Note: Descriptions are shown in the official language in which they were submitted.
~ =
W o 96/10034 2 2 0 13 4 8 PCTrUS95/12224
THIOL-FREE INHIBITORS OF FARNESYL-PROTEIN TRANSFERASE
RELATED APPLICATIONS
The present patent application is a continll~tion-in-part
application of copending application Serial No. 08/314,974, filed
September29, 1994.
BACKGROUND OF THE INVENTION
The Ras protein is part of a sign~lling pathway that links cell
surface growth factor receptors to nuclear signals initi~ing cellular
proliferation. Biological and biochemical studies of Ras action indicate
that Ras functions like a G-regulatory protein. In the inactive state, Ras
is bound to GDP. Upon growth factor receptor activation Ras is induced
to exchange GDP for GTP and undergoes a conformational change. The
GTP-bound form of Ras propagates the growth stimulatory signal until
the signal is termin~t~d by the intrinsic GTPase activity of Ras, which
returns the protein to its inactive GDP bound form (D.R. Lowy and D.M.
Willumsen, Ann. Rev. Biochem. 62:851-891 (1993)). Mutated ras genes
are found in many hllm~n cancers, including colorectal carcinoma,
exocrine pancreatic carcinoma, and myeloid leukemias. The protein
products of these genes are defective in their GTPase activity and
constitutively tr~n~mit a growth stimulatory signal.
Ras must be localized to the plasma membrane for both
normal and oncogenic functions. At least 3 post-translational
modifications are involved with Ras membrane localization, and all 3
modifications occur at the C-terminus of Ras. The Ras C-terminus
contains a sequence motif termed a "CAAX" or "Cys-Aaal-Aaa2-Xaa"
box (Cys is cysteine, Aaa is an aliphatic amino acid, the Xaa is any amino
acid) (Willumsenetal.,Nature310:583-586(1984)). Dependingonthe
specific sequence, this motif serves as a signal sequence for the enzymes
farnesyl-protein transferase or geranylgeranyl-protein transferase, which
catalyze the alkylation of the cysteine residue of the CAAX motif with a
C1s or C20 isoprenoid, respectively. (S. Clarke., Ann. Rev. Biochem.
WO 96/10034 == = ~ PCTIUS95/12224
22,~3~
61:355-386 (1992); W.R. Schafer and J. Rine, Ann. Rev. Genetics
30:209-237 (1992)). The Ras protein is one of several proteins lhat are
known to undergo post-translational farnesylation. Other farnesylated
~ro~i,ls include the Ras-related GTP-binding proteins such as Rho,
5 fungal mating factors, the nuclear l~min.~, and the gamma subunit of
transducin. James, et al., J. Biol. Chem. 269, 14182 (1994) have
identified a peroxisome associated protein Pxf which is also far~esylated.
James, et al., have also suggested that there are farnesylated proteins of
unknown structure and function in addition to those listed above.
Inhibition of farnesyl-protein transferase has been shown to
block the growth of Ras-transformed cells in soft agar and to modify
other aspects of their transformed phenotype. It has also been
demonstrated that certain inhibitors of farnesyl-protein transferase
selectively block the processing of the Ras oncoprotein intracellularly
(N.E. Kohl et al.~ Science, 260: 1934- 1937 (1993) and G.L. James et al.,
Science, 260:1937-1942 (1993). Recently, it has been shown that an
inhibitor of farnesyl-protein transfera$e blocks the growth of ras-
dependent tumors in nude mice (N.E. Kohl et al., Proc. Natl. Acad. Sci
U.S~., 91:9141-9145 (1994) and induces regression of m~mm~ry and
20 salivary carcinomas in ras transgenic mice (N.E. Kohl et al., Nature
Medicine, 1:792-797 (1995).
It has recently been shown that farnesyl-protein transferase
inhibitors are inhibitors of proliferation of vascular smooth muscle cells
and are therefore useful in the prevention and thereapy of arteriosclerosis
25 and diabetic disturbance of blood vessels (JP H7-112930).
Indirect inhibition of farnesyl-protein transferase i~l vivo has
been demonstrated with lovastatin (Merck & Co., Rahway, NJ) and
compactin (Hancock et al., ibid; Casey et al., ibid; Schafer et al., Science
245:379 (1989)). These drugs inhibit HMG-CoA reductase, the rate
30 limi~ing enzyme for the production of polyisoprenoids including farnesyl
pyrophosphate. Farnesyl-protein transferase utilizes farnesyl
pyrophosphate to covalently modify the Cys thiol group of the Ras
CAAX box with a farnesyl group (Reiss et al., Cell, 62:81-88 (1990);
Schaber et al., J. Biol. Chem., 265:14701-14704 (1990); Schafer et al.,
-- =
WO 96/10034 PCr/US95112224
220~3~8 .' - .
Science, 249:1133-1139 (19gO); Manne et al., Proc. Natl. Acad. Sci USA,
87:7541-7545 (1990)). Inhibition of farnesyl pyrophosphate biosynthesis
by inhibiting HMG-CoA reductase blocks Ras membrane loc~li7~tion in
cultured cells. However, direct inhibition of farnesyl-protein transferase
would be more specific and attended by fewer side effects than would
occur with the required dose of a general inhibitor of isoprene
biosynthesis.
Inhibitors of farnesyl-protein transferase (FPTase) have been
described in two general classes. The first are analogs of farnesyl
diphosphate (FPP), while the second class of inhibitors is related to the
protein substrates (e.g., Ras) for the enzyme. The peptide derived
inhibitors that have been described are generally cysteine cont~ining
molecules that are related to the CAAX motif that is the signal for protein
prenylation. (Schaber et al., ibid; Reiss et. al., ibid; Reiss et al., PNAS,
88:732-736 (1991)). Such inhibitors may inhibit protein prenylation
while serving as alternate substrates for the farnesyl-protein transferase
enzyme, or may be purely competitive inhibitors (U.S. Patent 5,141,~51,
University of Texas; N.E. Kohl et al., Science, 260:1934-1937 (1993);
Graham, et al., J. Med. Chem., 37, 725 (1994)). In general, deletion of the
thiol from a CAAX derivative has been shown to dramatically reduce the
inhibitory potency of the compound. However, the thiol group
potentially places limitations on the therapeutic application of FPTase
inhibitors with respect to ph~rm~cokinetics, pharmacodynamics and
toxicity. Therefore, a functional replacement for the thiol is desirable.
It is, therefore, an object of this invention to develop
tetrapeptide-based compounds which do not have a thiol moiety, and
which will inhibit farnesyl transferase and the post-translational
function~ tion of the oncogene Ras protein. It is a fur~er object of
this invention to develop chemotherapeutic compositions cont~inin~
the compounds of this invention and methods for producing the
compounds of this invention.
WO g6/10034 PCTIUS95/12224
2?~0~34~,
SUMMARY OF THE INVENTION
The present invention comprises analogs of the CAAX
motif of the protein Ras that is modified by farnesylation in vivo.
These CAAX analogs inhibit the farnesylation of Ras. Furthermore,
5 these CAAX analogues differ from those previously described as
inhibitors of Ras farnesyl transferase in that they do not have a thiol
moiety. The lack of the thiol offers unique advantages in terms of
improved ph~rm~cokinetic behavior in ~nim~lc, prevention of thiol-
dependent chemical reactions, such as rapid autoxidation and
disulfide formation with endogenous thiols, and reduced systemic
toxicity. Further contained in this invention are chemotherapeutic
compositions co~ illg these farnesyl transferase inhibitors and
methods for their production.
WO 96/10034 2 2 0 1 3 '~ 8 i PCT/US95112224
S _
The compounds of this invention are illustrated by the
form~ e:
( IR )r ~9 ~ R~R2b z R ~R5b
V - A (CR 2)nA2(CR12)n - W- (CR12)p N X~ ~R1
R3 R4
(R8)r ~9 Z R~a R2b Z R5a R5b
V ~ A1(CR12)nA2(CR12)n ~ W ~ (CR12)p NR12 X ~R1~0R
R3 R4
1511
HOCH2(,CH2)q
V - A1(CR12)nA2(CR12)n - W - (CR12)J~N1~2~x~ ~N~OH
R3 R4
111
and
(R8)r R9 Z R2a R2b z ~q
V - A1(CR12)nA2(CR12)n - W - (CR 2)p NR12 X ~R1
` R3 R4
IV
WO 96/10034 PCT/US95112224
22~13~8 - ~
DETAILED DESCRIPTTON OF THE ~VENTION
The compounds of this invention inhibit the A
farnesylation of Ras. In a first embodiment of this invention, the Ras
farnesyl transferase inhibitors are illustrated by the formula I:
(R8)r 1~9 ~ R~R2b Z R5a R5b
V - A1(CR12)nA2(CR12)n - W - (CR 2)p R12 X ~R14
wherein:
Rl is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
R10O-, Rl lS(O)m-~ Rl0C(O)NRl0-~ CN, NO2,
(Rl0)2N-C(NRl0)-, Rl0C(O)-, Rl0OC(O)-, N3,
-N(Rl0)2, or Rl lOC(O)NRl0-,
c) C l -C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R100-,
Rl lS(O)m, Rl0C(O)NRl0-~ CN, (R10)2N-C(NR10)-
Rl0C(O)-, Rl0OC(O)-, N3, -N(Rl0~2, or
Rl lOC(O)NR10;
R2a and R2b are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl -C20 alkyl, C2-c2o
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02, R100-, Rl lS(O)m-, RlOC(O)NR10- CN
WO 96/10034 PCTIUS95/12224
,~ 2201348
(RlO)2N C(NR10), RlOC(O)-~ RlOOC(O)-~ N3,
-N(R10)2, RllOC(O)NR10- and Cl-C20 alkyl, and
d) Cl-C6 alkyl substituted wit~ an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
R2a and R2b are combined to form - (CH2)s -;
R3 and R4 are independently selected from:
o a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-c2o alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
R3 and R4 are combined to form - (CH2)S -;
R5a and R5b are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-C20 alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocycle group,
WO 96/10034 PCT/IJS95/12224
22Q134~ - 8 -
wherein the substituent is selected from F, Cl, Br,
N(R10)2, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN (R10)2N C(NR10), RlOC(O)-~ RlOOC(O)-, N3,
N(R10)2, Rl loC(o)NRlo-~ -S02N(R10)2,
R 1 1 SO2NR10- and C1 -C20 alkyl, and
d) Cl-C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloaL~cyl; or
o RSa and R5b are combined to form - (CH2)S - wherein one of the carbon
atoms is optionally replaced by a moiety selected from: O, S(O)m,
-NC(O)-, and-N(COR10)-; or
R5a or RSb are combined with R 14 to form a ring such that
R5a ~R5b ~ ~
`N~ is ~H2)t;
R14 R15
-
WO 96/10034 PCTiUS9~/12224
. 22013 18 ` -~ ~
g
X-Y is
~7a
a)~sss~N~
O
~7b
b) \sSS~ N~ss
C)~ 0~5S
( 1)m
d) ~s~S~5
e) _55s~ , or
f) -CH2-CH2-;
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloaLkyl, and
e) C 1 -C6 alkyl substituted with hydrogen or an
; unsubstituted or substituted group selected from aryl, heterocyclic and cycloalkyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
WO 96110034 PCI`/IJS95112224
~2013 ;~
- 10-
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloaLkyl,
e) C 1 -C6 aLkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from. aryl,
heterocyclic and cycloalkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
perf~uoroalkyl, F, Cl, Br, R100-, Rl lS(O)m-,
RlOC(O)NR10-, CN, N02, R102N-C(NR10)-,
R1OC(O)-, R1OOC(O)-, N3, -N(R10)2, or
R1 loc(o)NRlo-~ and
c) Cl-C6 alkyl unsubstituted or substituted by aryl~
heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroalkyl, F, Cl, Br, RlOO, Rl 1
RlOC(O)NH-, CN, H2N-C(NH)-, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or R1 1OC(O)NH-;
R9 is selected from:
a) hydrogen, .,
b) alkenyl, aLkynyl, perfluoroalkyl, F, Cl, Br, R100-,
R1 1S(O)m-, R1OC(O)NR10-, CN, NO2, (R10)2N-
WO 96/10034 PCT/US9~/12224
~ 2201348 . - -
t i
- i1 -
C(NR10)-, RlOC(oj-, RlOOC(03-, N3, -N(R10)2, or
Rl lOC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by
perfluoroalkyl, F, Cl, Br, Rloo-~ Rl lS(o)m
R10C(o)NR10, CN, (R10)2N-C(NR10)-, RlOC(O)-,
R 10oC(O)-, N3, -N(R 1)2, or Rl lOC(O)NR10-;
R10 is independently selected from hydrogen, Cl-C6 alkyl and aryl;
o Rl 1 is independently selected from C1-C6 alkyl and aryl;
R12 is independently selected from hydrogen and C1-C6 alkyl;
R14 is independently selected from hydrogen,C1-C6 alkyl and
benzyl;
R15 is independently selected from hydrogen and C1-C6 alkyl;
Al and A2 are independently selected from: a bond, -CH=CH-,
2 o -C_C-, O, -N(R 10) , -C(O)-, -C(O)NR 10 , -NR 1 C(O)-,
-S(0)2N(R10)-, -N(R10)S(0)2- or S(O)m;
V is selected from:
a) hydrogen,
b) heterocycle,
c) aryl,
d) C1-C20 alkyl wherein from O to 4 non-termin~l
carbon atoms are replaced with a heteroatom selected
from 0, S, and N, and
e) C2-C20 alkenyl;
provided that V is not hydrogen if Al is S(O)m and V is not
hydrogen if A 1 is a bond, n is O and A2 is S(O)m or a bond;
W is a heterocycle;
~L
WO 96/10034 PCIIUS95/12224
2201348 i - --
- 12-
Z is independently H2 or 0,
misO, 1 or2;
nis 0,1,2,30r4,
pis 0,1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
s is 4 or 5; and
t is 3, 4 or 5;
or the pharmaceutically acceptable salts thereof.
In a second embodiment of this invention the prodrugs
of compounds of formula I are illustrated by the formula II:
(R8~ R9 Z R2ap2b Z R5a R5b
V - A (CR 2)nA (CR12)n -W - (CRl2)p N X~ ~R o
R3 R4
Il
wherein:
R1 is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
R100-, Rl 1S(O)m-~ R1OC(O)NR10-~ CN, N02,
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, or Rl 1OC(O)NR10-,
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, R100-,
R1 1S(O)m-, R1OC(o)NRlo-~ CN, (R10)2N C(NR10),
RlOC(O)-, RlOOC(O)-,N3,-N(R10)2,or
Rl lOC(O)NR10
R2a and R2b are independently selected from:
a) a side chain of a naturally occurring amino acid,
WO 96/10034 , , PCT/US95/12224
~ . -; ,. . . .
22al~s~
- 13-
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methioI~ine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl -C20 alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2 RlOO-, Rl lS(O)m-, RlOC(O)NR10-, CN,
10)2N C(NR10), Rloc(o)-~ RlOoc(o)-~ N3,
o -N(R10)2, Rl lOC(O)NR 10 and Cl-C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
R2a and R2b are combined to form - (CH2)s -;
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl -c20 alkyl, C2-c20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R10)2, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
3 0 substituted group selected from aryl, heterocycle and C3-
~,, Clo cycloalkyl; or
R3 and R4 are combined to form - (CH2)s -;
WO 96/10034 i~ r PCT/US95/12224
2201~48
- 14-
RSa and R5b are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
,:2 5 i)methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-c2o alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocycle group,
wherein the substituent is selected from F, Cl, Br,
o N(R10)2, N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
-N(Rl0)2, RllOC(O)NR10-, -SO2N(Rl0)7"
R1 lS02NR10-and C1 -C20 alkyl, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
R5a and R5b are combined to form - (CH2)S - wherein one of the carbon
atoms is optionally replaced by a moiety selected from: O, S(O)m,
20 -NC(O)-, and -N(COR 10)-; or
R5a or R5b are combined with R 14 to form a ring such that
2 5 R ~R ~ ~
~ss`N~ is (~ ((3H2)t;
R14 >~.R15
R6 is
a) substituted or unsubstituted Cl -C8 alkyl, wherein the
substituent on the alkyl is selected from:
1) aryl,
WO 96/10034 PCT/US95/12224
220~8
r ~
- 15-
2) heterocycle,
3) -N(Rl 1)2,
4) -OR10, or
b)
R12 o
1 J~ 13 ~,
X-Y is
~7a
a)~sss~N~s
O
R7b
b)\sss~N~ss
c) ~ O~5s
()m
d) ~5sS~S~5s
2s e)55s~5ss
H
fl -CH2-CH2-;
" R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
WO 96/10034 PCI/US95112224
= - 8-
- 16-
d) unsubstituted or substituted cycloalkyl, and
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsuL~sli~uLed or substituted heterocyclic,
o d) unsubstituted or substituted cycloalkyl,
e) C l-C6 aLkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroaLkyl, F, Cl, Br, R100-, Rl 1 S(O)m,
RlOC(O)NR10-, CN, N02, R102N-C(NR10)-,
RlOC(O)-, R100C(O)-,N3,-N(R10)2,or
R1 loc(o)NRlo-~ and
c) C 1 -C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroalkyl, F, Cl, Br, RlOO-, R1 1S(O)m-,
WO 96/10034 PCT/US95/12224
.
2201318
RlOC(O)NH-, CN, H2N-C(NH)-, RlOC(O)-,
R1OOC(O)-, N3, -N(R10)2, or Rl 1OC(O)NH-;
7 R9 is selected from:
a) hydrogen,
b) alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R100-,
Rl lS(O)m-, RlOC(O)NR10-, CN, NO2, (R10)2N-
C(NR10)- RlOC(O)-~ RlOOC(O)-~ N3, -N(R 1)2, or
R1 loc(o)NRlo-~ and
o c) Cl-C6 alkyl unsubstituted or substituted by
perfluoroalkyl, F, Cl, Br, R100-, Rl lS(O)m-,
R1OC(O)NR10-, CN, (R10)2N-C(NR10)-, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or Rl 1OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
R12 is independently selected from hydrogen and C1-C6 alkyl;
R13 is independently selected from Cl-C6 alkyl;
R14 is independently selected from hydrogen,Cl-C6 alkyl and
benzyl;
R15 is independently selected from hydrogen and Cl-C6 alkyl;
Al and A2 are independently selected from: a bond, -CH=CH-,
-C_C-, -C(O)-, -C(O)NR10-, O, -N(R10)-,
-NRlOC(O)-, -S(O)2N(R10)-, -N(R10)S(0)2- or S(O)m;
V is selected from:
a) hydrogen,
b) heterocycle,
WO 96/10034 PCT/ILJS95/12224
. .
i.
~o ~3 ~
d) Cl-C20 alkyl wherein from O to 4 non-termin~l
carbon atoms are replaced with a heteroatom selecl:ed
from 0, S, and N, and
e) C2-C20 alkenyl;
provided that V is not hydrogen if A 1 is S(O)m and V is not
hydrogen if A 1 is a bond, n is O and A2 is S(O)m or a bond;
W is a heterocycle;
Z is independently H2 or 0;
misO, 1 or2;
nis 0,1,2,30r4;
piS 0,1,2,30r4;
r is O to 5, provided that r is O when V is hydrogen;
s is 4 or S; and
t is 3, 4 or 5;
or the pharmaceutically acceptable salts thereof.
In a third embodiment of this invention, the inhibitors of
farnesyl transferase are illustrated by the formula m:
HocH2(cl~2jq
V - A1(CR12) A2(CR1 ) - W - (CR12)J~N~x~y~N~foH
wherein:
R1 is independently selected from:
WO 96/10034 PCT/US95/12224
~ 22013~8
- 1 9 - ~
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
RlOO, Rl lS(O)m, RlOC(O)NR10-, CN, NO2,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, or R1 lOC(O)NR10-,
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, RlOO-,
Rl 1S(O)m-, RlOC(O)NR10-, CN, (R10)2N-C(NR10)-,
RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
Rl lOC(O)NR10;
R2a and R2b are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl -C20 alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, RlOO-, Rl lS(O)m-, RlOC(O)NR10-, CN,
10)2N C(NR10), Rloc(o)-~ R 10oc(o), N3,
N(RlO)2~ RllOC(O)NR10- and Cl-C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloaL~yl; or
R2a and R2b are combined to form - (CH2)s -;
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
WO 96/10034 PCT/US95/12224
3 ~ 20-
ii) methionine sulfone, and
c) substituted or unsubstituted C1-C20 alkyl, C2-C20
alkenyl, C3-C1o cycloalkyl, aryl orheterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N(R1)2~ NO2, RlO-, RllS(O)m-, RlOC(O)NR10-,
CN, (R10)2N-C(NR10)-, R 10C(o)-, R1OOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C20 allkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle arld C3-
o Clo cycloalkyl; or
R3 and R4 are combined to form - (CH2)s -;
X-Y is
1 5 ~7a
a) ~sss~N~ss
o
R7b
b) \Sss~ N~ss
c) ~ 0~
( 1)m
d) ~5sS~S~
e) ~sss~ , or
H
f) -CH2-CH2-;
WO 96110034 PCT/US95112224
~ 2201~1~
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
e) C1-C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl,
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and C1-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloaLkyl and C 1 -C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroalkyl, F, Cl, Br, R10O-, Rl lS(O)m-,
RlOC(O)NR10-, CN, N02, R102N-C(NR10)-,
WO 96/10034 PCI/US95112224
--
RlOC(O)-, RlOOC(O)-,N3,-N(R10)2,or
R1 lOC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroalkyl, F, Cl, Br, RlOO-, Rl lS(O)m-,
RlOC(O)NH-, CN, H2N-C(NH)-, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or Rl 1OC(O)NH-;
R9 is selected from:
o a) hydrogen,
b) alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R100-,
R11S(O)m-, R1OC(O)NR10-, CN, NO2, (R10)2N-
C(NR10) RlOC(o)-~ Rlooc(o)-~ N3~ -N(Rlo)2~ or
R1 loc(o)NRlo-~ and
c) Cl-C6 alkyl unsubstituted or substituted by
perfluoroalkyl, F, Cl, Br, R100-, Rl lS(O)m-,
RlOC(O)NR10-, CN, (R10)2N-C(NR10)-, RlOC(O)-,
RlOOC(O)-, N3, -N(R10)2, or Rl lOC(O)NR10-;
20 R10 is independently selected from hydrogen, Cl-C6 alkyl and aryl;
R l 1 is independently selected from C1 -C6 alkyl and aryl;
R12 iS independently selected from hydrogen and Cl-C6 aLkyl;
R14 is independently selected from hydrogen,Cl-C6 alkyl and
benzyl;
Al and A2 are independently selected from: a bond, -CH=CH-,
3 o -C_C-, -C(O)-, -C(O)NR 10, O, -N(R 1 0) , -NR 1 C(O)-,
-S(O)2N(R10)-, -N(R1O)S(O)2- or S(O)m,
V is selected from:
a) hydrogen,
WO 96/10034 PCT/US95/12224
22~1348 - !
,.
- 23 -
b) heterocycle,
c) aryl,
d) Cl-C20 alkyl wherein from O to 4 non-terminal
carbon atoms are replaced with a heteroatom selected
from 0, S, and N, and
e) C2-C20 alkenyl;
provided that V is not hydrogen if A l is S(O)m and V is not
hydrogen if A l is a bond, n is O and A2 is S(O)m or a bond;
W is a heterocycle;
Z is independently H2 or 0;
misO, l or2;
n is 0, 1, 2, 3 or 4;
p is 0, l, 2, 3 or4;
q is 0, l or 2;
r is O to 5, provided that r is O when V is hydrogen; and
s is 4 or 5;
or the pharmaceutically acceptable salts thereof.
In a fourth embodiment of this invention the prodrugs of
compounds of formula III are illustrated by the formula IV:
25 (R8) ~9 Z R2a R2b z l q
V - A1(CR12)nA2(CR12)n - W - (CR12)J~N~ix~ ~ R1
R3 R4
IV
wherem:
Rl is independently selected from:
WO 96/10034 PCT/IJS95/12224
3~
- 24 -
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, alkynyl,
RlOO Rl 1S(o)m, R10c(o)NRlo-~ CN, NO2,
(R10)2N C(NR10), R1OC(O)-, R1OOC(O)-~ N3,
-N(R10)2, or R1 1OC(O)NR10-,
c) C 1 -C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl, RlOO-,
Rl lS(O)m-, RlOC(O)NR10-, CN, (R10)2N-C(NR10)-,
RlOC(O)-, RlOOC(O)-, N3, -N(R10)2, or
o Rl lOC(o)NR10;
R2a and R2b are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-C20 alkyl, C2-C20
alkenyl, C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, Rl 1S(O)m-, R1OC(O)NR10-, CN,
(RlO)2N C(NR10), R1OC(O)-, RlOOC(O)-~ N3,
-N(R10)2, Rl lOC(O)NR10- and Cl-C2o alkyl, and
d) C1-C~ alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; or
R2a and R2b are combined to form - (CH2)s -;
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
WO 96/10034 PCTtUS95/12224
~ 2201348
ii) methionine sulfone, and
c) substituted or unsubstituted Cl -C20 alkyl, C2-C20
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
- wherein the substituent is selected from F, Cl, Br,
N(RlO)2~ N02, R100-, Rl lS(O)m-, RlOC(O)NR10-,
CN (R 10)2N C(NR10), R 10c(O)-~ R l OOC(O)-, N3,
N(R10)2~ Rl lOC(O)NR10- and C1-C20 alkyl, and
d) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
ClO cycloalkyl; or
R3 and R4 are combined to form - (CH2)s -;
X-Y is
~7a
a) ~sss~N~ss
o
7b
b) \5sS~ N~ss
c) ~ 0~
( I)m
d) ~ssS~S~
H
e) --SSS~ or
H
f) -CH2-CH2-;
WO 96/10034 PCT/US95/12224
2~013~
- 26 -
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl,
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and C1-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaL~yl;
R8 is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, alkenyl, aLkynyl,
perfluoroalkyl, F, Cl, Br, R100-, Rl 1 S(O)m,
RlOC(O)NR10-, CN, N02, R102N-C(NR10)-,
WO 96/10034 PCTIUS95/12224
22nl3~
R1OC(O)-, R 10oC(O)-, N3, -N(R 1)2, or
R1 1OC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, alkynyl,
perfluoroalkyl, F, Cl, Br, R100-, Rl lS(O)m-,
RlOC(O)NH-, CN, H2N-C(NH)-, RlOC(O)-,
R1OOC(O)-, N3, -N(R10)2, or Rl 1OC(O)NH-;
R9 is selected from:
o a) hydrogen,
b) alkenyl, alkynyl, perfluoroalkyl, F, Cl, Br, R100-,
Rl1S(O)m-, R1OC(O)NR10-, CN, NO2, (R10)2N-
C(NR10)-, RlOC(O)-, R1OOC(O)-, N3, -N(R10)2, or
R1 1OC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by
perfluoroaLkyl, F, Cl, Br, RlOO, Rl lS(o)m
R1OC(O)NR10-, CN, (R10)2N-C(NR10)-, R1OC(O)-,
R1OOC(O)-, N3, -N(R10)2, or R1 1OC(O)NR10-;
20 R10 is independently selected from hydrogen, Cl-C6 alkyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
R12 is independently selected from hydrogen and Cl-C6 alkyl;
R14 is independently selected from hydrogen,Cl-C6 alkyl and
benzyl;
A1 and A2 are independently selected from: a bond, -CH=CH-,
3 0 -C-C-, -C(O)-, -C(O)NR 10, O, -N(R 10) -NR 1 C(O)
-S(O)2N(R10)-, -N(R10)s(o)2- or S(O)m;
V is selected from:
a) hydrogen,
WO 96/10034 PCT/US95/12224
22nl~
- 28 -
b) heterocycle,
c) aryl,
d) Cl-C20 alkyl wherein from O to 4 non-terminal
carbon atoms are replaced with a hetero~tom selected
from 0, S, and N, and
e) C2-C20 alkenyl;
provided that V is not hydrogen if A1 is S(O)m and V is not
hydrogen if Al is a bond, n is O and A2 is S(O)m or a bond;
o W is a heterocycle;
Z is independently H2 or 0;
misO, 1 or2;
n is 0, 1, 2, 3 or 4;
pis 0,1,2,30r4;
q is 0, 1 or2;
r is O to 5, provided that r is O when V is hydrogen; and
s is 4 or 5;
or the pharmaceutically acceptable salts thereof.
In a more preferred embodiment of this invention, the
Ras farnesyl transferase inhibitors are illustrated by the formula I:
V-A1(CR12)nA2(CR12)n-W-(CR12)p ~ R `~OH
wherein:
R1 is independently selected from~
a) hydrogen,
WO 96/10034 PCT/US95/12224
2201348t ..
- 29 -
b) aryl, heterocyclic, cycloalkyl, R100-, -N(R10)2 or
alkenyl,
c) Cl-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, R100-, or -N(R10)2;
R2a is selected from:
a) a side chain of a naturally occurring amino acid,
wherein the amino acid is selected from ~l~nine,
leucine, isoleucine and valine;
o b) substituted or unsubstituted Cl-clo alkyl, C2-clo
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02 R10O-, Rl lS(o)m, RlOc(o)NRlo-~ CN,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
-N(R10)2, Rl 1 OC(O)NR10- and C1 -C20 alkyl, and
c) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl; and
R2b iS selected from hydrogen and Cl-C6 alkyl; or
R2a and R2b are combined to form - (CH2)s -,
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-Clo alkyl, C2-Clo
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02, R100-, R1 1S(O)m-, RlOC(O)NR10-, CN,
WO 96/10034 PCTIUS95/12224
- , ............. . .
3 ~S
- 30 -
(R10)2N-C(NR10)-, RlOC(O)-, R1OOC(O), N3,
-N(R10)2, Rl 1OC(O)NR10- and C1-C20 alkyl, and
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl;
R5a is selected from:
a) a side chain of a naturally occurring amino acid,
wherein the amino acid is selected from
o methionine and gl~lt~mine,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone, and
c) substituted or unsubstituted Cl-clo alkyl, C2-C10
aL~enyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, RlOO-, Rl lS(O)m-, RlOC(O)NR10-, CN,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, N3,
N(R10)2, Rlloc(o)NRlo-~ -SO2N(R10)2,
R1 1SO2NR10- and C1-C20 alkyl, and
d) C 1 -C6 alkyl substituted with an unsubstituted or
su~stituted group selected from aryl, heterocycle and C3-
Clo cycloaLkyl;
RSb is selected from:
a) hydrogen, and
b) C1-C3 alkyl; or
3 o R5a or R5b are combined with R 14 to form a ring such that
WO 96/10034 PCIIUS95/12224
22013~8
R ~R ~_~
~`N~ is (~ (~H2)t;
R14 ,~R15
X-Y is
R7a
a) ~S5s~N~sss
R7b
b) \5sS~ N~ss
c)
H
d) ~ , or
H
e) -CH2-CH2-;
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
3 0 c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLkyl;
WO 96110034 PCT/US95/12224
2~20~348 - 32 -
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 7-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
o d) unsubstituted or substituted cycloalkyl,
e) Cl-C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and C 1 -C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLlcyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaL~yl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R8 is selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-c6 alkenyl, C2-c6 alkynyl, Cl-C6
perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN,
WO 96/10034 PCTIIJS95/12224
2 2 0 1 3 4 8
- 33 -
N02, (R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-,
-N(R10)2, or Rl 1OC(O)NR10-, and
c) Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl,
R100-, RlOc(o)NRlo-~ (R10)2N-C(NR10)-,
s RlOC(O)-, RlOOC(O)-, -N(R10)2, orRllOC(O)NR10-;
R9 is selected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl,
o F, Cl, RlOO-, R1 1S(O)m-, R1OC(O)NR10-, CN, NO2,
(Rlo)2N-c(NRlo)-~ Rloc(o)-~ RlOoC(o)- -N(R10)2
or R1 lOC(O)NR10-, and
c) Cl-C6 alkyl unsubstituted or substituted by C1-C6
perfluoroalkyl, F, Cl, RlOO-, Rl lS(o)m
RlOC(O)NR10-, CN, (R10)2N-C(NR10)-, RlOC(O)-,
RlOOC(O)-, -N(R10)2, or Rl 1OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl and aryl;
20 Rl 1 is independently selected from Cl-C6 alkyl and aryl;
R12 is independently selected from hydrogen and Cl-C6 alkyl;
R14 is independently selected from hydrogen and C1-C6 alkyl;
R15 is independently selected from hydrogen and C1-C6 alkyl;
A1 and A2 are independently selected from: a bond, -CH=CH-,
- -C_C-, -C(O)-, -C(O)NR 10 , O, -N(R 1 0) , -NR 1 C(O)-,
-S(O)2N(R10)-, -N(R1O)S(O)2- or S(O)m;
V is selected from:
WO 96/10034 PCTIUS95/12224
r~
- 34 -
a) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl,
indolyl, quinolinyl, isoquinolinyl, and thienyl,
b) aryl,
c) Cl-C20 alkyl wherein from O to 4 non-termin~l
carbon atoms are replaced with a heteroatom selected
from 0, S, and N, and
d) C2-C20 alkenyl;
provided that V is not hydrogen if Al is S(O)m and V is not
hydrogen if A1 is a bond, n is O and A2 is S(O)m or a bond;
W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, piperidinyl, 2-oxopiperidinyl, indolyl,
quinolinyl, or isoquinolinyl;
Z is independently H2 or 0;
misO, 1 or2,
20 nisO,1,2,30r4;
pis 0,1,2,30r4;
r is O to 2;
s is 4 or 5; and
t is 3, 4 or 5;
2s
or ~e pharmaceutically acceptable salts thereof.
In a second more preferred embodiment of this
invention, the prodrugs of the preferred compounds of formula I are
30 illustrated by the formula II:
WO 96/10034 PCT/US95/12224
2201348 i~
- 35 -
V A1(CR12)nA2(CR12) W (CR1 )/I~N~X~Y~N~OR6
R3 R
11
wherein:
R1 is independently selected from:
a) hydrogen,
o b) aryl, heterocyclic, cycloalkyl, R l OO, -N(R 1 )2 or
alkenyl,
c) C 1 -C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, RlOO-, or -N(R10)2;
R2a is selected from:
a) a side chain of a naturally occurring amino acid,
wherein the amino acid is selected from alanine,
leucine, isoleucine and valine;
b) substituted or unsubstituted Cl -Clo alkyl, C2-C10
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, RlOO-, Rl 1S(O)m-, RlOC(O)NR10-, CN,
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, Rl lOC(O)NR10- and C1 -C20 alkyl, and
c) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; and
- R2b is selected from hydrogen and Cl-C6 alkyl; or
r R2a and R2b are combined to form - (CH2)s -;
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
WO 96/10034 PCT/US9~/12224
220~3 ~ '
- 36 -
.
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-clo alkyl, C2-clo
alkenyl, C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, R1 1S(O)m-, R1OC(O)NR10-, CN,
10)2N C(NR10), Rloc(o)-~ R10oc(o)-~ N3,
o -N(R1O)æ R1 1OC(O)NR10- and C1-C20 alkyl, and
d) Cl-C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl;
R5a is selected from:
a) a side chain of a naturally occurring amino acid,
wherein the amino acid is selected from
methionine and glllt~mine,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone, and
c) substituted or unsubstituted Cl-clo alkyl, C2-clo
aLkenyl, C3-C1o cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
NO2, R1OO-, R1 1S(O)m-, R1OC(O)NR10-, CN,
(R10)2N C(NR10), R10C(O)-, R1OOC(O)-~ N3,
N(R10)2, Rl loC(o)NRlo-~ -S02N(R10)2,
R1 lS02NR10- and Cl-C20 alkyl, and
3 0 d) Cl -C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl;
R5a is selected from:
wo 96/10034 PCT/USg~112224
~ 2201398
- 37 -
a) hydrogen, and
b) C1-C3 alkyl, or
RSa or R5b are combined with R 14 to form a ring such that
R ~R
~NR14~S~ iS (~
R6 is
a) substituted or unsubstituted Cl-Cg alkyl, wherein the
substituent on the alkyl is selected from:
1 ) aryl,
2) heterocycle,
3) -N(Rl 1)2,
4) -OR10, or
b)
R12 0
~,10~'R13
r
WO 96/10034 PCT/US95/12224
220i3a~ -38 -
X-Y is
,R
a) ~sss~N~
O
R7b
b) \sSs~N~sss
c) ~,,O~
d) ~SSs~ , or
e) -CH2-CH2-;
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLkyl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R7b is selected from
a) hydrogen,
-
WO 96110034 PCT/US95/12224
2 2 ~ 1 3 ~ 8
- 39 -
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl,
e) C1-C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
o an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and C 1 -C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaLkyl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R8 is selected from:
a) hydrogen,
b) C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6
perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN,
N2, (Rlo)2N-c(NRlo)-~ Rloc(o)-~ RlOoC(o~
-N(R10)2, or R1 1OC(O)NR10-, and c) C1-C6 alkyl
substituted by C1-C6 perfluoroalkyl, R1OO-,
RlOC(O)NR10-~ (RlO)2N-c(NRlo)-~ RlOC(O)-,
RlOOC(O)-, -N(R10)2, or Rl lOC(O)NR10-;
R9 is selected from:
a) hydrogen,
WO 96/10034 PCT/US95/12224
22013 l~ - ~
- 40 -
b) C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 perfluoroalkyl,
F, Cl, R10O-, Rl 1S(O)m-, R10C(O)NR10-, CN, NO2,
(R10)2N-C(NR10)-, RlOC(O)-, RlOOC(O)-, -N(R10)2,
or R1 1OC(O)NR10-, and
c) C1-C6 alkyl unsubstituted or substituted by Cl-C6
perfluoroalkyl, F, Cl, R10O-~ R1 1S(o)m
RlOC(O)NR10-, CN, (R10)2N-C(NR10)-, RlOC(O)-,
R10OC(O)-, -N(R10)2, or R1 1OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl and aryl,
R11 is independently selected from C1-C6 alkyl and aryl;
R12 is independently selected from hydrogen and C1-C6 alkyl;
R13 is 1 ,1-dimethylethyl;
R14 is independently selected from hydrogen and Cl-C6 alkyl;
R15 is independently selected from hydrogen and Cl-C6 alkyl;
A 1 and A2 are independently selected from: a bond, -CH=CH-,
-C-C-, -C(O)-, -C(O)NR10-, O, -N(R 10) -NR10C(O)
-S(O)2N(R10)-, -N(R10)S(0)2- or S(O)m;
V is selected from:
a) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl,
indolyl, quinolinyl, isoquinolinyl, and thienyl,
b) aryl,
c) Cl-C20 alkyl wherein from 0 to 4 non-terrnin~l
carbon atoms are replaced with a heteroatom selected
from O, S, and N, and
d) C2-C20 alkenyl;
WO 96/10034 PCTIUS95112224
2~1 3q 8 - - -
- 41 -
provided that V is not hydrogen if A 1 is S(O)m and V is not
hydrogen if A 1 is a bond, n is O and A2 is S(O)m or a bond;
5 W iS a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, piperidinyl, 2-oxopiperidinyl, indolyl,
quinolinyl, or isoquinolinyl;
Z is independently H2 or 0;
misO, l or2;
nisO,1,2,30r4;
pis 0,1,2,30r4;
r is O to 2;
15 sis40r5;and
t is 3, 4 or 5;
or the pharmaceutically acceptable salts thereof.
In a third more preferred embodiment of this invention,
the inhibitors of farnesyl transferase are illustrated by the formula III:
HOCH2(CH2)q
V - A1 (CR1 2)nA2(CR 1 2)n - W - (cR12) J~N ~ N ~OH
111
30 wherein:
R1 is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, R100-, -N(R 1)2 or
alkenyl,
WO 96110034 PCTIUS95/12224
~20134~
- 42 -
c) C 1 -C6 aLkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, R100-, or-N(R10)2;
R2a is selected from: ~
a) a side chain of a naturally occurring amino acid,
wherein the amino acid is selected from alanine,
leucine, isoleucine and valine;
b) substituted or unsubstituted Cl -C10 alkyl, C2-Clo
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
o wherein the substituent is selected from F, Cl, Br,
N02, R100-, Rl lS(O)m-, R1OC(O)NR10-, CN,
(R10)2N C(NR10), RlOc(o)-~ RlOOC(O)-, N3,
-N(R 1)2, Rl lOC(O)NR10- and Cl-C20 alkyl, and
c) C 1 -C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
C1o cycloalkyl; and
R2b is selected from hydrogen and C1-C6 alkyl; or
R2a and R2b are combined to form - (CH2)S -;
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
occurring amino acid which is:
i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted C1-Clo alkyl, C2-Clo
alkenyl, C3-Clo cycloalkyl, aryl or heterocyclic group,
3 o wherein the substituent is selected from F, Cl, Br,
N02, R100-, R1 lS(O)m-, RlOC(O)NR10- CN
(R10)2N-C(NR1o), Rloc(o)-~ RlOoC(o)~ N3,
-N(R10)2, R1 1OC(O)NR10- and Cl-C20 alkyl, and
WO 96/10034 PCItUS9~112224
.
22ol3~8
- 43 -
d) Cl-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloaLkyl;
5 X-Y is
R7a
a) ~S5s~Nsss
R7b
b) \sss~N~Ss
c) ~SsS~O~ss
d) ~SSs~ , or
H
e) -CH2-CH2-;
R7a is selected from
. a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
e) Cl-C6 aLkyl substituted with hydrogen or an
3 unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
WO 96/10034 PCT/IJS95/12224
22013~8
- 44 -
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R7b is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl,
e) Cl-C6 alkyl substituted with hydrogen or an
o unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaL~yl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloaL~yl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloaL~yl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
2s and thienyl;
R8 is selected from:
a) hydrogen,
b) Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6
perfluoroalkyl, F, Cl, R100-, RlOC(O)NR10-, CN,
N2, (Rlo)2N-c(NRlo)-~ RlOc(o)-~ RlO
-N(R10)2, or Rl lOC(O)NR10-, and
WO 96/10034 PCT/US95112224
2~2013~ 8
- 45 -
c) Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl,
R100-, RlOC(O)NR10-~ (Rlo)2N-c(NRlo)-~ RlOC(O)-
R1OOC(O)-, -N(R10)2, or Rl 1OC(O)NR10-;
R9 is selected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluoroalkyl,
F, Cl, R1OO-, R1 1S(O)m-, R1OC(O)NR10-, CN, NO2,
(Rlo)2N-c(NRlo)-~ RlOc(o)-~ RlOoC(o)- -N(R10)2
o or Rl 1OC(O)NR10-, and
c) C1-C6 alkyl unsubstituted or substituted by Cl-C6
per~luoroaL~yl, F, Cl, RlOO-, Rl lS(o)m
R10C(o)NR10, CN, (R10)2N-C(NR10)-, R1OC(O)-,
R1OOC(O)-, -N(R10)2, or R1 1OC(O)NR10-;
R10 is independently selected from hydrogen, C1-C6 alkyl and aryl;
R11 is independently selected from C1-C6 alkyl and aryl;
R12 is independently selected from hydrogen and C1-C6 alkyl;
R14 is independently selected from hydrogen and C1-C6 alkyl;
A1 and A2 are independently selected from: a bond, -CH=CH-,
2 5 -C-C-, -C(O)-, -C(O)NR 10 , O, -N(R 1 0) -NR 1 C(O)
-S(O)2N(R10)-, -N(R1O)S(O)2- or S(O)m;
V is selected from:
a) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl,
indolyl, quinolinyl, isoquinolinyl, and thienyl,
b) aryl,
WO 96/10034 PCI/US95/12224
22013~8 46
c) Cl-C20 alkyl wherein from O to 4 non-terrnin~l
carbon atoms are replaced with a heteroatom selected
from 0, S, and N, and
d) C2-C20 alkenyl;
provided that V is not hydrogen if Al is S(O)m and V is not
hydrogen if Al is a bond, n is O and A2 is S(O)m or a bond;
W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, piperidinyl, 2-oxopiperidinyl, indolyl,
quinolinyl, or isoquinolinyl;
Z is independently H2 or 0;
15 m is 0, 1 or 2;
nis 0,1,2,30r4;
pis 0,1,2,30r4;
qisO, 1 or2;
risOto2; and
20 SiS4OrS;
or the pharmaceutically acceptable salts thereof.
In a four~ more preferred embodiment of this invention,
25 the prodrugs of the preferred compounds of formula III are illustrated
by the formula IV:
V A1(CR12)nA2(CR12)n-W-(CR12)J~N1~2~X' ~N~J
IV
wherein:
WO 96/10034 PCT/US95/12224
2201348
- 47 -
Rl is independently selected from:
a) hydrogen,
b) aryl, heterocyclic, cycloalkyl, R100-, -N(R10)2 or
alkenyl,
c) C1-C6 alkyl unsubstituted or substituted by aryl,
heterocyclic, cycloalkyl, alkenyl, R100-, or-N(R10)2;
R2a is selected from:
a) a side chain of a naturally occurring amino acid,
o wherein the amino acid is selected from ~l~nine,
leucine, isoleucine and valine;
b) substituted or unsubstituted Cl -C10 alkyl, C2-C10
alkenyl, C3-C10 cycloalkyl, aryl or heterocyclic group,
wherein the substituent is selected from F, Cl, Br,
N02, R100-, Rl 1S(O)m-, R1OC(O)NR10-, CN,
(R10)2N-C(NR10)-, R1OC(O)-, R1OOC(O)-, N3,
-N(R10)2, R11OC(O)NR10- and C1-C20 alkyl, and
c) C1-C6 alkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl; and
R2b is selected from hydrogen and C1-C6 alkyl; or
R2a and R2b are combined to form - (CH2)s -;
R3 and R4 are independently selected from:
a) a side chain of a naturally occurring amino acid,
b) an oxidized form of a side chain of a naturally
- occurring amino acid which is:
3 o i) methionine sulfoxide, or
ii) methionine sulfone,
c) substituted or unsubstituted Cl-clo alkyl, C2-C10
alkenyl, C3-C1o cycloalkyl, aryl or heterocyclic group,
WO 96/10034 PCTIUS95/12224
~2Q131~
- 48 -
wherein the substituent is selected from F, Cl, Br,
NO2 RlOO, Rl lS(o)m, RlOC(O)NR10-~ CN,
(R10)2N C(NR10), Rloc(o)-7 RlOoc(o)-~ N3,
-N(R10)2, R1 lOC(O)NR10- and C1-C20 alkyl, and
d) Cl-C6 aLkyl substituted with an unsubstituted or
substituted group selected from aryl, heterocycle and C3-
Clo cycloalkyl,
X-Y is
R7a
a) ~Sss~Ns5s~
o
7b
R
b) \~s~,N555
c) ~,O~
H
d) ~5SS~ , or
e) -CH2-CH2-;
R7a is selected from
a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl, and
WO 96/10034 2 2 0 I 3 4 8 PCT/US95/12224
- 49 -
e) Cl-C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R7b is selected from
o a) hydrogen,
b) unsubstituted or substituted aryl,
c) unsubstituted or substituted heterocyclic,
d) unsubstituted or substituted cycloalkyl,
e) C 1 -C6 alkyl substituted with hydrogen or an
unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl,
f) a carbonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloalkyl, and
g) a sulfonyl group which is bonded to an unsubstituted
or substituted group selected from aryl, heterocyclic,
cycloalkyl and Cl-C6 alkyl substituted with hydrogen or
an unsubstituted or substituted group selected from aryl,
heterocyclic and cycloaL~yl;
wherein heterocycle is selected from pyrrolidinyl,
imidazolyl, pyridinyl, thiazolyl, pyridonyl, 2-
oxopiperidinyl, indolyl, quinolinyl, isoquinolinyl,
and thienyl;
R8 is selected from:
a) hydrogen,
WO 96/10034 . PCT/US95/12224
~ ,
22~13 ~8
- so -
b) Cl-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6
per~uoroaLkyl, F, Cl, R100-, RlOC(O)NRlO-, CN,
N2, (Rlo)2N-c(NRlo)-~ Rloc(o)-~ RlO
-N(Rl0)2, or Rl lOC(O)NR10-, and
c) Cl-C6 alkyl substituted by Cl-C6 perfluoroalkyl,
R100-, RlOC(O)NR10-~ (R10)2N-C(NR10)-,
RlOC(O)-, R1OOC(O)-, -N(Rl0)2, or Rl 1 OC(O)NR10-;
R9 is selected from:
a) hydrogen,
b) C2-C6 alkenyl, C2-C6 alkynyl, Cl-C6 perfluor~alkyl,
F, Cl, RlOO-, Rl lS(O)m-, RlOC(O)NRlO-, CN, NO2,
(Rlo~2N-c(NRlo)-~ Rloc(o)-~ RlOoC(o)-~ -N(R10)2,
or R l l OC(O)NRl -, and
c) Cl-C6 alkyl unsubstituted or substituted by Cl-C6
perfluoroaIkyl, F, Cl, RlOO-, Rl lS(o)m
RlOC(O)NRlO-~ CN, (R10)2N-C(NR10)-, RlOC(C~
RlOOC(O)-, -N~RlO)2~ or Rl lOC(O~NR10-;
20 RlO is independently selected from hydrogen, Cl-C6 alkyl and aryl;
R1 l is independently selected from Cl-C6 alkyl and aryl;
Rl2 is independently selected from hydrogen and Cl-C6 alkyl;
R14 is independently selected from hydrogen and Cl-C6 alkyl;
Al and A2 are independently selected from: a bond, -CH=CH-,
-C_C-, -C(O)-, -C(O)NR10-~ 0, N(R10), -NR 10c
-S(O)2N(Rl0)-, -N(RlO)S(O)2- or S(O)m;
V is selected from:
WO 96/10034 PCT/US95112224
22Ql~
- 51 -
a) heterocycle selected from pyrrolidinyl, imidazolyl,
pyridinyl, thiazolyl, pyridonyl, 2-oxopiperidinyl,
indolyl, ~uinolinyl, isoquinolinyl, and thienyl,
b) aryl,
c) Cl-C20 alkyl wherein from O to 4 non-~e""i"~l
carbon atoms are replaced with a heteroatom selected
from 0, S, and N, and
d) C2-C20 alkenyl;
provided that V is not hydrogen if A1 is S(O)m and V is not
o hydrogen if Al is a bond, n is O and A2 is S(O)m or a bond;
W is a heterocycle selected from pyrrolidinyl, imidazolyl, pyridinyl,
thiazolyl, pyridonyl, piperidinyl, 2-oxopiperidinyl, indolyl,
quinolinyl, or isoquinolinyl;
Z is independently H2 or 0;
m is 0, 1 or2;
20 nis 0,1,2,30r4;
pis 0,1,2,30r4;
qisO, 1 or2;
r is O to 2; and
s is 4 or 5;
or the pharmaceutically acceptable salts thereof.
The preferred compounds of this invention are as
follows:
N-[2(S)-(1 -(Phenylmethyl)-lH-imidazol-4-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
WO 96/10034 PCI/US95/12224
-
22013~8
- 52 -
N-[2(S)-(1 -(Phenylmethyl)-lH-imidazol-4-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(Phenylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
5 methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-( 1 -(Phenylmethyl)- 1 H-imidazol-5-ylacetyl)-amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-( 1 -(4-Nitrophenylmethyl)- 1 H-imidazol-4-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
N-[(2S)-( 1 -(4-Nitrophenylmethyl)- 1 H-imidazol-4-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(4-Nitrophenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-( 1 -(4-Nitrophenyl-methyl~- 1 H-imidazol-5 -ylacetyl)amino-3 (S)-
20 methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
25 N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(1 -Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-(1-(1 -Naphthylmethyl)- lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methyl ester
WO 96/10034 PCTIUS95112224
22~I3~
- 53 -
N-[2(S)-( 1 -Farnesyl- 1 H-imidazol-5-ylacetyl)amino-3 (S)-methylpentyl] -
N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-(l -Farnesyl-lH-imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-
5 N-l-naphthylmethyl-glycyl-methionine methylester
N-[2(S)-(1 -Geranyl-lH-imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-
N- 1 -naphthylmethyl-glycyl-methionine
o N-[2(s)-(l-Geranyl-lH-imidazol-5-ylacetyl)amino-3(s)-methylpentyl]
N-l-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(4-Pyridylmethyl)-lH-imidazol-4-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine
N-[2(S)-(1 -(4-Pyridylmethyl)-lH-imidazol-4-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(4-Pyridylmethyl)-lH-imidazol-5-ylacetyl)amino-(3S)-
2 o methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-(1 -(4-Pyridylmethyl)- lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
25 N-[2(S)-(1-(4-Cyanophenylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-(1-(4-Cyanophenylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methylester
N-[2(S)-( 1 -(4-Methoxyphenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-
3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
WO 96/10034 PCTIUS95/12224
- 54 -
N-[2(S)-(1 -(4-Methoxyphenylmethyl)-1 H-imidazol-5-ylacetyl)amino-
3(S)-methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(4-Quinolinylmethyl)- lH-imidazol-5-ylacetyl)amino-3(S)-
5 methylpentyl]-N-1-naphthylmethyl-glycyl-methionine
N-[2(S)-(1 -(4-Quinolinylmethyl)- lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
o N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl] -N- 1 -phenylmethyl-glycyl-methionine
N-[2(S)-(1 -(2-Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-phenylmethyl-glycyl-methionine methyl ester
N-[2(S)-(1 -(2-Naphthylmethyl)- lH-imidazol-5-ylethyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine
N-[2(S)-(1 -(2-Naphthylmethyl)- 1 H-imidazol-5-ylethyl)amino-3(S)-
20 methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
2(S)-[N-2(S)-(1 -(2-Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methyl]pentyloxy-3-phenylpropionyl-methionine sulfone methyl ester
2 5 2(S)-[N-2(S)-(1 -(2-Naphthylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-
methyl]pentyloxy-3-phenylpropionyl-methionine sulfone
2(S)-[N-2(S)-(1 -(2-Naphthylmethyl)- lH-imidazol-5-ylethyl)amino-3(S)-
methyl]pentyloxy-3-phenylpropionyl-methionine methyl ester
2(S)-[N-2(S)-(1 -(2-Naphthylmethyl)- lH-imidazol-5-ylethyl)amino-3(S)-
methyl]pentyloxy-3 -phenylpropionyl-methionine
WO g6110034 PCT/US95/12224
22013~8
.. . ~ .
- 55 -
N-~2(S)-(1 -Methyl-lH-imidazol-4-ylacetyl)-amino-3(S)-methylpentyl]-
N-(1-naphthylmethyl)-glycyl-methionine methyl ester
N-[2(S)-(l -Methyl-lH-imidazol-4-ylacetyl)-amino -3(S)-methylpentyl]-
5 N-(1-naphthylmethyl)-glycyl-methionine
N-[2(S)-N-(2-Naphthylmethyl)- 1 H-imidazol-5-ylacetyl]amino-(3S)-
methylpentyl]-N-(cyclopropylmethyl)-glycylmethionine methyl ester
o N-[(2S)-N-(2-Naphthylmethyl)-1 H-imidazol-5-ylacetyl]amino-(3S)-
methylpentyl] -N-(cyclopropylmethyl) -glycylmethionine
N-[2(S)-[(5(R,S)-Methylpyroglutamyl)amino] -3(S)-methylpentyl]-N-( 1-
naphthylmethyl)-glycylmethionine methyl ester
N- [2(S)- [(5(R,S)-Methylpyroglutamyl)amino] -3 (S)-methylpentyl] -N-( 1-
naphthylmethyl) -glycylmethionine
N-[2(S)-((N-Methylpyroglutamyl)amino)-3(S)-methylpentyl]-N-(l -
2 o naphthylmethyl)-glycyl-methionine
N-[2(S)-((N-Methylpyroglutamyl)-amino)-3(S)-methylpentyl] -N-( 1-
naphthylmethyl)-glycyl-methionine methyl ester
25 N-[2(S)-(N-Formylprolylamino)-3(S)-methylpentyl]-N-(l-
naphthylmethyl)-glycyl-methionine methyl ester
N-[2(S)-(N-Formylprolylamino)-3(S)-methylpentyl]-N-(1 -
naphthylmethyl) -glycyl -methionine
N-[2(S)-(N' -(4-Nitrobenzyl)pyroglutamyl)-amino)-3(S)-methylpentyl]-
N-(l-naphthylmethyl)-glycyl-methionine methyl ester
WO 96/10034 , PCIIIJS95112224
~2ol3~8
- 56 -
N-[2(S)-(N' -(4-Nitrobenzyl)pyroglutamyl)-amino)-3(S)-methylpentyl]-
N-( 1 -naphthylmethyl)-glycyl-methionine
N-[2(S)-((N'-Benzylpyroglllt~m,yl)amino)-3(S)-methylpentyl]-N-(l -
5 naphthylmethyl)-glycyl-methionine methyl ester
N-[2(S)-(N' -Benzylpyroglutamyl)amino)-3(S)-methylpentyl]-N-( 1-
naphthylmethyl) -glycyl -methionine
o N-[2(S)- 1 -(4-Fluorophenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3 (S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
N-[2(S)-1 -(4-Fluorophenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl] -N- 1 -naphthylmethyl-glycyl-methionine
N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(~S)-
methylpentyl]-N-(1-naphthylmethyl)glycyl-methionine isopropyl ester
N-[2(S)-([1 -(4-cyanobenzyl)-lH-imidazol-S-yl]acetylamino)-3(S)-
20 methylpentyl]-N-(l-naphthylmethyl)glycyl-methionine sulfone methyl
ester
N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-(l-naphthylmethyl)glycyl-methionine sulfone
N-[2(S)-([1-(4-cyanobenzyl)-lH-imicl~7.ol-S-yl]acetylarnino)-3(S)-
methylpentyl]-N-( 1 -naphthylmethyl)glycyl-(3-acetylamino)~l~nine
methyl ester
30 N-[2(S~-([1-(4-cyanobenzyl)-lH-imidazol-S-yl]acetylamino)-3(.S)-
methylpentyl]-N-(1 -naphthylmethyl)glycyl-(3-acetylamino)~l~nine
WO 96/10034 PCT/US95/12224
2Z01~48
N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-(1-naphthylmethyl)glycyl-2(RS) amino-3-(2
thienyl)propionic acid methyl ester
5 N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-( 1 -naphthylmethyl)glycyl-2(RS)-amino-3-(2
thienyl)propionic acid
N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
o methylpentyl]-N-( 1 -naphthylmethyl)glycyl-2(S) amino-4-sulfamyl-
butanoic acid methyl ester
N-[2(S)-([ 1 -(4-cyanobenzyl)-1 H-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-(l-naphthylmethyl)glycyl-2(S) amino-4-sulfamyl-
butanoic acid
N- [2(S)-([ 1 -(4-cyanobenzyl)- 1 H-imidazol-5-yl] acetylamino)-3 (S)-
methylpentyl]-N-(l-naphthylmethyl)glycyl-N-methyl methionine methyl
ester
N-[2(S)-([ 1 -(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-(1-naphthylmethyl)glycyl-N-methyl methionine
N-[2(S)-([1 -(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
2 5 methylpentyl]-N-( 1 -naphthylmethyl)glycyl-homoserine lactone
N-[2(S)-([1 -(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-( 1 -naphthylmethyl)glycyl-homoserine
30 N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-( 1 -naphthylmethyl)glycyl-proline methyl ester
N-[2(S)-([ 1 -(4-cyanobenzyl)-1 H-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-( 1 -naphthylmethyl)glycyl-proline
WO 96/10034 PCI/~JS95/12224
220134~ - 58 -
N-[2(S)-([ 1 -(4-cyanobenzyl)-1 H-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-(l-naphthylmethyl)glycyl-D-proline me~yl ester
5 N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(S)-
methylpentyl]-N-( 1 -naphthylmethyl)glycyl-D-proline
N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetylamino)-3(~
methylpentyl]-N-(l-naphthylmethyl)glycyl-L- pipecolinic acid
N-[2(S)-([ 1 -(4-carbomethoxybenzyl)- 1 H-imidazol-5-yl]acetylamino)-
3(S)-methylpentyl]-N-(l-naphthylmethyl)glycyl-methionine methyl ester
N-[2(S)-([ 1 -(4-carbomethoxybenzyl)-1 H-imidazol-5-yl]acetylannino)-
3(S)-methylpentyl]-N-(l-naphthylmethyl)glycyl-methionine
1 -(2-naphthylmethyl)- l H-imidazol-5-ylacetyl-isoleucinyl-
phenyl~l~ninyl-methionine methyl ester
2 o 1 -(2-naphthylmethyl)- 1 H-imidazol-5-ylacetyl-isoleucinyl-
phenyl~l~ninyl-methionine
or the pharmaceutically acceptable salts thereof.
2s Representative compounds of the invention are:
N-[2(S)-(1 -(4-Nitrophenylmethyl)- lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-methionine
WO 96/10034 PCTIUS95/12224
22013~8
59
~N~ N~NJ~OH
N O ~ ~3 SCH3
N- [2(S)-( 1 -(4-Nitrophenyl-methyl)- 1 H-imidazol-5 -ylacetyl)amino-3 (S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
~NO2
1 ~J
H H
~N~ N N~ N J~OMe
N O ~ ~ SCH3
N-[2(S)-(1 -(4-Cyanophenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-
2 5 methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
~ 5~CN
~N~ N~
N O ~ ~ SCH3
WO g6/10034 PCT/IJS95/12224
1- + .2 '~ ' , ~
22013~8
- 60 -
N- [2(S)-( 1 -(4-Cyanophenyl-methyl) -1 H-imidazol-5-ylacetyl)amino-3 (S )-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine methyl ester
~CN
H H
N~¢ --N~ ~CIMe
15 N-[2(S)-(1-(4-Cyanophenyl-methyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine isopropyl ester
CN
20 ~
H H
~N~ --N~ J~Oi-Pr
2s N O ~ l~3 SCH3
N-[2(S)-(1 -(4-Methoxyphenylmethyl)-lH-imidazol-5-ylacetyl)amino-
3(S)-methylpentyl]-N-l-naphthylmethyl-glycyl-methionine
WO 96tlO034 PCT/US95tl2224
22013~8
- 61 -
OMe
< ~ --N~H
o N-[2(S)-( 1 -(4-Methoxyphenyl-methyl)- 1 H-imidazol-5-ylacetyl)amino-
3(S)-methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methyl ester
OMe
15 ~
< ~ ~N~Me
N-[2(S)-(1 -(2-Naphthylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-methionine
2s
WO g6/10034 PCT/~JS95/12224
,
220=13=~8
- 62 -
~N~ --N~ OH
N O ~ ~3 SCH3
N- [2(S)-( 1 -(2-Naphthylphenyl-methyl)- 1 H-imidazol-5-ylacetyl)amino-
3(S)-methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methyl ester
~
~ O
~N~N N,~ ;
N O ~ ~3 SCH3
N-[2(S)-(1 -(4-Cyanophenyl-methyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-methionine sulfone methyl
ester
WO 96/10034 PCT/US95/12224
~ 2 2 ~ 8 ` ~
.
- 63 -
H H
~N ~ N N~ N J~OMe
N O ~ ~3 SO2CH3
o N-[2(S)-(1-(4-Cyanophenyl-methyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine sulfone
,~,CN
~ H H
~N~N N~NJ~OH
N O ~ ~3 SO2CH3
N-[2(S)-(1 -(4-Cyanophenyl-methyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-2-(acetylamino)~ l~nine methyl
ester
~ H H
3 0 ~N~ N N~ N `~`OMe
- N O ~~ O~CH3
WO 96/10034 . PCT/IJS95112224
22~13~8 ~ --
- 64 -
N-[2(S)-(1 -(4-Cyanophenyl-methyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N- 1 -naphthylmethyl-glycyl-2-(acetylamino)~ l~nine methyl
ester
~,CN
~ H H
~N~N N~NJ~OH
N O --~ O~CH3
N-[2(S)-(1 -(4-Cyanophenylmethyl)-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-N-methyl-methionine
CN
,~
r H ,CH3 O
~N~ N N~N~OH
N O /~ ~3 SCH3
30 N-[2(S)-(1-(4-Cyanophenyl-methyl)-1H-imidazol-5-ylacetyl)amimo-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-N-methyl-methionine methyl
ester
WO 96/10034 PCTIUS95/12224
~ 22~13~8
- 65 -
~CN ICH3 o
~N~ N~N~JJ~oMe
N O ~ ~3 SCH3
or the pharmaceutically acceptable salts thereof.
In the present invention, the amino acids which are
disclosed are identified both by conventional 3 letter and single letter
abbreviations as indicated below:
Alanine Ala A
Arginine Arg R
Asparagine Asn N
Aspartic acid Asp D
Asparagine or
Asparticacid Asx B
Cysteine Cys C
Glllt~mine Gln Q
Glutamic acid Glu E
Glllt~mine or
Glutamic acid Glx Z
Glycine Gly G
Histidine His H
3 0 Isoleucine Ile
Leucine Leu L
Lysine Lys K
Methionine Met M
Phenyl~l~nine Phe F
Proline Pro P
WO 96/10034 PCT/IJS95/12224
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- 66 -
Serine Ser S
Threonine Thr T
Tryptophan Trp W
Tyrosine Tyr Y
Valine Val V
The compounds of the present invention may have
asymmetric centers and occur as racemates, racemic mixtures, and as
o individual diastereomers, with all possible isomers, including optical
isomers, being included in the present invention.
As used herein, "alkyl" is intended to include both
branched and straight-chain saturated aliphatic hydrocarbon groups
having the specified number of carbon atoms.
As used herein, "cycloalkyl" is intended to include non-
aromatic cyclic hydrocarbon groups having the specified number of
carbon atoms. Examples of cycloalkyl groups include cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl and the like.
"Alkenyl" groups include those groups having the
specified number of carbon atoms and having one or several double
bonds. Examples of aLkenyl groups include vinyl, allyl, isopropenyl,
pentenyl, hexenyl, heptenyl, cyclopropenyl, cyclobutenyl,
cyclopentenyl, cyclohexenyl, 1-propenyl, 2-butenyl, 2-methyl-2-
butenyl, isoprenyl, farnesyl, geranyl, geranylgeranyl and the like.
As used herein, "aryl" is intended to include any stable
monocyclic, bicyclic or tricyclic carbon ring(s) of up to 7 members in
each ring, wherein at least one ring is aromatic. Examples of aryl
groups include phenyl, naphthyl, anthracenyl, biphenyl,
tetrahydronaphthyl, indanyl, phen~nthrenyl and the like.
3 0 The term heterocycle or heterocyclic, as used herein,
represents a stable S- to 7-membered monocyclic or stable 8- to 11-
membered bicyclic or stable 11-15 membered tricyclic heterocyclic
ring which is either saturated or unsaturated, and which consists of
carbon atoms and from one to four heteroatoms selected from the
WO 96/10034 PCTIUS95112224
~ 2201348
- 67 -
group consisting of N, O, and S, and including any bicyclic group in
which any of the above-defined heterocyclic rings is ffised to a
benzene ring. The heterocyclic ring may be attached at any
heteroatom or carbon atom which results in the creation of a stable
structure. Examples of such heterocyclic elements include, but are
not limited to, azepinyl, benzimidazolyl, benzisoxazolyl,
benzofurazanyl, benzopyranyl, benzothiopyranyl, benzofuryl,
benzothiazolyl, benzothienyl, benzoxazolyl, chromanyl, cinnolinyl,
dihydrobenzofuryl, dihydro-benzothienyl, dihydrobenzothiopyranyl,
dihydrobenzothio-pyranyl sulfone, furyl, imidazolidinyl,
imidazolinyl, imidazolyl, indolinyl, indolyl, isochromanyl,
isoindolinyl, isoquinolinyl, isothiazolidinyl, isothiazolyl,
isothiazolidinyl, morpholinyl, naphthyridinyl, oxadiazolyl, 2-
oxoazepinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl,
piperidyl, piperazinyl, pyridyl, pyridyl N-oxide, pyridonyl, pyrazinyl,
pyrazolidinyl, pyrazolyl, pyrimidinyl, pyrrolidinyl, pyrrolyl,
quinazolinyl, quinolinyl, quinolinyl N-oxide, quinoxalinyl,
tetrahydrofuryl, tetrahydroisoquinolinyl, tetrahydro-quinolinyl,
thiamorpholinyl, thiamorpholinyl sulfoxide, thiazolyl, thiazolinyl,
thienofuryl, thienothienyl, and thienyl.
As used herein, the terms "substituted aryl",
"substituted heterocycle" and "substituted cycloalkyl" are
intended to include the cyclic group which is substituted with 1
or 2 sub~LiLulents selected from the group which includes but is
not limited to F, Cl, Br, NH2, N(C1-C6 alkyl)2~ CF3, NO2,
(C1-C6 alkyl)O-, -OH, (cl-c6 alkyl)S(O)m-, (C1-C6
alkyl)C(O)NH-, H2N-c(NH)-~ (cl-c6 alkyl)C(O)-, (C1-C6
alkyl)OC(O)-, N3, CN, (cl-c6 alkyl)OC(O)NH- and Cl-c2o
alkyl.
~ .
WO 96/10034 PCT/IJS95112224
220 13g~ - 68 -
The following structure:
,N~
(~H2)t
represents a cyclic amine moiety having S or 6 members in the ring, such
a cyclic amine which may be optionally fused to a phenyl or cyclohexyl
o ring. Examples of such a cyclic amine moiety include, but are not limited
to, the following specific structures:
15 j<r~
When R2a and R2b and R3 and R4 are combined to
form - (CH2)S -, cyclic moieties are formed. Examples of such cyclic
moieties include, but are not limited to:
When RSa and RSb are combined to form - (CH2)s -,
cyclic moieties as described hereinabove for R2a and R2b and R3
and R4 are formed. In addition, such cyclic moieties may optionally
include a heteroatom(s). Examples of such heteroatom-cont~ining
cyclic moieties include, but are not limited to:
WO 96/10034 . PCIIUS95/12224
~ 2201348 - =
- 69 -
5 ~ ~ ~oJ ~sJ
0
Preferably, R1 is selected from: hydrogen, and C1-C6 alkyl.
Preferably, R2a and R2b are independently selected from:
a side chain of a naturally occurring amino acid and C1-C6 alkyl
unsubstituted or substituted with an aryl group.
Preferably, R3 and R4 are independently selected from:
a side chain of a naturally occurring amino acid and C1-C6 alkyl
unsubstituted or substituted with a group selected from aryl, heterocycle
and C3-C1o cycloalkyl.
Preferably, R5a and RSb are independently selected from:
a side chain of a naturally occurring amino acid, methionine sulfoxide,
methionine sulfone and unsubstituted or substituted C1-C6 alkyl .
Preferably, X-Y is selected from:
2s R7b
\55s N~ss and ~55S O~ss
-- =Preferably, R7b C1-C6 alkyl substituted with hydrogen or an
3 o unsubstituted or substituted aryl group.
Preferably, R8 is selected from: hydrogen, perfluoroalkyl, F,
Cl, Br, RlOO-, Rl 1S(O)m-, CN, NO2, R1OC(O)-, R1OOC(O)-,
-N(R10)2, Rl lOC(O)NR10- and Cl-C6 aL~yl.
Preferably, R9 is hydrogen.
WO 96/10034 PCT/IJS95/12224
22013~8 ~
- 70 -
Preferably, R10 is selected from H, C1-C6 alkyl and benzyl.
Preferably, A1 and A2 are a bond.
Preferably, V is selected from hydrogen, heterocycle and
aryl.
Preferably, n, p and r are independently 0, 1, or 2.
Preferably t is 3.
The ph~ ceutically acceptable salts of the compounds
of this invention include the conventional non-toxic salts of the
compounds of this invention as formed, e.g., from non-toxic
o inorganic or organic acids. For example, such conventional non-
toxic salts include those derived from inorganic acids such as
hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoTic, nitric and
the like: and the salts prepared from organic acids such as acetic,
propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric,
15 ascorbic, pamoic, maleic, hydroxymaleic, phenyl-acetic, glutamic,
benzoic, salicylic, sulfanilic, 2-acetoxy-benzoic, fumaric,
toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic,
isethionic, trifluoroacetic and the like.
It is intended that the definition of any substituent or
20 variable (e.g., R10, Z, n, etc.) at a particular location in a molecule
be independent of its definitions elsewhere in that molecule. Thus,
-N(R1032 represents -NHH, -NHCH3, -NHC2H5, etc. It is
understood that substituents and substitution patterns on the
compounds of the instant invention can be selected by one of
25 ordinary skill in the art to provide compounds that are chemically
stable and that can be readily synthesized by techniques known in the
art as well as those methods set forth below.
The ph~ ceutically acceptable salts of the compounds
of this invention can be synthesized from the compounds of this
3 o invention which contain a basic moiety by conventional chemical
methods. Generally, the salts are prepared by reacting the free base
with stoichiometric amounts or with an excess of the desired sal~-
forming inorganic or organic acid in a suitable solvent or various
combinations of solvents.
WO 96110034 . PCT/US95112224
22013~8
- 71 -
The compounds of the invention can be synthesized from
their constituent amino acids by conventional peptide synthesis
techniques, and the additional methods described below. Standard
- methods of peptide synthesis are disclosed, for example, in the
5 following works: Schroeder et al., "The Peptides", Vol. I, Ac~clemic
Press 1965, or Bocl~n~7ky et al., "Peptide Synthesis", Interscience
Publishers, 1966, or McOmie (ed.) "Protective Groups in Organic
Chemistry", Plenum Press, 1973, or Barany et al., "The Peptides:
Analysis, Synthesis, Biology" 2, Chapter 1, Academic Press, 1980, or
Stewart et al., "Solid Phase Peptide Synthesis", Second Edition, Pierce
Chemical Company, 1984. The teachings of these works are hereby
incorporated by reference.
Abbreviations used in the description of the chemistry
and in the Examples that follow are:
Ac2O Acetic anhydride;
Boc t-Butoxycarbonyl;
DBU 1,8-diazabicyclo[5.4.0]undec-7-ene;
DMAP 4-Dimethylaminopyridine;
2 o DME 1,2-Dimethoxyethane;
DMF Dimethylformamide;
EDC 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide-
hydrochloride;
HOBT 1-Hydroxybenzotriazole hydrate;
Et3N Triethyl~mine;
EtOAc Ethyl acetate;
FAB Fast atom bombardment;
HOOBT 3-Hydroxy-1,2,2-benzotriazin-4(3H)-one;
HPLC High-performance liquid chromatography;
MCPBA m-Chloroperoxybenzoic acid;
MsCl Methanesulfonyl chloride;
NaHMDS Sodium bis(trimethylsilyl)amide;
Py Pyridine;
TFA Trifluoroacetic acid;
WO 96/10034 PCT/US95/12224
, = ~ = ~
22013~8
- 72 -
THF Tetrahydr~fu~
Compounds of this invention are prepared by employing
the reactions shown in the following Reaction Schemes A-J, in
addition to other standard manipulations such as ester hydrolysis,
cleavage of protecting groups, etc., as may be known in the literature
or exemplified in the experimental procedures. Some key bond-
forming and peptide modifying reactions are:
Reaction A. Amide bond formation and protecting
group cleavage using standard solution or solid phase methodologies.
Reaction B. Preparation of a reduced peptide subunit by
lreductive aLkylation of an amine by an aldehyde using sodium
cyanoborohydride or other reducing agents.
Reaction C. AIkylation of a reduced peptide subunit
with an alkyl or aralkyl halide or, alternatively, reductive alkylation
of a reduced peptide subunit with an aldehyde using sodium
cyanoborohydride or other reducing agents.
Reaction D. Peptide bond formation and protecting
group cleavage using standard solution or solid phase methodologies.
Reaction E. Preparation of a reduced subunit by borane
reduction of the amide moiety.
These reactions may be employed in a linear sequence to
provide the compounds of the invention or they may be used to
synthesize fragments which are subsequently joined by the alkylation
reactions described in the Reaction Schemes.
REACTION SCHEME A
Reaction A. Couplin~ of residues to form an amide bond
WO 96/10034 PCT/US95/12224
~ 22013~8 ~;
- 73 -
>lO~N I OH + H2N
EDC, HOBT >~ ~ RA H~
or HOOBT HN l ~ OR
Et3N, DMF R
THFCAor r H2N J~OR
REACTION SCHEME B
Reaction B. Preparation of reduced peptide subunits by reductive
alkylation
>~OJ~N~H + ,b,OR
' ~OJ~N N~J~
RB
WO 96/~0034 PCTIUS95/12224
2201348 ; ~
- 74 -
REACTION SCHEME C
Reaction C. Alkylation/reductive alkylation of reduced peptide
subunits
>~O N -J~OR or
RB 1l
RCCH, NaCNBH3
RA R7b
>~O NH ~OR
RB
REACTION SCHEME D
Reaction D. Couplin~ of residues to form an amide bond
~ EDC, HOBT
O RA r 1 or HOOBT
>~OJ~N OH +H2N~ Et3N, DMF
>~oJ~ N ~ N ~J~ HCI or TFA
H O ~J
RA O
H2N Jl~o
~
WO 96/10034 PCT/US95/12224
2201318
- 75 -
REACTION SCHEME E
Reaction E. Preparation of reduced dipeptides from peptides
HN NJ~oR BH3 THF
O RA o
>loJ~ N J~OR
RB
where RA and RB are R2a~ R2b, R3, R4, RSa or R5b as previously
defined; XL is a leaving group, e.g., Br-, I- or MsO-; and RCis
defined such that R7b is generated by the reductive alkylation
process.
Reaction Schemes A-E illustrate bond-forming and peptide
modifying reactions incorporating acyclic peptide units. It is well
20 understood that such reactions are equally useful when the - NHC(RA) -
moiety of the reagents and compounds illustrated is replaced with the
following moiety:
~H2)t
R15
Certain compounds of this invention wherein X-Y is an
3 ethenylene or ethylene unit are prepared by employing the reaction
sequences shown in Reaction Schemes F and G. Reaction Scheme F
outlines the preparation of the alkene isosteres lllili7.ing standard
manipulations such as Weinreb amide formation, Grignard reaction,
acetylation, ozonolysis, Wittig reaction, ester hydrolysis, peptide
WO 96/10034 PCT/US95/12224
22013'i8
- 76 -
coupling reaction, mesylation, cleavage of peptide protecting groups,
reductive alkylation, etc., as may be known in the literature or
exemplified in the Experimental Procedure. The key reactions are:
stereoselective reduction of the Boc-amino-enone to the
5 corresponding syn amino-alcohol (Scheme F, Step B, Part l), alld
stereospecific boron triflouride or zinc chloride activated organo-
magnesio, organo-lithio, or organo-zinc copper(l) cyanide SN2'
displacement reaction (Scheme F, Step G). Through the use of
optically pure N-Boc amino acids as starting material and these two
key reactions, the stereo-chemistry of the final products is well
defined. In Step H of Scheme F, Rx is incorporated using coupling
reaction A and RIcooH; the alkylation reaction C using RXCHO
and a reducing agent; or aLkylation reaction C using RXCH2XL.
The alkane analogs are prepared in a similar m~nner by
including an additional catalytic hydrogenation step as outlined in
Reaction Scheme G.
REACTION SCHEME F
1. ClCO2i-Bu o
BocNHJ~OH MeONHMe BocNH
R2a 2. I R2a
BrMg
Step A
1. NaBH4 OAc
BocNH
2. Ac2O, PY 2
R a
Step B
1. O3, Me2S OAc
2. Ph3P=CHCO2Me BocNH ~CO2Me
Step C R2a
WO 96/10034 PCT/US95/12224
2201348
- 77 -
REACTION SCHEME F (CONT'D)
Step D
1. LiOH OH H MsCI py
BocNH ~ N J~W'
2. EDC, HOBT F~2a ll ~ Step F
amino acid (ester)
W"
W' = OMe, W" = SMe
W'-W"=O
BocNH R3MgCuCNClBF3
R2a O ~ Step G
Wll
1. HCI
R3 H o 2. NaCNBH3
BocNH~ w RXCHO
R2a O ~ Step H
W
R CH2N ~ W'
R2a O W
WO96~0034 PCT/US95112224
,
22013~
- 78 -
REACTION SCHEME F (CONT'D)
NaOH
RXCH2N~ OH
R2a 0 W"
or
1. HCI Alternate
Step H
,!1
2. RXCOH
EDC, HOBT
R3 H O
RX~ N ~J ~ W
R2a 0
W"
NaOH
Rx N~ - OH
O R2a 0
WO 96/10034 PCT/US95/12224
~ 22013~.g ==
,
- 79 -
REACTION SCHEME G
0 1. ClCO2i-Bu o
BocNH J~OH , BocNH
R2a BrMg ~ R2a
1. NaBH4 OAc I 1. O3, Me2S
BocNH ~
2. Ac20, py R2a 2. Ph3P=CHCO2Me
OAc O
BocNH~co Me + H2N"~ 1. LiOH
R2a 1 2. EDC, HOBT
OH H
BocNH ~N~
R2a O -- o MsCI, py
WO 96/10034 PCT/US95112224
22013~
- 80 -
REACTION SCHEME G (CONT'D)
OMs H
BocNH ~N~
- R2a o
1. R3MgCuCNCl-BF3
2. H2, 5% Pd/C
Step K
BocNH~ ,~ O
,
R2a o
1. HCI
2. NaCNBH3,
RXCHO
R3 H
RXCH2NH~ o
R2a o -~
NaOH
R3 H O
RXCH2NH~ OH
R2a O OH
WO 96/10034 PCI/US95/12224
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REACTION SCHEME G (CONT'D)
or 1l
1. HCI 2. RXCOH
EDC, HOBT
xlol R3 H
R CNH~,~b' ~4`0
R2a o
NaOH
1l ~JI~OH
2 0 R2a o OH
The oxa isostere compounds of this invention are
prepared according to the route outlined in Scheme H. An
25 aminoalcohol H-l is acylated with alpha-chloroacetyl chloride in the
presence of trialkyl~mines to yield amide H-2. Subsequent reaction
of H- 2 with a deprotonation reagent (e.g., sodium hydride or
potassium t-butoxide) in an ethereal solvent such as THF provides
morpholinone H-3. The N-Boc derivative H-4 is then obtained by
3 0 the treatment of H-3 with BOC anhydride and DMAP (4-
dimethylaminopyridine) in methylene chloride. Alkylation of H-4
with R3XL, where XL is a leaving group such as Br-, I- or Cl- in
THF/DME (1,2-dimethoxyethane) in the presence of a suitable base,
preferably NaHMDS [sodium bis(trimethylsilyl)amide], affords H-5,
which is retreated with NaHMDS followed by either protonation or
WO 96/10034 PCT/US95112224
3 ~
- 82 -
the addition of an alkyl halide R4X to give H-6a or H-6b,
respectively. Alternatively, H-6a can be prepared from H-4 via an
aldol condensation approach. Namely, deprotonation of H-4 with
NaHMDS followed by the addition of a carbonyl compound
5 RyRzCO gives the adduct H-7 (wherein RY and RZ are selected such
that R3 is eventually provided. Dehydration of H-7 can be effected
by mesylation and subsequent elimin~tion catalyzed by DBU (1,8-
diazabicyclo[5.4.0]undec-7-ene) or the direct treatment of H-7 with
phosphorus oxychloride in pyridine to give olefin H-8. Then,
10 catalytic hydrogenation of H-8 yields H-6a. Direct hydrolysis of H-6
with liLlliulll hydrogen peroxide in aqueous THF will produce acid H-
9b. Sometimes, it is more efficient to carry out this conversion via a
2-step sequence, namely, hydrolysis of H-6 in hydrochloric acid to
afford H-9a, which is then derivatized with BOC-ON or BOC
anhydride to give H-9b. The peptide coupling of acid H-9b with
either an alpha-aminolactone (e.g., homoserine lactone, etc.) or the
ester of an amino acid is carried out under the conditions exemplified
in the previously described references to yield derivative H-10.
Treatment of H-l 0 with gaseous hydrogen chloride gives H- 1 1,
20 which undergoes reductive alkylation in the presence of an aldehyde
RxCHO (H-12) and a reducing agent (e.g., sodium cyanoboro-
hydride); or acylation in the presence of RxCOOH (H-13) and a
peptide coupling reagent affording the products H-14a and_.
Hydrolysis of compounds H-14 to the corresponding hydroxy acids
25 and acids, respectively, is accomplished by standard methods such as
tre~tment with NaOH in alcoholic or aqueous milieux followed by
careful acidifcation with dilute HCl.
WO 96/10034 PCTIUS95/12224
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SCHEME H
sH2N 'R2a ~CI ~ N )"~ Z
H-1 H-2
~~ BOC2O ~ ~
N "' R2a N "' R2a
BOC
H-3 H-4
3 R4
R ~O~ ~
R3X BOC R4HX+or BOC
Base H-5 H-6
RYRZCO
HO~o~ -H20, Rz~O~ 2
N "' R2a N "' R2a
BOC BOC
H-7 H-8
WO 96/10034 PCT/US95/12224
220~3~
- 84 -
SCHEME H (CONT'D)
LiOOH; ,~R4
oraq. HCI, RW-NH o CO2H
then BOC2O R2a
H-~
a RW=H
b, Rw = BOC
EDC R3~R4
9 + H-A ~ BOCNH O I A
R2a o
H-10
HCI R3~R4
HCI NH2 o A
R2a o
H-11
WO 96/10034 PCT/US95/12224
~ 2201348
- ~5 -
SCHEME H (CONT'D)
RXCHO, NaCNBH3 ~ R4
H-12 'RXCH2NH o~A
R2a O
H-11 H-1 4a
RXCOOH, EDC, HOBT
H-13 R3 R4
11 ~"
RXCNH o~A
R2a o
.H-1 4b
A=
J~o or NH J~OR6
~J R5a
q
The thia, oxothia and dioxothia isostere compounds of
this invention are prepared in accordance to the route depicted in
25 Scheme I. Aminoalcohol I-l is derivatized with BOC2O to give I-15.
Mesylation of I-15 followed by reaction with methyl alpha-
mercaptoacetate in the presence of cesium carbonate gives sulfide I-
16. Removal of the BOC group in I-16 with TFA followed~by
neutralization with di-isopropylethyl~mine leads to lactam I-17. N-
30 BOC derivative I-18 is obtained via the reaction of I-17 with BOC
anhydride in THF catalyzed by DMAP. Sequential alkylation of I- 1
with the alkyl halides R3X and R4X in THF/DME using NaHDMS
as the deprotonation reagent produces I-l 9. Hydrolysis of I-l9 in
hydro-chloride to yield I-20a, which is derivatized with Boc
WO 96/10034 PCr/US95/12224
. . . --
~?,o~3~
- 86 -
anhydride to yield I-20b. I~he coupling of I-20b with an alpha-
aminolactone (e.g., homoserine lactone, etc.) or the ester of an amino
acid is carried out under conventional conditions as exemplified in
the previously described references to afford I-21. Sulfide I-21 is
5 readily oxidized to sulfone I-22 by the use of MCPBA (m-
chloroperoxybenzoic acid). The N-BOC group of either I-21 or I-22
is readily removed by treatment with gaseous hydrogen chloride.
The resultant amine hydrochloride I-23 undergoes reductive
aL~ylation in the presence of an aldehyde RXCHO (I-l2) and a
reducing agent (e.g., sodium cyanoborohydride); or acylation in the
presence of RxcooH (I-13) and a peptide coupling reagent to afford
the products I-24 and I-25.
WO 96/10034 PCTIUS95/12224
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SCHEME I
HO~ HO
1 ) MsCI
BOC20 HN ~"' 2 2) Cs2CO3
l 1S 1-15 BOC HSCH2C02CH3
CH32C HN~"' R2a 1 ) TFA ~ ~) BOC20
2) (i-Pr)2- N "~R2a
1-16 BOC NEt
-- H 1 17
R4
~S~ 1 ) R3X, R3"~ ~ HCI
O 1 2) R4X, ~ J"~ 2 H20 '
1-18 BOC Base 1-19 BOC
Nl H R~R4 H-A, EDC BOCNH R~A
Rw R2a HOBT R2a O
1-21
1-20
a, RW=H ~ BOC20 NHJ~o or NH~I~ 6
b, RW=BOC ~ R5a
WO 96/10034 PCT/US9',/12224
~0~ 4
- 88 -
SCHEME I (CONT'D)
R ~R
BOCNH S(O)?~A HCI
R2a O
m=0, 1-20 ~ MCPBA
m=2, 1 22
R3 R4 R ~R
R2a ( )~ RXCHO R2a X
12
1-23 NaCNBH3 1-24
m = 0 or 2
R1COOH
EDC, HOBT
2 5 R~CNH R3?~A
R2a o
3 0 1-25
WO 96tlO034 2 2 0 1 3 4 8 PCT/US95/12224
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Reaction Schemes J - M illustrate reactions wherein the non-
sulfhydryl-cont~ining moiety at the N-termiml~ of the compounds of the
5 instant invention is attached to an acyclic peptide unit which may be
further elaborated to provide the instant compounds. These reactions
may be employed in a linear sequence to provide the compounds of the
invention or they may be used to synthesize fragments which are
subsequently joined by the reactions described in Reaction Schemes A -
10 E-
The intermediates whose synthesis are illustrated in Reaction
Schemes A and C can be reductively alkylated with a variety of
aldehydes, such as V, as shown in Reaction Scheme J. The aldehydes
can be prepared by standard procedures, such as that described by O. P.
Goel, U. Krolls, M. Stier and S. Kesten in Or~anic Syntheses, 1988, 67,
69-75, from the a~lol~liate amino acid (Reaction Scheme J). The
reductive alkylation can be accomplished at pH 5-7 with a variety of
reducing agents, such as sodium triacetoxyborohydride or sodium
cyanoborohydride in a solvent such as dichloroethane, methanol or
2 dimethylformamide. The product VI can be deprotected to give the final
compounds VII with trifluoroacetic acid in methylene chloride. The final
product VII is isolated in the salt form, for example, as a trifluoroacetate,
hydrochloride or acetate salt, among others. The product ~ mine VII
can further be selectively protected to obtain VIII, which can
25 subsequently be reductively alkylated with a second aldehyde to obtain
IX. Removal of the protecting group, and conversion to cyclized
products such as the dihydroimidazole XI can be accomplished by
literature procedures.
Alternatively, the protected dipeptidyl analog intermediate
can be reductively alkylated with other aldehydes such as l-trityl-4-
carboxaldehyde or l-trityl-4-imidazolylacetaldehyde, to give products
such as XII (Reaction Scheme K). The trityl protecting group can be
removed from XII to give XIII, or alternatively, XII can first be treated
with an alkyl halide then subsequently deprotected to give the alkylated
WO 96/10034 PCI/US95/12224
1 --
2?,0~3~'8 ,
- 90 -
imidazole XIV. Alternatively, the dipeptidyl analog intermediate can be
acylated or sulfonylated by st~ndard techniques.
The imidazole acetic acid XV can be converted to the
acetate XYII by standard procedures, and XVII can be first reacted with
5 an alkyl halide, then treated with refluxing methanol to provide the
regiospecifically aL~ylated imidazole acetic acid ester XVIII. Hydrolysis
and reaction with the protected dipeptidyl analog intermediate in the
presence of condensing reagents such as 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide (EDC) leads to acylated products such as X~X.
Similar procedures as are illustrated in Reaction Schemes J-
M may be employed using other peptidyl analog intermediates such as
those whose synthesis is illustrated in Reaction Schemes B - I.
wo96/10034 PCrlUSg5/12224
2201348
- 91 -
REACTION SCHEME J
Y RB Boc NHl V
2 ~N I OR Boc NH CHO
RA H O NaBH(OAc)3
Et3N, CICH2CH2CI
NHBoc
/~ H Y RB CF3CO2H
Boc NH \--N~J~`N~OR CH2C12
RA H O
Vl
NH2
~_H Y RB Boc20
NH2 N~J`N OR CH2CI2
Vll
2s BocNH/~N~JI~N I OR CHO
- A H NaBH(OAc)3
R Et3N, CICH2CH2CI
Vlll
WO 96/10034 PCT/US95112224
2~,0~3 1~
- 92 -
REACTION SCHEME J (continued)
5 ~
NH RB CF3CO2H, CH2CI2;
BocNH/~ NH\,J~ ~ NaHCO3
RA H o
~=~ IX
~,
NH
~ ~J~N ~NC
RA H o AgCN
X
/~ RA H~
N~N~
2s ~3 Xl
-
WO 96/10034 2 2 0 1 3 ~ 8 PCT/US95/12224
.
- 93 -
REACTION SCHEME K
H2N~J~ OR NaBH(OAc)3
-A NH Et3N, CICH2CH2CI
N~(CH2)nCHO
~NT~
o H Y RB
~CH2)n-- JJ~N~OR
N~ RA H O
,N
Tr Xll 1 ) Ar CH2X, CH3CN
2) CF3CO2H, CH2CI2
CF3CO2H, CH2CI2 (C2H5)3siH
(C2H5)3SiH
H2)n+1 -~ N~
N~ RA H O
~N~
H Xlll
H Y RB
Ar ~CH2)~ J~N~R
N
XIV
WO 96110034 PCI/US95/12224
3 4S
- 94 -
REACTION SCHEME L
N~ H2C02H CH 0 N~CH2C02CH3
H HCI NH HCI
XV XVI
(c6H5)3cBr N~ 2 02CH31 ) ArCH2X CH3CN
o (C2DH )3FN ,N 2) CH30H, reflux
XVII
Ar--\N~CH2co2cH3 2.5N HClaq,
N 55C
XVIII
Ar~\N CH2CO2H
WO 96110034 2 2 0 1 ~ 4 8 PCT/US95/12224
_ 95 _
REACTION SCHEME M
s ~ + H2N~J~N~,O~
EDC HCI
HOBt
DMF
Ar~ ~N~OR
XIX
WO 96/10034 . , PCT/US95112224
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The compounds of this invention inhibit Ras farnesyl
transferase which catalyzes the first step in the post-translational
processing of Ras and the biosynthesis of functional Ras protein.
These compounds are useful as ph~rm~ceutical agents for m~mm~
5 especially for humans. These compounds may be ~lmini~tered to
patients for use in the treatment of cancer. Examples of the type of
cancer which may be treated with the compounds of this invention
include, but are not limited to, colorectal carcinoma, exocrine
pancreatic carcinoma, and myeloid leukemias.
The compounds of this invention are also useful for
inhibiting proliferative diseases, both benign and malignant, wherein Ras
proteins are aberrantly activated as a result of oncogenic mutation in
other genes (i.e., the Ras gene itself is not activated by mutation to an
oncogenic form) with said inhibition being accomplished by the
~lmini.~tration of an effective amount of the compounds of the invention
to a m~mm~l in need of such treatment. For example, the benign
proliferative disorder neurofibromatosis, or tumors in which the Ras is
activated due to mutation or overexpression of tyrosine kinase oncogenes
(e.g., neu, src, abl, lck, and fyn) may be inhibited by the compo~mds of
20 this invention. Furthermore, arteriosclerosis and diabetic disturbance of
blood vessels may be prevented or treated by use of the instant
compounds to inhibit proliferation of vascular smooth muscle cells.
The compounds of this invention may be ~clmini~tered
to m~mm~l~, preferably hllm~n~, either alone or, preferably, in
25 combination with ph~rm~ceutically acceptable carriers or diluents,
optionally with known adjuvants, such as alum, in a ph~rm~ceutical
composition, according to standard ph~rm~ceutical practice. The
compounds can be ~tlmini.~tered orally or parenterally, including the
intravenous, intramuscular, intraperitoneal, subcutaneous, rectal and
30 topical routes of ~tlmini~tration.
For oral use of a chemotherapeutic compound according
to this invention, the selected compound may be ~lministered, for
example, in the form of tablets or capsules, or as an aqueous solution
or suspension. In the case of tablets for oral use, carriers which are
WO 96/10034 PCT/US9~/12224
22Q1~18
- 97 -
commonly used include lactose and corn starch, and lubricating
agents, such as magnesium stearate, are commonly added. For oral
~lmini~tration in capsule form, useful diluents include lactose and
dried corn starch. When aqueous suspensions are required for oral
use, the active ingredient is combined with emulsifying and
suspending agents. If desired, certain sweetening and/or flavoring
agents may be added. For intramuscular, intraperitoneal,
subcutaneous and intravenous use, sterile solutions of the active
ingredient are usually prepared, and the pH of the solutions should be
suitably adjusted and buffered. For intravenous use, the total
concentration of solutes should be controlled in order to render the
preparation isotonic.
The present invention also encompasses a
pharmaceutical composition useful in the treatment of cancer,
comprising the administration of a therapeutically effective amount
of the compounds of this invention, with or without pharmaceutically
acceptable carriers or diluents. Suitable compositions of this
invention include aqueous solutions comprising compounds of this
invention and pharmacologically acceptable carriers, e.g., saline, at a
pH level, e.g., 7.4. The solutions may be introduced into a patient's
intramuscular blood-stream by local bolus injection.
When a compound according to this invention is
~lmini~tered into a human subject, the daily dosage will normally be
determined by the prescribing physician with the dosage generally
varying according to the age, weight, and response of the individual
patient, as well as the severity of the patient's symptoms.
In one exemplary application, a suitable amount of
compound is ~lministered to a m~mm~l undergoing treatment for
cancer. A11mini~tration occurs in an amount between about 0.1
mg/kg of body weight to about 20 mg/l~g of body weight per day,
preferably of between 0.5 mg/kg of body weight to about 10 mg~g
of body weight per day.
The compounds of the instant invention are also useful as
a component in an assay to rapidly determine the presence and
WO 96/10034 PCTIUS95/12224
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quantity of farnesyl-protein transferase (FPTase) in a composition.
Thus the composition to be tested may be divided and the two
portions contacted with n~ res which comprise a known substrate
of FPTase (for example a tetrapeptide having a cysteine at the amine
le",li""~) and farnesyl pyrophosphate and, in one of the mixtures, a
compound of the instant invention. After the assay mixtures are
incubated for an sufficient period of time, well known in the art, to
allow the FPTase to farnesylate the substrate, the chemical content of
the assay mi~Lur~s may be determined by well known immunological,
radiochemical or chromatographic techniques. Because the
compounds of the instant invention are selective inhibitors of Fl'Tase,
absence or qll~ntit~tive reduction of the amount of substrate in the
assay mixture without the compound of the instant invention relative
to the presence of the unchanged substrate in the assay cont~ining the
instant compound is indicative of the presence of FPTase in the
composition to be tested.
It would be readily apparent to one of ordinary skill in the
art that such an assay as described above would be useful in identifying
tissue samples which contain farnesyl-protein transferase and quantitating
the enzyme. Thus, potent inhibitor compounds of the instant invention
may be used in an active site titration assay to determine the quantity of
enzyme in the sample. A series of samples composed of aliquots of a
tissue extract cont~ining an unknown amount of farnesyl-protein
transferase, an excess amount of a known substrate of FPTase (for
example a tetrapeptide having a cysteine at the amine terminus) and
farnesyl pyrophosphate are incubated for an a~lo~-iate period of time in
the presence of varying concentrations of a compound of the instant
invention. The concentration of a sufficiently potent inhibitor (i.e., one
that has a Ki subst~nti~lly smaller than the concentration of enzyme in the
assay vessel) required to inhibit the enzymatic activity of the sample by
50% is approximately equal to half of the concentration of the enzyme in
that particular sample.
WO 96/10034 PCT/US95/12224
~ 2 ~
99
EXAMPLES
Examples provided are intended to assist in a further
understanding of the invention. Particular materials employed,
5 species and conditions are intended to be further illustrative of the
invention and not limitative of the reasonable scope thereof.
The standard workup referred to in the examples refers
to solvent extraction and washing the organic solution with 10%
citric acid, 10% sodium bicarbonate and brine as appropriate.
0 Solutions were dried over sodium sulfate and evaporated in vacuo on
a rotary evaporator.
EXAMPLE 1
15 Preparation of N-[2(S)-(1 -(Phenylmethyl)-lH-imidazol-4-yl-
acetyl)amino-3(S)-methylpentyl]-N-1-naphthylmethyl-glycyl-methionine
bis trifluoroacetate (13) and N-[2(S)-(1-(Phenylmethyl)-lH-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl] -N- 1 -naphthylmethyl-glycyl-
methionine bis trifluoroacetate (14)
Step A: Preparation of lH-Imidazole-4-acetic acid methyl ester-
hydrochloride (I)
Into a solution of lH-imidazole-4-acetic acid hydrochloride
(4 g, 24.6 mmol) in methanol (100 ml) was bubbled hydrogen chloride
25 gas until saturated. This solution was allowed to stand for 18 h at room
temperature and the solvent evaporated in vacuo to give (1) as a white
solid.
1H NMR (CDCl3, 400 MHz) o 8.85 (lH, s), 7.45 (lH, s), 3.89 (2H, s)
and 3.75 (3H, s) ppm.
Step B: Preparation of 1 -(Phenylmethyl)-lH-imidazol-4-ylacetic
acid methyl ester (2) and 1-(Phenylmethyl)-lH-imidazol-5-
ylacetic acid methyl ester (3) (3:1 mixture)
WO 96/10034 PCIIUS95/12224
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- 100-
To a solution of sodium hydride (37.3 mg, 1.56 mnnol) in
dimethylform~mide (2 ml) cooled to 0C (ice bath) was added, via
c~nn~ , a solution of 1 (115 mg, 0.707 mmol) in dimethylformamide (3
ml). This suspension was allowed to stir at 0C for 15 min. To this
suspension was added benzyl bromide (84 ~L, 0.707 mmol) andL the
mixture was stirred at room temperature for 2h. After this time, the
mixture was quenched with sat. aq. sodium bicarbonate (15 ml) and water
(20 ml) and extracted with methylene chloride (2 x 50 ml). The
combined extracts were washed with brine (20 ml), dried (MgSO4),
filtered and the solvent was evaporated in vacuo. The residue was
purified by flash chromatography eluting with acetonitrile to give a 3: l
mixture of 2 and 3.
1H NMR (CDC13, 400 MHz) ~ 7.53 (0.25H, s), 7.48 (0.75H, s), 7.35
(3H,m), 7.18 (l.SH, d, J=7.4 Hz), 7.06 (0.5H, d, J=7.2 Hz), 7.00 (0.25H,
s), 6.87 (0.75H, s), 5.16 (O.SH, s), 5.08 (1.5H, s), 3.72 (1.5H, s), 3.65
(2.25H, s), 3.63 ~.75H, s) and 3.48 (0.5H, s) ppm.
Step C: Preparation of 1-(Phenylmethyl)-lH-imidazol-4-ylacetic
acid hydrochloride (4) and 1-(Phenylmethyl)-lH-inaidazol-
5-vlacetic acid hydrochloride (5) (3: lmixture)
A solution of 2 and 3 (3: 1 mixture, 106 mg) in 1.0 N HCl (3
ml) was heated to 45C for 4 h. After this time, the solution was
evaporated in vacuo to give a 3:1 mixture of 4 and 5.
1H NMR (DMSO, 400 MHz) ~ 9.26 (0.75H, s), 9.23 (0.25H, s), 7.60
(0-25H, m), 7.58 (0.75H, s), 7.45-7.26 (5H, m), 5.43 (0.5H, s), 5.41
(0.5H, s), 3.77 (1.5H, s), 3.75 (0.5H, s) ppm.
Step D: Preparation of N-(2(S)-(t-butoxycarbonylamino)-3(S)-
methylpentyl)glycine methyl ester (6)
Glycine methyl ester hydrochloride (4.41 g, 0.035 mol) was
dissolved in 1,2-dichloroethane (50 mL) and DMF (5 mL) and treated
with 3A molecular sieves (10 g) and N-t-butoxycarbonyl-isoleucinal (6.3
g, 0.029 mol) with stirring at 0C. Sodium triacetoxyborohydride (9.27
g, 0.044 mol) was added, and the pH of the mixture was adjusted to 6
with triethylamine (3 mL, 0.022 mol). After stirring for 18 h the mixture
WO 96/10034 PCI~/US95/12224
~ 22013~8
- 101 -
was filtered, concentrated to a small volume and partitioned between
EtOAc and water. The aqueous layer was extracted with EtOAc (2 x 50
mL). The combined organic phase was washed with aqueous saturated
NaHCO3 solution, brine, and dried (Na2SO4). Filtration and
5 concentration afforded a residue which was purified by flash
chromatography (siO2, EtOAc:hexane, 1 :3) to give (6).
1H NMR (CDCl3) ~ 4.69 (lH, m), 3.72 (3H, s), 3.48-3.62 (lH, m), 3.42
(2H, ABq), 2.65 (2H, d, J=6 Hz), 1.4-1.6 (2H, m), 1.48 (9H, s), 1.04-1.2
(lH, m), 0.85-0.95 (6H, m) ppm.
Step E: Preparation of N-[2(S)-(t-Butoxycarbonylamino)-3(S)-
methylpentyll-N-(l-naphthylmethyl)~lycine methyl ester (7)
N-[2(S)-(t-Butoxycarbonylamino)-3(S)-methyl-pentyl]-
glycine methyl ester (6, 2.00 g, 6.97 mmol) was dissolved in 1,2-
dichloroethane (56 ml) and 3A molecular sieves were added followed by
l-naphthaldehyde (1.89 ml, 13.9 mmol) and sodium triacetoxy-
borohydride (6.65 g, 31.4 mmol). The mixture was stirred at ambient
temperature for 16 h, and filtered through glass fiber paper and
concentrated. The residue was partitioned between EtOAc and sat.
20 NaHCO3 (100 ml/25 ml). The aqueous layer was extracted with EtOAc
(3xS0 ml). The organic layers were combined, dried (Na2so4)~ filtered,
and concentrated to give 5.0 g of crude product which was purified by
chromatography (sio2~ 15-33% ethyl acetate/hexane) to give 7 .
1H NMR (CD30D) ~ 8.44-8.38 (lH, d, J=6Hz), 7.88-7.77 (2H, m,),
25 7.55-7.35 (4H, m), 6.34-6.27 (lH, m), 4.25 (2H, ABq), 3.66 (3H, s),
3.40-3.23 (lH, m), 2.90 (lH, dd, J=6 and 15Hz), 2.63 (lH, dd, J=6 and
15Hz), 1.57-1.46 (lH, m), 1.43 (9H, s), 1.34-1.18 (2H, m), 1.06-0.85
(lH, m) and 0.85-0.71 (6H, m) ppm.
30 Step F: Preparation of N-[2(S)-(t-Butoxycarbonylamino)-3(S)-
methylpentyll-N-(l-naphthylmethyl)glycine (8)
N-[2(S)-(t-Butoxycarbonylamino)-3(S)-methylpentyl]-N-(l -
naphthylmethyl)glycine methyl ester (7, 2.61 g, 6.10 mmol) was
dissolved in MeOH (50 ml) and lN NaOH (24.4 ml, 24.4 mmol) was
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added. The mixture was stirred at ambient temperature for 4 h and
concentrated. The resulting residue was dissolved in water (25 ml) and
neutralized with lN HCI (24.4 ml). The aqueous layer was washed with
EtOAc (3x50 ml). The organic layers were combined, dried with
5 Na2SO4, filtered, and concentrated to give the product. lH NMR
(CD30D) ~ 8.43 (lH, d, J=6Hz), 7.97 (2H, t, J=6 Hz) 7.75-7.48 (4H, m),
4.96 (lH, d, J=12Hz), 4.72 (lH, d, J=12 Hz), 3.80-3.58 (3H, m), 3.49-
3.40 (lH, dd" J=3 and 12 Hz), 3.03 (lH, dd, J=3 and 12 Hz), 1.42 (9H,
s,), 1.37-1.28 (2H, m), 1.80-1.00 (lH, m), 0.94-0.78 (6H, m,) ppm.
Step G: Preparation of N-[2(S)-(t-Butoxycarbonylamino)-3(S)-
methylpentyl]-N-(1 -naphthylmethyl)glycine-methionine
methyl ester (9)
N-[2(S)-(t-Butoxycarbonylamino)-3(S)-methylpentyl~-N-( l -
15 naphthylmethyl)glycine (8, 2.29g, 5.53 mmol), dissolved in DMF (20mL), was treated with HOBT (0.822 g, 6.08 mmol), EDC (1.17 g, 6.0~s
mmol), and methionine methyl ester hydrochloride (1.21 g, 6.08 mmol).
The pH was adjusted to 7.5 with Et3N (1.7 mL, 12 mmol) and the
mixture was stirred at ambient temperature for 24 h. The mixture was
20 concentrated, and the residue was partitioned between EtOAc (50 mL)
and saturated NaHCO3 solution (25 mL). The aqueous layer was
extracted with EtOAc (lx30 mL). The organic layers were combined,
washed with brine (lx25 mL), dried (Na2S04), ~lltered, and concentrated
to give 3.2 g of crude product which was purified by chromatography
25 (silica gel eluting with 1 :3 to 1 :2 ethyl acetate in hexane) to give pure
product. 1H NMR (CD30D) ~ 8.33 (lH, d, J=6 Hz), 7.90 (lH, d, J=6
Hz), 7.82 (1H, d, J=6 Hz), 7.61-7.39 (4H, m), 6.60-6.52 (lH, m), 4.32-
4.06 (2H, m), 3.90-3.69 (lH, m), 3.65 (3H, s), 3.27-3.14 (2H, m), 2.93-
2.70 (2H, m), 2.19-1.78 (6H, m), 1.63-1.30 (13H, m), 1.19-1.05 (lH, m),
30 0.95-0.81 (6H, m) ppm.
Step H: Preparation of N-(2(S)-amino-3(S)-methylpentyl)-N-(1-
naphthylmethyl)-glycyl-methionine methyl ester
hydrochloride (10)
.
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N-[2(S)-(t-Butoxycarbonylamino)-3 (S)-methylpentyl] -N-(l -
naphthylmethyl)-glycyl-methionine methyl ester (9, 2.82 g, 5.04 mmol)
was dissolved in EtOAc (50 mL) and cooled to -25C. HCl was bubbled
through the mixture until TLC (95:5 CH2Cl2:MeOH) indicated complete
reaction. Nitrogen was bubbled through the mixture to remove excess
HCl and the mixture was then concentrated to give the title compound.
1H NMR (CD30D) ~ 8.31 (lH, d, J=6 Hz), 7.96 (2H, d, J=6 Hz), 7.g3-
7.71 (lH, m), 7.68-7.49 (3H, m), 4.76-4.55 (4H, m), 3.84-3.75 (2H, m),
3.71 (3H, s), 3.70-3.59 (lH, m), 3.21-3.00 (2H, m), 2.57-2.38 (3H, m),
2.17-2.04 (4H, m), 1.97-1.81 (lH, m), 1.63-1.50 (lH, m), 1.39-1.20 (lH,
m), 1.19-1.00 (lH, m), 0.95-0.79 (6H, m) ppm.
Step I: Preparation of N-[2(S)-(l-(Phenylmethyl)-lH-imidazol-4-
ylacetyl)amino-3(S)-methylpentyl] -N- 1 -naphthylmethyl-
gylycyl-methionine methyl ester bis trifluoroacetate (11) and
N-[2(S)-(1-(Phenylmethyl)-lH-imidazol-5-ylacetyl)-amino-
3(S)-methylpentyl]-N-1-naphthylmethyl-glycyl-methionine
methyl ester bis trifluoroacetate tl2)
To a solution of a l-(phenylmethyl)-lH-imidazol-4-ylacetic
acid hydrochloride (4) and 1 -(phenylmethyl)-lH-imidazol-5-ylacetic acid
hydrochloride (5, 3:1 mixture, 115 mg, 0.455 mmol), N-[2(S)-amino-
3(S)-methylpentyl]-N-1-naphthylmethyl-glycyl-methionine methyl ester
bis hydrochloride (10, 244 mg, 0.455 mmol) and 3-hydroxy-1,2,3-
benzotriazin-4(3H)-one (HOOBT, 74 mg, 0.46 mmol) in
dimethylform~mide (5 ml) was added 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (EDC, 87 mg, 0.455 mmol) and
triethyl~mine (190 ,ul, 1.36 mmol) and the solution stirred overnight.
After this time, sat. aq. sodium bicarbonate (20 ml) and water (25 ml)
were added and the mixture was extracted with ethyl acetate (2 X 50 ml).
The combined extracts were washed with brine (5 ml) and the solvent
evaporated in vacuo. The regioisomers were separated by Prep HPLC
using a Nova Prep 5000 Semi preparative HPLC system and a Waters
PrepPak cartridge (47 X 300mm, C18, 15 um, 100A) eluting with 5 -
WO 96/10034 PCTJUS95/12224
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95% acetonitrile/water (0.1% TFA) at 100 ml/min (chromatography
method A) to give after lyophili7~tion pure 11 and 12.
11:
lH NMR (CD30D, 400MHz) ~ 8.95 (lH, s), 8.27 (lH, m), 7.96 (2H,
m), 7.68 (lH, d), 7.60-7.37 (9H, m), 5.38 (2H, s), 5.0-4.8 (lH, m), 4.52
(lH, t, J=10.6 Hz), 4.42 (lH, dd, J=4 and 6.6 Hz), 4.14 (lH, m), 3.92
(lH, d, J=13.3 Hz), 3.83 (lH, d, J=13.3 Hz), 3.70 (lH, s), 3.64 (lH, m),
3.54 (2H, m), 3.22 (lH, dd, J=7 and 8 Hz), 2.37 (lH, m), 2.10 ~lH, m),
2.00 (3H, s), 1.98 (lH, m), 1.79 (lH, m), 1.58 (lH, m), 1.42 (lH, m),
1.17 (lH, m) and 0.90 (6H, m) ppm.
Anal. Calcd for C37H47N504S-3.0 TFA-0.15 H2O: C, 51.51; H, 5.06,
N, 6.98. Found: C, 51.52; H, 4.98; N, 7.18.
FAB HRMS exact mass calcd for C37H4gN5O4S 658.342702 (MH+),
found 658.341278.
12:
lH NMR (CD30D, 400 MHz) ~ 8.8 (lH, s), 8.26 (lH, m), 7.89 (2H, m),
7.66-7.24 (8H, m), 7.21 (2H, s), 5.36 (2H, m), 4.37 (3H, m), 4.09 (lH,br
s), 3.66 (3H, s), 3.56 (3H, m), 3.50-2.90 (3H, m), 2.27 (lH, br s), 2.20
(lH, br s), 1.96 (3H, s), 1.90 (lH, br s), 1.6~S (lH, br s), 1.58 (lH, br s),
1.40 (lH, m), 1.18 (lH, m) and 0.89 (6H, m~ ppm.
Anal. Calcd for C37H47NsO4S-1.85 TFA-0.10 H20: C, 56.15, H, 5.68;
N, 8.04. Found: C, 56.14; H, 5.62; N, 8.44.
FAB HRMS exact mass calcd for C37H4gN5O4S 658.342702 (MH+),
found 658.343754.
Step J: Preparation of N-[2(S)-(l-(Phenylmethyl)-lH-imidazol-4-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
glycyl-methionine bis trifluoroacetate (13) and N-~2(S)-(l-
(Phenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-l-naphthylmethyl-glycyl-methionine bis
trifluoroacetate (14)
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To a solution of N-[2(S)-(l-(Phenylmethyl)-lH-imidazol-4-
ylacetyl)amino-3(S)-methylpentyl]-N- l -naphthylmethyl-glycyl-
methionine methyl ester bis trifluoroacetate (11) and N-[2(S)-(l-
(phenylmethyl)- 1 H-imidazol-5 -yl)acetyl)amino-3 (S )-methylpentyl] -N- 1 -
5 naphthylmethyl-glycyl-methionine me~yl ester bis trifluoroacetate (12,
2: 1 mixture, 50 mg, 0.057 mmol) in methanol ( 5 ml ) was added l .ON
lithium hydroxide (570 ,ul, 0.547 mmol ). This solution was stirred for 4 h
and treated with trifluoroacetic acid (100 ~l). This mixture was purified
by preparative HPLC using chromatography method A to give the title
1 o compounds.
13:
lH NMR (CD30D, 400 MHz) ~ 8.83 (lH, s), 8.21 (lH, d, J=9.5 Hz),
7.88 (2H, m), 7.54 (lH, d, J=6.9 Hz), 7.5 - 7.3 (9H, m), 5.32 (2H, s), 4.56
15 (lH, br d, J = 10 Hz), 4.36 (2H, m), 4.09 (lH, m), 3.55 (4H, m), 3.17
(lH, br d, J = 10 Hz), 2.98 (lH, t, J = lOHz), 2.29 (lH, m), 2.18 (lH, m),
1.96 (lH, m), 1.95 (3H, s), 1.67 (lH, m), 1.56 (lH, m), 1.37 (lH, m),
1.11 (lH, m) and 0.88 (6H, m) ppm.
Anal. Calcd for C36H45N504S-2.15 TFA: C, 54.45; H, 5.35; N, 7.P~8.
20 Found: C, 54.42; H, 5.30; N, 7.97.
FAB HRMS exact mass calcd for C36H46N504S 644.327052 (MH+),
found 644.326691.
.
14:
25 lH NMR (CD30D, 400 MHz) ~ 8.80 (lH, s), 8.29 (lH, m), 7.92 (2H,
m), 7.61 (lH, br), 7.32-7.53 (7H, m), 7.21 (2H, br s), 5.37 (2H, s), 4.37
(2H, m), 4.08 (lH, m), 3.57 (4H, br m), 3.05 (2H, m), 2.29 (2H, m), 2.20
(lH, m), 1.96 (3H, s), 1.70 (lH, m), 1.62 (lH, m), 1.57 (lH, m), 1.39
(lH,m), 1.13 (lH,m)andO.88 (6H,m)ppm.
30 FAB HRMS exact mass calcd for C36H46N504S 644.327052 (MH+),
found 644.327917.
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EXAMPLE 2
Preparation of N-[2(S)-(1-(4-Nitrophenylmethyl)-lH-irnidazol-4-
ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -naphthylmethyl-glycyl-
5 methionine bis trifluoroacetate (21) and N-[2(S)-(1-(4-
Nitrophenylmethyl)- 1 H-imidazol-5-ylacetyl)amino-3(S)-methylpentyl] -
N-1-naphthylmethyl-glycyl-methionine bis trifluoroacetate (22).
Step A: Preparation of 1-(4-Nitrophenylmethyl)-lH-imidazol-4-
ylacetic acid methyl ester (15) and 1-(4-NitrophenyLrnethyl)-
lH-imidazol-5-ylacetic acid methyl ester (16) (3:1mixture)
To a solution of sodium hydride (60% in mineral oil, 99 mg,
2.5 mmol) in dimethylforrn~mide (2 ml) cooled to 0C was added, via
c~nn~ , a solution of lH-imidazole-4-acetic acid methyl ester
hydrochloride (1, 200 mg, 1.13 mmol) in dimethylformamide (3 ml).
This suspension was allowed to stir at 0C for 15 min. To this
suspension was added 4-nitrobenzyl bromide (244 mg, 1.13 mmol) and
stirred at room temperature for 2 h. After this time, the mixture was
quenched with sat. aq. .sodium bicarbonate (15 ml) and water (20 ml) and
extracted with methylene chloride (2 x 50 ml). The combined organic
extracts were washed with brine (20 ml), dried (MgSO4), filtered and the
solvent was evaporated in vacuo. The residue was purified by flash
chromatography using acetonitrile as eluent to give the title compounds
as a yellow oil.
1H NMR (CDC13, 400 MHz) ~ 8.20 (2H, d, J=8.5 Hz), 7.49 (lH, s), 7.27
(2H, d, J=8.5 Hz), 7.03 (0.25H, s), 6.87 (0.75H, s~, 5.28 (O.SH, s), 5.18
(l.SH, s), 3.70 (2.25H, s), 3.65 (l.SH, s), 3.61 (0.75H, s) and 3.44 (O.SH,
s) ppm.
30 Step B: Preparation of 1-(4-Nitrophenylmethyl)-lH-imidazol-4-
ylacetic acid hydrochloride (17) and 1-(4-Nitrophenyl-
methyl)-lH-imidazol-5-vlacetic acid tl8) (3:1mixture)
To a solution of a mixture of 1-(4-Nitrophenylmethyl)-lH-
imidazol-4-ylacetic acid methyl ester (15) and 1-(4-Nitrophenylmethyl)-
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lH-imidazol-5-ylacetic acid methyl ester (16, 3:1mixture, 216 mg, 0.785
mmol) in methanol (3 ml) and tetrahydrofuran (3 ml) under argon was
added 1.0 M sodium hydroxide (1.18 ml, 1.18 mmol) and stirred for 18 h.
After this time, 1.0 N hydrochloric acid (2.36 ml, 2.36 mmol) was added
and the mixture evaporated in vacuo to give the title compounds.
lH NMR (CDCl3, 400 MHz) o 9.04 (0.75H, s), 8.83 (0.25H, s), 8.28
(2H, d, J=8.8 Hz), 7.61 (2H, d, J=8.8 Hz), 7.54 (0.75H, s), 7.43 (0.25H,
s), 5.61 (0.5H, s), 5.58 (1.5H, s), 3.84 (0.5H, s) and 3.82 (1.5H, s) ppm.
Step C: Preparation of N-[(2S)-(1-(4-Nitrophenylmethyl)-lH-
imidazol-4-ylacetyl)amino-3(S)-methylpentyl]-N-1 -
naphthylmethyl-glycyl-methionine methyl ester bis
trifluoroacetate (19) and N-[2(S)-(1-(4-Nitrophenyl-methyl)-
1 H-imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
naphthylmethyl-glycyl-methionine methyl ester bis
trifluoroacetate (20)
To a solution of 1-(4-nitrophenylmethyl)-lH-imidazol-4-
ylacetic acid hydrochloride (17) and 1-(4-nitrophenylmethyl)-lH-
imidazol-5-ylacetic acid hydrochloride (18, 3:1 mixture, 153 mg, 0.392
mrnol), N-[2(S)-amino-3(S)-methylpentyl]-N-naphthylmethyl-glycyl-
methionine methyl ester bis hydrochloride (10, 209 mg, 0.392 mmol) and
3-hydroxy-1,2,3-benzotriazin-4(3H)-one (HOOBT, 64 mg, 0.39 mmol) in
methylene chloride (10 ml) was added 1-(3-dimethylaminopropyl)-3-
ethylcarbodiimide hydrochloride (EDC, 75.2 mg, 0.392 mmol) and
triethyl~mine (219 ,ul, 1.57 mmol) and the mixture stirred overnight at
room temperature. After this time, sat. aq. sodium bicarbonate (10 ml)
was added and the mixture was extracted with methylene chloride. The
combined extracts were washed with sat. aq. sodium bicarbonate (10 ml)
and the solvent evaporated in vacuo. The regioisomers were separated
by preparative HPLC (chromatography method A ) to give after
lyophili7~tion 19 and 20.
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19:
lH NMR (CD30D, 400 MHz) ~ 8.96 (lH, s), 8.17 (lH, m), 8.Z3 (2H, d,
J=8.7 Hz), 7.92 (2H, d, J=8.9 Hz), 7.61 (lH, d, J=6.9 Hz), 7.56 (2H, d,
J=8.9 Hz), 7.50 (2H, m), 7.44 (2H, m), 5.52 (2H, s), 4.70 (lH, d, J=9.4
Hz),4.49(1H,d,J=11.9Hz),4.38(1H,dd,J=4.7and8.9Hz),4.13 (lH,
m), 3.67 (3H, s), 3.65 (4H, m), 3.30 (lH, m), 3.06 (lH, m), 2.31 (lH, m),
2.23 (lH, m), 1.97 (3H, s), 1.94 (lH, m), 1.71 (lH, m), 1.57 (lH, m),
1.42 (lH, m), 1.17 (lH, m), 0.90 (3H, d, J=6.9 Hz) and 0.87 (3H, t, J=7.4
Hz) ppm.
o Anal. Calcd for C37H46N606S-2.40 TFA-0.25 H2O: C, 51.18; H,5.02;
N, 8.57. Found: C, 51.17; H, 5.03; N, 8.80.
FAB MS calcd for C37H47N606S 703 (MH+), found 703.
20:
H NMR (CD30D, 400 MHz) ~ 8.91 (lH, s), 8.26 (lH, d, J=12.8 Hz),
8.21 (2H, d, J=10.7 Hz), 7.91 (2H, m), 7.65-7.36 (7H, m), 5.51 (2H, s),
4.72-3.99 (4H, m), 3.66 (3H, s), 3.66-3.24 (4H, m), 3.20-2.85 (2H, m),
2.29 (lH, m), 2.20 (lH, m), 1.96 (3H, s), 1.91 (lH, br s), 1.70 (lH, d,
J=16 Hz), 1.56 (lH, m), 1.38 (lH, m), 1.13 (lH, m) and 0.88 (6H, m)
ppm.
FAB HRMS exact mass calcd for C37H47N606S 703.32778 (MH+),
found 703.32852.
Step D: Preparation of N-[2(S)-(1 -(4-Nitrophenylmethyl)-lH-
imidazol-4-ylacetyl)amino-3(S)-methylpentyl] -N- 1 -
naphthylmethvl-~lycyl-methionine bis trifluoroacetate (21)
To a solution of N-[2(S)-(1-(4-nitrophenylmethyl) lH-
imidazol-4-ylacetyl)amino-3(S)-methylpentyl]-N- l -naphthylmethyl-
glycyl-methionine methyl ester bis trifluoroacetate (19, 21 mg, 0.023
mmol) in methanol (1 ml ) at room temperature was added l.ON lithium
hydroxide (135 ~l, 0.135 mmol). This solution was stirred for 4 h and
treated with trifluoroacetic acid (100 ,ul). This mixture was puri~led by
preparative HPLC using chromatography method A to give 21.
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lH NMR (CD30D, 400 MHz) ~ 8.86 (lH, s), 8.23 (2H, d, J= 8.8Hz),
8.22 (lH, m), 7.90 (2H, dd, J=7.3 Hz), 7.55 (2H, d, J=8.4 Hz), 7.44-7.2~s
(5H, m), 5.50 (2H, s), 4.53 (lH, m), 4.35 (2H, m), 4.12 (lH, m), 3.79-
3.25 (4H, m), 3.26-2.86 (2H, m), 2.27 (lH, m), 2.18 (lH, m), 1.96 (3H,
s), 1.9 (lH, m), 1.67 (lH, m), 1.57 (lH, m), 1.42 (lH, m), 1.15 (lH, m),
0.90 (3H, d, J=6.9 Hz) and 0.86 (3H, t, J=7.3 Hz) ppm.
FAB HRMS exact mass calcd for C36H45N606S 689.31213 (MH+),
found 689.31262.
Step E: Preparation of N-[2(S)-(1-(4-Nitrophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
naphthylmethyl-~lycyl-methionine bis trifluoroacetate (22)
To a solution of N-[2(S)-N'-(1-(4-nitrophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl] -N- 1 -naphthylmethyl-
glycyl-methionine methyl ester bis trifluoroacetate (20, 29 mg, 0.031
mmol) in methanol (1 ml ) was added l.ON lithium hydroxide (187 ,ul,
0.187 mmol ). This solution was stirred for 4 h and treated with
trifluoroacetic acid (100,ul). This mixture was purified by preparative
HPLC using chromatography method A to give 22.
lH NMR (CD30D, 400 MHz) ~ 8.89 (lH, s), 8.25 (lH? m), 8.21 (2H, d,
J= 9.0Hz), 7.89 (2H, m), 7.64-7.34 (7H, m), 5.52 (2H, s), 4.59-3.88 (4H,
m), 3.77-3.38 (4H, m), 3.18-2.75 (2H, m), 2.27 (lH, m), 2.18 (lH, m),
1.96 (3H, s), 1.9 (lH, m), 1.67 (lH, m), 1.57 (lH, m), 1.42 (lH, m), 1.15
(lH, m), 0.89 (6H, m) ppm.
FAB HRMS exact mass calcd for C36H4sN606S 689.31213 (MH~),
found 689.31135.
EXAMPLE 3
Regioselective preparation of N-[2(S)-(1 -(4-Nitrophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
~lycvl-methionine methyl ester bis trifluoroacetate (20)
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Step A: Preparation of 1-(Triphenylmethyl)-lH-imidazol-4-ylacetic
acid methyl ester (23)
To a suspension of lH-imidazole-4-acetic acid methyl ester
hydrochloride (1,7.48,42.4 mmol) in methylene chloride (200 ml) was
added triethyl~mine (17.7 ml,127 mmol) and triphenylmethyl bromide
(16.4 g,50.8 mmol) and stirred for 72 h. After this time, reaction mixture
was washed with sat. aq. sodium bicarbonate (100 ml) and water (100
ml). The organic layer was evaporated in vacuo and purified by flash
chromatography (30-100% ethyl acetate/hexanes gradient elution) to
provide 23 as a white solid.
lH NMR (CDCl3,400 MHz) ~ 7.35 (lH, s),7.31 (9H, m),7.22 (6H, m),
6.76 (lH, s),3.68 (3H, s) and 3.60 (2H, s) ppm.
Step B: Preparation of 1 -(4-Nitrophenylmethyl)-lH-imidazol-5-
ylacetic acid methyl ester (16)
To a solution of l-(triphenylmethyl)-lH-imidazol-4-ylacetic
acid methyl ester (23, 274 mg, 0.736 mmol) in acetonitrile (10 ml) was
added 4-nitrobenzylbromide (159 mg, 0.736 mmol) and heated to 55C
for 16 h. After this time, the reaction was cooled to room temperature,
treated with ethyl acetate (20 ml) and the resulting precipitate was
filtered. The filtrate was concentrated to dryness in vacuo and the residue
was redissolved in acetonitrile (4 ml) and heated to 65C for 3 h. After
this time, the reaction mixture was evaporated to dryness and combined
with initial precipitate. This residue was dissolved in methanol (5 ml )
and heated to reflux for 30 min. The resulting solution was evaporated in
vacuo and the residue was purified by flash chromatography (2-5%
methanol/methylene chloride gradient elution ) to provide 16.
1H NMR (CDC13,400 MHz) ~ 8.20 (2H, d, J=8.8 Hz),7.53 (lH, s),7.19
(2H, d, J=8.8 Hz),7.03 (lH, s),5.28 (2H, s), 3.61 (3H, s) and 3.44 (2H, s)
ppm.
Step C: Preparation of 1 -(4-Nitrophenylmethyl)-1 H-imidazol-5-
ylacetic acid hydrochloride (18)
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1-(4-Nitrophenylmethyl)-lH-imidazol-5-ylacetic acid
methyl ester (0.115 g, 0.42 mmol ) was dissolved in l .ON hydrochloric
acid (10 ml ) and heated at 55C for 3 h. The solution was evaporated in
vacuo to give 18 as a white solid.
lH NMR (CD30D, 400 MHz) ~ 9.06 (lH, s), 8.27 (2H, d, J=8.8 Hz),
7.61 (lH, s), 7.55 (2H, d, J=8.8 Hz), 5.63 (2H, s) and 3.81 (2H, s) ppm.
Step D: Preparation of N-[2(S)-(1-(4-Nitrophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
o naphthylmethyl-glycyl-methionine methyl ester bis
trifluoroacetate (20)
Following the procedure described in Example 2, Step C,
but using the 1-(4-nitrophenylmethyl)-lH-imidazol-5-ylacetic acid
hydrochloride, prepared as described in Step C provided the title
compound.
EXAMPLE 4
Preparation of N-[2(S)-(1 -(2-Naphthylmethyl)- 1 H-imidazol-5 -
ylacetyl)amino-3(S)-methylpentyl] -N- 1 -naphthylmethyl-glycyl-
methionine bis trifluoroacetate
Step A: Preparation of N-[2(S)-(1 -(2-Naphthylmethyl)- lH-imidazol-
5-ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -naphthylmethyl -
glycyl-methionine methyl ester bis trifluoroacetate
Following the procedure described in Example 3, ~teps B-D,
but using 2-(bromomethyl)naphthlene in place of 4-nitrobenzylbromide
provided the title compound.
lH NMR (CD30D, 400 MHz) ~ 8.89 (lH, s), 8.29 (lH, d, J=9 Hz), 7.92
(4H,m),7.83 (lH,d,J=9Hz),7.68(1H,s),7.58-7.42(7H,m),7.33 (lH,
d, J=9 Hz), 5.54 (2H, s),4.90-4.50 (2H, m), 4.38 (lH, m), 4.05 (lH, m),
3.93-3.32 (5H, m), 3.65 (3~I, s), 3.12 (lH, m), 2.24 (2H, m), 1.93 (3H, s),
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1.87 (lH, br s), 1.72 (lH, br s), 1.52 (lH, br s), 1.38 (lH, br ~), 1.13 (lH,
br s) and 0.87 (6H, m) ppm.
Anal. Calcd for C41H49N504S-3.20 TFA-0.75 H20: C, 52.41; H, 4.98;
N, 6.45. Found: C, 52.40; H, 4.96; N, 6.63.
FAB HRMS exact mass calcd for C41H50N5O4S 708.358352 (MH+),
found 708.357618.
StepB: Preparationof N-[2(S)-(1-(2-Naphthylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
o naphthylmethvl-~lycvl-methionine bis trifluoroacetate
Following the procedure described in Example 2, Step E, but
using the methyl ester prepared as described in Step A provided the title
compound.
H NMR (CD30D, 400 MHz) ~ 8.88 (lH, s), 8.28 (lH, d, Y=9 Hz),
7.96-7.78 (SH, m), 7.67 (lH, s), 7.57-7.41 (7H, m), 7.32 (lH, d, J=9 Hz),
5.55(2H,s),4.81(1H,m),4.56(1H,m),4.37(iH,m),4.06~1H,m),
3.89-3.50(4H,m),3.42(1H,m),3.10(1H,m),2.28(1H,m),2.19(1H,
m), 2.03-1.86 (lH, m), 1.93 (3H, s), 1.90 (lH, m), 1.71 (lH, m), 1.52
(lH, m), 1.37 (lH, m) and 0.87 (6H, m) ppm.
20 Anal. Calcd for C40H47N504S-2.95 TFA-0.5 H20: C, 53.05; H, 4.94;
N, 6.74. ~ound: C, 53.03; H, 4.95, N, 7.10.
FAB HRMS exact mass calcd for C40H48N5o4s 694.342702 (MH+),
found 694.342837.
2s EXAMPLE 5
Preparation of N-~2(S)-(1-(1-Naphthylmethyl)-lH-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-
methionine bis trifluoroacetate
Step A: Preparation of N-[2(S)-(1-(l-Naphthylmethyl)-lH-imidazol-
5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
~lycyl-methionine methyl ester bis trifluoroacetate
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Following the procedure described in Example 3, Steps A-D,
but using l-(bromome~yl)naphthlene in place of 4-nitrobenzylbromide
r provided ~e ~le compound.
lH NMR (CD30D, 400 MHz) ~ 8.42 (lH, s) 8.31 (lH, d, J=8.9 Hz),
8.04-7.80 (SH, m), 7.69 (lH, m), 7.59-7.39 (7H, m), 7.20 (lH, d, J=8.2
Hz), 5.80 (2H, s),5.0-4.5 (2H, m), 4.26 (lH, m), 4.13 (lH, m), 4.0-3.6
(4H, m), 3.64 (3H, s), 3.49 (lH, m), 3.18 (lH, m), 2.17 (2H, m), 1.91
(3H, s), 1.86 (lH, m), 1.67 (lH, m), 1.55 (lH, m), 1.41 (lH, m), 1.16
(lH, br s), and 0.88 (6H, m) ppm.
Anal. Calcd for C41H49N5O4S-3.10 TFA-0.55 H2O: C, 52.92; H, 5.01;
N, 6.54. Found: C, 52.90; H, 4.99, N, 6.59.
FAB HRMS exact mass calcd for C41H50N5O4S 708.358352 (MH+),
found 708.357618.
Step B: Preparation of N-[2(S)-(l-(l-Naphthylmethyl)-lH-imidazol-
5-ylacetyl)amino-3(S)-methylpentyl3-N- I -naphthylmethyl-
~lycyl-methionine bis trifluoroacetate
Following the procedure described in Example 2, Step E, but
using the methyl ester prepared as described in Step A provided the title
compound.
lH NMR (CD30D, 400 MHz) ~ 8.41 (lH, s), 8.19 (lH, d, J=7.7 Hz),
7.99 (2H, m), 7.87 (3H, m), 7.64 (lH, m), 7.56 (lH, t, J=7 Hz), 7.46 (6H,
m), 7.16 (lH, d, J=8 Hz),5.79 (2H, s), 5.04-4.71 (lH, m), 4.61-4.38 (lH,
m), 4.38-4.21 (lH, m), 4.14 (lH, m), 3.97-3.51 (4H, m), 3.51-3.21 (lH,
2s m), 3.21-2.85 (lH, m), 2.21 (lH, m), 2.13 (lH, m), 1.98 (lH, m), 1.91
(3H, s), 1.66 (lH, m), 1.56 (lH, m), 1.40 (lH, m), 1.15 (lH, m), and 0.87
(6H, m) ppm.
Anal. Calcd for C40H47N504S-2.70 TFA-0.5 H20: C, 53.95; H, 5.06;
N, 6.93. Found: C, 53.97; H, 5.06; N, 7.10.
FAB HRMS exact mass calcd for C40H48N5O4S 694.342702 (MH+),
found 694.342837.
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EXAMPLE 6
Preparationof N-[2(S)-(l-Farnesyl-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl]-N-1-naphthylmethyl-glycyl-methionine bis
trifluoroacetate
Step A: Preparation of l-Farnesyl-lH-imidazol-5-ylacetic acid
metkyl ester
To a solution of l-(triphenylmethyl)-lH-imidazol-4-ylacetic
o acid methyl ester (200 mg, 0.523 mmol) in acetonitrile (5 ml) was added
trans, trans-farnesyl bromide (156 ~1, 0.575 mmol) and heated at 55C
for 16 h. After this time, the reaction was heated at 80C for 3 h and then
the reaction mixture was evaporated in vacuo. The residue was dissolved
in methanol (S ml ) and heated to reflux for 30 min and then evaporated
in vacuo. The residue was purified by flash chromatography ~2-4%
methanol/methylene chloride gradient elution) to provide the title
compound.
lH NMR (CDC13, 400 MHz) ~ 7.50 (lH, s), 6.92 (lH, s), 5.24 (lH, t,
J=5.9 Hz), 5.09 (2H, m), 4.49 (2H, d, J=6.9 Hz), 3.69 (3H, s), 3.60 (2H,
s), 1-91-2.15 (8H, m), 1.72 (3H, s), 1.65 (3H, s), 1.59 (3H, s) and 1.57
(3H, s) ppm.
Step B: Preparation of N-[2(S)-(1-(1-Farnesyl)-lH-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthy]methyl-
~lvcyl-methionine methyl ester bis trifluoroacetate
Following the procedure described in Example 3, Steps C-D,
but using 1-farnesyl-lH-imidazol-5-ylacetic acid methyl ester described
in Step A in place of 1-(4-nitrophenylmethyl)-lH-imidazol-5-ylacetic
acid methyl ester provided the title compound.
lH NMR ((~D30D, 400 MHz) ~ 8.70 (lH, s), 8.26 (lH, m), 7.91 (2H,
m), 7.52 (3H, m), 7.48 (lH, m), 7.37 (lH, s), 5.40 (lH, m~, 5.08 (2H, m),
4.94-4.72 (3H, m), 4.71 (lH, m), 4.40 (lH, m), 4.13 (lH, m), 3.95-2.80
(6H, m), 3.68 ~3H, s), 2.27 (lH, m), 2.21 (lH, m), 2.09 (8H, m), 1.97
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(3H, s), 1.92 (2H, m), 1.72 (3H, s), 1.65 (lH, m), 1.65 (3H, s), 1.60 (3H,
s), 1.58 (3H, s), 1.42 (lH, m), 1.18 (lH, m) and 0.90 (6H, m) ppm.
FAB HRMS exact mass Calcd for C45H66N5O4S 772.483553 (MH+),
- found 772.481709.
Step C: Preparation of N-[2(S)-[l-(l-Farnesyl)-lH-imidazol-5-
ylacetyl] amino-3 (S)-methylpentyl] -N- 1 -naphthylmethyl -
glycyl-methionine bis trifluoroacetate
Following the procedure described in Example 2, Step E, but
o using the methyl ester prepared as described in Step B provided the title
compound.
lH NMR (CD30D, 400 MHz) o 8.68 (lH, s), 8.18 (lH, m), 7.90 (2H,
m), 7.52 (3H, m), 7.44 (lH, t, J=7.5 Hz), 7.37 (lH, s), 5.29 (lH, br t, J=7
Hz), 5.08 (2H, m), 4.95-4.64 (lH, m), 4.73 (2H, m), 4.37 (2H, m), 4.12
(lH, m), 3.71 (2H, m), 3.47 (2H, m), 3.11 (lH, m), 2.95 (lH, m), 2.27
(lH, m), 2.23-2.01 (9H, m), 2.01-1.89 (lH, m), 1.97 (3H, s), 1.77-1.54
(2H, m), 1.71 (3H, s), 1.65 (3H, s), 1.60 (3H, s), 1.58 (3H, s), 1.42 (lH,
m), 1.16 (lH, m), 0.91 (3H, t, J=7 Hz) and 0.87 (3H, d, J=7.5 Hz) ppm.
FAB HRMS exact mass calcd for C44H64N5O4S 758.467903 (MH,
found 758.467591.
EXAMPLE 7
Preparationof N-[2(S)-(l-Geranyl-lH-imidazol-5-ylacetyl)amino-3(S)-
methylpentyl] -N- 1 -naphthylmethyl-glycyl-methionine bis
trifluoroacetate
Step A: Preparation of N-[2(S)-(l-Geranyl-lH-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N-l-naphthylmethyl-
glycyl-methionine methyl ester bis trifluoroacetate
Following the procedure described in Fx~mple 6, Steps A-B,
but using trans-geranyl bromide in place of farnesyl bromide provided the
title compound.
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lH NMR (CD30D, 400 MHz) ~ 8.67 (lH, s), 8.27 (lH, m), 7.92 (2H,
m), 7.57 (lH, m), 7.53 (2H, m), 7.46 (IH, dd, J=9 Hz), 7.36 (IH, s), 5.29
(lH,t,J=6Hz),5.08 (lH,t,J=6Hz),4.71 (lH,m),4.71-4.12(1H,m),
4.38(1H,m),4.12(1H,m),3.80-3.33(4H,m),3.68(3H,s),3.14(1H,
m),2.96(1H,m~,2.29(1H,m),2.21 (lH,m),2.12(4H,m),2.Il (lH,
m), 1.97 (3H, s), 1.97 (lH, m), 1.70 (3H, s), 1.68 (3H, s), 1.65 (lH, m),
1.60 (3H, s), 1.41 (lH, m), 1.15 (lH, m), 0.91 (3H, d, J=7 Hz) and 0.88
(3H, t, J=7.5 Hz) ppm.
Anal. Calcd for C40H57N504S-1.80 TFA-0.25 H20: C, 57.31; H, 6.54,
N, 7.66. Found: C, 57.2P~; H, 6.54; N, 7.90.
FAB HRMS exact mass calcd for C40HsgNsO4S 704.420953 (MH~),
found 704.420223.
Step B: Preparation of N-~2(S)-(I-Geranyl-lH-imidazol-5-
ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -naphthylme~yl -
~lycyl-methionine bis trifluoroacetate
Following the procedure described in Example 2, Step E, but
using the methyl ester prepared as described in Step A provided the title
compound.
20 lH NMR (CD30D, 400 MHz) o 8.67 (lH, s), 8.27 (lH, m), 7.92 (2H,
m), 7.59 (lH, m), 7.52 (2H, m), 7.46 (lH, t, J-7.8 Hz), 7.38 (lH, s), 5.28
(IH, t, J=11.2 Hz), 5.04 (lH, m), 4.96-4.54 (lH, m), 4.72 (2H, s), 4.54-
4.31 (lH, m), 4.39 (lH, m), 4.13 (lH, m), 3.82-3.31 (4H, m), 3.68 (2H,
m), 3.31-2.79 (2H, m), 2.30 (lH, m), 2.12 (SH, m), 1.97 (3H, s), 1.97
2s (lH, m), 1.73 (lH, m), 1.71 (3H, s), 1.70 (3H, s), 1.60 (3H, s), 1.44 (lH,
m), 1.18 (lH, m~ and 0.92 (3H, d, J=6.8 Hz), and 0.90 (3H, t, J=7.5 Hz)
ppm.
FAB HRMS exact mass calcd for C39H56N504S 690.405303 (MH+),
found 690.405157.
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EXAMPLE 8
Preparation of N-[2(S)-(1-(4-Pyridylmethyl)-lH-imidazol-4-
ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -naphthylmethyl-glycyl-
methionine tris trifluoroacetate (28) and N-[2(S)-(1-(4-Pyridylmethyl)-
1 H-imidazol-5-ylacetyl)amino-(3S)-methylpentyl]-N- 1 -naphthylmethyl-
glycvl-methionine tris trifluoroacetate (29)
Step A: Preparation of 1-(4-Pyridylmethyl)-lH-imidazol-4-ylacetic
acid methyl ester (24) and 1-(4-Pyridylmethyl)-lH-imidazol-
5-ylacetic acid methyl e~ster (25) (3: 1 mixture)
To a solution of sodium hydride (60% in mineral oil, 99 mg,
2.5 mmol) in dimethylform~mide (2 ml) cooled at 0C over ice bath was
added, via c~nn~ , a solution of lH-imidazole-4-acetic acid methyl ester
hydrochloride (1, 115 mg, 0.707 mmol) in dimethylformamide (2 ml).
The suspension was stirred at 0C for 15 min. This suspension was
added to a solution prepared by adding 4-picolyl chloride hydrochloride
(185 mg, 0.707 mmol) to sodium hydride (60% in mineral oil, 45.2 mg,
1.13 mmol) in dimethylform~mide (2 ml ) at 0C. After the addition was
complete, the lni~lul~ was stirred at 0C for 15 min and then at room
temperature for 1.5 h. After this time, the mixture was quenched with
sat. aq. sodium bicarbonate (50 ml) and extracted with methylene
chloride (2 X 50 ml). The combined organic extracts were washed with
brine (50 ml), dried(MgSO4), filtered and the solvent evaporated in
vacuo. The residue was purified by flash chromatography( 3-7%
methanol/methylene chloride gradient elution) to give a 3: 1 mixture of 24
and 25.
lH NMR (CDCL3, 400MHz) ~ 8.57 (1.5H, d, J=5 Hz), 8.56 (0.5H, d,
J=7 Hz), 7.51 (0.25H, s), 7.46 (0.75H, s), 7.01 (0.25H, s), 6.99 (1.5H, d,
J=5 Hz), 6.90 (0.5H, d, J=7 Hz), 6.86 (0.75H, s), 5.17 (0.5H, s), 5.08
(1.5H, s), 3.69 (2.25H, s), 3.64 (1.5H, s), 3.58 (0.75H, s) and 3.43 (0.5H,
s) ppm.
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Step B: Preparation of N-[2(S)-(1-(4-Pyridylmethyl)-lH-imidazol-4-
ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -naphthylmethyl-
glycyl-methionine methyl ester tris trifluoroacetate (26) and
N-[2(S)-(1 -(4-Pyridylmethyl)- 1 H-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N-l-naphthylmethyl-
glvcyl-methionine methyl ester tris trifluoroacetate (27)
Following the procedure described in Example 2, Steps B-C,
but using the mix~lre of pyridylmethylimidazolylacetic acid from Step A
provided the title compounds after preparative HPLC.
26: lH NMR (CD30D, 400 MHz) o 8.99 (lH, s), 8.65 (2H, d, J=4.9
Hz), 8.28 (lH, d, J=9.4 Hz), 7.91 (2H, m), 7.69 (lH, d, J=6.5 Hz), 7.61-
7.44 (6H, m), 5.59 (2H, s), 4.90 (lH, m), 4.68 (lH, d, J=13.4 Hz), 4.42
(lH, m), 4.16 (lH, m), 3.90 (lH, d, J=15.6 Hz), 3.82 (lH, d, J=15.6 Hz),
3.75-3.55 (2H, m), 3.69 (3H, s), 3.50 (lH, d, J=13.1 Hz), 3.20 (lH, m),
2.37 (lH, m), 2.29 (lH, m), 1.99 (3H, s), 1.96 (lH, m), 1.77 (lH, m),
1.58 (lH, m), 1.23 (lH, m), 1.19 (lH, m) and 0.91( 6H, m) ppm.
Anal. Calcd for C36H46N604S-4.95 TFA-2.2 H20: C, 43.65; H, 4.42;
N, 6.65. Found: C, 43.65; H, 4.16; N, 6.68.
20 FAB HRMS exact mass calcd for C36H47N604S 659.337951 (MH+),
found 659.336943
27: lH NMR (CD30D, 400 MHz) ~ 9.01 (lH, s), 8.63 (2H, m), 8.28
(lH, m), 7.98 (2H, m), 7.70 (lH, d, J=6.0 Hz), 7.52 (4H, m), 7.41 (2H, d,
25 J=6.2 Hz), 5.62 (2H, s), 4.94 (lH, m), 4.72 (lH, m), 4.42 (lH, m), 4.07
(lH, m), 3.89 (2H, m), 3.68 (lH,m), 3.69 (3H, s), 3.55 (2H, m), 3.24 (lH,
m),2.39(1H,m),2.31 (lH,m),2.00(3H,s), 1.98(1H,m), 1.79(1H,m),
1.58 (lH, m), 1.42 (lH, m), 1.18 (lH, m) and 0.91 (6H, m) ppm.
FAB HRMS exact mass calcd for C36H47N604S 659.337951 (MH+),
30 found 659.336826.
Step C: Preparation of N-[2(S)-(1-(4-Pyridylmethyl)-lH-imidazol-4-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
glvcyl-methionine methyl ester tris trifluoroacetate (28)
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Following the procedure described in Example 2, Step D,
but using the methyl ester 26 prepared as described in Step B provided
the title compound.
lH NMR (CD30D, 400 MHz) ~ 8.96 (lH, s), 8.55 (2H, d, J=5.2Hz),
8.21 (lH, d, J=7.2 Hz), 7.97 (2H, m), 7.69 (lH, d, J=7.2 Hz), 7.60-7.40
(6H, m), 5.58 (2H, s), 4.91 (lH, d, J=13.2 Hz), 4.69 (lH, d, J=13.2 Hz),
4.38 (lH, dd, J=4.6 and 8.8 Hz), 4.15 (lH, m), 3.89 (lH, d, J=16.1 Hz),
3.81 (lH, d, J=16.1 Hz), 3.71 (lH, d, J=17 Hz), 3.62 (lH, d, J=17 Hz),
3.50 (lH, dd, J=3.4 and 12 Hz), 3.21 (lH, m), 2.38 (lH, m), 2.27 (lH,
m), 1.99 (lH, m), 1.99 (3H, s), 1.77 (lH, m), 1.58 (lH, m), 1.43 (lH, m),
1.16 (lH, m), and 0.88 (6H, m) ppm.
FAB HRMS exact mass calcd for C35H45N6O4S 645.322301 (MH+),
found 645.323649.
5 Step D: Preparation of N-[2(S)--(1-(4-Pyridylmethyl)-lH-imidazol-
5-ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -naphthylmethyl-
~lycyl-methionine tris trifluoroacetate (29)
Following the procedure described in Example 2, Step E, but
using the methyl ester 27 prepared as described in Step B provided the0 title compound.
lH NMR (CD30D, 400 MHz) ~ 8.97 (lH, s), 8.58 (2H, s), 8.27 (lH, m),
7.95 (2H, m), 7.64 (lH, m), 7.50 (4H, m), 7.31 (2H, d, J=4.4 Hz), 5.57
(2H, s), 4.63 (2H, m), 4.38 (lH, m), 4.09 (lH, m), 3.78 (2H, m), 3.60
25 (2H,m),3.42(1H,m),3.15(1H,m),2.36(1H,m),2.15(1H,m),2.01
(lH, m), 1.98 (3H, s), 1.76 (lH, m), 1.55 (lH, m), 1.41 (lH, m), 1.15
(lH, m) and 0.88 (6H, m) ppm.
FAB HRMS exact mass calcd for C35H45N604 645.322301 (MH+),
found 645.321321.
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EXAMPLE 9
Preparation of N-[2(S)-(1 -(4-Cyanophenylmethyl)- 1 H-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-
5 methionine bis trifluoroacetate
Step A: Preparation of N-[2(S)-(1 -(4-Cyanophenylmethyl~-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
naphthylmethyl-glycyl-methionine methylesterbis
trifluoroacetate
Following the procedure described in Example 3, Steps B-D,
but using a-bromo-p-tolunitrile in place of 4-nitrobenzylbromide
provided the title compound.
H NMR (CD30D, 400 MHz) ~ 8.92 (lH, s), 8.31 (lH, m), 8.01 (lH, d,
J=8 Hz), 7.96 (lH, m), 7.75 (2H, d, J=8 Hz), 7.62 (lH, s), 7.58-7.48 (3H,
m), 7.45 (lH, m), 7.41 (2H, d, J=8 Hz), 5.51 (2H, s~, 4.97 (lH, m), 4.76
(lH, m), 4.41 (lH, m), 4.10 (lH, m) 3.92 (2H, m), 3.75-3.47 (3H, m),
3.69 (3H, s), 3.25 (lH, m), 2.37 (lH, m), 2.30 (lH, m), 2.00 (3H, s), 1.97
(lH,m), 1.79 (lH, m), 1.58 (lH, m), 1.43 (lH, m), 1.19 (lH, m) and 0.91
20 (6H, m~ ppm.
Anal. Calcd for C38H46N6O4S-2.40 TFA-1.90 H2O: C, 51.89; H, 5.31;
N, 8.48. Found: C, 51.88; H, 5.29; N, 8.72.
FAB HRMS exact mass calcd for C3gH47N6O4S 683.337951 (MH+),
found 683.338437.
Step B: Preparation of N-[2(S)-(1-(4-Cyanophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
naphthylmethvl-glycvl-methionine bis trifluoroacelate
To a solution of N-[2(S)-(1-(4-cyanophenylmethyl)-lH-
30 imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
glycyl-methionine methyl ester bis trifluoroacetate (25.6 mg, 0.028
mmol) in methanol (1 ml ) was added l.ON sodium hydroxide (280 ,ul,
0.280 mmol) and stirred for 2 h. After this time, the mixture was treated
with trifluoroacetic acid (to pH <3) and purified by preparative HPLC
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(chromatography method A) to give after lyophili~tion, the title
compound.
lH NMR (CD30D, 400 MHz) ~ 8.87 (lH, s), 8.27 (lH, d, J=9.2 Hz),
7.90 (2H, m), 7.73 (2H, d, J=8 Hz), 7.60 (lH, s), 7.46 (4H, m), 7.36 (2H,
d, J=8 Hz), 5.48 (2H, s), 4.95-4.28 (2H, m), 4.36 (lH, m), 4.09 (lH, m),
3.59 (4H, m), 3.51-2.73 (2H, m), 2.29 (lH, m), 2.19 (lH, m), 2.03-1.85
(lH, m), 1.97 (3H, s), 1.70 (lH, m), 1.56 (lH, m), 1.39 (lH, m), 1.14
(lH, m) and 0.89 (6H, m) ppm.
Anal. Calcd for C37H44N604S-2.45 TFA-1.3 H20: C, 51.80; H, 5.09;
N, 8.65. Found: C, 51.78; H, 5.07; N, 8.95.
FAB HRMS exact mass Calcd for C37H44N6O4S 669.322301 (MH+),
found 669.323148.
EXAMPLE 10
Preparation of N-[2(S)-(1 -(4-Methoxyphenylmethyl)- 1 H-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-
methionine bis trifluoroacetate
Step A: Preparation of N-[2(S)-(1-(4-Methoxyphenylmethyl)-lH-
imidazol-5-yl)acetyl)amino-3(S)-methylpentyl]-N-l -
naphthylmethyl-glycyl-methionine methyl ester bis
trifluoroacetate
Following the procedure described in Example 3, Steps B-D,
but using 4-methoxybenzyl chloride in place of 4-nitrobenzylbromide
provided the title compound.
lH NMR (CD30D, 400 MHz) ~ 8.70 (lH, s), 8.27 (lH, m), 7.92 (2H,
m), 7.70-7.35 (SH, m),7.18 (2H, d, J=8.5 Hz), 6.92 (2H, d, J=8.5 Hz),
5.27 (2H, s), 4.60-4.00 (4H, m), 3.79 (3H, s), 3.67 (3H, s), 3.61 (4H, m),
3.40-2.75 (2H, m), 2.28 (lH, m), 2.19 (lH, m), 1.96 (3H, s), 1.91 (lH,
m), 1.70 (lH, m), 1.60 (lH, m), 1.43 (lH, m), 1.18 (lH, m) and 0.91 (6H,
m) ppm-
Anal. Calcd for C38H49NsO5S-1.75 TFA-1.75 H2O: C, 54.45; H, 5.98;
N, 7.67. Found: C, 54.44; H, 5.95; N, 7.85.
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FAB HRMS exact mass calcd for C3gHsoN5O5S 688.353267 (MH+),
found 688.352186.
Step B: Preparation of N-[2(S)-(1-(4-Methoxyphenylmethyl)-lH- imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N-1-
naphthylmethyl-~lycyl-methionine bis trifluoroacetate
Following the procedure described in Example 9, Step B,
but sub~ g the methyl ester from Step A provided the title
compound.
10 lH NMR (CD30D, 400 MHz) ~ 8.70 (lH, s), 8.27 (lH, m), 7.92 (2H,
m), 7.63 (lH, s), 7.56-7.35 (4H, m), 7.18(2H, d, J=8.6 Hz), 6.93 (2H, d,
J=8.6 Hz), 5.27 (2H, s), 4.93-4.29 (2H, m), 4.36 (lH, m), 4.12 (lLH, m),
3.79 (3H, s), 3.63 (4H, m), 3.07 (2H, m), 2.28 ~lH~ m), 2.19 (lH, m),
2.02-1.88 (lH, m), 1.95 (3H, s), 1.70 (lH, m), 1.60 (lH, m), 1.43 (lH,
15 m), 1.18 (lH, m) and 0.91 (6H, m) ppm.
FAB HRMS exact mass calcd for C37H4gN5O5S 674.337617 (MH+),
found 674.338053.
EXAMPLE 11
Preparation of N-[2(S)-(1-(4-Quinolinylmethyl)-lH-imidazol-S-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-
methionine bis trifluoroacetate
5 Step A: Preparation of N-[2(S)-(1-(4-Quinolinylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N-l -
naphthylmethyl-glycyl-methionine methyl ester bis
trifluoroacetate
Following the procedure described in Example 3, Steps B-D,
30 but using 4-(bromomethyl)quinoline hydrochloride in place of 4-
nitrobenzylbromide provided the title compound.
lH NMR (CD30D, 400 MHz) ~ 8.88 (lH, s), 8.83 (lH, d, J=4.8 Hz),
8.28 (lH, m), 8.15 (lH, d, J=8.6 Hz), 7.99-7.85 (4H, m), 7.67 (2H, m),
7.57 (lH, s), 7.48 (3H, m), 6.96 (lH, m), 6.02 (2H, s), 4.90 (lH, m), 4.62
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(lH, m~, 4.18 (lH, m), 4.07 (lH, m), 3.94-3.50 (4H, m), 3.64 (3H, s),
3.45 (lH, m), 3.13 (lH, m), 2.2~ (lH, m), 2.21 (lH, m), 1.95 (3H, s),
1.87 (lH, m), 1.69 (lH, m), 1.48 (lH, m), 1.35 (lH, m), 1.11 (lH, m) and
0.84 (6H, m) ppm.
FAB HRMS exact mass calcd for C40H49N6O4S 709.353601 (MH+),
found 709.353711.
Step B: Preparation of N-[2(S)-(1-(4-Quinolinylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl] -N- 1 -
naphthylmethyl-glycvl-methionine bis trifluoroacetate
Following the procedure described in Example 9, Step B,
but substituting the methyl ester from Step A provided the title
compound.
H NMR (CD30D, 400 MHz) ~ 8.87 (lH, s), 8.82 (lH, d, J=5 Hz), 8.28
(lH, m), 8.15 (lH, d, J=8.6 Hz),8.06-7.82 (4H, m), 7.67 (2H, m), 7.58
(lH, s), 7.48 (3H, s), 6.96 (lH, m), 6.03 (2H, s), 4.93-4.57 (2H, m), 4.22
(lH, m), 4.08 (lH, m), 3.72 (4H, m), 3.47 (lH, m), 3.13 (lH, m), 2.28
(lH, m), 2.21 (lH, m), 1,95 (3H, s), 1.87 (lH, m), 1.70 (lH, m), 1.4
(lH, m), 1.35 (lH, m), 1.09 (lH, m) and 0.~4 (6H, m) ppm.
FAB HRMS exact mass calcd for C39H47N6O4S 695.33795 (MH,
found 695.33893.
EXAMPLE 12
Preparation of N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -phenylmethyl-glycyl-methionine
bis trifluoroacetate
Step A: Preparation of N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-
5-ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -phenylmethyl-
glycvl-methionine methyl ester bis trifluoroacetate
To a solution of 1-(2-naphthylmethyl)-lH-imidazol-5-
ylacetic acid hydrochloride (prepared in Fx~mI)le 4, 75 mg, 0.25 mmol),
N-[2(S)-amino-3(S)-methylpentyl] -N-phenylmethyl-glycyl-methionine
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methyl ester bis hydrochloride (prepared analogously to 10, 112 mg,
0.248 mmol) and 3-hydroxy-1,2,3-benzotriazin-4(3H)-one (HOOBT, 44
mg, 0.27 mmol) in dimethylformamide (5 ml) was added 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 52 mg,
0.272 mmol) and triethyl~mine (171 ,ul, 1.23 mmol) and the suspension
stirred for 3 days. After this time, sat. aq. sodium bicarbonate (10 ml)
and water (10 ml) was added and the mixture was extracted with ethyl
acetate (2 x 50 ml). The combined extracts were washed with brine (20
ml) and the solvent evaporated in vacuo. Purification by preparative
HPLC (chromatography method A ) gave, after lyophili7~tion, the title
compound.
lH NMR (CD30D, 400 MHz) ~ 8.94 (lH, s), 7.93 (lH, d, J=8.5 Hz),
7.88 (2H, m), 7.81 (lH, s), 7.55 (5H, m), 7.43 (4H, m), 5.68 (2H, s), 4.60
(lH, m), 4.46 (lH, dd, J=4.5 Hz), 4.27 (lH, d, J=13 Hz), 4.14 (lH, m),
3.95 (lH, d, J=15.5 Hz), 3.85 (lH, d, J=15.5 Hz), 3.83 (2H, s), 3.67 (3H,
s),3.48(1H,d,J=13Hz),3.24(1H,d,J=13Hz),2.40(1H,m),2.31 (lH,
m), 2.00 (lH, m), 1.96 (3H, s), 1.85 (lH, m), 1.57 (lH, m), 1.44 (lH, m),
1.19 (lH, m), 0.93 (3H, d, J=6.7 Hz) amd 0.91 (3H, t, J=7 Hz) ppm.
Anal. Calcd for C37H47N504S-2.~5 TFA-0.40 H2O: C, 51.80; H, 5.16;
N, 7.07. Found: ~, 51.80; H, 5.14; N, 7.31.
Step B: Preparation of N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-
5-ylacetyl)amino-3 (S)-methylpentyl] -N- 1 -phenylmethyl-
glycyl-methionine bis trifluoroacetate
Following the procedure described in Example 9, Steps B,
but sub~liluli~g the methyl ester from Step A provided the title
compound.
lH NMR (CD30D, 400 MHz) ~ 8.92 (lH, s), 7.93 (lH, d, J=8.6 Hz),
7.87 (2H, m), 7.78 (lH, s), 7.55 (3H, m), 7.43 (2H, m), 7.39 (lH, d,
J=8.4Hz), 7.35 (3H, m), 5.67 (2H, s), 4.46 (lH, dd, J=4.5 Hz), 4.41-3.90
(lH, m), 4.11 (lH, m), 4.00 (lH, m), 3.75 (2H, m), 3.64 (2H, m), 3.20
(lH, m), 2.98 (lH~ m), 2.43 (lH, m), 2.35 (lH, m), 2.08 (lH, m), 1.97
(3H, s), 1.91 (lH, m), 1.54 (lH, m), 1.40 (lH, m), 1.15 (lH, m) and 0.89
(6H, m) ppm.
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Anal. Calcd for C36H45N504S-2.70 TFA-0.70 H20: C, 51.57; H, 5.13;
N, 7.26. Found: C, 51.54; H, 5.11; N, 7.43.
FAB HRMS exact mass calcd for C36H46N504S 644.327052 (MH+),
found 644.326203.
s
EXAMPLE 13
Preparation of N-[2(S)-(1 -(2-Naphthylmethyl)- 1 H-imidazol-5-
ylethyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-
10 methionine bis trifluoroacetate
StepA: Preparationof N-Methoxy-N-methyl-1-(2-naphthylmethyl)-
1 H-imidazol-5-ylacetamide
To a solution of 1-(2-naphthylmethyl)-lH-imidazol-5-
15 ylacetic acid hydrochloride (prepared in Example 4, 0.819 mg, 2.70mmol) in dimethylformamide (15 ml ) was added sequentially N, O-
dimethylhydroxyl~mine hydrochloride (293 mg, 3.0 mmol), 3-hydroxy-
1,2,3-benzotriazin-4(3H)-one (HOOBT, 489 mg, 3.0 mmol), 1-(3-
dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC, 575
20 mg, 3.0 mmol) and triethylamine (1.67 ml, 12.0 mmol) and the resulting
Lule stirred at room temperature for 18 h. Saturated aq. sodium
bicarbonate (30 ml) and water (30 ml) were added and the mixture was
extracted with methylene chloride (2 x 50 ml). The combined organic
extracts were washed with brine (50 ml) and the solvent evaporated in
2s vacuo. The residue was purified by flash chromatography (2-4%
methanol/methylene chloride gradient elution) to provide the title
compound as an oil.
1H NMR (CDCl3, 400 MHz) ~ 7.80 (2H, m), 7.74 (lH, m), 7.56 (lH, s),
7.47 (3H, m), 7.22 (lH, d, J=8.6 Hz), 6.97 (lH, s), 5.37 (2H, s), 3.58 (2H,
30 s),3.51 (3H,s)and3.12(3H,s)ppm.
Step B: 1 -(2-Naphthylmethyl)- 1 H-imidazol-5-vlacetaldehyde (30)
To a suspension of lithium al~ llll hydride (40.8 mg, 1.07
mmol) in tetrahydrofuran (5 ml) at -45C was added a solution of N-
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methoxy-N-methyl- 1 -(2-naphthylmethyl)- 1 H-imidazol-5-ylacetamide
(243 mg, 0.895 mmol) in tetrahydrofuran (5 ml) via c~nn~ at such a rate
to m~int~in the temperature at ~-35C. After the addition was complete,
the reaction was allowed to warm to +5C and then recooled to -35C.
5 To this solution was added a solution of potassium bisulfate (272 mg) in
water (1 ml). The mixture was stirred for 30 min at room temperature
and then filtered through celite. The celite pad was washed with ethyl
acetate (25 ml). The combined filtrates were washed with sat. sodium
bicarbonate (10 ml) and then water (10 ml). The organic layer was
o dried(MgSO4), filtered and evaporated in vacuo to give 30 as a clear oil.
This material was used as is in the next step.
lH NMR (CDC13, 400 MHz) o 9.50 (lH, t, J=2 Hz), 7.85-7.70 (3H, m),
7.64 (lH, s), 7.53-7.40 (3H, m), 7.16 (lH, d, J=12 Hz), 7.06 (llI, s), 5.20
(2H, s) and 3.53 (2H, m) ppm.
Step C: Preparation of N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-
5-ylethyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
glycyl-methionine methyl ester bis trifluoroacetate
To a solution of 1-(2-naphthylmethyl)-lH-imidazol-5-
20 ylacetaldehyde (116.~s mg, 0.465 mmol) and N-[2(S)-amino-3(S)-
methylpentyl]-N-naphthylmethyl-glycyl-methionine methyl ester bis
hydrochloride (lO, 297 mg, 0.558 mmol) in 1,2-dichloroethane (10 ml)
and dimethylform~mide (5 ml) was added 3A molecular sieves (500 mg)
and sodium triacetoxyborohydride (473 mg, 2.23 mmol). This mixture
25 was stirred at room temperature for 18 h. After this time, the mixture was
filtered through a sintered glass funnel. The filtrate was diluted with
methylene chloride (100 ml) and washed with sat. sodium bicaIbonate
(50 ml). The organic layer was dried over magnesium sulfate, filtered
and the solvent was evaporated in vacuo. The residue was purified first
30 by flash chromatography eluting with 2-5% methanol/methylene chloride
and then by preparative HPLC (chromatography method A) to provide
the title compound as a white foam.
1H NMR (CD30D, 400 MHz) ~ 9.05 (lH, s), 8.10 (lH, d, J=7.5 Hz),
8.02-7.79 (SH, m), 7.75 (lH, s), 7.65-7.27 (7H, m), 7.21 (lH, s), 5.59
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(2H,s),4.65(1H,dd,J=4.7and9.4Hz),4.31 (lH,d,J=13Hz),4.17(1H,
d, J=13 Hz), 3.69 (3H, s), 3.65 (lH, d, J=17 Hz), 3.55 (lH, d, J=17 Hz),
3.00 (lH, dd, J=3.5 and 14 Hz), 2.93-2.42 (6H, m), 2.33 (lH, m), 2.23
(lH, m), 2.13 (lH, m), 2.06 (3H, s), 1.96 (lH, m), 1.41 (lH, m), 1.07
(2H, m), 0.75 (3H, d, J=6.5 Hz) and 0.70 (3H, t, J=7.5 Hz) ppm.
FAB HRMS exact mass calcd for C41H52N5O3S 694.37909 (MH+),
found 694.37959.
Step D: Preparation of N-[2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-
5-ylethyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-
~lvcyl-methionine bis trifluoroacetate
Following the procedure described in Fx~mple 2, Steps D,
but Sub~liLulillg the methyl ester from Step C provided the title
compound.
H NMR (CD3OD, 400 MHz) ~ 8.95 (lH, s), 8.09 (lH, d, J=7.7 Hz),
7.94 (lH, d, J=8.5 Hz), 7.93-7.78 (4H, m), 7.73 (lH, s), 7.62-7.24 (7H,
m),7.17(1H,s),5.56(2H,s),4.61 (lH,dd,J=4.3and lOHz),4.31 (lH,
d, J=13 Hz), 4.14 (lH, d, J=13 Hz), 3.65 (lH, d, J=17 Hz), 3.55 (lH, d,
J=17 Hz), 2.99 (lH, d, J=15 Hz), 2.91-2.43 (6H, m), 2.25-1.91 (4H, m),
2.06 (3H, s), 1.33 (lH, m), 1.01 (2H, m), 0.72 (3H, d, J=6.7 Hz) and 0.65
(3H, t, J=7.5 Hz) ppm.
FAB HRMS exact mass calcd for C40H50N503S 680.36344 (MH+),
found 680.36282
EXAMPLE 14
Preparation of 2(S)-[N-2(S)-(1-(2-Naphthylmethyl)-lH-imidazol-5-
ylacetyl)amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-methionine
sulfone methyl ester hydrochloride
Step A: Preparation of N-(oc-chloroacetyl)-L-isoleucinol
To a stirred solution of L-isoleucinol (20 g, 0.17 mol) and
triethyl~mine (28.56 ml, 0.204 mol) in CH2Cl2 (500 ml) at -78C was
added chloroacetyl chloride (16.3 ml, 0.204 mol) over 5 millules. The
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cooling bath was removed and the solution allowed to warm to -20C.
The mixture was diluted with EtOAc and washed sequentially with 1 M
HCl, and brine and dried (Na2S04). Evaporation in vacuo afforded the
title compound
Rf = 0.3 CH2C12: MeOH (95:5);
lHNMR(CDC13)~ 6.80(1H,brd,J=SHz),4.10(2H,s),3.84(1H,m),
3.79 (2H, m), 2.65 (lH, brs), 1.72 (lH, m), 1.55 ~lH, m), 1.17 (lH, m),
0.96 (3H, d, J = 6Hz) 0.90 (3H,t, J=6 Hz).
Step B: Preparation of 5(S)-[l(S)-methyl]propyl-2,3,5,6-tetra-hydro-
4H- 1.4-oxazin-3-one .
To a stirred solution of N-(o~-chloroacetyl)-L-isoleucinol
(68, 7.4 g, 0.038 mol) in THF (125 ml) under argon at 0C was slowly
added sodium hydride (2.2 g of a 60% dispersion in mineral oil, 0.055
mol) with concomitant gas evolution. After completing the addition, the
mixture was warmed to room temperature (R.T.) and stirred for 16 hr.
Water (2.8 ml) was added and the solvents evaporated in vacuo. The
residue was dissolved in CHC13 (70 ml) and washed with saturated NaCl
solution. The organic layer was dried (Na2S04) and evaporated in
vacuo. The residue was chromatographed using silica gel eluting with
CH2Cl2:MeOH (96:4) to afford the title compound as a white solid.
Rf = 0.35 CH2Cl2:MeOH (95:5);
lHNMR (CDCl3)~6.72(1H,brs),4.20(1H,d,J= 14.5Hz),4.10(1H,
d,J=14.5Hz),3.88(1H,dd,J=9and3.5Hz),3.58(1H,dd,J=9and
6.5 Hz), 3.45 (lH, brqt, J = 3.5 Hz), 1.70-1.45 (2H, m), 1.34 - 1.15 (lH,
m),0.96(3H,t,J=6.5Hz),0.94(3H,d,J=6.5Hz).
Step C: Preparation of N-(tert-butoxycarbonyl)-5(S)-[l (S)-
methyllpropyl-2.3.5.6-tetrahydro-4H- 1,4-oxazin-3-one.
5(S)-[1 (S)-Methyl]propyl-2,3,5,6-tetrahydro 4H- 1,4-oxazin-
3-one (12.2 g, 0.0776 mol) and DMAP (18.9 g, 0.155 mol) were
dissolved in methylene chloride (120 ml) under argon at room
temperature. Boc anhydride (33.9 g, 0.155 mol) was added to the stirred
solution in one portion, with concomitant gas evolution and the mixture
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was stirred at for 16 hr. The solvent was evaporated in vacuo and the
residue was taken up in ethyl acetate and washed sequentially with 10%
citric acid, 50% NaHCO3 and finally brine. The organic extract was
dried (Na2SO4) and evaporated in vacuo. Chromatography of the
residue over silica gel eluting with 20% EtOAc in hexanes afforded the
title compound as a white solid.
Rf = 0.75 EtOAc:hexanes (20:80); mp 59-60C
Anal. Calcd for C13H23O4N: C, 60.68, H,9.01; N, 5.44. Found: C,
60.75; H, 9.01; N, 5.58.
H NMR (CDCl3) ~ 4.25 (lH, d, J = 15 Hz), 4.15 (lH, d, J = 15 Hz),
4.15 - 4.00 (2H, m), 3.73 (lH, dd, J = 10 and 2 Hz), 1.88 (lH, qt, J = 6
Hz), 1.55 (9H, s), 1.50 - 1.36 (lH, m), 1.35 - 1.19 (lH, m), 1.00 (3H, d, J
= 6 Hz), 0.95 (3H, d, J = 6.5 Hz).
Step D: Preparation of N-(tert-Butoxycarbonyl)-2(S)-benzyl-5(S)-
[1 (S)-methyl]propyl-2,3,5,6-tetrahydro-4H- 1,4-oxazin-3-
one
A solution of N-(tert-butoxycarbonyl)-5(S)-[l(S)-
methyl]propyl-2,3,5,6-tetrahydro-4H-1,4-oxazin-3-one (5.75 g, 22.3
mmol) in DME (100 ml) under argon was cooled to -60C. The cold
solution was transferred via canula to a second flask cont~ining sodium
bis(trimethylsilyl)amide (24.58 ml of a lM solution in THF, 24.58 mmol)
at -78C under argon. After stirring for 10 minlltes, benzyl bromide (2.25
ml, 19.0 mmol) was added over 5 minlltes and the resulting mixture was
stirred at -78C for 3 hours. After this time, the reaction mixture was
transferred via c~nn7ll~ to another flask cont~ining sodium
bis(trimethylsilyl)amide (24.58 ml of a lM solution in THF, 24.58 mmol)
at -78C, under argon. After stirring for a further 5 lll~llUL~S, the reaction
was quenched by the addition of saturated aqueous ammonium chloride
solution (24.6 ml) and allowed to warm to room temperature. This
mixture was diluted with brine (50 ml) and water (20 ml) and then
extracted with ethyl acetate (2 x 100 ml). The organic extracts were
washed with brine (50 ml) and evaporated in vacuo to afford an oil.
Chromatography of the residue over silica gel (230-400 mesh, 300 g)
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eluting with 10-20% ethyl acetate in hexanes afforded the title compound
as a clear oil.
Rf = 0.25 EtOAc:Hexanes (20:80);
lH NMR (CDC13) ~ 7.35 - 7.15 (5H, m), 4.31 (lH, dd, J = 6 and 2 Hz),
4.03(1H,d,J= 12Hz),3.88(1H,dd,J=6andlHz),3.66(1H,dd,J=
12 and 2 Hz), 3.29 (lH, dd, J = 12 and 3 Hz), 1.54 (9H, s), 3.12 (lH, dd,
J = 12 and 7 Hz), 1.47 (lH, m), 1.25 (lH, m), 1.10 (lH, m), 0.83 (3H, d, J
=6Hz),0.80(3H,t,J=6Hz).
Step E: Preparation of N-(tert-butoxycarbonyl)-2(S)-[2(S)~amino-
3tS)-methyllpentyloxy-3-phenyl-propionic acid
To a stirred solution of N-(tert-butoxycarbonyl)-2(S)-benzyl-
5(S)-[1 (S)-methyl] -propyl-2,3,5,6-tetrahydro-4H- 1,4-oxazin-3-one (5.1 g,
14.7 mrnol) in THF (150 ml) and water (50 ml) at 0C was added
hydrogen peroxide (15 ml of a 30% aqueous solution, 132 mmol) and
lithium hydroxide (3.0 g, 63.9 mmol). After stirring for 30 minutes, the
reaction was quenched with a solution of sodium sulfite (28.25 g, 0.224
mol) in water (70 ml). The THF was evaporated in vacuo and the
aqueous phase was acidified to pH 3-4 by addition of 10% citric acid
solution and extracted with EtOAc. The organic extracts were dried
(Na2S04), evaporated in vacuo and the residue purified by
chromatography over silica gel eluting with 4% MeOH in CH2Cl2 to
give 2(S)-benzyl-5(S)-[1 (S)-methyl]propyl-2,3,5,6-tetrahydro-4H- 1,4-
oxazin-3-one and then with 20% MeOH in CH2Cl2 to afford the title
compound as a white solid (pet ether, mp 68-70C).
Rf = 0.4 MeOH:CH2Cl2 (5:95) + 0.3% AcOH;
lH NMR (d6 DMSO) ~ 7.35 - 7.10 (5H, m), 6.68 (lH, br, s), 3.75 (lH,
dd, J = 7.5 and 2.5 Hz) 3.54 (lH, m), 3.5 - 3.2 (2H, m) 2.99 (lH, dd, J =
12.5 and 2.5 Hz), 2.75 (lH, dd, J = 12.5 and 7.5 Hz), 1.50 - 1.35 (1 lH,
m),0.98(1H,sept,J=6Hz),0.78(3H,t,J=6Hz),0.65(3H,dl,J=6
Hz);
FAB MS 366 (MH+) 266 (MH2+ - CO2tBu).
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Step F: Preparation of N-(tert-Butoxycarbonyl)-2(S)-[2(S)-amino-
3(S)-methyl]-pentyloxy-3-phenyl-propionyl-methione
sulfone methvl ester
The title compound was prepared by EDC coupling of N-
(tert-butoxycarbonyl)-2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-
phenylpropionic acid with methionine sulfone methyl ester.
1H NMR (CD30D) ~ 0.80 (3H, d, J=6 Hz), 0.88 (3H, t, J=6 Hz), 1.12
(lH, m), 1.40-1.55 (lH,m), 1.47 (9H, s), 2.10 (lH, m), 2.32 (lH, m),
2.80-3.10 (4H,m), 2.93 (3H, s), 3.40 (lH,m), 3.5-3.7 (2H, m), 3.74 (3H,
s), 4.01 (H, m), 4.60 (H, m), 6.60 (H, d, J=8 Hz), 7.25 (5H, m).
Step G: Preparation of 2(S)-[2(S)-Amino-3(S)-methyl]pentyloxy-3-
phenylpropionyl-methionine sulfone methyl ester
hydrochloride
N-(tert-butoxycarbonyl-2(S)-[2(S)-amino-3(S)-
methyl]pentyloxy-3-phenylpropionyl-methionine sulfone methyl ester
was treated with HCI gas in ethyl acetate and the solvent was evaporated
in vacuo to afford the title compound.
lH NMR (CD30D) ~ 0.85 (3H, d, J=6 Hz), 0.94 (3H, t, J=6 Hz), 1.20
(lH,m), 1.52(1H,m), 1.72(1H,m),2.14(1H,m),2.38(1H,m),2.98
(3H, s), 2.90-3.20 (4H, m), 3.25 (lH, m), 3.57 (lH, dd, J=12 and 6 Hz),
3.73 (lH, dd, J=12 and 9 Hz), 3.78 (3H, s), 4.15 (lH, m), 4.63 (lH, d,
J=8.5 Hz), 7.30 (5H, m).
Step H: Preparation of 2(S)-[N-2(S)-(1-(2-Naphthylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methyl]pentyloxy-3-
phenylpropionyl-methionine sulfone methyl ester
hydrochloride
To a solution of 1-(2-Naphthylmethyl)-lH-imidazol-5-
ylacetic acid hydrochloride (prepared in Example 4, 67 mg, 0.21 mmol),
2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-methionine
sulfone methyl ester hydrochloride (100 mg, 0.209 mmol) and 3-
hydroxy- l ,2,3-benzotriazin-4(3H)-one (HOOBT, 37.5 mg, 0.209 mmol)
in dimethylform~mide (4 ml) was added 1-(3-dimethylaminopropyl)-3-
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ethylcarbodiimide hydrochloride (EDC, 44 mg, 0.21 mmol) and
triethyl~mine (109 ul, 0.78 mmol) and the suspension stirred overnight.
After this time, sat. aq. sodium bicarbonate (7 ml) was added and the
resulting precipitate filtered. The precipitate was partitioned between
water (25 ml) and methylene chloride (50 ml). The organic extract was
evaporated in vacuo. The residue was purified by flash chromal:ography
eluting with 2-3% methanol/methylene chloride gradient to provide a
gum. The gum was dissolved in methanol (5 ml) and treated with
gaseous hydrogen chloride to pH=2 and the solution was evaporated in
vacuo. The resulting gum was dissolved in methanol (2 ml) and water
(20 ml) and lyophilized to give the title compoundas a white foam.
lH NMR (CD30D, 400 MHz) ~ 8.93 (lH, s), 8.35 (lH, d, J=8.7 Hz),
8.14 (lH, d, J=8.7 Hz), 7.94 (lH, d, J=8.6 Hz), 7.92-7.83 (2H, m), 7.77
(lH, s), 7.58-7.49 (3H, m), 7.38 (lH, d, J=8.4 Hz), 7.23-7.10 (SlH, m),
5.62 (lH, d, J=15.5 Hz), 5.61 (lH, d, J=15.5 Hz), 4.56 (lH, m), 4.05 (lH,
dd, J=4.0 and 7.4 Hz), 3.90 (lH, m), 3.70 (2H, s), 3.66 (3H, s), 3.57 (lH,
dd, J=3.5 and 9.9 Hz), 3.47 (lH, dd, J=7.0 and 9.9 Hz), 3.04 (lH, dd,
J=4.0 and 14.1 Hz), 2.96 (lH, m), 2.91 (lH, dd, J=7.5 and 14.1 lEIz), 2.90
(3H, s), 2.80 (lH, m), 2.27 (lH, m), 2.09 (lH, m), 1.50 (lH, m), 1.43
(lH, m), 1.07 (lH, m), 0.84 (3H, t, J=7.4 Hz) and 0.77 (3H, d, J=6.7 Hz)
ppm.
Anal. Calcd for C37H46N4O7S-2.3 HCl: C, 57.36; H,6.28; N, 7.23.
Found: C, 57.40, H, 6.20; N, 7.38.
FAB HRMS exact mass calcd for C37H47N407S 691.316547 (MH+),
found 691.316460.
EXAMPLE 15
Preparation of 2(S)-[N-2(S)-(1-(2-Naphthylmethyl)-lH-imi~1~7ol-5-
ylacetyl)amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-methionine
sulfone trifluoroaçetate
Following the procedure described in Example 9, Step B,
but substituting the methyl ester from Example 14 provided the title
compound.
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lH NMR (CD30D, 400 MHz) ~ 8.93 (lH, s), 8.27 (lH, d, J=8.3 Hz),
8.10 (lH, d, J=9.3 Hz), 7.94 (lH, d, J=8.6 Hz), 7.92-7.83 (2H, m), 7.75
(lH, s), 7.57-7.52 (2H, m), 7.50 (lH, s), 7.37 (lH, d, J=8.6 Hz), 7.23-
7.11 (SH, m), 5.60 (lH, d, J=15 Hz), 6.59 (lH, d, J=15 Hz), 4.54 (lH, m),
4.03 (lH, dd, J=4.1 and 7.9 Hz), 3.91 (lH, m), 3.69 (lH, d, J=16.7 Hz),
3.66 (lH, d, J=16.7 Hz), 3.56 (lH, dd, J-3.4 and 10.3 Hz), 3.45 (lH, dd,
J=7.0 and 9.7 Hz), 3.04 (lH, dd, J=4.2 and 15.1 Hz), 3.00 (lH, m), 2.94-
2.85(1H,m),2.89(3H,s),2.80(1H,m),2.30(1H,m),2.09(1H,m),
1.50 (lH, m), 1.43 (lH, m), 1.07 (lH, m), 0.83 (3H, t, J=6.4 Hz) and 0.75
o (3H, d, J=6.7 Hz) ppm.
Anal. Calcd for C36H44N4O7S-2.10 TFA-0.90 H2O: C, 51.78; H,5.18;
N,6.01. Found:C,51.78;H,5.17;N,6.42.
FAB HRMS exact mass calcd for C36H4~N4O7S 677.300897 (MH+),
found 677.299827.
EXAMPLE 16
Preparation of 2(S)- [N-2(S)-(1 -(2-Naphthylmethyl)- 1 H-imidazol-5-
ylethyl)amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-methionine
methyl ester bis trifluoroacetate
Step A: Preparation of 2(S)-[2(S)-t-butoxycarbonylamino-3(S)-
methyl]-pentyloxy-3-phenylpropionyl-methionine methyl
ester
The title compound was prepared in the same fashion as that
described in Example 14, Step F, using methionine methyl ester in place
of methionine sulfone methyl ester.
NMR (CD30D) ~ 0.78 (3H, d, J=6 Hz), 0.89 (3H, t, J=6 Hz). 1.11 (lH,
m), 1.40-1.60 (2H, m), 1.47 (9H, s), 1.90-2.10 (2H,m), 2.06 (3H, s),
2.20-2.40 (2H, m), 2.90 (lH, dd, J=14.7 and 5.0 Hz), 3.05 (H,dd, J=14.5
and 3.0 Hz), 3.38 (lH, dd, J=8.6 and 7.0 Hz), 3.50-3.60 (2H, m), 3.71
(3H, s), 3.97 (lH, dd, J=7.5 and 4.0 Hz), 4.60 (lH, m), 6.60 (lH, d, J=10
Hz), 7.24 (SH, m).
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Step B: Preparation of 2(S)-[2(S)-amino-3(S)-methyl]-pentyloxy-3-
phenvlpropionyl-methionine methyl ester hydrochloride
The product of Step A was converted to the title compound
using the method of Example 14, Step G.
lH NMR (CD30D) ~ 0.84 (3H, d, J=6 Hz), 0.93 (3H, t, J=6 Hz), 1.20
(lH, m), 1.45-1.60 (lH, m), 1.70 (lH, m), 1.80-2.20 (2H, m) 2.08 (3H, s),
2.50-2.30 (2H, m), 2.98 (lH, dd, J=14.7 and 5 Hz), 3.11 (lH, dd, J=14.5
and 3.0 Hz), 3.20-3.30 (lH, m), 3.57 (lH, m), 3.70 (lH, m), 3.73 (3H, s),
4.12 (H, dd, J=8.6 and 6.0 Hz), 4.60 (lH, m), 7.30 (5H, m).
Step C: Preparation of 2(S)-~N-2(S)-(1-(2-Naphthylmethyl)-lH-
imidazol-5-ylethyl)amino-3(S)-methyl]pentyloxy-3-
phenylpropionyl-methionine methyl ester bis
trifluoroacetate
Following the procedure described in Example 13, Step C,
but sub~liLuLillg 1-(2-naphthylmethyl)-lH-imidazol-5-ylacetaldehyde (30)
and 2(S)-[2(S)-amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-
me~ionine methyl ester hydrochloride, the title compound was obtained.
lH NMR (CD3OD, 400 MHz) ~ 8.95 (lH, s), 7.96 (lH, d, J=8.5 Hz),
7-89 (2H, m), 7.79 (lH, s), 7.55 (2H, m), 7.47 (lH, s), 7.38 (lH, d, 8.4
Hz), 7.21 (4H, m), 7.15 (lH, m), 5.65 (2H, s, 4.63 (lH, dd, J=4.4 and
19.5 Hz), 4.15 (lH, dd, J=4.3 and 18.7 Hz), 3.67 (3H~ s), 3.57 (2H, m),
3.43-3.15 (2H, m), 3.11-3.00 (4H, m), 2.88 (lH, dd, J=9 and 14-.4 Hz),
2.51 (lH,m),2.40(1H,m),2.10(1H,m),2.03(3H,s), 1.95(11:I,m),
1.68 (lH, m), 1.35 (lH, m), 1.09 (lH, m), 0.86 (3H, t, J=7.2 Hz) and 0.74
(3H, d, J=6.9 Hz) ppm.
Anal. Calcd for C37H4gN404S~2.45 TFA: C, 54.45; H,5.50; N, 6.06.
Found: C, 54.37; H, 5.51; N, 6.15.
FAB HRMS exact mass calcd for C37H49N404S 645.34745 (MH+),
found 645.34518.
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EXAMPLE 17
Preparation of 2(S)-[N-2(S)-(1 -(2-Naphthylmethyl)- l H-imidazol-5-
ylethyl)amino-3(S)-methyl]pentyloxy-3-phenylpropionyl-methionine bis
trifluoroacetate
Following the procedure described in Example 2, Step D,
but substituting the methyl ester from Example 16 provided the title
compound.
H NMR (CD30D, 400 MHz) ~ 8.89 (lH, s), 7.95 (lH, d, J=8.5 Hz),
7-93-7-84 (2H, m), 7.77 (lH, s), 7.58-7.51 (2H, m), 7.45 (lH, s), 7.37
(lH, dd, J=1.7 and 8.3 Hz), 7.26-7.17 (4H, m), 7.15 (lH, m), 5.65 (2H,
s), 4.59 (lH, dd, J=4.5 and 9.4 Hz), 4.14 (lH, dd, J=3.8 and 8.9 Hz), 3.56
(2H, d, J=3.8 Hz), 3.37-2.96 (6H, m), 2.88 (lH, dd, J=8.8 and 14.2 Hz),
2.52 (lH, m), 2.41 (lH, m), 2.16 (lH, m), 2.03 (3H, s), 1.97 (lH, m),
1.66 (lH, m), 1.32 (lH, m), 1.08 (lH, m), 0.85 (3H, t, J=7.1 Hz) and 0.74
(3H, d, J=7.1 Hz) ppm.
Anal. Calcd for C36H46N404S-2.95 TFA-1.00 H2O: C, 51.08; H,5.21;
N, 5.69. Found: C, 51.07; H, 5.22; N, 5.83.
FAB MS calcd for C36H47N4O4S, 631 (MH+), found 631.
EXAMPLE 1~
Preparation of N-r2(S)-(l-methyl-imidazol-4-yl acetyl)amino-3(S)-
methylpentyl~-N-(l-naphthylmethyl)-glycyl-methionine methyl ester
trifluoroacetate salt
l-Methyl-4-imidazole acetic acid (0.070 g, 0.395 mmol),
dissolved in DMF (5 mL), was treated with HOBT (0.053 g, 0.040
mmol), EDC (0.075 g, 0.395 mmol), and N-[2(S)-amino-3-
methylpentyl)-N-(l-naphthylmethyl)-glycyl-methionine methyl ester
hydrochloride (10, 0.175 g, 0.395 mmol). The pH was adjusted to 7.5
with Et3N (0.055 mL, 0.395 mmol) and the mixture was stirred at
ambient temperature for 72 h. The mixture was concentrated and the
residue was partitioned between EtOAc (30 mL) and saturated NaHCO3
solution (25 mL). The aqueous layer was extracted with EtOAc (2x20
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mT .). The combined organic layer was washed with brine (lx25 mL),
dried (Na2SO4), and evaporated in vacuo to give a crude prodl1ct which
was purified by chromatography (silica gel, eluting with 99:1 to 97:3
CH2Cl2:MeOH) to give the amine. This material was converted to the
5 trifluroracetate salt by dissolving in 0.1% TFA in H20 and Iyophili7~tion
to give the title compound. lH NMR (CD30D) ~ 8.72 (lH, s), 8.30-
8.20 (lH, m), 8.00-7.90 (2H, m), 7.45-7.70 (4H, m), 7.34 (lH, s), 4.80-
4.65(1H,m),4.60-4.40(2H,m),4.20-4.10(1H,m),3.86(3H,s),3.70
(3H, s ), 3.85-3.50 (4H, m ), 3.40-3.30 (lH, m), 3.20-3.05 (lH, m), 2.40-
2.20 (2H, m), 2.00 (3H, s ), 2.00-1.90 (lH, m), 1.82-1.65(1H, m), 1.65-
1.52 (lH, m), 1.50-1.35 (lH, m), 1.25-1.07 (lH, m), 1.00-0.85 (6H, m).
Anal. Calcd for C31H43N504S-3 TFA: C, 48.10; H, 5.02; N, 7.58.
Found: C, 48.36; H, 5.30; N, 7.77.
EXAMPLE 19
Preparation of N-[2(S)-(1-methyl-lH-imidazoleacetyl) arnino -3(S)-
methylpentyll-N-( l -naphthylmethyl)-glycyl-methionine
N-[2(S)-(1-Methyl-4-imidazoleacetyl) amino-3(S)-
20 methylpentyl]-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
(prepared in Example 18, 0.081 g, 0.139 mmol) was dissolved in MeOH
(5 ml), cooled to 0C, and lN NaOH (0.557 ml, 0.557 mmol) was added.
The mixture was stirred at ambient temperature for 4 h and evaporated in
vacuo. The resulting residue was dissolved in H2O (5 ml) and
25 neutralized with lN HCl (0.557 ml, 0.557 mmol). The aqueous layer was
washed with EtOAc (3x10 ml). The organic layers were combined, dried
( Na2SO4), and evaporated in vacuo to give a crude product. Purification
by preparative HPLC (Vydac column eluting with acetonitrile/0.1 % TFA
in H2O gradient) and lyophili7~tion gave the title compound. 1H NMR
30 (CD30D) ~ 8.72 (lH, s), 8.31-8.23 (lH, m), 8.02-7.90 (2H, m), 7.70-
7.45 (4H, m), 7.35 (lH, s), 4.93-4.74 (lH, m), 4.58 (lH, d, J=13 Hz),
4.45-4.36 (lH, m), 4.20-4.10 (lH, m), 3.89 (3H, s~, 3.86-3.52 (4H, m),
3.45-3.30 (lH, m), 3.22-3.09 (lH, m), 2.45-2.20 (2H, m), 2.00 (3H, s),
=
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2.10-1.92 (lH, m), 1.83-1.68 (lH, m), 1.68-1.52 (lH, m), 1.52-1.37 (lH,
m), 1.26-1.08 (lH, m), 1.00-0.85 (6H, m).
Anal. Calcd for C30H41N5O4S-2.75 CF3CO2H: C, 48.38, H, 5.00; N,
7.95.
Found: C, 48.53; H, 5.05, N, 8.11.
EXAMPLE 20
Preparation of N-[2(S)-1-(2-naphthylmethyl)-lH-imidazol-5-
ylacetyl] amino-3 (S)-methylpentyl] -N-(cyclopropylmethyl)-
~lycylmethionine methyl ester bis trifluoroacetate salt
Step A: Preparation of N-[2(S)-t-Butoxycarbonylamino)-3-
methylpentyll-N-(cvclopropylmethyl)~lycine methyl ester
N-[2(S)-t-Butoxycarbonylamino)-3(S)-methylpentyl]glycine
methyl ester (6, 287.8 mg, 0.9980 mmol) was dissolved in 1,2-dichloro-
ethane (7.0 ml). 4A Molecular sieves (207 mg), cyclopropane-
carboxaldehyde (75 ml, 1.0 mmol), and sodium triacetoxyborohydride
(1.075 g, 5.072 mmol) were added. The mixture was stirred under argon
20 at ambient temperature for 16 h and filtered. The filtrate was diluted with
EtOAc (50 mL) and washed with saturated aq NaHCO3 (2 x 25 ml) and
saturated aq NaCl (25 mL). The organic layer was dried (Na2S04) and
evaporated in vacuo. The crude product was purified by chromatography
(silica gel, 1:19 to 1:9 EtOAc/CH2Cl2) to give the title compound. 1H
25 NMR (CDCl3, 400 MHz): ~ 4.85 (lH, br s), 3.69 (3H, s), 3.64-3.54 (lH,
m),3.70(1H,d,J= 18Hz),3.30(1H,d,J= 18Hz),2.74(1H,dd,J= 14
and 5 Hz), 2.57-2.42 (3H, m), 1.80-1.68 (lH, m), 1.50-1.36 (lH, m), 1.44
(9H, s), 1.15-1.02 (lH, m), 0.91 (3H, t, J=7 Hz), 0.86 (3H, d, J=7 Hz),
0.86-0.76 (lH, m), 0.54-0.43 (2H, m), 0.09 (2H, d, J=5 Hz).
Step B: Preparation of N-[2(S)-t-Butoxycarbonylamino)-3-
methvlpentyll -N-(cyclopropylmethyl)~lvcine
N-r2(S)-t-Butoxycarbonylamino)-3-methylpentyl] -N-
(cyclopropylmethyl)glycine methyl ester (268 mg, 0.783 mmol) was
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dissolved in MeOH (40 ml). After cooling to 0C under argon, lN aq
LiOH (1.0 ml, 1.0 mmol) was added. After stirring at ambient
temperature for 18 h, additional lN aq LiOH (1.0 ml, 1.0 mmol) was
added. After stirring at ambient temperature for 6 h, additional lN aq
LiOH (1.0 ml, 1.0 mmol~ was added. After stirring for 18 h at ambient
temperature, 1 N aq HCl (4.0 mL, 4 mmol) was added and the reaction
was evaporated in vacuo. The resulting residue was dissolved in H2O
(10 ml) and acidified with lN aq HCl to pH = 2. Residual methanol was
evaporated in vacuo and the rem~ining aqueous material lyophilized to
0 give the title compound. lH NMR (CD30D, 400 MHz): ~ 3.86-3.76
(2H, m), 3.62 (lH, d, J = 15 Hz), 3.47 (lH, br d), 3.28-3.14 (2H, m),
3.12-3.03 (lH, m), 1.64-1.43 (2H, m), 1.47 (9H, s), 1.26-1.10 (2H, m),
0.98-0.90 (6H, m), 0.80-0.68 (2H, m), 0.51-0.41 (2H, m).
Step C: Preparation of N-[2(S)-t-Butoxycarbonylamino)-3-
methylpentyl] -N-(cyclopropylmethyl)glycylmethionine
methyl ester
The title compound was prepared in the same fashion as that
described in Example 1, Step G, but using the compound described in
Step B.
1H NMR (CDC13, 400 MHz): ~ 8.02 (lH, br d), 4.78-4.68 (lH, m), 4.67
(lH, td, J = 9 and 6 Hz), 3.75 (3H, s), 3.70-3.60 (lH, m), 3.31 ~lH, d, J =
17Hz), 3.18 (lH, d, J = 17 Hz), 2.67 (lH, dd, J = 9 and 4 Hz), 2.54 (2H, t,
J = 8 Hz), 2.54-2.44 (2H, m), 2.43-2.35 (lH, m), 2.30-2.20 (lH, m), 2.16-
2-06 (lH, m), 2.10 (3H, s), 1.63-1.52 (lH, m), 1.50-1.40 (lH, m), 1.44
(9H, s), 1.17-1.05 (lH, m), 0.93 (3H, d, J = 8 Hz), 0.91 (3H, t, J = 8 Hz),
0.90-0.80 (lH, m), 0.56-0.46 (2H, m), 0.15 (2H, d, J = 6 Hz).
Step D: Preparation of N-[2(S)-Amino-3-methylpentyl)-N-
(cyclopropylmethyl)glycylmethionine methyl ester
hydrochloride
N-[2(S)-t-Butoxycarbonylamino)-3-methylpentyll-N-
(cyclopropylmethyl)-glycylmethionine methyl ester (22.8 mg, 0.0481
mmol) was dissolved in EtOAc (1.5 mL) and cooled to 0C. ~Cl was
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bubbled through the mixture until saturated. After 30 min, the mixture
- was evaporated in vacuo to give the title compound.
lH NMR (CD30D, 400 MHz): o 4.68 (lH, dd, J = 9 and 5 Hz), 4.2~
4.00 (2H, m), 3.74 (3H, s), 3.70-3.45 (2H, m), 3.40-3.00 (3H, m), 2.67-
2.51 (2H, m), 2.23-1.95 (2H, m), 2.10 (3H, br s), 1.87-1.86 (lH, m), 1.60-
1.49 (lH, m), 1.34-1.21 (lH, m), 1.20-1.10 (lH, m), 1.03 (3H, d, J = 7
Hz), 1.01 (3H, t, J = 7 Hz), 0.82-0.72 (2H, m), 0.50-0.40 (2H, m).
Step E: Preparation of N-[(2S)-1-(2-naphthylmethyl)-lH-imidazol-
5-ylacetyl]amino-(3S)-methylpentyl]-N-cyclopropylmethyl)-
~lvcylmethionine methyl ester bis trifluoroacetate salt
The title compound was prepared in the same fashion as that
described in Example 1, Step I, but using the compound prepared in
Step D.
lH NMR (CD30D, 400 MHz): ~ 8.93 (lH, s), 7.95 (lH, d, J = 9 Hz),
7.93-7.85 (2H, m), 7.80 (lH, s), 7.60-7.53 (3H, m), 7.42 (lH, dd, J = 9
and 2 Hz), 5.68 (2H, s), 4.69-4.45 (lH, m), 4.30-3.90 (3H, m), 3.90-3.80
(2H, m), 3.69 (3H, s), 3.60-3.45 (lH, m), 3.40-3.14 (3H, m), 2.60-2.40
(2H, m), 2.15-2.05 (lH, m), 2.03 (3H, s), 2.00-1.85 (lH, m), 1.60-1.52
(lH, m), 1-50-1.40 (lH, m), 1.25-1.15 (lH, m), 1.12-1.05 (lH, m), 0.98-
0.90 (6H, m), 0.80-0.68 (2H, m), 0.50-0.40 (2H, m).
FAB HRMS exact mass calcd for C34H4gNsO4S: 622.342702 (MH+);
found 622.343884.
EXAMPLE 21
Preparation of N-[(2~)- 1 -(2-naphthylmethyl)- 1 H-imidazol-5-
ylacetyl]amino-(3S)-methylpentyl] -N-(cyclopropylmethyl)-
~Iycvlmethionine bis trifluoroacetate salt
N-[(2S)-N-(2-Napthylmethyl) 1 H-imidazol-5-
ylacetyl]amino-(3S)-methylpentyl] -N-(cyclopropylmethyl)-
glycylmethionine methyl ester (19.8 mg, 0.0319 mmol) was dissolved in
MeOH (0.60 ml), cooled to 0C under argon, and treated with 1.0 N aq
LiOH (38 ml, 0.038 mrnol). After stirring at ambient temperature for 16
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h, the reaction was diluted with MeOH (1.5 ml) and purified by
preparative HPLC (chromatography method A) to give the title
compound as its bis trifluoroacetate salt after lyophili7~tion. lH NMR
(CD30D, 400 MHz): ~ 8.95 (lH, s), 7.95 (lH, d, J = 9 Hz), 7.94-7.85 A
(2H, m), 7.82 (lH, s), 7.62-7.52 (3H, m), 7.44 (lH, dd, J = 9 and 1 Hz),
5.60 (2H, s), 4.65-4.50 (lH, m), 4.23-4.05 (2H, m), 4.01-3.93 (lH, m),
3.89(1H,d,J= l9Hz),3.82(1H,d,J= l9Hz),3.52(1H,d,J= 14Hz),
3.30-3.05 (3H, m), 2.61-2.40 (2H, m), 2.20-2.10 (lH, m), 2.05 (3H, s),
2.00-1.89 (lH, m), 1.62-1.52 (lH, m), 1.50-1.40 (lH, m), 1.25 1.04 (2H,
m), 0.97 (3H, d, J = 7 Hz), 0.92 (3H, t, J = 7 Hz), 0.79-0.65 (2H, m),
0.50-0.40 (2H, m). Anal. Calcd for C33H45N504S-2.70 TFA~0.45
H2O: C, 49.93; H, 5.30; N, 7.58. Found: C, 49.90; H, 5.29; N, 7.92.
FAB HRMS exact mass calcd for C33H46N5O4S: 608.327052 (MH+);
found 608.326603.
EXAMPLE 22
Preparation of N-[2(S)-[(5(R,S)-Methylpyroglutamyl)amino]-3(S)-
methylpentyl]-N-(l-naphthylmethyl)-glycylmethionine methyl ester
trifluoroacetate salt-diastereomers A (31) and B (~2)
N-[2(S)-amino-3-methylpentyl)-N-(l -naphthylmethyl)-
glycyl-methionine methyl ester hydrochloride (10, 186.1 mg, 0.349
mmol) was dissolved in methylene chloride (3 mL). DL-2-Methyl-5-
pyrrolidone-2-carboxylic acid (K. Pfister III, W. J. T e~n7~, J. P. Conbere,
H. J. Becker, A. R. Mat_uk, and E. F. Rogers, J. Am. Chem. Soc., 77:697-
700 (1955), 50.2 mg, 0.351 mmol) was added followed by triethylamine
(270 mL, 1.94 mmol). The mixture was cooled to 0C under argon and
treated with bis(2-oxo-3-oxazolidinyl)phosphinic chloride (BOP-Cl,
133.3 mg. 0.5236 mmol). The reaction was stirred for 18 h at ambient
temperature, diluted with EtOAc (20 mL), washed with saturated aq
NaHCO3 (20 mL), saturated aq NaCl (20 mL), dried (Na2so4) and
evaporated in vacuo to give the crude product as a mi~Lur~ of
diastereomers. Purification by chromatography (silica gel, 1 :40
MeOH/CH2Cl2) gave the two diastereomeric products as an inseparable
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mixture. Separation of the diastereomers was accomplished through prep
plate chromatographies (silica gel, 3-5% MeOH/CH2Cl2) to give the
high Rf diastereomer (31) and the low Rf diastereomer (32) as colorless
residues. Final purification of each diastereomer was accomplished by
chromatography method A. Compounds 31 and 32 were obtained as the
trifluoroacetate salts by lyophili7~tion of appropriate column fractions.
31: lH NMR (CD30D, 400 MHz): ~ 8.25-8.17 (lH, m), 7.95-7.82 (2H,
m), 7.68-7.40 (4H, m), 5.10-2.80 (6H, m), 4.50-4.30 (lH, m), 4.10-3.95
(lH, m), 3.65 (3H, s), 2.60-0.90 (17H, m), 0.83 (3H, d, J = 7 Hz), 0.78
(3H,t,J=8Hz).
Anal. Calcd for C31H44N405S-l.10 TFA-0.10 H20: C, 56.01, H, 6.41;
N, 7.87. Found: C, 56.02; H, 6.29; N, 8.04.
FAB HRMS exact mass calcd for C31H45N4O5S: 585.311068 (MH+);
found 585.311153.
32: lH NMR (CD30D, 400 MHz): ~ 8.25-8.15 (lH, m), 7.95-7.81 (2H,
m), 7.65-7.38 (4H, m), 5.00-2.80 (6H, m), 4.42-4.28 (lH, m), 4.05-3.95
(lH, m), 3.63 (3H, s), 2.70-1.00 (17H, m), 0.85 (3H, br d, J = 7 Hz), 0.80
(3H, br t, J = 7 Hz). Anal. Calcd for C31 H44N405S- 1.05 TFA-0.20
H2O: C, 56.14; H, 6.47; N, 7.91. Found: C, 56.17; H, 6.47; N, 8.12.
FAB HRMS exact mass calcd for C3lH4sN4oss: 585.311068 (MH+);
found 585.311694.
EXAMPLE 23
Preparation of N-[2(S)-[(5(R,S)-methyl-pyroglutamyl)amino]-3(S)-
methylpentyl]-N-(l-naphthylmethyl)-glycylmethionine trifluoroacetate
salt.
N-[2(S)-[(5(R,S)-Methyl-pyroglutamyl)amino]-3(S)-
methylpentyl]-N-(l-naphthylmethyl)-glycyl-methionine methyl ester (31,
32.3 mg, 0.0552 mmol) was dissolved in MeOH (1.5 mL) under argon
and treated with 1.0 N aq LiOH (66 ~L, 0.066 mmol). The reaction was
stirred at ambient temperature for 18 h, treated with glacial acetic acid (2
drops), and purified by chromatography method A to give, after
lyophili7~tion, the title compound as a 2:1 mixture of diastereomers as
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their trifluoroacetate salts. lH NMR (CD30D, 400 MHz): ~ 8.29 (lH,
d, J = 8 Hz), 8.00-7.89 (2H, m), 7.78-7.45 (4H, m), 5.00-2.80 (8H, m),
2.60-1.00 (17H, m), 0.96-0.84 (6H, m).
Anal. Calcd for C30H42N4O5S-1.25 TFA-0.20 H2O: C, 54.45; H, 6.14;
N, 7.82. Found: C, 54.46; H, 6.14; N, 7.91.
FAB HRMS exact mass calcd for C30H43N4O5S: 571.295418 (MH+);
found 571.295373.
EXAMPLE 24
Preparation of N-[2(S)-[(5(R,S)-methylpyroglutamyl)amino]-3(S)-
methylpentyl] -N-( l -naphthylmethyl)-glycylrnethionine trifluoroacetate
salt
Following the procedure described in Example 23, but
substituting the methyl ester 32 from Example 22, the title compound was
prepared.
1H NMR (CD30D, 400 MHz): ~ 8.36-8.26 (lH, m), 7.97 (2H, br d, J =
8 Hz), 7.80-7.44 (4H, m), 5.00-3.00 (8H, m), 2.60-1.10 (17H, m), 0.99-
0.84 (6H, m).
20 Anal. Calcd for C3oH42N4oss-l.4o TFA-0.15 H20: C, 53.74; H, 6.01;
N, 7.64. Found: C, 53.73, H, 5.99; N, 7.74.
FAB HRMS exact mass calcd for C30H43N4O5S: 571.295418 (MH+);
found 571.296351.
EXAMPLE 25
Preparation of N-[2(S)-((N-methylpyroglutamyl)amino)-3(S)-
methylpentyl]-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
trifluoroacetate salt
N-methylpyroglllt~m~te [E. Hardegger and H. Ott, Helv.
Chim Acta, 38:312 (1955), 51 mg, 0.35 mmol)], dissolved in DMF (2.5
ml), was treated with HOBT (48 mg, 0.35 mmol), EDC (81 mg, 0.42
mmol), N-[2(S)-amino-3(S)-methylpentyl)-N-(l-naphthylmethyl)glycyl-
methionine methyl ester hydrochloride (10, 150 mg, 0.28 mmol), and
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triethyl~mine (0.079 ml, 0.56 mmol). The mixtl1re was stirred at room
temperature for 24 hours. The mixture was partitioned between ethyl
acetate and 10% citric acid solution and the organic phase was washed
three times with saturated NaHCO3, brine, and dried (MgSO4). The
5 solution was filtered through celite and evaporated in vacuo . The crude
product was chromatographed (5% MeOH in EtOAc) and further purified
by preparative HPLC (Waters PrepPak C-18 eluting with CH3CN/0.1%
TFA in H2O) to give, after lyophili7~tion, the title compound.
H NMR (CD30D) ~ 8.35(1H,d), 8.0(2H,m), 7.7(4H,m), 5.1(1H,m),
4-75(1H,m), 4.55(1H,m), 4.05(4H,m), 3.75(3H,s), 3.60(1H,m),
3.20(1H,m), 2.70(3H,s), 2.30(6H,m), 2.00(4H,m), 1.85(1H,m),
1.65(1H,m), 1.45(1H,m), 1.25(1H,m), 0.95(6H,m).
FAB MS calcd for C31H4sN4O5S 585 (MH+), found 585.
Anal. Calcd for C31H44N405S-1.35TFA-1.60H20: C, 52.73; H,6.38; N,
15 7.30.
Found: C, 52.75; H, 6.00; N, 7.70
EXAMPLE 26
20 Preparation of N-[2(S)-((N-methylpyroglutamyl)-amino)-3(S)-
methylpentyll-N-(1 -naphthylmethyl)-glycyl-methionine
N-[2(S)-((N-Methylpyroglutamyl)-amino)-3(S)-
methylpentyl]-N-( l -naphthylmethyl)-glycyl-methionine methyl ester
trifluoroacetate salt (prepared in E~cample 25, 112 mg, 0.19 mmol) was
25 dissolved in methanol (5 ml) and treated with 0.76 ml of lN LiOH. The
mixture was stirred for 4 hours at room temperature, then treated with
0.76 ml of lN HCl. The solvent was evacuated in vacuo. The crude
product was purified by preparative HPLC (Waters PrepPak C-18 eluting
with CH3CN/0.1% TFA in H20) to give, after lyophili7~tion, the title
30 compound.
lH NMR (CD30D) ~ 8.35 (lH,d), 8.00 (2H,m), 7.65 (4H,m), 5.10
(lH,m), 4.75 (lH,m), 4.50 (lH,m), 4.05 (4H,m), 3.60 (lH,m), 3.25
(lH,m), 2.70 (3H,s), 2.30 (6H,m), 2.05 (3H,s), 1.85 (2H,m), 1.60
(lH,m), 1.45 (lH,m), 1.20 (lH,m), 0.95 (6H,m).
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FAB MS calcd for C30H43N405S: 571 (MH+), found 571.
Anal. Calcd for C30H42N4O5S-1.60TFA-0.55H20: C, 52.25; H, 5.90;
N, 7.34. Found: C, 52.27; H, 5.92; N, 7.71.
EXAMPLE 27
Preparation of N-[2(S)-(N-formylprolylamino)-3(S)-methylpentyl]-N-(l-
naphthylmethyl)-glycyl-methionine methyl ester trifluoroacetate salt
N-formyl-L-proline [T. Sawayama, et al, Chem. Pharm.
10 Bull., 38 (2), 529-531 (1990), 44.3 mg, 0.31 mmol], dissolved in DMF (3
ml), was treated with HO~3T (46 mg, 0.34 mmol), EDC (81 mg, 0.42
mmol), N-~2(S)-amino-3-methylpentyl)-N-(l-naphthylmethyl)glycyl-
methionine methyl ester hydrochloride (10, 150 mg, 0.28 mmol), and
triethyl~mine (0.079 ml, 0.56 mmol). The mi~Lule was stirred at room
15 temperature for 72 h, then partitioned between ethyl acetate and 10%
citric acid solution. The organic extract was washed with saturated
NaHCO3 three times, then brine, and dried (MgSO4). After filtration
through celite and evaporation of solvent in vacuo., the crude product
was purified by preparative HPLC (Waters PrepPak C- 18 eluting with
20 CH3CN/0.1%TFA in H2O) to give, after lyophilization, the title
compound. lH NMR (CD30D) 8.35 (lH,m), 8.20 (lH,s), 8.00 (2H,m),
7.65 (4H,m), 5.10 (lH,m), 4.65 (2H,m), 4.10 (4H,m), 3.75 (3H,s), 3.60
(3H,m), 3.10 (lH,m), 2.40 (2H,m), 1.90 (8H,m), 1.55 (3H,m), 1.20
(lH,m), 0.90 (6H,m).
25 FAB MS calcd for C31H45N4O5S 585 (MH+), found 571.
Anal. Calcd for C31H44N405S-1.40TFA-0.20H20: C, 54.28; H, 6.11;
N,7.47.
Found: C, 54.25; H, 6.16, N, 7.69.
EXAMPLE 28
Preparation of N-[2(S)-(N-formylprolylamino)-3(S)-methylpentyl]-N-(l-
naphthylmethyl) -glycyl-methionine
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The procedure described in Fx~mple 26, sub~ illg the
methyl ester prepared in Example 27 was used to obtain the title
compound.
FAB MS m/z 571 (M+1).
Anal. Calcd for C30H42N405Sl-1.75 TFA: C, 52.24; H, 5.72, N, 7.27.
Found: C, 52.19; H, 5.82; N, 7.61.
EXAMPLE 29
Preparation of N-[2(S)-(N'-(4-nitrobenzyl)-pyroglutamyl)-amino)-3(S)-
methylpentyl]-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
hvdrochloride salt()
Step A: Preparation of (S)-N-(4-nitrobenzyl)pyroglutamic acid
methyl ester
(S)-Pyro~ t~mic acid methyl ester (0.200 g, 1.40 mmol)
was dissolved in dry THF (5 ml) and NaH (0.061 g, 1.5 mmol) was
added. After gas evolution ceased, 4-nitrobenzyl bromide (0.332 g, 1.54
mmol) was added and the mixture stirred for 1 h. The reaction was
quenched with saturated NaHCO3 solution (40 mL) and extracted with
EtOAc (2 x 50 ml). The organic layers were washed with water, brine,
dried (MgSO4), filtered, and concentrated to give the title compoundas a
solid. 1H NMR (CDC13) ~ 8.19 (d, 2H, J=8.6 Hz), 7.40 (d, 2H, J=8.6
Hz), 5.29 (d, lH, J=15 Hz), 4.19 (d, lH, J=15 Hz), 4.02 (dd, lH, J=3,9
Hz), 3.79 (s, 3H), 2.54-2.67 (m, lH), 2.42-2.51 (m, lH), 2.27-2.39 (m,
lH), 2.11-2.21 (m, lH).
Step B: Preparation of (S)-N-(4-nitrobenzvl)pyro~lutamic acid
(S)-N-(4-Nitrobenzyl)pyroglutamic acid methyl ester (0.365
g, 1.31 mmol) was dissolved in 10 ml MeOH, cooled to 0C, and lN
NaOH (5.2 ml, 5.2 mmol) was added. The reaction was stirred at room
temperature for lh. Water (50 ml) was added and the aqueous was
washed with 2 x 50 ml EtOAc. The aqueous was acidified with 1 N HCl
and extracted with 3 x 40 ml EtOAc. The organic layers were dried
(MgSO4), filtered, and concentrated to give the title compound as a solid.
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lH NMR (d6-DMSO) ~ 8.19 (d, 2H, J=8.7 Hz), 7.51 (d, 2H, J=8.6 Hz),
4.86 (d, lH, J=16 Hz), 4.19 (d, lH, J=16 Hz), 4.02-4.10 (m, lH), 3.30 (br
s, lH), 2.29-2.41 (m, 3H), 1.96-2.05 (m, lH).
Step C: Preparation of N-[2(S)-((4-
Nitrobenzyl)pyroglutamyl)amino)-3(S)-methylpentyl] -N-(1 -
naphthylmethyl)-glycyl-methionine methyl ester
hydrochloride salt
(S)-N-(4-Nitrobenzyl)pyroglutamic acid (0.95 g, 0.36
l0 mmol), N-[2(S)-amino-3-methylpentyl)-N-(1-naphthylmethyl)-glycyl-
methionine methyl ester hydrochloride (10, 0.160 g, 0.300 mmol) and
diisopropylethyl~mine (0.261 mL, 1.50 mmol) were dissolved in DMF (3
mL). BOP-Cl (0.137 g, 0.539 mmol) was added and the mixture was
stirred at ambient temperature for 24 h. The mixture was concelltrated
15 and the residue was partitioned between EtOAc (80 mL) and saturated
NaHCO3 solution (25 mL). The aqueous layer was extracted with
EtOAc (30 mL). The combined organic layer was washed with brine (25
mL), dried (MgSO4), filtered, and concentrated to give a crude product
which was purified by chromatography (silica gel, eluting with 98:2
20 CH2C12:MeOH). FurtherpurificationbypreparativeHPLC(WatersC-
18 Prep Pak eluting with acetonitrile/0.1 % TFA in H2O gradient) gave
the amine trifluoroacetate, which was converted to the hydrochloride salt
by dissolving in EtOAc, bubbling HCl gas, filtering, and drying under
vacuum to give the title compound. 1H NMR (CD30D) o 8.29-8.41 (m,
25 lH), 8.17 (d, 2H, J=8 Hz), 7.92-8.08 (m, 2H), 7.64-7.76 (m, 2H~, 7.48-
7.64 (m, 2H), 7.33-7.48 (m, 2H), 5.03-5.18 (m, lH), 4.59-4.72 (m, lH),
4.39-4.52 (m, lH), 3.81-4.27 (m, 4H), 3.72 (s, 3H),3.14-3.28 (m, lH),
2.50-2.73 (m, lH), 2.19-2.50 (m, 6H), 1.85-2.13 (m, 4H), 2.01 (s, 3H),
1.67-1.85 (m, lH), 1.41-1.53 (m, lH), 1.24-1.38 (m, lH), 1.02-1.19 (m,
30 lH), 0.72-0.94 (m, 6H).
Anal. Calcd for C37H46N507S-1.95 HCl-0.95 H20: C, 56.04; lH, 6.34;
N, 8.83.
Found: C, 56.07; H, 6.28; N, 8.71.
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EXAMPLE 30
Preparation of N-[2(S)-((4-nitrobenzyl)pyroglutamyl)-amino)-3(S)-
methylpentyl]-N-(l-naphthylmethyl)-glycyl-methionine trifluoroacetate
5 salt
N-[2(S)-((4-Nitrobenzyl)pyroglutamyl)amino)-3(S)-
methylpentyl] -N-(1 -naphthylmethyl)-glycyl-methionine methyl ester
(0.050 g, 0.071 mmol) was dissolved in MeOH (1 ml), cooled to 0, and
lN NaOH (0.283 ml, 0.283 mmol) was added. The mixture was stirred at
o ambient temperature for 1 h. The mixture was neutralized with l N HCI
(0.283 ml, 0.283 mmol). The aqueou,s layer was washed with EtOAc
(3x10 ml). The organic layers were combined, dried with MgSO4,
filtered, and concentrated to give a crude product. Preparative HPLC
(Waters C- 1 P~ Prep Pak eluting with acetonitrile/0.1 % TFA in H2O
15 gradient) gave the pure title compound. lH NMR (CD30D); o 8.35 (d,
lH, J=8 Hz), 8.17 (d, 2H, J=8 Hz), 7.94-8.04 (m, 2H), 7.70-7.77 (m, lH),
7.61 (t, lH, J=8 Hz), 7.52-7.63 (m, 2H), 7.42 (d, 2H, J=8 Hz), 4.93-5.10
(m, lH), 4.62-4.75 (m, lH), 4.43-4.56 (m, lH), 4.08-4.21 (m, lH), 3.81-
4.21 (m, 4H), 3.45-3.61 (m, lH), 3.10-3.26 (m, 2H), 2.28-2.53 (m, 6H),
1-95-2.19 (m, 3H), 2.03 (s, 3H) 1.76-1.92 (m, lH), 1.41-1.54 (m, lH),
1.24-1.38 (m, lH), 1.03-1.17 (m, lH), 0.77-0.94 (m, 6H).
Anal. Calcd for C36H44N5O7S-1.9 TFA-0.85 H20: C, 51.80; H, 5.20; N,
7.59.
Found: C, 51.81; H, 5.36; N, 7.53.
EXAMPLE 31
Preparation of N-[2(S)-((N'-benzylpyroglutamyl)amino)-3(S)-
methylpentyl]-N-(1-naphthylmethyl)-glycyl-methionine methyl ester
30 trifluoroacetate salt
Using the method of Example 29, subsliLu~ g benzyl
bromide for the p-nitrobenzyl bromide used therein, the title compound
was obtained.
Anal. Calcd for C37H4gN4O5S-1.65 TFA: C, 57.01; H, 5.89; N, 6.60.
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Found: C, 56.96; H, 5.94; N, 6.91.
EXAMPLE 32
Preparation of N-[2(S)-(N'-benzylpyro-glutamyl)amino)-3(S)-
methylpentyl]-N-(1-naphthylmethyl)-glycyl-methionine trifluoroacetate
salt
The product of Example 31 was converted to the title
compound as described in Example 30.
FAB MS calcd for C36H47N405S 647 (MH+), found 647
Anal. Calcd for C36H46N405S-1.5 TFA: C, 57.27; H, 5.85; N, 6.85.
Found: C, 57.17; H, 5.94; N, 6.79.
EXAMPLE 33
Preparation of N-[2(S)-1 -(4-Fluorophenylmethyl)-1 H-imidazol-5-
ylacetyl)amino-3(S)-methylpentyl]-N- 1 -naphthylmethyl-glycyl-
methionine
Step A: Preparation of 1 -(4-Fluorophenylmethyl)- 1 H-imidazol-5-
ylacetic acid
The title compound was prepared as the hydrogen bromide
salt using the procedures described in Fx~mple 3 steps B and C replacing
4-nitrobenzyl bromide with 4-fluorobenzyl bromide.
lH NMR(CD30D, 400 MHz) ~ 8.89(1H, d, J=1.3Hz), 7.55(1H, s), 7.50-
7.30(2H, m), 7.17(2H, t, J=8.8Hz), 5.43(2H, s) and 3.82(2H, s) ppm.
Step B: Preparation of N-[2(S)-1-(4-Fluorophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N-1-
naphthylmethyl-glycyl-methionine methyl ester bis
trifluoroacetate
The title compound was prepared as the bis trifluoroacetate
salt using the procedures described in example 2 step C using 1-(4-
Fluorophenylmethyl)- 1 H-imidazol-5-ylacetic acid.
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lH NMR(CD30D, 400 MHz) ~ 8.77(1H, s), 8.28(1H,m), 8.00-
7.80(2H,m)), 7.65-7.40(5H,m), 7.30-7.20(2H,m), 7.14(2H,t,
J=8.6Hz),5.34(2H, m) 4.39(2H,m), 4.13(1H,m), 3.68(3H,s), 3.65-
3.40(4H,m), 2.95(1H,m), 2.40-2.15(2H,m), 1.97(3H,s), 1.95(1H,m),
1.70(1H,m), 1.60(1H,m), 1.43(1H,m), 1.07(1H,m), and 1.00-0.80(6H,m)
ppm.
FAB Mass spectrum, m/z = 676 (M+1~.
Anal. calc'd for C37H46NsO4S 0.45H20, 1.65TFA; C, 55.50 H, 5.61 N,
8.03. Found: C, 55.50; H, 5.60; N, 8.23.
Step C: Preparation of N-[2(S)-1-(4-Fluorophenylmethyl)-lH-
imidazol-5-ylacetyl)amino-3(S)-methylpentyl]-N- 1 -
naphthylmethyl-~lycyl-methionine bis trifluoroacetate.
The title compound was prepared as the bis trifluoroacetate
salt using the procedure described in Example 2 step D.
lH NMR(CD30D, 400 MHz) ~ 8.79(1H, s), 8.30(1H,m), 8.00-
7.80(2H,m)), 7.65-7.40(5H,m), 7.30-7.20(2H,m), 7.13(2H,t,
J=8.7Hz),5.35(2H,m) 4.38(2H,m),4.13(1H,m),3.80-3.40(4H,m),
3.10(1H,m), 2.40-2.15(2H,m), 1.97(3H,s), 1.95(1H,m), 1.70(1H,m),
1.60(1H,m), 1.43(1H,m), 1.07(1H,m), and 1.00-0.80(6H,m) ppm.
FAB Mass spectrum, m/z = 662 (M+1).
Anal. calc'd for C36H44N5O4S 0.60H20, 2.30TFA; C, 52.16 H, 5.12 N,
7.49. Found: C, 52.18; H, 5.13; N, 7.76.
EXAMPLE 34
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(l -naphthylmethyl)glycyl-
methionine isopropyl ester
Step A: Preparation of lH-Imidazole-4- acetic acid methyl ester
hydrochloride.
A solution of lH-imidazole-4-acetic acid hydrochloride
(4.00g, 24.6 mmol) in methanol (100 ml) was saturated with gaseous
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hydrogen chloride. The resulting solution was allowed to stand at room
temperature (RT) for 18hr. The solvent was evaporated in vacuo to afford
the title compound as a white solid.
lH NMR(CDC13, 400 MHz) ~ 8.85(1H, s),7.45(1H, s), 3.89(2H, s) and
3.75(3H, s) ppm.
Step B: Preparation of 1 -(Triphenylmethyl)-lH-imidazol-4-ylacetic
acid methvl ester.
To a solution of the product from Step A (24.85g, 0.141mol)
in dimethyl formamide (DMF) (115ml) was added triethylamine (57.2
ml, 0.412mol) and triphenylmethyl bromide(55.3g, 0.171mol) and the
suspension was stirred for 24hr. After this time, the reaction mixture was
diluted with ethyl acetate (EtOAc) (1 1) and water (350 ml). The organic
phase was washed with sat. aq. NaHCO3 (350 ml), dried (Na2S04) and
evaporated in vacuo. The residue was purified by flash chromatography
(sio2~ 0-100% ethyl acetate in hexanes; gradient elution) to provide the
title compound as a white solid.
1H NMR (CDCl3, 400 MHz) ~ 7.35(1H, s), 7.31(9H, m), 7.22(~H, m),
6.76(1H, s), 3.68(3H, s) and 3.60(2H, s) ppm.
Step C: Preparation of [1-(4-cyanobenzyl)-lH-imidazol-S-yl]acetic
acid methyl ester.
To a solution of the product from Step B (8.00g, 20.9mmol)
in acetonitrile (70 ml) was added bromo-p-toluonitrile (4.10g, 20.92
mmol) and heated at 55C for 3 hr. After this time, the reaction was
cooled to room temperature and the resulting imidazolium salt (white
precipitate) was collected by filtration. The filtrate was heated at 55C
for 18hr. The reaction mixture was cooled to room temperature and
evaporated in vacuo. To the residue was added EtOAc (70 ml) and the
resulting white precipitate collected by filtration. The precipitated
imidazolium salts were combined, suspended in methanol (100 ml) and
heated to reflux for 30min. After this time, the solvent was removed in
vacuo, the resulting residue was suspended in EtOAc (75ml) and the solid
isolated by filtration and washed (EtOAc). The solid was treated with sat
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aq NaHC03 (300ml) and CH2C12 (300ml) and stirred at room
t temperature for 2 hr. The organic layer was separated, dried (MgS04)
and evaporated in vacuo to afford the title compound as a white solid:
lHNMR(CDC13, 400 MHz) o 7.65(1H, d, J=8Hz), 7.53(1H, s), 7.15(1H,
d, J=8Hz), 7.04(1H, s), 5.24(2H, s), 3.62(3H, s) and 3.45(2H, s) ppm.
Step D: Preparation of [1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetic
acid.
A solution of [1-(4-cyanobenzyl)-lH-imidazol-5-yl]acetic
o acid methyl ester (4.44g, 17.4mmol ) in THF (lOOml) and 1 M lithium
hydroxide (17.4 ml, 17.4 mmol) was stirred at RT for 18 hr. 1 M HCl
(17.4 ml) was added and the THF was removed by evaporation in vacuo.
The aqueous solution was lyophilised to afford the title compound
cont~inin~ lithium chloride as a white solid.
H NMR(CD30D, 400 MHz) d 8.22(1H, s), 7.74(1H, d, J=8.4Hz),
7.36(1H, d, J=8.4Hz), 7.15(1H, s), 5.43(2H, s) and 3.49(2H, s) ppm.
Step E: Preparation of N-[2(S)-(amino)-3(S)-methylpentyl]-N-( l -
naphthylmethyl)~lycine methyl e~ter hydrochloride.
A solution of N-[2(S)-(t-Butoxycarbonylamino)-3(S)-
methylpentyl]N(l-naphthylmethyl) glycine methyl ester from example 1
step E (5.90g, 13.8 mmol) in EtOAc (lOOml) was saturated with gaseous
hydrogen chloride. The resulting solution was allowed to stand at room
temperature for lhr. The solvent was evaporated in vacuo to afford the
2s title compound as a white solid.
lH NMR(CD30D 400 MHz) ~ 8.26(1H, d, J=8.6Hz),7.92(1H, d,
J=7.2Hz), 7.87(1H, d, J=8.6Hz), 7.63-7.42(4H,m), 4.34(1H,d, J=12.3Hz),
4.26(1H,d, J=12.3Hz), 3.68(3H,s), 3.13(1H, d, J=10.3Hz), 2.67-
2.55(2H,m), 1.46(1H,m), 1.28(2H,m), 1.10-0.90(2H,m),
0.84(3H,d,J=6.8Hz) and 0.77(3H,t, J=6.8Hz)ppm.
Step F: Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-
5- yl] acetylamino)-3(S)-methylpentyl]-N-(1-
naphthylmethyl) ~lvcine methvl ester.
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To a solution of [1-(4-cyanobenzyl)-lH-imidazol-5~yl]acetic
acid. (4.09g, 10.24 mmol), the amine hydrochloride salt from step
E(5.07g, 10.24 mmol), HOOBT (1.67g, 10.24mmol), and N-
methylmorpholine (2.36ml, 21.5mmol) in DMF (SOml) at 0C, was added
EDC (2.16g, 11.26 mmol). The reaction was stirred at room temperature
for l 8hrs, diluted with EtOAc and the organic layer washed with sat. aq
NaHCO3, brine, dried (Na2SO4), and the solvent evaporated in vacuo.
The residue was chromatographed (sio27 3-4% MeOH in CH2C12) to
afford the title compound as a white solid.
1H NMR(CD30D, 400 MHz) ~ 8.30(1H,d, J=8.4Hz),
7.84(1H,d, J=8.0Hz), 7.80(1H,t, J=4.5Hz), 7.68-7.38(3H,m), 7.48-
7.32(4H,m), 7.10(2H,d, J=8.0Hz), 6.87(1H,s), 5.24(1H,d, J=16.7Hz),
5.18(1H,d,J=16.7Hz), 4.83(2H,s), 4.27(1H,d, J=12.8Hz), 4.10(1H,d,
J=12.8Hz), 3.97(1H,m), 3.65(3H,s), 3.40-3.20(2H,m), 2.92(1H,dd,
J=13.3 and 4.3Hz), 2.60(1H,dd, J=13.3 and 10.0Hz), 1.48(1H,m),
1.25(1H,m), 0.98(1H,m), 0.78(3H,d, J=6.8Hz) and 0.77(3H,t, J=7.5Hz)
ppm.
Anal. calc'd for C33H37N503 l.05H20, 2.85 TFA C, 51.90; H, 4.72; N,
7-82- Found: C, 51.90; H, 4.70; N, 8.18.
FAB Mass spectrum, m/z = 552 (M+1).
Step G: Preparation of N-[2~S)-([1-(4-cyanobenzyl)-lH-imidazol-
5-yl]acetylamino)-3(S)-methylpentyl] -N-(1 -naphthylmethyl)
glycine.
A solution of the methyl ester from step F (2.32g,
4.21mmol) in MeOH(20ml) and 1 M lithium hydroxide (4.70 ml, 4.70
mmol) was stirred at RT for 6hr. The aqueous solution diluted with water
(lSml) and extracted with EtOAc (lOOml), dried (Mg2SO4), and the
3 0 solvent evaporated in vacuo. The residue was chromatographed (siO2,
20% MeOH in CH2Cl2) to afford the title compound as a white solid.
1H NMR(CD30D, 400 MHz) ~ 8.33(1H, d,J=8.3Hz),
7.87(2H,d, J=7.7Hz), 7.78(1H,s), 7.63(2H,d, J=6.6Hz), 7.57(1H,dl,
J=6.4Hz), 7.50-7.38(4H,m), 7.17(1H,d, J=8.3Hz), 6.96(1H,s), 5.32(1H,d,
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J=16.6Hz),5.25(1H,d,J=16.6Hz),4.64(1H,d, J=13.2Hz),4.40(1H,d,
J=13.2Hz),3.99(1H,m),3.60-3.28(4H,m),3.22(1H,dd, J=13.3 and
3.1Hz), 2.93(1H,dd, J=13.3 and 10.3Hz), 1.52(1H,m), 1.29(1H,m),
1.06(1H,m), 0.86-0.76(6H,m) ppm.
Anal. calc'd for C32H35N5O3 l.00H20, C,69.17; H,6.71 N,12.60.
Found: C,68.95; H,6.37; N,12.54.
FAB Mass spectrum, m/z = 538 (M+l).
Step H: Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-
o imidazol-5-yl]acetylamino)-3(S)-methylpentyl]-N-( l -
naphthvlmethyl)~lycvl-methionine isopropyl ester
To a solution of the acid from step G (lOOmg, 0.186mmol)
and methionine isopropyl ester hydrochloride (42.4mg,0.186mmol),
HOOBT (30.4mg, 0.186mmol) and triethylamine (0.077ml, 0.56mmol)
in DMF (l.Oml) was added EDC (37.5mg, 1.96mmol). The reaction was
stirred at room temperature for 18hrs, diluted with EtOAc and the organic
layer washed with sat. aq NaHCO3, brine, dried (Na2SO4), and the
solvent evaporated in vacuo. The residue was chromatographed (sio2
20 5% MeOH in CH2Cl2), evaporated to dryness and converted to the
hydrochloride salt by treatment with aqueous HCl (0.32ml of a 1 M
solution) and acetonitrile and lyophili~tion, to afford the title compound
as a white powder.
lH NMR(CD30D,400 MHz) ~ 9.00-8.90(1H, m), 8.38(1H, m),8.10-
25 7.10(1 lH,m),5.80-4.80 (4H, m),4.60-3.30(1 lH,m), 2.60-1.70(8H,m),
1.60(1H,m),1.42(1H,m),1.21(6H,d,J=6.2Hz),0.918(6H,brt,J=7.3Hz)
ppm.
FAB HRMS exact mass calc'd for C40H5lN6o4s 711.369251 (MH+),
found 711367663.
30 Anal. calc'd for C40H50N6o4s 0.55H20 and 2.80HCl C,58.38; H, 6.60
N,10.21. Found: C,58.40; H, 6.60; N,10.36.
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EXAMPLE 35
Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-
imidazol-5-yl]acetylamino)-3(S)-methylpentyl]-N-(1-
5 naphthvlmethyl)~lvcvl-methionirle sulfone methyl ester
The title compound was prepared as the hydrogen chloride
salt using the procedures described in Example 34 Steps H using
methionine sulfone methyl ester hydrochloride.
o lH NMR(CD30D, 400 MHz) ~ 8.93(1H, m), 8.39(1H, m),8.20-
7.15(1 lH,m),5.50(2H,m), 5.40-3.00 (lSH, m), 2.95(3H,s), 2.30(1H,m),
2.05(1H,m), 1.60(1H,m), 1.45(1H,m), 1.22(1H,m), 0.915(6H,m) ppm.
PAB HRMS exact mass calc'd for C3gH47N6O6S 715.327781(MH+),
found 715.327372.
Anal. calc'd for C38H47N606S 0.35H20 and 3.25HCl C, 54.36; H, 6.00
N, 10.01. Found: C, 54.36; H, 5.99; N, 10.21.
EXAMPLE 36
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- lH-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-
methionine sulfone
A stirred solution of the methyl ester from Example 35
(23.7mg, 0.033mmol) in TH~(0.20ml) and 1 M lithium hydroxide
(0.033ml, 0.033mmol)was allowed to warm from 0C to room
temperature over 18hrs. The reaction was quenched by the addition of
trifluoroacetic acid and the solvent evaporated in vacuo. The residue was
30 purified by preparative hplc to afford the title compound after
lyophili~tion.
1H NMR(CD30D, 400 MHz) ~ 8.89(1H, m), 8.16(1H, m), 7.8S-
7.20(11H,m),5.38(2H,m), 4.31(1H,m), 4.00(1H,m), 3.60-3.30(7H,m),
3.00-2.90(3H,m), 2.81(3H,s), 2.14(1H,m), 1.94(1H,m), 1.431H,m),
1.29(1H,m), 1.04(1H,m), 0.78(6H,m) ppm.
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- Anal. calc'd for C37H44N606S 0.45H20, 2.30 TFA C, 51.45; H, 4.90 N,
8.65. Found: C, 51.44 H, 4.89; N, 8.62.
FABHRMS exactmasscalc'dfor C37H45N6O6S 701.312130(MH+),
found 701.313179.
EXAMPLE 37
Preparation of N-[2(S)-([ 1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-(3-
o acetylamino)alanine methyl ester
The title compound was prepared as the hydrochloride saltusing the procedures described in Example 34 Step H using (S)-N'-acetyl
minopropionic acid methylester hydrochloride.
H NMR(CD30D, 400 MHz) ~ 8.90(1H, m), 8.38(1H, m),8.10-
7.20(1 lH,m), 5.60(2H,m), 5.20-3.00(10H,m), 3.60(3H,s), 1.92(3H,s),
1.83(1H,s), 1.57(1H,m), 1.43(1H,m), l.l9(1H,m), 0.90(6H,m) ppm.
FAB HRMS exact mass calc'd for C3gH46N7O5 680.356043(MH+),
found 680.356735.
20 Anal. calc'd for C3gH45N7O5 0.35H20 and 3.05 HCl C, 57.24; H, 6.16
N, 12.30. Found: C, 57.26; H, 6.16; N, 12.40.
EXAMPLE 38
25 Preparation of N-[2(S)-([1 -(4-cyanobenzyl)-lH-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1-naphthylmethyl)glycyl-(3-
acetylamino)alanine
3 0 The title compound was prepared as the trifluoroacetate salt
using the procedures described in Example 36 and the methyl ester
prepared in Example 37
1H NMR(CD30D, 400 MHz) ~ 8.82(1H, m), 8.40(1H, m), 7.70(2H,m),
7.65(2H,d, J=8.0Hz), 7.60-7.30(5H,m), 7.27(2H,d, J=8.0Hz),
5.40(2H,m),
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4.32(1H,m), 4.00(1H,m), 3.70-3.10(10H,m), 1.75(3H,s), 1.48(1H,s),
1.33(1H,m), 1.08(1H,m), 0.80(6H,m) ppm.
FAB HRMS exact mass calc'd for C37H44N705 666.340393(MH+),
found 666.340627.
Anal. calc'd for C37H43N705 0.30H20 and2.35 TFA C, 53.33; H, 4.93
N, 10.44. Found: C, 53.33; H, 4.95; N, 10.22.
EXAMPLE 39
o Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-2(RS)
amino-3-(2 thienyl)propionic acid methyl ester
The title compound was prepared as the trifluoroacetate salt
using the procedures described in Example 34 Step H using 2(RS) amino-
3-(2 thienyl)propionic acid methyl ester hydrochloride.
lH NMR(CD30D, 400 MHz) ~ 8.81(1H, m), 8.19(1H, d, J=9.OHz),
8.00-7.80(2H,m), 7.62(2H,d, J=8.0Hz), 7.50-7.30(5H,m), 7.29(2H,d,
J=8.0Hz), 7.036(1H,m), 6.718(1H,s), 6.61(1H,m), 5.39(2H,m),
4.60(1H,m), 4.40(1H,m), 3.98(1H,m), 3.60(3H,s), 3.60-3.30(7H,m), 3.20-
2.95(3H,m), 1.47(1H,m), 1.32(1H,m), 1.08(1H,m), 0.85(6H,m) ppm.
FAB HRMS exact mass calc'd for C40H45N604S 705.322301 (MH,
found 705.321444.
Anal. calc'd for C40H44N604S 0.35H20 and 2.50TFA C, 54.25; H,
4.78 N, 8.44. Found: C, 54.27; H, 4.77; N, 8.36.
EXAMPLE 40
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylrnethyl)glycyl-2(RS)-
amino-3-(2 thienyl)propionic acid
The title compound was prepared as the trifluoroacetate salt using the
procedures de~cribed in Example 36 and the methyl ester prepared in
Example 39
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FAB HRMS exact mass calc'd for C39H42N6O4S 691.306651 (MH+),
found 691.306950.
EXAMPLE 41
5 Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-2(S)
amino-4-sulfamyl-butanoic acid methyl ester
The title compound was prepared as the trifluoroacetate salt
using the procedures described in Example 34 Step H using 2(S) amino-
4-sulfamyl-butanoic acid methyl ester hydrochloride.
1H NMR(CD30D, 400 MHz) o 8.87(1H, m), 8.33(1H, m), 8.00-
7.80(2H,m), 7.73(2H,d, J=8.2Hz), 7.70-7.40(5H,m), 7.35(2H,d,
J=8.0Hz),
15 5.42(2H,m), 4.40(1H,m), 4.10(1H,m), 3.70(3H,s), 3.60-3.20(7H,m),
3.00(3H,m),2.30(1H,m), 2.05(1H,m), 1.55(1H,m), 1.40(1H,m),
1.15(1H,m), 0.95(6H,m) ppm.
FAB HRMS exact mass calc'd for C37H46N7O6S 716.323030(MH+),
found 716.323766.
Anal. calc'd for C37H4sN7O6S 1.20H20 and 3.00TFA C, 47.84; H,
4.71 N, 9.08. Found: C, 47.84; H, 4.58; N, 9.26.
EXAMPLE 42
Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-2(S)
amino-4-sulfamvl-butanoic acid
The title compound was prepared as the trifluoroacetate salt
using the methyl ester prepared in Example 41.
30 1H NMR(CD30D, 400 MHz) ~ 8.86(1H, m), 8.26(1H, m), 8.00-
7.80(2H,m), 7.73(2H,d, J=8.2Hz), 7.70-7.40(5H,m), 7.35(2H,d,
J=8.0Hz),
~- .
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5.47(2H,m), 4.42(1H,m), 4.08(1H,m), 3.60-3.20(7H,m), 3.00(3H,m),
2.30(1H,m), 2.05(1H,m), 1.57(1H,m), 1.38(1H,m), l.lS(lH,m),
0.95(6H,m) ppm.
FAB HRMS exact mass calc'd for C36H44N7O6S 702.307379(MH+),
found 702.308307.
Anal.calc'dforC36H43N7O6S 0.40H20and2.65TFA C,49.06;H,
4.63 N, 9.70. Found: C, 49.03; H, 4.63; N, 9.99.
EXAMPLE 43
Preparation of N-r2(S)-([1-(4-cyanobenzyl)-lH-imidazol-S-
yl]acetylamino)-3(S)-methylpentyl]-N-~l -naphthylmethyl)glycyl-N-
methyl methionine methyl ester
The title compound was prepared as the trifluroacetate salt
using the procedures described in Example 34 Step H using N-methyl
me~ionine methyl ester hydrochloride.
1H NMR(CD30D, 400 MHz) ~ 8.93(1H, m), 8.34(1H, m), 8.04(1H,d,
J=7.7Hz), 7.98(1H,m), 7.75(3H,m), 7.60-7.20(6H,m), 5.48(2H,m),
5.06(1H,m), 4.40(1 H,m), 4.10(1H,m), 3.66~3H,s), 3.80-3.20(9H,m),
2.85(3H,br s), 2.40-2.00(1H,m),2.05(3H,s), 1.95(lH,m), 1.57(1 H,m),
1.45(1H,m), l.lO(lH,m), 0.95(6H,m) ppm.
FAB HRMS exact mass calc'd for C39H49N604S 697.353601 (MH+),
found 697.353335.
Anal. calc'd for C39H4gN6O4S 0.45H20 and 2.95TFA C, 51.79, H,
5.02 N, 8.07. Found: C, 51.79; H, 4.99; N, 8.15.
EXAMPLE 44
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-S -
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-N-
methyl methion~e
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The title compound was prepared as the trifluoroacetate salt
using the procedures described in Example 36 and the methyl ester
prepared in Example 43.
lH NMR(CD30D, 400 MHz) ~ 8.78(0.7H, m),8.76(0.3H,m), 8.24(1H,
m),8.0-7.00(11H,m), 5.37(2H,m), 5.00-3.00(10H,m),2.85(3H,brs),
2.40-2.00(4H,m),1.93(0.9H,s), 1.90(2.1H,m), l.50(1H,m), 1.31(1H,m),
1.08(1H,m), 0.80(6H,m) ppm.
FAB HRMS exact mass calc'd for C36H47N6O4S 683.337951 (MH+),
found 683.337329.
o Anal. calc'd for C36H46N6O4S 2.84TFA C, 52.11; H, 4.89 N, 8.35.
Found: C, 51.74; H, 5.02; N, 8.74.
EXAMPLE 45
Preparation of N-[2(S)-(~1-(4-cyanobenzyl)-lH-imidazol-5-
yl] acetylamino)-3 (S)-methylpentyl] -N-( l -naphthylmethyl)glycyl -
homoserine lactone
The title compound was prepared as the trifluoroacetate salt
20 using the procedures described in Example 34 Step H using homoserine
lactone hydrochloride.
lH NMR(CD30D, 400 MHz) ~ 8.91(1H, m), ~s.30(1H, m), 8.05-
7.90(2H, m), 7.74(2H,d, J=8.4Hz),7.70(1H,d, J=6.2Hz), 7.60-
7.50(4H,m), 7.53(2H,d, 8.0Hz), 5.50(2H,m), 4.70(2H,m),
2s 4.39(1H,dd,J=10.9 and 8.9 Hz), 4.30(1H,t, J=7.9Hz), 4.21(1H,m),
4.05(2H,m), 4.00-3.40(5H,m), 2.30(1H,m), l.90(1H,m), 1.57(1H,m),
1.43(1H,m), 1.18(1H,m), 0.98-0.90(6H,m) ppm.
FAB HRMS exact mass calc'd for C36H4lN6o4 621.318929(MH+),
found 621.317455.
Anal. calc'd for C39H4gN6O4S 0.83H20 and 3.76TFA C, 49.11; H,
3 4.30 N, 7.90. Found: C, 49.11; H, 4.30; N, 8.35.
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EXAMPLE 46
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl] ace~.ylamino)-3 (S )-methylpentyl] -N-(1 -naphthylmethyl)glycyl -
5 homoserine
The title compound was prepared as the lithium salt usingthe procedures described in Example 36 and the lactone prepared in
Example 45.
FABHRMS exactmasscalc'dforC36H43N60s 639.329494(MH+),
found 639.328919.
EXAMPLE 47
15 Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(l -naphthylmethyl)glycyl-proline
methyl ester
The title compound was prepared as the trifluoroacetate salt
20 using the procedures described in Example 34 Step H using L- proline
methyl ester hydrochloride.
lH NMR(CD30D, 400 MHz) ~ 8.80(1H~ s), 8.38-8.28(1H,m),
8.02(1H, d, J=8.4Hz), 7.96(1H, d, J=8.4Hz), 7.80-7.65(3H,m), 7.60-
7.30(6H,m), 5.55-5.40(2H,m), 5.00(1H,m), 4.40-4.00(3H,m),3.70(3H,m),
25 3.70-3.00(8H,m), 2.25-2.05(1H,m), 2.00(2H,m), 1.95-1.50(2H,rn),
1.40(1H,m), 1.17(1H,m), 1.00-0.80(6H,m)ppm.
FAB HRMS exact mass calc'd for C3gH45N6O4 649.350229(MH+),
found 649.350481.
Anal. calctd for C3gH44N6O4 1.75H20 and 3.00TFA C, 51.69; H,
4.98N, 8.22. Found: C, 51.69; H, 4.79; N, 8.58.
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EXAMPLE 48
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl] acetylamino)-3 (S )-methylpentyl] -N-(1 -naphthylmethyl)glycyl-
5 prOllIle
The title compound was prepared as the trifluoroacetate saltusing the procedures described in Example 36 and the methyl ester
prepared in Example 47.
o lH NMR(CD30D, 400 MHz) o 8.85(0.8H, m), 8.80(0.2H,m), 8.32(1H,
d, J=8.4Hz), 8.04-7.90(2H,m), 7.80-7.64(3H,m), 7.60-7.28(6H,m),5.54-
5.36(2H,m), 4.40-4.00(2H,m), 3.85-3.00(10H,m), 2.20(1H,m), 2.10-
1.80(3H,m), 1.57(1H,m), 1.42(1H,m), 1.17(1H,m), 0.98-0.82(6H,m)
ppm.
15 FABHRMS exactmasscalc'dforC37H43N604 635.334579(MH+),
found 635.332994.
Anal. calc'd for C37H42N604 0.80H20 and 2.80TFA C, 52.83; H,
4.83N, 8.68. Found: C, 52.81, H, 4.81; N, 8.88.
EXAMPLE 49
Preparation of N-[2(S)-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-D-
25 proline methyl ester
The title compound was prepared as ~e trifluoroacetate saltusing the procedures described in Example 34 Step H using D- proline
methyl ester hydrochloride.
lH NMR(CD30D, 400 MHz) ~ 8.92(0.3H, s), 8.88(0.7H,s),8.08-
7.90(2H,m), 7.85-7.30(10H,m), 5.46(2H,m), 5.00-4.40(1H,m),
4.35(1H,m), 4.10-4.00(2H,m), 3.60(3H,s), 3.80-3.20(8H,m), 2.20(1H,m),
2.00-1.80(3H,m), 1.60(1H,m), 1.45(1H,m), 1.15(1H,m), 1.00-
0.80(6H,m)ppm.
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FAB HRMS exact mass calc'd for C3gH45N6O4 649.350229(MH+),
found 649.351271.
Anal. calc'd for C3gH44N6O4 2.20H20 and 3.00TFA C, 51.28, H,
5.03N, 8.16. Found: C, 51.27, H, 4.71 N, 8.39.
EXAMPLE 50
Preparation of N- [2(S )-([1 -(4-cyanobenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl] -N-(1 -naphthylmethyl)glycyl-
proline
The title compound was prepared as the tri~luoroacetate salt
using the procedures described in Example 36 and ~e methyl ester
prepared in Example 49.
H NMR(CD30D, 400 MHz) ~ 8.80-8.70(1H, m), 8.30-8.15(1H, m),
8.00-7.20(1 lH,m), 5.40(0.4H,s), 5.35(1.6H,m), 5.00-4.60(1H,m),
4.24(1H,m), 3.97(1H,m), 3.70-3.00(10H,m), 2.20-2.00(1H,m), 2.00-
1.60(2H,m), 1.50~1H,m), 1.34(1H,m), 1.08(1H,m), 1.90-0.70(6H,m)ppm.
FAB HRMS exact mass calc'd for C37H43N604 635.334579(MH~),
20 found 635.333794.
Anal. calc'd for C37H42N604 0.50H20 and 2.55TFA C, 54.11 H,
4.91N, g.99. Found: C, 54.11; H, 4.93; N, 8.95.
EXAMPLE 51
Preparation of N-[2(S)-([1-(4-cyanobenzyl)-lH-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-L-
pipecolinic acid
The title compound was prepared as the trifluroacetate salt
30 using the procedures described in Example 34 Step H using L-pipecolinic
acid .
lH NMR(CD30D, 400 MHz) ~ 8.96-8.84(1H,m),8.36(1H,m), 8.10-
7.20(1 lH,m), 5.45(2H,m), 5.20-4.40(1H,m), 4.40-4.00(3H,m), 4~00-
3.00(9H,m), 2.20(2H,m), 1.80-1.05(6H,m), 1.00-0.80(6H,m)ppm.
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FAB HRMS exact mass calc'd for C3gH45N6O4 649.350229(MH+),
found 649.352801.
Anal. calc'd for C3gH44N6O4 2.75TFA C, 54.29, H, 4.90N, 8.73.
Found: C, 54.22; H, 4.88 N, 8.89.
EXAMPLE 52
Preparation of N-[2(S)-([1 -(4-carbomethoxybenzyl)- 1 H-imidazol-5-
yl] acetylamino)-3 (S )-methylpentyl] -N-(1 -naphthylmethyl)glycyl -
methionine methyl ester
The title compound -as the trifluoroacetate salt- was isolated
as a minor component of the reaction mixture prepared in Example 9
Step A .
H NMR(CD30D, 400 MHz) o 8.93(1H,s),8.30(1H,m), 8.05-
7.35(9H,m), 7.31(2H,d, J=8.2Hz), 5.48(2H,m), 5.00-4.40(1H,m),
4.39(1H,s), 4.05(1H,m), 3.90(3H,m), 4.00-3.30(7H,m), 3.67(3H,m),
3.17(1H,m), 2.20-2.10(2H,m), 1.98(3H,s), 1.75(1H,m), 1.55(1H,m),
1.40(1H,m), 1.18(1H,m), 1.00-0.80(6H,m)ppm.
20 Anal.calc'dforC39H49N5O6S 0.15H20,2.15TFA C,53.96;H,5.38;
N, 7.27. Found: C, 53.96; H, 5.39 N, 7.59.
EXAMPLE 53
2 5 Preparation of N-[2(S)-([1 -(4-carbomethoxybenzyl)- 1 H-imidazol-5-
yl]acetylamino)-3(S)-methylpentyl]-N-(1 -naphthylmethyl)glycyl-
methionine
The title compound was prepared as the trifluoroacetate salt
30 using the procedures described in Example 36 and the methyl ester
prepared in Example 52.
lH NMR(CD30D, 400 MHz) ~ 8.80(1H, m), 8.20(1H, m), 8.00-
7.20(1 lH,m), 5.40(2H,m), 5.00-4.60(1 H,m), 4.32(1H,m), 4.05( lH,m),
3.80(3H,s), 3.70-3.00(7H,m), 2.40-2.00(3H,m), 1.88(3H,s), 1.75(1H,m),
1.55(1H,m), 1.30(1H,m), 1.05(1H,m), 1.00-0.65(6H,m)ppm.
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Anal. calc'd for C3gH47N506S 0.15H20 and 2.85TFA C, 50.98 H,
4.91N, 6.80. Found: C, 50.98; H, 4.89; N, 7.19.
EXAMPLE 54
Preparation of 1 -(2-naphthylmethyl)- 1 H-imidazol-5-ylacetyl-
isoleucinyl- phenvlalaninyl-methionine methyl ester
o The title compound was prepared as the trifluoroacetate salt
using the procedures described in Example 34 Step H and isoleucinyl-
phenyl~l~ninyl-methionine methyl ester hydrochloride.
lH NMR(CD30D, 400 MHz) ~ 8.89(1H, s), 8.39(1H,d, J=8.0Hz),
8.19(2H,m), 8.00-7.90(3H,m), 7.67(1H,s), 7.60-7.52(2H,m), 7.48(1H,s),
7.36(1H,d,J=8.0Hz), 7.30-7.10(5H,m), 5.56(1H,d,J=15.0Hz),
5.49(1H,dJ=l5.OHz), 4.69(1H,m), 4.52(1H,m), 4.20-4.14(1H,m),
3.54(1H,d, J=18.0Hz), 3.66(1H,d,J=18.0Hz), 3.66~3H,s),
3.14(1H,dd,J=15.0 and 6.0Hz), 2.91(1H,dd, J=15.0 and 9.0Hz), 2.56-
2.16(2H,m), 2.06(1H,m), 2.04(3H,s), 1.89(1H,m), 1.73(1H,m),
20 1.40(1H,m), 1.08(1H,m), 0.90-0.80(6H,m)ppm.
FAB HRMS exact mass calc'd for C37H46N505S 672.321967(MH+),
found 672.321794.
Anal. calc'd for C37H45N505S O.lOH20 and 2.30TFA C, 57.87; H,
5.70N, 8.52. Found: C, 57.88; H, 5.61 N, 8.49.
EXAMPLE 55
Preparation of 1 -(2-naphthylmethyl)- 1 H-imidazol-5-ylacetyl-isoleucinyl-
phenvlalaninyl-methionine
The title compound was prepared as the trifluoroacetate salt
using the procedures described in Example 36 and the methyl ester
prepared in Example 54.
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1H NMR(CD30D, 400 MHz) ~ 8.80(1H, s), 8.15(1H,d, J=8.0Hz),
7.93(1H,d,J=~.OHz), 7.89(2H,m), 7.74(1H,m), 7.58-7.52(2H,m),
7.44(1H,s),
7.35(1H,dd, J=10.0 and 3Hz), 7.30-7.10(5H,m), ~.54(1H,d,J=15.0Hz),
5.47(1H,d, J=15.0Hz), 4.70(1H,m), 4.50(1H,m), 4.15(1H,m), 3.51(1H,d,
J=17.0Hz), 3.66(1H,d,J=17.0Hz), 3.18(1H,dd,J=15.0 and 6.0Hz),
2.92(1H,dd, J=15.0 and 9.0Hz), 2.56-2.40(2H,m), 2.10(1H,m),
2.05(3H,s), 1.92(1H,m), 1.73(1H,m), 1.40(1H,m), 1.08(1H,m), 0.90-
0.80(6H,m)ppm.
FAB HRMS exact mass calc'd for C36H44N505S 658.305448(MH+),
o found 658.306317.
EXAMPLE 56
In vitro inhibition of ras farne.svl transferase
Assays offarnesyl-protein transferase. Partially purified bovine FPTase
and Ras peptides (Ras-CVLS, Ras-CVIM and RAS-CAIL) were prepared
as described by Schaber et al., J. Biol. Chem. 265:14701-14704 (1990),
Pompliano, et al., Biochemistry 31 :3800 (1992) and Gibbs et al., PNAS
U.S~. 86:6630-6634 (1989), respectively. Bovine FPTase was assayed
in a volume of 100,ul cont~ining 100 mM N-(2-hydroxy ethyl)
piperazine-N'-(2-ethane sulfonic acid) (HEPES), pH 7.4, 5 mM MgCl2, 5
mM dithiothreitol (DTT), 100 mM [3H]-farnesyl diphosphate ([3H]-FPP;
740 CBq/mmol, New England Nuclear), 650 nM Ras-CVLS and 10
2s ~lg/ml FPTase at 31C for 60 min. Reactions were initi~ted with FPTase
and stopped with 1 ml of 1.0 M HCL in ethanol. Precipitates were
collected onto filter-mats using a TomTec Mach II cell harvestor, washed
with 100% ethanol, dried and counted in an LKB ,I~-plate counter. The
r assay was linear with respect to both substrates, FPTase levels and time;
less than 10% of the [3H]-FPP was utilized during the reaction period.
Purified compounds were dissolved in 100% dimethyl sulfoxide (DMSO)
and were diluted 20-fold into the assay. Percentage inhibition is
measured by the amount of incorporation of farnesyl in the presence of
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the test compound when compared to the amount of incorporation in the
absence of the test compound.
Human FPTase was prepared as described by Omer et al.,
Biochemistry 32:5167-5176 (1993). Human FPTase activity was assayed
as described above with the exception that 0.1 % (w/v) polyethylene
glycol 20,000, 10 ,UM ZnCl2 and 100 nM Ras-CVIM were added t~ the
reaction mixture. Reactions were performed for 30 min., stopped; with
100 ,ul of 30% (v/v) trichloroacetic acid (TCA) in ethanol and processed
as described above for the bovine enzyme.
The compounds of the instant invention were tested for
inhibitory activity against hllm~n FPTase by the assay described above
and were found to have ICso of ~ 10 ~lM.
EXAMPLE 57
In vivo ras farnesylation assay
The cell line used in this assay is a v-ras line derived from
either Ratl or NIH3T3 cells, which expressed viral Ha-ras p21. The
assay is performed essentially as described in DeClue, J.E. et al., Cancer
Research 51:712-717, (1991). Cells in 10 cm dishes at 50-75%
confluency are treated with the test compound (final concentratiom of
solvent, methanol or dimethyl sulfoxide, is 0.1%). After 4 hours at 37C,
the cells are labelled in 3 ml methionine-free DMEM supple-meted with
10% regular DMEM, 2% fetal bovine serum and 400
mCi[35S]methionine (1000 Ci/mmol). After an additional 20 hours, the
cells are lysed in 1 ml lysis buffer (1 % NP40/20 mM HEPES, pH 7.5/5
mM MgC12/lmM DTT/10 mg/ml aprotinen/2 mg/ml leupeptin/2 mg/ml
antipain/0.5 mM PMSF) and the lysates cleared by centrifugation at
100,000 x g for 45 min. Aliquots of lysates cont~inin~ equal numbers of
acid-precipitable counts are bought to 1 ml with IP buffer (lysis buffer
lacking DlT) and immunoprecipitated with the ras-specific monoclonal
antibody Y13-259 (Furth, M.E. et al., J. Virol. 43:294-304, (1982)).
Following a 2 hour antibody incubation at 4C, 200 ml of a 25%
suspension of protein A-Sepharose coated with rabbit anti rat IgG is
