in methylene chloride; yield 19 mg{91%)
MS: m/e 609.8(M+H+NH3)
400 Mhz 1H NMR (CDsOD): 81.6-1.95(m,4H): 3.1-3.45(m,4H): 4.17 (dd,lH):
4.55(m,lH): 7.2(t,2H): 7.4(d,2H): 7.66(d,2H); 7.78-7.85(m,SH)
The following compounds were prepared by the
procedures described in Example 284 using the appropriate
arylboronic acid derivative in Step D:
Compound Name MS
(285) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4'- 604.8
(2-methoxybenzoyl)phenylalanine
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(286) N-{3,5-dichorobenzenesulfonyl)-(L)-2(S)-methyl- 624
prolyl-(L)-4-(4'-fluorobenzoyl)phenylalanine
* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
EXAMPLE 287
N-(3.5-dichlorobenzenesulfonyl)-(L)-prolvl-(L)-4-(4-fluorobenz,~phenXl
alanine
Step A: N-(3.5-dichlorobenzenesulfonvl)-(L)-pro13r1-(L)-4-
(4-fluoro-a-hvdroxvbenz 1)~phenvlalanine tert-but 1 ester
A solution of N-{3,5-dichorobenzenesulfonyl)-(L)-prolyl-(L)-4-
(4'-fluorobenzoyl)phenylalanine (38 mg) in methanol (5 mL) was cooled
to 0° C. Sodium borohydride (3 mg) was added. After stirring for 20
min,
the solvent was removed by rotoevaporation and the residue dissolved in
dichloromethane (30 mL). The solution was successively washed with
water and saturated salt solution and dried over anhydrous magnesium
sulfate. The mixture was filtered and the solvent was removed by
rotoevaporation. The title compound (38 mg) was recovered and used
with no further purification in the subsequent reaction.
Step B: N-(3,5-dichlorobenzenesulfon lv )-(L)=prolyl-(L)-4
(4-fluorobenzyl)phe ~lalanine
A solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-
(4-fluorophenyl-hydroxymethyl)phenylalanine, tert-butyl ester (38 mg)
and triethylsilane (21 N.L) in anhydrous dichloromethane was flushed
with dry nitrogen for five minutes. The solution was then cooled in an
ice bath and boron trifluoride etherate (16 uL) was added. After stirring
for 3 hours, methanol (1 mL) was added and the solvent was removed by
rotoevaporation. The residue was dissolved in ethyl acetate and the
solution successively washed with saturated sodium bicarbonate
solution and saturated salt solution and then dried over anhydrous
magnesium sulfate. After the mixture was filtered, the solvent was
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removed by rotoevaporation. The residue was purified by flash column
chromatography on silica gel eluted with 97.75% dichloromethante, 2%
methanol and 0.25% acetic acid to yield the title compound (14 mg).
M/S: m/e = 597.2 (M + NH4).
1H NMR (400 MHz, CD30D): 81.5-1.7 (m, 2H), 1.75-1.82 (m, 2H), 2.95-3.05
(m, 1H), 3.2-3.4 (m, 3H), 3.88 (s, 2H), 4.1-4.2 (m, 1 H), 4.6-4.7 (m, 1H),
6.90
(t, J= 9, 2H), 7.1-7.22 (m, 6H), 7.72 (s, 2H), 7.76 (s, IH).
The following compounds were prepared by the
procedures described in Example 287:
Ex. Compound Name MS *
(288) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4- 608.3
(2-methoxybenzyl)phenylalanine
* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
I5 ABLE 289
N-(3.5-Dichlorobenzenesulfon ly )-(L)-pro yl-(L) 4 (2 nitrophenoxy~
,nhenylalanine
Step A: N-Boc-4-(2-nitro he_n_oxv) (L~uhenvlalaninea meth3il ester
To a solution of N-Boc-(L)-tyrosine, methyl ester (500 mg)
and potassium carbonate (467 mg) in dimethylformamide (5 mL) was
added dropwise 1-fluoro-2-nitrobenzene (189 uL). The yellow solution
was stirred for 3 days at room temperature. The mixture was diluted
with ether which was subsequently washed with 1N hydrochloric acid,
water, saturated salt solution and dried over anhydrous magnesium
sulfate. After filtration, the solvent was removed by rotoevaporation to
yield the title compound (?00 mg) which was used in the subsequent
reaction without further purification.


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1H NMR (400 MHz, CD30D): 81.38 (s, 9H), 3.85-3.15 (m, 2 H), 4.3-4.4(m,
1H), 6.95-7.1 (m, 3H), 7.24-7.3(m, 3H), 7.55-7.61 (t, 1H), 7.97-7.97(m, 1H).
Step B: 4-(2-nitronhenoxv)-(L)-~ylalanine. methyl ester
h,~idrochloride
N-Boc-4-(2-nitrophenoxy)-(L)-phenylalanine, methyl ester (600
mg) was stirred in a solution of 1N hydrochloric acid in ethyl acetate (10
mL) for 18 hours at room temperature. A precipitate formed, the solvent
was removed by rotoevaporation, and co-evaporated with Et20 (2x). The
solid was than suspended with ethyl acetate, filtered, washed with
diethyl ether, and allowed to air dry. The title compound was recovered
(490 mg) and used in the subsequent reaction without further
purification.
I5 Step C: N-(3.5-Dichlorobenzenesulfonvl)-(L)-prolvl-(L)-4-(2-
nitrophenoxv)phenvlalanine. methyl ester.
A solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-proline (429
mg), 4-(2-nitrophenoxy)-(L)-phenylalanine, methyl ester hydrochloride
(445 mg), I-hydroxybenztriazole (255 mg), N-methylmorpholine (0.35 mL)
in dichloromethane (32 mL) was cooled to 0 °C. 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC; 289 mg)
was then added. The reaction was allowed to warm to room
temperature and stirred for 17 hr. The reaction was diluted with
dichloromethane ( 100 mL) and successively washed with water, 1N
hydrochloric acid, saturated sodium bicarbonate solution, and saturated
salt solution. The organic layer was dried over anhydrous magnesium
sulfate. The solution was filtered and the solvent removed by
rotoevaporation. The residue was purified by flash column
chromatography on silica gel eluted with 20% ethyl acetate in hexane to
afford the title compound (714 mg) which was used in the subsequent
reaction.


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Step D: N-(3 5-Dichlorobenzenesul nvl) (L) ~rolvl (L) 4 (2
~mtrophenoxy)~henvlalanin~
N-(3,5-Dichlorobenzenesulfonyl)-(L)-prolyl-(L)-
4-(2-vitro-phenoxy)-phenylalanine, methyl ester (110 mg) was dissolved
in ethanol (6 mL) and a solution of potassium hydroxide (15 mg) in water
(2 mL) was added. After stirring for 20 minutes, the reaction was
acidified with acetic acid and the solvent removed by rotoevaporation.
The residue was dissolved in ethyl acetate (40 mL), and the solution
successively washed with saturated sodium bicarbonate solution and
saturated salt solution. The solution was dried over anhydrouus
magnesium sulfate, then filtered and the solvent removed by
rotoevaporation to afford the title compound (40 mg).
M/S: m/e 625(M+NH4>+.
1H NMR (400 MHz, CDsOD): S 1.63-1.72(m, 1H), 1.75-2.92(m, 3H), 3.01-
3.08(dd, 1H), 3.25-3.35(m, 2H), 3.4-3.5 (m, 1H), 4.19 (dd, J= 6,1, 1H), 4.68-
4.74 (m, 1H), 6.97-7.05 (m, 3H), 7.2-7.35 (m, 3H), 7.45-7.5 (m, 1H), 7.77 (s,
3H), 7.91 (dd, J= 7,2, 1H).
The following compound was prepared by the procedures
described in Example 289:
Example Compound Name MS*
(290) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4- 625
(4-nitrophenoxy)-phenylalanine
(291) N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl- 639
prolyl-(L)-4-(2-nitrophenoxy)-phenylalanine
* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
~XAMI°LE 292
N-(3.5-Dichlo_robenzenesulfonvl -(L)-Dro~yl (L) 4 (2 amino henoxv)
phenylalanine
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Step A: N-(3.5-Dichlorobenzenesulfonvl)-(L)-prolvl-(L)-4-(2-
aminophenoxy)-phenylalanine~, methyl ester
To a solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-4-(2-vitro-phenoxy)-phenylalanine, methyl ester (120 mg) in ethanol
(4.5 mL) was added iron filings (42 mg) and acetic acid (0.5 mL).
Reaction was refluxed for 3 h then cooled to room temperature. The
mixture was filtered through a pad of celite and the solvent was removed
by rotoevaporation. The resultant tar was dissolved in ethyl acetate and
successively washed with saturated sodium bicarbonate solution and
saturated salt solution. The organic layer was dried over anhydrous
magnesium sulfate, filtered and the solvent removed by rotoevaporation.
The resiude was purified by flash column chromatography on silica gel
eluted with 40% ethyl acetate in hexane to afford N-(3,5-
dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-aminophenoxy)-
phenylalanine, methyl ester (75 mg) which was used in the subsequent
reaction.
Step B: N-.(3.5-Dichlorobenzenesulfon ly )-(L)=prolvl-f L)-4-(2-
aminophenoxy)-~ylalanine
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-
aminophenoxy)-phenylalanine, methyl ester was hydrolyzed by the
procedure in Example 289, step D to afford N-(3,5-
dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-aminophenoxy)-
phenylalanine.
M/S: m/e 578(M+1).
1H NMR (400 MHz, CD30D): 81.62-1.9 (m, 4H), 3.0-3.07 (dd, 1H), 3.2-
3.3(m, 2H), 3.4-3.5 (m, 1H), 4.19 (t, 1H), 4.62-4.7 (m, 1H), 6.6-6.65 (m, 1H),
6.73-6.77 (dd, 1H), 6.85-6.95 (m, 4H), 7.2 (d, J=2, 2H), 7.78 (s, 3H), 8.1-
8.15
(d, 1H).
EXAMPLE 293


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N- -Di r z sulfo 1 - r 1- L -4- cet min h no
phenvlalanine
Step A: N-(3 5-DichlornhPn~PrPQ"1 0~~ l~nrol3il (L) 4 (2
acetvlaminophenoxv)-lalaning r,pthyl ester
To a solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-4-(2-amino-phenoxy)-phenylalanine, methyl ester (55 mg) in pyridine
(0.31 mL) and dichloromethane (4 mL) was dropwise added acetic
anhydride (0.16 mL). After stirring for 1 hr, the reaction was diluted
with dichloromethane (50 mL) and successivley washed with water and
saturated salt solution. The solution was dried over anhydrous
magnesium sulfate, filtered and the solvent removed by rotoevaporation.
The residue was purified by flash column chromatography on silica gel
eluted with 5% ethyl acetate in dichloromethane to afford N-(3,5-
dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-acetylaminophenoxy)-
phenylalanine, methyl ester (41 mg) which was used in the subsequent
reaction.
Step B: N-(3.5-Dichlorobenzenesulfonvl)-(L) ;prolyl (L) 4 (2
~vlaminophe~p~ylalanine
N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4-(2-
acetylaminophenoxy)-phenylalanine, methyl ester was hydrolyzed by the
procedure in Example 289, step D to afford N-(3,5-
dichlorobenzenesulfonyl)-(L)-proiyl-(L)-4-(2-acetylaminophenoxy)-
phenylalanine.
M/S: m/e 637(M+NH4)+.
1H NMR (400 MHz, CDsOD): d 1.6-1.95 (m, 4H), 2.06 (s, 3H), 3.0-3.08 (dd,
1H), 3.2-3.3 (m, 2H), 3.4-3.48 (m, 1H), 4.15-4.2 (m, 1H), 5.55-5.61 (m, 1H),
6.8-6.85 (d, 1H), 6.91 (d, J= 9, 2H), 6.98-7.08 (m, 2H), 7.26 (d, J=9, 2H),
7.78
(s, 3H), 8.85-8.90 (dd, 1H).
The following compounds were prepared by the
procedures described in Example 293:
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Ex. Compound Name MS*
(294) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-4- 637
(4-acetylaminophenoxy)-phenylalanine
(295) N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)- 636
methylprolyl-(L)-4-(2-acetylaminophenoxy)-
phenylalanine
* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
EXAMPLE 296
N-(3,5-Dichlorobenzenesulfonyl)-2-(S)-meth 1-~prolvl-4-
(2-c~ hp enox~phenylalanine
Step A: N-Boc-4-(2-cyanophenoxy)-nhenylalanine methyl ester
A solution of 500 mg of N-Boc-4-(L)-tyrosine, methyl ester,
205 mg 2-fluorobenzonitrile, 245 mg KF 40 wt% on alumina, 45 mg 18-
crown-6, and 7 mL of acetonitrile was run at reflux for seven days. The
reaction was then diluted with methylene chloride, and washed with
water and saturated salt solution. The organic layers were then dried
over anhydrous magnesium sulfate and the solvent was removed in
vacuo. The product was purified via silica gel chromatography eluted
with 80% hexane:20% acetone to yield 253 mg of the product.
1H NMR (400 Mhz, CD30D): 81.38(s, 9H), 2.9(dd, 1H), 3.13(dd,1H), 3.70(s,
3H), 3.38(m, 1H), 6.88(d, 1H), 7.03 (d, J=9, 2H), 7.2(t, 1H), 7.29(d, J=9,
2H),
7.55(t, 1H), 7.72,(d, 1H).
Step B: 4-(2-cyanophenox~phenvlalanin~,, methyl
ester.hvdrochloride
The reaction was performed by an analogous procedure as
described in Example 289, step B to yield the title compound.
_6g_


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Step C: N-Boc-2-(S)-methyl-(, L)~nrollvl-4-(2-cyanophenox,~
~henylalanine methyl ester
To a solution of 131 mg of N-Boc-2-(S)-methyl-(L)-proline, 190
mg 4-(2-cyanophenoxy)-phenylalanine, methyl ester hydrochloride, 297
mg PyBOP, and 4 mL of methylene chloride at 0° C was added 300 uL of
diisopropylethylamine via syringe. The reactants were allowed to warm
to room temperature and said reaction was run over the weekend. The
reaction was then diluted with methylene chloride, washed with water,
1N hydrochloric acid, saturated sodium bicarbonate solution, and
saturated salt solution. The organic layer was dried over anhydrous
magnesium sulfate and concentrated in vacuo. The product was
purified via silica gel chromatography, eluted with 80% hexane:20%
acetone to yield 263 mg of the title compound.
Step D: N-Boc-2-(S)-meth 1-pro 4 (2 cyanophen
phenvlalanine, methyl ester hydrochloride
The reaction was performed by an analogous procedure as
described in Example 289, step B to yield the title compound.
Step E: N-(3.5-Dichlorobenzenesulfonvl)-2-(S)-methyl (L) rol~l_
4-(2-cvano henoxv)-phenvlalanine, methyl ester
To a solution of 95 mg of N-Boc-2-(S)-methyl-(L)-prolyl-4-(2-
cyanophenoxy)-phenylalanine, hydrochloride, 61 mg 3,5-
dichlorobenzenesulfonyl chloride, and 2.5 mL of tetrahydrofuran at 0° C
was added 110 uL of diisopropylethylamine via syringe. The reaction
was allowed to warm to room temperature and run at said temperature
overnight. The reaction was diluted with methylene chloride, washed
with water, 1N hydrochloric acid, saturated sodium bicarbonate
solution, and saturated salt solution.. The organic layer was dried over
anhydrous magnesium sulfate and concentrated in vacuo. The produt
was purified via silica gel chromatography, eluted with 80% hexane:20%
acetone to yield 62 mg of of N-(3,5-dichlorobenzenesulfonyl)-2-(S)-methyl-
(L)-prolyl-4-(2-cyanophenoxy)-phenylalanine, methyl ester.
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Step F: N-(3,5-Dichlorobenzenesulfonyl)-2-(S)-methyl-(L)-prolyl- 4-
(2-cvanophenox~phenylalanine
To a solution of 62 mg of N-(3,5-dichlorobenzenesulfonyl)-2-
(S)-methyl-(L)-prolyl-4-(2-cyanophenoxy)-phenylalanine, methyl ester in
5 mL of ethanol was added a solution of I1 mg potassium hydroxide in 2
mL of water. After 1.5 hours the solvent was removed in vacuo. The
resultant solid was then dissolve in methylene chloride and washed with
0.5 M hydrochloric acid and saturated salt solution. The organic layers
were dried over anhydrous magnesium sulfate and concentrated in
vacuo. The formed diastereomers were separated via HPLC using a
YMC ODS-AQ column, eluting with 80% MeOH: 20% WATER + 0.1%
TFA. The faster eluting product was shown to be the desired product.
M/S: m/e 619 (M+1+NH3).
1H NMR (400 Mhz, CD30D): 81.60(s, 3H), 1.7-1.9(m, 3H), 2.12-2.21(m,
1H), 3.08-3.16(dd, 1H), 3.3-3.5(m), 4.65-4.75(m, 1H), 6.91(d, J=8 1H), 7.04(d,
2H), 7.15 (t, 1H), 7.36 (d, J=9, 2H), 7.4-7.5 (t, 1H), 7.6-7.8(m, 4H).
The following compound was prepared by the procedures
described in Example 296:
Ex. Compound Name MS*
(297) N-(3,5-Dichlorobenzenesulfonyl)-2-(S)-methyl-(L)- 619
prolyl-4-(4-cyanophenoxy)-phenylalanine
EXAMPLE 298
N-(3 5-Dichlorobenzenesulfo~yl)-(L)-prolyl-(L)-O-tent-butyl-tyrosine
Step A: N-(3 5-Dichlorobenzenesulfonvl)-(L)-prolvl-(L)-O-tert-
but~vrosine. methyl ester.
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To a solution of 3,5-dichlorobenzenesulfonyl-(L)-proline
(from Example 284, Step B) (1.70 gm, 5.23 mmole) in dry
dichloromethane (15 mL) was added 1-hydroxybenzotriazole hydrate
(782.3 mg, 5.78 mmole) followed by N-methylmorpholine (1.45mL, 13.1
mmole), (L)-O-tent-butyl-tyrosine, methyl ester hydrochloride (1.58
gm, 6.31 mmole), and 1-ethyl-3-(3-dimethylamino-propyl)
carbodiimde (1.41 gm, ?.36 mmole). Additional dichloromethane (5
mL) was added and the solution stirred under nitrogen at 25°C
overnight. Water was added and the layers separated. The aqueous
layer was extracted with ethyl acetate (3 x 15 mL). The combined
organic layers were successively washed with water (2 x 20 mL) and
saturated salt solution and dried over anhydrous magnesium sulfate.
After filtration, the solvent was removed by rotoevaporation. The
residue was purified by flash column chromatography on silica gel
eluted with 5-35% ethyl acetate in hexanes to yield N-(3,5-
dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-tert-butyl-tyrosine, methyl
ester as a pale white foam (2.85 gm, 98% yield).
MS: m/e 557.4 (M+1)+.
400 MHz 1H NMR (CD30D): 81.28 (s, 9H),1.49-1.66 (m, 3H), 2.03-2.07
(m, 1H), 2.99 (dd, J = 14.0, 7.5 Hz, 1H), 3.06-3.12 (m, 1H), 3.19 (dd, J =
14.1, 5.5 Hz, 1H), 3.34-3.39 (m, 1H), 3.74 (s, 3H), 4.04-4.07 (m, 1H), 4.76-
4.81 (m, 1H), 6.88 (d, J = 8.4 Hz, 2H), 6.99 (d, J = 8.4 Hz, 3H), 7.58 (t, J =
1.8 Hz, 1H), 7.69 (d, J = 1.8 Hz, 2H).
Step B: N-(3.5-DichlorobenzenPsulfon ly )-(L)-~Yl ('L~ O tent
but ~~l-~yrosine.
Under a dry nitrogen atmosphere, to a solution of 1.20gm
(2.15 mmole) of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-tert-
butyl-tyrosine, methyl ester ( 1.20 gm, 2.15 mmole) in dry ethanol
(25.8mL) was added dropwise an aqueous 0.2N sodium hydroxide
solution (12.9mL, 2.58 mmole). The reaction was stirred for 1.5 hr at
room temperature. A 1.OM aqueous solution of acetic acid (~2 mL)
was added until pH 4-5 was obtained. The solvent was removed by
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rotoevaporation and the residue dissolved in dichloromethane and
water. The layers were separated and the aqueous layer was
extracted with dichloromethane (3 x 20 mL). The organic layers were
combined, and successively washed with water, saturated salt
solution, and dried over anhydrous sodium sulfate. After filtration,
the solvent was removed by rotoevaporation. The residue dissolved in
a minimum of dichloromethane and purified on a 4000 ~,m silica gel
plate on a Chromatotron, eluted with 1-10% methanol in
dichloromethane to yield N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-O-tert-butyl-tyrosine as a pale yellow foam ( 1.15 gm, 99% yield).
MS: m/e 543.3 (M+1)+.
400 MHz NMR (CD30D) 81.28 (s, 9H), 1.60-1.69 (m, 1H), 1.70-1.79 (m,
1H), 1.82-1.89 (m, 2H), 3.02-3.06 (m, 1H), 3.21-3.30 (m, 4H), 3.41-3.49
(m, 1H), 4.19 (br t, J = 6.60 Hz, 1H), 4.62 (br s, 1H), 6.90 (d, J = 8.3 Hz,
2H), 7.18 (d, J = 8.4 Hz, 2H), 7.78 (s, 3H).
EXAMPLE 299
N-(3~5-Dichlorobenzenesulfonvl)-(L)-prolyl-(L)-O-methyl-tyrosine.
Step A: N-(3.5-Dichlorobenzenesulfonyl)-(L)-prolvl-(L)-O-tert-
butyl-tyrosine, tert-butyl ester
By the procedure of Example 284, step C, N-(3,5-
dichlorobenzenesulfonyl)-(L)-proline was coupled with (L)-O-tert-
butyl-tyrosine, tert-butyl ester hydrochloride. The product was
purified by flash column chromatography on silica gel eluted with
5-35% ethyl acetate in hexane and isolated as a white foam (85%
yield).
MS: m/e 599.0 (M+1)+.
400 Mhz 'H NMR (CDCl3) b 1.28 (s, 9H), 1.42 (s, 9H), 1.56-1.63 (m, 4H),
2.05-2.08 (m, 1H), 2.99 (dd, J = 14.0, 6.7 Hz, 1H), 3.09-3.17 (m, 2H), 3.35-
3.38 (m, 1H), 4.06-4.08 (m, 1H), 4.67 (br dd, J = 14.0, 6.3 Hz, 1H), 6.87
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(br d, J = 8.5 Hz, 2H), 7.03 (br d, J = 8.4 Hz, 3H), 7.06 (br d, J = 7.6 Hz,
1H), 7.57 (t, J = 1.8 Hz, 1H), 7.70 (d, J = 1.8 Hz, 2H).
Step B: N-(3 5-Dichlorobenzenesulfonyl) (L)~rolvl (L) t~rosine~
tart-butyl ester
To a solution of N-(3,5-dichlorobenzenesulfonyl)-{L)-
prolyl-(L)-O-tart-butyl-tyrosine, tent-butyl ester (1.20 gm, 2.00 mmole)
in dry dichloromethane (6 mL) at 0° C under a dry nitrogen
atmosphere was dropwise added a 50% v/v solution of trifluoroacetic
acid in dichloromethane (3.08 mL, 20 mmol) over a 10 min period.
After stirring for 2 hr, the reaction mixture was quenched at 0° C
with an aqueous 5% sodium bicarbonate solution to pH = 7-8. The
layers were separated and the organic layer dried over anhydrous
magnesium sulfate. After filtration, the solvent was removed by
rotoevaporation and the residue purified by flash column
chromatography on silica gel eluted with 1-10% methanol in
dichloromethane to yield N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-tyrosine, tart-butyl ester as a white foam (1.71 gm, 78% yield).
MS: m/s 543.4 (M+1)+.
400 MHz 1H NMR (CDCls) $ 1.45 {s, 9H), 1.55-1.63 (m, 3H), 2.07 (m,
1H), 2.94 (dd, J = 14.1, 6.90 Hz, 1H), 3.09-3.16 (m, 2H), 3.37-3.39 (m,
1H), 4.06-4.09 (m, 1H), 4.65-4.?0 (m, 1H), 6.71 (d, J = 8.5 Hz, 2H), 7.01
(d, J = 8.5 Hz, 2H), 7.06 (d, J = 7.7 Hz, 1H), 7.58 (t, J =1.8 Hz, 1H), ?.70
(d, J =1.8 Hz, 2H).
Step C: N-(3 5-dichlorobenzenesulfonyl) (L) ~rolvl (L) O meth
tyrosine tent-butyl ester
To a solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-
prolyl-(L)-tyrosine, tart-butyl ester (100 mg, 0.184 mmole) dissolved in
dry dimethylformamide (1.0 mL) was added anhydrous potassium
carbonate (76.3 mg, 0.552 mmol) and iodomethane {52.3 mg, 0.736
mmole). The reaction mixture was stirred vigorously at 25° C
overnight under a dry nitrogen atmosphere. Ethyl acetate (30 mL)
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was added and the solution acidified with aqueous 5% citric acid to
pH = 5. The layers were separated and the aqueous layer was
extracted with ethyl acetate (3 x 20 mL). Organic layers were
combined and washed successively with water and saturated salt
solution, and dried over anhydrous magnesium sulfate. After
filtration, the solvent was removed by rotoevaporation and the residue
dissolved in a minimum of dichloromethane. This solution was
loaded onto a 1000 micron silica gel Chromatotron plate and purified
by gradient elution with 10-50% ethyl acetate in hexane to afford N-
(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-methyl-tyrosine, tert-
butyl ester as an ofl=white powder (?6 mg, 74% yield).
MS: m/e 557.5 (M+1)'.
400 MHz 1H-NMR (CDC13) 81.44 (s, 9H), 1.56-1.69 (m, 3H), 2.08-2.11
(m, 1H), 2.95 (dd, J = 14.0, 6.68 Hz, 1H), 3.09-3.16 (m, 2H), 3.35-3.40 (m,
1H), 3.75 (s, 3H), 4.07-4.09 (m, 1H), 4.66 (dd, J = 13.8, 6.4 Hz, 1H), 6.78
(d, J = 8.4 Hz, 2H), 7.04 (d, J = 8.6 Hz, 3H), 7.57 (t, J = 1.8 Hz, 1H), 7.70
(d, J = 1.8 Hz, 2H).
Step D: N-(3 5-Dichlorobenzenesulfon lv )-(L)-prolvl-(L) O meth,
tyrosine.
To a solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-
prolyl-(L)-O-methyl-tyrosine, tert-butyl ester (50 mg, 0.090 mmole)
dissolved in dry dichloromethane (0.3 mL) and anisole (5 ~.L) at 0°C
under a dry nitrogen atmosphere was dropwise added a 50% v/v
solution of trifluoroacetic acid in dichloromethane (276uL, 1.8
mmole). After the addition was completed, the ice bath was removed,
and the reaction mixture allowed to stir vigorously for 2.5 hr. The
reaction mixture was treated with dichloromethane (20 mL) and 5%
aqueous sodium bicarbonate to pH = 5. After separation of phases,
the aqueous layer was extracted with dichloromethane (2 x 10 mL).
The organic layers were combined and successively washed with
water and saturated salt solution. The solution was dried over
anhydrous magnesium sulfate and filtered. The solvent was
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removed by rotoevaporation and the residue dissolved in a minimum
of dichloromethane. This solution was loaded onto a 1000 micron
silica gel plate on a Chromatotron eluted with 1-10% methanol in
dichloromethane to afford N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-O-methyl-tyrosine as a light brown powder (28.5 mg, 63% yield).
MS: m/e 501.2 (M+1)+.
400 MHz'H-NMR (CD30D) 81.56-1.65 (m, 2H), 1.74-1.85 (m, 1H), 1.86-
1.88 (m, 1H), 3.01 (dd, J = 13.9, 6.4 Hz, 1H), 3.16-3.24 (m, 2H), 3.3?-3.43
(m, 1H), 3.72 (s, 3H), 4.12 (dd, J = 8.5, 3.4 Hz, 1H), 4.45 (br t, J = 5.7 Hz,
1H), 6.79 (d, J = 8.6 Hz, 2H), 7.15 (d, J = 8.6 Hz, 2H), 7.80 (br m, 3H).
The following compounds were prepared by the
procedures described in Example 299 using the appropriate
alkylating or acylating agent in Step C:
Compound Name MS
(300) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O- 577.4
benzyl-tyrosine


(301)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-543.5


n-butyl-tyrosine


(302)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-526.4


cyanomethyl-tyrosine


(303)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-547.4


(2-methoxyethyl)-tyrosine


(304}N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-559.4


(2-ethoxyethyl)-tyrosine


(305)N-(benzenesulfonyl)-(L}-prolyl-(L)-O-(2-477.0


methoxyethyl)-tyrosine
(306) N-(benzenesulfonyl}-(L)-prolyl-(L)-O-(2- 491.2
ethoxyethyl)-tyrosine
(307) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O- 584.3
( 1-pyrrolidinylcarbonyl)-tyrosine
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(308)N-(benzenesuifonyl)-(L)-prolyl-(L)-O-(1- 516.3


pyrrolidinylcarbonyl)-tyrosine


(309)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-618


(tent-butyl acetate)-tyrosine


(310)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-599.1


(4-morphoiinyl-carbonyl)-tyrosine


(311)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-543.3


( 1-(2-propanonyl)-tyrosine


(312)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-598


prolyl-(L)-O-( 1-pyrrolidinylcarbonyl)-tyrosine


(3I3)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-632.1


prolyl-(L)-O-(tent-butyl acetate)-tyrosine


(314)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-559.3


prolyl-(L)-O-(2-ethoxyethyl)-tyrosine


(315)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-559.4


(acetic acid)-tyrosine, methyl ester
(316)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-545.2


(acetic acid)-tyrosine


(317)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-557.3


prolyl-(L)-O-( 1-(2-propanonyl)-tyrosine


(318)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-612.4


prolyl-(L)-O-( 1-pyrrolidinylcarbonyl
)-tyrosine,


methyl ester


(319)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-614.2


prolyl-(L)-O-(4-morpholinyl-carbonyl)-tyrosine


(320)N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O-580.3


(2-pyrrolylcarbonyl)-tyrosine


(321)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-634.4


prolyl-(L)-O-(N-phenyl-N-methylaminocarbonyl)-


tyrosine


(322)N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-600.3


prolyl-(L)-O-(N,N-diethyl-aminocarbonyl)-


tyrosine
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(323) N-(3-chlorobenzenesulfonyl)-(L)-2(S)-methyl- 580.3
prolyl-(L)-O-(4-morpholinyl-carbonyl )-tyrosine
(324) N-(3,5-dichiorobenzenesulfonyl)-(L)-2(S)-methyl- 628.6
prolyl-(L)-O-(N,N-diisopropyl-aminocarbonyl)-
tyrosine
(325) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O- 591.3
(benzoyl)-tyrosine
(326) N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-(L)-O- 583.3
(cyclopentanoyl)-tyrosine
* m/e: (M + 1 (H+))+ or (M + 18 (NH4+))+
EXAMPLE 327
N-(3 5-Dichlorobenzenesulfon l~p~lvl (L) O (5 tetrazolvl)methvl
~os'
Step A: N-(3.5-dichlorobenzenesulfon lv )-(L)-prQlyl-(L)-O
~vanometh~-tyrosing, tert but 1 ester
To a solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-
prolyl-(L)-tyrosine, tent-butyl ester (200 mg, 0.368 mmole, obtained
from Example 299, Step A) dissolved in 2.0 mL of dry
dimethylformamide was added bromoacetonitrile (353.1 mg, 2.94
mmole) and anhydrous potassium carbonate (152.6 mg, 1.I0 mmole).
The reaction mixture was stirred vigorously under a dry nitrogen
atmosphere at 40°C overnight. The reaction mixture was then
diluted with ethyl acetate and acidified with 5% aqueous citric acid to
pH = 5. After separation of the organic layers, the aqueous layer was
washed with fresh ethyl acetate (3X). The combined organic layers
were successively washed with water, saturated salt solution, and
then dried over anhydrous magnesium sulfate. The residue obtained
after filtration and removal of solvents was purified on a 1000 micron
Chromatotron plate by gradient elution using 10-8-5-4-2-1:1
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Hexane:EtoAc. This afforded 150.4 mg (70% yield) of the title
compound as an off white powder.
MS.: (ESI) m/e 582.4 (M+1)+.
1H-NMR 400 MHz (CDCl3) 81.44 (s, 9H), 1.56-1.69 (m, 3H), 2.08-2.11
(m, 1H), 3.00 (dd, J = 14.0, 6.68 Hz, 1H), 3.05-3.13 (m, 1H), 3.21 (dd, J =
14.0, 6.69 Hz, 1H), 3.35-3.51 (m, 1H), 4.09 (dd, J = 8.5, 3.4 Hz, 1H), 4.68
(dd, J = 13.8, 6.4 Hz, 1H), 4.73 {s, 2H), 6.89 (d, J = 8.7 Hz, 2H), 7.09 (d, J
= 8.6 Hz, 2H), 7.15 (d, J = 8.7 Hz, 2H), 7.58 (distorted m, 1H), 7.70-7.73
(distorted m, 2H).
Step B: N-(3.5-Dichlorobenzenesulfonyl)-(LL,pro~l-(L)-O-
(5-tetrazolvl)meth~3~osine, tert-bu .1 ester
A mixture of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-O-cyanomethyl-tyrosine, tent-butyl ester (82.0 mg, 0.141 mmol)
and f trimethyltin azide { 101.4 mg, 0.493 mmol) in 6 mL of dry toluene
was stirred at reflux for 1 day. The reaction mixture was cooled to
room temperature and concentrated in vacuo. The residue was
treated with 6 mL of dry methanol and 3 g of silica gel and stirred
vigorously overnight at room temperature. This slurry was
concentrated to give a powder. This was vacuum-dried and then
added as a slurry in methylene chloride to a 4.0 x 7.0 cm cartridge of
Flash-40 silica gel and eluted with 10% methanol in methylene
chloride. The fractions containing the desired product were combined
and concentrated to yield 33.0 mg (38.2% yield) of the titled compound
as a white powder.
Mass spectrum (ESI) m/e 630.1 (M+18)+.
1H-NMR 400 MHz (CD30D) b 1.41 (s, 9H), 1.61-1.92 (m, 3H), 2.08-2.11
(m, 1H), 2.97-3.01 (distorted m, 1H), 3.09 (dd, J = 14.0, 6.2 Hz, 1H),
3.24-3.28 (m, 1H), 3.39-3.46 (m, 1H), 4.17-4.21 (m, 1H), 4.52 (dd, J =
14.0, 5.9 Hz, 1H), 5.37 (s, 2H), fi.99 (d, J = 8.7 Hz, 2H}, 7.18 (d, J = 8.7
Hz, 2H), 7.78-7.80 (distorted m, 3H), 8.15 {d, J = 8.1 Hz, 1H).
Step C: N-(3.5-Dichlorobenzenesulfon l~~rolyl-(L)-O-
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{5-tetrazolvl)methy~vr. osir.~g
A mixture of N-(3,5-dichlorobenzenesulfonyl)-(L)-prolyl-
(L)-O-(5-tetrazolyl)methyl-tyrosine, tert-butyl ester (30 mg, 0.0489
mmol) was dissolved in 2 mL of dry methylene chloride and was
cooled in an ice bath. A solution of 1/1 v/v of trifluoroacetic acid (55.7
mg, 0.489 mmol) and methylene was added, which was stirred
vigorously for three hr ice temperature . A stream of dry nitrogen
was applied to remove the solvents and the residue was loaded onto a
reverse phase prep-plate (RP-18wF2sas 0.2 mm 20 x 20 cm, EM
Science) using a minimal amount of methylene chloride and eluted
with 40:60 water/acetonitrile. The product band was collected and
extracted with 10% methanol/methylene chloride, concentrated to
provide 5.0 mg (18% yield) of the titled compound as a white foam
material.
Mass spectrum (ESI) m/e 569.3 (M+1)+.
1H-NMR 500 MHz (CD30D) 81.61-1.87 (m, 3H), 2.05 (distorted m, 1H),
3.02 (dd, J = 14.0, 8.1 Hz, 1H), 3.18 (dd, J = 14.1, 5.2 Hz, 1H),3.23-3.28
(m, 1H), 3.39-3.43 (m, 1H), 4.22 (t, J = 6.0 Hz, 1H), 4.64 (dd, J = 8.0, 5.3
Hz, 1H), 5.41 {s, 2H), 6.99 (distorted d, J = 2.1 Hz, 2H), 7.22 (distorted d,
J = 1.8 Hz, 2H), 7.76-7.78 (m, 3H).
~PLE 328
N-(3 5-Dichlorobenzenesulfonyl) (L) 2(S) methyl rolyl (L) NE benz,~l
h~stidine -
Step A: -t- uty~ycarbonvl-(L)-2lS) methyl p 1'ne
2(S)-Methyl-proline (4.98 g, 38.55 mmol) was dissolved in
dioxane (40 mL) and water (40 mL) to give a suspension. Triethyl
amine (11.4 gm, 46.27 mmol) was added, followed by the addition of 2-
(tert-butoxycarbonyloxyimino)-2-phenylacetonitrile (BOC-ON, ,5.85
gm, 57.83 mmol). The reaction mixture was stirred at room
temperature overnight to give a yellow solution. The reaction was
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quenched with water (150 mL) and diethyl ether (225 mL). The
organic layers were separated and the ether layer extracted with
water (80 mL). The combined aqueous layers were cooled to 0°C and
treated with 2N hydrochloric acid to pH = 2, and then extracted with
ethyl acetate (3 x 150 mL). The combined organic layers were dried
with over anhydrous sodium sulfate, filtered and concentrated to
yield ?.24 g (82% yield) of the titled compound as a white solid (mp =
119-125°C).
Mass spectrum (ESI) m/e 230.1 (M+1)+.
1H-NMR 400 MHz (CD30D) S 1.41 (s, 9H), 1.49 (s, 3H), 1.85-1.99 (m,
3H), 2.13-2.25 (m, 1H), 3.43-3.54 (m, 2H).
Step B: N-t-Butvloxvcarbonyl-(L)-2(S)-methyl-prolyl-(L)-NE-
benzvl-histidine., meth3il ester.
A mixture of N-t-butyloxycarbonyl-(L)-2{S)-methyl-
proline (300 mg, 1.31 mmol) and of (L)-N~-benzyl-histidine, methyl
ester dihydrochloride (339.28 mg, 1.31 mmol) in dry
dimethylformamide (5 mL) and methylene chloride (2.5 mL) was
stirred at room temperature. Diisopropylethyl amine
(684.6 ~,L, 3.93 mmol) was added followed by the addition of
benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphosphate
hexafluorophosphosphate (PyBOP, 681.6 mg, 1.31 mmol) and the
mixture was stirred overnight. This reaction mixture was treated
with 2N hydrochloric acid, water, and ethyl acetate. The layers were
separated and the aqueous layer was extracted with ethyl acetate
(3X). The combined organic layers were washed with saturated
sodium bicarbonate, water, saturated salt solution and dried over
anhydrous magnesium sulfate. After filtration and removal of
solvent by rotoevaporation, the residue was purifed by flash
chromatography on silica gel and eluted with 10-9-8-?-6-5-4-3-2-1:1
Hexane:ethyl acetate and finally with 1-2% methanol/methylene
chloride. The fractions containing the desired material were
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combined and concentrated to yield 357.8 mg {58% yield) of the titled
compound as a sticky white foam.
Mass spectrum (ESI) m/e 4?1.5 (M+1)+.
400 MHz (CD30D) 81.34 (s, 9H), 1.43 (distorted s, 3H), 1.62-2.05 (m,
4H), 2.98-3.11 (m, 2H), 3.38-3.42 (m, 1H), 3.47-3.55 (m, 1H), 3.66 (s, 3H),
4.66-4.70 (m, 1H), 5.16 (distorted s, 2H), 6.95 (s, 1H), 7.26-7.38 (m, 5H),
7.86 (s, 1H), 8.09 (s, 1H).
Step C: (L_)-2(S)-Methyl-pr~olyl-(L)-NE-
benzvl-histidine. methyl ester, dihvdrochloride
A mixture of N-t-butyloxycarbonyl-(L)-2(S)-methyl-prolyl-
(L)-N-benzyl-histidine, methyl ester (272.5 mg, 0.649 mmol) and
hydrochloric acid~g~/ethyl acetate (14.0 mL, 58.4 mmol) in dry ethyl
acetate (2 mL) was stirred at room temperature for one hour.
Methylene chloride was added and solvents were removed by
rotoevaporation. The residue was dried under high vacuum
overnight and gave 235.1 mg (97.6% yield) of the titled compound.
Mass spectrum (CI) m/e 371.3 (M+1)+.
'H-NMR 400 MHz (CDsOD) 8 1.43 (s, 3H), 1.87-1.93 (m, 1H), 2.01-2.13
(m, 2H), 2.32-2.37 (m, 1H), 3.14-3.21 (m, 1H), 3.29-3.38 (m, 4H), 3.71 (s,
3H), 4.77 (dd, J = 10.1, 5.3 Hz, 1H), 5.39 (s, 2H), 7.40-7.43 (m, 5H), 9.05
(distorted s, 1H).
Step D: N-(3.5-Dichlorobenzen sulfonyl)-(L)-2(S)-meth,
prolvl-(L)-NE-benzyl-h,_'stidine, methyl ester
(L)-2(S)-methyl-prolyl-(L)-N-benzyl-histidine, methyl
ester, dihydrochloride (191.3 mg, 0.516 mmol was dissolved in dry
tetrahydrofuran (5 mL) and dry dimethylformamide (2.5 mL).
Diisopropylethyl amine (269.8 ~,L, 1.55 mmol) and 4, 4'-
dimethylaminopyridine were added to this solution. After cooling to
5°C for 5 minutes, a solution of 3,5-dichlorobenzenesulfonyl chloride
(190.2 mg, 0.774 mmol) in dry tetrahydrofuran (2.5 mL) was added to
the reaction mixture which was allowed to reach room temperature
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overnight. This reaction mixture was treated with water and ethyl
acetate. The aqueous layer was extracted with ethyl acetate (3X). The
organic layers were combined and successively washed with water
and saturated salt solution and dried with anhydrous magnesium
sulfate. After filtration, the solvents were removed by
rotoevaporation. The residue was purifed on a 4.0 x 7.0 cm cartridge
of Flash-40 silica gel and eluted 1-2-3-4-5% methanol/methylene
chloride to yield 116.6 mg (39% yield) of the titled compound.
Mass spectrum (CI) m/e 579.1 (M+1}+.
1H-NMR 400 MHz (CDC13) 8 1.67 (s, 3H), 1.72-1.86 (m, 2H), 1.91-1.98
(m, 1H), 2.30-2.35 (m, 1H}, 3.12 (dd, J = 15.0, 4.76 Hz, 1H), 3.18 (dd, J =
14.6, 6.02 Hz, 1H), 3.33-3.39 (m, 1H), 3.66 (s, 3H), 4.7? (dd, J = 6.11, 1.27
Hz, 1H), 5.04 (s, 2H), 6.76 (s, 1H), 7.12-7.15 (m, 2H), 7.29-7.35 (m, 3H),
7.72 (distorted d, J = 1.99 Hz, 2H), 7.99 (distorted s, 2H).
Step E: N-(3,5-Dichlorobenzenesulfonvl)-(L)-2(S)-methyl-
prol,~-(L)-Ne-benzyl-histi dine.
A mixture of N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-
methyl-prolyl-(L)-N-benzyl-histidine, methyl ester ( 115.5 mg, 0.199
mmol) in 0.2N sodium hydroxide in ethanol (1.2 mL) was stirred at
room temperature for 4 hours. The reaction mixture was treated
with ethyl acetate and 5% citric acid to pH = 3-4. The aqueous layer
was extracted with ethyl acetate (3X). The combined organic layers
were washed with saturated salt solution and dried over anhydrous
magnesium sulfate. The solution was filtered and the sovlents were
removed removed by rotoevaporation. The residue was purified on a
4.0 x 7.0 cm cartridge of Flash-40 silica gel eluted with 15%
methanol/methylene chloride to yield 51.2 mg (45.5% yield) of the
titled compound as a light brown foam.
Mass spectrum (ESI) m/e 565.4 (M+1)+.
1H-NMR 400 MHz (CDCD3) 8 1.28 (s, 3H), 1.75-1.84 (m, 3H), 2.10-2.14
(m, 1H}, 3.06-3.12 (m, 1H), 3.24-3.29 (m, 2H), 3.31-3.42 (m, 2H), 4.46-
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4.49 (m, 1H), 5.23 (s, 2H), 7.18 (s, 1H), 7.30-7.37 (m, 5H), 7.74-7.79 (m,
3H), 8.34 (broad s,1H).
N-Benzenesulfonv -(L)- rolvl-2-ammo 2 norbornanecarboxvlic acid
Step A: 2-Amino-2-norbornanecarboxvlic acid, methyl ester
hydrochoride.
To 25 mL of methanol at 0 °C was added thionyl chloride
(2.4 mL, 32 mmol). After stirring at 0 °C for 5 min, 2-amino-2-
norbornanecarboxylic acid (1.0 g, 6.4 mmol) was added in one
portion, and the mixture was heated at reflux for 16 h. The mixture
was concentrated to give the product (1.2 g, 92%) as a white solid.
Step B: N-Benzenesulfonvl-(L)-pro yl-2-amino-2
norbornanecarboxylic acid methyl ester
To a solution of 2-amino-2-norbornanecarboxylate,
methyl ester hydrochloride (400 mg, 2.0 mmol), N-benzenesulfonyl-
(L)-proline (510 mg, 2.0 mmol), 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (306 mg, 2.0 mmol), 1-
hydroxbenzotriazole (202 mg, 2.0 mmol) in 4 mL of tetrahydrofuran at
0 °C was added N-methyl morpholine (0.22 mL, 2.0 mmol). After 15
min at 0 °C, the reaction mixture was stirred at room temperature
for 16 h, and was concentrated in vacuo. The residue was purified by
flash column chromatography on silica gel eluted with 10:1
methylene chloride%thyl acetate to give the title compound (478 mg,
59%) as a mixture of diastereomers.
MS: calculated for C20H26N205S 406; found m/e 417 (M+H+), 423
(M+NH4+).
Step C: N- Benzenesulfonvl-(T )-prolvl-2-amino 2
norbornanecarboxylic acid
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A solution of N-phenylsulfonyl-(L)-prolyl-2-amino-2-
norbornanecarboxylic acid, methyl ester (210 mg, 0.2 mmol) in 3 mL
of 1:1 aqueous sodium hydroxide ( 1 M) and methanol was stirred at
room temperature for 2 weeks. The reaction was quenched with
concentrated hydrochloric acid (0.2 mL), and the resulting mixture
was partitioned between saturated salt solution and ethyl acetate.
The product was extracted with ethyl acetate and was purified by
flash chromatography on silica gel eluted with 100:5:1 methylene
chloride/methanol/acetic acid to give the product as a mixture of
diastereomers.
MS: calculated for C19H24N205S, 392; found m/e 393 (M+H+), 410
(M+NH4+)
EXAMPLE 330
N-Benzenesulfonyl-(L)-prolyl-3(R)-meth ~~1-phenylalanine
Step A: N-Benzenesulfonyl-(L)-prod-3(R)-meth
phenylaianine, methyl ester.
The title compound was prepared by the procedure
described in Example 289 Steps A - C starting from (L)-3(R)-methyl-
phenylalanine (prepared by the procedure of Hruby and coworkers:
Tetrahedron, ~, 4$, 4733).
Step B: N-Benzenesulfon~~rolvl-3(R)-methyl-
Rhenylalanine,
A solution of N-phenylsulfonyl-(L)-prolyl-(L)-3(R)-
methyl-phenylalanine, methyl ester (23 mg, 0.053 mmol) in 1.0 mL of
1:1 tetrahydrofuran/water at 0 °C was added lithium hydroxide
hydrate (12 mg, 0.033 mmol) and hydrogen peroxide (30%, 33 mL,
0.033 mmol). The reaction was allowed to warm up to 18 °C over 2 hr.
The reaction was quenched with dilute sodium thiosulfate and 1 M
hydrochloric acid, and the resulting mixture was partitioned between
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saturated salt solution and ethyl acetate. The product was extracted
with ethyl acetate and purified by flash column chromatography on
silica gel eluted with 50:50:1 ethyl acetate/hexane/acetic acid to 20:1
ethyl acetate/acetic acid to give the product (17 mg, 77%).
MS: calculated for C21H24N2O5S, 416; found m/e 417 (M+H~ ), 434
(M+NH4+)
1H-NMR (500 Mhz, CD30D) 8 8.2-7.2 (lOH, m), 4.65 (1H, d), 4.23 (1H,
dd), 3.48-3.36 (2H, m), 3.23 (1H, m), 2.0-L2 (4H, m), 1.38 (3H, d)
E~~PLE 331
N-Phenvlsulfonvl-(L)-prolvl-(L)-2 3-methano-phenylalanine and N
Phenvlsulfonvl-(L)-prolvl-(D)-2,3-methano-phenvla~lanine
Step A: N-Phenvlsulfonvl-(L)-prol 1-v (L)-~,"3-methano-
nhenvlalanine methyl ester and N Phenvlsulfon l~prol3~l (D) 2 3
met~ano=phen~rlalanine, methyl ester
The title compounds were prepared by the procedure
described in Example 289, Steps A-C starting from E-2,3-
methanophenylalanine, methyl ester hydrochloride (prepared by the
procedure of Stammers and coworkers: J. Org. Chem., 1982, 47,
3270). Under the described conditions, reaction of diazomethane with
Z-2-phenyl-4-benzylidene-5-oxazolinone (Aldrich) gave a 4:1 mixture
of Z-1,5-diphenyl-6-oxa-4-azaspiro(2,4)hept-4-ene-7-one and E-1,5-
diphenyl-6-oxa-4-azaspiro(2,4)hept-4-ene-7-one, and the minor
diastereomer was carried on to E-2,3-methanophenylalanine methyl
ester hydrogen chloride salt as described. Subsequent peptide
coupling (51 mg scale) afforded a 1:1 mixture of diastereomers, which
were partially separated on silica gel eluting with 4:4:1 methylene
chloride/hexane/ethyl acetate.
Top isomer: 'H-NMR (500 Mhz, CD30D) 8 8.0-7.1 (10 H, m), 4.18 (1H,
dd), 3.60 (1H, ddd), 3.30 (3 H, S), 3.4-3.2 (1H, m), 2.96 (1 H, dd), 2.18
(1H, dd), 2.1-1.8 (3H, m), 1.7-1.6 (1H, m), 1.58 (1H, dd)
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Bottom isomer: 1H-NMR (500 Mhz, CD30D) 8 8.0-7.2 (10 H, m), 4.24 {
1H, dd), 3.66 (1 H, ddd), 3.30 (3 H, S), 3.26 (1H, ddd), 2.88 (1 H, dd), 2.22
(1H, dd), 2.1-1.8 (3H, m), 1.66-1.60 (1H, m), 1.53 (1H, dd)
Step B: N-Phenylsulfon 1-~prolvl-(L)-2 3-methano-
phenvlalanine and N-phenylsulfon 1-~prol 1-y (D)-2 3-methano-
phenylalanine.
To a solution of the top isomer of N-phenylsulfonyl-(L)-
prolyl-2,3-methanophenylalanine, methyl ester (15 mg, 0.035 mmol)
in 0.6 mL of 1:1 tetrahydrofuran/water was added lithium hydroxide
hydrate (15 mg, 0.35 mmol), and the mixture was stirred at room
temperature for 15 hr. The reaction was quenched with concentrated
hydrochloric acid (0.2 mL), and the resulting mixture was
partitioned between brine and ethyl acetate. The product was
extracted with ethyl acetate and was purified by flash
chromatography on silica gel eluted with 100:5:1 methylene
chloride/methanol/acetic acid to give the product in quantitative yield.
MS: calculated for C21H22N205S, 414; found m/e 415.3 (M+H+), 432.3
(M+NH4+)
'H-NMR (500 Mhz, CD30D) 8 8.0-7.0 (10 H, m), 4.10 (1H, dd), 3.60 (1H,
ddd), 3.27 (1H, ddd), 2.84 (1 H, dd), 2.18 (1H, dd), 2.1-1.8 (3H, m), 1.66-
1.56 (1H, m), 1.57 (1H, dd).
The bottom isomer was hydrolyzed in the same fashion
as described for the top isomer:
MS: calculated for C21H22N2O5S, 414; found m/e 415.2 (M+H+), 432.2
(M+ NH4+ )
'H-NMR (500 Mhz, CD30D) 8 8.0-7.1 (lOH, m), 4.06 ( 1H, dd), 3.66 (I
H, ddd), 3.27 (1H, ddd), 2.86 (1 H, dd), 2.19 (1H, dd), 2.1-1.8 (3H, m),
1.68-1.58 (1H, m), 1.52 (1H, dd).
EXAMPLE 332
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N-(3 5-Dichlorobenzen sulfowl) (T ~ 2(S) meth~p~glyl (I 4 (5
((1H.3H)-1.3-dimethylR~nidine-2 4-dione))
p ~Pnylalanine
Step A: N-(3-FluorobenzeneRWfo~ ly )-(L)=,prolvl (L) 4
trimeth lstannyl,~nylalanine, tert bu~vl enter
A solution of N-(3-fluorobenzenesulfonyl)-(L)-prolyl-(L)-4-
iodophenylalanine, tert-butyl ester ( 1.0 gm, 1.53 mmol),
hexamethylditin (411 ~,L, 2.14 mmol), triphenylphosphine (8 mg, 0.03
mmol), lithium chloride (71 mg, 1.68 mmol), and
tetrakis(triphenylphosphine)palladium(0) (88 mg, 0.077 mmol) in 1,4-
dioxane (10 mL) was heated to 95°C under a dry nitrogen atmosphere
for 1.5 hr. The solution was cooled and diluted with ethyl acetate ( 100
mL) and successively washed with 1N sodium hydroxide solution
(2X) and saturated salt solution (1X). After drying over anhydrous
magnesium sulfate, the solution was filtered and the solvent removed
by rotoevaporation. The residue was purified by silica gel column
chromatography eluted with 10% acetone in hexanes to yield N-(3-
fluorobenzenesulfonyl )-(L)-prolyl-(L)-4-
(trimethylstannyl)phenylalanine, tert-butyl ester (577 mg, 54% yield).
MS: m/e 658 (M + 18; NH4+).
N-(3,5-Dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-
(L)-4-trimethylstannylphenylalanine, tert-butyl ester was prepared
from N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-methyl-prolyl-(L)-4-
iodophenylalanine, tert-butyl ester by an analogous procedure.
Step B: N-(3.5-Dichlorobenzenesulfonvl)-(L)-2(~1 m~~prolyl
(L)-4-(5-((1H 3H)-1 3-dimeth~pvr~imidine 2 4 dion phenvlalanine~
butyl ester
A solution of N-(3,5-dichlorobenzenesulfonyl)-(L)-2(S)-
methyl-prolyl-(L)-4-trimethylstannylphenylalanine, tent-butyl ester
(?0 mg, 0.1 mmol), (1H,3H)-1,3-dimethyl-5-iodo-pyrimidine-2,4-dione
(40 mg, 0.15 mmol) and tetrakis-triphenylphosphine palladium (4
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mg, 0.003 mmol) in dry dimethylformamide (1 mL) was heated in an
oil bath at 100o C for 1 hr under a dry nitrogen atmosphere. After
cooling, the solvent was removed by rotoevaporation under high
vacuum. The residue was purified by flash column chromatography
on silica gel eluted with 15% acetone in hexanes to give the title
compound as a light yellow solid (27 mg, 40% yield).
MS: (m/e) 696 (M + 18 (NH4+)).
Step C: N-(3.5-Dichlorobenzenesulfonvl)-(L)-2(S)-methyl-prol ~~1-,
(L)-4-(5-((1H.3H)-1.3-dimethvlpyrimidine-2,4-dione))nhenylalanine
The tert-butyl ester of N-(3,5-dichlorobenzenesulfonyl)-
(L)-2(S)-methyl-prolyl-(L)-4-(5-(( 1H,3H)-1,3-dimethylpyrimidine-2,4-
dione))-phenylalanine, tent butyl ester (24 mg, 0.035 mmol) was
stirred in a solution of trifluoroacetic acid (170 ~.L, 2.2 mmol) in
methylene chloride ( 1.0 mL) according to the procedure described in
Example 225, Step E to yield the title compound.
MS: (m/e) 640 (M + 18 (NH4+)).
EXAMPLE 333
Inhibition of VLA-4 Dependent Adhesion to BSA-CS-1 Conjugate
Step A. Preparation of CS-1 Coated Plates
Untreated 96 well polystyrene flat bottom plates were coated
with bovine serum albumin (BSA; 20 ~.g/ml) for 2 hours at room
temperature and washed twice with phosphate buffered saline (PBS).
The albumin coating was next derivatized with 10 ~.g/ml 3-(2-
pyridyldithio) propionic acid N-hydroxysuccinimide ester (SPDP), a
heterobifunctional crosslinker, for 30 minutes at room temperature and
washed twice with PBS. The CS-1 peptide (Cys-Leu-His-Gly-Pro-Glu-Ile-
Leu-Asp-Val-Pro-Ser-Thr), which was synthesized by conventional solid
phase chemistry and purified by reverse phase HPLC, was next added to
the derivatized BSA at a concentration of 2.5 ~,g/ml and allowed to react
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for 2 hours at room temperature. The plates were washed twice with
PBS and stored at 4°C.
Step B. Preparation of Fluorescentl~eled Jurkat Cells
Jurkat cells, clone E6-1, obtained from the American Type
Culture Collection (Rockville, MD; cat # ATCC TIB-152) were grown and
maintained in RPMI-1640 culture medium containing 10% fetal calf
serum (FCS), 50 units/ml penicillin, 50 ~.g/ml streptomycin and 2 mM
glutamine. Fluorescence activated cell sorter analysis with specific
monoclonal antibodies confirmed that the cells expressed both the a4
and ~i1 chains of VLA-4. The cells were centrifuged at 400xg for five
minutes and washed twice with PBS. The cells Were incubated at a
concentration of 2 x 106 cells/ml in PBS containing a 1 ~,M concentration
of a fluorogenic esterase substrate (2', T-bis-(2-carboxyethyl)-5-(and -6)-
carboxyfluorescein, acetoxymethyl ester; BCECF-AM; Molecular Probes
Inc., Eugene, Oregon; catalog #B-1150) for 30-60 minutes at 37°C
in a 5%
C02/air incubator. The fluorescently labeled Jurkat cells were washed
two times in PBS and resuspended in RPMI containing 0.25% BSA at a
final concentration of 2.0 x 106 cells/ml.
StepC. A. say Procedure
Compounds of this invention were prepared in DMSO at
100x the desired final assay concentration. Final concentrations were
selected from a range between 0.001 nM-100 ~.M. Three u,L of diluted
compound, or vehicle alone, were premixed with 300 ~,L of cell
suspension in 96-well polystyrene plates with round bottom wells. 100 ~.L
aliquots of the cell /compound mixture were then transferred in
duplicate to CS-1 coated wells. The cells were next incubated for 30
minutes at room temperature. The non-adherent cells were removed by
two gentle washings with PBS. The remaining adherent cells were
quantitated by reading the plates on a Cytofluor II fluorescence plate
reader (Perseptive Biosystems Inc., Framingham, MA; excitation and
emission filter settings were 485 nm and 530 nm, respectively). Control
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wells containing vehicle alone were used to determine the level of cell
adhesion corresponding to 0% inhibition. Control wells coated with BSA
and crosslinker (no CS-1 peptide) were used to determine the level of cell
adhesion corresponding to 100% inhibition. Cell adhesion to wells coated
with BSA and crosslinker was usually less than 5% of that observed to
CS-1 coated wells in the presence of vehicle. Percent inhibition was then
calculated for each test well and the IC5o was determined from a ten
point titration using a validated four parameter fit algorithm.
EXAMPLE 334
Antagonism of VLA-4 Dependent Binding to VCAM-Ig Fusion Protein
Step A. Preparation of VCAM-Ig.
i5 The signal peptide as well as domains 1 and 2 of human
VCAM (GenBank Accession no. M30257) were amplified by PCR using
the human VCAM cDNA (R & D Systems) as template and the following
primer sequences: 3'-PCR primer:5'-AATTATAATTTGATCAACTTAC
CTGTCAATTCTTTTACAGCCTGCC-3';
5'-PCR primer:
5'-ATAGGAATTCCAGCTGCCACCATGCCTGGGAAGATGGTCG-3'.
The 5'-PCR primer contained EcoRI and PvuII restriction
sites followed by a Kozak consensus sequence (CCACC) proximal to the
initiator methionine ATG. The 3'-PCR primer contained a BcII site and
a splice donor sequence. PCR was performed for 30 cycles using the
following parameters: 1 min. at 94°C, 2 min. at 55°C, and 2 min.
at
72°C. The amplified region encoded the following sequence of human
VCAM-1:
MPGKMVVILGASNILWIMFAASQAFKIETTPESRYLAQIGDSVSLTC
STTGCESPFFSWRTQIDSPLNGKVTNEGTTSTLTMNPVSFGNEHSYLC
TATCESRKLEKGIQVEIYSFPKDPEIHLSGPLEAGKPITVKCSVADVY
PFDRLEIDLLKGDHLMKSQEFLEDADRKSLETKSLEVTFTPVIEDIGKV
LVCR,AKLHIDEMDSVPTVRQAVKEL. The resulting PCR product of
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650 by was digested with EcoRI and BclI and ligated to expression vector
pIg-Tail (R & D Systems, Minneapolis, MN) digested with EcoRI and
BamHI. The pIg-Tail vector contains the genomic fragment which
encodes the hinge region, CH2 and CH3 of human IgGl (GenBank
Accession no. 2173?0). The DNA sequence of the resulting VCAM
fragment was verified using Sequenase (US Biochemical, Cleveland,
OH). The fragment encoding the entire VCAM-Ig fusion was
subsequently excised from pIg-Tail with EcoRI and NotI and ligated to
pCI-neo (Promega, Madison, WI) digested with EcoRI and NotI. The
resulting vector, designated pCI-neo/VCAM-Ig was transfected into
CHO-Kl (ATCC CCL 61) cells using calcium-phosphate DNA
precipitation (Specialty Media, Lavalette, NJ). Stable VCAM-Ig
producing clones were selected according to standard protocols using
0.2-0.8 mg/mI active 6418 (Gibco, Grand Island, N~, expanded, and cell
supernatants were screened for their ability to mediate Jurkat adhesion
to wells previously coated with 1.5 ~,g/ml (total protein) goat anti-human
IgG (Sigma, St. Louis, MO). A positive CHO-Kl/~7CAM-Ig clone was
subsequently adapted to CHO-SFM serum-free media (Gibco) and
maintained under selection for stable expression of VCAM-Ig. VCAM-
Ig was purified from crude culture supernatants by affinity
chromatography on Protein A/G Sepharose {Pierce, Rockford, IL)
according to the manufacturer's instructions and desalted into 50 mM
sodium phosphate buffer, pH 7.6, by ultrafiltration on a YM-30
membrane (Amicon, Beverly, MA).
Step B. Preparation of ~I-VCAM-Ig
VCAM-Ig was labeled to a specific radioactivity greater that
1000 Ci/mmole with 1251-Bolton Hunter reagent (New England Nuclear,
Boston, MA; cat # NEX120-0142) according to the manufacturer's
instructions.The labeled protein was separated from unincorporated
isotope by means of a calibrated HPLC gel filtration column (G2000SW;
?.5 x 600 mm; Tosoh, Japan) using uv and radiometric detection.
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Step C. V AM-Ig Binding Assav.
Compounds of this invention were prepared in DMSO at
100x the desired final assay concentration. Final concentrations were
selected from a range between 0.001 nM-100 ~.M. Jurkat cells were
centrifuged at 400xg for five minutes and resuspended in binding buffer
(25 mM HEPES, 150 mM NaCl, 3 mM KCl, 2 mM glucose, 0.1% bovine
serum albumin, pH 7.4). The cells were centrifuged again and
resuspended in binding buffer supplemented with MnCl2 at a final
concentration of 1 mM. Compounds were assayed in Millipore MHVB
multiscreen plates (cat# MHVBN4550, Millipore Corp., MA) by making
the following additions to duplicate wells: (i) 200 ~.L of binding buffer
containing 1 mM MnCl2; (ii) 20 ~,L of 1251-VCAM-Ig in binding buffer
containing 1 mM MnCl2 (final assay concentration ~ 100 pM); (iii) 2.5 ~.L
of compound solution or DMSO; (iv) and 0.5 x 106 cells in a volume of 30
~.L. The plates were incubated at room temperature for 30 minutes,
filtered on a vacuum box, and washed on the same apparatus by the
addition of 100 ~.L of binding buffer containing 1 mM MnCl2. After
insertion of the multiscreen plates into adapter plates (Packard,
Meriden, CT, cat# 6005178), 100 ~.L of Microscint-20 (Packard cat#
6013621) was added to each well. The plates were then sealed, placed on
a shaker for 30 seconds, and counted on a Topcount microplate
scintillation counter (Packard). Control wells containing DMSO alone
were used to determine the level of VCAM-Ig binding corresponding to
0% inhibition. Contol wells in which cells were omitted were used to
determine the level of binding corresponding to 100% inhibition. Binding
of 1251-VCAM-Ig in the absence of cells was usually less than 5% of that
observed using cells in the presence of vehicle. Percent inhibition was
then calculated for each test well and the IC6o was determined from a
ten point titration using a validated four parameter fit algorithm.
EXAMPLE 335
Antagonism of a,I3z Dependent Binding to VCAM-Ig Fusion Protein
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Step A. g~ZCel1 ling
RPMI-8866 cells (a human B cell line a4+~il'(37'"; a gift from
Prof. John Wilkins, University of Manitoba, Canada) were grown in
RPMI/10% fetal calf serum/ 100 U penicillin/100 p,g streptomycin/2 mM
L-glutamine at 37°C, 5 % carbon dioxide. The cells were pelleted at
1000
rpm for 5 minutes and then washed twice and resuspended in binding
buffer (25 mM Hepes, 150 mM NaCI , 0.1 % BSA, 3 mM KCI, 2 mM
Glucose, pH 7.4).
Step B. VCAM-Ig Binding Assay
Compounds of this invention were prepared in DMSO at
100x the desired final assay concentration. Final concentrations were
selected from a range between 0.001 nM-100 p,M. Compounds were
assayed in Millipore MHVB multiscreen plates (Cat# MHVBN4550) by
making the following sequential additions to duplicate wells: (i) 100
~.1/well of binding buffer containing 1.5 mM MnCl2; (ii) 10 pl/well lzsl-
VCAM-Ig in binding buffer (final assay concentration < 500 pM); (iii) 1.5
~1/well test compound or DMSO alone; (iv) 38 ~,1/well RPMI-8866 cell
suspension (1.25 x 106 cells/well). The plates were incubated at room
temperature for 45 minutes on a plate shaker at 200 rpm, filtered on a
vacuum box, and washed on the same apparatus by the addition of 100
p.L of binding buffer containing 1 mM MnCl2. After insertion of the
multiscreen plates into adapter plates (Packard, Meriden, CT, cat#
6005178), 100 ~.L of Microscint-20 (Packard cat# 6013621) was added to
each well. The plates were then sealed, placed on a shaker for 30
seconds, and counted on a Topcount microplate scintillation counter
(Packard). Control wells containing DMSO alone were used to
determine the level of VCAM-Ig binding corresponding to 0% inhibition.
Wells in which cells were omitted were used to determine the level of
binding corresponding to 100% inhibition. Percent inhibition was then
calculated for each test well and the ICSO was determined from a ten
point titration using a validated four parameter fit algorithm.
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